professor maria ławrynowicz was born on 17 may 1943 at mstów near częstochowa. between 1961 and 1966 she studied in the faculty of biology and earth sciences, university of łódź and, from 1 october 1965, while still a student, was employed at the chair of plant systematics and geography. professor jakub mowszowicz, head of the chair, gave maria the challenging task of setting up a specialist group in mycology. in parallel with her scientific re­ search and teaching, she started to organize, forming a mycological laboratory and gathering around her students interested in fungi. from the outset she made it her priority to assemble collections of larger fungi and, particularly, hypogeous species. the volume is dedicated to maria ławrynowicz botanist, mycologist, editor-in-chief of acta mycologica, on the occasion of her seventieth birthday and forty five years of scientific activity at the 56. congress of the polish botanical society in olsztyn, june 2013 photo m. petruszka in 1971 maria ławrynowicz received the degree of science doctor in biology (approximately equivalent to a phd) on the basis of her paper on fungal communi­ ties in oak­hornbeam forests of central poland. this event also marked the starting point of many years of cooperation with professor alina skirgiełło (university of warsaw) with the objective of developing an infrastructure for mycology. from this time, maria focused her attention on hypogeous fungi, one of her great early suc­ cesses being the discovery of black truffles (tuber mesentericum) in poland on the czestochowa upland, close to her birthplace. in 1984 maria obtained the scientific degree of habilitated doctor in biology (ap­ proximately equivalent to a dsc) on the basis of her treatise taxonomic-chorological study of european species of hypogeous ascomycetes, and in 1985 was appointed to the position of docent (reader) and then head of the mycological laboratory in the department. in 1994 maria obtained the scientific title of professor. in 2000, the mycological laboratory became the department of mycology within the chair of algology and mycology, which is now also led by maria. she is the author or a co­author of nearly 200 publications. her main research areas are: hypogeous fungi (establishing the northern limits of truffles in europe); spatial distributional zones of larger fungi in towns such as łódź; studies of fungal communities in nature reserves in central and other regions of poland; mycological monitoring in european oak forests (as coordinator for poland in a polish­czech­ italian project within a european union framework). she is also head of the her­ barium mycologicum, a fungal reference collection amounting now to over 30000 dried specimens and including one of the biggest contemporary collections of hypo­ geous fungi in europe. maria’s teaching activities have included: organization, programme development and leadership of the full­time doctoral studies of ecology and environmental protection, completed by 106 postgraduate students over a 20 year period; supervi­ sion of 9 doctorates; supervision of scientific aspects of over 100 masters theses. maria’s work in the conservation of nature, and particularly in fungal conserva­ tion deserves special attention. she was a co­author of the first list of protected fungi and the first red list of threatened fungi in poland. as far as duties in international organizations are concerned, she is the polish delegate to the european mycologi­ cal association, a member of the scientific council of the international society for conservation of fungi, and a member and, in the years 1995­1999 president of the european council for conservation of fungi. from 1990 to 1998 she was a member of the executive committee of the international mycological association. her con­ sequent active participation in meetings of these scientific bodies and, above all, in the thirteen congresses of european mycologists held since 1966, has meant that she is a well­known figure in the international mycological community. for many years professor ławrynowicz has also been a member of various important national scientific bodies including the committees of botany and nature conservation of the polish academy of sciences and, since 2004, the state council for nature conservation. maria ławrynowicz has been awarded several prizes and distinctions, including the bachelor’s cross of poland revival order, the medal of the national education committee, and honorary membership of the polish botanical society. krystyna czyżewska and andrzej grzywacz 2013-06-22t22:30:22+0100 polish botanical society book review błaszkowski j. 2012. glomeromycota. w. szafer institute of botany polish academy of sciences, kraków, p. 304. isbn: 978-83-89648-82-2 the monograph “glomeromycota” written by professor janusz błaszkowski is a unique publication in the mycological literature. this is the first worldwide comprehensive description of highly important soil fungi, which form arbuscular mycorrhizae (amf) with a vast majority of world’s plants (70– 90 %). although the fungal group is not numerous (ca. 220 species), its representative occur in any type of soil around the globe. the profound knowledge and detailed description provided in the monograph contribute to the author’s unquestionable success, which can be attributed to the impressive amount of work, determination, and perseverance as well as fascination with these fungi. a noteworthy fact is that mycorrhizal relationships had been discovered by a pole, franciszek kamieński (1881), over 130 years before the year of publication of the work (2012). the monograph comprises 137 species, which constitutes over half (62 %) of fungi from this group recognised so far. a great majority of the listed species originate from author’s own collections, and many of them are known exclusively from poland. the study material was collected from the rhizosphere of both crop plants and wild-growing vegetation. various techniques were used for identification of the fungi, including biochemical and molecular analyses, scanning electron microscopy as well as various techniques of light microscopy were employed for preparation of graphic documentation. the editorial aspect of the book reveals great diligence and accurateness, which is also visible in its large format (a4). the book provides the most relevant information about the history of the knowledge of these fungi, classification, research (field and laboratory) methods, and habitat and substrate preference. each species is fully described in terms of its morphological structures, basonyms, a full list of synonyms, and literature concerning its distribution range (in poland and worldwide). additionally, the phylogenetic position of the species has been discussed and lists of plants with which they form mycorrhizal relationships have been provided. another great value of the work is the excellent microphotographic documentation of microscopic acta mycologica vol. 48 (1): 133–134 2013 doi: 10.5586/am.2013.015 134 w mułenko preparations made by the author himself during verification or originating from other research units. the description sections are preceded by dichotomous keys for identification of fungi within successive taxonomic units – orders, families, and genera. these elements of the documentation make the work an ideal source of information for further diagnostics of fungi excluding the necessity of carrying out molecular analyses. the list of literature references is particularly extensive, as it includes both the oldest and the most recent papers. the final section presents very useful indexes of fungal, plant and locality names. generic or species taxa are additionally described by extremely interesting information concerning the etymology of these names. the work impresses with its considerable substantive value. the knowledge of species is a fundamental element in investigations of species diversity and recognition of mechanisms that rule phenomena occurring in the ecosystem. knowledge of fungi, particularly of soil fungal species is essential. in this group, the key role is played by mutualistic symbionts, including the glomeromycota described in the book. undoubtedly, the book will be helpful not only to mycologists, but also botanists and ecologists; therefore, it should find its place in every library of natural sciences. wiesław mułenko 2013-06-22t22:31:55+0100 polish botanical society cystogloea oelandica: an unusual new auricularioid species from sweden peter roberts mycology section, royal botanic gardens, kew, surrey tw9 3ab, uk p.roberts@rbgkew.org.uk r o b e r t s p.: cystogloea oelandica: an unusual new auricularioid species from sweden. acta mycol. 41 (1): 25 28, 2006. cystogloea oelandica, a new species and genus of auricularioid fungi (basidiomycota, incertae sedis), is described from the swedish island of öland. key words: basidiomycota, heterobasidiomycetes, parasitic fungi introduction whilst examining collections brought back from a field trip to the swedish island of öland in 2001, an undescribed auricularioid fungus was found with gelatinous basidiomes and unusual, deciduous probasidia arising from disarticulating hyphal compartments. this new species is described in a new genus, as follows: description of genus and species cystogloea p. roberts gen. nov. basidiomata gelatinosa. hyphae in matrice gelatinosa, moniliformes, disarticulantes, in typo generis efibulatae. probasidia ex hyphis disarticulatis constata, tenuitunicata. epibasidia cylindracea, septata. basidiosporae in typo generis oblongae. typus generis: cystogloea oelandica p. roberts the new genus is distinguished by producing gelatinous basidiomes in which auricularioid basidia arise from conspicuous probasidia which develop from disarticulated segments of moniliform hyphae. acta mycologica vol. 41 (1): 25-28 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 26 p. roberts cystogloea oelandica p. roberts sp. nov. basidiomata gelatinosa, hyalina, pustulata, 1 2 mm lata. hyphae in matrice gelatinosa, moniliformes, disarticulantes, efibulatae. probasidia ex hyphis disarticulatis constata, plerumque subglobosa vel ellipsoidea, c.15 20 x 12 17 µm, saepe oblonga vel cylindrica, fusiformes, c. 25 40 x 8 10 µm, tenuitunicata. epibasidia cylindracea, 45 60 x 5 7 µm, terseptata. sterigmata 4 10 (18) µm longa. basidiosporae oblongae, 8 10.5 x 5.5 6.5 µm. holotypus: sweden, öland, on quercus twig with pseudotrichia minor on old stromata of amphiporthe leiphaemia, 4 june 2001, b.m. spooner, k(m) 137630. basidiomes gelatinous, hyaline, erumpent, irregularly pustular, 1 – 2 mm across, often coalescing. hyphae in a soft, gelatinous matrix; disarticulating and only visible as disconnected, swollen, hyphal compartments in most basidiomes examined; where visfig. 1. cystogloea oelandica. (left to right) moniliform hyphae; disarticulated hyphal compart ments (probasidia) starting to produce epibasidia; mature, septate epibasidia arising from collapsed hyphal compartments (probasidia); basidiospores (type specimen). cystogloea oelandica: an unusual new auricularioid species from sweden 27 ible, composed of branching chains of short, swollen, hyaline compartments, 10 35 x 6 17 µm, 2.5 3.5 µm wide at septa, lacking clamp-connexions. hyphidia none seen. cystidia none seen. conidiophores none seen. basidia arising from disarticulated hyphal segments which act as probasidia; these probasidia thin-walled, variously shaped; mostly subglobose to ellipsoid, c.15 20 x 12 17 µm, but often fusiform, oblong to cylindrical, c. 25 40 x 8 10 µm, and sometimes irregular; the probasidia give rise to tubular, auricularioid epibasidia, 45 60 x 5 7 µm, which become triseptate at maturity. sterig mata 4 10 (18) µm long. basidiospores oblong (q = 1.4 1.6), 8 10.5 x 5.5 6.5 µm, germination not seen. fig. 1. the collection consists of several dozen, small, erumpent, gelatinous basidiomes which in the field were thought to be a species of tremella pers. the basidiomes were growing on an oak twig, most if not all arising from perithecia of pseudotrichia minor munk on old stromata of amphiporthe leiphaemia (fr.) butin. microscopically, all that can be seen in most mounts are loose probasidia in a gelatinous matrix, some of which are giving rise to tubular, auricularioid epibasidia bearing oblong basidiospores. examination of apparently immature basidiomes eventually revealed branched, rhizoctonia-like chains of swollen hyphal compartments. it appears that these swollen hyphal compartments readily disarticulate and subsequently act as probasidia. no known auricularioid species appears similar. deciduous basidia are a feature of the genus mycogloea l.s. olive, but in mycogloea species it is the tubular, epibasidial part of the basidium that is deciduous, the probasidia (produced on normal, narrow hyphae) remaining attached (b a n d o n i 1998). large globose to oblong probasidia are found in some species currently assigned to the genus cystobasidium (lagerh.) neuhoff, e.g. c. proliferans l.s. olive and the lichen associates c. hypogymniicola diederich & ahti and c. usneicola diederich & alstrup, but in these species the probasidia are produced on normal hyphae and are not deciduous (o l i v e 1952; d i e d e r i c h 1996). acknowledgment: many thanks to mariko parslow for kindly examining and identifying the ascomycetes associated with cystogloea oelandica. references b a n d o n i r.j. 1998. on some species of mycogloea. mycoscience 39: 31 36. d i e d e r i c h p. 1996. the lichenicolous heterobasidiomycetes. bibl. lichenol. 61: 1 98. o l i v e l.s. 1952. a new species of cystobasidium from new jersey. mycologia 64: 564 569. cystogloea oelandica: niezwykły, nowy, auricularioidalny gatunek ze szwecji s t r e s z c z e n i e praca zawiera opis rodzaju i gatunku cystogloea oelandica zebranego w 2001 roku na wy spach alandzkich w szwecji. 2014-01-01t11:43:15+0100 polish botanical society book reviews mirek z. (ed.). 2002-2009. biodiversity of poland. w. szafer institute of botany, polish academy of sciences, kraków „the book … assembling and presenting, for the first time in our history, the country’s species richness of plants, as well as fungi and other organisms traditionally included within the plant kingdom.” [from the foreword to the series by z. mirek]. the following volumes of the series biodiversity of poland were published in the years 2002-2009: vol. 1 flowering plants and pteridophytes of poland; vol. 2 ecological indicator values of vascular plants of poland; vol. 3 census catalogue of polish mosses; vol. 4 an annotated checklist of polish liverworts and hornworts; vol. 5 catalogue of polish prokaryotic and eukaryotic algae; vol. 6 the lichens, lichenicolous and allied fungi of poland. an annotated checklist; vol. 7 checklist of polish larger basidiomycetes; vol. 8 checklist of polish larger ascomycetes; vol. 9 a preliminary checklist of micromycetes in poland; vol. 10 myxomycetes of poland. a checklist; vol. 11 lichenicolous fungi of poland. a catalogue and key to species. acta mycologica vol. 45 (2): 247–250 2010 w. fałtynowicz. the lichens, lichenicolous and allied fungi of poland. an annotated checklist. vol. 6, pp. 435, 2003. isbn 83-89648-06-7. „the checklist is based on literature data and on herbarium materials partly. in the present list, 1768 accepted species from 357 genera are presented. among them, 1554 species from 273 genera are lichenized and 154 lichenicolous, and 60 non-lichenized species from 94 genera. besides, 160 infraspecific taxa (subspecies and varietas) are listed; 47 species which erroneously reported from poland are mentioned. for each taxon, both latin and polish names, synonyms, ecology, distribution in poland and literature reports concerning poland area provided. substrata from which polish specimens have been collected are described; for epiphytes, names of genera of all phorophytes were listed.” the main list is supplemented with index of accepted names of genera, index of synonyms and index of polish names. 248 ed. z. mirek w. wojewoda. checklist of polish larger basidio­ mycetes. vol. 7, pp. 812, 2003. isbn 83-89648-09-1. „about 400 genera and 2650 species known in poland are included in this book. some of them are invalid according to the icbn. some of the taxa are uncertainly classified, and other are mistakenly recorded from poland. the true number of basidiomycete taxa known from poland is about 2550. the genera and species are alphabetized. presently accepted latin names and the most important synonyms are given for each taxon. the affiliations to families and orders, and the number of species known in poland are given for the genera. the habitats in which the fungi grow, the plant community, substratum, mode of nutrition and fructification time are charactized. examples of species distribution in poland, including region form the most extensive part of the description of each species. polish and foreign taxonomic mycological literature together with illustrations of the fungi are cited.” index of polish names (genera and species of fungi) and appendix, close the checklist. m.a. chmiel. checklist of polish larger asco­ mycetes. vol. 8, pp. 152, 2006. isbn 83-89648-46-6. „this is the first comprehensive work on the larger ascomycetes found in poland. the checklist is based on data published in physiographical and mycological works. it contains 785 species listed alphabetically. for each species the accepted latin name is provided, followed by the name or names used in the source publications (synonyms) if they differ from the accepted names. the list also gives substrate descriptions and references to the source literature.” the main list is supplemented with index of authors’ names. biodiversity of poland 249 w. mułenko, t. majewski & m. ruszkiewiczmichalska (eds). a preliminary checklist of mi­ cromycetes in poland. vol. 9, pp. 752, 2008. isbn 978-83-89648-75-4. the taxa of the hyphochytriomycota, oomycota (kingdom chromista), chytridiomycota, glomeromycota, zygomycota, ascomycota, basidiomycota and the anamorphic fungi which belong mostly to the ascomycota (kingdom fungi) and plasmodiophoromycota (kingdom protozoa) found in poland are listed. additionally, the species of elaphomycetales, are also discussed. the checklist contains 5965 species. latin name, synonyms, substratum or the host species, references to the sources and occasionally notes, are given for each taxon. the orders in a framework of a particular phylum, and the species within orders are listed alphabetically. index of fungi close the checklist. a. drozdowicz, a. ronikier, w. stojanowska & e. panek. myxomycetes of poland. a checklist. vol. 10, pp. 103, 2003. isbn 83-89648-08-3. „this work is the first comprehensive checklist of the polish slime moulds (myxomycetes). it is based on published data and in principle only the source publications were taken into consideration. the list contains 222 species arranged alphabetically. for each species a valid latin name and a polish name are provided. below, the names used in source publications are listed, followed by the substrate description and references to source literature.” the main list is supplemented with index of latin names, index of polish names (with reference to latin names) and index of authors’ names. 250 ed. z. mirek k. czyżewska & m. kukwa. lichenicolous fungi of poland. a catalogue and key to species. vol. 11, pp. 133, 2009. isbn 978-83-89648-76-1. „the catalogue offers an overview of current knowledge of lichenicolous fungi in poland documented by publications on non-lichenized fungi, lichens and lichenicolous myxomycetes identified in poland in the period between 1851 and 2008. the list contains 249 species in 108 genera. lichenicolous non-lichenized fungi are represented by 216 species; 27 species are lichens starting their life cycle as parasites on lichens or found only facultatively as lichenicolous. six taxa of myxomycetes are reported as growing on lichens. fungi in the catalogue are arranged as follows. first is an alphabetical list of species, by genus and species within genera (synonyms, hosts, sources and notes, are given for species). all fungi belong to teleomorphic states of ascomycota (70 genera, 180 species), basidiomycota (5 genera, 12 species) and conidial fungi (28 genera, 51 species). the organisms occupied 271 lichen taxa (including 231 species). this is followed by the keys to lichenicolous fungi and slime moulds: general key, key 1 – lichenized fungi, key 2 – teleomorphic states of weakly lichenized or non-lichenized ascomycota, key 3 – basidiomycota, key 4 – conidial fungi or fungi with sclerotia only, and key 5 – mycetozoa found on lichens.” two indexes, index of hosts and their synonyms, with fungi found on their thalli, and index of synonyms, close the catalogue. compiled by mariusz hachułka 2014-01-01t11:51:43+0100 polish botanical society 268 a. bresinsky at the time when erast was born on october 23rd, 1928, in nõmme, a suburb of tallinn, as a son of a carpenter, estonia had been independ ent for just eight years. the young state then reorganized its educational system which had been under swedish, russian and german in fluence for a very long time. due to the forceful incorporation of estonia into the soviet union in 1940 and because of the final establishment of soviet power following the war, which had itself been imposed on the country by germany, the education of a young person had to be adapted according to the prevailing circumstances. erast was able to attend the gymnasium from which he graduated with exceptional success in 1947. it is surprising that he managed to complete his education in school without delay in a time of radical change and despite the turmoil caused by the war. he continued his education by study ing biology at the university of tartu until 1952. people outside estonia might not be aware of the fact that the swedish king gustav adolf had founded the university of tartu in 1632. the mycologist bondartsev (1877–1968) – the fungal genus bondarzewia was named after him – worked as a phytopathologist in leningrad (now renamed petersburg) at that time. he was also interested in the aphyllophorales of the soviet union. bondartsev had already collabo rated with the well known mycologist rolf singer and he gave erast parmasto the opportunity to continue his education in mycology using the modern methods available at that time. erast specialized in aphyllophorales and contributed substantially to the knowledge of this fungal group. in tartu he was awarded several aca demic ranks comparable to a doctoral degree (1955) and to the german “habilitation” (1969). in soviet times the estonians had to suffer from several constraints concerning their national in terests and their ethnic identity. on the positive side erast could travel through the vast areas of eurasia belonging to the soviet union – from the baltic satellite republics all the way to siberia and the far east (kamtschatka, sachalin), col lecting and researching fungi. he participated in a total of 63 mycological expeditions. in this way, over the decades, one of the largest collections of fungi in the whole of the soviet union was established in estonian tartu. this developed into a unique facility for research on the systematics of fungi and their distribution in the eurasian area, an area which is perhaps nowadays even more difficult to access than it was for a citizen of the soviet union in the times of the soviet union. professor erast parmasto during his lifetime erast remained connected with the same institute in estonia. he was af filiated with the institute of zoology and botany at the estonian academy of sciences from 1950 until his death. the institution changed its name several times over the years depending on the requirements of the times (nowadays its name is institute of agricultural and environmental sciences, estonian university of life sciences). the fungal herbarium (taam), where erast’s wife, ilmi parmasto is the curator, is located in this institute. erast’s professional employment was extensive, professor erast parmasto 269 ranging from gardener to scien tific assistant up to director of the institute of zoology and botany (1985–1990). since 1972 erast parmasto was a member of the estonian academy of sciences. between 1973 and 1981 he was also the academic secretary in the department of chemical, geological and biological sciences of the estonian academy of sciences. in addition to his affiliation with these institutions, erast was engaged as a professor of botany and ecology at the university of tartu (1987–1995) offering various lectures on mycology and scientific methodology. he served as the president of the estonian naturalist’s society (1973–1976) and was an honorary member of the society from 1988. it was a constant concern of erast to popularise mycological science. this was shown by his numerous popular articles on fungi and other topics, as well as his commitment as an edi tor (1958–1960) of the popular magazine eesti loodus (nature of estonia). the estonians al ways kept their national identity in spite of the constraints of foreign rule. this is demonstrated very well, not only by the national singing fes­ tivals but also by attention to art and ethnic traditions, identification with its own history and by the care of estonians for their countryside. in this connection it has to be mentioned that the first national park of the soviet union was established in estonia (lahemaa), despite the ambivalent position of the central government that did not recognize the word “national”. taking all of this into account it seems that the estonians were very well prepared to establish their independence once again when circum stances became favourable. erast parmasto played an important role in this process. on the occasion of the european mycological congress 1989 in tallinn, which he had organised, the participants from the west were welcomed in an unofficial manner by lennart meri, who was to become president of independent estonia. the presence of lennart meri to collection areas of fungi in the herbarium of tartu. blue dots = more than 5 expeditions, red dots = 2–4 expeditions, green = one expedition. erast parmasto participated in many of these expeditions and collected 37,000 samples out of 160,000 deposited in the herbarium of tartu. 270 a. bresinsky welcome some of the participants was achieved by the influence of his friend erast parmasto. it was the year of the human chain in the baltic states with the cry for freedom. erast parmasto’s high reputation both national and international came about as a result of several factors – professional competence and intelligence, the ability to work beyond ideologi cal boundaries and last but not least his remark able linguistic ability. his mother tongue was estonian, a finnish­ugric language. his popular scientific articles have mostly been published in this language. he also took care that myco logical publications in foreign languages were translated into estonian. in soviet times there was a considerable pressure to publish scientific papers predominantly in russian. in the course of liberalization and of the independence movement erast used english more and more as the language of science. erast was able to write as well as speak english fluently; therefore he had no difficulty in communicating with the “scien tific community” outside russia. he did this so well that he was appointed as an honorary member of the american society of mycology in 1993. his recognition in his own country could not have been better expressed than through his appointment as an honorary citizen of tartu and by his nomination as one of the 100 most important personalities in estonia. erast parmasto has left us the legacy of a large body of scientific work. a complete bibliogra phy has been published in folia cryptogamica estonica 49: 1–18, 2012. only few examples out of this list may be mentioned here. it was a special concern of erast to evaluate the size and variation of spore dimensions as important parameters for the delimitation of species within the hymenomycetes (parmasto, e., parmasto, i., 1987: variation of basidiospores in the hymenomycetes and its significance to their taxonomy. bibliotheca mycologica 115, 168 s.). this im portant study should be regularly consulted in taxonomic research on fungi, since spore size is almost always significant. as an example of his research on aphyllophorales of eurasia, his monograph on the lachnocladiaceae of the soviet union, published in 1970, has to be mentioned. he is also the author of an extensive study of the hymenochaetoid fungi of north america (mycotaxon 2001, 79: 107–176). out of the great number of publications on fungi in estonia, the checklist of fungi in this country and the distribution maps are noteworthy. he also established several data bases, for instance the one on corticioid fungi (cortbase). erast parmasto also contributed to the higher­level phylogenetic classification of fungi within an international team of scientists (hibbet, d.s. & al. 2007, mycol. res. 111: 509–547). the fungal genera erastia and parmastomyces have been named in honor of his efforts as a taxonomist. personally i got to know erast as a warm hearted, educated man, interested in many fields who made it possible for me to visit my country of birth after decades of absence. his achievements for mycology and for his country will last for a long time. andreas bresinsky (regensburg) [by courtesy of the editor in chief of folia cryptogamica estonica] for more information http://www.ut.ee/ial5/fce/fce49.html 2014-01-02t12:18:37+0100 polish botanical society entoloma jahnii, a rare species in europe joanna nita and mateusz stefaniak department of plant ecology and environmental protection, adam mickiewicz university umultowska 89, pl-61-614 poznań, malav@wp.pl, afanit@op.pl nita j., stefaniak m.: entoloma jahnii, a rare species in europe. acta mycol. 45 (2): 157–162, 2010. the authors describe two localities of entoloma jahnii wölfel & winterh., which were found during the mycocoenological research in pomerania and wielkopolska regions in the phytocoenoses of carici elongatae-alnetum and galio sylvatici-carpinetum associations. the article consists of a description of gathered specimens and some remarks concerning ecology of this rare species. key words: mycocoenology, pomerania region, wielkopolska region, carici elongataealnetum, galio sylvatici-carpinetum introduction entoloma jahnii is one of three members of section claudopus in poland (wojewoda 2003), which are characterized by pleurotoid or omphalinoid basidiocarps and excentric or absent stipe. it was described for the first time from germany by wölfel and winterhoff (1993). since then it was rarely found in several other european countries. in poland this species was found in two localities: lipka forest inspectorate (pomerania region) and krajkowo nature reserve (wielkopolska region). the aim of this paper it to contribute to the present knowledge on distribution and ecology of this species. acta mycologica vol. 45 (2): 157–162 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday entoloma jahnii 159 the collections were deposited at the herbarium of the department of plant ecology and environmental protection (pozm), the adam mickiewicz university, poznań. results the description refers to specimens from the first locality. characteristic features of the collected carpophores agree essentially with diagnoses given by noordeloos (2004). pileus pleurotoid (fig. 2), 1-8 mm wide, white, very thin (translucent), strongly pubescent, especially in the center, at the margin translucently striate and crenulated. stipe 1-3 mm long, 0.5-1 mm wide, excentric, sometimes rudimentary, bent, white and hairy. in the upper part slightly widened, at the base with long mycelium, which covers the substrate radially from the point of attachment. lamellae slightly or strongly decurrent, first white, then with pink tint, at maturity salmon pink, mixed, quite wide and thick. fig. 3. microscopic elements of e. jahnii: spores (a), capitate and cylindrical hyphae on the stipe (b) and the cap (c) surface. 160 j. nita and m. stefaniak basidia mostly 4-spored, rarely 2-spored, with clamps. cheiloand pleurocystidia absent. sporae 9.4-12.5 (15) (refers to the second locality) x 6.9-10 μm, variable in shape (fig. 3 a), isoand heterodiametric, 5 to 6 angled. the surface of stipe and cap with protruding capitate and cylindrical hyphae (fig. 3 b, c). clamp connections present. in lipka forest inspectorate during summer and autumn visits carpophores of entoloma jahnii were observed on a rotten log of betula (leg. j. nita). they were growing on the underside of the log, which was soaked in the boggy bottom, just after a periodical flood. the number of carpophores varied during the observation period of 3 years (8 visits) from 1 to over 100. during two observations carpophores of entoloma jahnii were accompanied by pluteus podospileus. in krajkowo reserve ten carpophores of this species were found on the small piece of wood which was part of an old and well decayed log of probably quercus robur or carpinus betulus lying on the ground among the dryopteris filix-mas and other plants. this was the moistest part of the plot. no other species of fungi were found on this piece of wood. the species was found only once, on july 29th 2009 (leg. m. stefaniak). discussion until recently, entoloma jahnii has been reported from poland only from the locality in lipka inspectorate forest. krajkowo nature reserve is the second locality of this species. this saprotrophic species has been noted in germany (krieglsteiner 2003), belgium (noordeloos 2004), denmark (læssøe 2008), finland (noordeloos 2008), sweden (ludwig 2007) and in the british isles (schafer 2008), where it was found on barked, rotten wood of frondose tree species: alnus, betula, populus, fraxinus and quercus. the species is regarded to be confined to moist and dry deciduous forests and copses (noordeloos 2004). it also shows the association with frondose tree species. this species was found by the authors in two forest associations which offer different ecological conditions, the main differences are moisture conditions and availability of wood of different tree species. the locality in lipka forest inspectorate was temporarily flooded, whereas krajkowo nature reserve is rather dry and never flooded. the first locality offered also different substrates (alnus glutinosa, betula sp.) compared to krajkowo nature reserve (quercus robur, carpinus betulus, ulmus leavis). the localities show full scale of substrates and ecological conditions in which this species can occur – from moist to mesophilous forest associations. entoloma jahnii is red-listed in the mecklenburg-vorpomernn region (schwik et al. 1999), and in denmark (the danish red data book 2010). fruitbodies of this species are frequently found in hidden places, such as the underside of logs, branches, or pieces of wood (noordeloos 2004), and can be easily overlooked. this feature seems to be one of the reasons of its rarity, nevertheless in our opinion this species is really rare in poland. in recent years many studies on macrofungi were conducted in broadleaved forests and copses (e.g., bujakiewicz 1989; bujakiewicz et al. 2005; bujakiewicz, stefaniak 2009; friedrich 1994; kujawa 2009; lisiewska, malinger 2001; ławrynowicz entoloma jahnii 161 et al. 2002; nita, bujakiewicz 2005) in which, according to literature and the author’s own observations suitable ecological conditions occur; still the species is known only from two localities. in our opinion it would be appropriate to include this species on the polish red list (wojewoda, ławrynowicz 2006) with the category e. acknowledgements. authors would like to thank prof. anna bujakiewicz for her valuable comments and discussion on our manuscript. we are grateful mr. mieczysław nita for his kind permission to use his photographs and miss magdalena stefaniak for improving the language. we are also indebted to the anonymous reviewer for useful suggestions that helped to improve our work. the study was supported by ministry of science and higher education, grants nos. n304 060 31 and n304 0436 39. references bujakiewicz a. 1989. macrofungi in the alder and alluvial forests in various parts of europe and north america. opera bot. 100: 29–41. bujakiewicz a., nita j., bałazy s. 2005. first localities of the recently described fungus – cordyceps bifusispora o. e. ericksson. acta mycol. 40 (2): 251–258. bujakiewicz a., stefaniak m. 2009. udział macromycetes w fitocenozach leśnych rezerwatu ,,las liściasty w promnie” (nadleśnictwo czerniejewo). bad. fizjogr. pol. zach., b, 58: 137–170. friedrich s. 1994. charakterystyka socjologiczno-ekologiczna mikoflory zbiorowisk roślinnych cedyńskiego parku krajobrazowego. akademia rolnicza w szczecinie, rozprawy 161: 1–100. kujawa a. 2009. macrofungi of wooded patches in the agricultural landscape. i. species diversity. acta mycol. 44 (1): 49–75. krieglsteiner g. j. (ed.) 2003. die großpilze baden-württembergs. band 4 ständerpilze: blätterpilze ii. verlag eugen ulmer, stuttgart. ławrynowicz m., dziedziński t., szkodzik j. 2002. macrofungi of aceri-tilietum and tilio-carpinetum in the „dolina rzeki brdy“ nature reserve in the bory tucholskie (nw poland). acta mycol. 37 (1/2): 63–76. lisiewska m., malinger m. 2001. macromycetes w różnych postaciach grądu środkowoeuropejskiego na terenie uroczyska marcelin w poznaniu. bad. fizjogr. pol. zach. b, 50: 7–40. ludwig e. 2007. pilzkompendium. band 2. beschreibungen. fungicon verlag, berlin. læssøe t. 2008. muslinge-rødblad (entoloma jahnii) nu også i danmark svampe 57: 34–35. nita j., bujakiewicz a. 2005. grzyby wielkoowocnikowe w fitocenozach łęgu wiązowego querco-ulmetum minoris i olsu carici elongatae-alnetum w lesie złotowskim (pomorze zachodnie). bad. fizjogr. pol. zach. ser. b, 54: 7–33. nita j., bujakiewicz a. 2007. łęgi i olsy ostoją rzadkich i zagrożonych grzybów wielkoowocnikowych. studia i materiały centrum edukacji przyrodniczo-leśnej. siedliska i gatunki wskaźnikowe w lasach. 9 (2/3): 519–529. nita j., bujakiewicz a. 2009. riparian and alder forests – shelters for rare and threatened macromycetes (in:) z. mirek, a. nikel (eds). rare, relict and endangered plant and fungi in poland. w. szafer institute of botany, polish academy of sciences, kraków: 325–334. noordeloos m. e. 2004. fungi europaei 5a. entoloma s.l. supplemento. edizioni candusso. italia. noordeloos, m. e. 2008. entoloma (fr.) p.kumm. (in:) h. knudsen, j. vesterholt (eds). funga nordica. agaricoid, boletoid and cyphelloid genera. nordsvamp-copenhagen. schwik j., westphal b., bütow r., michael h., richter k., schurig b. 1999. rote liste der gefährdeten großpilze mecklenburg-vorpommerns. 2. fassung. stand: november 1999. umweltministerium des landes mecklenburg-vorpommern, schwerin. schafer d. 2008: persistence pays off entoloma jahnii, a new british record. field mycology: 9 (2): 49–50. the danish red data book. national environmental research institute, http://www2.dmu.dk/ wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland 7: 1–812. w. szafer institute of botany, polish academy of sciences, kraków. wojewoda w., ławrynowicz m. 2006. red list of the macrofungi in poland. (in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. 3. ed.: 53–70. w. szafer institute of botany, polish academy of sciences, kraków. wölfel g., winterhoff w. 1993. entoloma jahnii, ein neuer holzbewohner. österr. z. pilzk. 2: 11–14. 162 j. nita and m. stefaniak entoloma jahnii, gatunek rzadki w europie streszczenie artykuł zawiera informację na temat dwóch stanowisk entoloma jahnii wölfel & winterh. w polsce. gatunek ten notowano w nadleśnictwie lipka (pojezierze krajeńskie) oraz w rezerwacie przyrody „krajkowo” (wielkopolska), w fitocenozach carici elongatae-alnetum oraz galio sylvatici-carpinetum, podczas wieloletnich badań mikocenologicznych na stałych powierzchniach badawczych. w pracy zawarto szczegółowy opis i ilustracje cech zebranych owocników tego gatunku, oraz uwagi dotyczące jego rozmieszczenia i ekologii. autorzy podkreślają powiązanie e.jahnii z lasami i zaroślami liściastymi, oraz proponują ujęcie tego gatunku na czerwonej liście grzybów wielkoowocnikowych w polsce. 2014-01-01t11:51:07+0100 polish botanical society mycorrhizal inoculation of pecan seedlings with some marketable truffles gian maria niccolò benucci¹, gregory bonito², leonardo baciarelli falini¹, mattia bencivenga¹ and domizia donnini¹ 1department of applied biology, university of perugia, borgo xx giugno 74 i-06121 perugia, gian.benucci@gmail.com ²biology department, duke university, durham, nc 27708, usa benucci g.m.n., bonito g., baciarelli falini l., bencivenga m., donnini d.: mycorrhizal inoculation of pecan seedlings with some marketable truffles. acta mycol. 47 (2): 179–184, 2012. pecan is the common name of carya illinoinensis (wangenh.) k. koch, an ectomycorrhizal tree native to north america, also frequently known as hickory. mycorrhizal inoculations of pecan seedlings with: tuber aestivum vittad., t. borchii vittad., t. indicum cooke & massee, and t. lyonii butters are described and discussed. key words: carya illinoinensis, truffle spore-slurry, multi-cropping, tuber borchii, t. aestivum, t. indicum, t. lyonii introduction pecan is the common name of carya illinoinensis (wangenh.) k. koch, also frequently known as hickory. it is a deciduous tree in the juglandaceae family with pinnately compound leaves, which produces large and economically valuable nuts. this tree also establishes ectomycorrhizal (ecm) associations with hypogeous and epigeous fungi. pecan is native to north america and grows naturally on the moist bottomland habitat along streams in the usa ranging from indiana south to kentucky and alabama and from iowa south to texas, principally along the mississippi river. this species is cultivated largely in southeastern north america exclusively for the edible nuts, which are used for pecan pie, pecan pralines or food products including cereals, energy bars and candy bars. pecan production of the united states it is likely to be over 433 million dollar per year, and the industry is in a period of growth (usda 2008). a large proportion of pecan harvests are being exported to china, a rapidly growing market. large productive commercial pecan orchards have also been recently established in brazil, israel, and australia (wakeling et al. 2001). acta mycologica vol. 47 (2): 179–184 2012 180 g.m.n. benucci et al. in the late 1980’s hanlin and colleagues (1989) discovered truffles fruiting naturally in pecan orchards in georgia, usa. bonito and colleagues (2011) followed up on this study and showed that the ecm truffle genus tuber (pezizaceae) is naturally abundant in fungal communities of many pecan orchards in georgia, including the species tuber lyonii butters, whose ascocarps are frequently found and collected. although t. lyonii is sold and served in us restaurant with success, the market is still underdeveloped because of its low organoleptic qualities, low availability and low quality of truffles collected (bonito et al. 2011; benucci et al. 2012a). truffles are harvested by raking, then mature and less aromatic immature truffles are marketed spreading a negative opinion on the product. material and methods to overcome these problems and evaluate whether pecan can associate with both native and non-native truffle species as first step of future potential multi-cropping of pecans and truffles, some investigation on ecm inoculations with tuber spp. on pecan have been realized (bonito et al. 2010; bonito et al. 2011; benucci et al. 2012a; bonito et al. 2012). spore inocula were prepared with appreciated and widely distributed truffle species: tuber aestivum vittad., tuber borchii vittad., tuber indicum cooke & massee, tuber macrosporum vittad. and tuber lyonii butters. after inoculations with spore slurries, pecan seedlings were grown in greenhouse conditions for several months (fig. 1a), after which plants were harvested and mycorrhization levels quantified visually (bencivenga et al. 1995). ecm were described morpho-anatomically and molecularly by species-specific polymerase chain reactions (pcrs) (mello et al. 1999; mello et al. 2002) and/or by sequencing and analyzing the internal transcribed spacer (its) (white 1990) and 28s large subunit (lsu) (vilgalys, hester 1990) nuclear ribosomal dna (nrdna) region. results tuber borchii and t. aestivum formed abundant ecm on pecan (fig. 1c, 1e), with ≈60% and ≈40% root-tip colonization, respectively (benucci et al. 2012a). in contrast, no ecm of t. macrosporum were detected on pecan, which may indicate that this host is not compatible with this fungal species, although it is known that t. macrosporum ecm are difficult to obtain under laboratory and nursery conditions (benucci et al. 2012b). morphological and anatomical characteristics of t. borchii and t. aestivum on pecan have similar features to those that they display with other host species (e.g., oak, hazelnut), and their morphologies are consistent with previous reports (giomaro et al. 2000; granetti et al. 2005). in particular, t. borchii possessed straight needle-shaped cystidia and puzzle-like cell pattern (fig. 1g, 1h), while t. aestivum possessed curly cystidia and angular cell pattern (fig. 1f, 1i). mycorrhizal inoculation 181 fig. 1. pecan seedlings and morphology of t. aestivum, t. borchii, t. indicum and t. lyonii ecm. (a) pecan seedlings in the nursery; (b) t. lyonii ecm on pecan roots (scale bar=700 μm); (c) t. borchii ecm on pecan roots (scale bar=700 μm); (d) t. indicum ecm on pecan roots (scale bar=700 μm); (e) t. aestivum ecm on pecan roots (scale bar=700 μm); (f) curly cystidia of t. aestivum ecm (scale bar=30 μm); (g) outer mantle layer of t. borchii ecm (scale bar=30 μm); (h) needle-shape cystidia of t. borchii (scale bar=30 μm); (i) outer mantle layer of t. aestivum ecm (scale bar=30 μm); (j) outer mantle layer of t. lyonii ecm (scale bar=30 μm). 182 g.m.n. benucci et al. the molecular pcr assay confirmed that amplicons obtained from truffle ecm are consistent with our morphological and taxonomical assessment. their its nrdna sequences matched at 100% query coverage with 0.0 e-values and 99-100% identity to those present in genbank for the same fungal species. bonito and colleagues (2010) were able to synthesize t. indicum ecm on carya illinoinensis seedlings using standard inoculation methods. the identification of ecm was confirmed by morphology (fig. 1d) and its sequence data. ecm produced by t. indicum on pecan were unramified to irregularly pinnate, dark amber in colour and showed characteristic puzzle-like pseudoparenchyma cells in the outer mantle with emanating right-angle branching cystidia, as reported from other hosts (geng et al. 2009). tuber lyonii ecm were also well formed on pecan seedlings 6 months after they had been inoculated with t. lyonii spores (bonito et al. 2011). ectomycorrhizas are characterized by a thin and smooth mantle with epidermoid puzzle-like cells (fig. 1b, 1j). molecular its sequences confirmed the identity of t. lyonii ecm. discussion together these data demonstrate that european, asian and american truffles are able to form healthy ecm on pecan, regardless of the fact that this plant is endemic to north america. these studies lead us to conclude that there is the potential to multi-crop various truffle species with pecan. to successfully cultivate pecan with the simultaneous production of truffles represents a unique opportunity to diversify agricultural outputs from a given tract of land, and to add extra income in an environmental and sustainable way (benucci et al. 2013; donnini et al. 2013). furthermore, given the attributes of pecan wood for lumber and flooring, opportunities exist to obtain additional revenue streams from the timber production at the end of the cultivation cycle or following stand thinning (benucci et al. 2012a). in us, although some orchards were originally brought for lumber and furniture, most pecan trees taken out are chipped and used for smoking meats in fires and traditional barbeques. however, the wood is quite dense, making it hard on saw blades of modern mill equipment, and transport costs are high. nonetheless, once co2 is regulated in the us, we expect more sustainable practices will develop and pecan wood and timber will be favored again for flooring and furniture. conclusions in conclusion, given the ability of pecan to readily for ecm with economic truffle species from five different clades tuber that includes north american, european, and asian truffle species, pecan holds promise as a host plant that is compatible with the co-cropping of truffles and nuts. whether or not tuber ecm are maintained mycorrhizal inoculation 183 when mycorrhized seedlings are planted out in an orchard setting, and whether these truffle species will fruit with pecan still need to be addressed however. moreover, in humid climates pecan cultivation often requires heavy inputs of biocides to combat plant diseases and inputs of fertilizers to boost the production of nuts. further studies are needed to address the potential accumulation of these harmful substances inside of truffles. acknowledgements. suggestions and critical comments on the manuscript made by anonymous reviewers are greatly acknowledged. the authors are grateful to andrea vece and andrea gógán csorbainé, who provided part of the truffles needed for mycorrhization trials. references bencivenga m., donnini d., tanfulli m., guiducci m. 1995. tecnica di campionamento delle radici e degli apici radicali per la valutazione delle piante micorrizate. micol ital 2: 35-47. benucci g. m. n., bonito g., baciarelli falini l., bencivenga m. 2012a. mycorrhization of pecan trees (carya illinoinensis) with commercial truffle species: tuber aestivum vittad. and tuber borchii vittad. mycorrhiza 22: 383-392. doi 10.1007/s00572-011-0413-z benucci g. m. n., bonito g., baciarelli falini l., bencivenga m., donnini d. 2013. truffles, timber, food, and fuel: sustainable approaches for multi-cropping truffles and economically important plants. edible ectomycorrhizal mushrooms,(zambonelli a, bonito g, eds), springer-verlag berlin heidelberg. benucci g. m. n., gogan csorbai a., baciarelli falini l., bencivenga m., di massimo g., donnini d. 2012b. mycorrhization of quercus robur l., quercus cerris l. and corylus avellana l. seedlings with tuber macrosporum vittad. mycorrhiza 22: 639-646. doi 10.1007/s00572-012-0441-3 bonito g., brenneman t., vilgalys r. 2011. ectomycorrhizal fungal diversity in orchards of cultivated pecan (carya illinoinensis; juglandaceae). mycorrhiza 21: 601-612. doi 10.1007/s00572-011-0368-0 bonito g., trappe j. m., donovan s., vilgalys r. 2010. the asian black truffle tuber indicum can form ectomycorrhizas with north american host plants and complete its life cycle in non-native soils. fungal ecology 4: 83-93. doi 10.1016/j.funeco.2010.08.003 bonito g., smith m. e., brenneman t., vilgalys r. 2012. assessing ectomycorrhizal fungal spore banks of truffle producing soils with pecan seedling trap-plants. plant and soil 356: 357-366. doi 10.1007/ s11104-012-1127-5 donnini d., gargano m. l., perini c., et al. 2013. wild versus cultivated mushrooms as a model of sustainable development. plant biosystems. doi 10.1080/11263504.2012.754386 geng l. y., wang x. h., yu f. q., et al. 2009. mycorrhizal synthesis of tuber indicum with two indigenous hosts, castanea mollissima and pinus armandii. mycorrhiza 19: 461-467. giomaro g., zambonelli a., sisti d., cecchini m., evangelista v., stocchi v. 2000. anatomical and morphological characterization of mycorrhizas of five strains of tuber borchii vittad. mycorrhiza 10: 107-114. granetti b., de angelis a., materozzi g. 2005. umbria terra di tartufi. umbriagraf, terni, italy. hanlin r. t., wu m. l., brenneman t. b. 1989. the occurrence of tuber texense in georgia. mycotaxon 34: 387-394. mello a., garnero l., bonfante p. 1999. specific pcr-primers as a reliable tool for the detection of white truffles in mycorrhizal roots. new phytologist 141: 511-516. mello a., cantisani a., vizzini a., bonfante p. 2002. genetic variability of tuber uncinatum and its relatedness to other black truffles. environ microbiol 4: 584-594. usda 2008. national agricultural statistics service. noncitrus fruits and nuts: fr nt 1–3 (09). unites states, http://usda.mannlib.cornell.edu/mannusda/viewdocumentinfo.do?documentid=1 vilgalys r., hester m. 1990. rapid genetic identification and mapping of enzymatically amplified ribosomal dna from several cryptococcus species. journal of bacteriology 172: 4238-4246. 184 g.m.n. benucci et al. wakeling l. t., mason r. l., bruce r., caffin n. a. 2001. composition of pecan cultivars wichita and western schley [carya illinoinensis (wangenh.) k. koch] grown in australia. journal of agricultural and food chemistry 49: 1277-1281. doi: 10.1021/jf000797d white t. j. 1990. amplification and direct sequencing of fungal ribosomal rna genes for phylogenetics. pcr protocols, a guide to methods and applications,(innis ma, gelfand dh, sninsky jj, white tj, eds), academic press inc., new york , usa pp. 315-322. 2014-01-02t12:13:52+0100 polish botanical society rinodina griseosoralifera, a lichen species new to the western carpathians paweł czarnota1 and martin kukwa2 1scientific laboratory of the gorce national park, poręba wielka 590 pl-34-735 niedźwiedź, pawel.czarnota@gpn.pl 2department of plant taxonomy and nature protection, university of gdańsk al. legionów 9, pl-80-441 gdańsk, dokmak@univ.gda.pl c z a r n o t a p., k u k w a m.: rinodina griseosoralifera, a lichen species new to the western carpathians. acta mycol. 42 (2): 287-290, 2007. rinodina griseosoralifera coppins is reported for the first time from poland and the western carpathians. it is known there from the gorce mts, only locality up to now. details of the chemistry, morphology and general distribution are provided and similar taxa are discussed. key words: rinodina, sorediate lichens, atranorin, zeorin, carpathians, poland introduction lichen genus rinodina (ach.) gray is comprised of crustose lichens with leca-(ach.) gray is comprised of crustose lichens with leca-comprised of crustose lichens with lecanorine apothecia and brown, characteristically septate spores (for full description see n o w a k 1998; m a y r h o f e r , m o b e r g 2002). most of the taxa reproduce by spores, but a few possess vegetative soredia and isidia; these are usually sterile and usually difficult to identify since many other, even unrelated, species are superficially similar. in such cases, secondary chemistry is invaluable in their determination (e.g. tø n s b e r g 1992). in poland, only two sorediate epiphytic rinodina species, r. colobina (ach.) th. fr. and r. efflorescens malme, have been reported to date (fa ł t y n o w i c z 2003). during the identification of some sterile crustose lichens we came across a specimen resembling r. efflorescens, but thin layer chromatography revealed the presence of atranorin and zeorin. the chemistry and morphology matches r. griseosoralifera coppins, a taxon not known from our country. this paper presents the first record of r. griseosoralifera for poland and the western carpathians, provides information on its chemistry, morphology and general distribution, and discusses its affinities to several sorediate, usually sterile, taxa. acta mycologica vol. 42 (2): 287-290 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 288 p. czarnota and m. kukwa material and methods morphology of the specimen was studied with the aid of a stereomicroscope and chemical analyses were carried out using thin layer chromatography (tlc) according to o r a n g e et al. (2001). the material is stored in gpn, with a duplicate donated to ugda-l. results rinodina griseosoralifera coppins – lichenologist 21: 169. 1989 morphology and chemistry. r. griseosoralifera is a crustose lichen forming an episubstratal thallus and consisting of greenish-white to brown areoles which usually entirely dissolve into soredia, as in the polish collection. soralia are blue-grey, formed at the top of areoles, discrete (fig. 1a) or forming a more or less continuous sorediate crust (fig. 1b). apothecia were not observed in the polish collection. the species produces atranorin and zeorin. tø n s b e r g (1992) also reported possible traces of unidentified terpenoids, but we did not detect any additional substances. affinities. due to its blue-grey external soredia r. griseosoralifera is superficially similar to several taxa: buellia griseovirens (turner & borrer ex sm.) almb., caloplaca chlorina (flot.) sandst., rinodina colobina and r. efflorescens. the most significant character is the chemistry. b. griseovirens produces norstictic acid and atranorin as major compounds (squash preparation reacts k+ yellow turning orange and forming needle-like crystals), c. chlorina does not contain lichen substances, r. colobina has substances called ‘colobina unknows’ in the thallus, and r. efflorescens has pannarin (with additional substances) causing p+ orange-red reaction of soralia (tø n s b e r g 1992). additional characters differentiating these taxa are shown in the table 1. ecology. in the polish locality the species covered eutrophic bark and occasionally mosses at the base of an old pyrus, forming large patches up to ca 10 dm2. it was associated with acrocordia gemmata (ach.) a. massal., bacidia subincompta (hoffm.) a. massal., caloplaca obcurella (j. lahm) th. fr., candelariella xanthostigma (ach.) lettau, lepraria vouauxii (hue) r. c. harris, normandina pulchella (borrer) nyl. and phaeophyscia endophoenicea (harm.) moberg. the polish habitat corresponds to that described by pa l i c e (1999), suggesting that it may by a more frequent species in old orchards or solitary trees in poland, but overlooked due to its usually sterile form and the necessity to examine it by tlc. distribution. in poland, r. griseosoralifera is known from a single locality in the gorce mts, but it may be more widely distributed since suitable habitats are common in the country. in europe, the species is rather uncommon, but widely distributed. to date, it has been reported from austria and britain (c o p p i n s 1989), ireland (s e a w a r d 1994), czech republic (pa l i c e 1999), germany (s c h o l z 2000), the netherlands (a p t r o o t et al. 2001), norway (c o p p i n s 1989; tø n s b e r g 1992), spain (l l i m o n a , h l a d u n 2001), switzerland (d i e t r i c h , s c h e i d e g g e r 1996) and ukraine (eastern rinodina griseosoralifera 289 carpathians) (k o n d r a t y u k , c o p p i n s 2000). outside europe, it is known from the canary islands (tø n s b e r g 2002) and north america (tø n s b e r g 1993). polish specimen examined. western beskidy mts, gorce mts, poręba wielka village, old wodzicki’s manor park, 49°37’02”n/20°03’57”e, alt. 530 m, on bark of old pyrus communis in old fruit orchard, 18.04.2007, leg. p. czarnota 5147 (gpn, duplicate in ugda-l–14058). additional specimen examined. norway, hordaland, bergen, arna, arna church, utm 32v ln 0503, alt. 10 m, on acer platanoides, 22.07.1990, leg. t. tønsberg 13338 (bg l-24513). ta b l e 1 comparison of rinodina griseosoralifera and superficially similar taxa occurring in poland (acc. to tø n s b e r g 1992 and author’s studies) feature taxa rinodina griseosoralifera rinodina efflorescens rinodina colobina caloplaca chlorina buellia griseovirens non-soraliate parts of thallus indistinct, small areolae, early dissolving into soralia or sorediate crust areolate to subsquamulose, often dissolving into soralia areolate to warted, ±disappearing, early becoming sorediate mostly quite thick, at least partially distinctly areolate to ±leprose mostly distinct, areolate to more or less continuous prothallus indistinct, brown visible around the areolae indistinct, between areolae, sometimes brownish black absent sometimes distinct, brownish to brownish violet usually present, brownish, often bluish tinged colour of soralia bluish-grey to pale greyish green brown, greyish brown, pale to dull greenish yellow grey-black to bluish-grey, rarely greyish green grey to bluish grey greyish white, yellowish, greenish to dark grey, often with a bluish tinge shape of soralia ±circular, plane to convex, discrete or confluent, bursting from areolae punctiform, ±flat to hemispherical bursting from areolae or substratum not well defined, mostly irregular and efflorescent, sometimes marginal at areolae marginal at areolae, punctiform to confluent forming leprose sorediate crust mostly circular, convex, plane or crateriform chemistry (major compounds) atranorin, zeorin; k–, p– or ±yellowish, c– pannarin, efflorescens unknown (pigment), ±zeorin; k–, p+ red, c– ‘colobina unknowns’; at least partially k± violet, p–, c– no substances; at least partially k± violet, n± violet, p–, c– atranorin, norstictic acid ±griseovirens unknowns; k+ yellow (forming crystals), p+ yellow, c– or ±yellowish pigmentation of external soredia bluish-grey, k+ brown, n– often brown, k+ fuscous brown, n– grey-black, k+, c+, n+ violet bluish, k+ violet, n+ violet brown, often bluish tinged, k+ fuscous brown, n– 290 p. czarnota and m. kukwa acknowledgements. we are indebted to an anonymous reviewer for suggestions on the manuscript, as well as to professor mark seaward (bradford, uk) for english correction and some improvements. references a p t r o o t a . , s p a r r i u s l . , v a n h e r k k . , d e b r u y n u . 2001. origin and distribution of recently described lichens from the netherlands. aktuelle lichenologische mitteilungen 5: 13–25. c o p p i n s b . j . 1989. rinodina griseosoralifera, a new corticolous sorediate lichen from western europe. lichenologist 21: 169–172. d i e t r i c h m . , s c h e i d e g g e r c . 1996. the importance of sorediate crustose lichens in the epiphytic lichen flora of the swiss plateau and the pre-alps. lichenologist 28: 245–256. fa ł t y n o w i c z w. 2003. the lichens, lichenicolous and allied fungi of poland – an annotated checklist. w. szafer institute of botany, polish academy of sciences, kraków, 435 pp. k o n d r a t y u k s . ya . , c o p p i n s b . j . 1999. basement for the lichen monitoring in uzhansky national nature park, ukrainian part of the biosphere reserve ‘eastern carpathians’. roczniki bieszczadzkie 8: 149–192. l l i m o n a x . , h l a d u n n . l . 2001. checklist of the lichens and lichenicolous fungi of the iberian peninsula and balearic islands. bocconea 14: 1–581. m a y r h o f e r h . , m o b e r g r . 2002. rinodina. nordic lichen flora 2: 41–69. n o w a k j . 1998. porosty (lichenes). vi. 2. buelliaceae. (in:) rośliny zarodnikowe polski i ziem ościennych. instytut botaniki im. w. szafera pan, kraków, 236 pp. o r a n g e a . , j a m e s p. w. , w h i t e f. j . 2001. microchemical methods for the identification of lichens. british lichen society, london, 101 pp. pa l i c e z . 1999. new and noteworthy records of lichens in the czech republic. preslia 71: 289–336. s c h o l z p. 2000. katalog der flechten und flechtenbewohnenden pilze deutschlands. schriftenreihe für vegetationskunde 31: 1–298. s e a w a r d m . r . d . 1994. vice-county distribution of irish lichens. proceedings of the royal irish academy 94 b: 177–194. tø n s b e r g t. 1992. the sorediate and isidiate, corticolous, crustose lichens in norway. sommerfeltia 14: 1–331. tø n s b e r g t. 1993. additions to the lichen flora of north america. bryologist 96: 138–141. tø n s b e r g t. 2002. halecania viridescens and rinodina griseosoralifera new to africa from the canary islands. graphis scripta 13 (2): 52–54. rinodina griseosoralifera, gatunek porostu nowy dla karpat zachodnich s t r e s z c z e n i e rinodina griseosoralifera jest nadrzewnym porostem skorupiastym wytwarzającym soredia. plecha tego gatunku składa się z areolek, z których większość przekształca się w oddzielone od siebie (fig. 1a) lub zlewające się (fig. 1b) soralia; soredia mają kolor niebiesko-szary. obecność wtórnych metabolitów, atranoryny i zeoryny, pozwala odróżnić ten gatunek od innych, morfologicznie bardzo podobnych ‘sorediowanych’ przedstawicieli rodzaju rinodina, tj. r. colobina i r. efflorescens, jak również od buellia griseovirens i nadrzewnych form caloplaca chlorina. cechy chemiczne i morfologiczne ułatwiające identyfikację sterylnych form tych pięciu gatunków przedstawiono w tabeli 1. r. griseosoralifera jest nowym składnikiem bioty porostów polski i jednocześnie karpat zachodnich. pierwsze jego stanowisko zostało znalezione w gorcach, w starym sadzie podworskim we wsi poręba wielka. gatunek ten rośnie na korze pyrus communis, a towarzyszą mu m.in. acrocordia gemmata, bacidia subincompta, caloplaca obscurella, normandina pulchella i phaeophyscia endophoenicea. fig. 1. habit of rinodina griseosoralifera (czarnota 5147, gpn); a – distinct soralia; b – confluent soralia; scale bars = 1 mm. 2014-01-01t11:46:49+0100 polish botanical society new records of erysiphales and uredinales from poland agata wołczańska department of botany and mycology, institute of biology, maria curie-skłodowska university akademicka 19, pl-20-033 lublin, agata.wolczanska@poczta.umcs.lublin.pl wołczańska a.: new records of erysiphales and uredinales from poland. acta mycol. 43 (1): 71–75, 2008. puccinia mei-mamillata semad. was found for the first time in poland on angelica sylvestris l., neoerysiphe galeopsidis (dc.) u. braun on melittis melisophyllum l. and podosphaera xanthii (castagne) u. braun & n. shishkoff on physalis alkekengi l. the new collections are described, illustrated and discussed herein. the key to the aecial state of rust fungi on angelica in poland is provided. another four species are known from few localities on their hosts in poland. key words: rust fungi, powdery mildews, distribution introduction in the last few years there have been an increase in the number of reports on the occurrence of new species of fungi in poland as well as on the extending range of native species and their invasion onto new hosts. this especially relates to powdery mildew (erysiphales), but there are also data on uredinales, ustilaginomycetes and anamorphic fungi (e.g. adamska 2001, 2005; piątek 2003a,b, 2005; piątek, wołczańska 2004; ruszkiewicz-michalska 2006; ruszkiewicz-michalska, michalski 2005; wołczańska 2007; wołczańska, oklejewicz 2001; wołczańska, lamorski 2006; wołczańska, rozwałka 2005). this phenomenon may stem from natural migration of fungi or the introduction thereof into new areas, from migration of hosts and climatic changes, favorably enabling phytopathogens to develop fully. the present paper adds other plants to the list of hosts of phytopathogenic fungi in poland and provides new data to their distribution. detailed descriptions and illustrations are given for the species observed on new hosts: puccinia mei-mamillata semad. on angelica sylvestris, neoerysiphe galeopsidis (dc.) u. braun on melittis melisophyllum and podosphaera xanthii (castagne) u. braun & n. shishkoff on physalis alkekengi. all the collected specimens have been deposited in the mycological herbarium of the department of botany and mycology, university of maria curie-skłodowska in lublin (lbl m). the taxonomy and nomenclature of uredinales are based on the acta mycologica vol. 43 (1): 71–75 2008 72 a. wołczańska monograph by majewski (1977, 1979), and powdery mildews on the monograph and papers by braun (1987, 1995, 1999) and braun & takamatsu (2000). the names of plants are taken after mirek et al. (2002). list of species erysiphales neoerysiphe galeopsidis (dc.) u. braun [= erysiphe galeopsidis dc.] white, dense, persistent mycelium on the leaves. appresoria lobed. conidia ellipsoid-ovoid: 30-38(40) x 12-18 μm, produced in chains (fig. 1a, b). specimen examined: on melittis melisophyllum: roztocze national park, maziarki reserve, 25.06.1998, lbl m-8650. remarks. this species was collected in poland on 32 host plants. in europe it was noted on melittis melisophyllum in austria, france, switzerland and montenegro (braun 1995; farr et al. [n.d.]). phyllactinia mali (duby) u. braun specimen examined: on crataegus sp. (cult): wyżyna lubelska upland, lublin, road side, 5.11.2004, lbl m-8649. remarks. in poland this species was recorded on this host in opole and pruszków (sałata 1985). podosphaera xanthii (castagne) u. braun & n. shishkoff [= sphaerotheca xanthii (cast.) l. junell] white amphigenous mycelium on the leaves, appressoria indistinct, sometimes slightly nipple-shaped. conidia ellipsoid-ovoid: 25-27.5 x 15-17.5 μm produced in chains. the ascocarps grouped; they occurred on both sides of leaves and measured 87-115 (-125) μm in diameter. the appendages on the lower part of ascocarp; they were septate and brown the longest ones were subhyaline at the end. in the chasmothecium one sessile ascus: 60-70 x 50-60 μm. the oculus (apical thin-walled part of ascus) large, measured 15-20 μm in diameter (fig. 2a). ascospores broadly ellipsoid: 13,5-17,5 x 10-14 μm. specimens examined: on physalis alkekengi: western carpathians, pogórze środkowobeskidzkie foothills, rymanów, garden, cultivated, 9.09.2002, lbl m-8651; wyżyna lubelska upland, lublin, garden, cultivated, 19.09.2003, 21.09.2003, 22.10.2003, lbl m-10049–10051; remarks. sphaerotheca fusca (fr.) blumer sensu braun (1987, 1995) was divided into two species: podosphaera xanthii (castagne) u. braun & n. shishkoff and podosphaera fusca (fr.) u. braun & n. shishkoff due to the diameter of oculus and the size of ascomata (braun, takamatsu 2000). the polish collections on physalis alkekengi belong to podosphaera xanthii – the oculi (thin, apical parts of asci) are large and measured 15-20 μm (fig. 2a). for comparison was used specimen published by wołczańska and oklejewicz (2001) as sphaerotheca fusca (fr.) blumer (=podosphaera fusca (fr.) u. braun & n. shishkoff) on doronicum austriacum jacq., beskid niski mts., jasiel, 31.07.1991, lbl m-7126. the oculi of asci of this species measure 10-12 μm and are distinctly smaller than those of podosphaera xanthii (fig. 2b). new records of erysiphales 73 in connection to the diagnosis proposed by braun and takamatsu (2000) other specimens reported as sphaerotheca fusca sensu braun (1987, 1995) require revision. in the mycological literature there is only little information about the occurrence of podosphaera xanthii on physalis spp. it was recorded under this name only in taiwan (cheng et al. 2006) and as sphaerotheca xanthii from bulgaria (farr et al. [n.d.]). uredinales aecidium clematidis dc. specimen examined: on clematis sp. (cult.): western carpathians, pogórze środkowobeskidzkie foothills, rymanów, garden, 24.06.2006, lbl m-8648. remarks. in poland this species was recorded only on clematis recta in kazimierz dolny, puławy (majewski 1977) and in kazimierski landscape park (romaszewskasałata et al. 1991-1992). puccinia mei-mamillata semad. pycnia: hypogenous on leaves, scattered between aecia, diameter 80 um. aecia: aecidium type, as longitudinal or roundish, orange groups up to 6 mm, on leaf petioles and hypogenous on leaves along the nerves. the outer wall of peridium cells distinctly thicker [8-12 μm] than the inner wall [3-6(8) μm]. aeciospores angular globoid, 16 x 20 μm, delicately verrucosus with distinct clots, spore wall hyaline 0.5-1 μm (fig. 1 c). specimen examined: on angelica sylvestris: western carpathians, pogórze środkowobeskidzkie foothills, rymanów, meadow, 21.05.1992, lbl m-8584. remarks. puccinia mei-mamillata is a heteroaecious species. so far in poland aecia of this fungus have been collected only on mutellina purpurea (poir.) thell. on śnieżnik mt (sudety mts) and on babia góra mt (beskid żywiecki mts) (majewski 1979) and lately also in the tatra mountains (wołczańska, piątek 2008, mscr.). urediniospores and teliospores occur on polygonum bistorta l. in the south and in the middle part of poland (majewski 1979) and on polygonum viviparum l. in the tatra national park (beskid) (sałata et al. 1984). on angelica sylvestris this species has not been collected in poland so far. it was recorded on this host in the czech republic, lithuania, norway, germany, russia and sweden (farr et al. [n.d.]). in poland aecia of the following agents of rust fungi can occur on angelica species (majewski 1979): puccinia bistortae dc., puccinia polygoni-vivipari karst., puccinia pimpinellae (str.) röhl. and puccinia mei-mamillata. the aeciospores of puccinia mei-mamillata are the smallest and have big, seceding clots. important differences are also visible in the thickness of peridium cells. p. mei-mamillata has different proportion between the outer and inner peridium walls than puccinia pimpinellae and the thickness of peridium cell walls of the other species is distinctly smaller (tab.1). key to the aecial state of rust fungi on angelica ssp. in poland 1. aeciospores with distinct, big, seceding clots ......................puccinia mei-mamillata 1* aeciospores without clots ......................................................................................... 2 2. the inner wall of peridium cells thicker than the outer wall ..puccinia pimpinellae 2* the inner wall of peridium cells thiner than the outer wall ................................... 3 3. aeciospores big, 20-32 x 14-21 μm; the spore wall 2-3 μm ........... puccinia bistortae 3* aeciospores smaller, diameter 18-25 μm; the spore wall 3-4 μm ............................ .................................................................................................. puccinia polygoni-vivipari 74 a. wołczańska puccinia sessilis schneid. specimen examined: on platanthera bifolia (l.) rich.: roztocze, roztocze national park, maziarki reserve, beech forest, 25.06.1998, lbl m-8647. remarks. in poland this species was recorded on this host only in lipiny near lubin (majewski 1979). uromyces ononidis pass. specimen examined: on ononis arvensis l.: western carpathians, pogórze środkowobeskidzkie foothills, rymanów, waste land, 21.09.1994, lbl m-8646. remarks. in poland this species was recorded on this host near międzyrzec podlaski (majewski 1977) and in rymanów (wołczańska 1994). references adamska i. 2001. microscopic fungus-like organisms and fungi of the słowiński national park (nw poland). ii. acta mycol. 36 (1): 31–65. adamska i. 2005. fungal species new in poland on carex and juncus. acta mycol. 40 (1): 19–24. braun u. 1987. a monograph of the erysiphales (powdery mildews). beihefte zur nova hedwigia 89, berlin-stuttgart. braun u. 1995. the powdery mildews (erysiphales) of europe. gustav fisher verlag, jena stuttgartnew york. braun u. 1999. some critical notes on the classification and generic concept of the erysiphaceae. schlech-schlechtendalia 3: 48–54. braun u., takamatsu s. 2000. phylogeny of erysiphe, microsphaera, uncinula (erysipheae) and cystotheca, podosphaera, sphaerotheca (cystotheceae) inferred from rdna its sequences – some taxonomic consequences. schlechtendalia 4: 1–33. cheng c. w., chen r. s., chang w. h., tsay j. g. 2006. the occurrence of powdery mildew on physalis angulata caused by podosphaera xanthii. plant prot. bull. 48: 41–51. farr d.f., rossman a.y., palm m.e., mccray e.b. [n.d.]. fungal databases, systematic mycology and microbiology laboratory, ars, usda. retrived june 10, 2008, from http://nt.ars-grin.gov/fungaldatabases/. majewski t. 1977. flora polska. grzyby (mycota) 9: basidiomycetes, uredinales. i. pwn, warszawa– kraków. majewski t. 1979. flora polska. grzyby (mycota) 11: basidiomycetes, uredinales. ii. pwn, warszawa– kraków. mirek z., piękoś-mirkowa h., zając a., zając m. 2002. flowering plants and pteridophytes of poland. a checklist. w. szafer institute of botany, polish academy of sciences, kraków. piątek m. 2003 a. erysiphe azaleae and erysiphe syringae-japonicae introduced in poland. mycotaxon 87: 121–126. ta b l e 1 comparison of aecia of rust fungi occurring on angelica spp. in poland species aeciospores peridium dimensions occurrence of clots outer cell inner cell puccinia bistortae* 20-32x14-21 μm – 3-3,5 μm 2-3 μm puccinia polygonivivipari* ø 18-28 μm – to 5 μm thinner than the outer wall puccinia pimpinellae* ø 18-25 μm – 4-5 μm 4-8 μm puccinia meimamillata ø 16-20 μm + 8-12 μm 3-6(-8) μm * data after majewski (1979) new records of erysiphales 75 piątek m. 2003 b. puccinia lagenophorae (urediniomycetes), a neomycete new in poland. polish bot. j. 48 (1): 83–85. piątek m. 2005. first report of powdery mildew of ligustrum caused by erysiphe syringae-japonicae. pl. pathol. 54: 578. piątek, m., wołczańska a. 2004. some phytopathogenic fungi rare or new to poland. polish bot. j. 49 (1): 67–72. romaszewska-sałata j., sałata b., mułenko w. 1991-1992. wstępne uwagi o mikroskopowych grzybach fitopatogenicznych kazimierskiego parku krajobrazowego. folia soc. sci. lubl. 32, biol. (1/2): 31–39. ruszkiewicz-michalska m. 2006. mikroskopijne grzyby pasożytnicze w zbiorowiskach roślinnych wyżyny częstochowskiej. monogr. bot. 96: 1–140. ruszkiewicz-michalska m., michalski m. 2005. puccinia scillae (uredinales) fungus new for poland. acta mycol. 40 (1): 63–70. sałata b. 1985. flora polska. grzyby (mycota) 15: ascomycetes, erysiphales. pwn, warszawa-kraków. sałata b., romaszewska-sałata j., mułenko w. 1984. notatki mikologiczne z tatrzańskiego parku narodowego. acta mycol. 20 (1): 13–21. wołczańska a . 1994. new localities of some rare species of uredinales in poland. acta mycol. 29 (1): 95–98. wołczańska a. 2007. first report of erysiphe carpinicola (perfect state) in poland. pl. pathol. 56 (2): 354. wołczańska a., lamorski t. 2006. puccinia laserpitii (uredinales), a new species for poland. polish bot. j. 51 (2): 221–224. wołczańska a., oklejewicz k. 2001. new and rare species of parasitic fungi in poland. acta mycol. 36 (1): 7–12. wołczańska a., piątek m. 2008. some phytopathogenic fungi rare or new to poland – second contribution (mscr.). wołczańska a., rozwałka r. 2005. urocystis muscaridis (ustilaginomycetes), a fungal species new in poland. polish bot. j. 50 (1): 93–96. nowe informacje o grzybach fitopatogenicznych w polsce s t r e s z c z e n i e w pracy przedstawiono nowe stanowiska dla siedmiu gatunków grzybów fitopatogenicznych. trzy z nich znaleziono na roślinach żywicielskich, na których nie były dotąd notowane w polsce: puccinia mei-mamillata (na angelica sylvestris), neoerysiphe galeopsidis (na melittis melisophyllum) i podosphaera xanthi (na physalis alkekengi). podano dla nich opisy i ilustracje, a dla grzybów rdzawnikowych tworzących ecja na przedstawicielach rodzaju angelica przygotowano klucz do ich oznaczania. pozostałe gatunki na podanych żywicielach znane są z niewielu stanowisk w polsce. fig. 1. a – lobed appressoria of neoerysiphe galeopsidis; b – germinating conidium of neoerysiphe galeopsidis; c – aeciospores of puccinia mei-mamillata on angelica sylvestris, scale bar = 20 μm. a c b fig. 2. a – ascus of podosphaera xanthii; b – ascus of podosphaera fusca, scale bar = 20 μm. a b 2014-01-01t11:47:21+0100 polish botanical society sistotrema luteoviride sp. nov. (cantharellales, basidiomycota) from finland heikki kotiranta1 and karl-henrik larsson2 1finnish environment institute, natural environment centre, biodiversity unit p.o. box 140, fi-00251 helsinki, heikki.kotiranta@ymparisto.fi 2natural history museum, university of oslo p.o. box 1172 blindern, no-0318 oslo, k.h.larsson@nhm.uio.no kotiranta h., larsson k.-h.: sistotrema luteoviride sp. nov. (cantharellales, basidiomycota) from finland. acta mycol. 48(2): 219–225, 2013. a new sistotrema species from northern finland, s. luteoviride is described and illustrated. the two hitherto known collections derive from finnish lapland and both grew on corticated juniperus communis. the spores are very similar to those of s. citriforme, which however is a simple septate species and differs clearly by its its sequence. key words: cantharellales, juniperus communis, lapland introduction sistotrema fr. is a comparatively large genus (index fungorum 2013) typified by the stipitate species s. confluens fr. despite the morphology of the type, all other species presently referred to sistotrema have effused basidiocarps with a smooth, hydnoid or poroid hymenophore. the type species together with a few poroid or hydnoid species probably all have an ectomycorrhizal habit (nilsson et al. 2006; münzenberger et al. 2012) while the majority of species seem to be saprophytes. according to nilsson et al. (2006) the genus is non-monophyletic, and most likely the species outside the core group around the type must be distributed over several genera (larsson 2007). during a survey of corticioid species dwelling on juniperus communis l. in estonia (sell, kotiranta 2011) and finland, two collections of an unknown species with morphological similarities to sistotrema citriforme (m.p. christ.) k.h. larss. & hjortstam were discovered in finnish lapland. molecular phylogenetic analyses confirmed its affinity to sistotrema and its close relationship to s. citriforme. the species is here characterized and described based on morphological and molecular information. acta mycologica vol. 48 (2): 219–225 2013 doi: 10.5586/am.2013.023 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 220 h. kotiranta and k.-h. larsson materials and methods thirty spores per specimen were measured in cotton blue (cb) or melzer´s reagent (iki). cb+ means that the walls of the cells are stained by cotton blue and cb–, that they are not stained, and iki– that there is no reaction to melzer´s reagent. the following abbreviations are used: l* = mean spore length, w* = mean spore width, q = range of the variation in l/w ratio, q* = quotient of the mean spore length (l/w). none of the measurements derive from a spore print. biological province of the collecting site in finland is indicated according to the finnish national uniform grid system (27°e), as applied to biological material by heikinheimo and raatikainen (1981). nine sequences from the nuclear ribosomal its and lsu regions are newly published for this study. data on vouchers and on sequences downloaded from genbank and included in phylogenetic analyses are provided in table 1. dna was extracted from dried specimens (tab. 1) using dneasy plant mini kit (qiagen), following manufacturer’s recommendations. pcr reactions were carried out using ready-to-go™ pcr beads (amersham pharmacia biotech). primers used to amplify the complete its region and the 5´end of the lsu region were its1f and its4b (gardes, bruns 1993), and lr0r and lr7 (hopple, vilgalys 1999) respectively. amplified products were purified using qiaquick spin columns (qiagen). primers used for sequencing were its3, its4 (white et al. 1990), lr5, lr3r (hopple, vilgalys 1999), and ctb6 (http://plantbio.berkeley.edu/bruns/). sequencing was done by macrogen (south korea). sequences were edited and assembled using sequencher 3.1 (gene codes, ann arbor). table 1 species and specimens included in the phylogenetic analysis species origin coll. nr herb. genb. nr clavulina cinerea finland k-h larsson 11694 gb eu118616 c. cristata finland e larsson 6/00 gb membranomyces spurius sweden k hjortstam 19169 gb protodontia piceicola sweden k-h larsson 11803 gb dq873660 sistotrema brinkmannii sweden k-h larsson 14078 gb s. citriforme sweden k-h larsson 15898 gb s. confluens canary islands b. nordén gb am259216 s. coroniferum sweden k-h larsson gb s. luteoviride finland kotiranta 23176 & sell h s. muscicola finland k-h larsson 11721 gb aj606040 s. oblongisporum sweden k-h larsson 14077 gb s. pistilliferum spain e larsson 28/10 gb s. raduloides finland l ryvarden 44004 gb a dataset with 12 sequences representing sistotrema, membranomyces jülich, and clavulina j. schröt. was compiled. protodontia piceicola (bourdot) g.w. martin was added as outgroup. the dataset had 2378 nucleotide positions when the 5.8 and lsu regions had been manually aligned. its1 and 2 were left unaligned and not included in the analysis. the analysed matrix had 1397 characters of which 115 were parsimony informative and 183 variable but uninformative. maximum parsimony was performed using paup* 4.1b10 (swofford). all transformations were sistotrema luteoviride sp. nov. from finland 221 considered unordered and equally weighted. gaps were treated as a fifth character. heuristic searches used 500 random taxon addition replicates and tbr branch swapping with other options using the program’s default settings. relative robustness of clades was estimated through bootstrap analysis using paup*. settings used were 500 bootstrap replicates with 100 random addition sequences per replicate, tbr branch swapping, and otherwise default settings. results the phylogenetic analysis generated 4 most parsimonious trees, one of which is shown as a phylogram in figure 3. this tree had a consistency index 0.7137. the new species clusters with sistotrema citriforme with high bootstrap support (98%). they form together with s. pistilliferum hauerslev, membranomyces spurius (bourdot) jülich, and two clavulina species a moderately supported clade (79%). the ingroup is not resolved as monophyletic and the paraphyletic nature of sistotrema is again indicated (nilsson et al 2006). however, the restricted dataset used in this study was not designed to reveal any new information regarding the phylogeny of sistotrema. the its sequences of sistotrema citriforme and s. luteoviride described below differ by 70-73% depending on alignment method and including the conservative 5.8s gene. a blast search in genbank with the its2 sequence from sistotrema luteoviride recovered three sequences with 100% similarity. two of them were generated from root-tips of salix reticulata l. in an alpine cliff ecosystem in north sweden (ryberg et al. 2009; genbank accession numbers fm202801, fm203027). the third sequence was recovered from roots of evergreen quercus laurina humb. & bonpl. in a tropical cloud forest in mexico (morris et al. 2009; genbank accession number fj196906). sistotrema luteoviride kotir. & k.h. larss., sp. nov. figs 1, 2 mycobank no. mb 804722 fructificatio resupinata, adherens, hymenophorum laeve, reticulato – porosum, luteoviride, systema hypharum monomiticum, hyphis fibulatis, cystidia desunt; basidia urniformia vel subcylindracea, (19–)21–36(–40) × 6–8(–9) μm, 6 (5–7) sterigmatibus, sporae laevae, late ellipsoideae vel pyriformes, (4.6–)5–6 × (3.6–)4–4.8(–5) μm. holotype: finland. inarin lappi (lapponia inarensis): utsjoki, kevo strict nat. reserve, keneskoski, mixed river-side forest with betula pubescens ssp. czerepanovii (orlova) hämet-ahti, salix spp., and juniperus communis l., on fairly hard, corticated juniperus, diameter 3 cm; also tomentella sp., 69°43´n, 27°02´e, alt 84 m a.s.l. (grid 27°e:7740:(3)500), 23 sept. 2009, leg. h. kotiranta 23176 & sell (h). etymology: the epithet luteoviride refers to the colour of the fruit body, which is yellow with a greenish hue. basidiocarp resupinate, relatively thick and tough, adnate, almost smooth, finely porose – reticulate under the lens, yellow, yellowish or yellowish green, margin distinct clear, or thinning out, not differentiated, without rhizomorphs. hyphal system monomitic, hyphae clamped, partly filled with oily contents, in subiculum fairly thinto relatively thick-walled, cb+, either “normal” long-celled, 3–4 μm in diameter (minority), or short-celled, almost isodiametric, 6–9 μm in diam. 222 h. kotiranta and k.-h. larsson subhymenial hyphae thin-walled, (2.5–)3–4(–4.5) μm wide. cystidia none. basidia urniform or subcylindrical, with oily contents, basally clamped, (19–)21–36(–40) × 6–8(–9) μm, with normally 6 (5–7), up to 7 μm long curved sterigmata. spores at first broadly ellipsoid, when fully ripe pyriform – subangular, sometimes almost biapiculate, (4.7–)5–5.6(–5.8) × 3.6–4.6(–5) μm, l* = 5.2 μm, w* = 4.1 μm, q = 1.1–1.5, q* = 1.3 (kotiranta 23176 & sell, type), (4.6–)5–6 × 4–4.8(–5) μm, l* = 5.3 μm, w* = 4.4 μm, q = 1.1–1.5, q* = 1.2 (kotiranta 23126) often with oily contents, with a prominent, up to 1 μm long apiculus, thin-walled, cb–, iki–. additional specimen examined: finland. inarin lappi (lapponia inarensis): utsinarin lappi (lapponia inarensis): utsjoki, kevo strict nat. reserve, close to the biological station, pinus sylvestris l. dominated dry heath forest site, on corticated living juniperus, diameter 4 cm; also hy­ phoderma argillaceum (bres.) donk, tomentella sp., 69°45´n, 27°00´e, alt 105 m a.s.l. (grid 27°e:77417:(3)5008), 20 sept. 2009, leg. h. kotiranta 23126 (h). in the field, the colour of sistotrema luteoviride resembles that of leptosporomyces galzinii (bourdot) jülich, but is more intensively yellow, and the basidiocarp is not soft and pliable but tough and adnate. in microscope the species are quite different. fig. 1. sistotrema luteoviride kotir. & k.-h. larss. (h. kotiranta 23176, type): a – section through basidiocarp showing the hyphal structure, basidia and spores, – b basidia, – c spores. sistotrema luteoviride sp. nov. from finland 223 the new species resembles by its spores sistotrema citriforme (syn. s. subanguli­ sporum k.h. larss. & hjortstam, see eriksson et al. 1984, p. 1365), but the latter is simple septate and the hyphae are much narrower, without isodiametric, wide cells. also the basidia are larger in s. luteoviride. similar spores are also present in s. pis­ tilliferum but the latter species differs by presence of cystidia. all three species have clearly different ribosomal sequences (fig. 3). nilsson et al. (2006) recovered a clavulina clade that also included membrano­ myces delectabile (h.s. jacks.) kotir. & saaren., a corticioid fungus but with cla­ vulina-like basidia. here we included m. spurius, the second species of the genus, with similar result. tedersoo et al. (2003) identified m. delectabile from root-tips in a mixed forest in estonia and clavulina species are generally regarded as forming ectomycorrhiza and most likely all species in the clade are mycorrhizal (tedersoo et al. 2010). here we show for the first time that species with a morphology fitting a traditional sistotrema concept also can occur in the clavulina clade. the position of s. luteoviride, s. citriforme, and s. pistilliferum among species of clavulina indicates a mycorrhizal habit, which is also underlined by the environmental sequences that we found in genbank. even if there are hitherto only two specimens of s. luteoviride, we believe that it is not an especially rare species in the subarctic areas of the northern hemisphere. the habitats, riveror brook-side forests, nutritionally poor pinus sylvestris dominated forests, and alpine heath vegetation, are very common in the north. the substrate in both collections was a corticated juniperus communis, but if we believe that the species is a symbiont common juniper is unlikely to be the host. rather the juniper fig. 2. sistotrema luteoviride kotir. & k.-h. larss. (h. kotiranta 23126): a – almost isodiametric hyphae in subiculum, – b basidia at different stages of development, – c spores. 224 h. kotiranta and k.-h. larsson bushes happened to offer a suitable substrate where s. luteoviride could develop its resupinate, effused basidiocarps. acknowledgements. we are grateful to teuvo ahti (helsinki) who helped us with the latin diagnosis. ellen larsson, university of gothenburg, is acknowledged for her proficiency in producing the dna sequences. the senior author thanks anton schiryaev (ekaterinburg) and indrek sell (tartu) for the nice companionship during the collection trip in lapland. also the staff of kevo biological station is warmly thanked for the hospitality during our stay at the station. references eriksson j., hjortstam k., ryvarden l. 1984. the corticiaceae of north europe. schizopora – suillo­ sporium. 7: 1281-1449. gardes m., bruns t.d. 1993. its primers with enhanced specificity for basidiomycetes--application to the identification of mycorrhizae and rusts. mol. ecol. 2 (2):113-118. http://www.ncbi.nlm.nih.gov/ pubmed/8180733 heikinheimo o., raatikainen m. 1981. ruutukoordinaattien ja paikannimien käyttö suomessa [grid references and names of localities in the recording of biological finds in finland]. notul. entomol. 61: 133-154, [in finnish]. hopple j.s.jr., vilgalys r. 1999. phylogenetic relationships in the mushroom genus coprinus and dark-spored allies based on sequence data from the nuclear gene coding for the large ribosomal subunit rna: divergent domains, outgroups, and monophyly. molecular phylogenetics and evolution 13 (1):1-19. index fungorum. 2013. sistotrema. www.indexfungorum.org/names/names.asp, 15.3.2013. larsson k.-h. 2007. re-thinking the classification of corticioid fungi. mycological research 111: 1040–1063. morris m.h., pérez-pérez m.a., smith m.e., bledsoe c.s. 2009. influence of host species on ectomycorrhizal communities associated with two co-occurring oaks (quercus spp.) in a tropical cloud forest. fems microbiology ecology 69: 274-287. münzenberger b., schneider b., nilsson r.h., bubner b., larsson k.-h., hüttl r.f. 2012. morphology, anatomy, and molecular studies of the ectomycorrhiza formed axenically by the fungus sistotrema sp. (basidiomycota). mycological progress 11: 817-826. fig. 3. phylogram of one of four trees of length 468 from the maximum parsimony analysis. numbers on branches show all bootstrap values higher than 75 %. the scale indicates number of changes along the branches. sistotrema luteoviride sp. nov. from finland 225 nilsson r.h., larsson k.-h., larsson e., kõljalg u. 2006. fruiting body-guided molecular identification of root-tip mantle mycelia provides strong indications of ectomycorrhizal associations in two species of sistotrema (basidiomycota). mycological research 110: 1426-1432. ryberg m., larsson e., molau u. 2009. ectomycorrhizal diversity on dryas octopetala and salix reticulata in an alpine cliff ecosystem. arctic, antarctic, and alpine research 41: 506-514. http://dx.doi.org/1 0.1657%2f1938-4246-41.4.506 sell i., kotiranta h. 2011. diversity and distribution of aphyllophoroid fungi growing on common juniper (juniperus communis l.) in estonia. folia cryptogamica estonica 48: 73-84. tedersoo l., kõljalg u., hallenberg n., larsson k.-h. 2003. fine scale distribution of ectomycorrhizal fungi and roots across substrate layers including coarse woody debris in a mixed forest. new phytologist 159: 153-165. http://dx.doi.org/10.1046%2fj.1469-8137.2003.00792.x tedersoo l., may t.w., smith m.e. 2010. ectomycorrhizal lifestyle in fungi: global diversity, distribution, and evolution of phylogenetic lineages. mycorrhiza 20: 217-263. http://dx.doi.org/10.1007%2fs00572009-0274-x 2013-12-20t14:47:11+0100 polish botanical society therrya fuckelii and other fungi on stems and branches of pinus sylvestris following lightning damage hanna kwaśna and piotr łakomy department of forest pathology, poznań university of life science wojska polskiego 71c, pl-60-625 poznań, kwasna@up.poznan.pl kwaśna h., łakomy p.: therrya fuckelii and other fungi on stems and branches of pinus sylvestris following lightning damage. acta mycol. 46 (1): 109–114, 2011. the evidence-based hypothesis is presented that the stems and branches of pinus sylvestris injured by lightning strikes are colonized first by therrya fuckelii and successively by diplodia pinea, nectria fuckeliana, hyaloscypha leuconica, gremmeniella abietina and cenangium ferruginosum. the concomitant occurrence of these usually pathogenic fungi on injured pinus trees in poland signals a potential for their increased significance in europe during climatic changes. key words: pathogenic fungi, lightning damage, climatic changes introduction scots pine (pinus sylvestris l.) is amongst the most common trees occurring throughout the hemiboreal forests. it is an important tree in polish commercial forestry. it is subject to a number of damaging biotic and abiotic factors. in the last few years, european meteorological data show a steady increase in the incidence and violence of storms and winds, accompanied by heavy rain, hail and lightning in the highlands and lowlands (anonymous 2005). these changes increase the incidence of mechanical damage to trees and, as a consequence, susceptibility to pathogenic fungi. the discovery of several groups of dying trees in 50-100-year-old scots pine stands in northwest poland in the summer of 2006, with no etiological symptoms of crown or butt and root rot pathogens, or pest infestation, prompted a closer investigation followed by morphological examination of fungi occurring on stems and branches. here we report the results of morphological analyses of ascomycota occurring on dying scots pines which have been damaged by electrical bursts during thunderstorms. acta mycologica vol. 46 (1): 109–114 2011 110 h. kwaśna and p. łakomy materials and methods samples were collected from groups of dying 50-100-year-old p. sylvestris trees located in babki (52o23’n, 16o58’e), skwierzyna (52o38’n, 15o31’e) and tuczno (53o16’n, 16o11’e) forest districts in northwest poland in july 2006. four pieces of stems (50 cm in length x 10-15 cm diam.) and ten branches with twigs (50 cm in length x 0.55 cm diam.) were collected from each; i.e. trees and from the litter in each focus. in total 12 stems and 30 branches from each: trees and the litter, were collected in three forest districts. branches were with or without needles attached, often with yellowish-green and brittle bark, with black apothecial initials on the bark surfaces, occasionally with necrosis and bark canker. observation of other symptoms of diseases on analysed trees continued until late november 2006. samples were incubated in moist chambers at 25oc for 8 months. each month, fungal apothecia and pycnidia appearing on the bark surface were examined macro and microscopically; in water, in 0.1% cotton blue in water, and in melzer’s reagent (3% potassium iodide + 1% iodine, in water + chloral hydrate, 1:1, v:v). fungi were isolated by placing non-disinfected fragments of hymenium, asci, ascospores or conidia on pda (filtered white potatoes 40 g, glucose 20 g, agar 20 g, distilled water 1 l, ph = 7 or ph = 4), ma (malt extract 30 g, agar 15 g, distilled water 1 l), mea (malt extract 30 g, peptone from soymeal 3 g, agar 15 g, distilled water 1 l) and sna (kh2po4 1 g, kno3 1 g, mgso4·7h2o 0.5 g, kcl 0.5 g, glucose 0.2 g, sucrose 0.2 g, agar 20 g, distilled water 1 l). fungi were identified on the basis of colony morphology and sporulation. the incidence of individual fungi (%) was calculated as percentage occurrence in samples examined in the laboratory. the statistical significance of differences in numbers of stems or branches colonized by particular fungal species in two locations was determined by χ2 test. results circular, 30-50 m wide areas, each including 10-32 dying scots pines were observed in the babki, skwierzyna and tuczno forest districts, in northwest poland, in early summer 2006 (fig. 1). the affected trees had 10-50% branches and twigs dead, still attached or gradually abscising. the whole of dead upper parts of the main stems (4-6 m in length) and dying branches were covered with scattered or clustered black apothecia. the bark was intact or brittle and discolored yellowish-green (figs 2, 3). there were more apothecia on stems with intact bark and fewer on stems with brittle and discolored bark. needles had been dying and falling continuously for 3-4 months from the infected but usually attached branches. the death of trees progressed and spread to trees located around the initially affected area. adjacent trees showed first symptoms of decline within 4-5 months. only one in 200 primarily and secondarily affected trees had an elongated, 5 m long wound that could have been caused by a lightning strike. therrya fuckelii 111 after 20 days of incubation in moist chambers, the black apothecia opened and revealed asci and ascospores typical of therrya fuckelii (rehm) kujala (minter 1996). asci matured sequentially, were clavate, with a flat apex, (70-) 105-160 (-191) x (6.5-) 10-15 (-17) μm. ascospores were long, thin, vermiform to cylindrical, with delicate thread-like tips, parallel, hyaline, thin-walled, smooth, (64-) 65-110 (-157) x (2.5-) 3-4 (-4.5) μm (figs 4-6). the necrotic phloem and partly necrotic xylem occurred on stems colonized by t. fuckelii. no further fungi or pests occurred on stems and branches covered densely with t. fuckelii apothecia. between 35-60% of stems and branches moderately covered with t. fuckelii apothecia showed additional sporulation by diplodia pinea (the ‘type a’ of sphaeropsis sapinea (fr.: fr.) dyko & sutton) (the cause of the sphaeropsis tip blight) (tab. 1). between 0-25% of stems and branches showed sporulation by nectria fuckeliana (the cause of the stem canker) and hyaloscypha leuconica (usual colonizer of the dead tissues). between 0-50% of stems and branches from upper parts of diseased crowns showed sporulation by gremmeniella abietina var. abietina (the ‘type a’ of the european race, the cause of the scleroderris canker) and cenangium ferruginosum (the cause of the cenangium canker). the differences in number of stems or branches colonized by particular fungal species in two different locations were often statistically significant. sporulation by secondary invaders appeared on stems and branches in the forest and in the laboratory, 4-8 months after t. fuckelii apothecia were first noted. the ‘type a’ of s. sapinea was determined on the basis of size of conidia (palmer et al. 1987). the ‘type a’ of the european race of g. abietina was determined on the basis of symptom expression of colonized trees in field and growth properties, production of conidia in vitro, septation of conidia (hellgren 1995; hellgren, högberg 1995). only occasional samples colonized by d. pinea showed symptoms of initial advanced bark necrosis. table 1 the percentage of scots pine stems and branches moderately colonized by t. fuckelii invaded by secondary colonizers in babki, skwierzyna and tuczno forest districts secondary colonizer babki skwierzyna tuczno stems branches stems branches stems branches cenangium ferruginosum fr. 10 35ab 10 15a 10 10b diplodia pinea (desm.) j. kickx f. 40 50 50 35c 40 60c gremmeniella abietina (lagerb.) m. morelet 0 50b 0 40d 0 10bd hyaloscypha leuconica cooke) nannf. 0 10e 0 5c 0 0ec nectria fuckeliana c. booth 10ae 15ab 0a 25ad 0e 0bd abbreviations: a – the ratio for stems or branches in babki and skwierzyna is significantly different from 1:1 at p = 0.05; b – the ratio for stems or branches in babki and tuczno is significantly different from 1:1 at p = 0.001; c – the ratio for stems or branches in skwierzyna and tuczno is significantly different from 1:1 at p = 0.05; d – the ratio for stems or branches in skwierzyna and tuczno is significantly different from 1:1 at p = 0.001; e – the ratio for stems or branches in babki and tuczno is significantly different from 1:1 at p = 0.05. 112 h. kwaśna and p. łakomy discussion therrya fuckelii was the first fungus observed on upper stems and branches of dying scots pine trees grouped in approximately circular 30-50 m diam. foci, in three forest districts situated 100-150 km apart in northwest poland, in 2006. apothecia of the fungus were present along the whole length of infected branches. no other primary pathogens of scots pine stem tops, e.g. cronartium flaccidum (alb. & schwein.) g. winter and endocronartium pini (willd.) y. hirats. were observed. minter (1996) and torp (2004) reported that t. fuckelii was commonly found on freshly killed, dead or self-pruned, 0.5-7 cm diam. pinus branches and twigs, either attached to the tree (mostly in lower parts of the crown) or fallen. the bark on infected branches was brittle and discolored bright red to brown. unlike fallen and non-colonized twigs, the stems and branches colonized by t. fuckelii never had needles attached. a role for t. fuckelii in self-pruning of scots pine has been suggested (minter 1996). torp (2004) found t. fuckelii in the snag tops of injured p. sylvestris trees in norway, in 2003-2004. the provenance of six specimens of the fungus deposited in the norwegian herbarium prompted torp (2004) to conclude that t. fuckelii occurs in a very specialized niche. after careful examination of the snag tops of the injured trees torp (2004) concluded that colonization by t. fuckelii may have been preceded by damage from electrical bursts during thunderstorms. the occurrence of t. fuckelii in poland followed severe weather in 2004-2005, when the northwest part of the country had 180 days with rainfall, 20 days with thunderstorms, more than 40 days with severe frosts, relatively high humidity (78-84%) and strong winds (4 m s-1 mean). some thunderstorms were particularly severe, with strong wind, hail, flash flooding and abundant lightning. the number of electrical ground strikes in may-september peaked at 12 per km2 in 2004 and 6-20 per km2 in 2005, with the greatest concentration in tuczno. very warm weather preceded the most severe conditions, which included hail storms (anonymous 2005). these observations and those of minter (1996) and torp (2004) suggest that t. fuckelii may specialize in invading the freshly damaged bark. the endophytic status of t. fuckelii cannot be excluded. since d. pinea, n. fuckeliana, h. leuconica, g. abietina and c. ferruginosum, appeared on the affected branches in a succession, these fungi seem to be secondary invaders of stems and branches colonized primarily by t. fuckelii. as the secondary invaders did not colonize the branches strongly colonized by t. fuckelii, the latter fungus may act as a natural antagonist. diplodia pinea, n. fuckeliana, g. abietina and c. ferruginosum have been recognized as endophytes, and weak or latent pathogens of conifers, active only locally and periodically (in specific weather conditions), able to colonize stressed, weakened, injured or dying woody tissues. currently, however, the significance of these fungi in poland is increasing (mańka 2005). diplodia pinea has emerged as a serious problem in the last 10 years (łakomy, pers. comm.). pathogen appears mostly in scots pine plantations, in central and western poland. according to zwolinski et al. (1990) heavy damage of trees and epidemics are usually associated with several years of drought, and frosts, cold and wet therrya fuckelii 113 springs as well as hail damage. diplodia pinea is more aggressive than d. scrobiculata j. de wet, slippers & m. j. wingf. (the ‘type b’ of s. sapinea). pathogenic activity of n. fuckeliana has hitherto been reported only on abies concolor (gord. & glend.) lindl. ex hildebr. and picea abies (l.) h. karst. so far, the fungus has been exceptional on pinus. earlier, only crane (2005) observed n. fuckeliana on p. radiata d. don (in south island of new zealand), in association with the pine fluting disease. gremmeniella abietina and c. ferruginosum periodically cause severe losses of green foliage, mostly in younger (g. abietina) scots pine stands in poland (mańka 2005). it is the first report on the occurrence of these fungi in association with storm and lightning damage. the association between t. fuckelii and d. pinea, n. fuckeliana, g. abietina, c. ferruginosum results from similar preferences for pinus tissues and the weather and climatic conditions. the similar association between therrya piceae funk and lachnellula agassizii (berk. & m.a. curtis) dennis, sarea difformis (fr.) fr., corniculariella abietis p. karst., botryosphaeria piceae a. funk and tryblidiopsis pinastri (pers.) p. karst. was observed in british columbia (canada) on picea glauca (moench) voss where the above mentioned species contributed to the perennial stem cankers (funk 1982). similarly to our observations, stem cankers caused serious damages to spruce but only on the localized small areas. damages acted as indicators of unfavourable growing conditions for spruce. the mineral deficiencies in the soil were considered to be the main predisposing cause (funk 1965). conclusion it is hypothesized that the successive colonization and co-occurrence of t. fuckelii, and d. pinea, n. fuckeliana, g. abietina, c. ferruginosum on scots pine was an ecological response to the changes caused by a physical stimulus, i.e. a lightning strike. the detection of t. fuckelii on the directly or indirectly injured p. sylvestris trees establishes environmental event and may be a powerful predictor of ecological behaviour of other fungal colonizers. the occurrence of t. fuckelii may signal risk from g. abietina, c. ferruginosum and d. pinea with no immediate production of disease in the case of latent infections. the successive colonization of injured scots pines has significant implications for disease management. 114 h. kwaśna and p. łakomy references anonymous 2005. bulletin of the national hydrological and meteorological service. instytut meteorologii i gospodarki wodnej. warszawa. crane p.e. 2005. nectria fuckeliana disease of pinus radiata ecology and epidemiology. report no 38080. www. fbrc.org.nz. funk w. 1965. a new parasite of spruce from british columbia. can. j. plant pathol. 43: 45–48. funk w. 1982. therrya canker of spruce in british columbia. can. j. plant pathol. 4: 357–361. hellgren m. 1995. gremmeniella abietina biology and genetic variation within fennoscandia. doctoral thesis. swedish university of agricultural sciences, department of forest mycology and pathology, uppsala, pp 1–60. hellgren m., högberg n. 1995. ecotypic variation of gremmeniella abietina in northern europe—disease patterns reflected by dna variation. can. j. bot. 73: 1531–1539. mańka k. 2005. fitopatologia leśna. państwowe wydawnictwo rolnicze i leśne. warszawa. pp 391. minter d.w. 1996. imi descriptions of fungi and bacteria. no. 1297 mycopathologia 136: 171–173. palmer m.a., stewart e.l., wingfield m.j. 1987. variation among isolates of sphaeropsis sapinea in the north central united states. phytopathology 77: 944–948. torp t. b. 2004. occurrence of therrya fuckelii (rehm) kujala on scots pines in the eastern part of norway. master degree thesis. norges landbrukshøgskole. zwolinski j.b., swart w.j., wingfield m.j. 1990. economic impact of the post-hail outbreak of dieback induced by sphaeropsis sapinea. eur. j. for. pathol. 20: 405–411. therrya fuckelii i inne grzyby na strzałach i gałęziach pinus sylvestris uszkodzonych przez uderzenia pioruna streszczenie praca stara się udowodnić, że strzały i gałęzie sosny zwyczajnej uszkodzonej uderzeniami pioruna kolonizowane są przez therrya fuckelii, a następnie przez diplodia pinea, nectria fuckeliana, hyaloscypha leuconica, gremmeniella abietina i cenangium ferruginosum. sukcesja i wspólne występowanie tych, w większości patogenicznych, grzybów na uszkodzonych sosnach sygnalizuje wzrost ich potencjału i znaczenia w polsce i europie. wynikają one z obserwowanych ostatnio zmian klimatycznych i występowania pogody obfitującej w częste burze i wyładowania atmosferyczne. 2014-01-01t11:52:05+0100 polish botanical society fungi isolated from phyllosphere of fodder galega (galega orientalis) bożena cwalina-ambroziak1 and stanisław sienkiewicz2 1department of phytopathology and entomology, university of warmia and mazury prawocheńskiego 17, pl-10-722 olsztyn, bambr@uwm.edu.pl 2department of agriculture chemistry and environment protection, university of warmia and mazury oczapowskiego 8, pl-10-722 olsztyn cwalina-ambroziak b., sienkiewicz s.: fungi isolated from phyllosphere of fodder galega (galega orientalis). acta mycol. 43 (2): 173–179, 2008. the object of the experiment was fodder galega (galega orientalis lam.) cultivated in 2001-2003 as field crop on three plots: 1. without fertilization, 2. 40 kg p2o5 × ha -1 and 80 kg k2o × ha -1, 3. 80 kg p2o5 -1 × ha and 160 kg k2o × ha -1. during the dry and warm vegetation season of 2002 almost two times fewer isolates were obtained from the leaves than in 2003 that was the most abundant in fungi. yeasts-like fungi (30% of the total number of isolates) and saprotrophic fungi with dominated species: acremonium strictum (8.5%), genus epicoccum (7.8%), humicola (9.5%) and penicillium (18.9%) were the fungi most frequently populating the leaves of galega. the share of pathogens in the total number of isolates obtained from the phyllosphere was 10.6%. they were represented by fungi of ascochyta spp., botrytis cinerea, genus fusarium, phoma medicaginis and sclerotinia sclerotiorum. reduction by 1.9 to 4.6% in the number of fungi isolated from the phyllosphere of galega without fertilization as compared to galega cultivated in combinations with fertilization was recorded. generally, the smallest number of pathogens was recovered from galega fertilized with 40 kg p2o5 × ha -1 and 80 kg k2o × ha -1. b. cinerea most frequently populated galega in combination without fertilization, genus fusarium fungi in combination without fertilization and with fertilization with 80 kg p2o5 -1 × ha and 160 kg k2o × ha -1, while ascochyta spp. were isolated from galega with fertilization only. key words: pathogenic fungi, saprotrophic fungi, phyllosphere of fodder galega, mineral fertilization acta mycologica vol. 43 (2): 173–179 2008 174 b. cwalina-ambroziak and s. sienkiewicz introduction fodder galega, a perennial legume plant with fine seeds, is relatively immune to viral infections as we learn from reports (kegler, spaar 1996; valkonnen 1993). among fungal pathogens infesting this plant, similar to fabaceae belonging to the same family, alfalfa and clover, botrytis cinerea as well as species of ascochyta and fusarium should be mentioned (cwalina-ambroziak et al. 1999; gorsen et al. 1994; leath, hower 1993; leath et al. 1994). the above-indicated species of fungi were isolated from the phyllosphere of galega by cwalina-ambroziak and koc (2005); they populated galega cultivated as single crop more frequently than galega cultivated in a mix with brome grass. mineral fertilization can also influence the composition of fungal community, which was proven during studies on mycological assessment of rhizosphere, rhizoplane and roots of plants belonging to the above-indicated family (cwalina-ambroziak, majchrzak 2000; deb, bora 1996). the laboratory studies conducted aimed at analysis of the fungal community populating the phyllosphere of fodder galega cultivated under conditions of diversified mineral fertilization. materials and methods fodder galega was cultivated in 2001-2003 at the agricultural experimental enterprise in bałcyny as crop field on good wheat complex soil on three plots of 1 ha each: 1. without fertilization 2. 40 kg p2o5 × ha -1 and 80 kg k2o × ha -1 (17.46 kg p × ha-1 and 66.45 kg k × ha-1) 3. 80 kg p2o5 -1 × ha and 160 kg k2o × ha -1 (34.92 kg p × ha-1 and 132.90 kg k × ha-1). fertilization with p2o5 (superphosphate) and k2o (potassium salt) was applied before sowing.during fl owering of galega samples of 20 leaves were collect-during flowering of galega samples of 20 leaves were collected from plants growing on individual plots from three locations selected at random. at the laboratory fungi were isolated from the collected plant material according to the methodology by chruściak (1974). from the basal part of the leave a fragment of 1 cm2 of leaf blade was cut out and shaken in flasks filled with 200 ml of sterile water. from the suspension of microorganisms prepared in this way 0.2 ml was transferred into petri dishes and covered with glucose and potato medium pda with bengal rose and streptomycin. the fungi growing on the pda medium were transferred after 5 days of incubation at 22°c on pda slants for identification of species according to the keys (arx 1970; booth 1971; ellis 1971; nelson et al. 1983; skirgiełło et al. 1979). colonies of yeasts-like fungi were counted. analysis of variance using the t-duncan test was carried out to determine the influence of mineral reutilization and sampling time on the numbers of fungi most often isolated from leaves of galega (statistica® 6 2001 software package). fungal colonies isolated from phyllosphere 175 results and discussion as a result of three-year studies 1289 fungal isolates among which yeasts-like fungi dominated (30%) were recovered from leaves of fodder galega. the other fungi were represented by 19 species and nonsporulating cultures. saprotrophic fungi were recovered from leaves in large numbers among which genus penicillium species were the most frequent (tab. 1). the other identified fungal species were of genus humicola (9.5%), and acremonium strictum (8.5%), epicoccum purpurascens (7.8%), alternaria alternata (4.2%) and cladosporium cladosporioides (3.5%). chruściak (1974), madej (1997) and wozniakowskaja (1962) classify those fungi as microorganisms commonly populating the leaves of many plant species. additionally those authors inform that yeasts-like fungi populate young leaves of plants first. in this study pathogenic fungi botrytis cinerea (4.9%), genus fusarium (3.2%), ascochyta (2.1%), phoma medicaginis (0.3%) and sclerotinia sclerotiorum (0.5%) were recovered in lower numbers than saprotrophic fungi. three fusarium species were identified: f. avenaceum, f. culmorum and f. equiseti. those pathogens are considered in literature (gorsen et al. 1994; leath et al. 1994) the major causes of diseases in fine seed legumes such as clover and alfalfa. the structure of fungal community in the phyllosphere of fodder galega formed under the influence of application of mineral fertilizers and weather conditions during the consecutive years of study. the largest numbers of fungi were recovered from the phyllosphere of fodder galega in the combination without fertilization and ta b l e 1 fungi isolated from phyllosphere of fodder galega (% of isolates) species 2001 2002 2003 k 40 p 80 p k 40 p 80 p k 40 p 80 p acremonium strictum w. gams 15.1 11.1 4.4 30.3 5.3 7.8 1.5 1.9 alternaria alternata (fr.) keissler 2.0 0.7 0.7 6.3 3.9 9.1 8.4 5.0 ascochyta spp. 2.2 5.8 4.2 4.9 2.0 1.9 botryodiplodia spp. 3.2 botrytis cinerea pers. 7.9 3.7 4.4 2.0 4.2 1.0 9.1 3.9 6.3 cladosporium cladosporioides (fres.) de vries 7.2 5.2 6.6 3.3 3.2 5.9 0.7 1.5 epicoccum purpurascens 5.3 10.4 8.8 4.6 19.0 9.8 5.2 5.9 6.9 fusarium avenaceum (fr.) sacc. 1.3 0.7 7.2 1.1 12.8 0.7 fusarium culmorum (w.g.sm.) sacc. 1.5 1.3 3.2 0.7 fusarium equiseti (corda) sacc. 1.1 2.9 helminthosporium sativum pammel 0.7 humicola fuscoatra traaen 7.2 7.4 3.7 1.3 2.0 0.7 6.4 5.0 humicola grisea traaen 3.0 4.4 7.9 3.2 6.9 10.3 10.8 mortierella alpina peyronel 0.7 0.7 5.3 1.3 mucor circinelloides van tieghem 1.5 1.3 2.0 0.6 mucor hiemalis wehmer 0.7 0.7 5.8 1.5 2.5 penicillium spp. 11.2 23.7 17.5 9.2 19.0 20.6 18.1 31.0 16.4 phoma medicaginis malbr. et roum. 0.7 3.2 sclerotinia sclerotiorum (lib.) de bary 1.0 2.5 non sporulating fungi 3.3 11.1 6.6 0.7 4.2 4.9 3.9 1.5 1.3 yeast-like fungi 38.2 18.5 35.8 30.9 14.7 16.7 33.7 22.6 48.4 total (number of isolates) 152 135 137 152 95 102 154 203 159 explanatations: k – control, 40 p – 40 kg p2o5 × ha -1 and 80 kg k2o × ha -1, 80 p – 80 kg p2o5 × ha -1 and 160 kg k2o × ha -1. 176 b. cwalina-ambroziak and s. sienkiewicz the smallest from combination with fertilization with 80 kg p2o5 × ha -1 and 160 kg k2o × ha -1. however, pathogens were the least frequently isolated from the combination with fertilization with 40 kg p2o5 × ha -1 and 80 kg k2o × ha -1. fusarium spp. and botrytis cinerea populated leaves of galega cultivated on all plots (fig. 1a, b, c). b. cinerea populated galega in combination without fertilization most frequently (6.3%), with a significant difference in numbers as compared to both combinations with fertilization (tab. 2). fungi responsible for wilting of plants colonized the leaves of galega to a similar extent in both plots without fertilization and the one with the highest fertilization rate. fungi of genus ascochyta were not recovered from plants cultivated without fertilization while their share in the total number of fungi recovered from combinations with fertilization was 2.5% (lower fertilization) and 4.0% (higher fertilization). at the same time a larger number of saprotrophic fungi of genera epicoccum and penicillium as well as order mucorales was obtained from plants in combinations with mineral fertilization – 39.5% of the total number of isolates (40 kg p2o5 × ha -1 and 80 kg k2o × ha -1) and 28.4% (80 kg p2o5 × ha -1 and 160 kg k2o × ha -1) as compared to the control combination – 20.7% of isolates. some authors indicate the stimulating influence of mineral fertilization on development of saprotrophic fungi in communities of soil fungi of galega (cwalina-ambroziak, majchrzak 2000) and other fabaceae crops (deb, bora 1996). at the same time a larger number of saprotrophic fungi of genera epicoccum and penicillium as well as order mucorales was obtained from plants in combinations with mineral fertilization – 39.5% of the total number of isolates (40 kg p2o5 × ha -1 and 80 kg k2o × ha -1) and 28.4% (80 kg p2o5 × ha -1 and 160 kg k2o × ha -1) as ta b l e 2 most isolated fungi from phyllosphere of fodder galega (number of isolates) during investigation period species mean for: 2001 2002 2003 acremonium strictum years 14.67 a 19.67 a 2.00 b combination k – 23.00 a 40 p – 7.67 b 80 p – 5.67 b alternaria alternata years 1.33 b 3.67 b 13.00 a combination k – 6.00 b 40 p – 8.00 a 80 p – 4.00 c botrytis cinerea years 7.67 ab 2.67 c 10.67 a combination k – 9.67 a 40 p – 5.67 b 80 p – 5.67 b epicoccum spp. years 11.33 a 11.67 a 10.33 a combination k – 7.67 c 40 p – 14.67 a 80 p – 11.00 ab fusarium spp. years 1.67 b 11.33 a 0.67 b combination k – 5.67 a 40 p – 2.33 b 80 p – 5.67 a humicola spp. years 12.00 bc 8.67 c 20.00 a combination k – 14.00 ab 40 p – 17.33 a 80 p – 9.33 bc penicillium spp. years 24.33 b 17.67 c 39.00 a combination k – 19.67 b 40 p – 37.67 a 80 p – 23.67 b explanations as in table 1. 178 b. cwalina-ambroziak and s. sienkiewicz compared to the control combination – 20.7% of isolates. some authors indicate the stimulating influence of mineral fertilization on development of saprotrophic fungi in communities of soil fungi of galega (cwalina-ambroziak, majchrzak 2000) and other fabaceae crops (deb, bora 1996). analyzing the fungal communities of the phyllosphere of fodder galega during individual vegetation seasons the largest number of isolates were recovered in 2001 and 2003 – 32.9 and 40% of the total number of colonies respectively. b. cinerea was among the most frequently recovered pathogens during those years. gorsen et al. (1994) consider vegetation seasons characterized by high precipitations and moderate temperatures as favorable for the causing agent of grey mould and such conditions existed during the above seasons of study. the lowest number of fungi was recovered from leaves during hot and dry summer of 2002 (27.1%). among the pathogens recovered during that season fungi of genus fusarium dominated and their number was significantly higher than during the other two years of study. conclusions 1. mineral ferilization reduced the total number of fungi colonizing the phyllosphere of fodder galega. 2. ferilization had a varied effect on the development of pathogens. the largest numbers of b. cinerea were recovered from galega in treatment without ferilization, in contrast to the genus fusarium and ascochyta. references arx von j. a. 1970. the genera of fungi sporulating in pure culture. verlag von j. cramer. booth t. c.1971. the genus fusarium. commonwealth mycological institute kew surrey, england. chruściak e. 1974. mikoflora fyllosfery. acta mycol. 10 (1): 173–180. cwalina-ambroziak b., czajka w., wojnowska t. 1999. research on the health state of goats rue (galega orientalis lam.). acta acad. agricult. techn. olst. natur. sc., 2: 17–26. cwalina-ambroziak b., koc j. 2005. grzyby zasiedlające nadziemne organy roślin rutwicy wschodniej (galega orientalis lam.) uprawianej w siewie czystym i w mieszance ze stokłosą bezostną (bromus inermis leyss.). acta agrobot. 58 (1): 125–133. cwalina-ambroziak b., majchrzak b. 2000. the structure of fungal population from galega orientalis root system formed as the result of fertilization. acta mycol. 35 (2): 311–321. deb b., bora k. n. 1996. effect of chemical fertilizer on the rhizosphere mycoflora and nodulation of pea plant. environ. ecol. 14 (4): 747–751. ellis m. b. 1971. dematiaceous, hyphomycetes. cmi, kew, surrey. gorsen b. d. smith s. r., platford r. g. 1994. botrytis cinerea blossom blight of alfalfa on the canadian prines. pl. dis. 78 (12): 1218. kegler h., spaar d. 1996. on the virus susceptibility of galega orientalis lam. arch. phytopath. plant protect. 30 (3): 187–190. leath k. t., de gregorio r. e., ashley r. a. 1994. foliar blight of bigflower vetch caused by ascochyta fabae f. sp. vicia. pl. dis. 78 (6): 637–639. leath k. t., hower a. a. 1993. interaction of fusarium oxysporum f. sp. medicaginis with feeding activity of flover root curculio larvae in alfalfa. pl. dis. 77 (8): 799–802. madej t. 1997. grzyby następczo zasiedlające liście ziemniaka. ochr. rośl. 11: 6–7. nelson p. e., toussoun t.a., marasas w. f. o. 1983. fusarium species. the pensylvania state university press, university park, london. skirgiełlo a., zadara m., ławrynowicz m. 1979. flora polska. grzyby (mycota). 10: glonowce (phycomycetes). pleśniakowe (mucorales). instytut botaniki pan, warszawa-kraków. fungal colonies isolated from phyllosphere 179 valkonnen j. p. t. 1993. resistance to six viruses in the legume goat’s rue (galega orientalis lam.). ann. appl. biol. 123 (2): 309–314. wozniakowskaja j. m. 1962. epiphytic yeast-organism. mikrobiologia 31: 616–622. grzyby wyizolowane z fyllosfery rutwicy wschodniej (galega orientalis) s t r e s z c z e n i e w doświadczeniu poletkowym w bałcynach uprawiano rutwicę wschodnią w następujących obiektach: 1. bez nawożenia, 2. 40 kg p2o5 × ha -1 i 80 kg k2o × ha -1, 3. 80 kg p2o5 -1 × ha i 160 kg k2o × ha -1. założeniem przeprowadzonych badań w aspekcie fitopatologicznym było określenie zbiorowiska grzybów zasiedlających fyllosferę roślin. w tym celu w okresie przed kwitnieniem rutwicy do laboratorium pobierano próby zbiorcze liści z roślin w poszczególnych kombinacjach. izolacje grzybów przeprowadzono zgodnie z metodyką chruściak (1974). liście rutwicy wschodniej były zasiedlone przez 1289 izolatów grzybów reprezentowanych przez 19 gatunków oraz przez grzyby drożdżopodobne i kultury niezarodnikujące. najliczniej wyosobniono grzyby w 2003 roku (616 izolatów), a najmniej licznie w 2002 roku (349). największy udział wśród ogółu izolatów miały grzyby drożdżopodobne (30% ogółu izolatów), mniejszy grzyby saprotroficzne z rodzajów: penicillium (18.9%), humicola (9.5%), epicoccum (7.8%) oraz gatunki: acremonium strictum (8.5%), alternaria alternata (4.2%) i cladosporium cladosporioides (3.5%). rzadziej izolowano z liści gatunki patogeniczne botrytis cinerea (4.9%) oraz z rodzaju fusarium (3.2%) i ascochyta (2.1%). najmniej izolatów otrzymano z liści rutwicy w kombinacji z nawożeniem 80 kg p2o5 -1 × ha i 160 kg k2o × ha -1, jednak najczęściej izolowano tu patogeniczne gatunki z rodzaju ascochyta. nie wyizolowano tego grzyba z rutwicy uprawianej w kontroli, w przeciwieństwie do najliczniej występującego gatunku b. cinerea. 2014-01-01t11:48:06+0100 polish botanical society ecological characteristics of a hungarian summer truffle (tuber aestivum vittad.) producing area andrea csorbai gógán1, zsófia nagy2, zoltán dégi2, istván bagi3 and judit dimény1 1szent istván university, faculty of agricultural and environmental sciences, institute of horticultural technologies, páter károly str.1, h-2103, gödöllő, gogan.andrea@mkk.szie.hu 2nagykunság forest and timber management company, józsef attila str. 34, h-5000 szolnok 3truffleminers co. h-2335, kinizsi str. 1, taksony gógán a.cs., nagy z., dégi z., bagi i., dimény j.: ecological characteristics of a hungarian summer truffle (tuber aestivum vittad.) producing area. acta mycol. 47 (2): 133–138, 2012. hungary has outstanding environment for natural truffle production in some regions including plain and hilly areas. the most famous of all the natural summer truffle (tuber aestivum vittad.) habitats is the commonly called jászság region. this area is situated in the middle of hungary, between river danube and tisza. the flatland area is basically covered by river alluviums with main soils of chernozems, fluvisols, solonchaks and arenosols. climate of the region is typically continental: warm and dry summers and cold winters vary. the area is traditionally of agricultural use, although strong afforestation was made in the late 1950’s. the english oak (quercus robur l.) populations planted at that time gave a basis for current excellent truffle production. nowadays the region has proved to be the best natural summer truffle (t. aestivum) producing area of hungary with early season opening (june) and high quality truffles as early as august. in the research the best truffle producing forest blocks were selected for ecological investigation. results of the detailed site description showed uniform climate characteristics and dominance of english oak (q. robur) or mixed english oakturkey oak (quercus cerris l.) forests. soil types revealed differences from earlier findings: dominance of gleysols and water affected chernozems was declared. soil chemical parameters are in accordance with literature data: ph, organic matter and active carbonate content of the examined soils fall within the range indicated as the requirement of t. aestivum. key words: trufficulture, soil characteristics, climate, forest management acta mycologica vol. 47 (2): 133–138 2012 134 a. cs. gógán introduction consumption of truffles originates from ancient times in certain areas of the mediterranean region. from medieval centuries, use of black truffles gained ground in other european countries, based primarily on french and hungarian cookbooks of the period. truffles have been originated from natural forests from hundreds years and in some countries are still the main resources of truffle harvest. summer truffle (tuber aestivum), referred also burgundy truffle considered as the third most important one in europe and probably the most common truffle of marketable species (hall et al. 2007).it is a widespread truffle species in europe. due to its wide tolerance toward ecological factors, it is common from sweden to spain, from the united kingdom to russia (ceruti et al. 2003; granetti et al. 2005; tulasne 1851; wedén, danell 2001). in central europe hungary plays an important role in the truffle market with considerable yield of t. aestivum (gógán et al. 2007a). due to its wide tolerance toward environmental factors, the species is widespread in the country (bratek 2005). soil parameters can limit its distribution to soils rich in organic matter with variable lime content and neutral or lightly basic ph (gógán et al. 2007b; bratek 2005, 2008). materials and methods study area description. according to forest management it is called as tápió-zagyva territory, referred to the main rivers of the region, however, hungarian cadastral map refers the region jászság area. in our study, due to practical reasons, we fitted region borders to the management territory of nagykunság forest and timber management company, meaning a slight enlargement. the lowland, flat area is covered by river alluvium, relief is characterised by river deposits. the area is often threatened by inland inundations; floodplains are regularly covered by river floods. geological patterns is determined by middle miocene volcanic ash but rivers arriving from the north loaded fine sediments (mainly loam) to the surface in the pleistocene resulted in moorlands and marshland in the area. later 1-4 m deep loess and holocene alluvial warp arrived to the area. in some areas sandy soils appear. the most common soil types are calcareous chernozem and fluvisols but solonchaks and arenosols also occur. climate of the area is warm and dry with 2000 hours of annual sunshine and 10.2 c° yearly average temperature. average of the annual temperature minimum is –17 c°, maximum reaches 34 c°. average annual rainfall is measured 510-520 mm, of which 310 mm falls during vegetation period. the area is traditionally of agricultural use, about 75% of the surface covered by croplands. forest surface is under 5% (bulla 1962; danszky, rott 1964; dövényi 2010; marosi, somogyi 1990). before 1950’ forests were uncommon to the region. at that time, intensive reforestation started with english oak (q. robur) (25%), black locust (robinia pseudo-acacia l.) (28%) and populus species ecological characteristics 135 (31%) (danszky, rott 1964). the forest established in this period provides the base of current natural truffle habitats. site description method. in the study ecological characteristics of 20 truffle-producing natural habitats and data of a man-made truffle orchard were evaluated. hungarian forest site description methodology was applied to determine the characteristics of natural habitats and the truffle orchard. detailed site description included climate, micro-relief (elevation and orientation), hydrology and in-site soil description (soil type, type of mother rock, depth of organic layer and depth of soil layers based on 1,5 m deep soil-sampling holes). besides these parameters soil colour (wet munsell colour), granulometry, organic matter content (estimation based on soil colour), structure, compacted areas, quantity of roots, secondary soil formations, aggregations, estimate lime content and alkalinity for each soil layer were made. laboratory chemical and physical analysis parameters were defined considering forest site description methodology (áesz 2001, áesz 2010, babos et al. 1966; buzás 1988; buzás 1993; szodfridt 1993) and summer truffle (t. aestivum) ecological requirements (bratek 2005; chevalier, frochot 1997; granetti et al. 2005; morcillo et al. 2007; wedén et al. 2004). soil samples were collected from each soil layer, determined during site description. examined parameters were: ph (both h2o and kcl), lime content, organic matter content, soil conductivity (salt content), soil granulometry based on fao guidelines (1996). when necessary, further analysis was carried out in the case of saline and alkaline soils. in the article results of the laboratory analysis of the upper layer (a, a1, cca. 0-50 cm depth) were evaluated. results site-description results. all the examined natural truffle habitats is covered by english oak (q. robur) or mixed english oak-turkey oak (q. cerris) forests. field maple (acer campestre l.), black locust (r. pseudo-acacia l.), field elm (ulmus minor mill.), fraxinus spp., russian olive (elaeagnus angustifolia l.), common hackberry (celtis occidentalis l.), populus spp. and boxelder (acer negundo l.) are also present in the forests. elevation above sea level never exceeds 150 m, micro-relief is flat in all the examined cases. climate refers to forest steppe climatic category where annual rainfall less than 550-600 mm and annual average temperature is around 10.5 c°. 68% of the sampled areas are affected by temporary presence of groundwater surplus, however, most common water management types of the soils were considered to be medium dry (72%) or very dry (18%). water had heavily affected soil formation: gleysols (63%) and water affected chernozem (23%) are the most common soil types, in some cases solonchaks and solonetz soils occur (4.5 and 9%, respectively). mother rock is loess (50%), sandy loess (23%) or river sediment (23%) in most of the cases. results of the laboratory analysis. detailed granulometry of the examined soil samples resulted in the following textures: loam (40%), clay loam (50%) and clay (9%). only in one sampled area sand was present to define sandy loam soil. main parameters of the chemical analysis are listed in table 1. 136 a. cs. gógán discussion the climate of the examined area well fits in the requirement range of the species; however, for example, annual rainfall of jászság is resulted to be more close to the annual precipitation of drier gotland, sweden (514 mm) (wedén et al. 2004) and copenhagen, denmark (525 mm) (hall et al. 2007) than those of traditional french truffle producing regions (727 mm on average) (hall et al. 2007) or those of newly re-discovered german habitats (825 mm) (stobbe et al. 2012). similar to our research area, literature cites t. aestivum habitats of low elevation in most of the cases (sweden, denmark, uk, switzerland, france, croatia, and new-zealand), only some italian areas and spanish results occur in hilly regions (hall et al. 2007). although hydrology and water management play an important role in t. aestivum production, data in the topic is scarce. our study revealed that t. aestivum can occur on sites considered to be medium dry or dry, however, temporal water cover and soil texture (loam, clay loam and clay) can compensate together the problem of dryer summer periods. soil texture can play an important role of counterbalancing dry and hot summers and this could be the explanation for more heavy soils in the research area compared to loam-sandy loam soils of spain (morcillo et al. 2007) or sweden (wedén et al. 2004). french samples (chevalier, frochot 1997) show more variance considering soil texture while italian results (granetti et al. 2005) proved to be the closest to the soil of the study area. soil types are rarely determined in the table 1 chemical parameters of the examined soil samples (truff1-3: truffle orchard, forest1-20: natural truffle habitats) sample name ph (h2o) ph (kcl) salt content % active carbonate % organic matter % truff1 7.2 6.7 0.04 0.0 2.6 truff2 7.3 6.8 0.04 0.0 3.3 truff3 7.1 6.6 0.04 0.0 3.7 forest1 7.9 7.3 0.06 0.0 3.1 forest2 7.3 6.7 0.08 0.0 3.2 forest3 7.9 7.3 0.02 10.0 3.1 forest4 7.9 7.3 0.02 8.0 5.0 forest5 9.4 8.5 0.14 8.0 0.7 forest6 7.7 7.2 0.04 9.0 3.8 forest7 7.5 7.3 0.06 6.0 4.3 forest8 8.3 7.2 0.21 3.0 3.6 forest9 7.9 7.5 0.02 11.0 4.7 forest10 7.9 7.4 0.03 19.0 4.3 forest11 7.9 7.3 0.07 7.0 3.9 forest12 7.7 7.1 0.03 1.0 3.3 forest13 7.9 7.4 0.03 8.0 3.6 forest14 7.9 7.4 0.01 2.0 2.2 forest15 7.8 7.3 0.01 1.0 3.3 forest16 6.1 5.4 0.01 0.0 4.3 forest17 7.7 7.0 0.02 1.0 5.6 forest18 6.4 5.5 0.01 0.0 2.8 forest19 5.9 5.2 0.01 0.0 2.8 forest20 7.1 6.3 0.00 0.0 3.8 ecological characteristics 137 literature in connection with t. aestivum. chevalier, frochot (1997) solely mention rendzina and luvisol types as suitable for the truffle compared to our findings where mainly gleysols and water affected chernozems occur, however, appearance of solonchaks and solonetz and also the presence water soluble salt in some quantities in two samples (0.21% and 0.14%) indicates a certain tolerance toward lightly saline soils, previously not reported in literature. main chemical parameters of the soil of examined habitats agree on earlier findings (tab. 2). considering ph, except the abovementioned two saline soils, results fall within the previously indicated range. presence of active carbonate in the studied soils resulted very variable, ranging from 0 to 19% in the upper layer; carbonate-free soil samples of upper soil layers were present in cca. 40% of the examined cases, however, deeper soil layers (a1, a2, ac, etc.) counterbalanced lime-free upper soil layer in all the cases (data not shown). in water-affected chernozems, slight dissolution of active carbonate resulted in lower carbonate content of the upper layer of the soil but limited decalcification due to low annual precipitation and the soil-mixing effect of rich edaphic fauna can be traced. in the case of gleysols, modest decalcification also results in the presence of carbonates in the upper layers of the soil. due to the intensive evaporation of soil humidity in solonchaks and solonetz soils, water is moving upward and carries water-soluble salts, including less soluble carbonates which easily precipitate close to the surface. limerich mother rock (loess, sandy loess or river sediment) also plays an important role in the high carbonate content of the examined soils (stefanovits et al. 1999). we found generally lower organic matter content than that of literature mentions, especially compared to italian habitats, however, differences in organic matter due to distinct measurement methodology cannot be excluded. conclusion our findings generally supported the conception of the ecological demand of t. aestivum and some of the results even widened the tolerance of the species towards certain environmental conditions. acknowledgement. this study is included in the támop-4.2.1.b-11/2/kmr-2011-0003 project. table 2 main parameters of the examined soil samples compared to soil requirements of summer truffle (t. aestivum) (bratek 2005; chevalier, frochot 1997; granetti et al. 2005; morcillo et al. 2007; wedén et al. 2004) literature data compared to examined sites (min-max) ph (h2o) active carbonate (%) organic matter (%) italian sites (granetti et al. 2005) 7.0-7.87 0.9-20 11-14 spanish sites (morcillo et al. 2007) 7.16-8.45 0-71.43 2.98-23.52 swedish sites (wedén et al. 2004) 6.8-7.9 0.1-10.5 6.0-21.2 french sites (chevalier, frochot 1997) 7.1-8.0 0.4-52.0 4.4-21.1 hungarian sites (bratek 2005) 6.1-7.4 1-39 3.1-9.1 results of the study area 5.9-9.4 0-19 0.7-5.6 138 a. cs. gógán references áesz (állami erdészeti szolgálat) 2001. erdőtervezési útmutató, állami erdészeti szolgálat budapest. áesz (állami erdészeti szolgálat) 2010. erdőrendezési útmutató, termőhely felvétel kódjegyzéke és mellékletei. kivonat. állami erdészeti szolgálat, budapest. babos i., horváthné proszt s., járó z., király l., szodfridt i., tóth b. 1966. erdészeti termőhelyfeltárás és térképezés. akadémiai kiadó, budapest. bratek z. 2008. mycorrhizal research applied to experiences in plantations of mycorrhizal mushrooms, especially in central europe. proceedings of the sixth international conference on mushroom biology and mushroom products, bonn: 272–286. bratek z. 2005. a tuber aestivum kárpát-medencei termőhelyei (in:) g. chevalier, h. frochot, z. bratek (eds). az európai fekete szarvasgomba (burgundi szarvasgomba – tuber uncinatum chatin). első magyar szarvasgombász egyesület, budapest. bulla b. 1962. magyarország természeti földrajza. tankönyvkiadó, budapest. buzás i. (ed.) 1988. talajés agrokémiai vizsgálati módszerkönyv 2. a talajok fizikai-kémiai és kémiai vizsgálati módszerei. mezőgazdasági kiadó, budapest. buzás i. (ed.) 1993. talajés agrokémiai vizsgálati módszerkönyv 1. a talaj fizikai, vízgazdálkodási és ásványtani vizsgálata. inda 4231 kiadó, budapest. ceruti a., fontana a., nosenzo c. 2003. le specie del genere tuber. una revisione storica. museo regionale di scienze naturali, torino. chevalier g., frochot h. 1997. la truffe de bourgogne (tuber uncinatum chatin). éditions pétrarque, levallois-perret cedex. danszky i., rott, f. (eds). 1964. magyarország erdőgazdasági tájainak erdőfelújítási, erdőtelepítési irányelvei és eljárásai. általános irányelvek. erdőés termőhelytípus térképezés, budapest. dövényi z. (ed.) 2010. magyarország kistájainak katasztere. mta földrajztudományi kutató intézet, budapest. food and agriculture organization of the united nations 2006. guidelines for soil description, fourth edition, rome. gógán a. cs., bratek z., dimény j. 2007a. las trufas en hungría. (in:) s.r. domenech (ed.). truficultura: fundamentos y técnicas, ediciones mundi-prensa, madrid. gógán a. cs., bratek z., dimény j. 2007b. a new tool for rural development: truffle cultivation, cereal research communications 35 (2): 413–417. granetti b., de angelis a., materozzi g. 2005. umbria terra di tartufi, gruppo micologico ternano, terni. hall i.r., brown g. t., zambonelli a. 2007. taming the truffle. timber press, portland. marosi s., somogyi s. (eds) 1990. magyarország kistájainak katasztere. mta földrajztudományi kutató intézet, budapest. morcillo m., moreno b., pulido e., sánchez m. 2007. manual de truficultura andaluza. ed. gypaetus y consejería de medio ambiente. junta de andalucía. stefanovits p., filep gy., füleky gy. 1999. talajtan. mezőgazda kiadó, budapest. stobbe u, büntgen u., sproll, l., tegel w., egli s. 2012. spatial distribution and ecological variation of re-discovered german truffle habitats. fungal ecology 5 (5): 591–599. szodfridt i. 1993. erdészeti termőhelyismeret-tan. mezőgazda kiadó, budapest. tulasne ch. 1851. fungi hypogaei. apud friedrich klincksieck, paris. weden c., chevalier g., danell e. 2004. tuber aestivum (syn. t. uncinatum) biotopes and their history on gotland. sweden mycological research 108: 304–310. wedén ch., danell e. 2001. tuber aestivum uncinatum in sweden. actes du ve congrès international, science et culture de la truffe et des autres champignons hypoges comestibles. 4 au 6 mars 1999, aix-en-provence, france, federation française des trufficulteurs p. 4.247. 2014-01-02t12:09:33+0100 polish botanical society species of anamorphic fungi rare and new for poland iwona adamska department of plant pathology, university of agriculture słowackiego 17, pl-71-434 szczecin, iwonaadamska@interia.pl a d a m s k a i.: species of anamorphic fungi rare and new for poland. acta mycol. 42 (1):79-84, 2007. morinia pestalozzioides, seimatosporium hypericinum, septoria artemisiae, s. artemisiaemaritimae, s. achilleicola and s. symphyti, fungi not recorded in poland before, are described and illustrated. the species were found during studies on the occurrence of parasitic fungi conducted in selected sites in the słowiński national park and in the western pomerania between 2001 and 2004. key words: morinia, seimatosporium, septoria, słowiński national park, parasitic fungi introduction achillea ptarmica, artemisia vulgaris, hypericum perforatum, and symphytum officinale are plants of considerable pharmaceutical importance in the climatic zone of poland. their green parts (a. ptarmica, ar. vulgaris and h. perforatum), roots (a. ptarmica and s. officinale), and capitula (a. ptarmica) are used in drug and cosmetic production ( p o d b i e l k o w s k i , s u d n i k -w ó j c i k o w s k a 2003) . these plants may be encountered in shrubs and meadows, on hills, and riverbanks, and occur commonly in the lowland part of poland. during studies conducted in the słowiński national park (snp) and the western pomerania (wp), achillea ptarmica, artemisia vulgaris, ar. campestris subsp. sericea, hypericum perforatum, and symphytum officinale were frequently recorded to be infected by parasitic fungi. among the fungi identified, morinia pestalozzioides, septoria achilleicola, s. artemisiae, s. artemisiae-maritimae and s. symphyti occurred, fungi previously not reported from poland. the species are described and illustrated below. acta mycologica vol. 42 (1): 79-84 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 80 i. adamska material and methods in the years 2001 to 2004, diseased stems and leaves of plants growing in the diantho-armerietum elongatae krausch 1959, helichryso-jasionetum litoralis libb. 1940, myrico-salicetum auritae (allg. 1922) r. tx. et pass. 1961, and fraxino-alnetum w. mat. (1952) 1987 plant associations of the snp and wp were collected. in the laboratory, they were first air dried and then placed into paper envelopes. to identify the parasitic fungi associated with the plant parts collected, thin cuttings were taken from a transverse section of the fungal structures revealed and the plant tissue infected using a safety razor. subsequently, the cuttings were mounted in a drop of lactic acid placed on a microscopic slide, covered with a cover slip, and observed under the axiolab zeiss compound microscope. the structures of the fungi found were measured with a micrometric eyepiece and a screwmicrometer. the nomenclature of plant associations is according to m a t u s z k i e w i c z ( 2001) , and that of plant species follows m i r e k et al. ( 2002). the plant species sampled were identified according to s z a f e r , k u l c z y ń s k i , p a w ł o w s k i (1969). the fungi found were determined after b r a n d e n b u r g e r (1985), e l l i s and e l l i s ( 1987) , s u t t o n ( 1980). all the specimens of plants and fungi discussed below are deposited at the department of plant pathology university of agriculture in szczecin. results and discussion six anamorphic fungal species new and rare for poland were found. they colonised leaves of hypericum perforatum, artemisia campestris subsp. sericea, a. vulgaris, achillea ptarmica, and symphytum officinale. symbols: caen – carici arenariae-empetretum nigri dae – diantho-armerietum elongatae hejl – helichryso-jasionetum litoralis msa – myrico-salicetum auritae fral – fraxino-alnetum morinia pestalozzioides berl. & berk. spots on dead stems, subcircular, grey to brown, 2-4 mm diam. acervuli circular, dark brown to black, 150 μm diam. conidia muriform, with 5 transverse septa and 2-3 vertical septa, 18-25 x 9-12 μm. medial cells brown, terminal cells lightolive to hyaline. apical cell with 2-3 unbranched, hyaline setulae, 10-12 μm long. on artemisia campestris l. subsp. sericea: spn, łeba, caen vi 2002 (figs 1 a-c). the conidial dimensions in the material found in the snp slightly diverged from those given by s a c c a r d o ( 1892; conidia 22-24 x 8-10 μm; apical setulae 14-20 x 1 μm) and s u t t o n (1980; conidia 16-22 x 5-7 ųm; apical setulae 12-19 ųm long). distribution. morinia pestalozzioides has so far been reported from italy (s u t t o n 1980). remarks. according to s u t t o n (1980), a. camphorata vill. is another host plant of m. pestalozzioides. species of anamorphic fungi 81 seimatosporium hypericinum (ces.) b. sutton spots randomly distributed on the upper leaf side, irregular, grey to pale brown, 3-6 mm diam. acervuli brown to black, 200-250 μm diam. conidia curved, falcate, 4-celled conidia, 14-15 x 6.5-7.5 μm. two medial cells 4-5 μm long, olive to lightolive. terminal cells 1-2 μm long, hyaline, with 2 unbranched, straight or curved, hyaline setulae, 12-14 μm long. on hypericum perforatum l.: wp, karkowo near kołobrzeg, dae vi 2002 (figs 1 d-f). the acervular diameter of the specimen found by the present author was convergent with that given by s u t t o n ( 1980; diameter above 200 μm). the conidial dimensions in the material found in the snp slightly diverged from those given by e l l i s and e l l i s (1987; 15-19 x 4.5-5.5 μm) and s u t t o n (1980; 15-18 x 4.5-5.5 μm). the colour of medial cells in the conidia found in the material collected in the snp slightly diverged from that given by e l l i s and e l l i s (1987) and s u t t o n (1980). according to these authors, these cells were light-brown, while they were lighter, mostly olive, in the material collected in the snp. distribution. poland. seimatosporium hypericinum has already been reported to occur in the lublin region (z i m o w s k a 2002, 2004; z i m o w s k a , m a c h o w i c z s t e f a n i a k 1999, 2004) and in the western pomerania (a d a m s k a 2006). other regions. seimatosporium hypericinum has so far been reported from italy, sweden, germany, great britain, france (s u t t o n 1980), and australia (fa r r et al. 2007). remarks. apart from h. perforatum, another host plant of s. hypericinum is h. tetrapterum fries (e l l i s , e l l i s 1987). septoria achilleicola melnik spots on the upper leaf side, ovoid to circular, pale brown to dark brown, 3-5 mm diam. pycnidia globose, brown to dark brown, 125 μm diam., with a circular ostiolum (15-20 μm), immersed in the leaf tissue. conidia hyaline, filamentous, 4-7 celled, 3237.5 x 2-2.5 (-4) μm. on achillea ptarmica l.: spn, kluki, msa viii 2001. the conidial dimensions were within the range given by te t e r e v n i k o v a b a b a j a n (1987; 14-40 x 1-2 μm); however, the conidia of the specimen collected in the snp were slightly shorter than those described by b r a n d e n b u r g e r (1985; 36-60 x 1.5-1.8 μm). the number of septa in the conidia was greater than that given by te t e r e v n i k o v a b a b a j a n (1987; 1-2-septate), but close to that given by b r a n d e n b u r g e r [1985; 2-4-(6)-septate]. distribution. septoria achilleicola has been reported from the european part of russia, kazakhstan, and asia (te t e r e v n i k o v a b a b a j a n 1987). remarks. according to te t e r e v n i k o v a b a b a j a n (1987), another host plant of s. achilleicola is a. salicifolia besser. other fungal species of the genus septoria infecting plants of the genus alchemilla are s. moschatae f. mangenot (b r a n d e n b u r g e r 1985), s. millefolii (oudem.) grove, and s. ptarmicae pass. (te t e r e v n i k o v a b a b a j a n 1987). septoria artemisiae pass. spots distributed on the upper leaf side, subcircular, yellow to pale brown. pycnidia numerous, globose, brown, 120-150 μm diam., with a circular ostiolum (35 μm diam.), immersed in the leaf tissue. conidia hyaline, filamentous, aseptate, 3582 i. adamska 40 x 1-2 μm. on artemisia campestris l. subsp. sericea: spn, łeba, hejl viii 2002, a. vulgaris l.: spn, gać, fral ix 2001. the conidial dimensions of the specimens found by the author of this paper were similar to those given by b r a n d e n b u r g e r (1985; 30-33 (57) x 1-1.5 μm), c e j p and j e c h o v a (1967; 19.5-57 x 1-1.5 μm), and te t e r e v n i k o v a b a b a j a n (1987; 20-45 x 1-1.5 μm). distribution. septoria artemisiae has been observed in the european part of the former soviet union, the netherlands, germany, romania, the czech republic, slovakia, hungary, italy, the us, asia (te t e r e v n i k o v a b a b a j a n 1987), as well as in bulgaria, korea, china and india (fa r r et al. 2007) . remarks. apart from a. vulgaris, other plant hosts of s. artemisiae are a. austriaca jacq., a. dracunculus l., and a. scoparia w. et k. (te t e r e v n i k o v a b a b a j a n 1987). according to b r a n d e n b u r g e r (1985), plants of the genus artemisia are infected by three species of the genus septoria (i. e., s. artemisiae pass., s. artemisiae-maritimae lobik, and s. globosa strasser), and not by five (s. artemisiae, s. artemisiaemaritimae, s. artemisiana garb., s. moeszii smarods, s. tabacina died. var. tabacina, and s. tabacina died. var. dracunculina d. bab.) as te t e r e v n i k o v a b a b a j a n (1987) found. b r a n d e n b u r g e r (1985) considered s. moeszii and s. artemisiana to be synonyms of s. globosa. according to te t e r e v n i k o v a b a b a j a n (1987), a. vulgaris is sometimes also affected by s. artemisiana and s. tabacina var. tabacina. septoria artemisiae-maritimae pass. spots on the upper leaf side, circular, grey to pale brown, 1-2 mm diam. pycnidia flattened, pale brown to brown, 65-95 μm diam., with a circular ostiolum (20-28 μm diam.), immersed in the leaf tissue. conidia narrowly cylindrical, hyaline, aseptate or 3-5-celled, 25-30 (-35) x 2.5-4 μm. on artemisia campestris l. subsp. sericea: spn, łeba, hejl ix 2002 (figs 1 g-i). the conidial dimensions of the specimens found by the author of this paper were similar to those given by b r a n d e n b u r g e r (1985; 20-26 x 2.2-3.6 μm) and te t e r e v n i k o v a b a b a j a n (1987; 19.8-26.3 x 3-4 μm). distribution. septoria artemisiae-maritimae has already been observed in the european part of the former soviet union and asia (te t e r e v n i k o v a b a b a j a n 1987). remarks. according to te t e r e v n i k o v a b a b a j a n (1987), another host plant of s. artemisiae-maritimae is a. maritima l. var. salicina. septoria symphyti cejp spots on the upper leaf side, ovoid to irregular, brown to dark brown, 5-15 mm diam. pycnidia globose, brown, 80-135 μm diam., immersed in the leaf tissue. conidia filamentous, hyaline, 4-septate, 37.5 x 1-1.5 μm. on symphytum officinale l.: spn, gać, fral ix 2004. the pycnidium diameter and the conidial length in the material collected in the snp were similar to those of the specimens described by b r a n d e n b u r g e r (1985; 90-110 μm diam and 24-48 μm long, respectively). the conidial length and the number of septa in the conidia (3-4-septate) also were similar to those given by b r a n d e n b u r g e r (1985); however, the conidia in the material collected in the species of anamorphic fungi 83 snp were narrower (37.5 x 1-1.5 μm) than those characterized by b r a n d e n b u r g e r (1985; 24-48 x 1.7-2.6 μm). distribution. septoria symphyti has been observed in the czech republic, slovakia, and bulgaria (fa r r et al. 2007). remarks. according to b r a n d e n b u r g e r (1985), s. symphyti is the only species of the genus septoria occurring on plants of the genus symphytum in europe. acknowledgement. the article was completed as part of studies financed by the state committee for scientific research, grant no 3 po4 017 23. references a d a m s k a i. 2006. choroby grzybowe ziół i zwalczanie ich sprawców. progress in plant protection 46 (2): 654–656. b r a n d e r b u r g e r w. 1985. parasitische pilze an gefäβpflanzen in europa. fischer. stuttgart, new york. c e j p k . , j e c h o v a v. 1 9 6 7 . beitrag zur kenntnis einiger tschechoslowakischen arten der gattung septoria fries. acta musei nationalis pragae. 23 (4): 101–123. e l l i s m . b . , e l l i s j . p. 1987. microfungi on land plants. an identification handbook. croom helm. f a r r d . f. , r o s s m a n a . y . , p a l m m . e . , m c c r a y e . b . 2007. fungal databases, systematic botany & mycology laboratory, ars, usda. retrieved march 14, 2007 from http://nt.ars-grin.gov/ fungaldatabases/. m a t u s z k i e w i c z w. 2001. przewodnik do oznaczania zbiorowisk roślinnych polski. pwn, warszawa. m i r e k z . , p i ę k o ś m i r k o w a h . , z a j ą c a . , z a j ą c m. 2002. flowering plants and pteridophytes of poland. a checklist. biodiversity of poland 1: 1–442. p o d b i e l k o w s k i z . , s u d n i k w ó j c i k o w s k a b. 2003. słownik roślin użytkowych. pwril. warszawa. s a c c a r d o p. a . 1892. sylloge fungorum 10: 508. s u t t o n b . c . 1980. the coelomycetes. fungi imperfecti with pycnidia, acervuli and stromata. commonwealth mycological institute. kew, surrey, england. s z a f e r w. , k u l c z y ń s k i s . , p a w ł o w s k i b . 1969. rośliny polskie. pwn warszawa. te t e r e v n i k o w a b a b a j a n d . n . 1987. griby roda septoria w sssr. wyd. akad. nauk armeńskiej ssr. erevan. z i m o w s k a b. 2002. wpływ warunków hodowli na wzrost, zarodnikowanie i tworzenie struktur morfologicznych przez seimatosporium hypericinum (ces.) sutton. acta agrob. 55: 401–410. z i m o w s k a b . 2004. occurrence, biology and some morphology elements of seimatosporium hypericinum, a pathogen of st. johhn’s wort (hypericum perforatum). phytopathol. pol. 34: 41–50. z i m o w s k a b . , m a c h o w i c z s t e f a n i a k z . 1999. grzyby występujące na niektórych roślinach zielarskich uprawianych w woj. lubelskim. bioróżnorodność w fitopatologii europejskiej na przełomie wieków. streszczenia. poznań: 155. z i m o w s k a b ., m a c h o w i c z s t e f a n i a k z . 2004. grzyby zagrażające uprawie dziurawca zwy-zwyczajnego (hypericum perforatum l.) w województwie lubelskim. acta sci. pol., hortorum cultus 3: 61–74. 84 i. adamska rzadkie i nowe dla polski gatunki grzybów anamorficznych s t r e s z c z e n i e w latach 2001-2004 w wybranych stanowiskach słowińskiego parku narodowego i pomorza zachodniego prowadzono badania nad występowaniem grzybów pasożytniczych. w trakcie badań znaleziono morinia pestalozzioides, seimatosporium hypericinum, septoria artemisiae, s. artemisiae-maritimae, s. achilleicola i s. symphyti, gatunki nowe dla polski. w artykule opisano i zilustrowano znalezione gatunki grzybów. 2014-01-01t11:45:21+0100 polish botanical society new phoma species on leonurus cardiaca beata zimowska department of plant pathology, agricultural university leszczyńskiego 7, pl-20-069 lublin beata.zimowska@ar.lublin.pl z i m o w s k a b.: new phoma species on leonurus cardiaca. acta mycol. 42 (1):119-123, 2007. two species of phoma obtained from motherwort leonurus cardiaca l., during mycological analyses attendant upon three-years study connected healthiness of the plants. isolates of phoma capitulum were obtained from roots, whereas phoma septicidalis from roots and leaves. description in vitro, photos of morphological structures and distribution of phoma spp. are given. this is the first report of p. capitulum and p. septicidalis on motherwort in poland. key words: phoma, occurrence, morphology, leonurus cardiaca introduction phoma spp. (sphaeropsidales) constitute a group of fungi with very differentiated biological specialization (m a r c i n k o w s k a 1995). they include saprophytic species, developing on different substrata, secondary pathogens of plants with polyphagous character of parasitism and specific pathogens of cultivated plants. the group of plants from which fungi from genus phoma were isolated includes for example herbaceous plants (m a c h o w i c z -s t e f a n i a k et al. 2002; m a c h o w i c z -s t e f a n i a k et al. 2004). phoma dictamnicola boerema, de gruyter et noordel., p. glaucii brun., p. multirostrata var. microspora (allescher) boerema from the leaves, stems and roots of common thyme, while p. eupatorii died. was obtained from creemocarps (m a c h o w i c z -s t e f a n i a k , z i m o w s k a 2000; m a c h o w i c z -s t e f a n i a k et al. 2002). p. exigua var. exigua desm. and p. glomerata (corda) wollenw. et hochapf. were isolated from the roots, stems and leaves of common thyme, common balm and st. john’s wort (z i m o w s k a , m a c h o w i c z -s t e f a n i a k 2004; m a c h o w i c z -s t e f a n i a k et al. 2004). isolates of p. strasseri moesz. were obtained from the stems and rhizomes of peppermint showing the signs of black rot (z i m o w s k a , m a c h o w i c z -s t e f a n i a k 2005). in recent years the species of phoma capitulum pawer, mathur et thirumalachar (p. ostiolata var. ostiolata pawer, mathur et thirumalachar, p. ostiolata var. brunnea pawer, mathur et thirumalachar) as well as phoma septicidalis boerema acta mycologica vol. 42 (1): 119-123 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 120 b. zimowska (pyrenoacheta telephii alesch., phoma telephii (vestergr.) kestern), which have not been described earlier, on this plant, have been isolated from leonurus cardiaca l. (d e g r u y t e r , n o o r d e l o o s 1992; d e g r u y t e r , b o e r e m a 2002). in accessible literature these species are enumerated as commonly occurring saprotrophs developing on different substrata, and p. septicidalis can also cause necrosis of the leaves of plants from the family gramineae (d e g r u y t e r , n o o r d e l o o s 1992; d e g r u y e r , b o e r e m a 2002). considering the key importance of phoma spp. morphology, with the proper differentiation of taxa, the present paper deals with the identification of the enumerated two species of fungi made on the basis of thorough studies of macroand microscopic properties of these organisms in cultures in vitro. material and methods the study material consisted of isolates lc 186, lc 259, lc 368, lc 544 and lc 56 p. capitulum obtained from the roots and isolates lc 238, lc 79, lc 139 and lc 225 p. septicidalis from the roots and leaves of 2-year-old motherwort. the fungi cultures used were obtained as a result of the mycological analysis (z i m o w s k a , m a c h o w i c z -s t e f a n i a k 2004) accompanying the three-year-long studies (2004-2006) on plants’ healthiness that were conducted on production plantation in the communes of fajsławice and dziecinin in the lublin district (z i m o w s k a unpublished). identification of the obtained isolates and the taxa they belong to was carried out on the basis of a thorough analysis of morphological structures, macroscopic properties of colonies and biochemical features conducted in vitro on standard media (maltose ma, oat agar oa and cherry agar ca) after 7 and 14 days of incubating the cultures in a thermostat at the temperature of 22oc (d e g r u y t e r , n o o r d e l o o s 1992; z i m o w s k a , m a c h o w i c z -s t e f a n i a k 2005). the studied fungi cultures were marked on the basis of a monograph by d e g r u y t e r and n o o r d e l o o s (1992) and d e g r u y t e r and b o e r e m a (2002). results the studies conducted on the growth of the analyzed isolates of p. capitulum on standard media showed that the cultures of the fungus on ma after 7 days reached the diameter of 38-40 mm, while after 14 days 75 mm. the hyphae of the air mycelium formed a flocky structure of white or light olive-grey colour. the reverse was peach-salmon colour. the edge of cultures of all the studied isolates was characterized by regular growth. the growth of p. capitulum isolated on oa was slightly poorer than on ma since the diameter of the colony measured after 7 days was 36 mm, while after 14 days it ranged from 65 to 70 mm. the colonies were white-creamy with a salmon reverse, while the hyphae of the air mycelium formed a velvet-flocky structure. the smallest diameter of colonies was observed on medium ca. after 7 days it ranged from 27 to 29 mm, while after 14 days – from 53 to 58 mm. in the case of most isolates, on this medium the edge of a colony was characterized by irregular growth. the hyphae of the air mycelium were white and they formed a flocky structure. the reverse of the colony was of light salmon colour. phoma species on leonurus cardiaca 121 numerous pycnidia on oa centered in concentric circles or sectors. they were round, olive to olive-black, mostly smooth or covered with scarce hyphal outgrowths (fig. 1). from 1 to 3 ostioles, from which grey conidial exudates, were observed on the surface of the pycnidia. the latter were mostly formed on the surface of agar or they were partly immersed in it. their size was from 50 to 100 μm, on average (tab. 1). the conidia were hyaline, round or slightly ellipsoidal, and they contained 1 to 2 guttules, their size ranging from 3.7 to 2.5 μm (fig. 2, tab. 1). in the case of all the studied isolates of p. capitulum, the reaction with 1n naoh was negative. besides, none of the isolates formed chlamydospores. after 7 days of growth p. septicidalis isolates on ma medium formed colonies with the diameter ranging from 20 to 38 mm, while after 14 days the diameter ranged from 41 to 70 mm. the hyphae of the air mycelium formed a compact felt-like structure of rosy-wine colour and the reverse of the same colour. a slightly bigger diameter was observed in the case of the growth of the colonies of p. septicidalis isolates on oa medium. after 7 days it ranged from 21 to 40 mm, while after 14 days – from 50 to 57 mm. the structure of the air mycelium was the same as on ma medium. the central part of the colony was olive-grey, while the edge had rosy-wine colour. on ca medium the fungus colonies reached the diameter from 20 to 29 mm after 7 days, while after 14 days it ranged from 40 to 60 mm. the structure of the ta b l e 1 characteryzation of pycnidia and conidia of phoma capitulum and phoma septicidalis (mean for 5 isolates) author p. capitulum p. septicidalis pycnidia conidia pycnidia conidia shape and dimension in μm arrangement and structure of walls surface shape and dimension in μm shape and dimension in μm arrangement and structure of walls surface shape and dimension in μm own data globose with 1 to 3 ostioles, 50 – 100 on the agar in concentric zones or sectors, with softly hyphal outgrowts globose to shortly ellipsoid, with 1 to 2 guttules, 3.7 x 2.5 globose with 1 to 2 ostioles 65-160 mainly on the agar but also in the agar, long setae spread over the surface ellipsoidal with several small guttules, 4.4 x 2.2 de gruyter and noordeloos 1992 globose with 1 to 3 ostioles, 60-105 mostly on the agar but also partly or entirely in the agar in concentric zones or sectors, with hyphal outgrowts broadly and shortly ellipsoid with 1 to 2 guttules, 3.8 x 2.6 de gruyter and boerema 2002 globose to subglobose with 1 to 2 ostioles, 70-170 mainly on the agar, with long setae spread mainly over the upper surface subglobose to ellipsoidal with several small or large guttules, 4.5 x 2.3 122 b. zimowska air mycelium, the colour of the obverse and the reverse was similar to the colonies growing on oa medium. the edge of the studied isolates of p. septicidalis on all the three media was characterized by irregular growth. detailed observations on oa medium showed that the pycnidia of the studied fungus isolates got formed individually or in small groups. they were most frequently round, honey-olive to black-olive one, and their walls were covered by very numerous setae with the length ranging from 180 to 210 μm (fig. 3, tab. 1). 1 to 2 ostioles, from which white drops of conidial exudates, were observed on the surface of the pycnidia. the size of the pycnidia ranged from 65 to 160 μm (tab. 1). the conidia were hyaline, 1-cell, ellipsoidal, with a few gutteles with the dimensions of 4.4 x 2.2 μm (fig. 4, tab. 1). in the case of all the studied isolates of p. septicidalis, the studies observed a positive reaction with 1n naoh on ma and oa media. the edge of the colonies was of violet-purple. besides, fragmentation of the mycelium cells characteristic of this species was visible on the hyphae of the mycelium, especially in older cultures (fig. 5). none of the studied isolates of the fungus formed chlamydospores. discussion thanks to the studies in vitro on the morphology of pycnidia and conidia and on the character of the growth of the analyzed cultures it was possible to distinguish two discussed species of phoma and classify them into the proper sections. results of the studies as compared to the descriptions contained in the monographs of fungi from the genus phoma made it possible to place p. capitulum isolates in part i of phoma section since they include studies on taxa with very small dimensions of conidia, with the length not exceeding 5.5 μm (d e g r u y t e r , n o o r d e l o o s 1992). although the dimensions of the conidia in fungi from genus phoma play a secondary diagnostic importance (d o r e n b o s c h 1970), in the case of the species belonging to phoma section this property acquires a special significance due to the division of this section into three parts performed on the basis of the conidia dimensions (d e g r u y t e r , n o o r d e l o o s 1992; d e g r u y t e r et al. 1993; d e g r u y t e r et al. 1998). on the basis of a detailed analysis of morphological structures, especially pycnidia, the studied isolates of p. septicidalis were classified into the section of paraphoma. a characteristic feature of the species described by the dutch mycologists is the formation of pycnidia with the walls covered with setae, which are formed on their whole surface or they occur only around the ostiole (d e g r u y t e r , b o e r e m a 2002). the conducted studies confirmed the correctness of the assumption presented in the 1930’s and 1940’s by woollenweber, hochapfel and dennis (d e n n i s 1946), and later continued by the mycologists from a dutch station of plant protection in wageningen (b o e r e m a , b o l l e n 1975) that defining the taxa within phoma spp. is possible only on the basis of constant morphological features that are observed in vitro in the cultures developing in standard conditions. phoma species on leonurus cardiaca 123 references b o e r e m a g. h., b o l l e n g. j. 1975. conidiogenesis and conidial septation as differentiating criteria between phoma and ascochyta. persoonia 8: 111–144. d e n n i s r. w. g. 1946. notes on some british fungi ascribed to phoma and related genera. trans. br. mycol. soc. 29: 11–42. d o r e n b o s c h m . m . j . 1970. key to nine ubiquitous soliborne phomalike fungi. persoonia 6: 1–14. g r u y t e r j . d e , n o o r d e l o o s m . e . 1992. contributions towards a monograph of phoma (coelomycetes) i. 1. section phoma: taxa with very small conidia in vitro. persoonia 15 (1): 71–92. g r u y t e r j . d e , n o o r d e l o o s m . e . , b o e r e m a g . h . 1993. contributions towards a monograph of phoma (coelomycetes) i. 2. section phoma: additional taxa with very small conidia and taxa with conidia up to 7 μm long. persoonia 15 (3): 369–400. g r u y t e r j . d e , n o o r d e l o o s m . e . , b o e r e m a g . h . 1998. contributions towards a monograph of phoma (coelomycetes)-i. 3. taxa with conidia longer than 7 μm. persoonia 16 (4): 471-490. g r u y t e r j . d e , b o e r e m a g . h . 2 0 0 2 . contributions towards a monograph of phoma (coelomycetes) viii. section paraphoma: taxa with setose pycnidia. persoonia 17 (4): 541–561. m a c h o w i c z s t e f a n i a k z . , z i m o w s k a b . 2000. grzyby przenoszone przez nasiona ro�lin zie-s t e f a n i a k z . , z i m o w s k a b . 2000. grzyby przenoszone przez nasiona ro�lin zie-s t e f a n i a k z . , z i m o w s k a b . 2000. grzyby przenoszone przez nasiona ro�lin ziez . , z i m o w s k a b . 2000. grzyby przenoszone przez nasiona ro�lin zie-z i m o w s k a b . 2000. grzyby przenoszone przez nasiona ro�lin zieb . 2000. grzyby przenoszone przez nasiona ro�lin zielarskich. acta agrobot. 53 (2): 25–38. m a c h o w i c z s t e f a n i a k z . , z i m o w s k a b . , z a l e w s k a e . 2002. grzyby zasiedlające różne organy tymianku wła�ciwego thymus vulgaris l. uprawianego na lubelszczyźnie. acta agrobot. 55 (1): 185–197. m a c h o w i c z s t e f a n i a k z . , z a l e w s k a e . , z i m o w s k a b . 2004. grzyby zasiedlające nadziemne organy melisy lekarskiej (melissa officinalis l.) i tymianku wła�ciwego (thymus vulgaris l.) uprawianych na lubelszczyźnie. folia univ. agric. stein., agriculture 239 (95): 229–232. m a r c i n k o w s k a j . 1995. nowoczesne spojrzenie na systematykę rodzaju phoma. wiad. bot. 39 (3/4): 47–52. z i m o w s k a b . , m a c h o w i c z s t e f a n i a k z . 2004. grzyby zagrażające uprawie dziurawca zwy-zwyczajnego (hypericum perforatum l.) w województwie lubelskim. acta sci. pol., hortorum cultus 3 (1): 61–74. z i m o w s k a b . , m a c h o w i c z s t e f a n i a k z . 2005. charakterystyka izolatów phoma strasseri nie notowanego w polsce patogenu mięty pieprzowej (mentha piperita l.). acta agrobot. 58 (2): 151–162. nowe gatunki z rodzaju phoma na serdeczniku pospolitym s t r e s z c z e n i e podczas analizy mikologicznej towarzyszącej trzyletnim badaniom (2004-2006), nad zdrowotno�cią ro�lin serdecznika pospolitego (leonurus cardiaca), uzyskano nie opisywane wcze�niej na tej ro�linie gatunki z rodzaju phoma. izolaty phoma capitulum wyosobniono z korzeni, za� phoma septicidalis z korzeni i li�ci serdecznika pospolitego. na podstawie szczegółowych obserwacji cech makroskopowych oraz mikroskopowych obserwowanych in vitro w kulturach na standardowych podłożach, izolaty p. capitulum zakwalifikowano do sekcji phoma a izolaty p. septicidalis do sekcji paraphoma. fig. 1. pycnidia of phoma capitulum (sem x 250). phot. m. wróbel. fig. 2. conidia of phoma capitulum (sem x 5100). phot. m. wróbel. fig. 3. setose pycnidium of phoma septicidalis (sem x 300). phot. m. wróbel. fig. 4. conidia of phoma septicidalis (sem x 2600). phot. m. wróbel. fig. 5. fragmentation of hyphae of phoma septicidalis (sem x 3000). phot. m. wróbel. 2014-01-01t11:45:39+0100 polish botanical society armillaria species in coniferous stands anna żółciak forest research institute, sękocin stary, braci leśnej 3, pl-05-090 raszyn, a.zolciak@ibles.waw.pl ż ó ł c i a k a.: armillaria species in coniferous stands. acta mycol. 42 (2):211-217, 2007. identification of the armillaria species in selected coniferous stands (scots pine stands, norway spruce stands and fir stands) was the aim of the work carried out on the basis of mating tests and consideration of macroscopic traits of fruit bodies. one species of armillaria [a. ostoyae (romagnesi) herink] was found in scots pine stands, three species [a. ostoyae, a. cepistipes velenovský and a. borealis marxmüller et korhonen] were found in norway spruce stands and two species [a. ostoyae and a. cepistipes] were found in fir stands. key words: coniferous stands, armillaria ostoyae, a. cepistipes, a. borealis introduction armillaria root infection is the most important disease affecting mainly young scots pines (pinus sylvestris l.) planted often in/after mixed stands in the north-east part of poland and norway spruce (picea abies karst.) stands in the south part of the country (m a ń k a 1953, 1998; tw a r o w s k i , tw a r o w s k a 1959; tw a r o w s k a 1965; r y k o w s k i 1985; c a p e c k i 1994, 1997; s i e r o t a 2001; l e c h 2003; m a ń k a et al. 2003). it is caused by armillaria species. from seven european armillaria species, five were identified in poland: a. borealis marxmüller et korhonen, a. cepistipes velenovský, a. ostoyae (romagn.) herink, a. mellea (vahl: fr.) kummer and a. gallica marxmüller et romagnesi (ż ó ł c i a k 1999). the objectives of the present study were to identify armillaria species in selected coniferous stands: scots pine stands, norway spruce stands and fir stands in poland and to determine the occurrence of armillaria species considering forest habitat (according to polish forest site typology) and stand age classes. materials and methods material (samples) consisted of fruit bodies of armillaria, fragments of wood colonized by mycelium of armillaria and rhizomorphs. samples were taken from experimental one-time-sampled plots of 500 m2 each (20x25 m) established in production stands: scots pine stands, norway spruce stands and fir stands. the stands were acta mycologica vol. 42 (2): 211-217 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 212 a. żółciak chosen on the basis of data obtained from questionnaires of the annual forest state assessments informing about the area of stands infested by root rot of armillaria, inventory documents and the author’s own observations. one hundred and five plots were established in the scots pine stands, situated within the territory of 52 forest districts (fig. 1a). on each plot 1 to 10 samples were taken in 1985, 1989-94 and 1996-1998. fifty plots were established in the norway spruce stands, situated within the territory of 17 forest districts, 1 in gorce national park and 1 in krynica experimental forest. on each plot 1 to 15 samples were taken in 1981, 1989, 1991, 1994, 1996-98 and 2001-03. fifteen plots were establish in the fir stands, situated within the territory of 5 forest district, 1 – in gorce national park and 1 – in rotocze national park. on each plot 1-12 samples were taken in 1991, 1994, 1996 and 1998. characterization of the stands and samples is presented in table 1. the identification of armillaria isolates was performed by using mating tests (k o r h o n e n 1978). fruit bodies of armillaria were identified on the basis of the macroand microscopic features (r o m a g n e s i , m a r x m ü l l e r 1983). ta b l e 1 characterization of stands, from which samples of armillaria were taken stands age of stands type of forest habitat* proportion of main species (%) number of the experimental plots samples obtained isolates (fruit bodies/ colonized wood/ rhizomorphs) identified using mating tests samples (fruit bodies) identified of the macroand microscopic features scots pine 1-160 fcf, fmcf, fmbf, mmcf, mmbf 100 (scots pine) 105 356 176 (71/105/-) 180 norway spruce 1-120 fmcf, fbf, fmbf, muf, mmf, mf 60-100 norway spruce 50 228 114 (86/27/1) 114 fir 41-140 fbf, uf, mmf, mf 50-100 (fir) 15 53 25 (23/2/-) 28 total 170 637 315 (180/134/1) 322 explanations: fresh coniferous forest (fcf), fresh mixed coniferous forest (fmcf), moist mixed coniferous forest (mmcf), fresh broadleaved forest (fbf), fresh mixed broadleaved forest (fmbf), upland forest (uf), mixed upland forest (muf), moist mixed broadleaved forest (mmbf), mountain forest (mf), mixed mountain forest (mmf). armillaria species 213 fig. 1. a) distribution of experimental plots. b) distribution of armillaria species in scots pine stands, norway spruce stands and fir stands. 214 a. żółciak results a total of 637 samples were collected in the investigated coniferous stands (356 samples in scots pine stands, 228 samples in norway spruce stands and 53 samples – in fir stands). the proportions of armillaria samples found in the coniferous stands were calculated (tabs 2 and 3). ta b l e 2 percentage of samples of armillaria from coniferous stands: scots pine, norway spruce and fir considering different types of forest habitat stands type of forest habitat fcf (%) fmcf (%) mmcf (%) fbf (%) fmbf (%) mmbf (%) uf (%) muf (%) mmf (%) mf (%) scots pine 9,3 78,4 5,6 0,8 4,8 1,1 norway spruce 0,9 2,6 12,7 13,6 36,4 33,8 fir 3,8 45,3 3,8 47,1 ta b l e 3 percentage of samples of armillaria from coniferous stands: scots pine stands, norway spruce stands and fir stands considering different stand age classes stands stand age classes 1-20 (%) 21-40 (%) 41-60 (%) 61-80 (%) 81-100 (%) 101-120 (%) 121-140 (%) 141-160 (%) scots pine 84,8 0,6 1,7 7,0 2,2 2,8 0,6 0,3 norway spruce 2,2 1,3 38,6 45,6 11,4 0,9 fir 5,7 7,5 26,4 37,7 22,6 only one species of armillaria – a. ostoyae was found in the scots pine stands, three species: a. ostoyae, a. cepistipes and a. borealis were found in the norway spruce stands and two species: a. ostoyae and a. cepistipes were found in the fir stands (fig. 1b). in the norway spruce stands, among the identified isolates and fruit bodies belonging to genus armillaria, the species a. ostoyae attained the highest proportion – 85,5%. the remaining ones were less frequent: a. borealis – 11,4% and a. cepistipes – 3,1%. in the fir stands share of a. ostoyae was 51,2% and share of a. cepistipes was 48,8%. in the scots pine stands the species a. ostoyae was found in six forest site types, mainly in fresh mixed coniferous forest (tab. 4). in the norway spruce stands all the three identified species were found in fresh mixed broadleaved forest site type. the species a. ostoyae was recorded in six investigated site types. a. borealis was found in fresh broadleaved forest site type and fresh mixed broadleaved forest site type. a. cepistipes was found in fresh mixed broadleaved forest site type, mixed mountain forest site type, and mountain forest site type. in the fir stands a. ostoyae was noticed on three forest site types: upland forest, mountain forest and mixed mountain forest, a. cepistipes – in fresh broadleaved forest, upland forest, and mountain forest. armillaria species 215 ta b l e 4 percentage of particular armillaria species in samples from coniferous stands from different types of forest habitat type of forest habitat species of armillaria a. borealis (%) a. cepistipes (%) a. ostoyae (%) scots pine stands fcf 9,3 fmcf 78,4 mmcf 5,6 fbf 0,8 fmbf 4,8 mmbf 1,1 norway spruce stands fmcf 28,6 0,9 fbf 3,8 2,6 fmbf 96,2 12,7 muf 13,6 mmf 57,1 36,4 mf 14,3 33,8 fir stands fbf 9,5 uf 38,1 50,0 mmf 6,2 mf` 52,4 43,8 ta b l e 5 percentage of particular armillaria species in samples from coniferous stands of different stand age classes stand age classes species of armillaria a. borealis (%) a. cepistipes (%) a. ostoyae (%) scots pine stands 1-20 84,8 21-40 0,6 41-60 1,7 61-80 7,0 81-100 2,2 101-120 2,8 121-140 0,6 141-160 0,3 norway spruce stands 1-20 2,2 21-40 0,9 41-60 88,5 57,1 8,8 61-80 3,8 14,3 75,4 81-100 7,7 28,6 8,3 101-120 4,4 fir stands 41-60 9,4 61-80 9,5 6,2 81-100 23,8 28,1 101-120 42,9 31,3 121-140 23,8 25,0 216 a. żółciak a. ostoyae was found most frequently in the 1-20-year-old scots pine stands (84,8%), in the 61-80-year-old norway spruce stands (75,4%) and in the 101-120year-old fir stands (31,3%; tab. 5). a. borealis and a. cepistipes were noticed mainly in the 41-60-year-old norway spruce stands. a. cepistipes was collected particularly in the 101-120-year-old fir stands. conclusions a. ostoyae seems to be the most frequent species of armillaria in coniferous stands in poland. it was found in the all investigated stands (100% in pine, 85,5% in norway spruce and 51,2% in fir stands), in all types of forest habitat and in all stand age classes. it is in concordance with findings of many authors that a. ostoyae is responsible for most cases of armillaria attacks in conifers (g u i l l a u m i n , b e r t h e l a y 1981; r i s h b e t h 1982; r o l l -h a n s e n 1985; g u i l l a u m i n et al 1985; r y k o w s k i 1990; g u i l l a u m i n et al 1993). a. borealis was found only in the older (41-100-years-old) norway spruce stands in fertile forest site types (fbf, fmbf). a. cepistipes was noticed in the older norway spruce and fir stands and in many forest site types except for weak coniferous ones. over 50% of samples of that species were collected in the mountain sites. acknowledgement. i gratefully acknowledge dr kari korhonen from the finnish research institute for the testers of armillaria. references c a p e c k i z. 1994. rejony zdrowotności lasów zachodniej części karpat. (forest health condition re-rejony zdrowotności lasów zachodniej części karpat. (forest health condition regions in western carpathians). prace inst. bad. leśn. a, 781: 83–191. c a p e c k i z. 1997. rejonizacja zdrowotności lasów środkowej części karpat. (regions of differing health in the central part of the carpathians). prace inst. bad. leśn. a, 840: 83–191. g u i l l a u m i n j. j., b e r t h e l a y s. 1981. détermination spécifique des armillaires par la méthode des groupes de compatibilité sexuelle. spécialisation écologique des espèces françaises. agronomie 1: 897–908. g u i l l a u m i n j. j., l u n g b., r o m a g n e s i h., m a r x m ü l l e r h., l a m o u r e d., d u r r i e u g., j. j., l u n g b., r o m a g n e s i h., m a r x m ü l l e r h., l a m o u r e d., d u r r i e u g., -l u n g b., r o m a g n e s i h., m a r x m ü l l e r h., l a m o u r e d., d u r r i e u g., b., r o m a g n e s i h., m a r x m ü l l e r h., l a m o u r e d., d u r r i e u g., -r o m a g n e s i h., m a r x m ü l l e r h., l a m o u r e d., d u r r i e u g., h., m a r x m ü l l e r h., l a m o u r e d., d u r r i e u g., -m a r x m ü l l e r h., l a m o u r e d., d u r r i e u g., h., l a m o u r e d., d u r r i e u g., -l a m o u r e d., d u r r i e u g., d., d u r r i e u g., -d u r r i e u g., g., -b e r t h e l a y s., m o h a m e d c. 1985. systématique des armillaires du groupe mellea. conséquences phytopathologiques. eur. j. for. path. 15: 268–277. g u i l l a u m i n j. j., m o h a m e d c., a n s e l m i n., c o u r t e c u i s s e r., g r e g o r y o., r j. j., m o h a m e d c., a n s e l m i n., c o u r t e c u i s s e r., g r e g o r y o., r -m o h a m e d c., a n s e l m i n., c o u r t e c u i s s e r., g r e g o r y o., r c., a n s e l m i n., c o u r t e c u i s s e r., g r e g o r y o., r -a n s e l m i n., c o u r t e c u i s s e r., g r e g o r y o., r n., c o u r t e c u i s s e r., g r e g o r y o., r -c o u r t e c u i s s e r., g r e g o r y o., r r., g r e g o r y o., r -g r e g o r y o., r o., r -h o l d e n r i e d e r o., i n t i n i m., l u n g b., m a r x m ü l l e r h., m o r r i s o n d., r i s h b e t h j., te r m o r s h u i z e n a. j., t i r r ó a., va n d a m b. 1993. geographical distribution and ecology of the armillaria species in western europe. eur. j. for. path. 23: 321–341. k o r h o n e n k. 1978. interfertility and clonal size in the armillaria mellea complex. karstenia 18: 31-42. l e c h p. 2003. zagrożenie drzewostanów świerkowych w polsce przez patogeny korzeni w świetle wyników monitoringu fitopatologicznego lasów gospodarczych. (in:) a. g r z y w a c z (ed.). drzewostany świerkowe – stan, problemy, perspektywy rozwojowe. materiały sesji naukowej ptl, ustrońjaszowiec: 92–107. m a ń k a k. 1953. badania terenowe i laboratoryjne nad opieńką miodową – armillaria mellea (vahl) quél. prace inst. bad. leśn. 94: 1–96. m a ń k a k. 1998. fitopatologia leśna. pwril. warszawa. 368 pp. m a ń k a m., p r z y b y ł k., m a ł e c k a m. 2003. choroby świerka na tle zmian środowiska. (in:) materiałach sesji naukowej ptl, drzewostany świerkowe: stan, problemy, perspektywy rozwojowe. ustroń-jaszowiec: 63–76. r i s h b e t h j. 1982. species of armillaria in southern england. pl. path. 31: 9–17. r o l l -h a n s e n f. 1985. the armillaria species in europe. eur. j. for. path. 15: 22–31. armillaria species 217 r o m a g n e s i h., m a r x m ü l l e r h. 1983. etude complémentaire sur les armillaires annelées. bull. soc. mycolog. de france 99: 301– 324. r y k o w s k i k. 1985. niektóre troficzne uwarunkowania patogeniczności armillaria mellea (vahl) quèl. w uprawach sosnowych. prace inst. bad. leśn. 640: 1–140. r y k o w s k i k. 1990. opieńkowa zgnilizna korzeni. pwril. warszawa. 16 pp. s i e r o t a z. 2001. choroby lasu. centrum informacyjne lasów państwowych. 156 pp. tw a r o w s k a i. 1965. opieńka miodowa. pwril warszawa. 54 pp. tw a r o w s k i z., tw a r o w s k a i. 1959. studies and observations on armillaria mellea (vahl) quél. as the cause of mass dying-off of forest stands. prace inst. bad. leśn. 192: 1–61. ż ó ł c i a k a. 1999. identyfikacja gatunków grzybów z rodzaju armillaria (fr.: fr.) staude w polsce. prace inst. bad. leśn. 888: 3–19. opieńki w drzewostanach iglastych s t r e s z c z e n i e materiał badawczy (owocniki, fragmenty drewna przerośniętego grzybnią, ryzomorfy) pobierano w różnych latach, na jednorazowych powierzchniach w wybranych drzewostanach iglastych: sosnowych, świerkowych i jodłowych. izolaty identyfikowano za pomocą testów intersterylności. owocniki identyfikowano na podstawie cech makroskopowych i mikroskopowych. wśród badanych próbek stwierdzono trzy gatunki opieniek. stwierdzono w drzewostanach: sosnowych – a. ostoyae, świerkowych – a. ostoyae, a. borealis i a. cepistipes, jodłowych – a. ostoyae i a. cepistipes. a. ostoyae wydaje się być jednym z najczęściej występujących gatunków opieniek w drzewostanach iglastych w polsce. stwierdzono ten gatunek we wszystkich badanych drzewostanach iglastych. jego udział stanowił 100% prób zebranych w drzewostanach sosnowych, 85,5% – w drzewostanach świerkowych oraz 51,5% – w drzewostanach jodłowych. a. ostoyae stwierdzono na wszystkich typach siedliskowych lasu i w drzewostanach we wszystkich klasach wieku. a. borealis stwierdzono tylko w drzewostanach świerkowych, w wieku 41-100 lat, na bogatych siedliskach. a. cepistipes stwierdzono w drzewostanach świerkowych i jodłowych, w starszym wieku, na większości badanych siedlisk. 2014-01-01t11:46:21+0100 polish botanical society impact of light on yielding of some pleurotus sp. strains marek siwulski, mirosława ziombra and krzysztof sobieralski department of vegetable crops, poznań university of life sciences dąbrowskiego 159, pl-60-594 poznań, fungus@up.poznan.pl siwulski m., ziombra m., sobieralski k.: impact of light on yielding of some pleurotus sp. strains. acta mycol. 47 (1): 65–73, 2012. light is an important factor deciding about yielding and morphological characters of pleurotus carpophores. the objective of the research was to ascertain the impact of period and intensity of lighting on yielding and carpophore morphological features of four strains of oyster mushroom. the following strains were investigated: p. ostreatus: px, k22 and p80 strains, p. pulmonarius: p20 strain. fluorescent lamps with day-light were used to provide light in the cultivation room. the following lighting periods were used: 6, 10 and 14 hours/ day and the applied lighting intensity included: 100, 300, 500 and 700 lx. lighting exerted a significant impact on yielding. the highest carpophore crop was recorded when the applied lighting intensity was 500 and 700 lx for the period of 14 h/d. the highest mean mass of carpophores was recorded at 14-hour light exposure and 500 and 700 lx lighting intensity. carpophore morphological features modified by the lighting period and its intensity included the cap diameter as well as the length and thickness of the stem. key words: oyster mushroom, cultivation, lighting intensity, carpophore, morphological traits introduction species from the pleurotus genus, including their strains, differ with regard to characters of their carpophores (ziombra, gembiak 2000; siwulski et al. 2006). yields of mushrooms from the pleurotus genus depend on many factors; apart from genetic properties, also environmental factors play a significant role in this regard (lelley 1991; shah et al. 2004). the mycelium of mushrooms from the pleurotus genus does not require light for its growth (sharma 2004), nevertheless, light is necessary for the proper development of carpophores (olivier 1988; royse, zaki 1991). trukhonovets (1991) maintains that during the period of carpophore development and growth, light is an important factor deciding about yielding and morphological characters of fruiting acta mycologica vol. 47 (1): 65–73 2012 66 m. siwulski et al. bodies. experiments conducted by the above-mentioned researcher showed that light quantities required for carpophore development can be regulated by shortening the exposure time and increasing the light intensity or, conversely, by lengthening the time of exposure to light and decreasing its intensity. in other words, the total quantity of light is decisive for normal development of carpophores. it should, however, be emphasised that there are significant differences between strains. selection of profusely fruiting strains and providing optimal conditions for their cultivation are commonly considered to be among key yield-forming factors. due to considerable variability of the pleurotus genus, both with regard to morphological as well as functional features, many forms of this mushroom have already been selected and are used in cultivation as separate strains. strains differ among one another with regard to the weight of their carpophores, size and thickness of the pileus as well as the length and thickness of the stem. these traits alter depending on cultivation conditions, although they remain characteristic for the given strain (curvetto et al. 2002; siwulski et al. 2006). the objective of the presented research project was to ascertain the impact of light on yielding and carpophore morphological features of four strains of oyster mushroom. material and methods the following species and strains of oyster mushroom were investigated: pleurotus ostreatus jacq.ex.(fr.) kumm. – px, k22 and p80 strains, pleurotus pulmonarius (fr.) quel. – p20 strain. the evaluation of yields depending on the duration and intensity of lighting exposure was carried out in air-conditioned chambers of the department of vegetable crops, university of life sciences in poznań. the experiment was set up in a random design in four replications and two cultivation cycles. the cultivation substrate was wheat straw cut into chaff of 3-5 cm length. the substrate of approximately 70% moisture content was pasteurised at the temperature of 60°c for the period of 48 hours. after the pasteurisation process, the substrate was cooled down to the temperature of 25°c and mixed with oyster mushroom mycelium. the oyster mushroom mycelium as propagation material was produced on wheat grain. the proportion of the applied mycelium in relation to the cultivation substrate was established at 5%. the substrate, together with oyster mushroom mycelium, were placed in perforated polyethylene bags; 10 dm3 in each bag. the process of overgrowing of the oyster mushroom mycelium through the substrate took place in darkness in a cultivation facility in which the temperature was maintained at the level of 18-20°c and air humidity – at 80-85%. once the substrate was overgrown by the mycelium of the examined strains, different lighting conditions were applied for the cropping period. fluorescent lamps with light similar to natural light (day-light) were used to provide light in the facilities. the following lighting periods were used: 6, 10 and 14 hours/day and the applied lighting intensity included: 100, 300, 500 and 700 lx. measurements of the impact of light on yielding of some pleurotus sp. strains 67 lighting intensity were performed on the substrate surface with the assistance of a luxmeter l-20. the yield mass in relation to the substrate dry matter was determined and biometric measurements of fruiting bodies were taken. a sample for biometric measurements consisting of 40 carpophores was collected randomly from each experimental combination. the diameter and thickness of the cap and length and thickness of the stem were determined. the results comprising yields and morphological traits of carpophores were analysed for mean values from replications and cultivation cycles. the analysis of variance for three-factorial experiments was performed calculating lsd at the significance level of α=0.05. results lighting exerted a significant impact on yielding. the highest carpophore crop, irrespective of the strain, was recorded in combinations where the applied lighting intensity was 500 and 700 lx for the period of 14 h/d. the experimental strains responded differently to the applied light regimes. at lighting intensity of 500 and 700 lx, the highest and non-significantly differing crops were recorded for px, k22 and p80 strains, whereas p20 strain exhibited the weakest response to changes in the lighting intensity, especially at 14-hour light exposure (tab. 1). the light exposure of 6 h/d, irrespective of the lighting intensity, resulted in a significant decline of yields in comparison with 10 and 14 h/d lighting period. the highest yields were obtained at 14-hour light exposure. apart from the duration of the light exposure, also light intensity played a significant role. the highest crops were recorded at 500 and 700 lx lighting intensity. yields obtained in such conditions did not differ significantly irrespective of the period of lighting. table 1 pleurotus yield in relation to intensity and period of lighting (g x kg-1 d.m. of substrate) lighting strain (h) (lx) px p20 k22 b80 6 100 300 500 700 188 206 278 346 205 275 290 330 122 138 245 282 182 235 285 296 mean 256 275 196 250 10 100 300 500 700 228 380 512 522 402 518 622 635 232 326 480 492 330 462 634 780 mean 411 544 382 552 14 100 300 500 700 426 595 805 830 532 660 668 672 284 398 615 728 325 554 890 912 mean 664 633 506 670 lsd 0.05 for strain=60, for lighting intensity=86, for period of lighting=82, for interaction=115 68 m. siwulski et al. it was demonstrated on the basis of analyses of lighting intensity and time interrelationships that it was not possible to shorten the time of light exposure even when light intensity was increased up to 700 lx. on the other hand, it is not advisable to reduce lighting intensity below 500 lx, if we want to lengthen the duration of light exposure at the expense of lighting intensity. the examined strains responded similarly to lighting duration in the 24-hour period. the recorded mean carpophore mass characteristic for a given strain changed following different light exposure of the cultivation. when a 14-hour lighting regime was employed, the mean weight of harvested carpophores was higher in comparison with fruiting bodies grown in 6 or 10-hour regimes. the mean carpophore weight increased together with the length of the lighting period per day. also lighting intensity exerted influence on carpophore weight. when the applied lighting intensity was 100 or 300 lx, carpophores of significantly smaller weight were obtained than at table 2 carpophore mean mass of pleurotus strains in relation to intensity and period of lighting (g) lighting strain (h) (lx) px p20 k22 b80 6 100 300 500 700 22 28 36 42 28 32 32 32 20 20 22 24 26 28 40 42 mean 32 31 22 34 10 100 300 500 700 38 42 48 50 34 36 38 38 24 36 38 38 32 44 46 48 mean 45 37 34 43 14 100 300 500 700 40 58 66 68 24 36 48 50 24 34 48 50 36 50 58 60 mean 58 40 39 51 lsd 0.05 for strain=8, for lighting intensity=8, for period of lighting=10, for interaction=15 table 3 cap diameter of pleurotus strains in relation to intensity and period of lighting (mm) lighting strain (h) (lx) px p20 k22 b80 6 100 300 500 700 22 30 36 34 30 32 36 38 18 20 26 26 22 22 28 32 mean 31 34 23 26 10 100 300 500 700 32 42 50 53 42 52 52 52 24 32 37 38 28 38 46 46 mean 44 50 33 40 14 100 300 500 700 38 56 72 74 46 48 52 52 28 38 47 52 32 46 58 62 mean 60 50 41 50 lsd 0.05 for strain=6, for lighting intensity=8, for period of lighting=6, for interaction=11 impact of light on yielding of some pleurotus sp. strains 69 the same period to light exposure but of 500 and 700 lx. however, lighting intensity of 500 and 700 lx applied only for 6 or 10 hours exerted a negative influence on carpophore weight. the highest weight of fruiting bodies was recorded at 14-hour light exposure and 500 and 700 lx lighting intensity (tab. 2). strain p20 was characterised by the weakest response to changes in lighting conditions. the remaining strains were found to respond similarly to changes in both lighting intensity and duration. cap diameter and thickness were features characteristic for a given strain and, similarly to the carpophore mass, depended on lighting intensity and duration. carpophores with the largest and thickest caps were observed in combinations with 14 h/d light exposure and 500 and 700 lx lighting intensity (tabs 3 and 4). the applied lighting intensity of cultivations affected the length and diameter of mushroom stems (tabs 5 and 6). fruiting bodies with the shortest stems were found in strains cultivated for the longest light exposure (14 h/d). in addition, a distinct table 4 cap thickness of pleurotus strains in relation to intensity and period of lighting (mm) lighting strain (h) (lx) px p20 k22 b80 6 100 300 500 700 8 10 12 12 6 6 8 8 6 6 6 6 8 8 10 10 mean 11 7 6 9 10 100 300 500 700 12 12 14 14 8 9 9 9 8 8 10 10 12 12 14 14 mean 13 9 9 13 14 100 300 500 700 12 15 17 17 8 10 10 10 10 10 12 12 12 15 16 16 mean 15 10 11 15 lsd 0.05 for strain=2, for lighting intensity=2, for period of lighting=2, for interaction=2 table 5 stem length of pleurotus strains in relation to intensity and period of lighting (mm) lighting strain (h) (lx) px p20 k22 b80 6 100 300 500 700 40 38 36 36 14 14 12 12 38 38 34 34 38 36 34 34 mean 38 13 36 35 10 100 300 500 700 40 36 34 32 14 12 12 10 36 34 33 33 34 32 28 26 mean 36 12 34 30 14 100 300 500 700 36 32 28 26 12 10 10 10 34 26 25 25 34 28 26 26 mean 31 11 28 28 lsd 0.05 for strain=6, for lighting intensity=8, for period of lighting=3, for interaction=10 70 m. siwulski et al. tendency was found with decreasing lighting intensity for the development of carpophores with increasingly long stems. the stem thickness of carpophores declined with decreasing lighting intensity and shortening of time exposure to light. the proportion of stems in the weight of carpophores ranged from 11 to 42% depending on strain, lighting intensity and light exposure time. p20 strain was characterised by the smallest proportion (11-16%) of stems in carpophores; short stems are typical for this strain and in comparison with other strains, the length of their stems is least dependent on variations in lighting. in the case of the remaining strains, the percentage proportion of the stem weight in the carpophore weight was considerably higher ranging from 25 to 42%, depending on the lighting regime. in general, it can be said that the better the lighting conditions, the smaller was the proportion of the stem weight in the carpophore weight. it is true that when lighting conditions table 6 stem diameter of pleurotus strains in relation to intensity and period of lighting (mm) lighting strain (h) (lx) px p20 k22 b80 6 100 300 500 700 8 8 8 9 8 8 10 10 8 8 9 10 9 9 11 11 mean 8 9 9 10 10 100 300 500 700 9 10 10 12 10 10 10 10 8 9 10 10 8 8 10 10 mean 10 10 9 9 14 100 300 500 700 10 12 14 14 10 12 12 12 8 8 10 10 8 8 10 10 mean 13 12 9 9 lsd 0.05 for strain=2, for lighting intensity=3, for period of lighting=2, for interaction=3 table 7 proportion of stem in carpophore mass of pleurotus strains in relation to intensity and period of lighting (%) lighting strain (h) (lx) px p20 k22 b80 6 100 300 500 700 42 38 36 36 16 16 14 14 36 36 32 32 40 36 34 30 mean 38 15 34 35 10 100 300 500 700 38 35 30 28 15 14 12 12 36 36 30 30 34 32 30 30 mean 33 13 33 32 14 100 300 500 700 33 30 27 27 13 12 11 11 32 30 27 27 28 26 26 25 mean 29 12 29 26 lsd 0.05 for strain=5, for lighting intensity=5, for period of lighting=4, for interaction=8 impact of light on yielding of some pleurotus sp. strains 71 deteriorated, the stem also became thinner but it also became longer and this caused that the stem weight increased affecting its percentage proportion in the carpophore weight (tab. 7). discussion light, along with other external factors, exerts a significant impact on the growth and development processes of carpophores of mushrooms from the pleurotus genus. it acts as a signal triggering off various biophysical and biochemical processes ultimately leading to morphological and phototrophic reactions (trukhonovets 1991). in the performed investigations, the size and thickness of the cap and length and thickness of the carpophore stem characteristic for the strain altered under the influence of the applied various lighting regimes. the mean carpophore weight was a trait characteristic for a given strain but dependent on time and intensity of lighting of cultivations. also trukhonovets (1991) reported a dependence of the size of the cap and stem length on lighting intensity. among important characters affecting salability of fruiting bodies is the ratio of the cap size to the stem size. stems of carpophores of the majority of oyster mushroom species, with the exception of pleurotus eryngii, are inedible and should be as small as possible. in our investigations, irrespective of the applied lighting regime, the shortest stems were determined in the case of carpophores of the p20 strain, whereas in the remaining examined strains stems were longer and changed depending on lighting intensity. however, according to literature data, there is no agreement as to the optimal range of lighting intensity recommended in the mushroom cultivation of pleurotus genus and, depending on individual researchers, this range fluctuates from 300 to 1500 lx (lelley 1991; stamets 2000; oei 2003; ziombra et al. 2008). in the presented experiments, the optimal lighting intensity to obtain high crops of advantageous carpophore morphological features ranged from 500 to 700 lx for px, k22 and b80 strains. the examined p20 strain was characterized by the lowest requirements regarding the applied lighting regimes and gave similarly high yields both at 300 and 700 lx lighting intensity. carpophore morphological traits of the p20 strain remained unchanged throughout the examined range of lighting intensity. the appropriate growth of fruiting bodies is also affected by the length of the lighting period in the 24 h rhythm (trukhonovets 1991). the research results obtained in this study indicate that the applied 14 h lighting regime turned out to be the most favourable for carpophore development. crops harvested following the 8 h light regime were lower than in the case of 10 h and 14 h light exposure despite the application of higher lighting intensity of 500 and 700 lx. recapitulating, it was concluded on the basis of the performed experiments that cultivations of the examined species of mushrooms from the pleurotus genus should receive, during the period of the development and growth of fruiting bodies not less than 10 hours of light per 24 h. in order to obtain carpophores of large caps and short stems, the lighting intensity should not be lower than 500 lx. in addition, the 72 m. siwulski et al. obtained research results showed that a longer lighting period within the 24 h period failed to compensate insufficient lighting intensity and, conversely, despite high intensity of lighting, shortening of the lighting exposure time below 10 hours/day is not recommendable. conclusions 1. the highest yields of px, k22 and b80 strains of oyster mushroom were obtained from cultivations exposed to 10 and 14 hours of light per 24 h applying 500 and 700 lx lighting intensity. 2. the p20 strain of oyster mushroom gave similar yields at lighting intensity ranging from 300 through 500 up to 700 lx. 3. the examined oyster mushroom strains developed carpophores with largest caps within the range of lighting considered as optimal to obtain abundant crops. 4. carpophore morphological features modified by the length of the lighting period and its intensity included the cap diameter as well as the length and thickness of the stem. references curvetto n. r., figlas d., devalis r., delmastro s. 2002. growth and productivity of different pleurotus ostreatus strains on sunflower seed hulls supplemented with n-nh4 + and/or mn (ii). bioresource technology 84(2): 171–176. lelley j. 1991. pilzanbau. biotechnologie der kulturspeisepilze. ulmer, stuttgart, 404 pp. oei p. 2003. mushroom cultivation. backhuys publishers leiden, netherlands, 429 pp. olivier j. m. 1988. les besoins en lumiere dans la culture des pleurotes. bull. fed. nat. syndicate agricol. cult. champignons 40: 1433–1439. royse d. i., zaki s. a. 1991. yield stimulation of pleurotus sp. by dual nutrient supplementation of pasteurized wheat straw. (in:) m.j. maher (ed.). science and cultivation of edible fungi. balkema, rotterdam: 545–547. shah z. a., ashraf m., ishtiag m. 2004. comparative study on cultivatin and yield performance of oyster mushroom (pleurotus ostreatus) on different substrates. pakistan j. nutrition 3 (3): 158–160. sharma b. b. 2004. effect of duration of light on radial growth of pink oyster mushroom. indian pathology 57 (2): 234. siwulski m., sobieralski k., ławicka k. 2006. porównanie plonowania wybranych odmian i krzyżówek boczniaka pleurotus sp. folia horticulturae, supl. 2: 130–133. stamets p. 2000. growing gourmet and medicinal mushrooms. ten speed press, berkeley, pp.574. tan q., wang zq., cheng jh., guo q., guo l. 2005. cultivation of pleurotus spp. in china. acta edulis fungi 12: 338–342. trukhonovets v. v. 1991. effect of illumination intensity on the formation of fruiting bodies in pleurotus ostreatus (jacq. fr.) kumm. ukr. bot. zh. 48 (2): 67–72. ziombra m., gembiak r. 2000. cechy morfologiczne owocników grzybów z rodzaju pleurotus sp. rocz. ar pozn. 31: 573–577. ziombra m., czerwińska-nowak a., ławicka k. 2008. wpływ natężenia światła i czasu oświetlenia na plon i cechy morfologiczne owocników kilku odmian boczniaka. zeszyty problemowe postępów nauk rolniczych 527: 335–341. impact of light on yielding of some pleurotus sp. strains 73 wpływ światła na plonowanie kilku ras pleurotus sp. streszczenie światło jest ważnym czynnikiem decydującym o plonowaniu i cechach morfologicznych owocników boczniaka. celem badań było określenie wpływu czasu i natężenia oświetlenia na wielkość plonu oraz cechy morfologiczne owocników czterech ras boczniaka. przedmiotem badań były rasy pleurotus ostreatus: px, k22 i p80 oraz rasa pleurotus pulmonarius:p20. do oświetlenia pomieszczeń uprawowych użyto lamp fluoroscencyjnych o świetle zbliżonym do naturalnego. okres oświetlenia wynosił 6, 10 i 14 godzin na dobę. zastosowano oświetlenie o natężeniu 100, 300, 500 i 700 lx. stwierdzono, że światło wywierało znaczący wpływ na plonowanie. największe plony owocników uzyskano stosując oświetlenie o natężeniu 500 i 700 lx przez 14 godzin na dobę. cechami morfologicznymi owocników modyfikowanymi przez długość okresu oświetlenia i jego intensywność były średnica kapelusza oraz długość i grubość trzonu. 2014-01-02t12:03:35+0100 polish botanical society phyllosphere mycobiota on garden pond plants maria kowalik department of plant protection, hugo kołłątaj university of agriculture in kraków 29 listopada 54, pl-31-425 kraków, m.kowalik@ogr.ur.krakow.pl kowalik m.: phyllosphere mycobiota on garden ponds plants. acta mycol. 47 (1): 11–19, 2012. investigations were conducted on calamus, common cattail, soft rush, yellow iris and white water lily plants in twenty ponds in malopolska and podkarpacie regions. mycobiota existing in the phyllosphere caused discolouring and necroses of leaves and shoots. 88 species of mycobiota were identified and isolated from the diseased tissues. dominant were alternaria alternata, epicoccum nigrum and isaria farinosa. fungi of genera: aspergillus, botrytis, chaetomium, cladosporium, fusarium, ilyonectria, mortierella, mucor, penicillium, phialophora, phoma, pleustomophora, sordaria, trichoderma and umbelopsis were also numerous. the monophagous and the polyphagous were identified. key words: garden pond, water plants, phyllosphere, fungi, chromistan fungi, plant diseases introduction garden pond is a specific system functioning on the border of water and land ecosystem. plant phyllosphere is a natural place of mycobiota existence. in a pond environment mycobiota pressure on various water plant species is heightened, which results in the occurrence of disease symptoms. the problems of water plants’ health state, as results from the literature of the subject, has been still a rarely addressed issue, therefore presented investigations aimed at determining the species composition of the mycobiota settling the phyllosphere of ponds and causing diseases visible as colour changes and necrosis. presented work is a recapitulation of research results showing taxonomic and species diversity of the mycobiota existing in the phyllosphere of five plants: calamus, common cattail, soft rush, yellow iris and white water lily – invariable components of pond ecosystem (kowalik, maik 2010; kowalik 2011a, b; kowalik, cwynar 2011; kowalik 2012). acta mycologica vol. 47 (1): 11–19 2012 12 m. kowalik materials and methods field investigations were conducted during four vegetation seasons (2006, 20082010) in twenty ponds situated in house gardens in the malopolska and podkarpacie regions. the investigations covered fourty plants from each species: calamus acorus calamus l., common cattail typha latifolia l., soft rush juncus effusus l., yellow iris iris pseudoacorus l. and white water lily nymphaea alba l. each year, five observation were carried out from april to october at four-six-weekly long intervals. mycobiota were determined on the basis of etiological symptoms and microscopic analyses of mycelium, sporodochia, pycnidia, conidophores and conidia forming on diseased plants. 3200 infected fragments of necrotic plant tissues, including 600 fragments of calamus, soft rush and common cattail, each and 700 fragments of yellow iris and white water lily, each were collected for mycological analyses. isolation and culturing of mycobiota were conducted using the standard methods practised in mycology. mycobiota were classified to appropriate species using mycological keys: booth (1971); batko (1975); domsch et al. (1980), sutton (1980); ho (1981); ellis and ellis (1987); ramirez (1982); sivenesan (1984). results in result of mycological analysis of the diseased tissues 2675 colonies of fungi and fungi-like organisms chromista were obtained, which belong to the following genera: alternaria, aspergillus, botrytis, chaetomium, cladosporium, davidiella, drechslera, epicoccum, fusarium, humicola, ilyonectria, isaria, khuskia, mortierella, mucor, penicillium, phaeosphaeria, phialophora, phoma, phyllosticta, phytophthora, rhizopus, sordaria, trichoderma, umbelopsis and other (tab. 1). the mycobiota were represented by 88 species, which evidences species diversity in pond phyllosphere. the least number of species – 27 settled calamus and the highest – 43 common cattail. 37 were isolated from yellow iris tissues, 34 from soft rush, whereas 40 fungi species and 2 species of fungus-like organisms were isolated from white water lily. the following were prevailing in the identified mycobiota community: alternaria alternata constituting 33.23% of the whole community, epicoccum nigrum – 10.24% and isaria farinosa – 9.35%. in the influent group (making up from 1 to 5% of the mycobiota community) identified were: aspergillus niger f. niger, botrytis cinerea, chaetomium globosum, cladosporium cladosporioides, ilyonectria radicicola, mortierella alpina, mucor hiemalis, penicillium aurantiogriseum, p. expansum, p. hirsutum var. hirsutum, p. verrucosum, phoma medicaginis var. medicaginis, pleustomophora richardsiae, sordaria fimicola and umbelopsis isabellina. polyphagous species, such as: a. alternata, a. niger. f. niger, e. nigrum, fusarium poae, i. farinosa, m. alpina, s. fimicola and u. isabellina were identified among fungi existing in water plant phyllosphere. these fungi were settling necrotic tissues of all five plant species. b. cinerea, ch. globosum, c. cladosporioides, p. aurantiogriseum, p. phyllosphere mycobiota on garden pond plants 13 expansum, p. verrucosum, phialophora cyclaminis, ph. medicaginis var. medicaginis, pleustomophora richardsiae and rhizopus stolonifer existed on four plant species in ponds. the monophagous were less numerous: ascochyta acori was isolated from calamus, phaeosphaeria eustoma (syn. leptosphaeria typhae), ph. typharum (syn. l. typharum) and periconia typhicola from common cattail, whereas physalospora scirpi (syn. arthrinium curvatum v. minus), phaeosphaeria juncina (syn. leptosphaeria juncina), morenoina paludosa, paraphaeosphaeria michotii and septoriella junci from soft rush. phytophthora citricola and ph. undulata chromistan fungi were isolated from diseased petioles of white water lily. disease symptoms were visible on pond plants in all years of the investigations. broad spots, yellow in colour visible on calamus in may and june were located in top leaf parts, in the late summer season the colour changed to brownish-brown. the border of spots was changing colour from yellow through reddish to light brown. the spots were situated all over the leaf surface. another symptom were narrow spindleshaped brown smudges and small, round dots with red-russet rim located along the vein. tissue in places of the spot underwent necrosis and the heaviest intensity of necroses occurred in october. the following were most frequently isolated from the diseased tissues: a. alternata, i. farinosa, e. nigrum, b. cinerea and a. niger f. niger. common cattail plants revealed first disease symptoms in may, visible as yellowish discolorations on top and base of leaves. in september the symptoms were visible on the surface of leaf blades as brown-red spots and smudges. by the end of vegetation necrotic spots covered the whole leaf blade area. beside a. alternata, i. farinosa and e. nigrum, the most numerous fungi isolated from the necrotized tissues included p. expansum, s. fimicola and p. richardsiae. the first symptoms of leaf spot disease were visible on soft rush in april as delicate chlorotic discolourings in leaf top parts. with time the leaves were yellowing and browning. streaks of chlorotic tissue changing its colour to brown-black were noticeable on stems. these symptoms sometimes led to dieback of plant clusters in a pond. the species dominant on necrotised soft rush tissues were also a. alternata, i. farinosa and e. nigrum, b. cinerea, m. alpina and s. fimicola were numerous, too. disease symptoms and their exacerbation on yellow iris leaves were diversified in individual vegetation periods. generally, first disease symptoms were observed in may and the greatest intensification occurred in september or october. small elongated or oval yellowish spots were visible on leaf blade, which in time changed their colour to orange or brown. the spots were surrounded by a clear reddish or brown rim. initially top leaf parts were dying back, whereas by the end of vegetation a full necrosis of leaf blades occurred. spots and necrosis of yellow iris leaves were most frequently caused by: a. alternata, e. nigrum, i. farinosa, ph. medicaginis var. medicaginis and phyllosticta pseudoacori. leaf spot diseases, called brown leaf spot disease was visible on white water lily every year and in all ponds. the symptoms of disease, visible as tiny chlorotic discolourings of leaf blade, appeared in may. the symptoms were intensifying gradually and by the end of vegetation brown spots surrounded with yellowish or reddish rim covered a considerable part of leaf. necrotic tissue was covered by a coating of a concentrically zoned mycelium. the tissue was crumbled and leaf perforation 14 m. kowalik table1 mycobiota isolated from affected tissues of garden pond plants fungus c al am us a co ru s ca la m us c om m on ca ta il ty ph a la tif ol ia so ft r us h ju nc us e ff us us y el lo w ir is ir is ps eu do ac or us w hi te w at er lil yn ym ph ae a al ba to ta l pe rc en ta ge [% ] alternaria alternata (fr.) keissl. 277 183 110 215 104 889 33.23 arthrinium sphaerospermum fuckel 1 1 0.04 arthrinium sporophleum kunze 5 5 0.19 arthrinium urticae m. b. ellis 1 1 0.04 arthroderma tuberculatum kuehn 1 1 0.04 ascochyta acori oudem. 3 3 0.11 aspergillus alutaceus berk. et m. a. curtis 1 1 0.04 aspergillus nidulans (eidam) g. winter var. nidulans 5 5 0.19 aspergillus niger f. niger tiegh. 20 16 9 2 7 54 2.02 aspergillus versicolor (vuill.) tirab. 6 7 8 21 0.79 boeremia exigua var. exigua (desm.) aveskamp, gruyter et varkley 1 1 2 0.07 boeremia hedericola (durieu et mont.) aveskamp, gruyter et verkley 1 6 7 0.26 botryotinia convoluta (drayton) whetzel 6 6 0.22 botrytis cinerea pers. 23 22 10 2 57 2.13 chaetomium cochlioides palliser 1 3 4 0.15 chaetomium elatum kunze 1 7 8 0.30 chaetomium globosum kunze 23 14 6 3 46 1.72 cladosporium cladosporioides (fresen) g. a. de vries 1 13 1 12 27 1.01 cladosporium herbarum (pers.) link 1 1 0.04 cladosporium sphaerospermum penz. 4 9 13 0.49 clonostachys rosea f. rosea (link) schroers, samuels, seifert et w. gams 5 5 0.19 coleophoma empetri (rostr.) petr. 1 2 6 9 0.34 davidiella macrocarpa crous, k. schub. et u. braun 7 6 13 0.49 diplodia melaena lév. 3 3 0.11 drechslera poae (baudys) shoemaker 16 16 0.60 epicoccum nigrum link 33 108 31 90 12 274 10,24 fusarium culmorum (w. g. sm.) sacc. 9 4 1 14 0.52 fusarium oxysporum schltdl. 17 17 0.64 fusarium poae (peck) wollenw. 4 1 4 2 7 18 0.67 fusarium sporotrichioides sherb. 9 2 11 0.41 gibberella tricincta el-gholl, mcritchie, schoult et ridings 4 1 5 0.19 humicola fuscoatra traaen 3 4 5 12 0.45 humicola grisea traaen var. grisea 2 7 9 0.34 ilyonectria radicicola (gerlach et l. nilsson) cheverri et c. g. salgado 3 28 31 1.16 isaria farinosa (holmsk.) fr. 38 44 82 45 41 250 9.35 khuskia oryzae h. j. huds. 8 3 11 0.41 mammaria echinobotryoides ces. 1 2 3 0.11 morenoina paludosa j. p. ellis 6 6 0.22 mortierella alpina peyronel 11 4 19 1 10 45 1.68 mortierella parvispora linnem. 2 1 3 0.11 mucor hiemalis wehmer 1 9 22 32 1.20 mycosphaerella iridis (auersw.) j. schröt. 2 2 0.07 myrothecium cinctum (corda) sacc. 10 10 0.37 phyllosphere mycobiota on garden pond plants 15 neottiospora caricina (desm.) höhnel 3 3 0.11 nowakowskiella elegans (nowak.) schröt. 1 1 0.04 paecilomyces variotii bainier 1 1 0.04 paraconiothyrium minitans (w. a. campb.) verkley 2 2 0.07 paraphaeosphaeria michotii (westend.) o. erikss. 1 1 0.04 paraphoma chrysanthemicola (hollós) gruyter, aveskamp et varkley 3 2 5 0.19 penicillium aurantiogriseum dierckx 16 9 3 8 36 1.35 penicillium citrinum thom 6 2 8 0.30 penicillium expansum link 19 34 1 32 86 3.21 penicillium hirsutum var. hirsutum dierckx 15 10 15 40 1.50 penicillium lanosum westling 12 12 0.45 penicillium verrucosum dierckx 12 19 1 79 111 4.15 penicillium waksmanii k. m. zalessky 1 12 13 0.49 periconia typhicola e. w. mason et m. b. ellis 2 2 0.07 pestalotiopsis sydowiana (bres) b. sutton 1 1 0.04 phaeosphaeria eustoma (fuckel) l. holm 1 1 0.04 phaeosphaeria juncina (auersw.) l. holm 4 4 0.15 phaeosphaeria typharum (desm.) l. holm 4 4 0.15 phialophora cinerescens (wollenw.) j. f. h. beyma 10 2 12 0.45 phialophora cyclaminis j. f. h. beyma 3 5 10 4 22 0.82 phoma leveillei boerema et g. j. bollen var. leveillei 2 17 19 0.71 phoma medicaginis var. medicaginis malbr. et roum. 7 7 5 24 43 1.61 phyllosticta pseudacori (brunaud) allesch. 11 11 0.41 physalospora scirpi (gutner) arx 6 5 11 0.41 phytophthora citricola savada 3 3 0.11 phytophthora undulata (h. e. petersen) m. w. dick 12 12 0.45 pleustomophora richardsiae (nannf.) l. hostert, w. gams et crous 21 3 6 4 34 1.27 rhizopus arrhizus a. fisch. var. arrhizus 10 10 0.37 rhizopus stolonifer (ehrenb.) vuill. 9 6 1 1 17 0.64 scopulariopsis koningii (oudem.) vuill. 1 1 0.04 septoriella junci (desm.) b. sutton 1 9 10 0.37 sordaria fimicola (roberge et desm.) ces et de not. 5 25 18 9 24 81 3.03 talaromyces wortmannii c. r. benj. 7 7 0.26 thielavia terricola (j. g. gilman et e. v. abbott) c. w. emmons 1 1 0.04 trichoderma aureoviride rifai 16 16 0.60 trichoderma harzianum rifai 5 2 7 0.26 trichoderma koningii oudem. 2 2 0.07 trichoderma pseudokoningii rifai 14 14 0.52 trichoderma viride pers. 8 8 0.30 trichothecium roseum (pers.) link 3 2 5 0.19 ulocladium botrytis preuss 2 2 0.07 umbelopsis isabellina (oudem.) w. gams 2 3 5 9 15 34 1.27 umbelopsis nana (linnem.) arx 4 4 0.15 volutella arundinis desm. 7 7 0.26 total 565 577 430 530 573 2675 100.00 table1– cont. 16 m. kowalik happened. the most numerous species in the community of isolated fungi were: a. alternata, p. verrucosum, i. farinosa, p. expansum and i. radicicola. in two ponds ph. citricola and ph. undulata were isolated from white water lily tissues. the symptoms on rhizomes, petioles and leaf blades pointed to an occurrence of a disease determined as a black rot on rhizomes. discussion a comparison of results of mycobiota isolation from phyllosphere of plants which are the fixed components of the analyzed ponds and information about the occurrence of diseases on their aboveground and underground organs (westcott 1971; van der aa 1978; sutton 1980; ellis, ellis 1987; helberg 1998; wagner 2000; kowalik, krasny 2009; mazurkiewicz-zapałowicz, grajewski 2010), revealed the most serious pathogen threat from the polyphagous, such as: a. alternata, b. cinerea, e. nigrum, s. fimicola. these fungi, most numerously isolated by the end of vegetation period colonized plant tissues causing necrosis. the fungi constantly accompanying pond plants included also those from mortierella and umbelopsis genera, identified as hydrophilous (domsch et al. 1980) and fungi from aspergillus, penicillium, isaria, and phialophora genera existing in many ecological niches (kowalik, maik 2010; kowalik 2011b; kowalik, cwynar 2011). slightly less numerous cladosporium spp. isolated from the phyllosphere are regarded as causes of leaf spot disease, necroses and disturbances in many plants growth, not only in ponds (westcott 1971; van der aa 1978; pląskowska 2010; kowalik 2011b). a fact of settling the petioles and leaf blades by fusarium spp. is noticeable in the presented investigations. these fungi, especially f. culmorum, f. poae and f. sporotrichioides, which beside a. alternata, a. niger f. niger, p. aurantiogriseum, p. hirsutum var. hirsutum, and p. verrucosum are counted to toxicogenous ones which intensify necrotization process considerably contributing to fast destruction of plant tissues (płażek 2011; chełkowski 2012). fungi from trichoderma genus, which are well known for their antagonism towards pathogenic fungi from fusarium and ilyonectria genera, were relatively little numerous on the plant tissues (except for white water lily). intensified occurrence of necrotrophs from alternaria, sordaria, epicoccum, chaetomium, mucor and rhizopus genera was noticed by the end of vegetation period, (in october), which confirms their ability to develop in dead tissues of the diseased plants. biodiversity of mycobiota in plant phyllosphere is evidenced by isolation of incidental saprotrophs which may develop both on live and dead tissue. arthrinium, boeremia, diplodia, mammaria, nowakowskiella, paraconiothyrium, scopulariopsis, thielavia, ulocladium and other fungi only occasionally present on diseased shoots and leaves may be also counted to this group. phyllosphere mycobiota on garden pond plants 17 isolating from water plants the monophagous, which are strict parasites infesting calamus and soft rush in rush communities, adamska 2005; mazurkiewiczzapałowicz et al. 2006; mazurkiewicz-zapałowicz, grajewski 2010) and wild sedge, testifies their expansiveness and proves that they sporulate both on wild plants and ornamental cultivars (czerniawska, adamska 2009). in ponds particular attention should be paid to hazard provided by phytophthora, because chromistan fungi which belong to these genera commonly colonize perennial plants, dwarf shrubs, coniferous and deciduous plants in their natural environment, whereas water is the agent spreading propagule (orlikowski 2006). the hazard has been confirmed by isolation of ph. citricola and ph. undulata from while water lily plants. it should be surmised that the composition of mycobiota settling the phylosphere of plants depended on plant physiological properties and active substance content (ethereal oils, alkaloids, phytoncides and dyes), therefore the least number of fungi existed on calamus and soft rush. the role of fungi accompanying calamus vegetation and reducing the content of active substance were described by mazurkiewiczzapałowicz and grajewski (2010). while comparing the frequency of diseases occurrence on pond plants, it was noticed that while water lily and common cattail plants were the most frequently infested, which confirmed previous reports (kowalik, krasny 2009). assessing the healthiness of all plants planted in ponds it should be emphasized that it was satisfactory, which does not change the fact that visible leaf spot diseases and necroses on leaves and shoots do not improve aesthetic values of a pond, on the contrary, they greatly diminish them. conclusions 1. dominance of mycobiota of alternaria, botrytis, epicoccum, fusarium, isaria, and penicillium genera in the phyllosphere of pond plants: white water lily, yellow iris, common cattail, soft rush and calamus results from their common occurrence in the environment, ability for fast spreading on plants growing in close vicinity, abilities for facultative parasitism and considerable reproductive potential. 2. it was stated that mycobiota of botrytis, ilyonectria, fusarium, phialophora, pleustomophora, phoma and phytophthora accompanying the plants in a pond during vegetation season caused diverse discolourings and necroses on leaves and shoots. 3. intensified tissue necrotization process on all plants in ponds occurred by the end of vegetation process and was connected with the frequency of occurrence in the phylosphere of necrotrophs of alternaria, sordaria, epicoccum, chaetomium, mucor and rhizopus genera. 4. while causing necroses, phyllosphere mycobiota of pond plants contributed to a destruction of tissues, which resulted in a diminishing of aesthetic values of ponds. 18 m. kowalik references adamska i. 2005. fungal species new in poland on carex and juncus. acta mycol. 40: 19–24. batko a. 1975. zarys hydromikologii. pwn. booth c. 1971. the genus fusarium. cmi, kew, surrey, england. chełkowski j. 2012. mikotoksyny, grzyby toksynotwórcze i mikotoksykozy. www.cropnet.pl/ mycotoxin: 1-100. [dostęp 18.01.2012]. czerniawska b., adamska i. 2009. grzyby pasożytnicze występujące na turzycach. prog. plant protection/ post. ochr. roślin 49 (1): 187–190. domsch k.h., gams w., anderson t.h. 1980. compendium of soil fungi. acad. press. london, new york, toronto, sydney, san francisco. ellis m. b., ellis j.p. 1987. microfungi of land plants. croom helm. london, sydney. helberg t. 1998. ogrody wodne. delta w-z. warszawa. ho h.h. 1981. synoptic keys to the species of phytophthora. mycologia 73: 705–714. kowalik m. 2011a. fungi associated with plants calamus acorus calamus l. during the vegetation season. phytopathologia 60: 29–33. kowalik m. 2011b. grzyby towarzyszące roślinom pałki szerokolistnej typha latifolia l. w okresie wegetacji. prog. plant protection/post. ochr. roślin 51 (1): 269–273. kowalik m. 2012. fungi and chromistan fungi associated with plants white water lily nympaea alba l. during the vegetation season. acta scientarum polonorum, seria hortorum cultus (in print). kowalik m., cwynar a. 2011. grzyby towarzyszące roślinom situ rozpierzchłego juncus effusus l. w okresie wegetacji. prog. plant protection/post. ochr. roślin 51 (2): 652–655. kowalik m., krasny m. 2009. fungi occurring on garden pond plants. phytopathologia 51: 21–26. kowalik m., maik m. 2010. grzyby w fylosferze irysa żółtego iris pseudoacorus l. prog. plant protection/ post. ochr. roślin 50 (1): 218–221. mazurkiewicz-zapałowicz k., grajewski j. 2010. grzyby fyllosfery acorus calamus l. – cennej rośliny leczniczej. prog. plant protection/post. ochr. roślin 50 (1): 236–239. mazurkiewicz-zapałowicz k., wróbel m., silicki a., wolska m. 2006. studies on phytopathogenic and saprotrophic fungi in rush association of lake glinno (nw poland). acta mycol. 41(1): 125–138. orlikowski l.b. 2006. relationship between source of water used for plant sprinkling and occurrence of phytophthora shoot rot and tip blight in container ornamental nurseries. j. plant prot. res. 46: 163–168. pląskowska e. (ed.). 2010. miejskie tereny zielone – zagrożenia. uwp. wrocław: 1–165. płażek a. 2011. patofizjologia roślin. wydawnictwo uniwersytetu rolniczego w krakowie, kraków. ramirez c. 1982. manual and atlas of the penicillia. elsevier biomedical press. amsterdam, new york, oxford. sivenesan a. 1984. the bitunicate ascomycetes and their anamorphs. j. cramer, vaduz. sutton b.c. 1980. the coelomycetes – fungi imperfecti with pycnidia, acervuli and stromata. cmi, kew, surrey. van der aa h.a. 1978. a leaf spot disease of nymphaea alba l. in the netherlands. e. j. plant pathology 84: 109–115. wagner a. 2000. fungi isolated from leaves of different iris spp. rocz. ar poznań cccxxi, ogrodn. 30: 165–169. westcott c.1971. plant disease handbook. van nostrand reinhold co. new york, cincinnati, toronto, london, melbourne. phyllosphere mycobiota on garden pond plants 19 fyllosferowe mykobiota roślin oczek wodnych streszczenie w badaniach terenowych i laboratoryjnych przeprowadzonych w latach 2006, 2008-2010, w dwudziestu oczkach wodnych usytuowanych na terenie małopolski i podkarpacia określono stan zdrowotny: tataraku zwyczajnego acorus calamus l., pałki szerokolistnej typha latifolia l., situ rozpierzchłego juncus effusus l., irysa żółtego iris pseudoacorus l. i grzybienia białego nymphaea alba l. mykobiota bytujące w fyllosferze roślin powodowały różnorakie przebarwienia i nekrozy. z porażonych tkanek roślin wyodrębniono 2675 kolonii mykobiota, należących do 88 gatunków. w zbiorowisku wyodrębnionych mykobiota dominowały: alternaria alternata, epicoccum nigrum i isaria farinosa. mniej licznie izolowano grzyby z rodzajów: aspergillus, botrytis, chaetomium, cladosporium, fusarium, ilyonectria, mortierella, mucor, penicillium, phialophora, phoma, pleustomophora, sordaria, trichoderma i umbelopsis. fyllosferowe mykobiota roślin oczek wodnych powodując nekrozy przyczyniały się do destrukcji tkanek, co skutkowało obniżeniem walorów dekoracyjnych oczek wodnych. 2014-01-02t11:58:26+0100 polish botanical society fungi and fungi-like oomycetes isolated from affected leaves of rhododendron maria kowalik department of plant protection, university of agriculture in kraków al. 29. listopada 54, pl-31-425 kraków, mkowalik@ogr.ar.krakow.pl kowalik m.: fungi and fungi-like oomycetes isolated from affected leaves of rhododendron. acta mycol. 43 (1): 21–27, 2008. the aim of the work is to identify fungi and fungi-like oomycetes occurring on affected leaves of rhododendron rhododendron l. mycological analyses were carried out on 200 leaves collected from green areas of kraków from may till september 2005. isolated fungilike oomycetes belonged to 67 taxa. the most frequently found fungi included: alternaria alternata, aspergillus niger, botrytis cinerea, coelophoma empetri, nigrospora sphaerica, pestalotia sydowiana, phialophora cyclaminis, phomopsis archeri, septoria azalea and sordaria fimicola. among fungi-like organisms phytophthora cinnamomi and p. citricola were isolated. key words: rhododendron, leaves, isolation, fungi-like oomycetes introduction rhododendrons growing in urban green areas are susceptible to various pathogenes. infected plants develop discolouration, brown spots and necroses, affecting their aesthetic value. the symptoms of infection, observable from spring to autumn, increase when the plants are in bloom resulting in dieback and leaf drop. the damage is caused by fungi-like oomycetes of genera pythium and phytophthora and fungi of genera: botrytis, cercospora, colletotrichum, cylindrocladium, exobasidium, microsphaera, ovulinia, pestalotia, phomopsis, phyllosticta, pycnostysanus, ramularia, rhizoctonia and septoria (farr et al. ����� �er�er� ��a��a ����� �erre� ����� �r������ �er�er� ��a��a ����� �erre� ����� �rliko��ki ����� łaba�o��ki et al. 2001; kita, mazurek 2003; kowalik et al. 2006; kowalik, muras 2007). the aim of this work was to identify the organisms occurring on affected rhododendron leaves in green areas of kraków. acta mycologica vol. 43 (1): 21–27 2008 22 m. kowalik material and methods leaves of rhododendron with the symptoms of discoloration, brown spots or ne-rhododendron with the symptoms of discoloration, brown spots or necroses were collected from the plants in the botanical garden of the jagiellonian university, the zoological garden and town lawns in kraków. 200 leaves from 50 plants were collected for mycological analysis. the numbers of leaves taken from the top part (t), central part (c) and lower part (l) of the plant were approximately equal. the leaf samples were disinfected in 70% ethyl alcohol and 5 parts of each leaf from the area between ill and healthy tissue were put on petri dishes with 2% pda medium. the colonies of fungi and fungi-like oomycetes �ere ide�tified u�i�g variou� media for ide�tificatio�: guba (����)� dom�ch et al. (����)� sutto� (����)� ho (1981), ellis and ellis (1987). results from the collected material 721 colonies of fungi, 19 colonies of fungi-like oomycetes and 150 colonies of bacteria (not being the subject of this paper) were isolated. the isolated fungi and oomycetes belonged to 67 taxa within 37 genera. among the dominat fungi the following constitued more than 66 per cent of the total: alternaria alternata, aspergillus niger, botrytis cinerea, cladosporium macrocarpum, coelophoma empetri, humicola grisea v. grisea, nigrospora sphaerica, pestalotia sydowiana, phialophora cyclaminis, ph. richardsiae, phoma fimeti, ph. macrostoma, phomopsis archeri, pycnostysanus azaleae, septoria azalea and sordaria fimicola. less frequent were: acremonium furcatum, cladosporium sphaerospermum, colletotrichum gloeosporioides, cylindrocladium scoparium, epicoccum purpurascens, exobasidium vaccinii, humicola fuscoatra v. fuscoatra, phoma chrysanthemicola and ph. putaminum. fungi belonging to 20 taxa occurred once or twice. fungi-like oomycetes were represented by phytophthora cinnamomi and ph. citricola (tab. 1). among the species isolated from the leaves situated in the central part of the plants, the following were predominant: a. alternata, b. cinerea, c. empetri, c. gloeosporioides, e. purpurascens, n. sphaerica, p. sydowiana, s. fimicola and fungi belonging to genera: aspergillus, chaetomium, cladosporium, humicola and penicillium. from leaves situated in the lower parts of the plant, the following were isolated: cylindrocarpon destructans, c. scoparium, ph. archeri, rhizoctonia solani, s. fimicola, trichothecium roseum and fungi of genera: acremonium, fusarium, mortierella and phialophora. from the lower parts also ph. cinnamomi and ph. citricola were isolated. from the leaves situated at the top part of the plant, numerous colonies of a. alternata, c. empetri, s. azalea, s. fimicola were isolated. less frequent were: e. vaccinii, mucor hiemalis, p. sydowiana, pycnostysanus azaleae and fungi of genera trichoderma and phoma. the frequency of species occurrence of fungi and fungi-like oomycetes on leafblades was differentiated. the following species occurred at the highest frequency: a. alternata, s. fimicola, p. sydowiana, s. azalea and c. empetri. they were isolated (respectively) from 27, 11, 9, 8 and 7 leaves. less frequently (on 5-6 leaves) occurred: aspergillus niger, b. cinerea, cladosporium macrocarpum, c. gloeosporioides, fungi and fungi-like 23 ta b l e 1 fungi and fungi-like oomycetes isolated from affected leaves of rhododendron species frequency of occurrence percentage of occurrence the number of leaves the fungus species was isolated from localization of leaves* acremonium butyri (van beyma) w. gams 6 0.81 2 l acremonium furcatum f. et v. moreau ex w. gams 8 1.08 2 l acremonium fusidioides (nicot) w. gams 1 0.14 1 c acremonium kiliense grütz 4 0.54 1 l alternaria alternata (fr.) keissler 131 17.70 27 tcl arthrinium pheosphermum (corda) m. b. ellis 2 0.27 1 c aspergillus niger van tiegh. 21 2.84 6 c aspergillus wentii wehmer 4 0.54 1 c botrytis cinerea pers. ex nocca et balb. 21 2.84 6 c chaetomium globosum kunze ex steud. 3 0.41 1 c chaetomium indicum corda 2 0.27 1 c chrysosporium asperatum carm. 6 0.81 2 c chrysosporium merdarium (link ex grev.) carm. 1 0.14 1 l cladosporium cladosporioides (fres.) de vries 6 0.81 2 c cladosporium macrocarpum preuss 19 2.57 6 c cladosporium sphaerospermum penz. 10 1.35 2 c coelophoma empetri (rostr.) petrak 25 3.38 7 tc colletotrichum gloeosporioides (penz.) sacc. 14 1.89 5 c coniothyrium minitans w. a. campbell 6 0.81 2 c cylindrocarpon destructans (zins.) scholten 6 0.81 2 l cylindrocarpon tenue bugn. 2 0.27 1 c cylindrocladium scoparium morg. 8 1.08 2 l doratomyces stemonitis (pers. ex steud.) morton 2 0.27 1 t epicoccum purpurascens ehrenb. ex schlecht. 12 1.62 4 c exobasidium vaccinii (fuckel) woron. 12 1.62 3 t fusarium culmorum (w. g. sm.) sacc. 5 0.68 1 l fusarium graminearum schwabe 2 0.27 1 l fusarium stilboides wollenw. 2 0.27 1 l gloeosporium rhododendri briosi et cav. 2 0.27 1 c humicola fuscoatra traaen var. fuscoatra 8 1.08 4 c humicola grisea traaen var. grisea 17 2.30 5 c mammaria echinobotryoides ces. 7 0.95 2 c morenoina rhododendri j. p. ellis 2 0.27 1 c mortierella alpina peyronel 2 0.27 1 l mortierella horticola linnem. 1 0.14 1 l mortierella isabellina oudem. 1 0.14 1 l mortierella parvispora linnem. 4 0.54 1 l mucor hiemalis wehmer f. hiemalis 4 0.54 1 t myrothecium cinctum (corda) sacc. 1 0.14 1 c nectria inventa pethybr. 2 0.27 1 c nigrospora sphaerica (sacc.) mason 21 2.84 5 c penicillium expansum link ex gray 7 0.95 2 c pestalotia sydowiana bres. 40 5.41 9 tc phialophora cyclaminis van beyma 23 3.11 5 l phialophora richardsiae (nannf.) conant 16 2.16 4 l phoma chrysanthemicola hollòs 11 1.49 3 t phoma epicoccina punithalingam, tulloch et leach 7 0.95 2 t phoma eupyrena sacc. 6 0.81 2 t 24 m. kowalik humicola grisea v. grisea, n. sphaerica, ph. cyclaminis, ph. fimeti, ph. macrostoma, ph. archeri and p. azaleae. on 3-4 leaves occurred e. purpurascens, e. vaccinii, h. fuscoatra v. fuscoatra, ph. richardsiae, ph. chrysanthemicola and ph. cinnamomi. the remaining were isolated form 1-2 leaves (tab. 1). rhododendron symptoms caused by fungi and fungi-like oomycetes were varied. from chlorosis affected leaves with discoloration changes alongside the main veins, c. destructans, c. tenue, c. scoparium, and also three species of fusarium were isolated. fungus r. solani, causing the rhizoctoniosis in rhododendron, was isolated from the leaves with irregular brown-grey spots, surrounded by a large yellow area. b. cinerea, the cause of botrytis petal blight, was isolated from rapidly dying leaves, with the symptoms of necrosis, such as large brown spots covered with dense grey mould. fungus e. vaccinii, was isolated from deformed leaves with small reddish and brownish galls, typical of azalea leaf a�d flo�er gall. the cause of large oval necrotic �pot� �ith dark li�i�g� a�d vi�ible pic�idia �a� ide�tified a� ph. archeri. other pathogens occurring in phyllosphere, such as c. gloeosporioides (causing rhododendron anthracnose), gloeosporium rhododendri, morenoina rhododendri, c. empetri and s. azalea (the reason for leaf scorch) caused round, oval or irregular necrotic spots of different shapes. a sign of c. gloeosporioides infection was grey spots, with visible rings and picnidia. in case of s. azalea, small oval, grey-brown spots with darker lining were seen. necrotic spots caused by ph. cyclaminis and p. richardsiae were small irregular with uneven edges. p. sydowiana was isolated from large light brown necrotic spots appearing on the edges and the tops of the leaves. leaf petioles and blades, with brown discolouration and necroses forming a cline �hape� �ere the reflectio� of ph. cinnamomi and ph. citricola presence. these pathogens were also isolated from fast expanding brown oval spots along the leaf margin. phoma fimeti brun. 18 2.43 6 t phoma herbarum westd. 3 0.41 1 t phoma leveillei boerema et bollen 1 0.14 1 t phoma macrostoma mont. 17 2.30 6 t phoma medicaginis malbr. et roum. 5 0.68 1 t phoma pomorum thüm. 5 0.68 1 t phoma putaminum speg. 9 1.22 2 t phomopsis archeri sutton 23 3.11 6 l phytophthora cinnamomi rands 12 1.62 3 l phytophthora citricola sawada 7 0.95 2 l pseudeurotium zonatum van beyma 2 0.27 1 c pycnostysanus azaleae (peck) e. mason 16 2.16 5 t rhizoctonia solani kühn 2 0.27 1 l septoria azalea voglino 33 4.46 8 t sordaria fimicola (rob.) ces. et de not. 53 7.16 11 tcl trichoderma harzianum rifai 2 0.27 1 t trichoderma pseudokoningii rifai 2 0.27 1 t trichoderma viride pers. ex gray 4 0.54 1 t trichothecium roseum (pers.) link ex gray 3 0.41 1 l total 740 100.00 abbreviations: * – localization of leaves: t – top part of the plant; c – central part of the plant; l – lower part of the plant. tab. 1 cont. fungi and fungi-like 25 discussion different discolorations, brown spots and necroses, visible on rhododendron leave� i� gree� area� of �rakó�� �ere cau�ed by ma�y orga�i�m� cla��ified a� fungi and �omycete�. accordi�g to farr et al. (����)� �er�er a�d ��a��a (����)� �erre� et al. (����)� łaba�o��ki et al. (����) rhodode�dro� leave� ca� be i�fected by fungi and fungi-like organisms of different genera including: botrytis, cercospora, colletotrichum, cylindrocladium, exobasidium, gloeosporium, macrophoma, microsphaera, monochaetia, pestalotia, pestalotiopsis, phomopsis, phyllosticta, phytophthora, septoria, rhizoctonia and others. while kita and mazurek (2003) isolated fungi of genera: alternaria, botrytis, cladosporium, epicoccum, fusarium, humicola, mucor, penicillium, pestalotia, phoma, rhizopus, sclerotinia and trichoderma from the phyllosphere of rhododendron and azalea growing in the botanical garde� a�d arboretum. thi� re�earch �ork co�firm� that most of fungi and fungilike oomycetes isolated from the affected leaves of rhododendron belong to the mentioned above taxa. the determination of organisms inhabiting affected rhododendron leaves enabled to show the relation between pathogens and pathologic symptoms. the isolation of fungi such as: c. destructans, c. tenue, c. scoparium in the leaves with the discolouratio� alo�g�ide the mai� vei� i�dicate� that the root �y�tem of the pla�t� �a� fir�ty attacked� cau�i�g root rot� a� me�tio�ed by �er�er (���5) a�d łabanowski et al. (2001). also in the work by kowalik and muras (2007) fungi c. destructans, c. scoparium and numerous species of fusarium were numerously isolated from dropped rhododendron leaves. fungus r. solani was the reason for rhododendron rhizoctoniosis, b. cinerea caused grey mould, c. gloeosporioides – anthracnose, and s. azalea – rhodode�dro� leaf �corch (łabanowski et al. 2001), while fungus e. vaccinii spending �i�ter i� flo�er a�d leaf bud� of rhodode�dro��� i� �pri�g cau�ed the di�ea�e called azalea leaf a�d flo�er gall (orlikowski 1999). the occurrence in phyllosphere of pathogens such as c. gloeosporioides, g. rhododendri, m. rhododendri, c. empetri and s. azalea (numerously isolated in this reasearch work) and leaf symptoms caused by them were documented in the papers by werner and ��a��a (����)� łaba�o��ki et al. (2001), kita and mazurek (2003). in this work p. sydowiana was frequently isolated. łaba�o��ki et al. (2001) do not attribute necroses to this species. the role of p. sydowiana in causing necroses of heathers in nurseries and in permanent sites was described by kowalik and sa-heathers in nurseries and in permanent sites was described by kowalik and sagan (2005) as well as kowalik and wandzel (2005), while kita and mazurek (2003), in phyllosphere of azaleas and rhododendrons found quite numerously occurring: pestalotia fibricola, p. rhododendrii and p. truncata. in the works by kowalik and muras (2007) fungus p. sydowiana together with a. alternata, occurred causing spots and necroses of rhododendron leaves and intensifying the process of leaf drop were isolated at large frequency. the description of phytophtorosis symptoms on leaves and petioles of rhodode�dro�� a�d their effect� i� rhodode�dro� pla�ti�g� are co�firmed i� the �ork� by łaba�o��ki et al. (����)� �erre� (����)� �er�er (���5)� �o�alik et al. (����). the occurrence of ph. citricola in rhododendron nurseries and susceptibility of many rhododendron taxa to this pathogen is described by orlikowski and szkuta (2003), 26 m. kowalik while orlikowski and szkuta (2002) highlight the role ph. cinnamomi in causing phytophtorosis on container-grown ericaceous plants in ornamental nurseries. while analysing the plant material, it was found out that, in addition to the above mentioned causes of discolorations, brown spots and necroses, the secondarily i�vadi�g fu�gi o� pathological leave� �ere ide�tified. they �ere the follo�i�g: a. alternata, s. fimicola, n. sphaerica, a. niger and the species of genera: acremonium, cladosporium, humicola, mortierella, mucor, penicillium, phoma and trichoderma. accordi�g to �er�er a�d ��a��a (����) the�e fu�gi ca� i�te��ify pathogenic processes. inhabiting small necrotic areas in a short time they rapidly inten-in a short time they rapidly intensify the process of necrosis. a large participation of fungi from genera: alternaria, cladosporium, epicoccum and phoma in the phyllosphere of rhododendrons is mentioned by kita and mazurek (2003). their work indicates that these saprotrophs dominate in the structure of the populations of isolated fungi. the obtained results co�firm co��iderable participation of a. alternata, e. purpurascens, c. macrocarpum, c. sphaerospermum and many species of phoma in the populations of fungi isolated from necrotic tissues of rhododendrons. it can be concluded that a large biodiversity of fungi and fungi-like oomycetes isolated from the affected rhododendron leaves in the green areas of kraków caused different pathologic symptoms and decreased decorative value of the plants. conclusions the most numerous fungi occurring in the rhododendron plantings in the green 1. areas of kraków were: alternaria alternata, aspergillus niger, botrytis cinerea, coelophoma empetri, nigrospora sphaerica, pestalotia sydowiana, phialophora cyclaminis, phomopsis archeri, septoria azalea and sordaria fimicola among the fungi-like oomycetes2. phytophthora cinnamomi and ph. citricola were isolated from the affected rhododendron leaves. the causes of discolorations, brown spots and necroses of the rhododen-, brown spots and necroses of the rhododen-spots and necroses of the rhododenand necroses of the rhododen-and necroses of the rhododen-3. dron leaves were the pathogens belonging to genera: botrytis, colletotrichum, cylindrocarpon, cylindrocladium, exobasidium, fusarium, pestalotia, phomopsis, phytophthora, rhizoctonia and septoria. commonly occurring saprotrophs of genera: 4. acremonium, alternaria, cladosporium, coelophoma, epicoccum, humicola, nigrospora, phoma, sordaria a�d other�� i�te��ified the proce�� of leaf �ecro�i�. on the rhododendron plants the symptoms of phytophthorosis, grey mould, 5. rhizoctoniosis, anthracnose, leaf scorch, azalea leaf a�d flo�er gall were found. the effect was leaf drop and, in consequence decreased decorative value of the plants. references domsch k. h., gams w., anderson t. h. 1980. compendium of soil fungi. acad. press. london, new york, toronto, sydney, san francisco. ellis m. b., ellis j. p. 1987. microfungi on land plants. croom helm. london, sydney. farr d., esteban h. b., palm m. e. 1996. fungi on rhododendron: a world reference. parkway publish., bonne, nc. guba e. f. 1961. monograph of monochaetia and pestalotia. harvard univ. press, cambrige. ho h. h. 1981. synoptic keys to the species of phytophthora. mycologia 73: 705–714. fungi and fungi-like 27 �ita �.� mazurek j. ���3. skład gatu�ko�y fyllo�fery róża�ecz�ikó� � �grodzie bota�icz�ym �e �rocła�iu i � arboretum � �oj�ła�icach. erica polo�ica �4: �5–3�. kowalik m., sagan a. 2005. fungi causing dying out heather in permanent plantings. acta mycol. 40 (2): 191–195. �o�alik m.� �a�dzel a. ���5. grzyby po�odujące zamiera�ie �adzo�ek �rzo�u. acta agrobot. 5� (�): 237–242. �o�alik m.� mura� p. ���7. grzyby za�iedlające opadłe li�cie róża�ecz�ika. rocz. ar � poz�a�iu 3�3. ogrodnictwo 41: 69–73. �o�alik m.� �li�a a.� paliga a. ����. choroby róża�ecz�ikó� (rhododendron l.) na terenach zieleni krakowa. erica polonica 17: 33–41. łaba�o��ki g.� �rliko��ki l.� soika g.� �ojdyła a.� �orbi� m. ����. �chro�a ro�li� �rzo�o�atych. plantpress. �rliko��ki l. b. ����. po�łocz�ik azalio�y �a azalii i róża�ecz�iku. �chro�a ro�li� �:��. orlikowski l. b., szkuta g. 2002. occurrence of phytophthora cinnamomi on ericaceous plants in container-grown ornamental nurseries in poland. j. plant protect. res. 42 (2): 157–163. orlikowski l. b., szkuta g. 2003. phytophthora citricola on rhododendron spp. in polish nurseries. j. plant protect. res. 43 (1): 19–24. sutton b. c. 1980. the coelomycetes. cmi, kew, surrey. werner m. 2005. grzyby za�iedlające korze�ie róża�ecz�ikó� � �zkółkach i ich z�acze�ie dla zdro�ot�o�ci ro�li�. erica polo�ica ��: �5–7�. �er�er m.� ��a��a h. ����. choroby pędó� i li�ci róża�ecz�ika. �chro�a ro�li� �: ��–��. werres s. 2000. phytophthora spp. an rhododendron und azaleen. (in:) j.westhoff (ed.). rhododendron und immergrüne laubgehölze. jahrbuch der deutschen rhododendron gesselschaft 1999: 99–113. grzyby i organizmy grzybopodobne oomycetes wyizolowane z porażo�ych li�ci róża�ecz�ika s t r e s z c z e n i e celem pracy była ide�tyfikacja grzybó� i orga�izmó� grzybopodob�ych �omycete� �y�tępujących �a porażo�ych li�ciach róża�ecz�ika rhododendron l. materiał do badań �ta�o�iło ��� li�ci (z obja�ami przebar�ień� plami�to�ci i �ekroz) zebra�ych z 5� krze�ó� �a terenie ogrodu botanicznego uj, ogrodu zoologicznego oraz na skwerkach miejskich krako�a. �yodręb�io�o grzyby i orga�izmy grzybopodob�e �omycete� (� liczbie 74� kolo�ii)� �ależące do �7 tak�o�ó�. na porażo�ych li�ciach �ajlicz�iej �y�tępo�ały grzyby: alternaria alternata, aspergillus niger, botrytis cinerea, coelophoma empetri, nigrospora sphaerica, pestalotia sydowiana, phialophora cyclaminis, phomopsis archeri, septoria azalea i sordaria fimicola. spo�ród orga�izmó� grzybopodob�ych� z porażo�ych li�ci róża�ecz�ika �yizolo�a�o phytophthora cinnamomi i ph. citricola. spra�cami przebar�ień� plami�to�ci i �ekroz �a li�ciach róża�ecz�ika były patoge�y z rodzajó�: botrytis, colletotrichum, cylindrocarpon, cylindrocladium, exobasidium, fusarium, pestalotia, phomopsis, phytophthora, rhizoctonia i septoria. licz�ie �y�tępujące �aprotrofy z rodzajó�: acremonium, alternaria, cladosporium, coelophoma, epicoccum, humicola, nigrospora, phoma, sordaria i i��e i�te��yfiko�ały proce� �ekrotyzacji li�ci. na róża�ecz�ikach rozpoz�a�o obja�y fytoftorozy� �zarej ple��i� rizokto�iozy� a�trak�ozy� �eptoriozy� po�łocz�ika azalio�ego� � �y�iku których dochodziło do zamiera�ia i opada�ia li�ci� a tym �amym ob�iże�ia �aloró� dekoracyj�ych ro�li�. 2014-01-01t11:47:00+0100 polish botanical society some additional laboulbeniales (ascomycetes) new to poland tomasz majewski department of plant pathology, warsaw agricultural university nowoursynowska 166, pl 02 787 warszawa tomasz_majewski@sggw.pl m a j e w s k i t.: some additional laboulbeniales (ascomycetes) new to poland. acta mycol. 41(1): 65 72, 2006. six fungal species belonging to the order laboulbeniales new to poland are described and illustrated: dimeromyces corynetis thaxter, euphoriomyces magnicellulatus santamaria, laboulbenia cornuta thaxter, l. manubriolata thaxter, l. nana sugiyama and l. olisthopi spegazzini. the status of the latter species, also reported from ukraine, is briefly discussed. key words: laboulbeniales, dimeromyces, euphoriomyces, laboulbenia, poland, ukraine introduction six species of the order laboulbeniales new to poland are described and illustrated. recorded only recently, they are not listed in the author’s previous publications (m a j e w s k i 1994, 1999). two species were collected during field studies (laboulbenia cornuta and l. manubriolata). the others were identified on insects in the following entomological collections: private collection of roman królik, kluczbork (dimeromyces corynetis), private collection of rafał ruta, piła (euphoriomyces magnicellulatus), collection of the museum and institute of zoology, polish academy of sciences, łomna (laboulbenia nana), and the natural history museum, institute of systematics and evolution of animals, polish academy of sciences, kraków (laboulbenia olisthopi). additionally, laboulbenia elaphricola j.siemaszko et w.siemaszko should also be classified in the mycoflora of poland. the species was considered to be a synonym of laboulbenia elaphri spegazzini by the author (m a j e w s k i 1994). d e k e s e l and k r a s t i n a d e k e s e l (2006), however, have now confirmed specific distinctness of laboulbenia elaphricola. a total of 206 species of the order laboulbeniales are now known from poland. they are presented in a forthcoming publication (m a j e w s k i ms.) which provides a full list of references and the descriptions of the localities in poland. acta mycologica vol. 41 (1): 65-72 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 66 t. majewski aknowlogments. the author would like to thank mr roman k r ó l i k and mr rafał ruta as well as the collection curators in łomna and kraków, dr. violetta to m a s z e w s k a and dr. daniel k u b i s z , for kindly providing insects for examination. the specimens listed in this study are in the author’s collection which will be handed over to the herbarium of the institute of botany, polish academy of sciences. descriptions of species dimeromyces corynetis thaxter dimeromyces corynetis thaxter, proc. amer. acad. arts 48: 157. 1912 (‘d. corynitis’). female thallus yellowish, 290 μm long. receptacle 50 μm long (without the foot), consisting of four cells; the basal cell large, elongated, the second one flattened, bearing laterally a short, slender secondary appendage; the third cell isodiametric, bearing laterally the perithecium, and the fourth cell isodiametric, bearing laterally a long, slender, 6-celled secondary appendage. primary appendage being a continuation of the receptacle axis, unicellular, elongated, as long as the basal cell of the receptacle. perithecium fusiform, 175 x 45 μm, with indistinct neck, stalk cell 75 μm long. male thallus not found (fig. 1a). on necrobia ruficollis (fabricius) (coleoptera, cleridae): wierzbica górna, kluczbork poviat, ex cadaver of capreolus capreolus (l.), leg. roman k r ó l i k , 28.8.1990, tm 10100 (host ex coll. r. k r ó l i k ). only one complete thallus was found on the host’s elytron. its features are consistent with the descriptions and figures given by t h a x t e r (1924) and s a n t a m a r i a (2003). dimeromyces corynetis, an infrequent species, parasitizes representatives of the genera corynetes and necrobia. it is known from america (argentina and hawaii) and from western europe, namely from spain, great britain, france, and italy (s a n t a m a r i a et al. 1991; s a n t a m a r i a 2003). euphoriomyces magnicellulatus santamaria euphoriomyces magnicellulatus santamaria, revista iberoam. micol. 8: 48. 1991. thallus hyaline, 85-135 μm long. receptacle consisting of 6-8 superposed cells, 40-63 μm long. the basal cell usually somewhat elongated, the remaining cells rather flattened in the lower part of the receptacle and isodiametric in its upper part. except the basal one, the receptacle cells cut off bilaterally basal cells of short secondary antheridial blanchlets or stalk cells of perithecia. primary appendage is a prolongation of the receptacle, not exceeding the perithecial apex. it consists of one or two cells bearing antheridia laterally and distally. perithecia one or two per thallus, 50-75 x 18-25 μm, elongated, broadest near the middle, with indistinct neck. fig. 1: b-e. on hydnobius multistriatus (gyllenhall) (coleoptera, leiodidae): pieskowa skała, kraków poviat, 18.8.2001, leg. rafał r u t a , tm 10512–10514 (host ex coll. r. r u t a ). many thalli were found on the elytra of one insect in an entomological collection. they are mostly consistent with the description given by s a n t a m a r i a (1991, 2003); however, thalli with two mature or maturing perithecia are frequent. euphoriomyces magnicellulatus seems to be a very rare species. it has been reported only from spain on hydnobius spinipes (gyllenhal) (type host), leiodes sp. and baptolinus affinis (paykull), and from the balearic islands on hydnobius sp. some additional laboulbeniales (ascomycetes) new to poland 67 fig. 1. dimeromyces corynetis thaxter. a female thallus (drawn from tm. 10100). eupho riomyces magnicellulatus santamaria. b paired thalli with one perithecium (tm 10513), c e thalli with two perithecia (tm 10514). laboulbenia manubriolata thaxter. f mature thallus (tm 10161). scale bar: 100 μm. 68 t. majewski (s a n t a m a r i a 1991, 2003). the above hosts belong to the family leiodidae. the polish host, hydnobius multistriatus, is a rare species, and its lifestyle and ecological preferences are not known (b u r a k o w s k i et al. 1978). laboulbenia cornuta thaxter laboulbenia cornuta thaxter, proc. amer. acad. arts sci. 30: 476. 1895. thalli similar to those of laboulbenia luxurians (see m a j e w s k i 1994). their particularly characteristic features include flattened cell iii situated obliquely, large cell v, and hyaline, short anterior appendage branchlets directed towards the perithecium. l. cornuta differs from l. luxurians in having larger and almost isodiametric basal cells of appendage as well as a unicellular straight horn on the perithecial apex directed anteriorly. thalli 192-220 μm long (including horn), receptacle 105-120 μm long, perithecium (without horn) 88-95 x 38-48 μm (fig. 2 a, b). on bembidion obliquum (sturm) (coleoptera, carabidae, subgenus notaphus): zielona góra: jędrzychów, sand by the water on the bottom of a large sand pit surrounded by a pinus sylvestris l. forest by the s town border, 14.8.2002, leg. t. m a j e w s k i , tm 9420. the thalli of l. cornuta discussed here are consistent with t h a x t e r ’s description (1895, 1896) and figures given by t h a x t e r (1896) and s a n t a m a r i a (2001). laboulbenia cornuta is a rare species. it was described from the u.s.a. (t h a x t e r 1895), and was later reported from cuba (b a l a z u c 1977), hungary (b á n h e g y i 1944) and spain (s a n t a m a r i a 2001). hosts are beetles of the genus bembidion sensu lato belonging to the subgenus notaphus stephens, which is often considered to be a separate genus in the entomological literature. laboulbenia manubriolata thaxter laboulbenia manubriolata thaxter, proc. amer. acad. arts sci. 51: 44. 1915. thallus yellowish, slender, 245 μm long. cells i, ii, iii and iv elongated, cell v half as long as cell iv. cell wall of cell ii and upper half of cell i distinctly tuberculate, somewhat darker than the other part of the thallus. insertion cell thin, constricted. the outer basal cell of appendage forming blunt outgrowth, appendage branchlets not preserved in the described thallus. cells vi and vii elongated, perithecium 75 x 28 μm, subfusiform, more than one half free, with indistinct neck. fig. 1f. on perigona nigriceps (dejean) (coleoptera, carabidae): branice, głubczyce poviat, pile of old hay surrounded by nettles (urtica dioica l.) on the border of a meadow and a cluster of trees (salix fragilis l.) near the opawica river ca. 1 km sww of the village, 15.7.2004, leg. t. m a j e w s k i , tm 10161. laboulbenia manubriolata was described by t h a x t e r (1915) from java and ceylon and reported again from china by r o s s i (1982), who was the first to provide a picture of it. it was later recorded in a number of countries in eastern asia (te r a d a et al. 2004). it seems to occur mostly on perigona nigriceps in asia. this beetle has also been encountered in europe, introduced here sporadically with plant material. its parasite, l. manubriolata, has been reported from portugal (s a n t a m a r i a 1993), great britain (we i r 1996), finland (h u l d é n 1983) and norway (h u l d é n 1985). perigona nigriceps is found in poland very rarely (b u r a k o w s k i et al. 1974 j a ł o s z y ń s k i , s i e n k i e w i c z 2002). some additional laboulbeniales (ascomycetes) new to poland 69 fig. 2. laboulbenia cornuta thaxter. a, b mature thalli (drawn from tm 9420). laboulbenia nana thaxter. c mature thallus (tm 10617), d immature thallus (tm 10618). laboulbe nia olisthopi spegazzini. e immature thallus (tm 10099), f mature thallus (tm 10098). scale bar: 100 μm. 70 t. majewski the features of the only thallus collected in poland are consistent with the descriptions and figures given by, inter alia, r o s s i (1982), s a n t a m a r i a (1993) and te r a d a (2000). laboulbenia nana sugiyama laboulbenia nana sugiyama, ginkgoana 2: 54. 1973. thallus 150-155 μm long, olive-brownish, perithecial region darkened. receptacle 78-108 μm long, curved near the base. cells i and ii 2-3 times longer than broad, cells iii and iv 1.5 times longer than broad, cell v half as long as cell iv. insertion cell dark, slightly constricted; outer appendage long, composed of elongated cells, once branched on second (?) or third cell; basal cell of inner appendage only slightly shorter than basal cell of outer appendage, antheridial branchlets not exceeding the perithecium, but some antheridia may proliferate into longer branchlets. stalk cell of the perithecium oblique, isodiametric or slightly flattened; perithecium ovate, 7074 x 25-30 μm, about 2/3 free, with apical dark spots and prominent round posterior lips (fig. 2: c, d.). on tachyta nana (gyllenhal) (coleoptera, carabidae): rytro, nowy sącz poviat, under the bark of fagus sylvatica l., 28.7.1929, ex coll. szymon te n e n b a u m , tm 10617, 19618 (host in the łomna collection). laboulbenia nana seems to be a very rare species. it was described from japan by s u g i y a m a (1973), later reported only from spain (s a n t a m a r i a 1995, 1998); it may also occur in romania (s a n t a m a r i a 1998). in poland, it has been recorded only on two insects in an old entomological collection. only two mature thalli and two immature thalli growing on the host’s elytron and pronotum were recorded. the thalli are not well preserved, especially appendages; they are, however, consistent with the descriptions and figures given in the above studies by s u g i y a m a and s a n t a m a r i a . the host is a tiny beetle, fairly often encountered under the bark of dead trees in poland (b u r a k o w s k i et al. 1973). laboulbenia olisthopi spegazzini laboulbenia olisthopi spegazzini, redia 10: 55. 1914 (‘l. olistopi’). thallus 190-265 μm long, brownish, perithecial region and upper part of receptacle darkened. receptacle 140-177 μm long, cell i 1.5 times longer than broad in its distal part, cell ii 2 times longer than broad, cells iii and iv nearly isodiametric, cell v half as long as cell iv or slightly longer. insertion cell dark, thick, distinctly constricted; outer appendage long, consisting of elongated cells; basal cell of inner appendage more than half as long as basal cell of the outer one, giving rise to two short antheridial branchlets later proliferating into two strong straight long branches. basal cell of perithecium oblique, rhomboidal, the perithecium 95-130 x 38-45 μm, in 2/3 free, nearly ovate; apex with prominent posterior lips and distinct subapical black spots (fig. 2: e, f.). on olisthopus sturmii (duftschmid) (coleoptera, carabidae): vicinity of przemyśl, leg. tadeusz tr e l l a , no date [before 1940], tm 10099. additional material from ukraine: on o. sturmii, chliwiska, sniatyn poviat, 6.10.1886, leg. stanisław s t o b i e c k i , tm 10098 (both hosts in the kraków collection). the described material comprises 3 immature thalli recovered from the host’s elytron in poland as well as 6 mature thalli and 3 immature thalli growing on a beetle collected in western ukraine. the determination of the thalli is provisional. some additional laboulbeniales (ascomycetes) new to poland 71 laboulbenia olisthopi has been collected across europe (spain, france, germany, italy, switzerland, madeira); only three authors, however, described and illustrated it. the first, s p e g a z z i n i (1914), gives a brief description and somewhat blurred photographs of an immature thallus and three mature thalli, strongly darkened. it seems that cell v is triangular in these thalli, smaller than cell iv and probably is not connected with septum iii/iv. s p e g a z z i n i ’s specimens were collected in italy on olisthopus rotundatus (paykull) and in germany (saxonia) on o. sturmii. b a l a z u c (1975) lists all species of the genus laboulbenia recorded on beetles of the genus olisthopus dejean (odontonyx stephens) up to the time of his study: laboulbenia polyphaga th., laboulbenia olisthopi speg. and laboulbenia flagellata peyr. he identifies olithopus sturmii as the type host of the species described by s p e g a z z i n i , and describes and illustrates the thalli of laboulbenia olisthopi found in france on odontonyx rotundatus (olisthopus rotundatus) and madeira on o. fuscatus dejean. cell v in these specimens is nearly as large as cell iv and is distinctly connected with septum between cells iii and iv. s a n t a m a r i a (1989) describes thalli from spain on olisthopus fuscatus dejean. their cells iv and v are subequal, as in b a l a z u c ’s thalli. similar findings are provided in s a n t a m a r i a ’s successive study (1998): the description and figures show cell v to be as long as cell iv. s a n t a m a r i a concludes that septum iv/v connected with septum iii/iv is an essential character to separate l. olisthopi and forms surrounding l. polyphaga thaxter. undoubtedly, the polish (and ukrainian) thalli collected on olisthopus sturmii and described here belong to a species different from the thalli discussed by b a l a z u c and s a n t a m a r i a , and are likely to be closer to s p e g a z z i n i ’s species. additionally, the host of the present material, olithopus sturmii, is the same host as the one recognised as the type host of l. olisthopi, and the collection site of the type specimen (saxony) is located near poland. „l. olisthopi” thalli visible in the pictures given by b a l a z u c and s a n t a m a r i a come from different hosts and areas situated further away from the present sites. it should be noticed that the thalli determined here as l. olisthopi differ significantly from the thalli of laboulbenia polyphaga recorded in poland (m a j e w s k i 1994) in being clearly darker, and the outer branch of their appendage is not forked. the polish host, olisthopus sturmii, is a xerothermic species, found rarely in the south of poland (b u r a k o w s k i et al. 1974). references b a l a z u c j. 1975. sur les laboulbenia (ascomycètes) parasites d’odontonyx (coleoptera caraboidea, pterostichidae, anchomenini). nouv. rev. ent. 5: 97 100. b a l a z u c j. 1977. deuxième mission biospéologique cubano roumaine à cuba (1973). laboulbéniales (ascomycètes) parasites de coléoptères carabiques. (in:) t. o r g h i d a n et al. (eds). résultats des exspéditions biospéologiques cubano roumaines à cuba, vol. 2, bucureşti: 413 415. b á n h e g y i j. 1944. a balaton környékének laboulbenia féléi. botan. közlem. 41: 49 61, pl. i ii. b u r a k o w s k i b., m r o c z k o w s k i m., s t e f a ń s k a j. 1973. coleoptera, carabidae 1, (in:) katalog fauny polski (catalogus faunae poloniae) 20, pwn, warszawa, pp. 233. b u r a k o w s k i b., m r o c z k o w s k i m., s t e f a ń s k a j. 1974. coleoptera, carabidae 2, (in:) katalog fauny polski (catalogus faunae poloniae) 22, pwn, warszawa, pp. 430. b u r a k o w s k i b., m r o c z k o w s k i m., s t e f a ń s k a j. 1978. coleoptera, histeroidea, staphylinoidea excl. staphylinidae. (in:) katalog fauny polski (catalogus faunae poloniae) 29, pwn, warszawa, pp. 356. 72 t. majewski d e k e s e l a., k r a s t i n a d e k e s e l i. 2006. laboulbeniales (ascomycetes) from latvia. acta my col. 41(1): 55 64. h u l d é n l. 1983. laboulbeniales (ascomycetes) of finland and adjacent parts of the u.s.s.r. karstenia 23: 31 136. h u l d é n l. 1985. floristic notes on palearctic laboulbeniales (ascomycetes). karstenia 25: 1 16. j a ł o s z y ń s k i p., s i e n k i e w i c z p. 2002. the second record of perigona nigriceps (dejean, 1831) (cole optera: carabidae) from poland. wiad. entomol. 20: 173. m a j e w s k i t. 1994. the laboulbeniales of poland. polish bot. stud. 7: 3 466. m a j e w s k i t. 1999. new and rare laboulbeniales (ascomycetes) from the białowieża forest (ne po land). acta mycol. 34: 7 39. m a j e w s k i t., in press. laboulbeniales, (in:) w. w o j e w o d a (ed.), atlas of the geographical distribu tion of fungi in poland. fasc. 4. institute of botany, polish acad. of sciences, kraków. r o s s i w. 1982. new and interesting laboulbeniales from china. mycologia 74: 1023 1026. s a n t a m a r i a s. 1989. el orden laboulbeniales (fungi, ascomycotina) en la peninsula ibérica e islas baleares. ed. espec. soc. catalana micol. 3, barcelona, pp. 396. s a n t a m a r i a s. 1991. el género euphoriomyces (laboulbeniales, ascomycotina). revista iberoamer. micol. 8: 43 50. s a n t a m a r i a s. 1993. contribución al conocimiento de los laboulbeniales (fungi, ascomycotina) ibéricos, iii. orsis 8: 21 31. s a n t a m a r i a s. 1995. new and interesting laboulbeniales (fungi, ascomycotina) from spain, iii. nova hedwigia 61: 65 83. s a n t a m a r i a s. 1998. laboulbeniales, i. laboulbenia. flora mycologica iberica 4, j. cramer, madrid, berlin, stuttgart, 186 pp. s a n t a m a r i a s. 2001. new and interesting laboulbeniales (fungi, ascomycota) from spain, iv. nova hedwigia 72: 375 389. s a n t a m a r i a s. 2003. laboulbeniales, ii. acompsomyces ilyomyces. flora mycologica iberica 5, j. cramer, madrid, berlin, stuttgart, 344 pp. s a n t a m a r i a s., b a l a z u c j., ta v a r e s i.i. 1991. distribution of the european laboulbeniales (fungi, ascomycotina). an annotated list of species. treballs inst. botan. barcelona 14: 1 123. s p e g a z z i n i c. 1914. primo contributi alla conoscenza delle laboulbeniali italiane. redia 10: 21 75, pl. i ix. s u g i y a m a k. 1973. species and genera of the laboulbeniales (ascomycetes) in japan. ginkgoana 2: 1 97, pl. 11 27. te r a d a k. 2000. new records of the carabidicolous laboulbeniales (ascomycetes) of japan (ii). my coscience 41: 39 48. te r a d a k., m. h. h s u , w. j. w u. 2004. notes on the carabidicolous laboulbeniales (ascomycetes) of taiwan i. botan. bull. acad. sinica 45: 87 94. t h a x t e r r. 1895. notes on the laboulbeniaceae, with descriptions of new species. proc. amer. acad. arts sci. 30: 467 481. t h a x t e r r. 1896. contribution towards a monograph of the laboulbeniaceae. mem. amer. acad. arts sci. 12: 187 429, pl. i xxvi. t h a x t e r r. 1915. new indo malayan laboulbeniales. proc. amer. acad. arts sci. 51: 3 51. t h a x t e r r. 1924. contribution towards a monograph of the laboulbeniaceae. part iii. mem. amer. acad. arts sci. 14: 309 426, pl. i xxii. we i r a. 1996. a preliminary host parasite list of british laboulbeniales (fungi, ascomycotina). the entomologist 115: 50 58. nowe dla polski gatunki laboulbeniales (ascomycetes) s t r e s z c z e n i e praca zawiera opisy i ryciny sześciu nowych dla polski gatunków grzybów z rzędu laboul beniales: dimeromyces corynetis thaxter, euphoriomyces magnicellulatus santamaria, laboul benia cornuta thaxter, l. manubriolata thaxter, l. nana sugiyama i l. olisthopi spegazzini. ostatni z nich wykazany został także z ukrainy; dyskutowany jest jego status. 2014-01-01t11:43:32+0100 polish botanical society preliminary study of endophytic fungi in timothy (phleum pratense) in estonia triin varvas, kristina kasekamp and bellis kullman* department of mycology, institute of agricultural and environmental sciences estonian university of life sciences, kreutzwaldi 5, ee-51014 tartu *corresponding author: bellis.kullman@emu.ee varvas t., kasekamp k., kullman b.: preliminary study of endophytic fungi in timothy (phleum pratense) in estonia. acta mycol. 48 (1): 41–49, 2013. timothy (phleum pratense l.) is an important agricultural grass in europe and north america, but there is little research into the occurrence and abundance of fungal endophyte species associated with this grass. the aim of this study was to identify fungal endophytes living within p. pratense and to determine if additional moisture applied during the growing season increases the diversity of endophytic fungi. we studied 58 isolates obtained from surfacesterilised blades of 60 p. pratense plants collected from rõka free air humidity manipulation experimental plots (fahm), estonia. morphological and molecular methods were used for isolate identification. as a result, 45 strains from 10 different taxa were identified, all belonging to ascomycota. five species were found to be new to p. pratense. key words: agricultural grasses, endophytes, isolate identification, its, fahm introduction plants and fungi commonly have a mutualistic relationship and, in the case of endophytes, they reside inside plant tissue, mostly without causing disease (carroll 1988; clay, schardl 2002). sometimes impact from the environment can make these fungi parasitic or pathogenic to host plants (schardl et al. 1997; meijer, leuchtmann 1999; schulz et al. 1999; kogel et al. 2006; johnson, oelmüller 2009). most of them are non-specific regarding their host preference (petrini 1986; saikkonen et al. 1998; arnold et al. 2000) and can live together with other endophytes in the same host (an et al. 1992; saikkonen et al. 1998; arnold et al. 2000), holding greater promise for plant growth promotion and increased yield in agriculturally important grasses. endophyte presence in plants has also been shown to help them deal with prolonged periods of drought, excess moisture or light (bouton et al. 1993; bier 1995; marks, clay 1996; lewis et al. 1997; cheplick et al. 2000; malinowski, belesky 2000). acta mycologica vol. 48 (1): 41–49 2013 doi: 10.5586/am.2013.006 42 t. varvas et al. increases in biomass and tolerance to attack from herbivores and insects are often ascribed to endophytes (bacon, hill 1996; clay and holah 1999; sahay, varma 1999; redman et al. 2001; arnold et al. 2003; waller et al. 2005; márquez et al. 2007). p. pratense is a widespread arable crop that is, however, poorly investigated in terms of its endophytic fungal diversity. the purpose of our study was to identify fungal endophytes living inside the leaves of p. pratense and to determine if differences in water treatments in the fahm experimental area affect the diversity of endophytes. materials and methods sampling plant material and endophyte isolation. samples were collected on 29 october 2008 from free air humidity manipulation (fahm) experimental plots at rõka, tartu county, in southeast estonia (58°14’44”n, 27°17’58”e). plants were collected from four plots with additional moisture being applied to the plots (h1-h4) and from two control plots without additional water being applied (c1 and c4). in one plot, h3, there appeared a problem with the watering system and it could not be used in comparison. altogether, 60 p. pratense plants were collected. immediately after collection, one to two p. pratense leaves were separated from the plants and cut into 5 × 5 mm fragments. the surfaces of the fragments were disinfected by rinsing in 96% ethanol for 1 min, followed by repeated rinsing in sterile water. fragments were placed in petri dishes (9 cm diam.) on potato dextrose agar (pda, merck kgaa germany) supplemented with tetracycline and streptomycin (50 μl each per ml) for suppression of bacterial growth. petri dishes were examined for mycelial growth for 30 days and all fungal growth was subcultured on individual pda plates with antibiotics for subsequent identification. for molecular identification, 7-10 days after inoculation of subcultures, a 5 × 5 mm piece of mycelium with agar was cut and preserved in sodium dodecyl sulphate (sds). endophyte identification by morphology. morphological characterization relied on the colour, appearance, growth rate and microscopical hyphal features of endophytic colonies. similar cultures were grouped together based on morphotype and the main characteristics were analysed for representatives of these groups. growth rate was measured for 23 days. image pro plus 6.0 program was used to calculate the area occupied by mycelium. colony colour was determined at approximately 17 days using the methuen handbook of colour (kornerup, wanscher 1967). both surface and reverse colours were recorded and in case of more than one dominant colour, all were recorded (data not shown). colony colour was determined by viewing the isolates in daylight. the colour of the medium and the occurrence of aerial mycelium were also noted if present. morphological descriptions were made in sterile water using a zeiss axioskop 40 fl microscope, an axiocam mrc camera and the axio vison 1.6 program. species determination followed taxonomic monographs and manuals (domsch et al. 1980; ellis, ellis 1997; leslie, summerell 2006). vouchers strains were deposited in the collection of fungal living cultures [tfc 2010-001 – tfc 2010-035] at the estonian university of life sciences. endophytic fungi in timothy in estonia 43 endophyte identification by molecular methods. genomic dna was extracted from pure cultures using genomic dna purification kit k0512 (fermentas) and stored in ctab buffer according to gardes and bruns (1993). its regions of rdna were amplified by pcr using primers its1f (5´ to 3´: ctt ggt cat tta gag gaa gta a) and its4 (5´ to 3´: tcc tcc gct tat tga tat gc) using a biometra tprofessional basic thermocycler (biometra gmbh, germany) with 35 cycles of 30 s at 95°c (denaturing), 30 s at 55°c (annealing), and 1 min at 72°c (extension). the resulting pcr product was purified with exo-sap (fermentas) following manufacturer’s instructions. samples were sequenced in macrogen (korea) with primers its1 5` to 3`: tcc gta ggt gaa cct gcg g and its5 5` to 3`: gga agt aaa agt cgt aac aag g. sequences were edited using the program sequence scanner 1.0 (applied biosystems, foster city, california, usa). sequences obtained from 45 cultures were compared with the ones in genbank using blastn algorithm. sequences that were more than 97% identical were considered to be homospecific. colonization frequency (cf%, the percentage of plants colonized) was calculated by using the formula: cf = (ncol/nt) × 100, where, ncol is the number of segments colonized by each endophyte and nt is the total number of segments observed (hata, futai 1995). results in total, 58 endophyte strains were isolated from the 60 plants. of these, 45 isolates were identified by sequence analysis of the its region. these isolates belonged to 10 taxa based on sequence data its: alternaria arbusti e.g. simmons, lewia viburni e.g. simmons (anamorph alternaria viburni e.g. simmons), apiospora montagnei sacc. (anamorph arthrinium arundis (corda) dyko et b. sutton), aureobasidium pullulans (de bary) g. arnaud, epicoccum nigrum link, fusarium sp., gibberella avenacea r.j. cooke (anamorph fusarium avenaceum (fr.) sacc.), monographella sp., paraphaeosphaeria michotii (westend.) o.e. erikss. and phaeosphaeria herpotri­ choides (de not.) l. holm (fig. 1.). species determinations using genbank references are given in table 1. colonization frequency of isolated fungi is shown on figure 2. morphological studies were performed for most isolates, but only 33% of them resulted in determination of species or genus, while others remained sterile. morphological studies allowed identification of six species or genera; a. pullulans, e. ni­ grum, f. sporotrichioides, f. avenaceum,, arthrinium sp. and alternaria sp. (fig. 1). a plant sample usually contained only one fungal species, although two of the samples gave two different species each (e. nigrum and a. arbusti, e. nigrum and p. herpotri­ chioides). in morphological analyses, 67% of cultures were sterile and for these its analysis and morphotype concept (guo et al. 2000; arnold et al. 2003) was used to determine species. growth rates had noticeable variation both between species (0.08–0.71 cm/day) and also within species. growth rate within the e. nigrum group was 0.10–0.26 cm/ day. average growth rate for all species combined was 0.25 cm per day, with a. pul­ lulans having the lowest (0.08) and a. montagne the highest (0.71) radial growth. 44 t. varvas et al. the results of our study showed that the diversity of endophytes was not higher in the experimental plots with additional water given to the plants during the growing season. in fact, both the highest number of isolates (plot h4, 15 isolates) and the lowest number of isolates (plot h1, 5) were obtained from plots with applied additional moisture. in average, 11 isolates were obtained from each control plot and 9 isolates were obtained from each experimental plot. considering these results, the positive effect of extra moisture on the diversity of endophytic fungi was not established. table 1 endophytic fungi isolated based on sequence comparison to genbank database isolate species genbank accession no. h4-8-2 phaeosphaeria herpotrichoides jn688915 h3-8-3 apiospora montagnei jn688916 c1-7 aureobasidium pullulans jn688917 c1-6 epicoccum nigrum jn688918 c1-4-3 alternaria arbusti jn688919 h3-6-1 fusarium avenaceum jn688920 h4-4-1 alternaria viburni jn688921 c1-9-1 fusarium sporotrichioides jn688922 c4-7 monographella sp. jn688923 c1-3-3 paraphaeosphaeria michotii jn688924 fig. 2. identified species and their colonization frequency (cf%). endophytic fungi in timothy in estonia 45 fig. 1. photographs of fungal cultures (surface and reverse): a – alternaria arbusti, b – phaeo­ sphaeria herpotrichoides, c – gibberella avenacea (fusarium avenaceum, d – fusarium sporo­ trichioides, e – apiospora montagnei (arthrinium arundis), f – paraphaeosphaeria michotii, g – aureobasidium pullulans, h – monographella sp., i – epicoccum nigrum, j – epicoccum nigrum, lewia viburni (alternaria viburni) photograph not included. 46 t. varvas et al. discussion this study of the endophytes of p. pratense, a temperate perennial grass, resulted in the identification of 45 strains from 10 different taxa. according to previous research, the presence of endophytes can be higher in tropical plants (arnold, lutzoni 2007), approaching 100%. in plants from colder climates the endophyte diversity is much lower (rosa et al. 2009). the rate of endophytes is also dependent on environmental conditions. results from zamora et al. (2008) suggest that the season, isolation method and tissue health decisively affected the frequency and species distribution of endophytes. almost all species isolated in zamora et al. (2008) work were recovered in autumn and nearly half of these exclusively during this season. the fungal endophytes associated with p. pratense have previously been investigated mostly within large scale studies along with other species and in relation to their toxicity to animals, differences in infection rates in natural and cultural communities and diversity (bacon, hill 1996; clay, holah 1999; sahay, varma 1999; redman et al. 2001; arnold et al. 2003; waller et al. 2005; márquez et al. 2007). earlier studies have found, from p. pratense, mainly ascomycota, but also basidiomycota, blastocladiomycota, chytridiomycota and zygomycota (waipara et al. 1996; ellis, ellis 1997; sanchez marquez et al. 2010). these species range from saprobes to pathogens, but some might turn out to be endophytes as well. it is sometimes difficult to differentiate between pathogens and parasites or epiphytes and endophytes. transitions may occur between these types (redlin, carris 1996; azevado 1998). most research concerning p. pratense has so far dealt mostly with pathogens, epiphytes and saprobes. for a complete list of previously described fungi connected with p. pratense see the following resources: newcrop (http://www.hort. purdue.edu/newcrop/duke_energy/phleum_pratense.html), ellis and ellis (1997), ing (1990). some of these species are known to have only one type of association with the host plant, while others have been found to be either epiphytic, endophytic, saprotrophic or pathogenic, depending on the host plant. this suggests that the type of association is not always constant and depends on many variables. all of the 10 endophytes identified in this study belong to ascomycota, four of which were anamorphs for whom teleomorphs connections are unknown or not recorded according to index fungorum (www.indexfungorum.org). these are a. ar­ busti, a. pullulans, e. nigrum and f. sporotrichioides. p michotii, p. herpotrichoides, and mostly also monographella sp. teleomorphs connections with anamorphs are problematic and mostly unknown or not recorded. only three of our species have known connections between teleand anamorphs: a. montagnei is associated with a. arundinis, g. avenacea with f. avenaceum and l. viburni with a. viburni. in terms of abundance, we found in our study, that e. nigrum was the dominating species (colonizing frequency 67% of all isolates (fig. 2). when one plant sample was shown to have more than one endophyte, e. nigrum was always present. sanchez marquez et al. (2010) also referred to the high domination rate of holcus lanatus l. by epicoccum, alternaria and aureobasidium. in terms of fungal diversity based on different water treatments, no definite positive effect of additional water availability was observed. endophytic fungi in timothy in estonia 47 conclusions the aim of this study was to identify fungal endophytes living inside the leaves of p. pratense. as a result, 45 strains from 10 different taxa were identified. half of the taxa isolated in this study – a. pullulans, e. nigrum, g. avenacea, monographella sp., and p. herpotrichoides – had been identified from timothy earlier as endophytes (salkin et al. 1986; domch et al. 1993; huang et al. 2008; osono 2008; tao et al. 2008; dodd 2010;). a. arbusti, l. viburni, a. montagnei, f. sporotrichioides and p. michotii are new endophytic fungi isolated from timothy. two of these, a. arbusti and l. viburni, are also new species to estonia (varvas, kullman 2012). to obtain a better insight into the communities of blade endophytes, in situ molecular techniques should be used for unculturable and slowly growing fungal taxa. in particular, 454 sequencing allows to identify of hundreds of species using a large number of samples (jumpponen, jones 2009). acknowledgements. the research was supported from the project “plant protection for sustainable crop production” sf0170057s09. we thank dr. leho tedersoo (institute of ecology and earth sciences, university of tartu) for useful consultations and help. our sincere thanks are due to mrs. e. jaigma for revising the english text of the manuscript. references an z.q., liu j.s., siegel m.r., bunge g., schardl c.l. 1992. diversity and origins of endophytic fungal symbionts of the north american grass festuca arizonica. theor. appl. genet. 85: 366-371. arnold a.e., maynard z., gilbert g.s., coley p.d., kursar t.a. 2000. are tropical endophytes hyperdiverse? ecol. let. 3: 267-274. arnold a.e., mejia l.c., kyllo d., rojas e., maynard z., robbins n., herre e.a. 2003. fungal endophytes limit pathogen damage in a tropical tree. proc. natl. acad. sci. usa 100: 15649-15654. arnold a.e., lutzoni f. 2007. diversity and host range of foliar fungal endophytes: are tropical leaves biodiversity hotspots? ecology 88: 541-549. azevedo j.l. 1998. microorganismos endofíticos. ecologia microbiana, embrapa, jaguariuna. bacon c.w., hill n.s. 1996. symptomless grass endophytes: products of coevolutionary symbioses and their role in the ecological adaptations of grasses. 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university of wrocław przybyszewskiego 66/73, pl-51-148 wrocław, jurekp@microb.uni.wroc.pl piątkowski j., świątek m.: effect of aminoacids on the fungicidal activity of quaternary ammonium salts. acta mycol. 45 (2): 207–217, 2010. amphipatic compounds exhibit an antimicrobial action both on bacteria and fungi. it is caused by a penetrative property of hydrophobic carbon chain of the compuound into a plasma membrane as well as by additional interaction of membrane elements and a hydrophilic amphipathic compound head. bactericidal and fungicidal activity of this compound strongly depends on chemical environmental factors. in general, microorganisms are not as sensitive in a full medium as in a minimal one and the level of sensitivity rises when the amphipatic compounds are presend in destilled water. similarly, the sensitivity is stronger in fluid than on solid medium. our researches revealed however that some aminoacids, although they are complex organic compounds, increase the microbial sensitivity to some tested compound. this efect depends on a microorganism and on a kind of compound. the highest hipersensitivity has been observed against yeast-like fungi when arginine was a cooperating aminoacid. the effect concerns trichosporon but not e.coli, not occurs in relation to sds, quaternary ammonium salt ia, and bisammonium salts. certainly the effect exhibit qas, which have simple composition of hydrophilic „head” consisting only of methyl group, attaching to alkilic chain possessing keton group, build of 14 or 16 carbon atoms. key words: amphipathic compound, quaternary ammonium salts, yeast, trichosporon introduction quaternary ammonium salts (qas) are ammonium ion nh4 + derivatives, in which all the four hydrogen atoms have been replaced with organic substituents. as the nitrogen atoms are negative, the ammonium group is hydrophilic. if one of the substituents is a hydrophobic alkyl chain, it confers the molecule an amphiphilic character. qas are found in living organisms where they perform physiological functions. trimethyllysine, which is involved in the mitogenesis of human limphocites acta mycologica vol. 45 (2): 207–217 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 208 j. piątkowski and m. świątek (suba et al. 1980), has been detected in proteins, confering them specific features (houtz et al. 1989). carnitine enables transport of fatty acids through the inner mitochondrial membrane (stryer 1999). betaino-glycine is utilized by bacteria, algae, higher plants and animals for osmoregulation (weretilnyk et al. 1989). homarine acts in crustacea as a donor of methyl group (netheron, gurin 1982). there are also qas that have long been known for their toxic activity. charamine, a derivative of aliphatic amino acids, is an antibiotic produced by some algae. fascaplysin, a derivative of indol, is a red pigment toxic to microorganisms (roll et al. 1988), obtained from a marine sponge. the heterocyclic compound bromogramine inhibits mitosis (sato, fenical 1983). the basic mechanism responsible for the toxic effect of amphiphilic compounds depends on their affinity for cell membranes. at a high qas concentration, pores filled with water may occur in the plasma membrane. the walls of the pores are composed of hydrophilic heads of qas (kuczera 1983). the salts may also bind to a specific site of the plasma membrane protein, thus changing its conformation (helenius, simons 1975). in this way, the activity of the plasma membrane enzymes has been inhibited (samuel 1972; baltscheffsky h., baltscheffsky m. 1974). enzymatic activity will also be inhibited when a lipid supporting the activity of the enzyme has been washed away by the qas (coleman, allison 1973). the mechanism governing the action of qas on fungi depends on the composition of the compounds. it has been found that qas act at the plasma membrane level (kołodyński et al. 1984; skała et al. 1988; piątkowski et al. 1990; lachowicz, piątkowski 1995) and at the level of intracellular structures – mitochondria or lisosomes (hussain et al. 1987). an important factor influencing the yeast cell sensitivity to qas is the cell genotype (obłąk et al. 1988; piątkowski et al. 1990; lachowicz et al.1992). in the study reported on in this paper, the effect of enhanced qas toxicity against yeast and trichosporon cutaneum in the presence of some aminoacids was examined. especially arginine was found to be powerful in enhancing the effect of tetradecacarbonylmethyloxy-n,n-trimethylammnium chloride on the fungi. this effect was also observed with a similarly constructed qas, which differed only in the length of the alkyl chain. materials and methods the qas being studied were obtained from the department of chemistry, wrocław university of technology, by prof. stanisław witek. the prototrophic yeast strain saccharomyces cerevisiae sm5 came from the institute of microbiology, university of wrocław; trichosporon cutaneum ucd 54169 from the institute of biophysics and biochemistry polish academy of sciences, warsaw; escherichia coli pcm 2057, pseudomonas aeruginosa pcm 2058 and staphylococcus aureus pcm 2054 from the institute of immunology and experimental therapy polish academy of sciences, wrocław. effect of aminoacids 209 all the strains were stored at minus 70°c in the appropriate glycerol medium. they were precultivated for 24 hours in liquid full medium: yeast and trichosporon in ypg (glucose 2%, yeast extract 1%, bacto peptone 1%), bacteria in nutrient broth. after translocation of 10 μl of the preculture to 10 ml of such medium, the microorganisms were incubated until the logarithmic phase of growth was attained. in order to estimate the qas concentrations at which 100% of cells is killed or their growth inhibited, the cells were washed twice by centrifugation in physiological liquid (0.9% nacl) and then placed with an inoculating loop on the medium suplemented with the qas being tested. the compounds concentrations were: 5, 10, 20, 40, 80, 160, 320, and 640 μm. the medium used for yeast–like organisms was ypg or minimal synthetic medium ynb; for bacteria – nutrient broth or minimal davies medium (k2hpo4, 0.7%; kh2po4, 0.2%; sodium citrate, 0.05%; (nh4)2so4, 0.1%; mgso4 x 7h2o, 0.01%; glucose, 1%). ynb medium and davies medium were supplemented, if necessary, with 3% of casamino acids or aminoacids at a concentration of 40 μg/ml. after 24 hours of fungusor 12 hours of bacteria incubation the growth of these microorganisms was detected to assign the compound concentration at which the growth of 100% of the cells were inhibited. in the case of the liquid media, the cells after a compound treatment were spreaded onto full agar medium to verify their viability. before the cells were planted onto the media with the inoculated loop, they were dispersed in physiological liquid to the optical density of 0.05 measured by table 1 compounds used in investigations compound chemical formula n-dodecyl-n, n-dimethyl-n-[3-(β-methylo-βnitrovinyl)6-methoxybenzyl] ammonium chloride ( ia ) * mw 466 och3[cl –] ch2 —n +(ch3)2 no2(ch3)c=ch c12h25 hexadecacarbonylmethyloxy-n, n-trimethylammonium chloride * mw 377.5 c16h33 —coch2o — n +(ch3)3 [cl –] tetradecacarbonylmethoxy-n, n-trimethylammonium chloride * mw 349.5 c14h29 —coch2o — n +(ch3)3 [cl –] decacarbonylomethyloxy-n, n-trimethylammonium chloride * mw 293.5 c10h21 — coch2o — n +(ch3)3 [cl –] sodium dodecyl sulphate (sds) * mw 288.4 h3c — (ch2)10 — ch2oso3 – [na+] decacarbonylmethyloxy-n, n-trimethylbisammonium chloride (c10bis) ** mw 585.0 c10h21 — coch2o — n +(ch2)2 [2cl –] ch2 ch2 c10h21 — coch2o — n +(ch2)2 explanations: * monoammonium salts, ** bisammonium salts, mw – molecular weight. 210 j. piątkowski and m. świątek spectrophotometry at 550 nm. the solid media on which the microorganisms were cultivated had the form of a slant in a test-tube. the activity of the compounds in the liquid medium was verified by pouring the cells of trichosporon into 10 ml of ynb and ynb with amino acids, at the concentration of 104 cells/ml. after 30 min. of incubation at 28°c, 10 μl samples of the cell suspension were collected and placed on the full solid medium in order to estimate viability. since the distance between the compound concentration used was long (the next one was twice as high as the former one), the results obtained were in high degree repeated. that’s why no variation values has been marked in the figures. results in order to verify to what extent the sensitivity of trichosporon cutaneum depends on the medium composition, the cells were placed in three types of the growth medium (described in the section „materials and methods”): ypg, ynb with glucose as the only carbon source, and ynb with glucose and casamino acids as the source of amino acids. in each type of medium different concentrations of qas were used. the viability of the cells was evaluated after incubation for 3 days at 28°c. the results are gathered in figure 1. as shown by this data, the cells on ypg and ynb + casamino acids were viable when the concentration of the c14 compound was lower than 40 μm. on the other hand, growth on the medium without amino acids was visible even at the c14 concentration of 160 μm. this implies that the fig. 1. growth of t.cutaneum on the medium ypg,ynb and ynb+casamino acids at the presence of c14 salt. effect of aminoacids 211 sensitivity of the strain t. cutaneum to c14 in the absence of amino acids is approximately four times as high as when they are present in the medium. taking into account the considerable difference in sensitivity, we decided that the method is sensitive enough to futher investigations. upon analysis of the results, a number of detailed guestions arise: is the effect of supersensitivity in the presence of aminoacids characteristic for all amphiphilic compounds, or only for a narrow group of molecules similar to one another? it is only the c14 salt that produces such an effect? to answer thise questions, some other compounds were tested for their activity against t. cutaneum. two of them differed from c14 only in that they had a c10 and c16 carbon atoms in alkil chain respectively. the third and fourth was bisammonium compounds: one – c10, the other – c16. the fifth salt, so colled ia, differed in the hydrophilic „head” composition. the last one of the compounds tested was sds, which is not an ammonium salt and, in contrast to the other mentioned, occurs in water as an anion. the results of the tests with all the compounds mentioned are presented in figure 2. as shown by the results, (fig. 2) apart from c14 this is the c16 monoammonium salt alone that produces the effect of amin oacid-dependent hipersensitivity, although the effect is poorer than the one developed by c14. possesing 10 carbon atoms c10 qas was equally active both in the ynb and the ynb+a medium, where the growth of t. cutaneum became visible when the compound concentration did not exeed 320 μm. hipersensitivity was not observed with ia qas and sds. a question was whether or not the influence of amino acids on the sensitivity of the microorganisms was specyfic for t. cutaneum? another yeast-like organism, saccharomyces cerevisiae, was chosen to carry out tests that would help to answer this fig. 2. growth of t.cutaneum on ynb and ynb+ca at the presence of amphifilic compounds. 212 j. piątkowski and m. świątek question. two species of bacteria were also investigated for sensitivity – echerichia coli and pseudomonas aeruginosa. as it can be infered from the results in figure 3, hypersensivity is observed with s. cerevisiae but not with the bacteria used. if the effect is stimulated by the medium casamino acids, it is obviously interesting to detect what kind of amino acids increases the sensitivity to those qas. in order to verify this, the next experiment did not involve casamino acid medium; use was made of particular amino acids present in the ynb medium. one different amino acid at concentration in the medium 40 μg/ml was present in each variant of the media. as shown by the data in the tabele 2 shows, each of the amino acids being studied affected the sensitivity of t. cutaneum to a different extent. the greatest growth inhibition was observed, when ynb was supplemented with arginine. to a slighty smaller degree than by arginine was the antifungal activity of the qas enhanced by uracyl, tryptophan, cysteine and leucine. significantly lower inhibition occured from thyrosine, lysine. only a slight inhibiting efect was produced by treonine and glutamic acid. no increase in sensitivity was found in the presence of adenine, histidine or proline. the experiments involving solid media did not provide information as to whether the qas had a toxic or only an inhibitory effect on fungal growth. if this had been an inhibitory effect without cell destruction, restoration of growth should have been detected when the cells were separated from the c14 (after previous treatment with this salt), and brought into a fresh medium free from c14. we used a liquid medium where trichosporon cells were incubated in the presence of c14 and thereafter fig. 3. effect of supersensitivity of e.coli, p.aeruginosa, s.cerevisiae and to c14 compound. effect of aminoacids 213 placed on the ypg solid medium which was free from c14, in order to assess how many cells had survived. the results of the experiment indicate that when the cells were incubated with c14 in the ynb liquid medium, the critical compound concentration which killed them amounted to 5 μm. when they were grown in ynb with amino acids, the killing concentration was 0.55 μm (fig. 4). table 2 growth of t. cutaneum on ynb suplemented with c14 and an aminoacid aminoacid concentration of the c14 compound [μg/ml] 0 20 40 80 160 320 leu + + + − − − pro + + + + + + his + + + + + + lys + + + + − − arg + + − − − − cys + + + − − − glut + + + + + − met + + + − − − ala + + + − − − thr + + + + + − ade + + + + + + tyr + + + + − − trp + + + − − − ura + + + − − − glyc + + + − − − fig. 4. viability of trichosporon cutaneum cells in the presence of c14 compound and aminoacids. ynb – a medium without aminoacids; ynb+ca – a medium with aminoacids 214 j. piątkowski and m. świątek the sensitivity of trichosporon was tested using two media: ynb and ynb supplemented with amino acids in form of casamino acids (ynb+ca). figure 5 and 6 show the results of the test, in which the cells were inoculated on ynb and ynb+ca slants in such a number that made the growing colony a countable one. as shown by the photographs, the growth of the cells was more sensitive when amino acids were present in the medium. it is interesting to note that when the c14 concentration was lower than the inhibiting one, colony growth was enhanced to a greater extend in the presence than in the absence of casamino acids for the same qas concentrations. however, with c14 concentration of 20 μm and higher, casamino acid stopped supporting microbial growth and started inhibitig it. discusion quaternary ammonium compounds are widely used as remedies for killing microorganisms. their inhibitory activity against bacteria (jaszczuk, krzywicka 1975; zabłocki et al. 1970), fungi (witek et al. 1978), algae (rucka et al. 1983) and, also, agaist insects (fulde 1976) have long be known. desinfectant preparations applied in hospitals are available on the market (kędzia 1981). however, qas are not normaly used as drugs for direct contact with a human body. in such cases, a variety of antibiotics has been applied, but their mechanism of action is not based on amphiphilic properties. the results of the research reported on in this paper shows clearly that the action of a quaternary monoammonium salt with 14 carbon atoms in the alkyl chain on trichosporon cutaneum can be increased approximately fourfold if the environment includes amino acids. our tests with the liquid medium substantiate not only an inhibitory but also a killing mechanism that governs the action of the compound. the toxic effect is much stronger in a liquid than on a solid medium. the increase in sensitivity due to the interaction with amino acids does not occur in all organisms. there in no doubt that it would be interesting to carry out such tests with a greater number of different living cells. the problem is important and worth studying, so will be subject to further experiments reported in a separate paper in the future. the purpose of our study reported on in this paper was to provide the widest possible background to a comprehensive analysis of the effect of „cooperation” between amino acids and qas. the effect occurs in trichosporon but not e. coli or probably other gram negative rods of bacteria. effect of supersensitivity induced by the presence of amino acids not occurs does not occur when the trichosporon strain is treated with sds, quaternary ammonium salt ia or bisammonium salts. at the present stage of our investigation it is difficult to decide which element in the structure of the compounds is essential in this context. the effect is certainly associated with those qas that have simple composition of hydrophilic „head” where there is only a methyl group attached to alkylic chain possesing a keton group build of 14 or 16 carbon atoms. it is still unknown whether or not an alkilic chain without the keton group would develop such an effect. further tests are needed to define the exact structural requirements triggering this effect. 40 μm 80 μm 160 μm 320 μm fig. 5. growth of trichosporon cutaneum on the medium ynb at the presence of c14 compound. 5 μm 10 μm 20 μm 40 μm fig. 6. growth of trichosporon cutaneum on the medium ynb at the presence of c14 compound and amino acids. effect of aminoacids 215 the results obtained so far indicate that the stimulation of the fungicidal activity of amfphifilic compounds by amino acids is not a common feature of the qas, it probably pertains to only a narrow group of them. the research on particular amino acids revealed an important role of arginine in increasing the sensitivity of yest-like organisms to c14. as far as the nutrients being tested are concerned, this rise in sensitivity does not seem to be caused by compounds having a cyclic composition, e.g., proline, histydyne or adenine. on the basis of the results obtained up till now it is difficult to explain the mechanism of cooperation between aminoacids and c14. what should further investigations concern on? maybe it is necessary to verify whether or not the interaction of amino acids with the amino acid transport system through the plasma membrane activates the carriers transporting c14 towards a cell? mayby the association of arginine to permease changes the conformation of the protein in such a way that it becames more sensitive to inhibition by c14. it is well known that amphiphilic compounds which act on proteins at appropriately low concentrations may specifically bind to some sites of this protein. if the protein changes its conformation, the enzymatic activity may become inhibited. the chemical composition of both arginine and c14suggest that they are likely to fuse and that arginine transported by permease into the cells might enable co-transport of c14. summing up, with the results obtained it is possible to characterise the interesting phenomenon of amino acids induced increase in the fungicidal activity of the qas. this effect is interesting for cognitive reasons, as it may contribute to a better understanding of the mechanism governing the action of quaternary ammonium compounds on microorganisms. of equal importance seem to be the therapeutic aspect of this phenomenon. our results indicate that the observed supersensitivity is detected in yeast-like organisms, not in gram negative bacteria. if further researches revealed that also epithelial cells are resistant to this effect, this would means that the aminoacid – c14 combination is a remedy against trichosporon, which is believed to be a pathogenic microorganism colonising espetially throat. on the other hand, some gram negative bacteria, such as moraksella catharalis, play an important role in upper respiratory tracts. so it should be advantageous to have a drug which kills microorganisms such as trichosporon and leaves the natural and useful bacteria intact. a promising result is that c14 acts significantly stronger in a liquid environment. the presented results disclosed some basic features of the observed phenomenon of the amino acid induced rise in the activity of qas. it is obvious that the final explanation of the mechanism of cooperation between the compounds needs further experiments. 216 j. piątkowski and m. świątek references baltscheffsky h., baltscheffsky m. 1974. electron transport phosphorylation. annu. rev. biochem. 43: 871–897. coleman r., allison a. c. 1973. membrane-bound enzymes and membrane ultrastructure. biochim. biophys. acta 300: 1–30. fulde s. 1976. rozwój badań w zakresie syntezy i przygotowania produkcji enolofosforanów owadobójczych. pestycydy 4: 1. helenius a., simons k. 1975. solubilization of membranes by detergents. biochim. biophys. acta 415: 29–71. houtz l., stults j. t., mulligan r. m., tolbert n. e. 1989. post-translational modifications in the large subunit of ribulose biphosphate carboxylase/oxygenase. proc. natl.acad. sci. u.s.a. 86: 1855–1859. hussain m., leibowitz m. j., lenard j. 1987. killing of saccharomyces cerevisiae by lysosomatropic detergent n-dodecylimidazole. antimicrobial agent and chemotherapy 512–517. jaszczuk e., krzywicka h. 1973. oporność bakterii na działanie niektórych kationowych związków powierzchniowo czynnych. roczn. pzh 24: 241. kędzia w. b. 1981. dezynfekcja w medycynie i farmacji. pwn, warszawa. kołodyński j., ułaszewski s., grobelny d., witkowska r., witek s., lachowicz t. 1984. effect of some quaternary benzylammonium salts on physiology of yeast. acta microbiologica polonica 33: 119– 130. kuczera j. 1983. wpływ amfifilowych detergentów na błony liposomów. zagadnienia biofizyki współczesnej. pwn warszawa 53–101. lachowicz t. m., piątkowski j. 1995. quaternary ammonium salts and arginine are inhibitors of general amino acid permease in yeast. pestic sci. 43: 169–180. lachowicz t.m., obłąk e., piątkowski j. 1992. auksotrophy stimulated sensitivity to quaternary ammonium salts and its relation to active transport in yeast. bulletin of polish academy of sciences. biological scien. 40: 173–182. netherton j. c., gurin s. 1982. biosynthesis and physiological role of homarine in marine shrimp. j. biol. chem. 257: 971–1011. obłąk e., ułaszewski s., lachowicz t. m. 1988. mutants of saccharomyces cerevisiae resistant to a quaternary ammonium salt. acta microbiologica polonica 37: 261–269. piątkowski j., orłowska g., lachowicz t. m. 1990. sensitivity of aminoacid transport in yeast to amonium salts. (in:) genetic of respiratory enzymes in yeasts. wrocław university press. 103–113. roll d. m., ireland c. m., lu h. s. m., clardy j. 1988. fascaplysin, an unusual pigment from the marine sponge fascaplysinopsis sp. j. org. chem. 53: 3276–3278. rucka m., oświęcimska m., witek s. 1983. new biocides for cooling water treatment quaternary ammonium salts derivatives of glycine esters. environ. protect. enqin. 9: 25–31. sato a., fenical w. 1983. gramine-derived bromoalkaloids from the marine bryozoans zoobotryon verticillatum. tetrahedr. lett. 24: 481–484. samuel r. 1972. reconstitution of biological membranes. biochim. biophys. acta 265: 241–296. skała j., orłowska-matuszewska g., misiewicz m., oświęcimska m., witek s., kotylak z. 1988. biological effect of alkoxymethylene trimethylammonium chlorides on yeast saccharomyces cerevisiae. acta microbiologica polonica 37: 271–280. stryer l. 1999. biochemia. pwn warszawa. suba z., szende b., lapis k., takacs j., elek g. 1980. ε-n-trimethyllysine, a natural cell component with mitogenic activity. neoplasma 27: 11–23. weretilnyk e. a., bednarek s., mccue k. f., rhodes d., hanson a. d. 1989. comparative bichemical and immunological studies of the glycine betaine synthesis pathway in diverse families of dictyledons. planta 178: 342–352. witek s., grobelny d., ptaszkowska j., bielecki a., bakuniak e., fulde s., górska-poczopko j. 1978. belg.patent no 864782. zabłocki b., kotełko k., szydłowski s., gromska w., izdebska k., czerniawski e., michno-bednarek z., gościcka t., sedlaczek l. 1970. dokumentacja polfy, sterinol. 5:5. effect of aminoacids 217 wpływ aminokwasów na grzybobójcze działanie czwartorzędowych soli amoniowych streszczenie jednoamoniowa sól czwatorzędowa z 14 atomami węgla w łańcuchu węglowym wykazuje silniejsze działanie letalne wobec trichosporon cutaneum na podłożu pełnym oraz podłożu opartym na hydrolizacie kazeiny niż na podłożu minimalnym. taki efekt uzyskuje się również z solą o 16 węglach w łańcuchu. w przypadku dwuamoniowych związków tego typu, sds oraz soli amoniowej z aromatycznym pierścieniem, wzrostu wrażliwości w obecności aminokwasów nie zaobserwowano. poza t. cutaneum większa wrażliwość w obecności aminokwasów wystąpiła również w przypadku drożdży, lecz nie w przypadku testowanych bakterii pseudomonas sp. i escherichia coli. największą aktywność grzybobójczą soli amoniowych uzyskano dla argininy. efekt ten jest znacznie silniejszy w podłożu płynnym niż stałym. 2014-01-01t11:51:29+0100 polish botanical society in memoriam professor dr alina skirgiełło (1911-2007) professor alina skirgiełło, the founder and editor of acta mycologica, passed away on 10 october 2007. her death has dealt a severe blow not only to polish mycologists, but also to many of her friends and colleagues worldwide. born in klince, ukraine, on 3 november 1911, skirgiełło attended schools in białowieża and grodno as her family followed her forester father. in 1931, she entered warsaw university, where she studied natural sciences, graduating six years later after the completion of the diploma dissertation in mycology polish terrestrial tube fungi supervised by professor bolesław hryniewiecki at the department of systematics and plant geography. skirgiełło soon joined the department, first as a scholarship holder of governmental foundations and later as a lecturer (1 january 1939). the war, however, forced her to work as a gardener. a member of the polish home army, she was active in the resistance movement. it was largely through her efforts that the valuable book collection and herbaria of the department were saved from annihilation by the nazis. in 1945 skirgiełło returned to the department of systematics and plant geography, warsaw university, as a senior lecturer, and remained affiliated with it until her retirement in 1982. the thesis the genus russula in poland earned her a doctoral degree in 1948. she was appointed reader in 1954, extraordinary professor in 1964 and full professor in 1972. she was head of the department from 1960 until 1979, served as pro-dean and dean of the faculty of biology, warsaw university, between 1963 and 1975, and director of the institute of botany, warsaw university, between 1975 and 1979. while actively engaged in research and teaching at warsaw university, skirgiełło was also deeply committed to other causes. she belonged to the polish botanical society, acting, for instance, as the treasurer of the board (1955–1977). she was a long-time member of the committee of botany, polish academy of sciences, and became the polish correspondent of the committee for mapping of macromycetes in europe in 1960. in 1966, she organised the fourth congress of european mycologists in warsaw, encouraging the development of professional contacts between polish researchers of fungi, still very few at the time, and those from abroad. skirgiełło established the mycological section of the polish botanical society in 1956 and presided over it almost until her death. research meetings, seminars and trips organized by her encouraged the exchange of information and brought the academic community closer together. the section met regularly in warsaw and, as the circle of mycologists in poland grew, in wrocław, łódź, kraków, poznań and lublin, as well as during successive conventions of the polish botanical society. she also founded and was a long-standing editor of two publications, important for the polish academic world: the series mycota within the flora of spore plants in poland and the journal acta mycologica. she advocated the idea of a multi-volume study devoted to the entire polish mycoflora with contributions by polish mycologists as early as after the war. 114 t. majewski wishing to investigate poorly examined parts of poland, she organized field trips for the few polish mycologists, collected, together with her students, herbarium material for monograph studies and made every effort to attract young people interested in collaborative work. she assembled a beautiful and impressive library of mycological publications, one of the most important of this type in poland, in the department of systematics and plant geography, warsaw university. the 27 volumes of grzyby (mycota), edited by her and published within flora roślin zarodnikowych polski (the flora of spore plants in poland) in co-operation with the institute of botany, polish academy of sciences, kraków, between 1960 and 1999, was one of the fruits of her work. they provide determination keys as well as descriptions and figures of all species in a given group of fungi that had been reported from and could be found in poland. skirgiełło did her best to encourage potential contributors, was the research editor of all volumes and the author or co-author of six. three volumes were published in english. acta mycologica, founded by skirgiełło in 1965, meant much for the development of mycological studies in poland. it was initially published annually and later on biannually while skirgiełło remained editor-in-chief until 2001. the journal has played a key role as an international print forum and a place of research exchange for polish mycologists of all specializations as well as for lichenologists. skirgiełło published as many as 150 studies, including 80 research works dealing mostly with mycology as well as with paleobotany and the history of botany. macromycetes were her particularly favourite research interest. monograph studies on the polish species of the boletales (1960), cap aphyllophorales (1967), russula (1991), lactarius (1998) and pluteaceae (1999), are her most influential publications on these fungi. a talented painter, skirgiełło illustrated the books with her own colour figures of fruitbodies. skirgiełło was the author of several descriptions of rare macromycete species and their lists from underexplored areas where, as has been mentioned, she organized special research trips. she published a series of works on the distribution of higher fungi in europe between 1965 and 1986. she also edited and published (1961-1980) a series of studies on the brown-coal flora in an opencast mine in turów, including fungi, and, in conjunction with hanna czeczott, flowering plants. a dedicated and conscientious teacher, skirgiełło for many years lectured in botany, especially mycology, and the history of botany, at warsaw university. some of her publications were addressed specifically to students and lovers of fungi. she was the author of an invaluable textbook for the determination of lower fungi (grzyby niższe, 1954), which was of great help to young mycologists interested in these fascinating organisms that were still poorly examined in poland at the time. she prepared an extensive chapter on fungi in the academic textbook rośliny zarodnikowe (spore plants, four editions 1960-1986) and compiled a bibliography of polish mycological publications (1988) as well as many biographies of polish and foreign botanists or mycologists. numerous essays, brochures and boards that she designed or made for schools hoped to advance the knowledge of fungi. she also worked with the editorial boards of general and specialist encyclopaedias, translated botanical works into polish, including mykologie, grundriss für naturwissenschftler und mediziner by e. müller and w. loeffler (zarys mikologii dla przyrodników i lekarzy), two editions 1972, 1987, organized numerous exhibitions of edible and poisonous fungi, offered professor dr alina skirgiełło (1911-2007) 115 advice and assistance to many institutions and commissions as an expert mycologist. an active participant of international symposia and conferences, skirgiełło was the only person to have attended all congresses of european mycologists, from the first in brussels in 1956 to the fourteenth in yalta in 2003. although absent at the 15th congress in st. petersburg in september 2007, only one month before her death, skirgiełło submitted a joint abstract with her students. her presence at the congresses was greatly appreciated: she was vice-president in copenhagen (1970), avignon (1974) and budapest (1978), and president in bologna (1981) and oslo (1985). she also took part in the first four international mycological congresses, participated in many conventions of mycologists from germany, czechoslovakia, hungary and the baltic states, and attended the x-ème congrès international sur la science et la culture des champignons comestables (france, 1978). her notes and impressions were always made available to polish botanists at research symposia and in published reports in wiadomości botaniczne (botanical news). she was a member of the international mycological association and of the european mycological association. in 2004 she became a honorary member of the european mycological association. she received several medals, orders and rewards, including a honorary membership of the polish botanical society and the committee of botany, polish academy of sciences. her surname is remembered in the name of fungal genera and species: skirgiellia batko (1978), skirgiellopsis batko (1978), laboulbenia skirgielloae balazuc (1975) and urocystis skirgielloae piątek (2006). professor alina skirgiełło has left behind many disciples in all research and academic centres across poland. polish mycologists many of whom studied under her supervision owe her a great deal. her impressive organizational skills, catching professional enthusiasm and remarkable ability to inspire interest in mycology stimulated her colleagues and students. she founded and was editor of publications that have become important places of scholarly discussion. her presence in the academic life will be long missed. bibliography of publications by alina skirgiełło 1939. polskie naziemne grzyby rurkowe [boletaceae et polyporaceae terrestres de pologne]. planta po-planta polonica 8 (3): 1-2 nlb., 1-124, 8 pls. (in polish with french summary). 1946. nowe gatunki grzybów wyższych dla flory polskiej [new species of ascoand basidiomycetes for the polish flora]. acta societatis botanicorum poloniae 17 (1): 53-60 (in polish with english summary). – przyczynek do znajomości flory mikologicznej okolic kuźnicy grodzieńskiej [a contribution to the knowledge of ascoand basidiomycetes from the environs of kuźnica grodzieńska]. acta societatis botanicorum poloniae 17 (2): 239-251 (in polish with english summary). 1950. rodzaj russula w polsce [le genre russula en pologne]. towarzystwo naukowe warszawskie, sprawozdania z posiedzeń wydziału iv nauk biologicznych 41 (1947-1948): 7-8 (in polish with french summary). 1951. flora mikologiczna doliny łomny (czechosłowacja) [flore mycologique de la vallée łomna (tchécoslovaquie)]. acta societatis botanicorum poloniae 20 (2) (1950): 689-708 (in polish with french summary). 1952. rodzaj russula w polsce i w krajach przyległych [le genre russula en pologne et dans les pays limitrophes]. planta polonica 9 (1) (1951): 1-130, 4 pls., 1 map (in polish with french summary). 116 t. majewski nowe stanowiska pisolithus arenarius alb. et schw. w okolicach warszawy [a new stations of pisolithus arenarius alb. et schw. in the neighbourhood of warsaw]. acta societatis botanicorum poloniae 21 (3): 443-446 (in polish with english summary). 1954. grzyby niższe. pragrzyby i glonowce. przewodnik morfologiczno-systematyczny z kluczami do oznaczania [lower fungi, archimycetes and phycomycetes. characteristics and keys for determi-characteristics and keys for determination]. p.w.n., warszawa, 247 pp. 1956. helvellella sphaerospora (peck) imai en pologne. (in:) première session européenne de mycologie. belgique, 15-22 septembre 1956. programme et resumes des communications, p. 18. 1957. (skirgiełło a., nespiak a.) erfahrungen mit dermocybe orellana (fr.) in polen. a. cortinarius (dermocybe) orellanus fr. non quél. – cause d’intoxications fongiques en pologne en 1952-1955. zeitschrift für pilzkunde 23 (3-4): 138-139. – helvellella sphaerospora (peck) imai en pologne. acta societatis botanicorum poloniae 26 (2): 309-317. 1958. (skirgiełło a., nespiak a.) cortinarius (dermocybe) orellanus fr. non quél. – przyczyną licznych zatruć grzybowych w polsce w latach 1952-55 [cortinarius (dermocybe) orellanus fr. non quél. – cause d’intoxications fongiques en pologne en 1952-55]. acta societatis botanicorum poloniae 27 (2): 215-220, 1 pl. (in polish with french summary) 1959. notatki mikologiczne z okolic krościenka nad dunajcem [mycological notes from the neighbourhood of krościenko on dunajec]. monographiae botanicae 8: 229-235 (in polish with english summary). (skirgiełło a., czeczott h.) flora kopalna turowa koło bogatyni ii (1) [the fossil flora of turów near bogatynia ii (1)]. hamamelidaceae, nymphaeaceae, sabiaceae, viciaceae, nyssacae. prace muzeum ziemi 3: 93-112 (polish text), 121-128 (english text), pl. xv-xx. 1960. de la nécessité de la protection des champignons et des derrains respectifs. (in:) druhý sjezd evropských mykologů, československo 1960, p. 25. – borowikowe (boletales). (in:) flora polska. grzyby [t . 1] [flora of poland, fungi, vol. i]. inst. bot. pan, pwn, warszawa, pp. 131, 30 pls. (in polish). – wiosenne miseczniaki białowieży [discomycètes de printemps de białowieża]. monographiae botanicae 10 (2): 3-19 [in polish with french summary). – (domański s., gumińska b., lisiewska m., nespiak a., skirgiełło a., truszkowska w.) mikoflora bieszczadów zachodnich (wetlina 1958) [mycoflore des bieszczady occidentales (wetlina 1958)]. monographiae botanicae 10 (2): 159-237 (in polish with french summary). 1961. flora kopalna turowa koło bogatyni ii (2) [the fossil flora of turów near bogatynia ii (2)]. ascomycetes, basidiomycetes. prace muzeum ziemi 4: 5-12 (polish text), 85-88 (english text), pl. i-iv. – (czeczott h., skirgiełło a.) flora kopalna turowa koło bogatyni ii (2) [the fossil flora of turów near bogatynia ii (2)]. juglandaceae. prace muzeum ziemi 4: 51-73 (polish text), 103-113 (english text), pl. xvi-xxi. – (czeczott h., skirgiełło a.) flora kopalna turowa koło bogatyni ii (2) [the fossil flora of turów near bogatynia ii (2)]. aceraceae. prace muzeum ziemi 4: 78-81 (polish text), 116-117 (english text), pl. xxi. – de quelques champignons supérieurs récoltés par m. kuc au spitsbergen en 1958. bulletin of the research council of israel, sect. d, 10: 287-293. – de la nécessité de la protection des champignons et des terrains respectifs. česka mykologie 15 (3): 153-158. – (podbielkowski z., rejment-grochowska i., skirgiełło a.) rośliny zarodnikowe [cryptogamous plants]. p.w.n., warszawa, 970 pp. (ii ed. 1979 and 1980; iii ed. 1982; iv ed. 1986) (in polish). – tertiärpilze aus der grube turow. zeitschrift für pilzkunde 27 (2-4): 90-93. 1963. (domański s., gumińska b., lisiewska m., nespiak a., skirgiełło a., truszkowska w.) miko-mikoflora bieszczadów zachodnich. ii (ustrzyki górne, 1960) [mycoflora of west bieszczady. ii]. monographiae botanicae 15: 3-75 [in polish with english summary). – (skirgiełło a., rudnicka-jezierska w.) nowe stanowiska dwóch interesujących wn�trzniaków (gas-nowe stanowiska dwóch interesujących wn�trzniaków (gasteromycetes) w polsce [new stations of two interesting gasteromycetes in poland]. monographiae botanicae 15: 355-360 [in polish with english summary). – (skirgiełło a., wosińska a.) o rozmieszczeniu jeleniaków (elaphomyces) w polsce [elaphomyces distribution in poland]. monographiae botanicae 15: 361-371 [in polish with english summary). professor dr alina skirgiełło (1911-2007) 117 1965. materiały do poznania rozmieszczenia geograficznego grzybów wyższych w europie. i. xerocomus parasiticus (bull. ex fr.) quél. i pycnoporus cinnabarinus (jacq. ex fr.) karst. w polsce [matériaux à la connaissance de la distribution géographique des champignons supérieurs en europe. i] . acta mycologica 1: 23-26 [in polish with french summary). 1966. czwarty kongres europejskich mikologów. quatrième congrès des mycologues européens. po-quatrième congrès des mycologues européens. pologne (...). guide. warszawa, pp. 1-98, i-v, 11 figs. [four language issues; a. skirgiełło was the co-author and editor]. 1967. (domański s., orłoś h., skirgiełło a.) bezblaszkowe (aphyllophorales), żagwiowate ii (polyporaceae pileatae), szczeciniakowate ii (mucronoporaceae pileatae), lakownicowate (ganodermataceae), bodarcewowate (bondarzewiaceae), boletkowate (boletopsidaceae), ozorkowate (fistulinaceae). (in:) flora polska. grzyby, t. 3 [flora of poland, fungi, vol. 2]. inst. bot. pan, pwn, warszawa, pp. 398, 29 pls. (in polish). – materiały do poznania rozmieszczenia geograficznego grzybów wyższych w europie. ii [matériaux à la connaissance de la distribution géographique des champignons supérieurs en europe. ii]. acta mycologica 3: 243-249 [in polish with french summary). – (domański s., gumińska b., lisiewska m., nespiak a., skirgiełło a., truszkowska w.) mikoflora bieszczadów zachodnich. iii (baligród, 1962) [mycoflora of west bieszczady. iii]. acta myco-acta mycologica 3: 63-114 [in polish with english summary). – (czeczott h., skirgiełło a.) flora kopalna turowa koło bogatyni ii(3) [the fossil flora of turów near bogatynia ii (3)]. monocotyledones, araceae; dicotyledones: betulaceae, menispermaceae, meliaceae, passifloraceae, combretaceae, trapaceae, symplocaceae, styraceae. prace muzeum zi-prace muzeum ziemi 10: 98-115, 122-141 (polish text); 143-152, 156-166 (english text), pl. i-v, vii-x. 1968. higher fungi collected in 1958 at hornsund, westspitsbergen. (in:) k. birkenmajer (ed.), polish spitsbergen expeditions 1957-1960. summary (...). wydawnictwa geologiczne, warszawa, p. 113116, pl. i. – [skirgiełło a.] compte-rendu du iv-ème congrès des mycologues européens warszawa 1968 [recte: 1966]. acta mycologica 4 (2): 181-198, 2 pls. 1970. materiały do poznania rozmieszczenia geograficznego grzybów wyższych w europie. iii [matéri-iii [matériaux à la connaissance de la distribution géographique des champignons supérieurs en europe. iii]. acta mycologica 6 (1): 101-123 (in polish with french summary). – (skirgiełło a., g�sicka i.) riessia semiophora w polsce [riessia semiophora en pologne]. acta mycologica 6 (1): 125-127 [in polish with french summary). – (domański s., lisiewska m., majewski t., skirgiełło a., truszkowska w., wojewoda w.) mikoflora bieszczadów zachodnich. iv (zatwarnica, 1965) [mycoflora of west bieszczady. iv]. acta myco-acta mycologica 6 (1): 129-179 [in polish with english summary). 1972. certains problèmes du domaine de la géographie des champignons. mycopathologia et mycologia applicata 48 (1): 87-91. – materiały do poznania rozmieszczenia geograficznego grzybów wyższych w europie. iv [matériaux à la connaissance de la distribution géographique des champignons supérieurs en europe. iv]. acta mycologica 8 (2): 191-218 (in polish with french summary). 1973. (domański s., orłoś h., skirgiełło a.) fungi. polyporaceae ii (pileatae), mucronoporaceae ii (pileatae), ganodermataceae, bondarzewiaceae, boletopsidaceae, fistulinaceae. translated from polish. u.s. department of agriculture (...). washington, warsaw, pp. 332, 27 pls. 1974. (ciborska e., sadowska b., skirgiełło a., zadara m.) wst�pne badania nad mikoflorą łąk nawożonych [contribution to the knowledge of mikoflora (!) presence on fertilized pastures]. roczniki nauk rolniczych, ser. e, 4 (2): 173-176 (in polish with english summary). 1975. 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(in:) xv congress of european mycologists, saint petersburg, abstracts, p. 84-85. tomasz majewski 2014-01-01t11:47:38+0100 polish botanical society wood-inhabiting fungi on pedunculate oak coarse woody debris in relation to substratum quantity and forest age reda iršėnaitė1 and ernestas kutorga1, 2 1laboratory of mycology, institute of botany žaliųjų ežerų 49, lt-08406 vilnius, reda.irsenaite@botanika.lt 2department of botany and genetics, faculty of natural sciences, vilnius university m.k. čiurlionio 21/27, lt-03101 vilnius, ernestas.kutorga@gf.vu.lt i r š ė n a i t ė r., k u t o r g a e.: wood-inhabiting fungi on pedunculate oak coarse woody debris in relation to substratum quantity and forest age. acta mycol. 42 (2):169-178, 2007. wood-inhabiting fungi on pedunculate oak (quercus robur) coarse woody debris (cwd) was investigated in 50 plots of 0.1 ha in oak stands of different ages in lithuania. in maturing stands (50-120 years) the average volume of oak cwd was 4.7 m3/ha, and in mature stands (over 120 years) – 13.9 m3/ha. both in maturing and mature stands, the greatest fraction of cwd consisted of fallen oak branches (81 % and 84 % respectively), whereas fallen trunks comprised about 10 % of the total units of cwd. in total 1350 records of 203 species (49 ascomycetes and 154 basidiomycetes) were collected during 2 years of investigation. species richness and abundance increased significantly with the increase of volume and abundance of cwd. higher species richness was detected in mature stands than in maturing ones. woodinhabiting species composition varied greatly at stand scale, and one third of all detected species occurred only in one plot. red-listed fungi were found only in mature stands. we conclude that, even in managed oak stands, oak cwd maintains a rather diverse species composition of ascomycetes and basidiomycetes. however, the current practice of forestry in lithuania of removing dying or dead wood of large volume, e.g. standing and fallen trunks, reduce the distribution of highly specialized, usually rare, and endangered fungi. key words: xylotrophic fungi, coarse woody debris, quercus robur, fungal conservation introduction large pieces of dead rotting fallen trunks, branches and stumps, which characterize coarse woody debris (cwd), are substantial components in forest ecosystems (h a r m o n et al. 1986). there is no doubt that woody debris plays an essential role in creating habitats for many saproxylic organisms, including wood-inhabiting fungi (b o d d y 2001; s i i t o n e n 2001). the diversity and frequency of wood-inhabiting fungi depend on the stage of wood decay (n i e m e l ä , r e n v a l l , p e n t i l l ä 1995; acta mycologica vol. 42 (2): 169-178 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 170 r. iršėnaitė and e. kutorga r e n v a l l 1995; l i n d b l a d 1998) and in general considered to be greater on large diameter woody debris (b a d e r , j a n s s o n , j o n s s o n 1995; h ø i l a n d , b e n d i k s e n 1997; k r u y s et al. 1999). recent studies indicate that forest management and fragmentation in boreal coniferous forests have greatly affected the diversity of wood-inhabiting fungi, especially the occurrence of rare species (s i p p o l a , r e n v a l l 1999; l i n d g r e n 2001; p e n t t i l ä , s i i t o n e n , k u u s i n e n 2004). studies dealing with temperate broadleaved forests have also analyzed the relationship between fungal species diversity (both basidiomycetes and ascomycetes) and woody debris quantity/ quality (r u n g e 1975; l a n g e 1992; h e i l m a n n c l a u s e n , c h r i s t e n s e n 2004; n o r d é n et al. 2004), some environmental factors (h e i l m a n n c l a u s e n 2001), and stand structure/age (n o r d é n , p a l t t o 2001). as in most parts of europe, the old-growth temperate broadleaved forests in lithuania have been fragmented to rather small fractions (p e t e r k e n 1996; k e n s t a v i č i u s 1997), and intensive forest management has endangered many organisms, including fungi, associated with oak (s u n h e d e , va s i l i a u s k a s 1996, 2003). out of total 40 red-listed wood-inhabiting fungi in lithuania (a n o n y m o u s 2005), more than 10 species are confined to the pedunculate oak (quercus robur), both ecologically and economically important tree species in lithuania. quercus robur stands occupy about 33600 ha, i.e. 1.8 % of the total forest area in lithuania (n a v a s a i t i s et al. 2003), and they usually grow intermixed with other deciduous or coniferous trees. the main focus of most mycological studies on oak wood in lithuania has been species composition and distribution, and the quantity and quality of cwd has not usually been considered. moreover, there is no up to date and generally available information on the amount of woody debris in lithuanian forests. the first attempt to examine fungi and cwd in various coniferous and deciduous unmanaged stands of 40-130 years old is that of va s i l i a u s k a s et al. (2004). they found the volume of cwd of different tree species to be larger in older stands than in younger ones, demonstrated the variation of structural characteristics of cwd, and recorded 41 species of wood-decomposing polypores. nevertheless, their sampling of oak cwd was too limited to draw conclusions about distribution patterns, and the oak wooddecomposing fungi were not specified. therefore, in this study we examined the present status of cwd of oak and the diversity of fungi associated with it. the study focuses on wood-inhabiting ascomycetes and basidiomycetes on coarse woody debris in managed oak dominated stands of different age. the list of recorded species and the analysis of fungal composition on wood of different decay stage are presented in separate paper (i r š ė n a i t ė , k u t o r g a 2006). the aim of this paper was to analyse the species richness of oak wood-inhabiting fungi with respect to quantitative and structural features of cwd in maturing and mature oak stands. material and methods study sites and plots. ten study sites were selected in different parts of lithuania (fig. 1) according to the forest management documents and maps. study sites 1-2, 5-8 and 10 comprised of forests growing on mezotrophic humid soil and dominated by pedunculate oak (quercus robur). the co-existing tree species are picea abies, wood-inhabiting fungi 171 populus tremula, tilia cordata and fraxinus excelsior. the bush layer is moderately thick and consists mainly of corylus avellana, lonicera xylosteum, frangula alnus and sorbus aucuparia. sites 3 and 4 comprised forests on mezotrophic dry soil, dominated by both quercus robur and pinus sylvestris, and with a sparse shrub layer of corylus avellana and sorbus aucuparia. site 9 is an abandoned oak-wood pasture with solitary corylus avellana. local foresters´ information and cut stumps we observed indicate that all sites were more or less managed during the last 50 years, mostly by thinning or removing dead trees. the study comprised two oak stand age groups: i, – maturing (50–120 years); and ii, – mature (over 120 years). according to the study site size, from one to nine circular study plots of 0.1 ha were randomly selected. in total, we surveyed 50 plots (25 in each age group). examination of cwd. the method for the examination of cwd was prepared according to k r u y s et al. (1999). within each plot, all fallen and standing dead oak wood with the base 5 cm in diameter and more and over 30 cm in length was investigated. the following characteristics were recorded for each unit of dead wood that harboured a fungus: category of cwd (lying trunk, snag, stump, branch), maximum (base) and minimum (top) diameter (m), length (m), and decay stage (ds). five cwd decay stages were defined following a modified classification of r e n v a l l (1995). if the stage of decomposition varied in different parts of a trunk, an average decay stage was considered. cwd volume was calculated by the smalian’s formula (p e t t e r s o n , w i a n t , wo o d 1993). sampling and identification of fungi. the fieldwork was undertaken in 2001– 2002 during the main period of fruit body production (august–october) in both those years, with a single visit at each plot (up to 3 hours). both basidiomycetes and ascomycetes were inventoried. recorded species were arranged into these fungal groups: discomycetes, pyrenomycetes and loculoascomycetes, agarics (including cyphelloid fungi), corticioid fungi (including thelephoroid fungi), gasteromycetes, heterobasidiomycetes, polypores (including ramaria). these commonly used fungal groups are based on fruit body types, and are not taxonomical groups reflecting phylogenetic relationships. a species found on a single unit of wood was considered as fig. 1. the location of the study sites in lithuania: 1, kreiviškės; 2, plikoji sala; 3, minijos kilpa; 4, šilas; 5, gojus; 6, drausgiris; 7, subartonys; 8, ąžuolija; 9, ginučiai; and 10, dūkštos. 172 r. iršėnaitė and e. kutorga one record, regardless of the number of observed fruit bodies. easily recognizable species were identified in the field, and about 1000 specimens were collected for microscopic examination. voucher specimens are preserved in the herbarium of the institute of botany, vilnius (bilas). data analysis. differences in number of species and abundance and the amount and volume of cwd in different age stands were tested for significance using a non-parametric mann-whitney u-test. cwd profile was quantified by summarizing the volume of cwd in a 2-way cross-table of maximum diameter and decay stages (s t o k l a n d 2001). the chi-squared test was used to examine differences in the distribution of fungal groups and cwd decay stages. the shannon diversity index (h’) was used to evaluate the diversity of decay stages of cwd in the study plots. the first-order jackknife estimator was used to calculate true species richness (z a r 1999). dissimilarity in species composition between study plots in one study site was calculated with the sørensen distance measure, and spearman’s rank correlation coefficient (rs) was used to express the correlation between different variables. all statistical analyses were run using statistica 6.0 (s t a t s o f t i n c 2001) and pcord 4.0 software (m c c u n e , m e f f o r d 1999). results characteristics of cwd. in total 321 units (46.5 m3) of oak coarse woody debris were recorded across all study plots, taking both years together. the mature stands had significantly more cwd (203 units; 34.8 m3) than the maturing ones (118 units; 11.7 m3) (mann-whitney u-test; p<0.001). there was large variation in cwd volume within plots, from 0.01 m3 to 5.61 m3 in maturing stands, and from 0.25 m3 to 7.12 m3 in mature stands. data on average characteristics of cwd are given in table 1. the chi-squared test indicates that volume of cwd of different decay stage is distributed in the same proportions in maturing and mature stands (χ2 = 4.62; df = 4; α = 0.05). the cwd patterns with respect to dimensions and decay stages shown in table 2, 3. fallen branches comprised the largest proportion of the total units of cwd, both in maturing (81 %) and in mature stands (84 %). the proportion of lying oak trunks was respectively only 9 % and 10 %. stumps and snags formed the minority of all recorded cwd. species richness. in total 1350 records of 203 species (49 ascomycetes and 154 basidiomycetes) were collected over the period of investigation. four species (2 %) ta b l e 1 characteristics of the cwd, the number of records and fungal species (mean ± standard deviation) in study plots stand age group n no. of cwd units per ha diamc of cwd (m) volume of cwd (m3/ha) no. of records per ha no. of species per ha i a 25 47.2 (±17.7) 0.17 (±0.12) 4.7 (±11.4) 199.6 (±98.1) 157.2 (±73.7) iib 25 81.2 (±27.1) 0.22 (±0.07) 13.9 (±15.8) 338.0 (±107.2) 262.8 (±81.8) explanations: a maturing stands; b mature stands; c base diameter of the cwd unit. wood-inhabiting fungi 173 were found in more than half of the study plots, while 67 species (33 %) were found only in one. the most frequently encountered species (more than 30 records) were eriopezia caesia, humaria hemisphaerica, hyphodontia quercina, hymenochaete rubiginosa, mollisia cinerea, nemania serpens, phanerochaete velutina and schizopora paradoxa. two species, fistulina hepatica and xylobolus frustulatus, listed in the lithuanian red data book (a n o n y m o u s 2005) were also found. there was large variation in species numbers within plots in maturing stands (range 7 to 37 species) and in mature stands (range 11 to 43), but the number of species in mature stands (167) was significantly higher than in the maturing ones (134) (mann-whitney u-test; p = 0.001). the mean number of species and records per ha in study plots are shown in table 1. thirty-six species were exclusive to maturing stands, and 69 to mature stands. nearly half (48 %) of all species were common to stands of both age groups. the total number of records made was significantly greater in mature stands than in maturing ones (mann-whitney u-test; p<0.001), but a comparison of distribution proportions of the species in the different fungal groups in maturing and mature stands revealed similar patterns of occurrence (χ2 = 11.5; df = 6; α = 0.05). the number of records of corticioid fungi and discomycetes in mature stands were almost twice as high as that in maturing ones, although the number of species in both stands were nearly the same (tab. 4). polypores, gasteromycetes and heterobasidiomycetes were much less abundant in comparison with other fungal groups. as shown in table 5, species composition varied greatly between plots even within a single study site (dūkštos forest), but it was more similar in plots of the same age group. ta b l e 2 the volume of cwd (m3 per ha) in maturing stands in a cwd profile based on their maximum (base) diameter and decay stage d ec ay s ta ge i 0.05 ii 0.02 0.14 0.03 0.02 0.09 0.02 iii 0.04 0.16 0.10 0.15 0.15 iv 0.06 0.35 0.38 0.19 0.58 0.05 1.95 v 0.01 0.03 0.02 5 10 20 30 40 50 60 70 80 90 100 110 120 base diameter (cm) ta b l e 3 the volume of cwd (m3 per ha) in mature stands in a cwd profile based on their maximum (base) diameter and decay stage d ec ay s ta ge i 0.02 0.03 ii 0.01 0.07 0.16 0.07 0.02 iii 0.08 0.88 0.64 0.31 1.66 0.21 1.61 0.09 0.09 iv 0.09 0.74 0.62 1.33 2.01 0.99 0.92 0.05 0.39 v 0.01 0.50 0.09 0.03 0.11 0.12 5 10 20 30 40 50 60 70 80 90 100 110 120 base diameter (cm) wood-inhabiting fungi 175 the estimation of species richness by the jackknife estimator indicated that the actual number of species present should be higher than those recorded. in plots of age group i and ii, 70 % and 73 % respectively of the estimated number of species were recorded. patterns of relationships. significant positive correlations were found between the abundance and richness of fungal species and cwd variables (tab. 6). the number of species and records showed a higher correlation with the volume of cwd than with the number and average diameter of cwd units. no significant correlation between diversity of decay stages in the study plots and species abundance was revealed. however, there was a significant correlation between the mean dbh of living oaks, an expression of the oak’s age, and the variables measured. discussion coarse woody debris of pedunculate oak. in lithuania the average volume of quercus robur cwd in mature oak stands (13.9 m3/ha) is three times higher than in maturing ones (4.7 m3/ha). this corresponds closely with the volume of cwd in switzerland: 13.8 m3/ha in natural oak forests, and 5.3 m3/ha in managed ones (b r e t z , g u b y and d o b b e r t i n 1996). abandoned oak wood-pastures with high biodiversity conservation value in sweden are on average 100-160 years old and contain only 5 m3/ha of coarse and 1.9 m3/ha of fine oak woody debris (n o r d é n et al. 2004). the average volume of cwd of managed mature deciduous forests in england (12–23 m3/ha) and poland (10 m3/ha) is comparable to that of mature forests in lithuania, but the calculated average volume of wood debris of natural deciduous forests in poland is several times higher (94 m3/ha) (k i r b y , we b s t e r , a n t c k z a k 1991). comparison of these data shows that different amounts of cwd are affected by forest type, age, and management intensity. the important feature of our study plots was the abundance of fallen oak branches and the lack of large dead trunks. the volume of cwd of various decay stages was distributed in the same proportions in mature and maturing stands, and in both stands low cwd abundance and multiply gaps were observed from cwd profile. the extremely small amount of recently fallen woody debris (< 1 m3/ha) and very small amount of large diameter and strongly decayed wood indicates constant management in oak stands. fungal species richness and composition. the diversity of wood-inhabiting species on oak cwd in our study plots was high, and according to jackknife estimator it could be one third more in stands of both age group. to f t s and o r t o n (1998) stated that investigation of all species is hardly achievable in mycological inventories, unless the investigated community is very homogenous, as in the corticioid fungal community on alnus glutinosa (k ü f f e r , s e n n i r l e t 2000). most recorded species are not confined to oak wood, but can also fructify on wood of other deciduous trees, and to a lesser extent also on wood of coniferous trees. some of them, like tomentelloid fungi, are not saprotrophs but ectomycorrhizal species, using oaks wood only as living habitat. comparison of taxonomical structure of fungi on oak debris detected in this study with other related data is difficult because different sampling methods. l i n d h e , å s e n b l a d and to r e s s o n (2004) investigated only cut quercus robur logs and reported that the logs harboured 53 fungal species, where 176 r. iršėnaitė and e. kutorga the agarics comprised 45 %, corticioid fungi 13 %, and polypores 11 %. our results (47 % of corticioid) are consistent with the data on fungi on coarse and fine woody debris in sweden (n o r d é n et al. 2004), where the corticoid fungi also predominated (50 %). comparison of maturing and mature stands. the number of species and occurrence rate of wood-inhabiting fungi correlated positively not only with the cwd amount and volume, but also with dbh of living oak, as expression of stand age. therefore, significantly more abundance of species was recorded in mature stands than in maturing ones. with a larger volume of cwd, higher numbers of woodinhabiting fungal species could be expected. thus, if in maturing stands the volume of cwd was larger the greater species richness of wood-inhabiting fungi would be expected. despite the proportions of fungal groups being similar in stands of different age groups, the differences in species composition between mature and maturing plots as well as between plots of same age group were obvious. the number of unique species in mature stands was twice as high as in maturing ones, and redlisted species were found only in mature stands, an observation consistent with other studies showing that the red-listed wood-inhabiting fungi prefer old-growth forests (s i p p o l a , r e n v a l l 1999; p e n t t i l ä et al. 2004; va s i l i a u s k a s et al. 2004) and woody debris of large diameter (r e n v a l l 1995; k r u y s et al. 1999; l i n d h e et al. 2004). in our study the occurrence of two red-listed species only in mature stands can be explained by the preference of fistulina hepatica to large trunks or stumps, and of xylobolus frustulatus to coarse branches or fallen trunks of old oaks. other red-listed species, such as inonotus dryadeus, i. dryophilus, piptoporus quercinus, grifola frondosa and hapalopilus croceus, not found during our research are mostly confined to large old living oak trunks or recently fallen ones. perenniporia medullapanis, specific to old oak stumps or snags, was also not recorded in study plots. this large diameter woody debris was remarkably scarce in both maturing and mature stands, which may have determined the low proportion of not only red-listed species, but of common polypores detected. conclusions our results confirm that wood-inhabiting fungal species richness depends on the amount of cwd. the average amount of cwd in managed mature oak stands in lithuania is 13.9 m3/ha. in accordance with similar studies (k i r b y et al. 1991) it is evidently advisable to maintain not less than 20 m3/ha of woody debris in temperate broadleaved stands. fallen branches comprise the largest fraction of oak cwd and greatest diversity of fungi species. however, large fallen trunks should be maintained in stands as well, because this valuable substrate is usually heterogeneous and comprises less or more decayed parts, and remnants of bark and branches; such substrate is especially important for rare polypores. we conclude that, even in managed oak stands, oak cwd maintains a rather diverse species composition of ascomycetes and basidiomycetes. although in general the highest species richness was detected in mature stands in our study, the species richness more correlated with the amount of cwd then with stand age itself. wood-inhabiting fungi 177 some maturing stands with a high amount of cwd, there were nearly as many or even more fungi than in particular mature stands. however, a very small amount of freshly fallen wood and strongly decayed woody debris, especially of large diameter trunks, and gaps in cwd continuity both in mature and maturing stands indicates long lasting management practices in lithuanian oak stands, which reduce the distribution of highly specialized, usually rare, and endangered fungi. acknowledgements. we thank nils hallenberg (gothenburg) and urmas kõljalg (tartu) for help in determining various critical specimens. we are also grateful to david l. hawksworth (madrid) for the suggestions to improve the manuscript and linguistic revision of the manuscript. the study was financially supported partly by the lithuanian state science and studies foundation and the swedish institute. references a n o n y m o u s 2005. dėl aplinkos ministro 2003 m. 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(in:) b.g. j o n s s o n , n. k r u y s (eds). ecology of wood debris in boreal forests. ecol. bull. 49: 71−83. blackwell science, oxford. s u n h e d e s., va s i l i a u s k a s r. 1996. wood and bark inhabiting fungi on oak in lithuania. baltic forestry 2: 23−27. s u n h e d e s., va s i l i a u s k a s r 2003. hotade tickor på ek i litauen. svensk bot. tidskr. 97: 252−265. to f t s r.j., o r t o n p.d. 1998. the species accumulation curve for agarics and boleti from caledonian pinewood. mycologists 12: 98−102. va s i l i a u s k a s r., va s i l i a u s k a s a. s t e n l i d j., m a t e l i s a. 2004. dead trees and protected polypores in unmanaged north-temperate forest stands of lithuania. forest ecol. manag. 193: 355−370. z a r j.h. 1999. biostatistical analysis. upper saddle river, new jersey. 2014-01-01t11:46:00+0100 polish botanical society ophiostomatoid fungi isolated from fallen shoots of scots pine pruned by tomicus species in poland robert jankowiak1 and miroslav kolařík2 1agricultural university of cracow, department of forest pathology al. 29 listopada 46, 31-425 kraków, rljankow@cyf-kr.edu.pl 2institute of microbiology v.v.i, academy of sciences of the czech republic vídeňská 1083, 142 20 prague 4, czech republic jankowiak r., kolařík m.: ophiostomatoid fungi isolated from fallen shoots of scots pine pruned by tomicus species in poland. acta mycol. 46 (2): 201–210, 2011. ophiostomatoid fungi are known to be associated with tomicus spp. on pinus sylvestris. however, very little is known about the fungi present in the pine shoots damaged by these insects. the aim of this study was to survey species of ophiostoma s.l. associated with fallen shoots of scots pine pruned by tomicus spp. in poland. the study was conducted in four pure scots pine stands in southern and south-western part of the country. fungi were identified based on morphology and dna sequence comparison for two gene (the its rdna region and ß–tubulin). in total, 64 isolates obtained, represent seven species of ophiostomatoid fungi, including their asexual states of the genera. two of them, ophiostoma sp. 1 and sporothrix sp. 1 probably represent new taxa. all species were found at very low frequencies. among them, ophiostoma minus, ophiostoma sp. 1 and sporothrix sp. 1 were the most frequently isolated, with a frequency of 2.0%. occasionally, isolated species were: leptographium piriforme, ophiostoma canum, o. floccosum and grosmannia cucullata-like. association of species of ophiostoma s.l. with tomicus spp. and the taxonomic status of two new species are discussed. key words: ophiostoma, sporotrix, pinus sylvestris, new taxa, bark beetles introduction many species of bark beetles (coleoptera: scolytinae) have been reported from pine stands of poland. these insects are generally considered secondary colonizers, which attack stressed, dying or dead trees. among them, tomicus pini perda (l.) and t. minor (hart.) are able to attack healthy standing scots pine trees under favourable conditions. the damage by bark beetles of the genus tomicus in pine trees is caused by two feeding strategies. as in many species of bark beetles acta mycologica vol. 46 (2): 201–210 2011 202 r. jankowiak and m. kolařík one strategy is penetration of the protective bark and nutritive phloem of tree stems, and transmission of blue-stain fungi in the sapwood. another strategy occurring only in tomicus beetles is penetration and maturation feeding of adults in living shoots of healthy trees. the shoots become hollow and easily break off. during this period of maturation-feeding, each new adult feeds on the currentyear or last year’s shoots, mostly in the upper half of the crown. adults bore into the bark of shoots and hollow out the pitch to a length of 2-3 cm. at strong winter, most of the injured shoots break off and fall to the ground. when shoot feeding is severe, tree height and diameter growth are reduced (borkowski 2001). species described in the past in the genus ophiostoma s.l. represent a group of morphologically similar genera characterized by ascomata with elongated perithecial necks. in fact, according to the recent taxonomic studies most of them belong to the genus ophiostoma syd.& p. syd. s. str. with pesotum j.l. crane & schokn. and sporothrix hektoen & c.f.perkins anamorphs, grosmannia goid. with leptographium lagerb. & melin anamorphs and ceratocystiopsis h. p. upadhyay & w. b. kendr. with hyalorhinocladiella h.p. upadhyay & w.b. kendr. anamorphs (upadhyay 1981; zipfel et al. 2006). these genera and the members of the genus ceratocystis ellis & halst. s. str. have been referred to as the ophiostomatoid fungi (wingfield et al. 1993). ophiostomatoid fungi are the predominant associates of tomicus latreille in europe. specific relationships between fungi and tomicus spp. are the most clear for t. minor. the ambrosia fungus, ambrosiella tingens (lagerb. & melin) l.r. batra and ophiostoma canum are common and consistent associates of t. minor in europe (mathiesen 1950, 1951; francke-grosmann 1952; mathiesen-käärik 1953; kotýnková-sychrová 1966; kirisits et al. 2000; jankowiak 2008). in the recent studies, the association of t. minor also with ophiostoma minus, o. piceae (münch) syd. & p. syd., leptographium procerum (w.b. kendr.) m.j. wingf., l. lundbergii lagerb. & melin (jankowiak 2008) and ophiostoma canum-like species (linnakoski et al. 2010) has been reported. the association between t. piniperda and ophiostomatoid fungi is looser because this insect carries numerous fungal species, but with low and inconsistent frequency. among them, o. minus and leptographium wingfieldii m. morelet were dominant fungal species in many populations in europe (lieutier et al. 1989; gibbs, inman 1991; solheim, långström 1991; wingfield, gibbs 1991; jankowiak 2006; jankowiak, kurek 2006; jankowiak, bilański 2007). although tomicus spp. mycobiota is relatively well recognized in europe (kirisits 2004), we have no information about fungal taxa associated with pine shoots damaged by these insects. it is unknown whether shoots damaged by adult individuals feeding in scots pine crowns are colonized by fungal species carried by tomicus beetles. we often observed that numerous arthropods penetrated fallen pine shoots and therefore we presume that these organisms can act as vectors for fungal species not closely associated with tomicus spp. the studies were aimed to survey the ophiostomatoid fungi associated with pine shoots damaged by tomicus spp. collected from poland. we determined isolates to the species level using their morphological characteristics, as well as dna sequencing data. ophiostomatoid fungi 203 materials and methods isolation of fungi from pine shoots. the investigations were carried out in 20072008 in four pure scots pine stands in poland (fig. 1). in october, the fallen scots pine shoots (100 per site) damaged by beetles were gathered from the forest floor and placed in separate clean paper bags. later, shoots were stored for maximum 48 hours in a cool room at 4° c until they were used for the fungi isolation. shoot samples were cut into 5 cm long sections and surface disinfected by immersing in 95% ethyl alcohol. after drying, small pieces of shoots (about 5x5 mm) were removed from each shoot section (six pieces per one shoot) and placed in petri dishes containing a selective medium for ophiostoma spp. (20 g malt extract, 20 g agar, and 1l distilled water, amended with 0.05% cycloheximide). resulting isolates were purified by transferring mycelium from the edges of single colonies to fresh 2% mea (tab. 1). the representative strains were deposited in the culture collection of fungi, department of forest pathology, agricultural university of cracow. culture morphology and dna sequencing. isolates were initially identified and grouped based on the culture morphology. representatives of each of the groups obtained were selected for dna sequencing (tab.1). fungi were identified on the basis of morphological characteristics by comparison with published data and reference cultures deposited at department of forest pathology, agricultural university of cracow. identification based on morphology was confirmed by dna sequencing of the representative isolates (tab.1). its rdna region (its1-5.8 s-its2) were amplified using the primers its1 and nl4 as described by kolařík et al. (2006). a partial ß-tubulin sequence was determined for a subset of isolates, including an unknown ophiostoma sp. 1 and sporothrix sp. 1 isolates using the primers t1 or bt2a and bt2b (glass, donaldson 1995; o’donnell, cigelnik 1997). the amplicons were purified and both strands were sequenced using the same primers by macrogen inc. (seoul, korea). the sequences were compared with data from genbank using a blast similarity search. sequences generated in this study and sequences of other, related species from genbank were aligned using clustal w (thompson et al. 1994). phylogenetic trees were constructed with mega 5.04 using the neighbour-joining (nj) method and kimura two-parameter with a transition to transversion ratio (ti/tv=2) (tamura et al. 2011). bootstrap values were calculated for 1000 replications. fig. 1. map of sampling sites: chrośnica (15°59′36″n, 52°17′13″e); babimost (15°50′33″n, 52°09′11″e), jaroszowiec (19° 36′45″n, 50°20′18″e), mielec (21° 29′36″n, 50°18′53″e). 204 r. jankowiak and m. kolařík ta bl e 1 l is t o f s tr ai ns o f f un gi s pe ci es is ol at ed fr om p in e sh oo ts in fe st ed b y to m ic us s pp . u se d fo r d n a s eq ue nc in g an d th ei r g en b an k ac ce ss io n nu m be rs ta xo n is ol at e si te a pr im er us ed fo r am pl ifi ca ti on a cc es si on no . c lo se st m at ch in b l a st a cc es si on o f m at ch id en ti ty % g ro sm an ni a cu cu lla ta -l ik e (h . s ol he im ) z ip fe l, z .w . de b ee r & m .j w in gf . 18 3b r j c it s1 f /n l 4 jq 29 28 36 g . c uc ul la ta g u 06 77 58 .1 98 .9 l ep to gr ap hi um p ir ifo rm e g re if , g ib as & c ur ra h 21 5r j j it s1 f /n l 4 f m 99 20 31 l . p ir ifo rm e d q 88 52 43 10 0 o ph io st om a ca nu m (m ün ch ) sy d. & p . s yd . 21 9r j j it s1 f /n l 4 jq 29 28 26 o . c an um a j5 38 34 2. 1 10 0 o ph io st om a flo cc os um m at h. -k ää ri k 14 5r j 14 8r j c c it s1 f /n l 4 it s1 f /n l 4 jq 29 28 30 jq 29 28 29 o . fl oc co su m o . fl oc co su m a j5 38 34 3. 1 a j5 38 34 3. 1 10 0 10 0 o ph io st om a m in us (h ed gc .) s yd . & p . s yd . 16 8r j 19 0b r j c c it s1 f /n l 4 it s1 f /n l 4 jq 29 28 27 jq 29 28 35 o . m in us o . m in us jf 44 05 85 .1 a m 94 38 86 .1 10 0 10 0 o ph io st om a sp . 1 10 5r j 11 6r j 15 9r j 18 3a r j 18 8r j 19 8r j 23 5r j c c c c c c j it s1 f /n l 4 b t2 a/ b t2 b it s1 f /n l 4 it s1 f /n l 4 b t2 a/ b t2 b it s1 f /n l 4 b t2 a/ b t2 b it s1 f /n l 4 b t2 a/ b t2 b it s1 f /n l 4 it s1 f /n l 4 jq 29 28 33 jq 29 28 34 jq 29 28 22 jq 29 28 23 jq 29 28 24 jq 29 28 25 jq 29 28 31 o . q ue rc us ( g eo rg ev .) n an nf . o . p se do ts ug ae ( r um bo ld ) a rx o . q ue rc us o . q ue rc us o . b re vi us cu lu m w .h si n c hu ng , y am ao ka , u zn ov ic & j .j . k im o . q ue rc us o . p se do ts ug ae o . q ue rc us o . p se do ts ug ae o . q ue rc us o . q ue rc us h m 05 13 98 .1 a y 54 25 10 .1 h m 05 13 98 .1 h m 05 13 98 .1 a b 20 04 28 .1 h m 05 13 98 .1 a y 54 25 10 .1 g u 06 22 72 .1 a y 54 25 10 .1 h m 05 13 98 .1 g u 79 72 05 .1 96 .6 81 .8 96 .6 96 .6 78 .8 96 .9 77 .8 96 .9 79 .6 96 .9 96 .7 sp or ot hr ix s p. 1 55 r j 85 r j 15 2r j 19 0a r j m m c c it s1 /n l 4 b t2 a/ b t2 b it s1 /n l 4 b t2 a/ b t2 b it s1 /n l 4 it s1 /n l 4 jq 29 28 21 jq 29 28 32 jq 29 28 28 jq 29 28 20 s. in fla ta d e h oo g s. in fla ta s. in fla ta s. in fla ta s. in fla ta s. in fla ta jn 61 81 88 .1 a y 49 54 37 .1 jn 61 81 88 .1 a m 49 83 45 .1 jn 61 81 88 .1 jn 61 81 88 .1 93 .3 88 .1 93 .2 86 .2 96 .5 93 .6 a j – j ar os zo w ie c, c – c hr oś ni ca , m – m ie le c ophiostomatoid fungi 205 results identification of ophiostomatoid species. morphological investigation showed that seven ophiostomatoid species were collected, namely leptographium piriforme, ophiostoma canum, o. floccosum, o. minus, grosmannia cucullata-like and two unknown fungi, ophiostoma sp. 1 and sporothrix sp. 1. strain classified as grosmannia cucullata-like produced in culture dark brown perithecia with straight or curved neck ended ostiolar hyphae. ascospores were hyaline, onecelled, lunate, with a thick hyaline sheath. this fungus formed pesotum anamorph, stipe branched at the apex , with cylindrical conidia (figs 2-5). sporothrix sp. 1 (fig. 6) was characterized by producing micronematous conidiogenous cells arising orthotropically from undifferentiated hyphae. conidia were produced directly on denticles, clavate; no ascomata were observed. strains referred to as ophiostoma sp. 1 presented both pesotum and sporothrix synanamorphs in culture (figs 7-10) while no ascomata were observed. conidiophores were synnematous, branched at the apex, single but often also in loosely arranged groups. conidiogenous cells of sporothrix state were micronematous, mononematous, hyaline, arising orthoor slightly plagiotropically from undifferentiated aerial hyphae. conidia were produced directly on denticles, clavate, slightly curved. comparison of its sequences obtained for isolates in this study, with sequences from genbank, indicate that our isolates could also be assigned to seven taxa. dna sequences of the majority of species identified here were identical with reference sequences, confirming their identity (tab. 1). its rdna sequences of g. cucullata-like isolate showed 99% similarity with g. cucullata strains from genbank. in the its tree, ophiostoma sp. 1 grouped most closely to ophiostoma quercus in o. piceae complex (fig. 11). similarity search using a ß-tubulin showed highest relatedness with ophiostoma pseudotsugae (<81.8%) and ophiostoma breviusculum (78.8%) (tab. 1). based on sequence data of the its regions and ß-tubulin, the unknown sporothrix sp. collected by us was most closely related to s. inflata and s. schenckii in s. schenckii-o. stenoceras complex (tab. 1, fig. 11). isolation frequency. all the species were found at very low frequency (tab. 2). in culture, 43 % (28 isolates) of a total of 64 fungal isolates obtained from pine shoots table 2 number of isolates and frequency (in parentheses)* of fungi species associated with pine shoots infested by tomicus spp. fungal species sites total mielec chrośnica jaroszowiec babimost grosmannia cucullata-like 1(1) 1(0.3) leptographium piriforme 1(1) 2(2) 3(0.8) ophiostoma canum 1(1) 3(1) 4(1.0) ophiostoma floccosum 2(2) 2(0.5) ophiostoma minus 2(2) 10(5) 3(1) 15(2.0) ophiostoma sp. 1 17(6) 11(2) 28(2.0) sporothrix sp. 1 1(1) 6(3) 4(4) 11(2.0) total no. isolates 2 29 15 18 64 total no. shoots and shoot fragments (in parentheses) 100 (600) 100 (600) 100(600) 100(600) 400 (2400) *frequency = (no. of shoots from which a particular fungus was isolated/total number of shoots) x 100 206 r. jankowiak and m. kolařík infested by tomicus spp. in poland, represented ophiostoma sp. 1. this species was isolated from shoots with an average isolation frequency of 2% (from 0% at jaroszowiec to 6% at chrośnica). the second most frequent species (15 isolates) was o. minus, with frequencies ranging from 0% at mielec to 5% at jaroszowiec. the third taxon, sporothrix sp. 1 (11 isolates) occurred at an average frequency of 2% (from 0% jaroszowiec to 4% babimost). leptographium piriforme, o. canum, g. cucullatalike strain and o. floccosum were isolated from shoots infested by tomicus spp. with very low frequencies (tab. 2). fig. 11. phylogram obtained from neighbour-joining analyses of dna sequences of nuclear its region. bootstrap support values (1000 replicates) above 50% are indicated at the nodes. isolates with markers represent those collected and sequenced in this study. ophiostomatoid fungi 207 discussion this is the first survey of ophiostomatoid species associated with pine shoots damaged by adult individuals of tomicus spp. yielded in 64 strains isolated from four pine forests stands in poland. based on morphological characteristics and dna sequence comparison, four species of the genus ophiostoma s.str. and after one grosmannia, leptographium and sporothrix were confirmed as associated with fallen pine shoots. three species, namely o. canum, o. floccosum and o. minus, have previously been recorded in europe in association with tomicus spp. beetles (kirisits 2004). however, o. floccosum is reported here for the first time from poland. this fungus is widely distributed in europe primarily on pine trees, but usually it is recorded at low frequency (linnakoski et al. 2010). two species, ophiostoma sp. 1 and sporothrix sp. 1 probably represent new taxa. according to molecular data, isolates of ophiostoma sp. 1 obtained in this study resided in the o. piceae-complex including species having allantoid ascospores and both pesotum and sporothrix synanamorphs. ophiostoma sp. 1 seems to be closely related to o. quercus. the morphological characteristics of sporothrix sp. 1 broadly resemble species of the s. schenckii–o. stenoceras complex. sequence data and phylogenetic analyses also placed sporothrix sp. 1 in the s. schenckii–o. stenoceras complex. sequences of sporothrix sp. 1 even formed a separate lineage within s. schenckii–o. stenoceras complex. only single isolate represented grosmannia cucullata-like, thus its taxonomical status remains unclear and requires confirmation based on greater number of isolates. based on its sequences this strain is closely related to isolates of g. cucullata. this species was also morphologically similar to g. cucullata but had shorter perithecial neck and larger ascospores. grosmannia cucullata has been reported from picea abies (l.) karst., p. sylvestris and larix decidua mill. in association with various species of bark beetles (kirisits 2004) and cerambycids (jankowiak, kolařik 2010a). the taxonomic position of ophiostoma sp. 1, g. cucullata-like and sporothrix sp. 1 will be further discussed after a more critical study has been concluded. we presumed that new t. piniperda and t. minor individuals may efficiently introduce spores of ophiostomatoid fungi to young pine shoots during maturationfeeding. unexpectedly, the frequencies of the ophiostomatoid species were very low. none species was consistently isolated from each of the studied pine stands. unfavorable growth conditions in very fresh tissues of shoots could be the reason the lower than expected frequency of these fungi. our study demonstrates that among fungal associates of tomicus spp. in poland only o. minus may infest fresh pine shoots during maturation-feeding. these results confirmed the findings of jankowiak and kurek (2006), who isolated o. minus from t. piniperda galleries at relatively high frequencies in 10 weeks after the main beetles attack. the ability of o. minus to colonize the shoots results from its ability to tolerate low concentration of oxygen (solheim et al. 2001). this physiological feature agreed with the high level of virulence of this species to pine (lieutier et al. 1989; solheim, långström 1991; solheim et al. 1993; solheim et al. 2001; jankowiak et al. 2007). however, low and very variable frequency of o. minus indicates that its role in damaging of pine shoots during the shoot feeding phase is rather limited. 208 r. jankowiak and m. kolařík our studies showed that shoots laying on the forest floor may be relatively often colonized by ophiostomatoid fungi that are not closely associated with tomicus spp. beetles, primarily by ophiostoma sp. 1 and sporothrix sp. 1. ecology of these fungi is however unknown. we presume that various antropods as spiders, ants and dipterans may vectored propagules of ophiostomatoid species. in nature, we often observed unidentified spiders in the pitch of shoots damaged by tomicus spp. similar observations were made by greif et al. (2006), who isolated l. piriforme from various arthropods taxa. the presence of l. piriforme in pine shoots demonstrated in our study, could be the result of such vectoring as well. interestingly, this fungus has been described by greif et al. (2006) from various arthropods collected in an aspen-dominated forest in western canada and until 2010 it has been known only from this country. our study showes that l. piriforme occurs pine forest habitat in europe and may suggest that the species can develop on a wider range of host plants. its pathogenicity has not been fully recognized yet. according to jankowiak and kolařik (2010b), l. piriforme appears to be a weak pathogen of pinus sylvestris. sporothrix species placed in the o. stenoceras-s. schenckii complex are reported to live mostly as saprotrophs on wood or as soil fungi (de beer et al. 2003). six species that had been isolated directly from soil but one of the recent studies (de meyer et al. 2008) have revealed next three new species associated with wood and soil: sporothrix stylites de meyer, z.w. de beer & m.j. wingf., s. humicola de meyer, z.w. de beer & m.j. wingf. and s. lignivora de meyer, z.w. de beer & m.j. wingf. we suspect that sporothrix sp. 1 may have a similar ecological niche as above mentioned species because this fungus was isolated from pine shoots laying often directly on the sandy soil. conclusions the aim of this study was to characterize ophiostomatoid species associated with pine shoots damaged by tomicus spp. in poland. the diversity of reported was low, with seven species found. in addition, all the species were recorded at low frequencies. our research show that the pine shoots damaged by tomicus spp. beetles are poorly colonized by fungal associates of these insects. only ophiostoma minus, the most important associate of t. piniperda in poland, was relatively often isolated from fallen pine shoots. two other frequent species, ophiostoma sp. 1 and sporothrix sp. 1 probably represent undescribed taxa. this is also the first report of ophiostoma floccosum from poland. acknowledgement. we thank wilhelm de beer (department of microbiology and plant pathology, forestry and agricultural biotechnology institute (fabi), university of pretoria, pretoria, south africa) for his assistance with species identification. ophiostomatoid fungi 209 references borkowski a. 2001. threats to pine stands by the pine shoot beetles tomicus piniperda (l.) and t. minor (hart.) around a sawmill in southern poland. j. appl. entomol. 125: 489–492. de beer z.w., harrington t.c., vismer h.f., wingfield b.d., wingfield m.j. 2003. phylogeny of the ophiostoma stenoceras–sporothrix schenckii complex. mycologia 95 (3): 434–441. de meyer e.m., de beer z.w., summerbell r.c., moharram a.m., de hoog g.s., vismer h.f., wingfield m.j. 2008. taxonomy and phylogeny of new woodand soil-inhabiting sporothrix species in the ophiostoma stenoceras-sporothrix schenckii complex. mycologia 100 (4): 647–661. francke-grosmann h. 1952. über die ambrosiazucht der beiden kiefernborkenkäfer myelophilus minor htg. und ips acuminatus gyll. meddn. st. skogforsk. inst. 41: 1–52. gibbs j.n., inman a. 1991. the pine shoot beetle tomicus piniperda as a vector of blue-stain fungi to windblown pine. forestry 64: 239–249. glass n.l., donaldson g.c. 1995. development of primer sets designed for use with the pcr to amplify conserved genes from filamentous ascomycetes. appl. environ. microbiol. 61: 1323–1330. greif m.d., gibas c.f., currah r.s. 2006. leptographium piriforme sp. nov., from a taxonomically diverse collection of arthropods collected in an aspen-dominated forest in western canada. mycologia 98: 771–780. jankowiak r. 2006. fungi associated with tomicus piniperda in poland and assessment of their virulence using scots pine seedlings. ann. for. sci. 63: 801–808. jankowiak r. 2008. fungi associated with tomicus minor on pinus sylvetris in poland and their succession into the sapwood of beetle-infested windblown trees. can. j. for. res. 10: 2579–2588. jankowiak r., kurek m. 2006. the early stages of fungal succession in scots pine phloem and sapwood infested by the pine shoot beetle – tomicus piniperda. dendrobiology 56: 27–36. jankowiak r., bilański p. 2007. fungi associated with tomicus piniperda l. in an area close to a timber yard in poland. j. appl. entomol. 131 (8): 579–584. jankowiak r., kolařík m. 2010a. diversity and pathogenicity of ophiostomatoid fungi associated with tetropium species (coleoptera: cerambycidae) colonizing picea abies in poland. folia microbiol. 55 (2): 145–154. jankowiak r., kolařík m. 2010b. leptographium piriforme – first record for europe and potential pathogenicity. biologia 65 (4): 754–757. jankowiak r., rossa r., bilański p. 2007. contribution to pathogenicity of three blue-stain fungi associated with the pine sawyer beetle monochamus galloprovincialis (coleoptera: cerambycidae) to scots pine in poland. phytopathol. pol. 46: 37–46. kirisits t. 2004. fungal associates of european bark beetles with special emphasis on the ophiostomatoid fungi. 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taxonomy, ecology and pathogenicity. american phytopathological society, st paul, mn. grzyby z rodzaju ophiostoma s.l. wyizolowane z opadłej cetyny sosnowej w polsce streszczenie grzyby należące do grupy gatunków w przeszłości zaliczanych do rodzaju ophiostoma s.l. są znane ze współżycia z owadami z rodzaju tomicus żerującymi na sośnie zwyczajnej pinus sylvestris l. jednakże bardzo mało wiemy o grzybach występujących w pędach sosny, uszkodzonych przez te owady. celem badań było poznanie składu gatunkowego i frekwencji występowania grzybów z tej grupy związanych z pędami sosny zwyczajnej uszkodzonymi przez żer uzupełniający cetyńców. badania przeprowadzono w latach 2007-2008 w czterech drzewostanach sosnowych zlokalizowanych w różnych częściach polski. grzyby były identyfikowane na podstawie cech morfologicznych, a wybrane, reprezentatywne szczepy grzybów poddano także amplifikacji i sekwencjonowaniu fragmentów its1, 5.8s, its2 oraz fragmentów genu ß-tubuliny. ogółem, z pędów uszkodzonych przez cetyńce, otrzymano 64 izolaty grzybów reprezentujące siedem gatunków grzybów należących do ophiostoma s.l. dwa z nich, ophiostoma sp. 1 i sporothrix sp. 1 prawdopodobnie reprezentują taksony nowe dla nauki. niepewny pozostaje również status taksonomiczny izolatu zaklasyfikowanego jako grosmannia cucullata-like. wszystkie stwierdzone gatunki charakteryzowały się niską częstością występowania. wśród nich, ophiostoma minus, ophiostoma sp. 1 i sporothrix sp. 1 były izolowane z 2.0% pędów. pozostałe gatunki grzybów (leptographium piriforme, ophiostoma canum, o. floccosum i grosmannia cucullata-like) zasiedlały pędy sosny sporadycznie. wśród zidentyfikowanych grzybów o. floccosum, ophiostoma sp. 1 i sporothrix sp. 1 zostały stwierdzone w polsce po raz pierwszy. 2014-01-01t23:49:26+0100 polish botanical society ambispora gerdemannii and glomus badium, two species of arbuscular fungi (glomeromycota) new for europe and poland, respectively 1janusz błaszkowski, 1beata czerniawska, 1sławomir kowalczyk, 2katarzyna turnau and 3szymon zubek 1department of plant protection, west pomeranian university of technology słowackiego 17, pl-71-434 szczecin, janusz.blaszkowski@zut.edu.pl 2department of ecological microbiology, institute of environmental sciences jagiellonian university, gronostajowa 7, pl-30-387 kraków, katarzyna.turnau@uj.edu.pl 3laboratory of mycology, institute of botany, jagiellonian university lubicz 46, pl-31-512 kraków, szymon.zubek@uj.edu.pl błaszkowski j., czerniawska b., kowalczyk s., turnau k., zubek s.: ambispora gerdemannii and glomus badium, two species of arbuscular fungi (glomeromycota) new for europe and poland, respectively. acta mycol. 45 (1): 17–25, 2010. morphological characters of spores, as well as sporocarps and spores of ambispora gerdemannii and glomus badium, respectively, arbuscular fungi of the phylum glomeromycota, are described and illustrated. additionally, the known distribution of these species in both poland and the other regions of the world is presented. ambispora gerdemannii was not earlier reported from europe, and g. badium is a new fungus for poland. key words: glomeromycota, occurrence, distribution introduction analysis of collections of arbuscular-mycorrhizal fungi made over the last 27 years in poland and other regions of the world included two unrecorded species, i. e., ambispora gerdemannii (s.l. rose, b.a. daniels et trappe) spain, oehl et sieverd. and glomus badium oehl, redecker et sieverd. the former fungus has not so far been reported from europe, and the latter species has not been reported to occur in poland to date. the aims of this paper are to describe and illustrate these species and present their distribution in both poland and the world. acta mycologica vol. 45 (1): 17–25 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 18 j. błaszkowski et al. materials and methods the methods of collection of soil samples, establishment of trap and one-species cultures, growth conditions, isolation and preparation of spores and the determination of properties of their subcellular structure, as well as the terminology of spore structure and colours, the nomenclature of plants and fungi, and the authors of the fungal names used are as those presented previously (błaszkowski, czerniawska 2005). microphotographs were recorded on a sony 3cdd color video camera coupled to an olympus bx 50 compound microscope equipped with nomarski differential interference contrast optics. voucher specimens were mounted in polyvinyl alcohol/lactic acid/glycerol (pvlg; omar, bollan and heather 1979) and a mixture of pvlg and melzer’s reagent (1:1, v/v) on slides and deposited in the department of plant pathology (dpp). colour microphotographs of spores of am. gerdemannii and g. badium can be viewed at the url http://www.agro.ar.szczecin.pl/~jblaszkowski/. descriptions of the species ambispora gerdemannii (s.l. rose, b.a. daniels et trappe) c. walker, vestberg et schuessler spores formed singly in the soil; arise blastically at the tip of a hyphal branch (pedicel) on the neck of a sporiferous saccule (figs 1 and 8). spores pale yellow (4a3) to maize yellow (4a6), globose to subglobose, (150-)210(-250) μm diam, sometimes ovoid, 150-190 × 210-250 μm. subcellular structure of spores consists of a spore wall and two inner germination walls (figs 3-6 and 8). spore wall composed of three layers (layers 1-3). layer 1, forming the spore surface, evanescent, pale yellow (4a3) to maize yellow (4a6), (1.7-)3.3(-5.6) μm thick when intact, usually with deep fissures due to sloughing with age (figs 1 and 2). layer 2 semi-flexible, smooth, hyaline (1.2-)2.5(-3.7) μm thick, continuous with the wall of the pedicel (figs 3-6 and 8). layer 3 flexible to semi-flexible, hyaline, 0.5-1.0 μm thick, usually tightly adherent to the lower surface of layer 2 and, hence, difficult to see (figs 3-5 and 8). germination wall 1 comprises two fragile, smooth, hyaline layers (layers 1 and 2), (0.5-)1.1(-1.6) μm and (0.6-)1.4(-2.1) μm thick, respectively, usually tightly adherent to one another and readily cracking to form line slits or polygonal pieces in crushed spores (figs 3-8). germination wall 2 consists of three smooth, hyaline layers (layers 1-3; figs 3-5 and 8). layer 1 flexible to semi-flexible, 0.5-0.8 μm thick, always tightly adherent to layer 2 and, thereby, very difficult to observe (figs 3-5). layer 2 semi-flexible, finely laminate, hyaline, (1.7-)2.8(-4.1) μm thick (figs 3-5 and 8). layer 3 flexible to semiflexible, 0.5-0.8 μm thick (figs 3-5), rarely separating from layer 2. in melzer’s reagent, only spore wall layer 1 stains orange (6a6) to pastel red (8c5; fig. 5). pedicel 10-15 μm long, 15-27 μm wide at the spore base, positioned 80-120 μm from the base of the sporiferous saccule, consisting of a hyaline wall continuous with the wall of the saccule neck, the spore wall, and layer 1 of germination wall 1 (fig. 8). spori ferous saccule hyaline to yellowish white (4a2), globose to subglobose, (185-)200(-275) μm figs 1-8. ambispora gerdemannii. 1. spore (sp) associated with the neck (n) of sporiferous saccule (ss). 2. partly deteriorated spore wall layer 1 (swl1). 3-5. spore wall layers 1-3 (swl1-3), layers 1 and 2 of germination wall 1 (gw1l1+2), and layers 1-3 of germination wall 2 (gw2l1-3). 6. spore wall layers 1-3 (swl1-3) and layers 1 and 2 of germination wall 1 (gw1l1+2) with line splits in slightly crushed spore. 7. disintegrated layers 1 and 2 of germination wall 1 (gw1l1+2) in vigorously crushed spore. 8. spore wall layers 1-3 (swl1-3), layers 1 and 2 of germination wall 1 (gw1l1+2), layers 1-3 of germination wall 2 (gw2l1-3), and pedicel (p) with a wall continuous with spore wall layer 2. fig. 1, spore in lactic acid. figs 2-4 and 6-8, spores crushed in pvlg. fig. 5, spore crushed in pvlg+melzer’s reagent. fig. 1, bright field microscopy; figs 2-8, differential interference contrast. scale bars for fig. 1=50 μm, for figs 2-8=10 μm. figs 9-16. glomus badium. 9. sporocarp. 10. hyphal plexus (hp). 11. interspore mycelium (im). 12. cystidium-like structure (cs). 13. spore wall layers 1-3 (swl1-3). 14. spore wall layers 1-3 and interspore mycelium (im); septum (s) of the subtending hypha formed by spore wall layer 3 is indicated. 15. subtending hyphal wall layers 1 and 2 (shwl-2). 16. plug (p) in the lumen of subtending hypha. figs 9 and 11-16, spores crushed in pvlg. fig. 10, spore crushed in pvlg+melzer’s reagent. figs 9-16, differential interference contrast. scale bars for fig. 9=20 μm, for figs 10-16=10 μm. ambispora gerdemannii and glomus badium 19 diam, occasionally ovoid, 160-200 × 210-260 μm, formed at the end of a funnelshaped neck (fig. 1). wall of sporiferous saccule semi-flexible, semi-permanent, hyaline to yellowish white (4a2), 3.5-8.8 μm thick, smooth in young specimens, roughened because of a patchy sloughing of its outer surface, composed of two to three tightly adherent layers, usually difficult to observe. saccule neck hyaline to yellowish white (4a2), 200-250 μm long, 30-45 μm wide at the base of the saccule, 20-25 μm wide at the spore base, then gradually tapering up to 8-10 μm wide. collections examined. poland. przelewice (53o06’n, 15o05’e), under thuja occidentalis l., 29 sept. 1989, błaszkowski j. 1722 and 1783-1801 (dpp); truskaw (52°18’n, 20°46’e), from the root zone of juniperus communis l., 9 aug. 1990, błaszkowski j. 1802-1804 (dpp); trzciel (52°22’n, 15°52’e), among roots of equisetum arvense l., 13 aug. 2004, błaszkowski j. 1805-1806 (dpp); romanówek (52°17’n, 15°22’e), from under achillea millefolium l. and hypericum perforatum l., 13 aug. 2004, błaszk. j., s.n. (dpp); lubrza (52°18’n, 15°27’e), under h. perforatum, 13 aug. 2004, błaszk. j. s.n. (dpp); tatra mountains, among roots of campanula polymorpha witasek, cardaminopsis neglecta (schul.) hayek, cerastium tatrae borbás, knautia kitaibelii (schult.) borbás, leucanthemum waldsteinii (sch. bip.) pouzar, melampyrum herbichii woł., sesleria tatrae (degen) deyl, and thymus sp., błaszkowski j. 2634-2643 (dpp). distribution and habitat. in poland, spores of ambispora gerdemannii were found in 19 samples of roots and rhizosphere soils of 14 species of uncultivated plants. none of the almost 1500 root and soil mixtures coming from cultivated sites of poland contained spores of this fungus. the average abundance of am. gerdemannii spores in the samples examined was 4.7 and ranged from 1 to 29 in 100 g dry soil. the proportion of spores of this species in spore populations of all the arbuscular fungi recovered averaged 11.8% in a range of 2.1-66.7%. the average abundance of species of arbuscular fungi in samples in which spores of am. gerdemannii occurred was 2.7 and ranged from 1 to 6 in 100 g dry soil. the arbuscular fungi accompanying am. gerdemannii in the field were acaulospora bireticulata f.m. rothwell et trappe, a. capsicula błaszk., a. koskei błaszk., a. lacunosa j.b. morton, a. paulinae błaszk., ar. trappei (r.n. ames et linderman) j.b. morton et d. redecker, entrophospora infrequens (i.r. hall) r.n. ames et r.w. schneid., g. caledonium (nicol. et gerd.) trappe et gerd., g. claroideum n.c. schenck et s.m. sm., g. constrictum trappe, g. deserticola trappe, bloss et j.a. menge, g. ? etunicatum w.n. becker et gerd., g. fasciculatum (thaxt.) gerd. et trappe emend. c. walker et koske, g. geosporum (nicol. et gerd.) c. walker, g. macrocarpum tul. et c. tul., g. mosseae (nicol. et gerd.) gerd. et trappe, g. pansihalos s.m. berch et koske, an unrecognized glomus sp., pacispora scintillans (s.l. rose et trappe) sieverd. et oehl, scutellospora dipurpurescens j.b. morton et koske, and s. pellucida (nicol. et n.c. schenck) c. walker et f.e. sanders. although am. gerdemannii probably has a worldwide distribution, this species has been infrequently reported. most reports of am. gerdemannii come from the united states of america (allen, macmahon 1985; an et al. 1990, 1993; an, guo and hendrix 1993a; an, quo and hendrix 1993b; bever et al. 1996; koske, gemma and jackson 1977; nicolson, schenck 1979; rose, daniels and trappe 1979). additionally, spores of this fungus have been isolated in brazil (moreira-souza et al. 2003), colombia (dodd et al. 1990), and australia (morton, redecker 2001). in southern poland, turnau et al. (2001) recovered spores of a morphotype named glomus sp. hm-cl4 of molecular properties close (71%) to those of g. gerdemannii, the basionym of am. gerdemannii (walker et al. 2007). however, the authors did 20 j. błaszkowski et al. not show any morphological characters of the spores isolated and did not determine which of the two morphotypes of this fungus was found. mycorrhizal associations. spores of am. gerdemannii occurred among vesicular-arbuscular mycorrhizal roots of the plant species listed in the section “collection examined”. additionally, they were recovered from some trap cultures established from mixtures of roots and rhizosphere soils of these plants and with p. lanceolata as the plant host. unfortunately, many attempts to produce one-species cultures of am. gerdemannii failed. according to morton (2002) and morton & redecker (2001), mycorrhizae of am. gerdemannii consisted of only arbuscules and intraradical hyphae; no vesicles were found. all the structures were patchily distributed along roots and stained weakly or not at all in trypan blue. percentage mycorrhizal colonization always was very low, below 10%. notes. ambispora gerdemannii has originally been described as glomus gerdemanii s.l. rose, b.a. daniels et trappe from spores isolated from soils of cascade range and siskiyou mountains of oregon, usa (rose et al. 1979). spain et al. (2006) transferred g. gerdemannii to the genus appendicispora spain, oehl et sieverd. gen. nov., and walker et al. (2007) to the genus ambispora c. walker, vestberg et schuessler gen. nov. in the newly erected family ambisporaceae c. walker, vestberg et schuessler. the genus appendicispora has been excluded because a nomenclatural error (walker et al. 2007). the distinctive morphological properties of am. gerdemannii are the fragile bilayered inner wall 1 and the mode of differentiation of spores of this fungus. additionally, am. gerdemannii is a dimorphic fungus producing two morphotypes, acaulosporioid (as in acaulospora spp.) and glomoid (as in glomus spp.), a phenomenon rarely occurring in other species of arbuscular fungi. inner wall 1 of am. gerdemannii resembles a hardened glass. in slightly crushed spores, linear cracks appear on the surface of this wall. however, in spores vigorously crushed, this wall almost always breaks into small pieces. another characteristic of this wall is its glittering when seen in a polarized light. when observed under a dissecting microscope, am. gerdemannii spores may easily be confused with those of a. spinosa, a. thomii, am. appendicula and am. fennica. all the fungi form yellow brown and dull spores of a similar size range. spores of am. appendicula, am. fennica, and am. gerdemannii differ mainly in the properties of layers of inner wall 1. in the latter two species, both layers of this wall are smooth on both sides (spain et al. 2006; walker et al. 2007). in contrast, in the former fungus, the lower surface of layer 1 is ornamented with hemispherical protuberances, which impress hemispherical concave depressions on the upper surface of layer 2 during its differentiation (morton, redecker 2001; spain et al. 2006). additionally, in vigorously crushed spores, inner wall 1 of am. fennica and am. gerdemannii usually disintegrates, and that of am. appendicula remains its integrity. as concluded above (see comments on am. fennica), am. fennica and am. gerdemannii probably are congeneric. the subcellular structure readily separating am. gerdemannii from a. spinosa and a. thomii is the beaded outer layer and the flexible to plastic, dextrinoid inner layer present in the innermost wall of spores of the latter two species (vs. no such structures have been found in spores of ambispora spp.). additionally, in am. ambispora gerdemannii and glomus badium 21 gerdemannii the sporiferous saccule wall is continuous with spore wall layers 1–3 and layer 1 of inner wall 1, whereas in all known acaulospora spp. the sporiferous saccule wall is continuous with only spore wall layer 1. finally, only am. gerdemannii is a dimorphic fungus, forming both acaulosporioid and glomoid spores. the glomoid morphotype of am. gerdemannii has not so far been found in poland and the only report of its existence is that of morton and redecker (2001). glomus badium oehl, redecker et sieverd. spores occur in dense sporocarps in the soil (fig. 9) and on the surface of vesicular-arbuscular mycorrhizal roots. sporocarps brownish orange (6c8) to reddish brown (8e8), mainly ovoid to irregular, 200-500 × 290-680 μm (fig. 9), sometimes globose to subglobose, (250-)298(-320) μm diam, with 4-43 spores, radially originating from a hyphal plexus (fig. 10) and separated by an interspore mycelium (figs 11 and 14) and occasionally by cystidium-like structures (fig. 12). interspore mycelium consists of tightly interwoven, straight or branched, hyaline to pale orange (5a3) hyphae, (2.2-)5.8(-12.3) μm wide, with a wall (0.5-)1.1(-1.5) μm thick (figs 11 and 14). cystidium-like structures icicle-like, hyaline, 32.1-39.5 μm long, 9.6-12.0 μm wide at the base, with a wall (1.8-)2.0(-2.3) μm thick (fig. 12). both the interspore mycelium and the cystidium-like structures branch out from the hyphal plexus. inside sporocarps frequently with sand grains and soil debris. spores formed blastically at the tip of hyphae developing from a hyphal plexus positioned in the centre of sporocarps (fig. 10). hyphal plexus composed of thickwalled, 6.9-10.3 μm thick, two-layered hyphae: a hyaline, evanescent, 0.5-0.8 μm thick outer layer, usually highly deteriorated and difficult to see in mature spores and pale orange (5a3) to brownish orange (6c8), permanent, (1.2-)2.3(-3.4) μm thick inner layer (fig. 12). spores brownish orange (6c8) to reddish brown (8e8), usually ovoid, 35-55 × 60-70 μm, rarely globose to subglobose, (35-)55(-65) μm, usually with one subtending hypha (figs 9, 10 and 14-16), occasionally with two. subcellular spore structure consists of a spore wall comprising three layers (layers 1-3; figs 13 and 14). layer 1 evanescent, hyaline to pale orange (5a3), (0.7-)2.0(-3.4) μm thick, frequently highly or completely sloughed in mature spores (figs 13 and 14). layer 2 laminate, brownish orange (6c8) to reddish brown (8e8), (3.7-)4.5(-6.9) μm thick (figs 13 and 14). layer 3 flexible, pale yellow (4a3) to light brown (6d8), (0.5)1.7(-2.0) μm thick, usually tightly adherent to the lower surface of layer 2 (figs 13 and 14). subtending hypha brownish orange (6c8) to reddish brown (8e8), straight or recurved, cylindrical or slightly flared, rarely slightly constricted, (8.1-)9.5(-10.0) μm wide at the spore base (figs 10, 15 and 16). wall of subtending hypha brownish orange (6c8) to reddish brown (8e8), (2.9-)4.4(-7.4) μm thick at the spore base, continuous with spore wall layers 1 and 2 (fig. 15). pore 1.0-1.7 μm wide at the spore base, occluded by spore wall layer 3 inserted in the lumen of the subtending hypha up to 14.0 μm below the spore base (fig. 14) or/and by thickening of spore wall layer 2 and wall layer 2 of subtending hypha, in which a hyphal plug occasionally forms (fig. 16). collections examined. poland. szczecin (53°26’n, 14°35’e), under glycine max (l.) merr., 5 sept. 1985, błaszkowski j. 2644-2646 (dpp); przelewice, among roots of malus × purpurea rehder, 16 july 1985, błaszkowski j. 2647-2649 (dpp); pomierzyn (53°20’n, 15°51’e), in the roots zone of lupinus luteus l., 9 aug. 1985, błaszkowski j. 2650-2652 (dpp); szczecin, in the rhizosphere soil of anthriscus sylvestris (l.) hoffm., 18 oct. 1985, błaszkowski j. 2653 (dpp); kołbacz (53o18’n, 14o49’e), under trifolium pratense 22 j. błaszkowski et al. l., 4 aug. 1985, błaszkowski j. 2654-2655 (dpp); szczecin, from under allium schoenoprasum l., 3 sept. 1985, błaszk. j. s.n. (dpp); międzyzdroje (53°56’n, 14°26’e), among roots of avena sativa l., 14 july 1985, błaszk. j. s.n. (dpp); przelewice, in the roots zone of acer palamatum thunb., 15 may 1985, błaszk. j. s.n. (dpp); żelistrzewo (54°41’n, 18°27’e), under triticum aestivum l., 24 aug. 1985, błaszk. j. s.n. (dpp); hel (54o36’n, 18o49’e), in the rhizosphere soil of thuja occidentalis, 21 aug. 1985, błaszk. j. s.n. (dpp); jarosławki (53°32’n, 15°01’e), under zea mays l., 25 july 1985, błaszk. j. s.n. (dpp); żelistrzewo, among roots of ficaria verna huds., 3 april 1989, błaszk. j. s.n. (dpp); hel, in the root zone of t. occidentalis, 28 sept. 1988, błaszk. j. s.n. (dpp); szczecin, under t. occidentalis, 18 oct. 1985, błaszk. j. s.n. (dpp); żelistrzewo, near roots of festuca ovina l., 25 june 1986, błaszk. j. s.n. (dpp); jastrzębia góra (54o50’n, 18o18’e), under crataegus monogyna jacq., 30 oct. 1987, błaszk. j. s.n. (dpp); żelistrzewo, from under rubus idaeus l., 22 aug. 1987, błaszk. j. s.n. (dpp); płoty (53°48’n, 15°16’e), among roots of agrostis stolonifera l., 22 sept. 1987, błaszk. j. s.n. (dpp); jastrzębia góra, in the rhizosphere soil of j. communis l., błaszk. j. s.n. (dpp); szczecin, under heracleum sphondylium l., 22 oct. 1987, błaszk. j. s.n. (dpp); osłonino (54°42’n, 18°28’e), in the root zone of helictotrichon pubescens (huds.) pilg., 22 june 1988, błaszk. j. s.n. (dpp); chałupy, among roots of festuca arundinacea schreb., 28 sept. 1988, błaszk. j. s.n. (dpp); kuźnica (54°41’n, 18°34’e), associated with roots of ammophila arenaria (l.) link, 28 sept. 1988, błaszk. j. s.n. (dpp); władysławowo (54°47’n, 18°26’e), on roots of rosa rugosa thunb., 7 july 1989, błaszk. j. s.n. (dpp); bolesław (50°00’n, 18°12’e), under an unrecognized grass, 17 sept. 1989, błaszk. j. s.n. (dpp); nowy ujków (50°18’n, 19°26’e), among roots of agropyron repens (l.) p. beauv., 17 sept. 1989, błaszk. j. s.n. (dpp); leszna góra (49°42’n, 18°43’e), associated with roots of c. monogyna, 8 sept. 1989, błaszk. j. s.n. (dpp); pieski (52°25’n, 15°26’e), under hordeum vulgare l., 12 aug. 2004, błaszk. j. s.n. (dpp); połupin (52°03’n, 15°06’e), from under rumex acetosella l., 12 aug. 2004, błaszk. j. s.n. (dpp); jemiołów (52°21’n, 15°16’e), in the root zone of cirsium arvense (l.) scop., 13 aug. 2004, błaszk. j. s.n. (dpp); trzciel (52°22’n, 15°52’e), among roots of corynephorus canescens (l.) p. beauv., 15 aug. 2004, błaszk. j. s.n. (dpp); łagów (51°59’n, 15°16’e), under silene latifolia poir. ssp. alba (mill.) greuter et burdet, 13 aug. 2004, błaszk. j. s.n. (dpp); jemiołów, among roots of h. perforatum, 13 aug. 2004, błaszk. j. s.n. (dpp); zarzyń (52°23’n, 15°25’e), from under e. arvense, 13 aug. 2004, błaszk. j. s.n. (dpp); trzciel, in the rhizosphere soil of juncus effusus l., 15 aug. 2004, błaszk. j. s.n. (dpp). distribution and habitat. in poland, g. badium was recovered from 39 fieldcollected mixtures of roots and rhizosphere soils. of them, 15 came from under nine species of cultivated plants, and the others represented 20 species of wild plants. the spore abundance of g. badium was much higher among roots of wild plants (av. 126.3; range 1-850 in 100 g dry soil) than cultivated plants (av. 5.2; range 1-20 in 100 g dry soil). the proportion of spores of this fungus in spore populations of all arbuscular fungi isolated also was higher in the root zone of wild plants (av. 45.2%; range 0.9-100%) than cultivated ones (av. 10.8%; range 0.9-40.7%). the species abundance in the root and soil samples taken from under cultivated plants averaged 5.0 with a range of 4-7 in 100 g dry soil, and that under wild plants averaged 3.7 and ranged from 1 to 9 in 100 g dry soil. the arbuscular fungi co-occurring with g. badium were a. polonica błaszk., a. capsicula, a. cavernata błaszk., a. gedanensis błaszk., a. lacunosa, a. mellea spain et n.c. schenck, a. paulinae, an unrecognized acaulospora sp., g. aggregatum n.c. schenck et s.m. sm. emend. koske, g. caledonium, g. constrictum, g. deserticola, g. ? etunicatum, g. fasciculatum, g. fuegianum, g. geosporum, g. microcarpum tul. et c. tul., g. macrocarpum, g. mosseae, unrecognized glomus spp., paraglomus occultum (c. walker) j.b. morton et d. redecker, scutellospora armeniaca błaszk., and scutellospora dipurpurescens. apart from poland, g. badium has been found among roots of grasses and grassland plants growing in soils of ph 6-8 and located in germany, france, switzerland, and italy (oehl et al. 2005). ambispora gerdemannii and glomus badium 23 mycorrhizal associations. the sporocarps of g. badium found by the authors of this paper were formed among and occasionally on vesicular-arbuscular mycorrhizal roots of the plant species listed in the section “collection examined”. in trap cultures with root and rhizosphere soil mixtures of the plant species sampled, g. badium rarely produced spores. all one-species cultures of this fungus established were unsuccessful. oehl et al. (2005) also failed to obtain g. badium in one-species cultures and, hence, properties of mycorrhizae of this fungus remain unknown. however, phylogenetic molecular analyses showed g. badium to be a member of glomus group a (oehl et al. 2005) sensu schwarzott, walker and schüßler (2001), comprising fungi known to form vesicular-arbuscular mycorrhizae (błaszkowski 2003). the most distinguishing characters of g. badium are its small sporocarps lacking a peridium and composed of many, brownish orange to reddish brown, relatively small spores (fig. 9). the innermost flexible to semi-flexible and coloured layer of the threelayered spore wall also is a diagnostic property of this species (figs 13 and 14). glomus badium probably forms spores only in multispored sporocarps, and single spores occasionally recovered from field-collected soil samples represented their fragments. examination of the ontogenesis of this species is needed to confirm this supposition. unfortunately, all attempts to produce one-species cultures of g. badium made by the authors of this paper and by oehl et al. (2005) failed. the interspore mycelium of the specimens of g. badium found by the authors of this paper morphologically corresponds to that characterized by oehl et al. (2005). however, the original description of this fungus does not inform of the cystidiumlike structures occurring between spores of the polish sporocarps of this fungus. the function of the cystidia-like structures in sporocarps of g. badium probably is similar to that of cystida of agaricales, in which they appear to serve as spacers between basidia (ulloa, hanlin 2000). of the three wall layers of spores of g. badium, the two outer ones are typical of most species of the genus glomus, in which the outermost layer sloughs with age and adheres to a laminate structural layer (figs 13 and 14). spore wall layer 3 of g. badium is thin, pale yellow (4a3) to light brown (6d8), and usually tightly adheres to the lower surface of the brownish orange (6c8) to reddish brown (8e8) layer 2 in even vigorously crushed spores (figs 13 and 14). hence, it is difficult to see, especially in young and freshly matured spores. in older spores, this layer usually is slightly thicker and more frequently separates from the middle laminate layer. additionally, this layer is most visible at the spore base where it forms a straight or curved septum of the subtending hypha continuous with its part adherent to the laminate spore wall layer 2 (fig. 14). apart from g. badium, species of the genus glomus forming small and compact sporocarps with small, brownish orange to reddish brown spores are g. cuneatum mcgee et cooper, g. fuegianum (speg.) trappe et gerd., g. invermaium i.r. hall, and g. rubiforme (gerd. et trappe) r.t. almeida et n.c. schenck. compared with g. badium, sporocarps of g. cuneatum are much larger (2-12 mm vs. 200-500 × 290680 μm in g. badium) and have a peridium (fig. 9; vs. no peridium in g. badium; mcgee and trappe 2002). additionally, the spore wall of the latter species consists of only one laminate layer lightening from black to hyaline towards their inside, whereas the spore wall of the former fungus comprises three layers, of which the laminate layer is darkand uniform-coloured (figs 10, 12 and 16). 24 j. błaszkowski et al. according to thaxter (1922), g. fuegianum spores coming from spegazzini’s original collection were reddish brown as are those of g. badium. in contrast, all g. fuegianum spores found by one of the authors (j. b.) of this paper in poland, those loaned from kew, united kingdom (błaszkowski, madej and tadych 1998), and those revealed in australia by mcgee and trappe (2002) were pale yellow to yellow brown. moreover, the spores were frequently surrounded by branched and convoluted hyphae (błaszkowski 2003) that never occur on g. badium spores (błaszkowski, pers. observ.; oehl et al. 2005). finally, the radial arrangement of spores in sporocarps of g. fuegianum is regular, and irregular in g. badium sporocarps. glomus badium differs from g. invermaium in the formation of smaller (200-500 × 290-680 μm vs. up to 1 mm across) and more compact sporocarps with regularly distributed spores (fig. 9; vs. loose sporocarps with randomly distributed spores in g. invermaium; błaszkowski, pers. observ.; hall 1977). other differences are the number and the phenotypic properties of the spore wall layers of these fungi. the spore wall of g. badium comprises three layers, of which the outermost one sloughs with age (figs 13 and 14). in contrast, the outermost spore wall layer of g. invermaium spores is persistent and the spore wall of this species lacks spore wall layer 3 of g. badium. the main difference between g. badium and g. rubiforme resides in the wall structure of their spores. the spore wall of g. rubiforme comprises only two layers similar in colour and phenotypic properties to layers 1 and 2 of the spore wall of g. badium (błaszkowski 2003; gerdemann, trappe 1974). acknowledgment. this study was supported in part by the committee of scientific researches, a grant no. 2 po4c 041 28 and 164/n-cost/2008/0. references allen m. f., macmahon j. a. 1985. impact of disturbance on cold desert fungi: comparative microscale 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with two new genera archaeospora and paraglomus, based on concordant molecular and morphological characters. mycologia 93: 181–195. nicolson t. h., schenck n. c. 1979. endogonaceous mycorrhizal endophytes in florida. mycologia 71: 178–198. oehl f., redecker d., sieverding e. 2005. glomus badium, a new sporocarpic mycorrhizal fungal species from european grasslands with higher soil ph. j. appl. bot. and food qual. 79: 38–43. omar m. b., bollan l., heather w. a. 1979. a permanent mounting medium for fungi. bull. brit. mycol. soc. 13: 31–32. rose s., daniels b. a., trappe j. m. 1979. glomus gerdemannii sp. nov. mycotaxon 8: 297–301. schwarzott d., walker c., schüßler a. 2001. glomus, the largest genus of the arbuscular mycorrhizal fungi (glomales) is nonmonophyletic. mol. phylogen. evol. 21: 190–197. spain j. l., sieverding e., oehl f. 2006. appendicispora: a new genus in the arbuscular mycorrhiza-forming glomeromycetes, with a discussion of the genus archaeospora. mycotaxon 97: 163–182. thaxter r. 1922. a revision of the endogonaceae. proc. am. acad. arts and sci. 57: 291–351. turnau k., ryszka p., gianinazzi-pearson v., tuinen d. van 2001. identification of arbuscular mycorrhizal fungi in soils and roots of plants colonizing zinc wastes in southern poland. mycorrhiza 10: 169–174. ulloa m., hanlin r. t. 2000. illustrated dictionary of mycology. aps press. the american phytopathological society. st. paul, minnesota. walker c., vestberg m., demircik f., stockinger h., saito m., sawaki h., nishmura i., schüßler a. 2007. molecular phylogeny and new taxa in the archaeosporales (glomeromycota): ambispora fennica gen. sp. nov., ambisporaceae fam. nov., and emendation of archaeospora and archaeosporaceae. mycol. res. 111: 137–153. ambispora gerdemannii and glomus badium, dwa gatunki grzybów arbuskularnych (glomeromycota) nowe odpowiednio dla europy i polski streszczenie opisano i zilustrowano cechy morfologiczne zarodników oraz sporokarpów i zarodników odpowiednio ambispora gerdemannii i glomus badium, grzybów arbuskularnych z gromady glomeromycota. ponadto przedstawiono poznane rozmieszczenie tych gatunków zarówno w polsce, jak i innych regionach świata. ambispora gerdemannii nie była dotychczas notowana w europie, a g. badium jest nowym grzybem dla polski. 2014-01-01t11:50:07+0100 polish botanical society european record of subramaniula thielavioides on opium poppy martin pastirčák1 and katarína pastirčáková2 1slovak agricultural research centre, research institute of plant production bratislavská cesta 122, sk-92168 piešťany, uefemapa@hotmail.com 2slovak academy of sciences, institute of forest ecology, branch of woody plants biology akademická 2, sk-94901 nitra, uefezima@hotmail.com pastirčák m., pastirčáková k.: european record of subramaniula thielavioides on opium poppy. acta mycol. 44 (1): 7–9, 2009. in the course of a study of fungal biodiversity of opium poppy (papaver somniferum) plants collected in production area of slovakia, an ascomycete belonging to the genus subramaniula was isolated. the fungus identified as subramaniula thielavioides has been reported for the first time from slovakia. this record also represents the first european locality. brief morphological description of the fungus based on an isolate from flower petals of opium poppy is provided. key words: subramaniula thielavioides, sordariales, papaver somniferum, slovakia introduction the genus subramaniula arx (chaetomiaceae, sordariales) includes two species, subramaniula irregularis p.f. cannon & d. hawksw. and s. thielavioides (arx, mukerji & n. singh) arx designated as the type species. morphologically, subramaniula thielavioides differs from s. irregularis by much smaller ascospores and irregularly shaped but usually lacrimiform or obovate ascospores. according to arx (1985), morphological features of subramaniula thielavioides show affinities to some species of thielavia zopf, chaetomium kunze and podospora ces. the genus subramaniula differs from these genera by urniform, pale ascomata with a very wide apical ostiole surrounded by a ring of hyaline cells. arx et al. (1978) observed non-ostiolate ascomata occasionally. an anamorphic connection is unknown. only cannon (1986) observed a few hyphae with chains of considerably swollen cells, producing thinwalled globose or ellipsoidal bodies, probably related to the anamorph. during survey of fungal mycobiota of opium poppy (papaver somniferum l.) the ascomycete s. thielavioides was isolated. subramaniula thielavioides is not included acta mycologica vol. 44 (1): 7–9 2009 8 m. pastirčák and k. pastirčáková in the checklist of fungi of slovakia (lizoň, bacigálová 1998). there are no published data on the biology and natural occurrence of this ascomycete in slovakia and no species of subramaniula have been recorded in the country so far. the species s. thielavioides is newly described for slovakia. material and methods flower petals of opium poppy were collected at locality dvory nad žitavou (western slovakia) in june 2006. fresh material was surface-sterilized by immersion in 80% alcohol for 1 min and 5% commercial bleach solution of sodium hypochlorite savo (bochemie, bohumín, czech republic) for 1 min and plated on sna agar (leslie, summerell 2006) in 90 mm petri dishes. petri plates were incubated at 22°c. the morphological features of the fungus sporulating in pure culture were examined by means of standard light microscopy (jenamed2, carl zeiss jena, germany). slide preparations were stained with lactophenol blue solution (merck, darmstadt, germany). the morphological structures (ascocarps, asci and ascospores) were pho-(ascocarps, asci and ascospores) were photographically documented by digital camera olympus camedia c-4000 zoom. representative material has been deposited in the mycological herbarium of research institute of plant production in piešťany, slovakia. taxonomic description subramaniula thielavioides (arx, mukerji & n. singh) arx, proc. indian acad. sci., pl. sci. 94 (2-3): 344 (1985). basionym: achaetomium thielavioides arx, mukerji & n. singh, persoonia 10 (1): 144 (1978). the following description is based on a fresh isolate from flower petals of opium poppy. colonies on sna agar with hyaline submerged mycelium; aerial mycelium scanty and hyaline; ascomata (perithecia) started appearing on the 10th day of incubation, scattered over or below (into) the agar surface. ascomata globose, 160–460 μm in diam., without short neck, the collar of thin-walled hyaline cells around the ostiole. ascomatal hairs absent; narrow hyaline hyphae emanating from various points on the ascoma surface. periphyses not seen. asci clavate, short-stalked, very thin-walled, without apical structures, evanescent at a very early stage, 8-spored, 75-96 × 10.5-18 μm. ascospores widely fusiform, brownish, relatively thick-walled, 20.5-34 × 11.5-19 μm, 1-celled, with a conspicuous subapical germ pore up to 3.5 μm wide (fig. 1). anamorph not seen. specimen examined. slovakia, dvory nad žitavou, opium poppy field, 47.9937 n, 18.2678 e, living flower petals of papaver somniferum, 23 jun 2006, leg. m. pastirčák. record of subramaniula thielavioides 9 biometric data of ascomata and asci containing ascospores in general correspond to those given by arx et al. (1978), arx (1985), and cannon (1986). the measurements of ascomata, asci and ascospores of the slovak specimen compared with those recorded by previous authors are given in table 1. on the basis of morphological description and measurements of reproductive structures, the fungus was identified as subramaniula thielavioides. table 1 subramaniula thielavioides: measurements of ascomata, asci and ascospores compared with previous descriptions source of data ascomata (in diam., μm) asci size (μm) ascospores size (μm) present study (average ± sd) 330 ± 92 85 ± 7.5 × 13 ± 2.5 26 ± 3 × 14.5 ± 1.5 range (min-max) 160-460 75-96 × 10.5-18 20.5-34 × 11.5-19 arx et al. (1978) 150-240 50-78 × 18-35 21-27 × 13-16 arx (1985) 130-200 – 23-27 × 13-17 cannon (1986) 130-200 × 110-160 45-69 × 18-31 22-26 × 11.5-14.5 according to cannon (1986), subramaniula thielavioides is almost certainly no pathogen. although arx et al. (1978) includes this species in coprophilous fungi; the fungus was isolated from human nails, vertebrate dung, and soil. until recently, distribution of this species was known from india (arx et al. 1978; arx 1985; cannon 1986; arx et al. 1988). lately, s. thielavioides was isolated from biological soil crusts at the mashash farm in the negev desert, israel (grishkan et al. 2006). online database of the cabi bioscience uk centre herbarium (imi) includes record of subramaniula sp. isolated from fruit of/associated with capsicum from pakistan in 1998 (imi 379385, not seen). the finding of s. thielavioides on flower petals of papaver somniferum from slovakia represents the first record of the species in the country and the first known locality in europe. acknowledgements. this study was supported by the ministry of agriculture of slovak republic, grant no. 2006 uo 27/091 05 01/091 05 11 and partially supported by science and technology assistance agency under the contract no. apvt-27-009904. references arx j. a. von 1985. on achaetomium and a new genus subramaniula (ascomycota). proc. indian acad. sci. (plant sci.) 94 (2–3): 341–345. arx j. a. von, figueras m. j., guarro j. 1988. sordariaceous ascomycetes without ascospore ejaculation. beih. nova hedwigia 94: 1–104. arx j. a. von, mukerji k. g., singh n. 1978. a new coprophilous ascomycete from india. persoonia 10 (1): 144–146. cannon p. f. 1986. a revision of achaetomium, achaetomiella and subramaniula, and some similar species of chaetomium. trans. br. mycol. soc. 87 (1): 45–76. grishkan i., zaady e., nevo e. 2006. soil crust microfungi along a southward rainfall gradient in desert ecosystems. eur. j. soil biol. 42 (1): 33–42. leslie j.f., summerell b.a. 2006. the fusarium laboratory manual. blackwell publishing, iowa, usa. lizoň p., bacigálová k. 1998. fungi. (in:) k. marhold, f. hindák (eds). checklist of non-vascular and vascular plants of slovakia. 1 ed., veda, bratislava: 101–227. fig. 1. subramaniula thielavioides: a – globose ascoma, scale bar: 100 μm; b – asci with ascospores, scale bar: 20 μm; c – ascospores, scale bar: 15 μm. 2014-01-01t11:48:31+0100 polish botanical society notes on caloplaca lucifuga (teloschistales, ascomycota) in poland dariusz kubiak1 and anna zalewska2 1 department of mycology, warmia and mazury university in olsztyn oczapowskiego 1a, pl-10-957 olsztyn, darkub@uwm.edu.pl 2 department of botany and nature protection, warmia and mazury university in olsztyn plac łódzki 1, pl-10-727 olsztyn, annazalw@uwm.edu.pl kubiak d., zalewska a.: notes on caloplaca lucifuga (teloschistales, ascomycota) in poland. acta mycol. 44 (2): 239–248, 2009. the current knowledge on the occurrence of caloplaca lucifuga, a rare lichen with an inconspicuous crustose sorediate thallus, is discussed. both previous and new localities are presented. the most important data on the ecology and general distribution of the species are given. diagnostic characters related to the morphology, anatomy and chemistry of c. lucifuga that help to differentiate it from similar species are described. key words: lichenized fungi, sorediate species, caloplaca, distribution, n poland introduction caloplaca lucifuga belongs to a group of crustose, sterile sorediate lichens that are relatively poorly known in poland (nowak, tobolewski 1975; śliwa, tønsberg 1995; śliwa 1996; kukwa 2005a, b, 2006; kukwa, szymczyk 2006; kukwa, kubiak 2007). despite its small-sized thallus, this very rare species has distinct, characteristic morphological and chemical features that help to distinguish it from other similar taxa. its special ecological requirements also make the species noteworthy. c. lucifuga is associated with the bark of old deciduous trees, especially oaks. it seems to have potential indicator properties that may be used to identify high-biodiversity sites and forest communities of special value for protection purposes (cf. rose 1992; arup 1997; nordén et al. 2007). caloplaca lucifuga was first reported in poland by lipnicki (1993) from the bory tucholskie forest. its localities were later noted in the puszcza borecka forest by zalewska (2000) and in the puszcza białowieska forest by sparrius (2003). specimens of c. lucifuga were collected from the puszcza białowieska forest by h. sipman and p. van den boom as early as in 1988; however, the exact location of the acta mycologica vol. 44 (2): 239–248 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 240 d. kubiak and a. zalewska sites is unknown (unpbl. data acc. to sparrius 2003; sparrius, pers. comm. 2009). data on the occurrence of c. lucifuga in north-east poland are provided in a study by cieśliński (2003), who cites unpublished materials by j. nowak from the puszcza białowieska forest. the available scarce data on the distribution of caloplaca lucifuga in poland are given in this study and its new localities recorded in the pojezierze olsztyńskie lakeland are presented on the map. habitat requirements of c. lucifuga identified so far suggest that further localities of the species are likely to occur in poland. the aim of the study was to present the species and to indicate the most important diagnostic characters that help to find it in field studies and to recognise it in herbarium materials. material and methods the material was collected in 1995–1997 and 2005–2008. specimens were identified with standard morphological and anatomical methods as well as using spot colour reaction tests to differentiate secondary metabolites (purvis et al 1992; orange et al. 2001). the square number of the modified atpol grid (cieśliński, fałtynowicz 1993) was given for all localities. the nomenclature of lichens follows fałtynowicz (2003) and diederich et al. (2009). the material was deposited in the herbarium of the department of mycology, warmia and mazury university in olsztyn (oltc-l) and in the herbarium of the department of botany and nature protection, warmia and mazury university in olsztyn (ols-l). results caloplaca lucifuga thor, lichenologist 20 (2): 175 (1988). morphology. thallus endophloedic, inconspicuous or visible as a thin grey film with scattered small blisters. photobiont chlorococcoid 7–14 µm diam., algal cells in clusters (in the thallus section). soralia numerous, 0.15–0.35 mm in diam., forming by a break-up of blisters, pale yellow, yellowish grey to dirty yellow-orange-brown, delimited, but sometimes lying very close to one another, rounded or flexuose, flat, ulcerose or slightly elevated, when erumpent through the bark (fig. 1). soredia granular or farinose 17–24 µm diam. apothecia not seen so far, including the polish material. chemistry. according to thor (1988), the species produces hydroxyanthraquinone derivatives: parietin (major) and fallacinal (minor). spot test reactions: thallus c–, k–, pd–; soralia c–, k+ violet-red, pd–. ecology. caloplaca lucifuga was recorded in old oak-lime-hornbeam and oakhornbeam forests in poland, at sun-exposed or moderately shaded sites with a relatively high air humidity. the species occurred on the bark of old oaks and hornbeams notes on caloplaca lucifuga 241 (usually with a trunk girth of over 300 cm) growing by mid-forest roads, less frequently inside forest communities. the species was observed on the bark of quercus robur together with acrocordia gemmata, arthonia byssacea, calicium adspersum, c. salicinum, c. viride, chrysothrix candelaris, cliostomum corrugatum, lecanora expallens, lepraria incana, l. vouauxii, lobaria pulmonaria, ochrolechia turneri, pertusaria amara, phlyctis argena and ramalina sp. in the pojezierze olsztyńskie lakeland. the group of accompanying taxa in the puszcza borecka forest included species such as buellia griseovirens, chaenotheca trichialis, pertusaria albescens, p. coccodes and p. hemisphaerica on oaks, as well as arthonia didyma, a. ruana, bacidia subincompta, bacidina assulata, lecanora intumescens, l. glabrata, opegrapha varia, o. viridis, pyrenula laevigata and p. nitida on the bark of hornbeams. according to literature sources, caloplaca lucifuga clearly prefers the bark of old deciduous trees in woodland tree stands described in different parts of europe as „wooded pastures” or „wooded meadows” (thor 1988). such ecosystems consist of more or less scattered, aged trees and a mosaic of grazing or, less frequently, mowing meadows (kirby et al. 1995). known mostly from scandinavia and great britain, they also occur in, for instance, estonia. these sites have for a long time functioned as extensive pastures and are characterised by a specific ecological continuity. they are important habitats of many rare lichens everywhere (thor 1988; laundon 1992; lepik, jüriado 2008). c. lucifuga is an indicator species of such areas (arup 1997; coppins, coppins 2002). c. lucifuga also finds similar conditions in very old parks (thor 1988; laundon 1992; hallingbäck 1995). futhermore, it occurs in denser forests, although usually on the bark of trees growing on the forest edge, e.g. by forest roads. caloplaca lucifuga usually colonises deeply cracked bark of very old oaks (quercus robur, q. petraea) with a breast diameter of 1–2 m. it was observed on the bark of castanea, fagus, tilia and ulmus considerably less frequently (thor 1988). the species is also known from the bark of aesculus hippocastanum (brunialti et al. 2001; aptroot et al. 2001), acer pseudoplatanus (palice et al. 2003), carpinus betulus (lipnicki, pers. comm. 2009; zalewska 2000), castanea sativa (poelt 1994; lopez de silanez et al. 1999), fagus sylvatica (spier et al. 2008), tilia cordata (türk, wunder 1999) and ulmus glabra (berger, türk 1993). c. lucifuga was also recorded on dead wood (clerc 2004). thor (1988) stresses the species preference for high air-humidity areas (e.g. near lakes, the seaside). he also notes that the lichen mostly grows in deep bark cracks in well-lit sites while it also occurs outside the cracks on flat, raised surfaces, when the shading is greater. the majority of records of c. lucifuga from oak bark are reported for trees with natural properties of the periderm, not impregnated with dusts (thor 1988; thor, arvidsson 1999; ek, johannesson 2006). data from more diversified habitats (bratli, haugan 1997) suggest a slightly broader ecological amplitude of the species. based on studies conducted in the östergötland county in south-east sweden on sun-exposed precipices overgrown by oaks, ek et al. (1995) report that the breast diameter of 62% of old trees with fissured bark colonised by c. lucifuga ranged from 27 to 92 cm. therefore, the age of the tree and, to a lesser degree, its trunk diameter seem to be major factors conditioning the occurrence of c. lucifuga (thor 1988). 242 d. kubiak and a. zalewska caloplaca lucifuga grows in relatively rich epiphytic communities abundant in rare lichen species. of taxa mostly accompanying c. lucifuga on old oaks, arthonia byssacea, a. vinosa, buellia violaceofusca, calicium quercinum, chaenotheca hispidula, cliostomum corrugatum, cyphelium sessile, lecanographa amylacea, ramalina baltica incl. obtusata, schismatomma decolorans and s. pericleum, are particularly interesting (thor, arvidsson 1999; ek, johannesson 2006). according to wirth (1995), c. lucifuga is a component of communities of the order chrysothrichetalia candelaris wirth, represented by, e.g., numerous „calicioid lichens”. world distribution. caloplaca lucifuga occurs mostly in europe, where it has a broad distribution range. the species in known from denmark, france, germany, sweden, great britain and italy (thor 1988) as well as from luxembourg (diederich 1989), belgium (diederich et al. 1991), estonia (ekman et al. 1991), austria (berger, türk 1993), poland (lipnicki 1993), portugal (van den boom and giralt 1996), norway (bratli, haugan 1997), finland (vitikainen et al. 1997), spain (etayo 1991), latvia (sundin, thor 1990), the czech republic (palice et al. 2003), lithuania (motiejūnaitė et al. 2003), switzerland (clerc 2004), slovakia (palice et al. 2006), bulgaria (spier et al. 2008) and croatia (partl 2009). it seems that the occurrence range of c. lucifuga in europe is related to the distribution of deciduous forests of the temperate zone. localities of the species in sweden and norway in the scandinavian peninsula are situated only in its southern part belonging to the nemoral zone. the lichen also occurs in the sub-mediterranean zone in the south of europe, probably at localities situated mostly in the mountains (cf. lópez de silanes et al. 1999; brunialti et al. 2001). outside europe, c. lucifuga is known only from south america. two specimens of the lichen were collected by h. sipman and j. aguirre in columbia (lichen herbarium berlin, bgbm – www). caloplaca lucifuga is usually known from few localities in individual countries. it is locally very frequent only in southern sweden (hultengren 1994; ek et al. 1995), from where thor and arvidsson (1999) report ca 200 localities. on the other hand, ek and johannesson (2006) report as many as 400 records (58% of all records in sweden) only from the östergötland county in south-east sweden. it is also a fairly frequent lichen in the vosges in france (aptroot et al. 2001). remarks. as the number of its localities is small, c. lucifuga is considered to be vulnerable in many countries. it is classified as critically endangered (cr) in finland (rassi et al. 2001) and is believed to be endangered (en) in austria (türk, haffelner 1999), denmark (søchting, alstrup 2002), switzerland (scheiddeger et al. 2002) and the czech republic (liška et al. 2008). it is classified as vulnerable (vu) in great britain (woods, coppins 2003) and norway (timdal et al. 2006). it is thought to be near threatened (nt) in sweden (gärdenfors 2005) and estonia (randlane et al. 2008). the species has not been red-listed in poland due to very scarce data (cieśliński et al. 2006). however, as habitats characteristic of c. lucifuga are becoming extinct and the number of its localities is small, the taxon seems to fully deserve to be included in the next edition of the red list. distribution in poland. c. lucifiga is known from ten localities in northern and north-east poland. it occurs in the bory tucholskie forest (lipnicki 1993), pusz-puszcza borecka forest (zalewska 2000), puszcza białowieska forest (sparrius 2003; cieśliński 2003) and in the pojezierze olsztyńskie lakeland (present work) (fig.2). notes on caloplaca lucifuga 243 localities known prior to this paper. atpol grid square bc 58 – bory tucholskie forest, vicinity of the ‘krzywe koło w pętli wdy’ reserve (lipnicki 1993). bf 03 and bf 13 – pojezierze ełckie lakeland, puszcza borecka forest (zalewska 2000; cieśliński 2003). cg 55 – równina bielska plain: puszcza białowieska forest (cieśliński 2003, locality cited as pers. comm. by j. nowak). cg 66 – puszcza białowieska forest, białowieża, białowieża forest inspectorate, near the road bordering on forest sections nos 452b and 453a (sparrius 2003; l. sparrius, pers. comm. 2009). specimens examined. be 62 – pojezierze olsztyńskie lakeland: las warmiński nature reserve, forest section no 705d, 53°38’59.7”n, 20°30’03.1”e, 8 july 2008, leg. d. kubiak (oltc-l 3042). be 63 – ‘las warmiński’ nature reserve, forest section no 214l, 53°46’49.0”n, 20°25’30.3”e, 1 may 2005, leg. d. kubiak (oltc-l 1864). be 83 – pojezierze olsztyńskie lakeland, ‘dęby napiwodzkie’ nature reserve, 30 july 2008, leg. d. kubiak (oltc-l 3227). bf 03 – pojezierze ełckie lakeland, puszcza borecka forest, borki forest inspectorate, diabla góra forest district, local square 38, 54°08’n, 22°05’e, at the edge of the oak-lime-hornbeam forest by the ‘dębowa linia’ forest road, 18 july1996, leg. a. zalewska (ols-l); local square 40, 54°08’n, 22°09’e, 13 june 1996, leg. a. zalewska (ols-l), rev. b. j. coppins, 1998. bf 13 – puszcza borecka forest, borki forest inspectorate, lipowo forest district, ‘borki’ nature reserve, forest section no 19a, local square 48, 54°07’n, 22°06’e, 18 july 1997, leg. a. zalewska (ols-l). bf 14 – puszcza borecka forest, czerwony dwór forest inspectorate, rogonie forest district, local square 61, 54°06’n, 22°11’e, 13 july 1995, leg. a. zalewska (ols-l). fig. 2. distribution of caloplaca lucifuga in poland (in the atpol square grid): 1 – known records; 2 – new records. 244 d. kubiak and a. zalewska discussion although it forms diminutive and inconspicuous thalli, caloplaca lucifuga is a fairly characteristic species. it is distinguished by a pale-grey, very thin endophloedic thallus and delimited soralia, yellow-grey to dirty yellowish-orange-brown reacting with k+ violet-red. caloplaca chrysophthalma degel. bears the greatest resemblance to c. lucifuga. the former differs from caloplaca lucifuga by having a thicker, epiphloedic thallus, grey to yellow (thor 1988), yellowish-green, yellow-green or grey (wetmore 2004), with distinct and delimited, usually bright orange-yellow soralia. it rarely produces immersed, poorly developed thalli with scattered, light yellow soralia (laundon 1992; wetmore 2004). furthermore, c. chrysophthalma produces bright orange apothecia, not always present (thor 1988; laundon 1992; wetmore 2004). clusters of algal cells, present in c. lucifuga, are absent on cross section (thor 1988). both species have slightly different chemical properties. c. chrysophthalma also contains parietin (major) as well as additional substances: emodin (minor) and fragilin (minor), instead of fallacinal. soralia and apothecia react with k+ violet-red. the distribution of both taxa also differs. c. chrysophthalma is mostly known from north america (wetmore 2004) and from northern europe, where it occurs in great britain (laundon 1992), norway (ihlen 1997) and denmark, estonia and sweden (thor, arvidsson 1999). it has few localities in other european countries as well as in mongolia (cf. sources cited by culberson et al. 2008) and china (wetmore 2004). this species has been not recorded in poland so far (cf. fałtynowicz 2003). moreover, caloplaca chrysophthlma differs from c. lucifuga by its ecological preferences. it is usually associated with trees having natural eutrophic or enriched, dusted bark. in europe, it is known from such phorophytes as acer, fraxinus, tilia, ulmus or less frequently from quercus growing in roadside alleys or other open sites (hallingbäck 1995; thor, arvidsson 1999). in north america, it is a forest epiphyte often found on, old trees of populus tremuloides (wetmore 2004). tlc analysis is not helpful in species identification when fine thalli, such as the ones used in this study, with a very low content of accessory substances, are examined (cf. thor 1988). thallus thickness and colour as well as the colour of soralia remain major characters differentiating c. lucifuga from c. chrysophthalma. habitat data which should, however, be interpreted with care (cf. bratli, haugan 1997), may also be useful. it seems that some characters in the descriptions of the morphology and ecology (thor 1988; laundon 1992; wetmore 2004) of both representatives of the genus caloplaca slightly overlap. the lack of more extensive data available on the frequency of chemical substances recorded and their influence on the colour of soralia as well as data on the structure of fruitbodies and picnidia, not found in c. lucifuga so far, causes some confusion. more wide-ranging chemo-taxonomic examinations of very large collections belonging to 400 records from östergötland (ek, johannesson 2006) and from north america (405 collections, see wetmore 2004) could be helpful. an analysis of the genetic similarity of both taxa could also provide interesting results. buellia violaceofusca thor & muhr, another sorediate species, can be found together with caloplaca lucifuga on old oak trees growing in forests. the former has a similar, pale grey endophloedic thallus with slightly elevated soralia, often confluent notes on caloplaca lucifuga 245 and clearly darker, brown tinged violet. the species is known to be sterile (apothecia and picnidia have not been observed). all spot test reactions on the thallus and soralia of b. violaceofusca are negative (thor, muhr 1991). it is finally worth mentioning that the status of c. lucifuga is not clear since, similarly to many other sterile representatives, the species was placed in the genus caloplaca mostly based on the presence of antraquinones, parietin and fallacinal. these substances occur together with emodin also in other teloschistales belonging to chemosyndrome a (arup, grube 1999). based on molecular studies, søchting and lutzoni (2003) have recently shown that the morphological form of the thallus and the development type of its lower cortex are not good taxonomic characters differentiating the genera caloplaca, xanthoria and fulgensia. it was proven that, for instance, some species of the genus caloplaca with crustose thalli should be transferred to the genus xanthoria. conclusions based on the current knowledge on the identification and ecology of c. lucifuga and c. chrysophthalma as well as the distribution rate of characteristic habitats preferred by them, it may be suggested that caloplaca lucifuga is considerably more widespread in poland but has been poorly differentiated (cf. wirth 1997). detailed observations of old oak tree stands, especially within large forest complexes in ne poland, should be conducted. it seems possible that c. chrysophthalma could also be found in poland. acknowledgements. the authors are indebted to b. j. coppins (edinburgh) for the confirmation of identification of specimen from the puszcza borecka forest. we would like to express our sincere thanks to s. cieśliński (kielce), l. lipnicki (gorzów wlkp.) and l. sparrius (gouda) for the detailed data on localities they supplied. data were used for mapping. the anonymous reviewer is warmly thanked for valuable comments on the manuscript. references aptroot a., sparrius l., herk van k., bruyn de u. 2001. origin and distribution of recently described lichens from the netherlands. aktuelle lichenologische mitteilungen, nf 5: 13–25. http://www.almneu.de/nf5/alm-nf5.pdf arup u. 1997. skoglig kontinuitet. 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(in:) l. kappen (ed.). new species and novel aspects in lichenology and physiology of lichens. in honour of o. l. lange. biblioth. lichenol. 67: 277–289. woods r. g., coppins b. j. 2003. a conservation evaluation of british lichens. british lichen society, london. zalewska a. 2000. ekologia porostów puszczy boreckiej i jej obrzeży. doctoral thesis. university of warmia and mazury in olsztyn, 178 pp. and atlas, 302 pp. uwagi o występowaniu caloplaca lucifuga (teloschistales, ascomycota) w polsce streszczenie caloplaca lucifuga jest porostem znanym z europy oraz z dwóch niepublikowanych notowań z ameryki południowej (kolumbia). pomimo szerokiego zasięgu występowania na kontynencie europejskim, gatunek ten podawany jest w poszczególnych krajach zwykle z niewielu stanowisk. w polsce zarejestrowany został łącznie na dziesięciu stanowiskach w północnej i północno-wschodniej części kraju. w pracy przedstawiono dotychczas znane oraz nowe stanowiska tego gatunku stwierdzone w latach 2005–2008 na pojezierzu olsztyńskim. uzyskane wyniki oraz przegląd danych dotyczących ekologii tego gatunku wskazują, iż może być on bardziej rozpowszechniony w dużych kompleksach leśnych z udziałem starych drzew liściastych, zwłaszcza dębów. ze względu jednak na zanikanie siedlisk charakterystycznych dla c. lucifuga, gatunek ten zasługuje na umieszczenie go na krajowej „czerwonej liście” porostów zagrożonych wymarciem. porost ten ma potencjalne właściwości lichenoindykacyjne, dotychczas w polsce niewykorzystane, mogące mieć zastosowanie w wyznaczaniu miejsc o wysokiej bioróżnorodności oraz najcenniejszych zbiorowisk leśnych. epifit ten, pomimo drobnych rozmiarów ma bardzo charakterystyczne cechy morfologiczne i chemiczne pozwalające odróżnić go od innych podobnych taksonów. najbardziej istotne cechy diagnostyczne tego gatunku to jasnoszara, endofloedyczna plecha oraz liczne, zaokrąglone lub nieregularne, nie zlewające się ze sobą, jasnożółtawe, żółtawoszare do brudno żółto-pomarańczowo-brunatnych soralia, barwiące się od koh na karminowo-czerwono. fig. 1. habit of caloplaca lucifuga (oltc-l 3227); scale bar = 1 mm. 2014-01-01t11:49:42+0100 polish botanical society professor alina skirgiełło a lina skirgiełło was born on november 3, 1911 in the village klince in russia. she spent her early childhood with her parents jadwiga and mieczyslaw skirgiełło in novozybkov. after her father, a lawyer, had got employed in forestry, the family moved to posiołek bor (kostroma district) in the middle of forests of northern russia. the skirgiełło family came to poland after the world war i and stayed in białowieża, where alina attended primary school. a few years later they moved to grodno. after finishing her college and getting a secondary school certificate in 1931 alina skirgiełło decided to study biology in the warsaw university at the faculty of mathematics and natural sciences. she graduated and was granted her master of science degree in 1937. her master thesis, entitled “polskie naziemne grzyby rurkowe” (polish terrestrial tube fungi) on boletes and related genera, prepared under the supervision of professor bolesław hryniewiecki, was published as a part of the “planta polonica” in 1939, just before the second world war. alina skirgiełło started to work in the department of plant systematics and geography on the 1st january, 1939, as a researcher. during the war she still lived in warsaw. she was working as a simple gardener in the botanical garden, was active in the resistance movement and cooperated with the polish home army (ak). one of her great achievements was saving rare collections, herbaria and old books. she passed through dramatic events during the warsaw uprising in the summer 1944 when many of her colleagues, friends and relatives died, even the closest one – her brother olgierd. after the war, since 1945, alina skirgiełło has restarted her full time work in the department of plant systematics and geography and worked there till retiring in 1981. among her main interests were taxonomical problems in boletaceae, polyporaceae, russula, pluteus and other fungi, as well as distribution of mushrooms in poland and europe. in 1948 she was granted doctor’s degree for the thesis “genus russula in poland and neighbouring countries”. in the following years she published over 180 scientific articles and books. the most valuable are monographs of boletales and the genera russula, lactarius, volvariella and pluteus. all of them are fine illustrated by the author. they were published as parts of the series “flora polska – grzyby” (flora of poland – fungi), edited as a whole by professor skirgiełło. apart from mycology she dealt also with paleobotany in co-operation with hanna czeczott. she supervised at least 90 undergraduate and dozen graduate students. she provided opinions and reviews for numerous doctoral and habilitation theses, as well as in professor’s title evaluation procedures. professor alina skirgiełło, fully engaged as a scientist and teacher, found time to do huge organisational work. she became a head of the department of plant systematics and geography in 1960, then a dean of the faculty in the 5 years 1966-1975. for a couple of years she has been a director of the institute of botany, warsaw university. also, she worked effectively for popularisation of mycology. she organised exhibitions, courses, wrote and translated many articles and books for amateurs. there is no doubt that the mycological section of the polish botanical society, founded by her initiative in 1956, was one of her “beloved children”. up to the present, for 50 years, she has been its president and soul. in 1965 she established acta mycologica – the first polish mycological journal. professor skirgiełło was its editor-in-chief for 37 years; nowadays she is still its honorary editor. she received a host of awards and distinctions for her scientific and administrative achievements. she was honoured with several medals, e.g. of bologna university (1981), university of łódź (1989), university of warsaw (2001), władysław szafer scientific medal and polonia restituta knight’s cross. professor skirgiełło has been conferred honorary membership of the polish botanical society and the committee of botany (polish academy of sciences). on the 15th of september 2004, professor alina skirgiełło became the first honorary member of the european mycological association. in the laudation it was written: “professor alina skirgiełło is an exceptional founder member of the ema. she is the only person to have attended every congress of european mycologists. moreover, she has been not only a representative of poland; but she also, for many years, played a key role as the contact person in co-operation between mycologists of western and eastern europe. she always reported thoroughly the content of congress sessions, shared congress materials and information about the current trends in mycology with those mycologists who could not themselves attend congress for political or financial reasons. in september 1966 professor skirgiełło organised the iv cem in poland: 150 mycologists took part, half coming from the so called “socialist countries”, especially the soviet union. that congress was a milestone in integration of european mycologists. all members of the ema send her their congratulations and warm wishes”. on the occasion of her 95. birthday the editorial council of acta mycologica proposed to devote a volume of this journal to its founder – professor alina skirgiełło. as a result of spontaneous reaction of mycologists, over 60 papers have been received. to satisfy all contributors it has been decided that the submitted papers will be published in two volumes – 41 and 42 – both of which should be treated as equivalent parts of the jubilee edition. our most sincere thanks are due to all the authors who sent their articles to be included in jubilee volume of acta mycologica. we all would like to thank professor skirgiełło for everything she has done as mycologist, academic teacher and friend of us. we wish her all the best for next years! maria ławrynowicz and marta wrzosek łódź – warszawa 2006 6 professor alina skirgiełło 2014-01-01t11:42:53+0100 polish botanical society im memoriam professor władysław wojewoda 1932-2010 władysław wojewoda was born on may 20th, 1932, in the small town of przemyśl in se poland. after studies in the faculty of biology and earth sciences of the jagiellonian university in kraków he started working in the jagiellonian university botanical garden (1959) and after two years as a junior lecturer in the department of plant systematics and geography at the jagiellonian university (1961-1968). during that period he was involved in intensive mycological studies. his doctoral thesis was devoted to “macromycetes” of the ojców national park. władysław wojewoda worked also in the w. szafer institute of botany of the polish academy of science in the years 1969-2003. his interests focused on the taxonomy, ecology, chorology and conservation of macrofungi, especially auricularioid, tremelloid, dacrymecetoid, tulasnelloid and aphyllophoroid fungi. he is an author of monographs on auriculariales and tremellales in poland and europe (wojewoda 1977), red list of threatened macrofungi in poland (wojewoda, ławrynowicz 1986, 1992, 2006), macrofungi of north korea (wojewoda et al. 2004) and monumental checklist of the polish basidiomycetes (wojewoda 2003). not only are many articles and monographs (altogether ca. 300 publications) a great achievement of professor wojewoda but also a collection of fungi which he founded in w. szafer institute of botany. this collection includes ca. 53 000 specimens of fungi, and is one of the most important collections of fungi in poland. in 2000 the first fascicle of the atlas of the geographical distribution of fungi in poland edited by professor wojewoda was published. he was also a good teacher for students, doctoral students and amateurs of mycology. professor wojewoda carried out mycological research in babia góra, gorce, magura, ojców and tatra national parks. his studies in north korea and niepołomice forest near kraków deserve special attention. he also collaborated with mycologists from the czech republic (former czechoslovakia). professor w. wojewoda and professor b. gumińska wrote a basic key for identification of macromycetes for students and teachers. professor wojewoda was a recognized authority for mycologists in poland and europe and was a kind, good humored man. his last article on laricifomes officinalis was devoted to professor gumińska (wojewoda 2010). professor died on november 3, 2010, and was buried in the rakowicki cemetery in kraków. andrzej chlebicki 2014-01-01t11:52:09+0100 polish botanical society lawrynomyces, a new genus of corticioid fungi in the hymenochaetales dariusz karasiński department of mycology, w. szafer institute of botany polish academy of sciences lubicz 46, pl-31-512 kraków, d.karasinski@botany.pl karasiński d.: lawrynomyces, a new genus of corticioid fungi in the hymenochaetales. acta mycol. 48 (1): 5–11, 2013. the new genus lawrynomyces is proposed to accommodate hyphoderma capitatum, a predominantly european species growing on decayed coniferous wood. the genus belongs to the rickenella-clade in the order hymenochaetales, and it includes species characterized by resupinate basidiomes, monomitic hyphal system with simple septate hyphae, presence of cystidia and hyphidia, suburniform to subcylindrical, pedunculate basidia, and subglobose to broadly ellipsoid basidiospores with slightly thickened walls not staining in melzer’s reagent or cotton blue. published molecular data support the recognition of a distinct genus for hyphoderma capitatum. key words: taxonomy, new genus, hyphoderma introduction hyphoderma capitatum j. erikss. & å. strid was described from a specimen collected by kurt hjortstam in alingsås, sweden (eriksson, ryvarden 1975). among the members of the genus hyphoderma wallr., it was considered to be a discordant taxon due to its simple septate hyphae and slightly thickened wall of the basidiospores. recent phylogenetic studies revealed that hyphoderma is polyphyletic and that most species cluster in two distinct, monophyletic clades (larsson et al. 2004, 2006; larsson 2007a, b). the generic type hyphoderma setigerum (fr.) donk and related species were placed in the “phlebioid clade” (meruliaceae, polyporales). this hyphoderma s.str. lineage is differentiated from other genera of the “phlebioid clade” by presence of suburniform basidia, tubular cystidia and cylindrical or elliptical spores (larsson 2007a). the second lineage is in the rickenella-clade of the hymenochaetales. the members of this lineage produce clavate basidia and often unique cells called echinocysts and stephanocysts, which have a nutritional function (hallenberg 1990) and capture and infect nematodes (tzean, liou 1993). to accommodate the species from this lineage, larsson (2007a) reinstated the acta mycologica vol. 48 (1): 5–11 2013 doi: 10.5586/am.2013.001 6 d. karasiński genus peniophorella p. karst., typified by p. pubera (fr.) p. karst. systematic position of some species from both lineages were independently (intentionally or not) confirmed in other molecular studies (miettinen, larsson 2011; miettinen, rajchenberg 2012; nakasone, burdsall 2012; tellería et al. 2012). however, some species of hyphoderma, for example h. argillaceum (bres.) donk, h. capitatum, h. orphanellum (bourdot & galzin) donk and h. sibiricum (parmasto) j. erikss. & å. strid, were not included in hyphoderma or peniophorella in molecular phylogenetic studies. in all published phylogenies, hyphoderma capitatum was recognized as a member of the rickenella-clade in the hymenochaetales and clustered in a clade separate from species of peniophorella. morphologically, h. capitatum differs from peniophorella species in having hyphae without clamps, suburniform-pedunculate basidia, basidiospores with slightly thickened walls, and lacking echinocysts and stephanocysts. morphologic and molecular phylogenetic data suggest that hyphoderma capitatum should be accommodated in a new genus that is described and illustrated here. materials and methods examined specimens were collected from the kaszubski landscape park, northern poland, in 2007-2008. laboratory methods and abbreviations follow karasiński (2010). specimens are deposited in author’s reference fungarium (herb. d.k.) and duplicates are in kram. taxonomy lawrynomyces karasiński, gen. nov. mycobank mb 803871 etymology: the genus is named in honor of polish mycologist, professor maria ławrynowicz. basidiomata resupinate, effused, adnate, thin. hymenophore even, margin indeterminate, without rhizomorphs. hyphal system monomitic. hyphae with simple septa, thin to thick-walled, hyaline. cystidia present. hyphidia sometimes present. basidia suburniform to subcylindrical and constricted, more or less pedunculate, basally without clamps, with (2–)4 prominent sterigmata. basidiospores relatively large, broadly ellipsoid to subglobose, with slightly thickened walls and distinct apiculus, smooth, inamyloid, indextrinoid, acyanophilous. generic type: hyphoderma capitatum j. erikss. & å. strid. lawrynomyces capitatus (j. erikss. & å. strid) karasiński, comb. nov. figs 1-3 mycobank mb 803872 basionym: hyphoderma capitatum j. erikss. & å. strid, in eriksson & ryvarden, cortic. n. eur., coronicium-hyphoderma 3: 461 (1975). lawrynomyces, a new genus of corticioid fungi 7 basidiome resupinate, closely adnate, very thin, 60–150 μm thick (excluding projecting cystidia), forming linear or irregular patches, becoming confluent with age, up to 20 cm in the longest dimension, soft, membranaceous, initially white, then grayish-white to grayish-cream, finally gray with brownish tint, dirty ochraceousgray when dry. hymenophore smooth, discontinuous as seen by stereomicroscope, densely reticulate, pilose from projecting cystidia; margin indeterminate, sometimes abrupt, somewhat pruinose or minutely fimbriate to loosely reticulate in some parts. odor or taste not known. hyphal system monomitic, all hyphae with simple septa. subiculum absent or in well developed basidiomes as a very thin layer of basal hyphae, 2–4.5 μm wide, parallel to substrate, slightly thick-walled, hyaline, smooth, branching at right-angles; subhymenium 20–50(–70) μm thick, consisting vertically and loosely arranged hyphae, 2–4 μm wide, sparsely branched, somewhat sinuous, thin-walled or up to 0.2 μm thick, hyaline, without encrustation, but in mature specimens covered (in koh) with yellowish to brownish gray re sinous matter; hymenial layer composed of cystidia, basidioles, basidia and hyphidia. cystidia 55–105 μm long, with swollen base, 7–12(–16) μm, tapering towards the apex and usually capitate, apical bulb 7–9.5(–11) μm in diameter, projecting up to 60 μm above basidia, thin-walled or with slightly thickened walls except at apex, empty, not encrusted, apical bulb enclosed by hyaline or yellow matter observed in melzer’s reagent or cotton blue but dissolving in koh and other media; in some well developed specimens, short capitate cystidia produced from basal hyphae, approximately 30 μm long. basidia 32–48 × 10–13 μm, suburniform to subcylindrical and constricted, more or less distinctly pedunculate, with oily granules in the protoplasm, without basal clamp, with 4 prominent sterigmata ca. 10 × 2.5 μm, 2-sterigmate basidia not observed in studied material. hyphidia scarce, enclosed or only slightly projecting, obtuse at apex, 3–5 μm wide, hyaline, thin-walled, smooth. fig. 1. lawrynomyces capitatus – section through basidiomata. line drawing from coll. karasiński 970 (kram f-56623). scale bar = 10 μm. 8 d. karasiński fig. 2. lawrynomyces capitatus – young basidiomata on very rotted wood of pinus sylvestris stump. coll. karasiński 2744 (herb. d.k.). fig. 3. lawrynomyces capitatus – mature and well developed basidiomata on much rotted wood of picea abies stump. coll. karasiński 970 (kram f-56623?). lawrynomyces, a new genus of corticioid fungi 9 basidiospores (8.8–)9–13.2(–14) × (7–)8.5–11(–11.5) μm (lm=11.2 μm, wm=9.3 μm, q=1.1–1.4 μm, qm=1.2 μm, n=120/4), subglobose to broadly ellipsoid, hyaline, containing one large or many small oil drops, smooth, with thickened walls and prominent apiculus, inamyloid, acyanophilous. specimens examined: poland. pojezierze kaszubskie lakeland: kaszubski landscape park, lasy mirachowskie forests, ca. 1.5 km n of miechucino, managed mixed forest, on very rotted wood of picea abies (l.) h. karst. stump, 26 oct. 2007, leg. d. karasiński 970 (herb. d.k., kram f-56623); kaszubski landscape park, kurze grzędy reserve, on bark of picea abies stump, 4 dec. 2008, leg. d. karasiński 2703 (herb. d.k.); kaszubski landscape park, bącz, ca. 2.5 km sw of mirachowo, managed mixed forest, on very rotted wood of pinus sylvestris l. stump, 5 dec. 2008, leg. d. karasiński 2725 (herb. d.k., kram f-56624); the same locality, substrate and date, leg. d. karasiński 2744 (herb. d.k.). distribution and ecology: lawrynomyces capitatus is an european taxon, reported from sweden (eriksson, ryvarden 1975), norway (ryvarden et al. 2003), finland (kotiranta et al. 2009), denmark (hjortstam 1984), germany (ostrow, dämmrich 2010), switzerland (breitenbach, kränzlin 1986), belgium, croatia, czech republic, estonia and france (bernicchia, gorjón 2010). it was reported from southern poland (wojewoda 2003) and from three new localities from northern poland reported here. in addition to europe, it was reported from turkey in asia minor (sesli, denchev 2011). it is not known from the tropics, subtropics and the southern hemisphere (hjortstam, ryvarden 2007). lawrynomyces capitatus is associated with much decayed wood of conifers. in one case, a basidiome was collected from the bark of picea abies stump in an early stage of decomposition. discussion the main morphological features of the genus lawrynomyces are a monomitic hyphal system with clampless hyphae, suburniform and more or less pedunculate basidia with prominent sterigmata, and smooth, hyaline, inamyloid, acyanophilous, broadly ellipsoid basidiospores with slightly thickened walls. the basidia of lawrynomyces are similar to those observed in species of hyphoderma s.str. but are typically pedunculate, i.e. with a stalk. the phylogenetic position of hyphoderma capitatum supports recognition of separate genus for this species. the other species that may be a member of the genus lawrynomyces is hyphoderma orphanellum (bourdot & galzin) donk. although h. orphanellum has basidia and cystidia of similar shape to lawrynomyces, it has clamped hyphae, thin-walled spores, encrusted hyphidia, and a dense, agglutinated subiculum (eriksson, ryvarden 1975; bernicchia, gorjón 2010). in some molecular phylogenetic studies h. orphanellum is shown to be in the same, well supported monophyletic clade with lawrynomyces capitatus (larsson 2007a; miettinen, larsson 2011; nakasone, burdsall 2012). however, these studies used the same its and/ or lsu sequence of h. orphanellum derived from nh 12208 from russia. another 10 d. karasiński sequence of h. orphanellum was included in the phylogenetic tree presented by tellería et al. (2012). in this study, h. orphanellum was placed in the peniophorella clade. because of these conflicting results, i accept a narrow concept of the genus which so far is monotypic. acknowledgements. marcin piątek, anna ronikier (kraków, poland) and an anonymous reviewer are thanked for valuable comments and suggestions to improve this manuscript. this work was partly supported by the polish ministry of science and higher education (grant no. n n304 079535 for the years 2008-2011) and statutory funds of w. szafer institute of botany pas. references bernicchia a., gorjón s.p. 2010. corticiaceae s.l. fungi europaei 12: 1-1008. breitenbach j., kränzlin f. 1986 fungi of switzerland 2. heterobasidiomycetes (jelly fungi), aphyllophorales (non-gilled fungi), gasteromycetes (puffballs). verlag mykologia, lucerne. eriksson j., ryvarden l. 1975. the corticiaceae of north europe 3: 284-546. hallenberg n. 1990. ultrastructure of stephanocysts and basidiospores in hyphoderma praetermissum. mycol. res. 94: 1090-1095. http://dx.doi.org/10.1016%2fs0953-7562%2809%2981339-7 hjortstam k. 1984. corticiaceous fungi of northern europe – check-list of the species in the nordic countries. windahlia 14: 1-30. hjortstam k., ryvarden l. 2007. checklist of corticioid fungi (basidiomycotina) from the tropics, subtropics and the southern hemisphere. synopsis fungorum 22: 27-146. karasiński d. 2010. polish resupinate russulales: the genus vararia. acta mycol. 45 (1): 45-56. kotiranta h., saarenokasa r., kytövuori i. 2009. aphyllophoroid fungi of finland. a checklist with ecology, distribution and threat categories. norrlinia 19: 1-223. larsson k.-h. 2007a. molecular phylogeny of hyphoderma and the reinstatement of peniophorella. mycol. res. 111: 186-195. http://dx.doi.org/10.1016%2fj.mycres.2006.10.002 larsson k.-h. 2007b. re-thinking the classification of corticioid fungi. mycol. res. 111: 1040-1063. http:// dx.doi.org/10.1016%2fj.mycres.2007.08.001 larsson k.-h., larsson e., köljalg u. 2004. high phylogenetic diversity among corticioid homobasidiomycetes. mycol res. 108: 983-1002. http://dx.doi.org/10.1017%2fs0953756204000851 larsson k.-h., parmasto e., fischer m., langer e., nakasone k.k., redhead s.a. 2006. hymenochaetales: a molecular phylogeny for the hymenochaetoid clade. mycologia 98: 926-936. http://dx.doi.org/10.3 852%2fmycologia.98.6.926 miettinen o., larsson k.-h. 2011. sidera, a new genus in hymenochaetales with poroid and hydnoid species. mycol. progress 10: 131-141. miettinen o., rajchenberg m. 2012. obba and sebipora, new polypore genera related to cinereomyces and gelatoporia (polyporales, basidiomycota). mycol. progress 11: 131-147. nakasone k.k., burdsall h.h. 2012. tsugacorticium kenaicum (hymenochaetales, basidiomycota), a new corticioid genus and species from alaska. north american fungi 7 (1): 1-9. ostrow h., dämmrich f. 2010. corticioid basidiomycetes in germany. z. mycol. 76(2): 177-210. ryvarden l., stokland j., larsson k.-h. 2003. a critical checklist of corticioid and poroid fungi of norway. synopsis fungorum 17: 1-79. sesli e., denchev c.m. 2008. checklists of the myxomycetes, larger ascomycetes, and larger basidiomycetes in turkey. mycotaxon 106: 65–67 + online version [2013-01]: 1-145 http://www.mycotaxon. com/resources/checklists/sesli-v106-checklist.pdf tellería m.t., dueñas m., beltrán-tejera e., rodríguez-armas j.l., martín m. 2012. a new species of hyphoderma (meruliaceae, polyporales) and its discrimination from closely related taxa. mycologia 104 (5): 1121-1132. tzean s.s., liou j.y. 1993. nematophagous resupinate basidiomycetous fungi. phytopathology 83: 1015-1020. wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków, 812 pp. lawrynomyces, a new genus of corticioid fungi 11 lawrynomyces – nowy rodzaj grzybów kortycjoidalnych w rzędzie hymenochaetales streszczenie w pracy opisano nowy rodzaj lawrynomyces karasiński gen. nov., dla hyphoderma capitatum. typ rodzaju jest gatunkiem notowanym głównie z europy, stwierdzonym także w azji mniejszej i nieznanym z innych kontynentów. występuje na mocno rozłożonym drewnie iglastym, głównie picea abies, rzadziej pinus sylvestris, w lasach mieszanych i iglastych, zarówno naturalnych jak i sztucznego pochodzenia, a owocniki wytwarza zazwyczaj późną jesienią. nowy rodzaj należy do tzw. „kladu rickenella” w rzędzie hymenochaetales. cechami charakterystycznymi dla lawrynomyces są rozpostarte, miękko błonkowate i białawe do szaro kremowych owocniki, bardzo cienkie (60–150 μm), tworzące na drewnie nieregularne w zarysie powłoczki (fig. 2 i 3), gładki hymenofor, monomityczny system strzępkowy ze strzępkami generatywnymi bez sprzążek, obecność cystyd tramalnych i hymenialnych oraz hyfid, podstawki niewyraźnie urnokształtne do niemal cylindrycznych z trzoneczkowatą podstawą oraz zarodniki szeroko elipsoidalne do prawie kulistych z grubiejącymi ścianami (fig. 1), nie wykazujące reakcji w odczynniku melzera i roztworze błękitu bawełnianego. w blisko spokrewnionym z lawrynomyces rodzaju peniophorella, podstawki mają kształt maczugowaty, zarodniki są cienkościenne, a należące tu gatunki często wytwarzają na strzępkach kontekstu charakterystyczne struktury – echinocysty i sferocysty, nieobecne u lawrynomyces. umieszczenie hyphoderma capitatum w odrębnym rodzaju ma uzasadnienie w publikowanych danych molekularnych. 2013-06-22t22:42:14+0100 polish botanical society new records of microbotryum species parasitizing caryophyllaceae from ukraine kyrylo g. savchenko1,2 and vasyl p. heluta2 1institute of evolution and faculty of natural sciences, university of haifa, mt. carmel, 31-905 haifa, israel, savchenko.kyryll@gmail.com 2m.g. kholodny institute of botany of the national academy of sciences of ukraine 2tereschenkivska st., ua-01601 kyiv, vheluta@botany.kiev.ua savchenko k. g., heluta v. p: new records of microbotryum species parasitizing caryophyllaceae from ukraine. acta mycol. 45 (2): 163–167, 2010. four records of smut fungi belonging to the genus microbotryum lév. are reported. two species were found on new hosts, namely m. dianthorum on dianthus borbasii and d. pseudoserotinus and m. superbum on d. stenocalyx. microbotryum lagerheimii on lychnis viscaria is a new species for ukraine. key words: smut fungi, microbotryum, dianthus, lychnis introduction the comprehensive information regarding european smut fungi has been given in the monograph by vánky (1994), where the author listed 400 species known for the continent and 70 provisory species. however, some new species have been described later and species known in other regions have been found here. thus, in the new check-list of smut fungi (vánky 2005a) 14 additional species were included. currently, there are 38 species which were described or collected after 1994 in europe (almaraz, telleria 1998; bauer et al. 2008; denchev 2007; denchev, giraud and hood 2009; lutz et al. 2005, 2008; lutz, vánky 2009; piątek 2006; prillinger et al. 2009; vánky 1998, 2003, 2005a,b, 2007, 2008a,b, 2009; vánky and berner 2003; vánky, horita and jage 2005a; vánky, jage and scholz 2005b; vánky et al. 2008; vánky, lutz 2007; vánky, scholz 2001). among them there are also representatives of the genus microbotryum lév., mainly described as new species, e.g., m. adenopetalae m. lutz, kemler et chleb., m. cartusianorum denchev, giraud et m.e. hood, m. chloranthae-verrucosum m. lutz, göker, piątek, kemler, begerow et oberw., m. lagerheimii denchev, m. minuartiae m. lutz, piątek, kemler et chleb., m. saponariae m. lutz, göker, piątek, kemler, acta mycologica vol. 45 (2): 163–167 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 164 k. g. savchenko and v. p. heluta begerow et oberw., m. schykoffianum giraud, denchev et m.e. hood, m. silenesacaulis m. lutz, piątek, kemler et chleb., m. silenes-dioicae giraud, denchev et m.e. hood, and m. silybum vánky et berner (denchev 2007; denchev et al. 2009; lutz et al. 2005, 2008; vánky, berner 2003). microbotryum jehudanum (zundel) vánky was found for the first time in the territory of europe (vánky et al. 2005b). another species well known as ustilago superba liro was transferred into the genus microbotryum with creation of a new combi nation m. superbum (liro) denchev, giraud et m.e. hood (denchev et al. 2009). it is known that many species of the genus microbotryum develop in the anthers of plants belonging to the caryophyllaceae. they are the so-called “anthericolous smut fungi”. these species easily produce hybrid forms which can subsequently become new species (chlebicki, suková 2005). therefore, for each region the establishment of the host-plants ranges of these species thorough morphological and, when it is possible, molecular investigations are all important. ukraine is a region with a considerable diversity of vascular plants, including the caryophyllaceae with the level of endemism at about 24% (fedoronchuk 2009). so, during the studies on smut fungi both rare species belonging to the genus microbotryum and new species of their host-plants may be found. this paper deals with such fungi which we have found on the lychnis l. and dianthus l. species in 2009. materials and methods sori and spores were studied using dried herbarium specimens. for light microscopy (lm), spores were dispersed in a droplet of lactophenol on a microscope slide, covered with a cover glass, gently heated to boiling point to rehydrate the spores and eliminate air bubbles, and examined at 400× and 1000× magnification. for scanning electron microscopy (sem), spores were placed on double-sided adhesive tape, mounted on a specimen stub, sputter-coated with gold, ca. 20 nm, and examined in sem at 30 kv. the studied materials are stored in the national herbarium of ukraine of m.g. kholodny institute of botany (kw). results and discussion smut fungi on dianthus species. for a long time, the smut fungi which develop their sori in the anthers of different dianthus species were included in ustilago violacea pers.: pers., a combined cosmopolitan parasite of hosts belonging to the caryophyllaceae. however, in 1924 liro described two separate species, u. dianthorum liro in the anthers of d. deltoides l. and u. superba liro on d. superbus l. much later, a new genus microbotryum was offered for some anthericolous smut fungi, and u. violacea s. lat. was included in this genus as m. violaceum (pers.) g. deml et oberw. (deml, oberwinkler 1982). scholz and scholz (1988) transferred smut fungi that fig. 1. spores of microbotryum dianthorum: a, b – on dianthus borbasii; c, d – on dianthus pseudoserotinus. all photos in sem. scale bars for a, d = 2 μm; b = 1 μm; c = 5 μm. fig. 2. microbotryum superbum on dianthus stenocalyx: a – infected flower, b, c – spores. microbotryum lagerhemii denchev on lychnis viscaria: d – infected flowers, e, f – spores; b, c, e, f – in sem. scale bars for a, d = 5 mm; b = 5 μm; c = 2 μm; e, f = 1 μm. new records of microbotryum 165 developed in anthers of many dianthus species into m. dianthorum (liro) h. scholz et i. scholz, but liro’s u. superba was left in m. violaceum. denchev and scharkova (1997), having investigated 229 specimens of smut fungi on 22 dianthus species, concluded that all of them belong to the same species, namely m. violaceum s. lat. however, vánky (2004) supposed that this species can be divided into smaller ones through the use of analyses of as many of type specimens as possible with employment of both traditional morphological and molecular-phylogenetic methods. such studies have been made by lutz et al. (2005, 2008), and their results confirmed the separation of m. dianthorum. however, within this species a great variability in the spore dimensions was found. in addition, the number of meshes per spore diameter also strongly varied. in spite of this, all specimens were clustered together by molecular data. similar investigations regarding other fungi from m. violcaeum-complex parasitizing d. sylvestris wulfen and d. cartusianorum l. showed insignificant morphological differences between both of these samples and from other specimens of this complex. however, they differed by molecular-biological data and features of microsatellite analyses. thus, they were cryptic species (denchev et al. 2009). the results of molecular-phylogenetic investigations also made it possible to restore a species status of smut fungus on d. superbus and regard it not any more as u. superba but as m. superbum (denchev et al. 2009). only two species of smut fungi parasitizing dianthus species were known in ukraine, e.g., m. dianthorum (reported as u. dianthorum) on d. pseudoarmeria m. bieb. from the forest-steppe zone and m. superbum (reported as u. superba) on d. superbus from polissia region (zerova et al. 1971). no specimens of these fungi were found in the national herbarium of ukraine of the m.g. kholodny institute of botany (fungi of ukraine 1996). so far, the smut fungi on dianthus have been considered to be rare in ukraine. however, in summer 2009 we collected three specimens of the anthericolous smut fungi on d. borbasii vandas, d. pseudoserotinus błocki (d. arenarius p.p.) and d. stenocalyx juz. (d. superbus p.p.). the first and the second of them were identified by us as m. dianthorum and the last as m. superbum. the characteristics of these fungi on new hosts are given below. microbotryum dianthorum (liro) h. scholz et i. scholz fig. 1 sori develop in the anthers of host plants. spore mass powdery, from medium violet to brownish violet. spores globose or subglobose, sometimes ovoid to slightly elongated, 5-8 × 5-7 μm. spore wall reticulate, on dianthus borbasii 5-7 meshes per spore diameter, meshes irregularly polygonal to rounded, 0.6-0.9 × 0.8-1 μm; on d. pseudoserotinus (6) 7-9 meshes per spore diameter, meshes rounded to slightly irregularly polygonal, 0.6 × 0.7 μm. distribution in ukraine. on dianthus borbasii vandas: cherkasy region, kaniv distr., trakhtemyriv regional landscape park, 49˚59’10’’ n, 31˚22’20’’ e, 03.06.2009, leg. k. savchenko (kw 36849f). on d. pseudoarmeria m. bieb.: rightbank forest-steppe zone (zerova et al. 1971). on d. pseudoserotinus błocki: volhynian region, lubeshiv distr., pripyat-stokhid national park, 51˚53’16’’ n, 24˚57’26’’ e, 09.08.2009, leg. k. savchenko and v. heluta (kw 36851f). microbotryum superbum (liro) denchev, giraud et m.e. hood figs 2 a-c sori develop in the anthers of host plants. spore mass dark violet brown, powdery. spores subglobose, ovoid to slightly elongated, sometimes rather irregular, 166 k. g. savchenko and v. p. heluta 5-6 (6.5) × 4-6 μm (at the mean 5.6 × 4.7 μm). spore wall reticulate, 5–7 meshes per spore diameter, meshes slightly irregular polygonal, 0.7 × 0.8 μm. distribution in ukraine. on dianthus stenocalyx juz.: volhynian region, lubeshiv distr., pripyat-stokhid national park, mixed forest, 51˚53’40’’ n, 24˚57’35’’ e, 08.08.2009, leg. v. heluta (kw 36852f). on d. superbus l. s. lat.: right-bank and left-bank polissia (zerova et al. 1971). microbotryum superbum differs from m. dianthorum in having smaller spores and by the color of spore mass. anthericolous smut fungus on lychnis. during field research in cherkasy region (ukraine) intense development of smut fungus in the anthers of lychnis viscaria l. was recorded. thus, about 20% of plants in the studied population were affected. the fungus was identified as microbotryum lagerheimii. this species has been recently described (denchev 2007) in the anthers of l. alpina l. and l. viscaria. it is closely related to m. silenes-inflatae (dc. ex liro) g. deml et oberw. known on lychnis alpina, silene vulgaris subsp. glareosa (jord.) marsden-jones et turrill, s. vulgaris subsp. vulgaris (moench) garcke (oberna behen (l.) ikonn.), s. cucubalus wibel, s. inflata sm. and s. venosa asch.) but differs in having far less (small or medium) intensity of spore mass coloration (denchev, minter 2008). microbotryum lagerhemii is also known from czech, finland, germany, italy, latvia, norway, poland, russia, and sweden (denchev 2007). thus, the fungus is a european endemic within a boreal-temperate area. in ukraine it was found for the first time. microbotryum lagerheimii denchev figs 2 d-f sori develop in the anthers of host plant. spore mass pale to medium violet, powdery. spores globose, subglobose or slightly ellipsoidal to irregular, 4.5-8 × 4.5-7 μm, length/width ratio 1.06–1.18, practically hyaline to weakly coloured. spore wall reticulate, 5-8 meshes per spore diameter, meshes globose to ellipsoidal and irregularly elongated, 0.3-1.1 μm. distribution in ukraine. cherkasy region, kaniv distr., trakhtemyriv regional landscape park, 49˚59’06’’ n, 31˚20’53’’ e, 04.06.2009, leg. k. savchenko (kw 36366f). acknowledgments. the authors would like to express their deep thanks to dr. mykola fedoronchuk (kyiv, ukraine) for checking of dianthus species, to mr. jason somers for checking our english, and to mr. viktor novychenko for assistance with the sem photographs. references almaraz t., telleria t.m. 1998. on ustilago sparti (ustilaginales, basidiomycotina). mycotaxon 67: 495– 504. bauer r., lutz m., begerow d., piątek m., vánky k., bacigálova k., oberwinkler f. 2008. anther smut fungi on monocots. mycol. res. 112: 1297–1306. chlebicki a., suková m. 2005. two microbotryum species from himalayas. mycotaxon 93: 149–154. deml g., oberwinkler f. 1982. studies in heterobasidiomycetes. part 24. on ustilago violacea (pers.) rouss. from saponaria officinalis l. phytopath. zeitschr. 104: 345–365. denchev c.m. 2007. microbotryum lagerheimii sp. nov. 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(ustilaginomycetes) from germany. mycol. balcan. 5: 79–81. vánky k., scholz h. 2001. three new species of smut fungi (ustilaginomycetes). nova hedwigia 72: 391–398. zerova m.ya, morochkovskyi s.f., radziyevskyi g.g., smitska m.f. 1971.vyznachnyk grybiv ukrainy. tom iv. basydiomitsety: dakrymitsetalni, tremelalni, auricularialni, sazhkovydni, irzhasti. kyiv, naukova dumka. 2014-01-01t11:51:11+0100 polish botanical society 2014-09-06t20:23:58+0200 polish botanical society lichens and allied fungi of two regional parks in vilnius area (lithuania) jurga motiejūnaitė laboratory of mycology, institute of botany, žaliųjų ežerų str. 49 lt-08406 vilnius 21, mikojm@botanika.lt motiejūnaitė j.: lichens and allied fungi of two regional parks in vilnius area (lithuania). acta mycol. 44 (2): 185–199, 2009. two regional parks – verkiai and pavilniai are situated in vilnius city area comprising territories both of the city itself and vilnius district and are under strong anthropogenic influence. during the present study 172 species of lichens, lichenicolous and saprobic fungi were revealed in verkiai rp and 92 species – in pavilniai rp. three lichen species – absconditella pauxilla, thelenella pertusariella (in verkiai rp), bacidia caligans (in pavilniai rp) and one saprobic fungus – chaenothecopsis debilis (in verkiai rp) are reporded for the first time in lithuania. seven lichen species of lithuanian red data book were recorded, all of them only in verkiai rp; this indicates less strong anthropogenic impact and better conditions for biodiversity in this park. key words: lichens, lichenicolous fungi, protected areas, biodiversity, lithuania introduction lichens of vilnius environs, though being closest to the centres of botanical investigations of lithuania – vilnius university and institute of botany was quite understudied up to present. first data on lichens from vilnius and environs, albeit very fragmentary, are found already in the first lithuanian botanical references: b.s. jundziłł (1811) and j. jundziłł (1822a, b, c), though in truth not a single territory in vilnius city or its environs were ever lichenologically detailly studied. this refers to the present areas of two regional parks situated in vilnius city territory: verkiai and pavilniai rp. none whatsever lichenological material was tracked for pavilniai rp, except single specimen of peltigera rufescens (weiss) humb. from wi herbarium cited by motiejūnaitė and miądlikowska (1998). first data on lichens from verkiai rp are found in mowszowicz (1957–1959). in total he indicates 43 lichen species, though it is not clear whether they are from the exact territory of the rp, the locality acta mycologica vol. 44 (2): 185–199 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 186 j. motiejūnaitė is indicated as wilno, werki. part of this report is dubious, e. g. opegrapha atra pers., bacidia obscurata (sommerf.) zahlbr. (mycobilimbia tetramera), cladonia glauca flörke, ramalina pollinaria (westr.) ach., but none of the herbarium specimens has survived to present time and it is impossible to revise them. during the last 15 years fragmentary data on lichen finds in verkiai rp (mainly žalieji ežerai landscape preserve) were published by several authors: balevičius (1992), kukwa & motiejūnaitė (2005), miądlikowska & motiejūnaitė (1994), motiejūnaitė (1999, 2007a), motiejūnaitė & miądlikowska (1998) and motiejūnaitė & prigodina (1999). this paper presents first detailed study of lichen biota in both verkiai and pavilniai regional parks. study area pavilniai regional park is situated at the southeastern edge of vilnius city (fig. 1). it occupies 2153 ha area, which includes territories of both vilnius city and vilnius district (54o39’–43’n, 25o19’–25’e). this smallest regional park in lithuania was established in 1992 (since 1984 this was territory of the nature park) to protect fig. 1. location of verkiai (1) and pavilniai (2) regional parks (map from state cadastre of protected areas: http://stk.vstt.lt, modified). lichens in vilnius 187 landscape of eroded ravine, hill system and valley of vilnia river. though part of the park is vilnius city, there are still numerous close-to-natural or seminatural forests with large diversity of vascular plants (forest covers 76,5 % of the park area). 10 preserves and one strict nature reserve warrant protection of rare plant species and communities as well as landscape formations. especially valuable is the area of kalnai strict nature reserve with diverse forest stand communities occupying relief with hills and ravines. verkiai regional park is situated at the northeastern edge of vilnius city (fig. 1), it occupies 2673 ha area, which includes territories of both vilnius city and vilnius district (54o44’–50’n, 25o17’–22’e). the park was established in 1992 to protect landscape of žalieji ežerai lakes’ complex and valuable cultural complexes of verkiai, kalvarijos and trinapolis. forest covers 76 % of the park area. southern part of the park comprises mainly cultural objects; meanwhile northern is rich in nature values. especially valuable is forest-covered hilly area with five glacial lakes, which are called žalieji (“green”) due to greenish colour of water with high content of carbonates. calcareous soils, beneficial microclimate and hilly relief ensure existence of exceptionally rich plant diversity (kirstukas 2004). both regional parks are under strong anthropogenic influence: some part of both comprises built-up areas, both parks are popular recreation zones for city people. besides, pavilniai park is strongly influenced by the emissions of vilnius city transport and industrial objects. material and methods lichens and allied fungi were collected in 2006 from the whole territory of verkiai and pavilniai regional parks. all habitats and substrates were considered. lichens were identified following routine microscopic techniques; sterile corticolous lichens as well as some specimens of cladonia genus were identified employing tlc (methodology following orange et al. 2001). critical specimens were compared with the identified collections at the herbaria of the botanical museum, university of copenhagen (c) and of the botanical museum berlin-dahlem (b). voucher specimens of the study are deposited in the herbarium of the institute of botany (vilnius) (bilas). list of species new species to lithuania are marked with !, lichenicolous fungi are marked with #, non-lichenized saprobic fungi are marked with +. nomenclature basically follows santesson et al. (2004), also further sources such as blanco et al. (2004), lücking et al. (2004), veldkamp (2004) and others. 188 j. motiejūnaitė verkiai regional park !absconditella pauxilla vězda & vivant – on dead mosses growing at the edge of pine stand; a. lignicola vězda & pišut – on wood of fallen, decaying, barkless trunk of picea abies. acarospora fuscata (schrad.) th. fr.– on a siliceous stone. #acremonium sp. – on primary squamules of cladonia coniocraea, possibly associated with nectriopsis parmeliae (for the description and notes see motiejūnaitė (2006)). acrocordia gemmata (ach.) a. massal. – on trunks of acer platanoides, quercus robur and populus tremula. amandinea punctata (hoffm.) coppins & scheid. – on trunks of quercus robur and acer platanoides. anaptychia ciliaris (l.) körb. – on trunks of acer platanoides, tilia cordata and quercus robur, on branches of populus tremula. anisomeridium polypori (ellis & everh.) m. e. barr – on trunk of acer platanoides. arthonia byssacea (weigel) almq. – on trunks of quercus robur and fraxinus excelsior; a. radiata (pers.) ach. – on trunks of fraxinus excelsior, on branches of corylus avellana; a. ruana a. massal. – on trunks of alnus glutinosa, quercus robur, fraxinus excelsior and acer platanoides, on branches of corylus avellana; a. spadicea leight. – on trunks of alnus glutinosa, a. incana, quercus robur and tilia cordata. #athelia arachnoidea (berk.) jülich – on thalli of various epiphytic lichens. bacidia rubella (hoffm.) a. massal. – on trunks of acer platanoides; b. subincompta (nyl.) arnold – on trunks of acer platanoides and alnus glutinosa. bacidina arnoldiana (körb.) v. wirth & vězda – on trunks of alnus glutinosa and alnus incana; b. inundata (fr.) vězda – on seasonally submerged roots of ulmus sp. by the stream side. biatora chrysantha (zahlbr.) printzen – on trunk of tilia cordata; b. ocelliformis (nyl.) arnold – on trunk of fraxinus excelsior. biatoridium monasteriense j. lahm ex körb. – on epiphytic mosses and bark on trunk base of acer platanoides. buellia griseovirens (turner & borrer ex sm.) almb. – on trunks and branches of various deciduous trees. calicium glaucellum ach. – on wood of living trunk of quercus robur. caloplaca citrina (hoffm.) th. fr. – on stone wall. candelariella reflexa (nyl.) lettau – on trunk of alnus glutinosa; c. vitellina (hoffm.) müll. arg. – on trunks of padus avium, quercus robur and populus tremula, on siliceous stones; c. xanthostigma (ach.) lettau – on trunks and branches of quercus robur, acer platanoides and populus tremula. catillaria nigroclavata (nyl.) schuler – on trunks of acer platanoides and populus tremula. cetrelia olivetorum (nyl.) w. l. culb. & c. f. culb. – on trunk of tilia cordata, on branches of corylus avellana and acer platanoides. note. the species is given here as cetrelia olivetorum s.l., as it was done so far in lithuania. studies aiming to reveal composition of the complex are pending. lichens in vilnius 189 chaenotheca brachypoda (ach.) tibell – on snags of picea abies; ch. chrysocephala (turner ex ach.) th. fr. – on trunks of picea abies; ch. ferruginea (turner ex sm.) mig. – on trunks of betula pendula, pinus sylvestris and tilia cordata; ch. furfuracea (l.) tibell – on roots of upended picea abies, on wood of quercus robur in a trunk hollow, on snags and stumps of picea abies; ch. phaeocephala (turner) th. fr. – on trunk of quercus robur; ch. stemonea (ach.) müll. arg. – on wood of snags, on trunks of picea abies and quercus robur; ch. trichialis (ach.) th. fr. – on trunks of quercus robur, on snags of picea abies; ch. xyloxena nádv. – on wood of snags. !+chaenothecopsis debilis (sm.) tibell – on wood of living trunk of quercus robur; +ch. pusilla (ach.) a. f. w. schmidt – on roots of upended picea abies. chrysothrix candelaris (l.) j. r. laundon – on trunk of quercus robur. cladonia cenotea (ach.) schaer. – on decaying stumps; c. chlorophaea (flörke ex sommerf.) spreng. – on trunk bases of pinus sylvestris, quercus robur and acer platanoides, on decaying wood; c. coniocraea (flörke) spreng. – on decaying wood, on trunk bases of various trees; c. digitata (l.) hoffm. – on decaying stumps, on trunk bases of pinus sylvestris; c. fimbriata (l.) fr. – on decaying stumps, on trunks of betula spp. on soil at the forest edge; c. grayi g. merr. ex sandst.. – on wood of fallen decaying trunks; c. homosekikaica nuno – on trunk of tilia cordata; c. macilenta hoffm. – on trunks of betula pendula and pinus sylvestris; c. ochrochlora flörke – on decaying wood of fallen trunks, snags and stumps; c. parasitica (hoffm.) hoffm. – on stump of quercus robur. #clypeococcum hypocenomycis d. hawksw. – on thalli of hypocenomyce scalaris. coenogonium pineti (ach.) lücking & lumbsch – on trunks of alnus glutinosa and pinus sylvestris. evernia prunastri (l.) ach. – on trunks and branches of various trees. fellhanera gyrophorica sérus., coppins, diederich & scheid. – on trunk of quercus robur. fuscidea pusilla tønsberg – on trunks of alnus glutinosa and betula pendula. graphis scripta (l.) ach. – on trunks of various deciduous trees. hypocenomyce scalaris (ach.) m. choisy – on decaying stumps and snags, on trunks of betula pendula and pinus sylvestris. hypogymnia physodes (l.) nyl. – on trunks and branches of various trees; h. tubulosa (schaer.) hav. – on trunks of tilia cordata. hypotrachyna revoluta (flörke) hale – on trunk of alnus glutinosa. #illosporium carneum fr. – on thallus of peltigera didactyla. imshaugia aleurites (ach.) s. l. f. meyer – on trunks of pinus sylvestris. lecania cyrtella (ach.) th. fr. – on trunk of salix sp.; l. hyalina (fr.) r. sant. – on trunks of tilia cordata and alnus incana; l. naegelii (hepp) diederich & p. boom – on branches of tilia cordata. lecanora allophana nyl. – on trunks and branches of populus tremula, on trunk of acer platanoides; l. argentata (ach.) malme – on trunk of acer platanoides; l. carpinea (l.) vain. – on trunks of tilia spp., acer platanoides and alnus glutinosa, on branches of populus tremula; l. chlarotera nyl. – on trunks of acer platanoides, tilia cordata and quercus robur, on branches of populus tremula; l. conizaeoides nyl. ex cromb. – on trunks of pinus sylvestris, on branches of betula sp.; l. expallens ach. – on trunks of quercus robur and acer platanoides; l. hagenii (ach.) 190 j. motiejūnaitė ach. – on trunk of quercus robur; l. pulicaris (pers.) ach. – on trunks of padus avium, quercus robur and fraxinus excelsior, on branches of picea abies; l. saligna (schrad.) zahlbr. – on snag of pinus sylvestris; l. symmicta (ach.) ach. – on branches of quercus robur and betula pendula; l. thysanophora r. c. harris – on trunks of tilia cordata and quercus robur; l. varia (hoffm.) ach.– on trunk of alnus glutinosa, on timber fence. lecidea nylanderi (anzi) th.fr. – on trunks of picea abies. lecidella elaeochroma (ach.) choisy – on trunks and branches of various deciduous trees; l. euphorea (flörke) hertel – on trunks of salix sp., acer platanoides and populus tremula, on branches of corylus avellana; l. flavosorediata (vězda) hertel & leuckert – on trunk of tilia cordata; l. stigmatea (ach.) hertel & leuckert – on stone wall; l. subviridis tønsberg – on trunk of quercus robur. lepraria incana (l.) ach. – on trunks of picea abies, alnus glutinosa, pinus sylvestris and betula spp., on decaying wood; l. jackii tønsberg – on trunk of pinus sylvestris; l. lobificans nyl. – on trunks of various deciduous trees. +leptorhaphis epidermidis (ach.) th. fr. – on trunk of betula pendula. #lichenoconium lecanorae (jaap) d. hawksw. – on apothecia of lecanora carpinea; #l. erodens m. s. christ. & d. hawksw. – on thalli of melanohalea exasperatula, ramalina farinacea and parmelia sulcata; #l. usneae (anzi) d. hawksw. – on thallus of melanohalea exasperatula. lobaria pulmonaria (l.) hoffm. – on trunks of acer platanoides and tilia cordata. #marchandiomyces aurantiacus (lasch) diederich & etayo – on thalli of physcia tenella, physcia stellaris, xanthoria parietina and x. polycarpa. melanelixia fuliginosa (fr. ex duby) o. blanco et al. – on trunks of alnus glutinosa, acer platanoides and tilia cordata; m. subargentifera (nyl.) o. blanco et al. – on trunk of tilia cordata; m. subaurifera (nyl.) o. blanco et al. – on branches of picea abies, on trunk of acer platanoides. melanohalea exasperatula (nyl.) o. blanco et al. – on trunks and branches of various trees. micarea denigrata (fr.) hedl. – on timber fence; m. hedlundii coppins – on decaying stump of quercus robur; m. melaena (nyl.) hedl. – on trunks of pinus sylvestris; m. micrococca (körb.) gams ex coppins – on decaying stumps, on trunks of populus tremula and pinus sylvestris; m. misella (nyl.) hedl. – on decaying stumps and wood of fallen trees; m. prasina fr. – on wood of fallen trees, on trunks of tilia cordata, pinus sylvestris, alnus glutinosa and acer platanoides. +microcalicium ahlneri tibell – on soft decaying wood in a hollow of quercus robur trunk. #m. disseminatum (ach.) vain. – on trunks of quercus robur, apparently on dead thalli of chaenotheca spp. mycoblastus fucatus (stirt.) zahlbr. – on trunks of betula pendula, on twigs of picea abies. +mycocalicium subtile (pers.) szatala – on snag of pinus sylvestris. #nectriopsis parmeliae (berk & m. a. curtis) m. s. cole & d. hawksw. – on primary squamules of cladonia sp. nephroma parile (ach.) ach. – on epiphytic mosses on trunk of old acer platanoides. ochrolechia androgyna (hoffm.) arnold s.l.– on trunks of populus tremula and quercus robur. lichens in vilnius 191 note. detailed discussion on ochrolechia genus in lithuania see in kukwa (2009). ochrolechia microstictoides räsänen – on trunks of quercus robur. opegrapha rufescens pers. – on trunks of acer platanoides and fraxinus excelsior; o. varia pers. – on trunks of acer platanoides, quercus robur and ulmus sp. parmelia submontana nádv. ex hale – on branches of corylus avellana; p. sulcata taylor – on trunks and branches of various trees. parmelina tiliacea (hoffm.) hale – on branches of quercus robur and populus tremula, on trunks of tilia spp. parmeliopsis ambigua (wulfen) nyl. – on trunks of betula pendula, pinus sylvestris and alnus glutinosa. peltigera didactyla (with.) j. r. laundon – on soil at forest edges and road scarps, on mosses growing on stone wall; p. praetextata (flörke ex sommerf.) zopf – on soil at the forest edge, on bases of deciduous trees, on moss-covered fallen trees and stumps; p. rufescens (weiss) humb. – on soil at the edge of pinus sylvestris stand. pertusaria albescens (huds.) m. choisy et werner – on trunks of various deciduous trees; p. amara (ach.) nyl. – on trunks of various deciduous trees; p. coccodes (ach.) nyl. – on trunks of tilia cordata, acer platanoides and populus tremula; p. leioplaca dc. – on trunks of fraxinus excelsior and tilia cordata, on branches of corylus avellana. phaeophyscia endophoenicea (harm.) moberg – on trunk of acer platanoides; ph. orbicularis (neck.) moberg – on trunks and branches of various deciduous trees, on concrete. phlyctis agelaea (ach.) flot. – on trunks of acer platanoides; ph. argena (spreng.) flot. – on trunks of various trees. #phoma cytospora (vouaux) d. hawksw. – on thallus of hypogymnia physodes. physcia adscendens h. olivier – on trunks and branches of various deciduous trees; ph. stellaris (l.) nyl. – on branches of various deciduous trees; ph. tenella (scop.) dc. – on trunks and branches of various deciduous trees. physconia distorta (with.) j. r. laundon – on trunks of acer platanoides, on branches of populus tremula; ph. enteroxantha (nyl.) poelt – on trunks of tilia spp. and alnus glutinosa, on mosses growing on stone wall; ph. perisidiosa (erichsen) moberg – on trunks of quercus robur, acer platanoides and salix spp. placynthiella icmalea (ach.) coppins & p. james – on decaying stumps and wood of fallen trees. platismatia glauca (l.) w. l. culb. & c. f. culb. – on branches of various trees. pleurosticta acetabulum (neck.) elix & lumbsch – on trunks of acer platanoides. protoparmeliopsis muralis (schreb.) m. choisy – on siliceous stones. pseudevernia furfuracea (l.) zopf – on trunks and branches of various trees. pseudosagedia aenea (wallr.) hafellner & kalb – on trunks of populus tremula and fraxinus excelsior. pyrenula nitidella (flörke ex schaer.) müll. arg. – on trunk of acer platanoides. ramalina baltica lettau – on trunks of acer platanoides and quercus robur; r. farinacea (l.) ach. – on trunks of various trees; r. fastigiata (pers.) ach. – on trunks of acer platanoides and quercus robur, on branches of populus tremula; r. fraxinea (l.) ach. – on branches of populus tremula. 192 j. motiejūnaitė reichlingia leopoldii diederich & scheid. – on trunks of tilia cordata and quercus robur. rinodina pyrina (ach.) arnold – on branches of tilia cordata. rinodina exigua (ach.) gray – on trunk of acer platanoides. ropalospora viridis (tønsberg) tønsberg – on trunks of acer platanoides, quercus robur, padus avium, populus tremula, alnus glutinosa and betula spp., on branches of corylus avellana. #roseliniella cladoniae (anzi) matzer & hafellner – on thallus of cladonia chlorophaea. scoliciosporum umbrinum (ach.) arnold – on a siliceous stone. strangospora moriformis (ach.) stein – on timber fence. #syzygospora physciacearum diederich – on thallus of physcia stellaris. #taeniolella beschiana diederich – on thallus of cladonia chlorophaea. !thelenella pertusariella (nyl.) vain. – on base of old lonicera xylosteum branches, on branches of corylus avellana. trapelia placodioides coppins et p. james – on a siliceous stone. trapeliopsis flexuosa (fr.) coppins & p. james – on various decaying wood; t. granulosa (hoffm.) lumbsch – on decaying stumps, on trunks of pinus sylvestris. #tremella cladoniae diederich et m.s.christ. – on thalli of cladonia coniocraea; #t. lichenicola diederich – on thalli of mycoblastus fucatus. tuckermanopsis chlorophylla (willd.) hale – on trunk of tilia cordata. usnea subfloridana stirt. – on branches of tilia cordata. verrucaria dolosa hepp – on calcareous pebbles; v. hydrela ach. – on siliceous stones in a stream bed, on seasonally submerged roots of ulmus sp. by the stream side; v. muralis ach. – on calcareous pebbles; v. nigrescens pers. – on stone wall; v. praetermissa (trevis.) anzi – on siliceous stones in a stream bed. vezdaea aestivalis (ohlert) tscherm.-woess & poelt – on epiphytic mosses on trunks of tilia cordata and acer platanoides. xanthoparmelia conspersa (ach.) hale – on siliceous stones; x. stenophylla (ach.) hale – on siliceous stones. xanthoria parietina (l.) th. fr. – on trunks and branches of various deciduous trees, on concrete; x. polycarpa (hoffm.) th. fr. ex rieber – on branches, rarely on trunks of various deciduous trees. #xanthoriicola physciae (kalchbr.) d. hawksw. – on thallus and apothecia of xanthoria parietina. pavilniai regional park absconditella lignicola vězda et pišut – on wood of decaying fallen picea abies trunk. amandinea punctata (hoffm.) coppins et scheid. – on trunks of tilia cordata and acer platanoides. anisomeridium polypori (ellis et everh.) m. e. barr – on trunks of acer platanoides and ulmus sp. lichens in vilnius 193 arthonia byssacea (weigel) almq. – on trunk of acer platanoides; a. ruana a. massal. – on trunks of ulmus sp. and alnus incana. #athelia arachnoidea (berk.) jülich – on thalli of various epiphytic lichens. bacidia brandii coppins & p. boom – on twigs of vaccinium myrtillus; !bacidia caligans (nyl.) a. l. sm. – on branches of old sambucus racemosa. bacidina arnoldiana (körb.) v. wirth & vězda – on decaying wood of fallen trees, on trunks of ulmus sp. and alnus incana. “biatora” sp. – on branches of old sambucus racemosa, on trunks of ulmus sp., alnus incana and acer platanoides. note. this sterile sorediate species lacking any secondary metabolites was tentatively ascribed to biatora genus (for further notes on “biatora” sp. see motiejūnaitė (2007b), motiejūnaitė and jucevičienė (2005)). buellia badia (fr.) a. massal. – on a siliceous stone; b. griseovirens (turner & borrer ex sm.) almb. – on branches of corylus avellana and quercus robur, on trunks of tilia cordata and populus tremula, on wood of decaying fallen trunk of picea abies. calicium viride pers. – on trunks of quercus robur. caloplaca saxicola (hoffm.) nordin – on stone wall. candelariella reflexa (nyl.) lettau – on trunks of populus tremula; c. vitellina (hoffm.) müll. arg. – on a siliceous stone; c. xanthostigma (ach.) lettau – on trunks of quercus robur and ulmus sp. chaenotheca brachypoda (ach.) tibell – on wood of acer platanoides in trunk hollow; ch. chrysocephala (turner ex ach.) th. fr. – on trunk of tilia cordata; ch. ferruginea (turner ex sm.) mig. – on trunks of quercus robur, picea abies and pinus sylvestris; ch. phaeocephala (turner) th. fr. – on trunks of quercus robur; ch. stemonea (ach.) müll. arg. – on trunks of picea abies; ch. trichialis (ach.) th. fr. – on trunks of quercus robur and acer platanoides. cladonia coniocraea (flörke) spreng. – on bases of various deciduous trees, on decaying wood of stumps and fallen trees; c. fimbriata (l.) fr. – on bases of various deciduous trees, on decaying wood of stumps and fallen trees; c. furcata (huds.) schrader – on soil at the edge of pine stand; c. homosekikaica nuno – on stump of picea abies; c. macilenta hoffm. – on decaying stumps. #clypeococcum hypocenomycis d. hawksw. – on thalli of hypocenomyce scalaris. coenogonium pineti (ach.) lücking & lumbsch – on trunks of quercus robur and picea abies. evernia prunastri (l.) ach. – on trunks and branches of quercus robur and tilia cordata. fuscidea pusilla tønsberg – on trunks of populus tremula. graphis scripta (l.) ach. – on trunks of ulmus sp., alnus incana and quercus robur. hypocenomyce scalaris (ach.) m. choisy – on trunks of picea abies, pinus sylvestris, quercus robur and betula pendula. hypogymnia physodes (l.) nyl. – on trunks and branches of various trees; h. tubulosa (schaer.) hav. – on trunks of betula pendula. #illosporiopsis christiansenii (b. l. brady & d. hawksw.) d. hawksw. – on thalli of physcia tenella. 194 j. motiejūnaitė lecania cyrtella (ach.) th. fr. – on branches of corylus avellana, quercus robur and sambucus racemosa; l. naegelii (hepp) diederich & p. boom – on branches of sambucus racemosa. lecanora carpinea (l.) vain. – on trunks of quercus robur and acer platanoides; l. chlarotera nyl. – on trunks of quercus robur and acer platanoides; l. conizaeoides nyl. ex cromb. – on trunks of picea abies and betula pendula; l. expallens ach. – on trunks of quercus robur and tilia cordata; l. hagenii (ach.) ach. – on branches of fraxinus excelsior; l. polytropa (ehrh. ex hoffm.) rabenh. – on a siliceous stone; l. pulicaris (pers.) ach. – on trunks of picea abies and alnus incana, on branches of alnus incana and tilia cordata; l. symmicta (ach.) ach. – on trunks of quercus robur and acer platanoides. lecidella elaeochroma (ach.) choisy – on trunks and branches of various deciduous trees. lepraria incana (l.) ach. – on trunks of various trees; l. lobificans nyl. – on trunks of ulmus sp. +leptorhaphis epidermidis (ach.) th. fr. – on trunk of betula pendula. #lichenoconium erodens m. s. christ. & d. hawksw. – on thalli of hypogymnia physodes. #marchandiomyces aurantiacus (lasch) diederich & etayo – on thalli of physcia tenella, xanthoria parietina and x. polycarpa. melanohalea exasperatula (nyl.) o. blanco et al. – on branches of quercus robur, corylus avellana, malus domestica and picea abies. melanelixia fuliginosa (fr. ex duby) o. blanco et al. – on branches of corylus avellana, on trunks of tilia cordata, alnus incana, acer platanoides and quercus robur. micarea prasina fr. – on wood of decaying fallen trees. mycoblastus fucatus (stirt.) zahlbr. – on branches of corylus avellana. +mycocalicium subtile (pers.) szatala – on snag of picea abies. parmelia sulcata taylor – on trunks and branches of various deciduous trees, on siliceous stones. parmelina tiliacea (hoffm.) hale – on trunks of tilia cordata, quercus robur and acer platanoides, on branches of malus domestica and quercus robur. parmeliopsis ambigua (wulfen) nyl. – on trunks of betula pendula. peltigera praetextata (flörke ex sommerf.) zopf – on decaying fallen tree, laying in a stream bed. pertusaria albescens (huds.) m.choisy et werner – on trunks of quercus robur and tilia cordata. phaeophyscia nigricans (flörke) moberg – on trunks of populus tremula; ph. orbicularis (neck.) moberg – on trunks and branches of various deciduous trees, on siliceous stones. phlyctis argena (spreng.) flot. – on trunks of various deciduous trees. physcia adscendens h. olivier – on branches of various deciduous trees, on siliceous stones; ph. caesia (hoffm.) fürnr. – on a siliceous stone; ph.stellaris (l.) nyl. – on branches of corylus avellana, malus domestica, alnus incana and fraxinus excelsior; ph. tenella (scop.) dc. – on branches of various deciduous trees, on siliceous stones. physconia enteroxantha (nyl.) poelt – on trunks of tilia cordata and alnus incana. lichens in vilnius 195 placynthiella icmalea (ach.) coppins & p.james – on decaying wood of stumps and fallen trees. platismatia glauca (l.) w. l. culb. & c. f. culb. – on branches of picea abies. porpidia crustulata (ach.) hertel & knoph – on a siliceous stone. protoparmeliopsis muralis (schreb.) m. choisy – on a siliceous stone. pseudevernia furfuracea (l.) zopf – on branches of alnus incana, quercus robur and picea abies. pseudosagedia aenea (wallr.) hafellner & kalb – on trunk of alnus incana. +pycnidiella resinae (ehrenb.) höhn. – on resinous exudates of picea abies. ramalina farinacea (l.) ach. – on trunks of quercus robur; r. fastigiata (pers.) ach. – on trunks of alnus incana; r. fraxinea (l.) ach. – on trunks of quercus robur and ulmus sp. reichlingia leopoldii diederich & scheid. – on trunk of quercus robur. ropalospora viridis (tønsberg) tønsberg – on trunks of alnus incana and quercus robur. +stenocybe pullatula (ach.) stein – on trunks of alnus incana. trapeliopsis granulosa (hoffm.) lumbsch – on decaying wood of stumps and fallen trees. tuckermanopsis chlorophylla (willd.) hale – on branches of picea abies. verrucaria hydrela ach. – on siliceous stomes in a stream bed; v. praetermissa (trevis.) anzi – on siliceous stones in a stream bed. vulpicida pinastri (scop.) j.-e.mattsson & m. j. lai – on trunks of quercus robur and betula pendula. xanthoparmelia conspersa (ach.) hale – on a siliceous stone. xanthoria parietina (l.) th. fr. – on trunks and branches of various deciduous trees; x. polycarpa (hoffm.) th. fr. ex rieber – on branches of various deciduous trees. results and discussion altogether 172 species of lichens and allied fungi were recorded in verkiai rp and 92 species in pavilniai rp. three lichen species – absconditella pauxilla, thelenella pertusariella (from verkiai rp), bacidia caligans (from pavilniai rp) and one saprobic fungus – chaenothecopsis debilis (in verkiai rp) were recorded for the first time in lithuania. absconditella pauxilla is a rare species, so far mainly reported from western europe, also known from sweden (santesson et al. 2004), czech republic (palice 1999) and northern poland (czarnota, kukwa 2008). apparently it is mainly confined to dead bryophytes and more rarely to wood in humid situations. present find in lithuania confirms ecological requirements of this species earlier mentioned by other authors: the lichen inhabited patch of dead mosses growing on the lake shore. thelenella pertusariella is another rarely collected species, previously thought to be arctic-alpine. it is characterised by corticolous habit, pale, immersed perithecia, submuriform ascospores and asci with unthickened apex. in eurasia it is known so 196 j. motiejūnaitė far from austrian alps (mayrhofer 1987), fennoscandia – sweden, norway and finland (santesson et al. 2004), russia (novgorod and murmansk regions, northern urals and western siberia) (kataeva et al. 2005), latvia (piterāns 1982) and germany (cezanne, eichler 2002). the ecology of the lichen in temperate lowlands is little known, but apparently it favours shady humid habitats in old forests (e.g. description in cezanne, eichler 2002). t. pertusariella is doubtfully separated from a similar species thelenella vezdae (h. mayrhofer et poelt) coppins et fryday which differs in half-immersed to sessile perithecia, thin, richly branched and anastomosing paraphysoids, asci with thickened apex and predominantly lignicolous habit (mayrhofer, poelt 1985). recently, however, it is discussed that both taxa might be conspecific (cezanne, eichler 2002; palice 1999). bacidia caligans is being reported during the last 20 years with increasing frequency: it was found in a number of countries of northern, central and southern europe. very often it is recorded from anthropogenic substrates or habitats under strong anthropogenic influence, e. g. trees and shrubs in cities and towns (aptroot et al. 2005; diederich 1989; kowalewska, kukwa 2003), stone buildings and urban wasteland (coppins 1992; gilbert 1990; larsen 1995), therefore it is quite possible to assume that the lichen is spreading recently. in our case b. caligans was found on branches of sambucus racemosa in a suburban forest with low lichen diversity, apparently influenced by transport and industrial emissions of the city. chaenothecopsis debilis is a widely distributed saprobic calicioid fungus found on lignum of various trees (tibell 1999). of all species of the genus, which were recorded in lithuania, it is distinguished mainly by chemical reactions with k and hno3. c. debilis apparently is not common in lithuania, as it becomes more rare southwards of the boreal zone and, though not uncommon in fenoscandia, it was recorded only in few countries of central and southern europe (tibell 1999; titov 2006 and literature cited therein). during the present study 7 lichen species of lithuanian red data book (lrdb) were registered (all only in verkiai rp) (tab. 1): 2 species of category 1 (e): nephroma parile (1 locality), cladonia parasitica (1 locality); 3 species of category 2 (v): cetrelia olivetorum (2 localities), lobaria pulmonaria (2 localities), phaeophyscia endophoenicea (1 locality) and 2 species of category 3 (r): hypotrachyna revoluta (1 locality), ramalina baltica (2 localities). almost all their localities (except cladonia parasitica, one locality of cetrelia olivetorum and one locality of ramalina baltica) were situated in žalieji ežerai landscape preserve where fragments of old hardwood and hardwood-spruce forests are still present. a decade earlier three more lrdb species were found in verkiai rp, which where not registered during present study, maybe overlooked: arthonia didyma körb , chaenotheca chlorella (ach.) müll. arg. and lecanora albella (pers.) ach. (motiejūnaitė 1999, 2007a). table 1 data of diversity of lichens and allied fungi in verkiai and pavilniai regional parks regional park total number of species number of lrdb species number of indicators of biologically valuable species (excluding lrdb species) verkiai rp 172 7 16 pavilniai rp 92 – 4 lichens in vilnius 197 16 species of lichens, indicators of biologically rich forests (andersson et al. 2002; motiejūnaitė et al. 2004) were registered, meanwhile only four indicators were found in pavilniai rp (tab. 1). notably, the latter park not only bore lower numbers of rare and vulnerable species, but their abundance was visibly lower than in verkiai rp: e.g., arthonia byssacea was common in verkiai rp, meanwhile in pavilniai it was found only on one phorophyte; pseudosagedia aenea and chaenotheca brachypoda were also quite common in verkiai rp, but both were found only on one phorophyte every in pavilniai rp. chaenotheca phaeocephala, however, was uncommon in both parks. conclusions evaluation of diversity of lichen biota of verkiai and pavilniai regional parks shows strong differences due to different anthropogenic pressure induced on both territories. verkiai rp, at least in parts, has still preserved qualities of biocentre of lichens characteristic of old-growth forests, meanwhile pavilniai rp bear nothing more than common lichen biota of suburban areas. verkiai rp, especially žalieji ežerai landscape preserve is still a refugium for several very rare and vulnerable species in lithuania, such as nephroma parile and cladonia parasitica. vulnerable and rapidly declining lichen lobaria pulmonaria is still found in large and viable pupulations. the gravest dangers for the habitats of these lichens are transport and related air pollution as well as increasing stress of recreational activities. acknowledgements. it is a great pleasure and honour to dedicate this paper to my colleague and longtime friend and co-worker professor krystyna czyżewska with sincere thanks for her kindness and many warm moments shared together in poland and in lithuania. the inventory of lichen biota was financed by the directorate of verkiai and pavilniai regional parks. visit to the copenhagen university was granted by the eu cordis programme synthesys (dktaf-1825). i am grateful to ulrik søchting for providing possibility to use tlc techniques and to eric steen hansen for help while working at the herbarium (c). visit to the botanical museum berlin-dahlem (freie universität berlin) was granted by the eu cordis programme synthesys (de-taf-1905), curator of the herbarium harrie sipman is cordially thanked for the help while working there. 198 j. motiejūnaitė references aptroot a., czarnota p., jüriado i., kocourková j., kukwa m., lõhmus p., palice z., randlane t., saag l., sérusiaux e., sipman h., sparrius l. b., suija a., thüs h. 2005. new or interesting lichens and lichenicolous fungi found during the 5th ial symposium in estonia. folia crypt. estonica 41: 13–22. andersson l., kriukelis r., čiuplys r. 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(in:) nordic lichen flora 1: 20–94, bohuslän ‘5, uddevalla. titov a. n. 2006. mikokalicijevyje griby (poriadok mycocaliciales) golarktiki. kmk scientific press, moscow. veldkamp j. f. 2004. bilimbia (lichenes) resurrected. lichenologist 36: 191–195. 2014-01-01t11:49:25+0100 polish botanical society is small mammal mycophagy relevant for truffle cultivation? alexander urban1, marija kataržytė2, susanne schickmann3, katharina kräutler4 and tony pla1,5 1department of systematic and evolutionary botany, faculty of life sciences university of vienna, rennweg 14, a-1030 vienna, alexander.urban@univie.ac.at 2coastal research and planning institute, klaipėda university, lt-92294, klaipėda 3department of integrative biology and biodiversity research, institute of wildlife biology and game management, university of natural resources and life sciences, gregor-mendel-strasse 33, a-1180 vienna 4department of forest and soil sciences, institute of forest entomology, forest pathology and forest protection university of natural resources and life sciences, hasenauerstraße 38, a-1180 vienna 5 trüffelgarten urban & pla og, burwegstr. 88, a-3034 eichgraben urban a., kataržytė m., schickmann s., kräutler k., pla t.: is small mammal mycophagy relevant for truffle cultivation? acta mycol. 47 (2): 139–143, 2012. the role of mycophagous small mammals as vectors of hypogeous fungi is well established. however, little is known about dispersal of gourmet truffle species by mammal vectors, or about the potential role of mycophagy in truffle plantations. we hypothesize that small mammal mycophagy contributes to the productivity of truffle plantations by providing inoculum for truffle mycelium establishment and mating. spread of non-desired competitors of gourmet truffles is a potential adverse effect of small mammal mycophagy. keywords: tuber aestivum, tuber melanosporum, mutualism, symbiosis, mycorrhiza, ectomycorrhiza introduction hypogeous fungi and other macrofungi are part of the diet of small rodents (rodentia) such as voles (arvicolinae, cricetidae, myomorpha), mice (murinae, muridae, myomorpha), dormice (gliridae, sciuromorpha) and sqirrels (sciuridae, sciuromorpha) (maser, claridge and trappe 2008). recently it was found that insectivorous shrews (sorex spp., soricidae, eulipotyphla) frequently feed on hypogeous fungi as well (kataržytė, kutorga 2011; schickmann et al. 2012). the nutritional ecology of most small mammal species seems to be rather flexible, highly adaptive and more diverse than commonly assumed. spore dispersal in hypogeous fungi including acta mycologica vol. 47 (2): 139–143 2012 140 a. urban et al. gourmet truffle species depends entirely on animal vectors, and the mutualistic relationship between mycophagous animals and hypogeous fungi can be considered obligate for the latter. small mammal mycophagy was extensively studied in north america and australia, since it was recognized as a process potentially important for the maintenance of fungal and mammal biodiversity and for forest regeneration (claridge 2002; maser et al. 2008). in europe, relatively few studies demonstrated the role of mycophagy in bank vole, (myodes glareolus, blaschke and bäumler 1989), red squirrel (sciurus vulgaris, grönwall, pehrson 1984; bertolino et al. 2004) and wood mice (apodemus spp., blaschke, bäumler 1989). during a study conducted in primary and secondary forests in two regions of the eastern alps we found that all mammal species native to central europe mountainous forests investigated feed on hypogeous fungi, albeit with different intensity (schickmann et al. 2012). based on frequency, quantity and diversity of fungal spores detected in faeces by microscopy and with molecular tools, we concluded that at least one native small mammal species, the bank vole (myodes glareolus) is preferentially mycophagous. all other species captured, including wood mice (apodemus flavicollis and a. sylvaticus), other vole species, shrews (sorex spp.) and the fat doormouse (glis glis) were found to be opportunistically mycophagous. all small mammal species were found to feed on a diversity of fungal species. similar results were reported from hemiboreal forests in lithuania (kataržytė, kutorga 2011). methods here we synthesize available information to infer potential roles of small mammals in truffle plantations. a descriptinon of methods used for life trapping of small mammals, fecal pellet sample collection and microscopical analysis is found in (schickmann et al. 2012). a protocol optimized for dna extraction from fungal spores in fecal pellets of small mammals was developed (schickmann et al. 2011). results potential roles of small mammals in truffle plantations effects on host trees. small mammal communities differ with habitat types. rodent species are more or less herbivorous, and some species are known to damage host trees, at least at high population densities. red squirrels are likely to occur in mature plantations, especially if hazelnuts (corylus spp.) or conifer seeds are available. squirrels are very effective seed predators, hoarding of excess food in caches contributes to seed dispersal. sqirrels can damage trees by bark stripping, however, this behaviour is a major issue in the invasive grey squirrel (sciurus carolinensis) but not in the native red squirrel (bertolino, genovesi 2003). voles can cause severe damage in young truffle plantations. newly established truffle plantations in small mammal mycophagy 141 agricultural environmenments are more likely to be colonized by common voles (microtus arvalis) than by species ocurring in more natural habitats such as field vole (microtus agrestis) and bank vole (myodes glareolus). an invasion of voles into an experimental truffle plantation was observed when a neighbouring maize field was harvested and the animals lost shelter (ronald vogl, personal communication). debarking of the stem base killed one corylus colurna tree (4 yrs after outplanting). it appears that some tree species are less preferred by voles, e.g., pine species are avoided (borowski 2007). lime tree (tilia cordata) seedlings were reported to be preferentially attacked by bank voles (pigott 1985). planting a diversity of tree species may the best strategy to limit overall damage, given the diversity of potential pest species. according to our experience, voles are rarely a cause of excessive tree mortality in truffle plantations, but pressure can vary considerably. if the risk of loosing tree seedlings due to damage by voles is high, the roots and stem bases can be protected with a non-galvanized wire-basket which will decompose by corrosion. mulching can provide shelter for small mammals depending on the material used and should be avoided or adapted if pressure is critical. small mammal species are an important source of prey for predators such as various species of birds of prey, owls, red fox (vulpes vulpes), wildcat (felis silvestris), and various mustelids such as weasel (mustela nivalis), european polecat (mustela putorius) and european badger (meles meles). providing habitat for predators and protecting them from beeing hunted is an effective way to keep small mammal populations at levels compatible with plantation management objectives. according to our experience, game species, like european hare (lepus europaeus) and roe deer (capreolus capreolus) usually impose much higher pressure on young truffle plantations and need to be excluded. effects on truffle populations. little is known about effects of mycophagy on truffle populations in managed plantations. the small rodent species common in habitats like truffle plantations, e.g., voles, wood mice (apodemus syvaticus and a. flavicollis) and squirrels are well known to be mycophagous (maser et al. 2008). sqirrels even cache hypogeous fungi (vernes, poirier 2007). recently it was shown that also insectivores such as shrews (sorex spp.) frequently feed on hypogeous fungi (kataržytė, kutorga 2011; schickmann et al. 2012). this result is of interest for two reasons: 1) shrews are not herbivorous and they are very unlikely to harm host trees. 2) shrews are insectivorous and ground-dwelling, and may contribute to controlling insects parasitic on truffles or host tree roots. from the truffle growers perspective, mycophagy may be regarded as a waste of valuable crop. small mammals are important dispersal agents for truffle spores, but no data are available which proof that there is a role for mycophagist in plantations established with mycorrhized trees. once established, genets of truffle mycelium can grow and and extend, without apparent need of additional spores. however, three lines of evidence suggest that mycophagy may be essential for the long term fertility and productivity of truffle plantations: 1) truffle spores in faeces of small mammals are a viable source of inoculum (schickmann et al. 2012). 2) artificial inoculation of soil with truffle spores is reported to inrease yields of tuber aestivum (p. sourzat, unpublished). 3) sexual reproduction and outcrossing has been proven in some gourmet truffle species (paolocci et al. 2006; riccioni et al. 2008), and ascopore or 142 a. urban et al. conidiospore (urban et al. 2004) dispersal is likely to play a role in fertilisation of opposing mating types. truffle spore dispersal by small mammal vectors has one potential drawback: mycophagists use to feed on a variety of species of hypogeous fungi. thereby, they likely vector non-marketable or low-value species which may compete with the target gourmet truffle within the plantation. conclusions currently, information on the role of small mammal mycophagists in truffle plantations is scarce, despite the significance of mycophagy in the truffle life cycle. small mammals, at least if present in excess, are typically regarded as pests, by foresters and truffle-growers. some truffle growers use to combat small mammals by different means of pest control, but systematic experimentation is still lacking. at present we do not know whether the contribution of spore dispersal by small mammals to productivity is comparable to the relevance of pollination in fruit orchards. the impact of biotic interactions on productivity is more difficult to assess than the influence of abiotic factors, such as climate. experimental work on mycophagy in truffle plantation and natural truffle sites is needed to obtain reliable data on the role of small mammals in the dispersal and reproduction of truffle species. references bertolino s., genovesi p. 2003. spread and attempted eradication of the grey squirrel (sciurus carolinensis) in italy, and consequences for the red squirrel (sciurus vulgaris) in eurasia. biological conservation 109: 351–358. doi:10.1016/s0006-3207(02)00161-1. bertolino s., vizzini a., wauters l.a., tosi g. 2004. consumption of hypogeous and epigeous fungi by the red squirrel (sciurus vulgaris) in subalpine conifer forests. forest ecology and management 202: 227–233. doi:10.1016/j.foreco.2004.07.024. blaschke h. & bäumler. w. 1989. mycophagy and spore dispersal by small mammals in bavarian forests. forest ecology and management 26: 237–245. doi:10.1016/0378-1127(89)90084-4. borowski z. 2007. damage caused by rodents in polish forests. international journal of pest management 53: 303–310. doi:10.1080/09670870701497253. claridge a. w. 2002. ecological role of hypogeous ectomycorrhizal fungi in australian forests and woodlands. plant and soil 244: 291–305. doi:10.1023/a:1020262317539. grönwall o., pehrson å. 1984. nutrient content in fungi as a primary food of the red squirrel sciurus vulgaris l. oecologia 64: 230–231. doi:10.1007/bf00376875. kataržytė m., kutorga e. 2011. small mammal mycophagy in hemiboreal forest communities of lithuania. central european journal of biology 6: 446–456. doi:10.2478/s11535-011-0006-z. maser c. a., claridge a.w., trappe j. m. 2008. trees, truffles, and beasts: how forests function. rutgers university press. paolocci f., rubini a., riccioni c., arcioni s. 2006. reevaluation of the life cycle of tuber magnatum. applied and environmental microbiology 72: 2390–2393. doi:10.1128/aem.72.4.2390-2393.2006. pigott c. d. 1985. selective damage to tree-seedlings by bank voles (clethrionomys glareolus). oecologia 67: 367–371. doi:10.1007/bf00384942. small mammal mycophagy 143 riccioni c., belfiori b., rubini a., passeri v., arcioni s., paolocci f. 2008. tuber melanosporum outcrosses: analysis of the genetic diversity within and among its natural populations under this new scenario. new phytologist 180: 466–478. doi:10.1111/j.1469-8137.2008.02560.x. schickmann s., kräutler k., kohl g., nopp-mayr u., krisai-greilhuber i. hackländer k., urban a. 2011. comparison of extraction methods applicable to fungal spores in faecal samples from small mammals. sydowia 63: 237–247. schickmann s., urban a., kräutler k., nopp-mayr u., hackländer k. 2012. the interrelationship of mycophagous small mammals and ectomycorrhizal fungi in primeval disturbed and managed central european mountainous forests. oecologia 170: 395–409. doi:10.1007/s00442-012-2303-2. urban a., neuner-plattner i., krisai-greilhuber i., haselwandter k. 2004. molecular studies on terricolous microfungi reveal novel anamorphs of two tuber species. mycological research 108: 749–758. vernes k., poirier n. 2007. use of a robin’s nest as a cache site for truffles by a red squirrel. northeastern naturalist 14: 145–149. doi:10.1656/1092-6194(2007)14[145:uoarna]2.0.co;2. 2014-01-02t12:10:04+0100 polish botanical society antifungal activity of sodium chloride on saprolegnia diclina and aphanomyces sp. esam h. ali botany department, faculty of science, assiut university assiut, egypt, ibraheem55@yahoo.com ali e. h.: antifungal activity of sodium chloride on saprolegnia diclina and aphanomyces sp. acta mycol. 44 (1): 125–138, 2009. sixteen identified and three unidentified species belonging to six genera of zoosporic fungi were isolated from forty water samples which were collected from different fish and fish hatcheries farms at abbassa city, sharkiya governorate, egypt, using sesame seeds baiting technique at 20±2°c. saprolegnia and achlya contributed the broadest spectra of species diversity amongst the other genera of zoosporic fungi. saprolegnia diclina and aphanomyces sp. were the most prevalent species of zoosporic fungi. the abundance of zoosporic fungal species in these aquacultures was correlated with some physicochemical characteristics of the water samples. the two dominant species of zoosporic fungi were tested for their tolerance of nacl solution and its impact on some morphological and metabolic activities of these fungi. saprolegnia diclina tolerated concentrations of nacl solution till 12000 μg/ml whereas the maximum resistance of aphanomyces sp. was 8000 μg/ml. the examined morphological aspects of the two studied fungal species, which included the colony diameters, the vegetative hyphae, zoosporogenesis, zoospores discharge, sexual reproductive structures and gemmae formation, were generally affected depending upon the tested fungal species and the applied dose of nacl solution. the low treatments of nacl solution were significantly stimulative compared with the control for protease production by s. diclina but higher doses were significantly suppressive. a significant decline in protease activity at all applications was found when aphanomyces sp. was treated with nacl solution. the total free amino acids and total protein content of s. diclina and aphanomyces sp. mycelia were almost significantly increased relative to untreated controls at the low dose of nacl solution and they were significantly dropped at the higher concentrations by the two zoosporic fungi. key words: zoosporic fungi, salinity, morphogenesis and biochemical activities, sporulation acta mycologica vol. 44 (1): 125–138 2009 126 e. h. ali introduction fish diseases constitute one of the most important problems and challenges confronting fish culturists. aquaculture now represents more than 30 percent of total fish production for consumption (delgado et al. 2003). the majority of global production comes from freshwater aquaculture (58%), followed by marine culture (36%) and brackish water (6%). over the past ten years, aquaculture production has increased on average by 11% per year. consequently it is the fastest growing sector of the world food economy (van west 2006). there are a steady increase in fish farms in egypt during the recent years as an alternative protein and a trial to solve the increase in meat prices. outbreaks of waterborne fungal infections (saprolegniasis) on fish and fish eggs continue to cause problems among cultured fish. oomycetes of the order saprolegniales, such as saprolegnia, achlya and aphanomyces species are endemic to all fresh water habitats around the world and they cause the saprolegniasis fish diseases. they are responsible for main types of fungal infections on fish and shellfish in aquaculture, fish farms and hobby fish tanks (neish, hughes 1980; willoughby, pickering 1977; daugherty et al. 1998; bruno, wood 1999; hussein, hatai 2002). oomycetes infections are second only to bacterial diseases in their impact and cause considerable economic losses in aquaculture (meyer 1991; bly et al. 1992; bruno, wood 1999). disease also reduces hatchery efficiency and production, which in turn, increases costs and reduces profit. antifungal agents are essential for the maintenance of healthy stocks of fish and their eggs in intensive aquaculture operations. sodium chloride has been recognized as a safe treatment for saprolegniosis in salmonid incubation and rearing (schreck et al. 1990, 1992; waterstrat, marking 1995). this investigation devoted for studying the occurrence of zoosporic fungi in fish and fish hatcheries farms at abbassa city, el-sharqyia governorate in egypt. the efficacy of nacl as safe antifungal agent on the most prevalent species of zoosporic fungi in these aquacultures was evaluated. in addition, the morphological deviations and some biochemical activities of these zoosporic fungal species as affected by nacl solution were taken into consideration during this study. materials and methods sampling sites and collection of water samples from aquacultures. forty sites were chosen for this study during march 2007 where each small or big water basin was considered an aquaculture site. the forty investigated aquacultures located at abbassa city, sharkiya governorate, egypt and they included both fish farms and fish hatcheries farms. one surface water sample was collected from each aquaculture (fish farm or fish hatcheries farm). each water sample was collected in five brown and sterilized glass bottles (400 ml capacity each). four of these bottles were used for baiting and recovery of zoosporic fungi from waters of aquacultures; where each bottle contained eight germinated and sterilized sesame seeds as capture material antifungal activity 127 of zoosporic fungi. these bottles were aerated at different intervals. the fifth bottle was kept without addition of sesame seeds for the determination of physico-chemical characteristics of aquacultures under investigation. physico-chemical analysis of water samples of aquacultures. physical characteristics of water samples of both fish farms and fish hatcheries included the measurements of water temperature (ranged between 17-21°c) and ph in situ (between 7.85 and 11.03). glass bottles containing water samples free of sesame seeds were used for quantification of total soluble salts and organic matter according to jackson (1958). isolation of zoosporic fungi from fish and fish hatcheries farms (aquacultures). previously prepared glass bottles containing germinated and sterilized sesame seeds were functioned and used for capture and recovery of the resident zoosporic fungi (khallil 1984). water samples in these bottles were poured under aseptic conditions into equivalent numbers of sterilized petri-dishes (15 cm in diameter each). petridishes were kept at 22°c during 24 hours for the colonization of sesame seeds by fungal propagules. colonized sesame seeds were then transferred into other clean and sterile, but smaller, petri-dishes (10 cm in diameter each) containing sterile distilled water to which crystalline penicillin (2000 units per liter of water) was added as indicated by roberts (1963). then, these dishes were incubated at 22°c for twenty days during which they were daily examined, and the recovered zoosporic taxa were identified. most of the recovered zoosporic fungi were purified under aseptic conditions on glucose peptone agar medium (gp) as described by willoughby and pickering (1977). some species of zoosporic fungi required specific media for isolation and purification. allomyces species were purified on yeast peptone starch agar medium ypss (emerson 1941) whilst aphanomyces species were purified on yp agar medium (hatai, egusa 1979). for assessing the total counts of the genera and species which were identified during this investigation, the fungal species appeared on one sesame seed was counted as one colony (isolate) relative to the seed numbers (24/ water sample). identification of the recoverable taxa zoosporic fungi. isolated zoosporic fungi were identified based on morphological characteristics according to coker (1923), johnson (1956, 1971), sparrow (1960), scott (1961), karling (1977), seymour (1970), rattan et al. (1978). selected species of zoosporic fungi and stress solution. two species of zoosporic fungi namely saprolegnia diclina laf47 and aphanomyces sp. laf38, which dominated in fish farms and fish hatcheries were chosen for further morphological and biochemical studies. these species of zoosporic fungi were preserved in the herbarium of our laboratory of aquatic fungi in pure cultures as water sesame seeds cultures and also maintained on slants at 4°c where they renewed every one-two months period. purified nacl was used as stress solution and tested to control the two selected fungal species at different treatments. nacl solution was prepared under aseptic conditions to give different concentrations. these concentrations were 0.0 (control), 2000, 4000, 6000, 8000, 10000 and 12000 μg/ml of nacl solution. media. water sesame seeds cultures (khallil 1984) were used for studying the impact of sodium chloride on the morphological aspects of saprolegnia diclina and aphanomyces sp. glucose peptone (gp) agar medium (willoughby, pickering 1977) was used for purification of saprolegnia diclina and in a broth form (without agar 128 e. h. ali addition) for estimation of the biochemical activities of the species as affected by the different treatments of nacl solution. in case of aphanomyces sp. glucose yeast extract (gy) agar medium (hatai, egusa 1979) was functioned either in a solid form for purification of the species or in a broth form for studying its biochemical activities under salinity (nacl solution) stress. glucose peptone (gp) and gy agar media impregnated with previously mentioned nacl treatments were also used to determine the colony diameters as affected by the different salinity levels. morphological studies under salinity stress. agar discs were excised at the periphery of actively growing mycelia of the two tested species of zoosporic fungi and transferred to petri plates containing sterile distilled water supplemented with germinating and sterilized sesame seeds for the preparation of zoospores suspension. sterilized petri-dishes (12 cm in diameter) containing different concentrations of nacl were enriched under aseptic conditions with sterilized germinating sesame seeds and 2 ml of encysted zoospores suspension of saprolegnia diclina and aphanomyces sp. the experiment was designed in triplicate for each fungal species at each treatment. petri-dishes were then placed in an incubator at 22°c for two weeks during which they were daily examined starting from the second day of incubation using a light olympus microscope provided with camera. the morphological changes in s. diclina and aphanomyces sp. deviated away from the normal shapes were monitored, recorded and were almost photographed. these morphological features included the vegetative hyphae, zoosporangial formation and discharge and sexual reproductive structures. the diameters of the vegetative colonies of the two zoosporic fungal species were measured (cm) by the end of experiment (15 days) at each treatment of nacl solution including controls. average numbers of zoosporangia, sporangial discharge and oogonia were assessed and counted per one colony (sesame seed). biochemical activities under salinity stress. treatments, inoculation and incubation. glucose peptone (gp) and glucose yeast extract (gy) broth media were dispersed into 100 ml erlenmeyer conical flasks (20 ml each) and flasks were plugged and sterilized in an autoclave. then, solutions within flasks were cooled until 40°c and they were adjusted to final concentrations of nacl solution; 0.0 (control), 2000, 4000, 6000, 8000, 10000 and 12000 μg/ml. conical flasks contained gp broth were inoculated under aseptic conditions with 8-d-old gp agar mycelial discs (1 cm) of saprolegnia diclina whereas that contained gy broth were inoculated with 8-d-old gy agar mycelial discs of aphanomyces sp. thereafter, the conical flasks were incubated in an incubator at 22°c for two weeks. the mycelial mats were harvested by vacuum filtration at the end of the incubation period. cultures filtrates were used for the estimation of protease production by the two zoosporic fungal species. the mycelia were washed three times with distilled water and then dried in oven at 75°c until constant weight. dried mycelia were used for the determination of total free amino acids and total protein content. screening for protease activity by test tubes assay. the two species of zoosporic fungi were screened for their activity to secrete protease enzyme on solid medium impregnated with nacl concentrations used in this study. the medium described by paterson and bridge (1994) was employed. the medium was mixed well to obtain miscible medium solution and autoclaved. then, 10 ml volume of the medium were poured under aseptic conditions in test tubes which were kept vertically. the test tubes antifungal activity 129 were inoculated after the medium solidification with 0.5 agar discs taken from the periphery of the actively growing mycelia of each species of the two zoosporic fungi. tri-replicates of test tubes were prepared at each treatment of nacl. test tubes were incubated at 22°c for two weeks after which the depth of clear zone (protease detection) starting from the surface fungal growth was measured and recorded in cm. estimation of protease production. for the determination of protease production, the method described by kunitz (1947) was employed. the quantity of protease were calculated as mg casein/h × 20 ml culture filtrate. total free amino acids. the method of lee and takahashi (1966) was adopted for the determination of total free amino acids of mycelia. the data of amino acids were expressed as mg glycine/g dry weight of mycelia. total protein content. the total protein content was determined using the method of lowry et al. (1951). bovine serum albumin was used as a standard substance and the extinction was measured against appropriate blank at 700 nm. the data obtained were calculated as mg bovine serum albumin/g dry weight of mycelia. statistical analysis. one-way analysis of variance (anova) was used to test effects of nacl concentration on the biochemical activities of s. diclina and aphanomyces sp. using pc stat computer programme. the means at each concentration were compared with control values using lsd test at 5% significant level. results the physico-chemical characteristics of water samples which were collected from the fish and fish hatcheries farms were evaluated. the ph values of aquacultures water samples ranged from 7.85 to 11.03. the total soluble salts of the water samples were relatively low and varied from 6.4 to 53.76 mg/l. the organic matter contents of water samples fluctuated between 189.43 and 497.10 mg/l. the results indicated in table 1 show that sixteen identified in addition to three unidentified species appertaining to six genera of zoosporic fungi were isolated. most of the isolated species during this study are known as fish pathogenic fungi. achlya and saprolegnia contributed the broadest spectra of species diversity (five species each) recovered from the investigated aquacultures. the most dominant species of zoosporic fungi were saprolegnia diclina and aphanomyces spp (highly occurred). the species of achlya dubia, dictyuchus monosporus and pythium rostratum were considered as of moderate occurrence in these aquacultures. the rest of species of zoosporic fungi were either isolated in low or rare occurrence. saprolegnia was the dominant genus in the surface waters of fish and fish hatcheries farms and it was of high occurrence (32 out of 40 water samples). it was represented by five species and thus ranking with achlya the broadest spectra of the isolated species. of the isolated species; s. diclina was the most prevalent (highly occurred; 24 water samples) inhabiting waters of fish and fish hatcheries farms in the studied area. the remaining species were s. ferax (low incidence; 6 water samples), s. furcata, s. glomerata and the unidentified species (rare occurrence; 2-4 water samples). aphanomyces came next after saprolegnia and it was also of high occurrence (26 130 e. h. ali water samples). this genus included three species of which the unidentified species (high incidence; 21 samples) were the most prevalent in the waters of aquacultures. the other two species namely; a. scaber and a. laevis were isolated in low to rare occurrence (6 and 2 water samples, respectively). achlya was also of high occurrence and it contributed five species of which a. dubia was the commonest (moderately occurred; 12 water samples). the other species namely; a. debaryana and a. racemosa which were of low incidence (6 and 5 water samples) whilst a. diffusa and the unidentified species were of rare occurrence (2 samples). dictyuchus was of moderate occurrence (16 water samples) and it was represented by two species namely d. monosporus (moderately occurred; 11 samples) and d. sterilis (low incidence; 5 water samples). pythium was also moderately encountered from 15 water samples of aquacultures and it was included three species of which p. rostratum moderately prevailed (12 samples). pythium debaryanum and p. ultimum regarded as of rare occurrence (2-3 water samples). allomyces was of rare incidence (4 samples) in waters of aquacultures and it was represented by only one species (a. macrogynus). morphological studies. the data presented in table 2 show that the diameters of the vegetative colonies of both s. diclina and aphanomyces sp. on the solid media were generally decreased with uprising the concentration of nacl solution. the radial growth of s. diclina was greater than that of aphanomyces sp. at controls and the parallel concentrations of nacl solution. table 1 total counts, cases of isolations and occurrence remarks of zoosporic fungi collected from fish and fish hatcheries farms genera and species of zoosporic fungi number of isolates cases of isolations occurrence remarks achlya a. diffusa harvey ex johnson a. debaryana humphrey a. dubia coker a. racemosa hildebrand achlya sp. allomyces macrogynus emerson & wilson aphanomyces aphanomyces laevis de bary a. scaber de bary aphanomyces sp. dictyuchus d. monosporus leitgeb d. sterilis coker saprolegnia s. diclina humphrey s. ferax (gruith.) thuret s. furcata maurizio s. glomerata (tiesenhausen) lund saprolegnia sp. pythium p. debaryanum hesse p. rostratum butler p. ultimum trow 156 19 37 72 20 8 13 115 6 17 92 103 58 45 257 162 45 21 10 18 60 10 43 7 23 2 6 12 5 2 4 26 2 6 21 16 11 5 32 24 6 4 2 2 15 3 12 2 h r l m l r r h r l h m m l h h l r r r m r m r total number of isolates 704 abbreviations: h – high occurrence; more than 20 water samples; m – moderate occurrence between 10-20 water samples; l – low occurrence; between 5-9 water samples; r – rare occurrence less than 5 samples. antifungal activity 131 saprolegnia diclina treated with 2000 μg/ml of nacl solution show that both sporangial formation and discharge were little affected compared with that at controls (fig. 1) and they were still in high numbers. sexual reproductive structures (oogonia and antheridia) were of moderate numbers. compared with the oogonia at controls (fig. 2), the oogonia at this treatment expanded in their sizes and oospheres contents. the gemmae were of high number and were similar in shape to controls (fig. 3). at 4000 μg/ml of nacl solution, zoosporogenesis (sporangial and zoospores formation) was observed in moderated number whereas discharge was remarked in low number. low number of oogonia and antheridia were observed at this concentration and oogonia showed mild symptoms of plasmolysis. the resting bodies of gemmae appeared in high numbers. at 6000 μg/ml of nacl solution, zoosporogenesis occurred in low number and zoospores discharge rarely recorded. rare number of plasmolysed oogonia (fig. 4) and non-functional antheridia were shown at this treatment of nacl solution. the resting structures (gemmae) were remarked in moderate number and expanded in their shapes (fig. 4). at 8000 μg/ml of nacl the vegetative hyphae were loosed in their appearance. zoosporangia were observed in rare number and they displayed neither sporulation nor spores liberation. rudimental oogonia (fig. 5) were formed in rare numbers and antheridia were non-functional. gemmae were also represented in rare numbers. at the application of 10000 μg/ml of nacl solution the vegetative hyphae were thin compared with the control. zoosporangia were of rare number and showed unusual small appendages appearance (fig. 6). these sporangia never showed spores differentiation and liberation. the sexual apparatus was represented by non-functional filamentous antheridia and gemmae were of rare numbers. the maximum tolerance (sub-lethal concentration) of s. diclina for nacl solution was 12000 μg/ml. at this treatment, the vegetatable 2 effects of nacl solution on some morphological characteristics of saprolegnia diclina and aphanomyces sp. zoosporic fungi nacl conc. μg/ml morphological characteristics colonies diameter (cm) sporangia formation sporangia discharge oogonia and antheridia gemmae formation s. diclina 0.0 2000 4000 6000 8000 10000 12000 3.5 3.2 3.0 2.7 2.3 1.6 0.9 31 h 27 h 12 m 5 l 2 r 2 r 31 h 27 h 5 l 2 r 26 h 11 m 5 l 2 r 2 r na na 34 h 32 h 27 h 18 m 4 r 3 r aphanomyces sp. 0.0 2000 4000 6000 8000 1.1 1.0 0.8 0.6 0.5 40 h 43 h 18 m 4 r 40 h 43 h 15 m 2 r abbreviations: h – high number per one colony; more than 14 sporangia, > 12 oogonia, > 21 gemmae in case of s. diclina and > 20 sporangia for aphanomyces sp.; m – moderate number per colony; 7-14 sporangia, 6-12 oogonia, 10-21 gemmae in case of s. diclina and 10-20 sporangia for aphanomyces sp.; l – low number per colony; 3-5 sporangia, 3-6 oogonia, 5-9 gemmae in case of s. diclina and 5-9 sporangia for aphanomyces sp.; r – rare number per colony; less than 3 sporangia, < 3 oogonia, < 5 gemmae in case of s. diclina and < 5 sporangia for aphanomyces sp.; na – means only non-functional antheridia appeared; – means structures did not appear. 132 e. h. ali tive hyphae (thin) showed only non-functional antheridial appendages and neither sporangia or sporangial discharge were observed. aphanomyces sp. was sensitive for nacl solution treatment in comparison with s. diclina where its maximum tolerance was 8000 μg/ml. the vegetative hyphae of aphanomyces sp. were of sparse lateral branching (fig. 7) at controls and they were of condensed growth and profusely branched (fig. 8) at 2000 μg/ml applications of nacl solution. this dose was stimulative for zoosporogenesis and zoospores liberations (high numbers). at 4000 μg/ml of nacl solution, the vegetative hyphae showed minor lateral short branches formation compared with the control. sporangial formation as well as zoospores release were moderately observed. the vegetative hyphae at 6000 μg/ml of nacl solution seemed loose and spiral in their structure and zoosporangial formation and discharges were of rare numbers. the sub-lethal treatment of nacl solution was 8000 μg/ml at which only sterile and stunted vegetative hyphae appeared. biochemical studies. the results shown in table 3 reveal that, the total free amino acids content of s. diclina significantly raised compared with the control at the lowest application of nacl solution (2000 μg/ml) and it slightly dropped (non-significantly) at 4000 μg/ml. with increasing nacl solution until the sublethal treatment (12000 μg/ml), the mycelial amino acids of s. diclina were significantly decreased relative to untreated controls. in instance of aphanomyces sp., the total free amino acids of the mycelia were increased non-significantly comparable to the control at the low supplement of nacl solution (2000 μg/ml) and they relatively declined at 4000 μg/ml. the highest two applications of nacl solution (6000 and 8000 μg/ml) were inhibitory (significantly) for the mycelial content of amino acids. the data presented in table 3 indicate that, the total protein content of s. diclina mycelia was significantly promoted at the low treatment of nacl solution (2000 μg/ ml). all the other tested concentrations (4000-12000 μg/ml) were found inhibitory for the mycelial content of protein which was decreased with rising the concentration table 3 effect of different treatments of nacl on total free amino acids (taa, mg glycine/g dry weight of mycelia), total protein content (tp, mg bovine albumin/g dry weight of mycelia), and protease detection (pd; cm) and production (pp, μg bovine albumin/20 ml culture medium) by s. diclina and aphanomyces sp. zoosporic fungal species concs. (μg/ml) biochemical activities protease detection (pd; cm) taa tp pp s. diclina 0.0 2000 4000 6000 8000 10000 12000 0.41 0.47* 0.39 0.22* 0.18* 0.17* 0.15* 34.01 41.22* 31.01* 24.84* 20.70* 16.69* 13.18* 12.82 14.85* 14.12* 7.41* 6.28* 5.71* 4.91* 6.4 5.7 4.6* 1.8* 1.2* 0.5* lsd at 5% level 0.04 1.20 1.29 1.3 aphanomyces sp. 0.0 2000 4000 6000 8000 0.44 0.45 0.39 0.17* 0.14* 21.48 22.20 20.06* 16.43* 12.96* 18.36 15.23 11.01* 8.00* 7.12* 6.7 4.1* 1.6* 0.6* lsd at 5% level 0.09 0.77 3.27 1.5 abbreviations: lsd – least significant difference; * significant difference compared with the control; protease enzyme was not detected at this treatment. antifungal activity 133 of nacl solution. regarding aphanomyces sp., the total protein content of the mycelia activated (non-significantly) only comparable with the control at the low dose of nacl solutions (2000 μg/ml) and the other applied concentrations (4000-8000 μg/ml) significantly suppressed the protein content. as presented in table 3, protease production by s. diclina was induced significantly at 2000 and 4000 μg/ml of nacl solution. thereafter, as the concentration of nacl solution increased, the protease activity of s. diclina significantly suppressed comparable with the control. in case of aphanomyces sp., protease activity was almost significantly inhibited at all the tested concentrations of nacl solution. the quantity of protease enzyme by aphanomyces sp. was greater than that of s. diclina at controls and at 2000, 6000 and 8000 μg/ml of nacl solution. the data presented in table 3 show that protease enzyme detection, as indicated by clear zone (fig. 9), on solid medium was inhibited significantly at most different nacl concentrations compared with the control by the two tested fungal species. no protease activity was detected at the sublethal dose of nacl solution of each fungal species. discussion the ph values of the water samples were alkaline between 7.85 and 11.03 and they had no role on the occurrence of species of zoosporic fungi in the investigated aquacultures. the total soluble salts of aquacultures water samples were found low (6.4 to 53.76 mg/l) and they had no effect on the incidence of the isolated fungal species. the organic matter contents of aquacultures water samples fluctuated between 189.43 and 497.10 mg/l and the waters had high content of organic matter were the richest in the isolated species of zoosporic fungi. more or less similar results were obtained by kiziewicz et al. (2004) who found that physicochemical parameters of waters had no important effect on the occurrence of zoosporic fungi isolated from bathing sites of the suprasl river. in addition, jeffrey et al. (1985) the production of viable zoospores (zoosporogenesis) by lagenidium giganteum took place from ph 4.5 to 8.4 by three isolates and from 4.5 to 8 by two other isolates (±0.2). also, piotrowski et al. (2004) reported that isolates of the chytrid betrachochytrium dendrobatidis grew and reproduced at ph 4-8 and the growth was maximal at ph 6-7. most of the recoverable species of zoosporic fungi (sixteen identified and three unidentified species) during this study are economically important as fish pathogenic fungi. it was found that saprolegnia diclina and aphanomyces spp were the highly occurred zoosporic fungi in the investigated aquacultures. the species of achlya dubia, dictyuchus monosporus and pythium rostratum were also common but they were of less extent. the other isolated species were recovered either in low or rare occurrence. in this regards, ogbonna and alabi (1991) isolated 24 zoosporic fungal species from the infected fish in a nigerian freshwater fish pond. achlya racemosa, aphanomyces laevis, dictyuchus sterilis, saprolegnia ferax, s. litoralis and s. parasitica had 100% frequency of occurrence. there were similarities in the species of fungi 134 e. h. ali isolated from the infected fishes in the fish pond and those isolated from the hatchery. also, kiziewicz et al. (2004) identified thirty six fungi species in the bathing sites of the suprasl river, among them fish pathogens achlya orion, aphanomyces laevis, dictyuchus monosporus, saprolegnia ferax, saprolegnia monoica and s. parasitica. in addition, khulbe et al. (1994) reported achlya debaryana (saprolegniales, oomycetes) for the first time as a fish pathogen in a huge artificial reservoir and recognized fish production center in naini tal district, india. the two tested fish pathogenic fungi varied in their tolerance rates of nacl solution. saprolegnia diclina tolerated against nacl solution until 12000 μg/ml μg/ml whilst aphanomyces sp. could resist only until 8000 μg/ml of nacl solution. the diameters of the vegetative colonies of the tested oomycetous fungi decreased with increasing the dose concentration of nacl solution. in this respect, several authors found that nacl reduced growth and was the most effective in controlling of phytophthora cinnamomi (sterne et al. 1976), paecilomyces lilacinum and aspergillus niger (mert, dizbay 1977), saprolegnia diclina (taylor, bailey 1979), saprolegnia spp. (martinez-palacios et al. 2004; khodabandeh, abtahi 2006) and saprolegnia parasitica (ali 2005). however, mert and ekmekci (1987) reported that the vegetative growth of aspergillus flavus and penicillium chrysogenum increased with an increase in the nacl content of the nutrient medium. the vegetative hyphae of s. diclina appeared loose and thinner at the high concentrations of nacl solution whereas that of aphanomyces sp. were profusely branched compared with the control at the low applications and they displayed spiral at the high supplements. in accordance with these results, katz and rosenberg (1971) observed that numerous septa and branches are formed when aspergillus nidulans is exposed to osmotic shock. more or less similar result was obtained by ali (2005). zoosporogenesis and zoospores discharge in s. diclina were little affected at the low treatment of nacl solution. they were dropped in numbers and deformed in shapes with increasing the dose concentration of nacl solution while they switched off at the sublethal dose in case of the two tested species of zoosporic fungi. the low dose of nacl solution was found to be inducible for zoosporogenesis and zoospores discharge in case of aphanomyces sp. in agreement with these findings, some investigators found that relatively low concentrations of nacl suppressed or eliminated zoosporogenesis and reproductive growth in trichophyton mentagrophytes (kane, fischer 1973), saprolegnia species (harrison, jones 1975; smith et al. 1990), lagenidium giganteum (jeffrey et al. 1985), aspergillus flavus (mert, ekmekci 1987) and saprolegnia parasitica (ali 2005). kirvu et al. (2005) characterized sporulation and cyst formation of secondary zoospores of two isolates of a. invadans which prevalent in atlantic menhaden, brevoortia tyrannus at different salinities and found that sporulation occurred only at low salinities. from laboratory observations, they concluded that low salinities appeared crucial to transmission of the pathogen. the two isolates of a. invadans produced free-swimming secondary zoospores at salinities of 0, 1 and 2 psu (salinity unit = per thousand), but not at 4 psu or higher. sporangial formation and zoospores release by the oomycete fish pathogen the reproductive structures, oogonia and antheridia, produced by s. diclina responded variably and depended upon the dose concentration of nacl solution. oogonia declined in numbers but were enlarged in size and increased in the number of oospheres content at the low application of nacl. they showed subsequent decrease antifungal activity 135 in their numbers and morphological alterations ranged between mild and severe plasmolysis, rudimental structure and appeared without their antheridial attach with rising the concentration of nacl solution. sexual structures were represented by only non-functional antheridia at the sublethal application of nacl solution. in confirmation with these results, kane and fischer (1973) reported that the addition of 3–5% nacl to the incubation medium inhibited the reproductive growth of trichophyton mentagrophytes. moreover, harrison and jones (1975) and smith et al. (1990) claimed that oogonium production of pathogenic and non-pathogenic saprolegnia species is suppressed by relatively low salt concentrations. the gemmae bodies formation by s. diclina was activated at the low concentrations of nacl solution where they were recorded abundant. it may attributed for unsuitable conditions being resulted from the increase of salinity level to tolerate them. in this regards, faye et al. (2006) reported that sclerotial yield and sclerotial germination of five isolates of rhizoctonia solani declined with decreasing osmotic potential on osmotically adjusted media. however, higher concentrations of nacl solution were inhibitory for gemmae formation in s. diclina compared with that at controls. the s. diclina and aphanomyces sp. produced extracellular protease enzyme in the culture filtrates at control and treated cultures. the quantity of protease secretion varied between the two tested fungi where aphanomyces sp. was nearly more potent than s. diclina in protease production at the parallel concentrations of nacl solution. protease screening by the two tested species of zoosporic fungi on solid medium showed a measurable reduction as the concentrations of nacl increased. the ability of zoosporic fungi to produce protease was followed by weiland (2004) who examined the production of protease by the sugarbeet pathogen aphanomyces cochlioides, the legume pathogen a. euteiches, and the fish pathogen saprolegnia parasitica. protease activity was readily detected in supernatants of water cultures of each organism using autoclaved host tissue as a nutrient source. for the three oomycetes tested, abundant protease activity was present in culture supernatants; additionally, high protease levels were detected in host seedlings infected with the two phytopathogens. in inoculated sugarbeet seedlings, the protease activities were detected prior to or concomitant with the onset of disease symptoms and the activities were capable of digesting protein extracted from sugarbeet hypocotyls. also, soederhaell and unestam (1975) found protease activity in culture filtrates from the crayfish plague parasite, aphanomyces astaci. in addition, dieguez-uribeondo and cerenius (1988) reported that species of aphanomyces that attack fish (e.g., a. invidans) and crayfish (e.g., a. astaci) have been shown to produce abundant extracellular protease. the penetration of the crayfish cuticle by aphanomyces astaci (bangyeekhun et al. 2001) and of human epidermis by pythium insidiosum (ravishankar et al. 2001) is proposed to involve digestion by protease. however, leger et al. (1986) found that several pathogenic isolates of metarhizium anisopliae, beauveria bassiana, and verticillium lecanii when grown in buffered liquid cultures containing comminuted locust cuticle as composite carbon source, produced a variety of extracellular enzymes corresponding to the major components of insect cuticle which included proteases. considerable variations occurred in levels of production between species and even within a species, but proteases were exceptional as production of them was high with all the isolates. the low supplements of nacl solution (2000 and 4000 μg/ml) were significantly stimulative for protease production by s. diclina compared with the control whereas 136 e. h. ali the higher concentration were significantly suppressive for the enzyme production. however, protease production by aphanomyces sp., was almost significantly dropped at all doses of nacl solution. the elevated concentrations of nacl solution (starting from 6000 μg/ml) significantly dropped the mycelial content of amino acids of the two tested species of zoosporic fungi comparable to untreated controls. similar result was also obtained by ali (2005). on the other hand, wethered and jenning (1985) found that amino acids within the mycelia of marine fungi increased with increasing salinity. the total protein contents of s. diclina and aphanomyces sp. mycelia were significantly decreased at the tolerant concentrations of nacl solution (except at the low dose) as compared with the control. in agreement with this result, ali (2005) found that the total protein content of saprolegnia parasitica was inhibited at all tested concentrations of nacl solution. conclusions several species of the isolated zoosporic fungi from the study zone are known as fish pathogenic fungi. this information gives a sign for possible infection of the cultured fish species in the investigated fish and fish hatcheries farms by these fungi. as a result problems can face culturists and investments in these aquacultures. the results of testing the effect of nacl solution on the most two dominant zoosporic fungal species (known as fish pathogens) in these farms indicated that the high salinity levels were successful in controlling and suppressing the activities of these fungi. references ali e. h. 2005. morphological and biochemical alterations of oomycete fish pathogen saprolegnia parasitica as affected by salinity, ascorbic acid and their synergistic action. mycopathologia 59: 231–243. bangyeekhun e., cerenius l., soderhall k. 2001. molecular cloning and characterization of two serine proteinase genes from the crayfish plague fungus, aphanomyces astaci. j. invertebr. pathol. 77: 206–216. bly j.e., lawson l.a., dale d.j., szalai a.j., durborow r.m., clem l.w. 1992. winter saprolegniosis in channel catfish. dis. aqu. organ. 13: 155–164. bruno d.w., wood b.p. 1999. saprolegnia and other oomycetes. 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grzywacz 1995; kowalski 2001; stocki 2001; przybył 2002; vasiliauskas et al. 2006), the disease spread consequently to reach ashes in scandinavia, western europe, and some regions of southern europe (kirisits et al. 2009; ioos et al. 2009; timmermann et al. 2011; hauptman et al. 2012). in 2012 the ash dieback symptoms were also reported from british isles (coghlan 2012). the fungus chalara fraxinea t. kowalski is believed to be the main causal agent of ongoing ash dieback process. this species was originally described based on cultures isolated from necrotic areas on ash shoots collected in włoszczowa forest district (kowalski 2006, 2007). the artificial inoculation of acta mycologica vol. 48 (2): 135–146 2013 doi: 10.5586/am.2013.031 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 136 t. kowalski et al. ash shoots proved its high pathogenicity (bakys et al. 2009; kowalski, holdenrieder 2009a). the occurrence of chalara fraxinea was confirmed in more than twenty european countries where the ash dieback symptoms were observed (halmschlager, kirisits 2008; ioos et al. 2009; timmermann et al. 2011). further studies revealed that chalara fraxinea is an anamorphic state of fungus from the genus hymeno­ scyphus. it was initially believed that it was hymenoscyphus albidus (robergere ex desm.) w. phillips, known in europe since 1851 (kowalski, holdenrieder 2009b). this opinion was however changed on the basis of molecular analyses which showed that this species is comprised of two, genetically distinct, subpopulations. they are hardly distinguishable morphologically, but they differ significantly in their potential to generate disease symptoms. eventually the highly virulent subpopulation of h. albidus was recognized to be the teleomorphic state of chalara fraxinea and described as a new cryptic species hymenoscyphus pseudoalbidus queloz et al. (queloz et al. 2011). it produce the apothecia on previous year ash leaf rachises in the stand floor. the molecular studies based on two-hundred and thirty apotheciaderived cultures originating from four regions of poland, confirmed the presence of highly virulent h. pseudoalbidus in declining ash stands while h. albidus was not found (kraj, kowalski 2013). the apothecia of h. pseudoalbidus are produced mainly from early july to the end of september (kowalski 2012). this is also the season when apothecia release the ascospores that infect ashes and initiate their disease process. the number of h. pseudoalbidus apothecia produced in the stand floor indicates the amount of infectious material and consequently the risk level for fraxinus excelsior stands. the primary goal of the study was the estimation of the overall number of h. pseudoalbidus apothecia that may be produced on ash rachises in the leaf litter in the selected stands in southern poland. additionally, in order to correlate their presence with the stand health, the intensity of the selected disease symptoms for this stands was estimated. materials and methods the study was performed in august 2011 in 14 stands of myślenice and 14 stands of dynów forest districts. the selected stands represented mostly fresh upland broadleaved forest habitat, but they varied in age. the percentage of ash ranged from 40 to 90% for most stands, as only a few of them were characterized by lower proportion of ashes (tabs 1, 2). the stands were selected randomly using information from forest management plans. fifty centrally located neighboring trees of fraxinus excel­ sior were examined for their health condition in each stand. each examined tree was categorized as: i/ dead, ii/ with visible disease symptoms, or iii/ with no macroscopic disease symptoms. furthermore, additional data were recorded for all trees classified as type ii: the presence of local trunk necroses, the presence of dead top, and the crown condition measured by the percentage of dead branches ranked as: below 25%, 26-50%, 51-75% and above 76% (fig. 1). in total, the health condition evaluation was carried out for 1400 trees, 700 per each of two forest districts (tabs 1, 2). 138 t. kowalski et al. ta bl e 1 t he o cc ur re nc e of d is ea se s ym pt om s on f ra xi nu s ex ce ls io r a nd th e oc cu rr en ce o f t he h . p se ud oa lb id us a po th ec ia on a sh le af r ac hi se s in th e lit te r in m yś le ni ce f or es t d is tr ic t a ge cl as s (y ea rs ) c om pa rt m en t su bc om pa rt m en t si te ty pe *1 a sh pe rc en ta ge n um be r of an al yz ed tr ee s n um be r (% ) of tr ee s n um be r of p lo ts n um be r of ra ch is es *3 n um be r of ap ot he ci a* 3 de ad w it h de ad to p w it h de ad br an ch es sy m pt om le ss on pl ot s pe r 1 m 2 on pl ot s pe r 1 m 2 < 2 0 3 g l a 60 50 23 25 2 5 23 18 16 8 13 4 26 f l a 10 50 18 23 9 5 57 46 28 0 22 4 σ 10 0 41 (4 1. 0) 48 (4 8. 0) 11 (1 1. 0) 10 80 32 44 8 17 9 21 -4 0 t p 1* 2 l c 50 50 8 40 2 6 43 29 74 8 49 9 48 b l b 60 50 18 28 4 6 21 14 33 0 22 0 σ 10 0 26 (2 6. 0) 68 (6 8. 0) 6( 6. 0) 12 64 21 10 78 35 9 41 -6 0 9 d l a 40 50 12 35 3 6 92 61 57 1 38 1 29 h l a 50 50 9 37 4 6 10 0 67 99 8 66 5 99 d l c 80 50 6 41 3 6 82 55 89 9 59 9 30 4 a l c 70 50 8 41 1 6 87 58 20 03 13 35 σ 20 0 35 (1 7. 5) 15 4( 77 .0 ) 11 (5 .5 ) 24 36 1 60 44 71 74 4 > 6 0 27 a l c 10 50 30 19 1 6 55 37 69 0 46 0 13 6 d l d 40 50 9 40 1 6 96 64 13 57 90 5 27 9 b l a 60 50 5 2 41 2 6 67 45 11 95 79 7 27 9 c l a 50 50 9 2 38 1 6 91 61 13 30 88 7 27 9 d l a 60 50 1 1 42 6 6 13 4 89 13 05 87 0 28 0 g l a 80 50 4 43 3 6 98 65 60 3 40 2 σ 30 0 58 (1 9. 3) 5( 1. 7) 22 3( 74 .3 ) 14 (4 .7 ) 36 54 1 60 64 80 72 0 to ta l 70 0 16 0( 22 .9 ) 5( 0. 7) 49 3( 70 .4 ) 42 (6 .0 ) 82 10 46 51 12 47 7 60 9 *1 l a = fr es h up la nd d ec id uo us fo re st , l b = w et u pl an d de ci du ou s fo re st , l c = r ip ar ia n up la nd d ec id uo us fo re st , l d = w et m ou nt ai n de ci du ou s fo re st *2 t p 1 – th e st an d es ta bl is he d on fo rm er a gr ic ul tu ra l l an d *3 th e av er ag e va lu es w er e ro un de d to fu ll nu m be r the occurrence of hymenoscyphus pseudoalbidus 139 ta bl e 2 t he o cc ur re nc e of d is ea se s ym pt om s on f ra xi nu s ex ce ls io r a nd th e oc cu rr en ce o f t he h . p se ud oa lb id us a po th ec ia on a sh le af r ac hi se s in th e lit te r in d yn ów f or es t d is tr ic t a ge cl as s (y ea rs ) c om pa rt m en t su bc om pa rt m en t si te ty pe *1 a sh pe rc en ta ge n um be r of an al yz ed tr ee s n um be r (% ) of tr ee s n um be r of p lo ts n um be r of ra ch is es *2 n um be r of ap ot he ci a* 2 de ad w it h de ad to p w it h de ad br an ch es sy m pt om le ss on pl ot s pe r 1 m 2 on pl ot s pe r 1 m 2 < 2 0 57 d l a 50 50 5 45 6 42 28 24 0 16 0 95 b l a 70 50 8 1 41 6 36 24 22 0 14 7 95 c l a 80 50 11 2 37 6 31 21 15 4 10 3 96 c l a 80 50 40 10 6 26 17 16 9 11 3 12 5 n l c 50 50 4 5 41 6 8 5 25 17 σ 25 0 28 (1 1. 2) 8( 3. 2) 20 4( 81 .6 ) 10 (4 .0 ) 30 14 3 19 80 8 10 8 21 -4 0 68 a l a 10 50 1 1 48 6 77 51 46 4 30 9 97 d l c 20 50 15 32 3 6 38 25 28 8 19 2 σ 10 0 1( 1. 0) 16 (1 6. 0) 80 (8 0. 0) 3( 3. 0) 12 11 5 38 75 2 25 0 41 -6 0 98 i l a 30 50 2 46 2 6 89 59 56 1 37 4 10 7 a l c 60 50 3 47 6 40 27 25 8 17 2 10 7 d l a 90 50 3 5 42 6 55 37 46 6 31 1 10 8 a l a 50 50 4 7 39 6 65 43 32 6 21 7 10 9 a l c 50 50 18 32 6 81 54 42 0 28 0 σ 25 0 7( 2. 8) 35 (1 4. 0) 20 6( 82 .4 ) 2( 0. 8) 30 33 0 44 20 31 27 1 > 60 11 0 a l c 80 50 24 26 6 41 27 26 0 17 3 12 5 t l c 10 50 1 6 43 6 10 7 55 37 σ 10 0 1( 1. 0) 30 (3 0. 0) 69 (6 9. 0) 12 51 17 31 5 10 5 to ta l 70 0 37 (5 .3 ) 89 (1 2. 7) 55 9( 79 .9 ) 15 (2 .1 ) 84 68 0 32 41 66 19 8 *1 l a = fr es h up la nd d ec id uo us fo re st , l c = r ip ar ia n up la nd d ec id uo us fo re st *2 th e av er ag e va lu es w er e ro un de d to fu ll nu m be r 142 t. kowalski et al. fig. 5. the hymenoscyphus pseudoalbidus apothecia on previous year ash rachises in the litter (rachises covered by black fungal plectenchyma). fig. 6. fragment of ash leaf rachis with one mature apothecium and several apothecia in the initial stage of development. the occurrence of hymenoscyphus pseudoalbidus 143 discussion the characteristic symptoms of ash dieback were observed in all analyzed stands of myślenice and dynów forest districts. the nature of these symptoms was the same as the ones observed in the other parts of poland (kowalski, czekaj 2010), or in other european countries (ioos et al. 2009; kirisits et al. 2009; hauptman et al. 2012). however, the intensity of disease process, measured by the number of trees with high proportion of dead branches, trees with dead tops, or entirely dead trees varied significantly between two forest districts or even between stands of particular forest districts. these differences result from the age of trees, habitat properties, and the ash susceptibility (gil et al. 2006; jaworski 2011; mckinney et al. 2011; kowalski et al. 2012; stener 2013). local conditions are also an important factor that affects the formation of the infectious material reservoirs of h. pseudoalbidus (anamorph: chalara fraxinea), the ash dieback causal agent (kirisits et al. 2009; holdenrieder 2012; kowalski et al. 2012). according to previous research, the chalara fraxinea conidia cannot germinate and the f. excelsior trees are infected exclusively by wind-borne ascospores, that are produced in apothecia on ash leaf residues in the litter (kirisits et al. 2009; kowalski 2012; gross, holdenrieder 2013). only occasionally apothecia may develop on dead ash shoots (kowalski et al. 2010; holdenrieder 2012). beside shoots, also leaves are infected by ascospores. the h. pseudoalbidus infected rachises of fallen leaves darken due to formation of melanized, stroma-like plectenchyma that functions as protection against competing microorganisms, and against unsuitable environmental conditions (gross, holdenrieder 2013). all h. pseudoalbidus apothecia-carrying rachises, as well as many apothecia-free ones, were covered by plectenchyma. the total numbers of ash rachises on experimental plots of particular analyzed stands were varied. this resulted mainly from the various proportion of ash in the species composition of the stand, but also from the health condition of trees, especially from the level of foliage reduction. on the other hand, the relationship between the total number of rachises on plots and the number of apothecia-carrying rachises may be affected by many factors. the development of apothecia may be delayed or even prevented by excessively dry conditions (holdenrieder 2012; kowalski et al. 2012). furthermore, h. pseudo­ albidus is a heterothalic fungal species, thus the apothecia formation requires the infected rachis to contain both mating types (gross et al. 2012). finally, the proportion of apothecia-carrying rachises depends on the on-tree leaf infection frequency by this fungus. if the leaves do not fall due to disease process but naturally, or the premature leaf falling is the result of the trunkor the basal-branch-part necrosis, the proportion of h. pseudoalbidus infected rachises is lower. the above factors were most probably the reasons why the total number of apothecia counted on 82 plots in myślenice forest districts exceeded threefold their number on 84 plots in dynów forest district. those factors probably affected also the number of apothecia per 1 m2 estimated for various stands, as they varied significantly in both forest districts. these differences are evidenced by data showed in tables 1 and 2. the data, when extrapolated to the area of 1 hectare, show even more explicitly that the amount of infectious material in stands with the presence of f. excelsior is huge. in 144 t. kowalski et al. fact, the number of the apothecia produced in a given stand is even greater, as the apothecia formation on the particular rachis is not a one-off exercise but a continuous process. while some apothecia are fully developed and release ascospores, some others are in initial, hardly visible, stage that can be seen on figure 6. in poland, the h. pseudolabidus apothecia are produced for about 3 months usually from july to september. acknowledgements. the investigations were carried out under the project supported by the national science centre, decision no. dec-2011/03/b/nz9/00078. authors express their gratitude to mgr inż. stanisław widz (head of myślenice forest district) and mgr inż. adam pilch (head of dynów forest district) for the help during the research. references bakys r., vasaitis r., barklund p., thomsen i.m., stenlid j. 2009. occurrence and pathogenicity of fungi in necrotic and non-symptomatic shoots of declining common ash (fraxinus excelsior) in sweden. eur. j. forest res. 128: 51-60. http://dx.doi.org/10.1007%2fs10342-008-0238-2 coghlan a. 2012. are europe’s ash trees finished? retrieved 31.10.2012, from the database at http:// 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(in:) t. doğmuş-lehtijärvi (ed.). proceedings of the conference of iufro working party 7.02.02, eğirdir, turkey, 11-16 may 2009. sdu faculty of forestry journal, issn: 13027085, serial: a, special issue: 97-119. kowalski t. 2001. o zamieraniu jesionów. trybuna leśnika 4, 359: 6-7. kowalski t. 2006. chalara fraxinea sp. nov. associated with dieback of ash (fraxinus excelsior) in poland. for. path. 36: 264-270. http://dx.doi.org/10.1016%2fj.fgb.2012.08.008 kowalski t. 2007. chalara fraxinea – nowo opisany gatunek grzyba na zamierających jesionach w polsce. sylwan 151 (4): 44-48. 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(in:) d. dujesiefken (ed.). jahrbuch der baumpflege, haymarket media, braunschweig: 184-195. kraj w., kowalski t. 2013. genetic variability of hymenoscyphus pseudoalbidus dieback cause of european ash (fraxinus excelsior l.). j. phytopathology http://dx.doi.org/10.1111/jph.12173 mckinney l.v., nielsen l.r., hansen j.k., kjær e.d. 2011. presence of natural genetic resistance in fraxinus excelsior (oleaceae) to chalara fraxinea (ascomycota): an emerging infectious disease. heredity 106: 788-797. http://dx.doi.org/10.1038%2fhdy.2010.119 przybył k. 2002. fungi associated with necrotic apical parts of fraxinus excelsior shoots. for. path. 32: 387–394. http://dx.doi.org/10.1046%2fj.1439-0329.2002.00301.x queloz v., gruenig c.r., berndt r., kowalski t., sieber t.n., holdenrieder o. 2011. cryptic speciation in hymenoscyphus albidus. for. path. 41: 133-142. http://dx.doi.org/10.1111%2fj.14390329.2010.00645.x sierota z., stocka t., małecka m., duda-kiełczewska b., oszako t. 1993. ocena występowania ważniejszych szkodników leśnych i chorób infekcyjnych w polsce w roku 1992 oraz prognoza ich pojawu w roku 1993. instytut badawczy leśnictwa, warszawa, 137 pp. stener l.-g. 2013. clonal differences in susceptibility to the dieback of fraxinus excelsior in southern sweden. scand. j. forest res. 28 (3): 205-216. http://dx.doi.org/10.1080%2f02827581.2012.735699 stocki j. 2001. przyczyny zamierania drzew i drzewostanów jesionowych w polsce. głos lasu 4: 17-19. timmermann v., břrja i., hietala a.m., kirisits t., solheim h. 2011. ash dieback: pathogen spread and diurnal patterns of ascospore dispersal, with special emphasis on norway. bulletin eppo 41: 14-20. vasiliauskas r, bakys r, lygis v, ihrmark k, barklund p, stenlid j. 2006. fungi associated with the decline of fraxinus excelsior in the baltic states and sweden. (in:) t. oszako, s. woodward (eds). possible limitations of dieback phenomena in broadleaved stands. forest research institute, warsaw: 45-53. występowanie miseczek hymenoscyphus pseudoalbidus w ściole w zamierajacych drzewostanach jesionowych w południowej polsce streszczenie badania prowadzono w sierpniu 2011 roku w 14 drzewostanach jesionowych o różnym wieku na terenie nadl. myślenice oraz w 14 takich drzewostanach w nadl. dynów. w każdym drzewostanie, w jego środkowej części, dokonano oceny nasilenia występowania wybranych symptomów chorobowych u 50 rosnących bezpośrednio obok siebie drzew fraxinus excelsior. ogółem analizie stanu zdrowotnego poddano 1400 drzew. dla oceny częstości występowania miseczek hymenoscyphus pseudoalbidus w każdym drzewostanie zaznaczono 6 (w dwu drzewostanach o małej powierzchni po 5) poletek o wymiarach 0.5 × 0.5 m, w odstępie, co 50 m wzdłuż przekątnej drzewostanu. z poletek zebrano i policzono wszystkie ubiegłoroczne nerwy liściowe i wykształcone na nich miseczki h. pseudoalbidus. ogółem założono 82 poletka w drzewostanach nadl. myślenice i 84 poletka w drzewostanach nadl. dynów. we wszystkich analizowanych drzewostanach jesion wykazywał różnego typu objawy chorobowe: nekrozy na pniach, zamieranie gałęzi, zamieranie wierzchołków oraz całych drzew. na 82 poletkach o wymiarach 0.5 × 0.5 m w nadl. myślenice stwierdzono 1046 nerwów liściowych jesionu. na 72.3% z nich było wykształconych 12477 miseczek h. pseudoalbidus. na 84 poletkach w nadl. dynów stwierdzono 680 nerwów liściowych jesionu. na 98.4% z nich było wykształconych 4166 miseczek tego patogenu. uzyskane dane wskazują na ogromny rezer146 t. kowalski et al. wuar materiału infekcyjnego h. pseudoalbidus wykształcający się w ściole w drzewostanach jesionowych. z szacunkowych przeliczeń wynika, że w analizowanych drzewostanach na powierzchni 1 hektara może dojrzewać w tym samym czasie pomiędzy 370 000 a 13 350 000 miseczek. 2014-02-05t13:34:19+0100 polish botanical society mycodiversity of nature reserves in central italy elena salerni and claudia perini dipartimento di scienze ambientali “g. sarfatti”, università degli studi di siena via p.a. mattioli 4, i-53-100 siena, perini@unisi.it s a l e r n i e., p e r i n i c.: mycodiversity of nature reserves in central italy. acta mycol. 42 (1): 5-19, 2007. seven nature reserves situated in the province of arezzo (tuscany, central italy), presenting various habitats, plants and animals of comunitary interest according to the habitat directive, have been observed from a myco-floristic viewpoint. a synthesis of the results on fungal investigations is given. interesting the finding of rare species such as ramariopsis pulchella and mycena diosma. key words: macromycetes, mycodiversity, nature reserve, habitat directive, tuscany introduction article 2. of the convention on biological diversity (cbd) defines biological diversity as “the variability among living organisms from all sources including, inter alia, terrestrial, marine and other aquatic ecosystems and the ecological complexes of which they are part; this includes diversity within species, between species and of ecosystems”. the slow decline of biological diversity on our planet began approximately 10,000 years ago and has accelerated in the last century: due to human activities, the rate at which species disappear has increased from 100 to 1,000 times the rate of natural extinction. various international organisations have been working on the monitoring and conservation of genetic resources on a planetary scale for many years, benefiting from the support of the fao at various levels. macromycetes, sad to say, have been excluded from all these protection schemes: in italy, for example, there is no specific law for their protection, except for regulations on the picking and selling of edible species. in this context, we report a synthesis of the results of investigations into the mycofloristic composition of seven nature reserves in the province of arezzo (l a g a n à et al. 2003, 2004; p e r i n i et al. 2003; s a l e r n i et al. 2000a). the scientific community’s level of interest in these areas is demonstrated by the numerous acta mycologica vol. 42 (1): 5-19 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 6 e. salerni and c. perini floristic-vegetational and faunistic studies undertaken in recent years, with the aim of improving knowledge of their animal and vegetable heritage and proposing management measures for their conservation, protection and restoration. of particular interest and some of priority interest according to the habitat directive no. 92/43/ cee are unusual habitats such as the humid zones (valle dell’inferno e bandella; ponte a buriano e penna), rocky ophiolitic environments (monti rognosi), mixed tilio-acerion forests (bosco di montalto), the pioneer grassland vegetation of rocky zones (alpe della luna), and at a specific level, both animals (wolf, golden eagle, goshawk, rosalia alpina) and plants (taxus baccata) can be found. the provincial administration strongly encouraged the inclusion of the mycological component in this study, demonstrating a particularly well developed “ecologic conscience” and a sensitivity towards such issues and these organisms in particular that is far-sighted to say the least. material and methods the results reported refer to myco-floristic observation carried out in the seven nature reserves situated in the province of arezzo between 1998 and 2002 (fig. 1). the study was carried out through on-the-spot investigations in autumn (september-november), when the meteorological conditions were deemed optimum for the growth of fruiting bodies (indispensable for recognising the species), and fig. 1. map of the study areas. 1 – valle dell’inferno e bandella; 2 – ponte a buriano e penna; 3 – sasso di simone; 4 – alpe della luna; 5 – montalto; 6 – alta valle del tevere; 7 – monti rognosi. mycodiversity in central italy 7 sporadically in springtime. we recorded all epigeous macromycetes found in the main types of vegetation or particular habitats present in the reserves; samples were often picked for examination under the laboratory microscope in order to check or confirm correct identification. species nomenclature followed that of the dutch check-list (a r n o l d s et al. 1995) taxa not in this list were according to various texts and monographs (b r e i t e n b a c h , k r a n z l i n 1981–2000; m o s e r 1983; r i v a 1988; s t a n g l 1991; a n t o n ì n , n o o r d e l o o s 1993; c o u r t e c u i s s e , d u h e m 1994; b o n 1997; s a r n a r i 1998; c o n s i g l i o , c o n t u 2002). species authorities were abbreviated according to b r u m m i t t , p o w e l l (1992). the species picked were dried and conserved in the herbarium universitatis senensis (siena). for the majority of taxa an abundance value according to j a h n et al. (1967) is given: r (rarus) – species occuring in single exemplares in a amall quantity, n (numerus) – species widespread on the area, single or in several exemplars, a (abundans) – species common on the area in great number of exemplars. area of investigations ponte a buriano e penna and valle dell’inferno e bandella. these two nature reserves are situated the western part of the town arezzo (fig. 1, area 1 and 2) and runs along the middle course of the arno river. according to the t h o r n t h w a i t e classification (1948), the climate is moist subhumid (class c2: mi between 0 and 19.9). from a lithological viewpoint, the area is characterised by fluvio-lacustrine sediments from various periods, fluvial and marsh deposits. the ponte a buriano e penna nature reserve (668 ha) at an altitude between 200 and 260m, runs along the river arno for approximately 7km, beginning slightly upstream of its confluence with the canal maestro della chiana and continuing across the arezzo plain as far as the penna dam. the forest vegetation mainly comprises oak woods of quercus pubescens, as well as other arboreal species such as q. ilex, q. cerris, fraxinus ornus, ostrya carpinifolia and many shrubs (crataegus monogyna, ligustrum vulgare, spartium junceum, etc.) and grasses (cyclamen spp., viola spp., orchis spp., etc.). there are strips of riparian vegetation characterised by poplars, especially black poplars (populus nigra), salix spp., ulmus minor and robinia pseudoacacia along the ditches, gullies and tributaries, and a wide area of marsh vegetation at the confluence of the canal maestro della chiana with the arno, composed of vast stands of common reeds (phragmites australis and smaller areas where rushes (juncus spp.), sedges (carex spp.), yellow irises (iris pseudacorus) can be found (v i c i a n i 1997). the reserve valle dell’inferno e bandella (531 ha) runs along the arno river for approximately 4km, between 180 and 220 m a.s.l.. the deciduos oak woods are mainly composed of quercus cerris woods that also sporadically host other species (q. robur, q. pubescens). hygrophilous riparian vegetation is equally important, comprising salix spp., populus spp., alnus glutinosa, corylus avellana and carpinus betulus growing along the banks of the river arno and its tributaries. a strip of marsh vegetation has formed in the bandella loop in particular, composed of juncus articulatus, j. effusus, j. conglomeratus, nymphaea alba and phragmites australis (r a f f a e l l i 1997). 8 e. salerni and c. perini monti rognosi. the area lies on ophiolites formed by ultramafic igneous rocks (p i c h i s e r m o l l i 1948), a lithology not so common in tuscan areas. the reserve covers a total of 156 ha and is located in the upper basin of the river tevere, north from arezzo (fig.1, area 7). it is characterised by quite steep slopes with visible large rocky outcrops and the altitude varies between 370 and 630m a.s.l.. the climate is mesothermal suboceanic, from humid to subhumid, with a moderate summer deficit (b i g i , r u s t i c i 1984). local factors, such as the dark green colour of the rocks, the lack of soil accumulation and the scarcity of vegetation, may contribute to accentuating the dryness and higher temperatures that characterise the summer months (d e d o m i n i c i s et al. 2001a). the phytogeographic aspect of the area studied is important (d e d o m i n i c i s et al. l.c.) as types of vegetation typical of open areas and well adapted to the relatively inhospitable climate and soil can still be found, despite over seventy years of intense reforestation. the pinewoods planted (pinus nigra, p. pinaster) since the 1930’s on former garigues are generally quite extensive and currently occupy almost 50% of the surface area. the serpentine vegetation is generally confined to steeply sloping zones and rocky outcrops. the garigues have very discontinuous vegetation cover, with large outcrops of bare rock. although limited in area, the steppe grasslands represent very unusual coenoses, with some species of significant conservational interest, such as stipa tirsa, s. etrusca and chrysopogon grillus. semi-mesophilous shrubberies of rosa canina, prunus spinosa, cornus sanguinea, crataegus monogyna, occasional juniperus communis and/or j. oxycedrus and locally abundant spartium junceum are more common. where the pines have not reached, there are mixed woods of turkey oak and european hop hornbeam on the cooler slopes of ophiolite outcrops and downy oak woods in drier and warmer areas. alta valle del tevere (monte nero). this regional nature reserve, at the northern border of tuscany (fig. 1, area 6), covers an area of 470 ha at an altitude of between 740 m and 1240 m a.s.l. and most of the reserve has gradients of over 50%. the climate can be described as mesothermal suboceanic, humid with a slight summer deficit (b i g i , r u s t i c i 1984). from a lithologic point of view, most of the area is located on the marly-arenaceous romagna formation; there is also a strip of macigno, marls, and an undifferentiated complex prevalently composed of clay schists and marly limestones at monte castelsavino. the area has quite a good variety of vegetation. between 700 and 1000 m there are grasslands and shrubberies that originated from the change of use from former agricultural or pastoral land. these limited formations are then replaced by coppices, transitory high forests and conifer plantations. mixed mesophilous montane woods can be found in scattered formation at the same range of altitude, with a prevalence of acer spp. and tilia spp., while the woods in higher zones are clearly dominated by fagus sylvatica l. neutrophilous mountain woods are quite common in this reserve, formed of quercus cerris mixed with ostrya carpinifolia and acer opalus, accompanied by fraxinus ornus, q. pubescens, sorbus aria and occasionally carpinus betulus, a. campestre and prunus avium (d e d o m i n i c i s et al 2001b). sasso di simone. this reserve lies on the apennines in north eastern part of tuscany, bordering with the region emilia romagna and marche (fig.1, area 4). from a lithological point of view, this reserve (1604 ha, 940 – 1221 m a.s.l.) has mycodiversity in central italy 9 three main lithotypes: organogenic calcarenites and marly limestones, chaotic flaky clays and marly-arenaceous flysch. according to the t h o r n t h w a i t e (1948) classification, the climate in this area is humid (class b3, mi between 40 and 60). the forest vegetation is primarily composed of quercus cerris woods associated with carpinus betulus in some places. the forest landscape at the base of the rocky part of the north-facing slope is characterised by mixed woods with a prevalence of fagus sylvatica and the presence of tilia platyphyllos, fraxinus excelsior, acer pseudoplatanus and acer platanoides. the open areas near the “sasso di simone” rock are composed of grassland used for grazing cattle (g o n n e l l i 1997). alpe della luna and bosco di montalto. these two nature reserves are located between alta valle del tevere and sasso simone (fig.1, area 4 and 5), a little bit southern. the climate can generally be described as mesothermal sub-oceanic: humid with a slight summer deficit (b i g i , r u s t i c i 1984). in the two reserves taxus baccata trees can be found. from a vegetational point of view, both areas are of significant phytogeographic interest due to the presence of nuclei of mixed mesophilous montane woods composed of maples, linden and common ash and a few yew trees belonging to the tilio-acerion alliance (d e d o m i n i c i s et al. 2001c,d). alpe della luna covers a total of 1540 ha at an altitude between 600 m at the northern and southern boundaries and 1453 m on the highest ridge. from a geological point of view, the area is characterised by the meeting at the lowest altitude of the macigno del mugello, mainly composed of siliceous sandstones with marls and silty schists, with the marly-arenaceous romagna formation, which is composed of sandstones and marls. the reserve is mainly characterised by coppices and transitory high forests of broad-leaved trees. the woods on the tyrrhenian slopes have a clear prevalence of beech with cardamine enneaphyllos and at a lower altitude the beech is accompanied by other woody species such as crataegus monogyna, laburnum alpinum, quercus cerris and sorbus aria; we should also highlight the presence not only of taxus baccata but also of nuclei of ilex aquifolium. pioneer grasslands grow on both slopes in areas affected by erosion, while the valley floor is home to gorge woods with a prevalence of carpinus betulus, accompanied by beeches, lindens, maples and hazels (d e d o m i n i c i s et al. 2001c). the bosco di montalto reserve occupies a small area (22ha) at an altitude of bereserve occupies a small area (22ha) at an altitude of between 875m and 1061m a.s.l.. the lithology of the area is characterised by the meeting of two geological formations: “alberese”, constituted of calcareous and marlycalcareous sediments, and the “sillano unit”, composed of argillites with lithoid masses of “alberese” and “pietraforte” sandstones (b i n i et al. 1982). most of the reserve’s surface area is covered by thermophilous and meso-hygrophilous beech woods, mainly composed of beeches, turkey oaks and european hop-hornbeam; the presence of tilia platyphyllos, fraxinus exelsior and, although sporadic, of acer pseudplatanus, is also significant. meso-hygrophilous montane woods with turkey oaks and european hornbeam can be found at lower altitudes and on the sillano argillites, with the sporadic presence of taxus baccata (d e d o m i n i c i s et al. 2001d). 10 e. salerni and c. perini results and discussion the seven protected areas observed, with particular emphasis to the interesting habitats, generally show a good mycodiversity (tab. 1) and the presence of fungal ta b l e 1 macrofungal species found in the 7 nature reserves (arezzo, italy) 1 – valle dell’inferno e bandella; 2 – ponte a buriano e penna; 3 – sasso di simone; 4 – alpe della luna; 5 – montalto; 6 – alta valle del tevere; 7 – monti rognosi species 1 2 3 4 5 6 7 agaricus arvensis schaeff. : fr. n agaricus bitorquis (quél.) sacc. r agaricus campester l. : fr. n n agaricus comtulus fr. r agaricus phaeolepidotus (f.h. moeller) f.h. moeller r agaricus porphyrizon p.d.orton r agaricus praeclaresquamosus freeman n agaricus silvicola (vittad.) sacc. n r amanita battarrae (boud.) bon r amanita citrina (schaeff.) pers. f. alba (s. price) quél. n amanita citrina (schaeff.) pers. a a a r amanita mairei foley r amanita pantherina (dc.: fr.) krombh. r n r n amanita phalloides (fr.: fr.) link n n n n amanita rubescens pers.: fr. n r a n amanita vaginata (bull.: fr.) lam. ss. str. * * a n n armillaria mellea s.l. * * * * * armillaria ostoyae (romagn.) herink r armillaria tabescens (scop. : fr.) emel. * astraeus hygrometricus (pers. : pers.) morgan r bisporella citrina (batsch) korf & s.e. carp. n boletus chrysentheron (bull.) ss. str. n n n boletus impolitus fr. * * boletus queletii schulzer * * boletus rubellus krombh. ss. str. * boletus subtomentosus l.: fr. n n n calocybe gambosa (fr. : fr.) singer * calvatia excipuliformis (scop. : pers.) kreisel r r r cantharellus cibarius fr.: fr. * chroogomphus fulmineus (r. heim) courtec. n chroogomphus rutilus (schaeff. : fr.) o.k. mill. a clathrus ruber pers. : pers. r clavaria fragilis holmsk.: fr. r r r clavariadelphus pistillaris (fr.: fr.) donk n * clavariadelphus truncatus (quél.) donk r clavulina coralloides (l. : fr.) j. schröt. a clavulina rugosa (bull. : fr.) j. schröt. r clitocybe alexandri (gillet) gillet * clitocybe candicans (pers. : fr.) p. kumm. * * clitocybe costata kühner & romagn. * * clitocybe fragrans (with. : fr.) p. kumm. * * clitocybe gibba (pers.: fr.) p. kumm. * * a n * clitocybe inornata (sowerby : fr.) gillet * clitocybe metachroides harmaja * clitocybe nebularis (batsch: fr.) p. kumm. n a n a clitocybe obsoleta (batsch : fr.) quél. * mycodiversity in central italy 11 species 1 2 3 4 5 6 7 clitocybe odora (bull. : fr.) p. kumm. n r clitocybe phaeophthalma (pers.) kuyper n n n clitocybe phyllophila (pers. : fr.) p. kumm. * * clitocybe rivulosa (pers. : fr.) p. kumm. r clitocybe subspadicea (j.e. lange) bon & chevassut * clitopilus prunulus (scop. : fr.) p. kumm. a * collybia butyracea (bull.: fr.) p. kumm. * * * a n collybia butyracea (bull.: fr.) p. kumm. var. asema (fr.: fr.) quél. r collybia confluens (pers. : fr.) p. kumm. * collybia cookei (bres.) j. d. arnold * collybia dryophila s.l. * a * n * collybia erythropus (pers.: fr.) p. kumm. * r * collybia fusipes (bull.: fr.) quél. * collybia hariolorum (bull. : fr.) quél. * n collybia peronata (bolton : fr.) p. kumm. n * * coprinus comatus (o. f. müll.: fr.) pers. * n coprinus micaceus (bull. : fr.) fr. n cortinarius aleuriosmus maire * cortinarius allutus fr. * cortinarius anomalus (fr.: fr.) fr. ss. str. * * n cortinarius anserinus (velen.) rob. henry * cortinarius aprinus melot * * n n cortinarius auroturbinatus (secr.) lange r cortinarius bicolor cooke * cortinarius bulliardi (pers.: fr.) fr. * * n * cortinarius calochrous (pers.: fr.) fr. n n n n cortinarius calochrous (pers. : fr.) fr. var. carolii (velen) nezdjm. * cortinarius coerulescens (schaeff.) fr. r * n cortinarius conicus (velen.) rob. henry * cortinarius cotoneus fr. * n * cortinarius delibutus fr. * cortinarius dionysae rob. henry * cortinarius diosmus kühner * * cortinarius duracinus fr. * * * n n n cortinarius glaucopus (schaeff. : fr.) fr. * * cortinarius glaucopus (schaeff. : fr.) fr. var. olivaceus (m.m. moser) quadr. * cortinarius hillieri rob. henry r cortinarius hinnuleus fr. ss.str. * * cortinarius hinnuloides rob. henry * * cortinarius infractus (pers.: fr.) fr. * * * * cortinarius largus fr. * cortinarius lividoochraceus (berk.) berk. n n a cortinarius nanceiensis maire n cortinarius olivaceofuscus kühn. * cortinarius paleaceus fr. ss. str. n n n n cortinarius prasinus (schaeff.) fr. * cortinarius rapaceus fr. * cortinarius rufoolivaceus (pers. : fr.) fr. * n cortinarius safranopes rob. henry n cortinarius salor fr. n cortinarius splendens rob. henry * cortinarius torvus (bull.: fr.) fr. * * * * cortinarius trivialis j. e. lange * * * n a n craterellus cornucopioides (l.: fr.) pers. a a a craterellus lutescens (pers. : fr.) fr. * entoloma ameides (berk. & broome) sacc. r tab. 1 cont. 12 e. salerni and c. perini species 1 2 3 4 5 6 7 entoloma hebes (romagn.) trimbach * entoloma lividoalbum (kühner & romagn.) kubicka * entoloma rhodopolium (fr.: fr.) p. kumm. r r n n r entoloma rhodopolium (fr. : fr.) p. kumm. f. nidorosum (fr.) noordel. n entoloma scabrosum (fr.) noordel. * entoloma sericatum (britzelm.) sacc. * entoloma sericellum (fr. : fr.) p. kumm. * * entoloma serrulatum (fr. : fr.) hesler * entoloma sinuatum (bull. ex pers.: fr.) p. kumm. * * flammulaster carpophilus (fr.) earle r r r ganoderma australe (fr.) pat. r ganoderma lipsiense (batsch) j.f.atk. r ganoderma lucidum (m. a. curtis: fr.) p. karst. r hebeloma crustuliniforme (bull.) quél. ss. str. * n hebeloma mesophaeum (pers.) quél. n * hebeloma sacchariolens quél. ss. str. * * * hebeloma senescens (batsch) berk. & broome * hebeloma sinapizans (fr.) gillet * * * n a n * helvella crispa (scop.) fr. * n * helvella elastica bull. * helvella lacunosa afzel. r * * hemimycena cucullata (pers.: fr.) singer * * * n hemimycena lactea (pers. : fr.) singer * * hohenbuehelia petaloides (bull.: fr.) s. schulz. r humaria hemisphaerica (f.h. wigg.) fuckel r hydnum repandum l.: fr. * * hydnum rufescens fr.: fr. * * * hygrocybe acutoconica (clemençon) singer * n hygrocybe conica (schaeff.: fr.) p. kumm. * * hygrocybe conica (schaeff. : fr.) p. kumm. var. cloroides (malençon) bon * hygrocybe conica (schaeff. : fr.) p. kumm. f. pseudoconica (j. lange) arnolds * hygrocybe conicoides (p.d.orton) orton & watling n hygrocybe pratensis (pers. : fr.) murrill * hygrocybe psittacina (schaeff. : fr.) p. kumm. * hygrocybe russocoriacea (berk. & mill.) p.d. orton & watling r hygrocybe virginea (wulfen: fr.) p. d. orton & watling n * * hygrophoropsis aurantiaca (wulfen : fr.) maire * hygrophorus arbustivus (fr.) fr. * n hygrophorus chrysodon (batsch : fr.) fr. * hygrophorus cossus (sowerby) fr. a * * hygrophorus discoxanthus (fr.) rea * * * a hygrophorus eburneus (bull. : fr.) fr. * a hygrophorus lindtneri m. m. moser * * * hygrophorus penarius fr. * hygrophorus persoonii arnolds n hygrophorus persoonii arnolds var. fuscovinosus (bon) bon * * n hygrophorus roseodiscoideus bon & chevass. * hygrophorus russula (fr.: fr.) quél. * * * hymenoscyphus calyculus (sowerby) w. phillips * inocybe adaequata (britzhelm.) sacc. r inocybe asterospora quél. * * inocybe bongardii (weinm.) quél. * n n * inocybe bongardii (weinm.) quél. var. pisciodora (donadini & riousset) kuyper * inocybe brevicystis métrod ex kuyper * tab. 1 cont. mycodiversity in central italy 13 species 1 2 3 4 5 6 7 inocybe cervicolor (pers.) quél. * * inocybe flocculosa sacc. n inocybe fraudans (britzelm.) sacc. * * * inocybe fuscidula velen. * * inocybe geophylla (fr. : fr.) p. kumm. a a a inocybe geophylla (fr. : fr.) p. kumm. var. lilacina (peck) gillet n inocybe obscurobadia (j. favre) grund & d.e. stuntz n inocybe phaeocomis (pers.) kuyper r inocybe rimosa (bull.: fr.) p. kumm. * * * inocybe sindonia (fr.) p. karst. * inocybe splendens r. heim * inocybe splendens r. heim var. phaeoleuca (kühner) kuyper * inocybe tenebrosa quél. n inocybe terrigena (fr.) kuyper * laccaria amethystina cooke * laccaria laccata s. l. * * * n n laccaria laccata (scop. : fr.) cooke var. pallidifolia (peck) peck * laccaria proxima (boud.) pat. * lactarius acerrimus britzelm. * lactarius azonites (bull.) fr. * lactarius blennius (fr. : fr.) fr. n * * lactarius camphoratus (bull. : fr.) fr. * lactarius chrysorrheus fr. n n n n a n lactarius cimicarius (batsch) gillet n n lactarius decipiens quél. * lactarius deliciosus (l.: fr.) gray * n lactarius ichoratus (batsch) fr. * lactarius insulsus (fr.: fr.) fr. * * lactarius pallidus pers. : fr. * lactarius quietus (fr.:fr.) fr. * * lactarius semisanguifluus r. heim & leclair n lactarius serifluus (d.c.:fr.) fr. * * * * lactarius uvidus (fr. : fr.) fr. * lactarius vellereus (fr. : fr.) fr. * * lactarius zonarius (bull.) fr. * leotia lubrica (scop.) pers. * lepiota aspera (pers.:fr.) quél. r lepiota castanea quél. * lepiota clypeolaria (bull.: fr.) p. kumm. * * * * a n lepiota cristata (alb. & schwein.:fr.) p.kumm. * * * lepista flaccida (j. sowerby: fr.) pat. * * * lepista nuda (fr.: fr.) cooke * * a n lepista sordida (fr. : fr.) singer * leucoagaricus leucothites (vittad.) wasser * leucopaxillus gentianeus (quél.) kotl. * * lycoperdon echinatum pers. : pers. n lycoperdon perlatum pers.: pers. * * * * a n lycoperdon pyriforme schaeff. : pers. * lyophyllum deliberatum (britzelm.) kreise * lyophyllum transforme (britzelm.) singer * * macrolepiota excoriata (schaeff.:fr.) wasser * macrolepiota procera (scop.: fr.) singer * * * marasmius alliaceus (jacq.: fr.) fr. * * marasmius cohaerens (pers. : fr.) cooke & quél. * * marasmius epiphyllus (pers. : fr.) fr. * * marasmius oreades (bolt.:fr.) fr. n * * marasmius torquescens quél. * * tab. 1 cont. 14 e. salerni and c. perini species 1 2 3 4 5 6 7 marasmius wynnei berk. & broome * megacollybia platyphylla (pers. : fr.) kotl. & pouzar * melanoleuca melaleuca (pers. : fr.) murrill * micromphale brassicolens (romagn.) p.d. orton * a mutinus caninus (huds. : pers.) fr. r mycena acicula (schaeff. : fr.) p. kumm. r mycena alba (bres.) kühner * mycena capillaripes peck * mycena diosma krieglst. & schwöbel r n r r mycena epipterygia (scop. : fr.) gray * * mycena galericulata (scop.: fr.) gray * * * mycena galopus (pers.: fr.) p. kumm. n n * * mycena maculata p. karst. * mycena meliigena (berk. & cooke) sacc. * mycena pelianthina (fr.: fr.) quél. * a a n n mycena polygramma (bull.: fr.) gray r r r r mycena pura (pers. : fr.) p. kumm. f. alba (gillet) kühner * * mycena pura (pers.: fr.) p. kumm. n n a a n n mycena rosea (bull.) gramberg * * * * a n n mycena vitilis (fr.) quél. n n n n omphalotus olearius (dc.: fr.) singer * otidea alutacea (pers.) massee r otidea bufonia (pers.) boud. r panaeolus fimicola (fr. : fr.) quél. r panellus stipticus (bull.: fr.) p. karst. * * * paxillus involutus (batsch : fr.) fr. * n phaeomarasmius erinaceus (fr.: fr.) singer * pholiota lucifera (lasch) quél. r pleurotus ostreatus (jacq. : fr.) p. kumm. * pluteus nanus (pers. : fr.) p. kumm. r pluteus thomsonii (berk. & broome) dennis r podophacidium xanthoneum (pers.: fr.) kavina r polyporus varius fr. r r psathyrella candolleana (fr. : fr.) maire n psathyrella tephrophylla (romagn.) bon r psilocybe aeruginosa (m.a. curtis : fr.) noordel. * * * psilocybe fascicularis (huds.: fr.) noordel. n n * * psilocybe sublateritia fr. n * n ramaria flava (schaeff. : fr.) quél. n ramaria stricta (pers.:fr.) quél. n * ramariopsis pulchella (boud.) corner r rhodocybe gemina (fr.) kuyper & noordel. * rhodocybe parilis (fr. : fr.) singer n russula acrifolia romagn. r russula amoenolens romagn. r r russula aurea pers. n russula cessans a. pearson r russula chloroides (krombh.) bres. r n n n r r russula cyanoxantha schaeff.: fr. n r n russula decipiens (singer) svrcek * * russula densifolia gillet * russula fellea (fr. : fr.) fr. r russula foetens pers.: fr. r n n russula fragilis (pers.: fr.) fr. ss. str. n n a a a n russula heterophylla (fr.: fr.) fr. r russula insignis quél. * tab. 1 cont. mycodiversity in central italy 15 species cited in several national and local red lists. some of these are found rarely while other species are more common in some areas. no fungal species listed as threatened at a european level and proposed for inclusion in the bern convention were observed. for instance various larger fungi, red-listed in different euro-various larger fungi, red-listed in different european countries (s c h e m b r i , s u l t a n a 1989; a r n o l d s et al. 1995; b e n d i k s e n et al. 1997; j ä r v a et al. 1998; d i a m a n d i s 2000; g ä r d e n f o r s 2000; wo j e w o d a , ł a w r y n o w i c z 2006), are frequent and abundant in the small “bosco di montalto” nature reserve. these are: collybia hariolorum, cortinarius bulliardii, c. calochrous, c. cotoneus, c. lividoochraceous, c. nanceiensis, hebeloma sinapizans, helvella crispa, hygrophorus arbustivus, h. discoxanthus, h. eburneus, inocybe bongardii, lycoperdon echinatum with numerous carpophores, micromphale brassicolens, mycena diosma, m. pelianthina, rhodocybe parilis, tricholoma atrosquamosum, t. orirubens, t. sulphureum, t. ustale. if adequate information were available, these areas of the provspecies 1 2 3 4 5 6 7 russula luteotacta rea * russula maculata quél. * * russula mairei singer * russula persicina krombh. * * * * * russula risigallina (batsch) sacc. n * n * russula sanguinea (bull.) fr. n russula torulosa bres. a russula vesca fr. * * * russula vinosobrunnea (bres.) romagn. * * * russula xerampelina (schaeff.) fr. * suillus bellini (inzenga) kuntze a suillus collinitus (fr.) kuntze a suillus granulatus (l.: fr.) roussel r a trametes versicolor (l. : fr.) pilát n tricholoma acerbum (bull.: fr.) quél. * * * tricholoma album (schaeff.: fr.) p. kumm. r r n n a tricholoma atrosquamosum (chev.) sacc. n n a r tricholoma basirubens (bon) riva & bon r tricholoma fracticum (britzelm.) kreisel n tricholoma lascivum (fr. : fr.) gillet r tricholoma orirubens quél. n n n tricholoma pessundatum (fr. : fr.) quél. * tricholoma saponaceum (fr.: fr.) p. kumm. n * * tricholoma scalpturatum (fr.) quél. * tricholoma sejunctum (j. sowerby: fr.) quél. r * * n a r tricholoma squarrulosum bres. * * tricholoma sulphureum (bull.: fr.) p. kumm. * * * a * tricholoma terreum (schaeff. : fr.) p. kumm. * * tricholoma ustale (fr.: fr.) p. kumm. n n a n n tricholoma ustaloides romagn. n n tubaria furfuracea (pers.: fr.) gillet * tubaria hiemalis bon n vascellum pratense (pers.:pers.) kreisel * * xerocomus ichnusanus alessio, r.galli & littini r xerocomus roseoalbidus alessio & littini r xerula pudens (pers.) singer * * xerula radicata (relhan: fr.) dörfelt * * * * * xylaria hypoxylon (l.: fr.) grev. * * * tab. 1 cont. 16 e. salerni and c. perini ince of arezzo could become important conservation sites in europe for the more common fungi. many species found belong to a wide ecological range, growing in deciduous or coniferous woods on acidic, basic or neutral substrates, such as amanita phalloides, a. rubescens, a. vaginata ss. str , laccaria laccata s.l. and russula fragilis ss. str. among the simbionts, or collybia butyracea, lepiota clypeolaria, mycena pelianthina, m. pura and m. rosea among the saprotrophs. we should highlight findings of fungi preferring mediterranean areas or at least thermophilous forests, such as boletus impolitus, b. queletii, clathrus ruber, collybia erythropus and hygrophorus russula (a n t o n i n , n o o r d e l o o s 1997; c a n d u s s o 1997; g a l l i 1998) in the montane habitats of the “alpe della luna” nature re-g a l l i 1998) in the montane habitats of the “alpe della luna” nature re1998) in the montane habitats of the “alpe della luna” nature reserve. the presence of two rare boletaceae first observed on the island of sardinia and newly found in the nature reserve of “valle dell’inferno e bandella” is important: xerocomus roseoalbidus alessio and littini and xerocomus ichnusanus alessio, r.galli and littini (g a l l i 1998). x. roseoalbidus, found in 1986 and described as a new taxa the following year, is uncommon and grows on the edge of deciduous woods and in more open areas, in extremely arid climatic conditions. the latter was also first observed in sardinia, then in various locations in the northern apennines, from piedmont to emilia romagna. this thermoxerophilous mushroom becomes more frequent and abundant in hot and dry years (g a l l i 1998). ramariopsis pulchella (boud.) corner, a small coralloid terricolous saprotroph that is reported in various central-northern european studies as being badly threatened because of its great rarity at an international level (a r n o l d s et al. 1995), has also been sighted in the “valle dell’inferno e bandella” reserve. only a few recordings have been cited for tuscany and the most significant of these were in two other protected areas, the forests of berignone-tatti near volterra (province of pisa) and cipresseta di s. agnese, not far from castellina in chianti (province of siena) (r i c c i , p e r i n i 2001). mycena diosma was observed in four out of the seven reserves studied (alpe della luna, montalto, alta valle del tevere, monti rognosi,): this species, characteristic of beech forests (m a a s g e e s t e r a n u s 1992) has never been cited for region of tuscany and is considered rare or very rare by some authors (c o u r t e c u i s s e , d u h e m 1994; b r e i t e n b a c h , k r ä n z l i n 1981–2000). it could be confused by a quick look with the common m. pelianthina, but once collected the characteristic sweetish/tabac smell and withish edge of the lamelle are conclusive evidence that it is m. diosma. clitocybe costata, cortinarius anserinus, c. bulliardii are characteristic species on calcareous substrates according to various authors (b o u d i e r 1901; c a r b i e n e r et al. 1972-73-74; h ø i l a n d 1980; m a l e n ç o n , l l i m o n a , 1980; b r e i t e n b a c h , k r ä n z l i n 1981–2000; b r a n d r u d et al. 1990–1998; g y o s h e v a , va s s i l e v 1994; l a g a n à et al. 1999; s a l e r n i et al. 2000b). their growth in acidophilous quercus cerris woods of the “alta valle del tevere” and not at all on the preferential substrate described can only be explained by the vicinity of the calcareous complex of monte castelsavino. lactarius deliciosus and suillus granulatus, exclusively linked to conifers (b a s s o 1999; g a l l i 1998), have been observed as alien species in the “valle dell’inferno mycodiversity in central italy 17 e bandella” nature reserve, where quercus cerris woods constitute the dominant vegetation. it is well known that conifers, even if they are only sporadic trees, influence the characteristics of the fungal community. the same situation can be reported for “alta valle del tevere”, where planted conifers allow cortinarius allutus to grow (c o n s i g l i o et al. 2004). this is also true for the “monte rognosi” reserve, where important phytogeographic aspects of characteristic garigue and steppe grassland are still present on ultramafic substrates although 50% of the area has been planted with pinus. although the vegetation found is peculiar to serpentine, this is not reflected by the fungal community. in the small open areas where characteristic plants can be found flowering, species linked to conifers such as chroogomphus fulmineus, lactarius deliciosus, russula sanguinea, suillus granulatus have been collected (b a s s o 1999; c o u r t e c u i s s e , d u h e m 1994; g a l l i 1998; s a r n a r i 1998). the situation in the “alpe della luna” reserve is different: in this area a conspic-in this area a conspicuous population of taxus baccata is of importance. this plant is considered a valuable floristic species and has a fragmentary distribution area. very few mycological studies refer to this kind of vegetation (d e v r i e s , k u y p e r 1990). the data found in the area we studied include species linked to the surrounding deciduous woods but none of the mushrooms characteristic of this important tree. on the other hand, abundant open areas and wide grasslands have permitted the identification of fungal species (agaricus arvensis, a. campester, entoloma ameides, hygrocybe pratensis, h. psittacina, h. russocoriacea) that are characteristic of this habitat and considered as a priority vegetation for conservation aspects at a european level (a r n o l d s 2001). also the woods of “sasso di simone” are frequently interspersed with wide open areas, to be maintained, where the fungi agaricus campester and vascellum pratense, characteristic of montane grasslands, grow. conlusions from this short overview of the research carried out in the 7 nature reserves of the province of arezzo, it has emerged how important mycological observations can be for the sustainable management of natural resources: the presence or absence of some fungi could be a good bioindicator of the conservation status of the natural environment (a r n o l d s 1981). a first attempt has been made in order to provide guidelines for appropriate sylvicultural manipulations to safeguard the diversity of larger fungi and to increase the presence of characteristic species. it has been suggested, for example, that reforestation with pines should not take place in areas characterised by serpentine rocks, while more cutting/deforestation should be done in certain zones to increase the steppe garigues. it has also been suggested that some former activities should be reintroduced to encourage the maintenance or expansion of grasslands. the idea of protecting threatened and rare species and the local richness of fungal biota could become a reality in the near future, possibly through the establishment of important plant areas for fungi, as proposed by the european plant conservation strategy (a n d e r s o n 2002; p a l m e r , s m a r t 2001). acknowledgements. the authors give special thanks to the provincial administration of arezzo (councillor for territorial policy – soil defence service), in particular dr. frosini and dr. gusmeroli, for the sensitivity the have shown regarding the mycological heritage present in the reserves, thus supporting the research. 18 e. salerni and c. perini references a n d e r s o n s. 2002. identifying 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(eds) 1989. red data book for the maltese island. interprint ltd., malta. s t a n g l j. 1991. guida alla determinazione dei funghi. 3. inocybe. saturnia s.a.s., trento. t h o r n t h w a i t e c. w. 1948. an approach toward a rational classification of climate. geogr. review 38: 55–94. w o j e w o d a w., ł a w r y n o w i c z m. 2006. red list of the macrofungi in poland. (in:) z. m i r e k , k. z a r z y c k i , w. w o j e w o d a , z. s z e l ą g (eds). red list of plants and fungi in poland 3. ed.: 54-70. w. szafer institute of botany, polish academy of sciences, kraków. v i c i a n i d. 1997. la flora e la vegetazione (in:) riserva naturale di ponte a buriano e penna, sistema delle aree protette della provincia di arezzo. 2014-01-01t11:44:59+0100 polish botanical society a contribution to the morphology and ecology of mycenastrum corium (agaricales) maria ławrynowicz1 and andrzej radwański2 1department of algology and mycology, university of łódź banacha 12/16, pl 90 237 łódź, miklaw@biol.uni.lodz.pl 2institute of geology, university of warsaw żwirki i wigury 93, pl 02 089 warszawa ł a w r y n o w i c z m . , r a d w a ń s k i a .: a contribution to the morphology and ecology of mycenastrum corium (agaricales). acta mycol. 41(1): 73 78, 2006. an interesting collection of mycenastrum corium from suwałki region (ne poland) close to the russian and lithuenian frontiers is presented in this paper. two specimens were found ca. 20 cm under the soil surface. macro and micromorphological features are compared with those of mycenastrum corium growing at the surface. key words: macrofungi, gastromycetoid fungi, ecology of fungi, distribution, hypogeous form introduction mycenastrum corium (guers.) desvaux has been observed in poland since 1889 (b ł o ń s k i 1890). recently, k u j a w a , b u j a k i e w i c z and k a r g (2004) have given a review of 14 localities with a map showing an accumulation of points in the central part of the country. now we report a new and rather unusual site of m. corium discovered in the corner forming the russian, lithuanian and polish borders (fig. 1). basidiocarps were situated below the soil surface (fig. 2). during field work in suwałki region, one of us (a.r.) discovered two basidiocarps determined as mycenastrum corium by the first author. more detailed examination of this collection has showed that fruitbodies growing underground are slightly different in comparison with specimens preserved in herbarium universitatis lodziensis (lod 825) found as occurring on the soil surface at wierzbice on pastures surrounding artificial lake, zalew zegrzyński, 19.06.1972, leg. maria ławrynowicz (c a l o n g e, ł a w r y n o w i c z 1986). the aim of the present paper was to describe and illustrate by scanning electron micrographs morphological characters of recently found specimens of mycenastrum, to show briefly the differences between them and surface growing specimens and to provide an illustrated commentary on the site where the specimens were found. acta mycologica vol. 41 (1): 73-78 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday a contribution to the morphology and ecology of mycenastrum corium 75 agropyron repens, as a whole, evidently affected by human activities; associated fungi: marasmius oreades commonly appearing from time to time; langermannia gigantea and agaricus arvensis occurring locally in wet periods of summer to autumn. distribution and ecology in general mycenastrum corium is known as an epigeous, saprotrophic, nitrophilous, synanthropic ephemeral species (k r e i s e l 1982, 2001; wo j e w o d a 2003). it was found at a variety of sites, from xerothermic to mesophilic and even boggy habitats. wo j e w o d a (2003) has summarised its occurrence in poland as follows: “on dry meadows and pastures, by roads, near houses, xerothermic places, in grass, also on soil with gypsum”. it is indicated as a rare species in poland on the red list (wo j e w o d a , ł a w r y n o w i c z 2006) in the category v (vulnerable). mycenastrum corium is probably much more common, but it grows on sites which are rather seldom visited by mycologists (c a l o n g e , ł a w r y n o w i c z 1986). the species can colonise habitats over a large alltitudial range. in germany up to 330 m a.s.l., in the canaries up to 820 m a.s.l. the total distribution is amphizonal, mainly subcontinental and continental, with isolated localities as far north as the boreal zone, and in mountain regions of tropical east africa (k r e i s e l 1982). some authors, for example: b e n k e r t (2004); d ö r f e l d t and b r e s i n s k y 2003; g r o s s , r u n g e and w i n t e r h o f f (1980); k r e i s e l (1982), write about changing distribution of m. corium in recent years. k r e i s e l (2001, 2006) drew attention to influences of global climate changes on geographical distribution of some macromycetes including mycenastrum corium. k u j a w a et al. (2004) supplies a map with 14 localities known so far in poland. it shows that only one locality: a drained and dried peat bog called kuwasy near grajewo on the biebrza river had been known from north-east poland before the site at wiżajny lake presented in this paper was discovered. ta b l e 1 differences in fruitbodies of mycenastrum m. corium (epigeous) m. corium (hypogeous) basidiocarp white, yellowish to brown when old chocolate brown exoperidium with patches homogenous capillitium long, sharply pointed spines commonly bifurcate with short stumpy spines 76 m. ławrynowicz and a. radwański final remarks 1. mycenastrum has never been mentioned as hypogeous in the literature by mycologists searching for both epigeous gasteromycetes and hypogeous fungi. 2. specimens described differ conspicuously in morphology from known mycenastrum collections. they have a chocolate coloured smooth surface without patches and thicker capillitium hyphae (8-10 μm) which are commonly bifurcate, with short, uniformly distributed stumpy spines. 3. the locality is unique, the most northeastern in poland but this region has been inadequately searched for fungi, especially in ruderal places. 4. taking into account increasing number of localities in the last years, we hope our contribution will stimulate mycologists to pay attention also to hypogeous sites when looking for mycenastrum corium. acknowledgment. the authors thank professor katarzyna turnau and doctor piotr mleczko of the jagiellonian university in kraków for the facilities and assistance in preparing scanning electron micro graphs and doctor david minter (egham) for improving the english version of this paper. this study was supported by the ministry of science and higher education, grant no. 3 po4g 07425. references b e n k e r t d . 2004. die mark brandenburg, auch ein einwanderungsland für pilze. verh. bot. ver. berlin brandenburg 137: 489 514. b ł o ń s k i f. 1890. wyniki poszukiwań florystycznych skrytokwiatowych dokonanych w ciągu lata r. 1889 w obrębie 5 ciu powiatów królestwa polskiego. pamiętn. fizjogr. 10: 129 190. c a l o n g e f. d . , ł a w r y n o w i c z m . 1986. a contribution to the chorology of some gasteromycetes in poland. acta mycol. 18 (2): 161 170. d e s v a u x a . n . 1842. sur le genre mycenastrum du groupe des lycoperdées. ann. sci. nat., 2 sér. 17: 143 147. d ö r f e l t h . , b r e s i n s k y a . 2003. die verbreitung und ökologie ausgewähter makromyceten deutschlands. z. mykol. 2: 177 286. g r o s s g . , r u n g e a . , w i n t e r h o f f w. 1980. bauchpilze (gasteromycetes s. 1.) in der bundesre publik deutschland und westberlin. beih. z. mykol. 2: 79. h a n s e n l . 1962. danish find of mycenastrum corium with notes on its anatomy. bot. tidssk. 58: 204 212. h o m r i c h m . h . , w r i g h t j . e . 1973. south american gasteromycetes. the genera gastropila, lanopila and mycenastrum. mycologia 65: 779 794. k r e i s e l h . 1973. die lycoperdaceae der ddr. bibliotheca mycologica. verl. von j. cramer. lehre. k r e i s e l h . 1982. das vorkommen von mycenastrum corium in ddr. gleditschia 9: 257 269. k r e i s e l h . 2006. global warming and mycoflora in the baltic region. acta mycol. 41(1): 79 94. k u j a w a a . , b u j a k i e w i c z a . , k a r g j. 2004. mycenastrum corium (fungi, agaricales) in poland. polish bot. j. 49 (1): 63 66. p e r r e a u j . , h e i m r . 1971. a propos des mycenastrum représentés ou décrits par n. patouillard. rev. mycol. 36 (2): 81 95. r u d n i c k a j e z i e r s k a w. 1991. flora polska. grzyby (mycota) 23: basidiomycetes, lycoperdales, scle rodermatales, tulostomatales, nidulariales, phallales, podaxales. pwn, warszawa kraków. š e b e k s . 1958. mycenastraceae. (in:) flora čsr: 386 392. praha. w o j e w o d a w. 2003. checklist of polish larger basidiomycetes. (in:) z. m i r e k (ed.) biodiversity of poland 7. w. szafer institute of botany, polish academy of sciences, kraków, pp. 812. w o j e w o d a w. , ł a w r y n o w i c z m. 2006. red list of the macrofungi in poland (in:) z. m i r e k , k. z a r z y c k i , w. w o j e w o d a , z. s z e l ą g (eds). red list of plants and fungi in poland. 3ed.: 53 70. w. szafer institute of botany, polish academy of sciences, kraków. a contribution to the morphology and ecology of mycenastrum corium 77 przyczynek do morfologii i ekologii mycenastrum corium s t r e s z c z e n i e przedmiotem publikacji jest analiza morfologiczna i siedliskowa mycenastrum corium, którego owocniki występowały na głębokości 20 cm pod powierzchnią gleby na wzniesieniu graniczącym z jeziorem wiżajny w pobliżu zbiegu granic polski, litwy i rosji. dwa owocniki odkrył współautor (a. r.) kopiąc ziemię w pobliżu drewnianych zabudowań. cechy taksonomiczne, makro i mikromorfologiczne owocnika rosnącego pod ziemią zo stały opisane oraz przedstawione na rycinach, a także zestawione w tabeli porównawczej z ce chami owocników naziemnych mycenastrum corium. autorzy zwracają uwagę na podziemne występowanie mycenastrum corium na najdalej na północ wysuniętym stanowisku tego grzyba w polsce. 2014-01-01t11:43:36+0100 polish botanical society cercospora berteroae and pseudocercosporella gei, rare anamorphic fungi ewa połeć and małgorzata ruszkiewicz-michalska department of mycology, faculty of biology and environmental protection university of łódź, banacha 12/16, pl-90-237 łódź, ewa_polec@op.pl połeć e., ruszkiewicz-michalska m.: cercospora berteroae and pseudocercosporella gei, rare anamorphic fungi. acta mycol. 47 (1): 21–26, 2012. the paper presents two species of hyphomycetous fungi, presumably mycosphaerella anamorphs, recently collected in central and north-eastern poland. habit and morphology, illustrated with macroand microphotographs, and worldwide distribution of the species are characterized. both species have been collected in poland for the first time. key words: microfungi, mycosphaerella, hyphomycetes, parasites, berteroa, geum, poland introduction the genera cercospora and pseudocercosporella belong to the cercosporella/ramularia complex, which groups 26 genera of colourless or very pale hyphomycetes with holoblastic conidiogenesis (braun 1995). the known teleomorphs of cercospora and pseudocercosporella belong to the ascomycetous genus mycosphaerella s.l. (braun 1995; crous, braun 2003). the genus cercospora was introduced by fresenius in 1863 for the passalora-like species with pluriseptate conidia (crous, braun 2003). many members of the genus are important plant pathogens and often major agents of crop losses worldwide. they occur on a wide range of hosts and can be associated with angiosperms as well as with some gymnosperms and ferns (to-anun, hidayat, meeboon 2011). some species are hyperparasites of rust fungi, e.g., passalora acori (j. m. yen) u. braun & crous, parasitizing the uredospores of uromyces sparganii cooke & peck (shin, kim 2001). according to the recent taxonomic studies of the cercospora species in thailand, the genus is considered to be monophyletic (to-anun et al. 2011). the members of the pseudocercosporella genus are phytopathogens mostly causing leaf spots (shin, kim 2001). the genus was established by deighton over one acta mycologica vol. 47 (1): 21–26 2012 22 e. połeć and m. ruszkiewicz-michalska hundred years after cercospora for the species characterized by unthickened, inconspicuous conidial scars. later braun (1990) narrowed the genus to species with solitary conidia and transferred the taxa with catenate conidia to thedgonia. according to the molecular studies of crous et al. (2012) the genus resides in a large clade together with cercospora, miuraea, phloeospora, septoria and xenocercospora. despite the fact that many members of the cercosporella/ramularia complex are economically important pathogens of major agricultural crops this group of anamorphic fungi is still insufficiently known worldwide (shin, kim 2001; bakhshi, arzanlou, babai-ahari 2012; to-anun et al. 2011). the recently published preliminary list of cercosporoid hyphomycetes known from poland covers 32 species of the genus cercospora and 31 belonging to passalora (świderska-burek 2007). lately, the number of cercosporoid fungi known in the country has been expanded with passalora acericola (liu & guo) u. braun & crous (świderska-burek, mułenko 2010). according to the checklist of polish micromycetes only 6 pseudocercosporella species have been noted in poland so far (świderska-burek 2008). material and methods the original materials with plant organs affected by anamorphic fungi were collected in the urbicoenoses of łódź city and a single specimen originates from bie-biebrza national park. the fresh specimens mounted in lactophenol picric acid solution (fluka) were examined using the microscope nikon eclipse e200; measurements of the morphological structures were made also in tap water. microphotographs of morphological structures of the species were taken with a nikon ds-f1 digital camera. the fungal nomenclature follows crous and braun (2003) and the host plants nomenclature is given after mirek et al. (2002). the vouchers are deposited in the herbarium universitatis lodziensis (lod) in the collection of parasitic fungi labelled as pf. results as the result of mycocoenological research in urbicoenoses of the łódź city two species of anamorphic fungi new for the polish mycobiota were identified, namely cercospora berteroae hollós and pseudocercosporella gei (farl.) u. braun. the first was primarily collected on berteroa incana (l.) dc. in one of the city parks and further on geum urbanum l. in the las łagiewnicki nature reserve. cercospora berteroae and pseudocercosporella gei 23 description of the species cercospora berteroae hollós, ann. mus. nat. hung. 5: 468 (1907) leaf spots amphigenuous, circular or irregular, up to 2.0 mm wide, pale yellow to ochraceous, margin dark (fig. 1a). caespituli amphigenuous, conidiophores in loose to fairly dense fascicles, arising from stromata, simple, straight, erect, smooth, pale yellow to medium brown, darker brown at the base, not branched 65-75 x 5 μm, conidiogenous loci thickened and darkened. conidia solitary, narrowly obclavate, straight, 2-6-septate, hyaline, smooth, truncate at the base, 45-75 x 3.75-5 μm, hila fig. 1. cercospora berteroae (lod 3387 pf): a – host plants with syptoms of fungal infection visible on the leaves; b – conidiophores and conidia (lactophenol picric acid solution; phase contrast). scale bars: a = 1 cm; b = 20 μm. 24 e. połeć and m. ruszkiewicz-michalska thickened, darkened (fig. 1b). morphological features of the specimens generally correspond to those described by chupp (1954, accessed at www.mycobank.org). material examined. on berteroa incana (l.) dc , central poland, łódź, “park im. bolesława chrobre-bolesława chrobrego” park, 12 oct. 2006, leg. d. papierz, lod 3026 pf; ne poland, biebrza national park, obwód ochronny grzędy district, inland dune, sandy grassland, 30 aug. 2012, leg. m. ruszkiewicz-michalska, lod 3387 pf. notes. the species is known only on berteroa incana (brassicaceae) from europe (hungary, lithuania, russia and ukraine) and north america (usa: wisconsin) (crous, braun 2003; farr, rossman 2012). pseudocercosporella gei (farlow) u. braun, nova hedwigia 50: 504 (1990) leaf spots amphigenous, subcircular or angular, up to 6.0 mm in diam., pale yellowish to pale greenish, margin purple, narrow, indistinct (fig. 2a). caespituli epiphyllous, greyish white, punctiform. mycelium internal, forming dense stromata, pale yelowish to pale brownish. conidiophores in dense fascicles, arising from stromata, simple, straight or sometimes curved, subcylindric to slightly geniculate-sinuous, 12.5-25 x 2.5-3 μm, aseptate, hyaline, smooth, conidial scars inconspicuous. conidia solitary, filiform-acicular, 37.5-112.5 x 2.5-3 μm, 1-4-septate, hyaline, smooth, rounded to truncate at the base (fig. 2b). morphological features of the specimen generally correspond to those given by braun (1995). material examined. on geum urbanum l., central poland, łódź, las łagiewnicki reserve, forest roadside, 02 july 2011, leg. a. ozimek, lod 3388 pf. fig. 2. pseudocercosporella gei (lod 3388 pf): a – lower leaves of a host plant with syptoms of fungal infection; b – conidiophores and conidia (lactophenol picric acid solution; phase contrast). scale bars: a = 1 cm; b = 20 μm. cercospora berteroae and pseudocercosporella gei 25 notes. the species has been reported on the members of geum l. (rosaceae) from asia and north america. it was associated with geum aleppicum jacq. (asia, russia), g. radiatum michx. and g. strictum ait. (north america) (braun 1995; farr, rossman 2012). final remarks the presented findings of two new species for poland were the result of studies carried out of in the łódź city, the area greatly subjected to anthropopression. however, cercospora berteroae has been also subsequently confirmed at the protected area in biebrza national park. both fungal species were found on common plant species, which are often parasitized by other micromycetes in poland (mułenko, majewski, ruszkiewicz-michalska 2008). on berteroa incana only biotrophic parasites were recorded, e.g., albugo candida (pers.) kuntze, peronospora berteroae gäum. and erysiphe cruciferarum opiz ex junell. geum urbanum is known to host peronospora gei h. syd., sphaerotheca aphanis (wallr.) u. braun, and two species classically referred to as mycosphaerella anamorphs (e.g., ramularia gei (elliasson) lindr. and septoria gei roberge ex desm.). although cercospora and pseudocercosporella have traditonally been linked to the ascomycetous genus mycosphaerella s.l. (crous, braun 2003), the teleomorphs of the majority of cercosporoid fungi are still unknown and a mycosphaerella state has been proven only for few species (bakhshi et al. 2012). both species reported here have no known teleomorphic connections. there are no mycospharella taxa connected with berteroa species (aptroot 2006) while four species are known to be parasites of geum: sphaerella geicola kalchbr. & cooke, mycospharella koldingensis munk, m. larsenii munk, and m. melanoplaca (dem.) johanson ex oudem. however, none of them was reported in poland so far (mułenko et al. 2008). thus, we assume that species represented by anamorphs reported here are new to polish mycobiota. acknowledgements. the authors are very grateful to professor maria ławrynowicz, curator of the fungal collection of herbarium universitatis lodziensis (lod), for permission to analyse herbarium materials. they include specimens collected by danuta papierz and aleksandra ozimek (msc students supervised by the second author) whose contribution is greatly acknowledged. we thank professor krystyna czyżewska (university of łódź) for providing access to microphotographic equipment as well as two anonymous reviewers for their thorough review of the paper and critical remarks. the studies were partially supported by the ministry of science and higher education (grant no n305 077 32/2708). the first author was also granted in the frame of the project “scholarships to support innovative doctoral research” by the european social fund and the budget as a part of integrated regional operational programme in 2010. 26 e. połeć and m. ruszkiewicz-michalska references aptroot a. 2006. mycosphaerella and its anamorphs: 2. conspectus of mycosphaerella. cbs biodiversity series 5: 1–231. bakhshi m., arzanlou m., babai-ahari a. 2012. comprehensive check list of cercosporoid fungi from iran. plant pathology & quarantine 2 (1): 44–55. braun u. 1990. studies on ramularia and allied genera (iii). nova hedwigia 50: 499–521. braun u. 1995. a monograph of cercosporella, ramularia and allied genera (phytopathogenic hyphomycetes). 1. ihw – verlag, eching. chupp c. 1954. a monograph of the fungus genus cercospora. ithaca, new york. published by the author. crous p. w., braun u. 2003. mycosphaerella and its anamorphs: 1. names published in cercospora and passalora. cbs biodiversity series 1: 1–571. crous p. w., braun u., hunter g. c., wingfield m. j., verkley g. j. m., shin h.-d., nakashima c., groenewald j. z. 2012. phylogenetic lineages in pseudocercospora. stud. mycol. 75: 35-114. farr d. f., rossman a.y. 2012. fungal databases, systematic mycology & microbiology laboratory, ars, usda [retrieved 13.09.2012, from the database at: http://nt.ars-grin.gov/fungaldatabases/]. mirek z., piękoś-mirkowa h., zając a., zając m. (eds). 2002. flowering plants and pteridophytes of poland. a checklist. (in:) z. mirek (ed.). biodiversity of poland 1. w. szafer institute of botany, polish academy of sciences, kraków, 442 pp. mułenko w., majewski t., ruszkiewicz-michalska m. (eds). 2008. a preliminary checklist of micromycetes in poland. (in:) z. mirek (ed.). biodiversity of poland 9. w. szafer institute of botany, polish academy of sciences, kraków, 752 pp. shin h. d., kim j. d. 2001. cercospora and allied genera from korea. nat. inst. agric. sci. tech., suwon, korea, 302 pp. świderska-burek u. 2007. preliminary list of cercosporoid fungi from poland. mycotaxon 102: 5–8. świderska-burek u. 2008. genera pseudocercospora & pseudocercosporella. (in:) w. mułenko, t. majewski, m. ruszkiewicz-michalska (eds). a preliminary checklist of micromycetes in poland (in:) z. mirek. (ed.). biodiversity of poland 9. w. szafer institute of botany, polish academy of sciences, kraków: 461–462. świderska-burek u., mułenko w. 2010. passalora acericola – a rare cercosporoid species found for the first time in poland. mycotaxon 113: 351–354. to-anun c., hidayat i., meeboon j. 2011. genus cercospora in thailand: taxonomy and phylogeny (with a dichotomous key to species). plant pathology & quarantine 1 (1): 11–87. cercospora berteroae i pseudocercosporella gei, rzadkie gatunki grzybów anamorficznych streszczenie podczas badań nad pasożytniczymi mikromycetes prowadzonych w granicach aglomeracji łódzkiej stwierdzono występowanie dwóch gatunków grzybów anamorficznych, które nie były wcześniej notowane na obszarze naszego kraju. są to cercospora berteroae hollós pasożytujący na berteroa incana (l.) dc. i pseudocercosporella gei (farl.) u. braun na geum urbanum l. materiał został zebrany w parku im. bolesława chrobrego w łodzi, w rezerwacie las łagiewnicki położonym również w granicach miasta oraz w biebrzańskim parku narodowym. w pracy zawarto opisy cech morfologicznych grzybów i prezentujące je mikrofotografie. zestawiono także dane o ich rozmieszczeniu na świecie. 2014-01-02t11:59:30+0100 polish botanical society diversity of fungi colonizing and damaging leaves of pontic azalea azalea pontica maria kowalik department of plant protection, university of agriculture in kraków 29 listopada 54, pl-31-425 kraków, m.kowalik@ogr.ur.krakow.pl kowalik m.: diversity of fungi colonizing and damaging leaves of pontic azalea azalea pontica. acta mycol. 48 (2): 227–236, 2013. the research aimed at verification of fungi species colonizing phyllosphere of pontic azalea azalea pontica l. and at comparison of the fungi species composition: – in the natural stand in the kołacznia nature reserve, – in arboretum collections at bolestraszyce and rogów. 600 fragments of healthy, infected and fallen leaves of pontic azalea were collected for mycological analyses. the species forming the largest number of colonies identified from the healthy leaves were: a. alternata, ph. cyclaminis, e. nigrum, ph. medicaginis and b. cinerea, from infected leaves: a. alternata, e. nigrum, ph. cyclaminis, s. fimicola, t. viride and a. phaeospermum, whereas: e. nigrum, a. alternata, s. fimicola, ph. cyclaminis and b. cinerea were isolated from the fallen leaves, which indicates that a majority of fungi persistently colonize the leaves during vegetation period and damage them, which leads to defoliation. colonization of pontic azalea phyllosphere in arboreta by more numerous fungi colonies and species than under conditions of natural sites evidences their increased pressure in the arboreta environment. key words: pontic azalea, arboretum, reserve, health status, leaves introduction pontic azalea azalea pontica l., i.e. yellow rhododendron [rhododendron luteum sweet, syn. r. flavum g. don (anioł-kwiatkowska 2003)] is a critically endangered species included in the polish red book of plants (kaźmierczakowa, zarzycki 2001). its only natural stand in poland is located in the kołacznia nature reserve at wola zarzycka near leżajsk. the seeds collected on the natural site were used to cultivate the specimens which are growing in the collection of arboretum and department of physiography in bolestraszyce and in the arboretum of the warsaw university of life sciences (sggw) in rogów (piórecki, dubiel 2009; piórecki, zarzycki 2010). acta mycologica vol. 48 (2): 227–236 2013 doi: 10.5586/am.2013.024 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 228 m. kowalik health status of azalea shrubs is affected by cultivation conditions resulting from the specific site requirements and disease agents. identification of the organisms colonizing phyllosphere of pontic azalea, including pathogens causing health disturbances, is a comparative study comprising plants under conditions of natural sites and in arboreta. identification of the population and species diversity of fungi in pontic azalea phyllosphere makes possible following the results of colonization and damaging of leaves. the aim of the research was verification of fungi species colonizing the phyllosphere of azalea pontica and comparison of fungi species composition on the natural site and in the arboreta. materials and methods observations of the of pontic azalea health status were conducted in the third decade of may, july, august and september 2011 on a site in the kołacznia nature reserve, on the collection of the arboretum in bolestraszyce (se poland) and on the collection of arboretum in rogów (central poland). on each date 10 healthy leaves (green, without obvious disease symptoms), infected leaves (showing necrotic symptoms) and fallen leaves were collected from the three sites. a total of 1800 leaf fragments were collected for mycological analyses. the leaf fragments were disinfected in 70% ethanol. isolation and cultivation of mycobiota were conducted according to standard methods applied in mycology (kowalik 2008). for taxonomic identification of the mycobiota the following keys were used: guba (1961); domsch et al. (1980); sutton (1980); ellis, ellis (1987) and rifai (1987). the basis of classification was the system of kirk et al. (2008) and the authors’ epithets by fungal species names were verified according to index fungorum (2012). on the basis of fungi specification considering the share of individual species in the total fungi community, they were classified to the group of dominants (constituting >5% of the entire community), influents (1-5%) and accessory fungi (<1%). similarity coefficient (sőrensen index) was calculated for the analysed sites, comprising the number of fungi species (kowalik 1993). results altogether 2120 fungi colonies belonging to 64 species were isolated from the plant material: healthy, infected and fallen leaves of azalea pontica collected on the natural site in the kołacznia nature reserve, on the collection of the arboretum in bolestraszyce and on the collection of arboretum in rogów. there were 568 colonies comprising 39 species isolated from healthy green leaves (tab. 1), 657 colonies and 40 species from infected leaves (with necrotic symptoms) (tab. 2) and 895 colonies and 52 species from fallen leaves (tab. 3). the largest number of fungi colonies and species existed on healthy azalea leaves in the kołacznia nature reserve, whereas the least number in the arboretum in bolestraszyce. these leaves were in the first place colonized by: alternaria alternata, diversity of fungi colonizing and damaging leaves of pontic azalea 229 phialophora cyclaminis, epicoccum nigrum, phoma medicaginis and botrytis cinerea. these fungi were classified to the dominant group. cladosporium sphaerospermum, trichoderma viride, pestalotiopsis sydowiana, isaria fumorosea, mortierella alpina, m. parvispora, khuskia oryzae, trichoderma koningii, sordaria fimicola, mammaria echinobotryoides, penicillim expansum, p. waksmanii, truncatella truncata and other were classified to the influent group, whereas the other 10 species were identified as accessory fungi (tab. 1). table 1 fungi colonizing healthy leaves of pontic azalea azalea pontica fungus k oł ac zn ia re se rv e b ol es tr as zy ce ar bo re tu m r og ów ar bo re tu m to ta l pe rc en ta ge [% ] alternaria alternata (fr.) keissl. 8 28 23 59 10.39 apiospora montagnei sacc. 12 12 2.11 arthrinium euphorbiae m.b. ellis 9 9 1.58 arthrinium phaeospermum (corda) m.b. ellis 3 3 0.53 aspergillus versicolor (vuill.) tirab. 1 1 0.18 aureobasidium pullulans (de bary) g. arnaud 1 7 8 1.41 botrytis cinerea pers 31 1 32 5.63 chaetomim globosum kunze 2 2 0.35 cladosporium cladosporioides (fresen.) g.a. de vries 1 1 9 11 1.94 cladosporium herbarum (pers.) link 5 4 9 1.58 cladosporium sphaerospermum penz. 8 5 11 24 4.23 coleophoma rhododendri syd. 1 1 0.18 davidiella macrocarpa crous, k. schub. & u. braun 9 3 12 2.11 epicoccum nigrum link 6 27 11 44 7.75 giberella pulicaris (fr.) sacc. 2 2 0.35 ilionectria radicicola (gerlach & l. nilson) p. chaverri & salgado 1 1 0.18 isaria fumorosea wize 3 7 12 22 3.87 khuskia oryzae h.j. hadson 7 12 19 3.34 mammaria echinobotryoides ces. 14 14 2.46 mortierella alpina peyronel 14 6 20 3.52 mortierella parvispora linnem. 14 7 21 3.70 mucor hiemalis f. hiemalis wehmer 1 1 2 0.35 parahoma chrysanthemicola (hollós) gruyter, aveskamp & verkley 1 1 2 0.35 penicillium expansum link 13 13 2.29 penicillium waksmanii k.m. zalessky 5 8 13 2.29 pestalotiopsis sydowiana (bres.) b. sutton 20 3 23 4.05 phialophora cyclaminis j.f.h. beyma 35 13 1 49 8.63 phoma eupyrena sacc. 2 4 6 1.06 phoma exigua (desm.) gruyter, aveskamp & verkley 1 1 2 0.35 phoma medicaginis malbr. & roum. 11 26 37 6.51 sordaria fimicola (roberge ex desm.) ces. & de not 11 11 1.94 thanatephorus cucumeris (a.b.frank) donk 3 3 0.53 trichoderma koningii oudem. 17 17 2.99 trichoderma pseudokoningii rifai 11 11 1.94 trichoderma viride pers. 9 4 14 27 4.75 truncatella truncata (lev.) steyaert 4 8 12 2.11 umbelopsis isabellina (oudem.) w. gams 9 9 1.58 umbelopsis ramanniana (möller) w. gams 2 2 0.35 umbelopsis vinacea (dixon-stew.) arx 2 1 3 0.53 total 206 174 188 568 100.00 230 m. kowalik table 2 fungi colonizing infected leaves of pontic azalea azalea pontica fungus k oł ac zn ia re se rv e b ol es tr as zy ce ar bo re tu m r og ów ar bo re tu m to ta l pe rc en ta ge [% ] alternaria alternata (fr.) keissl. 21 85 56 162 24.65 arthrinium euphorbiae m.b. ellis 2 2 0.30 arthrinium phaeospermum (corda) m.b. ellis 16 11 7 34 5.17 arthrinium sphaerospermum fuckel 2 2 0.30 aureobasidium pullulans (de bary) g. arnaud 1 3 4 0.60 botrytis cinerea pers. 11 7 18 2.74 chaetomim globosum kunze 2 12 14 2.13 cladosporium cladosporioides (fresen) g.a. de vries 2 8 3 13 1.98 cladosporium herbarum (pers.) link 4 6 10 1.52 cladosporium sphaerospermum penz. 1 15 8 24 3.65 davidiella macrocarpa crous, k. schub. & u. baun 2 2 0.30 epicoccum nigrum link 14 32 47 93 14.16 fusarium culmorum (w.g. sm.) sacc. 2 1 3 0.46 fusarium oxysporum schltdl. 2 3 5 0.77 fusarium poae (peck) wollenw. 4 4 0.60 giberella pulicaris (fr.) sacc. 7 7 1.07 ilionectria radicicola (gerlach & l. nilson) p. chaverri & salgado 4 4 0.60 khuskia oryzae h.j. hads. 2 2 0.30 mammaria echinobotryoides ces. 6 1 7 1.07 mortierella alpina peyronel 6 7 13 1.98 mortierella parvispora linnem. 7 3 10 1.52 mucor hiemalis f. hiemalis wehmer 1 1 0.15 paraconiothyrium minitans (w.a. campb.) verkley 4 4 0.60 paraphoma chrysanthemicola (hollós) gruyter, aveskamp & verkley 3 3 0.46 paraphoma fimeti (brunaud) gruyter, aveskamp & verkley 5 5 0.77 penicillium expansum link 1 1 0.15 penicillium waksmanii k.m. zalessky 9 7 16 2.44 pestalotiopsis sydowiana (bres) b. sutton 17 17 2.59 phialophora cyclaminis j.f.h. beyma 16 18 16 50 7.61 phoma exigua (desm.) aveskamp, gruyter & verkley 1 4 5 0.77 phoma medicaginis malbr. & roum. 7 7 1.07 phoma putaminum speg. 4 4 0.60 pleurostomophora richardsiae (nannf.) l. mostert, w. gams & crous 1 4 5 0.77 rhizopus stolonifer (ehrenb.) vuill. 6 6 0.92 sordaria fimicola (roberge ex desm.) ces & de not 23 8 11 42 6.40 talaromyces wortmannii c.r. benj. 2 2 0.30 thanatephorus cucumeris (a.b. frank) donk 7 7 1.07 trichoderma viride pers. 29 4 2 35 5.33 umbelopsis isabellina (oudem.) w. gams 1 8 1 10 1.52 umbelopsis ramanniana (möller) w. gams 4 4 0.60 total 184 237 236 657 100.00 diversity of fungi colonizing and damaging leaves of pontic azalea 231 table 3 fungi colonizing fallen leaves of pontic azalea azalea pontica fungus k oł ac zn ia re se rv e b ol es tr as zy ce ar bo re tu m r og ów ar bo re tu m to ta l pe rc en ta ge [% ] actinomucor elegans (eidam) c.r. benj. et hesselt 3 3 0.34 alternaria alternata (fr.) keissl. 21 47 51 119 13.29 apiospora montagnei sacc. 2 2 0.22 arthrinium euphorbiae m.b. ellis 1 1 0.11 arthrinium phaeospermum (corda) m.b. ellis 1 1 0.11 arthrinium sphaerospermum fuckel 1 1 0.11 aspergillus ustus (bainier) thom et church 2 2 0.22 botrytis cinerea pers. 32 28 60 6.70 cadophora malorum (kidd et beaumont) w. gams 5 5 0.56 calonectria morganii crous, alfenas & m.j. wingf. 1 1 2 0.22 chaetomim globosum kunze 6 6 0.67 cladosporium cladosporioides (fresen.) g.a. de vries 2 4 3 9 1.01 cladosporium sphaerospermum penz. 7 7 0.78 epicoccum nigrum link 72 72 67 211 23.57 fusarium culmorum (w.g. sm.) sacc. 2 3 5 0.56 fusarium flocciferum corda 2 2 0.22 fusarium oxysporum schltdl. 4 4 0.45 giberella pulicaris (fr.) sacc. 2 2 0.22 giberella tricincta el-gohll, mcritchie, schoult. et ridings 5 5 0.56 humicola fuscoatra var. fuscoatra traaen 4 4 0.45 humicola grisea traaen 7 3 10 1.12 ilionectria radicicola (gerlach & l. nilson) p. chaverri & salgado 2 5 7 0.78 isaria fumosorosea wize 2 2 0.22 mammaria echinobotryoides ces. 5 5 0.56 mortierella alpina peyronel 2 16 18 2.01 mucor hiemalis f. hiemalis wehmer 5 7 12 1.34 oidiodendron griseum robak 1 1 0.11 oidiodendron tenuissimum (peck) s. huges 1 4 5 0.56 paraphoma chrysanthemicola (hollós) gruyter, aveskamp, gruyter & verkley 1 2 3 0.34 penicillium citrinum thom 1 2 3 0.34 penicillium expansum link 3 3 0.34 pestalotiopsis sydowiana (bres.) b. sutton 1 7 8 0.89 phialophora cyclaminis j.f.m. beyma 42 14 11 67 7.49 phoma destructiva plowr. 1 1 0.11 phoma eupyrena sacc. 9 9 1.01 phoma exigua (desm.) aveskamp, gruyter & verkley 4 28 32 3.58 phoma herbarum westend. 11 11 1.23 phoma medicaginis malbr. & roum. 18 16 34 3.80 phoma putaminum speg. 1 3 4 0.45 pleospora azalea (voglino) priest 13 13 1.45 pleurostomophora richardsiae (nannf.) l. mostert, w. gams & crous 1 1 0.11 rhizopus stolonifer (ehrenb.) vuill. 16 11 27 3.02 scopulariopsis brumptii salv.-duval 6 6 0.67 scopulariopsis chartarum (g. sm.) f.j. morton et g.sm. 11 11 1.23 sordaria fimicola (roberge ex desm.) ces. et de not 30 28 22 80 8.94 talaromyces wortmannii c.r. benj. 3 3 0.34 trichoderma koningii oudem. 6 6 0.67 trichoderma pseudokoningii rifai 2 2 0.22 trichoderma viride pers. 9 5 14 1.56 truncatella truncata (lév.) steyaert 3 15 18 2.01 umbelopsis isabellina (oudem.) w. gams 12 5 7 24 2.68 umbelopsis ramanniana (möller) w. gams 4 4 0.45 total 244 315 336 895 100.00 232 m. kowalik infected leaves, showing symptoms of necrosis collected in arboreta were colonized by a comparable number of fungi colonies, while leaves in bolestraszyce and on the natural site in kołacznia by a similar number of fungi species (tab. 2). dominant colonies: a. alternata, e. nigrum, ph. cyclaminis, s. fimicola, t. viride and arthrinium sphaerospermum were isolated from these leaves in a prevailing number. 14 species which were classified to the influents included: c. sphaerospermum, b. cinerea, chaetomium globosum, p. sydowiana, m. alpina and p. waksmanii. the least numerous fungi colonies and species existed on the fallen leaves gathered in the kołacznia reserve, whereas the largest number was found in the rogów arboretum (tab. 3). epicoccum nigrum, a. alternata, s. fimicola, ph. cyclaminis i b. cinerea colonies dominated in fungi community isolated from the fallen azalea leaves. in the influent group the most numerous were: ph. medicaginis, ph. exigua and rhizopus stolonifer. a total of 634 colonies comprising 37 species were identified on the leaves in three analyzed sites in the kołacznia reserve, 726 colonies and 43 species in the bolestraszyce arboretum and 28 species within 760 colonies in the arboretum in rogów. similarity indices between individual sites of pontic azalea sites, computed for the communities of fungi species (tab. 4), evidence the greatest number of common species (50-56%) occurring on infected and healthy leaves in the arboreta in bolestraszyce and rogów. the same high similarity index (over 48%) was computed for infected leaves collected in the kołacznia reserve and in both arboreta. fungi communities isolated from fallen leaves differed considerably by their species composition, the least number of common species (28%) occurred in the fungi community isolated from leaves in the kołacznia reserve and rogów arboretum. discussion azalea shrubs growing on natural sites, in collections of arboreta or botanical gardens, but also in city green areas or in household gardens are under pressure of various pathogenic organisms, most frequently fungi and fungus-like organisms (kita, mazurek 2003; kowalik 2008; kowalik et al. 2012). the importance of phyllosphere, in which a competition exists between pathogens and saprotrophs resulting in diseases leading to premature plant defoliation, is emphasized in discussion on plant health status. table 4 similarity coefficient for fungi communities isolated from leaves of pontic azalea azalea pontica on three sites leaves similarity coefficient [%] kołacznia bolestraszyce kołacznia rogów bolestraszyce rogów healthy 39.13 45.83 50.00 infected 48.88 48.97 56.52 fallen 44.44 28.57 31.03 total 47.79 40.79 44.59 diversity of fungi colonizing and damaging leaves of pontic azalea 233 generally, fungi species colonizing healthy pontic azalea leaves were saprotrophs from the genera of apiospora, arthrinium, isaria, khuskia, mammaria, mortierella, trichoderma and umbellopsis. the share of necrotrophs, such as a. alternata, e. nigrum and s. fimicola in the total fungi community isolated from azalea leaves on the natural site in the kołacznia reserve was small, whereas s. fimicola did not colonize azalea leaves in the arboreta at all. while comparing fungi communities isolated from pontic azalea leaves and evergreen rhododendron leaves (kowalik 2009), it was noticed that also numerous saprotrophs from the acremonium, aspergillus, chaetomium and humicola genera colonized azalea leaf blades. asymptomatic colonization of pontic azalea by b. cinerea in the kołacznia reserve in may during the vegetation period resulted in the occurrence of grey mould on the bushes with visible symptoms of necrosis on the edges of leaf blades. the number of fungi in the community colonizing healthy laves of azalea growing on the natural site in the kołacznia reserve was much higher than in the arboreta in bolestraszyce or rogów, however, the number of pathogen colonies in this community was low. a similar relationship was described by kita and mazurek (2003) who compared azalea health status in the arboretum in wojsławice and botanical garden in wrocław (sw poland), however ascribing the effect of polluted air on reduction of pathogenic fungi seems unfounded. diversified similarity indices for fungi communities isolated from healthy leaves indicate a species diversity of fungi colonizing azalea leaves on natural sites and in arboreta. alternaria alternata and e. nigrum dominated on infected, necrotic azalea leaves collected in the arboreta in bolestraszyce and rogów, which confirms previous research results (kowalik 2009; kowalik et al. 2011). the same fungi, but also coelophoma empetri, humicola fuscoatra, h. grisea, p. expansum, penicillium verrucosum, p. sydowiana, septoria azalea, s. fimicola, umbelopsis isabellina, as well as fungi from cylindrocarpon, fusarium, mortierella, phialophora, phoma, trichoderma and other genera were found on necrotic evergreen rhododendron and azalea leaves (kowalik, muras 2007; kowalik 2008; kowalik et al. 2010a; 2010b; 2012). the results obtained confirm the pathogenicity and aggressiveness of many of these fungi species, previously registered on rhododendron leaves. leaf damage due to thanatephorus cucumeris, a pathogen only sporadically isolated from rhododendron leaves (kowalik et al. 2010b) is visible in the presented investigations. high similarity indices for fungi communities isolated from infected leaves indicate that necrosis of azalea leaves on three analyzed sites was caused by a group of pathogenic species, including necrotrophs. numerous investigations (kita, mazurek 2003; kowalik, muras 2007; kowalik et al. 2010 a; 2011) documented the presence and role of necrotrophs in the process of causing necrotic blots, dieback and intensified, premature falling of leaves. it was demonstrated that toxic fungus a. alternata played the dominant role. presented investigations draw attention to a low number of this species on healthy and infected leaves on the natural site in kołacznia in comparison with the arboreta in bolestraszyce and rogów. this situation may be due to vicinity of other species from ericaceae family – a reservoir of pathogenic fungi (kowalik, sagan 2005; kowalik, wandzel 2005). isolation of almost twice larger number of fungi colonies and species from prematurely fallen leaves, in comparison with healthy leaves, evidences an intensified 234 m. kowalik colonization and damaging of leaf blade by various fungi species. species, which earlier caused leaf necroses, among others e. nigrum, a. alternata, s. fimicola, ph. cyclaminis i b. cinerea played the main role. these fungi constituted almost 60% of the identified isolates. equally big share of a. alternata, e. nigrum, s. fimicola and p. expansum was registered in previous papers by kowalik et al. (2011, 2012). presence of fungi from actinomucor, mucor, rhizopus, trichoderma, mortierella and umbelopsis genera on fallen leaves may be connected with the plant age, represented by senile specimens with rotten, moist shoots. presence of over a dozen fungi species of between once and thrice frequency is noticeable in the fungi communities isolated from fallen leaves, which may evidence an accidental colonizing of fallen leaves by the species living in soil. isolation of pathogenic fusarium, giberella, ilionectria, phoma fungi from infected and fallen pontic azalea leaves was corroborated in the literature of the subject (kita, mazurek 2003; kowalik, muras 2007; kowalik 2008; kowalik et al. 2010a; 2010b; 2011; 2012). a comparison of p. sydowiana (syn. pestalotia sydowiana) and truncatella truncata (syn. pestalotia truncata) in causing necrotic symptoms on the leaves of azalea and evergreen rhododendron leaves (kowalik 2008, 2009; kowalik et al. 2010a, 2010b, 2012) revealed that these fungi were more often colonizing healthy leaves of pontic azalea, than infected or fallen leaves. the highest percentage differences between similarity indices for fallen azalea leaves evidence a considerable diversity of mycobiota colonizing this kind of leaves. conclusions 1. fungi communities existing in pontic azalea phyllosphere on the natural site in the kołacznia nature reserve and on the collection of the arboretum and department of physiography in bolestraszyce, as well as on the warsaw university of life sciences (sggw) arboretum collection in rogów differed with the species composition and number of colonies. 2. the species whose colonies were isolated in largest numbers from healthy leaves of pontic azalea comprised: a. alternata, ph. cyclaminis, e. nigrum, ph. medicaginis and b. cinerea, from infected leaves: a. alternata, e. nigrum, ph. cyclaminis, s. fimicola, t. viride and a. phaeospermum, and from fallen leaves: e. nigrum, a. alternata, s. fimicola, ph. cyclaminis and b. cinerea, indicating that majority of them persistently colonized and damage leaves during vegetation period leading to their premature falling. 3. colonization of pontic azalea phyllosphere in the arboreta at bolestraszyce and rogów by a much higher number of fungi species and colonies than in conditions of the natural site in kołacznia evidences their intensified pressure in the arboreta environment. diversity of fungi colonizing and damaging leaves of pontic azalea 235 references anioł-kwiatkowska j. 2003. wielojęzyczny słownik florystyczny. wydawnictwo uniwersytetu wrocławskiego. domsch k.h., gams w., anderson t.h. 1980. compendium of soil fungi. acad. press. london, new york, toronto, sydney, san francisco. ellis m.b., ellis j.p. 1987. microfungi on land plants. croom helm. london, sydney. guba e.f. 1961. monograph of monochaetia and pestalotia. harvard univ. press. cambridge, mass. index fungorum 2012. (www.index fungorum.org), access 15.12.2012. kaźmierczakowa r., zarzycki k. 2001. polska czerwona księga roślin. paprotniki i rośliny kwiatowe. instytut botaniki im. w. szafera pan, kraków: 281-283. kirk p.m., cannon p.f., minter d.w., stalpers j.a. 2008. ainsworth & bisby’s dictionary of the fungi.10th edition. cabi, wallingford, uk. kita w., mazurek j., 2003. skład gatunkowy fyllosfery różanecznika w ogrodzie botanicznym we wrocławiu i w arboretum w wojsławicach. erica polonica 14: 25-36. kowalik m. 1993. grzyby gleby inicjalnej industroziemnej rekultywowanego w kierunku rolnym i leśnym zwałowiska kopalni siarki “machów”. zesz. nauk. ar w krakowie, rozpr. hab. 180: 1-78. kowalik m. 2008. fungi and fungi-like oomycetes isolated from affected leaves of rhododendron. acta mycol. 43 (1): 21-27. kowalik m. 2009. bioróżnorodność grzybów występujących w fyllosferze różanecznika zimozielonego rhododendron l. zesz. probl. post. nauk roln. 539: 341-348. kowalik m., kierpiec b., bonio j., żołna m. 2011. fungi inhabiting spots and necroses on the leaves of azaleas (rhododendron) in the botanical garden of the jagiellonian university. phytopathologia 62: 41-48. kowalik m., kierpiec b., żołna m. 2012. fungi living at the fallen leaves of rhododendron and azalea (rhododendron l.). acta sci. pol., hortorum cultus 11 (2): 161-166. kowalik m., muras p. 2007. grzyby zasiedlające opadłe liście różanecznika. rocz. ar w poznaniu 383. ogrodnictwo 41: 69-73. kowalik m., muras p., kierpiec b., żołna m. 2010a. zdrowotność liści różaneczników zawsze zielonych rhododendron l. zesz. probl. post. nauk roln. 551: 117-123. kowalik m., muras p., żołna m., kierpiec b. 2010b. grzyby wyosobnione z nekrotycznych plam na liściach różaneczników zawsze zielonych rhododendron l. zesz. probl. post. nauk roln. 554: 49-55. kowalik m., sagan a. 2005. fungi causing dying out heather in permanent plantings. acta mycol. 40 (2): 191-195. kowalik m., wandzel a. 2005. grzyby powodujące zamieranie sadzonek wrzosu. acta agrobotanica 58 (2): 237-242. piórecki j., dubiel e. 2009. volynian polesia – main source of the yellow azalea (rhododendron luteum sweet) in european garden and parks. rocznik polskiego towarzystwa dendrologicznego 57: 29-32. piórecki j., zarzycki k. 2010. arboretum bolestraszyce. przewodnik. bolestraszyce. rifai m. a. 1987. a revision of genus trichoderma. mycol. papers 116: 1-56. sutton b. c. 1980. the coelomycetes. cmi, kew, surrey. 236 m. kowalik różnorodność grzybów zasiedlających i uszkadzających liście azalii pontyjskiej azalea pontica streszczenie obserwacje stanu zdrowotnego azalii pontyjskiej azalea pontica przeprowadzono w 2011 roku na stanowisku naturalnym w rezerwacie kołacznia, w kolekcji arboretum i zakładu fizjografii w bolestraszycach oraz w kolekcji arboretum sggw w rogowie. celem badań była weryfikacja gatunków grzybów zasiedlających fyllosferę azalii pontyjskiej oraz porównanie składu gatunkowego grzybów na naturalnym stanowisku i w arboretach. analizie mykologicznej poddano liście zdrowe, porażone i opadłe. stwierdzono, że zbiorowiska grzybów bytujących w fyllosferze azalii pontyjskiej na stanowisku naturalnym i w arboretach różniły się składem gatunkowym i liczbą kolonii. gatunkami wyodrębnionymi w największej liczbie kolonii ze zdrowych liści azalii pontyjskiej były: a. alternata, ph. cyclaminis, e. nigrum, ph. medicaginis i b. cinera, z liści porażonych: a. alternata, e. nigrum, ph. cyclaminis, s. fimicola, t. viride i a. phaeospermum, a z liści opadłych: e. nigrum, a. alternata, s. fimicola, ph. cyclaminis i b. cinerea, co wskazuje, że większość z nich w okresie wegetacji stale zasiedla liście i uszkadza je, co prowadzi do ich przedwczesnego opadania. zasiedlenie porażonych liści azalii w arboretach przez porównywalną liczbę kolonii i gatunków grzybów (w tym patogenów), znacznie większą niż w rezerwacie, może świadczyć o wpływie sąsiadujących roślin żywicielskich na ich stan zdrowotny. kolonizacja fyllosfery azalii pontyjskiej w arboretach w bolestraszycach i rogowie przez dużo większą liczbę kolonii i gatunków grzybów, niż w warunkach naturalnego stanowiska w kołacznii, świadczy o wzmożonej presji grzybów w środowiskach arboretów. 2013-12-20t14:47:50+0100 polish botanical society rare and interesting species of psathyrella found in the tatra national park anna ronikier department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, a.ronikier@ib-pan.krakow.pl r o n i k i e r a .: rare and interesting species of psathyrella found in the tatra national park. acta mycol. 42 (1): 85-92, 2007. three species of the genus psathyrella are described and illustrated in the paper. one of them: p. hydrophiloides is new to poland, while two others: p. conopilus and p. murcida are new to the polish carpathians. key words: agaricales, psathyrella conopilus, psathyrella hydrophiloides, psathyrella murcida, the tatra mts, poland introduction the genus psathyrella (fr.) quél. is represented in poland by 52 species, among which only three, namely: p. candolleana (fr.: fr.) maire, p. piluliformis (bull.: fr.) p.d.orton and p. prona (fr.) gillet, are common in poland. remaining species are known from few localities in the country, some of them are considered as threatened (wo j e w o d a 2003). relatively low number of taxa reported from poland to date and their rarity result probably from the difficulties in identification of species, their variability and long-lasting lack of sufficient monographs and keys. only in late 70. and 80. of the last century the excellent work of k i t s v a n wa v e r e n (1982, 1985) brought a number of papers ordering and clarifying the taxonomical complexity within the genus psathyrella. a great number of species newly described by him, re-description of others and many additional comments on morphological variability and ecological preferences, provided in his papers, improved the knowledge of the genus and facilitated identification of some less known taxa. for the last fifteen years several new to poland species of the genus psathyrella were reported (e.g. wo j e w o d a , h e i n r i c h , k o m o r o w s k a 1999). acta mycologica vol. 42 (1): 85-92 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 86 a. ronikier materials and methods in a framework of the study devoted to the diversity of agarics and boletes of the sarnia skała massif in the tatra national park (r o n i k i e r 2005) a few interesting species of psathyrella were found. the fungi were collected during the years 2000–2003 in a beech forest of lower montane belt (dentario glandulosae-fagetum) of a calcareous massif of sarnia skała. the descriptions of macroand microcharacters provided below are based exclusively on the collected material. references to colours in descriptions of psathyrella conopilus and p. murcida follow k o r n e r u p and wa n s c h e r (1965). the spores were measured in 5% koh. the spores for measurements were taken from a spore-print, from natural spore deposit present at stipe apex or only mature (the darkest) spores lying on a gill surface were measured. drawings of microcharacters were made with the aid of drawing tube (nikon y-idt). all measurements were done directly through the microscope (under oil immersion objective), not from the drawings. the distribution map provided for p. conopilus is based on literature data only and includes the new locality reported in this paper. because usually there is no description of specimens which localities are mentioned in the source literature, the information could not be verified – no herbarium material was revised. the material is deposited at the herbarium of the w. szafer institute of botany, polish academy of sciences, kraków (kram) or the herbarium of the t. chałubiński museum, zakopane (zamu). description of species psathyrella conopilus (fr.: fr.) a.pear son et dennis, 1948. trans. brit. mycol. soc. 31: 185. syn. p. subatrata (batsch) gillet ss. m.m.moser. (figs 1, 3). description: cap 20–30 mm in diameter, 15–25 mm high, broadly conical, hygrophanous, brown (5e4, 5e5, 5f4, 5f5), translucently striate, pallescent on drying to beige (4b3, 4b4), without red tints when dry, surface smooth. veil absent. lamellae distant, 3 mm broad, narrowly adnate, very dark brown to black, edge white ciliate. stipe 70–130 × 3 mm, clavate, base swollen up to 0.5 cm, cream (4a2), silky smooth, finely pruinose at apex. context very thin, cream, smell and taste none. spores 15–17 × 8–9 µm ellipsoid, smooth, thick-walled, with distinct central germ pore 2–2.5(3) µm in diameter, grey-olive-brown in 5% koh, warm brown in 10% nh4oh. basidia 26–30 × 11–12 µm, broadly clavate, with 4 sterigmata and basal clamp. cheilocystidia 35–60 × 11–20 µm, lageniform, utriform, ventricose, hyaline, thin-walled, numerous. pleurocystidia none. caulocystidia present at stipe apex, 37–72 × 10–20 µm, cylindrical to lageniform with rounded and often slightly broadened apex. pileipellis a hymeniderm, built up of clavate cells 27–45 × 20–26 µm; red-brown, thick-walled setae 100–300 µm long present in pileipellis. hymenophoral trama pale yellow-brown in 10% nh4oh. clamps present. specimens examined: s poland, the western tatra mts, the sarnia skała massif, mouth of the dolina strążyska valley, orographically right side of the valley, n slopes of the grześkówki ridge, alt. 950 m a.s.l., dentario glandulosae-fagetum, on soil, n 49°16’43’’ e 19°56’27’’, 14.10.2003, leg. a. ronikier, kram f-54148. psathyrella in the tatra national park 87 notes: psathyrella conopilus is a very remarkable fungus easy to distinguish from other species. it is characterized macroscopically by brown conical cap without red tinges, long stipe, slightly swollen at base and absence of veil. the most distinctive microscopical character is the presence of thick-walled setae in pileipellis. the species belongs to the section subatratae (romagn.) ex singer and it is the only european member of this section. for the synonymy of the species see k i t s v a n wa v e r e n (1985) and e n d e r l e (1992). fig. 1. psathyrella conopilus (fr.: fr.) a. pear son et dennis, coll. kram f-54148; a – caulocystidia, b – spores, c – cheilocystidia, d – basidia, e – pileipellis; scale bar = 10 µm. 88 a. ronikier distribution in poland: the species was reported from several localities in poland (fig. 2). the interpretation of the species in older literature is not clear. in most source data only the name of species is given, without any description of specimens. sometimes the description is provided, but it is too short or it disagrees with the modern interpretation of the species (e.g. spores are smaller). without revision of herbarium specimens it is not possible to verify such data. for this reasons older records (before 1945) are marked with empty circle in the map. the locality from the tatra mts is the first one in the polish carpathians. psathyrella hydrophiloides kits van wav., 1982. persoonia 11 (4): 488. (figs 4a, 5). description: cap 20–25 mm in diameter, 15 mm high, broadly conical, hygrophanous, chocolate-brown, not translucently striate, pallescent on drying to ochraceousbeige without red tints, surface densely radially wrinkled. veil not seen. lamellae distant, 2 mm broad, very narrowly adnate, dark brown, edge finely white ciliate. stipe 15 × 4 mm, whitish, very finely fibrilose, silky, with dense woolly mycelium forming volva-like structure at base, hollow. context thin, wood colour, smell and taste none. spore-print dark brown. fig. 2. distribution of psathyrella conopilus in poland; white circle – localities reported before 1945 (some of them may be doubtful), black circle – after 1945, black triangle – new locality. psathyrella in the tatra national park 89 spores 5.5–6 × 3–3.5 µm ellipsoid in face view, slightly phaseoliform in profile view, smooth, thick-walled, with a distinct germ pore 1–1.5 µm in diameter, greyishbrown in koh, pale brown in 10% nh4oh. basidia difficult to observe, insufficient to make measurements and drawings. cheilocystidia 18–25 × 5–10 µm, utriform, some of them swollen at apex, hyaline, thin-walled, numerous. pleuro cystidia similar, 30–35 × 8.5–16 µm. hymenophoral trama yellowish-brown in 10% nh4oh. clamps numerous in cortical layer of the stipe. specimens examined: s poland, the western tatra mts, the sarnia skała massif, lower part of the dolina spadowiec valley, alt. 950 m a.s.l., dentario glandulosaefagetum, on wood, under side of a fallen trunk of fagus sylvatica covered by rhizomorphs of armillaria sp., n 49°16’36’’ e 19°57’7’’, 01.12.2000, leg. a. ronikier, kram f-54446. notes: p. hydrophiloides belongs to the section hydrophilae romagn. ex singer emend. kits van wav., for which chief characters are, among others, small, relatively pale spores and pigmented hymenophoral trama (k i t s v a n wa v e r e n 1982). the species is very close to p. piluliformis (bull.: fr.) p.d.orton (=p. hydrophila (bull.) maire apud maire et werner) from which it differs by a scanty veil, solitary occurrence and a distinct germ pore in spores. in the original description k i t s v a n wa v e r e n (1982) mentions the terrestrial habitat of the species. on the other hand, he states that specimens of drosophila appendiculata var. piluliformis ss. ricken described by k ü h n e r and r o m a g n e s i (1953) belongs to the same species. although k ü h n e r and r o m a g n e s i (1953) did not mention the habitat of the variety, the fungus depicted in the fig. 494 is presented on a piece of wood (black element at the stipe base). the specimens from the tatra mts. were also collected on wood. distribution in poland: the species is new to poland. psathyrella murcida (fr.) kits van wav., 1985, persoonia suppl. 2: 281. (figs 4b, 6). description: cap 20–40 mm in diameter, first hemispherical, then broadly conical, paraboloid, hygrophanous, grey-brown, chocolate-brown (5e6, 5f6, 5d5, 5e5), translucently striate, pallescent on drying to ochraceous, ochraceous-beige without red tints (4c4, 4c5), surface smooth. veil present, white, fugacious, visible only in young specimens. lamellae moderately distant, very broad (6 mm), broadly adnate, chocolate-brown (5d4, 5e4), edge white ciliate or almost even. stipe 40–100 × 3–6 fig. 5. psathyrella hydrophiloides kits van wav., coll. kram f-54446; a – spores, b – cheilocystidia, c – pleurocystidia; scale bar = 10 µm. 90 a. ronikier mm, cylindrical, sometimes slightly swollen at base, hollow, whitish-cream, dirty white, silky, apex finely pruinose. context thin, beige in cap, whitish-beige in stipe, smell and taste none. spore-print very dark brown, almost black. spores 9–11(12) × 5–6 µm ellipsoid, smooth, thick-walled, with small but distinct central germ pore about 1.5 µm in diameter, grey-olive-brown in koh, sordid brown in 10% nh4oh. basidia 18–21 × 6.5–10 µm, broadly clavate, with 4 sterigmata. cheilocystidia 28–46 × 9–15 µm, obtusely fusiform, lageniform, cylindrical, hyaline, thin-walled, numerous. pleurocystidia 43–72 × 7–16 µm, narrowly obtusely fusiform, hyaline, abundant. caulocystidia present at stipe apex, 43–62 × 9–30 µm, cylindrical, lageniform, narrowly fusiform or sphaeropedunculate. hymenophoral trama pale beige. clamps present. veil composed of hyphae. specimens examined: s poland, the western tatra mts, the sarnia skała massif: upper part of the dolina spadowiec valley, orographically right side of the valley, alt. 1090 m a.s.l., dentario glandulosae-fagetum, on litter (leaves of fagus sylvatica), fig. 6. psathyrella murcida (fr.) kits van wav., coll. kram f-50386; a – pleuro cystidia, b – spores, c – cheilo cys tidia, d – caulocystidia, e – basidia; scale bar = 10 µm. psathyrella in the tatra national park 91 n 49°16’24’’ e 19°57’7’’, 8.11.2000, leg. a. ronikier, kram f-50386; lower part of the dolina strążyska valley, orographically right side of the valley, alt. 960 m a.s.l., dentario glandulosae-fagetum, on litter (leaves of fagus sylvatica), n 49°16’36’’ e 19°56’17’’, 24.11.2000, leg. a. ronikier, kram f-50458; lower part of the dokram f-50458; lower part of the do-lower part of the dolina białego valley, at the droga pod reglami hiking trail, alt. 940 m a.s.l., dentario glandulosae-fagetum, on litter (leaves of fagus sylvatica), n 49°16’44’’ e 19°57’23’’, 14.10.2003, leg. a. ronikier, kram f-54443; the same, 3.10.2002, leg. a. ronikier, zamu 4407; the dolina spadowiec valley, western slope (right side of the valley), alt. 1020 m a.s.l., dentario glandulosae-fagetum, on litter (leaves of fagus sylvatica), n 49°16’26’’ e 19°57’01’’, 3.10.2002, leg. a. ronikier, kram f-54444; lower part of the dolina strążyska valley, the grześkówki ridge, alt. 960 m a.s.l., dentario glandulosae-fagetum, on litter (on small piece of wood of ?fagus sylvatica), n 49°16’39’’ e 19°56’26’’, 3.10.2002, leg. a. ronikier, kram f-54445. notes: ö s t r a d i u s (1992) pointed out that psathyrella murcida in a modern sense is not in agreement with fries’ original description of agaricus murcidus referring to a small, fragile species placed in vicinity of a. gracilis fr. this author, however did not propose any nomenclatural solutions. the disagreement with a protologue had been already noticed by k ü h n e r and r o m a g n e s i (1953) who followed ricken’s interpretation of the species. p. murcida is considered by modern authors (e.g. k i t s v a n wa v e r e n 1985; ve s t e r h o l t , k n u d s e n 1992; h o r a k 2005) as a relatively large sized species with fugacious veil, obtusely-fusiform cystidia and large spores, occurring in fagus forests. p. murcida is the most similar to p. fusca (schumach.) a. pearson from which it differs, among others, in the shape and size of pleurocystida. it occurs also later in season than the latter species. p. murcida is a late-autumn fungus, fairly common in fagus forests of the investigated area where it was collected since beginning of october till end of november. distribution in poland: psathyrella murcida was reported from a few localities in poland. some records, however, seem to be doubtful. as it is believed to be a beech wood species (k i t s v a n wa v e r e n 1985; k ü h n e r , r o m a g n e s i 1953; ve s t e r h o l t , k n u d s e n 1992) occurring late in the season, records from forests without admixture of fagus sylvatica: galio-carpinetum – date of collection not mentioned and querco-carpinetum – collected in 22 aug. (b u j a k i e w i c z , f i k l e w i c z 1963; l i s i e w s k a , p o ł c z y ń s k a 1998) probably refer to other species of deciduous forests. apart from the new locality given in this paper, the fungus was reported from beech forest and pine forest with admixture of fagus sylvatica and abies alba in roztocze region, eastern poland (d o m a ń s k i 1997, 1999). acknowledgments: my thanks are due to professor barbara gumińska (kraków) for permission to use her database and to mr. marian wysocki and mr. jacek wieser (kraków) for kindly providing me with a background of a map of poland elaborated by them. this work is a result of studies carried out within a grant no. 6 p04g 083 20 financed by the polish state committee of scientific research (kbn). 92 a. ronikier references b u j a k i e w i c z a . , f i k l e w i c z g. 1963. grzyby wyższe lasów dębowo-grabowych okolic opalenicy (pow. nowy tomyśl, wielkopolska). bad. fizjogr. pol. zach., b 12: 277–300. d o m a ń s k i z . 1997. przyczynek do znajomości flory mikologicznej roztocza (msc). d o m a ń s k i z . 1999. nowe stanowiska rzadkich i interesujących grzybów w polsce (msc). e n d e r l e m . 1992. studien in der gattung psathyrella ii. beiträge zur kenntnis der pilze mitteleuropas 8: 85–102. h o r a k e . 2005. röhrlinge und blätterpilze in europa. kryptogamenflora, band iib/2, 6. aufläge. spektrum akad. verlag, münchen. k i t s v a n wa v e r e n e. 1982. notes on the genus psathyrella-viii. persoonia 11 (4): 473–508. k i t s v a n wa v e r e n e. 1985. the dutch, french and british species of psathyrella. persoonia 2 suppl. k o r n e r u p a . , wa n s c h e r j . h . 1965. farver i farver [methuen handbook of colour]. politikens forlag, københavn. k ü h n e r r . , r o m a g n e s i h . 1953. flore analytique des champignons supérieurs. masson et cie éditeurs, paris. l i s i e w s k a m., p o ł c z y ń s k a m . 1998. changes in macromycetes of the oak-hornbeam forests in the “dębina” reserve (northern wielkopolska). acta mycol. 33 (2): 191–230. ö s t r a d i u s l. 1992. on the interpretation of psathyrella murcida and p. fusca. persoonia 14 (4): 543– 546. r o n i k i e r a . 2005. bioróżnorodność grzybów agarykoidalnych i boletoidalnych sarniej skały w tatrzańskim parku narodowym. w. szafer institute of botany, polish academy of sciences, kraków (phd thesis). ve s t e r h o l t j., k n u d s e n h. 1992. psathyrella (fr.) quél. (in:) l . h a n s e n , h . k n u d s e n (eds). nordic macromycetes 2. nordsvamp, copenhagen: 236–252. w o j e w o d a w. 2003. checklist of polish larger basidiomycetes. (in:) z . m i r e k (ed.). biodiversity of poland 7. institute of botany, polish academy of sciences, kraków. w o j e w o d a w., h e i n r i c h z., k o m o r o w s k a h . 1999. macromycetes of oak-lime-hornbeam woods in the niepołomice forest near kraków (s poland) – monitoring studies. acta mycol. 34 (2): 201–266. rzadkie i interesujące gatunki z rodzaju psathyrella znalezione w tatrzańskim parku narodowym s t r e s z c z e n i e w niniejszej pracy przedstawiono trzy interesujące gatunki z rodzaju psathyrella. jeden z nich, p. hydrophiloides, nie był dotychczas podawany z terenu polski. jest to rzadki grzyb również w europie. dwa kolejne gatunki zostały podane po raz pierwszy dla polskich karpat. p. murcida jest rzadkim grzybem związanym z bukiem, natomiast p. conopilus występuje głównie w lasach liściastych. ten ostatni gatunek znany jest w polsce z około 30 stanowisk. zamieszczono dokładne opisy cech makroi mikroskopowych przedstawianych gatunków, jak również zdjęcia ich owocników oraz rysunki cech mikromorfologicznych. dla p. conopilus opracowano mapę rozmieszczenia w polsce na podstawie dotychczas opublikowanych danych. fig. 3. psathyrella conopilus (fr.: fr.) a. pear son et dennis, coll. kram f-54148; scale bar = 10 mm. fig. 4a – psathyrella hydrophiloides kits van wav., coll. kram f-54446; b – psathyrella murcida (fr.) kits van wav., coll. kram f-50386; scale bar = 10 mm. 2014-01-01t11:45:24+0100 polish botanical society fungi occurrence on seeds of field pea joanna marcinkowska department of plant pathology, warsaw university of life sciences nowoursynowska 159, pl-02-776 warszawa, joanna_marcinkowska@sggw.pl marcinkowska j.: fungi occurrence on seeds of field pea. acta mycol. 43 (2): 77–89, 2008. seeds of four edible cultivars of pisum sativum and three fodder harvested in 2004-2006 from eight localities, scattered in all region suitable for pea production in poland, were evaluated for fungi occurrence on cn agar medium in petri plates. the highest number (27) of species was isolated in 2004, while the lowest (16) in 2006. number of fungi inhabiting seeds was influenced mainly by environmental conditions of locality and years. alternaria alternata dominated in each sample of 450 seeds. species of penicillium contaminated seeds as the next and infection by stemphylium botryosum was at similar level. fusarium poae was the most often occurring species of this genera. pea specific pathogens: mycosphaerella pinodes, phoma pinodella and ascochyta pisi infected more seeds in 2004 and 2005 than 2006, and at the last season only a. pisi was noted. in general, level of infection by those pathogens was low, reaching on an average only 2.56%, with the highest for a. pisi, and the lowest for m. pinodes. key words: different fungi, occurrence, intensity, ascochyta blight, pea cultivars introduction studies concern fungi transmission by pisum sativum l. seeds were conducted in different countries, like australia (bretag et al. 1995), canada (xue et al. 1997; morrall et al. 2005), france (roger et al. 1999; fougereux et al. 2006), poland (grzelak, iłłakowicz 1973; filipowicz 1976; marcinkowska 1997), but mainly for incidence of ascochyta blight fungi. some reports (skolko et al. 1954; czyżewska 1976; filipo­ wicz 1976; marcinkowska 1997; 1998) were also performed for saprobic fungi, since they could had been able to cause seed destruction and thus decreased plant stands of peas (filipowicz 1976), soybean (marcinkowska, schollenberger 1979), or parsley (nowicki 1997). saprobic fungi are also important because some of them may produce secondary metabolites harmful for people and animals (kozakiewicz 1990; 1992). fungi inhabiting pea seeds in poland were already described, but on genotypes issued earlier. now the increasing importance of dry pea seed production for edible acta mycologica vol. 43 (1): 77–89 2008 78 j. marcinkowska and animal feeding (fodder) purposes caused breeders interest in releasing new cultivars of field pea. seeds of recently introduced cultivars were the objective for evaluation of any fungi occurring on them. materials and methods seeds of four edible and three fodder cultivars, six bred in poland and one in czech, registered between 1995-2006, were tested for fungi occurrence (tab. 1). seeds for evaluation were harvested in 2004, 2005 and 2006 from plants cultivated in fields of 14 localities situated in seven different regions of poland, suitable for pea production. cultivars of each type were planted in eight localities, six different and two the same, since at cicibór (c) and kawęczyn (ka) were tested edible cultivars requiring better soils for cultivation and also fodder one of light soil requirements. a sample of 450 seeds of each cultivar, collected at 8 localities were studied every year. surface sterilized seeds (marcinkowska, boros, in print) were placed on coon’s (cn) agar medium into a petri plate (pp) 10 cm in diameter. a plate contained 15 seeds. evaluation was done in two equal series (15 pp x 15 seeds equals 225), first started in late december, second by the end of february. the same incubation conditions were provided for seeds of both series (marcinkowska 1997). identification of fungi was done following different keys (marcinkowska 1998; 2003). list of fungi occurred on seeds was given in table 2. data were taken on the eight day since plating. number of identified species was counted together for both series and changed for percent of a sample. for statistical evaluation percentage data were transformed according to bliss (tp) and used in statgraphics plus programme. majority of occurring fungi (tabs 3, 4), that means nine common species and 4 genera of more than one species (aspergillus, cladosporium, fusarium, penicillium), were covered by statistical analyses. statistical analysis were also done for comparison of frequency of three species responsible for ascochyta blight and alternaria alternata on tested seeds ta b l e 1 names of cultivars and origin of tested seeds no cultivar r.y. locality of planting cultivars part of poland edible fodder edible krzyżewo (k) north-east 1. ramrod 1995 marianowo (mr) north-east 2. set 2000 cicibór (c) cicibór (c) central-east 3. tarchalska 2004 kawęczyn (ka) kawęczyn (ka) central 4. terno 2006 cz kościelna w. (kw) central masłowice (m) central fodder sulejów (s) central 1. hubal 2005 lubinicko (l) central-west 2. sokolik 2001 chrząstowo (ch) central-north 3. zagłoba 2000 radostowo (r) central-north bobrowniki (b) north-west rarwino (rr) north-west wyczechy (w) north-west tomaszów b. (tb) south-west abbreviations: r. y. – year of cultivar registration; cz – cultivar bred in czech republic fungi occurrence on seeds 79 (tabs 5, 6). incidence of fungi on seeds was evaluated according to fisher’s least significant difference (lsd) procedure, the method used to discriminate among the means at the 95% confidence level. results on evaluated seeds of 7 cultivars, 4 edible and 3 fodder, 27 species of fungi were identified in 2004, 22 in 2005 and 16 in 2006 (tab. 2). statistical analysis of data concern majority of species occurring on seeds indicated that number of fungi inhabiting edible cultivars and fodder ones depended significantly on their species and year (tab. 3). significance of differences between locality and cultivar was various for both groups of cultivars. the most often isolated fungus from seeds, over cultivars, localities and years, was a. alternata, as often as penicillium spp. from fodder cultivars, but for edible one the next was s. botryosum and penicillium spp., both of homogenous group, of lower intensity, to which partly belonged also a. pisi (tab. 4). the other species responsible ta b l e 2 fungi found on seeds of tested cultivars during 2004-2006 no species years 2004 2005 2006 1 alternaria alternata (fr.) keissler + + + 2 ascochyta pisi libert + + + 3 aspergillus flavus link + + – 4 aspergillus niger tieg. + + + 5 botrytis cinerea pers.: fr. + + + 6 chaetomium globosum kunze: fr + + – 7 cladosporium cladosporioides fres. + + + 8 cladosporium herbarum (pers.: fr.) link + + – 9 epicoccum purpurascens link – + – 10 fusarium avenaceum (corda: fr.) sacc. + + – 11 fusarium equiseti (corda) sacc. + – – 12 fusarium oxysporum schlecht. + + + 13 fusarium poae (peck) woll. + + + 14 fusarium solani (mart.) sacc. + + – 15 fusarium sp. + + + 16 mucor hiemalis wehm. + – + 17 mycosphaerella pinodes (berk. et blox.) vesterg. + + – 18 papulaspora sp. + + – 19 penicillium claviformae bainier + – + 20 penicillium expansum link: fr. + + + 21 phoma pinodella (l.k.jones ) morgan-jones et burch + + – 22 rhizoctonia solani kühn + + + 23 rhizopus stolonifer (ehrenb.: fr.) vuill. var. stolonifer + + + 24 sclerotinia sclerotiorum (lib.) de bary + + + 25 stemphylium botryosum wallr. + + + 26 trichocladium asperum harz + – – 27 trichothecium roseum link + – – 28 ulocladium atrum preuss + + + 29 non-sporulating fungi + + + abbreviations: (+) a fungus occurred; (-) not occurred 80 j. marcinkowska for ascochyta blight, p. pinodella and m. pinodes, occurred less frequently, being on similar level for both cultivar types, with lower percentage on edible for the first fungus. fusarium spp. represented by f. avenaceum, f. equiseti and f. oxysporum inhabited less seeds compared to f. poae but more than f. solani. infection by the last fungus was the lowest for fodder cultivars, while by b. cinerea for edible one. when statistical analysis was done only for a. alternata, the species most often inhabiting seeds, and the specific pea pathogens, m. pinodes (syn. didymella pinodes (berk. et blox.) petrak) anamorph ascochyta pinodes l. k. jones), a. pisi and p. pinodella, significant differences were found again for fungi occurring on seeds of both cultivar groups, and for edible – on localities, but for fodder – in years (tab. 5). a. alternata dominated, reaching respectively 10.29 and 8.33 percent, on edible and fodder cultivar seeds compared to species responsible for ascochyta blight, which were covered by second homogenous group (tab. 6). localities influenced also seed contamination but statistically proved differences were noted only for edible cultivars both when majority of fungi and only ta b l e 3 analysis of variance for seed contamination of edible and fodder cultivars by majority of fungi species occurring over cultivars, localities and years source of variation four edible three fodder cultivars d.f. p­value (level of significans) h. ns d.f. p­value (level of significans) h. ns fungus 12 0.0000 significant 5 12 0.0000 significant 3 locality 7 0.0000 significant 3 7 0.2249 non­significant 1 cultivar 3 0.7071 non­significant 1 2 0.0003 significant 2 year 2 0.0557 significant 2 2 0.0000 significant 3 abbreviations: d. f. – degrees of freedom; h. ns– number of homogenous groups ta b l e 4 multiple range test for different fungi species occurrence on seeds of both types of cultivars (a mean over cultivars, localities and years) fungi species cultivars four edible three fodder percent (tp) h. gr (5) percent (tp) h. gr (3) botrytis cinerea 1.34 a 3.94 a b rhizoctonia solani 1.84 a b 3.18 a fusarium solani 2.15 a b 2.00 a mycosphaerella pinodes 2.84 a b 2.82 a aspergillus spp. 2.88 a b 3.31 a fusarium spp. 3.12 a b 3.43 a phoma pinodella 3.19 a b 4.38 a b cladosporium spp. 4.06 b c 3.48 a fusarium poae 4.27 b c 3.97 a b ascochyta pisi 5.81 c d 4.22 a b penicillium spp. 6.79 d 7.81 c stemphylium botryosum 7.42 d 5.02 b alternaria alternata 10.92 e 8.49 c abbreviation: h.gr – homogenous groups fungi occurrence on seeds 81 a. alternata and ascochyta blight species were considered, being the highest at radostowo, krzyżewo and rarwino (tab. 7). at sulejów and chrząstowo no infection by fungi responsible for ascochyta blight was found. frequency of majority of fungi incidence on seeds of edible cultivars was statistically documented between years 2004 and 2005 compare to lower one, of hot and dry season 2006 (tab. 8). significant differences were also found for fodder cultivars both contaminated by all fungi with the highest in 2005 and the lowest in 2006, and also a. alternata and ascochyta blight species occurring more frequently in cooler and more humid seasons of 2004 and 2005 than in 2006. ta b l e 5 analysis of variance for seed contamination by ascochyta blight fungi and alternaria alternata occurring on edible and fodder cultivars over cultivars, localities and years source of variation type of cultivars four edible three fodder d.f. p­value (level of significans) h. ns d.f. p­value (level of significans) h. ns fungus 3 0.0000 significant 2 3 0.0000 significant 2 locality 7 0.0000 significant 4 7 0.3694 non­significant 1 cultivar 3 0.8108 non­significant 1 2 0.2038 non­significant 1 year 2 0.5157 non­significant 1 2 0.0096 significant 2 ta b l e 6 multiple range test for ascochyta blight fungi and alternaria alternata frequency occurrence on both cultivar types independently on cultivar, locality and year species of fungi edible fodder cultivars percent (tp) h. gr percent (tp) h. gr m. pinodes 1.23 a 2.40 a p. pinodella 1.55 a 4.00 a a. pisi 4.29 a 3.69 a a. alternata 10.29 b 8.33 b ta b l e 7 multiple range test for incidence of all fungi contaminating seeds and 4 species of fungi on four edible cultivars in 8 localities (a mean from fungi, cultivars and years) locality contamination by all fungi four fungi percent (tp) h. gr (3) percent (tp) h. gr (4) sulejów 2.21 a 0 a kościelna w. 2.61 a 0.98 a b chrząstowo 2.68 a b 0 a kawęczyn 3.59 a b 3.50 a b cicibór 4.29 b 4.71 b c rarwino 6.03 c 7.61 c d krzyżewo 6.31 c 8.61 d radostowo 7.11 c 10.51 d 82 j. marcinkowska when inhabitance of cultivars over fungi, localities and years was analyzed statistically documented differences were only reported for fodder one (tab. 3). sokolik and zagłoba were less contaminated by all fungi compare to hubal (tab. 9). there were no significant differences between occurrence each of ascochyta blight fungi on seeds of both types of cultivars independently on cultivar, year and locality (tab.6) but when separately the factors were considered terno showed the highest infection in 3 out of 8 localities, reaching respectively, 20.22 percent in cicita b l e 8 multiple range test for incidence of fungi contaminating seeds of edible and fodder cultivars during years 2004-2006 (a mean from fungi, cultivars and localities) year cultivars edible fodder all fungi all fungi four species percent (tp) h. gr (2) percent (tp) h. gr (3) percent (tp) h. gr (2) 2006 3.60 a 3.11 a 2.56 a 2004 4.73 b 4.28 b 5.21 b 2005 4.73 b 5.55 c 6.09 b ta b l e 9 multiple range test for incidence of all fungi on 3 fodder cultivars (a mean from locality, year and fungal species) cultivar percent (tp) h. gr sokolik 3.80 a zagłoba 4.00 a hubal 5.14 b ta b l e 1 0 occurrence frequency of ascochyta blight fungi on seeds of edible and fodder cultivars during 2004-2006 at different localities (full names are in table 1) infection percent of seeds in different localities and years edible cultivars and their location year fodder cultivars and their location c ka kw k r rr s ch c ka b l mr m tb w 1.ramrod 1. hubal 0.44 0.22 0 6.44 0.44 2.89 0 0 2004 1.78 0.22 0 0 1.11 0 2.22 0.22 0.22 0 0 0.66 0.44 2.99 0 0 2005 0 0.22 0 2.67 1.56 0 5.78 3.33 0 0 0 0 0.22 0 0 0 2006 0 0 0 0 0 0 0 0 2.set 0.89 2.sokolik 1.33 0.89 0 17.56 5.11 0.67 0 0 2004 1.56 0 0 0 0.22 0 1.56 0 0 0.22 0 0 0 1.56 0 0 2005 0 0 0 0 1.11 0.88 2.0 1.11 0 0 0 0 0.22 0 0 0 2006 0 0 0 0 0 0 0 0 3. tarchalska 0 3.zagłoba 0.67 0.44 0 7.11 0 0.22 0 0 2004 7.99 0 0.22 0.22 1.56 0 1.11 0 0 0 0.22 0 0.22 0 0 0 2005 0 0 0 1.11 0.88 0 0.66 2.22 0 0 0 0 0 0 0 0 2006 0 0 0 0 0 0 0 0 4. terno 0.22 20.22 2,89 2.44 19.11 12.4 0 0 0 2004 0 0 0 0 0 3.12 0 0 2005 0 0.22 0 0.22 0.22 0 0 0 2006 fungi occurrence on seeds 83 bór, 19.11 in krzyżewo and 12.4 in radostowo (tab. 10). data were given as a sum of seed percent infected by m. pinodes, a. pisi and p. pinodella. the second stronger infected was set in krzyżewo (17.56) and radostowo (5.11). such high infection was only observed on seeds harvested in 2004, but those of 2006 were almost clean, since only from single ones of terno, set and ramrod fungi responsible for ascochyta blight were isolated. during three years no infection was noted on seeds from chrząstowo and sulejów, but the highest was found at krzyżewo, radostowo, cicibór and rarwino. seed infection of fodder cultivars by m. pinodes, a. pisi and p. pinodella was higher in 2004 than in 2005, and not observed in 2006 (tab. 10). in general these cultivars were less infected to edible once, reaching on zagłoba from 0.22 % in bobrowniki and lubinicko to 7.99 % in cicibór. the highest percentage (5.78) of hubal infection was noted in 2005 at tomaszów b., the locality where ascochyta blight fungi occurred in 2004 and 2005, like similarly at marianowo. these fungi were isolated only once from seeds harvested in bobrowniki and masłowice. in the period of studies (2004-2006) seed infection by fungi responsible for ascochyta blight was very low (fig. 1). a. pisi was isolated most often, independently on cultivars, years and localities totally 1.54 % of seeds, from 1.46 % in 2004 to 0.02 % in 2006 were infected. this species was the only one noted on seeds harvested in 2006. it dominated on seeds harvested in 2004, but in the next year p. pinodella was the most often isolated species. fig. 1. occurrence frequency (%) of ascochyta pisi (ap), mycosphaerella pinodes (aps) and phoma pinodella (pmp) on seeds over cultivars and localities from 2004-2006. 86 j. marcinkowska discussion occurrence frequency of fungi inhabiting pea seeds was first of all depended on species. seeds of all sample transmitted a. alternata and this species dominated among other fungi. the presented results supported the earlier obtained for field pea by filipowicz (1976) and marcinkowska (1997), green pea (czyżewska 1976) and dry edible pea (marcinkowska 1998). the next in this study were species of penicillium with p. expansum, occurring in majority, but also p. claviforme contaminated many seeds. filipowicz (1976) reported even more often penicillium sp. than a. tenuis nees (syn. a. alternata). also marcinkowska (1998) noted in 1992 common occurrence of penicillium sp. frequency of stemphylium botryosum, weak polyphagous pathogen, the third more often inhabiting species, was similar to reported earlier (grzelak, iłłakowicz 1973; marcinkowska 1997, 1998). it was important to notice that seeds were contaminated not only by penicillium spp. but also aspergillus spp., of which genera some species, like a. flavus and so p. expansum could be able to produce mycotoxins (kozakiewicz 1990, 1992). so one had to realize the presence of those species on seeds as dangerous, but on the other hand not all isolates, even of harmful species could produce mycotoxins, and their amount might be also changeable depend on different factors, like temperature or light of environment. number of isolated species differed for various studies, filipowicz (1976) reported the highest (30), the lowest (8) grzelak and iłłakowicz (1973), but in presented work were isolated 27, in 2004 slightly cooler and of higher rainfalls, and 16 in 2006 of wormer and drier vegetation season (figs 4, 5). not only the number but also composition of species inhabiting seeds was influenced by weather conditions, particularly pea pathogenic fungi for which development a drop of water is necessary to infect a plant. specific pathogens of peas, ascochyta complex fungi, were noted each season less frequently to common saprobe, a. alternata, like it happened for dry seeds in early 90.ties (marcinkowska 1998). while on fall-planted austrian winter pea, even a. alternata occurred very often, p. pinodella dominated in poland on seeds in 1994 and m. pinodes in 1993 (marcinkowska 1997), like in canada (morrall et al. 2005) and france in recent years (fougereux et al. 2006). in poland also grzelak and iłłakowicz (1973) and filipowicz (1976) reported common occurrence of fungi responsible for ascochyta blight, but with the highest for a. pisi. these earlier results concern the last species frequency were supported by the recent data from canada (morrall et al. 2005) and the presented one, even level of seeds infected by ascochyta complex fungi was very low. a. pisi, took first place and was the only one out of three, however very sporadic, in the warmest growing season of 2006. on the other hand no significant differences were found between frequency of m. pinodes, p. pinodella and a. pisi, when they were compared to a. alternata. the level of seed infection by m. pinodes and p. pinodella in case of majority of species inhabiting was similar for both groups of cultivars, and lower comparing to a. pisi. the last species infected significantly more seeds of edible cultivars to fodder one. even more to f. poae, the saprobic species of fusarium inhabiting pea seeds most often. this domination was already proved by filipowicz (1976) and marcinkowska (1993, 1997, 1998). the number of isolated fusarium species was lower to obtained by filipowicz (1976) but the same as marcinkowska (1993, 1997), however the prevalence of species varied between reports. among pathogenic species czyżewska (1976) and filipowicz (1976) fungi occurrence on seeds 87 noted more often f. oxysporum. in this study was also found f. solani reported earlier on seeds of winter pea (marcinkowska 1997). variability of mycobiota was influenced, besides fungal species and weather conditions of the growing season, also by characters of cultivars and environmental conditions of localities. the last factor was combined with local weather or even microclimate of field where peas were growing in different years. particularly rainfalls increased frequency of fungi inhabiting seeds, so for locality characterized by higher precipitation and usually lower temperatures (figs. 4 and 5) many more fungi were isolated. the influence of weather was also reported from other studies (bathgate et al. 1989; xue et al. 1997; roger et al. 1999; marcinkowska 1998, 2002; morrall et al. 2005; fougereux et al. 2006). presented data supported positive (2005 and 2004) or negative (2006) influence of weather on seed infection by plant pathogens but also their inhabitance by saprobic species, especially on cultivars suitable for light soil. influence of cultivars on fungal species occurrence was proved by several authors (filipowicz 1993; fougeroux et al. 2006; marcinkowska 1997, 1998, 2002; xue et al. 1996). these data reported only such dependence considered all fungi on fodder cultivars. the obtained results supported the earlier reports cited here, done by different researches in various countries, that incidence of ascochyta blight fungi was influenced by many factors, changeable in between studies. according to fougereux et al. (2006) and morrall et al. (2006) these were mainly years, production area (localities) and type of crop (winter or spring pea). conclusions 1. many more seeds were inhabited by saprobic fungi than plant pathogenic. number of fungi occurring on edible and fodder cultivars depended mainly on fungal species and year. 2. alternaria alternate was the most often occurring species on both types of cultivars, next was penicillium spp. 3. stemphylium botryosum, the weak pathogen, was isolated most often from seeds of both cultivar types compare to all other pathogenic fungi. 4. fungi responsible for ascochyta blight infected seeds sporadically. they were recovered with various intensity from seeds of both types of cultivars. ascochyta pisi was most often isolated. 5. no clear response of tested pea cultivars to ascochyta blight fungi occurrence on their seeds was found since host­plant reaction was strongly influenced by environmental conditions (locality and year). 6. differences in seed contamination by any fungi species were only found for fodder cultivars. references bathgate j. a., sivasithamparam k., khan t. n. 1989. identity and recovery of seed-borne fungal pathogens of field peas in western australia. n. zeal. j. crop hort. sci. 17: 97–101. bretag t. w., keane p .j., price t. v. 1995. effect of ascochyta blight on the grain yield of field peas (pisum sativum l.) grown in southern australia. austr. j. exp. agric. 35: 531–536. czyżewska s. 1976. studies on fungi infecting seeds of pea (pisum sativum l.). biul. warz. 19: 271–288. filipowicz a. j. 1976. badania mikoflory nasion grochu siewnego (pisum sativum l.) ze szczególnym uwzględnieniem grzybów z rodzaju ascochyta i fusarium. roczn. n. roln., seria e, 5 (2): 85–120. 88 j. marcinkowska filipowicz a. j. 1993. podatność odmian i rodów grochu (pisum sativum l.) o nasionach żółtych i gładkich na grzyby patogeniczne. biul. ihar 186: 89–94. fougereux j. a., mériaux b., olivier v., sérandat i., leclerc s., avrillon m., cassignol f., dagorn c. 2006. a 20 years overview of pea seed contamination by ascochyta sp. in france. poster abstract (c11), ascochyta workshop on grain legumes in le tronchet (brittany, france) on 2-6 july, 2006. grzelak k., iłłakowicz a. 1973. grzyby z rodzaju ascochyta w laboratoryjnej ocenie zdrowotności nasion grochu (pisum sativum l.). biul. ihar 3/4: 155–161. kozakiewicz z. 1990. aspergillus spp. descriptions of pathogenic fungi and bacteria 100: 991–998. kozakiewicz z. 1992. penicillium spp. descriptions of pathogenic fungi and bacteria111:1101–1110. marcinkowska j. z. 1993. fusarium spp. as pathogens of dry peas (pisum sativum l.) in poland. hod. rośl. aklim. nasien. 37 (4): 95–102. marcinkowska j .z. 1997. micromycetes on pisum sativum var. arvense. acta mycol. 32 (1): 31–39. marcinkowska j. z. 1998. variability of dry seed mycobiota of pisum sativum. acta mycol. 33 (1): 91–99. marcinkowska j. z. 2002. foliar diseases of pisum sativum l. in poland. pl. breed. seed sci. 46 (1): 49–54. marcinkowska j. z. 2003. oznaczanie rodzajów grzybów ważnych w patologii roślin. fundacja rozwój sggw, warszawa, 328pp. marcinkowska j., schollenberger m. 1979. mikroflora nasion soi. roczn. nauk roln., seria e, 9 (2): 41–54. morrall r. a. a., carriere b., ernst b., pearse c., schmeling d., thomson l. 2005. seed-borne pathogens of pea in saskatchewan in 2004. can. plant dis. surv. 85: 91–93. morrall r. a. a., carriere b., ernst b., pearse c., schmeling d., thomson l. 2006. seed-borne pathogens of pea in saskatchewan in 2005. can. plant dis. surv. 86: 109–111. nowicki b. 1997. patogeny pietruszki korzeniowej występujące na nasionach. acta agrobot. 50 (1/2): 27–34. roger c., tivoli b., huber l. 1999. effects of temperature and moisture on disease and fruitbody development of mycosphaerella pinodes on pea (pisum sativum). plant path. 48:1–9. skolko a. j., groves j. w., wellen v. r. 1954. ascochyta diseases of peas in canada with special reference to seed transmission. can. j. agric. sci. 34 (4): 417–428. xue a. g., warkentin t. d., greeniaus m. t., zimmer r. c. 1996. genotypic variability in seedborne infection of field pea by mycosphaerella pinodes and its relation to foliar disease severity. can. j. plant path. 18: 370–374. xue a. g., warkentin t. d., kenaschuk e. o. 1997. effects of timings of inoculation with mycosphaerella pinodes on yield and seed infection of field pea. can. j. plant sci. 77: 685–689. występowanie grzybów na nasionach grochu polnego s t r e s z c z e n i e zasiedlenie nasion 7 odmian grochu polnego przez różne gatunki grzybów oceniano w latach 2004-2006. odmiany polskie: ramrod, set i tarchalska, oraz czeska – terno, wymagają do uprawy gleb żyźniejszych w porównaniu do paszowych: hubala, sokolika i zagłoby, odpowiednich do uprawy na glebach lżejszych. stąd każda z grup odmian uprawiana była w 8 miejscowościach, reprezentujących rejony uprawy grochu w polsce, przy tym w 6 różnych, jedynie w ciciborze i kawęczynie były obydwa doświadczenia. ze 168 próbek, każda 450 nasion, odkażonych powierzchniowo wyizolowano w teście szalkowym na pożywce coon’a (cn), 27 gatunków w 2004, 22 w 2005 oraz 16 w 2006 roku. alternaria alternata uzyskano z nasion wszystkich próbek, w największej liczbie. kolejnym był stemphylium botryosum, słaby patogen, gatunki rodzaju penicillium, a zwłaszcza p. expansum, ascochyta pisi, fusarium poae, cladosporium spp. z gatunków chorobotwórczych najrzadziej infekował botrytis cinerea, rhizoctonia solani, f. solani, sclerotinia sclerotiorum. sprawcy zgorzelowej plamistości grochu, zwłaszcza mycosphaerella pinodes (0.36%) i phoma pinodella (0.66%) zainfekowały nieliczne nasiona, przy tym tylko a. pisi wystąpił we wszystkie 3 lata, osiągając średni najwyższy procent fungi occurrence on seeds 89 porażenia (1.54) bez względu na odmiany i miejscowości. te trzy gatunki w sumie porażały najsilniej odmianę terno, zasiedlając najliczniej nasiona z krzyżewa, radostowa, ciciboru i rarwina. w chrząstowie i sulejowie w ogóle nie zanotowano sprawców askochytozy. z odmian uprawianych na glebach lżejszych, zagłoba wykazała najsilniejsze porażenie. najwięcej sprawców zgorzelowej plamistości grochu izolowano z nasion zebranych w tomaszowie bolesławickim i marianowie. grzyby te nie poraziły nasion w 2006, najbardziej suchym i gorącym w okresie badań. częstotliwość występowania grzybów zależała nie tylko od ich gatunku, co udowodniono statystycznie, ale w znacznej mierze od warunków otoczenia, na które wpływały warunki atmosferyczne pola uprawnego, a więc wskazane wyżej różnice dla miejscowości i lat, również często istotne. najsilniej zasiedlone przez grzyby były nasiona zebrane w roku 2004, jak również w tym sezonie gatunki wystąpiły najliczniej. cechy odmian, udowodnione tylko dla trzech o mniejszych wymaganiach glebowych, nie pozostawały bez wpływu na występowanie grzybów. uzyskane wyniki wskazały, iż różne czynniki, ale w niejednakowym stopniu, zmieniały zasiedlenie nasion przez gatunki grzybów, tak porażających je jak i jedynie zanieczyszczających. 2014-01-01t11:47:25+0100 polish botanical society urocystis skirgielloi, a new graminicolus smut fungus infecting heteropogon contortus in india marcin piątek department of mycology, w. szafer institute of botany, polish academy of sciences, lubicz 46, pl 31 512 kraków, mpiatek@ib pan.krakow.pl p i ą t e k m.: urocystis skirgielloi, a new graminicolus smut fungus infecting heteropogon contortus in india. acta mycol. 41(1): 7 10, 2006. urocystis skirgielloi m. piątek sp. nov. is described and illustrated on the basis of infected leaves of heteropogon contortus (l.) roem. & schult. collected in india. this is the first urocystis species on the grass genus heteropogon as well as on any grass genera belonging to the subtribe anthistiriinae of the tribe andropogoneae (poaceae). key words: urocystis, urocystaceae, urocystales, ustilaginomycetes, india, asia introduction in respect to smut fungi, india is one of the best-explored countries in the asiatic mainland thanks to such giants of indian taxonomy as b. b. mundkur, m. j. thirumalachar, m. s. pavgi, and many others, whose research has been supplemented by excellent works of k. vánky, a hungarian smut taxonomist. however, most reports, especially those published in regional indian journals, are scattered in a number of papers that are often not easily accessible to smut taxonomists. a monograph study of smut fungi of india would be enthusiastically welcomed by the world’s scientific community. although well studied, india is still a paradise for mycologists, and new smut fungi are described from the area nearly every year. another new species was found in unidentified materials obtained from the imi herbarium, infecting leaves of heteropogon contortus (l.) roem. & schult., in kashmir. it belongs to the genus urocystis rabenh. ex fuckel, and is described and illustrated below. measurements of spores were taken by standard light microscopy (lm) in lactophenol heated to the boiling point and then cooled. the sizes given below are the means of 30 measurements of spore balls, spores and sterile cells. the sem studies proceeded as described by pi ą t e k et al. (2005). it is great pleasure to dedicate this fungus to professor alina skirgiełło, at the occasion of her 95 birthday. it is tribute to her many-sided mycological activity and arranging the professional mycological studies in poland. acta mycologica vol. 41 (1): 7-10 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday urocystis skirgielloi 9 composed of 1-4(-7) central spores (1 – 7.8%, 2 – 32.2%, 3 – 35.6%, 4 – 20.0%, 5 – 2.2%, 6 – 0.5%, 7 – 1.7%), surrounded by usually continuous layer of sterile cells, rarely the layer is not complete. – spores globose, subglobose, ellipsoidal or slightly irregular, olive-brown or reddish-brown, (11-)12-14(-16) × 9-12(-13) μm, smooth in lm and sem. – sterile cells globose, subglobose, ellipsoidal or irregular, usually with flattened contact sides, yellowish, (5-)6-11 × (4-)5-8(-9) μm, wall ca 1 μm thick, smooth in lm and sem. host and distribution. on poaceae: heteropogon contortus (l.) roem. & schult.; india, asia. known only from the type locality. urocystis skirgielloi is the first species of urocystis on the genus heteropogon pers. (poaceae). this is a small grass genus, belonging to the subtribe anthistiriinae of the tribe andropogoneae (c l a y t o n , re n v o i z e 1986), with six species occurring mostly in tropical and subtropical areas. heteropogon contortus, which is the host plant of u. skirgielloi, is the most common member of the genus, widely distributed in tropical, subtropical and warm-temperate regions of africa, asia, europe, north and south america, and australia (c l a y t o n et al. 2006). the host plant in the type collection is sterile and does not contain any inflorescence. the comparison of this material with the specimens of heteropogon contortus preserved in the phanerogamic herbarium of the w. szafer institute of botany in kraków (kram) shows that they are morphologically similar. v á n k y (2000) revised smut fungi infecting heteropogon and recognized eight species. however, all of them are completely different from urocystis skirgielloi as they belong to the genus sporisorium ehrenb. ex link, characterized by sori with peridium and columellae, spores usually arranged in spore-balls, and the presence or absence of sterile cells. all sporisorium species known on heteropogon produce sori in inflorescences of host plants. urocystis skirgielloi has also no comparable species on other grasses closely related to heteropogon. of the 12 genera included in the subtribe anthistiriinae by c l a y t o n and re n v o i z e (1986), no urocystis species had been described previously. acknowledgements. i am grateful to my wife dr. jolanta piątek (kraków, poland) for her illustrations, to the curator of imi for the loan of the specimen of the new urocystis species, and to anna łatkiewicz (kraków, poland) for her assistance with sem pictures. this study was supported by the polish ministry of education and science for the years 2005 2008, grant no. 2 p04g 019 28. references c l a y t o n w. d., re n v o i z e s. a. 1986. genera graminum. grasses of the world. kew bull., additional series 13: 1 389. c l a y t o n w. d., h a r m a n k. t., wi l l i a m s o n h. 2006. world grass species: descriptions, identifi cation, and information retrieval. http://www.kew.org/data/grasses db.html [accessed 20 february 2006]. p i ą t e k m., ru s z k i e w i c z m i c h a l s k a m., m u ł e n k o w. 2005. catalogue of polish smut fungi, with notes on four species of anthracoidea. polish bot. j. 50(1): 19 37. v á n k y k. 2000. taxonomical studies on ustilaginales. xx. mycotaxon 74 (1): 161 215. 10 m. piątek urocystis skirgielloi, nowy gatunek głowni porażający heteropogon contortus w indiach s t r e s z c z e n i e w pracy opisano i zilustrowano nowy gatunek głowni, urocystis skirgielloi m. piątek sp. nov., porażający liście heteropogon contortus (l.) roem. & schult. w indiach. jest to pierwszy gatunek z rodzaju urocystis stwierdzony na trawach z rodzaju heteropogon jak również na 11 pokrewnych rodzajach traw należących do podplemienia anthistiriinae i plemienia andropo goneae (poaceae). 2014-01-01t11:43:05+0100 polish botanical society robillarda sessilis, a rare coelomycete isolated from scots pine seedlings eugene yurchenko and dasha belomesyatseva laboratory of mycology, v.f. kuprevich institute of experimental botany akademichnaya street 27, by-220072 minsk, eugene_yu@tut.by yurchenko e., belomesyatseva d.: robillarda sessilis, a rare coelomycete isolated from scots pine seedlings. acta mycol. 45 (1): 27–32, 2010. a coelomycete with appendage-bearing conidia, r. sessilis, was isolated three times from stems of living healthy pinus sylvestris seedlings of the 1st year growing in a nursery in central belarus. macroscopic and microscopic morphology of the fungus in culture is described. key words: belarus, nursery, pinus sylvestris, stem introduction robillarda sessilis (sacc.) sacc. was found during studies of the fungi inhabiting stems of healthy pinus sylvestris seedlings in a sample nursery in central belarus. this species is a remarkable fungus with conidia bearing three long hair-like appendages. it was selected as a generic type for robillarda, an anamorphic genus with unknown teleomorph (nag raj 1993). altogether 35 species were published under this generic epithet, 12 of which were referred, according to nag raj (1993), to seven other genera. material and methods the inventory of the fungi was carried out by sampling plants in 1 m2 plots on a 800 m2 plantation situated in central belarus. a first sample (taken 2 vi 2009) included 5 well-developed seedlings from a single plot located in the center of plantation. a second sample (taken 22 vi 2009) included 25 equally well-developed seedlings collected from 5 plots (5 plants per plot). four plots were situated near corners of acta mycologica vol. 45 (1): 27–32 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 28 e. yurchenko and d. belomesyatseva the plantation, one plot in the plantation center (this central plot position did not coincide with the plot in previous sampling). for isolation of the fungi, the seedlings were washed 3 min under a strong stream of tap water on a sieve. each seedling stem, from the root neck to the foot of leaf whorl, was cut using a sterile razor blade into ca 2 mm long segments. the segments were put on malt extract agar (mea: 1% malt extract, amresco, usa, and 1.5% agar) with addition of tetracycline 30 μg/ml (amresco), and incubated for 10 days at 26°c. for the control that the propagules did not originate from tap water, 1.5 ml of the water was poured in petri dish with solid 2% mea, in 3 replicates. mycelia growing from stem pieces and producing pycnidia, were transferred for storing on mea slants under mineral oil. for describing cultural morphology, isolates were taken from storage and cultured on mea (2% malt extract, 1.5% agar) at 26°c in the dark. for describing micromorphology, preparations were mounted in 3% koh water solution. dry cultures were deposited in v.f. kuprevich institute of experimental botany herbarium (msk-f). results it the first sample from the plantation center (2 vi 2009), one isolate of r. sessilis was obtained. in the second sample (22 vi 2009) two isolates were obtained from 2 of 25 studied plants, one from central, one from a corner plot. the morphological diagnosis of r. sessilis, based on cultures, is given below. morphological description. after 1 week: mat 25 mm in diam, more or less rounded, colorless, low felty, near 1 mm high, more or less fasciculate or tufted in periphery zone and just near the point of inoculation; margin somewhat wavy, abrupt, with scarce cilia-shaped hyphae; reverse pale apricot yellow. after 2 weeks: mat about 50 mm in diam, very pale pinkish cream, in central and middle zone floccose-felty; periphery zone clearly delimited, ca 10 mm wide, low velvety, with slight radial depressions; margin without aerial growth, transparent; reverse in central and middle zone brownish yellow with orange tint, with blackish or dark olive spots due to the presence of conidiomata. conidiomata occurring about 1.5–2 weeks post inoculation, especially developing when the medium dries up, in mat periphery and middle zone scattered, but at the border between these zones abundant and often aggregated, forming a ring, having dark brown color in reverse (fig. 1). aerial hyphae moderately branched, somewhat wavy, but with even walls, smooth, hyaline or subhyaline, narrower ones (2-3.5 μm) long-celled and small-guttulate, wider ones (3.5-7.7 μm) with short swollen cells and rich-guttulate; thin lateral branches 0.8-1 μm wide, tapering. repent and immersed hyphae moderately branched, with short more or less swollen cells, rich-guttulate, often disintegrating at septa, 1.7-8.7 μm wide. marginal hyphae rather richly branched, divided into axial hyphae and their lateral branches; axial hyphae 4.3-8.2 μm wide, with short, more or less swollen, somewhat guttulate cells; lateral branches 3-4 μm wide, having blunt apices 2-3.2 μm wide (fig. 2). fig. 1. two-weeks old culture of robillarda sessilis on 2% mea: face of the mat (above) and reverse (below). scale bar = 1 cm. robillarda sessilis 29 conidiomata pycnidioid, solitary, 100-150(-500) μm in diam, or aggregated 3-10 and more and partly confluent (aggregations to 1-1.5 mm in diam), globose or irregular, flattened when large, fully or partly immersed, black, glabrous, glossy, with uneven surface, ostioles not visible; conidiomatal wall ca 25-35 μm thick, of pseudoparenchymatic cells, outer layer dark brown, 7-10 μm thick, mainly of textura angularis (partly also of narrow textura prismatica) with somewhat thick-walled cells 4-13(-17.5)×(1.5-)2.2-7 μm, turning into a hyaline inner layer of thin-walled and guttulate cells 7-30×2.5-5 μm. conidiophores reduced, mostly one-celled, in shape of somewhat elongated conidiogeneous cells, originating from pseudoparenchyma lining the inner surface of the locule and indistinctly differring from pseudoparenchymatic cells. conidiogeneous cells ampulliform to subcylindrical, hyaline, thin-walled, smooth, 6-11×2-3 μm. conidiogenesis holoblastic with limited sympodial proliferation (sometimes two conidia simultaneously developing on the same cell); conidia originating at apices of conidiogeneous cells, often sitting on very small protuberances or denticles, at early developmental stages without appendages; in a layer with many developing conidia, appendages situated parallel or divergent at a very sharp angle. conidia 2-septate, consisting of a two-celled conidium body and an apical cell with appendages; conidium body medianly 1-euseptate, fusiform, straight or a little curved, often very little constricted at the septum, basally more or less truncated, subhyaline to pale brown, usually yellowish, smooth, thin-walled, 9.3-12(-14.2)×2.7-3.5 μm; apical cell hyaline, very thin-walled, short cylindrical basally for 1-2 μm, divided into 3 hyaline, smooth, hair-like, divergent, straight or curved, more or less apically attenuated appendages 17-26 μm long and 0.5-0.6 μm wide in the middle part; older conidia in preparation often loosing apical cell (fig. 3). substrata and general distribution. r. sessilis was documented from quite variable hosts and substratum types: on stems (bark), dead branches, leaves (causing leaf spot), seeds of bischofia, cocos, ficus, fragaria, fumana, ludwigia, magnolia, paeonia, quercus, randia, rosa, rubus, vitis. the fungus has been recorded in europe (hungary, italy), asia (india), north america (usa), caribbeans, africa (angola), and on the plant material imported from japan. the majority of records are from india (nag raj 1993). in 1970 this species was described from northea seeds imported to st-petersburg, russia, from indonesia, under the name mycohypallage northeae (melnik 1970). gasich (1995) collected this fungus in saratov oblast, russia, where it caused leaf spot disease of eryngium (see also melnik 1997). so far, five localities, including the type locality in italy and our find, but excluding the find on imported seeds, are know from europe today. in nebraska, usa, r. sessilis was collected on pinus ponderosa, but details about the age of the plants or the kind of organs are unknown (nag raj 1993). the fungus was also isolated from soil in australia (matsushima 1989) and from a soil sample collected in mountain pine forest in pakistan (matsushima 1993). our case indicates that r. sessilis is capable of infecting stem tissues of healthy, vigorous pinus sylvestris seedlings as either an endophyte or phylloplane component. the fungus can be named more widely ‘associated with seedling’ because of used method of sterilization. conidiomata were not observed on this host. only three plants colonized by the fungus, were detected among 30 ones subjected to cultural experiment, and thus the species can be regarded as rare in the studied plantation. 30 e. yurchenko and d. belomesyatseva fig. 2. robillarda sessilis hyphae in culture: a – aerial, b – repent and immersed, c – marginal. scale bar = 10 μm. robillarda sessilis 31 material examined: belarus, minsk oblast, dzyarzhynsk district, near enerhetykau settlement, basic forest nursery of belarusian state technological university, plantation of 1st year pinus sylvestris, coll. e. yurchenko 2 vi 2009 (msk 7065); ibid., coll. e. yurchenko 22 vi 2009 (msk 7138a, b). acknowledgements. the authors are grateful to dr g.j.m. verkley (fungal biodiversity centre, centraalbureau voor schimmelcultures, utrecht, the netherlands) for critical consideration of the ms. this work is a part of the study supported financially by grant b09-126 of belarusian republican foundation for fundamental research. fig. 3. robillarda sessilis: a – conidiophores and developing conidia, b – mature conidia with appendages, c – conidia that lost apical cell msk 7065. scale bar = 10 μm. 32 e. yurchenko and d. belomesyatseva references gasich e. l. 1995. robillarda sessilis (sacc.) sacc. – the first record in russia. mikologiya i fitopatologiya 29 (5/6): 16–18. matsushima t. 1989. matsushima mycological memoirs no. 6. (in:) matsushima mycological memoirs no. 01 (1980) – no. 10 (2001). electronic version. september 15, 2005. matsushima t. 1993. list of microfungi from pakistan soils. (in:) t. nakaike, s. malik (eds). cryptogamic flora of pakistan 2: 43–63. national science museum, tokyo. melnik v. a. 1970. species sphaeropsidalium novae et curiosae. nov. syst. pl. non vasc. 7: 238–242. melnik v. a. 1997. definitorium fungorum rossiae. classis coelomycetes. fasc. 1. genera rara et minus cognita. nauka, petropoli, 280 pp. nag raj t. r. 1993. coelomycetous anamorphs with appendage-bearing conidia. mycologue publications, waterloo, 1101 pp. 2014-01-01t11:50:10+0100 polish botanical society hypogeous fungi of lithuania: a preliminary checklist ernestas kutorga1,2 and marija kataržytė1 1department of botany and genetics, vilnius university m.k. čiurlionio street 21/27, lt-03101 vilnius, ernestas.kutorga@gf.vu.lt 2laboratory of mycology, institute of botany, žaliųjų ežerų street 49, lt-08406 vilnius kutorga e., kataržytė m.: hypogeous fungi of lithuania: a preliminary checklist. acta mycol. 43 (2): 133–138, 2008. the paper reports on hypogeous fungi known from lithuania, and data on their habitats, phenology, and distribution. references on the collections kept in the herbaria are also pointed out. the information is based on literature data and re-examination of all available voucher specimens. 22 species (12 genera, 3 phyla) recorded from 124 localities are presented in a preliminary checklist. key words: hypogeous fungi, diversity, distribution, lithuania introduction hypogeous fungi of lithuania are poorly known as only few mycologists have investigated them. the first relevant data on six species of hypogeous fungi were published by bucholtz (1904, 1907), one of the pioneers in the study of sequestrate fungi (castellano et al. 2004). f. bucholtz’s examined specimens collected by m. domaradsky from kaunas surroundings in 1904. unfortunately, all lithuanian collections seen by f. bucholtz were lost during first world war. some records of hypogeous species were published by trzebiński (1934), mazelaitis and minkevičius (1957), mazelaitis (1961, 1966). these species belonging to leucogaster r. hesse, melanogaster corda and rhizopogon fr. & nordholm were included by mazelaitis (1982) in his monograph on lithuanian gasteromycetes. the data on ascomycete species of choiromyces vittad., geopora harkn., hydnotrya berk. & broome and tuber f. h. wigg. have been published in a monographic work on the pezizales of lithuania (kutorga 2000). recently undertaken special sampling by raking the soil has extended the known range of several species, and added three species to the lithuanian mycobiota (kataržytė, kutorga 2005). the aim of the present study is to re-examine all available material on hypogeous fungi known in lithuania, to re-evaluate taxonomic arrangement and nomenclature for all taxa treated, and finally to present an annotated checklist of these fungi. acta mycologica vol. 43 (2): 133–138 2008 134 e. kutorga and m. kataržytė materials and methods all relevant specimens in fungal reference collections of the institute of botany, vilnius (bilas) and vilnius university (wi) have been located and re-examined. the checklist includes also all the published records of species from lithuania. the concept of hypogeous fungi follows castellano et al. (2004). other sequestrate fungi that produce sporocarps above soil surface in lithuania, i.e. from the genera scleroderma pers. and endoptychum czern., are not included in this checklist. for each fungus the information is given in the following order: scientific name, it’s synonym (when used in lithuanian literature), data on habitat and phenology: months in which fungus has been observed/collected), distribution (distr: number of known localities and administrative districts/cities); preserved and examined fungal exsiccata (exs.; an acronym of herbarium in which specimens are deposited, and herbarium access number, fungal name on the label, if current identification is different), literature (lit.), and notes (when necessary). taxa of glomeromycota, ascomycota, and basidiomycota are treated separately in alphabetical order. the taxonomical arrangement of taxa follows kirk et al. (2008), and an author citation was used according the authors of fungal names (2 ed., http://www.indexfungorum. org/ authorsoffungalnames.htm). list of taxa glomeromycota endogone lactiflua berk. & broome under picea abies, in ca. 90 years old norway spruce wood, october. distr.: 1 locality, plungė district. exs.: wi (5476). lit.: kataržytė and kutorga (2005). ascomycota choiromyces meandriformis vittad. (=choiromyces venosus (fr.) th. fr.) in the vicinity of deciduous trees (quercus robur, betula sp., alnus sp., salix sp.), in deciduous, mixed and coniferous woods, august–october. distr.: 4 localities (documented by specimens), ignalina, marijampolė, molėtai and raseiniai districts; additional 4 localities (not documented), kėdainiai, klaipėda, molėtai and panevėžys districts. exs.: bilas (4898, 9643, 18624), wi (3350). lit.: mazelaitis and minkevičius (1957), mazelaitis (1966), ławrynowicz (1990), kutorga (2000). elaphomyces asperulus vittad. under pinus sylvestris and picea abies, in coniferous woods, occasionally in mixed woods with betula, april–june, august–october. distr.: 28 localities, anykščiai, elektrėnai, kupiškis, panevėžys, plungė, �alčininkai, �ilutė, tauragė, varėna, vil-škis, panevėžys, plungė, �alčininkai, �ilutė, tauragė, varėna, vil-panevėžys, plungė, �alčininkai, �ilutė, tauragė, varėna, vilnius and zarasai districts, and neringa. exs.: bilas (3517 as elaphomyces cervinus, 7582 as e. muricatus, 14066 as e granulatus, 14416 as e. cervinus, 18633, 18635, 19440, 21247 all latter four as e. granulatus, 24544, 24747, 28928), wi (5453, 5454, 5465–5472, 5551); in collections of cordyceps ophioglossoides – bilas (2918, 2981, 2988, 3295, 3931), wi (3005). lit.: kataržytė and kutorga (2005). hypogeous fungi of lithuania 135 notes. e. asperulus is distinguished from e. granulatus mainly on the basis of spore ornamentation and colour of peridium (ławrynowicz 1988). e. asperulus has spore spines up to 2 µm long and whitish to greyish pink peridium. e. granulatus is characterised by spines longer than 2 µm and whitish to yellowish peridium. elaphomyces granulatus fr. (=elaphomyces cervinus (pers.) schltdl.) mostly under pinus sylvestris and picea abies, rarely associated with quercus robur, in coniferous and mixed woods, may–october. distr.: 14 localities, plungė, �akiai, tauragė, varėna and vilnius districts. exs.: bilas (4875 as elaphomyces variegatus, 18632, 24545), wi (5452, 5455, 5458–5464, 5474, 5552). lit.: mazelaitis (1966), kataržytė and kutorga (2005). elaphomyces muricatus fr. (=elaphomyces variegatus vittad.) under picea abies and pinus sylvestris, in coniferous woods, occasionally in mixed woods with betula and populus tremula, april, july–september. distr.: 11 localities, alytus, anykščiai, jurbarkas, kretinga, plungė, �irvintos and tauragė districts. exs.: bilas (18634, 18781, 19411 all as e. granulatus), wi (5475); in collections of cordyceps ophioglossoides – bilas (2899, 2909, 3196, 3931, 5631, 11570, 24732). lit.: bucholtz (1904), mazelaitis (1966), kataržytė and kutorga (2005). notes. the fruit-bodies of elaphomyces species were commonly observed in the places disturbed by feeding wild animals or in association of mycoparasitic species of the genus cordyceps (fr.) link. geopora arenicola (lév.) kers on soil, in coniferous and deciduous woods, usually on forest paths and in other naturally or artificially disturbed sites, occasionally in fire places, june–october. distr.: 16 localities, jonava, jurbarkas, pakruojis, plungė, �akiai, �ilutė, ukmergė, varėna and vilnius districts, and neringa. exs.: bilas (21294, 34521–34528), wi (4810, 5456, 5523–5526). lit.: kutorga (1994, 2000). hydnotrya tulasnei berk. & broome in picea abies wood, july. distr.: 1 locality, �ilutė district. exs.: bilas (18627). lit.: kutorga (2000). peziza ammophila durieu & mont. in the vicinity of grasses (ammophila arenaria, festuca arenaria, leymus arenarius), in littoral sand dunes, september–october. distr.: 4 localities, neringa. exs.: bilas (18566, 18593, 21780), wi (5527, 5528). lit.: kutorga (2000, 2004). tuber exiguum r. hesse in mixed wood, august. distr.: 1 locality, kaunas surroundings. exs.: none. lit.: bucholtz (1904), kutorga (2000). tuber puberulum berk. & broome under pinus sylvestris, in a ca. 30 years old pinus sylvestris plantation with intermixed younger picea abies trees, september–october. distr.: 1 locality, plungė district. exs.: wi (5475). lit.: kataržytė and kutorga (2005). tuber rufum pico : fr. f. nitidum (vittad.) montecchi & lazzari (=tuber nitidum vittad.) in mixed wood on slope along nemunas river, august. distr.: 1 locality, kaunas surroundings. exs.: none. lit.: bucholtz (1904), kutorga (2000). 136 e. kutorga and m. kataržytė basidiomycota hymenogaster arenarius tul. & c. tul. in mixed woods, august–september. distr.: 2 localities, kaunas surroundings. exs.: none. lit.: bucholtz (1907). notes. the specimen, reported as hymenogaster rehsteineri bucholtz (bucholtz 1904), was later reconsidered as h. arenarius by bucholtz (1907). hymenogaster citrinus vittad. in wood, august. distr.: 1 locality, kaunas surroundings. exs.: none. lit.: bucholtz (1904, 1907). hymenogaster olivaceus vittad. in a ca. 40 years old pinus sylvestris plantation with intermixed picea abies trees, august. distr.: 1 locality, plungė district. exs.: wi (5477). lit.: kataržytė and kutorga (2005). leucogaster nudus (hazsl.) hollós (=leucogaster floccosus r. hesse) not reported. distr.: 1 locality, vilnius surroundings. exs.: none. lit.: mazelaitis (1982). notes. mazelaitis (1982) noted that the species usually grows in deciduous and coniferous forests, especially in quercus stands, however, an exact data on ecology of lithuanian record was not provided. melanogaster ambiguus (vittad.) tul. & c. tul. in quercus robur wood, september. distr.: 2 localities, plungė (documented by specimen) and varėna districts (not documented). exs.: bilas (18989). lit.: mazelaitis (1982). melanogaster variegatus (vittad.) tul. & c. tul. in deciduous woods, the months of observations were not reported. distr.: 2 localities, vilnius surroundings. exs.: none. lit.: trzebiński (1934), mowszowicz (1957), mazelaitis (1982). notes. reports on m. variegatus were based on collections made in 1928 and 1952. however, there are no specimens available under this name in wi and bilas. according to the description provided by mazelaitis (1982) the basidiospores were ellipsoid, 6–9 × 3–4 µm. mentioned spore size fits better the concept of m. broomeianus berk. extremely closely related m. variegatus s. str., a species of southern and central europe, differs mainly in broader basidiospores, 7.5–10 × 5.5–8 µm (lange 1956; pegler et al. 1993). pompholyx sapida corda under tilia sp., in park, august. distr.: 1 locality, kaunas surroundings. exs.: none. lit.: bucholtz (1904). rhizopogon angustisepta zeller & c. w. dodge not reported. distr.: not reported. exs.: none. lit.: mazelaitis (1982). notes. in the treatment of gasteromycetes from the former soviet union (sosin 1973) the species was reported as known from lithuania. however, the attempt to reveal more data concerning this report has failed (mazelaitis 1982). hypogeous fungi of lithuania 137 rhizopogon luteolus fr. & nordholm in pinus sylvestris woods, july–october. distr.: 16 localities, kretinga, prienai, �alčininkai, �venčionys, tauragė and varėna districts, druskininkai and neringa. exs.: bilas (3361, 4869, 5175, 6525, 7832, 8097, 9994, 11271, 12674 as rhizopogon virens, 12709 as r. vulgaris, 12719, 12759, 13435, 20507 as r. obtextus), wi (5478). lit.: mazelaitis (1961, 1982). rhizopogon roseolus (corda) th. m. fr. in pinus sylvestris woods, july, september–october. distr.: 6 localities, ignalina, �venčionys, varėna, vilnius (documented by specimens) and molėtai (not documented) districts. exs.: bilas (13149 as rhizopogon virens, 13482, 13488, 14652), m (1809 as r. virens). lit.: mazelaitis (1961, 1982), martín (2001). notes. both r. roseolus and r. vulgaris are characterized by peridium discoloring red to purplish red on bruising, however, these taxa can be distinguished by spore size. following the concepts of taxa by pegler et al. (1993) we assigned the specimens with larger spores (7–10 ×3–3.5 µm) to r. roseolus, and those with smaller spores (5,5–7 × 2–3 μm) – to r. vulgaris. the specimen, which has been collected by g. bresadola in 1921 from varėna district and currently preserved at m, was re-examined by martín (2001). rhizopogon vulgaris (vittad.) m. lange in pinus sylvestris woods, august–october. distr.: 5 localities, varėna districts, druskininkai and neringa (documented by specimens) and molėtai (not documented) district. exs.: bilas (13087, 15357 as rhizopogon roseolus, 15238 as r. roseolus; 21218 as r. roseolus). lit.: mazelaitis (1982). discussion 22 species (12 genera, 3 phyla) recorded from 124 localities are presented in the list of hypogeous fungi from lithuania. elaphomyces asperulus, e. granulatus, e. muricatus, geopora arenicola and rhizopogon luteolus are the most commonly observed and collected species. 82 out of total 105 preserved specimens of hypogeous fungi belong to the five latter species. 13 species are known in lithuania from one or two localities only. voucher specimens of 8 species are no longer extant, and these taxa are currently known only from the literature. many more hypogeous fungi remain to be discovered in lithuania, and the knowledge of already known and yet not recorded species ecology and distribution needs to be investigated in greater depth. acknowledgements. we wish to express our gratitude to prof. j. m. trappe (u.s.a.) and dr. m. p. martín (spain) for valuable consultations during this study. we are indebted to anonymous reviewer for suggestions on the manuscript. references bucholtz f. 1904. nachträgliche bemerkungen zur verbreitung der fungi hypogaei in russland. bull. soc. imp. nat. moscou 18 (4): 335–343. bucholtz f. 1907. zweiter nachträg zur verbreitung der hypogaeen in russland. bull. soc. imp. nat. moscou 21 (4): 431–492. 138 e. kutorga and m. kataržytė castellano m. a., trappe j. m., luoma d. l. 2004. sequestrate fungi. (in:) g. m. mueller, g. f. bills, m. s. foster (eds). biodiversity of fungi. inventory and monitoring methods: 197-213. elsevier academic press, amsterdam. kataržytė m., kutorga e. 2005. three species of hypogeous fungi new to lithuania. botanica lithuanica 11 (4): 235–239. kirk p.m., cannon p.f., minter d.w., stalpers j.a. 2008. ainsworth & bisby’s dictionary of the fungi. 10th ed. cab international, wallingford, uk. kutorga e. 1994. new records in the genus geopora from lithuania. ekologija 2: 28-30. kutorga e. 2000. lietuvos grybai (mycota lithuaniae) 3 (5): ausūniečiai (pezizales). valstiečių laikraštis, vilnius. kutorga e. 2004. smiltyninis ausūnis naglių gamtos rezervate. raudoni lapai 8: 70. lange m. 1956. danish hypogeous macromycetes. dansk botanisk arkiv 16 (1): 5–84. ławrynowicz m. 1988. flora polska. grzyby (mycota) 18: workowce (ascomycetes), jeleniakowe (elaphomycetales), truflowe (tuberales). pwn, warszawa – kraków. ławrynowicz m. 1990. chorology of the european hypogeous ascomycetes. ii. tuberales. acta mycol. 26 (1): 7–75. martín m. p. 2001. chorologic database of european rhizopogon species. mycotaxon 78: 191–244. mazelaitis j. 1961. medžiaga lietuvos tsr gasteromicetų (gasteromycetes) florai. lietuvos tsr mokslų akademijos darbai. ser. c, 2 (25): 47–51. mazelaitis j. 1966. untersuchungen über der die schlauchpilzflora (ascomycetes) der litauischen ssr. (in:) gelehrte schriften 74, botanik. 2 auflage. referate des 3 myko-lichenologischen symposiums der baltischen sowjetrepubliken: 77-83. zvaigzne, riga. mazelaitis j. 1982. lietuvos tsr gasteromicetai. mokslas, vilnius. mazelaitis j., minkevičius a. 1957. valgomieji ir nuodingieji grybai. valstybinė politinės ir mokslinės literatūros leidykla, vilnius. mowszowicz j. 1957. conspectus florae vilnensis. przegląd flory wileńskiej. 1. wstęp i flora zarodnikowa okolic wilna. łódzkie towarzystwo naukowe, wydz. iii, 47:46–130, łódź. pegler d. n., spooner b. m., young t. w. k. 1993. british truffles. a revision of british hypogeous fungi. royal botanic gardens, kew. sosin p. e. 1973. opredelitel’ gasteromicetov sssr. nauka, leningrad. trzebiński j. 1934. spis wyższych grzybów podstawczaków i workowców, zebranych w wilnie i okolicach w latach 1925-32. prace tow. przyjaciół nauk w wilnie (wilno) 8 (4): 171–184. 2014-01-01t11:47:45+0100 polish botanical society the occurrence of cellulolytic fungi and fusarium in nests of circus pygargus teresa korniłłowicz-kowalska1, ignacy kitowski2 and helena iglik1 department of agricultural microbiology, mycological laboratory university of life sciences in lublin, leszczyńskiego 7 pl20-069 lublin, teresa.kornilowicz@up.lublin.pl 2 department of nature conservation, institute of biology university of maria curie-skłodowska, akademicka 19 pl20-033 lublin, ignacyk@autograf.pl korniłłowicz-kowalska t., kitowski i., iglik h.: the occurrence of cellulolytic fungi and fusarium in nests of circus pygargus. acta mycol. 45 (1): 97–114, 2010. a total of 45 species of cellulolytic fungi and ten fusarium species were identified in seven nests of montagu’s harrier. three genera (chaetomium, trichoderma, fusarium) represented 80% of the frequency of cellulolytic fungi. of them, chaetomium globosum, trichoderma viride and t. koningii were some of the most frequent species. a high differentiation of the richness and frequency of species of cellulolytic fungi depending on the nest and its individual layers was observed. reasons for the differences in the frequency and species composition of the fungi were discussed. key words: fungi, species richnes, cellulolytic, abilities, genus fusarium, nest material, wildliving birds introduction nests of the montagu’s harrier circus pygargus contain high amounts of plant material rich in cellulose. in favourable humidity conditions, a high cellulose content in the nest material provides a good substrate for the growth of cellulolytic fungi producing extracellular cellulolytic enzymes with endoglucanase and exoglucanase activity (cellobiohydrolase and β-glucosidase), hydrolysing cellulose to monosaccharides. these enzymes are produced by numerous fungi occupying the soil and plant remains occurring in the soil and its surface or colonising tissues of living plants. many of such fungi are represented by saprobes, e.g., chaetomium or trichoderma, while others, e.g., fusarium, are phytopathogens or potential phytopathogens (domsch et al. 1980; ghos, ghos 1992; korniłłowicz-kowalska et al. 2003). fungi of acta mycologica vol. 45 (1): 97–114 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 98 t. korniłłowicz-kowalska et al. the genus fusarium are causative agents of plant diseases such as wilt, seedling blight or head blight (vesonder, golinski 1989; kwaśna et al. 1991) and produce mycotoxins, e.g. trichotecenes, zearalenone or fumonisins, responsible for mycotoxicoses in humans and animals. the current knowledge of the occurrence of cellololytic fungi and fusarium representatives in birds’ nests is relatively poor (hubalek et al. 1973; hubalek 1974; hubalek, balat 1974). little is also known on the influence of physico-chemical properties such as ph and humidity on the frequency and species composition of these macromycetes in birds’ nests. studies have usually been conducted on nests of passeriformes (hubalek et al. 1973; hubalek 1974; hubalek, balat 1974) in which conditions differ from those in nests of water birds and wetland birds due to biotope types occupied by the birds. nests of montagu’s harrier in wetland habitats are therefore interesting objects of mycological studies. montagu’s harrier is a diurnal raptor which nests in different habitat types: steppes, open marshes as well as corn fields or young plantations of coniferous trees in europe and on other continents where it occurs (cramp, simmons 1980; clarke 1996). the bird populates entire poland, largely lowland regions avoiding distinctly mountainous ones. its present population size is estimated at 1 300-1 500 breeding pairs (tomiałojć, stawarczyk 2003). it occupies a broad spectrum of habitats in poland: wet open habitats such as marshes associated with wide river valleys, neglected fish ponds, willow (salix spp.) bushes in river valleys. the species nests in corn fields in many areas in poland; the trend has intensified in the last few years (tomiałojć, stawarczyk 2003). korniłłowicz-kowalska and kitowski (2009) investigated nests of montagu’s harrier in wetland habitats concentrating on the so called total frequency and species composition of saprotrophic fungi and fungi potentially pathogenic to humans and animals. it was shown that populations of ubiquistic species with a broad spectrum of substrates were some of the most frequent species, including thermotolerant species belonging to opportunistic pathogens, e.g., scopulariopsis brevicaulis and aspergillus fumigatus. the frequency of the latter, which also constitutes a high risk to birds, was high in some nests. among saptrotrophic fungi, populations of a few fungi known for cellulolytic abilities, mostly trichoderma spp., had a high occurrence frequency. the aim of this study was to examine the frequency and species composition of cellulolytic fungi and fusarium representatives in nests of montagu’s harrier with regard to the influence of some ecological factors. material and methods nests and nest material preparation. seven nests of montagu’s harrier from the błota serebryskie fens collected in 2004 (two nests: i and ii) and 2005 (five nests: iii,iv,v,vi and vii) were examined. the błota serebryskie fens are some of peat bogs of calcareous fens near chełm (torfowiska węglanowe koło chełma) (east poland: 51° 07’–51° 11’ n, 23° 30’–23° 42’ e). the occurrence of cellulolytic fungi 99 the błota serebryskie fens are lowland bogs lying on caco3 beds. bogs are dominated by the cladietum marisci community. water table levels range from 40 to 10 cm (spring) and from 20 to 0 (summer); water ph=7.7-8.6 (buczek 2005). the fens are protected by environmental law as an area important for bird occurrence. 14-42 pairs of montagu’s harrier nest here annually only in saw sedge (cladium mariscus) fields (krogulec 1994; kitowski 2002; kitowski unpublished data). at the time of laying, the nest is ovalor circle-like with a diameter of 20-56 cm (krogulec 1994; kitowski unpublished data). willow (salix sp.) or birch (betula sp.) twigs and dry stems of reed (phragmites australis) 10-15 cm in length as well as other plants, e.g., common broom (sarothamnus scoparius) and goosefoots (chenopodium sp.), are the base and the edge of the nest. its interior is lined almost exclusively with saw sedge leaves (cladium mariscus). however, the centre of the nest where the eggs are laid is lined with grass blades, rhizomes of agropyron sp. and rootlets. at the end of laying, the nest is a circle with a diameter of ca. 40 cm (22-51 cm) 10-15 cm of which is taken up by the lining (kitowski unpublished data). during breeding the nest lining is supplied by feathers from moulting, pellets and down shed by growing chicks, prey remains (hair, feathers, bones) of rodents (rodentia) and small birds as well as chitinous body parts of insects. entire nests or the material shaken out from one nest (nest vii) were collected after ca. 7-10 days after chick fledging when the nests no longer played important ecological and behavioural roles (kitowski 2002). nest material was sampled for mycological examinations from the lining (inner layer), nest edge (outer layer) and the intermediate layer between them (middle layer) (pugh 1966). samples were collected from a few places in each layer, a total of 100-200 g, were averaged and homogenised. the entire nest was shaken out from nest vii with a poorly developed lining and the material was used to prepare the initial sample similarly to individual layers. a total of 19 nest material samples were prepared. isolation and identification of fungi. cellulolytic fungi were isolated with the dilution method and by laying fine fragments (no longer than 0.5 cm in length) of the nest material on a cellulose substrate (whatman 1 filter paper) as the only source of c and energy containing (g × dcm-3) nh4no3 – 20, kno3 – 1.0, kh2po4 – 1.0, kcl – 0.5, mgso4 × 7h2o – 0.5, feso4 – traces, nacl – traces, agar – 20.0, streptomycin – 0.03%, chlortetracycline – 0.002%, ph – 5.5. forty fragments were analysed for each layer. fungi belonging to the genus fusarium were isolated with the dilution method on the nash and snyder medium (1952) containing (g × dcm-3) peptone – 15.0, kh2po4 – 1.0, mgso4 × 7h2o – 0.5, agar – 20.0, pcnb – 10.0, streptomycin – 0.03%. cellulolytic fungi and fusarium were cultured at 26°c. the frequency of fungi (nests collected in 2005) was determined as the mean values of cfu. g-1 of the dry weight of the nest material. the dry weight was determined with the weight method at 105°c. the rate of colonisation of the nest material by cellulolytic fungi was determined in %. growing pure fungal cultures were transferred onto pda slants (fusarium) or cellulose slants (without antibiotics). species identification of the isolates was based on macroand microscopic observations conducted on plates and in microcultures using pda, czapek-dox medium and selective nutrient agar (sna) for fusarium. a variety of systematic studies was used for 100 t. korniłłowicz-kowalska et al. the final classification of the fungi (domsch et al. 1980; nelson et al. 1983; kwaśna et al. 1991). determination of humidity and chemical properties of the nests. water content was determined with the weight method at 105°c. the following chemical analyses were performed: ph in h2o and kcl, organic c content (tiurin method), total c and total s content (elemental analysis by combustion analysis; a thermal conductivity detector), total n, total p, k, ca, mg after sample mineralization using the wet assay method in a mixture of concentrated h2so4 and perhydrol (n-total, p-total flow spectrophotometry; k, ca, mg atomic absorption spectroscopy method). the data are presented in table 1. the results giving the number of fungi belonging to the two groups (cellulolytic fungi and fusarium) were analysed by the statistical method counting standard deviations. the species diversity of cellulolytic fungi and fusarium based on the number of isolates of fungi representing individual species was analysed by calculating simpson’s index (krebs 1994) according to the formula: where pi is the share of isolates (strains) of species „i” in a fungal community (an entire nest or a layer of it) and is the quotient of the number of strains of the species and the number of isolates of all fungi obtained on an isolation medium (for cellulolytic fungi isolated with the dilution method and for fungi of the genus fusarium). values of simpson’s index range from 0 to 1-1/s, where s is the number of species in a community of fungi. species domination (trojan 1975) was determined using the formula: d = 100 . (sa : s), where s – number or isolates of species “a”, s – the sum of isolates of a group (cellulolytic fungi, fusarium). the frequency scale of species and taxonomic groups of fungi was as follows: sporadically < 1%, rarely 1-10%, frequently 11-25%, numerously 26-50%, very numerously > 50%. table 1 chemical content of montagu’s harrier (cyrcus pygargus) nests nest number content (% of dry weight) ph humidity (%)c total c org. n total s total p k ca mg h2o kcl iii 45.97 44.14 2.50 0.36 0.19 0.20 0.76 0.046 24.51 iv 26.86 25.74 1.31 0.16 0.08 0.22 1.95 0.144 49.64 v 44.42 41.60 1.52 0.19 0.14 0.19 1.08 0.059 7.43 (7.60; 7.36; 7.33) 7.46 (7.50; 7.47; 7.40) 70.65 vi 44.43 41.14 2.72 0.42 0.40 0.25 1.35 0.080 7.51 (7.26; 7.70; 7.58) 7.52 (7.35; 7.60; 7.60) 70.30 vii 6.70 7.10 52.50 the occurrence of cellulolytic fungi 101 results the frequency and colonisation rate of the nest material by cellulolytic fungi. the frequency of cellulolytic fungi (mean values) in the nests (i–iv) ranged between 220 and 300 000 cfu in 1 g of dry weight. over 1 mln cfu . g-1 of dry weight of the nest material was obtained for nest vii, where the material for analysis was shaken out. a higher frequency of fungi obtained for nest v results from the accumulation of fungal propagules following the shaking. the analysis of the frequency of fungi in individual layers of the nests showed that the lowest frequencies of cellulolytic fungi were recorded in the outer layer and the highest frequencies were recorded in the lining (internal layer) of the nests with the exception of nest iii (fig. 1a). the colonisation rate of the nest material by cellulolytic fungi was very high and ranged from 92.5% to 100% (fig. 2). the frequency of fusarium in the nests. the frequency of fungi isolated on the nash and snyder medium, preferential for the genus fusarium, ranged from 210 to 730 000 cfu . g-1 d.w. of the nest material, of which fusarium populations constituted between 10% (ni) and 50% (nii, niii) (fig.1b). other frequencies were consistent with those of other fungi growing on the nash and synder medium. the species composition of these fungi is given in tab. 7. the most numerous colonisation by fusarium representatives was observed in the linfig. 1. the preavalence rate of fungi in the nests of montagu’s harrier (cfu . 104 . g-1 d.w.): a – on the waksman’s medium for cellulolytic fungi, b – on the substrate for the genus fusarium. explanations: i1, ii1, iii1, iv1 – the outer layer of the nest; i2, ii2, iii2, iv2 – the intermediate layer of the nest; i3, ii3, iii3, iv3 – the inner layer of the nest (the core). 102 t. korniłłowicz-kowalska et al. ing and the outer layer while the weakest growth of fusarium was observed in the intermediate layer (fig. 1b). the species composition of cellulolytic fungi. cellulolytic fungi occupying the nests were represented by 45 species of 20 genera (tab. 2). a total of 1 276 pure cultures were differentiated; species of only five isolates of the genus penicillium were not identified. the diversity of cellulolytic species obtained with the dilution method (40 species) was greater than that with the fragment method (20 species) (tabs 3–4). cellulolytic fungi were isolated using the latter method from their metabolically active forms (hyphae) overgrowing the nest material. the most numerous colonisation of the nests was observed for cellulolytic fungi of the genera chaetomium, trichoderma and fusarium (tabs 2–3). their share in the isolates of cellulolytic fungi isolated with the dilution method was 44%, 20% and 19%, respectively, that is in total 83% of all cellulolytic fungi isolated with this method. the share of the genera was 44%, 11% and 32%, respectively, for nest material fragments laid on the medium for cellulolytic fungi. the species composition of the cellulolytic mycobiota differed depending on the nest. from 3 (v and vii) to 17 and 20 (ii and i) species were recorded in individual nests when the dilution method was used and from 3 to 14 species when the fragment method was used (tabs 3–4). differences in the species composition of cellulolytic fungi also depended on the study year. nests collected in 2004 were occupied by a community of cellulolytic fungi richer in species than those collected in 2005: 28 and 20 isolated species (dilution method), respectively – tables 3–4. the species richness of cellulolytic fungi in the nests, including frequencies of individual species, was analysed using simpson’s index (d) for four out of seven nests. the analysis indicated a considerable differentiation of species frequencies for both isolation techniques (tabs 3–4). the greatest differentiation of simpson’s indices was obtained for cellulolytic fungi isolated with the dilution method (0.289-nv to 0.758-niii), that is three and ten species (tabs 5–6). smaller differences in simpson’s indices were recorded for frequencies of species isolated with the fragment method: (0.578-nv to 0.857-nvii) with the number of species 6 and 14, respectively (tabs 5–7). despite a considerable dispersion of the values of simpson’s indices among individual nests and depending on the isolation method, mean values of the index for the nests total were similar: 0.876 (fragment method) and 0.866 (dilution method). this shows a high frequency richness of species of cellulolytic fungi occupying the nests. this was also observed fig. 2. the colonisation rate of the nest material by cellulolytic fungi (in %). the occurrence of cellulolytic fungi 103 in relation to individual layers of the nests. mean simpson’s indices for each layer (niv – nvi) were 0.798 – 0.865 for the fragment method and 0.786 – 0.877 for the dilution method (tab. 5). the analysis of the frequencies of individual species of cellulolytic fungi (tabs 3–4) showed that the greatest numbers of three chaetomium species: ch. table 2 a list of species of cellulolytic fungi isolated from nests of montagu’s harrier (circus pygargus) no. fungal species isolated with the dilution method isolated with the fragment method 1 acremonium strictum w. gams + 2 alternaria alternata (fr.) keissler + + 3 aspergillus fumigatus fres. + 4 botryotrichum piluliferum sacc.& march. + 5 chaetomium botrychodes zopf + + 6 ch. cochlioides pall. + + 7 ch. elatum kunze ex steud. + + 8 ch. funicola cooke + 9 ch. globosum kunze ex. steud. + + 10 ch. indicum corda + + 11 ch. piluliferum j. daniels + 12 chrysosporium pannorum (link) hughes + + 13 cladosporium cladosporioides (fres.) de vries + 14 cl. sphaerospermum penz. + 15 cylindrocarpon destructans (zinssm.) scholten + 16 doratomyces microsporus (sacc.) morton & g. sm. + 17 d. stemonites (pers. ex steud) morton & g. sm. + 18 fusarium avenaceum (fr.) sacc. + + 19 f. culmorum (w.g. smith) sacc. + + 20 f. graminearum schwabe + 21 f. (microdochium) nivale (fr.) ces. + 22 f. oxysporum schlecht. emend. sny & hans + + 23 f. poae (peck) wollenw. + 24 f. sacchari (butler.) w.gams + 25 f. sambucinum fuckel + 26 f. solani (mart.) appel et wollenw. emend sny et hans + + 27 f. sporotrichioides sherb. + 28 f. tricinctum (corda) sacc. + 29 gliocladium catenulatum gilm.&abbott + 30 g. roseum bain. + + 31 paeciliomyces carneus (duche&heim) a.h.s. brown& g.sm. + 32 penicillium brevicompactum dierckx + 33 p. chrysogenum thom + 34 p. expansum link ex gray + + 35 p. purpurogenum stoll + 36 penicillium sp. + 37 periconia atra corda + 38 pithomyces chartarum (berk.&curt.) m.b. ellis + + 39 phoma herbarum westend. + 40 scopulariopsis acremonium (delacr.) vuill. + 41 s. brevicaulis (sacc.) bain. + 42 thermomyces lanuginosus tsiklinsky + 43 thielavia heterotallica klopotek + 44 trichoderma koningii oudem + + 45 t. viride pers. ex gray + + 46 verticillium albo-atrum reinke&berthold + + 47 v. tenerum (nees ex pers.) link + 104 t. korniłłowicz-kowalska et al. ta bl e 3 t he c om po si ti on a nd fr eq ue nc y of s pe ci es o f c el lu lo ly ti c fu ng i i so la te d fr om n es ts o f m on ta gu ’s h ar ri er ( di lu ti on m et ho d) n o. fu ng al s pe ci es i ii ii i iv v v i v ii to ta l 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 a cr em on iu m s tr ic tu m 3* 1 2 6 2 a lte rn ar ia a lte rn at a 2 2 3 3 1 11 3 a sp er gi llu s fu m ig at us 3 3 4 b ot ry ot ri ch um p ilu lif er um 2 1 5 8 5 c ha et om iu m b os tr yc ho de s 1 1 6 c h. c oc hl io id es 10 10 14 34 7 c h. e la tu m 3 3 7 3 4 6 26 8 c h. fu ni co la 14 4 3 27 48 9 c h. g lo bo su m 2 9 3 3 7 4 18 31 7 27 16 18 14 5 10 c h. in di cu m 1 2 3 11 c h. p ilu lif er um 2 6 8 1 17 12 c hr ys os po ri um p an no ru m 5 2 7 13 c la do sp or iu m c la do sp or oi de s 3 8 11 14 c l. sp ha er os pe rm um 1 3 1 4 1 10 15 d or at om yc es s te m on ite s 1 3 10 4 8 1 4 31 16 fu sa ri um a ve na ce um 1 1 17 f. c ul m or um 2 2 18 f. g ra m in ea ru m 1 1 19 f. ( m ic ro do ch iu m ) ni va le 5 5 20 f. o xy sp or um 10 5 3 18 21 f. p oa e 2 2 5 4 13 22 f. s am bu ci nu m 2 3 1 6 23 f. s ol an i 2 1 6 9 18 24 f. s po ro tr ic hi oi de s 2 8 10 25 f. tr ic in ct um 2 13 9 25 49 26 g lio cl ad iu m c at en ul at um 1 1 27 g . r os eu m 1 2 3 28 pe ni ci lli um b re vi co m pa ct um 3 3 29 p. c hr ys og en um 1 1 30 p. e xp an su m 1 1 31 p. p ur pu ro ge nu m 1 1 32 p ho m a hi be rn ic a 10 10 33 sc op ul ar io ps is a cr em on iu m 1 1 34 s. b re vi ca ul is 2 2 35 t he rm om yc es la nu gi no su s 1 3 4 36 t hi el av ia h et er ot al lic a 1 1 the occurrence of cellulolytic fungi 105 37 tr ic ho de rm a ko ni ng ii 27 32 59 38 t. v ir id e 8 22 5 1 4 3 12 10 5 70 39 ve rt ic ill iu m a lb oat ru m 2 2 40 v. te ne ru m 1 1 8 10 to ta l 35 21 56 13 61 57 60 45 31 31 24 39 10 19 35 12 39 37 28 65 3 11 2 13 1 13 6 94 64 88 l eg en d: i ii , i v , v , v i, v ii – n es t n um be r; 1 .2 .3 – n es t l ay er ; 1 -o ut er . 2 -m id dl e. 3 -l in in g; * n um be r of is ol at es ta bl e 4 t he c ol on is at io n fr eq ue nc y of th e m at er ia l i n ne st s of m on ta gu h ar ri er b y in di vi du al s pe ci es o f c el lu lo ly ti c fu ng i ( fr ag m en t m et ho d) n o. fu ng al s pe ci es ii i iv v v i v ii to ta l 1 2 3 1 2 3 1 2 3 1 2 3 1 a lte rn ar ia a lte rn at a 5* 5 2 b ot ry ot ri ch um p ilu lif er um 2 6 3 11 3 c ha et om iu m c oc hl io id es 3 6 12 5 5 31 4 c h. e la tu m 3 3 5 c h. fu ni co la 3 7 12 4 2 28 6 c h. g lo bo su m 24 31 35 11 26 40 20 18 7 7 c h. p ilu lif er um 6 5 4 15 8 c yl in dr oc ar po n de st ru ct an s 1 1 9 fu sa ri um o xy sp or um 8 8 10 f. s ac ch ar i 2 1 5 5 13 11 f. s ol an i 3 7 5 5 20 12 f. s po ro tr ic hi oi de s 5 9 1 3 8 26 13 g lio cl ad iu m c at en ul at um ) 16 3 6 25 14 g . r os eu m 5 13 12 1 31 15 pa ec ili om yc es c ar ne us 7 1 8 16 pe ni ci lli um e xp an su m 1 1 17 pe ni ci lli um s p. 5 5 18 pe ri co ni a at ra 4 4 19 p ith om yc es c ha rt ar um 6 6 20 tr ic ho de rm a ko ni ng ii 44 24 11 13 12 2 10 6 21 t. v ir id e 2 12 9 8 15 11 2 11 11 14 95 22 ve rt ic ill iu m a lb oat ru m 3 3 to ta l 71 49 59 51 66 46 45 41 51 36 31 44 42 63 2 17 9 16 3 13 7 11 1 l eg en d as in t ab le 3 106 t. korniłłowicz-kowalska et al. globosum, ch. funicola and ch. cochlioides, as well as two trichoderma species: t. koningii and t. viride, colonised the nests. populations of fusarium: f. avenaceum, f. oxysporum, f. poae and f. tricinctum, gliocladium: g. roseum and g. catenulatum, as well as doratomyces stemonites were less numerous. the frequency distribution of individual cellulomycete species given above (as well as a few others) was uneven (tabs 3–4). the frequency of a species population was high in some nests while it was low in others or the species was altogether absent. chaetomium globosum was highly numerous in some nests and even its mass occurrence was observed. the frequency of this typical cellulolytic fungus was 66%, 69% and 48% of the total frequency of cellulolytic fungi when isolated with the dilution method in nests v, vi and vii, respectively, while its population constituted only 10% in nests i and ii and the fungus was not recorded in nests iii and iv. ch. cochlioides, ch. funicola and ch. elatum were more frequently recorded chaetomium representatives, although they were less numerous than ch. globosum (tabs 3–4). ch. cochlioides occupied four out of seven nests. the contribution of the species to the cellulomycete community ranged from 6% (niv) to 12% (niii) for the fragment method and from 10 to 16%, respectively, for the dilution method. ch. funicola was isolated mostly from nests collected in 2004 (ni, nii) and its frequency was ca. 20% of total cellulolytic fungi in these nests. c. piluliferum and its anamorph botryotrichum piluliferum which represented even 21% of the frequency of cellulolytic fungi in ni, and ch. elatum, which represented 5% and 12% of it in ni and nii, respectively, were also isolated from these nests. ch. indicum, ca. 2% of the population of cellulolytic fungi in ni, was one of the least frequently recorded representatives of chaetomium. the data (tabs 3–4) also show that the nests collected in 2004 (ni, nii) were characterised by a diversity of chaetomium species greater than that recorded in 2005 (niii-vii). of the two trichoderma species recorded in the nests, t. koningii had a slightly greater frequency than t. viride: 17%-44% and 10%-33%, respectively. t. koningii was, however, less widespread (two nests) than t. viride, which colonised six out of seven nests (tabs 3–4). ten fusarium species were recorded within cellulolytic fungi in the nests. they mostly occurred in nests iii and iv. fusarium tricinctum had the greatest frequency, however, only in nest iv. the cellulolytic population of this species represented 50% of total cellulolytic fungi in the nest. f. oxysporum was also recorded more frequently. table 5 simpson’s indices (d) for cellulolytic fungi in entire nests and individual layers of the nests of montagu’s harrier (circus pygargus) nest isolation method culture from dilutions fragment laying iii 0.758 0.761 iv 0.698 0.857 v 0.289 0.732 vi 0.590 0.578 iii-vi total 0.866 0.876 nest layer outer 0.877 0.865 middle 0.786 0.861 lining 0.831 0.798 the occurrence of cellulolytic fungi 107 ta bl e 6 t he c om po si ti on a nd fr eq ue nc y of f us ar iu m p op ul at io ns ( n as h an d sn yd er m ed iu m ) in n es ts o f m on ta gu ’s h ar ri er n o. fu ng al s pe ci es ii i iv v v i v ii to ta l 1 2 3 1 2 3 1 2 3 1 2 3 1 fu sa ri um a qu ed uc tu m 5 5 5 15 2 f. a ve na ce um 3 10 8 27 2 50 3 f. e qu is et i 7 7 4 f. o xy sp or um 22 6 11 1 40 5 f. p oa e 4* 4 8 3 13 3 8 10 53 6 f. s ac ch ar i 8 8 7 f. s em ite ct um 2 2 8 f. s ol an i 3 3 9 f. s po ro tr ic hi oi de s 4 1 2 8 15 10 f. tr ic in ct um 2 2 4 9 17 to ta l 6 4 9 43 23 51 13 5 11 16 14 5 10 21 0 19 11 7 29 35 l eg en d as in t ab le 3 ta bl e 7 t he fr eq ue nc y of o th er ( no nfu sa ri um ) fu ng al s pe ci es is ol at ed fr om n es ts o f m on ta gu ’s h ar ri er o n th e n as h an d sy nd er m ed iu m n o. fu ng al s pe ci es ii i iv v v i v ii 1 to ta l 1 2 3 1 1 2 3 1 1 2 3 1 a cr em on iu m k ili en se 2 14 13 17 5 42 12 10 5 2 a . s tr ic tu m 8 15 64 24 14 11 10 11 9 16 6 3 a cr em on iu m s p. 4 4 4 b ot ry tis c in er ea 5 5 5 m or tie re lla s p. 6 6 6 m uc or h ie m al is 3 3 7 r hi zo ct on ia s ol an i 23 12 35 8 tr ic ho de rm a ko ni ng ii 9 7 16 16 48 9 t. v ir id e 3 3 17 23 10 ve rt ic ill iu m a lb oat ru m 7 15 22 11 v. d ah lia e 12 12 12 v. le ca ni 4 4 13 n on s po ru la ti ng (d ar k m yc el iu m ) 22 14 10 6 19 4 3 6 16 23 12 3 to ta l 36 71 99 30 45 11 18 20 22 57 15 81 51 55 6 20 6 86 60 15 3 l eg en d as in t ab le 3 108 t. korniłłowicz-kowalska et al. its frequency was 16% of cellulomycetes in nest iv (tab. 3). a greater frequency of individual fusarium species was obtained when the selective isolation method for the genus was used (tab. 6). f. poae, f. avenaceum and f. oxysporum, 25%, 24% and 19% of the total fusarium frequency were the most frequently recorded species in the five nests collected in 2005. a smaller share of ca. 8% of fusarium populations on average was observed for f. aqueductum, f. sporotrichioides and f. tricinctum. low frequency of f. tricinctum isolation on the preferential medium for fusarium (nash and snyder medium with pcnb) in comparison with the medium used to isolate cellulolytic fungi indicates intrageneric competition in which f. tricinctum is a poorer competitor. on the other hand, this indicates a more frequent incidence of cellulolitic abilities within f. tricinctum than in other fusarium populations in the study environment (tabs 3–5). a frequency analysis of fusarium isolated on the nash and snyder medium confirmed that the greatest frequency and richness of fusarium species (six species) was observed in nest iv, similarly to the cellulose medium (tab. 3 and 5). the total frequency of these species in nest iv was as high as 56% of the total fusarium representatives in the nests (niv–vii). the lowest frequency (5%) and only one species, f. poae, was isolated on the nash and snyder medium from nest vii (the material was shaken out). fusarium poae was additionally the most widespread fusarium species in the nests isolated on the nash and snyder medium: it colonised four out of the five nests examined (tab. 6). both fusarium species and other species, loosely called accompanying species, were isolated on the nash and snyder medium: nine species altogether and two taxa identified only to the genus level. due to the fact that 14% isolates represented potentially phytopathogenic species in this group of fungi, their frequency was also analysed (tab. 7). these were in particular botrytis cinerea, rhizoctonia solani, verticillium albo-atrum, v. dahliae. acremonium, a genus related to fusarium, was isolated in this group. the fungus produces wettable 1-celled conidia, resembling microconidia in fusarium. the genus was very numerous and represented 36% of all isolates isolated on the nash and snydera medium (a total of 766) – table 7. discussion nutrient-specialised saprotrophic micromycetes, such as keratinolytic or cellulolytic fungi, in natural environments, e.g. in the soil, are distributed non-uniformly (korniłłowicz-kowalska, bohacz 2002a; korniłłowicz-kowalska et al. 2003). the uneven distribution of various physiological groups of fungal saprotrophs is conditioned by the dispersion of organic matter which is the source of food for the microorganisms. therefore, the role of examinations on the organic matter occurring in the environment is greater than that of examinations of the entire environment. nests of birds living in different biotopes are such microhabitats. they are usually agglomerations of different plant fragments (twigs, rootlets, grass blades, wooden pulp) and animal fragments (hair, feathers, bird faeces, chick down, prey remains, pellets). the nest mycobiota which comprises fungi that use simple organic compounds, e.g. representatives of mucorales and ubiquistic fungi (polyphages), is differentiated by the presence of compounds that use less accessible organic matter the occurrence of cellulolytic fungi 109 fractions such as ligninocellulose or keratine. both typically saprotrophic species and potentially pathogenic species, opportunistic pathogens causing diseases in humans and animals as well as toxicogenic fungi are found in fungal communities in birds’ nests (hubalek 1974; hubalek, balat 1974; pinowski et al. 1999; korniłłowiczkowalska, kitowski 2009). as observed in a previous study by the authors (korniłłowicz-kowalska, kitowski 2009), a high total frequency of saprotrophic micromycetes, a high richness and frequency of species with a broad spectrum of substrates (polyphages), sugar-loving fungi and species belonging to opportunistic pathogens in humans and animals were recorded in the nests of montagu’s harrier. the present study shows that nests of montagu’s harrier are characterised by a high frequency of fungi specialised in cellulose biodegradation. the number of propagule units of these fungi corresponded to the frequencies of cellulolytic fungi in composts consisting of plant material and hen feathers (korniłłowicz-kowalska, bohacz 2002b). they exceeded, however, the frequencies of these microorganisms in soils fertilised with natural fertiliser (manure) and were considered to be rich in these micro-organisms (korniłłowicz 1989). the high rate of colonisation of the nest material by cellulolytic fungi recorded here also indicates a high enzymatic activity of these micro-organisms and intensive processes of degradation of cellulose and other polysaccharides occurring in nests abandoned after breeding. identification examinations and lists of total simpson’s coefficients show a great differentiation of the composition and frequency of cellulolytic species of fungi, both in relation to entire nests and their individual layers. the high values of simpson’s coefficient show that the communities of cellulolytic fungi in this microhabitat are dominated by a relatively small number of populations of these fungi. this was also observed for the total mycobiota in nests of montagu’s harrier (korniłłowicz-kowalska, kitowski 2009). chaetomium spp. (ascomycetes), trichoderma spp. and fusarium spp. (mitosporic fungi) were the most numerous populations of cellulose-decomposing fungi. they constituted over 80% of total cellolytic fungi. representatives of these genera were often recorded among fungal colonisers of wild-living birds’ nests and plumage (pugh 1965; apinis, pugh 1966; mishra, tiwari 1970; hubalek 1974; hubalek et al. 1973; takatori, hasegawa 1981; kaul, sumbali 1999). however, those were mostly species belonging to passeriformes. chaetomium, representing so called soft rot fungi causing the destruction of the surface wood layer resulting form cellulose biodegradation, other polysaccharides and lignin structure injuries, is one of the most effective cellulose destruent (domsch et al. 1980). of the seven chaetomium species isolated from the nests, chaetomium globosum had the highest frequency and constituted 70-80% of the total cellulolytic micromycetes in some nests. populations of ch. cochlioides, ch. funicola and ch. elatum were less numerous (10-20%) in individual nests. the four chaetomium species are thought to be widespread, both in nests and on feathers of different bird species, mostly passeriformes (hubalek 1974). the analysis shows that the genus trichoderma was represented in the nests by t. viride and t. koningii, which reach 33% and 44% of total cellulolytic fungi in some nests. the fungi were previously infrequently isolated from birds’ nests. hubalek (1974) recorded t. viride in nests of song thrush (turdus philomelos) containing the lining of wooden pulp (lignocellulose – author’s note). t. viride was more often isolated from the plumage of passeriformes, also including turdus philomelos (pugh 110 t. korniłłowicz-kowalska et al. 1965; mishra, tiwari 1970). trichoderma koningii was recorded in nests of passer domesticus and a few other passeriformes birds (apinis, pugh 1967). fungi belonging to the genus fusarium were recorded by pugh (1986) as well as by takatori and hasegawa (1971) in nests and plumage of different species of terrestrial birds: passeriformes and columbiformes. according to hubalek (1974), however, fusarium mostly colonises nests of water birds. the genus was also recorded on feathers of water birds (pugh 1966). of fusarium species recorded in birds’ nests, only f. oxysporum, f. sporotrichioides and f. moniliforme have been listed (hubalek 1974). present examinations show that ten fusarium species with cellulolytic abilities occurred in the nests with different frequencies. examinations of the composition and frequency of fusarium species on the selective medium for the genus (nash and snyder medium) show that almost 70% of the fungi were constituted by populations of f. poae, f. avenaceum and f. oxysporum. cellulolytic gliocladium spp., mostly g. roseum, and darkly pigmenting fungi cladosporium herbarum, cl. cladosporoides, cl. sphaerospermum, doratomyces stemonites, d. microsporus, alternaria alternata, occurred less frequently in the nests (less than 10% of total records). the species were previously isolated from nests and (or) plumage of many songbirds (passeriformes) and water birds (pugh 1965; 1966; hubalek et al 1973; hubalek 1974; hubalek, balat 1974; takatori, hasegawa 1981). among them, doratomyces (stysanus) stemonites is believed to be closely related to water birds (nest and plumage) (hubalek 1974). cellulolytic fungi such as penicillium spp., aspergillus, including a. fumigatus, scopulariopsis spp. excluding s. acremonium and s. brevicaulis, and a few others were recorded sporadically in the nests. as a previous study by korniłłowicz-kowalska, kitowski (2009) shows, a. fumigatus and s. brevicaulis were recorded as frequent within so-called total fungi (martin’s medium) and potentially zoopathogenic fungi (sabouraud medium). in the light of the new data, a very low number of records of cellulolytic isolates of s. brevicaulis and especially a. fumigatus, which causes opportunistic infections of raptors (joseph 2000), shows the absence of a correlation between these fungi and cellulose substrates in the nests. this observation supports the suggestion proposed in a previous study (korniłłowicz-kowalska, kitowski 2009) that a. fumigatus is nutritionally related to keratin waste found in nests. of the total 20 genera and 45 species of fungi isolated, the majority are cosmopolitan species, widespread in soil, colonising the phyllosphere, dead plant remains or, like some, occurring in the air. they are mostly saprotrophs, e.g., chaetomium spp., trichoderma spp., gliocladium spp., doratomyces spp., penicillium spp., or potential phytopathogens: fusarium spp., verticillium spp., alternaria spp., cladosporium spp., rhizoctonia spp. (domsch et al. 1980). as noted above, many of these genera were isolated from the plumage and pellets, faeces and other animal remains (hubalek 1974; domsch et al. 1980). doratomyces stemonites, isolated with a very high frequency (over 50%) from pellets of montagu’s harrier in one of the nests (korniłłowicz-kowalska, kitowski 2009) as well as fusarium tricinctum, less widespread in the natural environment in comparison with other fusarium species, are noteworthy (domsch et al. 1980). of rarely recorded species, darkly pigmenting micromycetes such as periconia atra and pithomyces chartarum should be mentioned. the former was isolated together with many other species (e.g., fusarium oxysporum, f. sacchari, cladosporium spp., alternaria alternata, trichoderma viride) by mazurkiewicz-zapałowicz, wróbel and buczek the occurrence of cellulolytic fungi 111 (2008) from the saw sedge phyllosphere. as saw sedge leaves are used as the lining in nests of montagu’s harrier, we think that this plant material was a source of a considerable amount of cellulolytic fungi and fusarium. the results allow us to claim that humidity and thermal conditions recorded in the nests during the incubation and chick rearing led to the selection of certain populations of cellulolytic micromycetes. as table 1 shows, despite differences, nest humidity was high (23%–71%) and the ph ranged from neutral to slightly alkaline (phkcl 7.10–7.60). as indicated by the frequency of their occurrence, mostly fungi with a high water activity coefficient (wa) as well as thermotolerant and alkalotolerant fungi had favourable conditions in the microhabitat. similar observations were made in relation to nutritionally unspecialised fungi colonising nests of montagu’s harrier (korniłłowicz-kowalska, kitowski 2009). it was also observed that the frequency of cellulolytic fungi was greater in nests with a higher (nv–nvii) rather than a lower (niii–niv) humidity. this was caused by a frequency increase in chaetomium, trichoderma and fusarium populations considered to be so called tertiary colonisers which require at least wa = 0.9–0.95 for growth (grant et al. 1989). chaetomium globusom and trichoderma viride were especially hydrophilous (apinis, pugh 1966; hubalek 1974; domsch et al. 1980). the species did not occur in nest iii (with the lowest humidity, 23%–25%). doratomyces stemonites or trichoderma koningii, which had lower water condition requirements, had good growth conditions in this environment (domsch et al. 1980). t. koningii as well as ch. globosum, ch. cochlioides and ch. funicola are also thermotolerant (apinis, pugh 1966) and grow at 37°–38°c or even higher temperatures. it may be supposed that the selection of these fungi occurred as early as during incubation when the nest temperature can reach even 40–41°c (pinowski et al. 1999). the high nest humidity and temperature did not encourage the development of species such as penicillium which prefer environments with lower wa values (griffin 1972, 1994). the majority of species of this genus are also psychrophiles or psychrotrophs and acidophilous fungi (apinis, pugh 1966; griffin 1972; 1993). cladosporium spp. and alternaria spp. are also considered to be relatively psychrophilous or psychrotolerant (apinis, pugh 1966), which explains their low frequency in the nests within a short period after they were abandoned by the birds. the examinations show that over 40% of the species composition, that is 20 fungal species, of the nests are potential phytopathogens, mostly fusarium spp. species such as verticillium albo-atrum, v. dahliae, rhizoctonia solani, botrytis cinerea, alternaria alternata, cladosporium spp. were also recorded. a high contribution of potentially phytopathogenic fungi in the mycobiota of the nests was conditioned by the plant origin of the nest material, rich in cellulose and other polysaccharides. the development of these fungi as well as exclusively saprotrophic fungi was also encouraged by a high nitrogen and phosphorus content (tab.1) from bird faeces, feathers and other animal substrates. previous studies by korniłłowicz (1989); korniłłowiczkowalska et al. (2003) showed that the development of fusarium and cellulolytic fungi is stimulated by the presence of nitrogen and phosphorus in the soil. the present findings allow us to think that birds’ nests, including nests of montagu’s harrier, may be some of the links in the circulation of phytopathogenic fungi, such as fusarium spp., verticillium albo-atrum or v. dahliae. these fungi may survive, reproduce and, consequently, spread aerogenically, also by birds. in montagu’s 112 t. korniłłowicz-kowalska et al. harrier, the process is undoubtedly encouraged by high breeding densities that reach even 10.6 nest/100 hectares (krogulec 1992). conclusions nests of montagu’s harrier are characterised by a high frequency, colonisation • rate and species richness of cellulolytic fungi and fusarium. the number and frequency of fungal species differed depending on the nest and • the layer (outer, middle, lining). the highest frequency was recorded for fungal species with high water condition • requirements, often thermotolerant, of the genera: chaetomium, trichoderma, fusarium. species with lower water requirements, psychrophilous or psychrotolerant spe-• cies, such as penicillium spp., cladosporium spp., were recorded rarely and their frequency was lower. as observed, a neutral or slightly alkaline reaction of the nests encourages growth • of alkalotolerant species. acknowledgement. this work was supported by the by the committee of scientific researches, a grant no. 2p04g03330. references apinis a.e., pugh g.j.f. 1967. thermophilous fungi of birds’ nests. mycopathologia 33: 1–9. buczek t. 2004. błotniak łąkowy circus pygargus (in:) m. gramadzki (ed.).poradnik ochrony siedlisk i gatunków. natura 2000 – podręcznik metodyczny. ministerstwo środowiska, warszawa: 226–230. clarke r. 1996. montagu’s harrier. arlequin. chelmsford. cramp, simmons k.e.l. 1980. the birds of the western paleartic. volume 2. oxford university press, oxford, uk. domsch k.h., gams w., anderson t.h. 1980. compendium of soil fungi. i acad. press. london. ellis h.b. dematiaceaus hyphomycetes. 1971. commonwealth, mycological institute kew surrey, england. ghosh b., ghosh a. 1992. degradation of cellulose by fungal cellulase. (in:) g. winkelmann (ed.). microbial degradation of natural products vch weinheim 83–125. grant c., hunter c.a., flannigan b., bravery a.f. 1989. the moisture requirements of moulds isolated from domestic dwelling. int. biodet. 25: 259–284. griffin d.m. 1972. ecology of soil fungi. chapman and hall. england. griffin d.m. 1994. fungal physiology. i. willey – liss. new york usa. hubalek z. 1974. fungi associated with free–living birds in czechoslovakia and yugoslavia. acta sc. nat. brno 8 (3): 1–62. hubalek z., balat f. 1974. the survival of microfungi in the nests of tree sparrow (passer montagus l.) in the nest-boxes over the winter season. mycopathologia 54: 517–530. hubalek z., balat f., touškova i., vlk t. 1973. mycoflora of birds nests in nest-boxes. mycopath. mycol. appl. 49: 1–12. joseph v. 2000. aspergillosis in raptors. seminars in avian and exotic. pet medicine 9: 66–74. kaul s., sumbali g. 1999. impact of some ecological factors on the occurrence of poultry soil-inhabiting keratinophiles. mycopathologia 143: 155–159. kitowski i. 2002. behaviour of montagu’s harrier juveniles circus pygargus during the post-fledging dependency period in southeast poland. berkut 11: 201–207. the occurrence of cellulolytic fungi 113 korniłłowicz-kowalska t. 1989. wpływ intensywnego nawożenia obornikiem and granulatem keratynokoro-mocznikowym na wybrane zespoły mikroflory glebowej. zesz. probl. post. nauk rol. 370: 85–96. korniłłowicz-kowalska t., bohacz j. 2002a. some correlations between the occurrence frequency of keratinophilic fungi and selected soil properties. acta mycol. 37 (1/2): 101–116. korniłłowicz-kowalska t., bohacz j. 2002b. próba kompostowania odpadów pierza z zastosowaniem inokulum grzybowego. ii. dynamika rozwoju drobnoustrojów. acta agrophys. 73: 189–197. korniłłowicz-kowalska t., iglik h., wojdyło b. 2003. correlation between the abundance of cellulolitic fungi and selected soil properties. acta mycol. 38: 161–172. korniłłowicz-kowalska t., kitowski i. 2009. diversity of fungi in nests and pellets of montagu’s harrier (circus pygargus) from east poland – importance of hemical and ecological factors. ecol. chem. enngineer. s. (in press) krebs c.j. 1994. ecology. the experimental analysis of distribution and abundance, fourth edition. harper collins, new york, usa krogulec j. 1994. breeding ecology of montagu’s harrier circus pygargus on calcareous marshes near chelm. unpublished ph.d. thesis. university of maria curie sklodowska. lublin. (in polish). kwaśna h., chełkowski i., zajkowski p. 1991. flora polska. grzyby (mycota) 22: (deuteromycetes), (hyphomycetales), fusarium. pwn. warszawa. mazurkiewicz-zapałowicz k., buczek t. 2008. grzyby mikroskopowe związane z kłocią wiechowatą (cladium mariscus l.) pohl. chrońmy przyrodę ojczystą 64 (1): 45–57. mishra r.r., tewari r.p. 1970. fungal species associated with certain common birds. sci. cult. 36: 350–352. nash s.m., snyder w.c. 1962. quantitative estimation by plate counts of propagules of the bean root rot fusarium in field soils. phytophology 52: 567–572. nelson p.e., toussoun t.a., morasas w.f.o. 1983. fusarium species: an illustrated manual for identification. the pensylvania state, university press. pinowski j., pinowska b., haman a. 1999. fungi in birds’ plumage and nets. intern. stud. sparrows 26: 3–28. pugh g.j.e. 1965. cellulolytic and keratinophilic fungi recorded on birds. sabouraudia 4: 85–91. pugh g.j.e. 1966. associations between birds, nests, their ph and keratinophilic fungi. sabouraudia 5: 43–53. takatori k., hasegawa a. 1981. isolation of keratinophilic and non-keratinophilic fungi from birds nests. trans. mycol. soc. (japan) 22: 347–352. tomiałojć l., stawarczyk t. 2003. the avifauna of poland. distribution, numbers and trends. wroclaw, ptpp “pro natura” (in polish). trojan p. 1975. general ecology. wyd. pwn, warsaw (in polish). vesonder r.f., golinski p. 1989. metabolites of fusarium (in:) j. chełkowski (ed.). fusarium, mycotoxins, taxonomy and pathogenicity. elsevier, amsterdam: 1–40. występowanie grzybów celulolitycznych oraz fusarium w gniazdach circus pygargus streszczenie celem przedstawionej pracy było zbadanie liczebności oraz składu gatunkowego grzybów celulolitycznych i fusarium w gniazdach błotniaka łąkowego (circus pygargus, falconiformes). stosując metodę rozcieńczeń oraz wykładania fragmentów materiału gniazdowego na podłoża selektywne dla tych grzybów, zbadano 19 próbek z 7 gniazd zlokalizowanych na obszarze torfowisk węglanowych koło chełma w płd.-wsch. polsce (rezerwat błota serebryskie). próbki do badań pobierano z 3 warstw gniazda: wyściółki, warstwy zewnętrznej oraz środkowej leżącej pomiędzy nimi. 114 t. korniłłowicz-kowalska et al. stwierdzono, że gniazda błotniaka łąkowego cechuje wysoka ogólna liczebność grzybów celulolitycznych i fusarium, duże bogactwo oraz zróżnicowanie frekwencji ich gatunków. wykazano, że 80% zbiorowiska grzybów celulolitycznych reprezentowały 3 rodzaje: chaetomium, trichoderma i fusarium. spośród 45 zanotowanych gatunków najwyższą liczebnością wyróżniały się: chaetomium globosum, trichoderma koningii i t. viride. w obrębie fusarium najczęściej notowano: f. poae, f. avenaceum i f. oxysporum. odnotowane dominanty gatunkowe należały do grzybów hydrofilnych, na ogół termotolerancyjnych. niską frekwencją odznaczały się natomiast gatunki preferujące środowisko o niskiej aktywności wodnej, psychrofilne i acidofilne takie jak: cladosporium spp. i penicillium spp. 2014-01-01t11:50:39+0100 polish botanical society a confusing duo: calocybe cerina and callistosporium pinicola (agaricales) eef arnolds holthe 21, nl 9411 tn beilen, eefarnolds@hetnet.nl a r n o l d s e.: a confusing duo: calocybe cerina and callistosporium pinicola (agaricales). acta mycol. 41 (1): 29 40, 2006. the name calocybe cerina has recently been applied for two different species of agarics. after studying collections from different european countries it is concluded that the true calocybe cerina is very close to c. chrysenteron and best regarded as a variety. the new combination calocybe chrysenteron var. cerina is proposed and a full description of that taxon is given. the majority of collections, identified as c. cerina, appear to belong to a fungus, provisionally described by bon as callistosporium luteoolivaceum fo. minor. it is described here as a new species: callistosporium pinicola. calocybe juncicola is another species that might be confused with the two species mentioned before and therefore a full description is provided. the status of the genera rugosomyces and calocybe is discussed. the new combinations calocybe obscurata and calocybe pudica are made. key words: callistosporium, calocybe, rugosomyces introduction in the netherlands in recent years several collections were made of a fairly small, white-spored agaric with an orange-brown pileus and crowded, orange-yellow lamellae, growing on stumps of coniferous trees. it was initially identified as calocybe cerina (pers.: fr.) donk (a r n o l d s , b e c k e r 1993). an important indication was the presence of granules in the basidia that were apparently siderophilous, staining dark violet in acetocarmine with addition of iron. however, some characters were deviating from most descriptions in literature, in particular the striking red to violet staining of all tissues in koh and ammonia, the complete absence of clamp-connections and the lignicolous habitat, suggesting that it could be an undescribed taxon. recently i came across descriptions of callistosporium luteoolivaceum var. minor m. bon ined. (l u d w i g 2001; w i l h e l m 2003) that showed striking resemblance with our material, e.g. the red staining in ammonia and koh. in this paper i shall try to unravel the identity of the collected material and i will discuss the taxonomic position of calocybe cerina (pers.: fr.) donk and the related c. juncicola (heim) sing. acta mycologica vol. 41 (1): 29-40 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 30 e. arnolds what is calocybe cerina? agaricus cerinus was initially described by p e r s o o n (1801: 321) as a fairly small agaric, growing in pine forests (substrate not explicitly mentioned) with dark waxyellow (‘flavo-cerinus opacus’), fleshy, flattened to depressed pileus; crowded, narrow lamellae and a taste becoming bitter after a while. in his sanctioning description f r i e s (1821:89) mainly copied persoon’s description, adding that the context and spore print are white. most authors have interpreted the name as a species close to or identical with calocybe chrysenteron (bull.: fr.) sing. p e r s o o n (1801) treated agaricus chrysenterus bull. immediately after his a. cerinus. the two taxa are microscopically characterized by e.g very small, ellipsoid spores, a pileipellis in the form of a cutis with transitions to a trichodermium, and presence of clamp-connections. a different interpretation of agaricus cerinus was published by a r n o l d s & b e c k e r (1993) and accepted by e.g. k a l a m e e s (1995, 2004). the basidiocarps of that fungus occur on dead wood, are slightly smaller and thin-fleshed, their tissues turn reddish in alkaline solutions and clamp-connections are absent. after comparing collections and modern descriptions of these taxa i believe that the first-mentioned interpretation is in better agreement with the original diagnosis, mainly in view of the larger size of the basidiocarps, the thicker context and the substrate on soil and litter. therefore i present first a description of this little-known fungus. calocybe chrysenteron (bull.:fr.) sing. var. cerina (pers.: fr.) arnolds nov. comb. – fig. 1. basionym: agaricus cerinus pers.: fr., syst. mycol. 1: 89. 1821. agaricus cerinus pers., syn. meth. fung.: 321. 1801; agaricus cerinus pers.: fr., syst. mycol. 1: 89. 1821; tricholoma cerinum (pers.:fr.) quél., champ. jura vosges 1; 81. 187; tricholoma chrysenteron subsp. cerinum (pers.: fr.) konr. & m., icon. sel. fung. 3: pl. 267, fig. 2. 1937; calocybe cerina (pers.: fr.) donk in beih. nova hedwigia 5: 43. 1962; rugosomyces cerinus m. bon in doc. mycol. 21 (82): 66. 1991. – tricholoma pseudoflammula j. lange in dansk bot. ark. 8(3): 24. 1933; spec. pl.: 1173. 1753; calocybe pseudoflammula (j. lange) sing. in sydowia 15: 47. (‘1961’) 1962; rugosomyces pseudoflammula (j. lange) m. bon in doc. mycol. 21(82): 65. 1991. excl. – calocybe cerina sensu arnolds & becker in coolia 36: 70-73, fig. 1. 1993 ( callistosporium pinicola); sensu dermek in fung. rar. ic. col. 17: 10. 1987 ( c. fallax); sensu kalamees in scripta mycol. 18: 84. 2004 ( callistosporium pinicola); rugosomyces pseudoflammula sensu m. bon, fl. mycol. eur. 5, collybio-marasmïoïdes: 110. 1999 ( c. chrysenteron). sel. icon. – cetto, funghi vero 3: pl. 1024. 1979; flora batava 25 : pl. 1994a. 1920; konr. & m., icon. sel. fung. 3: pl. 267, fig. 2. 1927; j. lange, fl. agar. dan. 1: pl. 24b. 1935 (as t. pseudoflammula); schweizer pilzt. 5: pl. 42. 1972. sel. descr. & figs. – m. bon, fl. mycol. eur. 5, collybio-marasmïoïdes: 111. 1999; konr. & m., icon. sel. fung. 3: pl. 267 ii. 1927; j. lange, fl. agar. dan. 1: 57. 1935 (as t. pseudoflammula). pileus 12-40(-50) mm, convex at first, with thin, involute margin, then planoconvex to flattened or slightly depressed with straight, often undulating margin, with or without weak umbo, not hygrophanous, brownish yellow, brownish orange to orange-brown from the beginning (e.g. k. & w. 5b7, 5c8, 6c7, 6c8), dull and dry, a confusing duo: calocybe cerina and callistosporium pinicola (agaricales) 31 smooth to slightly tomentose, especially at centre, not striate. lamellae, l 38-60, l 1-7, adnexed, emarginate to adnate, very crowded, segmentiform, up to 5 mm broad, thin, lemonto golden-yellow or orangeish yellow, with concolorous, entire edge. stipe 20-50 x 2-6(-10) mm, cylindrical or compressed, often tapering to base, stuffed to fistulose, concolorous with pileus, sometimes weakly yellow striate lengthwise, at apex pruinose, at base white tomentose or strigose-hairy. context whitish, pale yellow to lemon-yellow. smell farinaceous when cut; taste first farinaceous, then often more or less bitter. spore print white. spores 2.5-4.0(-4.5) x 2.0-3.0 μm, av. 3.1-3.5 x 2.3-2.5 μm, q 1.2-1.5, qav. 1.31.4, broadly ellipsoid to ellipsoid. basidia 17-23 x 4.0-7.0 μm, clavate, 4-spored, with numerous small granules. lamella edge fertile. hymenophoral trama subregular, made up of hyphae with cylindrical or slightly inflated elements, 3.0-12 μm broad. pileipellis a cutis of repent hyphae, 2.0-6.0 μm broad, often with some broader, ellipsoid to subglobose cells, often at centre with trichodermial tufts of erect and ascending hyphae, with yellowish intracellular pigment. stipitipellis a cutis, made up of repent hyphae, 2.0-4.0 μm wide, at stipe apex with clusters of undifferentiated, erect hyphal tips up to 40 μm long, 2.0-5.0 μm wide. clamp-connections present. chemical reactions: no part of basidiocarp reddening with koh or ammonia. granules in basidia staining dark purple in acetocarmine with iron (siderophilous). habitat and distribution: mainly on needle litter in coniferous forests (picea, abies), also reported under broad-leaved-trees (quercus); on mesic to dry, calcareous soil. probably widespread in europe but much rarer than var. chrysenteron. exact distribution unknown in view of taxonomic confusion (see notes). with certainty recorded from denmark, germany, switzerland (k o n r a d & m a u b l a n c 1937) and the netherlands (flora batava, 1920). collections examined. – denmark: rubjerg, knude plantage, 26 sept. 1990, j. vesterholt 90-505 (c); same loc., 21 oct. 1991, j. vesterholt 91-801 (c) – germany: bavaria, frankische schweiz, ebermannstadt, aufsesstal, 19 sept. 2005, e. arnolds 05-87 (l). fig. 1. calocybe chrysenteron var. cerina. a. basidiocarps x 1; b. spores x 2000; c. basidia x 1000; d. pileipellis x 1000 (all from arnolds 05 87). 32 e. arnolds the studied collections differ from typical calocybe chrysenteron morphologically only in the duller, more brownish colours of pileus and stipe and the paler context. in var. chrysenteron all parts of the basidiocarps are bright yellow to orange-yellow (e.g. l u d w i g 2000). in addition var. cerina seems to be almost confined to coniferous forests, whereas var. chrysenteron is mostly (but not always) found in deciduous forests. both taxa are calciphilous. some modern authors synonymize c. chrysenteron and c. cerina (e.g. l u d w i g 2001), others treat them as separate species (e.g. b o n 1999; h o r a k 2005). the differences are small but seem to be rather constant and warrant in my opinion a distinction in the rank of variety. many modern authors regard tricholoma pseudoflammula j. lange as a synonym of calocybe chrysenteron (e.g. l u d w i g 2001; k a l a m e e s 2004), but lange’s plate (24b) is typical of var. cerina in view of the orange-brown pileus, already in young basidiocarps, and the pale context. for differences with c. cerina sensu arnolds and becker ( callistosporium pinicola) see notes on that species. a double of calocybe cerina: callistosporium pinicola callistosporium pinicola arnolds nov. spec. – fig. 2. pileus (5-)10-30 mm convexus margine involutus, dein planiusculus vel depressus, haud vel leviter hygrophanus, flavo-brunneus, aurantio-brunneus, rufo-brunneus, numquam striatus, siccus, centro tomentosus. lamellae adnatae vel emarginatae, confertae, tenuae, flavae vel aurantio-flavae. stipes (12-)14-32 x (1,5-)2-3,5 mm, cylindricus vel compressus, pileo subconcolorus. caro tenuis, fragilis, pallide flava. odor nullus vel farinaceus. sapor nullus vel amarescens. fig. 2. callistosporium pinicola. a, e. basidiocarps x 1; b, f. spores x 2000; c, g. basidia x 1000; d, h. pileipellis x 1000 (a d from arnolds 6100 (holotype), e h form a. becker, 6 oct. 1990). a confusing duo: calocybe cerina and callistosporium pinicola (agaricales) 33 sporae (2,5-)2,8-4,7 x 2,0-3,5 μm, ellipsoideae vel late ellipsoideae, laeves, hyalinae, saepe corpusculis xanthis vel brunneis. basidia 12-25 x 3,5-6,0 μm, clavatae, tetraspora. lamellarum trama regularis. cystidia nulla. pileipellis cutis vel trichodermium, hyphis 2,0-5,5 μm latis, interdum ex cellulis inflatae ad 8,0 μm. fibulae absentes. basidia, trama et pileipellis granulis numerosis brunneis in aqua, rubescentis vel violascentis in koh et ammonia. habitatio ad truncos putridos coniferarum. aestateautomno. holotypus: ‘the netherlands, dwingeloo, lheederzand, 4 oct. 1990, e. arnolds 6100’ (l). synon.: callistosporium xanthophyllum (malenc. & bert) m. bon fo. minor m. bon ad int., in flore mycol. europe 2: 83. 1991 (invalid, without latin diagnosis and without designation of type). misapplied names: calocybe cerina sensu arnolds & becker in coolia 36: 70. 1993; sensu kalamees in scripta mycol. 18: 84. 2004. sel. plates. – ludwig, pilzkompendium 1: pl. 15, fig. 6.2 a, c. 2000 (as callistosporium luteoolivaceum var. minor m. bon ined.); wilhelm in schweiz. z. pilzk. 81: 64. 2003 (as callistosporium luteoolivaceum var. minor m. bon). sel. descr. & figs. – arnolds & becker in coolia 36: 70-73, fig. 1. 1993 (as calocybe cerina); ludwig, pilzkompendium 1: 37-38. 2001 (as callistosporium luteoolivaceum var. minor m. bon ined.); wilhelm in schweiz. z. pilzk. 81: 62-66. 2003 (as callistosporium luteoolivaceum var. minor m. bon) . pileus (5-)10-30 mm, first convex with narrow, involute margin, then plano-convex to flattened with straight margin, often with slightly depressed centre, not or weakly hygrophanous, usually vividly yellow-brown, orange-brown, rusty-brown to red-brown, occasionally with olivaceous hue near centre (e.g. k. & w. 5d7, 6d7, 7e7; mu. 5 yr 6/6, 10 yr 4/6, 5/8), rarely entirely butter-yellow (k. & w. 4a5), not striate, dry, smooth or slightly radially fibrillose, often at centre tomentose. lamellae, l 28-36, l (1-)3-7, adnate to strongly emarginate, crowded to very crowded, segmentiform, up to 4 mm broad, thin, butter-yellow, golden-yellow to yellowish orange (mu. 2.5 y 6/8; k. & w. 4a5, 5c6, 4c6), with concolorous, entire edge, becoming dark red-brown to almost black in exsiccata. stipe (12-)14-32 x (1.5-)2-3.5 mm, cylindrical or sometimes compressed, solid or narrowly fistulose, concolorous with pileus or paler yellow-brown, slightly aerenchymatic at first, then smooth or white striate lengthwise, at apex slightly pruinose, at base often white tomentose. context in pileus thin, up to 2 mm thick, fragile, concolorous with surface, inside stipe pale yellow. smell almost absent or weakly farinaceous; taste mild to slightly bitter. spore print pale cream-coloured. spores (2.5-)2.8-4.5 x 2.0-3.5 μm, av. 3.0-4.2 x 2.3-3.4 μm, q 1.1-1.5(-1.8), qav. 1.2-1.4, in majority broadly ellipsoid or ellipsoid, some subglobose or oblong, smooth, not amyloid, in water colourless or with yellow, refractive body, in part also with brown granules (‘necropigment’), staining reddish in ammonia and koh, dark red to violet-brown in congored and acetocarmine. basidia 12-25 x 3.5-6.0 μm, clavate, 4-spored, in water with numerous brown granules and larger clots, staining like spores. lamella edge fertile. hymenophoral trama subregular, made up of rather narrow hyphae with cylindrical or slightly inflated elements, 13-90 x 3.0-14 μm, with numerous brown granules and clots. pileipellis mainly a poorly differentiated cutis, made up of repent, interwoven hyphae, 2.0-5.5 μm broad, with yellow intracellu34 e. arnolds lar pigment, towards the centre often with trichodermial fascicles of ascending and erect hyphae, often with ellipsoid to subglobose terminal cells up to 8.0 μm wide, with brown granules and clots. stipitipellis a cutis, made up of repent hyphae, 2.0-4.0 μm wide. clamp-connections absent. chemical reactions: all parts of the fresh basidiocarp turning immediately dark red-brown to violet-brown with koh 5% and 10% ammonia (macroscopically); preparations of all tissues turning immediately violet in koh 5% and reddish in 10% ammonia, also staining surrounding liquid; granules and clots becoming vinaceous red or dark violet. granules staining dark purple in acetocarmine with iron (seemingly siderophilous). habitat and distribution: saprotrophic, solitary or in small groups, on strongly decayed stumps or dead trunks of coniferous trees, mainly recorded from pinus sylvestris, also on pinus pinea and picea abies, in coniferous and mixed stands, mainly on acidic, sandy and loamy soils. july-nov. apparently widespread but rare in centraleurope; recorded from the netherlands, southern and eastern germany (l u d w i g 2001; w i l h e l m 2003), switzerland (k a l a m e e s 2004, as c. cerina), austria (k a l a m e e s 2004, as c. cerina), france (w i l h e l m 2003; b o n 1995) and italy. collections examined. austria: steiermark, gleichenberg, trautmannsdorf, 21 aug. 2003, w. wofar & a. hausknecht (wu, as calocybe cerina). – france: elsass, hardt, 16 sept. 1998, m. wilhelm 596 (herb. wilhelm); sondersdorf, 31 july 2000, m. wilhelm 709 (herb. wilhelm); oltingue, 27 aug. 2002, m. wilhelm 793 (herb. wilhelm). – italy: tuscany, orbetello, bosco di patanella, 27 oct. 1987, th. w. kuyper 2804 (l, as calocybe cerina). –– the netherlands: dwingeloo, lheederzand, 4 oct. 1990, e. arnolds 6100 (l, as calocybe cerina); smilde, hardersbosch, 13 nov. 1991, e. arnolds 6232 (l, as calocybe cerina); diever, state forest smilde, 3 nov. 1986, b. de vries 5193 (l, as calocybe spec.); diever, state forest smilde, 31 aug. 1992, j. baar 92/11 (l, as calocybe chrysenteron); ede, estate roekel, 21 sept. 2002, p.j. keizer s.n. (l, as calocybe cerina); bergen-schoorl, state forest, a.g. becker, 6 oct. 1990 (l, as calocybe cerina). callistosporium pinicola does not belong to the genus calocybe in view of the presence of small granules and larger clots of brown necropigment in the basidia, spores trama and pileipellis, as well as by the complete absence of clamp-connections. however, the necropigment can only be recognized as such in preparations in water (also of exsiccata), because it is immediately turning red to violet in ammonia and koh. since exsiccata are usually revived in these solutions the brown pigment is easily overlooked in dried specimens. moreover the necropigment is strongly staining purple with acetocarmine, thus mimicking the siderophilous granules of calocybe species. therefore it is not surprising that the majority of herbarium collections, identified as calocybe cerina, appears to belong to callistosporium pinicola. in this context it is interesting to note that m a i r e (1937) described callistosporium luteoolivaceum under the name tricholoma chrysenteron ( calocybe chrysenteron) var. olivascens maire (r e d h e a d 1982). apparently confusion between the two genera has occurred before. the presence of necropigment, the red staining of tissues in koh and ammonia and the absence of clamp-connections are all characters of the genus callistosporium sing. the present species belongs to subgenus callistosporium in view of the crowded lamellae, small, ellipsoid spores, short basidia and the pigment not turning blue a confusing duo: calocybe cerina and callistosporium pinicola (agaricales) 35 in ammonia (bon, 1991). taxonomy and nomenclature of this group are still rather controversial. bon (1991) distinguished two species, c. xanthophyllum (malenc. & bert.) m. bon and c. elaeodes (romagn. ex) m. bon, but r e d h e a d (1982) and n o o r d e l o o s (1995) united these taxa in one species and synonymised it with the north-american c. luteoolivaceum (berk. & curtis) sing, which name has priority in that case. l u d w i g (2001) described c. foetens e. ludwig as additional new species in this subgenus, characterized by a strong unpleasant smell. callistosporium pinicola differs from all species mentioned above in the first place by the very small spores. ludwig (2001) described the spores of c. luteoolivaceum as 5-5.5(-6.5) x 3.5-4 μm and b o n (1991, as c. xanthophyllum) as (5.0-)5.5-6.5(-7) x (3-)4-4.5(-5) μm. spores in the type collection of c. elaeodes measured 6.5-7.5 x 3.5-4.4(-5.0) μm (b o n 1976). in addition it is striking that the basidiocarps of c. pinicola are generally lacking green colours, so characteristic for other species of callistosporium. only in one of the studied collections a weak olivaceous tone was observed in the centre of the pileus. the habitat on stumps and trunks of coniferous trees is also characteristic for c. pinicola. other species are usually growing on deciduous wood or litter. in view of these characteristics the identity of the collection, depicted by l u d w i g (2000) in pl. 6.2 b as c. luteoolivaceum var. minor, is in my opinion doubtful since the pileus is distinctly green and the spores are described as larger: 4-5 x 3-4 μm. in the field callistosporium pinicola may also be mistaken for a species of simocybe or gymnopilus. the taxonomic position of calocybe juncicola next to calocybe cerina sensu arnolds & becker, there is a second european species of calocybe in which red staining of tissues in alkaline solutions has been reported, viz. calocybe juncicola (r. heim) sing. (m o r e n o 1995). since this reaction is considered a characteristic feature of the genus callistosporium, i have examined two collections of that species in order to re-evaluate its taxonomic position. calocybe juncicola (r. heim) sing. in sydowia 15: 47. (‘1961’) 1962. – fig. 3. tricholoma chrysenteron var. juncicola r. heim in treb. mus. cienc. nat. barcelona 15: 101. 1934; calocybe chrysenteron var. juncicola (r. heim) g. moreno in cryptog. mycol. 15: 240. 1995; rugosomyces chrysenteron var. juncicolus (r. heim) m. bon in doc. mycol. 21 (82): 65. 1991. sel. icon. – cetto, funghi vero 6: pl. 2414. 1989; ludwig, pilzkompendium 1: pl. 19, fig. 7.9. 2000. sel. descr. & figs. – hauskn. & zuccherelli in boll. gruppo micol. g. bres. 1994: 68-69, fig. 1 a-c. 1994; ludwig, pilzkompendium 1: 48. 2000; g. moreno in cryptog. mycol. 15: 240-241, fig. 1. 1995. pileus 10-30 mm, truncate campanulate to hemispherical at first, then convex to flattened, with or without umbo, often irregular, not hygrophanous, not striate, bright orange to golden yellow or brownish orange (e.g. k. & w. 4a8, 6a6, 6a7, 6b8), uniformly coloured or with slightly paler margin, slightly aerenchymatic in places when young, glabrous and dry. lamellae, l 30-40, l 3-7, emarginate to adnexed, moderately crowded to crowded, segmentiform, up to 4 mm broad, thin, vividly yellow, butter-yellow or ochre-yellow (k. & w. 4a5/6, 4b6), with concolor36 e. arnolds ous, entire or eroded edge. stipe 30-60 x 2-10 mm, subcylindrical or tapering towards base, often compressed or twisted, often slightly broader towards apex or base, solid, paler than the pileus, pale yellow, butter-yellow to brownish yellow (e.g. k. & w. 4a4, 5, 6, 4b6), fibrillose striate lengthwise, appearing pruinose by aerenchymatic tissue. context in pileus up to 4 mm thick, pale yellow. smell and taste strongly farinaceous, taste slightly adstringent. spore print unknown. spores (3.5-)4.5-6.5 x (2.5-)3.0-3.7(-4.0) μm, av. 5.0-5.4 x 3.2-3.3 μm, q 1.3-1.9, qav. 1.5-1.65, ellipsoid or ellipsoid-oblong, smooth, not amyloid, in water and ammonia colourless. basidia 25-36 x 5.0-8.0 μm, clavate, 4-spored or a few 2-spored, in part with few to fairly numerous small granules. lamella edge fertile, but sometimes with scattered cylindrical basidiola, 18-23 x 4.0-5.0 μm. pleurocystidia absent. hymenophoral trama subregular, made up of rather narrow hyphae with cylindrical or slightly inflated elements, 2.0-14 μm broad. pileipellis a poorly differentiated cutis, made up of repent, interwoven hyphae, 1.5-5.0 μm broad, in places with ascending to erect hyphae, with pale yellow intracellular pigment. stipitipellis a cutis, made up of repent hyphae, 2.0-4.0 μm wide. clamp-connections abundant in hymenium and trama. chemical reactions: preparations of all tissues turning immediately reddish to violet in koh 5% and ammonia 10%; macroscopic reactions on fresh basidiocarps unknown. granules in basidia staining dark purple in acetocarmine with iron (siderophilous). habitat and distribution: saprotrophic, usually fasciculate, on roots and litter in dense tussocks of juncus spp. (e.g. juncus maritimus, juncus acutus) in open forests and dune slacks. nov. apparently very rare in southern europe, known from italy and spain. fig. 3. calocybe juncicola. a. basidiocarps x 1; b. spores x 2000; c. basidia x 1000 (all from noordeloos 20266). a confusing duo: calocybe cerina and callistosporium pinicola (agaricales) 37 collections examined. – italy: ravenna, pineta san vitale, bardello, 6 nov. 2000, a. hausknecht et al. (wu no. 20752); same dat. and loc., m.e. noordeloos 20266 (l). the reddish staining of tissues in alkaline solutions was only reported by m o r e n o et al. (1995), who mentioned a violet discoloration of the solution when making a preparation of the lamellae in koh. the reaction was not mentioned in other descriptions (h a u s k n e c h t , z u c c h e r e l l i 1994; c e t t o 1989; l u d w i g 2001) or in the descriptive notes accompanying the collections. the reddening in ammonia and koh is very similar to that of callistosporium pinicola, although slightly weaker. nevertheless, calocybe juncicola seems not to be related to the latter species in view of the absence of brown necropigment, the abundance of clamp-connections and the presence of truly siderophilous granules in the basidia, although these granules seem to be less numerous and striking than in other calocybe species. otherwise, c. juncicola differs from c. chrysenteron (incl. var. cerina) mainly in duller colours, striate stipe, larger and more elongate spores and specific habitat. a second european species of calocybe with violaceous red discoloration of tissues in koh is rugososmyces pudicus m. bon & contu, collected in moist forest of populus on sardegna (c o n t u , b o n 2000). it has small basidiocarps with a pinkish brown to yellow-brown pileus up to 20 mm, differing mainly from c juncicola in immediate reddening of the context on exposure to the air and broader spores (5.0-6.0 x 3.5-4.5 μm) that are slightly rugulose, as was also demonstrated with sem microscopy (c o n t u , o r t e g a 2001). calocybe or rugosomyces? the genus calocybe was erected (but invalidly published) by k ü h n e r (1938) in order to accommodate tricholomoid, white-spored agarics with basidia containing siderophilous granules in combination with vividly coloured basidiocarps. donk (1962) validated this name and selected calocybe georgii (l.) kühner ( c. gambosa (fr.: fr.) donk) as type species. several authors have discussed the possible heterogeneity of this genus since. k a l a m e e s (1992) has proposed the genus tricholomella for calocybe constricta (fr.: fr.) sing., deviating from other species in presence of a partial veil, verrucose spores, large basidia and ecological preference for substrates with high ammonia content. this distinction was later confirmed by phylogenetic studies using molecular characters (h o f f s t e t t e r et al. 2002; m o n c a l v o et al. 2002). the genus gerhardtia was created by b o n (1994) for calocybe borealis a. riva ( lyophyllum incarnatobrunneum ew. gerhardt), a species deviating in minutely verruculose spores and absence of clamp-connections. since verruculose spores have also been observed in rugosomyces (calocybe) pudicus (c o n t u and o r t e g a 2001), the status of this genus seems questionable. the phylogenetic relationships with the rest of calocybe (and lyophyllum) has not yet been studied. earlier r a i t h e l h u b e r (1979) erected the genus rugosomyces for the two species of calocybe with a cellular pileipellis (hymeniderm) instead of a pileipellis of slender hyphae (cutis): the type species rugosomyces onychinus (fr.) raithelh. and r. fallax (sacc.) m. bon. this concept was adopted by e.g. h o r a k (2005), but bo n (1991) extended rugosomyces to all small species with collybioid habit, a pileipellis 38 e. arnolds with more or less free ending hyphal tips, often with inflated cells, and mixed parietal and intracellular pigments. calocybe was restricted by b o n (l.c.) to a few large, tricholomoid species with intracellular pigments. this concept was accepted by e.g. k a l a m e e s (2004). however, molecular studies of the lyophylleae have demonstrated that calocybe (with the exception of c. constricta, possibly also c. borealis; see above) forms a single clade and is probably of monophyletic origin (h o f f s t e t t e r et al. 2002; m o n c a l v o et al. 2002). therefore there seems to be no good reason to recognise rugosomyces as a separate genus. in this connection i propose two new combinations: calocybe pudica (m. bon & contu) arnolds comb. nov. basionym: rugososmyces pudicus m. bon & contu in doc. mycol. 29 (116): 35. 2000. calocybe obscurata (p. karst.) arnolds comb. nov. basionym: tricholoma cerinum pers. ssp. obscuratus p. karst. in meddeland. soc. fauna fl. fenn. 5: 43. 1879. callistosporium and calocybe the genus callistosporium is assigned to the tricholomatoideae and not to the lyophylloideae in view of the absence of siderophilous granules in the basidia. also in phylogenetic analysis the genera seem not to be closely related to each other (m o n c a l v o et al. 2002). however, it might be interesting to study the relationships between these groups more closely in view of the remarkable convergence between some species. also the chemical composition of the necropigment in callistosporium deserves attention. acknowledgements. this paper is dedicated with great pleasure to professor alina skirgiełło, a living monument of vitality and a source of inspiration for all of us! thanks are due to anton hausknecht (maissau, austria), markus wilhelm (allschwil, switzerland) and the curators of the herbaria of the botanical museum in copenhagen (c) and the national herbarium in leiden (l) for sending me valuable collections on loan. i am also indebted to thomas kuyper (wagenin gen, the netherlands) for some critical remarks on the manuscript of this paper. references a r n o l d s e., b e c k e r a. 1993. over calocybe cerina en enkele verwanten. coolia 36: 69 78. b o n m. 1976. le genre callistosporium singer. doc. mycol. 6 (22/23): 279 286. b o n m. 1991. les genres echinoderma (locq. ex bon) st. nov. et rugososmyces raithelhuber ss. lato. doc. mycol. 21 (82): 61 66. b o n m. 1991. flore mycologique d’ europe 2. tricholomataceae 1. doc. mycol., mém. hors sér. 2: 1 149. b o n m. 1994. deux lyophylloideae interessantes et le genre gerhardtia st. et nom. nov. doc. mycol. 24(93): 65 68. b o n m. 1999. flore mycologique d’ europe 5. les collybio marasmïoïdes et ressemblants. doc. mycol., mém. hors sér. 5: 1 171. c e t t o b. 1989. funghi vero 6. trento. c o n t u m., b o n m. 2000. une nouvelle espèce de rugosomyces ‘rougissant’. doc. mycol. 29(116) : 35 36. c o n t u m., o r t e g a a. 2001. studi sulle lyophyllaceae della sardegna.v. morfologia sporale di ru gososmyces pudicus bon & contu ed implicazioni sulla sua posizione sistematica. bol. soc. micol. madrid 26 : 173 176. a confusing duo: calocybe cerina and callistosporium pinicola (agaricales) 39 d o n k m.a. 1962. the generic names proposed for agaricaceae. beih. nova hedwigia 5 : 1 320. f r i e s e.m. 1821. systema mycologicum 1. gryphiswaldiae. h a u s k n e c h t a., z u c c h e r e l l i a. 1994. ritrovamenti interessanti dal ravennate. 3e parte. boll. gruppo micol. g. bresadola 1994: 67 95. h o f s t e t t e r v., c l é m e n c o n h., v i l g a l y s r., m o n c a l v o j m. 2002. phylogenetic analyses of the lyophylleae (agaricales, basidiomycota) based on nuclear and mitochondrial rdna sequences. mycol. res. 106: 1043 1059. h o r a k e. 2005. röhrlinge und blätterpilze in europa. elsevier, münchen. k a l a m e e s k. 1992. tricholomella, a new genus, with the distribution data of tricholomella constrictum comb. nov. in east europe and asia. persoonia 14: 445 447. k a l a m e e s k. 1995. on rugosomyces fallax and allied species (tricholomatales). doc. mycol. 25 (98 100): 229 236. k a l a m e e s k. 2004. palearctic lyophyllaceae (tricholomatales) in northern and eastern europe and asia. scripta mycologica 18. tartu. k o n r a d p., m a u b l a n c a. 1927. icones selectae fungorum 3. paris. k ü h n e r r. 1938. utilisation du carmin acétique dans la classification des agarics leucosporés. bull. mens. soc. linn. lyon 7: 204 212. l a n g e j.e. 1935. flora agaricina danica 1. copenhagen. l u d w i g e. 2000. pilzkompendium 1, abbildungen. ihw verlag, eching. l u d w i g e. 2001. pilzkompendium 1, beschreibungen. ihw verlag, eching. m a i r e r. 1937. fungi catalaunici. contribution à l’ étude de la flore mycologique de la catalogne. publ. inst. bot. barcelona 3 (4): 1 128. m o n c a l v o j m, v i l g a l y s r. , r e d h e a d s . a . , j o h n s o n j . e . , j a m e s t. y . , a i m e m . c . , h o f s t e t t e r v. , ve r d u i n s . j . w. , l a r s s o n e . , b a r o n i t. j . , t h o r n r . g . , j a c o b s s o n s . , c l é m e n c o n h . , m i l l e r jr., o.k. 2002. one hundred and seventeen clades of euagar ics. mol. phylogenet. evol. 23: 357 400. m o r e n o g., a r e n a l f., g o n z á l e z v. 1995. algunos agaricales de espana. crypt. mycol. 15: 239 254. n o o r d e l o o s m.e. 1995. callistosporium. in: bas, c., kuyper, th.w., noordeloos, m.e., vellinga e.c. (eds). flora agaricina neerlandica 3: 104. a.a. balkema, rotterdam, brookfield. p e r s o o n c.h. 1801. synopsis methodica fungorum. gottingae. r a i t h e l h u b e r j. 1979. calocybe kühner ein sammelgattung? metrodiana 8: 9 10. r e d h e a d s.a. 1982. the systematics of callistosporium luteo olivaceum. sydowia 35: 223 235. w i l h e l m m. 2003. callistosporium luteoolivaceum var. minor m. bon, kleinsporiger scheinrübling. schweiz. z. pilzk. 2003: 62 66. podwójna niejasność: calocybe cerina i callistosporium pinicola (agaricales) s t r e s z c z e n i e nazwa calocybe cerina była ostatnio stosowana dla dwóch różnych gatunków grzybów aga rikoidalnych. na podstawie analizy materiałów z różnych krajów europejskich autor dochodzi do wniosku, że prawdziwie gatunek calocybe cerina jest bardzo bliski c. chrysenteron i propo nuje nową kombinację calocybe chrysenteron var. cerina podając opis nowego taksonu. więk szość badanych kolekcji określonych jako c. cerina, okazała się taksonem, który bon opisał prowizorycznie jako callistosporium luteooliraceum fo. minor. w nieniejszym artykule został on opisany jako nowy gatunek: callistosporium pinicola. opis pozwala wyraźnie odróżnić inny gatunek calocybe juncicola. równocześnie dyskutowany jest status rodzajów rugosomyces i calocybe. podane są też nowe kombinacje calocybe obscurata i calocybe pudica. 2014-01-01t11:43:19+0100 polish botanical society clitocybe splendoides h.e. bigelow (agaricales, tricholomataceae), a new species for poland halina komorowska department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków k o m o r o w s k a h. clitocybe splendoides h.e. bigelow (agaricales, tricholomataceae), a new species for poland. acta mycol. 42 �2��1��-1�1, 2���.acta mycol. 42 �2��1��-1�1, 2���. records of clitocybe splendoides h. e. bigelow, a rare fungus belonging to the species group of the c. gibba type, associated with deciduous forests, especially beech forests, collected at three localities in poland are described. key words� agaricales, tricholomataceae, clitocybe splendens, c. splendoides, poland introduction clitocybe splendoides h.e. bigelow, lloydia 31� 4�. 1�6�. – c. splendens �fr.� gillet ss. bresadola, icon. mycol. iv� 16�. 1�2�. – c. infundibuliformis �fr.� ex schaef. var. splendens �bres. nec al.� kühner & romagnesi flore analytique� 13�. 1�53. systematic arrangement� b i g e l o w �1�6��decided to propose the epithet splendoides as nomen novum for c. splendens �fr.� gillet ss. bresadola, icon. mycol. 4� pl. 16�. 1�2� because “in the sense of most modern investigators, c. splendens is a synonym of c. gilva, c. inversa, or c. flaccida. while there are some disagreements in the literature on the limits of these three species, at least all have ornamented spores” �b i g e l o w 1�6�� 4��. these three species are now placed in the genus lepista, and l. splendens �pers.� gillet is either distinguished as a separate species �b o n 1���� or considered as a synonym of l. gilva or l. flaccida �k u y p e r 1��5; h o r a k 2��5�. c. splendens sensu bresadola has smooth, eliptical, obovate or pyriform spores in face view, and occurs on leaves under beech, trees; its nomen novum = c. splendoides h.e. bigelow. although bigelow initially placed it in the section infundibuliformes because of the shape of the pileus and the resemblance of the pileal colour to some colours occasionally found in c. gibba �fr.� p. kumm. �b i g e l o w 1�6�� 43�, he later recognised this disposition as incorrect. species close to c. gilva have encrusted pigments on the surface hyphae, and since such pigments are absent in c. splendoides, he placed it in the section candicantes, noticing that c. splendoides h. e. bigelow and c. catina �fr.� quél. may represent a possible link of the candicantes to acta mycologica vol. 42 �2�� 1��-1�1 2��� dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 1�� h. komorowska the infundibuliformes �b i g e l o w 1��2� 11��. other mycologists assign it to the new section gilvoideae harmaja �s i n g e r 1���� 2��, 21�; c l é m e n ç o n 1��4� 3��. b o n �1���� 3�� and h o r a k �2��5� 1�6, 1���, for instance, place it in the section clitocybe �= infundibuliformes fr.� with other species of the group c. gibba. selected descriptions� bigelow, beihefte nova hedw. �2� 11�. 1��2; bon, doc. mycol. 13�51�� 14. 1��3; clémençon, beihefte z. mycol. 5� 3�. 1��4; schwöbel, beiträge kenn. pilze mitteleuropas 1� �. 1��4; bon, doc. mycol. mémoire 4� 3�. 1���; urbonas, lietuvos grybai �/2� 33. 1���. iconography: bresadola 1�2�� 16�; bon 1���� pl. 1a; schwöbel 1��4� �, abb. 1 �as c. splendens ss. bresadola�; urbonas 1���� vi. 4e-h. many new rare and interesting species, including clitocybe splendoides, were found during investigations on the genus clitocybe. morphological characteristics pileus 3-� cm, plane, soon depressed, at last infundibuliform, margin straight or narrowly decurved, not striate, yellowish, oldest specimens beige-white, surface moist, context thin, whitish, odour present but undefined. lamellae forked, intervenose, thin, narrow, up to 5 mm, crowded, white edges even and straight, somewhat undulate in age. stipe 3-6 cm long, apex �.5-1.� cm broad, straight, cylindrical, whitish, surface glabrous, base with whitish tomentum. dry specimens with sunset pileus, 1�d4 to 1�d6, 1�e6, and whitish stipe, 1�b1, 1�b2 �m a e r z , p a u l 1�5��, lamellae seem slightly duller than stipe because of water-brown edges. in exsiccates pileus not staining deep chestnut brown or dark chocolate brown where treated with 5% koh (see first specimens in colour illustration, fig. 1�. hyphae of pileus cutis cylindric, 4.6-6.1 μm, of context 7.7-13.8 μm diam, hyaline in koh, membranal and intracelular pigments present. cystidia absent. basidia 20.4-28.1 × 4.1-6.1 μm, 4spored rarely 1 or 2-spored. spores 6.1-7.7 (8.2) × (2.8-) 3.1-4.3 μm, elipsoid to obovoid or pyriform, not amyloid (fig. 2). rare in deciduous or mixed forest, on beech leaves. august to october. examined specimens� 343. lvov upland, 343.2. the roztocze, „debry” reserve, near krasnobród, 5.��.2��4, leg. h. komorowska, kram-f 555��; 512. northern subcarpathian region, 512.3. brama krakowska gate, kraków, las wolski forest, 2�.1�.2��4, leg. j. lichoń, kram-f 55580; 522. outer eastern carpathians, 522.1. beskidy wschodnie mts., 522.12. bieszczady zachodnie mts., s. myczkowski, „hulskie” reserve, tilio-carpinetum, 2�.��.1��6, leg. h. komorowska, kram-f 555��. numbers and names of geographical regions follow k o n d r a c k i �2��1�. notes c. splendoides is reported in the literature only by specialists working on the genus clitocybe and is usually not distinguished from the very common c. gibba by other mycologists. only a very detailed examination of specimens under a microscope reveals that there exists a group of similar species �c. splendoides h.e. bigelow, c. glarosa reling & monthoux, c. costata küner & romagn., c. bresadoliana singer and c. gracilis �h.e. bigelow & a.h. sm.� harmaja or c. alnicola h.e. bigelow� 1�� h. komorowska it was very difficult to find any information on the distribution of c. splendoides in europe in the available literature. some data on the occurrence of this species were obtained in luxemburg �a n o n y m o u s 1���� ��1�, lithuania �u r b o n a s et al. 1��6� 1�; u r b o n a s 1���� 33� and germany �s c h w ö b e l 1��4� ��. there are also reports on collections from europe �without details, however� and the usa �b i g e l o w 1�6�, 1��2; b o n 1���; h o r a k 2��5�. b r e s a d o l a �1�2�� also reports this species from asia �siberia� and south africa. the anatomy of the pileus is very uniform in the genus clitocybe, therefore, as suggested by h a r m a j a �1�6�� 25�, only a detailed examination of the epidermis and subepidermis can help identify the species. the most reliable features are the shape and size of spores, the two being the most constant characters. bigelow emphasized the lack of incrustation of pileipellis hyphae in c. spendoides, a feature that is present in c. gibba and other species of its complex. in my opinion this feature is of lesser importance and the observations of the polish specimens lead me to believe that the shape and size of spores is the most significant character that distinguishes c. splendoides from closely related species. spores are mostly elongated, narrowly elliptical or elliptical, and very few are lacrimoid, similar to those of c.gibba but smaller. species belonging to the c. gibba complex will be described in a separate publication where the differences between them will be discussed in detail. acknowledgements. the author wishes to thank dr. anna ronikier for the english translation of a part of this paper and mr jacek lichoń for donating the specimen from kraków. the studies, supported by the state committee for scientific research (kbn grant po4g 024 22), were carried out in reserves and national parks in poland in the period between 2��2 and 2��4. references a n o n y m o u s 1���. rapport sur la session de la smf tenue à echteenach �grand-duché de luxembourg� du 2� september au 4 octobre 1���. bull. soc. mycol. fr. 1�6� 6�–��. b r e s a d o l a j . 1�2�. iconographia mycologica, vol. 4, societá botanica italiana, mediolani. b i g e l o w h . e . 1�6�. the genus clitocybe in north america. ii. section infundibuliformes. lloydia 31�1�� 43–62. b o n m . 1���. les clitocybes, omphales et ressemblants. tricholomataceae �2�, clitocyboideae. doc. mycol. mémoire hors 4� 1–1�1. c l é m e n ç o n h . 1��4. kompendium der blätterpilze� clitocybe. beih. z. mykol. 5� 1–6�. h o r a k e . 2005. röhrlinge und blätterpilze in europa. kleine kryptogamenflora iib/2. spectrum academischer verlag. k o n d r a c k i j . 2001. geografia regionalna polski (regional geography of poland). ed. 2. wydawnictwo naukowe pwn, warszawa. 441 pp. k u y p e r t. w. 1��5. clitocybe �fr.� staude, schwämme mitteldeutschl.� xxviii, 122. 1�5� �in�� c . b a s , t h . w. k u y p e r , m . e . n o o r d e l o o s , e . c . ve l l i n g a �eds�. flora aga-e l o o s , e . c . ve l l i n g a �eds�. flora agaricina neerlandica. 3. a. a. balkema/rotterdam/ brookfield: 42–62. m a e r z a , p a u l m . r e a 1�5�. a dictionary of color. ed. 2. mcgraw-hill book company, inc. new york, toronto, london. s c h w ö b e l h . 1��4. trichterlinge aus dem c. gibba – formenkreis �zur abgrenzung von c. costata, c. catinus und c. splendens ss. bres.�. beiträge zur kenntnis der pilze mitteleuropas i� 5–1�. s i n g e r r . 1978. keys for the identification of the species of agaricales ii. sydowia 31: 193–237. u r b o n a s v. 1���. tricholomatales 2. �in�� v. u r b o n a s �ed.�. mycota lithuaniae �/2, uab “valstiečiu laikraštis”, vilnius. 216 pp. u r b o n a s v. , k a l a m e e s k . , l u k i n v. 1986. conspectus florum agaricalium fungorum (agaricales s. l.� lithuaniae, latviae et estoniae. vilnius. 13� pp. clitocybe splendoides 1�1 clitocybe splendoides h.e. bigelow �agaricales, tricholomataceae�, nowy gatunek dla polski s t r e s z c z e n i e w czasie badań nad rodzajem clitocybe, prowadzonych w rezerwatach i parkach narodowych w latach 2��2-2��4 w ramach projektu badawczego kbn �grant po4g �24�, zebrano wiele nowych, rzadkich i interesujących gatunków. jednym z nich jest przedstawiony wyżej clitocybe splendoides. bigelow nadał tę nazwę znanemu z europy i stwierdzonemu w usa c. splendens sensu bresadola. uzasadniał to dużym zamieszaniem w literaturze europejskiej i traktowaniem tego taksonu przez różnych mikologów jako synonimu c. gilva, c. inversa lub c. flaccida gatunków z szorstkimi zarodnikami. wszystkie te trzy gatunki obecnie większość mikologów umieszcza w rodzaju lepista. ich ranga taksonomiczna też jest różna – są dobrymi gatunkami lub tylko synonimami. c. splendens sensu bresadola ma gładkie eliptyczne zarodniki, a jego nową nazwą jest od 1968 roku c. splendoides. jego pozycja w rodzaju clitocybe zmieniała się, był umieszczany w różnych sekcjach. jest gatunkiem rzadkim i odnotowywanym głównie przez specjalistów, bywa nie odróżniany od c. gibba, który jako dość pospolity nie jest szczegółowo badany. uchodzi uwadze zróżnicowanie zarodników (ich wielkość i kształt), a to one decydują o przynależności do danego taksonu, wraz z innymi cechami makroskopowymi czy choćby reakcją makrochemiczną skórki kapelusza traktowanej 5% koh. inne gatunki z kompleksu c. gibba zostaną przedstawione w odrębnej publikacji. fig. 1. dry specimens of c. splendoides (kram f 55578). scale bar = 2 cm. no reaction of the pileipellis with 5% koh (see top photograph, left specimen). 2014-01-01t11:46:07+0100 polish botanical society the occurrence of entomopathogenic fungi in soils from mid-field woodlots and adjacent small-scale arable fields cezary tkaczuk1, tomasz krzyczkowski1 and rudolf wegensteiner2 1 siedlce university of natural sciences and humanities department of plant protection, prusa 14, pl-08-110 siedlce; tkaczuk@uph.edu.pl 2university of natural resources and life sciences, department of forest and soil sciences georg mendel str. 33, a-1180 vienna, rudolf.wegensteiner@boku.ac.at tkaczuk c., krzyczkowski t., wegensteiner r.: the occurrence of entomopathogenic fungi in soils from mid-field woodlots and adjacent small-scale arable fields. acta mycol. 47 (2): 191–202, 2012. the aim of this study was to compare the species composition and the intensity of entomopathogenic fungi occurrence in the soil from mid-field woodlots and adjacent small farmlands. the study material consisted of soil samples taken from a mid-field woodlot and an adjacent small-scale arable field in three different localities in the vicinity of siedlce. entomopathogenic fungi were isolated from soil using two methods: the insect bait method and the selective medium. the comparative study showed that the soil from mid-field woodlots was characterized by a richer species composition of entomopathogenic fungi than of adjacent arable fields. a total of six fungal species representing the anamorphs of hypocreales (ascomycota) were isolated from the soil collected from mid-field woodlots: b. bassiana, b. brongniartii, m. anisopliae, m. flavoviride, i. farinosa and i. fumosorosea. the presence of only three species was reported in the farmland soil: b. bassiana, m. anisopliae and i. fumosorosea. this fact confirms the important role of semi-natural habitats as a source of biodiversity of entomopathogenic fungi in agricultural landscape. it was found that entomopathogenic fungi together formed more colony-forming units in the soil from arable fields than that of neighbouring mid-field woodlots. b. bassiana was the species of fungus which infected more bait insect larvae and formed significantly more colony-forming units (cfu) in the soil from mid-field woodlots than that of farmland in the localities studied, whereas the trend was the opposite in the case of i. fumosorosea and m. anisopliae. given the presence of entomopathogenic fungi in the farmland soil in the three test places together, it was found that i. fumosorosea was dominant in the soil from the two arable fields, where this fungus infected more g. mellonella larvae and formed significantly more cfus than the other species of fungi. m. anisopliae was the second most frequently isolated farmland species. key words: insect-pathogenic fungi, hypocreales, (ascomycota) anamorphs, soil, agricultural landscape acta mycologica vol. 47 (2): 191–202 2012 192 c. tkaczuk et al. introduction fungi are the largest group of microorganisms colonizing the soil environment after bacteria. among them are entomopathogenic species which play an important role as one of the natural factors limiting populations of typical soil insect pests that overwinter or pupate in the soil environment (ignoffo et al. 1978; ferron 1981; miętkiewski et al. 1994; bajan et al. 1995). it is estimated that about 90% of arthropods harmful to plants spend at least part of their life cycle in soil (gaugler 1988). specimens which have died in soil as a result of mycoses spread diseases in the soil environment. the occurrence, development and pathogenicity of entomopathogenic fungi in soil are conditioned by a number of biotic and abiotic factors in the environment, as well as the agricultural and non-agricultural human activity. arthropods play a very important role among the biotic factors, as being potential hosts for these fungi, they determine their occurrence and survival in soil. the commonly accepted belief is that farmers affecting the physical and chemical properties of the environment, transforming the structure of vegetation both in fields and landscapes, contribute to the depletion of biological resources (ryszkowski 1985; karg 1989; ryszkowski, karg 1991). issues related to the protection of biodiversity in rural areas are among the most important priorities set by the european union in matters relating to environmental protection. agri-environment schemes are an essential tool for the protection of valuable natural habitats associated with agricultural landscape. in this landscape, farmland are surrounded by semi-natural habitats such as mid-field woodlots and field margins which are specific refugia for flora and fauna not present in arable fields (marshall, moonen 2002; meyling 2005). these habitats are the ideal place for many wintering arthropod species whose rich communities in turn determine a large variety and ease of propagation of pathogens, including entomopathogenic fungi. therefore, unfarmed parts of natural vegetation clusters marginal among arable fields, water tanks and trees introduced in strips or on small areas of land become very important for the preservation of the biotic assemblages. while the botanical, avifaunistic and entomological characteristics of such structural elements of agricultural landscapes are relatively well developed, data relating to entomopathogens that are contained therein are quite sporadic (bałazy, cysewski 2003; bałazy 2007). it is known, however, that the species composition of entomopathogenic fungi in semi-natural habitat soils, i.e. excluded from agrotechnical pressure, is different from that observed in soils intensively used for agriculture (steenberg 1995; bidochka et al. 1998; klingen et al. 2002; meiling, eilenberg 2006; tkaczuk 2008). the aim of this study was to compare the species composition and the intensity of entomopathogenic fungi occurrence in the soil from mid-field woodlots and adjacent small farmlands. the occurrence of entomopathogenic fungi 193 material and methods sampling and characterization of soil samples.the study material consisted of soil samples taken at the beginning of december 2006 in three localities in the vicinity of siedlce: purzec, borki siedleckie and pruszyn. in each place soil samples were collected from two environments: a mid-field woodlot and an adjacent arable field. winter crops were grown in the fields, and their areas in different places were respectively 1.0, 1.2 and 2.0 ha. mid-field woodlots were characterized by the following species composition of trees and shrubs: 1 purzec – trees on the area of about 500 m² with birch and pine dominance 2 borki siedleckie – trees on the area of about 800 m² with birch and pine dominance 3 pruszyn – trees on the area of about 1200 m² with the dominance of poplar and oak. samples were taken at random, separately for each environment from 10-15 points on the test area. samples from arable fields were taken at a distance of not less than 50 meters from the edge of trees. the material was collected using egner’s stick to a depth of 15 cm. a mixed sample was prepared from the collected material and stored in plastic bags at 0-4°c. immediately before starting the experiment in the laboratory, the soil was sieved and dried up to a suitable moisture content, optimal for fungal growth and limiting the development of nematodes. soil samples from all points were analysed to assess their granulometric composition (soil type), ph and organic carbon content. isolation of fungi. entomopathogenic fungi were isolated from soil using two methods: the insect bait method proposed by zimmermann (1986) and the selective medium method developed by strasser et al. (1996) and commonly used for the isolation of entomopathogenic fungi from soil (keller et al. 2003). the greater wax moth (galleria mellonella l.) was used as a bait insect. ten plastic boxes with a capacity of about 200 ml were filled with soil from each environment. ten galleria mellonella larvae were put in each box, a total of 100 larvae in the soil from each environment. the boxes of soil were placed in an incubator at 20-22°c. the first mortality control was conducted 7 days after the start of the experiment and then at 3-day intervals until the death of all larvae. dead larvae were washed in distilled water and then surface-sterilized for 30 seconds in 1% sodium hypochlorite solution. after rinsing twice in distilled water, the larvae were put on petri dishes with moistened filter paper. the dishes with the larvae were kept at 20-22°c in the dark. fungi growing on insects were transferred on standard media and then examined macroscopically. identification of isolated fungal species was done by using standard methods described by goettel and inglis (1997). fungal nomenclature follows sung et al. (2007) and index fungorum (www.indexfungorum.org). additionally, the concentration of colony-forming units (cfu) of entomopathogenic fungi in soil environments tested was determined applying a selective medium developed by strasser et al. (1996). for this purpose, 2 g of soil were weighed out from each mixed sample originating from a given point, and then 18 ml of distilled water were added with the addition of 0.05 triton x-100, and the components were vigorously shaken for about 35 194 c. tkaczuk et al. seconds. then 0.1 ml of the soil solution was poured out and spread using a glass spatula on a selective medium in four petri dishes which were the replicates. the dishes were placed in incubators at 22° c and after 8-10 days colonies of individual fungal species were counted. the results were expressed as the number of colonyforming units (cfu) of entomopathogenic fungi in 1 g of the soil. the selective medium used had the following composition: 10 g of peptone, 20 g of glucose and 18 g of agar were added to 1 litre of water. after sterilization and cooling, the following components were added to the medium: 0.6 g of streptomycin sulfate, 0.05 g of chlortetracycline, 0.05 g of cycloheximide and 0.1 g of dodine. the results were analysed statistically by performing a 2-factorial analysis of variance. a detailed comparison of the environments was made using the tukey’s test at significance level α = 0.05. results the comparative study of the composition and intensity of entomopathogenic fungi in the soil from mid-field woodlots and adjacent small-scale farmlands, conducted in 2006 in three localities in the vicinity of siedlce showed that the soil from mid-field woodlots was characterized by a richer species composition of these fungi. a total of six fungal species representing the anamorphs of hypocreales (ascomycota) were isolated from the soil collected from mid-field woodlots using g. mellonella larvae: beauveria bassiana (bals.-criv.) vuill , b. brongniartii (sacc.) petch, metarhizium anisopliae (metschn.) sorokin, m. flavoviride w. gams & rozsypal , isaria farinosa (holmsk,) fr. and i. fumosorosea wize. the presence of only three species was recorded in the farmland soil: b. bassiana, m. anisopliae and i. fumosorosea (tabs 1 and 2). in addition, apart from typical insect-pathogenic fungi, some accompanying fungal species with unproved entomopathogenic abilities were isolated from investigated soils by means of galleria bait method. the occurrence of fungi from the following genera was observed within this group: aspergillus p. micheli, fusarium link, gliocladium corda and mucor p. micheli ex l. the use of selective media allowed the isolation of only three species of entomopathogenic fungi from soils of both investigated habitats, such as b. bassiana, i. fumosorosea and m. anisopliae (tab. 3). four species of entomopathogenic fungi were isolated by means of bait insects from the soil samples taken from a mid-field woodlot in purzec: b. bassiana, m. anisopliae, m. flavoviride and i. fumosorosea. the fungus b. bassiana strongly dominated in this soil, and it infected 52.7% of g. mellonella larvae and formed 4.5 x 10³ colony-forming units (cfu) in 1 gram of soil. m. anisopliae and i. fumosorosea were of a mild intensity, infecting 6.3% and 1.1% of larvae and forming 1.7 and 1.5 x 10³ cfu gˉ¹. using the insect bait method only two species of entomopathogenic fungi were isolated from the adjacent farmland soil: m. anisopliae and i. fumosorosea that infected 96% and 1.0% of larvae (tab. 1). the fungus m. anispoliae also developed the most numerous colony-forming units in the farmland soil in purzec (15.0 x 10³ cfu gˉ¹), and b. bassiana 1.2 x 10³ cfu gˉ¹ (tab. 3). the occurrence of entomopathogenic fungi 195 it was found that entomopathogenic fungi invaded significantly more g. mellonella larvae and also formed more colony-forming units in the soil from the farmland than in the soil from the adjacent woodlot in this village – respectively 97.0 and 61.2%, and 16.2 x 10³ gˉ¹ and 7.7 x 10³ cfu gˉ¹. the soil sampled from mid-field woodlots in borki siedleckie was dominated by m. anisopliae which formed 11.8 x 10³ cfu in 1 gram of soil and infected 71% of larvae inserted into the soil. the fungus b. bassiana was also present on 23% of larvae (1.8 x 10³ cfu gˉ¹), while i. fumosorosea infected only 1% of bait insects and formed 1.2 x 10³ cfu gˉ¹. the same fungal species were isolated from the adjacent farmland soil but their dominance was different. the highest mortality of g. mellonella larvae was caused by the fungus i. fumosorosea (79.8%), and other species such as b. bassiana and m. anisopliae invaded a small number of larvae, respectively, 8.1 and 1.0%. the fungus i. fumosorosea also formed the most numerous colony-forming units in the farmland soil in borki siedleckie (9.7 x 10³ gˉ¹). it was found that entomopathogenic fungi infected more bait insect larvae and formed more colony-forming units in the woodlot soil than in the farmland soil in the village. the soil collected from the mid-field woodlot in pruszyn was found to have the richest entomopathogenic fungal species composition among all habitats studied. up to five fungal species occurred in that soil: b. bassiana, b. brongniartii, m. anisopliae, i. farinosa and i. fumosorosea. the fungus b. bassiana was the most abundant among them and it infected 65.0% of bait insect larvae and formed 9.5 x 10³ cfu in 1 gram of soil. other species caused the mortality of individual larvae from 1.0 to 3.0%, and formed a small quantity of colony-forming units from 0.3 to 2.0 x 10³ cfu in 1 gram of soil. only two fungal species were isolated using bait insects in the adjacent farmland soil. the fungus i. fumosorosea definitely dominated there and it table 1 mortality of galleria mellonella larvae (%) caused by entomopathogenic fungi in soils from mid-field woodlots and adjacent small-scale arable fields factor of mortality locality purzec borki siedleckie pruszyn a* b a b a b entomopathogenic fungi b. bassiana 52.7 0 23.0 8.1 65.0 0 b. brongniartii 0 0 0b 0 3.0 0 m. anisopliae 6.3 96.0 71.0 1.0 1.0 3.0 m. flavoviride 1.1 0 0 0 0 0 i. farinosa 0 b 0 0 0 1.0 0 i. fumosorosea 1.1 1.0 1.0 79.8 2.0 86.0 total 61.2 97.0 95.0 88.9 72.0 89.0 fungi of unproved entomopathogenic abilities mucor sp. 5.2 1.0 0 5.0 5.0 0 gliocladium sp. 3.1 0 0 2.0 1.0 2.0 fusarium sp. 2.1 1.0 1.0 0 2.0 0 aspergillus sp. 1.1 0 2.0 0 4.0 0 unsporulated mycelium 7.4 1.0 1.0 1.0 4.0 4.0 total 18.9 3.0 4.0 8.0 16.0 6.0 other causes nematodes 7.4 0 0 0 2.0 3.0 unidentified causes 9.5 0 1.0 3.0 10.0 2.0 alive specimens 3.1 0 0 0 0 0 abbreviations: * a – mid-field woodlot, b – arable field 196 c. tkaczuk et al. caused the death of 86.0% of larvae. moreover, the presence of m. anisopliae was observed on 3.0% of g. mellonella larvae (tab. 1). the fungus i. fumosorosea also formed by far the most numerous colony-forming units in the soil (13.2 x 10³ gˉ¹). m. anisopliae was also in a relatively high density (8.0 x 10³ gˉ¹), while b. bassiana formed the fewest colony-forming units (tab.3). comparing the occurrence of entomopathogenic fungi in the soil from woodlots to their neighbouring farmlands in the three villages studied, we can conclude that b. bassiana infected most bait insect larvae in woodlot soil (a total of 47.1% of larvae) (tab. 2). this fungus also formed the most numerous colony-forming units in two of the three mid-field woodlots studied. m. anisopliae was another species which infected most g. mellonella larvae in the soil under trees (26.1%). the fungus formed by far the most numerous colony-forming units in the woodlot soil in borki siedleckie. table 2 mortality of galleria mellonella larvae (%) caused by entomopathogenic fungi in soils from mid-field woodlots and adjacent small-scale arable fields (average from three localities) fungal species habitat arable field mid-field woodlot b. bassiana 2.7 47.1 b. brongniartii 0 1.0 m. anisopliae 33.3 26.1 m. flavoviride 0 0.4 i. farinosa 0 0.3 i. fumosorosea 55.5 1.4 total 91.5 76.3 table 3 number of colony forming-units of entomopathogenic fungi in the soil (cfu x 103 g-1) from mid-field woodlots and adjacent small-scale arable fields fungal species locality purzec borki siedleckie pruszyn a* b a b a b b. bassiana 4.5a** 1.2b 1.8b 0.5c 9.5a 1.3c m. anisopliae 1.7c 15.0a 11.8a 1.7c 0.3c 8.0a i. fumosorosea 1.5c 1.2c 9.7a 2.0c 13.2a total 7.7b 16.2a 14.8a 11.9ab 11.8b 22.5a abbreviations: * a – mid-field woodlot, b – arable field; ** values in lines followed by the same letters are not significantly different table 4 characteristics of soil samples locality habitat soil ph (ph w mol × dm³ kcl) soil type organic carbon content (%) borki siedleckie arable field 5.43 loamy sand 0.851 mid-field woodlots 4.78 sandy loam 2.477 purzec arable field 4.72 weakly loamy sand 0.686 mid-field woodlots 4.60 weakly loamy sand 1.114 pruszyn arable field 4.49 weakly loamy sand 0.538 mid-field woodlots 4.64 loamy sand 3.335 the occurrence of entomopathogenic fungi 197 given the presence of entomopathogenic fungi in the farmland soil in the three test places together, it was found that i. fumosorosea was dominant in the soil from the two arable fields. in the farmland soil from borki siedleckie and pruszyn, the fungus infected more g. mellonella larvae and formed significantly more cfus than the other species of fungi. m. anisopliae was the second most frequently isolated farmland species. this fungus definitely dominated bait insect larvae in the farmland soil in purzec, and it formed the most numerous colony-forming units in 1 gram of soil (tabs 1 and 3). figure 1 illustrates the difference in density of colony-forming units of entomopathogenic fungi in soils from arable fields and adjacent woodlots. it was found that entomopathogenic fungi together formed more colony-forming units in the soil from arable fields than that of neighbouring mid-field woodlots. the fungus b. bassiana formed significantly more colony-forming units (cfu) in the soil from mid-field woodlots than that of farmland in the localities studied, whereas the trend was the opposite in the case of i. fumosorosea and m. anisopliae (fig. 1). discussion transformation of landscape relations, especially deforestation for agricultural production and the simplification of species composition and the age structure of forest stands lasting for at least two centuries are not conducive to the maintenance of species-rich groups of insects and their pathogens in anthropogenically transformed environments (altieri 1999). for these reasons, unfarmed parts of natural vegetation clusters, marginal among arable fields, (field margins, roadsides, swamps), water tanks, as well as trees and shrubs introduced in strips on small areas of land become fig. 1. number of colony forming-units of entomopathogenic fungi in the soil (cfu x 103 g-1) from mid-field woodlots and adjacent small-scale arable fields (average from three localities) * values followed by the same letters are not significantly different 198 c. tkaczuk et al. very important for the preservation of the biotic assemblages (bałazy, cysewski 2003). these habitats are specific refugia for flora and fauna which are not present in farmland (marshall, monen 2002), and are the ideal place for many wintering species of arthropods. it was found that in the area of semi-natural strips with a width of 3 to 6 m on the edges of fields, the number of living species of arthropods is more than two times higher than in their neighbouring farmland (meek et al. 2002). according to karg et al. (2003), the biomass of dipteran larvae in the soil from mid-field woodlots in turew was 80-fold higher than in the soil from the adjacent field, and the total biomass of macrofauna was 90-fold higher. with regard to mesofauna, the biomass was 9-fold higher in the shelterbelt soil than in the soil from the adjacent field. the species richness of arthropods, especially insects, in the area of mid-field woodlots undoubtedly affects the species diversity of their natural enemies, including entomopathogenic fungi. this was confirmed in our research. the species composition of entomopathogenic fungi was far richer in the soil from mid-field woodlots than in the soil from neighbouring farmlands. in the soil collected from woodlots, a total of six fungal species were isolated using g. mellonella larvae: b. bassiana, b. brongniartii, m. anisopliae, m. flavoviride, i. farinosa and i. fumosorosea. the presence of only three species was reported in the soil from arable fields: b. bassiana, m. anisopliae and i. fumosorosea. chandler et al. (1997), meyling and eilenberg (2006) and tkaczuk (2008) also draw attention to the richer species composition of entomopathogenic fungi in soils from shelterbelts surrounding arable fields, and emphasize the important role of natural habitats as specific refugia for the survival of these fungi in agrocenoses. mayling (2005) showed, moreover, that shelterbelts around arable fields are also reservoirs for a variety of phylogenetic lines of the fungus b. bassiana. studying strains of this species, obtained from a field of about 28 ha and an adjacent belt of trees with a total area of only 0.44 ha, he recorded a significantly higher genetic diversity of fungal isolates from under the trees than from the field where there was, in principle, only one genetic group of b. bassiana. according to the above-mentioned author, this fact confirms the important role of semi-natural habitats as a source of biodiversity of entomopathogenic fungi in agricultural landscape. b. bassiana was the species of fungus which infected most bait insect larvae and formed the biggest number of cfus in the soil from mid-field woodlots studied. this fungus is cited as co-dominant with i. fumosorosea in soils of semi-natural habitats in poland (tkaczuk 2008). according to vannninen (1995) and steenberg (1995), b. bassiana is characteristic of natural habitats (mid-field woodlots, shelterbelts) and is less common in agricultural soils. one of the concepts explains the dominance of b. bassiana in soils from mid-field woodlots which, as revealed by the study (tab.4) are much richer in organic matter than farmland soils, is the ability of this fungus to grow in the saprophagous phase (müller-kögler, zimmermann 1986; storey et al. 1989). b. bassiana definitely dominates in soil and forest litter, i.e. the agricultural landscape environments that are probably the most similar to woodlots. it should also be noted that the survival of entomopathogenic fungi in soil largely depends on the stability of environmental conditions and the related continuous or at least frequent presence of potential insect hosts. in its the occurrence of entomopathogenic fungi 199 development cycle, the species b. bassiana seems to use a strategy that ewald (1983, 1995) has called “sit and wait”, which means that the population growth and survival of the fungus in soil depend mainly on periodical infections of hosts, and factors limiting the population of hosts seem to substantially affect the survival of b. bassiana in the soil environment. wojciechowska et al. (1977), daoust & pereira (1986) and steenberg (1995) showed that the constant presence of arthropods in soil as potential hosts for b. bassiana has a significant impact on the survival of the species. given the presence of entomopathogenic fungi in the farmland soil in the three villages studied, it was found that i. fumosorosea was dominant in the soil from two arable fields, and m. anisopliae in one of the farmlands. as demonstrated by the analyses, all soils from the three fields studied were sandy loam (tab.4). according to tkaczuk (2008) the presence of entomopathogenic fungi in farmland soils in poland was dependent on soil type. sandy soils were dominated by i. fumosorosea and m. anisopliae, while i. fumosorosea, m. anisopliae and b. bassiana were of similar intensity in clay soils, whereas b. bassiana and then m. anisopliae were by far the most common in organic soils. the dominance of i. fumosorosea and m. anisopliae in farmland sandy soils is confirmed by earlier studies conducted by tkaczuk & miętkiewski (1996) and kleespies et al. (1989). these authors also report a very low intensity of b. bassiana in these soils. examining the concentration of colony-forming units of entomopathogenic fungi in the soil from large-scale fields and neighbouring strips of trees in austria and poland, tkaczuk (2008) showed that these fungi formed more colony-forming units in the soil under shelterbelts. in our study, in the case of soil from two of the three pairs of test environments (field-woodlots), there was a greater concentration of cfus of entomopathogenic fungi in the farmland soil. the differences may be due to the fact that our study was conducted in small-scale fields (up to 2 ha) where farmers use minimal chemical protection, while research conducted by tkaczuk (2008) related to fields with an area of more than 5 ha where extensive chemical treatments were applied (this is particularly true of research conducted in austria), which could adversely affect the development of entomopathogenic fungi in the soil. according to karg and bałazy (2009), the simplification of the structure of crops, chemical control of pests and cyclically repeated tillage practices aimed at the intensification of crops directly or indirectly eliminate pathogens of arthropods, on a par with parasites and predators, as trophic groups with specific environmental and nutritional requirements. however, most of them are characterized by a high adaptability to adverse environmental conditions, and usually a high reproductive and migration potential, they can quickly colonize the environments from which they have been eliminated as a result of agricultural practices, as long as there are suitable refugia around the fields. among such refugia can be undoubtedly small mid-field woodlots which are characteristic of the agricultural landscape in the nizina południowopodlaska lowland where this study was conducted. according to marshall & monen (2000) and meek et al. (2002), especially the transition area (ecotone) between a cultivated field and a stable semi-natural environment is a habitat for species characteristic of both environments, and it makes it possible to enrich depleted entomofauna of farmland. 200 c. tkaczuk et al. conclusions the occurrence, development and pathogenicity of entomopathogenic fungi in soil are conditioned by a number of biotic and abiotic factors in the environment, as well as the agricultural and non-agricultural human activity. in agricultural landscape, arable fields are surrounded by semi-natural habitats such as mid-field woodlots which are specific refugia for flora and fauna not present in arable fields. our comparative study showed that the soil from mid-field woodlots was characterized by a richer species composition of entomopathogenic fungi than of adjacent arable fields. a total of six fungal species representing the anamorphs of hypocreales (ascomycota) were isolated from the soil collected from mid-field woodlots: b. bassiana, b. brongniartii, m. anisopliae, m. flavoviride, i. farinosa and i. fumosorosea. the presence of only three species was reported in the farmland soil: b. bassiana, m. anisopliae and i. fumosorosea. this fact confirms the important role of semi-natural habitats as a source of biodiversity of entomopathogenic fungi in agricultural landscape. aknowledgements. the study was supported by the scientific and technological cooperation project: austria-poland (pl 12/2006); university of natural resources and life sciences, vienna (austria) and siedlce university of natural sciences and humanities (poland). references altieri m.a. 1999. the ecological role of biodiversity in agroecosystems. agric. ecosyst. environ. 74: 19–31. bajan c., kmitowa k., mierzejewska e., popowska-nowak e., miętkiewski r., górski r., miętkiewska z., głowacka b. 1995. występowanie grzybów owadobójczych w ściółce i glebie borów sosnowych w gradiencie skażenia środowiska leśnego. 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(effect of landscape structure on the occurrence of agrophagous pests and their antagonists). prog. plant prot./post. ochr. roślin 49 (3): 115–134. karg j., kajak a., ryszkowski l. 2003. impact of young shelterbelts on organic matter content and development of microbial and fauna communities of adjacent fields. pol. j. ecol. 51 (3): 283–290. keller s., kessler p., schweizer c. 2003. distribution of insect pathogenic soil fungi in switzerland with special reference to beauveria brongniartii and metarhizium anisopliae. biocontrol 48: 307–319. kleespies r., bathon h., zimmermann g. 1989. untersuchungen zum natürlichen vorkommen von entomopathogenen pilzen und nematoden in verschiedenen böden in der umgebung von darmstadt. gesunde pflanzen 41: 350–355. klingen i., eilenberg j., meadow r. 2002. effects of farming system, field margins and bait insect on the occurrence of insect pathogenic fungi in soils. agric. ecosyst. environ. 91: 191–198. marshal e.j.p., moonen a.c. 2002. field margins in northern europe: their functions and interactions with agriculture. agric. ecosyst. environ. 89: 5–21. meek b., loxton d., sparks t., pywell r., picket h., nowakowski m. 2002. the effect of arable field margin composition on invertebrate biodiversity. biol. conser. 106: 259–271. meyling n.v. 2005. population ecology and genetic diversity of entomopathogenic fungi (ascomycota: hyphocreales) in agroecosystems and field margins. ph.d. thesis, the royal veterinary and agricultural university, copenhagen meyling n.v., eilenberg j. 2006. occurrence and distribution of soil borne entomopathogenic fungi within a single organic agroecosystem. agric. ecosyst. environ. 113: 336–341. miętkiewski r., tkaczuk c., żurek m., l.p.s. van der geest 1994. temperature requirements of four entomopathogenic fungi. acta mycol. 29 (1): 109–120. müller-kögler e., zimmermann g. 1986. zur lebensdauer von beauveria bassiana in kontaminiertem boden unter freilandund laboratoriumsbedungungen. entomophaga 31: 285–292. ryszkowski l. 1985. impoveriso agriculthment of soil fauna due the agriculture. 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(in:) t.a. jackson, t.r. glare (eds). microbial control of soil dwelling pests. agresearch, lincoln, new zeland: 125–130. sung g.h., hywel-jones n.l., sung j.m., luangsa-ard j.j., shrestha b., spatafora j.w. 2007. phylogenetic classification of cordyceps and the clavicipitaceous fungi. stud. mycol. 57: 5–59. tkaczuk c. 2008. występowanie i potencjał infekcyjny grzybów owadobójczych w glebach agrocenoz i środowisk seminaturalnych w krajobrazie rolniczym. (occurrence and infective potential of entomopathogenic fungi in the soils of agrocenoses and semi-natural habitats in agricultural landscape). rozprawa naukowa nr 94. wyd. akademii podlaskiej, siedlce, 160 pp. tkaczuk c., miętkiewski r. 1996. occurrence of entomopathogenic fungi in different kinds of soil. rocz. nauk rol., seria e 25(1/2): 44–48. vänninen i. 1995. distribution and occurrence of four entomopathogenic fungi in finland: effect of geographical location, habitat type and soil type. mycol. res. 100: 93–101. 202 c. tkaczuk et al. wojciechowska m., kmitowa k., bajan c. 1977. the effect of carbamidae herbicides linuron and monolinuron on the three of species of entomopathogenic fungi. ii pot experiments. pol. ecol. stud. 3 (2): 43–57. zimmermann g. 1986. galleria bait method for detection of entomopathogenic fungi in soil. j. app. entomol. 2: 213–215. występowanie grzybów entomopatogenicznych w glebach z zadrzewień śródpolnych i sąsiadujących z nimi małoobszarowych pól uprawnych streszczenie celem podjętych badań było porównanie składu gatunkowego i nasilenia występowania grzybów entomopatogeniczych w glebach z zadrzewień śródpolnych i sąsiadujących z nimi pól uprawnych. materiał do badań stanowiły próby gleby pobrane w trzech miejscowościach w okolicach siedlce z dwóch środowisk: zadrzewienia śródpolnego i sąsiadującego z nimi małoobszarowego pola uprawnego. grzyby owadobójcze izolowano z gleby stosując metodę owadów pułapkowych (z użyciem larw galleria mellonella) oraz podłoże selektywne. przeprowadzone badania porównawcze wykazały, że gleby spod zadrzewień śródpolnych charakteryzowały się bogatszym składem gatunkowym tych grzybów. z gleby pobranej spod zadrzewień śródpolnych na larwy g. mellonella wyizolowano w sumie sześć gatunków grzybów, były to: b. bassiana, b. brongniartii, m. anisopliae, m. flavoviride, i. farinosa i i. fumosorosea. w glebie z pól odnotowano obecność tylko trzech gatunków: b. bassiana, m. anisopliae i i. fumosorosea. fakt ten potwierdza istotną rolę środowisk seminaturalnych jako źródła bioróżnorodności grzybów owadobójczych w krajobrazie rolniczym. stwierdzono że, grzyby entomopatogeniczne łącznie tworzyły więcej jednostek infekcyjnych (cfu) w glebie z pól uprawnych niż sąsiadujących z nimi zadrzewień śródpolnych. stwierdzono, że grzyb b. bassiana zainfekował więcej larw owada pułapkowego i tworzył istotnie więcej jednostek infekcyjnych w glebach z zadrzewień śródpolnych niż pól w badanych miejscowościach, natomiast w przypadku i. fumosorosea i m. anisopliae odnotowano tendencję odwrotną. biorąc pod uwagę występowanie grzybów entomopatogennych w glebie z pól uprawnych w trzech badanych miejscowościach łącznie, stwierdzono, że w glebie z dwóch pól uprawnych dominował i. fumosorosea. w glebach tych zainfekował on więcej larw g. mellonella i tworzył istotnie więcej jednostek cfu niż pozostałe gatunki grzybów. drugim najczęściej izolowanym gatunkiem z pól uprawnych był m. anisopliae. 2014-01-02t12:15:25+0100 polish botanical society arcangeliella scissilis, a rare american semihypogeous fungus francisco d. calonge1 and milagro mata2 1real jardín botánico, c.s.i.c., plaza de murillo, 2, s-28014 madrid, calonge@rjb.csic.es 2instituto nacional de biodiversidad, a.p., cri-22-3100, santo domingo, heredia, mmata@inbio.ac.cr calonge f.d., mata m.: arcangeliella scissilis, a rare american semihypogeous fungus. acta mycol. 48 (1): 21–25, 2013. as a consequence of the finding of arcangeliella scissilis in costa rica, we thought to carry out a full description of this material, giving some aspects on its taxonomy, ecology and distribution. key words: arcangeliella, russulales, astrogastraceous, costa rica introduction the study of the hypogeous russulales has been a subject of great interest for the senior author in the last years (moreno-arroyo et al. 1998a, b; calonge, vidal 1999, 2001; moreno-arroyo et al. 1999; calonge, martín 2000; vidal et al. 2002). the presence of arcangeliella scissilis in costa rica has been reported a few years ago (calonge, mata 2005), and now we are trying to give some new information on this very rare species. material and methods central america. costa rica: guanacaste, bagaces, área de conservación arenal, sector finca rio naranjo, collection consisted in 18 basidiomes, which appeared closely aggregated being difficult to separete each other, due to be partially melted at the base (fig. 1). no specific smell or taste was appreciated in fresh. they were growing semihypogeous in riparian forest soil, under quercus sp., 24-v-1999, leg. isaac lópez. inb 1545920. after the field observation, the material was studied in the laboratory under the light microscope, using 3% koh as liquid medium and observed in a nikon acta mycologica vol. 48 (1): 21–25 2013 doi: 10.5589/am.2013.003 22 f. d. calonge and m. mata labophot. for sem a spore deposit was coated with gold and observed in a hitachi s-3000n, belonging to real jardín botánico of madrid. results arcangeliella scissilis zeller & c.w. dodge, ann. mo. bot. gard. 22: 369 (1935) ≡ martellia scissilis (zeller & c.w. dodge) singer & a.h. sm., mem. torrey bot. club 21: 45 (1960). ≡ zelleromyces scissilis ((zeller & c.w. dogge) trappe, t. lebel & castellano, mycotaxon 81: 205 (2002). = hydnangium gilkeyae zeller & c.w. dodge ann. mo. bot. gard. 22: 371 (1935). = zelleromyces gilkeyae singer & a.h. sm. mem. torrey bot. club 21: 21 (1960). = martellia gilkeyae (zeller & c.w. dodge) singer & a.h. sm., mem. torrey bot. club 21: 32 (1960). fig.1. basidiomata of arcangeliella scissilis. fig. 2. spore of arcangeliella scissilis seen under the sem. arcangeliella scissilis, a rare american semihypogeous fungus 23 basidioma globose to tuberiform, 23–35 mm diameter in fresh. peridium smooth, cream with orange, to dark reddish tones. in wet weather the surfase changes to viscid or muscilaginose. the consistency of basidioma is solid, sofl in fresh conditions and coriaceus when dry, with venous surface. gleba partially exposed at base of basidioma, where a rudimentary prolonged stipe-like formation can be observed (fig. 1). gleba in section appears with a labyrinthic structure, with chambers 0.1–0.4 mm diameter, full of spores, showing a cream colour when mature. columella absent. sections of the fresh basidioma exuding a white latex, which changes to lemon yellow in a few minutes. smell and taste indistinct. when observed under the light microscope it is possible to distinguish an epicutis made up of filiform hyphae, 8–12 × 5–7 μm, with a gelified apical cell. peridium filamentous, 120–160 μm thick. the external layer shows a yellowish orange pigmentation, made up with modified hyphae, 2–3 μm diameter. hymenial context made up with amber colour laticiferous hyphae 10 μm diameter, without septa, and hyaline 4-6 μm diameter hyphae, septate, without clamp connections, but with some rare sphaerocyst nests at the context centre. sphaerocysts, 8–14 μm diameter. subhyme­ nium made up with globose elements, 9–23 μm diameter, rare. basidia cylindrical to elliptical, 45–55 × 9–11 μm, hyaline which does not change in the presence of koh, with 1–2–4 sterigmata, 5–6 μm long. cystidia claviform, 40–60 × 10–12 μm, hyaline. spores globose, yellowish in mass, 10–16 μm diameter, including spines, which are cylindrical to filiform 2–4 μm long, amyloid (fig. 2). discussion after a first look to the studied material, we thought that it was an arcangeliella, due to the presence of an abundant white latex, rudimentary to absent stipe and a partially nude gleba at the base. thanks to the excellent article published by thiers (1984), on the genus arcangeliella, it has been possible to include our material within this genus. the material from costa rica was coincident with that of zelleromyces gilkeyae (singer, smith 1960) that is why we decided to identify both taxa. nevertheless, the problem of the genera delimitation in the astrogastraceous is a big one, according to singer and a.h. smith (1960). the production of an abundant latex in fresh is a fundamental character in zelleromyces, which makes possible the separation with gymnomyces and martellia, without latex. however, this character is difficult to observe when working with dry material or herbarium specimens. thus, beaton et al. (1984) and zhang and yu (1990) have proposed, as a better character, the spore ornamentation. in this way, they suggest to include within the genus zelleromyces only species with reticulate, subreticulate or winged spores. in martellia those species with warty to spiny spores and in gymnomyces all kind of spores; being possible the separation between the last two groups by the presence or absence of sphaerocyst nests, which are not always easy to locate. thus, none of these methods is really functional. 24 f. d. calonge and m. mata in previous works (moreno-arroyo et al. 1998a, b, 1999; calonge, vidal 1999, 2001; calonge, martín 2000; vidal et al. 2002), we have followed beaton´s et al. concept (1984), but in the actual situation the presence of abundant latex and spiny spores places our material in an intermediate position, among zelleromyces, gym­ nomyces and cystangium. within this chaos, vidal (2004) has been trying to get a reasonable explanation rediscribing the genus arcangeliella (cavara 1900) to which the genus zelleromyces is synonymysed. thus, within the actual vidal´s concept, all the genera from the astrogastraceous, lacking stipe or any rest of it, from the sterile base or columella, are included in arcangeliella. this is the case of our material from costa rica. other important characters, besides latex presence, are hymenial trama, generally homomerous; despite the possibility to observe some dispersed sphaerocysts, in a. scissilis. on the other hand the spores may be reticulate, subreticulate, warty, spiny, globose to ellipsoid. at the same time, the other species with well-developed stipe and percurrent columella are included in a restored genus, gastrolactarius r. heim ex vidal (vidal 2004). regarding to molecular biology studies, carried out on this group, miller et al. (2001) have demonstrated that arcangeliella and zelleromyces aligne within lactar­ ius section, while macowanites, gymnomyces, cystangium and martellia do within russula clades. concerning the ecology of arcangeliella scissilis, as far as we know, it grows under quercus sp.; other authors have collected it under acer (singer, smith 1960), alnus, picea, tsuga (desjardin 2003) and in the case of the so named martellia gilkeyae it was found under corylus (singer, smith 1960). its geographical distribution, until now, was limited to west of usa (desjardin 2003), thus, the costa rican collection should be the first record outside the united states. conclusions arcangeliella scissilis represents a rare taxon which is limited to western united states and costa rica. it seems to be associate to various species of vascular plants, such as acer, alnus, quercus, picea, tsuga and corylus. from the phylogenetic point of view, arcangeliella fits well within the lactarius clades. acknowledegements. we wish to express our sincere gratitude to mr. isaac lópez, as a parataxonomist from the inbio, for his valuable collaboration, collecting interesting mycological samples for scientific study. to mr. juan carlos hernández, for his technical assistance during the preparation of the manuscript. an anonymous reviewers are thanked for valuable comments and suggestions to this manuscript. arcangeliella scissilis, a rare american semihypogeous fungus 25 references beaton g., pegler d.n., young t.w.k.1984. gasteroid basidiomycota of victoria state, australia. 2. rus­ sulales. kew bull. 39: 669-698. calonge f.d., martín m.p. 2000. morphological and molecular data on the taxonomy of gymnomyces, martellia and zelleromyces (russulales). mycotaxon 76: 9-15. calonge f.d., mata m. 2005. crucibulum laeve var. magnum var. nov. y arcangeliella scissilis, encontrados en costa rica. bol. soc. micol. madrid 29: 43-48. calonge f.d., vidal j.m. 1999. gymnomyces ammophilus vidal & calonge sp. nov., encontrado en portugal. bol. soc. micol. madrid 24: 65-70. calonge f.d., vidal j.m. 2001. macowanites vinaceodorus sp. nov. 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(russulales), a gasteroid fungus from the south of spain. cryptogamie, mycol. 19: 107-111. moreno-arroyo b., gómez j., calonge f.d. 1998b. zelleromyces meridionalis (russulales, elasmomyce­ taceae) a new species from spain. mycotaxon 69: 467-471. moreno-arroyo b., gómez j., calonge f.d. 1999. gymnomyces dominguezii sp. nov. from spain. mycol. res. 103: 215-218. singer r., smith a.h. 1960. studies on secotiaceous fungi. ix. the astrogastraceous series. mem. torrey bot. club 21: 1-112. thiers h.d. 1984. the genus arcangeliella cav., in the western united states. sydowia 37: 296-308. vidal j.m., calonge f.d., martín m.p. 2002. macowanites ammophilus (russulales) a new combination based on new evidence. rev. catalana micol. 24: 69-74. vidal j.m. 2004. arcangeliella borziana and a. stephensii, two gasteroid fungi often mistaken. a taxonomic revision of lactarius­related sequestrate fungi. rev. catalana micol. 26: 59-82. zhang b., yu y.n. 1990. two new species of gasteroid russulales from china, with notes on taxonomy of gymnomyces, martellia and zelleromyces. mycol. res. 94: 457-462. 2013-06-22t22:41:01+0100 polish botanical society parasites of zooplankton and periphyton assemblages in the littoral zone of lakes in drawa national park, poland maria wolska and kinga mazurkiewicz-zapałowicz west pomeranian university of technology in szczecin, department of hydrobiology ichthyology and biotechnology of reproduction, 4 k. królewicza st., pl-71-550 szczecin wolska m., mazurkiewicz-zapałowicz k.: parasites of zooplankton and periphyton assemblages in the littoral zone of lakes in drawa national park, poland. acta mycol. 48 (1): 51–59, 2013. studies on parasitism in zooplankton and periphyton assemblages were carried out on samples of water collected from lakes płociczno and płociowe in the drawa national park in 2008-2011. occurrence of the fungal mycobiotal and protozoan parasites of invertebrates was recorded in both lakes. parasitism of chydorus sp. (cladocera) by saprolegnia sp. occurred on 0.8% of individuals of the host population, of brachionus calyciflorus (rotifera) by microsporidium sp. on 1.2% of the host population, and of nematoda members by pythium sp. on 5% of the host population. parasites were recorded only in springtime. key words: mycobiota, microsporidia, microparasities, plankton, periphyton introduction plankton and periphyton are a complex mixture of organisms that make a significant contribution to the littoral zone of lakes. the composition of these assemblages is affected by abiotic and biotic factors and is under continuous and dynamic change. the periphyton consists of a network of complex configurations of organisms with complex interspecies relationships. in the aquatic environment, the least known are still the relationships between the invertebrata (kingdom: animalia) and fungi (kingdom: fungi/mycota), as well as between invertebrata and protozoan fungal analogues (kingdom: protozoa) and chromistan fungal analogues (kingdom: chromista). parasitism is particularly important in these relationships. it often determines the density of individual species in water. organisms that comprise the plankton and periphyton are potential hosts of various ectoand endoparasites. there are susceptible in varying degrees to infection, the success of which depends also on genetic factors (stirnadel, ebert 1997). parasites of plankton and periphyton organisms additionally modify the physiological and behavioral processes of their hosts, which is followed by changes in density, survival, fecundity and lifespan of the host acta mycologica vol. 48 (1): 51–59 2013 doi: 10.5586/am.2013.007 52 m. wolska and k. mazurkiewicz-zapałowicz (mirakle 1977; ruttner-kolisko 1977) and growth rate of the population (thomas et al. 2011). there has been continued interest in zooparasites present in the aquatic habitat for the last 150 years, since the first zooparasitic protozoan fungal analogues species, amoebidium parasiticum cienk. was recorded on the copepod cyclops (cyclopidae) (whisler 1962; whisler, fuller 1968). although it is known that plankton and periphyton assemblages include fungi, protozoan and chromistan fungal analogues saprotrophs, parasites and predators (kiziewicz and czeczuga 2003), our knowledge of their occurrence, as well as their contribution and significance in sustaining the equilibrium of aquatic ecosystems, is inadequate. studies using various approaches aimed at recognition of parasites of plankton and periphyton organisms are therefore necessary. they will provide new information on the functioning and relationships in assemblages of parasites and their hosts. such knowledge is of particular value in the case of natural bodies of water, particularly those located in environmentally sensitive and protected areas. the proper functioning of successive links of the microbial loop (including plankton and periphyton parasites) in these ecosystems is of particular significance. the aim of this study was to determine the occurrence of parasites in zooplankton and periphyton assemblages in the littoral zone of two lakes in drawa national park in poland. materials and methods studies on parasitism in the zooplankton and periphyton assemblages were carried out on samples of water collected from two lakes in the drawa national park. samples were collected from lake płociowe, which is mesotrophic and has no throughflow, in 2008-2009, and from lake płociczno, which is eutrophic and has throughflow, in 2011. samples were collected three times each year, in april/may (spring circulation), august (summer stagnation) and november/december (autumn circulation), from five sites in the littoral zone of each lake. zooplankton samples were taken with a 5-l “toń” sampler from water 0-1 m deep. on each occasion, 50 l of water was collected in 10 hauls. the water was filtered through a plankton net with a 44 μm diameter mesh. the total sample of zooplankton from the two lakes resulted from filtration of 2250 l of water. additional zooplankton assemblages were collected using a net for a 2 min period through the water among plants. the samples of zooplankton were then preserved in 4% formaldehyde solution. rotifera and crustacea (invertebrata) were counted and identified to species in a 1ml chamber using a microscope (nikon eclipse e 200) at 60-240x magnification. taxonomy was determined by morphology according to flössner (1972), kiefer and fryer (1978), and radwan et al. (2004). periphyton samples were collected from submerged shoots of the hydrophytes, phragmites australis (cav.) trin. ex steud. and carex acutiformis l. two pieces, approximately 10 cm long, were cut from each shoot and preserved in 5% formaldehyde solution. a total of 180 pieces from 90 shoots was analysed from each lake. two methods were used to determine the occurrence of parasites on the periphyton organisms. the first was microscopic observation of a suspension collected from parasites of zooplankton and periphyton in the littoral 53 approximately 3600 cm2 of plant surface. the second was similar to that used in the study of zooplankton. zooplankton and periphyton organisms were assessed for their state of health and possible interactions with parasites, mainly with those from protozoan and chromistan fungal analogues. the parasites were determined based on morphology according to skirgiełło (1954) and khulbe (2001) using a nikon eclipse e 200 biological microscope and 60-800x magnification. nomenclature of fungi, protozoan and chromistan fungal analogues was adopted according to index fungorum (www. indexfungorum.org) and their classification according to kirk et al. (2008). results and discussion the zooplankton in the littoral zone of lake płociowe was dominated by cladocera (92% of the total population) and of lake płociczno by rotifera (50% of the total). the mean abundance of the zooplankton organisms was 213 individuals l -1 in lake płociowe in 2008-2009, and 140 individuals l -1 in lake płociczno in 2011. the mean abundance of particular taxonomic groups of zooplankton in each lake in each season in 2008-2011 is shown in figure 1. host responses to parasites in zooplankton assemblages were assessed for 34, 25 and 7 taxa of animals: rotifera (phylum), cladocera (phylum arthropoda) and copepoda (phylum arthropoda), respectively, in lake płociowe, and for 57, 30 and 18 taxa of rotifera, cladocera and copepoda in lake płociczno. protozoan and chromistan fungal analogues parasites of the zooplankton organisms were found to occur only in spring, in 2009 in lake płociowe and in 2011 in lake płociczno. the possible explanation of such short-duration, seasonal occurrence of parasites is a highly dynamic succession, which results in elimination of protozoan and chromistan fungal analogues by bacteria. bacteria are the most fig. 1. the mean abundance of particular taxa of zooplankton in lakes płociowe and płociczno in 2008-2011; sp – spring, su – summer, a – autumn. 54 m. wolska and k. mazurkiewicz-zapałowicz commonly recorded destructive agents, being continuously present in large numbers and having high biochemical activity (ebert 1995; decaestecker et al. 2005). only one case of parasitism of chydorus sphaericus (cladocera, chydoridae) (o.f. müller) by saprolegnia sp. (chromistan fungal analogues, kingdom chromista) was recorded in lake płociowe (fig. 2). infection was observed on 0.8% of fig. 2. chydorus sphaericus (cladocera) infected with saprolegnia sp. – zoosporangia with zoospores. fig 3. brachionus calyciflorus (rotifera) infected with microsporidium sp. parasites of zooplankton and periphyton in the littoral 55 the chydorus population, which accounted for 6.7% of all cladocera. there were no other fungi and fungal analogues parasites of zooplankton in lake płociowe. an example of parasitism of brachionus calyciflorus (rotifera) by protozoan microsporidium sp. was recorded in lake płociczno (fig. 3). the parasite was recorded on 1.2% of the b. calyciflorus population. these single examples of parasitism, although not very impressive, may be considered representative and credible because they were recorded in very large samples of zooplankton. comparision of the results with similar studies. as already mentioned, the first zoopathogenic species observed in aquatic environment was protozoan fungal analogue amoebidium parasiticum cienk. parasitising on cyclopoidae (copepoda) (whisler 1962). parasites coelomomyces psorophorae couch ex couch (kingdom: fungi, phylum: chytridiomycota) were identified by guilvard et al. (1977) and goettel (1987) on mosquito larvae. the larvae of these insects were also infected by two species of protozoan fungal analogues, i.e. hyalinocysta chapmani e.i. hazard & oldacre (andreadis 2002) and amblyospora sp. (nasci et al. 1987). in poland, the occurrence of fungi and fungal analogues on planktonic arthropods (cladocera, copepoda and rotifera) was studied by czeczuga et al. (2000) and kiziewicz (2004). in these studies a total of 29 cladocera species were identified, among which dominant were chromistans lagenidium gigantem couch and ancylistes cladocerarum fritsch (fungi, phyllum: zygomycota). they were observed throughout the year in all phases of the development of these crustaceans. species l. giganteum was also most frequently identified on rotifera, together with olpidium granulatum karling (kingdom: fungi, phyllum: chytridiomycota). mycoses of rotifers have also been caused by other species, i.e. chytriomyces aureus karling (kingdom: fungi; phyllum: chytridiomycota) and chromistan myzocytium microsporum (karling) sparrow and m. zoophthorum sparrow. most published studies concern the occurrence of parasitic microorganisms on dead (czeczuga et al. 2000; godlewska et al. 2003) or alternatively living zoohydrobionts, though in laboratory conditions (lass, ebert 2006; wolinska et al. 2009). to date, however, not many in situ studies related to parasitic infections of live zooplankton and zooperiphyton. most often they concerned cladocera parasites, including daphnia spp., forming at times large populations in small water reservoirs (brambilla 1983; stirnadel, ebert 1997; wolinska et al. 2004). there are also studies on rotifera parasites occurring in rivers (gorbunov, kosova 2001). results presented here are similar to and comparable with those reported by goren and ben-ami (2013), who studied the ecological relationships between cladocerans and their endoparasites in 204 freshwater bodies. they recorded a total of 21 taxa of endoparasites on 14 taxa of cladocera. only two parasites belonged to kingdom fungi, i.e. metschnikowia bicuspidate (metschn.) (phyllum: ascomycota) and brood pouch parasite. four of them were bacteria species, eleven constituted protozoan organisms from microsporidium group, while four were unclassifified parasites. the most common endoparasites were bacteria – spirobacillus cienkowskii metchnikoff and protozoan fungal analogeus – glugoides intestinalis chatton. comparatively few species of parasitic eukaryotes was reported by gorbunov and kosova (2001), who found only one representative of the chromistan fungal analogues (pythium sp.) and two species of fungi, (catenaria anguillulae sorokin and olpidium 56 m. wolska and k. mazurkiewicz-zapałowicz gregarium nowakowski) in more than 20 species of rotifera from water bodies in delta of the volga river. pythium sp. was, however, the most commonly encountered and present in water throughout the year. other authors have reported, however, a greater contribution of fungi and fungal analogues in the parasitism of zooplankton. czeczuga et al. (2000) isolated 29 species of parasites from dead specimens of cladocera. the most common species, recorded throughout the year, in all growth stages of these small crustaceans were the chromistan fungal analogues lagenidium giganteum couch and fungal ancylistes sp. (phyllum: zygomycota). the former, together with olpidium granulatum (kingdom: fungi; phyllum: chytridiomycota), was also the most common on rotifers. mycoses of rotifera were caused also by other fungi, namely chytriomyces aureus karling and chromistan fungal analogues: myzocytiopsis microspora (karling) m.w. dick [= myzocytium microsporum] and myzocytiopsis zoophthora (sparrow) m.w. dick [= myzocytium zoophythorum]. wolinska et al. (2009) recorded four species of protosoan fungal analogues (microsporidium group), one species of fungi and nine chromistan fungal analogues members among 14 parasites infecting daphnia. czeczuga et al. (2000) found, in studies of parasitism of copepoda, that the occurrence of a particular fungal and protozoan and chromistan fungal species was limited to a single season. in this study also the tendency for seasonal occurrence of mycobiotal zooparasites was indicated. their abundance increased in spring, resulting from the local and seasonal presence and abundance of hosts. therefore, where the host occurs only locally and seasonally, parasitism will be local and short-term, with low-moderate significance. reasons for dynamic occurrence of parasites of zooplankton and periphyton. the sporadic occurrence of parasites may result from the generally low abundance of zooplankton. this was observed in both lakes (fig. 1). in lake płociczno, the low incidence of infestation of brachionus calyciflorus (rotifera) by microsporidium sp. was associated with the low abundance of the potential host in the zooplankton (although the abundance of zooplankton in general was high). this observation agrees with duffy et al. (2005), who pointed out that low abundance of the host population limits the spread of its parasite. infestation of b. calyciflorus by microsporidium asperospora may, however, be more common (even on 30-40% of the host population), as observed in water bodies of the volga delta (gorbunov, kosova 2001). the frequency of occurrence of parasites on zooplankton organisms may be affected additionally by the size and shape of the host organism. brambilla (1983), and stirnadel and ebert (1997) found a positive correlation between size of the host (daphnia species) and percentage of population infested. populations of d. magna straus (>2.4 mm long), d. pulex leydig (>1.7 mm long) and d. longispina müller (<1.3 mm long) were infested, on average, at 85%, 53.6% and 38.6%, respectively. the tendency to colonize and infest larger organisms results not only from exposure of a greater surface area of the host to the parasite but is also associated with the host’s physiology. larger species of daphnia filter more water while feeding, and thereby absorb more spores of mycobiota and invasive stages of parasites present in the water (ebert 1995). this suggestion seems to be indirectly confirmed by our own studies, as the largest species of daphnia, i.e. d. magna was not found in littoral plankton. this species is most susceptible to infection by microorganisms and they parasites of zooplankton and periphyton in the littoral 57 increase epidemic spread of pathogens in the population. this could, however, have resulted from the early infestation of d. magna and its consecutive elimination. infected and diseased individuals are more distinctive and can be the preferred target of planktivorous fish hunting by sight (yan and larsson 1988). reduction of the host by planktivorous fish may lead to decreased parasites’ diversity (duffy et al. 2005; wolinska et al. 2009). success of infection of zooplankton by both ectoand endoparasites results also from the susceptibility of the host or pathogenicity of the parasite. variation in susceptibility among hosts may be affected by the structure and composition of the cuticle. variation in pathogenicity of the parasite may depend also on its biochemical activity. endomycosis of chydorus sphaericus caused by saprolegnia sp. (fig. 2) may result from proteolitic or cytotoxic activity of the latter. both kinds of activity have been observed in s. monoica pringsh., and s. parasitica coker (mazurkiewicz-zapałowicz et al. 2008). they may cause degeneration of the host’s hemocoel (tissue spaces) resulting in death of the host. in parasitism of brachionus individuals by microsporidium group, eventual death of the host is associated with the accumulation of spores (fig. 3). greater incidence of parasitism seems to result also from the horizontal distribution and migration of zooplankton in the littoral zone, which increases the chance of meeting the spores of parasites. this suggestion is supported by studies of ebert (1995), who found, in laboratory experiments, a positive correlation between accumulation of parasite spores and the level of parasitism of zooplankton. in the periphyton assemblages, consisting of protozoa (protozoan phylum ciliophora: coleps sp., peritricha and ameboid protists: testacea), rotifera, ostracoda, cyclopoida (nauplii), cladocera (chydoridae – alona guttata sars., a. guadrangula (o.f.m.), chydorus sp.), oligochaeta (chaetogaster von baer, stylaria lamarck), nematoda and diptera (larval stages of chironomidae), and nematoda. only nematoda, in lake płociczno, were parasitized by pythium sp. this parasite appeared only in spring 2011 and reached a prevalence of 5%, which is not high considering that nematoda accounted for 11% of organisms in the periphyton. while most of the species of pythium are pathogenic to plants recent studies on daphnia (cladocera) in europe (wolinska et al. 2009), and on asplanchna gosse (rotifera) in the usa (thomas et al. 2011) and russia (gorbunov, kosova 2001) have showen that the genus also includes species pathogenic to water invertebrates. conclusions this study confirms the presence of diverse, unique and specific interactions resulting from the presence of parasites within freshwater zooplankton and periphyton assemblages. the diversity of these interactions may result from a high level of adaptability of the parasites to the local habitat. this means that recognition of dynamic relationships in the microbial loop system is difficult, which emphasizes the need for further studies. acknowledgements. the studies were supported by ministry of science and higher education, grant no. nn 304 385639. the authors of the paper thank the management of drawa national park (poland) for their help in collecting the research material. the authors are grateful to the anonymous reviewers for valuable comments and suggestions on the manuscript. 58 m. wolska and k. mazurkiewicz-zapałowicz references andreadis t.g. 2002. epizootiology of hyalinocysta chapmani (microsporidia: thelohaniidae) infections in field population of culiseta melanura (diptera:culicidae) and orthocyclops modestus (copepoda: cyclopidae): a tree-year investigations. j. invert. pathol. 81 (2): 114-121. http://dx.doi.org/10.1016%2fs0022-2011%2802%2900154-4 brambilla d.j. 1983. microsporidiosis in a daphnia pulex population. hydrobiologia 99: 175-188. http://dx.doi.org/10.1007%2fbf00008769 czeczuga b., godlewska a., kozłowska m. 2000. zoosporic fungi growing on the carapaces of dead zooplankton organisms. limnologica 30: 37-43. http://dx.doi.org/10.1016%2fs0075-9511%2800%2980040-7 decaestecker e., declerck s., de meester l., ebert d. 2005. ecological implications of parasites in natural daphnia populations. oecologia 144 (3): 382-390. http://dx.doi.org/10.1007%2fs00442-005-0083-7 duffy m., hall s.r., tessier a. j., huebner m. 2005. selective predators and their parasitized prey: are epidemics in zooplankton under top-down control? 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(erratum in j. am. mosq. control. assoc. 1990 mar. 6 (1): 151). radwan s., bielańska-grajner i., ejsmont-karabin j. 2004. rotifers (rotifera), freshwater fauna of poland. 32. oficyna wydawnicza tercja, łódź (in polish). parasites of zooplankton and periphyton in the littoral 59 ruttner-kolisko a. 1977. the effect of the microsporid plistophora asperospora on conochilus unicornis in lunzer untersee (lus). arch. hydrobiol. beih. ergebn. limnol. 8: 135-137. skirgiełło a. 1954. grzyby niższe. pragrzyby i glonowce. przewodnik morfologiczno-systematyczny z klu-kluczami do oznaczania. pwn, warszawa (in polish). stirnadel h.a., ebert d. 1997. prevalence, host specificity and impact on host fecundity of microparasites and epibionts in three sympatric daphnia species. j. anim. ecol. 66: 212-222. http://dx.doi.org/10.2307%2f6023 thomas s.h., housley j.m., reynold a.n., penczykowski r.m., kenline k.h., hardegree n., schmidt s., duffy m.a. 2011. the ecology and phylogeny of oomycete infections in asplanchna rotifers. freshwater biol. 56: 384-394. http://dx.doi.org/10.1111%2fj.1365-2427.2010.02505.x whisler h. c. 1962. culture and nutrition of amoebidium parasiticum. am. j. bot. 49 (3): 97-191. whisler h.c., fuller m.s. 1968. preliminary observations on the holdfast of amoebidium parasiticum. mycologia 60 (5): 1068-1079. http://dx.doi.org/10.2307%2f3757291 wolinska j., keller b., bittner k., lass s., spaak p. 2004. do parasites lower daphnia hybrid fitness? limnol. oceanogr. 49 (2): 1401-1407. http://dx.doi.org/10.4319%2flo.2004.49.4_part_2.1401 wolinska j., giessler s., koerner h., 2009. molecular identification and hidden diversity of novel daphnia parasites from european lakes. appl. environ. microbiol. 75 (22): 7051-7059. http://dx.doi.org/10.1128%2faem.01306-09 yan n.d., larsson j.i.r. 1988. prevalence and inferred effects of microsporidia of holopedium gibberum (crustacea: cladocera) in a canadian shield lake. j. plankton res. 10: 875-886. http://dx.doi.org/10.1093%2fplankt%2f10.5.875 index fungorum (www.indexfungorum.org) pasożyty związane z organizmami planktonowymi i peryfitonowymi strefy litoralowej jezior drawieńskiego parku narodowego streszczenie w latach 2008-2011 badano występowanie mikroorganizmów pasożytniczych na gatunkach tworzących zooplankton i peryfiton litoralowy w jeziorach płociczno i płociowe w drawieńskim parku narodowym. występowanie patogenów na organizmach tych zespołów stwierdzono jedynie wiosną. udokumentowano porażenie chydorus leach (cladocera) przez saprolegnia sp., brachionus calyciflorus pall. (rotifera) przez microsporidium sp. oraz przedstawiciela nematoda przez pythium sp. porażenie przez patogeny dotyczyło 0,8% populacji chydorus sp., 1,2% populacji brachionus calyciflorus oraz 5% populacji niezidentyfikowanych do gatunku nicieni (nematoda). w pracy przeanalizowano uzyskane wyniki na tle innych, podobnych badań i przedyskutowano prawdopodobne przyczyny niewielkiego stopnia porażenia organizmów planktonowych i peryfitonowych przez pasożyty. wskazano także na obserwowaną przez niektórych autorów zależność występowania pasożytów od dynamiki sezonowej występowania ich żywicieli. 2013-06-22t22:38:16+0100 polish botanical society the occurrence of, and economic losses caused by armillaria in the western carpathian mts 1adam kaliszewski, 2paweł lech and 2tomasz oszako 1department of forest economics, forest research institute sękocin stary, braci leśnej 3, pl-05-090 raszyn, a.kaliszewski@ibles.waw.pl 2department of forest phytopathology, forest research institute sękocin stary, braci leśnej 3, pl-05-090 raszyn k a l i s z e w s k i a., l e c h p., o s z a k o t.: the occurrence of, and economic losses caused by armillaria in the western carpathian mts. acta mycol. 42 (2): 219-233, 2007. an investigation carried out in the western carpathian mountains (ujsoły, węgierska górka, ustroń and wisła forest districts) demonstrated a strong relationship between dieback in norway spruce stands and the intensity of occurrence of armillaria ostoyae. for the most endangered site types – mountain deciduous forest (lg) and mountain mixed forest (lmg), analyses of losses of annual volume increment and of stand productivity were performed, and their financial dimensions determined. the greatest losses – of about 8 m3/ha/year for tree stands of the age of 100 years, and 400 m3/ha for the rotation period – were found for lg (mountain broadleaved forest) site type. key words: armillaria occurrence, economic losses, spruce root rot, carpathian mountains introduction root rots caused by fungi of the genus armillaria represent one of the most important problems for polish forestry. the most serious losses due to them are to be noted in coniferous stands (s i e r o t a et al. 2003). according to data from the state forests administration, damage due to armillaria extended over a total of 144,000 ha in 1999, cf. more than 200,000 ha in 2003. five species of the genus armillaria have been recorded in poland, namely a. borealis, a. gallica, a. ostoyae, a. cepistipes and a. mellea (ż ó ł c i a k 1991, 1999a). while a. ostoyae, a. cepistipes and a. gallica are present across the country, a. borealis is a species of northern and central parts, and a. mellea is confined to just a small area near gubin by the german border (ż ó ł c i a k 2003). a. ostoyae has been reported from coniferous, broadleaved and mixed stands, and in a wide range of forest habitats from fresh coniferous forest, through to moist broadleaved forest, alder-ash woodland and fertile broadleaved forest habitat in acta mycologica vol. 42 (2): 219-233 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 220 a. kaliszewski et al. the mountains (lmg, lg, lłg) – including to an altitude of some 1100 m a.s.l. (ż ó ł c i a k 1999b). however, its preference is for coniferous species, especially spruce, scots pine and fir. in poland a. ostoyae displays a decided dominance in managed forests. it has been noted on the greatest variety of shrubs and trees (23 species), including all the tree species native to poland that form forests, i.e. scots pine, norway spruce, fir, oaks, beech and birch (ż ó ł c i a k 2003). in the pine stands of the polish lowlands, and most notably in the spruce forests of the western carpathians, armillaria root rot is present as an epiphytosis (c a p e c k i 1994, 1997; l e c h 2003). furthermore, recent years have seen it appear in stands that had been considered resistant – in the beech forests of the bieszczady mountains and pomerania, as well as on oaks throughout the country (ż ó ł c i a k 1999b; ł a k o m y , s i w e c k i 2000). it may thus serve a useful indicator of the state of health of a forest, inter alia on account of the constancy of colonisation of the substratum (tree boles and roots), as well as the length of the disease process. its presence depicts areas with a greater predisposition to other stress and injurious factors, being an indicator of the pathological state of the stand, as well as pointing to changes of an ecosystemic nature. it not only does economic damage, but also brings about changes in the functioning of a biocoenosis, leading to degradation of stands, a reduction in the productivity of the habitat, and even deforestation. this impact has been registered in the spiral disease model (m a n i o n 1981), wherein it features as a predisposing factor, an initiating factor and a factor co-participating in the treedieback process. research has not so far been carried out in poland with a view to discovering the level of the damage sustained by forestry through the reduced productivity of stands colonized by pathogenic fungi. as a consequence of the harmful impact of fungi from the economic point of view, damage is done to single trees and to whole groups of them, a possible result being the dieoff of whole patches of forest, necessitating premature felling. the consequent loss of growing stock may play a major role in limiting the productive potential of forest habitats. the means of calculating losses due to damage arising in forests are as set out in the agricultural and forest land protection act 1995 (ustawa 1995). the level of one-off compensation for the premature felling of a stand is set as the difference between the expected value of the stand at rotation age, as detailed in the forest management plan, simplified forest management plan or forest inventory, and the value at the moment of felling (p o d g ó r s k i et al. 2001). the principles for the valuation of stands and level of damage in the case of their premature felling are in turn laid down in the 2002 regulation of the minister of the environment on one-off damages for the premature felling of a stand (rozporządzenie 2002). the work presented here had as its aim the determination of the environmental conditioning behind the occurrence of armillaria root rot in stands of the western carpathians. in addition, the work sought to determine the size of losses as expressed in terms of reduced stand volume increment, and the current value of lost timber expressed per hectare and per year. in this, no account was taken of the losses resulting from the impact on growth of other stress factors like insect pests, air pollution and weather anomalies (droughts). within poland, the western part of the carpathians encompasses the chain of mountains comprising the beskid śląski, żywiecki, mały, wyspowy and sądecki ranges, as well as the gorce and tatra mountains. the armillaria in the western carpathian mts 221 forests in this region are dominated by spruce, whose average volume share in the stand structure at present exceeds 62% (s z a b l a 2003), locally even attaining 95% (c a p e c k i 1994). the shares taken by other species are thus markedly lower – on average beech accounts for 19% of volume, scots pine and fir for 6% each, oak for 3%, birch for 2% and other species also for 2%. average annual increment in these stands exceeds 4.5 m3/ha (s z a b l a 2003). forests in this western part of the carpathians are subject to the impact of many biotic, abiotic and anthropogenic stress factors, including primary and secondary insect pests (most especially bark beetles), infectious fungal diseases (first and foremost those giving rise to root rot), weather anomalies (drought, wind and snow), and air pollution (z w o l i ń s k i 2003). root pathogens are of particular significance in this area – fungi of the genus armillaria (mainly a. ostoyae), and heterobasidion annosum, which occurs more frequently here than anywhere else in poland’s forests (l e c h 2003). material and methods the area chosen for study was the ujsoła forest district, located in the boundary zone between the beskid żywiecki and beskid śląski ranges and typical of the region in terms of its stand structure and the threats posed to it. it is characterised by a high (and in recent years increasing) level of damage posed to stands by armillaria root rot, as well as by differences in the degree of damage from place to place. this fact made possible the establishment in 2001 and 2002, within the district (and specifically its ujsoła sub-district), of a total of 26 2-are observation plots so selected as to take account of both the differences in the level of threat and different stand and habitat parameters (altitude above sea level and the stand structure in terms of age and species). the plots were arranged in groups of between 3 and 10 along 4 transects 700 m to 4 km long. these were found in uniform spruce forest and mixed stands (spruce-beech or spruce-beech-fir), in high-mountain coniferous forest habitat (bwg), mountain mixed coniferous forest (bmg), mountain mixed broadleaved forest (lmg) and mountain broadleaved forest (lg), at altitudes of between c. 600 and c. 1350 m a.s.l. observation plots were subjected to assessments of the occurrence of armillaria, including that on dead stumps as well on trees. the results of this assessment were compiled, together with information on the volume of deadwood removed in the years 1987-2001, by reference to stand areas in which observation plots were located (use was made of volume expressed per year and per hectare). they were then the subject of analysis of variance taking account of forest type and stand species structure as sources of variability. also the potential economic losses incurred as a result of the lowering of stand values on account of root-rot attacks were assessed. estimation of damage was done for spruce stands, i.e. those most threatened by armillaria and also prevalent in the western carpathians, occurring on the two habitat types lg and lmg. the small number of plots in the bmg and bwg habitats did not allow for any analysis in these forest habitats. calculations also made use of data on the amount of deadwood generated in the years 1985-2001. in determining the value of the damage, use was made of the formula below [1], this being a modification of that proposed in the regulation of the 222 a. kaliszewski et al. minister of the environment on one-off compensation for premature felling of a stand (rozporządzenie 2002). this formula serves in detailing losses due to the reduced volume increment resulting from the partial damage to the stand (z a j ą c et al. 1998). the formula is as follows: s = (wu – wi) × (zi – zs) × p [1] where: s is the loss due to the reduction in increment caused by the partial damaging of the stand; wu is the expected value of 1ha of standing trees in a stand, in line with the expenditure necessary for its development to age u; wi is the expected value of 1 ha of standing trees in a stand in line with the expenditure necessary for its development to age i;u is the rotation age of the stand subject to estimation; zi is the stock density prior to the damage; i is the current age of the stand; zs is the expected stock density of the stand following the thinning out of damaged trees, p is the area of the stand in ha. the index of expected stand value allows for a determination of the value of a stand during each year of its existence (from establishment of the plantation to felling at the rotation age). the method rests on the assumption that the whole period of life of the stand includes only two times at which direct estimates of its value can be made, i.e. 1) the age of establishment of the plantation (i=1), at which time the value equals the cost of establishment, and 2) the rotation age u (i=u), at which time the value is given by the combined value of the harvested wood assortments. the value of a stand at age i (wi) is calculated by reducing its value at the rotation age using a coefficient that differs in line with stand age (p a r t y k a , p a r z u c h o w s k a 1993; p a r z u c h o w s k a et al. 1997; z a j ą c et al. 1998). indices of expected values of stands at ages i and u (with account being taken of species and stand quality classes) are set out in the tables of stand value indices constituting an annex to the aforementioned regulation. these tables contain values in conversion units, i.e. ones expressed in m3 of raw timber. the values obtained from the valuation, expressed in terms of conversion units, are multiplied by the average wood selling price published annually by the central statistical office for the purposes of calculation of the forest tax. the assessment of the expected (potential) value of losses is made in three stages: 1. determination of the potential loss in the capacity to generate stand increment, 2. estimation of the potential decline in stock density of stands colonized by armillaria, 3. estimation of the potential economic losses in the stand as armillaria-induced disease progresses. the starting point for completion of the first stage of the research was provided by data on the mean annual volume removed in the period 1985-1997 through the clearing of deadwood from variously-aged spruce stands or stands with a considerable share of spruce. information on stands in the lg habitat was available for 16 research plots taking in stands aged 6 to 100 years. in the case of stands of the lmg habitat, the information came from 5 plots established in stands of ages 75 to 125 (tab. 1). 224 a. kaliszewski et al. the functional relationships obtained made it possible to determine potential losses in terms of stand increment. the potential increment of spruce stands was established on the basis of the tables of stand yield and increment (s z y m k i e w i c z 2001), accepting that the further calculations would make use of data for current increment of wood with diameter of at least 7 cm overbark, with a stock density equal to 1.0. it was assumed that stands of habitat lg were of quality class i, while those of habitat lmg were of class ii (tr a m p l e r 1990). losses to stand increment were defined as the difference between their potential current increment overbark and the volume of deadwood removed. the results were presented in absolute terms (in m3/ha/year), as well as relatively – by reference to the percentage reduction in potential increment (tabs 2 and 3). the second stage involved determination of the scale of the reduction in the potential stock density of those stands subject to the unfavourable impact of armillaria fungi. to this end, tables of stand yield and increment (s z y m k i e w i c z 2001) were used to determine the reduction in the total productivity overbark of stands, as well as – for comparison – the growing stock. the use of total productivity overbark in this case proceeds from an assumption that the process of dieback takes in, not only those trees potentially capable of surviving through to the rotation age (i.e. the main stand), but also those that would be harvested through pre-final felling (i.e. the subordinate stand). it was thereby assumed that data on the quantities of deadwood being removed encompassed both information on the volume of trees that would have been removed by thinning irrespective of the presence or absence of disease, and those that would have formed the mature stand were disease not present. comparison of the volume of deadwood removed throughout the life of the stand with data on total potential productivity overbark in a stand not attacked by disease supplies reliable information as to the theoretical reduction in stock density. ta b l e 2 potential losses of increment in spruce stands of the lg habitat age (y) potential losses of increment overbark (m3/ha/yr) potential current annual increment overbark (m3/ha/yr) reduced increment overbark (m3/ha/yr) reduction in potential increment overbark (%) 25 2.1 9.5 7.4 22 30 2.5 13.3 10.8 19 35 2.9 16.5 13.6 18 40 3.4 18.7 15.3 18 45 3.8 20.4 16.6 19 50 4.2 21.0 16.8 20 55 4.6 20.5 15.9 23 60 5.0 19.5 14.5 26 65 5.5 18.5 13.0 30 70 5.9 17.5 11.6 34 75 6.3 16.6 10.3 38 80 6.7 15.8 9.1 43 85 7.2 15.0 7.8 48 90 7.6 14.2 6.6 53 95 8.0 13.6 5.6 59 100 8.4 12.8 4.4 66 armillaria in the western carpathian mts 225 ta b l e 3 potential losses of increment in spruce stands of the lmg habitat age (y) potential losses of increment overbark (m3/ha/yr) potential current annual increment overbark (m3/ha/yr) reduced increment overbark (m3/ha/yr) % reduction in potential increment overbark 30 8.8 8.8 0 35 11.7 11.7 0 40 14.1 14.1 0 45 15.8 15.8 0 50 16.9 16.9 0 55 17.1 17.1 0 60 16.7 16.7 0 65 15.8 15.8 0 70 15.0 15.0 0 75 0.1 14.3 14.2 0 80 0.2 13.6 13.4 1 85 0.3 12.8 12.5 2 90 0.4 12.1 11.7 3 95 0.5 11.6 11.1 4 100 0.6 10.6 10.0 5 ta b l e 4 potential losses of total productivity overbark in spruce stands of the lg habitat age (y) size of losses overbark (m3/ha) growing stock (m3/ha) total productivity overbark (m3/ha) reduced volume overbark (m3/ha) level of loss in volume overbark (%) reduced total productivity overbark (m3/ha) size of losses to total productivity overbark (%) 25 2 70 71 68 3 69 3 30 14 125 134 111 11 120 10 35 28 189 212 161 15 184 13 40 43 262 305 219 17 262 14 45 61 338 407 277 18 346 15 50 82 410 510 328 20 428 16 55 104 475 611 371 22 507 17 60 128 530 707 402 24 579 18 65 155 574 797 419 27 642 19 70 183 610 883 427 30 700 21 75 214 640 966 426 33 752 22 80 247 666 1047 419 37 800 24 85 282 689 1126 407 41 844 25 90 319 708 1200 389 45 881 27 95 358 723 1269 365 50 911 28 100 400 734 1333 334 54 933 30 230 a. kaliszewski et al. loss borne and the rotation age adopted (there is a shorter period over which the costs incurred prior to the obtainment of income is prolonged, i.e. the stand rotation age achieved). as the figures under discussion and appended tables make clear, the greatest losses were borne in stands of ages 30-40, in the lg habitat, where the rotation age exceeds 100 years – here the abandonment of prophylactic action might mean a loss of around 4600 pln (1 euro ≈ 4 pln) per ha per year. where stands are of rotation ages up to 100 years the value is smaller – up to 900 euro/ha/year. however, there ta b l e 5 potential losses of total, productivity overbark in spruce stands of the lmg habitat age (y) size of losses overbark (m3/ha) growing stock (m3/ha) total productivity overbark (m3/ha) reduced volume overbark (m3/ha) level of loss in volume overbark (%) reduced total productivity overbark (m3/ha) size of losses to total productivity overbark (%) 30 0 71 74 71 0 74 0 35 0 121 130 121 0 130 0 40 0 175 195 175 0 195 0 45 0 234 271 234 0 271 0 50 0 295 354 295 0 354 0 55 0 355 441 355 0 441 0 60 0 407 525 407 0 525 0 65 0 451 605 451 0 605 0 70 0 488 681 488 0 681 0 75 0 519 753 519 0 753 0 80 1 545 821 544 0 820 0 85 2 566 885 564 0 883 0 90 3 583 946 580 1 943 0 95 5 596 1004 591 1 999 1 100 8 606 1059 598 1 1051 1 105 11 614 1111 603 2 1100 1 110 15 620 1160 605 2 1145 1 115 19 624 1205 605 3 1186 2 120 23 627 1247 604 4 1224 2 ta b l e 6 level of potential loss of value in spruce stands of the lg habitat with rotation ages of up to 100 years age subclass age (yr) average age (yr) wu wi wu wi zi zs zi zs loss (m3 of wood) loss (zł) ii a 21-30 25 742.2 348.0 394.2 1.00 0.97 0.03 11.8 1274 ii b 31-40 35 742.2 456.5 285.7 1.00 0.87 0.13 37.1 4000 iii a 41-50 45 742.2 544.1 198.1 1.00 0.85 0.15 29.7 3200 iii b 51-60 55 742.2 611.0 131.2 1.00 0.83 0.17 22.3 2402 iv a 61-70 65 742.2 659.9 82.3 1.00 0.81 0.19 15.6 1684 iv b 71-80 75 742.2 694.7 47.5 1.00 0.78 0.22 10.5 1125 v a 81-90 85 742.2 719.0 23.2 1.00 0.75 0.25 5.8 625 v b 91-100 95 742.2 735.9 6.3 1.00 0.72 0.28 1.8 190 232 a. kaliszewski et al. ł a k o m y p., s i w e c k i r. 2000. gatunki z rodzaju armillaria występujące w nadleśnictwie smolarz. sylwan 4: 115–121. m a n i o n p.d. 1981. tree disease concepts. prentice hall inc., new jersey. p o d g ó r s k i m., b e k e r c., b i c z k o w s k i z., n 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dokumentacja instytutu badawczego leśnictwa. warszawa. z w o l i ń s k i j. 2003. ocena zagrożenia lasów świerkowych w beskidzie śląskim przez zanieczyszczenia powietrza atmosferycznego. prace inst. bad. leśn. 1 (951): 53–68 ż ó ł c i a k a. 1991. zmienność wewnątrz gatunkowa grzybów z rodzaju armillaria – identyfikacja polskich izolatów z rodzaju armillaria. sylwan 11: 27–40. ż ó ł c i a k a. 1999a. identyfikacja gatunków grzybów z rodzaju armillaria (fr.:fr.) staude w polsce. prace inst. bad. leśn. 888: 3–19. żółciak a. 1999b. występowanie grzybów z rodzaju armillaria (fr.:fr.) staude w kompleksach leśnych w polsce. prace inst. bad. leśn. a 890: 29–40. żółciak a. 2003. rozmieszczenie grzybów z rodzaju armillaria w polsce oraz ich rośliny żywicielskie. prace inst. bad. leśn. a 956: 7–22. występowanie i ekonomiczne straty powodowane przez grzyby z rodzaju armillaria we wschodnich karpatach s t r e s z c z e n i e opieńkowa zgnilizna korzeni powodowana przez grzyby rodzaju armillaria stanowi, obok huby korzeni, jeden z najbardziej istotnych problemów ochrony lasu. największe straty ponoszone są przez jednostki alp w drzewostanach iglastych: według danych rdlp powierzchnia drzewostanów w polsce, gdzie stwierdzono szkody spowodowane przez opieńki wyniosła w 1999 roku 144 tys. ha, a już w 2003 roku szacowano ją na ponad 200 tys. ha. choroba występuje w drzewostanach wszystkich klasach wieku zarówno iglastych, jak i liściastych. najbardziej zagrożone są drzewostany w regionach południowych kraju (rdlp katowice i wrocław), w polsce północno-wschodniej (rdlp olsztyn i białystok) oraz w części północno-zachodniej (rdlp szczecin i rdlp toruń). w niektórych rejonach dochodzi do gwałtownego nasilenia wydzielania się posuszu świerkowego, przyjmującego w wielu wypadkach postać rozpadu drzewostanów. dotyczy to w sposób szczególny obszaru beskidu śląskiego armillaria in the western carpathian mts 233 i żywieckiego, gdzie w niżej położonych drzewostanach sytuacja jest katastrofalna. wymuszone tempem zamierania drzew intensywne cięcia sanitarne skutkują postępującym przerzedzeniem drzewostanów, co zwiększa ich podatność na dalsze szkody powodowane przez czynniki abiotyczne (wiatr, śnieg) oraz biotyczne (patogeny, owady kambiofagiczne). w pracy zaprezentowano i przedyskutowano wyniki badań dotyczące analizy ekonomicznej kosztów zabiegów ochronnych i ograniczania strat spowodowanych przez opieńkową zgniliznę korzeni, które mogą dochodzić nawet do 1000 euro z ha. analiza ekonomiczna kosztów związanych z ograniczaniem rozwoju patogenicznych opieniek w drzewostanach uszkodzonych i zagrożonych dostarcza informacji niezbędnych do podejmowania decyzji o konieczności wykonywania zabiegów lub ich zaniechania. pozwolą one na opracowanie strategii polityki ochrony na różnych szczeblach decyzyjnych oraz będą pomocne przy podejmowaniu decyzji odnośnie pilności przebudowy drzewostanów (dostosowania składu gatunkowego do siedliska) lub minimalizacji szkód w nadleśnictwach o dużym zagrożeniu (np. ujsoły, ustroń, wisła). 2014-01-01t11:46:24+0100 polish botanical society a characteristic of mycelium biomass of edible boletus wanda woźniak department of fruit and vegetable technology, institute of food technology the august cieszkowski agricultural university of poznań wojska polskiego 31, pl-60-624 poznań, wozwa@owl.au.poznan.pl w o ź n i a k w.: a characteristic of mycelium biomass of edible boletus. acta mycol. 42 (1):129-140, 2007. the paper presents the results of studies on the production and quality assessment of mycelia of three varieties of king bolete: boletus edulis var. pinicolus vitt. boletus edulis var. piceicolus vasilkov and boletus edulis var. reticulatus (schaeff. ex boud.) bat. in the biomass of mycelium for food the following physicochemical parameters were determined: contents of dry matter, soluble protein – albumins, globulins and prolamins, the rehydratation rate, sensory and microbial quality was assessed. key words: mycelium of edible boletus, quality of mycelium introduction fungi have been a source of food for humans for several thousand years. as a result of deforestation, especially at intensive mushroom harvesting, resources of wild mushrooms in natural habitats have been constantly decreasing. in recent years, applying the advances in biochemical engineering and biotechnology, studies have been initiated on obtaining under controlled conditions of mycelia of large-fruited mushrooms, which could be used as substitutes of fruiting bodies of mushrooms in foodstuffs or as sources of active substances used in medicine ( -� y b e r g e n , s c h e f f e r s 1972; k o h l m ü n z e r 1992; b e a u s é j o u r 1999; b i l a y et al. 2000; l a w 2001). in this way it would be possible to limit the harvesting of mushrooms from natural habitats. �stablishing conditions of mycelium culture in laboratories makes it possible to carry out studies in the field of biology, medicine and food science on all fungus species, irrespective of their seasonal occurrence (� y b e r g e n , s c h e f f e r s 1972; � r z y b e k 1992; k o h l m ü n z e r 1992; � a l l , wa n g 199�; wa s s� r z y b e k 1992; k o h l m ü n z e r 1992; � a l l , wa n g 199�; wa s s � a l l , wa n g 199�; wa s s e r , we i s 1999; ya n g et al. 2 0 0 2 ; wo ź n i a k et al. 2003 a , b ). such investigations were initiated in the second half of the 20th century in the united states by �umbfeld, where at present biomass of morel is sold as flavour substances of this mushroom species (k o p i ń s k i , r o s z a k 19�7; k o p i ń s k i 19�9; m a e k a w a , i n t a b o n 2002). acta mycolo�ica vol. 42 (1): 129-140 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 130 w. woźniak such dried biomass for years has been competing on the american market with dried mushrooms imported from �urope (i l c z u k 19�5; o h t a , f u j i w a r a 2003). production of mycelium biomass of wild mushrooms is an important issue also due to its potential application when new forests are established (k o p i ń s k i 19��; � a l l , wa n g 199�; r y m k i e w i c z 199�; � a l l et al. 2003). in case of some fungi when mycelia are obtained under conditions of controlled production of submerged cultures, they may be cultivated, as it is the case e.g. with the cultivation of white truffle in new zealand (ve l l i n g a 2003). mat�rials and m�t�ods the adopted objective of the study was to obtain mycelium for food production purposes from selected species of wild mushrooms characterized by large fructifications, attractive for consumers. in this study the following investigations were conducted: 1. obtaining matrix mycelium from fruiting bodies of such mushrooms as: boletus edulis var. pinicola vitt., king bolete ‘pine’ boletus edulis var. piceicola vasilkov, king bolete ‘spruce’ boletus edulis var. reticulatus (schaeff. ex boud.) bat, king bolete ‘reticulated’ 2. production of mycelium for food production purposes under laboratory conditions using submerged culture. 3. physicochemical, sensory and microbial quality assessment of mycelium for food production purposes. the characteristic of experimental material raw material for the production of matrix mycelium were wild mushroom fructifications growing in their natural habitats. king bolete boletus edulis bull.: fr. of three varieties were collected in the milickie forests near łazy wielkie and in the tucholskie forests near brusy. fruiting bodies of mushrooms, from which tissue matrix mycelium was collected, in terms of their quality and morphological characters could be classified as extra quality class according to the polish standard pn-76r7�505 for fresh mushrooms. fructifications were young, healthy and all the mushrooms exhibited characters characteristic of a given species and variety (wo ź n i a k , wą s o w i c z 19�1). pileus surfaces were not damp, they were properly coloured. mushrooms had whitish hymenophores with immature spores. all the mushrooms had a typical aroma. the flesh at intersection was whitish, firm and fleshy (� r ű n e r t , � r ű n e r t 19�4; l a e s s ø e , c o n t e 1997). matrix mycelium obtained from these mushrooms was used a raw material in the production of mycelium for food production purposes. the mycelium was produced and its quality was assessed in the years 199�–2004. methods mycelium production. �rowth of mycelium on commercial agar solidified media. matrix mycelium was obtained using tissue culture from wild growing mushrooms. reproduction of mycelium for food production purposes was produced on mycelium biomass 131 potato and wheat media. in case of experimental mycelia mycelium growth increments were established on petri dishes. during mycelium growth the appearance of the mycelium was assessed descriptively in sensory analysis. mycelium proliferation using submerged liquid cultures. proliferation was conducted in sterile liquid media, each time in 16 replications. media were spawned with reproduction mycelium. after spawning flasks were sealed with sterile cotton wool stoppers and placed in a 35� s shaker (fig. 1). prolification parameters were established experimentally (wo ź n i a k 2002; wo ź n i a k , k o r z e n i e w s k a 2003). constant p� and extract were maintained all that time in the medium. �rowth dynamics was determined as a percentage increment of the liquid volume in reaction flasks taken up by the mycelium. the structure of mycelial filaments, the shape and colour of the mycelium were assessed visually and described every day, starting from the 2nd day after spawning. tissue mycelia obtained using the traditional method were stored on petri dishes, sterile slants and sterile wheat grain (fig. 2). mycelia obtained in submerged cultures after the completion of proliferation were drained on the �4 filter, thoroughly washed with water and carefully filtered and surface dried. mycelia for food were dried in a laboratory desiccator with enforced air circulation at 35°c and force dried at 55°c. the obtained dried mushrooms were packed in polyehtylene bags and next into glass jars and stored at 20–22°c, relative humidity of 35% in the dark (fig. 3). methods of quality appraisal of mycelium for food. in the biomass of mycelium for food were determined: physicochemical parameters, sensory and microbial quality was assessed. – determination of dry matter content using the scaling method (c h a r ł a m p o w i c z 1966). – determination of contents of soluble protein compounds (in �2o, in 0.1 m nacl, in 15% nao�) using the colorimetry method (m a j b a u m k a t z e n e l l e n b o g e n , m o c h n a c k i 196�; wo ź n i a k 19�3). – �ot determination of the rehydration coefficient in dried mycelium using the loesecke method (c h a r ł a m p o w i c z 1966). – determination of microbial purity (total microbial counts, levels of coliform bacteria, e. coli counts, contents of yeasts, moulds, the presence of coagulase-positive staphylococci) using the platelet and test methods [pn-93 a-�6034/02; pn-90 a-75052/04; pn-90 a-75052/05; pn-90 a-75052/0�; pn-�n-iso 6���-2;] – determination of aromatic compounds using the spm� method, gas chromadetermination of aromatic compounds using the spm� method, gas chroma-determination of aromatic compounds using the spm� method, gas chromatography, with the identification of isolated compounds using mass spectrophotometry (t h o m a s 1973; wą s o w i c z , k a m i ń s k i 1974). – sensory analyses of mycelia, using a 5-point scale according to tilgner (b a r y ł k o -p i k i e l n a 1975). – obtained results of conducted physico-chemical and sensory analyses, as well as mycelium growth dynamics were interpreted statistically. the stat 6 by statsoft software was used for the calculations. to illustrate mycelium growth dynamics the equation of the constant mycelium growth was applied : kp� = r = ∂ [ø] / ∂ t;, in which kp� is the mycelium growth constant, r – function of trend, ∂ [ø] – change in the thallus area in mm, ∂ t. – change in the time of mycelium growth increment in days (m c � w a n 1991; b r a n d t 2002). 132 w. woźniak r�sults mycelium for food was obtained from the second reproduction of matrix mycelium, on potato and wheat media. these media in earlier studies gave good production yields and were asessed sensorily as the best (wo ź n i a k et al. 2000; wo ź n i a k 2002; wo ź n i a k et al. 2002; wo ź n i a k et al. 2003a, b; wo ź n i a k , k o r z e n i e w s k a 2003; wo ź n i a k et al. 2001; wo ź n i a k et al. 2004; k o r z e n i e w s k a , wo ź n i a k 2005). the production of mycelium for food was completed at the moment of the phase of growth inhibition in reaction chambers. physico-chemical analyses were conducted immediately after production, after washing with distilled water and surface drying of mycelium. the analysis of aroma was conducted for fruiting bodies of fresh mushrooms, from which matrix mycelium was obtained. for the mycelium aroma was assessed between day 5 and day 10 of proliferation depending on the variety of the analyzed mushrooms. sensory analyses were performed for fresh mycelium, dried mycelium and mycelium after rehydration. the appaearance (colour and consistency), taste and aroma were assessed. for dried and ground mycelium for food the analysis of microbial purity was conducted in the range of analyses required for dried mushrooms. the production of mycelium for food was completed at the moment of the phase of growth inhibition in reaction chambers. mycelia of all varieties grew faster on potato medium. proliferation of matrix mycelium lasted approx. 64 days for the “spruce” variety, 74 days for the “pine” variety and 76 days for the “reticulated” variety. the production of biomass in case of mycelium for food lasted much shorter and thus it was 12 days for “pine” variety on potato medium and 14 days on wheat medium, for the “reticulated” variety it was � days on potato medium and 10 days on wheat medium. mycelium, irrespective of the medium type, grew unformly in the form of hyphae, which constituted branching threads (tabs 1-2, figs 4-9). mycelia of king bolete in comparison to fruiting bodies had comparable contents of dry matter. depending on the medium used for proliferation, in mycelia obtained from potato and wheat media the solids content for king bolete ranged ta b l e 1 constant kp� for growth changes in matrix mycelium medium constant kp� for king boletus ‘spruce’ ‘pine’ ‘reticulated’ wheat 1.�6 1.16 1.�1 potato 2.14 1.54 1.93 ta b l e 2 constant kp� for growth cymelium in submerged liquid method medium variety of king boletus ‘spruce’ ‘pine’ ‘reticulated’ wheat 7.40 14.63 �.54 potato 9.29 1�.06 9.71 mycelium biomass 135 fractions for mycelia of the same variety differed depending on the medium used for proliferation. in mycelia grown on potato medium the content of soluble protein was slightly higher than in mycelia obtained from wheat medium. the differences amounted to 2.4% for ‘spruce’ king bolete, 2.�% for ‘pine’ king bolete and 4.3% for ‘reticulated’ king bolete. contents of albumins ranged from 52% to 63% soluble protein contents, while those of globulins from 22.7% to 36% and prolamins from �.3% to 15.6%, respectively (tabs 3-5). the hot determination of the rehydration ratio was conducted for 20 minut. the coefficient was high and amounted to approx. 6 for ‘spruce’ and ‘pine’ king bota b l e 3 contents of selected chemical contents in mycelium of ‘spruce’ king boletus analysis contents mean (n=16) minimum maximum coefficient of variation multiplying on potato medium dry matter (%) �.53 �.42 �.62 1.17 (g/100g s.s.) total solube protein albumins globulins prolamins 33.49 1�.94 11.39 3.16 33.65 1�.74 11.25 2.9� 33.31 19.0� 11.59 3.32 1.15 multiplying on wheat medium dry matter (%) 9.14 9.04 9.22 0.76 (g/100g s.s.) total solube protein albumins globulins prolamins 32.71 1�.5� 11.41 2.72 32.57 1�.45 11.26 2.60 32.�5 1�.71 11.52 2.�6 1.67 n = number of samples collected for analyses ta b l e 4 contents of selected chemical contents in mycelium of ‘pine’ king boletus analysis contents mean (n=16) minimum maximum coefficient of variation multiplying on potato medium dry matter (%) �.25 �.10 �.32 1.21 (g/100g s.s.) total solube protein albumins globulins prolamins 32.�4 17.�9 11.47 3.4� 32.75 17.7� 11.42 3.41 32.�9 17.92 11.56 3.55 0.74 multiplying on wheat medium dry matter (%) �.07 �.01 �.13 0.3� (g/100g s.s.): total solube protein albumins globulins prolamins 31.53 16.30 10.�� 4.35 31.13 16.09 10.79 4.25 31.79 16.31 11.01 4.47 1.21 n = number of samples collected for analyses 13� w. woźniak amounted in the fruiting body of ‘spruce’ and ‘pine’ king bolete approx. 99% and in the ‘reticulated’ variety 72% total volatile compounds. in mycelia of these species it was 40% for ‘pine” king bolete, 29% for ‘spruce’ variety and �% for ‘reticulated” variety, respectively (tab. �). conclusions problems investigated in this study concern the production of high quality mycelium of king bolete of three varieties: ‘pine’, ‘spruce’ and ‘reticulated’. in the conducted investigations it was found that: 1. composition of the medium applied for proliferation has a significant effect on growth dynamics of mycelium and on its quality. 2. physico-chemical indexes (protein contents and dry matter, aromatic volatile substances), sensory attributes (appearance, taste and aroma) and microbial indexes indicate high quality of the produced mycelia. 3. mycelia produced from the analyzed mushroom varieties after drying may constitute a good supplement of dried mushroom in foodstuffs. ta b l e 7 description of biomass for food of king bolete varietas medium quality attributes colour taste aroma ‘spruce’ potato light-cream mushroom mushroom intensive wheat light-cream-white mushroom mushroom, average intensive ‘pine’ potato light-cream mushroom mushroom intensive wheat light -cream-white small mushroom small mushroom ‘reticulated’ potato cream mushroom mushroom wheat cream-beige small mushroom small mushroom ta b l e � aroma components in mycelium and fruting bodies contents of aroma components in ppm king bolete volatile componends �carbon compounds 1-octen-3-ol ‘spruce’ fruting 1 64�4 6329 6293 mycelium 2 2�30 24�� 725 ‘pine’ fruting 1 11496 10974 10�16 mycelium 2 2235 1600 642 ‘reticulated’ fruting 1 205�4 15375 11052 mycelium 2 3662 2�20 222 �xplanations: 1 aroma components of fresh fruting bodies analyzed for mushrooms from which matrix mycelium was obtained. 2 aroma components of mycelium biomass obtained in submerged liquid cultures on potato medium on day 5 of proliferation. mycelium biomass 139 r�f�r�nc�s b a r y ł k o p i k i e l n a n. 1975. zarys analizy sensorycznej żywności. wnt, warszawa. b e a u s é j o u r t. m. 1999b. �etting started with mushroom cultivation. www.mycoweb.com/articels/ cultivation.html. b i l a y v. t. , s o l o m k o � . 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shiitake odmiany 37*37 z hodowli wgłębnej. roczniki ar w poznaniu, cccxxiii. ogrodnictwo 31 (1): 561–565. w o ź n i a k w. , m u r a s u . , k o r z e n i e w s k a a . , � a p i ń s k i m . 2001. �rowth of agaricus bisporus (lange) sing. mycelium as influenced by production method. vegetable crops res. bull., poland-skierniewice 54 (2): �3–�6. w o ź n i a k w. 2002. raport z badań nad produkcją grzybni wielkoowocnikowych grzybów w latach 19972002. materiały niepublikowane, do wglądu w itżpr ar w poznaniu. w o ź n i a k w. , m u r a s u . , k o r z e n i e w s k a a . , � a p i ń s k i m . 2002. opracowanie warunków produkcji grzybni pieczarki dwuzarodnikowej odmiany polmycel 3201 metodą wgłębną. materiały xxxiii sesji naukowej ktichż pan „nauka o żywność. osiągnięcia i perspektywy”. 10-11 wrzesień 2002, lublin: 270. w o ź n i a k w. , � a p i ń s k i m . , m u r a s u . , k o r z e n i e w s k a a . 2003a. ocena jakości grzybni shiitake odmiany s� 21 wyprodukowanej metodą wgłębną. materiały ii krajowego kongresu biotechnologii, 23-27 czerwiec 2003, łódź: 21. w o ź n i a k w. , � a p i ń s k i m . , k o r z e n i e w s k a a . , m u r a s u . 2003b. wzrost i charakterystyka biomasy grzybni soplówki. folia �orti., 1: 142–145. w o ź n i a k w. , k o r z e n i e w s k a a . 2003. produkcja biomasy i ocena grzybni borowika szlachetnego. folia �orti. 1: 349–351. w o ź n i a k w. , m u r a s u . , k o r z e n i e w s k a a . 2004. wpływ przechowywania grzybni borowika szlachetnego boletus edulis (fries) na jej żywotność. folia univ. agric. stetin., agricultura 95: 425–429. ya n g j . � . , l i n � . c h . , m a u j . l . 2002. antioxidant properties of several commercial mushrooms. food chem. 77: 229–235. pn-76 r-7�505 – �rzyby świeże. pn-93 a-�6034/02 – badania mikrobiologiczne – ogólne zasady badań. pn-90 a-75052/04 – metody badań mikrobiologicznych. sposób pobierania i przygotowanie próbek do badań mikrobiologicznych. pn-90 a-75052/05 – metody badań mikrobiologicznych. oznaczanie obecności i liczby drobnoustrojów tlenowych mezofilnych i psychrofilnych. pn-90 a-75052/0� – metody badań mikrobiologicznych. oznaczanie liczby drożdży i pleśni. pn-�n-iso 6���-2 – mikrobiologia żywności i pasz – �oryzontalna metoda oznaczania liczby gronkowców koagulazo-dodatnich (staphylococcus aureus i innych gatunków) charakterystyka biomasy grzybni borowika szlachetnego s t r e s z c z e n i e wykorzystując osiągnięcia inżynierii biochemicznej i biotechnologii, rozpoczęto w ostatnich latach badania nad otrzymaniem grzybni grzybów wielkoowocnikowych w warunkach kontrolowanych, którą można byłoby wykorzystać jako zamienniki owocników grzybów w produktach spożywczych. w ten sposób ograniczyć można byłoby zbiór grzybów ze środowisk naturalnych dla przetwórstwa. publikacja przedstawia wyniki badań nad oceną grzybni dla celów spożywczych z wybranych odmian atrakcyjnego gatunku boletus edulis. w tym opracowaniu przedstawiono badania dotyczące otrzymania grzybni matecznej i spożywczej owocników następujących grzybów: borowik szlachetny ‘sosnowy’ boletus edulis var. pinicolus vitt., borowik szlachetny ‘świerkowy’ boletus edulis var. piceicolus vasilkov, borowik szlachetny ‘usiatkowany’ boletus edulis var. reticulatus (schaff ex. boud) bat. �rzybnię wyprodukowano w warunkach laboratoryjnych metodą wgłębną na wytrząsarce shaker typ 35�s, na pożywce pszennej i ziemniaczanej. ocena jakości grzybni obejmowała współczynniki: fizykochemiczne, sensoryczne oraz mikrobiologiczne. w przeprowadzonych badaniach stwierdzono, że pozyskiwane grzybnie były wysokiej jakości. wyróżniki fizykochemiczne: zawartość białka i sucha substancja oraz substancje lotne aromatu; sensoryczne: wygląd, smak i zapach oraz mikrobiologiczne, wskazują na wysoką jakość otrzymywanych grzybni. skład fizykochemiczny grzybni zależał w sposób istotny od odmiany borowika i zastosowanej pożywki. fig. 1. biomass of matrix mycelium transmitted on grain. fig. 2. mycelium of boletus obtained in submerged liquid culture. fig. 3. dried biomass of mycelium for food. 2014-01-01t11:45:46+0100 polish botanical society arbuscular mycorrhizal fungi (glomeromycota) associated with roots of plants of the lubuskie province sławomir kowalczyk and janusz błaszkowski department of plant protection, west pomeranian university of technology słowackiego 17, 71-434 pl szczecin, janusz.blaszkowski@zut.edu.pl kowalczyk s., błaszkowski j.: arbuscular mycorrhizal fungi (glomeromycota) associated with roots of plants of the lubuskie province. acta mycol. 46 (1): 3–18, 2011. the results of studies of the occurrence of arbuscular mycorrhizal fungi (amf) and arbuscular mycorrhizae of the phylum glomeromycota associated with roots of 31 cultivated, uncultivated and protected plant species growing at 103 sites of the lubuskie province nw poland are presented and discussed. the amf most frequently found were members of the genus glomus. other relatively frequently revealed fungi were scutellospora spp. spore populations of amf generally were more abundant and diverse in cultivated soils. most protected plant species harboured amf. key words: distribution, occurrence, cultivated, wild and protected plants, poland introduction arbuscular mycorrhizal fungi (amf) of the phylum glomeromycota (schüßler, schwarzott and walker 2001) belong to the most widely distributed soil microorganisms in the world and are associated with ca. 70-90% of vascular land plants (smith, read 2008). at present, the phylum glomeromycota consists of four orders, 10 families and 14 genera (oehl, sieverding 2004; palenzuela et al. 2008; schüßler et al. 2001; sieverding, oehl 2006; walker et al. 2007). the most numerous group within the glomeromycota is the genus glomus, comprising ca. 53% of all species of the phylum known to date, i.e., ca. 220 (błaszkowski 2003). amf are well known to increase productivity and vigour of plants, as well as their resistance to different abioand biotic stresses (schönbeck 1978; koske et al. 2004). additionally, amf stabilize soils and improve their structure throughout binding sand grains and aggregate formation (koske, polson 1984). the effectiveness and stability of such influences are generally higher when amf communities are more diverse (bever et al. 1996; klironomos et al. 2000). acta mycologica vol. 46 (1): 3–18 2011 4 s. kowalczyk and j. błaszkowski asexual spores of amf are persistent propagules that remain infectious in the absence of host plants and under unfafourable conditions, e.g., because of influences of different agricultural practices. on the other hand, different plant species and farming practices may variously affect the spore production of different species of amf (bever et al. 1996; jansa et al. 2002; oehl et al. 2009). recognition of the reasons that mainly affect amf may be used in constructing of crop rotations and farming practices to retain high vitality of this important group of soil microorganisms. most studies of amf diversity rely on morphological identification of spores extracted either from field-collected soil samples or/and trap cultures in which field soils with appropriate host plants are grown in a greenhouse (oehl et al. 2009). however, each of these techniques has constrains. examination of field soils frequently reveals amf that do not sporulate in trap cultures (błaszkowski, pers. observ.). on the other hand, cultivation of field soils in trap cultures may initiate sporulation of fungi forming spores seasonally, rarely or not at all in field conditions (błaszkowski, kovács and balázs 2009; błaszkowski, tadych and madej 2000; błaszkowski et al. 2009; stutz, morton 1996). an ideal method of recognizing of amf associated with plant roots seems to be that using molecular tools. however, its high costs and labor, as well as the lack of specific molecular markers for most existing amf also restrict their wide application (öpik et al. 2009). one of the main aims of numerous plant protection projects are the preservation and protection of rare and endangered plant species (zubek, turnau and błaszkowski 2005). most of these plants probably co-occur with amf, although the amount of literature data on this subject is very low. the recognition of amf most frequently co-existing with such plant species may be used to protect them by introduction of these fungi into sites where the plants grow. in soils of the lubuskie province, only one species of amf has been found to date, i.e., acaulospora thomii błaszk. (błaszkowski 1988). therefore, the aim of this study was to better know amf associated with roots of cultivated, uncultivated (not protected) and protected plants growing in soils of this region. materials and methods the study material and area. the study material were mixtures of rhizosphere soils and roots collected under 31 cultivated, wild (not protected), and protected plant species growing at 103 sites located in the lubuskie province (fig. 1). collection of rhizosphere soils and roots, establishment and growth of trap cultures, extraction of spores, staining of mycorrhizae, and identification of amf. rhizosphere soils and root fragments were collected from may to august of the years 2003-2006. they were taken from a depth of 5-30 cm using a small garden shovel and then placed in plastic bags. in the laboratory, they were air dried and then stored in a refrigerator at ca. 4oc for 1-4 months. trap cultures were established to obtain a large number of living spores and to initiate sporulation of species that were present but were not detected in the field collections (stutz, morton 1996). the method used to establish trap cultures, their 6 s. kowalczyk and j. błaszkowski 132; beta vulgaris l.: 1; 2; 3; 28; 29; 30; 68; 69; 71; rogoziniec; kaława; nowa wioska; brudzewo; babimost; zbąszynek; krzepielów; żabice; grochowo; 22.07.2003; 2.08.2003; -“-; 12.08.2004; -“-; -“-; 18.06.2005; 19.06.2005; -“-; 1; 2; 3; 54; 55; 56; 133; 134; 135; hordeum vulgare l.: 1; 1; 7; 8; 29; 29; 34; 35; 75; 76; 77; rogoziniec; -“-; wysoka; bukowiec; babimost; -“-; skąpe; pieski; lubomyśl; różanówka; leśniki; 22.07.2003; -“-; 2.08.2003; 3.08.2003; 12.08.2004; -“-; -“-; 12.08.2004; 18.06.2005; -“-; -“-; 1; 7; 8; 9; 55; 61; 60; 62; 139; 140; 141; secale cereale l.: 9; 10; 11; 36; 37; 38; 78; 79; 80; samsonki; stary dwór; lutol suchy (st. kolejowa); międzyrzecz; połupin; sycowice; gorzupia; dzietrzychowice; dęby; 22.07.2003; 2.08.2003; 3.08.2003; 12.08.2004; -“-; -“-; 18.06.2005; 18.06.2005; -“-; 10; 11; 12; 63; 64; 65; 142; 143; 144; solanum tuberosum l.: 4; 40; 47; 48; 49; 50; 90; 91; 92; dąbrówka wlkp.: łazy; pomorsko; smardzewo; podmokle małe; kosieczyn; surowa; krążkowo; wrociszów; 12.08.2004; -“-; -“-; -“-; -“-; -“-; 18.06.2005; -“-; -“-; 80; 75; 76; 77; 78; 79; 154; 155; 156; triticum aestivum l.: 8; 10; 14; 15; 32; 32; 39; 81; 82; 83; bukowiec; stary dwór; wysoka; brójce; brody; -“-; czerwieńsk; jutrzenka; kierzno; styrułów; 12.08.2004; 2.08.2003; -“-; 3.08.2003; 12.08.2004; -“-; -“-; 18.06.2005; 18.06.2005; -“-; 66; 17; 16; 18; 58; 68; 67; 145; 146; 147; xtriticose-cale wittmack: 12; 13; 41; 42; 43; 84; 85; 86; lutol suchy; szumiąca; łagów (k/gronowa); łęgowo; smardzewo; lubogoszcz; podbrzeże; dębianka; 22.07.2003; 2.08.2003; 12.08.2004; -“-; -“-; 18.06.2005; -“-; -“-; 13; 14; 69; 70; 71; 148; 149; 150; zea mays l.: 16; 17; 18; 44; 45; 46; 87; 88; 89 staropole; boroszyn; myszęcin; przytoczna; rubinów; chociule; stare strącze; bieniów; drogomin; 2.08.2003; -“-; -“-; 11.08.2004; 12.08.2004; -“-; 18.06.2005; -“-; 19.06.2005; 19; 20; 21; 72; 73; 74; 151; 152; 153. uncultivated (not protected) plants. achillea millefolium l.: 2; 19; 20; 51; 52; 53; kaława; chociszewo; jasieniec; lubrza; romanówek; bucze; 2.08.2003; 22.07.2003; 2.08.2003; 13.08.2004; -“-; -“-; 23; 22; 24; 81; 82; 83; cirsium arvense l.: 21; 22; 23; 51; 54; 55; wilenko; wityń; żydowo; lubrza; jemiołów; zarzyń; 3.08.2003; -“-; 3.08.2003; 13.08.2004; -“-; -“-; 25; 26; 27; 84; 85; 86; melandrium album l.: 51; 52; 56; lubrza; romanówek; łagów; 13.08.2004; -“-; -“-; 87; 88; 89; sedum maximum (l.) hoffm.: 26; 26; 51; 59; 95; 96; 97; trzciel; -“-; lubrza; żelechów; rzepin; torzym; górzyce; 3.08.2003; 15.08.2004; -“-; -“-; 19.06.2005; -“-; -“-; 32; 95; 81; 94; 161; 160; 162; carex sylvatica huds: 57; 58; 93; 94; 95; rybojady; borowy młyn; drezdenko; ośno lubuskie; rzepin; 15.08.2004; -“-; 18.06.2005; 19.06.2005; -“-; 90-91; 92; 157; 158; 159; equisetum arvense l.: 5; 25; 26; 55; 56; 57; sierczynek; bieleń; trzciel; łagów; rybojady; 3.08.2003; -“-; 3.08.2003; 13.08.2004; -“-; -“-; 33; 31; 32; 96; 97; 98; trifolium arvense l.: 2; 53; 54; 64; 70; 98; kaława; bucze; jemiołów; jabłonów; drzecin; jagielniki; 12.08.2004; 13.08.2004; -“-; 18.06.2005; 19.06.2005; 18.06.2005; 99; 100; 101; 164; 163; 165; hypericum perforatum l.: 51; 52; 54; 96; 99; 100; lubrza; romanówek; jemiołów; torzym; nowa sól; dobiegniew; 13.08.2004; -“-; -“-; 19.06.2005; 18.06.2005; 19.06.2005; 102; 103; 104; 168; 166; 167; juncus effusus l.: 26; 56; trzciel; łagów; 15.08.2004; 21.06.2005; 105-107; 169-171; plantago arenaria waldst. et kit.: 26; 26; trzciel; -“-; 15.08.2004; 22.06.2005; 108-110; 172-174; plantago lanceolata l.: 16; 20; 23; 54; 57; 60; staropole; jasieniec; żydowo; jemiołów; rybojady; siedlisko; 2.08.2003; 3.08.2003; -“-; 13.08.2004; -“-; -“-; 34; 35; 36; 113; 111; 112; corynephorus canescens (l.) p. beauv.: 26; 94; 101; 102; trzciel; ośno lubuskie; stary jaromierz; skólsko; 15.08.2004; 19.06.2005; 18.06.2005; -“-; 114-116; 176; 175; 177; polygonum persicaria l.: 5; 13; 23; 58; 61; 62; sierczynek; szumiąca; żydowo; borowy młyn; siercz; świdwowiec; 3.08.2003; 2.08.2003; 3.08.2003; 15.08.2004; -“-; -“-; 39; 37; 38; 117; 118; 119; rumex acetosella l.: 4; 6; 17; 37; 52; 63; dąbrówka wlkp.; łagowiec; boroszyn; połupin; romanówek; gronów; 22.07.2003; 2.08.2003; -“-; 12.08.2004; 13.08.2004; 12.08.2004; 40; 41; 42; 120; 122; 121; potentilla anserina l.: 26; 103; trzciel; słońsk; 15.08.2004; 19.06.2005; 123-125; 178-180. protected plants. vinca minor l.: 56; 56; łagów; -“-; 21.06.2005; 24.06.2006; 217-218; 226-228; hedera helix l.: 56; 56; łagów; -“-; 13.08.2008; 21.06.2005; 211-213; 219-223; helichrysum arenarium (l.) moench: 26; 26; trzciel; -“-; 3.08.2003; 15.08.2004; 181-185; 196-198; jovibarba sobolifera (sims.) opiz: 26; 26; trzciel; -“-; 3.08.2003; 15.08.2004; 186-190; 199-201; convallaria majalis l.: 56; 56; 56; łagów; -“-; -“-; 17.04.2004; 9.04.2005; 8.04.2006; 205-207; 214-216; 224-225; lycopodium clavatum l.: 26; 26; trzciel; -“-; 3.08.2003; 15.08.2004; 191-195; 202-204. *after the plant species name, the number of location(s), the name of location(s), the date(s) of collection, and the number of samples are listed, respectively. arbuscular mycorrhizal fungi 7 growing conditions, and the methods of spore extraction and staining of mycorrhizae were as those described previously (błaszkowski, renker and buscot 2006). the host plants were plantago lanceolata and zea mays. morphological properties of spores and their subcellular structure were determined based on examination of at least 50 spores mounted in water, lactic acid, polyvinyl alcohol/lactic acid/glycerol (pvlg; omar, bollan and heather 1979), and a mixture of pvlg and melzer’s reagent (1:1, v/v). spores at all developmental stages were crushed to varying degrees by applying pressure to the cover slip and then stored at 65o c for 24 h to clear their contents from oil droplets. they were then examined under an olympus bx 50 compound microscope equipped with nomarski differential interference contrast optics. microphotographs were recorded on a sony 3cdd color video camera coupled to the microscope. amf were identified according to original descriptions, specimens collected by j. błaszkowski, and descriptions and illustrations presented in błaszkowski (2003) and morton (2002). chemical and statistical analyses. for chemical analyses, 81 soil samples were selected, in which ph (in 1n kcl), the contents of n, p, k, organic c (in g.kg-1 of dry matter), and available forms of p, k, and mg (in mg.kg-1 of dry matter) were determined. each cultivated and uncultivated (not protected) plant species was represented by three randomly selected soil samples, and each protected plant species by one. differences in the structure of amf communities were investigated by determining the frequency of occurrence of species, spore abundance and species richness, and by calculating dominance coefficients (górny, gruma 1981). spore abundance and coefficients of dominance were determined based on spores isolated only from field-collected samples. frequency of occurrence and species richness were calculated based on spores isolated from both field-collected samples and trap cultures. frequency of occurrence was calculated by determining the percentage of fieldcollected samples and trap cultures from which spores of a particular species were recovered. spore abundance and species richness were defined by determining the number of spores and species, respectively, occurring in 50 or 100 g dry soil. dominance coefficient expresses the proportion of the number of spores of a particular species in all spores of amf recovered. coefficients of correlation were used to determine relationships between the spore abundance and soil chemical properties. results and discussion arbuscular mycorrhizal fungi and arbuscular mycorrhizae associated with cultivated and uncultivated (not protected) plants arbuscular mycorrhizal fungi. the occurrence of amf in cultivated and uncultivated soils was determined based on spores isolated from field-collected rhizosphere soil-root samples and trap cultures established from a part of each field sample. the soil-root samples came from under 25 plant species belonging to 16 families (fig. 1). 8 s. kowalczyk and j. błaszkowski of them 10 were cultivated plant species, and the others uncultivated once, including medicine plants. each cultivated plant species was represented by 9 soil-root samples, and uncultivated one by 6. a total of 40287 spores of amf were isolated, including 11517 spores from field samples (of which 39.6% come from under cultivated plants) and 28770 spores from trap cultures. the spores represented 9 of the 14 exiting genera of the glomeromycota (tab. 1). most species (18) were from the genus glomus. the genus acaulospora was represented by 5 species, and the genera entrophospora, gigaspora, pacispora, and scutellospora by 5 species each. one species each came from the genera ambispora and paraglomus. additionally, not numerous spores of unrecognized species of the genera gigaspora, glomus, and scutellospora were found. occurrence of amf. spores of amf occurred in 92.8% of field-collected soil samples. they represented 13 species of glomus, 5 of acaulospora, 3 of scutellospora, 2 each of entrophospora, gigaspora and pacispora, and 1 of ambispora (tab. 1). in the root zone of cultivated plants, 15 species in three genera occurred. roots of wild plants harboured 28 species in 7 genera and not numerous spores of unrecognized species of glomus and scutellospora. of the am fungal species identified in field soils, 21 sporulated in trap cultures (tab. 1). spores of acaulospora capsicula, ac. lacunosa, ac. koskei, ac. mellea, entrophospora baltica, gigaspora gigantea, and glomus fuegianum were revealed only in field soils. in trap cultures, 27 species and not numerous spores of unrecognized morphotypes were revealed. the amf most frequently occurring in the lubuskie province soils were members of the genus glomus (tab. 1); they occurred in 87.2% of soils, of which 47.0% represented cultivated plants. glomus spp. were associated with all plant species of the families asteraceae, caryophyllaceae, cyperaceae, equisetaceae, hyperiaceae, and polygonaceae. other relatively frequently revealed amf were scutellospora spp. (more frequently co-occurred with most families of uncultivated plants) and pacispora spp. (were more frequently harboured by families of cultivated plants). members of the genera acaulospora, ambispora, gigaspora, and paraglomus were recorded only in soils from under uncultivated plants (tab. 1). the disclosure of 10 species and one undescribed morphotype in trap cultures that were not found in field soils confirms conclusions of, e. g., błaszkowski, tadych and madej (2002), stütz & morton (1996) and jansa et al. (2002) that a large part of amf may not sporulate in the field at all or their sporulation is seasonal. frequency of occurrence of species. the species most frequently occurring in cultivated soils of the lubuskie province were gl. mosseae (present in 80.0% of soils) and gl. claroideum (62.2%; tab. 1). fungi relatively frequently recorded (present in 15-30% of soils) also were gl. caledonium and p. franciscana. in uncultivated soils, the most frequently found fungi included gl. constrictum (51.1%) and gl. claroideum (44.0%), followed by ac. lacunosa, gl. macrocarpum, and s. dipurpurescens. dominance. the eudominants (of a coefficient of dominance of d>20%) of cultivated sites were gl. claroideum and gl. deserticola (tab. 2). the group of dominants (d=10-20%) formed only gl. mosseae. the subdominants (d=5-10%) were gl. caledonium, p. scintillans, and s. dipurpurescens. in uncultivated soils, the eudominants were gl. constrictum and gl. claroideum. glomus badium, gl. deserticola, gl. lamel arbuscular mycorrhizal fungi 9 losum and s. dipurpurescens were dominants, and e. infrequens, gl. caledonium, and gl. mosseae subdominats. the data presented above confirm błaszkowski’s (1993) and gerdemann’s (1968) conclusions that amf are common soil fungi and coexist with most vascular cultivated and uncultivated plant species of the world. the abundant and diverse spore populations of glomus spp. revealed in the study discussed here indicate a good adaptation of these fungi to a wide range of soil conditions (anderson, liberta and dickman 1984; grey 1991; jansa et al. 2002; porter, robson and abbott 1987). species of gigaspora and scutellospora prefer warmer table 1 frequency of occurrence of amf isolated from under cultivated (a) and uncultivated (not protected) plants (b) of the lubuskie province frequency of occurrence (%) fungus trap cultures with field soils p. lanceolata z. mays a b a b a b acaulospora capsicula 5.56 acaulospora lacunosa 16.67 acaulospora koskei 1.11 acaulospora mellea 4.44 acaulospora paulinae 1.11 1.11 ambispora gerdemannii 13.33 2.22 archaeospora trappei 2.22 2.22 4.44 entrophospora baltica 1.11 entrophospora infrequens 1.11 5.56 3.33 4.44 2.22 gigaspora gigantea 1.11 gigaspora margarita 2.22 4.44 2.22 gigaspora sp. 1.11 glomus aggregatum 5.56 10.00 3.33 11.11 2.22 2.22 glomus badium 1.11 8.89 3.33 1.11 glomus caledonium 17.78 5.56 37.8 10.00 30.00 17.78 glomus claroideum 27.78 20.00 56.7 44.44 62.22 40.00 glomus clarum 5.56 glomus constrictum 28.89 51.11 11.1 26.7 1.11 8.89 glomus deserticola 24.44 25.56 15.6 10.00 1.11 glomus fasciculatum 4.44 12.22 2.22 4.44 2.22 glomus fuegianum 1.11 glomus geosporum 3.33 5.56 2.22 glomus lamellosum 1.11 7.78 1.11 6.67 glomus macrocarpum 8.89 16.67 1.11 glomus microcarpum 1.11 6.67 1.11 1.11 glomus mosseae 80.00 26.67 61.1 31.11 60.00 25.56 glomus pansihalos 1.11 glomus pustulatum 1.11 1.11 glomus rubiforme 1.11 1.11 glomus verruculosum 1.11 2.22 glomus 178 3.33 4.44 2.22 7.78 glomus sp. 1.11 1.11 2.22 1.11 2.22 pacispora franciscana 16.67 8.89 11.1 3.33 10.00 3..33 pacispora scintillans 12.22 7.78 8.89 3.33 6.67 2.22 paraglomus laccatum 1.11 4.44 scutellospora armeniaca 1.11 1.11 1.11 scutellospora dipurpurescens 8.89 23.33 8.89 17.78 5.56 14.44 scutellospora pellucida 6.67 5.56 scutellospora sp. 1.11 10 s. kowalczyk and j. błaszkowski (koske 1981; schenck, graham and green 1975) and more sandy soils (błaszkowski 1993b). spore density. the overall mean spore density of amf in field soils collected under cultivated plants was 50.7 and ranged from 0 to 925 spores in 100 g dry soil. in the rhizosphere of uncultivated plants, the values were 77.3 and 0 to 865 in 100 g dry soil, respectively. most spores were isolated from under beta vulgaris, hypericum perforatum, polygonum persicaria, and trifolium arvense (tabs 3 and 4). table 2 dominance of amf associated with cultivated (a) and uncultivated (b; not protected) plants of the lubuskie province dominance (%) fungus trap cultures with field soils p. lanceolata z. mays a b a b a b 1 2 3 4 5 6 acaulospora capsicula 0.07 acaulospora lacunosa 0.22 acaulospora koskei 0.01 acaulospora mellea 0.06 acaulospora paulinae 0.01 0.02 ambispora gerdemannii 0.59 0.09 archaeospora trappei 2.33 0.42 1.50 entrophospora baltica 0.01 entrophospora infrequens 0.01 0.46 7.00 0.07 4.13 gigaspora gigantea 0.03 gigaspora margarita 0.06 0.09 0.09 gigaspora sp. 0.06 glomus aggregatum 1.78 2.54 0.08 1.90 0.08 0.06 glomus badium 0.24 12.84 1.90 0.26 glomus caledonium 2.17 0.27 2.47 6.04 8.74 2.34 glomus claroideum 4.10 3.10 67.44 27.37 66.00 56.72 glomus clarum 3.29 glomus constrictum 4.43 32.91 0.17 4.01 0.12 2.17 glomus deserticola 53.47 17.04 6.07 5.53 0.01 glomus fasciculatum 0.20 1.23 0.02 0.22 0.03 glomus fuegianum 2.81 glomus geosporum 0.59 0.25 0.42 glomus lamellosum 0.07 10.43 0.04 13.15 glomus macrocarpum 0.35 4.23 0.02 glomus microcarpum 0.11 4.27 2.86 0.03 glomus mosseae 17.64 3.10 10.59 4.60 15.00 8.49 glomus pansihalos 0.07 glomus pustulatum 0.16 0.49 glomus rubiforme 3.16 1.10 glomus verruculosum 0.01 0.15 glomus 178 0.46 0.75 2.45 0.84 glomus sp. 0.03 0.15 2.11 0.03 0.06 pacispora franciscana 4.67 0.79 3.27 0.84 4.01 0.32 pacispora scintillans 5.90 0.46 2.40 1.24 0.60 0.17 paraglomus laccatum 0.02 4.33 scutellospora armeniaca 0.07 0.04 0.75 scutellospora dipurpurescens 4.30 7.36 6.30 17.23 1.83 2.40 scutellospora pellucida 2.25 0.46 scutellospora sp. 0.22 arbuscular mycorrhizal fungi 11 species density. the overall mean species density of amf in field soils from under cultivated plants was 2.44 and ranged from 0 to 7 in 100 g dry soil. the mean species density of amf associated with uncultivated plant was 2.63 within the range 0-6 in 100 g dry soil. in trap cultures with soils and roots of cultivated plants and the host plants p. lanceolata and z. mays, the overall mean species density was higher by 12.4% and 19.1%, respectively, than in those with soils and roots of uncultivated plants (tabs 3 and 4). in the field, of the cultivated plants, most species were harboured by secale cereale, and of uncultivated plants – cirsium arvense, equisetum arvense and melandrium table 3 spore abundance and species richness of amf associated with roots of 10 cultivated plant species (means) plant species spore abundance species richness field soils* field soils* trap cultures with** p. lanceolata z. mays asparagus officinalis 11.00 2.00 1.78 1.78 avena sativa 46.89 2.56 2.33 2.22 beta vulgaris 119.00 2.56 2.89 2.00 brassica napus 32.25 2.56 1.69 2.22 hordeum vulgare 35.00 2.78 2.56 1.78 secale cereale 54.67 3.22 1.89 2.58 solanum tuberosum 15.11 1.78 2.56 2.20 triticum aestivum 102.78 2.44 2.33 1.56 xtriticum secalum 19.00 1.89 2.50 1.86 zea mays 46.67 2.67 2.78 1.67 * in 100 g dry soil ** in 50 g dry soil table 4 spore abundance and species richness of amf associated with roots of 15 uncultivated (not protected) plant species (means) plant species spore abundance species richness field soils* field soils* trap cultures with** p. lanceolata z. mays achillea millefolium 121.33 3.00 2.33 1.67 carex sylvatica 10.00 2.17 1.67 1.60 cirsium arvense 87.33 4.00 3.50 2.33 corynephorus canescens 6.00 1.17 1.33 0.17 equisetum arvense 67.17 3.67 0.67 2.00 hypericum perforatum 149.00 3.17 1.33 1.83 juncus effusus 10.33 1.33 1.17 1.67 melandrium album 144.50 3.83 2.50 1.67 plantago arenaria 30.00 2.00 1.83 2.50 plantago lanceolata 55.83 2.83 2.67 1.80 polygonum persicaria 181.17 2.33 2.17 1.50 potentilla anserina 12.40 2.17 1.50 0.00 rumex acetosella 34.83 3.00 1.50 1.17 sedum maximum 35.25 2.33 2.00 2.00 trifolium arvense 277.00 2.50 2.50 2.17 * in 100 g dry soil ** in 50 g dry soil 12 s. kowalczyk and j. błaszkowski album (tabs 3 and 4). in trap cultures representing cultivated plants, most species were found when the growing media were soil-root samples from under b. vulgaris and z. mays (tab. 3). when trap cultures contained soils and roots from under uncultivated plants, most species came from those representing ci. arvense (tab. 4). the species most frequently revealed in the spore populations of amf associated with roots of cultivated and uncultivated plants of the lubuskie province, i.e., gl. claroideum, gl. constrictum, gl. deserticola, gl. mosseae and s. dipurpurescens, have many times been found in cultivated and uncultivated sites of different regions of the world (błaszkowski 1993a; jansa et al. 2002). literature data on the sporulation of amf in cultivated versus uncultivated soils are contradictory. as found in this study and that of błaszkowski (1993a), amf produced more spores in uncultivated soils, probably because of the lack of inhibitory influences of agricultural practices. according to oehl et al. (2005), intensive agricultural farming decreases spore production and the numbers of species, especially those from the genus glomus. in contrast, jansa et al. (2002) concluded that some taxa of amf are activated in conditions of agricultural soils. the over 2-fold higher frequency of occurrence of s. dipurpurescens in uncultivated soils probably resulted from high sensitivity of this species to agricultural practices. spores of scutellospora spp. generally are much larger than those of other amf and easier undergo destructions (błaszkowski 2003). arbuscular mycorrhizae. the occurrence of arbuscular mycorrhizae (am) in this group of plants was determined based on 75 root samples. each plant species was represented by three root samples. arbuscules. of cultivated plants, the highest levels of root colonization by arbuscules were found in s. cereale and z. mays (tab. 5). in wild plants, most arbuscules occurred in roots of hy. perforatum, plantago arenaria, and p. lanceolata. no arbuscules were found in roots of the cultivated brassica napus, b. vulgaris and solanum tuberosum and the uncultivated carex sylvatica, e. arvense, and pol. persicaria. vesicles. of the 10 cultivated plant species, only roots of ho. vulgare and z. mays contained a high number of vesicles (tab. 5). of the wild plants, most vesicles were found in roots of p. arenaria, p. lanceolata, and sedum maximum. intraradical hyphae. most intraradical hyphae had roots of the cultivated hordeum vulgare and z. mays and the wild hy. perforatum and sed. maximum (tab. 5). no intraradical hyphae were revealed in roots of b. vulgare, br. napus, c. sylvatica, e. arvense, and so. tuberosum. according to sanders et al. (1977), already a 10% level of root colonization by amf significantly increases the absorption of p from the soil. volkmar and woodbury (1989) found that 2-7% colonization of roots by amf increased up to 25% the shot weight of ho. vulgare. the presence of arbuscules indicates a functional am (smith, read 2008). however, arbuscules were not found in roots of b. vulgaris sampled in this study, which, on the other hand, harboured abundant spore populations of amf. intraradical components of am of many species of the glomeromycota either stain faintly or not at all and, thereby, may be omitted (stutz, morton 1996). in the literature, the amount of data on the active functioning of am in roots of members of the family chenopodiaceae increases (landwehr et al. 2002). arbuscular mycorrhizal fungi 13 arbuscular mycorrhizal fungi and arbuscular mycorrhizae associated with protected plants arbuscular mycorrhizal fungi. the occurrence of amf associated with protected plants was determined based 48 rhizosphere soil-root samples collected under 6 plant species being fully or partly protected (fig. 1). each plant species was represented by 8 soil-root mixtures sampled at four sites of the łagowo landscape park located in łagowo and at two sites of the pszczewo landscape park positioned in trzciel (fig. 1). spores of amf occurred in 31.2% of soil-root samples. they belonged to 11 species of glomus, two species each of acaulospora and scutellospora, one species each of ambispora, archaeospora and pacispora, two undescribed morphotypes (one each of glomus and scutellospora), and not numerous spores of unrecognized fungi forming glomoid spores (tab. 6). amf sporulated in 29 trap cultures with p. lanceolata as the host plant, i.e., 60.4% of all trap cultures established. the spores represented 12 species and two undescribed morphotypes (one each of glomus and scutellospora; tab. 6). frequency of occurrence of species. the species of amf most frequently cooccurring with roots of protected plants (present in >18% of samples and trap cultures) were gl. claroideum, gl. constrictum, and s. dipurpurescens (tab. 6). table 5 mean percent of root length of cultivated and uncultivated (not protected) plants of the lubuskie province with arbuscules, vesicles, and intraradical hyphae of amf plant species arbuscules vesicles hyphae cultivated plants asparagus officinalis 5.00 8.00 25.00 avena sativa 3.00 5.00 46.0 beta vulgaris 0.00 0.00 0.00 brassica napus 0.00 0.00 0.00 hordeum vulgare 18.00 22.00 59.00 secale cereale 32.00 1.00 53.00 solanum tuberosum 0.00 0.00 0.00 triticum aestivum 5.00 9.00 53.00 xtriticum aestivum 3.00 17.00 51.00 zea mays 38.00 20.00 66.00 uncultivated plants achillea millefolium 7.00 20.00 50.00 carex sylvatica 0.00 0.00 0.00 cirsium arvense 19.00 9.00 15.00 corynephorus canescens 5.00 10.00 40.00 equisetum arvense 0.00 0.00 0.00 hypericum perforatum 27.00 21.00 65.00 juncus effusus 5.00 15.00 42.00 melandrium album 1.00 12.00 49.00 sedum maximum 1.00 24.00 66.00 plantago arenaria 28.00 40.00 42.00 plantago lanceolata 31.00 31.00 22.00 polygonum persicaria 0.00 1.00 49.00 potentilla anserina 2.00 18.00 51.00 rumex acetosella 4.00 4.00 22.00 trifolium arvense 1.00 8.00 53.00 14 s. kowalczyk and j. błaszkowski dominance. the eudominants (d>20%) were gl. badium and gl. constrictum (tab. 6). the group of dominants (d=10-20%) formed a. lacunosa and gl. microcarpum. none of the other species found was a subdominant (d=5-10%). spore density. the overall mean spore density of amf in field soils was 7.00 and ranged from 0 to 100 spores in 100 g dry soil. most spores were associated with roots of jovibarba sobolifera, and least with those of lycopodium clavatum (tab. 7). species density. the plant species harbouring most species was j. sobolifera (tab. 7). relatively high numbers of species were also associated with roots of convalaria majalis and l. clavatum. in zubek’s et al. (2005) investigations, the overall mean species richness of amf co-existing with protected plants of the mountain botanical garden in zakopane table 6 frequency of occurrence and dominance of amf associated with roots of protected plant species frequency of occurrence (%) dominance fungus field soils trap cultures with p. lanceolata field soils trap cultures with p. lanceolata acaulospora lacunosa 6.25 17.86 acaulospora mellea 2.08 0.03 ambispora gerdemannii 2.08 0.30 archaeospora trappei 6.25 0.62 glomus aggregatum 2.08 0.03 glomus badium 2.08 29.76 glomus claroideum 4.17 20.83 2.38 79.29 glomus constrictum 18.75 2.08 24.11 0.03 glomus geosporum 2.08 0.30 glomus lamellosum 12.50 5.27 glomus macrocarpum 2.08 0.03 glomus microaggregatum 2.08 2.98 glomus microcarpum 4.17 16.67 glomus mosseae 6.25 6.25 1.49 1.46 glomus verruculosum 2.08 0.03 glomus 178 2.08 0.35 glomus sp. 6.25 6.24 paraglomus laccatum 8.33 2.29 scutellospora dipurpurescens 4.17 20.83 2.68 3.78 scutellospora pellucida 2.08 4.17 1.49 0.49 scutellospora 179 2.08 0.03 table 7 spore abundance and species richness of amf associated with roots of six protected plant species in field soils (a) and trap cultures with p. lanceolata as the host plant (b; means) spore abundance species richness plant species a b a b convallaria majalis 12.5 207.9 0.13 1.1 hedera helix 1.0 19.9 0.50 0.7 helichrysum arenarium 7.2 16.4 0.25 0.7 jovibarba sobolifera 17.5 16.2 1.13 1.2 lycopodium clavatum 0.6 30.2 0.38 1.0 vinca minor 3.1 69.7 0.87 0.6 arbuscular mycorrhizal fungi 15 was higher than that found in this study. however, in both investigations the species most frequently identified were gl. claroideum and gl. constrictum. arbuscular mycorrhizae.the occurrence of am of protected plants of the lubuskie province was determined based on 18 roots samples from six plant species. each plant species was represented by three samples. arbuscules. the highest levels of root colonization by arbuscules were found in con. majalis and helichrysum arenarium (tab. 8). no arbuscules were found in roots of hedera helix and l. clavatum. vesicles. most vesicles were found in roots of hel. arenarium, then in those of j. sobolifera and vinca minor (tab. 8). intraradical hyphae. the plant species harbouring most intraradical hyphae was con. majlis (tab. 8). no intraradical hyphae were observed in roots of h. helix and l. clavatum. the occurrence of spores of amf vs. am vs. in connection with soil chemical properties soil ph ranged from 3.67 to 8.28. the ranges of the total contents (g.kg-1 of dry matter) of organic c, n, p, and k were 0.42-2.0, 0.30-22.5, 0.13-2.02, and 0.10-4.63, respectively. the values of available forms of p, k, and mg (g.kg-1 of dry matter) ranged 9.4-361, 4.90-327, and 13.4-98.8, respectively. rectilinear correlation analyses showed that the spore numbers of amf associated with roots of wild plants significantly correlated with the contents of organic c (r=0.38; p,0.005) and available mg (r=0.55; p<0.05). the contents of total (r=0.50; p<0.005) and available k (r=0.43; p<0.05) in soils from under cultivated plants influenced the abundance of gl. mosseae spores. the contents of available mg in soils of cultivated sites was linked with the numbers of gl. claroideum spores (r=0.90; p<0.05). in uncultivated soils, the contents of organic c and total n and mg correlated with numbers of gl. constrictum spores: r=0.66, p<0.05; r=0.59, p<0.05; and r=0.62, p<0.05, respectively. the levels of mycorrhizal colonization did not correlate with any of the soil chemical properties considered. insignificant also were correlations between numbers of spores and species and levels of mycorrhizal colonization. the results of our study contradict those of many workers who proved that higher contents of soil p generally decrease the production of spores by amf and the composition and distribution of structures of am inside roots (smith, read 1990). alternatively, the activity of amf did not correlate with soil p because of its changing availability at different levels of ph (hayman 1970). table 8 mean percent of root length of protected plants of the lubuskie province with arbuscules, vesicles, and intraradical hyphae of amf plant species arbuscules vesicles hyphae convallaria majalis 29.00 8.00 60.00 hedera helix 0.00 0.00 0.00 helichrysum arenarium 29.00 31.00 50.00 jovibarba sobolifera 19.00 23.00 52.00 lycopodium clavatum 0.00 0.00 0.00 vinca minor 1.00 22.00 54.00 16 s. kowalczyk and j. błaszkowski the lack of relationships between the level of mycorrhizal colonization and the spore abundance and the species richness may have resulted from that the stains used did not reveal structures of all amf in reality existing inside roots (morton, redecker 2001). acknowledgements. this study was supported in part by the polish committee of scientific researches, grants no. 164/n-cost/2008/0 and n n304 061739. references anderson r. c., liberta a. e., dickman l. a. 1984 interaction of vascular plants and vesicular-arbuscular mycorrhizal fungi across a soil moisture-nutrient gradient. oceologia 64: 111–117. bever j., morton j. b., antonovics j., schultz p. a. 1996. host-dependent sporulation and species diversity of arbuscular mycorrhizal fungi in a mown grassland. j. ecol. 84: 71–82. błaszkowski j. 1988. four new species of the endogonaceae (zygomycotina) from poland. karstenia 27: 37–42. błaszkowski j. 1993a. comparative studies of the occurrence of arbuscular fungi and mycorrhizae (glomales) in cultivated and uncultivated soils of poland. acta mycol. 28: 93–140. błaszkowski j. 1993b. the occurrence of arbuscular fungi and mycorrhizae (glomales) in plant communities of maritime dunes and shores of poland. bull. pol. ac. sci. biol. sci. 41: 377–392. błaszkowski j. 2003. arbuscular mycorrhizal fungi (glomeromycota), endogone and complexipes species deposited in the department of plant pathology, university of agriculture in szczecin, poland. http://www.agro.ar.szczecin.pl/~jblaszkowski/. błaszkowski j., kovács g.m., balázs t. 2009. glomus perpusillum, a new arbuscular mycorrhizal fungus. mycologia 101: 247–255. błaszkowski j., renker c., buscot f. 2006. glomus drummondii and g. walkeri, two new species of arbuscular mycorrhizal fungi (glomeromycota). mycol. res. 110: 555–566. błaszkowski j., ryszka p., oehl f., koegel s., wiemken a., kovács g. m., redecker d. 2009. glomus achrum and g. bistratum, two new species of arbuscular mycorrhizal fungi (glomeromycota) found in maritime sand dunes. botany 87: 260–271. błaszkowski j., tadych m., madej m. 2000. glomus minutum, a new species in glomales (zygomycetes) from poland. mycotaxon 76: 187–195. błaszkowski j., tadych m., madej t. 2002. arbuscular mycorrhizal fungi (glomales, zygomycota) of the błędowska desert. acta soc. bot. pol. 71: 71–85. gerdemann j. w. 1968. vesicular-arbuscular mycorrhiza and plant growth. annu. rev. phytopath. 6: 397–418. górny m., gruma l. 1981. metody stosowane w zoologii gleby. pwn, warszawa. grey w. e. 1991. influence of temperature on colonization of spring barleys by vesicular-arbuscular mycorrhizal fungi. plant and soil 137: 181–190. jansa j., mozafar a., anken t., ruh r., sanders i. r., frossard e. 2002. diversity and structure of amf communities as affected by tillage in a temperate soil. mycorrhiza 12: 225–234. kaczorowska z. 1962. opady w polsce w przekroju wieloletnim. prace geogr. ig pan 33:1–107. klironomos j. n., mccune j., hart m., neville j. 2000. the influence of arbuscular mycorrhizae on the relationship between plant diversity and productivity. ecol. lett. 3: 137–141. koske r. e. 1981. a preliminary study of interactions between species of vesicular-arbuscular mycorrhizal fungi in a sand dune. trans. br. mycol. soc. 76: 411–416. koske r. e., gemma j. n., corkidi l., sigüenza c., rinkón e. 2004. arbuscular mycorrhizas in coastal dunes. 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baza (granada, spain). mycologia 100: 296–305. porter w. m., robson a. d., abbott l. k. 1987. field survey of the distribution of vesicular-arbuscular mycorrhizal fungi in relation to soil ph. j. appl. ecol. 24: 659–662. sanders f. e., tinker p. b., black r. l. b., palmersley s. m. 1977. the development of endomycorrhizal root systems. i. spread of infection and growth-promoting effects with four species of vesiculararbuscular endophytes. new phytol. 77: 257–268. schenck n. c., graham s. o., green n. e., 1975. temperature and light effects on contamination and spore germination of vesicular-arbuscular mycorrhizal fungi. mycologia 57: 1189–1194. schönbeck f. 1978. einfluss der endotrophen mykorrhiza auf die krankheitsresistenz höherer pflanzen. z. pflkrankh. pflschutz. 85: 191–196. schüßler a., schwarzott d., walker c. 2001. a new fungal phylum, the glomeromycota: phylogeny and evolution. myc. res. 105: 1413–1421. sieverding e., oehl f. 2006. revision of entrophospora and description of kuklospora and intraspora, two new genera in the arbuscular mycorrhizal glomeromycetes. j. appl. bot. food qual. 80: 69–81. smith s. e., read d. j. 2008. mycorrhizal symbiosis. 3rd ed. academic press: harcourt brace & company, publishers. san diego, london, new york, boston, sydney, tokyo, toronto. stutz j. c., morton j. b. 1996. successive pot cultures reveal high species richness of arbuscular mycorrhizal fungi in arid ecosystems. can. j. bot. 74: 1883–1889. volkmar k. m., woodbury w. 1989. effects of soil temperature and depth on colonization and root and shoot growth of barley inocu lated with vesicular-arbuscular mycorrhizae indigenous to can adian prairie soil. can. j. bot. 67: 1702–1707. walker c., vestberg m., demircik f., stockinger h., saito m., sawaki h., nishmura i., schüßler a. 2007. molecular phylogeny and new taxa in the archaeosporales (glomeromycota): ambispora fennica gen. sp. nov., ambisporaceae fam. nov., and emendation of archaeospora and archaeosporaceae. mycol. res. 111: 137–153. zubek sz., turnau k., błaszkowski j. 2005. arbuscular mycorrhiza of plants from the mountain botanical garden in zakopane. acta mycol. 40 (1): 25–41. 18 s. kowalczyk and j. błaszkowski arbuskularne grzyby mikoryzowe (glomeromycota) związane z korzeniami roślin województwa lubuskiego streszczenie przedstawiono i przedyskutowano wyniki badań występowania arbuskularnych grzybów mikoryzowych (agm) i mikoryz arbuskularnych z gromady glomeromycota związanych z korzeniami 31 gatunków roślin uprawnych, nieuprawnych i chronionych rosnących w 103 stanowiskach województwa lubuskiego. najczęściej znajdywanymi agm byli przedstawiciele rodzaju glomus. często ujawniano również gatunki z rodzaju scutellospora. populacje zarodników agm na ogół były bardziej obfite i różnorodne w glebach uprawnych. większość gatunków roślin chronionych utrzymywała agm. 2014-01-01t11:51:47+0100 polish botanical society new localities of rare species of the genus microbotryum andrzej chlebicki department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, a.chlebicki@botany.pl chlebicki a.: new localities of rare species of the genus microbotryum. acta mycol. 45 (2): 169–173, 2010. microbotryum bistortum, m. dianthorum, m. superbum and m. langerheimii, rare species of smut fungi are reported from new localities in poland, mostly in protected areas. dianthus superbus subsp. superbus is a new host for m. superbum observed. taxonomical status of m. carthusianorum and m. superbum is discussed in the paper. key words: pucciniomycotina, occurrence, ecology, protected areas introduction twenty eight species of the genus microbotryum lév. belonging to microbotryales (pucciniomycotina) have been recorded in poland so far (piątek et al. 2005; majewski et al. 2008; lutz et al. 2008). these fungi are specialized plant pathogens attacking anthers, rarely ovaries, semens and leaves. recent changes in the fungal systematics concern also the genus microbotryum. last investigations show that the species of the genus microbotryum invading plant anthers of caryophyllaceae form a monophyletic group (almaraz et al. 2002, begerow et al. 2004; lutz et al. 2005). in spite of this statement some authors include species invading plants from other families in the genus microbotryum (vánky 1998, 2004; lutz et al 2005). fungi from the genus microbotryum which infect plants from the family caryophyllaceae form sori, mostly in plant anthers, whereas m. major (j. schröt.) g. deml & oberw. form sori also in plant ovaries, anthers and basal part of petals. these fungi are not always host specialized, some of them are restricted to single plant genus, others can infect different plant genera. moreover, the same host plant can be invaded by various anther smuts (see dianthus carthusianorum in this article). revisions of material collected in poland as well as corrections of nomenclature are necessary to keep up with changes in the fungal taxonomy. acta mycologica vol. 45 (2): 169–173 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 170 a. chlebicki distribution of these fungi in poland as well as their host range are poorly known. few localities of the species were reported from some national parks and reserves. new information on four rare species is presented in the paper. methods collected material was examined under light microscope nikon eclipse 91. the spores were mounted in lactophenol and gently heated to boiling point (vánky 1994). for each specimen at least 30 spores were measured. results microbotryum bistortarum (dc.) vánky fig. 1a mycotaxon 67: 40, 1998. basionym: uredo bistortarum dc., in de candolle & lamarck, fl. franç., edn 3 (paris) 5/6: 76, 1815. caeoma bistortarum (dc.) link, willd. sp. plant. 6(2): 10, 1825. ustilago bistortarum (dc.) körn., hedwigia 16: 38, 1877. bauhinus bistortarum (dc.) denchev, mycotaxon 65: 421, 1997. locality. kraków, nowa huta, protected ecological area „łąki nowohuckie”, in flowers of polygonum bistorta l., 21 may 2009, coll. a. chlebicki, kram f-47391 (fig. 1a). comments. so far the fungus has been reported only from areas of karkonosze mts near karpacz (schroeter 1887), mt. śnieżnik, babia góra massif and tatra mts (kochman, majewski 1973). the fungus was noted mainly in flowers of polygonum bistorta, but in the tatra mts it was also observed in bulbils of p. viviparum l. (kochman, majewski 1973). only four attacked plants were noted in the new locality „łąki nowohuckie” in spite of the fact that the host plant occurs very commonly here. microbotryum dianthorum (liro) h. scholz et i. scholz englera 8: 206, 1988. s. lato. basionym: ustilago dianthorum liro, ann. acad. sci. fenn., ser. a 17(1): 35, 1924. locality. warmia region, „rzeka drwęca” reserve, pusta dąbrówka, in the meadow near drwęca river, in anthers of dianthus carthusianorum l., 19 july 2008, coll. a. chlebicki, kram f-46785. sudetes, góry złote mts, meadow near mąkolno and laski, in anthers of dianthus carthusianorum, 12 october 2007, coll. a. chlebicki, kram f-56614. comments. only few specimens of the host plant were noted in the meadows, all of them were invaded by the fungus. according to gac et al. (2007) and refrégier et al. (2008) microbotryum dianthorum is probably a collective species, which occurs on various species of the genus dianthus. recently, denchev et al. (2009) separated a new species from m. dianthorum – m. carthusianorum denchev, giraud & m. e. hood, which occurs on d. carthusianorum. however, on the same host plant another fungus species – microbotryum shykoffianum giraud, denchev & de hood fig. 1. a – microbotryum bistortarum in flowers of polygonum bistorta; b – m. superbum in anthers of dianthus superbus subsp. superbus; c – microbotryum lagerheimii in anthers of lychnis viscaria. new localities of the genus microbotryum 171 can occur. separation of this new species is controversial and not accepted by some mycologists. at this moment the author temporarily included specimens collected on dianthus carthusianorum in microbotryum dianthorum s. lato. an acceptance of the new taxonomical decision will be possible after obtaining of precise descriptions of these two microbotryum species inhabiting dianthus carthusianorum (m. carthusianorum and m. shykoffianum). these species do not differ morphologically, and their identification is possible only on the basis of molecular examination of dna sequences of β-tub, γ-tub and ef1 α. microbotryum superbum (liro) denchev, t. giraud & m. e. hood fig. 1b mycol. balcanica 6: 83, 2009. basionym: ustilago superba liro, ann. acad. sci. fenn., ser. a, 17(1): 37, 1924. locality. podlasie region, pine forest near mikaszewo lake, the area protected in the frame of natura 2000 programme „puszcza augustowska”, in anthers of dianthus superbus l. subsp. superbus, 11 june 2008, coll. a. chlebicki, kram f-46708 (fig. 1b). comments. nearly half of an abundant population of dianthus superbus subsp. superbus was invaded by fungus. specimens inhabiting dianthus superbus were in the past placed in microbotryum violaceum (pers.) g. deml & oberw. (denchev, sharkova 1997). earlier liro (1924) described a new species of fungus on the same host and named it ustilago superba liro. denchev et al. (2009) followed his decision and proposed a new combination – microbotryum superbum (liro) denchev giraud & m. e. hood. refrégier et al. (2008) reported this species as mvdsp.2 in the flowers of d. superbus, d. monspessulanus l. and d. gratianopolitanus vill. microbotryum superbum differs from the m. dianthorum in physiological and genetic characters, but morphology is of their spores identical (le gac et al. 2007; denchev et al. 2009). my observations partially confirm the opinion of these authors (denchev et al. 2009). the host populations of dianthus superbus subps. superbus and d. carthusianorum examined were growing separately and occupied different niches. microbotryum dianthorum s. lato was noted in poland on six species of dianthus (kochman, majewski 1973), among them also on d. superbus subsp. speciosus (rchb.) hayek from the babia góra massif. majewski et al. (2008) listed all known localities of m. dianthorum from poland. d. superbus subsp. superbus is a new host plant for m. superbum in this country. microbotryum lagerheimii denchev fig. 1c mycol. balcanica 4: 64, 2007. locality: western carpathians, beskid sądecki mts, poprad landscape park, meadow on mt. okrąglica południowa, in the anthers of lychnis viscaria l., 1 june 2008, coll. a. chlebicki, kram f-46668 (fig. 1c). comments: host plants were growing abundantly along roads and in meadows (ca some hundred specimens). almost all of them were parasitized by the fungus. previously the fungus was noted on lychnis viscaria (=viscaria vulgaris) in some localities in lower silesia: jarnołtów near wrocław, pątnów legnicki, ratyń near środa śląska, kłodzko, dzierżoniów, bystrzyca górna near świdnica, łęknice near żarów as ustilago antherarum (dc) fr. and u. violacea (pers.) roussel (schroeter 1887; denchev 2007), also in mazowsze region and puszcza kozienicka and na172 a. chlebicki tolin near warszawa, as ustilago violacea and u. silenes-inflatae liro (kochman, majewski 1973; denchev 2007). denchev (2007) separated microbotryum lagerheimii from m. silenes-inflatae (liro) g. deml & oberw. on the basis of colour of ustilospore mass, characters of ustilospores and range of the host plants. so far in poland, specimens parazitizing lychnis viscaria have been included in the species m. silenes-inflatae (majewski et al. 2008). denchev (2007) examined anther smuts collected on this host in poland and transferred them to m. lagerheimii. sori of the specimens collected by the author have pale vinaceous to hazel coloured powdery mass of ustilospores (fig. 1c), typical of m. lagerheimii. acknowledgements. the author thanks dr. marcin piątek and anonymous reviewer for valuable comments, as well as dr. cvetomir denchev for sending his new articles and short discussion of taxonomy of m. dianthorum. the study was supported by the ministry of science and higher education, poland (project no. n n304 328336). references almaraz t., roux c., maumont s., durrieu g. 2002. phylogenetic relationships among smut fungi parasitizing dicotyledons based on its sequence analysis. mycol. res. 106 (5): 541–548. begerow d., göker m., lutz m., stoll m. 2004. on the evolution of smut fungi on their hosts. (in): a. agerer, m. piepenbring, d. blanz (eds). frontiers in basidiomycete mycology. ihw-verlag, eching: 81–98. denchev c., sharkova s. 1997. taxonomic revision of microbotryum (ustilaginales) on dianthus. phytologia balcanica 3 (1): 105–112. denchev c. 2007. microbotryum lagerheimii sp. nov. (microbotryaceae). mycol. balcanica 4: 61–67. denchev c., giraud t., hood m. e. 2009. three new species of anthericolous smut fungi on caryophyllaceae. mycol. balcanica 6: 79–84. kochman j., majewski t. 1973. podstawczaki (basidiomycetes). głowniowe (ustilaginales). (in:) j. kochman, a. skirgiełło (eds). flora polska. rośliny zarodnikowe polski i ziem ościennych. grzyby (mycota), 5. pwn, warszawa-kraków, 270 pp. le gac m., hood m. e., giraud t. 2007. phylogenetic evidence of host-specific cryptic species in the anther smut fungus. evolution 61: 15–26. liro j. i. 1924. die ustilagineen finlands. i. annales academiae scientiarum fennicae, ser a 17(1): i-xviii + 1–636. lutz m., göker m., piątek m., kemler m., begerow d., oberwinkler f. 2005. anther smuts of caryophyllaceae: molecular characters indicate host-dependent species delimitation. mycol. progress. 4: 225–238. lutz m., piątek m., kemler m., chlebicki a., oberwinkler f. 2008. anther smuts of caryophyllaceae: molecular analyses reveal further new species. mycol. res. 112(11): 1280–1296. majewski t., piątek m., ruszkiewicz-michalska m. 2008. ustilaginales. (in:) w. mułenko, t. majewski, m. ruszkiewicz michalska (eds). a preliminary checklist of micromycetes in poland. (in:) z. mirek (ed.). biodiversity of poland 9: 297–318. piątek m., ruszkiewicz-michalska m., mułenko w. 2005. catalogue of polish smut fungi, with notes on four species of anthracoidea. polish bot. j. 50 (1): 19–37. refrégier g., le gac m., jabbour f., widmer a., yockteng r., shykoff j. a., hood m. e., giraud t. 2008. cophylogeny of the anther smut fungi and their caryophyllaceous hosts: prevalence of host shifts and importance of delimiting parasite species for inferring cospeciation. bmc evolutionary biology 8: 100. schroeter j. 1887. die pilze schlesiens: ustilaginei. (in:) f. cohn (ed.). cryptogamen-flora von schlesien 3(1): 261–290. j. u. kern’s verlag, breslau. vánky k. 1994. european smut fungi. g. fischer verlag, stuttgart-new york, 570 pp. vánky k. 1998. the genus microbotryum (smut fungi). mycotaxon 67: 33–60. vánky k. 2004. anther smuts of caryophyllaceae. taxonomy, nomenclature, problem in species delimitation. mycol. balcanica 1: 189–191. new localities of the genus microbotryum 173 nowe stanowiska rzadkich gatunków grzybów z rodzaju microbotryum streszczenie grzyby z rodzaju microbotryum lév. są obecnie zaliczane do podgromady pucciniomycotina, gdzie wchodzą w skład rzędu microbotryales r. bauer & oberw. w polsce dotychczas odnotowano 28 gatunków z tego rodzaju. są to wyspecjalizowane patogeny roślin atakujące głównie pylniki, rzadziej słupki, nasiona i liście. zachodzące w ostatnich latach zmiany w taksonomii wymagają dostosowania nazewnictwa i przejrzenia dotychczas zebranych materiałów. rozmieszczenie tych grzybów w polsce jest słabo poznane. nieliczne stanowiska wielu gatunków tych grzybów były podawane z niektórych parków narodowych i rezerwatów przyrody. również zakres żywicieli jest w polsce niedostatecznie poznany. w niniejszej notatce zostały przedstawione informacje o czterech gatunkach tych grzybów: microbotryum bistortarum, m. dianthorum, m. superbum i m. lagerheimii. 2014-01-01t11:51:14+0100 polish botanical society impact of selected antagonistic fungi on fusarium species – toxigenic cereal pathogens delfina popiel1, hanna kwaśna2, jerzy chełkowski1, łukasz stępień1 and magdalena laskowska3 1institute of plant genetics, polish academy of sciences, strzeszyńska 34, pl-60-479 poznań, jche@igr.poznan.pl 2department of forest pathology, agricultural university, wojska polskiego 71c, pl-60-625 poznań 3department of chemistry, agricultural university wojska polskiego 75, pl-60-625 poznań popiel d., kwaśna h., chełkowski j., stępień ł., laskowska m.: impact of selected antagonistic fungi on fusarium species – toxigenic cereal pathogens. acta mycol. 43 (1): 29–40, 2008. fusarium-ear blight is a destructive disease in various cereal-growing regions and leads to significant yield and quality losses for farmers and to contamination of cereal grains with mycotoxins, mainly deoxynivalenol and derivatives, zearalenone and moniliformin. fusarium pathogens grow well and produce significant inoculum on crop resiudues. reduction of mycotoxins production and pathogen sporulation may be influenced by saprophytic fungi, exhibiting antagonistic effect. dual culture bioassays were used to examine the impact of 92 isolates (belonging to 29 fungal species) against three toxigenic species, i.e. fusarium avenaceum (corda) saccardo, f. culmorum (w.g.smith) saccardo and f. graminearum schwabe. both f.culmorum and f. graminearum isolates produce trichothecene mycotoxins and mycohormone zearalenone and are considered to be the most important cereal pathogens worldwide. infection with those pathogens leads to accumulation of mycotoxins: deoxynivalenol (don) and zearalenone (zea) in grains. fusarium avenaceum isolates are producers of moniliformin (mon) and enniatins. isolates of trichoderma sp. were found to be the most effective ones to control the growth of examined fusarium species. the response of fusarium isolates to antagonistic activity of trichoderma isolates varied and also the isolates of trichoderma differed in their antagonistic activity against fusarium isolates. the production of mon by two isolates of f. avenaceum in dual culture on rice was reduced by 95% to 100% by t. atroviride isolate an 35. the same antagonist reduced the amount of moniliformin from 100 μg/g to 6.5 μg/g when inoculated to rice culture contaminated with mon, which suggests the possible decomposition of this mycotoxin. key words: antagonistic fungi, fusarium, moniliformin, trichoderma acta mycologica vol. 43 (1): 29–40 2008 30 d. popiel et al. introduction fusarium species are cosmopolitan necrotrophic pathogens of cereals, pulse crops and many other plants, important in agricultural and forest landscape. fusa­ rium ear (head) blight (fhb=scab) has been known for more than 100 years and is one of the important diseases of wheat and other small grain cereals. in europe it is caused mostly by f. graminearum, f. culmorum, f. avenaceum and f. poae. other fusarium species are less important due to their lower incidence and aggressiveness (arseniuk et al. 1999; bai, shaner 1994; parry et al. 1995; chełkowski 1998; wakuliński, chełkowski 1993; bottalico, perrone 2002). fhb leads to significant loss of grain yield and quality. several fungal secondary metabolites, e.g. deoxynivalenol (don), nivalenol (niv), zearalenone (zea) and moniliformin (mon) and also their derivatives may contaminate cereal grains (mcmullen et al. 1997; jones, mirocha 1999; bottalico 1998; chełkowski 1998). the disease has been re-emerging in many cereal-growing regions worldwide (parry et al. 1995; mcmullen et al. 1997; jones, mirocha 1999; bottalico, perrone 2002). three chemotypes are common within f. culmorum and f. graminearum species: nivalenol (niv), 3acetyl-deoxynivalenol (3-acdon) and 15acetyl-deoxynivalenol (15-acdon). these chemotypes can be identified by chemical analyses of fungal cultures or by chemotype-specific dna markers (quarta et al.2005). fusarium culmorum and f. graminearum survive saprophytically on plant resi-aprophytically on plant resion plant residues after harvest and serve as a source of inoculum for the subsequent year. both species colonize particularly frequently maize stalks and f. graminearum produces significant amounts of ascospores and conidia (sutton 1982). fusarium avenaceum is a cosmopolitan species and one of the most important ones in agriculture and the forest environment in the moderate climatic zone (nelson et al. 1983; kwaśna et al. 1991; leslie, summerell 2006). simultaneous cooccurrence of the three mycotoxins: monilformin, deoxynivalenol and nivalenol was found in high percentage of positive samples (33% and 35%), during 1998 and 1999 epidemics of fhb in wheat in poland (tomczak et al. 2002). there are few species among soil microorganisms antagonistic to fusarium, able to reduce their population in soil and debris. several fungal species were examined for the ability to reduce the inoculum potential of fusarium pathogens, mainly by reduction of biomass in plant residues colonized by fusarium (łacicowa, pięta 1985; ligitt et al. 1997; luongo et al. 2005; lutz et al. 2003; dawson et al. 2004). several antagonists were found to reduce infection of ears and accumulation of don in wheat grains (ligitt et al. 1997; dawson et al. 2004). clonostachys, gliocladium and trichoderma species reduced the colonization of wheat and maize by pathogenic fusarium species and suppressed the sporulation of the latter (woo et al. 2005; luongo et al. 2005). there is only scarce information on fungal antagonists of toxigenic fusarium species and their ability to reduce the production of mycotoxins in solid substrates. trichoderma species have been examined for more than 50 years and are known as highly effective in biological control of a wide range of plant pathogens of soil origin. they are also known to produce over 120 secondary metabolites, in-they are also known to produce over 120 secondary metabolites, including antifungal metabolites (kubicek, harman 1998; woo et al. 2005). cooney et al. (2001) showed that 6-pentyl-alpha-pyrone (6pap), which is a metabolite of t. harzianum (thf2/3), can reduce the production of deoxynivalenol impact of selected fungi 31 by f. graminearum on agar medium by 66 to 81%. the authors developed an agar bioassay technique to examine fusarium­trichoderma interaction on their secondary metabolites level. t. harzianum type 4 and named t. aggressivum has been found recently to be com-has been found recently to be competitor of commercial mushroom agaricus bisporus, known to be the cause of green mold and significant losses to mahrooms producers (savoie, mata 2003). the aim of this paper was to examine the effect of 29 fungal species isolates (including trichoderma) originated from soil, compost and cereals, on the growth of three toxigenic fusarium species in dual cultures and a mutual interaction between the antagonists and fusarium isolates in bioassays, and to examine the reduction of moniliformin production in dual cultures of f. avenaceum with selected effective trichoderma antagonists. material and methods fungal isolates. isolates of tested fungi originated from culture collections of the institute of plant genetics, polish academy of sciences, poznań, poland (kf and an), the institute of science of food production, bari, italy (item) and the department of forest pathology, agricultural university, poznań, poland (tab. 1). fungal isolates originating from soil, compost and cereal grains were maintained on a synthetic low nutrient agar sna (nirenberg 1981; kwaśna et al.1991). identification and nomenclature of trichoderma and gliocladium isolates was followed according to gams and bissett 1998. fusarium isolates – species and chemotype identification. species were identified based on their macroconidia structure using an olympus optical microscope at a 400-500x magnification according to nelson et al. (1983), kwaśna et al. (1991) and nirenberg (1981) manuals. species identification was also performed using dna markers. fungal dna was extracted using the modified ctab method (chełkowski et al. 2002). pcr amplification mixture consisted of 0.5 u of taq dna polymerase (finnzymes), 2.5 μl of pcr buffer, 12.5 pmol of forward/reverse primers, 2.5 mm of each dntp and about 10 ng of fungal dna. for the identification of f. culmorum, f. graminearum and f. avenaceum species, the following markers were used: fc01 (570 bp), ubc85 (332 bp), fg16n (280 bp), fa (920 bp) (nicholson et al. 1998; schilling et al. 1996, doohan et al. 1998) and to identify 3ac-don, 15ac-don and niv chemotypes tri3 (708 and 354 bp) and tri7 (625 bp) markers were used (quarta et al. 2005). dual culture bioassays. a modified bioassay of mańka (1974) was applied to examine growth reduction of three toxigenic species isolates: f. avenaceum, f. cul­ morum and f. graminearum. a fusarium isolate and a tested fungus (an antagonist) were inoculated onto 9 cm petri dishes at a distance of 1cm on potato dextrose agar (pda). as a control each fungus was cultured separately. agar plates were then incubated at 25˚c at diffused daylight. inhibition of fusarium growth by the antagonist was evaluated using a modified mańka (1974) scale –8 to +8 presented in figure 1, where 0 indicated no inhibition and +8 a total inhibition, with the fusarium mycelium in >95% overgrown by the antagonist. growth rate of each control culture was measured after 2-7 days. at least two replications were performed. 32 d. popiel et al. to examine the reduction of fusarium toxin production in solid substrates 50 g of commercial rice were soaked with 15 ml of distilled water in a 300 ml erlenmayer flask overnight before sterilization run for 30 min. at 121˚c. then it was inoculated with fusarium and the tested fungus (antagonist). inoculation was made with four 4mm diameter discs of each fungus cut from the edge of 1-week-old culture on pda and 2 ml of sterile water. cultures were incubated at 25˚c and shaken each day to support a uniform growth of mycelium. in the control each fungus was cultured ta b l e 1 growth rate of 92 candidate antagonistic isolates from soil, compost and cereals and fusarium isolates on pda medium in mm/day at 25˚c species group species of potential competitors no. of isolates accession code (an) mean growth rate mm/day i trichothecium roseum (pers; fries) link 1 27 10 ii trichoderma viride pers. & fries 12 14, 15, 16, 17, 18, 45, 46, 47, 48, 51, 52, 61 11-21 iii trichoderma pseudokoningii rifai 1 60 21 iv trichoderma polysporum link: pers 1 55 20 v trichoderma longibrachiatum rifai 1 22 21 vi trichoderma koningii oudemans 4 49, 59, 65, 66 21 vii trichoderma harzianum rifai 12 3, 4, 5, 6, 13, 53, 54, 58, 62, 63, 64, 94 21-31 viii trichoderma hamatum (bon) bainier 2 21, 56 20-21 ix trichoderma citrinoviride bissett 1 89 31 x trichoderma aureoviride rifai 1 57 21 xi trichoderma atroviride karsten 4 19, 35, 50, 90 21-30 xii trichoderma asperellum samuels, lieckf. & nirenberg 1 93 25 xiii pythium sp. 1 72 7 xiv paecilomyces carneus (duché & r. heim) a.h.s. br. & g. sm. 1 42 1 xv paecilomyces farinosus holmskjold 1 41 1 xvi mortierella sp. 1 71 14 xvii melanospora fimicola e.c. hansen 3 29, 31, 44 6-21 xx idriella bollei sprague 1 83 18 xviii hypocrea hunua dingley 1 20 21 xix gliocladium virens j.h. mill., giddens & a.a. foster 6 68, 69, 70, 73, 74, 75 20-21 xxi gliocladium roseum bainier 4 24, 25, 26, 78 2-7 xxii gliocladium catenulatum glim & abbot 4 23, 32, 43, 88 7-9 xxiii gilmaniella sp. 4 67, 76, 77, 79 5-7 xxiv fusarium flocciferum corda 3 84, 85, 86 7-13 xxv fusarium equseti (corda) sacc. 7 1, 2, 7, 9, 10, 11, 12 5-15 xxvi epicoccum nigrum link 6 36, 37, 38, 39, 40, 87 2-12 xxvii cladosporium cladosporioides (fr.) de vries 2 81, 82 17-19 xxviii chaetomium cochlioides palliser 4 28, 30, 33, 34 12-21 xxix acremonium sp. 1 80 5 fusarium sp. xxx fusarium avenaceum (corda) sacc. 2 15 xxxi fusarium culmorum (w.g.smith) sacc. 3 19 xxxii fusarium graminearum schwabe 13 9-20 34 d. popiel et al. ranged from 9 to 20 mm/day (tab. 1). three chemotypes were identified using specific primers for pcr amplification of dna among f. culmorum and f. graminear­ um isolates used in our experiments: 3-acdon chemotypes (mostly f. culmorum fig. 2 a. a comparison of impact of 92 candidate antagonistic fungi isolates against two f. graminearum isolates kf 844 and kf 2870 in bioassay on pda medium after 7 days at 25˚c (in –8 to +8 scale of inhibition). fig. 2 b continued. a comparison of impact of 92 candidate antagonistic fungi isolates against two f. graminearum isolates kf 844 and kf 2870 in bioassay on pda medium after 7 days at 25˚c (in –8 to +8 scale of inhibition). impact of selected fungi 35 isolates), 15acdon chemotype (most of f. graminearum isolates) and niv chemotype (several isolates of both species). competitive abilities of candidate fungi against toxigenic fusarium isolates were examined in dual culture bioassays on agar and solid substrate bioassay. fusa­ rium avenaceum, f. culmorum and f. graminearum produce aurofusarin and other carmine-red pigments on pda medium (vesonder, goliński 1989). trichoderma isolates produce a green pigment of polyphenolic nature. colony pigmentation is a useful marker when studying the interaction between fusarium isolates and their antagonists. fusarium isolates growth in dual cultures with competitive isolates were visually reduced after 4 days. a week after inoculation growth inhibition and mycoparasitism of trichoderma isolates were observed – the plate was green and overgrown with trichoderma with significant production of abundant conidia in pustules over mycelium of fusarium isolates, with only a small spot of the plate remaining red. fusarium growth and red pigment production were strongly inhibited by isolates of t. atroviride, t. harzianum, t. hamatum, t. longibrachiatum and t. koningii. isolates of f. graminearum (kf 844 and kf 2870) were grown simultaneously with 92 antagonistic isolates and exhibited different susceptibility to the presence of the antagonists: isolate kf 2870 was significantly less resistant and isolate kf 844 exhibited a higher level of resistance against the same antagonistic mycoparasite (fig. 2a, b). the antibiosis effect was not found in any of the candidate antagonists in dual culture on pda. the interaction between eight isolates of trichoderma and six isolates of fusa­ rium is presented in tables 2 and 3. thus, on one hand the same trichoderma isolate exhibited different antagonism against various fusarium isolates, and on the other hand antagonistic isolates exhibited various aggressiveness against the same isolate of fusarium. trichoderma isolates are able to produce antibiotics – inhibitors of other fungi (such as 6pap), and several enzymes, which hydrolyse fungal structures – conidia and mycelia – and macromolecules – chitin, cellulose, hemicellulose, beta glucan, xylem and proteins (łacicowa, pięta 1985; kubicek, harman 1998; cooney et al. 2001). this ability allows trichoderma isolates to utilise the mycelium of fusarium ta b l e 2 growth inhibition of four isolates of f. graminearum and f. culmorum, belonging to three chemotypes (3acdon, 15acdon and niv) by eight isolates of trichoderma in dual cultures on pda medium after 7 days at 25˚c (in -8 to +8 scale) accession code antagonist species fusarium species and their chemotypes kf 350 f. culmorum niv kf 846 f. culmorum 3ac-don kf 844 f. graminearum 15ac-don kf 2870 f. graminearum 15ac-don an 13 t. harzianum +4 +6 +8 +8 an 16 t. atroviride +5 +6 +5 +6 an 35 t. atroviride +7 +8 +8 +8 an 89 t. citrinoviride +2 +2 +8 +8 an 90 t. atroviride +4 +6 +8 +8 an 92 t. harzianum -2 0 -2 +6 an 93 t. viride +8 +4 +6 +6 an 94 t. harzianum +2 +2 +2 +4 36 d. popiel et al. as a source of nutrients. our biotest experiments confirmed the mycoparasitism of trichoderma species over all three species and chemotypes of toxigenic fusarium. three types of interaction between trichoderma species and plant pathogens have been recognized: antibiosis, competition for nutrients and hyperparasitism (woo et al. 2006). no antibiosis was observed in our experiments. results of our previous paper (buśko et al.2008) correspond well with the finding of cooney et al. (2001) on the impact of t. harzianum isolates on a don-producing f. graminearum in the agar medium bioassay. the same authors proved the inhibition of don production by a trichoderma metabolite 6pap, by as much as 80%. the mechanism of don content reduction in f. graminearum cultures remains unsolved. it was shown that don when added to an agar medium inoculated with trichoderma was not metabolised by the fungus (cooney et al. 2001). a question arises whether isolates of trichoderma, growing on the mycelium of toxigenic fusarium species, are also able to transform or degrade such mycotoxins as don, niv, mon, zea, and others – in total 19 mycotoxins identified in grain samples (chełkowski 1998; bottalico 2002). until now, the ability to decompose don has been found very rarely among microorganisms. only one mixed culture among 1285 microbial cultures, isolated from farmland soil, cereal grains and others sources, transformed don into two products that can be separated chromatographically (mainly 3-keto-4-deoxynivalenol; voelkl et al. 2004). in our previuos laboratory experiments f. culmorum and f. graminearum cultures grown on rice produced five trichothecenes: fusarenone x (up to 21 mg/kg), nivalenol (up to 3.7 mg/kg), deoxynivalenol (up to 310 mg/kg), 3ac-don (up to 228 mg/ kg) and 15ac-don (up to 184 mg/kg). production of five trichothecene mycotoxins don, 3acdon, 15acdon, nivalenol and fusarenone x was reduced by over 95% in dual culture bioassay by trichoderma isolates an 22 and an 35 (buśko et al. 2008). don concentration in the bioassay in microcosms was reduced by 45% and fungal biomass was reduced by 15% in studies of noef et al. 2006. consequently, mean don production per biomass was significantly lower 36% in dual culture with t. atroviride than in solitary culture of f. graminearum and the above mentioned authors did not find any evidence for the role of don production in f. graminearum defence against t. atroviride. ta b l e 3 growth inhibition of two isolates of fusarium avenaceum in dual culture by eight isolates trichoderma (in -8 to +8 scale) on pda medium after 7 days at 25˚c accession code antagonist species fusarium avenaceum isolate kf 203 (atcc 64451) kf 2818 an 13 t. harzianum +4 +4 an 16 t. atroviride +5 +4 an 35 t. atroviride +6 +7 an 89 t. citrinoviride +6 +7 an 90 t. atroviride +7 +8 an 92 t. harzianum +6 +4 an 93 t. viride +2 +2 an 94 t. harzianum +6 +6 impact of selected fungi 37 a significant reduction of wheat head infection by f. graminearum and don accumulation in kernels was found by dawson et al. (2004) when ears were preinoculated by fungal antagonists, such as t. harzianum, clonostachys rosea and f. equiseti. recently three major compounds exhibiting antifungal activity were identified to be produced by t22 and t39 isolates of t. harzianum, that are already used as active agents in a variety of commercial biopesticides (vinale et al. 2006). over 50 commercial biopesticides were found effective in biological control of soil and residue borne pathogens (woo et al. 2005). fusarium avenaceum isolates produced moniliformin up to 100 mg/kg (tab. 4). the isolate t. atroviride an35 was found the most effective among the examined accessions against toxigenic fusarium isolates used in this study. this antagonist was able to reduce moniliformin production in dual culture bioassay on rice by 95-100%, depending on the applied f. avenaceum isolate. the same isolate reduced by 93% the amount of mon in a rice culture of f. avenaceum item 3411 (kf 2603) – from 100 μg/g to 6.5 μg/g (tab. 4). the application of trichoderma competitors may reduce growth of fusarium species through competition in crop debris and mycotoxin production as well. however, the development of the formulation of biofungicides for practical control of diseases remains a very important task (kubicek, harman 1998; woo et al. 2006). both in the literature and in our experiments presented in this paper significant interaction between toxigenic fusarium species and trichoderma competitotors was found to be of complex character, with importance of several characters such as growth rate, enzymes and secondary metabolites production (harman 2006). it can be concluded that competitive trichoderma isolates are candidate fungi for biological control of toxigenic fusarium species aggressive to cereals (such as f. culmorum, f. graminearum and f. avenaceum) and in reducing their inoculum, as well as preventing mycotoxin accumulation in plant tissues and crop residues in field. it seems to be important to underline, that saprophytic species trichoderma har­ zianum and t. atroviride are not antagonistic to a.bisporus. both species isolates can be distinguished by dna analyses from t. aggressivum, however when high amount of inoculum is present in mushroom compost both species may compete for nutrients with a.bisporus mycelium. identification and nomenclature of trichoderma species was recently modified by gams and bissett (1998). there is high similarity ta b l e 4 reduction of moniliformin produced by two isolates f. avenaceum in dual culture bioassay and decomposition of mon by trichoderma harzianum an 35 isolate on rice after 21 days at 25˚c f. avenaceum isolate mon produced by f. avenaceum (μg/g) mon produced in dual culture f. avenaceum/t. atroviride (μg/g) reduction of mon level (%) kf 203 (atcc 64451) 58.5 3.4 94.2 kf 2818 296.2 nd 100 f. avenaceum isolate mon (μg/g) mon after t. atroviride culturing (μg/g) decomposition efficiency (%) kf 2603 (item 3411) 100 6.5 93.5 38 d. popiel et al. of trichoderma isolates morphology under laboratory conditions and there are many confusions in the literature, concerning identification of species. actual list of over 40 species and their descriptions are available on website: http://nt.ars-grin.gov/ taxadescriptions/keys/trichodermaindex.cfm. acknowledgement. this study was partly supported by project no. pbz-kbn-112/p06/2005 of the polish ministry of science and higher education. references arseniuk e., foremska e., góral t., chełkowski j. 1999. fusarium head blight reactions and accumulation of deoxynivalenol (don) and some of its derivatives in kernels of wheat, triticale and rye. j. phytopathol. 147: 577–590. bai g., shaner g. 1994. scab of wheat: prospects for control. plant. dis. reporter 78: 760–765. bottalico a., perrone g. 2002. toxigenic fusarium species and mycotoxins associated with head blight in small-grain cereals in europe. j. pl. pathol. 108: 998–1003. bottalico a. 1998. fusarium diseases of cereals, species complex and related mycotoxin profiles in europe. j. pl. pathol. 80: 85–103. buśko m., chełkowski j., popiel d., perkowski j. 2008. solid substrate bioassay to evaluate impact of trichoderma on trichothecene mycotoxin production by fusarium species. j. sci. food agric. 88: 533–541. chełkowski j. 1998. distribution of fusarium species and their mycotoxins in cereal grains. 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(in:) j. chełkowski (ed.). fusarium, mycotoxins, taxonomy and pathogenicity. elsevier scientific publishers, amsterdam: 39 pp. vinale f., marra r., scala f., ghisalberti el., lorito m., sivasithamparam k. 2006. major secondary metabolites produced by two commercial trichoderma strains active against different phytopathogens. lett. app. microbiol. 43: 143–148. voelkl a., vogler b., schollenberger m., karlovsky p. 2004. microbial detoxifi cation of mycotoxin de-microbial detoxification of mycotoxin deoxynivalenol. j. basic microbiol. 44: 147–156. wakuliński w., chełkowski j. 1993. fusarium species causing scab of wheat, rye and triticale in poland. hod. rośl. aklim. nasien. 37:137–142. woo s. l., scala f., ruocco m., lorito m. 2006 the molecular biology of interactions between tricho­ derma sp., phytopathogenic fungi and plants. phytopathology 96: 181–185. 40 d. popiel et al. antagonistyczne oddziaływanie wybranych grzybów na toksynotwórcze gatunki fusarium patogeniczne dla zbóż s t r e s z c z e n i e fuzarioza kłosa jest w wielu regionach produkujących zboża chorobą wyniszczającą i powoduje straty powstające na skutek obniżenia plonowania i jakości ziarna. ziarno z roślin porażonych jest zanieczyszczone mikotoksynami, przede wszystkim deoksyniwalenolem i jego pochodnymi, zearalenonem i moniliforminą. gatunki fusarium dobrze rozwijają się na resztkach pożniwnych i obficie na nich zarodnikują. grzyby saprotroficzne o cechach antagonistycznych wobec tych patogenów mogą przyczyniać się do zmniejszenia zarodnikowania patogenów fusarium i obniżenia ilości tworzonych przez nie mikotoksyn. antagonistyczne oddziaływanie 92 izolatów grzybów należących do 29 gatunków testowano w bikulturach z izolatami trzech toksynotwórczych gatunków f. avenaceum (corda) saccardo, f. culmorum (w.g.smith) saccardo i f. graminearum schwabe. gatunki f. culmorum i f. graminearum tworzą mikotoksyny trichotecenowe i mikohormon zearalenon oraz należą do najistotniejszych patogenów zbóż w skali światowej. porażenie kłosów zbóż przez te gatunki powoduje akumulację w ziarniakach deoksyniwalenolu (don) i zearalenonu (zea). izolaty f. avenaceum tworzą moniliforminę (mon) i enniatyny. izolaty gatunków trichoderma okazały się najbardziej efektywnymi dla redukcji wzrostu izolatów wymienionych gatunków. efekt antagonistyczny poszczególnych izolatów trichoder­ ma względem tych samych izolatów fusarium różnił się znacząco. również stopień redukcji wzrostu poszczególnych izolatów fusarium przez te same izolaty trichoderma był znacząco różny. ilość moniliforminy produkowanej przez dwa izolaty f. avenaceum w bikulturach na ryżu była redukowana o 95-100% przez izolat t. atroviride an35. ten sam grzyb antagonistyczny redukował zawartość moniliforminy z poziomu 100 μg/g do 6.5 μg/g w kulturze na ryżu, co sugeruje możliwość dekompozycji tej mikotoksyny przez ten izolat. 2014-01-01t11:47:03+0100 polish botanical society revised data on the occurrence of myxomycetes in central poland maria ławrynowicz1, dominika ślusarczyk1 and agnieszka salamaga2 1department of mycology, university of łódź banacha 12/16, pl-90-237 łódź, miklaw@biol.uni.lodz.pl 2institute of botany, jagiellonian university, kopernika 27 pl-31-501 kraków, asalamaga@wp.pl ławrynowicz m., ślusarczyk d., salamaga a.: revised data on the occurrence of myxomycetes in central poland. acta mycol. 46 (2): 223–232, 2011. a checklist of 81 taxa has been prepared on the basis of 750 exsiccates of slime moulds collected in the years 1958-2009 in the area of central poland and preserved in the herbarium lod f. all these materials were reexamined according to contemporary literature. the paper summarizes the existing data concerning the occurrence of slime moulds in this area. among the identified taxa there are four species indicated in the red list in poland: badhamia affinis, physarum robustum, didymium leptotrichum, and clastoderma debaryanum. key words: slime moulds, checklist, biodiversity, distribution introduction slime moulds (myxomycetes, mycetozoa) are the group of organisms of increasing importance in biological and phylogenetical respects. their morphological, ecological and genetic diversity is the matter of interest for specialists worldwide (e.g., ing 1994; heilmann-clausen 2001; krieglsteiner 2004). in poland, a vast programme of biodiversity studies has been recently undertaken (mirek ed. 2002-2009) resulting, among other works, in a checklist of 222 myxomycetes, that includes all hitherto published taxa (drozdowicz, ronikier, stojanowska and panek 2003). recently, a field study in ne poland has brought an interesting and rich collection of slime moulds (panek, romański 2010). the above publications stimulated the authors of the present paper to examine and revise slime moulds collections from the area of central poland, preserved in the herbarium universitatis lodziensis (lod f). until now, some data concerning slime moulds in central poland were published by orzechowski (1966) – from the łódź city and adjacent areas, kalinowska-kucharska (1975) – from several localities in the area including all the myxomycetes collected in the nature reserves in course of mycocoenological studies by ławrynowicz (1973). at present, a detailed study of slime moulds has acta mycologica vol. 46 (2): 223–232 2011 revised data on the occurrence of myxomycetes in central poland 225 a watershed between the catchment areas of warta and vistula rivers, running from the south to the north and dividing the area into two parts. as a consequence of geological, climatic and hydrological characters, the northern occurrence limits of three important forest trees: fagus sylvatica, picea abies and abies alba run through this area. the most valuable stands of these tree species are protected in nature reserves. transitional character of central poland creates specific conditions for vegetation cover and communities of different organisms, including slime moulds. the materials studied come from 34 localities indicated on the map (fig. 1). material and methods in total 750 records of slime mould collected in the period 1958-2009 are the subject of interpretation. the specimens were analyzed using routine microscopic and laboratory techniques. the identification of species was carried out according to specialistic literature, e.g.: krzemieniewska (1960), neubert, nowotny and baumann (1993, 1995), neubert et al. (2000), martin & alexopoulus (1969), ing (1999) and nannenga-bremekamp (1991). the nomenclature follows drozdowicz et al. (2003), updated according to lado (2005-2011). table 1 correspondence between sources of information (tab.2) and study areas 1 2 3 4 d ba b k d g kw js k l g kw l i pk m pj m msz nw w mur nw o o pj pk pk s s t w t ii ─ ─ ─ arr bruż bruż gr jas gr j jas iii kr j pap ka kr luć ka lł luć rd p p pap sz rd sz lubl lubl iv nak ─ ─ nak plz plz the columns 1-4 correspond to the sources of information 1-4 included in table 2. each of them is divided into four parts corresponding to the four types of study areas: i – nature reserve, ii – the arboretum, iii – forests, iv – nature sites. 226 m. ławrynowicz et al. the detailed register prepared in course of analyzes of all exsiccates (ławrynowicz, ślusarczyk and salamaga 2010) was a basis of synthesis presented in table 2. the arrangement of taxa is adopted according to śliwa (2010) and modified for slime moulds. the sources of data (tab. 2, column 3) are two types: published data (1, 2, 3) and unpublished data 4 – (numbers of herbarium exsiccates). the specimens considered were collected with different intensity and different methods; great part of them were recorded during thorough mycocoenological or other investigations focused on fungi, but some were just single, accidental findings. the data represent four types of study areas: nature reserves, arboretum, forests, and nature sites (tab. 1). all herbarium vouchers are preserved in herbarium universitatis lodziensis (lod f). results and discussion macroand micromorphological analysis of exsiccates of slime moulds preserved in the herbarium of the łódź university (lod f) resulted in the identification of 81 taxa (78 species and 3 varieties). the collection examined contains the records gathered in central poland before 2010. the data are summarized in a checklist (tab. 2). although the list covers only a number of collecting places and does not provide full information on the occurrence and distribution of slime moulds in the area concerned, it gives several interesting data. among the slime moulds collected there is a group of widely distributed and frequently noted species: lycogala epidendrum, fuligo septica, ceratiomyxa fruticulosa. there is also a group of rare slime moulds, among them of redlisted species: badhamia affinis, physarum robustum, didymium leptotrichum, and clastoderma debaryanum (drozdowicz, ronikier and stojanowska 2006). the material examined is not a sufficient basis to describe any species as rare in a broader sense, especially in the case of the taxa observed accidentally at a single locality, but known to be common in other regions. this preliminary checklist indicates that slime moulds can colonize a variety of sites from nature reserves to anthropogenic places. the first collections of slime moulds in the area of łódź city and its surroundings (1958-1963) were made by orzechowski (1966) and reveal the presence of rare species in city parks and small forests. the paper by kalinowska-kucharska (1975) indicates that nature reserves are the places hosting the greatest variety of slime moulds. the materials presented in this work were collected during mycocoenological studies in permanent plots in the tilio-carpinetum and potentillo albae-quercetum associations, in the area of 7 nature reserves: ostrowy, molenda, nowa wieś, komasówka, dębowiec, and trębaczew. other mycocoenological investigations including myxomycetes were conducted in permanent plots in beech forests, mostly in luzulo pilosae-fagetum, in the nature reserves wiączyń, gałków, and parowy janinowskie, as well as in the paprotnia forest. the reports on slime moulds from that area indicate a great role of dead wood, especially of beech trees, for slime moulds occurrence (seta, drozdowicz 2004; ślusarczyk revised data on the occurrence of myxomycetes in central poland 227 table 2 myxomycetes recorded in central poland name of species substrate source of information amaurochaete atra (alb. & schwein.) rostaf. ps 1; 2 a. tubulina (alb. & schwein.) t. macbr. fallen leaves of bp; stump of ps 1; 2 arcyria affinis rostaf. decayed trunk of fs 2 a. cinerea (bull.) pers. wood of deciduous trees 1; 2; 3; 4: b (30005, 30000, 30008); js (30004) a. denudata (l.) wettst. wood of deciduous trees 1; 2; 3; 4: b (300130); lubl (30028); lł (30014, 30016); msz (30022) a. ferruginea saut. wood of deciduous trees 1; 2; 4: lł (30032); s (30033) a. incarnata (pers. ex j.f. gmel.) pers. pa, ps 1; 2; 4: b (30038); lubl (30041) a. obvelata (oeder) onsberg wood of coniferous and deciduous trees 1; 2; 3; 4: arr (30048); b (30044, 30055, 30056); js (30062); lł (30050, 30052, 30053); sz (30054) a. pomiformis (leers) rostaf. pa, ps 1 badhamia affinis rostaf. bark of qu 2 b. capsulifera (bull.) berk. stump of ab 2 b. utricularis (bull.) berk. wood of deciduous trees; bark of ps; twigs of lar 2; 3 ceratiomyxa fruticulosa (o. f. müll.) t. macbr. wood of deciduous and coniferous trees 1; 2; 3; 4: b (30078, 30089); lubl (30073, 30076); lł (30072, 30074, 30077, 30092); s (30071) c. fruticulosa var. porioides (alb. & schwein.) g. lister pa, ps 1; 2 clastoderma debaryanum a. blytt ps 1 collaria arcyrionema (rostaf.) nann.-bremek. ex lado wood of deciduous trees 3; 4: lł (30302) comatricha nigra (pers. ex j. f. gmel.) j. schröt. fallen branches of deciduous trees; fs 2; 3 c. pulchella (c. bab.) rostaf. pa 1 craterium leucocephalum (pers. ex j. f. gmel) ditmar fallen leaves of qu 2 craterium minutum (leers) fr. fallen leaves of qu 4: lł (30748) cribraria argillacea (pers. ex j. f. gmel.) pers. wood of deciduous trees; cb, pa, ps 1; 2; 3 c. aurantiaca schrad. ps 4: gr (30126) c. cancellata (batsch) nann.bremek. wood of coniferous and deciduous trees 1; 4: b (30128, 3013, 30135, 30138); js (30127); lubl (30129) c. macrocarpa schrad. wood of coniferous trees 4: b (30749) c. rufa (roth) rostaf. ps 2; 4: lł (30143) c. tenella schrad. stump of ps 2 c. vulgaris schrad. pa 1; 2 diachea leucopodia (bull.) rostaf. herbaceous plants; fallen leaves of fs 2; 3; 4: js (30156, 30165, 30168) diderma radiatum (l.) morgan mosses 4: b (30169) d. spumarioides (fr.) fr. oa 4: s (30170) d. umbilicatum pers. mosses and fallen leaves 2 didymium iridis (ditmar) fr. bark of fs 1 d. leptotrichum (racib.) massee wood of deciduous trees 4: b (30171) d. melanospermum (pers.) t. macbr. mosses; needles of ps; wood of deciduous trees; pa 1; 2; 3; 4: lubl (30172) 4: lł (30174); sz (30175) 228 m. ławrynowicz et al. d. minus (lister) morgan stems of herbaceous plants 2 d. squamulosum (alb. & schwein.) fr. fallen leaves 2 enerthenema papillatum (pers.) rostaf. wood of deciduous trees 2 fuligo leviderma h.neubert, nowotny & k. baumann bp, fs 3 f. septica (l.) f.h. wigg. wood of deciduous and coniferous trees; qu, ps 2; 3; 4: arr (30200, 30212); ba (30210) b (30208, 30209, 30238); lł (30232 30232, 30235, 30236); msz (30196); mur (30222); s (30225); sz (30233) f. septica var. candida (pers.) r.e. fr. wood of deciduous and coniferous trees; ps 2; 4: arr (30259); b (30258, 30253); lubl (30254); lł (30256, 30257, 30260,); pap (30250); w (30252) hemitrichia clavata (pers.) rostaf. bp, fs 3; 4: lubl (30270, 30271); lł (30268) h. serpula (scop.) rostaf. ex lister ab 4: s (30710) lamproderma columbinum (pers.) rostaf. fs 1; 3 leocarpus fragilis (dicks.) rostaf. leaves of qu; stems of vm; fern leaves; bark of fs; bp fallen branches of qu, ps 1; 2; 3; 4: arr (30275); b (30274, 30277, 30285, 30295); lubl (30281); lł (30282, 30288, 30298) lindbladia tubulina fr. wood of deciduous trees 2 lycogala conicum pers. wood of coniferous trees; qu 1; 4: b (30306) l. epidendrum (l.) fr. wood of deciduous and coniferous trees; fs, bp, ps 1; 2; 3; 4: ba (30376); b (30359, 30395); j (30371); js (30396); l (30386); lubl (30341); lł (30352, 30353, 30354, 30382, 30387, 30388, 30397); msz (30356); mur (30343); s (30364, 30379, 30399); sz (30366) l. exiguum morgan ps 1 l. flavofuscum (ehrenb.) rostaf. wood of deciduous and coniferous trees 4: lł (30430) metarichia floriformis (schwein.) nann.-bremek. lar 4: lł (30435) m. vesparia (batsch) nann.bremek. ex g.w. martin & alexop. bp, ps; wood of deciduous trees 1; 2; 3; 4: b (30686, 30692); lubl (30691); lł (30690) mucilago crustacea f.h. wigg. grasses; fallen leaves of qu 1; 2 perichaena corticalis (batsch) rostaf. al 2 physarum album (bull.) chevall. fallen leaves of cb; qu; wood of deciduous and coniferous trees 1; 2; 3; 4: b (30470); lł (30455, 30469) ph. bivalve pers. fern leaves and fs 2; 4: js (30476); w (30475) ph. cinereum (batsch) pers. fs, ps 2; 3 ph. citrinum schumach. wood of deciduous and coniferous trees 4: b (30481, 30482, 30483) ph. compressum alb. & schwein. pt 1 ph. globuliferum (bull.) pers. lar, pa 1; 2 ph. gyrosum rostaf. pa, ps 1 ph. leucophaeum fr. wood of deciduous trees; fs 3; 4: b (30487) ph. notabile t. macbr. fs 3 ph. psittacinum ditmar stump of qu 2 table 2 – cont. revised data on the occurrence of myxomycetes in central poland 229 2010). several authors find beech wood as an excellent substrate for slime moulds (e.g., miśkiewicz 2001; stojanowska, panek 2004). the abundance of wood in different stages of decay supports effectively development of a variety of slime moulds species (stojanowska 1979; drozdowicz 1992). the results presented in the above mentioned papers as well as the studies of numerous collections from other areas of central poland confirm that dead wood is the most common substrate colonized by slime moulds. even in the area of łódź city and its surroundings decaying wood of different trees and shrubs was the basic substrate for them. in the tilio-carpinetum and potentillo albae-quercetum, the main substrate for slime moulds was also wood (trunks, logs, stumps, twigs) and fallen leaves of quercus and carpinus. it corresponds with reports by other authors, e.g., stojanowska & panek (2004) and salamaga & drozdowicz (2010). the variety of microhabitats offering different moisture, temperature and light conditions, decide of the slime ph. robustum (lister) nann. bremek. twigs of ca 2 ph. virescens ditmar qu, fallen leaves of fs; needles of ps 2; 3; 4: b (30496, 30503); js (30501) ph. viride (bull.) pers. wood of deciduous trees 2 ph. viride var. aurantium (bull.) lister wood of deciduous trees; stump of qu 2; 4: b (30509); js (30511) reticularia lycoperdon bull. wood of deciduous and coniferous trees; trunk of fs; qu, ps 1; 2; 3; 4: lł (30515) stemonitis axifera (bull.) t. macbr. wood of deciduous and coniferous trees; fs, bp, qu, ps 1; 2; 3; 4: b (30544); lł (30531, 30534, 30536, 30542); sz (30535) s. flavogenita e. jahn qu 2 s. fusca roth wood of coniferous trees; litter; al 1; 2; 3; 4: ba (30575); b (30561); lubl (30571) s. pallida wingate fs 3 stemonitopsis typhina (f.h. wigg.) nann.-bremek. wood of deciduous and coniferous trees; bark of qu; bp, ps 1; 2; 3; 4: b (30592); js (30712) trichia affinis de bary ps 1 t. contorta (ditmar) rostaf. wood of deciduous trees; litter 1; 4: msz (30597) t. decipiens (pers.) t. macbr. wood of deciduous trees; fs, bp 2; 3 t. favoginea (batsch) pers. wood of deciduous trees; fs 1; 3; 4: b (30608) t. persimilis p. karst. wood of deciduous trees; fs 2; 4: lubl (30272) t. scabra rostaf. bp, fs 2; 3; 4: b (30614) t. varia (pers. ex j.f. gmel.) pers. wood of deciduous trees; fs 1; 2; 3; 4: msz (30626) tubifera ferruginosa (batsch) j.f. gmel. wood of deciduous and coniferous trees; fs; litter; mosses 1; 2; 3; 4: b (30636, 30643); lł (30640, 30642, 30654); s (30646, 30649) explanations. source of information: 1 – orzechowski (1966); 2 – kalinowska-kucharska (1975); 3 – ślusarczyk (2010); 4 – herbarium material. wood of trees: ab – abies alba, al –alnus glutinosa, bp – betula pendula, ca – corylus avellana, cb – carpinus betulus, fs – fagus sylvatica, lar – larix decidua, oa – oxalis acetosella, qu – quercus sp., ps – pinus sylvestris, pt – populus tremula, tc – tilia cordata, vm – vaccinium myrtillus. table 2 – cont. 230 m. ławrynowicz et al. moulds species diversity. however, it is impossible to point the direct relationship of a particular plant association with myxomycete species composition. these observations correspond with those by other authors (e.g., stojanowska, panek 2002; panek, romański 2010). the wood of coniferous trees, especially of pinus sylvestris needs a special attention as a substrate for slime moulds. as the areas of occurrence of fagus sylvatica, abies alba and picea abies in central poland are among its most valuable natural sites, and they were most thouroughly examined in terms of myxomycetes diversity, the knowledge of slime moulds in central poland is mainly restricted to nature reserves. there are still a lot of other places that might be rich in myxomycetes and provide interesting information about this group of organisms, like various areas of other types of forests as well as sites of anthropogenic origin. it is difficult to make any generalizations based on the data summarized in the presented checklist, but a proper programme of research focused on slime moulds collections data could bring interesting and realistic results. conclusions the presented survey shows that the area of central poland is an attractive and valuable object of studies on myxomycetes. the results of the observations and collections that has been made for the last period of more than 50 years are an important input to the knowledge of slime moulds biodiversity. the checklist provided in the paper may serve as a source of information about the myxomycetes occurrence, species diversity, distribution and ecological relationships. it is a good starting point for further thorough qualitative and quantitative studies on slime moulds at present and in the future. acknowledgements. the authors would like to thank dr. anna drozdowicz (jagiellonian university) for her assistance in taxonomical interpretation of the collected materials and dr. marcin kiedrzyński (university of łódź) for preparing the map. references drozdowicz a. 1992. slime moulds (myxomycetes) of the ojców national park. iii. beech and fir logs as microhabitats of slime moulds. zeszyty naukowe uniwersytetu jagiellońskiego, prace botaniczne 24: 161–170. drozdowicz a., ronikier a., stojanowska w., panek e. 2003. myxomycetes of poland. a checklist. 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(in:) m. ławrynowicz, m. ruszkiewicz-michalska, i. kałucka (eds). traditional use and protection of fungi in poland: a contribution to the european cultural heritage. book of abstracts. wydawnictwo uniwersytetu łódzkiego, łódź: 130. 232 m. ławrynowicz et al. zweryfikowane dane o występowaniu śluzowców w polsce środkowej streszczenie kolekcja śluzowców z lat 1958-2009 z terenu polski środkowej przechowywana w herbarium uniwersytetu łódzkiego (lod f) była przedmiotem weryfikacji taksonomicznej z uwzględnieniem danych ekologicznych i chorologicznych. zbiory gromadzone były z różną intensywnością, najczęściej w czasie badań mykologicznych prowadzonych na terenach leśnych, głównie w rezerwatach przyrody chroniących stanowiska jodły, buka i świerka na granicy występowania tych drzew w polsce. na podstawie analizy 750 eksykatów zidentyfikowano 81 taksonów śluzowców. materiał pochodził z 34 stanowisk reprezentujących szerokie zróżnicowanie siedliska od rezerwatów przyrody do skrajnie zmienionych terenów w warunkach miejskich i podmiejskich łodzi. podsumowanie wyników zawiera krytyczna lista śluzowców polski środkowej. obecnie, w związku z podejmowaniem szczegółowych badań nad śluzowcami tego terenu, stanowić może punkt wyjścia do badań nad bioróżnorodnością, ekologią i rozmieszczeniem śluzowców polski środkowej oraz służyć do porównania z innymi obszarami obecnie i w przyszłości. 2014-01-01t23:50:39+0100 polish botanical society some endophytes of juncus trifidus from tatra mts. in poland andrzej chlebicki department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, a.chlebicki@botany.pl chlebicki a.: some endophytes of juncus trifidus from tatra mts. in poland. acta mycol. 44 (1): 11–17, 2009. this is a first part of work devoted highland rush endophytes: penicillium expansum, cladosporium oxysporum, arthrinium state of apiospora montagnei and aureobasidium pullulans. the basidiomycete strain, possibly lagarobasidium detriticum was also isolated. key words: highland rush, basidiomycete endophyte, arthrinium, penicillium, cladosporium, aureobasidium introduction asymptomal fungal infections within tissues of healthy plants is a basic character of endophytic fungi. these fungi can be dormant saprotrophs, latent pathogen as well as mutualistic symbionts (stone et al. 2000). sieber et al. (1991) noted that the latent endophyte phase is inversely related to the virulence of pathogen. fungal endophytes belong to different taxonomical groups of fungi such as ascomycetes and their anamorphs, mitosporic fungi and very rare basidiomycetous fungi. there are some tentatively using groups of endophytes as xylariaceous endophytes, dse (dark septate endophytes), aquatic mitosporic fungi, clavicipitaceous endophytes and others. of them fungi from the order clavicipitales are well known and evolutionary advanced endophytes (bacon, hill 1996). according to clay (1989) and white (1987) clavicipitaceous endophytes are widely distributed in the grass family poaceae and sporadically distributed in cyperaceae and juncaceae. other non-clavicipitaceous endophytes were noted in grasses (see larran et al. 2002) and rarely in rushes. according to powell (1975) rushes and sedges are non-mycotrophic, avoid of mycorrhiza. some endpohytes in rushes leaves of annual j. bufonius, perennial j. effusus, j. patens and j. bolanderi were reported by cabral et al. (1993). perennial rushes were inhabited by limited diversity of fungal species, mostly anamorphs and acta mycologica vol. 44 (1): 11–17 2009 12 a. chlebicki teleomorphs of dothidealen fungi, but only one or two species were recorded in high frequency. whereas annual rush was inhabited by distinctly diverse assemblage of mitosporic fungi including aquatic fungi with low isolation frequencies. according to cabral et al. (1993) rush endophytes were restricted to single epidermal host cell stagonospora innumerosa (desm.) sacc., substomatal cavity drechslera sp., cladosporium cladosporoides, alternaria alternata, and intercellular area of mesophyll tissue phaeosphaeria juncicola and alternaria alternata. men�ndez et al. (1997) not-men�ndez et al. (1997) not-(1997) noted platysporoides aff. togwotiensis (wehm.) shoemaker & c. e. babc. as endophyte in substomatal chamber of juncus imbricatus var. chamissonis. there are some articles devoted microfungi inhabiting different rushes. volkmann-kohlmeyer and kohlmeyer (1994) and kohlmeyer and volkmann-kohlmeyer (2000) described some marine fungi from saltmarsh juncus roemerianus. adamska (2005) noted six microfungi on various juncus species in lowland of poland. 30 taxa of microfungi (tab. i) were noted on highland rush by scheuer (1988, 1996, 1999); chlebicki (1990, 2002); scheuer & chlebicki (1997); suková (2004) and suková & chlebicki (2004) in the alps, carpathians, sudetes and ural mts. table 1 list of fungal species noted on plant organs of j. trifidus arthrinium cuspidatum lophodermium juncinum arthrinium state of apiospora montagnei mycosphaerella perexigua ascochyta junci naeviella paradoxa brikookea sepalorum niptera eriophori botrytis cinerea periconia atra brunnipila calycioides phaeosphaeria juncicola cladosporium herbarum phaeosphaeria vagans coronellaria caricinella phialocephala sp. cistella fugiens pseudoseptoria sp. dinemasporium strigosum pycnothyrium junci diplonaevia emergens septoria spp. epicoccum nigrum septoria chanousiana hysteronaevia minutissima stagonospora sp. hysteropezizella diminuens stagonospora junciseda lachnum dinminutum unguicularia sp. lachnum roseum only some endophytes were noted on specimens of juncus spp.: pleospora togwotiensis, stagonospora sp. on juncus imbricatus – (menéndez et al. 1997); stagonospora innumerosa, alternaria alternata, phaeosphaeria juncicola, drechslera sp., cladosporium cladosporioides, coniothyrium sp., exophiala salmonis, gyoerffyella craginiformis, spermospora –like sp., varicosporium elodeae, helicodendron spp., gliomastix sp., lambdosporium, phoma-like sp., titaea sp., acremonium (neotyphodium) sp., on j. bufonius, j. effuses, j. patens, j. bolanderi – (cabral et al. 1993). all these species are ascomycete fungi and its anamorphs, or mitosporic fungi. host plant. juncus trifidus l. (highland rush) belongs to the section steirochloa which is a sister group of luzula (drábkova et al. 2003). j. trifidus subsp. trifidus is the amphi-atlantic perennial plants of boreal-arctic-alpine distribution. the species is divided into three subspecies. of them j. trifidus subsp. carolinianus occurs in north america, j. trifidus subsp. monanthos is restricted to calcareous area of alps and appenines. the typical taxon j. trifidus subsp. trifidus occurs in silicate endophytes of juncus trifidus 13 mountains of europe, greenland and northern asia. the geographical distribution of the plant indicates its european origin (böcher 1972). plants from north america resembling those from greenland and scandinavia and differ in many characters from strains from pyrenees and tatra mts. (böcher 1972). during the pleistocene it spread in the arctic, reaching central siberia in the east (kulczyński 1924). in poland it occurs in some localities such as the karkonosze mts. (the sudetes), mt. babia góra range and tatra mts. (the carpathians). the locality in mt. śnieżnik in the central sudetes, reported by ciaciura (1988), is situated on the czech side of this mountain. highland rush is perennial caespitose rush forming dense tufts. aerial stems are stiff and erected, leaves alternate, simple with acuminate apices. inflorescence cymose or head-like. number of chromosomes of polish populations 2n=30 (skalińska et al.1957). it is pioneering alpine species with effective strategy of colonization of disturbed sites (marchand, roach 1980). methods material was collected on ne slope of mt. małołączniak in the tatra mts. (49˚ 55’n, 19˚ 14’e). 19 specimens of juncus trifidus were screened for the presence of fungal endophytes. small part of tuft including culms with rots were removed from the soil into sterile unclosed zip-lock plastic bags and putted in the refrigerator. after 3 days they were moving from the bags and processed according to isolation procedures (bacon 1990). three green culms of each sample were selected, central parts of each culm were cut into two segment ca 3 cm long. six segments from each sample were used (totally 114 segments). all segments were dipped for 1 min in 96% ethanol, then surface sterilized for 3 min in 3% chlorox (naocl), rinsed in ethanol for 1 min and transferred to 90 mm petri plates with ferency medium and then were incubated in room temperature in daylight. obtained isolates of different fungi were then removed on pda and maa media and incubated in room temperature. taxa of fungi were identified on the basis of cultural characteristics and the morphology of hyphae and conidia. results totally 114 segments were used. 11 strains of different fungi were isolated. of them five taxa are presented in this article. arthrinium state of apiospora montagnei sacc., nuovo giorn. bot. ital. 7: 306, 1875. colonies superficial, widely effused, produced mycelial mats, mycelium colourless to pale olivaceous brown, composed of network of septate hyphae anastomosing and branched, smooth ca 3 μm diam, conidiophores colourless, smooth 1.5-2 μm diam., conidia brown 5-9 × 4-5 μm with a hyaline band at the junction of the two 14 a. chlebicki sides. the fungus started growth after 5 months from two segments and overgrow every mycelia in the dishes. comments. the anamorph of this species was earlier noted by the author on leaves and stems of j. trifidus from ural mts. the colonies of this ural collection growing on stems, where circular, black with conidia 4.5-5(6) × 3.6-5 μm. it is cosmopolitan species, very common on bamboos and other dead plants (ellis 1971), with long hosts list (domsch, gams and anderson 1980). the fungi from the genus arthrinium were isolated from leaves of j. imbricatus var. chamissonis (menendez et al. 1995), bark of eucalyptus globulus (bettucci et al. 1999) and accidentally noted in tomato leaves (larran et al. 2001) and lichen talli of cladonia, stereocaulon (petrini et al. 1990) and xanthoparmelia taratica (as parmelia taratica girlanda et al. 1997). aureobasidium pullulans (de bary) g. arnaud, annals d’école national d’agric. de montpellier, n.s. 16: 39, 1918, var. pullulans. figs 1a1, a2 anamorph of discosphaerina höhn. after 10 days at the ends of rush segments appeared two pale colonies (fig. 1a1) which quickly joined and darken after 30 days (fig. 1a2). such black mycelium has been removed to the new plate with pda. colony black, convex, slightly pulvinate, surface covered by dense hyphae with conidia, margin entire to undulate, 21 mm diam. after 7 days. hyphae hyaline to pale brown, 4-5 μm diam., septate and branched, conidiogenous cells intercalary, conidia very variable in shape, (4)710(19) × (2)3-4 μm. comments. this black yeast-like fungus (meristematic fungus) is ubiquitous saprophyte of phyllosphere and other aerial plant parts (domsch, gams and anderson 1980). it is frequently noted as endophyte of different plants, among them in bark and xylem of eucalyptus (bettucci et al. 1999), leaf fern pteridium aquilinum (fisher 1996), twigs, leaves and buds of acer pseudoplatanus and other trees (pugh, buckley 1971). a. pullulans can be antagonistic against a number of phytopathogenic fungi (andrews et al. 1983). schena et al. (2002) noted high protection levels of a. pullulans against penicillium digitatum, botrytis cinerea and monilia laxa. a. pullulans is also noted on painted wood, stone and glass (yurlova et al. 1999). cladosporium herbarum (pers.) link, ges. naturf. freunde berlin mag. neuesten entdeck. gesammten naturk. 7: 37, 1816. anamorph of davidiella tassiana (de not.) crous & u. braun, mycol. progr. 2: 8, 2003. conidiophores straith to flexuose, distinctly nodose (were present on three stem segments in a single plate), conidia cylindrical, limoniform, pale olivaceous, nearly smooth or very slightly verrucose 19-16 × 5-6 (7) μm. comments. conidia of the species complex of c. herbarum have minutely verruculose to echinulate or spiny conidia (schubert et al. 2007). conidia of similar species c. oxysporum are generally smooth. it is known that species of cladosporium may inhabit substomatal chambers. shorter superficial sterilization times may lead to higher percentage of cladosporium. c. oxysporum was noted in bark and xylem of eucalyptus grandis (bettucci et al. 1999). cladosporium cladosporoides was noted as endophyte in branches of fraxinus excelsior and pinus sylvestris (kowalski, kehr 1996) and juncus spp. (cabral et al. 1993). c. herbarum was isolated as endophyte endophytes of juncus trifidus 15 in many different plants. suková (2004) noted this fungus on stems of j. trifidus in czech republik. penicillium expansum link, obs. mycol. 1: 16, 1809. figs 1b1, 1b2 anamorph of eupenicillium f. ludw. at the beginning colonies white (fig. 1b1), after becoming grey to olivaceousgrey on ferency medium (fig. 1b2). conidia pale yellow, smooth walled ellipsoidal 3-4 × 2,4-3 μm. comments. common fungus on fruits, meat, paper, various rotting substrates and soil (domsch et al. 1980), known as sorbus endophyte (samson, frisvad 2004). also batista et al. (2003) and vega & posada (2006) informed that fungi from the genera penicillium and aspergillus where isolated as endophytes. p. expansum strongly suppressed growth of black yeast strain growing on nearest segment. basidiomycete endophyte, possibly lagarobasidium detriticum (bourdot & galzin) jülich, persoonia 10(3): 334, 1979. syn.: hypochnicium detriticum (bourd. & galzin) j. erikss. & ryvarden, corticiaceae of north europe (oslo) 4: 701, 1976. the white mycelium was growing on five segments of culm. mycelium transferring on maa medium has 54 mm diam. after five days and 80 mm after seven days. mycelium white, velvety, pale yellow underside. at the beginning aerial mycelium cover all dish. hyphae 3-5 μm diam., with clamps, spores smooth, ellipsoid 6-6.5 × 5-5.5 μm diam. with a small apiculus and big oil drop. crystals not seen. cystidia not present, however similar structures were seen. comments. size of spores resemble l. detriticum. eriksson and ryvarden (1976) noted l. detriticum on leaves of juncus, carex, eqisetum, ferns, on rotten woods of juniperus, alnus, betula, populus and sorbus. basidiomycete endophytes are not often isolated from plants. some species from the genus peniophora were isolated from xylem and bark of eucalyptus globulus and they probably initiated the decomposition process (bettucci et al. 1999). discussion totally 11 strains of different fungi were isolated. of them a black yeast, basidiomycete fungus, dse and non-sporulating colonies will be subject of the next article. in this article are presented first result of presence of endophytes in the highland rush stems. four isolated species of fungi have worldwide general distribution and were earlier noted as endophytes of various plants. the precise determination of the most interesting basidiomycete strain need sequencing of its region (nilson 2003). also comparison of frequency of various ecological groups of endophytes needs further investigation. so far any endophytes were isolated from stems of highland rush. acknowledgements. i especially thank to anonymous reviewer for important comments. this work was supported by the ministry of science and information society technologies, poland (project no. 2 p04f 066 28 and n n304 328336). 16 a. chlebicki references adamska i. 2005. fungal species new in poland on carex and juncus. acta mycol. 40: 19–24. andrews j. h., berbee f. m., nordheim e. f. 1983. microbial antagonism to the imperfect stage of the apple scab pathogen, venturia inaequalis. phytopathology 73: 228–234. bacon c.w. 1990. isolation, cultures, and maintenance of endophytic fungi of grasses. 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(in:) c. w. bacon, j. f. white (eds). microbial endophytes. marcel dekker, new york, basel: 3–29. suková m. 2004. fungi on juncus trifidus in the czech republic. i. czech mycol. 56: 63-84. suková m., chlebicki a. 2004. fungi on juncus trifidus in the czech republic (ii) with taxonomical notes to some species. czech mycol. 56: 203–221. vega f. e., posada f. 2006. penicillium species endophytic in coffee plants and ochratoxin a production. my-mycologia 98 (1): 31–42. volkmann-kohlmeyer b., kohlmeyer j. 1994. a new aniptodera from saltmarsh juncus. botanica marina 37: 109–113. white j.f. jr. 1987. widespread distribution of endophytes in the poaceae. plant dis. 71: 340–342. yurlova n. a., de hoog g. s., gerrits van den ende a. h. g. 1999. taxonomy of aureobasidium and allied genera. studies in mycology 43: 63–69. kilka endofitycznych grzybów wyizolowanych z pędów situ skuciny (juncus trifidus) z polskiej części tatr streszczenie do badań wybrano populację situ skuciny rosnącą w masywie czerwonych wierchów w polskiej części tatr. pobrano materiały 19 prób pędów, wszczepiono 114 fragmentów pędów. uzyskano 11 szczepów, z czego 5 opisano w niniejszym artykule: arthrinium, anamorfa apiospora montagnei, cladosporium herbarum, pennicillium expansum, aureobasidium pullulans i szczep należący do grzybów podstawkowych, prawdopodobnie lagarobasidium detriticum. fig. 1. aureobasidium pullulans: a 1 – after 10 days; a 2 – after 30 days; penicillium expansum: b1 – after 10 days; b2 – after 30 days. both strains were cultivated on ferency medium. 2014-01-01t11:48:35+0100 polish botanical society new and interesting records of freshwater verrucaria in central poland beata krzewicka1 and mariusz hachułka2 1laboratory of lichenology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, ibbkrzew@ib-pan.krakow.pl 2laboratory of lichenology, department of mycology, university of łódź banacha 12/16, pl-90-237 łódź, m.hachulka@poczta.fm krzewicka b., hachułka m.: new and interesting records of freshwater verrucaria in central poland. acta mycol. 43 (1): 91–98, 2008. verrucaria madida is reported as new to poland. three other associated species, v. aquatilis, v. hydrela and v. rheitrophila, are compared. the known distribution in poland and the ecology of these freshwater species are presented. key words: lichenized fungi, pyrenocarpous lichen, verrucaria madida introduction during studies on the lichen biota of the wzniesienia łódzkie landscape park (central poland) carried out by the second author, a very interesting body of material was found on submerged rocks in a watercourse. as the examination showed, individuals observed on the rocks belong to verrucaria madida orange species. the taxon is rare in europe and was not reported from poland. the species was associated with v. aquatilis, v. hydrela, and v. rheitrophila in the study area. except v. aquatilis and v. rheitrophila, the species are recorded from central poland for the first time (nowak 1967; hachułka 2007). study area the wzniesienia łódzkie landscape park (total area of 13 767 ha) is located in central poland between łódź, stryków and brzeziny (fig. 1) within the fringe zone of the wzniesienia łódzkie elevation mesoregion (kondracki 2000). springs of upland streams: grzmiąca, kamienna as well as moszczenica and its tributary, młynówka, lie in the park. the grzmiąca and kamienna are tributaries of the mrożyca, whose springs are outside the park borders. the streams form the catchment area of the bzura river and flow along incised valleys (from 2‰ to 4.5‰) in the spring section acta mycologica vol. 43 (1): 91–98 2008 new and interesting records 97 references bielczyk u. 2003. the lichens and allied fungi of the polish western carpathians. 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(ed.). 1998. park krajobrazowy wzniesień łódzkich. monografia. wyd. eko-wynik, łódź, 182 pp. moniewski p. 2004. źródła okolic łodzi. acta geogr. lodz. 87: 1–140. nowak j. 1965. porosty beskidu małego. fragm. flor. geobot. 11 (3): 209–242. nowak j. 1967. porosty wyżyny wieluńskiej. acta mycol. 3: 209–242. nowak j. 1998. porosty beskidów wyspowego i żywieckiego, pasma jałowca i masywu babiej góry. monogr. bot. 83: 1–131. orange a. 2004. a remarkable new freshwater verrucaria from europe. lichenologist 36 (6): 349–354. raport o stanie środowiska w województwie łódzkim w 2004 roku. 2005. wojewódzki inspektorat ochrony środowiska w łodzi. oficyna wydawnicza, łódź: 116–143. szymczyk r. 2007. rzadkie i interesujące gatunki porostów i grzybów naporostowych na wysoczyźnie elbląskiej (północna polska). fragm. flor. geobot. pol. 14 (1): 167–173. 98 b. krzewicka and m. hachułka nowe dane o wodnych gatunkach verrucaria z polski środkowej s t r e s z c z e n i e w pracy przedstawiono cztery gatunki porostów z rodzaju verrucaria: v. aquatilis, v. hydrela, v. madida i v. rheitrophila, zebrane w strumieniach zlokalizowanych w parku krajobrazowym wzniesień łódzkich. materiał zebrany w roku 2006 przez m. hachułkę został zrewidowany lub oznaczony przez pierwszego autora. verrucaria madida jest gatunkiem nowym dla bioty polski. porost ten ma plechę ciemno-zieloną do zielonawoczarnej, inwolukrelum stożkowate lub po bokach rozpościerające się i wówczas lekko powyginane (faliste), worki z (3-)4(-5) zarodnikami; zarodniki jednokomórkowe elipsoidalne 12(-17) x 7 μm. według orange (2004) v. madida znany jest z kilku rozproszonych stanowisk w europie północno-zachodniej, z wielkiej brytanii (walia), francji oraz z norwegii. w parku krajobrazowym gatunek ten, wraz z towarzyszącymi mu v. aquatilis, v. hydrela i v. rheitrophila odnotowano w odcinku źródłowym w młynówce na pierwszym stanowisku, na kwaśnych kamieniach zanurzonych w wodzie. v. hydrela występuje na wszystkich badanych stanowiskach i jest podawana po raz pierwszy z polski środkowej. v. rheitrophila odnotowano w młynówce na pierwszym i trzecim stanowisku oraz w kamiennej. ostatni gatunek był podawany z polski środkowej jedynie przez nowaka (1967) z wyżyny wieluńskiej (łącznie z v. aquatilis) i hachułkę (2007) ze wzniesień łódzkich. 2014-01-01t11:47:28+0100 polish botanical society fungi isolated from the rhizosphere of spring cruciferous plants barbara majchrzak1, adam okorski2 and bogusław chodorowski1 1department of phytopathology and entomology, university of warmia and mazury prawocheńskiego 17, pl-10-722 olsztyn 2department of diagnostics and plant pathophysiology university of warmia and mazury, plac łódzki 6, pl-10-727 olsztyn, adam.okorski@uwm.edu.pl majchrzak b., okorski a., chodorowski b.: fungi isolated from the rhizosphere of spring cruciferous plants. acta mycol. 43 (2): 181–191, 2008. fungal communities isolated from the rhizosphere of spring cruciferous plants were analysed in the study. it was found that the rhizosphere of crucifers was colonized primarily by fungi of the order mucorales and of the genus fusarium. members of the genus fusarium dominated in the rhizoplane. the roots of cruciferous plants secrete glucosinolates – secondary metabolites known for their antifungal properties, thus affecting the communities of soil-dwelling fungi. key words: rhizosphere, cruciferous plants, fungi, fusarium introduction the soil provides habitat for both phytopathogenic and saprotrophic microorganisms, including bacteria, actinomycetes and fungi (patkowska 1998). the rhizosphere, i.e. the zone that surrounds the roots of plants, plays a particularly important role due to its specific biological properties. it is teeming with a wide variety of microbes (morgan et al. 2005), which can be divided into plant growth-promoting rhizobacteria (pgpr), deleterious rhizosphere microorganisms (drmo) and neutral microorganisms – having no impact on plant growth (sturz, christie 2003). under natural conditions, in undisturbed soil, the groups of beneficial and harmful microorganisms remain in the state of dynamic equilibrium. pgpr contribute to yield increment, usually resulting from higher nutrient availability and suppression of the growth and activity of deleterious microorganisms. drmo compete with pgpr for food, thus negatively affecting crop development (kurek, kobus 1990). due to their antagonist potential, soil microorganisms are able to colonize suitable niches. the antagonistic mechanisms include antibiosis, competition and mycoparasitism (hyperparasitism) (sturz, christie 2003). many pathogens develop in the after-harvest residues of forecrops, so the type of forecrop may have a significant influence on the acta mycologica vol. 43 (2): 181–191 2008 182 b. majchrzak et al. yield of successive crops (bojarczuk, bojarczuk 1988). crucifers and legumes play a positive role in crop rotation because they improve the chemical, physical and biological properties of the soil (majchrzak et al. 2002). members of the fa mily brassicaceae are among the best forecrops, because they leave in the soil large amounts of after-harvest residues rich in glucosinolates and other secondary metabolites (oleszek 1997). moreover, they exert a positive effect on the health of successive crops (majchrzak et al. 2004; majchrzak et al. 2005). the objective of this study was to determine the species composition of fungi isolated from the rhizosphere of selected cruciferous plants. materials and methods the study was conducted during the years 1999-2001 at the production-experimental station in bałcyny near ostróda (ne poland), on the experimental plots of the department of plant production, university of warmia and mazury in olsztyn. the experiment was established on gray-brown podsolic soil developed from light silty clay, of quality class iii a, of good wheat complex (1999 and 2000) or very good rye complex (2001). crucifers were grown after spring wheat in 1999 and 2000, and after winter rapeseed in 2001. mineral fertilizers (npk) were applied at the following rates: 60-100 : 40-60 : 60-100 kg•ha-1, as recommended by the institute of soil science and plant cultivation. the field trial was performed in a randomized block design, in three replications. the experimental factors were as follows: factor i – cruciferous plants spring oilseed rape (• brassica napus f. annua) – cv. margo, white mustard (• sinapis alba) – cv. heter, chinese mustard• (brassica juncea var. sareptana) – cv. małopolska, radish (• raphanus sativus var. oleifera) – cv. pegletta, false flax (• camelina sativa) – cv. borowska, spanish colewort (• crambe abyssinica)cv. borowski factor ii – years of the study no fungicides were applied. fungi were isolated from the rhizosphere, rhizoplane and roots of crops as described by mańka (1974). the quantitative and qualitative composition of fungal communities was determined at full blooming (bbch 65-69). results a total of 2 929 fungal colonies belonging to 99 species and non-spore forming fungi were isolated from the rhizosphere of spring cruciferous plants over the three-year experimental period (tab.1). members of the order mucorales dominated among them (48.96% of all colonies). representatives of the genus rhizopus were isolated most frequently (15.94%). fungi of the genus penicillium were also abundant (12.43%). antagonistic species, including the order mucorales and the genera gliocladium, penicillium and trichoderma, accounted for 66.58% of all isolates. the proportion of pathogens in the rhizosphere was 11.33%, and the predominant role was played by species of the genus fusarium (8.77%), primarily f. solani, f. oxysporum and f. equiseti. fungi isolated from the rhizosphere 183 the most diverse fungal community, composed of 637 colonies representing 44 species, was isolated from the rhizosphere of spring rape (tab. 1). mucorales (46.10%), fusarium (16.5%), penicillium (7.85%) and gliocladium (6.12%) dominated among them. the fewest fungi (339 isolates belonging to 36 species) were detected in the rhizosphere of false flax. the most abundant among them were: mucorales – 57.8%, acremonium – 9.44% and fusarium – 6.49%. fungi of the genus fusarium were isolated least frequently from the rhizosphere of spanish colewort (3.81%). the genus penicillium (270 isolates – 46.80%) dominated in the rhizosphere of this crop. the rhizospheres of white mustard, spanish colewort and radish were characterized by the lowest species diversity (35 fungal species each). a total of 568 fungal colonies belonging to 80 species and non-spore forming fungi were isolated from the rhizoplane of spring cruciferous plants during the experimental period (tab. 2). the most diverse fungal community, comprising 134 colonies representing 34 species, was isolated from the rhizosphere of spring rape. members of the genera fusarium (32.84%) and acremonium (21.64%) as well as of the order mucorales (5.69%) dominated among them. the fewest fungi were isolated from the rhizoplane of spanish colewort (73 isolates). this community, composed of only 24 species, was found to be the least diverse. the rhizoplane of this crop was mostly colonized by gliocladium spp. (21.90%), aspergillus fumigatus (15.10%) and penicillium spp. (15.10%). representatives of the genus fusarium constituted the least numerous group in the rhizoplane of spanish colewort (6 isolates – 8.22%). discussion research results show that plants of the genus brassicaceae grown as forecrops or ploughed in as green manure have a beneficial effect on the health of field crops (majtahedi et al. 1991). the roots of crucifers secrete glucosinolates, which affect the soil microflora and help to control the occurrence of phytopathogens (bones, rossiter 1996; kierkegaard, sarwar 1998). decomposition of the tissues of brassicaceae as well as the production of glucosinolates followed by their hydrolysis lead to the formation of isothiocyanates (itcs) – volatile substances considered to be biofumigants (sarwar et al. 1998). according to snapp et al. (2007) and charron and sam (1999), growing plants of the genus brassicaceae as forecrops and leaving their remainders in the field inhibits the growth of such soil pathogens as rhizoctonia solani and pythium ultimum. marwar and lodha (2002) demonstrated that plants of the family brassicaceae limited the occurrence of fusarium oxysporum f. sp cumini. in the present study fungi of the genus fusarium were not abundant in the rhizosphere of crucifers. their population was considerably greater in the rhizoplane. the soil environment of particular cruciferous plants was colonized by members of this genus to a different degree. fusarium colonies were isolated most frequently from the rhizosphere and rhizoplane of spring oilseed rape, and least frequently from the rhizosphere and rhizoplane of spanish colewort and false flax. ishimoto et al. (2000) reported that fungi of the genus fusarium isolated from the roots of crucifers showed high tolerance to glucosinolates, which may suggest that they acquired resistance to this group of substances through adaptation. in the current experiment the rhizosphere of the cruciferae was colonized by numerous representatives of the order mucorales, dominated by members of the 184 b. majchrzak et al. t a b le 1 fu ng i i so la te d fr om r hi zo sp he re s pr in g cr uc if er ou s pl an ts fu ng al s pe ci es p la nt s su m % o ils ee d ra pe w hi te m us ta rd c hi ne se m us ta rd o ils ee d ra pe fa ls e fla x sp an is h co le w or t 1* 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 a cr em on iu m b re ve ( su ka p. e t t hi ru m )w . g am s 33 2 35 1. 19 a cr em on iu m c ha rt ic ol a (l in da u) w . g am s 21 3 4 3 11 6 48 1. 64 a cr em on iu m c hr ys og en um ( su ka p. & t hi ru m )w . g am s 2 2 0. 07 a cr em on iu m c ur vu lu m w . g am s 16 6 6 28 0. 96 a cr em on iu m k ili en se g ru et z 15 8 23 0. 79 a cr em on iu m la rv ar um ( pe tc h) w . g am s 14 14 0. 48 a cr em on iu m m in ut is po ru m ( su ka p. e t t hi ru m )w . g am s 6 6 0. 20 a cr em on iu m o ch ra ce um ( o ni on s et b ar ro n) w . g am s 1 1 0. 03 a cr em on iu m p ot ro ni v ui ll 1 4 5 0. 17 a cr em on iu m p sa m m os po ru m w . g am s 4 2 2 11 19 0. 65 a cr em on iu m s cl er ot ig en um ( f. e t r . m or ea u ex v al en ta ) w . g am s 1 1 0. 03 a cr em on iu m s tr ic tu m w . g am s 3 3 0. 10 a cr em on iu m s p. 1 1 0. 03 a lte rn ar ia a lte rn at a (f ri es ) k ei ss le r 1 4 1 27 5 3 1 42 1. 43 a rt hr in iu m s ph ae ro sp er m um f uc ke l 1 3 4 0. 14 a sp er gi llu s re pe ns d e b ar y 1 1 0. 03 a ur eo ba si di um b ol le yi s pr ag ue 2 2 1 5 0. 17 a ur eo ba si di um p ul lu la ns d e b ar y 14 7 7 3 31 1. 06 b ot ry od ip lo di a sp . 1 1 0. 03 b ot ry tis c in er ea p er so on 3 1 4 8 0. 27 c ep ha lo sp or iu m a tr um d e b ar y 5 5 0. 17 c er co sp or a sp p. 5 5 0. 17 c la do sp or iu m c la do sp or oi de s (f re s. ) de v ri es 18 10 6 2 8 10 15 5 74 2. 53 c la do sp or iu m h er ba ru m l in k ex f r . 2 1 11 2 16 0. 55 c ol le to tr ic hu m s pp . 5 5 0. 17 c on io th yr iu m s pp . 1 5 3 4 13 0. 44 c yl in dr oc ar po n de st ru ct an s (z in s. ) sc ho lt en 4 6 2 2 1 15 0. 51 e nd ot hi a sp p. 3 3 0. 10 e pi co cc um p ur pu ra sc en s e hr en b. e x sc hl ec ht 1 1 0. 03 e pi co cc um s pp . 1 1 0. 03 fu sa ri um a ve na ce um c or da e x fr . 1 3 2 1 9 16 0. 55 fu sa ri um c hl am yd os po ru m ( w ol le nw eb er e t r ei nk in g) 2 2 4 0. 14 fu sa ri um c ul m or um ( w .g . s m it h) s ac c. 1 1 2 0. 07 fungi isolated from the rhizosphere 185 fu ng al s pe ci es p la nt s su m % o ils ee d ra pe w hi te m us ta rd c hi ne se m us ta rd o ils ee d ra pe fa ls e fla x sp an is h co le w or t 1* 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 fu sa ri um d im er um ( pe nz ig ) 7 4 11 0. 38 fu sa ri um e qu is et i ( c or da ) sa cc . 5 10 4 3 1 12 5 2 1 3 46 1. 57 fu sa ri um n iv al e (f r. ) c es . 1 13 1 1 16 0. 55 fu sa ri um o xy sp or um (s ch le ch t.) 13 29 2 7 2 8 1 12 4 1 4 9 2 94 3. 21 fu sa ri um s am bu ci nu m f uc k. 3 3 0. 10 fu sa ri um s ol an i ( m ar t.) s ac c. 18 1 2 17 1 2 1 4 7 1 4 58 1. 98 fu sa ri um s ol an i v ar . c oe ru le um t hu m 2 2 0. 07 fu sa ri um ta ba ci nu m ( b ey m a) w . g am s 4 4 0. 14 fu sa ri um s p. 1 1 0. 03 g lio cl ad iu m c at en ul at um g ilm an e t a bb ot t 1 2 10 2 11 26 0. 89 g lio cl ad iu m fi m br ia tu m g ilm an e t a bb ot t 4 5 9 0. 31 g lio cl ad iu m p en ic ill oi de s c or da 11 22 12 7 2 1 4 59 2. 01 g lio cl ad iu m s al m on ic ol or r ai llo 1 1 0. 03 g lio m as tix c er ea lis ( k ar t.) d ic ki ns on 1 1 0. 03 g lio m as tix m ur or um ( c or da ) h ug he s 1 1 0. 03 h um ic ol a fu sc oat ra t ra ae n 5 2 3 11 1 6 28 0. 96 h um ic ol a br ev is g ilm an e t a bb ot t 5 4 5 14 0. 48 h um ic ol a ni gr es ce ns o m vi k 1 2 3 1 1 8 0. 27 k ic kx el la a la ba st ri na c oe m an s 1 1 0. 03 m on oc ili um m uc id um w . g am s 2 2 0. 07 m on od ic tis le vi s (w ilt sh .) h ug he s 2 2 2 6 0. 20 m or tie re lla a lli ac ea l in n 1 1 0. 03 m or tie re lla e lo ng at a l in ne m an n 2 2 0. 07 m or tie re lla g em m ife ra e lli s 1 1 0. 03 m or tie re lla g ra ci lis l in ne m an n 7 7 0. 24 m or tie re lla m ar bu rg en si s l in ne m an n 2 2 4 0. 14 m or tie re lla a lp in a pe yr on d 13 7 10 30 1. 02 m or tie re lla v in ac ea d ix on -s te w ar t 1 1 0. 03 m or tie re lla s pp . 30 4 1 1 96 13 2 4. 51 m uc or c ir ci ne llo id es v an t ie gh em 2 2 0. 07 m uc or h ie m al is w eh m er 86 36 7 72 14 2 25 3 11 11 5 11 15 5 30 18 67 5 23 .0 5 m uc or m ic ro sp or us n am ys lo w sk ii 6 6 0. 20 m uc or m uc ed o (l in ne ) b re fe ld 3 2 5 10 0. 34 m uc or p ir ifo rm is f is ch er 37 37 1. 26 m uc or p us ill us l in dt 23 5 28 0. 96 m uc or ra ce m os us f re se ni us 8 7 5 11 31 1. 06 pa ec ill om yc es v ar ia bi lis b ar ro n 1 1 0. 03 186 b. majchrzak et al. fu ng al s pe ci es p la nt s su m % o ils ee d ra pe w hi te m us ta rd c hi ne se m us ta rd o ils ee d ra pe fa ls e fla x sp an is h co le w or t 1* 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 pe ni ci lli um n ig ri ca ns ( b ai n. ) t ho m 2 1 3 0. 10 pe ni ci lli um s pp . 35 15 12 5 1 16 2 1 4 1 23 0 39 36 1 12 .3 3 pe ri co ni a fu ne re a (c es .) m as on e t e lli s 2 2 0. 07 p ho m a eu ry pe na s ac c. 9 6 15 0. 51 p ho m a fim et i ( b ru n) 9 2 11 0. 38 p ho m a gl om er at a (c or da ) w ol le nw eb er e t h oc ha pf el 2 2 0. 07 p ho m a he rb ar um w es te nd . 2 2 0. 07 r hi zo ct on ia s ol an i k üh n 8 8 0. 27 r hi zo ct on ia s pp . 1 1 2 0. 07 r hi zo pu s ni gr ic an s e hr en be rg 1 6 2 9 0. 31 r hi zo pu s sp p. 11 3 71 39 39 5 15 2 39 45 8 15 .6 4 sc le ro tin ia s cl er ot io ru m ( l ib .) d e b ar y 15 15 0. 51 sc op ul ar io ps is a cr em on iu m ( d el ac r. ) v ui ll. 2 1 3 0. 10 sc op ul ar io ps is b re vi ca ul is ( sa cc .) b ai n 13 13 0. 44 sc yt al id iu m li gn ic ol a pe sa nt e 4 3 7 0. 24 sp ic ar ia c ar no sa m ill er .g id de ns e t f os te r 1 1 0. 03 sp ic ar ia d iv ar ic at a (t ha n. ) g ilm an & a bb ot t 7 7 0. 24 sp ic ar ia g ri se ol a sa cc ar do 9 9 0. 31 sp ic ar ia s im pl ic is si m a o ud em an s 1 1 0. 03 sp ic ar ia v io la ce a a bb ot t 41 32 73 2. 49 sp or ot ri ch um c ar ni s b ro ok s et h an sf or d 1 1 2 0. 07 sp or ot ri ch um c hl or in um l in k 1 1 0. 03 sp or ot ri ch um o liv ac eu m f ri es 2 2 0. 07 to ru la h er ba ru m ( pe rs .) l in k ex f r. 5 5 0. 17 tr ic ho de rm a au re ov ir id e r if ai 10 5 2 17 0. 58 tr ic ho de rm a ha m at um ( b on .) b ai n 6 2 5 1 1 1 16 0. 55 tr ic ho de rm a ha rz ia nu m r if ai 1 10 8 19 0. 65 tr ic ho de rm a po ly sp or um ( l in k et p er s. ) r if ai 5 5 0. 17 ve rt ic ill um c el lu lo sa e d as ze w sk a 3 3 0. 10 n on -s po ru la ti ng fu ng i 1 1 4 4 5 1 5 5 6 32 1. 09 su m 98 29 9 24 0 10 6 20 3 10 7 29 33 9 19 0 87 18 9 12 6 57 88 19 4 67 36 8 14 2 29 29 10 0. 0 su m o f p la nt 63 7 41 6 55 8 40 2 33 9 57 7 *1 19 99 2 2 00 0 3 20 01 t a b le 1 fu ng i i so la te d fr om r hi zo sp he re s pr in g cr uc if er ou s pl an ts – c on t. fungi isolated from the rhizosphere 187 t a b le 2 fu ng i i so la te d fr om r hi zo pl an e of s pr in g cr uc if er ou s pl an ts fu ng al s pe ci es p la nt s su m % o ils ee d ra pe w hi te m us ta rd c hi ne se m us ta rd o ils ee d ra pe fa ls e fla x sp an is h co le w or t 1* 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 a cr em on iu m b re ve ( su ka p. & t hi ru m ) w . g am s 8 8 1. 41 a cr em on iu m c ha rt ic ol a (l in da u) w . g am s 2 1 2 2 1 8 1. 41 a cr em on iu m c hr ys og en um ( su ka p. e t t hi ru m ) w . g am s 1 1 2 0. 35 a cr em on iu m c ur vu lu m w . g am s 7 7 1. 23 a cr em on iu m in co lo ra tu m ( su ka p. e t t hi ru m ) w . g am s 1 1 0. 18 a cr em on iu m k ili en se g ru et z 1 1 3 5 0. 88 a cr em on iu m la rv ar um ( pe tc h) w . g am s 1 1 0. 18 a cr em on iu m m in ut is po ru m ( su ka p. e t t hi ru m ) w . g am s 1 1 2 0. 35 a cr em on iu m p ot ro ni v ui ll 2 1 2 5 0. 88 a cr em on iu m p sa m m os po ru m w . g am s 1 1 2 0. 35 a cr em on iu m s tr ic tu m w . g am s 1 1 2 0. 35 a cr em on iu m s pp . 3 1 1 1 3 9 1. 58 a cr os pe ir a m ir ab ili s b er k. e t b r. 2 2 0. 35 a lte rn ar ia a lte rn at a (f ri es ) k ei ss le r 1 1 1 9 1 2 2 17 2. 99 a sp er gi llu s fu m ig at us f re se ni us 1 11 12 2. 11 a sp er gi llu s fu ni cu lo su s (g . s m it h) 1 1 0. 18 a sp er gi llu s re pe ns d e b ar y 1 1 0. 18 a sc oc hy ta s pp . 1 2 3 0. 53 a ur eo ba si di um b ol le yi ( sp ra gu e) 1 1 1 1 1 3 1 9 1. 58 a ur eo ba si di um p ul lu la ns d e b ar y 1 6 1 3 1 1 1 2 2 1 19 3. 35 b ot ry ot ri ch um p ilu lif er um s ac ca rd o et m ar ch al 1 1 0. 18 b ot ry tis c in er ea p er so on 1 2 3 0. 53 c la do sp or iu m c la do sp or oi de s (f re s. ) de v ri es 1 2 1 1 4 4 1 1 6 1 22 3. 87 c la do sp or iu m h er ba ru m ( pe rs oo n) l in k 1 1 3 1 6 1. 06 c on io th yr iu m s pp . 1 1 2 0. 35 c yl in dr oc ar po n de st ru ct an s (z in s. ) sc ho lt en 2 1 1 1 1 1 7 1. 23 e nd ot hi a sp . 1 1 0. 18 e pi co cc um s p. 1 1 0. 18 fu sa ri um a ve na ce um ( c or da e x fr .) 1 1 1 1 4 0. 70 fu sa ri um c hl am yd os po ru m ( w ol le nw eb er e t r ei nk in g) 2 1 1 1 5 0. 88 fu sa ri um c ul m or um ( w .g . s m it h) s ac c. 1 1 1 1 1 5 0. 88 fu sa ri um e qu is et i ( c or da ) sa cc . 6 1 3 1 3 1 15 2. 64 188 b. majchrzak et al. fu ng al s pe ci es p la nt s su m % o ils ee d ra pe w hi te m us ta rd c hi ne se m us ta rd o ils ee d ra pe fa ls e fla x sp an is h co le w or t 1* 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 fu sa ri um fu sa ri oi de s (f ra g. c if .) 1 1 0. 18 fu sa ri um o xy sp or um ( sc hl ec ht .) 3 1 1 1 3 2 3 5 2 21 3. 70 fu sa ri um s ol an i ( m ar t.) s ac c. 1 15 1 4 3 4 2 2 2 3 2 1 40 7. 04 fu sa ri um s ol an i v ar . c oe ru le um t hu m 1 7 1 9 1. 58 fu sa ri um ta ba ci nu m ( b ey m a) w . g am s et a bb ot t 1 1 2 0. 35 g lio cl ad iu m c at en ul at um g ilm an e t a bb ot t 8 1 1 1 1 2 14 2. 46 g lio cl ad iu m fi m br ia tu m g ilm an & a bb ot t 1 13 14 2. 46 g lio cl ad iu m p en ic ill oi de s c or da 2 1 2 1 6 1. 06 g lio m as tix m ur or um ( c or da ) h ug he s 1 1 0. 18 h um ic ol a fu sc oat ra t ra ae n 1 4 2 2 3 1 2 1 16 2. 82 h um ic ol a br ev is g ilm an e t a bb ot t 1 1 2 0. 35 h um ic ol a ni gr es ce ns o m vi k 2 1 3 0. 53 k ic kx el la a la ba st ri na c oe m an s 1 1 0. 18 m ic ro do ch iu m n iv al e (f r. ) c es . 11 1 2 14 2. 46 m on oc ili um a rc tii co la ( w . g am s) 1 1 0. 18 m or tie re lla a lp in a pe yr on d 1 1 1 3 0. 53 m or tie re lla a rc ua ta w ol f 1 1 0. 18 m or tie re lla is ab el in a o ud em an s 1 1 0. 18 m or tie re lla v in ac ea d ix on -s te w ar t 1 1 0. 18 m uc or c ir ci ne llo id es v an t ie gh em 11 11 1. 94 m uc or h ie m al is w eh m er 3 2 7 30 2 2 1 3 1 1 52 9. 15 m uc or m uc ed o (l in ne ) b re fe ld 2 1 3 0. 53 m uc or p ir ifo rm is f is ch er 2 2 0. 35 m uc or ra ce m os us f re se ni us 1 2 1 4 0. 70 pa ec ili om yc es v ar io tii b ai ni er 1 1 0. 18 pa ec ilo m yc es n iv eu s st ol k et s am so n 2 2 0. 35 pe ni ci lli um n ig ri ca ns ( b ai n. ) t ho m 1 1 0. 18 pe ni ci lli um s pp . 2 8 7 2 2 7 4 1 4 1 8 2 48 8. 45 p ho m a ch ry sa nt em ic ol a h ol lo s 1 1 0. 18 p ho m a eu ry pe na s ac c. 2 2 1 5 0. 88 p ho m a gl om er at a (c or da ) w ol le nw eb er e t h oc ha pf el 2 2 0. 35 p ho m a he rb ar um w es te nd . 1 1 0. 18 t a b le 2 fu ng i i so la te d fr om r hi zo pl an e of s pr in g cr uc if er ou s pl an ts fungi isolated from the rhizosphere 189 fu ng al s pe ci es p la nt s su m % o ils ee d ra pe w hi te m us ta rd c hi ne se m us ta rd o ils ee d ra pe fa ls e fla x sp an is h co le w or t 1* 2 3 1 2 3 1 2 3 1 2 3 1 2 3 1 2 3 p yr en op ho ra s em en ip er da ( b ri tt le ba nk e t a da m ) 1 1 1 1 4 0. 70 r hi zo ct on ia s ol an i k üh n 1 1 0. 18 r hi zo ct on ia s pp . 2 2 0. 35 r hi zo pu s sp p. 1 3 15 6 5 4 2 36 6. 34 sc op ul ar io ps is b re vi ca ul is ( sa cc .) b ai n 1 1 0. 18 sc op ul ar io ps is b ru m pt i s al va ne td uv al 1 1 0. 18 sc yt al id iu m li gn ic ol a pe sa nt e 1 1 0. 18 sp ic ar ia e le ga ns c or da 3 2 5 0. 88 sp ic ar ia s im pl ic is si m a o ud em an s 1 1 2 0. 35 sp ic ar ia v io la ce a a bb ot t 3 6 2 2 13 2. 29 sp or ot ri ch um c ar ni s b ro ok s et h an sf or d 1 1 0. 18 to ru la h er ba ru m ( pe rs .) l in k ex f r. 1 1 0. 18 tr ic ho de rm a au re ov ir id e r if ai 1 1 1 3 0. 53 tr ic ho de rm a ha m at um ( b on .) b ai n 1 1 3 2 7 1. 23 tr ic ho de rm a ha rz ia nu m r if ai 1 1 0. 18 tr ic ho de rm a po ly sp or um ( l in k et p er s. ) r if ai 1 1 0. 18 n on -s po ru la ti ng fu ng i 1 1 1 1 6 1 4 1 16 2. 82 sp ri ng c ru ci fe ro us p la nt s 24 78 32 22 26 26 7 80 46 11 29 39 8 17 50 16 29 28 56 8 10 0. 0 su m o f p la nt 13 4 74 13 3 79 75 73 *1 19 99 2 2 00 0 3 20 01 190 b. majchrzak et al. genus rhizopus. ishimoto et al. (2000) confirmed the predominant role of the genus rhizopus in the rhizosphere of cruciferous plants. according to these authors, fungi of the genus rhizopus showed significantly higher tolerance for glucosinolates than fungi of the genus fusarium. conclusions 1. the largest and the most diverse fungal community was isolated from the soil environment of spring oilseed rape. 2. the fungal populations that colonized the rhizosphere and rhizoplane of spanish colewort and false flax were found to be the smallest. 3. members of the order mucorales dominated in the soil environment of cruciferous plants. 4. fungi of the genus fusarium were isolated least frequently from the soil environment of spanish colewort and most frequently from the soil environment of spring oilseed rape. references bojarczuk m., bojarczuk j. 1988. fitosanitarna ocena wartości przedplonów roślin zbożowych. fragm. agronom. 1(17): 5–23. bones a.m., rossiter j.t. 1996. the myrosinase-glucosinolate system, its organisation and biochemistry. physiol. plant. 97:194–208. charron c.s., sams c.e. 1999. inhibition of pythium ultimum and rhizoctonia solani by shredded leaves of brassica species. j. am. soc. hortic sci. 124:462–467. ishimoto h., fukushi y., yoshida t., tahara s. 2000. rhizopus and fusarium are selected as dominant fungal genera in rhizospheres of brassicaceae. journal of chemical ecology 26 (10): 2387–2399. kirkegaard j.a., sarwar m. 1998. biofumigation potential of brassicas. i. variation in glucosinolate profiles of diverse field-grown brassicas. plant soil 201:71–89. kurek e., kobus j. 1990. korzystne i szkodliwe oddziaływanie mikroflory ryzosferowej na wzrost i rozwój roślin. postępy mikrobiologii 29 (1/2): 103–123. majchrzak b., kurowski t.p., karpińska z. 2002. zdrowotność jarych roślin krzyżowych a grzyby zasiedlające ich nasiona. acta agrobot. 55 (1): 199–210. majchrzak b., okorski a., chodorowski b. 2004. zdrowotność korzeni i podstawy źdźbła pszenicy jarej uprawianej po różnych przedplonach. annales umcs 59(4): 1779–1788. majchrzak b., chodorowski b., okorski a. 2005. choroby podstawy źdźbła pszenicy ozimej uprawianej po roślinach przedplonowych z rodziny brassicaceae. acta agrobot. 59 (2): 307–318. majtahedi h., santo g.s., hang a.n., wilson j.h. 1991. suppression of root-knot nematode populations with selected rapeseed cultivars as green manure. j. nematol. 23:170–174. mańka k.. 1974. zbiorowiska grzybów jako kryterium oceny wpływu środowiska na choroby roślin. zesz. prob. post. nauk roln. 160: 9–22. marwar r., lodha s. 2002. brassica amendments and summer irrigation for the control of macrophomina phaseolina and fusarium oxysporum f. sp cumini in hot arid region. phytopathol. mediterr 41:45–54. morgan j. a. w., bending g. d., white p. j. 2005. biological costs and benefits to plant–microbe interactions in the rhizosphere. journal of experimental botany 56 (417): 1729–1739. oleszek w. 1997. glukozynolany występowanie i znaczenie ekologiczne. wiadomości bot. 39 (1/2): 49–58. patkowska e. 1998. wpływ resztek roślinnych na zdrowotność i plonowanie soi. roczn. akad. roln. poznań, ogrod. 27: 213-219. sarwar m., kirkegaard j.a., wong p.t.w., desmarchelier j.m. 1998. biofumigation potential of brassicas. iii. in vitro toxicity of isothiocyanates to soil-borne fungal pathogens. plant soil 201:103–112. fungi isolated from the rhizosphere 191 snapp s.s., date k.u., kirk w., o’neil k., kremen a., bird g. 2007. root, shoot tissues of brassica juncea and cereal secale promote potato health. plant soil. 294:55–72. sturz a. v., christie b. r. 2003. beneficial microbial allelopathies in the root zone: the management of soil quality and plant disease with rhizobacteria. soil & tillage research. 72: 107–123. grzyby ryzosferowe jarych roślin kapustnych s t r e s z c z e n i e w badaniach poddano analizie zbiorowiska grzybów ryzosferowych jarych roślin kapustnych. ryzosferę roślin kapustnych zasiedlały przede wszystkim grzyby z rzędu mucorales i rodzaju fusarium. w ryzoplanie roślin znacznie częściej występowali przedstawiciele rodzaju fusarium. korzenie rośliny kapustnych wydzielają do gleby glukozynolany, wtórne metabolity o właściwościach antygrzybowych, w ten sposób być może wpływają na zbiorowiska grzybów zasiedlających środowisko glebowe roślin. 2014-01-01t11:48:10+0100 polish botanical society goidanichiella sphaerospora, the world’s second record hanna kwaśna and jolanta behnke-borowczyk department of forest pathology, poznań university of life science wojska polskiego 71c, pl-60-625 poznań, kwasna@owl.up.pl kwaśna h., behnke-borowczyk j.: goidanichiella sphaerospora, the world’s second record. acta mycol. 45 (1): 33–35, 2010. a specimen of goidanichiella sphaerospora was found in pinus sylvestris forest soil in międzychód, northwestern poland (52.601, 15.889883), in october 2009. this is the world’s second record of g. sphaerospora. bimorphic conidial heads and conidia are reported for the first time. goidanichiella sphaerospora forms aspergillusand penicillium-like conidial heads. conidia formed at +24oc are oval to ellipsoidal, often apiculate, smooth, rather thick-walled, hyaline, with one oil drop inside, 3-4 × 2-3 μm. additional ellipsoidal to cylindrical, thinwalled, 4-6.5 (-8) × 2-3.0 μm conidia are formed only after incubation for at least 7 days at +4oc in darkness. key words: goidanichiella sphaerospora, morphology, taxonomy introduction during a study of biodiversity in fungal communities in forest soils treated with different methods of post-harvest utilization of wood debris and pre-planting preparation of soil, goidanichiella sphaerospora matsushima was identified on the basis of its morphology in vitro. the fungus produced numerous, characteristic, brownpigmented conidiophores which ended mostly with aspergillus-like radiate, fertile conidial heads. conidia were cohering in glistening white slimy globule heads. four species of goidanichiella g.l. barron ex w. gams are currently known: g. barronii w. gams, steiman & seigle-murandi, g. cylindrospora d.w. li & g.h. zhao, g. fusiformis k.d. hyde, yanna, pinnoi & e.b.g. jones and g. sphaerospora (barron 1968; matsushima 1975; gams et al. 1990, 2009; hyde et al. 2002; li, zhao 2007). goidanichiella sphaerospora has previously been found only once, in forest soil in hokkaido, japan, by matsushima (1975). the aim of this paper is: (i) to present the world’s second record of g. sphaerospora, (ii) to report newly observed morphological characteristics (figs 1–8). acta mycologica vol. 45 (1): 33–35 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 34 h. kwaśna and j. behnke-borowczyk materials and methods the fungus was isolated using the soil plate method (warcup 1950). conidiophores and conidia of the fungus grown on potato dextrose agar (pda; 20 g difco pda, 20 g agar in 1 l of distilled water) and synthetic nutrient agar (sna; 1 g kh2po4, 1 g kno3, 0.5 g mgso4·7 h2o, 0.5 g kcl, 0.2 g glucose, 0.2 g sucrose, 20 g agar, 1 l distilled water) were mounted in water. microscopic observations were made using nomarski differential interference contrast optics. results a single specimen of g. sphaerospora was found in p. sylvestris forest soil in międzychód, northwestern poland (52.601, 15.889883), in october 2009. the soil had been deep ploughed in 2005, when the 10-15 cm deep, humus-rich soil layer was inverted. the post-harvest wood debris had remained on the soil surface since 2005, usually between rows along which 6-year-old p. sylvestris trees were planted. the isolate of g. sphaerospora on pda formed a thin colony, at first hyaline, later greyish, particularly in the centre, spreading broadly, with daily radial increments of 13-15 mm at 24°c. conidiophores were numerous, scattered, erect, pale brown, with 4 (-5) septa (including the one at the base), 150-400 (-550) μm high, 15-20 μm diameter at the base, tapering to 8-10 μm diameter near the tip, supported by a swollen, more or less cylindrical, horizontal, brown cell, about 60-70 × 8-15 μm, present in the subtending hypha. most conidiophores ended with aspergillus-like, biseriate conidial heads. their vesicles, often with a slight constriction below the inflation, were up to 40 μm diameter, fertile usually only in the upper part, bearing a series of moderately divergent or strictly parallel metulae. each metula was bearing 3-5 appressed tapering phialides. metulae of the aspergillus-like conidial heads were cylindrical to obconical, subhyaline, (4.8-) 6-7 × 3.6-4.8 μm. phialides were cylindrical to conical, hyaline, 4.8-7.2 × 2.4-3.6 μm. rarely, more often on pda, conidiophores that were usually shorter and always hyaline with penicillium-like conidial heads appeared. they had none or less pronounced vesicles and 4-10 parallel metulae, each bearing 2-5 phialides. conidia, oval to ellipsoidal, often apiculate, smooth, rather thick-walled, hyaline, with one oil drop inside, 3-4 × 2-3 μm, were formed on both the aspergillusand penicillium-like conidial heads on sna and pda at 24°c. ellipsoidal to cylindrical, thin-walled, 4-6.5 (-8) × 2-3.0 μm conidia were formed after incubation for at least 7 days at +4oc in darkness. minimum, optimum and maximum temperatures for growth were 10°c, 25°c and 35°c. the fungus was deposited in cabi bioscience, uk centre (imi 398538) and centraalbureau voor schimmelcultures, utrecht, the netherlands (cbs 127247). the its1/2 dna sequence is in european nucleotide archive under embl no fr681846. goidanichiella sphaerospora 35 discussion there are only two goidanichiella species with biseriate vesicles, viz. g. barronii and g. sphaerospora. the latter was separated from g. barronii on the basis of: (i) width of conidiophores, which in g. barronii are (6-)8-13 μm near the base and 4-7.5 μm near the tip, (ii) size of metulae and phialides, which in g. barronii are 6.5-11 × 2-3.5 μm and 7.5-10 × 1.5-2.5 μm, respectively, (iii) formation of monomorphic conidia at 24oc in a day-night cycle, (iv) formation of bimorphic conidia only at 4oc in darkness. the ellipsoidal to cylindrical, thin-walled, 4-6.5 (-8) × 2-3.0 μm conidia, formed only at low temperature and in darkness, resembled the similar allantoid, thin-walled, 4-6.5(-7) × 1.4-2.0 μm conidia produced on conidiophores with less pronounced vesicles by g. barronii (gams et al. 1990). our isolate of g. spaherospora differed from the original isolate described by matsushima (1975) in: (i) formation of more slender metulae and phialides of the aspergillus-like conidial heads, (ii) longer persistence of conidial structures which do not collapse after maturity, (iii) absence of globose conidia reported by matsushima (1975), (iv) formation of larger, ellipsoidal to cylindrical conidia at low temperature and in darkness. references barron g. l. 1968. the genera of hyphomycetes from soil. william and wilkins, baltimore. gams w., steiman r., seigle-murandi f. 1990. the hyphomycete genus goidanichiella. mycotaxon 38: 149–159. gams w., seifert k. a., morgan-jones g. 2009. new and validated hyphomycete taxa to resolve nomenclatural and taxonomic issues. mycotaxon 110: 89–108. hyde k. d., yanna, pinnoi a., jones e. b. g. 2002. goidanichiella fusiforma sp. nov. from palm fronds in brunei and thailand. fung. diver. 11: 119–122. li d. w., zhao g. 2007. goidanichiella cylindrospora sp. nov. from connecticut, usa. mycotaxon 101: 41–45. matsushima t. 1975. icones microfungorum a matsushima lectorum. kobe. 77–78. warcup j. h. 1950. the soil plate method for isolation of fungi from soil. nature 166: 117, london. goidanichiella sphaerospora, drugie stanowisko na świecie streszczenie goidanichiella sphaerospora znaleziono w glebie leśnej, w młodniku pinus sylvestris, w międzychodzie (polska północna, 52.601, 15.889883), w październiku 2009. jest to drugie stanowisko tego grzyba na świecie. poprzednio grzyba stwierdzono w glebie leśnej w japonii. grzyb tworzy zarodniki konidialne na bimorficznych konidioforach przypominających konidiofory aspergillus i penicillium. wystepują dwa rodzaje zarodników. w temperaturze 24oc, na pda i sna, tworzą się konidia, które są owalne do eliptycznych, ostro zakończone u podstawy, gładkie, grubościenne, hialinowe, z jedną kroplą wewnątrz, 3-4 × 2-3 μm. w temperaturze 4oc, przy braku światła, tworzą się dodatkowe konidia, które są eliptyczne do cylindrycznych, cienkościenne, 4-6,5 (-8) × 2-3,0 μm. 2014-01-01t11:50:14+0100 polish botanical society additions to the biota of lichenized fungi of poland adam flakus1 and martin kukwa2 1laboratory of lichenology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, a.flakus@botany.pl 2department of plant taxonomy and nature protection, university of gdańsk al. legionów 9, pl-80-441 gdańsk, dokmak@univ.gda.pl flakus a., kukwa m.: additions to the biota of lichenized fungi of poland. acta mycol. 44 (2): 249–257, 2009. new records of five lichenized fungi from poland are provided. hypotrachyna afrorevoluta, lecanora quercicola, rhizocarpon superficiale and strigula ziziphi are new to poland. of these, strigula ziziphi is reported also as new to central europe and hypotrachyna afrorevoluta as new to the carpathians. additionally, thelenella muscorum var. octospora is recorded from its second locality in poland as new to the polish carpathians. hypostictic acid chemosyndrome has been noticed for the first time in european (poland) and south american (bolivia) populations of rhizocarpon superficiale. key words: chemotaxonomy, neglected lichens, hypotrachyna, lecanora, rhizocarpon, strigula, thelenella introduction currently known lichen biota of poland include about 1600 lichens (fałtynowicz 2003) and 222 lichenicolous fungi and myxomycetes (czyżewska, kukwa 2009), and can be considered as rather rich and well investigated. however, the country is also still non-uniformly investigated, and several additional species, including taxa new to science, can be found, what has been recently demonstrated by several authors (cykowska, flakus 2005; kukwa, diederich 2005; czarnota 2007; flakus 2007; kukwa, kubiak 2007; czarnota, kukwa 2008; kossowska 2008; motiejūnaitė, czyżewska 2008; motiejūnaitė, kukwa 2008; kukwa, jabłońska 2009; zhurbenko et al. 2009). in the paper we present further four records of lichenized fungi new to poland, of which strigula ziziphi is new to central europe, and hypotrachyna afrorevoluta and thelenella muscorum var. octospora are respectively new to the carpathians and the polish carpathians. additionally hypostictic acid is reported for the first time from european and south american specimens of rhizocarpon superficiale. acta mycologica vol. 44 (2): 249–257 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 250 a. flakus and m. kukwa material and methods the present study is based on the specimens deposited in following lichen herbaria: b, kram, lg, lpb, ugda. the anatomy and morphology of the material were studied by traditional tools used in lichenology. secondary metabolites were identified by thin layer chromatography (tlc) in solvents a, b and/or c according to the methods of orange et al. (2001). localities of all polish examined specimens are mapped according to the modified atpol grid square system (cieśliński, fałtynowicz 1993; kukwa et al. 2002). results and discussion hypotrachyna afrorevoluta (krog & swinscow) krog & swinscow, lichenologist 19: 420 (1987). syn. parmelia afrorevoluta krog & swinscow, norweg. j. bot. 26: 22 (1979); parmelinopsis afrorevoluta (krog & swinscow) elix & hale, mycotaxon 29: 242 (1987). morphology and chemistry. the species has foliose, grey or grey-green, rather compact thallus, up to ca 4 in diam., lobes are rounded, separate, but more or less overlapping towards the center, with smooth surface, and sparsely ciliate margin. soredia are produced in laminal or submarginal soralia formed from pustules. lower surface is usually shiny and brown, dark brown to almost black up to the margin. the species produces atranorin in the cortex and gyrophoric (major) and lecanoric (minor) acids together with related accessory substances (in trace amounts) in medulla (swinscow, krog 1988; ertz et al. 2008). the cortex is k+ yellow, whereas the medulla reacts c+ pink-red and kc+ red. notes. h. afrorevoluta is very similar to h. revoluta (flörke) hale, and all polish herbarium specimens were found under the latter name. both taxa are sorediate and produce the same lichen substances, but they differ mainly in the formation of soralia. in h. afrorevoluta they are formed from laminal or submarginal pustules, which break and start to produce granular soredia, whereas in h. revoluta soralia develop apically on the upper surface of relatively large and elongated lobes, and soredia are more farinose than in h. afrorevoluta. according to ertz et al. (2008) they can be also distinguished by the colour of lower surface of young lobes; it is brown to dark brown or blackish and shiny in h. afrorevoluta, whereas in h. revoluta it is usually pale brown and matt. in so far studied polish material the soralia were well developed in few specimens of both taxa, and thus the determination of them was not difficult. some thalli were covered with dust or algae, deformed, degenerated and probably collected from polluted or disturbed ecosystems, and then soralia were often badly developed (i.e. some produced at the apices, but some formed also laminally, or very few young soralia were present). in such cases, we used the colour of lower cortex for the determination of species, and in these samples it was rather pale brown at the margin of lobes, and thus suggesting h. revoluta; however it was not matt, but shiny. we attributed additions to the biota 251 those specimens to h. revoluta, but with a hesitation. their true identity may become more clear when the morphology of all polish collections is studied. it was already noted by ertz et al. (2008), that specimens can not be always determined. habitat. the species is mainly corticolous on bark of many tree species (swinscow, krog 1988; ertz et al. 2008), but in poland one specimen was also found on rock. two specimens from northern poland were found in humid forests, but habitat details were missing on remaining labels. general distribution. h. afrorevoluta is widely distributed in the world. originally it was described from africa (krog, swinscow 1979; swinscow, krog 1988), but later it was reported from belgium, france, british isles, germany, luxembourg, the netherlands, norway, portugal and switzerland in europe (santesson 1993; masson 2005; hitch 2007; spier et al. 2007; dolnik et al. 2008; ertz et al. 2008), china in asia (chen et al. 2003), north america (knudsen, lendemer 2005; lendemer 2006) and south america (adler, elix 1992). distribution in poland. h. afrorevoluta is reported here for the first time from poland and the carpathians. it can be rather widely distributed in the country as localities of this lichen are known from northern and southern part of poland. possibly it can be more common than h. revoluta, similar as in belgium, northern france, luxembourg and the netherlands (spier et al. 2007; ertz et al. 2008). specimens examinated. poland. ac 34 – wybrzeże słowińskie coast, mierzeja sarbska split, forest section no. 14dm, black alder forest, on alnus glutinosa, 4 oct. 1998, leg. w. fałtynowicz (ugda-l 13353). bd 53 – pojezierze iławskie lakeland, e of szadowo village, liwa river valley, n bank of the river, 53°46’20”n, 19°03’28”e, hornbeam-lime forest, on alnus glutinosa, 16 april 2006, leg. m. kukwa 4946 (ugda-l 13528). fg 01 – roztocze środkowe, kosobudy forest inspectorate, forest district czerkies, ca 50°35’n, 23°01’e, the edge of the forest, on fagus sylvatica, 30 june 1952, leg. z. tobolewski (ugda-l 4480). gf 10 – beskid niski mts., diabli kamień rock near folusz village, on rock, ca 1954, leg. t. sulma (ugda-l 12155). lecanora quercicola coppins & p. james, lichenologist 11: 145 (1979). morphology and chemistry. l. quercicola belongs to l. saligna-group, and is characterized by apothecia with a pale yellowish, corticate, lecanorine margin, pale yellowish, to brown and ± pruinose disc, yellow-brown to olive brownish epihymenium not reacting with n, ellipsoid ascospores (8.6–10 × 4.3–4.8 μm), weakly curved, luniform, non-septate macroconidia measuring 8.5–9.5 × (2.5–)2.7–3.0(–3.2) μm, and the production of isousnic acid as a major secondary metabolite (coppins, james 1979; boom, brand 2008). polish specimen matches this characteristic well, except the small difference in the size of macroconidia, which are (8–)8.5–9 × 2.5–3.0 μm. notes. l. quercicola is very similar to l. saligna (schrad.) zahlbr. s.str. both taxa have almost the same morphology, size of ascospores, and chemistry, but they predominantly differ in dimensions of macroconidia; they are 6.0–8.1 × 2.0–2.4 μm in l. saligna, whereas 8.5–9.5 × 2.7–3.0 μm in l. quercicola (boom, brand 2008). when we revised the material of both taxa from ugda, we found, that pycnidia containing macroconidia were usually difficult to find on most thalli. pycnidia are very small and often pale brown, and they can be mistaken for very young apothecia or dust impregnation. sometimes, pycnidia with microconidia were much more abundant than those with macroconidia. therefore, several squash preparations are advised to be made for the identification of single specimen. some specimens did 252 a. flakus and m. kukwa not posses conidiomata with macroconidia at all, and such material could not have been determined. habitat. l. quercicola grows on wood and bark of trees (quercus spp. and olea europaea) in open places in woodlands (coppins, james 1979; boom, brand 2008). in poland it was found on wood of fallen decorticated branch (probably salix acutifolia) on grey sand dune. the place was well lit and sheltered from wind. general distribution. the species is known only from europe and so far it has been reported from france, germany, great britain, portugal and spain (coppins, james 1979; boom, brand 2008). distribution in poland. l. quercicola is here reported as new to poland. until know it has been found only by the sea shore in one locality during the inventory of the lichen biota of ‘helskie wydmy’ nature reserve. the revision of material l. saligna-group deposited in ugda (ca 30 vouchers) did not yield any additional specimen of l. quercicola. specimen examinated. poland. ad 51 – mierzeja helska split, ‘helskie wydmy’ nature reserve, jastarnia forest division, 54°39’18”n, 18°46’42”e, sand dunes, on wood of fallen tree branch, 12 may 2009, leg. m. kukwa 7388 & i. s. stepanchikova (ugda). rhizocarpon superficiale (schaer.) vain., acta. soc. fauna fl. fenn. 53 (1): 319 (1922). syn. lecidea superficiale schaer., lich. helvet. spicil. [part 3]: 125 (1822). morphology and chemistry. r. superficiale is characterized by crustose, areolate, epruinose thallus, and usually well developed black prothallus. areoles are bright yellow to whitish yellow, with smooth or often scabrose surface. medulla is white and reacts i–, k+ yellow, pd+ orange. the species produces rhizocarpic, hypostictic, stictic, and/or norstictic acids (geyer et al. 1984; feuerer, timdal 2004). apothecia are black, epruinose, usually angular, plane to weakly convex, 0.5–1(2) mm in diam., with black, often persistent margin. asci are 8-spored, and ascospores are 1-septate, dark green-brown when mature, 14–18 × 6–8 μm (runemark 1956; wirth 1995; feuerer, timdal 2004; fletcher et al. 2009). notes. r. superficiale can be easily distinguished from other yellow, morphologically similar species by 1-septate, small ascospores and the presence of norstictic, stictic or hypostictic acids as major secondary compounds (wirth 1995; feuerer, timdal 2004; fletcher et al. 2009). this species is diverse in regards of chemical compounds. in european populations either stictic or norstictic acid have been recorded as a main substances, whereas in north american material hypostictic and stictic acids, with norstictic acid as a minor compound (geyer et al. 1984; wirth 1995; feuerer, timdal 2004). in specimens examined by us from poland and bolivia hypostictic and stictic acids were detected as a major compounds, together with related additional substance in small amount (possibly cryptostictic acid). it is the first record of this substance from european and south american populations of r. superficiale. the norstictic acid was not confirmed from polish specimens neither by both tlc analysis and microchemical reactions under the microscope with koh solution (characteristic red, needle-shaped crystals were not observed). habitat. the species occurs on siliceous, exposed rocks in high mountains or cool areas (runemark 1956; wirth 1995; feuerer, timdal 2004; fletcher et al. 2009). additions to the biota 253 three polish specimens were collected from vertical, south exposed granite rock in the subnival belt. general distribution. the species is a bipolar, widespread species, known from africa, antarctica, asia, australasia, europe, north america and south america (runemark 1956; feuerer, timdal 2004; olech 2004; lisická 2005; fletcher et al. 2009). distribution in poland. r. superficiale is reported in this paper for the first time from poland. so far it has been found only in two localities in tatra mts. at the altitude above 2300 m. specimens examinated. poland. ge 60 – western carpathians, high tatra mts., mięguszowiecki szczyt mt., 49°11’13”n, 20°03’34”e, 2438 m, subnival belt, on vertical granite rock, 9-17 aug. 2003, leg. a. flakus 1500, 2007 (kram); pośredni mięguszowiecki szczyt mt., 49°11’04”n, 20°03’50”e, alt. 2360 m, subnival belt, on vertical granite rock, 3 aug. 2003, leg. a. flakus 1213 (kram). additional specimens examinated. bolivia. dept. la paz, prov. murillo, upper part of valley under mt. chockaltaya, 16°20’41”s, 68°08’10”e, alt. 4980 m, on siliceous rock, 9 dec. 2004, leg. a. flakus 4461 (b, kram, lpb). strigula ziziphi (a. massal.) cl. roux & sérus., biblioth. lichenol. 90: 55 (2004). syn. sagedia ziziphi a. massal., miscellanea lichenologica: 30 (1856); strigula mediterranea etayo, lichenologist 25: 258 (1993). morphology. s. ziziphi has thin, in most parts endophloedic thallus in shades of brown, trentepohlia as photobiont, black perithecia (0.2–)0.3–0.4(–0.5) mm in diam., hamathecium consisting of mostly simple, only rarely branched and anastomosing paraphysoids and claviform asci with biseriate spores; spores are 1-septate, distinctly constricted at the septa and early breaking into two parts, specially at the ascus dehiscence, and measure (18–)19.5–27(28) μm. the species develops two types of conidia, macroand microconidia, which are produced in macroand micropycnidia, respectively. macroconidia are 1-septate, guttulate, often with gelatinous (mucoid) appendages, and measure (8–)9–11(–11.5) × (2.5–)3–4(–4.5) μm; macropycnidia are 0.12–0.2(–0.3) mm in diam. microconidia are fusiform or subfusiform, 2.5–5 × 1–2 μm; micropycnidia are 0.1–0.18 mm (etayo 1993, sub s. mediterranea; roux, sérusiaux 2004). in polish material only macropycnidia with macroconidia were found. notes. so far this species has not been found with perithecia in poland, and therefore it can be very easily overlooked in the country; difficulties may also appear in the identification of such material; however up to our knowledge, there is no other corticolous lichen species with 1-septate macroconidia as described above and trentepohlia as photobiont. s. affinis (a. massal.) r. c. harris is the only other corticolous member of the genus known from poland (fałtynowicz 2003). it can also develop sterile thalli with macropycnidia, but it differs in 3-septate macroconidia (roux, sérusiaux 2004). in poland s. ziziphi can be also confused with a common anisomeridium polypori (ellis & everh.) m. e. barr and rare a. ranunculosporum (coppins & p. james) coppins [syn. arthopyrenia ranunculospora coppins & p. james], arthopyrenia cerasi (schrad.) a. massal. and a. cinereopruinosa (schaer.) körb. they all differ in pycnidial and conidial characters. anisomeridium polypori has conical macropycnidia, smaller macroconidia (3.5–4.5 × 1.8–3 μm) extruded as a white cirrus from ostiolum; micropycnidia are ± immersed, globose and up to 0.1 mm in diam., with small, 2–3 × 1–1.3 μm microconidia (coppins et al. 2009). in a. ranunculosporum 254 a. flakus and m. kukwa macropycnidia and micropycnidia are smaller (respectively up to 0.12 mm, and up 0.06 mm in diam.), macroconidia have similar size to those of strigula ziziphi, but are predominantly non-septate, and microconidia are bacilliform and narrower (0.5–0.8 μm in width) (coppins et al. 2009). both, arthopyrenia cerasi and a. cinereopruinosa are not lichenized. a. cerasi has 3-septate macroconidia, which are longer (11–13 μm), but narrower (2–2.5 μm) than in s. ziziphii; micronidia are much longer and narrower (9–14 × 0.8 μm). in a. cinereopruinosa macroconidia are simple (8–11 × 1.8–2.2 μm) and microconidia mostly longer and wider (4.7–6 × 1 μm) (coppins, orange 2009). habitat. the species s. ziziphi grows on different types of trees, and at least in iberian peninsula it is photophilous and somewhat nitrophilous lichen growing in rather humid places (etayo 1993; roux, sérusiaux 2004). in poland it was found on bark of willow in open, but humid place by the river. general distribution. the species has been so far reported from croatia, france, greece, italy, the netherlands, portugal, switzerland, ukraine (crimean peninsula), and outside europe only in canary islands (etayo 1993, sub s. mediterranea; scheidegger et al. 2002, sub s. mediterranea; roux, sérusiaux 2004). distribution in poland. s. ziziphi it is reported for the first time from poland and central europe. its finding in this regions appears rather unexpected, but the species can develop sterile thalli with macropycnidia only (as in polish material), therefore it can be easily overlooked and its true distribution may have not yet been determined. specimen examinated. poland. ac 51 – wybrzeże słowińskie coast, smołdzino village, by łupawa river, on salix sp., 19 march 1995, leg. w. fałtynowicz, det. e. sérusiaux (ugda-l 10053, dupl. lg). thelenella muscorum var. octospora (nyl.) coppins & fryday, lichenologist 36 (2): 91 (2004). syn. verrucaria muscicola var. octospora nyl. in ohlert, schriften königl. phys.-ökon. ges. königsberg 11: 43 (1870); chromatochlamys muscorum var. octospora (nyl.) h. mayrhofer & poelt, herzogia 7 (1-2): 37 (1985). morphology. t. muscorum var. octospora is a terricolous or epiphytic species, characterized by thin, whitish or pale brown crustose thallus, with chlorococcoid photobiont, and small, rounded or broadly pyriform, dark brown perithecia, ussually immersed in substratum, 0.3–0.6 mm in diam. it has persistent, branched and anastomozed paraphysoids, functionally bitunicate, thick walled, thelenella-like asci, with 6–8 ascospores; ascospores are thin-walled, muriform, elongate-ellipsoid and colourless, ca 40–60 × 12–20 μm (mayrhofer, poelt 1985; fryday, coppins 2004; orange et al. 2009). notes. t. muscorum var. octospora is very similar to t. muscorum var. muscorum, which differs by 2–4-spored asci and bigger ascospores (60–110 × 20–27 μm) (mayrhofer, poelt 1985). t. muscorum var. octospora can be also confused with strigula confusa fryday, coppins & comminis, superficially similar species that was recently described from british isles, but the latter taxon differs in black, globose perithecia, other ascus type, trentepohlia photobiont and smaller ascospores (fryday, coppins 2004). habitat. this lichen occurs mainly on bryophytes, plant debris, basic-barked trees and rarely on soil, basic rocks or lichen thalli (mayrhofer, poelt 1985; fryday, coppins 2004; orange et al. 2009). in poland it was discovered on plant debris in mylonitized area of the subnival belt in the tatra mts. additions to the biota 255 general distribution. t. muscorum var. octospora is known from british isles, denmark, germany, fennoscandia (sweden and norway), greenland and north america (mayrhofer, poelt 1985; alstrup 1993, 2004; santesson et al. 2004; esslinger 2009). distribution in poland. the taxon was originally described in 1870 as verrucaria muscicola var. octospora from northern poland (mayrhofer, poelt 1985), however, it has not been included in the last checklist of lichen biota. here we report its second polish locality from tatra mts., which is also the first record for the polish carpathians. specimen examinated. poland. ge 60 – szpiglasowa przełęcz pass, 49°11’53”n, 20°02’34”e, 2105 m, subnival belt, on plant debris in mylonitized area, 24 aug. 2004, leg. a. flakus 3376 (kram). acknowledgement. we are very grateful to professor emmanuël sérusiaux (liège) for the determination of strigula ziziphi. it is our pleasure and honor to dedicate this paper to professor krystyna czyżewska (łódź) on the occasion of her anniversary. references adler m. t., elix j. a. 1992. new records of hypotrachyna and parmelinopsis lichens (ascomycotina, parmeliaceae) from northwest and central argentina. mycotaxon 43: 283–288. alstrup v. 1993. news on lichens and lichenicolous fungi from the nordic countries. graphis scripta 5 (2): 96–104. alstrup v. 2004. new records in distribution of lichens and lichenicolous fungi. graphis scripta 16 (2): 46–57. boom van den p., brand 2008. some new lecanora species from western and central europe, belonging to the l. saligna group, with notes on related species. lichenologist 40 (6): 465–497. chen j.-b., wang s. l., elix j. a. 2003. parmeliaceae (ascomycota) lichens in china’s mainland i. the genera canomaculina, parmelina, parmelinella and parmelinopsis. mycotaxon 86: 19–29. cieśliński s., fałtynowicz w. 1993. note from editors. 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substancja ta znana była dotychczas jedynie z północnoamerykańskich okazów tego gatunku. 2014-01-01t11:49:46+0100 polish botanical society the role of wild boars in spore dispersal of hypogeous fungi federica piattoni1, francesca ori2, marco morara3, mirco iotti4 and alessandra zambonelli5 dipartimento di scienze agrarie, viale fanin 46, i-40127 bologna 1federica.piattoni@unibo.it, 2francesca.ori4@studio.unibo.it, 3mamo46it@yahoo.it, 4mirco.iotti2@unibo.it, 5corresponding author: alessandr.zambonelli@unibo.it piattoni f., ori f., morara m., iotti m., zambonelli a.: the role of wild boars in spore dispersal of hypogeous fungi. acta mycol. 47 (2): 145–153, 2012. wild boars (sus scrofa l.) are well-known for soil disturbance in natural and cultivated truffières but their role in spore dispersal is poorly investigated. in the present work we studied the occurrence of hypogeous fungal spores in faecal contents of 14 wild boars randomly hunted in “parco dei gessi e calanchi dell’abbadessa” regional park (north of italy) where truffle production has been previously investigated for three years. six methods for spore analysis in faeces were compared and the suspension of faeces in znso4 (70%) solution resulted to be the most reliable. hypogeous fungal spores, including tuber magnatum and tuber aestivum spores, were detected in 9 animals. this result suggests that the detection of fungal spores in faeces of wild boars may provide information on the presence of hypogeous fungi in an area. however, the poor abundance of spores suggests that the wild boar can be considered an opportunistic mycophagist, ingesting truffles only occasionally, as a seasonal source of food. considering the magnitude of wild boar movements during seasonal migrations, it is possible to speculate that they play a key role in truffle long distance dispersal. key words: sus scrofa, truffles, mycophagous, spore dispersal, faeces introduction hypogeous fungi comprise species belonging to several genera of ascomycota (true truffles), basidiomycota and muromycotina (false truffles) (trappe et al. 2009; bonito et al. 2013). the hypogeous lifestyle comported several morphological changes, as the loss of the ability to discard spores actively (trappe, claridge 2005). this led to develop several survival strategies, as the development of a strong aroma, even typical of any acta mycologica vol. 47 (2): 145–153 2012 146 f. piattoni et al. truffle species (gioacchini et al. 2005). this aroma is useful to the fungus to be found and eaten by wild animals, mainly rodents and mammals, ensuring spore dispersal into the environment by animal faeces (cazares et al. 1999; trappe, claridge 2005). indeed, faeces examination of mycophagous rodents, such as the giant whitetailed rat, uromys caudimaculatus krefft, the golden mantled ground squirrel, spermophilus saturates rhoads, and the deer mouse, peromyscus maniculatus wagner, demonstrated that truffles represent a consistent part of their diet (comport, hume 1998; cork, kenagy 1989). among mammals, wild boars (sus scrofa l.) are sadly known to damage truffle production in natural and cultivated truffières (ricci 2008; moreno-arroyo et al. 2005). the negative impact of wild boars on truffles is attributed to soil disturbance and ascoma consumption, as we demonstrated for tuber aestivum in natural truffières in central italy (salerni et al. 2011). however, less is known on the possible ecological role of wild boars in truffle spore dispersal (genard et al. 1986; steiner, fielitz 2009). as far as we know, only the former authors reported specifically about t. aestivum consumption by wild boars, but the paper lacks in technical information about spore isolation methods. thus, in the present work we investigated the occurrence of hypogeous fungal spores in faecal contents of wild boars hunted in “parco dei gessi e calanchi dell’abbadessa” regional park, an area known for truffle production. materials and methods study area. the study was carried out in “parco dei gessi bolognesi e calanchi dell’abbadessa” (ente di gestione per i parchi e la biodiversità emilia orientale, 2013) which is located in the south eastern hilly area of bologna (central italy) in the municipalities of bologna, ozzano dell’emilia, pianoro and san lazzaro di savena (surface area 4.815,87 ha). in this area, the forests are mainly represented by ostrya carpinifolia scop. and quercus pubescens willd. (all. orno osrtryon auct. ital. and all. ostryo-carpinion orientalis, horvat 1954) (corbetta 1994). the park is characterized by a temperate mediterranean climate (köppen-geiger classification), where the highest and lowest temperatures occur in july-august and december-january, respectively, and the annual average precipitation is 750 mm (pieri et al. 2011). truffles, in particular t. aestivum and t. magnatum, are principally found in q. pubescens mixed forests, in calcareous soils derived from marnous arenaceous rocks, within the municipalities of ozzano dell’emilia, pianoro and san lazzaro di savena. hypogeous sporoma collection. fruiting bodies of truffles were collected in q. pubescens forests of the park, using trained dogs, between september and january 2004-2007. animals. between october 3rd and december 4th 2011, the faeces of 14 wild boars hunted in the park area, in compliance with the permitted hunter-kill ratio actions established by bologna province, were examined. animals, randomly chosen regardless sex, age and weight, were dissected by the “azienda agricola s. uberto” accredited slaughtering house (monterenzio, bologna) (provincia di bologna 2007). wild boars and truffle spores 147 preparation of faecal samples for spore detection. faeces were forced out of the rectum and put into a screw cap vial. in the lab, faecal samples were weighted and diluted 10-fold with sterile distilled water. the suspension was decanted for one hour. the precipitate was sifted through a series of metal sieves of decreasing mesh size (800, 400, 150, 60 and 20 μm). only the material between 150 μm and 20 μm was in size considered for centrifugation (1500 rpm, 3 min) with sterile distilled water because hypogeous fungal spores (and asci) generally range between these dimensions. the supernatant was discharged and the precipitate was treated with 6 modified solutions: 1) nacl (26.5%); 2) mgso4 (35%); znso4 (33%); znso4 (70%); 5) sucrose (68%); 6) sucrose gradient (crede 2007; gudmundsdottir, skirnisson 2006; pet informet 2008; mitosciences 2007). for the latter, 3 different sucrose solutions (35%, 25% and 15%) were sequentially and carefully layered in a 15 ml tube and, finally, 1 ml of faecal precipitate was layered on top. after sucrose gradient centrifugation (1500 rpm, 2 min), three aliquots (100 μl each) were collected from each 1-ml interval of the gradient, transferred onto a slide and immediately examined. for the first 5 methods, the faecal precipitate (10 g) was 5-fold diluted with the corresponding solution and centrifuged (1500 rpm, 3min). after centrifugation, new solution was added to the very top of the tube and a cover slip was placed on it for 10-15 min for recovering the floating fungal spores. cover slips were mounted on a slide and immediately examined under a light microscope. each sample was treated in triplicate. among the six methods tested, the znso4 (70%) solution resulted to be the most reliable to isolate spores and thus it was used for analyzing all faecal samples. morphological identification. fresh samples of fruiting bodies were preliminary identified on the basis of their macroscopic (colour, surface, smell, etc.), and microscopic characteristics (morphological and biometric characteristics of spores and peridium cells) numbered, dried and stored in the herbarium of the dipartimento di scienze agrarie (cmi-unibo), university of bologna (italy). the spores found in the faeces were identified basing on their external characteristics (shape, dimensions and type of ornamentation). tuberkey (zambonelli et al. 2000) was used as reference for tuber spp. identification whereas montecchi and sarasini (2000) monography was used as reference for the species of hypoeous fungi belonging to different genera of ascomycetes and basidiomycetes. results hypogeous sporoma collection. during the surveys, only a few ascomata of t. aestivum and t. magnatum were found because the area is regularly visited by other truffle hunters, although truffle harvesting within the park is forbidden. other hypogeous ascomycetes were common in the park, such as t. excavatum, t. rufum, t. macrosporum t. borchii, t. dryophilum, t. brumale, balsamia vulgaris, stephensia bomycina, genea spp. basidiomycetes were only represented by the species of the hymenogaster genus and by melanogaster ambiguus (tab. 1). 148 f. piattoni et al. detection and identification of faecal spores. the data of the 14 wild boars examined and the characteristics of the truffle spores detected in the corresponding faecal samples are shown in table 2. no fungal spores were detected in 5 animals whereas in 9 ones spores of hypogeous fungi, including t. magnatum and t. aestivum, were identified (fig. 1). other unidentified fungal spores were present in most faecal contents, including several spores of alternaria spp. (data not shown). truffle spores were found in both sexes and in the animals from all the municipalities within the park. table 1 hypogeous fungi found in “parco dei gessi bolognesi e calanchi dell’abbadessa” regional park (bologna, italy) herbarium n. species date municipality 3382 balsamia vulgaris vittad. 30 12 2006 san lazzaro 3352 23 01 2007 ozzano dell’emilia 3357 genea fragrans (wallr.) sacc. 23 01 2007 ozzano dell’emilia 2546 15 11 2004 san lazzaro 3205 08 11 2006 ozzano dell’emilia 3375 30 12 2006 san lazzaro 3372 03 12 2006 san lazzaro 3351 genea lespiaultii corda 23 01 2007 ozzano dell’emilia 3373 genea verrucosa vittad. 03 12 2006 san lazzaro 2466 hymenogaster lycoperdineus vittad. 27 09 2004 san lazzaro 3343 24 01 2007 ozzano dell’emilia 3353 melanogaster ambiguus (vittad.) tul. & c. tul. 23 01 2007 ozzano dell’emilia 3379 30 02 2007 san lazzaro 3344 24 01 2007 ozzano dell’emilia 3867 27 12 2005 san lazzaro 2552 15 11 2004 san lazzaro 2550 15 11 2004 san lazzaro 2452 11 09 2004 pianoro 1099 25 09 2007 san lazzaro 2449 tuber aestivum vittad. 11 09 2004 pianoro 3192 08 11 2006 ozzano dell’emilia 3380 30 12 2006 san lazzaro 3348 tuber borchii vittad. 23 01 2007 ozzano dell’emilia 3346 23 01 2007 ozzano dell’emilia 2544 tuber brumale vittad. 15 11 2004 s. lazzaro 3354 tuber dryophilum tul. & c. tul. 23 01 2007 ozzano dell’emilia 1367 27 09 2004 san lazzaro 1511 tuber excavatum vittad. 01 11 2004 pianoro 2548 15 11 2004 san lazzaro 3190 08 11 2006 ozzano dell’emilia 3197 08 11 2006 ozzano dell’emilia 3198 08 11 2006 ozzano dell’emilia 3191 08 11 2006 ozzano dell’emilia 3189 08 11 2006 ozzano dell’emilia 3376 30 12 2006 san lazzaro 3350 23 01 2007 ozzano dell’emilia 3345 23 01 2007 ozzano dell’emilia 3185 08 11 2006 ozzano dell’emilia 3183 tuber macrosporum vittad. 08 11 2006 ozzano dell’emilia 3184 08 11 2006 ozzano dell’emilia 3186 08 11 2006 ozzano dell’emilia 3381 30 12 2006 san lazzaro 3355 23 01 2007 ozzano dell’emilia 2547 15 11 2004 san lazzaro wild boars and truffle spores 149 herbarium n. species date municipality 3194 tuber magnatum pico 08 11 2006 ozzano dell’emilia 3204 08 11 2006 ozzano dell’emilia 3193 08 11 2006 ozzano dell’emilia 3374 30 12 2006 san lazzaro 3241 15 01 2007 ozzano dell’emilia 3240 15 01 2007 ozzano dell’emilia 3239 15 01 2007 ozzano dell’emilia 3242 15 01 2007 ozzano dell’emilia 3349 23 01 2007 ozzano dell’emilia 3243 15 01 2007 ozzano dell’emilia 3255 01 01 2007 ozzano dell’emilia 2545 15 11 2004 san lazzaro 2450 tuber rufum pico 11 09 2004 pianoro 2448 11 10 2004 pianoro 2549 15 11 2004 san lazzaro 2469 27 09 2004 san lazzaro 3206 08 11 2006 ozzano dell’emilia 3200 08 11 2006 ozzano dell’emilia 3195 08 11 2006 ozzano dell’emilia 3377 30 12 2006 san lazzaro 3378 30 12 2006 san lazzaro 3347 25 01 2007 ozzano dell’emilia 1100 stephensia bombycina (vittad.) tul. 25 09 2007 san lazzaro fig. 1. spores found in wild boar faeces: a) tuber aestivum, b) tuber magnatum, c) stephensia bombycina, d) hymenogaster lycoperdineus. scale bars = 10 μm. 150 f. piattoni et al. discussion in this study a simple method to detect truffle spores in wild boars’ faeces was perfected. this method can be used for ecological studies involving the role of wild animals in hypogeous fungal spore dispersal. examining the faeces of just 14 animals hunted in two months, we were able to detect seven hypogeous fungal species including t. aestivum and t. magnatum, which are the most widespread and economically important species growing in the studied area. these results suggest that the detection of fungal spores in the faeces of wild boars may provide a first rough indication of the presence of hypogeous fungi in an table 2 wild boar data and characteristics of the spores detected in the faecal material animal code locality (municipality) sex age (months) weigth (kg) number of spores mean dimensions (μm) attempt of identification 34532 la croara (san lazzaro) ♂ 19-22 64 2 20.51 x 23.73 tuber magnatum pico 34497 via gaibola (bologna) ♂ 35 68 <10 6 12.32 x 22.58 21.52 x 22.87 hymenogaster lycoperdineus vittad. stephensia bombycina (vittad.) tul. 34526 ozzano dell’emilia ♂ 8 23 1 1 15.03 x 11.07 19.20 x 33.69 hymenogaster sp. tuber sp. 34222 montecalvo (pianoro) ♀ 7 26 1 1 19.61 x 26.32 13.38 x 29.43 tuber rufum pico hymenogaster sp. 34241 settefonti (ozzano dell’emilia) ♂ 7 26 34233 settefonti (ozzano dell’emilia) ♀ 6 27 34530 settefonti (ozzano dell’emilia) ♀ 22 65 34223 settefonti (ozzano dell’emilia) ♀ 25 69 2 4 23.78 x 27.12 13.00 x 21.54 genea verrucosa vittad. hymenogaster lycoperdineus vittad. 54321 acquafredda (pianoro) ♂ 10 32 2 21.40 x 31.03 tuber aestivum vittad. 34235 pieve (ozzano dell’emilia) ♂ 9 26 34236 sabbioni (ozzano dell’emilia) ♂ 8 26 <10 2 23.55 x 33.41 11.96 x 20.81 tuber aestivum vittad. hymenogaster lycoperdineus vittad. 54337 acquafredda (pianoro) ♀ 9 26 1 20.31 x 32.45 tuber sp. 54389 ozzano dell’emilia ♀ 7 21 54328 montecalvo (pianoro) ♀ 12 31 3 25.07 x 20.03 tuber magnatum pico wild boars and truffle spores 151 unknown area, when trained dogs are not available. this methodology may also be useful in countries were truffle harvesting is forbidden even for scientific purposes. for example, in the czech republic and in slovakia truffles are considered endangered species and they are, thus, protected by the law (grynder et al. 2011). the spores of hypogeous fungi were found in most of the animals analyzed (9/14) regardless of weight, age or sex. in the past, truffles were harvested with female pigs as it was thought that truffle scent resembled that of the male pig’s pheromone (to tuscany 2012). in fact, the steroid 5α-androst-16-en-3α-ol, which is a major component of the boar pheromone (claus et al. 1981) has been detected in the black truffle (tuber melanosporum). in the studied area, the lack of preference by female boars for truffles could be explained by the presence of truffle species different from t. melanosporum, which probably do not contain this specific pheromone. although most animals revealed the presence of hypogeous fungal spores in the faeces, the abundance of spores was poor, limited to just one or few spores in the amount of sample analyzed. this suggests that the wild boar may occasionally eat truffles and that the main damages in natural and cultivated truffières are mainly due to soil disturbance caused by excavation with the snout (moreno-arroyo et al. 2005; salerni et al. 2011; ricci 2008). on the opposite, other small mammals, like the northern flying squirrels or some marsupials, just eat truffles or simply prefer to eat truffles, whose spores accumulate in faecal pellets (lehmkuhl et al. 2004; claridge, trappe 2005). according to the claridge and trappe (2005) classification of mycophagous animals in obligate, preferential, casual, opportunistic or accidental mycophagists, the wild board can be considered an opportunistic mycophagist, ingesting truffles only occasionally, as a seasonal source of food. in mycophagous animals, there is an evidence of spore germination stimulation by the passage through the digestive system, although the effect on spore metabolic activities may differ among animals and among hypogeous fungal genera (trappe, claridge 2005). in fact, most studies on hypogeus ascoand basidiomycetes report a positive effect on spore germination (colgan, claridge 2002; claridge, trappe 2005). in contrast miller (1985) reported that the germination of spores of tuber spp. was not stimulated by digestive process in rodents. however, all the studies were carried out only on small hydnophagous mammals or marsupials and never considered large mammals like wild boars (claridge, trappe 2005). wild boars have larger movements than small sized animals, ranging between 2 and 15 km in one night and up to 300 km for males and 100 km for females during seasonal migrations (andrzejewski, jezierski 1978; singer et al. 1981; defra 2005). thus, their role in truffle long distance dispersal may be extremely important if the vitality and infectivity of truffle spores after the passage through the digestive tract, are not 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(in:) j. dighton, j.f. white, p. oudemans (eds). the fungal community. 3d.: 613–623. crc, taylor & francis, boca raton. trappe j.m., molina r., luoma d.l., cazares e., pilz d., smith j.e., castellano m.a., miller s.l., trappe m.j. 2009. diversity, ecology and conservation of truffle fungi in forests of the pacific northwest. gen. tech. rep. usda. pieri l., ventura f., vignudelli m., rossi p. 2011. nitrogen balance in a hilly semi-agricultural watershed in northers italy. italian journal of agronomy 6: 67-75. zambonelli a., rivetti c., percurdani r., ottonello s. 2000. tuberkey: a delta-based tool for the description and interactive identification of truffles. mycotaxon 74: 57–76. 2014-01-02t12:10:41+0100 polish botanical society some new records of rhizocarpon from north-eastern poland and north-western belarus vladimir v. golubkov¹ and anna matwiejuk² ¹ department of botany, faculty of biology and ecology, ya. kupala hrodna state university azheshka str. 22, by-230023 hrodna, belarus, vgolubkov@grsu.by, vgolubkov@tut.by ² department of botany, institute of biology, university of bialystok świerkowa 20b, pl-15-950 białystok, matwiej@uwb.edu.pl golubkov v. v., matwiejuk a.: some new records of rhizocarpon from north-eastern poland and north-western belarus. acta mycol. 44 (2): 201–210, 2009. new localities of rhizocarpon distinctum, r. geographicum and r. reductum are reported from ne poland. rhizocarpon distinctum, r. grande, r. hochstetteri, r. lavatum, r. petraeum, r. polycarpum and r. reductum are reported from nw belarus for the first time. rhizocarpon hochstetteri, r. lavatum and r. polycarpum are new to belarus. a key to species occurring in the area is given. key words: lichenized fungi, rhizocarpon, new localities, poland, belarus introduction rhizocarpon ramond ex dc. is a large genus of ca 200 species belonging to the family rhizocarpaceae. it is widely distributed throughout the world, especially in alpine and polar regions. species of the genus grow on rock and are long living. they have been widely used in studies on moraine dating in a procedure known as lichenometry. the species of rhizocarpon predominantly colonise siliceous rocks, although some species occur on basic substrates; a number of taxa are parasitic on other lichens (poelt, hafellner 1982; poelt, vězda 1984; holtan-hartwig, timdal 1987; poelt 1990). thomson (1967) divided the genus rhizocarpon into taxa with a yellow thallus containing rhizocarpic acid (subgenus rhizocarpon) and taxa with white, ashy or brown thalli lacking rhizocarpic acid (subgenus phaeothallus). a comprehensive taxonomic study of yellow species of rhizocarpon in europe was conducted by runemark (1956a, b), but intra-specific variation of the r. geographicum complex is still acta mycologica vol. 44 (2): 201–210 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 202 v.v. golubkov and a. matwiejuk unclear (wirth 1995). a comprehensive revision of the so-called r. obscuratum complex with taxa containing hyaline and muriform ascospores was carried by fryday (2000a) and of the so-called r. hochstetteri complex with taxa containing hyline and one-septate ascospores was carried out by fryday (2002). in the nordic countries, the taxonomy of the non-yellow species of rhizocarpon with hyaline and muriform ascospores is recognized by ihlen (2004). the knowledge of the species diversity of rhizocarpon in north-eastern poland is relatively good as reflected in literature data (e.g., zielińska 1980; fałtynowicz 1994; bystrek, kolanko 2000; kukwa, fałtynowicz 2002; cieśliński 2003; sparrius 2003; zalewska et al. 2004a, b; kolanko 2005; kubiak 2005; matwiejuk 2008), while it is insufficient in belarus. data mostly from central and eastern belarus have been reported (gorbach 1973; golubkov 1987). cieśliński (2003) reports six species from ne poland, r. distinctum, r. eupetraeum, r. geographicum, r. lecanorinum, r. obscuratum and r. polycarpum. zalewska et al. (2004b) confirmed the occurrence of five of the above species (except r. eupetraeum) and reported r. reductum from the suwalski landscape park, where the greatest concentrations of boulders in ne poland can be found. gorbach (1973) lists six species for belarus: r. geographicum, r. grande, r. distinctum, r. concentricum, r. obscuratum and r. reductum. study area the study area comprises north-eastern poland (the area acc. to cieśliński 2003) and north-western belarus. ne poland consists of two administrative areas, podlaskie and warmińsko-mazurskie voivodeships. nw belarus consists of one administrative centre, grodno. species of rhizocarpon grow on erratics and stones in the area investigated. those are postglacial boulders brought to the area by the scandinavian ice-sheet in the pleistocene. they occur individually or in larger clusters, forming boulder concentrations. granite rocks constitute approximately 80% of erratic boulders in ne poland and nw belarus, the remaining ones being gneiss rocks. the greatest boulder concentrations in ne poland can be found in the suwalski landscape park with the greatest concentration of boulders in the lowlands in the „głazowisko bachanowo” reserve comprising ca ten thousand boulders (kukwa, fałtynowicz 2002). fortifications dating from world war i in nw belarus in the vicinity of grodno are currently being investigated not only by historians but also by biologists. material and methods the material was collected during visits to respective habitats in north-eastern poland (2002–2009) and north-western belarus (1983–2008). additionally, collections of rhizocarpon deposited in the herbarium of the institute of biology, university some new records of rhizocarpon 203 of białystok and grsu, msk-l herbaria were verified. data concerning species of rhizocarpon and their distribution in ne poland was supplemented with literature data. species were verified and named according to thomson (1997), fryday (2002) and ihlen (2004). based on descriptions and recent taxonomic innovations as well as chemical, microscopic and morphological characters, an original key to all rhizocarpon species found in the study area was prepared and is presented here. chemical analyses were performed using thin-layer chromatography (tlc) according to orange et al. (2001) with the use of solvents a and c. list of species rhizocarpon distinctum th. fr., lichenogr. scand. 1: 625 (1874). syn. rhizocarpon ambiguum (naeg.) a. zahlbr.; rh. hyalescens vain. for the description of the species see ihlen (2004). spot test reactions: thallus k+ yellow, c−, kc−, pd−; medulla i+ blue, pd–, k–, c−. substances detected by tlc: stictic acid in thallus and apothecia. the species was found on acid rocks. the habitats recorded were usually open and sunny. r. distinctum is bipolar. it is known from north europe, north america, south america and antarctica. in europe it has been reported from belarus (gorbach 1973), germany (wirth 1995), the czech republic (vĕzda, liška 1999), lithuania (motiejūnaitė 1999), estonia (randlane, saag 1999), austria (hafellner, türk 2001), portugal (boom, jansen 2002), spain (llimona, hladun 2001), great britain (coppins 2002), poland (fałtynowicz 2003), fennoscandia (santesson et al. 2004; ihlen 2004), iceland (kristinsson, heidmarsson 2006). the species is known from numerous localities in north-eastern poland (zielińska 1980; cieśliński, tobolewski 1989; fałtynowicz 1994; bystrek, kolanko 2000; cieśliński 2003; zalewska et al. 2004a, b; kolanko 2005; kubiak 2005); however, it was reported from central belarus (minsk region) only by gorbach (1973) and golubkov (1987). specimens examined. ne poland. wysoczyzna białostocka high plain, białystok, cemetery, stone gravestone, oct. 2009, leg. a. matwiejuk. nw belarus. grodno region: schutchinskiy district − ca 1 km sw of orlya village, on the niemen river, granite, 6 sept. 1989, leg. v. golubkov (msk-l), vicinity of pugachi village, geological monument of nature “great stone”, 17 aug. 1999, leg. v. golubkov (msk-l), vicinity of jakubovichi village, granite, 16 june 1989, leg. v. golubkov (msk-l), vicinity of kostenevo village, granite, 10 july 1999, leg. v. golubkov (msk-l); zelvenskiy district − vicinity of savichy village (1 km sw), stones, 15 aug. 1999, leg. v. golubkov (msk-l); svislotchskiy district − ca 1 km sw of khlupin village, on ros river, granite, 11 june 1983, leg. v. golubkov (msk-l); mostovskiy district − ca 0.5 km sw of lunno village, granite, 27 apr. 1994, leg. v. golubkov (msk-l); grodno district − ca 1 km sw of kamenka village, the sixth fort of the grodno fortress, granite, 29 sept. 2007, leg. v. golubkov & e. bludov (grsu), vicinity of grodno, the second fort of the grodno fortress, granite, 11 aug. 1990, leg. v. golubkov (grsu), right riverside of the niemen, granite, 7 sept. 2004, leg. v. golubkov (grsu), 1 km nw of podłabienie village, the third fort of the grodno fortress, on moss-covered siliceous stones, 29 july 2008, leg. v. golubkov (grsu), 1.5 km nw of prigodichy village, on granite, 7 sept. 2004, leg. v. golubkov (grsu). note. it is the first record of the species for north-western belarus. 204 v.v. golubkov and a. matwiejuk rhizocarpon eupetraeum (nyl.) arnold, flora 53: 478 (1870). for the description of the species see thomson (1997). spot test reactions: thallus k+ yellow-turning red, c−, kc+ red, pd+ orange; medulla k+ yellow turning red, pd+ yellow, c–, i+ deep blue. substances detected by tlc: norstictic acid (thomson 1997). the species was found on acid rocks. the species is arctic and boreal. it is known from northern europe and north america. in europe it has been reported from germany (wirth 1995), the czech republic (vĕzda, liška 1999), lithuania (motiejūnaite 1999), austria (hafellner, türk 2001), spain (llimona, hladun 2001), poland (fałtynowicz 2003), fennoscandia (santesson et al. 2004). r. eupetraeum is known from three localities in north-eastern poland (cieśliński 2003). rhizocarpon geographicum (l.) dc., lamarck & de candolle, fl. franç., edn 3 (paris) 2: 365 (1805). for the description of the species see poelt (1988). spot test reactions: medulla i+ blue, k–, c–, pd–; uv+ bright orange. substances detected by tlc: rhizocarpic acid, psoromic acid in thallus and apothecia. the species was observed on acidic rocks, on boulders. the species is cosmopolitan in colder areas. in europe it has been reported from belarus (gorbach 1973), germany (wirth 1995), the czech republic (vĕzda, liška 1999), lithuania (motiejūnaitė 1999), estonia (randlane, saag 1999), austria (hafellner, türk 2001), portugal (boom, jansen 2002), spain (llimona, hladun 2001), great britain (coppins 2002), poland (fałtynowicz 2003), fennoscandia (santesson et al. 2004), iceland (kristinsson, heidmarsson 2006), albania (hafellner 2007). r. geographicum is known from numerous localities in north-eastern poland (see zielińska 1980; ciesliński, tobolewski 1989; fałtynowicz 1994; kukwa, fałtynowicz 2002; cieśliński 2003; zalewska et al. 2004a, b; kolanko 2005; kubiak 2005; matwiejuk 2008). it was reported from western belarus by gilibert (1781) and from central belarus (minsk region) by gorbach (1973). specimens examined. ne poland. dolina biebrzy valley, granite, oct. 2009, leg. a. matwiejuk. nw belarus. grodno region, grodno district − right riverside of the niemen, 1.5 km w of prigodichi village, granite, 7 sept. 2004, leg. v. golubkov (grsu), ca 1 km sw of kamenka village, the sixth fort of the grodno fortress, granite, 29 sept. 2007, leg. v. golubkov, o. ostrovska, d. sviridov & a. levchuk (grsu). rhizocarpon grande (flörke) arnold, flora 54: 149 (1871). syn. rhizocarpon endamyleum th. fr. for the description of the species see thomson (1997). spot test reactions: me-or the description of the species see thomson (1997). spot test reactions: medulla i+ blue, c+ red, pd–, k–. substances detected by tlc: gyrophoric acid, stictic acid and norstictic acid in thallus and apothecia. the species was found on acidic rocks. r. grande is bipolar. it is known from mongolia (golubkova 1983), north america (thomson 1997), antarctica (øvstedal, lewis smith 2001). in europe it has been reported from germany (wirth 1995), the czech republic (vĕzda, liška 1999), austria (hafellner, türk 2001), poland (fałtynowicz 2003), fennoscandia (santesson et al. 2004) and iceland (kristinsson, heidmarsson 2006). some new records of rhizocarpon 205 r. grande is known from one locality in north-eastern poland (zielińska 1980). it was reported from central belarus (minsk region) by gorbach (1973). specimens examined. nw belarus. grodno region: schutchinskiy district − vicinity of pugachi village, the “great stone” geological nature monument, 17 aug. 1999, leg. v. golubkov (msk-l); svislotchskiy district − 1 km sw of porozovo village, on the ros river, granite, 11 june1983, leg. v. golubkov (msk-l). note. it is the first record of the species from north-western belarus. rhizocarpon hochstetteri (körb.) vain., acta soc. fauna fl. fenn. 53: 280 (1922). syn. catillaria hochstetteri körber, parerga lichenol. 195 (1861); rhizocarpon applanatum (fr.) th. fr.; rh. massalongii sensu malme; rh. crenulatum h. magn. for the description of the species see fryday (2002). spot test reactions: thallus k−, c−, pd−; medulla k−, c−, pd−, i−. substances detected by tlc: lichen products not detected. the species was found on acidic rocks. r. hochstetteri is circumpolar arctic. it is known from europe, new zealand (fryday 2000b), japan (kurokawa 2003) and north america (thomson 1997). in europe it has been reported from germany (wirth 1995), austria (hafellner, türk 2001), the british isles (fryday 2002), poland (fałtynowicz 2003), fennoscandia (santesson et al. 2004). specimen examined. nw belarus. grodno region: smorgonskiy district − 1.5 km sw of balaban village, granite, 16 june 1989, leg. v. golubkov (msk-l). note. it is the first record of the species from belarus. rhizocarpon lavatum (fr.) hazsl., magyar. birod. zuzmó-flór.: 206 (1884). syn. rhizocarpon obscuratum f. lavatum (fr.) th. fr.; rh. orphninum (vain.) vain. for the description of the species see ihlen (2004). spot test reactions: thallus k–; medulla i–. substances detected by tlc: lichen products not detected. the species was found on acidic rocks. the species is known from germany (wirth 1995), the czech republic (vĕzda, liška 1999), austria (hafellner, türk 2001), spain (llimona, hladun 2001), portugal (boom, jansen 2002), great britain (coppins 2002), poland (fałtynowicz 2003), fennoscandia (santesson et al. 2004) and iceland (kristinsson, heidmarsson 2006). specimens examined. nw belarus. grodno region: smorgonskiy district − 1.5 km w of balaban village, granite, 16 june 1999, leg. v. golubkov (msk-l); grodno district − vicinity of kamenka village, granite, 27 june 1997, v. golubkov (msk-l). note. this is the first records of the species from belarus. rhizocarpon lecanorinum anders, hedwigia 64: 261 (1923). for the description of the species see poelt (1988). spot test reactions: medulla pd+ red, i+ blue, k+ yellow; epithecium k−; exciple k+ red. substances detected by tlc: psoromic acid in thallus and apothecia. the species grows on siliceous rocks, on boulders. the species is known from germany (wirth 1995), the czech republic (vĕzda, liška 1999), lithuania (motiejūnaitė 1999), estonia (randlane, saag 1999), austria (hafellner, türk 2001), spain (llimona, hladun 2001), portugal (boom, jansen 2002), great britain (coppins 2002), poland (fałtynowicz 2003), fennoscandia (santesson et al. 2004). 206 v.v. golubkov and a. matwiejuk r. lecanorinum is known from one locality in north-eastern poland (cieśliński, tobolewski 1989; see also kukwa and fałtynowicz 2002; cieślińki 2003; zalewska et al. 2004b). rhizocarpon petraeum (wulfen) a. massal., ric. lich. crost.: 102 (1852). syn. rhizocarpon concentricum auct.; rh. excentricum (ach.) arnold, verh. zool.-bot. ges. wien 29: 356 (1879); siegertia petraea (wulfen) v. wirth. for the description of the species see ihlen (2004). spot test reactions: thallus k+ yellow; medulla i–, k–, c–, pd–. substances detected by tlc: stictic acid in thallus and apothecia. the species was found on acidic rocks. r. petraeum is circumpolar arctic and boreal. in europe the species is known from germany (wirth 1995), the czech republic (vĕzda, liška 1999), lithuania (motiejūnaitė 1999), austria (hafellner, türk 2001), spain (llimona, hladun 2001), great britain (coppins 2002), iceland (kristinsson, heidmarsson 2006), poland (fałtynowicz 2003), fennoscandia (santesson et al. 2004). r. petraeum was reported from central belarus (minsk region) by gorbach (1973). specimens examined. nw belarus. grodno region: grodno district − ca 1 km sw of kamenka village, the sixth fort of grodno fortress, on moss-covered siliceous stones, 29 sept. 2007, leg. v. golubkov, o. ostrovska, d. sviridov & a. levchuk (msk-l); schutchinskiy district − vicinity of jakubowice village, granite, 16 june 1989, leg. v. golubkov (msk-l). note. this is the first record of the species from north-western belarus. rhizocarpon polycarpum (hepp) th. fr., lichenogr. scand. 1: 617 (1874). syn. rhizocarpon confervoides sensu rabenh., kremp.; rh. cyanescens (hellb.) zahlbr. for the description of the species see thomson (1997). spot test reactions: thallus k+ yellow, c–; medula i+ blue, k–, c–, pd–. substances detected by tlc: stictic acid in thallus and apothecia. this species was found on acidic rocks, on boulders. the species is bipolar. it is known from north europe, north america (thomson 1997), australia (fryday 2000b), antarctica (øvstedal, lewis smith 2001). in europe it has been reported from germany (wirth 1995), the czech republic (vĕzda, liška 1999), estonia (randlane, saag 1999), austria (hafellner, türk 2001), portugal (boom, jansen 2002), spain (llimona, hladun 2001), iceland (kristinsson, heidmarsson 2006), great britain (coppins 2002), poland (fałtynowicz 2003) and fennoscandia (santesson et al. 2004). it is known from several localities in north-eastern poland (bystrek and kolanko 2000; kukwa and fałtynowicz 2002; cieśliński 2003; zalewska et al. 2004b; kubiak 2005). specimen examined. nw belarus. grodno region: svislotchskiy district − porozovo (1 km sw), on the ros river, granite, 11 june 1983, leg. v. golubkov (msk-l). note. it is the first record of the species from belarus. rhizocarpon reductum th. fr., lichenogr. scand. 1: 633 (1874). syn. rhizocarpon obscuratum auct. not. (ach.) a. massal. for the description of the species see fryday (2000b). spot test reactions: thallus k+ yellow, c−; medulla i−, c−, k+ red, pd+ orange. substances detected by tlc: stictic acid in thallus and apothecia. the species was found on acidic rocks, on boulders. the species is circumpolar. in europe it has been reported from germany (wirth 1995), poland (fałtynowicz 2003) and fennoscandia (santesson et al. 2004; ihlen 2004). some new records of rhizocarpon 207 it is known from several localities in north-eastern poland (cieśliński, tobolewski 1989, as r. obscuratum; kukwa, fałtynowicz 2002; sparrius 2003; zalewska et al. 2004b; matwiejuk 2007). it was reported from central belarus (minsk region) by gorbach (1973). specimens examined. ne poland. wysoczyzna białostocka high plain, białystok city, cemetery, stone gravestone, aug. 2005, leg. a. matwiejuk. nw belarus. grodno region: novogrudokskiy district, 1 km sw of zhdanovitchy village, on the roc river, granite, 6 june 1989, leg. v. golubkov (msk-l). note. it is the first record of the species from north-western belarus. key to the species of rhizocarpon occurring in north-eastern poland and north-western belarus 1. thallus yellow; containing rhizocarpic acid (uv+ bright orange) ........................................................ 2 1.* thallus brown or ashy; lacking rhizocarpic acid (uv−) ......................................................................... 3 2. epithecium reddish, k+ purple-violet ............................................................................ r. geographicum 2.* epithecium brownish, k−; areoles mostly-shaped, each segment partly to entirely surrounding an apothecium .................................................................. r. lecanorinum 3. ascospores 1septate .................................................................................................................................. 4 3.* ascospores muriform .................................................................................................................................. 5 4. medulla i+ blue, k−, c−; epithecium brown, k+ violet-red ........................................ r. polycarpum 4.* medulla i−, k−, c−; epithecium green-brown, k− ........................................................ r. hochstetteri 5. ascospores hyaline or only slightly darkened with age ............................................................................ 6 5.* ascospores very soon dark ......................................................................................................................... 9 6. medulla i+ blue; epithecium red-brown, k+ purple ......................................................... r. distinctum 6.* medulla i− .................................................................................................................................................. 7 7. thallus pruinous; ascospores muriform, with 10-20 cells ..................................................... r. petraeum 7.* thallus epruinous ........................................................................................................................................ 8 8. stictic acid present, thallus k+ yellow .................................................................................... r. reductum 8.* substances not detected by tlc, thallus k– ........................................................................... r. lavatum 9. medulla k− ................................................................................................................................... r. grande 9.* medulla k+ red ..................................................................................................................... r. eupatreum discussion the lichen biota in east-central europe has been studied reasonably well. some groups of species, however, particularly rhizocarpon species, are still poorly known in many regions (e.g., belarus). as a result, many species frequent or even common in other areas of europe have not been reported from a number of countries in central-east part of the continent. twenty nine species of the genus have been reported in poland so far (fałtynowicz 2003); six species were recorded in ne 208 v.v. golubkov and a. matwiejuk poland (cieśliński 2003) and six species in belarus (gorbach 1973). of the eight taxa presented here, seven species of rhizocarpon are reported for the first time for north-western belarus, including three for belarus. the occurrence of three species reported from ne poland in literature data was confirmed in this study, with new localities recorded for them. rhizocarpon distinctum is the only common species in the study area while others are rare or very rare and were found at single localities. rocks and boulders, occurring singly or in small groups, are mostly suitable for their development in the lowland. in ne poland, the area of the suwalski landscape park covers lowland regions with a particularly large accumulation of mostly granite and gneiss boulders (kukwa, fałtynowicz 2002; zalewska et al. 2004b). boulders, stones and pebbles are in different habitat types: in the fields, balks, pastures, roads, on the slopes, in forests and their edges. among the rhizocarpon species characterized in the paper, r. geographicum and r. lecanorinum are rare in the lowlands. in nw belarus, rare species of rhizocarpon were recorded on the fortifications from world war i in grodno. acknowledgements. we wish to thank the reviewers for their valuable remarks and advice. references boom, van den p. p. g., jansen j. 2002. lichens in the upper belt of the serra da estrela (portugal). österr. zeitschr. pilzk. 11: 1–28. bystrek j., kolanko k. 2000. porosty (lichenes) puszczy knyszyńskiej. zakład poligraficzny bis, lublin, 98 pp. cieśliński s. 2003. atlas rozmieszczenia porostów (lichenes) w polsce północno-wschodniej. phytocoenosis 15 (n.s.), suppl. cartogr. geobot.15, 430 pp. cieśliński s., tobolewski z. 1989. porosty polski północno-wschodniej. i. acta mycol. 25 (1): 57–100. coppins b. j. 2002. checklist of lichens of great britain and ireland. british lichen society, huddersfield. fałtynowicz w. (ed.). 1994. porosty wigierskiego parku narodowego. parki nar. rez. przyr. 13 (3): 9–28. fałtynowicz w. 2003. the lichens lichenicolous and allied fungi of poland. an annotated checklist. w. szafer institute of botany, polish academy of sciences, kraków, 435 pp. fryday a. m. 2000a. on rhizocarpon obscuratum (ach.) massal., with notes on some related species in the british isles. lichenologist 32: 207–224. fryday a. m. 2000b. additional lichen records from new zealand 31. australasian lichenology 46: 336– 339. fryday a. m. 2002. a revision of the species of the rhizocarpon hochstetteri group occurring in the british isles. lichenologist 34: 451–477. gilibert j. e. 1781. flora lithuanica inchoata, seu enumeratio plantarum quas circa grodnam coll’egit et determinavit joannes emmanuel gilibert. j. gilibert. – grodnae: typis s.r.m. primae collectionis. s. 98, 118–119. golubkov v. v. 1987. species composition and structure of lichenoflora of the state national park belovezhskaya pushcha, 85 pages. − manuscript (deposited at viniti 22.04.87, deposition no. 2829b87). golubkova n. s. 1983. an analysis of the lichen flora of mongolia. nauka, leningrad, 248 pp. gorbach n. v. 1973. lishajniki belorussii. nauka i technika, minsk, 1–583. hafellner j. 2007. checklist and bibliography of lichenized and lichenicolous fungi so far reported from albania (version 05-2007). fritschiana 59: 1–18. hafellner j., türk r. 2001. die lichenisierten pilze österreichs-eine checkliste der bisher nachgewiesenen arten mit verbreitungsangaben. stapfia 76: 1–167. holtan-hartwig j., timdal e. 1987. notes on some parasitic rhizocarpon species. lichenologist 19: 335– 338. some new records of rhizocarpon 209 ihlen p. g. 2004. taxonomy of the non-yellow species of rhizocarpon (rhizocarpaceae, lichenized ascomycota) in the nordic countries, with hyaline and muriform ascospores. mycological research 108: 533–570. kolanko k. 2005. porosty biebrzańskiego parku narodowego i terenów przylegających. (in:) a. dyrcz, c. werpachowski (eds). przyroda biebrzańskiego parku narodowego. monografia. biebrzański park narodowy. osowiec twierdza, 149–160. kristinsson h., heidmarsson s. 2006. checklist of lichens in iceland. internet version. http:��www.fl orais-checklist of lichens in iceland. internet version. http:��www.floraislands.is�fletlist.htm kubiak d. 2005. lichens and lichenicolous fungi of olsztyn town (ne) poland. acta mycol. 40 (2): 293– 332. kukwa m., fałtynowicz w. 2002. porosty rezerwatu „głazowisko bachanowo nad czarną hańczą” i przyległego lasu łęgowego w suwalskim parku krajobrazowym. parki nar. rez. przyr. 21 (4): 375–84. kurokawa s. (ed.) 2003. checklist of japanese lichens. national science museum, tokyo, 128 pp. llimona x., hladun n. l. 2001. checklist of the lichens and lichenicolous fungi of the iberian peninsula and balearic islands. bocconea 14: 1-581. matwiejuk a. 2008. nowe stanowisko rhizocarpon geographicum (rhizocarpaceae) w polsce północnowschodniej. fragm. florist. geobot. 15 (2): 25–27. motiejūnaitė j. 1999. checklist of lichens and allied fungi of lithuania. botanica lithuanica 5 (3): 251– 269. orange a., james p. w., white, f. j. 2001. microchemical methods for the identification of lichens. british lichen society, london, 101 pp. øvstedal d. o., lewis smith, r. i. 2001. lichens of antarctica and south georgia. a guide to their identification and ecology. cambridge, cambridge university press, 411 pp. poelt j. 1990. parasitische arten der flechtengattung rhizocarpon: eine weiter übersicht. mitteilungen der botanischen staatssamlung münchen 29: 515–538. poelt j., hafellner j. 1982. rhizocarpon vorax spec. nov. (lichenes) und seine beutegenossen auf pertusaria. herzogia 6: 309–321. poelt j., vězda a. 1984. rhizocarpon inimicum spec. nov. eine weitere parasitische flechte auf lecanora rupicola spec. coll. herzogia 6: 469–475. randlane t., saag a. 1999. second checklist of lichenized, lichenicolous and allied fungi of estonia. folia cryptog. estonica 34: 1–132. runemark h. 1956a. studies in rhizocarpon i. taxonomy of the yellow species in europe. opera botanica 2 (1): 1–152. runemark h. 1956b. studies in rhizocarpon ii. distribution and ecology of the yellow species in europe. opera botanica 2 (2): 1–150. santesson r., moberg r., nordin a., tønsberg t., vitikainen o. 2004. lichen-forming and lichenicolous fungi of fennoscandia. uppsala, museum of evolution, uppsala university, 359 pp. sparrius l. b. 2003. contribution to the lichen floras of the białowieża forest and the biebrza valley (eastern poland). herzogia 16: 155–160. thomson j. w. 1967. notes on rhizocarpon in the arctic. nova hedwigia 14: 421–481. thomson j. w. 1997. american arctic lichens 2. the microlichens. madison, university of wisconsin press, 675 pp. vězda a., liška j. 1999. katalog lišejníků české republiky. institute of botany, academy of sciences of the czech republic, průhonice. wirth v. 1995. die flechten baden-württembergs. bd.1-2, ii aufl. stuttgart, verl. eugen ulmer. zalewska a., fałtynowicz w., krzysztofiak a., krzysztofiak l., picińska-fałtynowicz, j. 2004a. lichens of romincka primeval forest. (in:) a. zalewska, w. fałtynowicz (eds). lichens of the protected areas in the eurogion niemen. “man and nature” association, suwałki, 51–110. zalewska a., fałtynowicz w., krzysztofiak a., krzysztofiak l., picińska-fałtynowicz j. 2004b. lichens of suwalski landscape park. (in:) a. zalewska, w. fałtynowicz (eds). lichens of the protected areas in the eurogion niemen. “man and nature” association, suwałki, 5–50. zielińska j. 1980. porosty głazów narzutowych wysoczyzn podlaskich. acta mycol. 5: 135–148. 210 v.v. golubkov and a. matwiejuk nowe notowania gatunków rhizocarpon z polski północno-wschodniej i północno-zachodniej białorusi streszczenie celem niniejszej pracy jest uzupełnienie danych o porostach z rodzaju rhizocarpon na terenie polski północno-wschodniej i północno-zachodniej białorusi. na terenie ne polski odnotowano trzy nowe stanowiska rhizocarpon − r. distinctum, r. geographicum i r. reductum, a na terenie nw białorusi osiem gatunków − r. distinctum, r. geographicum, r. grande, r. hochstetteri, r. lavatum, r. petraeum, r. polycarpum, r. reductum. siedem gatunków (r. distinctum, r. grande, r. hochstetteri, r. lavatum, r. petraeum, r. polycarpum, r. reductum) jest podanych po raz pierwszy dla zachodniej białorusi, a trzy gatunki (r. hochstetteri, r. lavatum, r. polycarpum) są nowe dla białorusi. opracowano autorski klucz do gatunków występujących na badanym obszarze. 2014-01-01t11:49:28+0100 polish botanical society zasmidium cellare in poland andrzej chlebicki and magdalena majewska department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, a.chlebicki@botany.pl chlebicki a., majewska m.: zasmidium cellare in poland. acta mycol. 45 (1): 121–124, 2010. morphology and polish localities of zasmidium cellare (pers.) fr. are presented. the appropriate name of the fungus is discussed. key words: cellar mould, distribution, taxonomy introduction fungi can be found in various environments, among them in wine, beer and brandy storage cellars and warehouses. there are often found ubiquitous microfungi from such genera as alternaria nees, aspergillus p. micheli ex haller, cladosporium link, penicillium link, rhizopus ehrenb., verticillium nees, ulocladium preuss as well as aureobasidium pullulans (de bary) arnaud s. lato, baudoinia compniacensis (richon) j. a. scott & unter. and zasmidium cellare (pers.) fr. (goto et al. 1989; scott et al. 2007). zasmidium cellare can utilize various alcohols and esters, acetic acid, acetylaldehyde, formaldehyde and thymol as sole carbon sources. fungal cultures exposed to vapours of these carbon sources exhibited abundant growth (schanderl 1958). zasmidium cellare is typical wine cellar mould. hawksworth and riedl (1977) restricted the name zasmidium cellare to the mycelial state only and proposed a new name, rhinocladiella ellisii hawksworth, for the conidial state. de hoog (1979) pointed out that z. cellare infrequently produces conidia. in spite of the lack of the description of conidial state in the original publication (persoon 1794), structures suggestive conidia were found in authentic specimens from l herbarium (de hoog 1979). morphologically zasmidium resembles stenella syd., however the type species of stenella (s. arguata syd.) clusters phylogenetically with members of the teratosphaeriaceae (arzanlou et al. 2007). the nucits sequences of z. cellare revealed closest relationships with members of the dothideomycetidae (mycosphaerellales, mycosphaerellaceae) rather than the chaetotyriales (de hoog et al. 1999; wilkinson et al. 2005; belliveau, bärlocher 2005). acta mycologica vol. 45 (1): 121–124 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 122 a. chlebicki and m. majewska zasmidium cellare (pers.) fr. summa veg. scand. 2: 407, 1849. bas.: racodium cellare pers. syn.: cladosporium cellare (pers.) schanderl, rhinocladiella cellaris (pers.) m. b. ellis, rhinocladiella ellisii hawskworth hyphae up to 5000 μm in length, 2-2.2 μm wide, wall 0.5-0.6 μm thick, aerial hyphae verrucose, rarely encrusted, pale brown to olivaceous brown, branched, with thin septa (fig. 1), conidia not observed. zasmidium cellare was the first time mentioned from poland by schneider (1875), then by schroeter (1883) and smyk (1954). however no localities were given by two first authors. it is interesting that j. schroeter composed a humourous song about the cellar mould (see polish translation at the end of this article) in which zasmidium cellare was attributed to lower silesia, in particular to the hungarian famous town – of tokaj, where the fungus grows luxuriantly. smyk (1954) noted z. cellare in malts from dobrodzień (opole voivodeship), facimiech (małopolska voivodeship) and gwoździany (śląsk voivodeship) in poland, although no mention was made of morphological details of the fungus. świderska-burek (2008) following the opinion of dugan et al. (2004), wrongly treated zasmidium cellare as a synonym of rhinocladiella ellisii hawksworth because of the absence in the polish literature of data of its fig. 1. zasmidium cellare in the brandy maturation cellar of the ‘polmos’ manufacture in szcze-szczecin, poland: a – colonies on rock wall; b – verrucose hyphae; c – young hyphae; d – encrusted hyphae. scale bar for b, c, d = 20 μm. zasmidium cellare in poland 123 morphological characterization. properly this latter taxon is applied to the conidial state of z. cellare. locality. poland, szczecin town, brandy maturation cellar of “polmos” s.a., jagiellońska str. 63/64, 26 november 2009, coll.: a. chlebicki, kram f-48618. we examined a cellar colonized by z. cellare in szczecin (fig. 1). several other fungal species were found growing in association with zasmidium cellare on the cellar walls including: penicillium chrysogenum thom, aureobasidium pullulans var. melanogenum herm.-nijh., aspergillus candidus link, papulospora preuss, sporobolomyces kluyver & c. b. niel and cladosporium herbarum (pers.) link. the species can grow on various media including maa, pda and cornmeal agar (tribe et al. 2006) as well as media containing low-levels of carbohydrate (e.g., 10 g/l glucose + 10 g/l peptone) (scott in lett.). acknowledgements. we thank dr. j. a. scott for valuable comments and correction of english. the study was supported by the ministry of science and higher education, poland (project no. n n304 328336). references arzanlou m. groenewald j. z., gams w., braun u., shin h.-d. and crous p.w. 2007. phylogenetic and morphotaxonomic revision of ramichloridium and allied genera. stud. mycol. 58: 57–93. belliveau m. j. r., bärlocher f. 2005. molecular evidence confirms multiple origins of aquatic hyphomycetes. mycol. res. 109: 1407–1417. dugan f. m., schubert k., braun u. 2004. check-list of cladosporium names. schlechtendahlia 11: 1–103. goto s., takayama k., shinohara t. 1989. occurrence of molds in wine storage cellars. journal of fermentation and bioengineering 68 (4): 230–232. hawksworth d., riedl h. 1977. nomina conservanda proposita 427. taxon 28: 347–348. hoog g. s. 1979. nomenclatural notes on some black yeast-like hyphomycetes. taxon 28 (4): 347–348. hoog g. s., zalar p., urzi c., de leo f., yurlova n. a., sterflinger k. 1999. relationships of dothideaceous black yeasts and meristematic fungi based on 5.8s and its2 rdna sequence comparison. stud. mycol. 43: 31–37. persoon c. h. 1794. neuer versuch einer systematischen einteilung der schwämme. 76. racodium, römer’s neues magazin der botanik 1: 123. schanderl h. 1958. ein zwanzigjähriger ernährungsversuch von cladosporium-arten, insbesondere cladosporium cellare mit flüchtigen organischen verbindungen. zentralblatt für bakteriologie und parasitenkunde abt ii, 111: 116–120. schneider w. g. 1875. eine anzahl für das herbarium der gesellschaft bestimmte schlesische pilze. jahres-bericht der schlesischen gesellschaft für vaterländische cultur 52: 90–91. schroeter j. 1883. zwolfte wander-versammlung und ausserordentlichen sitzung der botanischen section am sommtag, den 18 juni 1882. bemerkungen über kellerund grubenpilze. i. jahres-bericht der schlesischen gesellschaft für vaterländische cultur 61: 208–209. scott j. a., untereiner w. a., ewaze o. j., doyle d. 2007. baudoinia, a new genus to accommodate torula compniacensis. mycologia 99 (4): 592–601. świderska-burek u. 2008. hyphomycetes from ramulispora to stemphylium (in:) w. mułenko, t. majewski, m. ruszkiewicz-michalska (eds). a preliminary checklist of micromycetes in poland. (in:) z. mirek (ed.). biodiversity of poland 9. w. szafer institute of botany, polish academy of sciences, kraków: 481–494. smyk b. 1954. studia nad mikroflorą słodów. studies of the microflora of malts. roczniki nauk rolni-rolniczych ser. a, 69: 409–470. tribe h. t., thines e., weber r. w. s. 2006. moulds that should be better known: the wine cellar mould racodium cellare persoon. mycologist 20: 171–175. 124 a. chlebicki and m. majewska zasmidium cellare w polsce streszczenie w artykule omówione są problemy taksonomiczne wynikające ze specyficznej morfologii grzyba oraz podano rozmieszczenie tego gatunku w polsce. pomimo, że pierwsze wzmianki o tym gatunku pochodzą już z xix wieku, nie jest znany żaden opis morfologii polskich szczepów z. cellare. dołączono oryginał i polskie tłumaczenie piosenki napisanej na cześć z. cellare dedykowanej dr beinertowi przez johana schroetera z wrocławia (schroeter 1883): siedząc sobie w przytulnym pokoiku otoczony zapachem winorośli wezwałem właściciela który miał mi podać najlepsze ze swych win. aż nagle zobaczyłem – cóż za trwoga! korek okłada czarna pleśń gruba niczym filcu włosie rhacodium cellare! in trautem stübchen sass ich hier, umhüllt vom duft der reben; ich rief den wirth, der musste mir vom allerbesten geben; da sah ich plötzlich, welch ein schreck! den kork umhüllt ein schwarzer fleck, ein filz von dickem haare: rhacodium cellare! właściciel roześmiał się delikatnie i zrzucił z filcu zatyczkę z najjaśniejszych pereł węgierskich win skrzą się w szklanicy krople złota ‘ujrzałeś więc panie czym jest grzyb, wygląda na to jedyny, prawdziwy rhacodium cellare! da lacht der wirth ganz leis und fein und zieht den filz vom stopfen; da perlt der hellste ungarwein ins glas in goldnen tropfen. “da sieht man den gelehrten herrn: das ist ein pilz, den sieht man gern; es ist das einzig wahre rhacodium cellare! jest to grzyb ducha i mocy co szczypta ta określa oznacza dla mnie najprzedniejsze na węgrzech, najwspanialsze z win. nawet najgorszy klient wyczuwa iż ten „mech” co na szczycie ujrzał jest to niezwykle rzadkie rhacodium cellare! “das ist ein pilz von geist und kraft, der schluckt mir stets das beste; an ungarns feinsten traubensaft kneipt er allein sich feste; und auch der schlimmste kunde traut, wenn er das moos am kopf erschaut, drum heg’ ich stets das rare rhacodium cellare” ponieważ jest grzybem tak dobrym co siedzi przy butelczynach w piwnicy a to prostym nie jest wcale.’ jak gorąca moja krew w głowie coraz bardziej się rozjaśnia choć nogom trudniej jest i portfel już pustką zieje lecz rośnie bębnieniem donośnym rhacodium cellare! seitdem bin ich dem pilz so gut und sitz bei ihm im keller; da wird so leicht, so warm mein blut, im kopf wird’s immer heller; die beine freilich werden schwer, und auch das portemonnaie wird leer, doch wuchert drum das baare rhacodium cellare! jedynym jest grzybowym „mchem” lekko podniesionym ci co uwierzyli zwolennikami się stali. myślałem ze zdumieniem że oto żyje tu już lat pięćdziesiąt w ten sam sposób wspaniałe rhacodium cellare! einst ward ein pilzummoostes haupt behutsam aufgezogen; da kam, wer hätte das geglaubt, eine flieg’ herausgeflogen; ich dacht erstaunt in meinem sinn: die lebt schier fünfzig jahr da drin! so macht’s das wunderbare rhacodium cellare! kiedyś będę martwy będziecie grzebać mnie z beczkami pełnymi węgierskiego wina. będę mógł się odświeżyć pić na mej stypie sto butelek węgrzyna a owijać się będzie dokoła mego katafalku rhacodium cellare! werd ich dereinst gestorben sein, so sollt ihr mich begraben mit einem fass voll ungarwein, dass ich mich dran kann laben; dann trinkt zu meinem leichenschmaus noch hundert flaschen ungar aus, und schlingt um meine bahre rhacodium cellare! dziś wszyscy chcemy z przyjaciółmi póki żyjemy na wesoło upić się w pestkę. kielichy niech dźwięczą możliwie często ta zabawa od nowa szczęściem wesołością niech nas cieszy przez długie, długie lata rhacodium cellare! doch heute wollen wir allein der hellen freude leben, auf fröhliches zusamensein die gläser klingend heben. mög sich noch oft dies fest erneun, mit glück und frohsinn uns erfreun durch lange lange jahre rhacodium cellare! 2014-01-01t11:50:46+0100 polish botanical society lichens of abandoned zinc-lead mines urszula bielczyk1, monika jędrzejczyk-korycińska2 and józef kiszka1 1institute of biology, pedagogical university of kraków, podbrzezie 3 pl-31-054 kraków, bielczyk@up.krakow.pl 2department of plant systematics university of silesia, jagiellońska 28, pl-40-032 katowice, mjedrzej@us.edu.pl bielczyk u., jędrzejczyk-korycińska m., kiszka j. : lichens of abandoned zinc-lead mines. acta mycol. 44 (2): 139–149, 2009. a list of lichens from areas of zinc-lead ores in southern poland and a review of the characteristic lichen biota of these sites is provided. in spite of the devastated and heavy metal contaminated environment, a highly diverse epigeic and epilithic lichen biota was found, including species characteristic of various anthropogenic habitats, particularly zinc and lead enriched substrates (diploschistes muscorum, steinia geophana, sarcosagium campestre, vezdaea aestivalis and v. leprosa). also, the high-mountain species leucocarpia biatorella, as well as very rare in europe thelocarpon imperceptum, and several species categorized as very rare, endangered and protected in poland were recorded. crustose lichens are the most abundant; among fruticose forms cladonia spp. predominate and stereocaulon incrustatum is common. key words: lichenized fungi, anthropogenic habitats, heavy metals, galena, silesian-kraków upland introduction the largest resources of zinc and lead ores in poland occur in silesian-krakow monocline, where they have been mined and processed since the middle-ages. those deposits are often bound to the triassic carbonate rocks – ore-bearing dolomites (żabiński 1960). the oxidized zinc and lead ores (galena) are often defined as galmani (żabiński 1978). due to the mining and metallurgy in southern poland, numerous scars to the landscape have been developed such as excavations (voids), shafts, mining adits and mine waste heaps (molenda 1963). such activities have significantly influenced the status of surface and ground water, soils, and consequently the flora and fauna; in addition, the presence of heavy metals (zinc, lead, cadmium) considerably influenced organisms, such species as colonize such habitats have various acta mycologica vol. 44 (2): 139–149 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 140 u. bielczyk et al. features which help them to adapt to these adverse, or even toxic, conditions. zinc and lead ores are colonized those organisms with a wide tolerance of environmental stress, producing unique communities with an interesting biology (jędrzejczyk 2004; szarek-łukaszewska, grodzińska 2008). in habitats contaminated by heavy metals, lichens, usually accompanied by mosses, are of great importance, forming unique plant communities (wirth 1972; purvis, halls 1996; paus 1997; cuny et al. 2004). thus, mining or metallurgy sites provide a particular field laboratory to follow natural processes of interest to lichenologists in terms of their taxonomy, physiology, threats and protection, monitoring abilities, etc. in europe, 291 lichen species have been noted from habitats rich in metals such as iron, copper, zinc, lead, chromium, nickel (purvis, halls 1996; heibel 1999; cuny et al. 2004). lichens often thrive on substrates rich in zinc and lead, and some genera such as gyalideopsis, sarcosagium, steinia and vezdaea appear to be limited to such habitats and can be used as good indicators of zn and pb (cuny et al. 2004). some new species have been described from the zn and cd polluted areas, such as micarea confusa (coppins, van den boom 1995), pyrenocollema chlorococcum (aptroot, van den boom 1998) and coppinsia minutissima (lumbsch, heibel 1998). limited information on species composition and their behaviour on post-exploitation areas (e.g., seaward, bylińska 1980; kiszka 2003), but there has been no detailed documentation of lichen species from zinc and lead ores areas in poland. the results presented below sum up current knowledge and establish a basis for future studies on the characteristic lichen biota of abandoned zinc-lead mining areas in poland, as well contributing to our knowledge of species distribution. study area studies were carried out in five sites of former mining and metallurgical sites based on zinc and lead ores, where there has been no subsequent development and spontaneous succession of vegetation has occurred (fig.1). studies were conducted on the three mineral deposit zones of the silesian-kraków monocline: tarnowskie górybytom area (two sites in tarnowskie góry), chrzanów-jaworzno area (grassland in jaworzno-długoszyn and in balin near chrzanów), and siewierz-olkusz area (in bolesław). study sites vary in age and nature of their development. bolesław (fig.1, i). soil-rock bank and grassland (4.5 km2) covered partially by shrubs and trees (coniferous and sporadically deciduous). on reasonably large area was planted with pine during restoration work. in the s-e part, an area is protected as “ecological arable land” [in polish: użytek ekologiczny] due to the occurrence of biscutella laevigata. “warpie” wood in balin near chrzanów (fig.1, ii). this study area (1.8 km2) is covered by pine forest, with some birch and larch. forest bottom morphology suggest a post-mining character of the area. part of the study site is open grassland, with blackthorn shrubs. some arable land is present in the vicinity. jaworzno długoszyn (fig.1, iii). this study area (3.5 km2) is situated is within the city limits of jaworzno, where signs of zinc-lead ore mining are still visible between 142 u. bielczyk et al. material and methods data gathered in 2002-2004 were analyzed, the collected species being assigned to appropriate morphological and ecological groups. collected material was analysed according to standard morphological and anatomical methodology. in this paper we follow fałtynowicz (2003) lichen nomenclature, excluding the genus coenogonium (kauff, büdel 2005), and mirek et al. (2002) for plant nomenclature. documentation and collected material is deposed in the herbarium of the department of biology, institute of biology, pedagogical university of kraków (krap-l). for the identified species, their frequency on the study sites was determined according to three categories: 1 – rare (1-3 records), 2 – frequent (4-10 records) and 3 – common (over 10 records) on one observed sites. results the number of known lichen species from the post-mining, ore-bearing areas of the silesian-kraków upland reached 89 taxa, of which 76 are listed in table 1. table 1 lichen species in studied sites b – bolesław, w – “warpia” wood in balin, jd – jaworzno-długoszyn, tgd – tarnowskie góry dolomite spoil heap, tgp – tarnowskie góry “planeta” wood no taxon substrate preferency/ies frequency scale b w jd tgd tgp 1. agonimia tristicula (nyl.) zahlbr. soil, bryophytes 1 2. arthonia exilis (flörke) anzi thymus sp. 1 3. arthonia lapidicola (tayl.) branth & rostr. pebbles 1 4. aspicilia contorta (hoffm.) kremp. subsp. hoffmanniana ekman & fröberg stones 1 5. aspicilia moenium (vain.) g. thor & timdal stones 1 6. bacidia bagliettoana (a. massal. & de not.) jatta soil, bryophytes 3 3 3 7. bacidina phacodes (körb.) vězda soil, bryophytes 2 2 2 8. baeomyces rufus (huds.) rebent. soil 3 2 9. caloplaca holocarpa (hoffm.) a. e. wade stones, pebbles 1 10. candelariella aurella (hoffm.) zahlbr. stones, pebbles 2 2 3 2 11. cetraria islandica (l.) ach. soil 2 2 2 12. cladonia cariosa (ach.) spreng. soil 1 13. cladonia cervicornis subsp. verticillata (hoffm.) ahti soil 2 14. cladonia coniocraea auct. bark 1 15. cladonia fimbriata (l.) fr. soil 1 16. cladonia furcata (huds.) schrad. soil 3 3 2 17. cladonia glauca flörke soil 2 2 3 18. cladonia pocillum (ach.) grognot soil, bryophytes 3 3 3 19. cladonia pyxidata (l.) hoffm. soil, bryophytes 3 2 3 3 2 20. cladonia rangiformis hoffm. soil 1 2 21. cladonia subulata (l.) weber soil 2 2 lichens of zinc-lead mines 143 22. cladonia symphycarpia (flörke) fr. soil 2 2 2 2 23. coeniogonium pineti (ach.) lücking & lumbsch bark 1 3 24. collema limosum (ach.) ach. soil 2 25. collema tenax (sw.) ach. soil 1 1 2 26. diploschistes muscorum (scop.) r. sant. cladonia spp., bryophytes 3 3 2 27. diploschistes scruposus (schreb.) norm. stones 1 2 28. endocarpon pusillum hedw. soil 1 29. hypocenomyce scalaris (ach. ) m. choisy bark 1 2 30. hypogymnia physodes (l.) nyl. bark 1 2 31. lecania cyrtella (ach.) th. fr. dianthus sp. 1 32. lecanora albescens (hoffm.) branth & rostr. rock, stones 2 2 33. lecanora conizaeoides cromb. bark 2 3 3 3 3 34. lecanora dispersa (pers.) sommerf. rock, stones 3 3 2 35. lecanora hagenii (ach.) ach. wood 2 36. lecanora saligna var. sarcopis (ach.) hillmann bark 2 2 2 37. lecidella stigmatea (ach.) hertel & leuckert rock 1 38. lepraria sp. bark 2 2 39. leptogium biatorinum (nyl.) leight. soil 2 40. micarea denigrata (fr.) hedl. wood 1 2 41. micarea prasina fr. bark 2 3 3 42. mycobilimbia tetramera (de not.) vitik., ahti, kuusinen, lommi & t. ulvinen soil, bryophytes 3 3 43. myxobilimbia sabuletorum (schreb.) hafellner soil, bryophytes 1 44. parmelia sulcata taylor bark 1 45. peltigera didactyla (with.) j.r. laundon soil, bryophytes 2 2 46. peltigera rufescens (weiss) humb. soil 2 3 2 47. phaeophyscia nigricans (flörke) moberg stones 1 48. phaeophyscia orbicularis (neck.) moberg stones 2 49. physcia adscendens (fr.) h. olivier stones 2 50. physcia caesia (hoffm.) fürnr. rock, stones 2 51. physcia tenella (scop.) dc. bark 1 52. placynthiella icmalea (ach.) coppins & p. james bark 1 2 2 53. polyblastia albida arnold rock 1 54. porpidia crustulata (ach.) hertel & knoph stones 1 2 55. protoblastenia rupestris (scop.) j.steiner rock, 1 56. protoparmeliopis muralis (schreb.) choisy rock, stones 1 57. sarcosagium campestre (fr.) poetsch & schied. soil, bryophytes 3 2 58. scoliciosporum chlorococcum (stenh.) vězda bark 3 3 3 3 3 59. scoliciosporum umbrinum (ach.) arnold stones 2 60. steinia geophana (nyl.) stein soil, bryophytes 3 61. stereocaulon incrustatum flörke soil 2 62. strangospora moriformis (ach.) stein wood 1 63. thelidium papulare (fr.) arnold stones 1 64. trapelia coarcatata (sm.) m. choisy in werner stones 1 2 2 65. trapelia placodioides coppins & p. james stones 1 66. trapeliopsis flexuosa (fr.) coppins & p. james soil, bryophytes 2 2 67. verrucaria aethiobola wahlenb. rock, stones 2 2 2 2 68. verrucaria bryoctona (th. fr.) orange soil, bryophytes 2 2 2 69. verrucaria dolosa hepp rock, stones 2 2 2 70. verrucaria fatrana servít stones 2 71. verrucaria muralis ach. stones, pebbles 3 2 3 3 2 72. verrucaria nigrescens pers. stones, pebbles 3 2 2 73. verrucaria obfuscans nyl. stones 2 74. verrucaria procopii servít stones, pebbles 2 2 75. vezdaea aestivalis (ohlert) tscherm.-woess & poelt soil, bryophytes 1 2 1 76. vezdaea leprosa (p. james) vězda soil, bryophytes 1 2 2 2 144 u. bielczyk et al. a clearer picture of lichens is completed by 13 other species published earlier: leucocarpia biatorella (kiszka, kościelniak 2006), thelocarpon imperceptum (kiszka 2006), cetraria aculeata, cladonia foliacea, sarcogyne regularis and thelidium velutinum (kiszka, szarek-łukaszewska 2006), micarea botryoides, m. micrococca (czarnota 2007), cladonia squamosa, c. arbuscula subsp. mitis, lepraria elobata, l. jackii and candelariella reflexa (pawlik-skowrońska et al. 2008). studied area, in spite of its serious environmental transformation, can be characterized by the lichen species richness, but the number of species on a particular site varies according to surface, size, local edaphic conditions, degree of transformation. only four species were noted from all five study sites, namely cladonia pyxidata, lecanora conizaeoides, scoliciosporum chlorococcum and verrucaria muralis. candelariella aurella, cladonia symphycarpia, vezdaea leprosa and verrucaria aethiobola were noted from four sites, 18 species from 3 sites, and 33 from only one. the highest diversity, 69 species, was found in bolesław, of which 25 were only found at that site. species abundance also varies, and most of them are rare and very rare species. they form the most interesting part of the lichen biota and give the studied area distinctive and characteristic features. taxonomically the most numerous groups are represented by cladonia and verrrucaria, with 13 and 8 species respectively. morphologically, crustose lichens are dominant (62 species), many of which have small, often inconspicuous thalli; some are pioneer species, with an ability of to rapidly colonize rocks and soils. foliose forms are represented by 17 species, and only 10 species have fruticose forms. every type of substrate available for lichens was investigated. terricolous lichens (40 species), growing directly on soil or on mosses, or plant debris, were dominant. diploschistes muscorum is particularly interesting due to its specific biology: in the first phase of the development it grows on other lichen thalli (especially on cladonia species), but afterwards colonizes plant debris, soil or gravel. epilithic lichens (29 species) are found on rock exposures, gravel, small stones and local ore, as well as imported building materials. only small fraction of recorded species were epiphytic due to the shortage of forest complexes or old trees. at the same time, epiphytic lichens are most vulnerable to atmospheric pollution, so were represented by common, toxitolerant species such as scoliciosporum chlorococcum and lecanora conizaeoides. hypogymnia physodes rarely occurs and parmelia sulcata was noted only once. due to the lack of suitable habitats, epixylic lichens are also scarce, but on small pieces of wood lying on the ground, lecanora hagenii, micarea denigrata and strangospora moriformis were noted. discussion the number of species noted from zinc-lead ore-bearing areas in poland is relatively high and can be compared to analogous lists from other european areas. purvis and halls (1996) mention 61 species of epigeic lichens characteristic of lead/zincrich environments from the united kingdom, belgium, the netherlands, france and germany. the species composition of lichen on metalliferous sites is very diverse lichens of zinc-lead mines 145 and depends not only on the heavy metals present, but also on other edaphic characteristics, such as the chemical and physical features of the substrate (cuny et al. 2004) and local microclimatic conditions. the biodiversity depends on the ore type and its chemical composition, post-mining waste granularity, storage method, land restoration procedures, and level of air pollution. the current status of the environment in the vicinity of mines and heavy metal processing works does not favour epiphytic lichens, especially the toxic effect of sulfur dioxide (kiszka 1993) which is exacerbated by high concentrations of heavy metals in the air (godzik 1993). studies carried on selected epiphytic lichen species from bukowno showed high accumulations of zn and pb in their thalli (pawlikskowrońska et al. 2008). other features can be observed in the case of terricolous and epilithic species which rapidly and effectively colonize such substrates. some, for example protoparmeliopsis muralis, are common in various anthropogenic habitats. others, of similar characteristics, but rarely noted due to their inconspicuous size and short existence, such as steinia geophana and sarcosagium campestre, are ruderals (gilbert 1990). vezdaea leprosa and v. aestivalis, recently found on many sites in poland (czarnota, kiszka 2004), were frequently noted, and other species, regularly observed in europe on zn and pb substrates, included bacidia bagliettoana, cladonia cariosa, c. glauca, c. furcata, c. rangiformis, collema tenax, diploschistes muscorum, myxobilimbia sabuletorum, scoliciosporum umbrinum and trapelia coarctata. yet, it would be hard to classify which of them are obligatory metallophytes and which are metallotolerant. it is though doubtless that diploschistes muscorum is simultaneously tolerant and indicator for zn and pb presence in the substrate, species very frequent on the studied area. in the case of fruticose species, cetraria islandica, c. aculeata and many cladonia species, as well as several stereocaulon species, especially s. nanodes, are characteristic of zinc and lead habitats. in poland, only stereocaulon incrustatum was noted, but not recorded from the similar areas; in the studied area it is quite common and forms a large population, with high concentration of zn and pb in their thalli (pawlik-skowrońska et al. 2008). in spite of the high level of environment degradation and pollution, the postmining and zinc-lead ore processing areas, support a number of unique and exceptional lichens. this specific feature of the ore-bearing part of the silesian-kraków upland is manifested by the presence of several interesting plant species, with the extremely rare biscutella laevigata, recorded only from tatra mountains (wierzbicka, pielichowska 2004) or arabidopsis halleri, which accumulates extremely high concentrations of various heavy metals (pauwels et al. 2006). from a lichenological point of view, apart from lichen species mentioned above, mention should be made of other rarities in poland, namely leptogium biatorinum, arthonia exilis, endocarpon pusillum, collema limosum, polyblastia albida, thelidium papulare and verrucaria fatrana. species protected by law, such as cetraria aculeata, c. islandica, peltigera didactyla, p. rufescens and stereocaulon incrustatum were also recorded, and seven species listed are endangered, with en, vu and nt threat categories (cieśliński et al. 2006). two species, peltigera rufescens and p. didactyla have especially interesting ecology, as they contain symbiotic cyanobacteria in their thalli, with the ability to fix atmospheric nitrogen, and as such are an important part of trophic chain. apart from the species defined as toxitolerant, genuine lichen rarities, such as peltigera venosa, normally an arctic-alpine species, can be found on metalliferous 146 u. bielczyk et al. sites in lowland areas (purvis, halls 1996); montane species were also found in postmining, lowland areas in poland. leucocarpia biatorella (kiszka, kościelniak 2006), to date known only from high elevations in the tatras and western beskidy mts. (olech 1999; flakus 2007), was also recorded. another species, agonimia tristicula, found in the study area, although widely distributed in some parts of europe, is rare in poland where to date it has only been recorded from the carpathians (olech, kiszka 1999; kościelniak 2004; flakus 2007). the extremely rare terricolous thelocarpon imperceptum was also found on a restored post-mining site in the vicinity of bolesław (kiszka 2006); it is exceptionally rare in europe, to date being recorded only from switzerland (locus classicus) and russia (salisbury 1966), and recently from the netherlands (van den boom 2000). conclusions the studies herewith provide a basis for the statement that the post-mining areas of the silesian-kraków upland is a unique region for lichen species, not only for poland, but also at the european scale. these studies are on-going within the project „vegetation of calamine soils and its importance for biodiversity and landscape conservation in post-mining areas” (fm eea pl 0265). this work will be crucial for a better understanding of ecosystem functioning under stress conditions in areas subjected to post-mining and post-processing zinc-lead ores. at the same time, the complex character of these sites, including various edaphic factors, will increase our knowledge of the ecological conditions and the tolerance of heavy metal concentrations by various species, and the value of particular lichen species as bioindicators. a further aim of these studies is to find suitable methods for biodiversity restoration of degraded areas. effective restoration may in turn provide an opportunity for the continued existence of many interesting lichen species. polish zinc-lead ore bearing areas, due to the presence of numerous interesting species, including lichen species, generate particular environmental conditions with exceptional biocenosis representing the heritage of regional culture. the lichen species present in the studied area are one of the specialized elements of low grasslands the community violetea calaminariae, supported on soils with above average heavy metal concentrations. these habitats are listed as rare and endangered in the habitats directive (council directive 92/43/eec on the conservation of natural habitats and of wild fauna and flora) and therefore protected in most of western europe. in poland, those areas were often devastated. currently, the situation is only slightly better, but as interest of zinc-lead ore bearing areas increase, some have been included in the natura 2000 network. acknowledgements. we are greatly indebted to professor mark r. d. seaward (university of bradford) for his kindness and valuable comments for the manuscript improvement. we also thank an anonymous reviewer for very detailed remarks. part of this research was supported by the project fm eea pl 0265 (financial mechanism of european economic area). lichens of zinc-lead mines 147 references aptroot a., boom van den p. p. g. 1998. pyrenocollema chlorococcum, a new species with a chlorococcoid photobiont from zinc-contaminated soils and wood. cryptogamie, bryologie-lichénologie 19: 193–196. boom van den p. p. g. 2000. some interesting records of lichens and lichenicolous fungi from the neth-some interesting records of lichens and lichenicolous fungi from the netherlands iv. österreichische zeitschrift für pilzkunde 9:141–145. cieśliński s., czyżewska k., fabiszewski j. 2006. red list of the lichens in poland. 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kurtzman, fell 2000). based on investigations of 18s rrna and 26s rrna, some species were placed in the genus geotrichum, family dipodascaceae, order saccharomycetales, while a vast majority of other species were included into the anamorphic basidiomycota, genus trichosporon (very closely related to cryptococcus), family filobasidiaceae, order tremellales (de hoog et al. 2000). teleomorphic stages have not been recorded in species belonging to basidiomycota. sexual forms have been described in several species of ascomycota such as dipodascus capitatus de hoog, m. th. smith et guého, which is a sexual stage of trichosporon capitatum von arx (kurnatowska, kurnatowski 2006). acta mycologica vol. 48 (2): 147–153 2013 doi: 10.5586/am.2013.017 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 148 m. dynowska et al. while fungi of the genus trichosporon are euryecological organisms occurring in all climatic zones, each species prefers an ecological niche (de hoog et al. 2000). the genus mostly includes anthropophiles which prefer the skin and its products but also colonize internal ontocoenoses as well as psychrophilic soil species (t. dulcitum (berkhout) weijman, t. sporotrichoides van oorschot et de hoog), coprophilic species close to them (t. faecale guého et m. th smith), species preferring eutrophized waters (t. laibachii guého et m. th smith) or clean waters (t. aquatile hedrick et dupont), species associated trophically with cap fungi (t. laubieri (moretz) weijman) and with birds (t. gracile guého et m. th. smith) (kurtzman, fell 2000; dynowska et al. 2008b). fungi of the genus trichosporon are exogenic saprotrophs which are opportunistic organisms. while they are not causative agents of pathological changes in healthy humans where the systemic balance is stable, they can colonize the human body asymptomatically and adopt a commensal lifestyle. they can attack the host when immunodeficient (dynowska 1996; baran 1998; dynowska et al. 2011). the occurrence of fungi of the genus trichosporon in biological materials in patients hospitalized for recurring respiratory infections, including suspected tuberculosis, was investigated in this study. materials and methods swabs were collected from the skin between fingers (i) and toes (ii), and from the groin (iii) and underarms (iv) in 50 patients, 25 men and 25 women, hospitalized at the independent public complex of tuberculosis and lung diseases in olsztyn. swabs were collected for mycological analysis and procedures used in laboratory diagnostics were followed (annaisiae, ginnis and pfaller 2003; kurnatowska, kurnatowski 2006). fifty healthy persons presenting for a routine medical check-up were the control group. fungal macrocultures were conducted on solid and liquid sabouraud medium with 0.1% chloramphenicol at 37oc for 48-72 hours. the incubation period was extended to seven days for the solid medium and to three weeks for the liquid medium if fungal growth was not observed. microcultures were set up on slants coated with a thin film of nickerson agar enriched with a few drops of 1:1 broth and rabbit serum. the cultures were analyzed under a microscope after 48-72 (120) hours and the following were assessed: morphology of the hyphae, the presence of budding cells, shape, size and distribution of blastospores, arthrospores and chlamydospores. arthrospores which are the most characteristic spores of the genus trichosporon and hyphal branching were analyzed in special detail. fermentation and assimilation abilities of selected sugars and nitric compounds of the fungi were assessed (de hoog et al. 2000; kurtzman, fell 2000). keys by de hoog et al. (2000), barnett, payne and yarrow (1990), kreger – van rij (1984), kurtzman and fell (2000), kurtzman, fell and boekhout (2011) and specialist publications by baran (1998) and kurnatowska and kurnatowski (2006) were used for identification. fungi of the genus trichosporon 149 results a total of 93 isolates of fungi of the genus trichosporon (46% of the samples) were identified in hospitalized patients: 57 isolates from men (61.3%) and 36 isolates from women (38.7%). five species were detected: trichosporon asahii nishikawa shinoda, t. capitatum diddens et lodder [= magnusiomyces capitatus de hoog et m. th. smith], t. cutaneum ota [= t. beigelii vuillemin], t. inkin do carmo-sousad van uden and t. pullulans diddens et lodder [= guehomyces pullulans (lindner) fell et scorzetti] (tab. 1). considerably fewer isolates and species were recorded in the control group. a total of 52 isolates were identified (26% of the samples): 37 isolates from men (71.1%) and 15 isolates from women (28.9%). these belonged to three species: t. capitatum, t. cutaneum and t. inkin. fungi were isolated more frequently in men than in women in both groups. as many as three species were not isolated in healthy women: t. asahii, t. pullulans and t. inkin. t. asahii and t. pullulans were not isolated in healthy men (tab. 1). t. cutaneum dominated in all the subjects. the number of its isolates was comparable in the hospitalized group and the healthy subjects: 55 and 42 isolates, respectively. the number of isolates of other species was considerably higher in the patients than the number of isolates recorded in the control group. it is interesting that t. asahii, t. cutaneum and t. inkin are not included in the biosafety classification of fungi potentially pathogenic to humans (bsl-2) of the european confederation of medical mycology (de hoog 1996). discussion de hoog (2000) reports nineteen species of the genus trichosporon and specifies that nine of them can be associated with the human body. six species are of clinical importance, especially in immunocompromised persons: t. asahii, t. asteroides, t. cutaneum, t. inkin, t. mucoides and t. ovoides. three of them were isolated in table 1 fungi of the genus trichosporon isolated from hospitalized patients and from healthy persons, segregated by gender, number of isolates of individual species and isolation site: i (the skin between fingers), ii (the skin between toes), iii (groin), iv (underarms) species of fungus number of isolates persons hospitalized healthy people women men women men t. asahii 1 (i) 1 (i) t. capitatum 1 (i), 1 (ii), 3 (iii), 1 (iv) 2 (i), 5 (ii), 5 (iii), 1 (iv) 1 (ii), 1 (iii), 1 (iv) 2 (ii), 1 (iii), 2 (iv) t. cutaneum 5 (i), 5 (ii), 10 (iii), 5 (iv) 5 (i), 10 (ii), 10 (iii), 5 (iv) 4 (i), 2 (ii), 4 (iii), 2 (iv) 10 (i), 10 (ii), 5 (iii), 5 (iv) t. inkin 5 (iii) 2 (iii) t. pullulans 2 (ii), 2 (iv) 1 (i), 4 (ii), 1 (iii), 2 (iv) total 36 57 15 37 93 52 150 m. dynowska et al. our study. t. asahii is especially important in patients with acute leukemia. it is sometimes isolated from pathological changes of the skin, nails (e.g. in complicated onychomycosis, from the lymphatic system, from blood and urine and in patients with inborn defects of the immune system (de hoog et al. 2000). our results confirmed an association of fungi of the genus trichosporon with patients with a disturbed systemic and cellular homeostasis. they also showed that some species of the genus trichosporon could colonize the skin on healthy persons. according to kurnatowska and kurnatowski (2006), these fungi are associated with human keratin, which closely corresponds with our results. as well as changes within the skin and mucous membranes. kurnatowska and kurnatowski (2006) described trichosporonosis in the reproductive and urinary tracts where t. cutaneum was the aetiological agent. t. cutaneum has the broadest aetiological spectrum. it can be responsible for superficial mycosis, chronic meningitis and spreading infections in hiv carriers, systemic infections in leukaemia patients, in burn victims – after blood transfusion, and in surgical patients where catheters are often an infection route (de hoog et al. 2000). we have previously identified catheterization as an important source of mycoinfections (dynowska, biedunkiewicz 2001). although systemic infections caused by t. cutaneum are rare, their course is usually terminal. the species poses a special danger to tunic-free bedsore wounds and delayed-healing wounds, so-called „silent” post-operative infections. these wounds contain a high amount of protein in the serum and dead tissues which are a highly suitable substrate for the growth and multiplication of fungi. infected wounds are one of the sources of nosocomial infections (dynowska, góralska and rosłan 2008a). observations show that t. cutaneum is one of the dominants in the mycobiota in hospital rooms (dynowska, pacyńska and karaszewska 2008c) and it exhibits increasing expansiveness towards organ ontocoenoses in humans, especially in hospitalized patients (swoboda-kopeć et al. 2001). its prevalence in the respiratory system is high (dynowska 1993). it was often isolated from the oral cavity, sputum and bronchoscopic fluid in persons diagnosed with cancer of the respiratory system and suspected of tuberculosis (dynowska, biedunkiewicz and sucharzewska 2002). high prevalence of multifocal infections was observed in this group of patients (dynowska, ejdys and kisicka 2004). reconnaissance studies (dynowska et al. 2008b) of endoscopic material from persons with unidentified digestive system complaints show that the species has also colonized this ecological niche. t. capitatum is listed by kurtzmann and fell (2000) as a species that can colonize the oral cavity and the digestive tract. it was isolated relatively frequently from the respiratory tract (dynowska 1993). t. capitatum was detected in the sputum and on the skin of healthy humans but it may be an aetiological factor of terminal systemic mycosis (baran 1998). it has been shown to occur at different sites on the skin in healthy humans (except hands). our results confirmed that t. pullulans rarely occurred on the skin and it was isolated only from patients. it is mostly reported from the oral cavity and different sections of the respiratory tract (dynowska 1993, 1996). it accompanies t. cutaneum in our study. t. inkin, reported as a potentially pathogenic species only in the polish literature (kurnatowska, kurnatowski 2006), is especially interesting. it was recorded in fungi of the genus trichosporon 151 hospitalized and healthy men in our study. it seems to be gender-specific and associated with men. this is also suggested in studies by other authors (lopes et. al. 1997; guého et al. 1994) who stressed its broad aetiological spectrum: from dermatomycosis, especially of damaged skin, to infections of the urinary tracts and terminal sections of the digestive tract, to endocarditis. drutz (1986) emphasized the risk of infection of organ ontocoenoses by t. cutaneum, t. capitatum and t. pullulans as early as over 20 years ago. according to drutz (1986), these species are widespread in the biosphere and are a frequent component of the skin mycobiota. they can penetrate the oral and nasal cavities from the skin and transfer into to the ontosphere from there. it is disconcerting that as many as three species of the five species isolated by us are classified as bsl-2. this category comprises fungi colonizing organisms of invertebrates and vertebrates, with a high ability to survive in tissues of different organs, that can cause deep-seated opportunistic infections in immunocompromized persons (de hoog 1996). according to annaisiae et al. (2003), fungi of the genus trichosporon prefer sites around the groin, sexual organs and the anus. piedra alba, a chronic infection of the hair in different parts of the body, as well as infections of nails and mucous membranes of the oral cavity are very common infections caused by fungi of the genus trichosporon, mostly t. cutanum and t. inkin, in the tropical and subtropical climates (richardson, warnock 1993). due to intensive tourism these diseases and their causal agents can be unintentionally introduced to our climatic zone. the risk of infection by new species of fungi increases as they widen the area of their occurrence in the biosphere and penetrate the human ontosphere (dynowska 1996; dynowska, biedunkiewicz 2001; dynowska et al. 2008b). conclusions human skin should be treated as a natural reservoir of potentially pathogenic fungi. as well as being inhabited by typical dermatophytes, it is colonized by numerous yeasts. fungi of the genus trichosporon seem to be of primary importance. references annaisiae e.j., mc ginnis m.r., pfaller m.a. 2003. clinical mycology. els. scien. and church. leving, new york, edinburgh, london, philadelphia. baran e. 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(in:) j. garbacz (ed.). diagnozowanie stanu środowiska. prace komisji ekologii i ochrony środowiska btn: 47-54. dynowska m., góralska k., troska p., barańska g., biedunkiewicz a., ejdys e., sucharzewska e. 2011. results of long-standing mycological analyses of biological materials originating from selected organ ontocenoses-yeast and yeast-like fungi. wiad. parazyt. 57 (2): 97-102. guého e., improvisi g. s., de hoog g. s., dupont b. 1994. trichosporon on humans: a practical account. mycoses 37: 3-10. http://dx.doi.org/10.1111%2fj.1439-0507.1994.tb00277.x kurnatowska a., kurnatowski p. 2006. mikologia medyczna. promedi łódź. kurtzman c.p., fell j.w. 2000. the yeast. a taxonomic study. 4th ed., elsevier – amsterdam. kurtzman c.p., fell j.w., boekhout t. 2011. the yeast. a taxonomic study. 5th ed. elsevier – tokyo. lopes j.o., alves s.h., klock c., oliveira t.l.o., forno n.r.f. 1997. trichosporon inkin peritonitis during continuous ambulatory peritoneal dialysis bibliography review. mycopathol. 139: 15-18. http:// dx.doi.org/10.1023%2fa%3a1006870017725 richardson m.d., warnock d.w. 1993. fungal infection. diagnosis and management. blackwell scien. publ. – oxford. swoboda-kopeć e., rakosz a., sawicka-grzelak a., wróblewska m., krawczyk e., stelmach e., łuczak m. 2001. etiologiczne czynniki fungemii u hospitalizowanych pacjentów. med. dośw. mikrobiol. 53: 291-295. grzyby z rodzaju trichosporon izolowane ze skóry osób hospitalizowanych streszczenie grzyby z rodzaju trichosporon są szeroko rozpowszechnione w biosferze i stanowią częsty komponent mykobioty skóry (powinowactwo do keratyny ludzkiej). z niej mogą dostawać się do jamy ustnej, jam nosa a następnie przenosić się w głąb ontosfery. jako formy oportunistyczne u osób zdrowych prowadzą komensaliczny tryb życia, u osób z zaburzeniami odporności mogą wywoływać zmiany chorobowe (trychosporonozy) o różnym nasileniu i różnym zasięgu. oprócz skóry i błon śluzowych najczęściej atakowany jest układ moczowy i płciowy a jako droga zakażenia wymieniane są cewniki. odnotowano przypadki zakażeń układowych kończących się śmiercią. grzyby z rodzaju trichosporon są szczególnie niebezpieczne dla trudno gojących się ran chirurgicznych i ran odleżynowych. materiał badawczy stanowiły wymazy ze skóry między palcami rąk i nóg oraz pachwin i pach 50 pacjentów (25 kobiet i 25 mężczyzn) spzg i chp w olsztynie (hospitalizacja z powodu nawracających infekcji układu oddechowego, w tym podejrzeniami o gruźlicę). grupą kontrolną było 50 osób zdrowych, zgłaszających się na badania okresowe. analizy laborato fungi of the genus trichosporon 153 ryjne i oznaczanie grzybów do gatunku przeprowadzono zgodnie z ogólnie przyjętym tokiem diagnostycznym w laboratoriach mykologicznych. od pacjentów hospitalizowanych uzyskano 93 izolaty grzybów z rodzaju trichosporon należące do 5 gatunków: t. asahii, t. capitatum, t. cutaneum. t. inkin i t. pullulans. w grupie kontrolnej uzyskano 52 izolaty reprezentujące 3 gatunki: t. capitatum, t. cutaneum i t. inkin. w obydwu grupach grzyby częściej notowano u mężczyzn niż u kobiet. zdecydowanym dominantem był t. cutaneum – liczba jego izolatów u osób hospitalizowanych i zdrowych była porównywalna. w przypadku pozostałych gatunków liczba izolatów uzyskanych od osób hospitalizowanych znacznie przewyższała liczbę izolatów stwierdzonych w grupie kontrolnej. wszystkie gatunki odnotowane w przeprowadzonych badaniach należą do potencjalnych antropopatogenów, a t. asahii, t. cutaneum i t. inkin są uwzględnione w klasyfikacji biobezpieczeństwa z kategorią bsl-2. w tej kategorii znajdują się grzyby zasiedlające organizmy bezkręgowców i kręgowców, o dużej zdolności przeżywania w tkankach różnych narządów, mogące wywoływać głębokie oportunistyczne zakażenia. reasumując, skórę człowieka należy traktować jako jeden z naturalnych rezerwuarów mikrogrzybów potencjalnie chorobotwórczych. oprócz typowych dermatofitów jest ona siedliskiem licznych drożdży, wśród których grzyby z rodzaju trichosporon wydają się odgrywać pierwszoplanową rolę. 2013-12-20t14:40:09+0100 polish botanical society new data on some rare species of hyphomycetes from lithuania svetlana markovskaja institute of botany, laboratory of mycology žaliųjų ežerų 49, lt–2021 vilnius, svetlana.m@botanika.lt m a r k o v s k a j a s.: new data on some rare species of hyphomycetes from lithuania. acta mycol. 42 (1): 93-98, 2007. the paper presents new data on ecological peculiarities of five anamorphic fungi species, cladorrhinum foecundissimum, dwayaangam cornuta, lateriramulosa uni-inflata, pyramidospora herculiformis and tetraploa setifera, their distribution and substrate preferences are discussed. short description, illustration and map of localities in lithuania of recorded species are presented. key words: anamorphic fungi, hyphomycetes, diversity, ecology, distribution, lithuania introduction anamorphic fungi inhabiting forest litter are characterized by wide adaptation abilities in different environments, but their species composition and ecology are still insufficiently known. the best-known and most studied are terrestrial, aquatic and aero-aquatic groups of hyphomycetes (e l l i s 1971, 1976; m a t s u s h i m a 1975; i n g o l d 1975; b ä r l o c h e r 1992 et al.). during the studies of hyphomycetes associated with forest litter in various terrestrial and freshwater ecosystems in mixed deciduous forests of central and southeast lithuania in 1999–2004, five rare and interesting for their ecological adaptation patterns species were obtained both in terrestrial and aquatic habitats. recorded species were mainly polytrophic and developed as saprobes on dead woody plant substrates. till present time three reported here fungi species, cladorrhinum foecundissimum, lateriramulosa uni-inflata and tetraploa setifera were known exceptionally as terrestrial or soil-inhabiting ( m a t s u s h i m a 1971; m o u c h a c c a , g a m s 1993; r é v a y 1993), other two, dwayaangam cornuta and pyramidospora herculiformis as aquatic ( d e s c a l s , we b s t e r 1982; s i n g h 1976). our observation indicated that they can grow, sporulate and actively decompose plant debris of forest litter both in moist terrestrial and submerged conditions. whereas these species are characterized by amphibious nature of development, it is best to place them into the specific and still acta mycologica vol. 42 (1): 93-98 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 94 s. markovskaja poorly studied ecological group of “amphibious” fungi (m i c h a e l i d e s , k e n d r i c k 1978; b ä r l o c h e r 1992; d i x , we b s t e r 1995) and regard them as such. part of the reported here species are known only from few collections worldwide and our data supplement knowledge on their substratum specificity, habitats and geography. materials and methods the specimens were collected directly from the terrestrial forest litter or were obtained by cultivating terrestrial litter in moist chamber in the laboratory. submerged leaves were collected from aquatic habitats (forest streams, lakes, rivers) and later incubated in petri dishes under the thin layer of distilled water. the fungi were studied using standard microscopic techniques and identified basing on their morphological properties and character of conidiogenesis. descriptions and illustration were made from fresh preparations in distilled water. specimens of dried material and permanent glass slides with lacto-phenol are deposited in the fungal collection at the herbarium of the institute of botany (bilas), vilnius. description and discussion cladorrhinum foecundissimum sacc. & marchal, bull. soc. r. bot. belg., 24: 65, 1885. (fig. 1 a). mycelium yellowish-grey with clusters of subhyaline, branched, 3–4 μm wide, vegetative hyphae. conidiogenous cells integrated, intercalary, monophialidic, deconidiogenous cells integrated, intercalary, monophialidic, determinate. phialides subhyaline, ampulliform or subspherical with distinct collarettes, 6.3–9 x 1.5–3.6 μm. conidia subhyaline or hyaline, smooth-walled, dacryoid or obclavate, 2.5–2.7 μm in diameter, aggregated in slimy masses. material examined: on decaying unidentified wood, moist forest litter, kėdainiai distr., lančiūnava forest, september 21, 1999; on submerged unidentified wood, lazdijai distr., lake ragožis, august 30, 2001. note: in europe cladorrhinum foecundissimum previously has been isolated mainly from soil (a r x , g a m s 1966; h o l u b o v á j e c h o v á 1979; b o r o w s k a 1986; m o u c h a c c a , g a m s 1993). from submerged conditions it was noted for the first time by h y d e and g o h (1999) on unidentified wood in the u.k. in lithuania it was collected both from terrestrial and submerged habitats. accordingly this species could be placed into the group of “amphibious” fungi. this species is widely distributed in northern hemisphere, though uncommon. lateriramulosa uni-inflata matsush., microfungi of the solomon islands and papua-new guinea (osaka): 34, 1971. (fig. 1 b). mycelium hyaline. conidiophores semi-micronematous, hyaline, up to 20 μm long. conidiogenous cells holoblastic, terminal, determinate, integrated. conidia hyaline, composed of three indentate branches, 20–23 μm in diameter. material examined: on acorns of quercus robur, moist forest litter, vilnius environs, verkiai regional park, november 3, 1999; on submerged leaves of tilia cordata, vilnius environs, verkiai regional park, forest stream, november 12, 2001; on hyphomycetes from lithuania 95 submerged leaves of quercus robur, vilnius environs, verkiai regional park, river neris, november 12, 2001. note: this species was described by m a t s u s h i m a (1971) on decaying leaves as terrestrial fungus from papua-new guinea and later was also collected in japan fig. 1. a – hyphae with phialides and conidia of cladorrhinum foecundissimum; b – conidia of lateriramulosa uni-inflata; c – conidiophore with conidia of dwayaangam cornuta; d – conidia of pyramidospora herculiformis; e – conidia of tetraploa setifera. scale bars: a = 10 µm; b-e = 20 µm. 96 s. markovskaja (m a t s u s h i m a 1975). it is also known from north america (c r a n e 1968). in europe it is rarely collected as waterborne from washed litter and foam (m a r v a n o v á 1973; i n g o l d 1975; g ö n g z ö l , r é v a y 1983). in lithuanian specimens l. uniinflata sporulated abundantly after incubating submerged leaves for several days in petri dishes under a thin layer of distilled water. probably the fungus is not genuinely waterborne, but amphibious and is likely to be cosmopolitan in its distribution. dwayaangam cornuta descals, trans. br. mycol. soc. 1982, 78 (3): 408 – 411. (fig. 1 c). mycelium hyaline. conidiophores semi-micronematous, hyaline, 10–12 x 4 μm. conidiogenous cells holoblastic, terminal, determinate. conidia hyaline, 40–75 x 30–50 μm, composed from main axis of 1–4 cells, 10–30 x 4–6 μm and four branches of 1–4–5 cells, in pairs, (17) 23–35 x 4.5–6.5 μm. material examined: on decaying wood of acer platanoides l., moist forest litter, vilnius environs, verkiai regional park, november 4, 1999; on decaying wood of quercus robur, moist forest litter, kėdainiai distr., juodkiškiai forest, october 3, 2001; from foam, trakai distr., jagelonys forest, forest stream, november 14, 2004. note: d e s c a l s described this species as aquatic after incubating a piece of wood (observed specimen was collected in streamlet in u.k.) under the thin layer of water (d e s c a l s , we b s t e r 1982), but earlier in u. k. conidia of this fungus were frequently observed in foam as unidentified (i n g o l d 1942,1959, 1975). in lithuania this species was collected twice from moist forest litter on decaying wood and once from foam, thus it could be characterised as amphibious. probably it is widely distributed in the temperate zone of northern hemisphere. pyramidospora herculiformis n. singh, trans. br. mycol. soc. 1976, 66 (2): 347 – 350. (fig. 1 d). mycelium hyaline. conidiophores semi-macronematous, simple, 20–25 μm long, conidiogenous cells holoblastic, terminal, determinate. conidia are h-shaped, composed from main axis with 2 cells and four lateral branches of 2–3 (4) cells, 20–25 x 6–8 μm in length. material examined: on submerged leaves of corylus avellana l., varėna distr., merkinė environs, dzūkija national park, strauja geomorphologic reserve, river strauja, november 1, 1997; on submerged leaves of corylus avellana l., kėdainiai distr., barupė hydrological reserve, river barupė, september 21, 1999; on fallen leaves of coryllus avellana, moist forest litter, kėdainiai distr., stebuliai forest, may 6, 1999. note: this species was described by s i n g h (1976) as an aquatic hyphomycete collected on submerged rotting leaves of unidentified dicotyledonous plants from africa (sierra leone). in lithuania this fungus was collected on both submerged leaves of corylus avellana and leaves of the same plant from moist forest litter, thus this species should be ascribed to amphibious fungi. here it is for the fist time recorded for europe. tetraploa setifera ágnes révay, nova hedwigia, 1993, 56 (3-4): 480 – 481. (fig.1 e). mycelium pale brown. conidiogenous cells holoblastic, intercalary, integratet, determinate. conidia sessile. the body of conidia are dark brown, smooth, com hyphomycetes from lithuania 97 posed of four closely developing columns of 4–5 (6) cells, 17–30 x 12–13 μm with appendages. appendages hyaline, smooth, 3–4 septate, 25–100 x 4–2 μm. material examined: on decaying wood of fraxinus excelsior l., moist forest litter, kėdainiai distr., berunkiškiai forest, october 2, 2002. note: present collection of tetraploa setifera is the first record of this species from lithuania and the second record in the world. originally for the first time this species was described from hungary on rotten unidentified wood collected near the stream from moist habitat ( r é v a y 1993). lithuanian specimen morphologically (by shape and size of conidia) is practically identical to the published description of type material. among six species of genus tetraploa, only tetraploa setifera always produce smooth-walled conidia with closely adpressed columns in the apical part. morphologically t. setifera is very close to commonly found on grasses t. aristata berk. & br., which in hungary was also collected on submerged wood of fagus sylvatica ( r é v a y 1993). probably future ecological observation might confirm that t. aristata and t. setifera both belong to group of amphibious fungi. until present t. aristata is known only from europe. the fact of record of such rare species as cladorrhinum foecundissimum, dwayaangam cornuta, lateriramulosa uni-inflata, pyramidospora herculiformis and tetraploa setifera in different terrestrial and submerged habitats in lithuania, suggests that distribution and ecology of these fungi is far wider than was reputed. the distribution of above-mentioned species in lithuania is illustrated in figure 2. fig. 2. localities of cladorrhinum foecundissimum (♦), lateriramulosa uni-inflata (▲), dwayaangam cornuta ( ■ ), pyramidospora herculiformis (●) and tetraploa setifera (★) in lithuania. ■★  ♦●● ▲■  ▲▲ ♦ ■ ● 98 s. markovskaja acknowledgements: the author would like to extend a sincere gratitude to the polish colleagues for kind invitation to contribute to this special issue. financial support by the lithuanian state science and stud-financial support by the lithuanian state science and studies foundation for the studies is kindly acknowledged. references a r x j . a., g a m s w. 1966. über pleurage verruculosa und die zugehörige cladorrhinum konidenienform. nova hedwigia 13: 199–208. b ä r l o c h e r f. 1 9 9 2 . the ecology of aquatic hyphomycetes. ecological studies 94: 1–225. b o r o w s k a a . 1986. flora polska. grzyby (mycota) 16: grzyby niedoskonałe deuteromycetes). pwn. warszawa–kraków. c r a n e j . l . 1 9 6 8 . freshwater hyphomycetes of the northern appalachian highland including new england and three coastal plain states. am. j. bot. 55: 996–1002. d e s c a l s e . , we b s t e r j . 1982. taxonomic studies on aquatic hyphomycetes. iii. some new species and a new combinations. trans. br. mycol. soc. 78 (3): 405–437. d i x n . j . , we b s t e r j . 1995. fungal ecology. chapman & hall. london. e l l i s m. b. 1971: dematiaceous hyphomycetes. kew. e l l i s m. b. 1976: more dematiaceous hyphomycetes. kew. g ö n c z ö l j . , r é v a y á . 1983. observations on the hyphomycetes inhabiting forest litter in hungary. acta bot. hungarica 29 (1-4): 107–125. h o l o b o v á j e c h o v á v. 1979. lignicolous and some other saprophytic hyphomycetes from hungary. česká mykologie 33: 138–149. h y d e k . d ., g o h t. k .1999. fungi on submerged wood from river coln, england. mycol. res. 103 (12): 1561–1574. m a r v a n o v á l. 1973. notes on lateriramulosa uni-inflata. trans. br. mycol. soc. 60: 145–147. m a t s u s h i m a t. 1971. microfungi of the sodomon islands and papua-new guinea. kobe. m a t s u s h i m a t. 1975. icones microfungorum a matsushima lectorum. japan, shinogi & co., ltd, kobe. m i c h a e l i d e s j . , k e n d r i c k b . 1978. an investigation factors retarding colonization of conifer needles by amphibious hyphomycetes in streams. mycologia 70: 419–430. m o u c h a c c a j., g a m s w. 1993. the hyphomycete genus cladorrhinum and its teleomorph connection. mycotaxon 48: 415–440. r é v a y a. 1993. some new or interesting hyphomycetes from hungary. nova hedwigia 56 (3-4): 473– 482. i n g o l d c . t. 1942. aquatic hyphomycetes of decaying alder leaves. trans. br. mycol. soc. 25: 339– 417. i n g o l d c . t. 1959. submerged aquatic hyphomycetes. journal of the quekett microscopical club 4-5: 115–130. i n g o l d c . t. 1975. an illustrated guide to aquatic and waterborne hyphomycetes (fungi imperfecti) with notes on their biology. freshwater biological association scientific publication 30: 1–96. s i n g h n. 1976. pyramidospora herculiformis sp. nov., a new aquatic hyphomycete from sierra leone. trans. br. mycol. soc. 66 (2): 347–350. nowe dane o kilku rzadkich na litwie gatunkach hyphomycetes s t r e s z c z e n i e praca zawiera nowe dane do charakterystyki ekologicznej pięciu gatunków grzybów anamorficznych: cladorrhinum foecundissimum, dwayaangam cornuta, lateriramulosa uniinflata, pyramidospora herculiformis i tetraploa setifera. dyskutowane są ich preferencje co do substratu oraz podane są ich cechy morfologiczne i rozmieszczenie stanowisk poszczególnych gatunków na mapie litwy. 2014-01-01t11:45:28+0100 polish botanical society assessment of in vitro antifungal activity of preparation “fin candimis” against candida strains anna głowacka1, anna bednarek-gejo1, danuta trojanowska1, mariusz mianowany1 and alicja budak2 1environmental biology unit, medical university of lodz gen. hallera 1, pl-90-647 łódź, anna.glowacka1@umed.lodz.pl 2pharmaco-microbiology unit, collegium medicum jagiellonian university, medyczna 9, pl-30-688 kraków głowacka a., bednarek-gejo a., trojanowska d., mianowany m., budak a.: assessment of in vitro antifungal activity of preparation “fin candimis” against candida strains. acta mycol. 47 (1): 27–34, 2012. the aim of the study was to assess the antifungal activity of preparation „fin candimis” (oregano essential oil) against yeast-like strains belonging to the genus candida. during the investigation, there were used up nine candida albicans strains and ten c. glabrata strains isolated from different clinical material, along with one c. albicans demonstration strain atcc 90028. the oregano essential oil, utilized in the study, was obtained from fresh leaves of origanum vulgare l. and bore a trade name „fin candimis”. according to data yielded by its manufacturer, concentration of pure oregano essential oil in preparation „fin candimis” totals up to 210 mg/ml. the susceptibility of the candida strains to preparation „fin candimis” was assessed by means of the disc-diffusion method, upon the sabouraud solid medium (after a 24-hour incubation of the cultures at temperature of 37 degrees centigrade); the oregano essential oil had been diluted in 1 ml of dmso, according to the geometrical progression. a measure of the antifungal activity of preparation „fin candimis” was the minimal inhibitory concentration (mic), in terms of the fungus growth. preparation „fin candimis” is capable of being applied in the prevention and treatment of candidiasis – alone, or as a natural adjunctive agent. the c. albicans strains are more susceptible to preparation „fin candimis” in comparison to the c. glabrata ones. key words: candida albicans, c. glabrata, oregano essential oil, susceptibility acta mycologica vol. 47 (1): 27–34 2012 28 a. głowacka et al. introduction according to epidemiological data spreading over the past two decades, there has been observed in the world a significant increase in the number of infections caused by fungi belonging to the genus candida, referring particularly to patients admitted to intensive care units, undergoing immunosuppression or taking broad-spectrum antibiotics, and after having received a graft (rex 1995; eggimann, gabrino and pit-eggimann, gabrino and pitter 2003; krutkiewicz 2010). the epidemiological situation impels scientists to search for new therapeutic agents – among the others, natural and vegetal ones – which bear the stamp of high effectiveness and low toxicity. these compounds could be useful in the field of antifungal prophylaxis and treatment. to this group there belong essential oils which present antifungal properties – inclusive of origanum vulgare essential oil, popularly named oregano. the name oregano has been derived from two greek words: oros – „a mountain”, and ganos – „an adornment”; it stands for „the mountain’s joy”, and is connected with the area the plant overgrows in central and southern europe. for many ages, origanum vulgare has been known for its healing properties. by the first century a.d., greek physician pedanius dioscorides, and, before him, hippocrates (460–370 b.c.), recognized the father of contemporary medicine, along with philosopher and erudite aristotle (384–322 b.c.), had appreciated oregano as a powerful antidote to various sorts of maladies. the ancient medicals customarily used o. vulgare as an analgesic, carminative, disinfecting, and detoxifying drug. o. vulgare was widely used also in folk medicine; it was administered in combination with wine, e.g., to individuals who had been bitten by animals. in the middle ages, oregano was an ingredient of the infusion that was dispensed in order to protect from wizardries. nowadays, o. vulgare did not forfeit its remedial purport, none the less the plant is predominantly associated with a condiment which is widely used within the mediterranean cuisine. in support of the oregano’s numerous beneficial properties, which had been observed in the past ages, there were adduced reliable results due to the plentiful scientific investigations. the biological activity of the oregano essential oil (obtained from its fresh leaves) is closely connected with its phenol compound content (it gives a distinctive taste and flavour). o. vulgare comprises up to 3% of the essential oil which is fruitful in phenols such as carvacrol and thymol. the phenolic content comes up even to 60%. oregano contains also sesquiterpenes, catechol, phenolic acids and flavonoids (hołderna-kędzia 2010). the aim of the study was to assess the antifungal activity of preparation „fin candimis” (oregano essential oil) against yeast-like strains belonging to the genus candida, isolated from different clinical materials, and against a candida albicans demonstration strain atcc 90028. assessment of in vitro antifungal 29 material and methods in the present study, there were subjected to investigation 20 strains of fungi belonging to the genus candida: 10 c. albicans strains and 10 c. glabrata ones. the strains derived from patients who had been hospitalized in the ludwik rydygier specialistic hospital in kraków in the following departments: intensive care unit, cardiologic dept., haematologic dept., orthopaedic dept., internal diseases dept., and rehabilitation unit. the investigated strains were isolated from aspirate of the upper airways (18 strains), from urine (one strain), and one strain constituted a reference pattern, i.e., c. albicans atcc 90028. the oregano essential oil, which was employed in the study, had been obtained from the o. vulgare fresh leaves, and bore a trade name „fin candimis”. the oregano essential oil issued from the fin club poland co. ltd., and was elaborated by the business concern named hankintatukku oy in karkkila (finland). according to the manufacturer’s data, concentration of the o. vulgare pure essential oil in preparation „fin candimis” amounts to 210 mg/ml; the essential oil contains from 75% to 85% of carvacrol (chemical name: 5-isopropyl-2-methylphenol; molecular formula: c10h14o). the investigated candida strains’ susceptibility to preparation „fin candimis” was assessed by means of the disc-diffusion method on an agar medium (agar gel). the isolated fungal strains were inoculated upon sabouraud medium and exposed to a 24-hour incubation at temperature 37 degrees centigrade. from the cultures, there was prepared an inoculum in sterile physiological saline, given density 0,5 on the mcfarland scale. the suspensions of the investigated candida strains, prepared as above, in a volume of 100 μl, were inoculated upon sabouraud medium, whereafter they were fixed in the medium discs made of blotting-paper, each one comprising 15 μl of preparation „fin candimis” (oregano essential oil) in a required concentration (the oregano essential oil had been diluted in 1 ml of dimethyl sulfoxide – dmso, according to the geometrical progression). the petri dishes were incubated over a 24-hour period in a thermostat at temperature of 37 degrees centigrade, whereupon there were measured diameters of the growth inhibition zones around the discs, n = [mm], in case of each investigated fungal strain, assuming a mean value from two diameters being perpendicular to each other. the minimal inhibitory concentration, mic = [mg/ml], was obtained from a growth curve that had been drawn in a semi-logarithmic cartesian coordinate system, i.e., apposing: on the abscissa – a common logarithm of concentration of preparation „fin candimis”, and on the ordinate – a mean diameter of the growth inhibition zone, n = [mm], after the 24-hour incubation. a measure of the antifungal activity of preparation „fin candimis” was its minimal inhibitory concentration (mic), in terms of the fungus growth, which was computed after the kadłubowski’s (1971) modified formula: where: — n1, — n2 – arithmetical means, obtained for two groups of diameters of the growth inhibition zones, — log1 c, — log2 c – arithmetical means for the common logarithm, computed for two groups of concentrations of the preparation. 30 a. głowacka et al. in order to estimate statistically the diameters of the growth inhibition zones there were computed: mean values, standard errors of mean, standard deviations, confidence intervals, along with minimal and maximal values (petrie, sabin 2009). results the antifungal activity of preparation „fin candimis” was assessed in vitro by means of the disc-diffusion method against 19 fungi strains belonging to the genus candida, which had been isolated from aspirate of the upper airways (18 strains) and from urine (one strain). from amongst 19 candida strains, nine strains belonged to the species c. albicans, and ten did to the species c. glabrata. moreover, there was determined the antifungal activity of preparation „fin candimis” against the c. albicans demonstration strain atcc 90028. diameters of the growth inhibition zones, n = [mm], in case of the c. albicans clinical strains exposed to the activity of five concentrations of the oregano essential oil, according to the concentration adhibited, amounted to 44-55 mm at the outmost, given concentration 210 mg/ml (on the average 48.78 ± sd = 4.12 mm), and minimally to 8-10 mm, given concentration 13.125 mg/ml (on the average 9.11 ± sd = 0.93 mm). diameters of the growth inhibition zones, n = [mm], for the c. albicans demonstration strain atcc 90028 exposed to the activity concentrations of the oregano essential oil amounted to 47 mm and 10 mm, respectively (tab. 1). diameters of the growth inhibition zones, n = [mm], in case of candida glabrata strains exposed to the activity of five concentrations of the oregano essential oil, according to the concentration, amounted to 22-45 mm at the outmost (on the average 36.00 ± sd = 7.72 mm), given concentration 210 mg/ml, and minimally to 6-10 mm (on the average 7.60 ± sd = 1.07 mm), given concentration 13.125 mg/ml (tab. 2). table 1 growth inhibition zones n = [mm] for nine candida albicans strains isolated from the patients and one candida albicans demonstration strain under the influence of different concentrations of preparation “fin candimis” sn species strain number clinical material concentration of the oregano essential oil [mg/ml] 210 105 52.5 26.25 13.125 growth inhibition zone, n = [mm] 1. c. albicans atcc 90028 demonstration strain 47 30 28 14 10 2. c. albicans 724/09 aspirate 45 30 23 12 8 3. c. albicans 733/09 aspirate 45 30 28 11 10 4. c. albicans 765/09 aspirate 45 33 22 10 8 5. c. albicans 893/09 aspirate 50 32 27 15 10 6. c. albicans 956/09 aspirate 53 35 30 17 9 7. c. albicans 1164/09 aspirate 44 33 20 11 9 8. c. albicans 340/10 aspirate 52 35 30 15 10 9. c. albicans 369/10 aspirate 55 40 30 10 8 10. c. albicans 21/11 urine 50 40 30 12 10 assessment of in vitro antifungal 31 the minimal inhibitory concentration (mic) values for preparation „fin candimis” in case of nine investigated c. albicans and ten c. glabrata clinical strains, isolated from the hospitalized patients, figured out at 13.38 mg/ml and 13.58 mg/ml, respectively. the statistical computations for the growth inhibition zones n = [mm] in case of nine c. albicans and ten c. glabrata strains exposed to different concentrations of preparation „fin candimis” were submitted to tables 3 and 4. the mean diameters table 2 growth inhibition zones n = [mm] for ten candida glabrata strains isolated from the patients under the influence of different concentrations of preparation “fin candimis” sn species strain number clinical material concentration of the oregano essential oil [mg/ml] 210 105 52.5 26.25 13.125 growth inhibition zone, n = [mm] 1. c. glabrata 372/09 aspirate 42 28 23 12 8 2. c. glabrata 523/09 aspirate 35 25 20 8 6 3. c. glabrata 627/09 aspirate 28 32 22 10 7 4. c. glabrata 669/09 aspirate 45 33 22 10 8 5. c. glabrata 762/09 aspirate 40 30 23 10 8 6. c. glabrata 768/09 aspirate 22 30 22 12 8 7. c. glabrata 981/09 aspirate 45 37 17 8 7 8. c. glabrata 1041/09 aspirate 33 28 22 10 7 9. c. glabrata 10/11 aspirate 40 30 23 15 10 10. c. glabrata 393/10 aspirate 30 25 23 14 7 table 3 statistical computations for the growth inhibition zones n = [mm] for nine candida albicans strains isolated from the patients under the influence of different concentrations of preparation “fin candimis” concentration [mg/ml] mean standard deviation standard error 95% confidence interval minimum maximum s.n. 1. 210.000 48.78 4.12 1.37 45.61–51.94 44 55 2. 105.000 34.22 3.73 1.24 31.35–37.09 30 40 3. 52.500 26.67 3.97 1.32 23.62–29.72 20 30 4. 26.250 12.56 2.51 0.84 10.63–14.48 10 17 5. 13.125 9.11 0.93 0.31 8.40–9.82 8 10 table 4 statistical computations for the growth inhibition zones n = [mm] for ten candida glabrata strains isolated from the patients under the influence of different concentrations of preparation “fin candimis” concentration [mg/ml] mean standard deviation standard error 95% confidence interval minimum maximum s.n. 1. 210.000 36.00 7.72 2.44 30.48–41.52 22 45 2. 105.000 29.80 3.65 1.15 27.19–32.41 25 37 3. 52.500 21.70 1.89 0.60 20.35–23.05 17 23 4. 26.250 10.90 2.33 0.74 9.23–12.57 8 15 5. 13.125 7.60 1.07 0.34 6.83–8.37 6 10 32 a. głowacka et al. of the growth inhibition zones for the c. albicans clinical strains are larger than the corresponding mean values in case of the c. glabrata strains, given all the five adhibited concentrations of the investigated oregano essential oil. discussion the susceptibility of microorganisms to essential oils is an outcome of the microorganisms’ peculiarities, and the essential oils’ properties. within the antifungal activity of the essential oils, two factors play the leading role: lipophilous character of the hydrocarbon skeleton, and hydrophilous nature of the functional groups. the sequence of the essential oils’ properties appears as follows: phenols > aldehydes > ketones > alcohols > esters > ethers > hydrocarbons. the phenols – thymol, carvacrol and eugenol – demonstrate the strongest antimicrobial activity. the explanation of this phenomenon is the alkaline character of the hydroxylic group that may set up hydrogen bonds with the enzyme’s active site (knobloch et al. 1989). therefore the essential oils, which are mostly composed of phenols, exert themselves most powerfully upon the microorganisms, and possess the broadest spectrum in terms of their antimicrobial activity. various studies conducted in order to uncover the correlation between the composition of the essential oils and their specific activity has as yet come to naught (pattnaik 1997). it is possible that not the essential oil particular contents, but their mutual proportions do determine the essential oil’s activities (kalemba 1999). concerning the practical use of many essential oils, it is crucial to attain a fast effect, and to protract the essential oil’s antimicrobial activity (akgiil, kivanc 1989). inconsistent ways in which various scientific investigators tend to present the mic values do trammel their likening. a similar hitch is the diversity regarding a range of concentrations of the essential oils, along with discrepant methods of measurement. when comparing outcomes of various studies, it occurs that one ought to standardize the units (kalemba 1999). in the present paper, in order to preserve the integrity of results, the mic values are expressed in the units which have been stated by the authors of cited articles. kursat and erecevit (2008) proved the germicidal activity of oregano essential oil against bacteria pseudomonas aeruginosa, klebsiella pneumoniae, staphylococcus aureus and bacillus megaterium, and fungi belonging to the genus epidermophyton. yet they did not demonstrate the antifungal activity against candida and trichophyton strains. during their research work, the authors made use of essential oil obtained from the plant named origanum vulgare l. subsp. gracile (c. koch). this essential oil, diluted in 98.1% solution of methyl alcohol, showed the germicidal activity against klebsiella pneumoniae, staphylococcus aureus and bacillus megaterium. the study conducted in spain by lópez et al. (2007) showed that the oregano essential oil displayed its antifungal properties. it exerted an effect upon candida albicans, apsergillus flavus and penicillium islandicum strains, along with gram-positive bacteria (staphylococcus aureus, enterococcus faecalis, listeria monocytogenes, bacillus cereus) and gram-negative ones (escherichia coli, psuedomonas aeruginosa, assessment of in vitro antifungal 33 yersinia enterocolitica i salmonella choleraesuis). the mic value in case of the candida albicans strains amounted to 17.5 μl/l (lópez et al. 2007). consecutive studies, aiming at estimating the susceptibility of candida strains, were carried out in the united states by manohar et al. (2001). the researchers proved that the oregano essential oil inhibited the growth of the investigated candida albicans strains. the mic value, provided by the authors, figured out at 0.125 mg/ml. the discussed study was conducted by the use of one candida albicans demonstration strain atcc 48274, and strains that stemmed from manohar et al.’s own culture. the authors administered the oregano essential oil to mice infected with candidiasis; the essential oil was prepared in controlled doses: the higher was the dose, the more potent was the antifungal activity of the oregano essential oil (manohar et al. 2001). conclusions 1. preparation „fin candimis” (oregano essential oil) is capable of being applied in the prevention and treatment of candidiasis – alone, or as a natural adjunctive agent. 2. the investigated candida albicans strains are more susceptible to preparation „fin candimis” in comparison to the candida glabrata ones. references akgiil a., kivanc m. 1989. chemical composition and antimicrobial effect of turkish laurel leaf oil. j. essent. oil res. 1: 119–128. eggimann a., gabrino j., pitter d. 2003. epidemiology of candida species infections in critically ill nonimmunosuppressed patients. infect. dis. 3: 685–701. hołderna-kędzia e. 2010. działanie substancji olejkowych na bakterie i grzyby. postępy fitoterapii. 1: 3-8. kadłubowski r. 1971. approximate methods in analysis of dose-response curve. abstracts of viith international congress of chemotherapy, praha. a-12:28. kalemba d. 1999. przeciwbakteryjne i przeciwgrzybowe właściwości olejków eterycznych. postępy mikrobiol. 38 (2): 277–280. knobloch k., pauli a., iberl b., weigand h., weis n. 1989. antibacterial and antifungal properties of essential oil components. j. essent. oil res. 1: 119-128. krutkiewicz a. 2010. czynniki chorobotwórcze candida albicans. mikol. pol. 17 (2): 134–137. kursat m., erecevit p. 2008. the antimicrobial activities of methanolic extracts of some lamiaceae members collected from turkey. turk. j. sci. technol. 4 (1): 81–85. lópez p., sánchez c., batlle r., nerín c. 2007. vapor-phase activities of cinnamon, thyme, and oregano essential oils and key constituents against foodborne microorganisms. j. agric. food. chem. 55: 4348–4356. manohar v., ingram c., gray j., talpur n. a., echard b. w., bagchi d., preuss h.g. 2001. antifungal activities of origanum oil against candida albicans. mol. cell. biochem. 228: 111–117. pattnaik s., subramanyam v. r., bapaji m., kole c. r. 1997. antibacterial and antifungal activity of aromatic constituents of essential oils. int. microbiol. 89: 39–46. petrie a., sabin c. 2009. medical statistics at a glance, 3rd edition. wiley-blackwell. rex j. h. 1995. serious candida infections: risk factors, treatment and prevention. epidemiology of serious candida infections. pfizer inc., houston. 34 a. głowacka et al. ocena in vitro aktywności przeciwgrzybiczej preparatu „fin candimis” wobec szczepów grzybów z rodzaju candida streszczenie celem pracy była ocena aktywności przeciwgrzybiczej preparatu „fin candimis” (olejku eterycznego oregano) wobec szczepów grzybów z rodzaju candida. do badań wykorzystano 9 szczepów candida albicans oraz 10 szczepów candida glabrata, które wyizolowano z różnych materiałów klinicznych, i 1 szczep wzorcowy candida albicans atcc 90028 oraz preparat „fin candimis”, zawierający olejek eteryczny otrzymany ze świeżych liści oregano. według danych producenta, stężenie czystego olejku eterycznego oregano w produkcie „fin candimis” wynosi 210 mg/ml. wrażliwość szczepów z rodzaju candida na preparat „fin candimis” oznaczano metodą dyfuzyjno-krążkową na podłożu sabouraud’a (po 24-godzinnej inkubacji hodowli w temperaturze 37°c); olejek eteryczny oregano rozcieńczano w 1 ml dmso według postępu geometrycznego. miarą przeciwgrzybiczego działania preparatu „fin candimis” było najmniejsze stężenie hamujące wzrost grzyba (mic). preparat „fin candimis” (olejek eteryczny oregano) może mieć zastosowanie w profilaktyce i leczeniu kandydoz – samodzielnie lub jako naturalny środek wspomagający. szczepy candida albicans są bardziej wrażliwe niż szczepy candida glabrata na preparat „fin candimis”. 2014-01-02t12:00:17+0100 polish botanical society new record of phytophthora root and stem rot of lavendula angustifolia leszek b.orlikowski1 and alma valjuskaite2 1research institute of pomology and floriculture pomologiczna 18, pl-96-100 skierniewice, lorlikow@insad.pl 2lithuanian institute of horticulture, plant protection laboratory babtai, kauno 30, lt-54333 o r l i k o w s k i l.b., va l j u s k a i t e a.: new record of phytophthora root and stem rot of lavendula angustifolia. acta mycol. 42 (2):193-198, 2007. phytophthora cinnamomi was isolated from rotted root and stem parts of lavender as well as from soil taken from containers with diseased plants. additionally botrytis cinerea, fusarium spp. and sclerotinia sclerotiorum were often isolated from diseased tissues. p. cinnamomi colonised leaves and stem parts of 4 lavender species in laboratory trials and caused stem rot of plants in greenhouse experiments. cardinal temperature for in vitro growth were about 7,5 and 32°c with optimum 25-27,5°c. the species colonised stem tissues at temperature ranged from 10° to 32°c. key words: stem rot, isolation, phytophthora cinnamomi, colonisation, temperature, growth introduction lavender (lavendula angustifolia mill., syn. l.officinalis chaix) is mainly medicinal and pharmaceutic plant but since the last years it has been grown in ornamental nurseries mainly as small, blooming, hedge culture. the quality of plants may be strongly decreased by phytophthora species. from diseased plants mainly phytophthora nicotianae var. parasitica was isolated (p a p p a s 1978; p u t m a n 1991; m i n u t o et al. 2001a). growing of plants in italy under shade and in larger pots reduced disease incidence caused by that species (m i n u t o et al. 2001a). variable results were obtained with susceptibility of cultivars or local selections toward the pathogen (m i n u t o et al. 2001b). p a e z et al. (1993) recovered p. palmivora butler from diseased lavendula dentata grown in spanish gardens whereas d a v i n o et al. (2002) isolated the species from rotted root of l. angustifolia in italy. in 2003 on lavender mother plants grown on greenhouse beds yellowing and browning of single shoots were observed. the disease spread slowly on neighbouring acta mycologica vol. 42 (2): 193-198 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 194 l.b. orlikowski and a. valjuskaite shoots. after the next 6 weeks 3/4 parts of affected plants were destroyed (fig.1). on mother plants dark-gray spots on the shoot bases, browning during the next few days and spreading on roots, upwards and on the periphery of affected stems were observed. the disease spread quickly on neighbouring mother plants and cuttings. on plants growing in container grown nursery brown or dark-brown spots developed on individual shoots spread on roots and upwards. during the next 10 days single or a few yellow-brown to dark-brown shoots on individual plants were seen. the development of symptoms were observed during 2 years before or during flowering of plants. the objective of this investigation was to determine a causal agent of lavender shoot and root rot, development of disease in laboratory and greenhouse trials and relationship between temperature and growth of phytophthora cinnamomi and colonisation of stem parts of plants. materials and methods survey of lavender in greenhouse and container grown nursery. on plants grown in greenhouse shoot rot symptoms were observed only in one year on about 5% of lavender. in container grown nursery lavender were grown at least 10 years but shoot and root rot symptoms occurred in years 2004 and 2005 on about 10 and 20%,respectively. diseased plants were collected in plastic bags and transferred to laboratory. the next day plants were removed from pots, substratum were collected in bags whereas plants were washed under tap water and affected shoots were separated, rinsed 3times in distilled water, blotting dried and chosen parts were sterilised over a burner flame. about 5 mm long stem parts were put on pda medium and plates were checked during 4-day-incubation at 24°c in the dark on the presence of phytophthora and other genera. additionally, substratum samples, taken from pots with lavender showing disease symptoms in greenhouse and nursery, were analysed on the presence of phytophthora spp. in years 2003-2005. substratum taken from 3-6 pots was mixed about 5 min in plastic bag and half liter of it was put into plastic box, flooded with tap water (about 1 l) and rhododendron leaves cv. zembla were floated over flooded sample (16 leaf blades/box). boxes covered with plastic foil were incubated at temperature 22-24°c. after 4-6-day-incubation leaves with necrotic spots were removed, washed with distilled water, blotted dried, disinfected over a burner flame and about 5 mm diameter plugs were transferred on pda medium. small parts of colonies, grown around tissues were transferred to pda slants. cultures from diseased stems and substratum were grouped on the base of their growth and morphology and chosen, representative isolates were identified to genera and species (s z k u t a 2004). confirmation of phytophthora classification to species was performed by dna analyses using the method described by o r l i k o w s k i et al. (2006). colonisation of lavender by phytophthora cinnamomi. in laboratory trials leaves and stem parts of 4 cultivars of lavender (tab. 1) were put into plastic boxes on sterilised, wet blotting paper covered with plastic net and inoculated with 3mm plugs taken from the edge of 5-day-old cultures of p. cinnamomi (2 isolates from diseased stem base and substratum) growing on pda at 24°c in the dark. boxes were covered with foil and incubated at 22-24°c in the dark. after 5-day-incubation diameter and length of necrosis was measured. in greenhouse trial stem bases of 3 cultivars of lavender were inoculated with 3 mm diam plugs of p. cinnamomi (from diseased stem new record of phytophthora 195 base) and incubated under covering 12 days. length of necrosis was measured 8 and 12 days after inoculation. relationship between temperature, ranged from 7,5 to 35°c, growth of p. cinnamomi and colonisation of lavender stem parts by the species were studied using the same procedure like in laboratory trials. experimental design was completely randomised with 4 replications and 10 leaves, stem parts or plants in each rep. growth of the species on pda was estimated on the base of 5 petri dishes for each temperature. trials were repeated at least twice. results and discussion isolation of fungi and algae-like oomycetes. similar genera and species were isolated from plants taken from greenhouse and field production (tab. 2). from potential ta b l e 1 phytophthora cinnamomi isolated from substratum used for lavender growth; mean number of dark-brown spots on rhododendron leaves used as the bait source of substratum year of analysis 2003 2004 2005 greenhouse 25.5 b container nursery 14.0 a 10.8 a 17.6 ab note: means followed by the same letter do not differ with 5% of significance (duncan’s multiple range test) ta b l e 2 fungi and algae-like oomycetes isolated from diseased base of lavendula angustifolia; number of settled plants (a) and number of isolates obtained (b) genera/species year of isolation 2003 greenhouse (27 plants) 2004 field (14 plants) 2005 field (21 plants) a b a b a b alternaria alternata nees 2 5 6 11 8 13 botrytis cinerea pers. 7 11 4 9 5 11 cladosporium herbarum (pers.) link. 3 3 2 4 fusarium culmorum (w.g.sm.) sacc. 4 7 2 5 fusarium equiseti (cda) sacc. 3 5 3 7 fusarium solani (mart.) sny. et hans. 6 7 3 5 1 3 humicola grisea traaen 1 2 2 5 mucor spp. 4 8 5 9 7 16 penicillium spp. 3 5 2 4 phytophthora cinnamomi rands 18 56 9 29 18 44 rhizopus sp. 2 4 sclerotinia sclerotiorum (lib.) de bary 4 10 2 7 trichoderma spp. 3 6 3 7 4 7 196 l.b. orlikowski and a. valjuskaite pathogens of lavender botrytis cinerea (mainly in field), two fusarium species, phytophthora cinnamomi and sclerotinia sclerotiorum were detected from diseased plant parts. p. cinnamomi (fig.2) was the most often isolated species both, in greenhouse and field grown plants. the species was detected from 3/4, 5/7 and 6/7 of analysed plants, respectively (tab. 2). isolation of phytophthora cinnamomi from substratum. using rhododendron leaves as the bait, p. cinnamomi was detected from substratum samples taken from 60 pots with plants showing phytophthora rot symptoms. significantly more necrotic spots were observed on rhododendron leaves floated in suspension of substratum collected from diseased lavender grown in greenhouse (tab. 1). colonisation of lavender by phytophthora cinnamomi. both isolates of p. cinnamomi used for lavender inoculation in laboratory trials, caused necrosis of leaves and stem parts of 4 tested cultivars (tab. 3). on stem parts necrosis spread faster than on leaves. in case of leaf blades inoculation, isolate from substratum colonised tissue slower than that from the stem base. on leaves of cv. grosso necrosis spread at least 1/3 faster than on 3 other cultivars. on stem parts such differences were not observed (tab. 3). ta b l e 3 colonisation of leaves and stem parts of lavender by phytophthora cinnamomi; diam/length (in mm) of necrosis after 5-day-incubation inoculation: 2005.09.27 source of p. cinnamomi cultivars leaves stem parts base of stem blue dafo 32.3 ab 43.3 a edelweiss 35.0 bc 40.3 a grosso 32.0 a 39.5 a munstead 35.5 c 38.8 a substratum blue dafo 19.5 b 46.3 b edelweiss 15.3 a 38.8 a grosso 30.8 c 45.3 ab munstead 22.5 b 38.3 a note: means in columns followed by the same letter do not differ with 5% of significance; means separation for the source of the species ta b l e 4 spread of necrosis (in mm) on lavender stems inoculated with phytophthora cinnamomi from the stem base in relation to cultivars and incubation time; greenhouse trials inoculation: 2005.11 17 cultivars length of necrosis in mm after days of incubation 8 12 blue dafo 66.5 b 87.6 b grosso 53.4 a 73.7 ab munstead 48.7 a 65.3 a note: see table 3 198 l.b. orlikowski and a. valjuskaite summary the objective of this investigation was to determine a causal agent of lavender shoot and root rot, development of disease on plant parts and relationship between temperature and growth of pathogen and disease development. during 3 years mycological analyse of diseased lavender, taken from greenhouse and container nursery, was done. additionally, the presence of p. cinnamomi in substratum was estimated using rhododendron leaves as the bait. phytophthora cinnamomi was isolated from the most of analysed plants and from substratum. botrytis cinerea, fusarium spp. and sclerotinia sclerotiorum as other potential pathogens, were recovered from diseased shoot parts. isolates of p. cinnamomi from plant and substratum colonised lavender leaves and stem parts of all tested cultivars. in greenhouse experiment the species quickly colonised stem tissues and necrosis spread about 5,6 mm/24 hr. the growth of the pathogen on pda and colonisation of stem parts were observed at temperature range from about 7,5 to 32,5 °c with optimum 25 -27,5°c. references d a v i n o s., c a c c i o l a s. o., p e n n i s i a. m., d e s t r i n i c o s i a m. g. 2002. phytophthora palmivora a new pathogen of lavender in italy. plant disease 86 (5): 561. m i n u t o a., g u l l i n o m. l., t i t o n e p., g a r i b a l d i a. 2001. influence of cultural practices on incidence of phytophthora nicotianae var. parasitica causing root rot of lavender (lavendula officinalis l.). phytopathol. meditterranea 40 (1): 45–54. m i n u t o g., a r m a t o b., g i l a r d i g., g a r i b a l d i a. 2001. first observations on susceptibility of lavendula spp. to phytophthora nicotianae var. parasitica. informatore fitopatologico 51 (5):69–72. o r l i k o w s k i l.b., tr z e w i k a., w i e j a c h a k. 2006. phytophthora tropicalis, a new pathogen of ornamental plants in poland. j. plant prot. res. (in print). p a e z j. i., b e r r a d., ve g a j. m., te l l o j. 1993. identification de phytophthora palmivora butler en jardines de la exposicion universal de sevilla. bol. de sanidaad vegetal, plagas 19 (4): 633–447. p a p p a s a. 1978. new diseases caused by phytophthora spp. phytophthora newsl. 6: 72–73. p u t n a m m. 1991. root rot of lavender caused by phytophthora nicotianae. plant pathol. 40: 480–482. występowanie zgnilizny korzeni i pędów, powodowanej przez phytophthora cinnamomi, na lavendula angustifolia s t r e s z c z e n i e celem badań było określenie przyczyny zgnilizny pędów i korzeni lawendy, rozwoju choroby w doświadczeniach laboratoryjnych i szklarniowych oraz współzależności pomiędzy temperaturą a wzrostem in vitro patogena oraz kolonizacją części łodyg. w ciągu 3 lat prowadzono analizę mikologiczną chorych roślin lawendy, pobieranych ze szklarni oraz ze szkółki pojemnikowej. dodatkowo, stosując pułapki z liści różanecznika izolowano phytophthora spp. z podłoża. z większości chorych roślin, a także z podłoża izolowano phytophthora cinnamomi. z innych organizmów izolowano botrytis cinerea, fusarium spp. i sclerotinia sclerotiorum. izolaty p. cinnamomi z porażonej rośliny i podłoża kolonizowały liście i części łodyg wszystkich badanych odmian lawendy. w doświadczeniu szklarniowym omawiany gatunek szybko kolonizował tkanki pędów, a nekroza rozwijała się ok. 5,6 mm/ dobę. wzrost p. cinnamomi na pożywce pda oraz kolonizację pędów obserwowano w temperaturze od 7,5° do 32,5°c z optimum 25-27,5°c. fig. 2. zoosporangia of phytophthora cinnamomi and released zoospores. phot. g. szkuta. fig.1. phytophthora shoot and stem rot of lavender cuttings. phot. l. b. orlikowski. 2014-01-01t11:46:10+0100 polish botanical society threats and state of conservation of aphyllophoroid fungi in the mediterranean sergio p. gorjón1 and annarosa bernicchia2 1c/ castaños 16, e-37184 salamanca, spgorjon@gmail.com 2dipartimento di scienze agrarie, università degli studi di bologna, via fanin 42, i-40127 bologna annarosa.bernicchia@unibo.it gorjón s.p., bernicchia a.: threats and state of conservation of aphyllophoroid fungi in the mediterranean. acta mycol. 48 (2): 247–255, 2013. aphyllophoroid fungi is an artificial and diverse group of fungi, often little considered in mycological inventories, but very important in the forest dynamics. in this paper we summarize generally some of the most relevant and interesting habitats in the mediterranean for this group of fungi, and briefly analyze the conservation problematics. key words: corticioids, diversity, europe, polypores, virgin forest, wood inhabiting fungi introduction fungal red-list analysis rarely is considered in national conservation strategies and animals and plants are those who take leadership when delimiting areas and in conservation programs. fungi are only recently starting to be contemplated under a perspective of ecological significance, and most european countries have recently started to produce fungal red-lists. actually, around 5500 different macrofungi are tentatively red-listed in europe (senn-irlet et al. 2007), but there are still large differences in the treatment between different parts of europe. in this respect, there are considerable differences between northern and southern europe and while in some countries fungi are actively considered in conservation programs, in other there are still no official lists and not even diversity and distribution is known for many groups of fungi. the international union for conservation of nature (iucn) is also unequally considering the different groups of living organisms. only three fungal species are listed in two categories (of a total of seven categories) of the 2012 iucn red list (http://www.iucnredlist.org): cladonia perforata a. evans (endangered); erioderma pedicellatum (hue) p.m. jørg. and pleurotus nebrodensis (inzenga) quél. (critically endangered). to compare these two categories with other organisms, in the iucn acta mycologica vol. 48 (2): 247–255 2013 doi: 10.5586/am.2013.026 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 248 s.p. gorjón and a. bernicchia endangered category are listed 3262 species of animals and 2655 species of plants, and under critically endangered are considered 2261 and 1821, respectively. regarding aphyllophoroid fungi, an artificial category represented by a variable mushroom forming fungi, including corticioids, polypores, hydnums, jelly and coralloid fungi, etc., while in some northern european countries (e.g., norway, sweden, and finland) this group has extensively been studied and is actually considered of great importance in connection with the ecology and forest dynamics, in many of the countries from the mediterranean area have even no an accessible checklist or a total or partial ignorance the of diversity. in many parts of southern europe, studies and prospection is still inadequate, and most of the information is dispersed in scientific publications, many of which are difficult to obtain from non specialized people (bernicchia 1999). these differences in the knowledge of the diversity and ecology of this group of fungi between northern and southern europe, are conditioned by several factors such as scientific and mycological tradition, number of professional and amateur mycologists, fundings, and education and environmental awareness, among others. the aim of this work is to reflect on the conservation status and threats of the aphyllophoroid fungi, but is valid by extension to other groups of fungi, and provide some ideas for their preservation, dedicating the present contribution to maria lawrynowicz, in recognition of her work in fungal conservation and in commemoration of the jubilee of her scientific work. mediterranean habitats of interest the mediterranean basin is one of the world biodiversity hotspots (conservation international 2007) and the ecosystem diversity is considerable. hardwood forests of deciduous, marcescent, and perennial species cover the majority of the area, intermixed with coniferous and abundant riparian formations. is not our intention summarize here all the mediterranean ecosystem diversity, but we want to highlight some very interesting forest formations from our mycological experience. coniferous forests the main and most abundant coniferous forests in the mediterranean are those in which different species of pinus l. are predominant. many pine formations are reforestations and, although most of them are naturalized, the diversity and abundance of fungi is quite similar to other parts of europe (some references to this substrate can be found in manjón et al. 1983; bernicchia 2005; bernicchia et al. 2007; bernicchia, gorjón 2010). because of this, we highlight here another most rare and reduced coniferous formations we estimate of great value for the aphyllophoroid diverstity. juniper forests is under our personal appreciation but also because of the associated mycobiota, one of the most interesting mycological area in the mediterranean. juniperus l. is very often a selective substrate for wood-inhabiting rare species as for example antrodia juniperina (murrill) niemelä & ryvarden, arrasia rostrata bernicchia, gorjón & nakasone, echinodontium ryvardenii bernicchia & piga, hyphoderma etruriae bernicchia, lenzitopsis oxycedri malençon & bertault, neolentiporus threats and state of conservation of aphyllophoroid fungi 249 squamosellus (bernicchia & ryvarden) bernicchia & ryvarden, phellinus juniperinus bernicchia & curreli, piloporia sajanensis (parmasto) niemelä, trametes junipericola manjón, moreno & ryvarden, vararia maremmana bernicchia, etc. juniper trees become an irreplaceable substratum, and for this reason the survival of many wood-inhabiting aphyllophoraceous species becomes uncertain. many polypores and corticioids, restricted to very old specimens of juniperus, follow the host genus wherever it occurs, with a scattered distributional pattern. juniper forests are rare and restricted to special areas, presently in decline by natural competition with other species as well as human activity. wood of several species of juniper has traditionally been used to build houses, fences, and huts. although its use is currently limited and replaced by other materials, it is prioritary to promote its conservation. the reader is referred to the the studies focused on the diversity of aphyllophoroid fungi on juniperus (manjón, moreno 1981; bernicchia 2000). another endangered coniferous formation is those of abies pinsapo boiss., actually restricted to southern spain and northern morocco. a very interesting catalogue of wood-inhabiting fungi was reported by manjón and moreno (1983b). the conservation of this relict area is evidently priority. some other conifers should be better considered by its rarity or as potentially interesting substrate. few mycological studies have been made in abies alba mill. (bernicchia et al. 2007b) and larix decidua mill. a remarkable conifer is cedrus atlantica (endl.) manetti ex carrière, where only few species have been reported and three corticioid fungi are only know from this substrate: aleurodiscus atlanticus maire, neoaleurodiscus monilifer (malençon) sheng h. wu, and the recently described acanthophysellum verecundum duhem (duhem 2001). hardwood forests in the mediterranean area the main hardwood forests are formed by several oak species, forming monospecific or more commonly mixed forests with several shrubs. each species tends to have its own associated fungi, and some aphyllophoroid species show an exclusively mediterranean distribution. sarcodon quercinofibulatus pérez-degreg., macau & j. carbó has recently been described and reported growing associates mainly to fagaceae in spain and italy (vizzini et al. 2013). ramaria mediterranea schild & franchi, has exclusively been found growing in the mediterranean area in mixed forest related to various quercus l. species and other mediterraneous shrubs (däniels, gorjón 2009). some species of corticioid fungi and polypores follow the quercus distribution, even they also can be sporadically growth on other substrata, as for example, aleurocystidiellum disciforme (dc.) tellería, hexagonia nitida durieu & mont., marchandiopsis quercina (j. erikss. & ryvarden) ghobad-nejhad, peniophora quercina (pers.) cooke, and piptoporus quercinus (schrad.) p. karst. for further information on aphyllophoroid fungi growing on quercus see bernicchia et al. (2008). mediterranean shrublands some species are linked to mediterranean shrubs. a conspicuous polypore, antrodia sandaliae bernicchia & ryvarden, has exclusively been found in italy and spain growing specifically on arbutus unedo l. (gorjón, bernicchia 2009). that substratum proved particularly interesting for the numerous aphyllophoroid species growing on 250 s.p. gorjón and a. bernicchia it (gorjón et al. 2006). also, there are a large number of corticioid fungi inhabit-gorjón et al. 2006). also, there are a large number of corticioid fungi inhabit-also, there are a large number of corticioid fungi inhabiting leaves, small twigs, and stems associated to mediterranean shrubs. recently, aphanobasidium gloeocystidiatum duhem and vuilleminia ericae duhem have been described growing on cistus monspeliensis l. and erica arborea l., respectively (duhem 2010). usually, conservation has focused their efforts in protect large forested areas but ecosystems as the mediterranean marquis shrubland and garrigue, predominantly formed by varied small shrubs, are also interesting to be considered in an integral protection of the mediterranean biodiversity. riparian forest generally, species of alnus mill., betula l., celtis l , corylus l., fraxinus l., populus l , salix l., and ulmus l , and are associated to permanent watercourses, forming the majority of the riparian forests in the mediterranean. seasonal or intermittent flow courses are also very common in the mediterranean region having a vegetation of its own, dominated by nerium oleander l., tamarix gallica l., and securinega tinctoria (l.) rothm. some species of aphyllophoroid fungi associated to riparian trees are in example, phanerochaete tamariciphila boidin, lanq. & gilles, trametes suaveolens (l.) fr., and vuilleminia coryli boidin, lanq. & gilles. because of its rarity and restricted distribution, some of them are priority to be conserved, and many of these forests are critically endangered by certain human activities and most severely by the construction of hydroelectric stations and big dams. by the abundance of litter and dead wood in this ecosystems there are a number of wood-decaying species of polypores and corticioids. extreme or particular habitats and hosts in the mediterranean area some particular habitats that have a restricted distribution, relicts of wider forests of the past, are very interesting and peculiar from a mycological point of view, with some threatened wood-inhabiting fungi associated. one of these is the ilex aquifolium l. and taxus baccata l. forest, a special and with a very restricted distributional area where the common holly grows in association with pluricentenarian european yews. those taxus baccata trees are considered among the oldest giants in europe. there are some good formations in the northern of the iberian peninsula and in the northern part of sardinia. in these last italian forests, very peculiar indeed, has exclusively been collected aleurodiscus ilexicola bernicchia & ryvarden, while dendrothele tetracornis boidin & duhem, and d. wojewodae pouzar can be considered very rare species with a restricted distribution in europe. another example is the dunal habitat. it is a particular, unstable, almost lost and very interesting ambient, that shows extreme growing conditions. it is present sporadically here and there where juniperus, pistacia l., erica l., phillyrea l., etc., colonize the sands of the dunes. it is one of the most interesting habitat for woodinhabiting aphyllophoraceous species even if the growing period is very short, due to the windy conditions, the salinity, and its connection to the necessity of high humidity levels. the number of species is never numerous but very often they are unique like acanthophysellum dextrinoideocerussatum (manjón, m.n. blanco & g. moreno) sheng h. wu, boidin & c.y. chien, phellinus pseudopunctatus a. david, dequatre & fiasson, and p. rosmarini bernicchia, and vararia maremmana. threats and state of conservation of aphyllophoroid fungi 251 european red list of fungi there is no an official european red list of fungi, and progress to produce a checklist of endangered fungi is different among the european countries. the european council for the conservation of fungi (eccf) has compiled a preliminar european red list of endangered macrofungi (http://www.wsl.ch/eccf/candlist-subtotals. xls) where list a total of 1643 candidates distributed in the next orders: agaricales (794 especies), atheliales (1), boletales (45), cantharellales (13), ceratobasidiales (1), corticiales (6), dacrymycetales (2), elaphomycetales (13), geastrales (4), gomphales (9), helotiales (72), hymenochaetales (32), hypocreales (14), pezizales (202), phallales (65), polyporales (214), russulales (95), trechisporales (3), thelephorales (32), tremellales (6), and xylariales (20). aphyllophoroid fungi are distributed among all the major orders of basidiomycetes, and almost 400 species are considered in the previous checklist. however, the list has some shortcomings and they are missing newly described species, but it serves as base list of species, and consequently habitats, to be preserved. problematic and threats there are several publications about fungal conservation and the reader can find more information and other reading suggestions in moore et al. (2001). we will briefly and generally review below some of the problems and possible solutions to achieve an adequate conservation of habitats and species of fungi. unequal treatment of different groups of fungi traditionally, some groups as agarics or bolets have received a considerably broad consideration by both, professional and amateur mycologists. contrary, some groups and genera of basidiomycetes and ascomycetes are neglected in most inventory studies. mycologists, above all, non professional mycologists, are less interested in some groups visual less attractive and where the need of a microscope is of critical importance and a limitation, resulting in the loss of interest for them. also, some groups, with non pathogenic or agricultural importance, are usually discriminated by the administrations and to get funds for their study is much more difficult. inadequate knowledge of the diversity one problem to remark is the poor knowledge of some groups of fungi, and for the aphyllophoroid fungi this is very patent in several families. for many species there are only few records (also is usual that only the type specimen is known and never recollected) and the taxonomical, ecological, and chorological information is very restricted. to add more confusion, species delimitation is still incomplete in many species complexes. molecular information, when available, is often of exceptional value to resolve phylogenetical relationships, but also in some cases it contributes to add some confusion when molecular analysis is incomplete or deficient. europe is probably the better explored and studied area in the world, but several new species 252 s.p. gorjón and a. bernicchia are still described every year. to illustrate this, two new corticioid genera where recently described (bernicchia et al. 2011, duhem & schultheis 2011), but also more conspicuous species are described each year as in the case of two new species of hydnum l. (olariaga et al. 2012) and a sarcodon quél. ex p. karst. (pérez-de-gregorio et al. 2011). also, differences between countries or regions are patent and inside europe, while in portugal, spain, france, and italy, the knowledge is quite acceptable, in eastern countries there are several deficiencies. the situation is even worst in north african countries, and the research advance is here fundamental to have a complete understanding of the mediterranean area. habitat fragmentation and reduction this is probably the main problem regarding world conservation of the biodiversity. considering the mediterranean area a very populated place, it is a difficult challenge to make compatible the protection of the ecosystems and the demographic expansion, and the recently abusive and uncontrolled tourism style has not helped. most of the forested areas have been reduced or eliminated and replaced by farming lands, roads, urbanized areas, and large tourist resorts for example, with an abusive use of resources and land colonization. in addition, in the last decades, large areas of native forests have been replaced by pine, poplar, and eucalyptus cultivations. also, one of the biggest problems in the mediterranean area for the conservation of forests is the risk of fire. we must not forget nevertheless that fire in the mediterranean area is a natural method of regeneration conditioned by natural factors that would not cause major problems. unfortunately, fires from natural causes are limited and more than 95% of fires in mediterranean areas are intentional or caused by negligences, constituting a serious risk for the preservation of the biodiversity. desertification some areas in the mediterranean have experimented serious problems of desertification in the last decades. the problem is complex but it is imperative to do a responsible water and forest management promoting reforestations programs. in this sense the irresponsible housing bubble of the last years on the mediterranean, has greatly contributed to the degradation of various forest environments and the situation is difficult to reverse in many of them. poor coordination between administration, research institutions and inadequate funds in the majority of cases, research is unfortunately not coordinate with the public administration. often, results and advances of the research or individual projects are not further considered in public management and important information is restricted to academic or scientific circles and mycological journals. scientific progress is not immediately applied in forest management or environmental legislation with the consequent lost of utility. basic research, biodiversity, and ecological studies with no important economic repercussion are usually considered less relevant and relegate to a second place. we must not forget the importance of basic science for the progress of other researches, threats and state of conservation of aphyllophoroid fungi 253 and in this time of reduction of the biodiversity is essential to concentrate efforts in the knowledge and the protection of the natural resources. in the present time, when in general, the national governments do severe cuts to research, is also necessary to get support and funding in the private sector, such as foundations or organizations interested in conservation and environmental education. strategies of conservation highlight fungal importance fungi play an important role in various ecosystems. wood rotting fungi, mycorhizogenous, and parasitic species are essentials in the forest dynamics. nutrient cycle, soil structure, and forest productivity is largely regulated by fungi. also, there is an underexplored field regarding chemical compounds and several species may be investigated as potentially important in medicine, natural therapy, nutrition, or organic dyers, as example. increase research in poor or unexplored areas as the knowledge of the aphyllophoroid biodiversity is still incomplete, it is mandatory to increase inventory work, in order to find out what and why we need to conserve and protect. it would be adequate that the research results were reflected not only in scientific journals, usually inaccessible to non specialized people, and try to disseminate as much as possible the acquired knowledge. protect key and vulnerable species we need to be more incisive and try to convince the administration for a rapid inclusion of the most vulnerable species in the national and european legislation. sometimes it is difficult to approach the bureaucratic and legislative steps, but is much simpler try to sensitize forest managers and local people of the need to include fungi in the forest management planning. in this sense is essential to have good communication with directors, park rangers, and managers of the natural areas that are generally very receptive to the information exchange. promote conservation of priority habitats and creation of microreserves in coordination with botanists and zoologists, we need to emphasize the protection of the habitats of the directive 92/43/eec. under the guidance of the previous directive, large natural areas in europe have been mapped, but just under a strictly botanical perspective. also several microreserves (singular, small areas of no more than 20 hectares) have recently been created for the protection of rare, endemic, or characteristic floristic elements. it would be interesting and necessary to combine the economic effort of the public administrations to get an integration of all biotic elements when proposing such reserves, and consideration of fungi is essential, as significant elements in the forest ecology. 254 s.p. gorjón and a. bernicchia good forest and agricultural practices forest management and the proper use of agricultural and pastoral areas are essential to the conservation of biodiversity. traditionally, the administration of forested areas in much of the mediterranean has been led out by forest engineers, and unfortunately in many cases from the point of view of timber production. fortunately today, multidiciplinares teams handle their management and forests are beginning to be considered not only from a purely productive perspective but as a source of varied resources and many areas are being recolonized by original forests. the maintenance of different successional stages, adequate management favoring the abundance and decomposition of organic debris, and an equilibrate forestry are compatible to maintain a high fungal diversity. the change from extensive to intensive land use, has also enabled the regeneration of large areas. however, traditional farming is clearly possible and recommendable, following a few good practices compatibilizing land use and the presence of forests. environmental education this is probably the most important step in the immediate conservation of biodiversity. educators have an essential role to play but it is also indispensable the efforts of individuals, mycological associations, foundations, 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(eds). 2001. fungal conservation: issues and solutions. cambridge university press. 272 pp. olariaga i., grebenc t., salcedo i., martín m.p. 2012. two new species of hydnum with ovoid basidiospores: h. ovoideisporum and h. vesterholtii. mycologia 104: 1443-1455. pérez-de-gregorio m.à., macau n., carbó j. 2011. sarcodon quercinofibulatum, una nueva especie del género con hifas fibulíferas. rev. cat. micol. 33: 25-30. senn-irlet b., heilmann-clausen j., genney d., dahlberg a. 2007. guidance for conservation of macrofungi in europe. document prepared for the directorate of culture and cultural and natural heritage, council of europe, strasbourg. (http://www.wsl.ch/eccf/guidance_fungi.pdf) vizzini a., carbone m., boccardo f., ercole e. 2013. molecular validation of sarcodon quercinofibulatus, a species of the s. imbricatus complex associated with fagaceae, and notes on sarcodon. mycol. progress (on-line first). 2013-12-20t14:49:35+0100 polish botanical society soil fungi communities from young scots pine plantations affected with root rot wojciech szewczyk department of forest pathology, agricultural university wojska polskiego 71c, pl-60-625 poznań, wszew@au.poznan.pl s z e w c z y k w.: soil fungi communities from young scots pine plantations affected with root rot. acta mycol. 42 (2):239-244, 2007. the aim of work was to investigate the qualitative and quantitative characteristic features of soil fungi associations occurring in i class pine stands threatened by pathogens of tree roots in zielonka experimental forest district (w poland). during the mycological analyses of soil, a total of 694 isolates of fungi representing 33 species were obtained. the most numerously represented species was penicillium daleae which made 75% of all isolates. the second place in the ranking belonged to penicillium janczewski amounting to 11%. in order to define the qualitative and quantitative similarity between the studies areas, a proportional inconformity was determined. the greatest similarity occurred between the associations of soil fungi obtained from site 64c and 35b. key words: soil fungi, scots pine, plantations introduction fungi occur in all environments; they play an important role, both a positive and negative one, in the life of other organisms and ecosystems. soil environment is very favorable for fungi, they participate in the decomposition of dead organic matter, particularly in conditions of a low ph level which limits the activity of bacteria, and they enter into mycorhizal relations with trees. there is no doubt that there exist many factors affecting the quality of relations dominating in the soil determining thereby the occurrence of the particular species of organisms or whole organism associations. associations of organisms occurring on a given area together with arbores cent species decide about the quality of ecological factors in the given place reacting to all changes taking place there (m a ń k a 1974). therefore, it is important to get familiar with the biodiversity of these associations (k o w a l s k i 1996) which can decide about the health of plants (d a h m , s t r z e l c z y k 1977). the objective of the present study was to investigate the qualitative and quantitative characteristic acta mycologica vol. 42 (2): 239-244 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 240 w. szewczyk features of soil fungi associations occurring in i class pine stands threatened by pathogens of tree roots. materials and methods studies were carried aut in seven separated sites (tab. 1) localized in i class pine stands of the experimental forest district zielonka (w poland). in each separated site, soil samples were taken from one studied area in order to isolate fungi communities. soil for mycological analyses was taken from the depth of 5-20 cm from 6 sites uniformly distributed on a circle with an about 6 m radius. the obtained 6 samples were mixed, givining in this way a sample of about 1 dm3 volume. fungi were isolated using warcup’s method (wa r c u p 1950) in the modification of k. mańka (j e n s e n , m a ń k a 1961; m a ń k a , s a l m o n o w i c z 1987). from each flask containing soil and sand, 10 petri dishes were prepared. in this way, the mycological soil analysis from one area was based on 30 petri dishes. incubation was carried out for 7 days at room temperature. after this period, the first colonies of fungi which developed on petri dishes were separated. fungi separation was repeated after 3 successive days. the developed fungi colonies were transferred to test-tubes with agar-glucose-potato nutrients. after 2-3 weeks of incubation, the separated isolates were described. the description included the dish number from which the colony of fungi was separated and the successive number of test-tube. then, the developed mycelia were compared and groups with the same macroscopic features were separated. after 2 weeks, the representatives of these particular groups were transferred to petri dishes with agar-glucose-potato nutrient. after 10 days of incubation, the fungi were identified and described. in this way, soil fungi associations obtained and systematized. ta b l e 1 characteristic and location investigation area compartment forest range forest site type pine age in 2002 47k huta pusta fresh mixed coniferous forest 9 43f kamińsko fresh coniferous forest 12 64c kamińsko fresh mixed broadleaved forest 10 85c potasze fresh mixed coniferous forest 9 35b huta pusta fresh coniferous forest 13 5a rakownia fresh mixed broadleaved forest 13 142d stęszewko fresh mixed coniferous forest 13 soil fungi 241 in order to determine the qualitative and quantitative similarities of the soil fungi communities obtained from the studied areas, on the basis of the number of common and different species, the formula of marczewski-steinhaus (r o m a n i s z y n 1972; l e ś n i a k 1984; s i e r o t a 1995; k o w a l s k i 1996; ty s z k i e w i c z 2001) was applied. s(1,2)= w a+b – w where: s – similarity of two mutually comparable soil fungi communities a – number of isolates of fungi communities 1, b – number of isolates of fungi communities 2, w – number of fungi association isolates common for both associations (1 and 2). it is assumed that if the comparable communities have all features in common, the similarity is s = 1 or 100%. if the studied communities have no common features, then their similarity is s = 0. also the so called distance between the communities was determined: the more features are common, the smaller is the distance, and the less features are common, the greater is the distance. the distance of two associations (r) is expressed by the formula: r = 1 – s because of the discrete character of the data, in order to define the qualitative and quantitative similarity between the studies areas, a proportional inconformity was determined. the distance between the areas has been illustrated on dendrograms using the method of mean connections (d o b o s z 2001). results during the mycological analyses of soil, a total of 694 isolates of fungi representing 33 species were obtained. the most numerously represented species was penicillium daleae which made 75% of all isolates and it was present in all associations from all studied sites. the second place in the ranking belonged to penicillium janczewski amounting to 11% and it was also present in all the studied associations except for the association from site 47k. the successive rank belonged to chrysosporium merdarium (7%), ch. pannorum (7%) and penicillium stecki (4%). species group which occurred sporadically, they were represented by one isolate only, included: mortierella isabelina, m. microspora var. macrocystis and two non-sporiferous species. the most numerous soil fungi association was obtained from site 85c, it consisted of 179 isolates representing 13 species. the least numerous was the community of soil fungi from site 47k including 63 isolates represented by 10 species. the poorest community regarding its qualitative aspect was the association from site 35b represented by 9 species of soil fungi. table 2 contains the qualitative and quantitative composition of soil fungi associations from the particular studied areas. 242 w. szewczyk in order to determine the qualitative similarities of fungi associations, on seven analyzed areas, similarity cocfficients of associations were determined according to the formula of marczewski-steinhaus (tab. 3). similarity determined in this way shows only the fact of occurrence or absence of the given fungi, but it does not indicate their frequency. ta b l e 2 species of fungi obtained on investigation areas species area total5a 35b 43f 47k 64c 85c 142d absidia glauca hagem 2 1 3 acremonium charticola (j. lindau) w. gams 2 2 acremonium killiense grütz 3 1 4 acremonium pteridii w. gams et frankland 2 2 acremonium strictum w. gams 1 2 1 4 alternaria alternata (fr.) keissl. 4 1 5 aspergillus clavatus desm. 4 2 6 chaetomium globosum kunze (ehrenb.) j.w. carmich 3 3 chrysosporium merdarium (ehrenb.) j. w. carmich 1 32 15 48 geomyces pannorum sigleret j. w. carmich 10 9 3 25 47 mortierella isabellina (oudem.) arx 1 1 mortierella macrocystis (w.gams) 1 1 umbelopsis nana (linnem.) arx 2 2 mortierella vinacea dixon-steward 3 2 5 non-sporiferous 1 1 1 non-sporiferous 2 2 2 non-sporiferous 3 8 8 non-sporiferous 4 1 1 penicillium adametzii zaleski 3 5 2 penicillium citrinum thom 9 2 13 9 11 44 penicillium daleae zaleski 52 69 40 46 39 74 36 356 penicillium glabrum (wenmer) westling 6 1 7 penicillium janczewskii zaleski 18 2 14 24 13 5 76 penicillium jensenii zaleski 1 4 5 penicillium minioluteum dierckx 2 2 penicillium vinaceum gilman et abbott 8 8 penicillium waksmanii zaleski 7 1 1 9 trichoderma aureoviride rifai 3 1 5 9 trichoderma koningii oudem. 2 2 trichoderma viride pers. 4 1 7 12 simplicillium lamellicola (f.e.v. sm.) w. gams 3 1 4 micromucor ramannianus var. ramannianus (a. möller) arx 5 5 total: 112 90 79 63 92 179 80 694 ta b l e 3 similarities of soil fungi communities coming from compartment in zielonka experimental forest district (w poland) 142d 85c 64c 47k 43f 35b 5a 28% 14% 20% 24% 17% 18% 35b 24% 29% 60% 27% 19% 43f 22% 21% 21% 11% 47k 16% 10% 31% 64c 19% 25% 85c 19% soil fungi 243 the greatest similarity. as height as 60% occurred in case of soil fungi communities from sites 64c and 35b. the least similarities were found between the communities of soil fungi from sites 47k and 43f amounting only to 11%. other soil fungi associations showed a similar qualitative similarity ranging between 14 and 31%. during the studies, the coefficient of the proportional distance between the associations was determined taking into consideration also the frequency of the given fungi species in the associations. the obtained coefficient values have been illustrated in the form of a dendrogram (fig. 1). the greatest similarity occurred between the communities of soil fungi obtained from site 64c and 35b. the communities of soil fungi obtained from site 85c was the most different from the other ones. discussion on the studies areas where soil samples were taken for mycological analyses, observation were carried out referring to the monitoring of infections diseases of tree roots (s z e w c z y k 2005). on all studied areas, there occurred pine infestations by a. ostoyae. an exception was site 35b where pine were attacked by h. annosum. in all studied areas there grew young planted trees, the soil was prepared in the same way (furrow ploughing) and the trees were in a similar age (9-13 years), however, they occupied different forest sites (fresh coniferous forest, fresh mixed coniferous forest and fresh mixed broadleaved forest). on the basis of a determined similarity of associations and the coefficient of proportional distance, one can conclude fig. 1. percentage distance between soil fungi community coming from compartment in zielonka experimental forest district. 244 w. szewczyk that there is no dependence between the qualitative and quantitative composition of soil fungi and the occurrence of armillaria ostoyae and heterobasidion annsoum, the forest sites type and the age of trees on the areas where samples were taken. all obtained isolates represented commonly occurring soil fungi. some of them can have an effect on armillaria spp. or heterobasidion annosum. the most frequently occurring fungus type was penicillium janczewski characterized by the production of enzymes which decompose cellular walls of wood making thereby an entrance for infection of armillaria spp. (b ä ä t h , s o d e r s t r ö m 1980). another type of fungus which may have an effect of armillaria spp. is trichoderma which trough antibiosis can limit the growth of rhizomorphs (s h o w , k i l e 2001). fungi of mortierella type do not play any significant role in soil metabolism, but they are able to decompose chitin acremonium spp. can be pathogens for humans and animals: the same refers also to aspergillus spp. or penicillium spp. causing sapergollotoxicoses and penicillotoxicoses. references b ä ä t h e., s ö d e r s t r ö m b.e. 1980. degradation of macromolecules by microfungi isolated from different podzolic soil horizons. can. j. bot. 58: 422–425. d o b o s z m. 2001. wspomagana komputerowo statystyczna analiza wyników badań. akademicka oficyna wydawnicza exit, warszawa. j o h n s o n l.f., m a ń k a k. 1961. a modification of warcup’s soil-plate method for isolating soil fungi. soil sci. 92: 79–84. k o w a l s k i s. 1996. biodiversity of soil fungi in converted stand of pinus sylverstris l. as an indicator of environment degradation as the effect of industrial pollution. phytopathol. pol.12: 163–175. m a ń k a k., s a l m o n o w i c z b. 1987. udoskonalenie niektórych technik zmodyfikowanej metody płytek glebowych do izolowania grzybów z gleby z punktu widzenia mikologii fitopatologicznej. rocz. nauk rol. ser. e ochr. rośl. 17: 35–46. r o m a n i s z y n w. 1972. uwagi krytyczne o definicji sorensena i metodzie renkonena obliczania współczynnika podobieństwa zbiorów. wiad. ekol. 18 (4): 375–380. s z e w c z y k w. 2005. monitoring armillaria root rot in young (up to 20 yrs) scots pine plantation in zielonka forest district. acta sci. pol. silv. colendar. rat. ind. lignar. 4 (2): 91–100. ty s z k i e w i c z z. 2001. zbiorowiska grzybów glebowych i ich wpływ na kształtowanie się zdrowotności naturalnych odnowień dwóch różnych grądów. zesz. nauk. politechniki białostockiej. inżynieria środowiska 12. białystok: 143–185. wa r c u p j. 1950. the soil plate method for isolation of fungi from soil. nature 166: 117–118. zbiorowiska grzybów glebowych w młodych plantacjach sosny zwyczajnej atakowanej zgnilizną korzeni s t r e s z c z e n i e celem badań było poznanie i porównanie zbiorowisk grzybów występujących w glebach leśnych wybranych siedlisk na terenie nadleśnictwa doświadczalnego zielonka. w trakcie analiz mikologicznych gleby otrzymano ogółem 694 izolaty grzybów reprezentujące 33 gatunki. najliczniej reprezentowanym gatunkiem był penicillium daleae, który stanowił 75% wszystkich izolatów i wchodził w skład zbiorowisk ze wszystkich badanych wydzieleń. drugim pod względem frekwencji był penicillium janczewskii – jego udział w ogólnej liczbie izolatów wynosił 11 %. wyznaczono współczynnik odległości procentowej między zbiorowiskami. największe podobieństwo występuje między zbiorowiskami grzybów glebowych uzyskanych z powierzchni badawczych zlokalizowanych w wydzieleniu 64c i 35b. 2014-01-01t11:46:31+0100 polish botanical society site characteristics of tuber magnatum in greece vasilios christopoulos¹, polyxeni psoma² and stephanos diamandis¹ ¹hao-forest research institute, el-570 06 vassilika, thessaloniki, diamandi@fri.gr ²hao-soil science institute, el-570 01 thermi, thessaloniki christopoulos v., psoma p., diamandis s.: site characteristics of tuber magnatum in greece. acta mycol. 48 (1): 27–32, 2013. day after day it is being demonstrated that greece hides a buried treasure in its forests. the wild black truffles tuber aestivum, t. uncinatum, t. brumale and t. melanosporum and the white truffle t. borchii and recently t. magnatum are picked in a variety of forest ecosystems all over the country. the information which has been collected has initiated a national programme on truffle cultivation which, so far, has immense appeal, especially among young farmers. the discovery of carpophores of t. magnatum, the most valued white truffle in the market, triggered the interest in studying the site characteristics, botanical and pedological, in order to help farmers to cultivate the species in similar sites. all carpophores were found under hornbeam (carpinus orientalis) along the banks of a creek on alluvial sediment. soil analyses of four soil samples from truffle nests showed a narrow range of values of ph in water (7.57–7.78) and 1n kcl (6.94–7.07) and a similar granulometric soil texture. key words: white truffle, soil analyses, ecology and distribution, truficulture introduction over the last fifteen years there has been considerable hope and speculation that since the forests and soil types in greece are similar to those of neighbouring italy, truffles should exist in greece too. due to a lack of data, however, these hopes could not be proven. the first specimen of an immature carpophore of t. aestivum vittad. (black summer truffle) was officially sent to the forest research institute in 2003. the specimen was identified by dr stephanos diamandis and then confirmed by prof. maria lawrynowicz from lodz university, poland. soon afterward, a series of seminars was initiated by the forest research institute in areas of greece deemed suitable for truffle. this resulted in the emergence of truffle hunters who sent in a great deal of data confirming that the climatic and soil conditions as well as host trees in greece were suitable for truffle growing. acta mycologica vol. 48 (1): 27–32 2013 doi: 10.5586/am.2013.004 28 v. christopoulos et al. black truffles of commercial interest, such as tuber aestivum (summer truffle), t. uncinatum chatin (bourgundy truffle), t. brumale vittad. (winter truffle), t. melanosporum vittad. and the white truffles, t. borchii vittad. (biancetto) and t. excavatum, growing in the wild are now picked in a variety of forest ecosystems all over the country (diamandis and perlerou 2008). a remarkable number of young farmers have responded positively to a national programme encouraging truffle cultivation. all of the species mentioned above are now cultivated in a variety of mountain areas from the northern border all the way down to crete, with the exception of t. magnatum pico which had never been found in greece. recently, however, the forest research institute received its first specimens of t. magnatum (figs 1 and 2), which is widely considered as the most valued but at the same time the most difficult truffle to find and to cultivate. these are the first preliminary results in an attempt to study the site characteristics of t. magnatum truffle-beds in greece in order to encourage its cultivation in similar sites. the authors, respecting the wish of the collectors, do not provide any geographical information about specific locations. materials and methods the site features, botanical and soil physico-chemical characteristics, were studied during a field visit to the one single location where the first white truffles had been collected. soil samples were received from four truffle-nests along a creek where carpophores had been collected. a core of soil 30 cm deep was removed by using a common vertical spade and transferred into plastic bags. four such soil samples were taken 15 cm away from an equal number of truffle-nests and one additional soil sample was taken from the same area but away from truffle-nests to be used as control. the distance between the first and the last truffle-nest was about 300 m. the soil analyses were carried out by using standard laboratory methods. in particular, the soil texture was determined by the hydrometer method (gee, bauder 1986; gee, or 2002). metal concentrations (fe, zn, mn and cu) were determined by using inductively coupled plasma emission spectroscopy (icp) after extraction with dtpa solution at ph 7.3 (lindsay, norvell 1978). the ph was assessed both in a saturation paste and in a solution of 1:1 of 1n kcl. the chemical parameters that were also determined were total caco3 by back titration (piper 1942), organic matter by the wet digestion method (walkley, black’s 1934), phosphorus (p) by the olsen method (olsen et al. 1954), potassium (k), calcium (ca) and magnesium (mg) by the ammonium acetate method (knudsen et al. 1982) and boron (b) by the hot water method (bingham 1982). the results of the analyses are demonstrated in table 1. results the area referred to is only 15 km away from the sea. the climate is mediterranean with warm, wet winters and a long dry season through summer. annual mean temperature in 2012 was 16° c while mean minimum was 10.8° c. total precipitation in 2012 was 775.2 mm. site characteristics of tuber magnatum in greece 29 the carpophores of t. magnatum were collected at the banks of a narrow creek with steep sides and under natural carpinus orientalis. other tree species along the creek were quercus pedunculiformis, salix sp. and corylus avellana while the dominant plant of the undergrowth was ruscus aculeatus. fig. 1. carpophore of tuber magnatum showing irregular, lobed and compressed shape as well as decoration of gleba. fig. 2. tuber magnatum. ascus with three broadly ellipsoid ascospores. 30 v. christopoulos et al. the specific truffle-nests were located 50-80 cm above the water surface on rich soil that had accumulated on the narrow banks by falling from the steep sides. there was an abundance of thin tree roots around the nests. according to the figures reached by granulometry, the soil structure is defined as l to cl. the results of the soil analyses showed that soil around the truffle-nests is alkaline with the ph values ranging between 7.57 and 7.78. ph values in kcl also had a narrow range, 6.94 to 7.07. the ph values of the control sample were not significantly different. the values of all parameters, except those of k and ca content, between the four truffle soil samples and the control soil sample were not significantly different either. k and ca content in the control sample were, however, significantly higher. results of the soil analyses are shown in table 1. table 1 results of soil analyses. samples b1-b4 refer to truffle-nest samples. b5 refers to the control sample sample no. soil struc ture clay % silt % sand % ph in h2o ph in kcl ec ms/ cm caco3 % organic matter % β1 cl 34 32.0 34.0 7.78 7.07 0.89 70 5.81 β2 cl 30 34.0 36.0 7.65 6.94 0.79 0.3 5.24 β3 l 18 44.0 38.0 7.57 7.02 0.48 1.5 2.01 β4 cl 34 32.0 34.0 7.60 7.02 0.84 5.0 6.61 β5 control cl 28 34.0 38.0 7.84 7.25 0.86 20.0 5.46 sample no. no3 ppm p ppm k ppm mg ppm fe ppm zn ppm mn ppm cu ppm b ppm ca ppm b1 23.99 6.92 159 195 26.13 1.11 19.22 1.31 0.73 5960 b2 31.01 2.19 142 181 24.49 1.36 13.30 1.05 0.85 3679 b3 10.92 1.79 69 106 17.54 0.46 7.03 0.62 0.53 3661 b4 46.80 3.24 143 194 29.66 1.70 19.64 1.81 0.73 5730 b5 control 45.83 4.45 268 193 28.49 1.40 21.66 1.48 0.98 7879 table 2 shows the range of values of the physico-chemical variables between the truffle-nests and the control sample which were determined. table 2 comparative range of physico-chemical soil variables between truffle-nest soil and control sample clay % silt % sand % ph in h2o ph in kcl ec ms/cm caco3 % organic matter % trufflenests 18-34 32-44 34-38 7.577.78 6.94-7.07 0.48-0.89 0.3-7.0 2.01-5.81 control 28 34 38 7.84 7.25 0.86 20.0 5.46 sample no3 ppm p ppm k ppm mg ppm fe ppm zn ppm mn ppm cu ppm b ppm ca ppm truffle nests 10.9246.80 1.796.92 69159 106195 17.5429.66 0.461.71 7.0319.64 0.621.81 0.530.85 3661-5960 control 45.83 4.45 268 193 28.49 1.40 21.66 1.48 0.80 7879 site characteristics of tuber magnatum in greece 31 the caco3 content of the control sample appears significantly higher (20.0) than the soil around the truffle-nests (0.3-7.0). significantly higher is also the k and ca content. all five samples which came from the same soil horizon had good porosity and drainage, satisfactory aeration and weakly developed granular structure whereas the clay content ensured persistent minimum moisture. discussion carpophores of tuber magnatum are recorded in the wild for the first time in greece. as interest in truffle cultivation is steadily growing, an effort was undertaken to analyze the soil around the first white truffle-nests in order to be able to promote white truffle cultivation in cases where soils are similar in physical and chemical properties. the present work is a first approach based on a limited number of soil samples taken from truffle-nests along a creek at a single location. therefore, they can be considered only as preliminary results. the particular fungus at the specific location seems to create mycorrhiza with carpinus orientalis. its occurrence on alluvial calcareous deposits comes in agreement with habitats in italy where the fungus is intensely hunted (lulli, primavera 2001; raglione, owczarek 2005). as with t. melanosporum, the physico-chemical characteristics of the soils may show a strong variability within each parameter (bencivenga et al. 1988; poitou 1988; raglione et al. 1992, 2001). in our case the soil analyses showed a narrow variability, especially in the values of ph in water and 1n kcl, as well as in the other chemical parameters determined among the truffle-nests, but this is to be expected since the soil samples came from the same alluvial deposits along the same creek. corresponding values of the control soil sample did not differ significantly from those of the truffle-nests except those of caco3, k and ca content. the limited number of samples, however, does not allow any conclusions to be drawn regarding their significance. acknowledgments. the authors would like to thank ms. e. tziatziou and mr. v. goulandas who found the truffles and helped to collect the soil samples. anonymous reviewers are thanked for valuable suggestions to improve this manuscript. references bencivenga m., calandra r., granetti b. 1988. ricerche sui terreni e sulla flora delle tartufaie naturali di tuber melanosporum vitt. dell’italia cebtrale. atti “il congresso internationale sul tartufo”, spoleto, 24-27 november 1988: 337-374, italy. bingham f.t. 1982. boron. (in:) a.l. page (ed.). methods of soil analysis. 2. agronomy mongraph. 2nd edition, madison. diamandis s. perlerou c. 2008. recent records of hypogeous fungi in greece. acta mycol. 43 (2): 139142. gee g.w., bauder j.w. 1986. particle-size analysis. (in:) a. klute (ed.) methods of soil analysis. soil science society of america; book series 5, madison: 383-411. gee w.g., or d. 2002. particle-size analysis. (in:) h.d jacob, t.g clarke, a.d warren (eds). methods of soil analysis. soil science society of america, madison: 151-184. 32 v. christopoulos et al. knudsen d., peterson, g.a., pratt p.f. 1982. lithium, sodium and potassium (in:) a.l. page (ed.). methods of soil analysis. 1. agronomy monograph. 2nd edition, madison. lulli l., primavera f. 2001. tuber magnatum pico: environment of growth. (in:) actes du ve congres international “science et culture de la truffe”, aix en provence, 4-6 mars 1999: 269-272, france. lindsay w.l., norvell w.a. 1978. development of a dtpa soil test for zinc, iron, manganese and copper. soil science society of america journal 42: 421-428. olsen s.r., cole c.v., watanabe f.s., dean l.a. 1954. estimation of available phosphorus in soils by extraction with sodium bicarbonate. us dept. of agric. circ. 939. piper c.s. 1942. soil and plant analysis. hassel press, adelaide, australia. poitou n. 1988. les sols truffiers francais. atti “il congresso internationale sul tartufo”, spoleto, 24-27 november 1988: 391-401, italy. raglione m., lorenzoni p., de simone c., monaco r., angius a. 1992. osservazioni sulle caratteristiche pedologiche di alcuni siti di tartufo nero pregiato (tuber melanosporum) in provincia di rieti. micologia e vegetazione mediterranea 7 (1): 211-224. raglione m., spadoni m., cavelli s., lorenzoni p., de simone c. 2001. les sols des truffieres naturelles de tuber melanosporum vitt. dans l’apennin central (italie). actes du ve congres international “science et culture de la truffe”, aix en provence, 4-6 mars 1999: 276-280, france. raglione m., owczarek m. 2005. the soils of natural environments for growth of truffles in italy. mycologia balcanica 3 (2): 209-216. walkley a., black i.a. 1934. an examination of the degtjareff method for determining soil organic matter, and a proposed modification of the chromic acid titration method. soil science 37: 29-38. http://dx.doi.org/10.1097%2f00010694-193401000-00003 2013-06-22t22:40:31+0100 polish botanical society sarcosoma globosum – an indicator of climate change? esteri ohenoja, maarit kaukonen and anna liisa ruotsalainen botanical museum, university of oulu, p.o.box 3000, fi-90014, oulu esteri.ohenoja@oulu.fi ohenoja e., kaukonen m., ruotsalainen a. l.: sarcosoma globosum – an indicator of climate change? acta mycol. 48 (1): 81–88, 2013. occurrence of a spring ascomycete, sarcosoma globosum has increased in finland during the last three decades. river banks and old spruce forests are its typical habitats. some of its habitats are damaged or even destroyed, however, because of forestry and building of roads and houses. it is a care-demanding fungus on the finnish red list, according to the iucn criteria, but its present status is being discussed. key words: ascomycota, sarcosomataceae, distribution, ecology, endangeredness introduction the genus sarcosoma belongs to the ascomycete order pezizales and to the family sarcosomataceae, s. globosum (schmidel) casp. being the only species in the genus in the temperate areas. it occurs in southern and central finland, usually in alluvial stands along rivers and brooks. it is rare and threatened in europe and one of the 33 fungal species proposed to the bern convention (dahlberg, croneborg 2003). s. globosum is also found in the eastern parts of n. america. description of the species sarcosoma globosum has big, sessile, roundish, barrel-like fruit bodies, 5-10 cm in diameter, dark brown-blackish, hymenium even, glossy, sterile part matt, velvety (fig. 1a). content of the fruit bodies is gelatinous. the genus name derives from greek and means ´fleshy body´. the fruit bodies turn flatter and more wrinkled when becoming older. dry weight percentage of the fungus is about three. asci are 8-spored, 250–300 × 35-38 μm in size, tips not amyloid with iodine. spores are hyaline, smooth, ellipsoid, 25-28 × 8-9 μm in size. paraphyses are filiform, acta mycologica vol. 48 (1): 81–88 2013 doi: 10.5586/am.2013.010 82 e. ohenoja et al. brown, brittle, as long as the asci and 2.5-4.0 μm wide (fig. 1b). as for the way of living of s. globosum, it is supposed to be saprophytic. ecology, phenology and distribution typical habitats of sarcosoma globosum are the forests along rivers and brooks (figs 1a, 2), often on alluvial shores, where the microclimate is favorable. the soil of flooded areas is rich in nutrients and moist also during dry springs. the fruit bodies often grow under aged spruces or pines among mosses and litter. s. globosum has been found in finland both on calcareous and acid ground, though it is considered to benefit from lime (martinsson, nitare 1986, tedebrand 1999). the plants and fungi reported in the vicinity of s. globosum are listed in table 1. s. globosum has been found in southern and central finland the northernmost locality lying on the latitude of 66 degrees (figs 3a-d, tab. 2). history of the occurrence of the species in finland has been presented by kosonen (1988). first finds of s. globosum are from 1915 and 1916, the locality rapolanharju (at sääksmäki, valkeakoski) being in southern finland (kivirikko 1916). this locality has not been continuously monitored, but it is protected, and the fungus was observed there e.g. in spring 1990. kuopio has been a good area, too (heikkilä, vauras 1982). the northernmost locality in finland, in kuusamo, has lately been visited only occasionally, but muhos, close to oulu, has been monitored regularly since the first discovery in 1973. fig. 1. sarcosoma globosum: a – on bank of the brook kalimeenoja in oulu. photo lassi kalleinen. 2012; b – microscopical characters (asci, spores and paraphyses) (muhos 2.vi.1973 m. ohenoja) microsc. esteri ohenoja 2013. a b sarcosoma globosum – an indicator of climate change? 83 fig. 2. typical habitat of sarcosoma globosum at muhos. photo esteri ohenoja 2012. table 1 plants and fungi found in the closest neighbourhood of sarcosoma globosum in finland. the species are in an order of the observed frequency in the stands in question vascular plants picea abies betula pubescens sorbus aucuparia pinus sylvestris juniperus communis rosa majalis prunus padus populus tremula alnus incana ribes spicatum lonicera xylosteum daphne mezereum salix caprea vaccinium vitis-idaea oxalis acetosella linnaea borealis orthilia secunda maianthemum bifolium luzula pilosa trientalis europaea vaccinium myrtillus convallaria majalis deschampsia flexuosa equisetum sylvaticum gymnocarpium dryopteris vaccinium myrtillus empetrum nigrum ssp. hermaphroditum silene dioica solidago virgaurea hieracium sp. paris quadrifolia rubus saxatilis melica nutans elymus caninus scirpus sylvaticus poa nemoralis diphasiastrum complanatum equisetum arvense matteuccia struthiopteris mosses hylocomium splendens pleurozium schreberi polytrichum commune dicranum polysetum rhytidiadelphus triquetrus brachythecium curtum rhodobryum roseum sphagnum sp. fungi microstoma protractum strobilurus esculentus auriscalpium vulgare 84 e. ohenoja et al. a b sarcosoma globosum – an indicator of climate change? 85 f ig s 3a -d . f in ds o f sa rc os om a gl ob os um in 1 91 520 12 ( ta bl e 2) . t he g ri d on t he m ap s is b as ed o n th e un if or m g ri d sy st em u se d (2 7° e ) in f in la nd . t he a re as o n th e m ap a re th e bi ol og ic al p ro vi nc es . c d 86 e. ohenoja et al. there are over 80 collects and observations of s. globosum from finland between 1915 and 2012. as for the phenology of the species (tab. 2), it has been found in southern finland in april-may, and in central finland in may-june, the main fruiting time being in the southern half of the country somewhat earlier than in the north. the species was found in southwestern finland once in december and in eastern finland in november. in 1985 when s. globosum was abundant in sweden (martinsson, nitare 1986) there was not a single find in finland, but the springs 1998 and 1999, when the fungus was very abundant in central sweden (tedebrand 1999) were good also in the southern part of finland, and it was observed first time also in simo, the north western most locality. the springs of 1990, 2001, 2005, 2008, and 2012 were good, especially in the northern part of the species´ distribution. the amount of fruit bodies has been even hundreds at some localities. there has been information about this curious fungus e.g., in some newspapers, and this has apparently increased knowledge of its occurrence in the country. table 2 the localities of sarcosoma globosum in finland abbreviations: 1b-4a forest vegetation zones (rassi et al. 2010) h herbarium of the university of helsinki, jyv herbarium of the jyväskylä university museum, kuo herbarium of kuopio natural history museum, oulu herbarium of the university of oulu, tur herbarium of the university of turku, tur-a herbarium of åbo akademi hemiboreal, oak zone (1b). lohja 22.4.1975 (h), 14.51987 (h); parainen 27.5.1978 (tur), 26.4.1989 (tur), 29.4.1989 (tur), 9.5.1989 (tur-a), 8.4.1990 (tur), 25.4.2000 (tur), 25.12.2000 (tur-a), 12.4.2001 (tur), 20.4.2001 (tur), 17.5.2001 (tur), .5.2006 (tur); perniö 11.5.1961 (tur); piikkiö 13.-14.5.1954 (tur), .3.1961 (h), 23.3.1961 (tur), 18.4.1961 (tur), 28.5.1961 (tur), 8.5.1963 (tur, oulu), 29.5.1967 (tur); rymättylä 14.4.1990 (tur); sauvo 9.4.1989 (tur); turku (kakskerta) 14.5.1980 (tur). southern boreal, sw finland and s ostrobothnia (2a). hattula 20.5.2000 (tur); hämeenkoski (koski) 28.5.1980 (oulu); pyhtää 22.4.2001 (h); säkylä 3.5.1998 (tur); valkeakoski (sääksmäki) 4.6.1915 (litt.), 7.5.1916 (litt.), 12.6.1980 (h), 1.6.1982 (?), 1.5.1987 (?), 27.5.1987 (tur), 21.5.1988 (tur), 29.4.1990 (oulu), 24.5.1998 (h, tur), 5.5.1998 (tur), 19.4.2005 (h). southern boreal, lake district (2b). heinävesi 12.5.1986 (h), 3.5.2001 (h); konnevesi 18.6.2008 (jyv); kuopio 16.4.1950 (kuo), 9.5.1954 (h), 23.5.1954 (kuo), 23.5.1973 (kuo), 26.5.1962 (tur), 22.5.1986 (kuo), 5.7.1994 (jyv), 5.6.1997 (tur), 17.5.1999 (kuo); maaninka 31.5.1998 (kuo); rautalampi 1.5.1990 (litt.), 9.6.1991 (kuo); savonlinna 6.6.2001 (h); 23.5.1992 (litt.); äänekoski 15.6.1982 (oulu). middle boreal, ostrobothnia (3a). liminka 9.5.2012 (obs.); muhos 2.6.1973 (oulu), 27.5.1988 (oulu), 24.5.1990 (oulu), 18.5.1993 (oulu), 28.5.1994 (oulu), 8.5.2010 (oulu), .6.2012 (obs.); oulu 21.5.2005 (oulu), 22.5.2012 (obs.), 23.5.2012 (obs.); oulu (haukipudas) 20.5.2005 (oulu); oulu (kiiminki) 27.5.2005 (oulu); revonlahti 8.5.2012 (obs.); raahe 10.5.2012 (obs.); tyrnävä 13.5.2012 (obs.). middle boreal, n karelia and kainuu (3b). hyrynsalmi 30.5.2001 (oulu), 12.5.2005 (oulu); kajaani 20.5.2008 (obs.), 25.5.2008 (obs.); nurmes 29.5.2005 (tur-a), 1.6.2005 (tur-a); puolanka 21.5.2005 (oulu). middle boreal, sw lapland (3c). simo .5.1999 (obs.), 15.5.2008 (oulu). northern boreal, kuusamo district (4a). kuusamo 19.6.1968 (h). russia karelia onegensis (kon). karhumäki 10.5.1943 (h). russia karelia australis (ka). viipuri 20.5.1891 (h). sarcosoma globosum – an indicator of climate change? 87 endangeredness sarcosoma globosum is, according to the criteria of iucn, a care-demanding fungus species in finland (rassi et al. 2010). it is included in ten red lists of european countries (dahlberg, croneberg 2003). in estonia it used to be very rare, but in 2005 and 2006 it appeared abundantly (kullmann 2011). it is very rare in norway, maybe because of the oceanic climate of the country, but in the semiarid, formerly pastured forests in sweden it has been abundant (tedebrand 1999). now it is, however, somewhat declining. in poland it is on the red list in the category of endangered species (wojewoda, ławrynowicz 2006). it is extinct from slovakia, germany and lithuania, but in latvia and norway it has been found again. it is obvious, that it is increasing in finland, but on the other hand it has lost habitats because of e.g. clearing of forest stands (fig. 4) and the human impact on the riversides. a protection plan has been prepared for one area of occurrence at muhos (kaukonen, ohenoja 1994), and that part of the river valley is now protected. also some other finnish localities are situated in nature reserves. discussion the distribution of sarcosoma globosum seems to be nemoral-boreal-montane. it always grows in same kind of habitats, characterized by the vicinity of rivers and brooks. though the fungus can sometimes be very difficult to find when hiding deep in the moss carpet, it is so striking in characters, that people have noticed it and also informed museums and researchers about it. why it seems to be more abundant now than before, the climate change with somewhat warmer winters and rainy springs could be one reason. martinsson and nitare wrote already in1986 that cold winters with less snow are not favorable for the fruiting of the fungus. for instance, there was a mild winter preceding a good yield of sarcosoma in finland in spring 1961. the latest winters have also been rather warm, with, however, much snow. based on the present knowledge, it would be appropriate to reevaluate the regional threat class of s. globosum on the finnish red list of species. acknowledgements. the authors thank the curators of the herbaria of h, iisalmi, jyv, kuo, tur, and tur-a, and many other friends, too, for sending data and collects. lassi kalleinen is warmly thanked for the photo and technical help. we congratulate our colleague and friend maria on her long standing, tough and productive mycological activity. references dahlberg a., croneborg h. 2003. 33 threatened fungi in europe. complementary and revised information on candidates for listing in appendix i of the bern convention. 82 pp. heikkilä h., vauras j. 1982. kuopion lehtokeskuksen suursienistä. savon luonto 14: 58-68. kaukonen m., ohenoja e. 1994. hytymaljakkaan (sarcosoma globosum) muhoksen esiintymien suojelusuunnitelma. oulun yliopiston kasvimuseo. 21 pp. kivirikko k.e. 1916. bulgaria globosa. meddelelser soc. fauna flora fennica 42: 144-145. 88 e. ohenoja et al. kosonen l. 1988. hytymaljakas (sarcosoma globosum), silmälläpidettävä kevätsieni. (summary: sarcosoma globosum in finland.) lutukka 4 (1): 3-6. kullmann b. 2011. limatünnik eestis. (in:) b. kullmann, t., liira, j., sammul, m. (toim.). haruldused eesti looduses. eesti looduseuurijate seltsi aastaraa mat 86: 9-17. martinsson k., nitare, j. 1986. bombmurklan, sarcosoma globosum, en hotad svamp. svensk bot. tidskr. 80: 169-184. stockholm. rassi p., hyvärinen e., juslén a., mannerkoski i. (eds). 2010. the 2010 red list of finnish species. ympäristöministeriö & suomen ympäristökeskus. helsinki, 685 pp. tedebrand j.-o. 1999. bombmurklan (sarcosoma globosum (j.c. schmidt: fr.) rehm i sverige. jordstjärnan 20 (2): 25-42. wojewoda w., ławrynowicz m. 2006. red list of the macrofungi in poland. (in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. w. szafer institute of botany, polish academy of sciences, kraków: 53-70. 2013-06-22t22:35:22+0100 polish botanical society contribution to the lichen biota of the pogórze wiśnickie foothills (carpathians) lucyna śliwa laboratory of lichenology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, l.sliwa@botany.pl śliwa l.: contribution to the lichen biota of the pogórze wiśnickie foothills (carpathians). acta mycol. 45(2): 219–230, 2010. the pogórze wiśnickie foothills are situated in close vicinity to the kraków agglomeration and is highly influenced by human activity. lichen studies in the area revealed 163 species so far. a current checklist of the lichen biota of the territory is provided with numerous new regional records, e.g., bacidina sulphurella, evernia prunastri, fuscidea pusilla, lecanora albellula, lepraria ecorticata, mycobilimbia epixanthoides, ramalina farinacea, r. fastigiata, ropalospora viridis, verrucaria praetermissa and v. tectorum. key words: lichens, biodiversity, lichenized fungi, new records, poland introduction the carpathians are among the best known areas in poland concerning the lichen biota (bielczyk 2006), with respective literature comprising ca 300 references (bielczyk 2003). the carpathian foothills, however, though being close to the centres of lichenological investigations of southern poland, remain poorly explored both historically and geographically. among the foothills the most thoroughly explored areas are: pogórze rożnowskie and ciężkowicke (kozik 1970, 1976, 1977; czwórnóg, śliwa 1995), pogórze spisko-gubałowskie (kiszka 1985), pogórze przemyskie (kiszka, piórecki 1991; kiszka 2002a, b). fragmentary data originate from pogórze bukowskie (rydzak 1955), pogórze wielickie (kiszka 1996a), pogórze dynowskie (krzewicka, śliwa 2000), and pogórze śląskie (leśniański 2001) foothills. the latter regions as well as others (e.g., pogórze strzyżowskie, rzeszowskie and jasielskie foothills) are in urgent need of complex investigations especially with rapid changes of environmental conditions caused by human activity and the influence of changing climate. due to these same reasons some of the early investigations require reexamination. acta mycologica vol. 45 (2): 219–230 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 220 l. śliwa in the years 1998-2000 a lichen survey was carried out aimed at a more comprehensive exploration of the pogórze wiśnickie foothills with special emphasis on protected areas. the project included lichenological training for students of the jagiellonian university and resulted in some joint publications (śliwa et al. 2001; śliwa, krzewicka 2004). part of the results were also published by stolarczyk (2003). this paper presents a part of a summary of the research as a contribution to the knowledge of the lichen biota of this interesting area highly influenced by human activity. study area the carpathian foothills form a transitional area that is located between the beskid in the south and the sub-carpathian basins in the north, and the landscape is affected by three main subsystems: abiotic, biotic and anthropogenic (drużkowski 1998). the pogórze wiśnickie foothills constitute the easternmost edge of the pogórze zachodniobeskidzkie foothills of the western carpathians (kondracki 1989, 2001). neighbouring areas are the pogórze wielickie foothills to the west and the pogórze rożnowskie foothills to the east. the territory includes several protected areas (e.g., the bukowiec nature reserve, kamień grzyb and kamienie brodzińskiego pro-kamień grzyb and kamienie brodzińskiego pro-protected sandstone tors) as well as areas of special concern (wiśnicko-lipnicki land-as well as areas of special concern (wiśnicko-lipnicki landscape park). in general it is rich in natural values represented by forested hills and unique landscape formations such as sandstone tors. forest communities occupying the area were characterised by stachurska (1998a, b). on the other hand the pogórze wiśnickie foothills is under the visible impact of human activity; some parts of the area are strongly urbanized and influenced by industrial and transportation emissions of krakow and surrounding towns. details of the environmental transformation of the whole area of the carpathians foothills due to natural and anthropogenic factors is presented by drużkowski (1998), who evaluated the transformation as moderate at present. material and methods lichens were collected in the years 1998-2000 at 32 sites located in the fe atpol gird square system (acc. to cieśliński, fałtynowicz 1993). each 100×100 km (fe) plot was divided into 10×10 km units numbered from 00 to 99, and then into 2×2 km subunits also numbered from 00 to 99 − the numbers follow one the other in the list of collecting sites. all habitats and substrates were explored. lichens were identified using routine microscopic and laboratory techniques. when necessary the tlc analyses was performed in solvent system a or/and c (methods followed orange et al. 2001). voucher specimens are available at kra and/or kram herbaria. nomenclature basically follows santesson et al. (2004) and diederich et al. (2010). contribution to the lichen biota 221 lichen collecting sites in the pogórze wiśnickie foothills: 1 − cichawka village, s slope of a hill along the road, fe 8233; forest with pinus sylvestris and quercus robur, and road side trees, 8 may 1998, l. śliwa collection numbers 539-561; 2 − cichawka village, near the church, fe 8233; stream valley with alnus glutinosa, rocks and stones, 8 may 1998, l. śliwa 562-592; 3 − wieruszycka village, along stradomka stream, fe 8232, shrubs with alnus glutinosa, 19 may 1998, l. śliwa 593-614; 4 − cichawka stream headwaters, fe 8234, mixed forest with fagus sylvatica, carpinus betulus, pinus sylvestris and abies alba, 19 may 1998, l. śliwa 615-662; 5 − królówkaskotnica village, along stream, fe 8331, mixed forest and concrete constructions; 2 june 1998, l. śliwa 663-728; 6 − królówka-uzbornia village, fe 8321, trees along road side and stream, 2 june 1998, l. śliwa 663-728; 7 − e part of łapanów village, road to muchówka, fe 8231, road side trees of fraxinus excelsior, 3 june 1998, l. śliwa 662-796; 8 − łapanów-rogatka village, fe 8232, trees along stream and road side trees of populus spp., 3 june 1998, l. śliwa 797-837; 9 − wola wieruszycka village, fe 8222, salix spp. trees along stream and fruit trees, 17 june 1998, l. śliwa 838-873; 10 − chrostowa góra mt., fe 8212, mixed forest with quercus robur, fagus silvatica and pinus sylvestris, 17 june 1998, l. śliwa 874-880; 11 − chrostowa village, fe 8212, road side trees, 17 june 1998, l. śliwa 881-898; 12 − dąbrowica village, park by the school, fe 8211, 17 june 1998, l. śliwa 899-910; 13 − forest between dołuszyce and pogwizdów and kobylany villages, s slope, fe 7341, mixed forest with fagus sylvatica, quercus robur, carpinus betulus, betula pendula, and pinus sylvestris, 21 april 1999, l. śliwa 975-992; 14 − road from dołuszyce to pogwizdów village, along a stream, fe 7340, young forest with alnus glutinosa, fraxinus excelsior and quercus robur, 21 april 1999, l. śliwa 993-1016; 15 − bukowiec nature reserve, alt. 430-460 m, fe 8442, mixed forest and shaded streams, 21 may 1999 and 22 sept. 1999, l. śliwa 1017-1075; 16 − tymowa górna village, fe 9342, road side trees, 22 sept. 1999, l. śliwa 1076-1082; 17 − lipnica murowana village, road to nowy wiśnicz town, fe 8334, salix spp., 1 march 2000, l. śliwa 1083; 18 − stary wiśnicz town, road junction, fe 8303, concrete post, 24 march 2000, l. śliwa 1084; 19 − lipnica górna village, road to rajbrot, fe 8344, betula pendula, 24 march 2000, l. śliwa 1085; 20 − e part of kobyle village, road side trees, fe 8400, populus spp., 24 march 2000, l. śliwa 1086-1087; 21 − krasa góra village, road side, fe 8420, 24 march 2000, l. śliwa 1088; 22 − muchówka village, road to żegocina, fe 8342, 24 march 2000, l. śliwa 1089-1093; 23 − nw slope of the hill “409 m a.s.l” near mt. paprotna góra, fe 8342, mixed forest with rocks, 7 may 2000, l. śliwa 1094-1104; 24 − hill by road to rajbrot village, near mt. paprotna górna, fe 8342; mixed forest with pinus sylvestris and betula pendula, 7 maj 2000, l. śliwa 1105-1114; 25 − “kamień grzyb” stone in bigorzówka village near raciechowice, fe 9102, sandstone, 7 may 2000, l. śliwa 1115-1126; 26 − “diabelski kamień” stone in smykan village near szczyrzyce, fe 9122, sandstone, 7 may 2000, l. śliwa 1127-1159; 27 − forest s of sobolów village, fe 8215, mixed forest with quercus robur, fagus sylvatica and picea abies, 5 june 2000, l. śliwa 1160-1182; 28 − mt. góra łysa between zonia and nieprzaśna villages, near quarry, fe 8214, forest with fagus sylvatica and pinus sylvestris, 5 june 2000, l. śliwa 1183-1190; 29 − wichraź village near zagrody, stream valley, fe 8224, forest ridge, 20 june 2000, l. śliwa 1191-1212; 30 − sieradzka village, stream valley, fe 8223, forest with fraxinus excelsior, alnus glutinosa and carpinus betulus, 20 june 2000, l. śliwa 1213-1234; 31 − dziekanowice village, along raba river, fe 8014, 222 l. śliwa sandstone outcrops, 29 june 2000, l. śliwa 1235-1257; 32 − leszczyna village, w of junction with the road to rozstajnie village, fe 8234, road side trees, mainly salix spp., 30 june 2000, l. śliwa 1258-1267. results and discussion the checklist of taxa presented below (tab. 1) summarizes all known data concerning the lichen biota of the pogórze wiśnickie foothills. table 1 lichenized fungi recorded in the pogórze wiśnickie foothills until the present source of information: 1 – śliwa et al. (2001); 2 – stolarczyk (2003); 3 – śliwa & krzewicka (2004); 4 – herbarium material, leg. śliwa (coll. no.). bark of trees: ab – abies alba, ap – acer platanoides, ae – aesculus hippocastanum, al – alnus glutinosa, b – betula pendula, cb – carpinus betulus, fs – fagus sylvatica, fr – fraxinus excelsior, jr – juglans regia, ma – malus domestica, lar – larix decidua, ps – pinus sylvestris, po – populus spp., pr – prunus spp., qr – quercus robur, sa – salix spp., tc – tilia cordata. name of species substrate source of information acarospora fuscata (schrad.) th.fr. sandstone rocks, concrete 1; 2 acrocordia gemmata (ach.) a. massal. po 2; 4 (609, 1260) amandinea punctata (hoffm.) coppins & scheid. po, tc, fr, sa, qr, b, ma, jr 2; 4 (540b, 547, 605, 597, 720, 735, 751, 772, 826a, 832a, 838, 847, 883, 884, 900) anisomeridium polypori (ellis & everh.) m.e. barr po 2; 4 (857, 996, 1216b, 1086) arthonia radiata (pers.) ach. fs, al, cb 4 (564, 659, 703, 705, 998, 1005b, 1216a, 1221b, 1223, 1228) arthonia spadicea leight. cb 4 (563) aspicilia calcarea (l.) mudd calcareous stones, concrete, tile 2 bacidia trachona (ach.) lettau sandstone rocks 1 bacidina sulphurella (samp.) comb. ined. fs, cb 4 (598, 612, 1060, 1061) baeomyces rufus (huds.) rebent. soil, sandstone rocks 1; 2; 3; 4 (621, 977, 1112) bilimbia sabuletorum (schreb.) arnold sandstone, concrete, bryophytes 2 buellia griseovirens (sm.) almb. po, ma 2; 4 (726, 754, 849b, 1003) caloplaca citrina (hoffm.) th.fr. concrete, sandstone rocks, asbestos tile 2; 4 (671, 795, 1093) caloplaca vitellinula auct. concrete 2 (needs rev.) caloplaca decipiens (arnold) blomb. & forssell concrete, asbestos tile, tile 2; 4 (683, 714, 1192) caloplaca holocarpa (ach.) a.e. wade concrete, sandstone rocks, tile 2; 4 (712) caloplaca velana (a. massal.) du rietz concrete 4 (674, 863b, 866) caloplaca saxicola (hoffm.) nordin concrete 2 candelaria concolor (dicks.) stein po 2; 4 (665a) candelariella aurella (hoffm.) zahlbr. concrete, sandstone rocks, tile 2 candelariella coralliza (nyl.) h. magn. sandstone rocks 2 candelariella reflexa (nyl.) lettau ma, po, sa, fr, tc, ae 2; 4 (599, 665a, 740, 818, 1007, 1077) candelariella vitellina (hoffm.) müll. arg. sandstone rocks, tile 2; 4 (713, 724) candelariella xanthostigma (ach.) lettau po, sa, tc, fr, ma, jr 2; 4 (1078) contribution to the lichen biota 223 chaenotheca chrysocephala (ach.) th.fr. sa 4 (684, 848) chaenotheca ferruginea (turner & borrer) mig. al, fs, ps 2; 3; 4 (677, 1008, 1010) chaenotheca furfuracea (l.) tibell soil, roots of trees 4 (575, 1198) chaenotheca xyloxena nádv. wood 3 cladonia caespiticia (pers.) flörke soil 1; 2; 4 (551, 556, 1161, 1172) cladonia chlorophaea (sommerf.) spreng. soil, sandstones, ps, b, sa, fr, ma 1; 2; 4 (988) cladonia coniocraea (flörke) spreng. soil, wood, bark of trees 1; 2; 3; 4 (585, 641, 855, 992) cladonia digitata (l.) hoffm. ps 1; 2 cladonia fimbriata (l.) fr. soil 2; 4 (733, 831, 985, 1181, 1231) cladonia macilenta hoffm. b, ps; wood, soil 1; 2; 3; 4 (890) cladonia ochrochlora flörke ps; wood 1; 2; 3 cladonia pleurota (flörke) schaer. sandstone rocks 1 cladonia parasitica (hoffm.) hoffm. soil 2; 3 (needs rev.) cladonia pyxidata (l.) hoffm. soil 2 cladonia rei schaer. soil 2 (needs rev.) cladonia squamosa hoffm. ps 2 cladonia subulata (l.) f.h. wigg. soil 2 coenogonium pineti (ach.) lücking & lumbsch b, qr 2; 3 (sub dimerella diluta); 4 (557b, 994b, 1109, 1191) collema tenax (sw.) ach. soil 4 (1089) dibaeis baeomyces (l.f.) rambold & hertel soil 2; 4 (560, 572) diploschistes scruposus (schreb.) norman sandstone rocks 1 evernia prunastri (l.) ach. po 4 (545, 781, 1267) fuscidea pusilla tønsberg al 4 (704, 1220a, 1221d) graphis scripta (l.) ach. fs, cb 1; 3; 4 (567, 649, 650, 661b, 696, 697, 702, 1005a, 1225a, 1230b) hypocenomyce caradocensis (nyl.) p. james & gotth. schneid. ps 2; 3 hypocenomyce scalaris (ach.) m. choisy b, fs, fr, ma, ps; wood 2; 3; 4 (552, 648, 727, 776, 825, 854, 876, 909b, 975, 1013, 1098, 1168) hypogymnia physodes (l.) nyl. ps, po, fr, qr, ma, sa, pr; tile 2; 3; 4 (576, 602, 634, 687, 743, 817, 851c, 1227) hypogymnia tubulosa (schaer.) hav. ma 2; 4 (1217) imshaugia aleurites (ach.) s.l.f. meyer b 4 (1104) lecania cyrtella (ach.) th.fr. al, sa, b 4 (755, 816, 821a) lecanora albellula nyl. po, sa, fs, qr, ma 4 (601, 610, 678, 746, 757, 1173a) lecanora albescens (hoffm.) flörke concrete 2; 4 (1202) lecanora carpinea (l.) vain. fr, sa 4 (681, 815) lecanora chlarotera nyl. po 4 (736) lecanora conizaeoides crombie ps, b, sa, qr, po, fs, al, ap, lar, cb, ma, pr; wood 1; 2; 3; 4 (553, 565, 614, 625, 652, 676, 688a, 773, 819, 862, 888, 904, 909a, 986) lecanora dispersa (pers.) sommerf. concrete, sandstone rocks, bark of trees, asbestos tile, tile 2 (incl. l. semipallida); 4 (760, 796, 867, 1091, 1201) lecanora expallens ach. po, sa, ap, fr, tc, qr, ma; asbestos tile 2 (needs rev.) lecanora hagenii (ach.) ach. wood, concrete 4 (794, 1164a, 1200) lecanora muralis (schreb.) rabenh. sandstone rocks 2; 4 (692) lecanora polytropa (hoffm.) rabenh. sandstone rocks 1 lecanora pulicaris (pers.) ach. sa, al, qr, ma; wood 2; 3; 4 (655, 680) lecanora saligna (schrad.) zahlbr. po, sa, fs, qr, ma; wood 2; 4 (624, 672, 834, 1014, 1195, 1257, 1264) table 1 — cont. 224 l. śliwa lecanora semipallida h. magn. concrete 4 (701, 711, 729a, 1084, 1090, 1164) lecanora symmicta (ach.) ach. wood 4 (851a) lecanora varia (hoffm.) ach. wood 2; 4 (695) lecidea fuscoatra (l.) ach. sandstone rocks 1 lecidella elaeochroma (ach.) m. choisy sa, b 2 lecidella stigmatea (ach.) hertel & leuckert concrete, tile 2; 4 (653, 710, 797, 832b, 833a, 845, 1092) lepraria borealis lothander & tønsberg sandtone rocks, bryophytes 4 (1119a) lepraria caesioalba (de lesd.) j.r. laundon sandtone rocks, soil, bryophytes 1; 4 (1119b) lepraria eburnea j.r. laundon sa 4 (774a, 1222, 1262) lepraria ecorticata (j.r. laundon) kukwa sandstone rock 4 (1132) lepraria elobata tønsberg al, sa, ma, b, fs; soil, rocks 3; 4 (557a, 586, 643, 669, 774b, 813, 982, 1096, 1110, 1113, 1162, 1188, 1204, 1252) lepraria incana (l.) ach. po, fr, sa, fs, b, tc; sandstone rocks 1; 3; 4 (676, 689, 899, 987, 1082, 1133, 1144, 1207, 1263) lepraria jackii tønsberg bryophytes 4 (1169, 1171) lepraria lobificans nyl. sa, tc, al, fs, ps; wood, sandstone rocks 1; 3; 4 (581, 608, 734, 774c, 839, 978, 995, 1094, 1105, 1114, 1205, 1225b, 1230a, 1256) lepraria membranacea (dicks.) vain. sandstone rocks 1; 2; 4 (1136, 1147b, 1238) lepraria neglecta (nyl.) lettau sandstone rocks 1; 4 (1119c, 1147a) lepraria vouauxii (hue) r.c. harris sa, po, ma 4 (546, 632, 764, 836, 887, 893) melanelixia fuliginosa (duby) o. blanco et al. po, ap, ae, ma 2; 4 (647, 678, 715) melanelixia subargentifera (nyl.) o. blanco et al. sa 4 (778, 1258) melanelixia subaurifera (nyl.) o. blanco et al. qr, ae 2; 4 (567) melanohalea elegantula (zahlbr.) o. blanco et al. sa 2 (sub melanelia incolorata) melanohalea exasperatula (nyl.) o. blanco et al. tc, sa, fr, ma; tile 2; 4 (593, 890) micarea botryoides (nyl.) coppins bryophytes, wood 3 (cf. czarnota 2007); 4 (592, 699) micarea denigrata (fr.) hedl. sandstone, wood 2; 3; 4 (729b, 1097) micarea lignaria (ach.) hedl. b; soil 4 (577, 1102) micarea micrococca (körb) coppins cb, ps 4 (639c, 1232) micarea misella (nyl.) hedl. rotting wood 3; 4 (1182, 1197b) micarea peliocarpa (anzi) coppins & r. sant. sandstone rock 1 micarea prasina fr. rotting wood 3; 4 (615b, 627b, 679, 1108a) micarea viridileprosa coppins & v.d. boom wood 3 (cf. czarnota 2007); 4 (1108b) mycobilimbia epixanthoides (nyl.) hafellner & türk sa 4 (623, 666, 840, 850, 1081, 1083, 1173b) mycoblastus fucatus (stirtan) zahlbr. b, qr 2; 4 (656) parmelia omphalodes (l.) ach. sandstone rocks 1 parmelia saxatilis (l.) ach. sandstone rocks, ma 1; 2; 3; 4 (584, 1009, 1012, 1103) parmelia sulcata taylor po, sa, fr, al, tc, qr, ma; tile 2; 4 (571, 594, 628, 717, 752, 770, 837a, 849a, 859, 1004) parmelina tiliacea (hoffm.) hale tc, po 2; 4 (768, 802) parmeliopsis ambigua (wulfen) nyl. ma, al 2 pertusaria albescens (huds.) m. choisy & werner ap, sa 2 table 1 — cont. contribution to the lichen biota 225 phaeophyscia nigricans (flörke) moberg po; concrete, tile 2; 4 (741) phaeophyscia orbicularis (neck.) moberg po, sa, tc, ae, fr, cb, jr, ma; concrete, asbestos tile 2; 4 (603, 747, 767, 780a, 873, 892, 898, 905, 1183) phaeophyscia sciastra (ach.) moberg concrete 4 (693) phlyctis argena (spreng.) flot. tc, fr, ma 1; 2; 4 (542, 635, 790, 812, 851b, 901) physcia adscendens (fr.) h. olivier po, fr, sa, ma, cb, jr, ae, tc, al; concrete 2; 4 (539, 640, 706, 730, 748, 749, 771, 807, 895, 1015, 1079) physcia caesia (hoffm.) fürnr. concrete, tile 2; 4 (758, 786) physcia dubia (hoffm.) lettau concrete 2; 4 (737) physcia stellaris (l.) nyl. po 2; 4 (792) physcia tenella (scop.) dc. po, tc, fr, sa, ma, jr 2; 4 (595, 596, 620, 691, 719, 777, 1226) physconia detersa (nyl.) poelt po 4 (779a) physconia distorta (with.) j.r. laundon po 4 (763, 783) physconia enteroxantha (nyl.) poelt po, sa 2; 4 (742, 779b) physconia grisea (lam.) poelt po, sa, fr 2; 4 (780b) physconia perisidiosa (erichsen) moberg po, sa 2; 4 (1011) placynthiella dasaea (stirt.) tønsberg wood, ps; sandstone rocks 2; 3; 4 (1170, 1178) placynthiella icmalea (ach.) coppins & p. james sa, al, b; wood 1; 2; 3; 4 (1006, 1166) placynthiella uliginosa (schrad.) coppins & p. james ma, po, fr; wood 2; 3; 4 (980) placynthium nigrum (huds.) gray sandstone rocks 2; 4 (664) platismatia glauca w.l. culb. & c.f. culb. wood 4 (894) porina aenea (wallr.) zahlbr. fs 2; 3; 4 (639a, 661a, 993, 1221c, 1224) porpidia tuberculosa (sm.) hertel & knoph sandstone rocks 2 protoblastenia rupestris (scop.) j. steiner concrete, asbestos tile 2 protoparmelia hypotremella herk, spier & v.wirth b, fr 2; 4 (789) pseudevernia furfuracea (l.) zopf ma; sandstone rocks, tile 1; 2; 4 (573b, 756, 897, 1218) punctelia subrudecta (nyl.) krog sa 2; 4 (630, 787, 902) pyrenula nitida (weigel) ach. fs 4 (1177a) ramalina farinacea (l.) ach. fr 4 (791) ramalina fastigiata (pers.) ach. po 4 (803) ramalina pollinaria (westr.) ach. sandstone rocks 2 ropalospora viridis (tønsberg) tønsberg cb 4 (619, 1221e) sarcogyne regularis körb. concrete 2; 4 (694, 1206) scoliciosporum chlorococcum (stenh.) vězda po, sa, ma, fs, lar, fr, ap, b, ps, qr, ab, cb, pr 1; 2; 3; 4 (561, 638, 725, 739, 759, 766, 853, 877a, 981) scoliciosporum umbrinum (ach.) arnold sandstone 2 trapelia coarctata (sm.) m. choisy sandstone rocks, pebbles 2; 3; 4 (644, 654, 660, 744, 991, 1180, 1184) trapelia involuta (taylor) hertel sandstone rocks 1; 3 trapelia obtegens (th.fr.) hertel sandstone rocks 1; 2 trapelia placodioides coppins & p. james sandstone rocks, tile 1; 2 trapeliopsis flexuosa (fr.) coppins & p. james qr, ma, b; wood 2; 3; 4 (1163) trapeliopsis gelatinosa (flörke) coppins & p. james soil 1 trapeliopsis granulosa (hoffm.) lumbsch soil, wood 3 trapeliopsis pseudogranulosa coppins & p. james wood, soil, bryophytes 2; 3; 4 (1001) trapeliopsis viridescens (schrad.) coppins & p. james rotting wood 2; 3 (needs rev.) table 1 — cont. 226 l. śliwa umbilicaria deusta (l.) baumg. sandstone rocks 1; 2 umbilicaria hirsuta (westr.) hoffm. sandstone rocks 1; 2 verrucaria caerulea dc. sandstone rocks 2 (sub v. glaucina, needs rev.) verrucaria muralis ach. concrete, pebbles 4 (732, 784) verrucaria nigrescens pers. tile, concrete, pebbles 2 (needs rev.); 4 (728) verrucaria praetermissa (trevis.) anzi stone in stream beds 4 (1043) verrucaria tectorum (a. massal.) körb. sandstone rock 4 (868) verrucaria velana (a. massal.) zahlbr. concrete, stones 2 xanthoparmelia conspersa (ach.) hale sandstone rocks 1; 2 xanthoparmelia loxodes (nyl.) o. blanco et al. sandstone rocks 1; 2 xanthoparmelia stenophylla (ach.) ahti & d. hawksw. sandstone rocks 1; 2 (sub x. somolënsis, needs rev.) xanthoria candelaria (l.) th.fr. po, fr 2; 4 (788) xanthoria elegans (link.) th.fr. concrete 2; 4 (1255) xanthoria fallax (hepp) arnold po; asbestos tile 2; (793) xanthoria parietina (l.) th.fr. po, ma, fr, tc; concrete 2; 4 (611, 738, 769, 810, 871, 889) xanthoria polycarpa (hoffm.) rieber po, sa, fr 2 the list of taxa presented summarizes all reported data (163 species so far) but most of all provides many new regional records. the newly recorded species represent various groups of lichens both in terms of taxonomy and ecology: members of large crustose genera that have been recently revised using modern approaches – bacidia s.l., caloplaca, lecanora, lepraria, micarea, verrucaria; widespread sterile species reported as frequent or common elsewhere, such fuscidea pusilla, mycobilimbia epixanthoides, ropalospora viridis; and endengered species included in the red list of extinct and threatened lichens in poland (cieśliński et al. 2003) – evernia prunastri, ramalina farinacea, r. fastigiata. several of the species deserve special attention. bacidina sulphurella was reported only from a few sites in poland until now: góry sowie mts, on acer pseudoplatanus and puszcza knyszyńska forest, vicinity of czarna białostocka, on carpinus betulus (brand et al. 2009), and warszawa city, “las bielański” nature reserve, on wood (kubiak et al. 2010). however, it is considered a widespread species in the country (kubiak et al. 2010). the species represents the b. arnoldiana group, which is distingushed by a finely granular thallus entirely covered by goniocysts, rather large and flat apothecia, with a greyish-brown disc with a slight violet hue, and paler orangebrown and raised margins, and a typically dark brown to red-brown hypothecium reacting k+ dark brown. revision of the material referred to as b. arnoldiana in western europe and macaronesia by brand et al. (2009) demonstrated that two species can be recognized on the basis of the size of pycnidia and more particularly on the shape of the conidia: b. arnoldiana (körb.) v. wirth & vězda characterised by filiform, arched or curved (rarely almost straight) conidia, and b. sulphurella characterised by filiform conidia, curved or not, but always with at least one end strongly hooked and slightly enlargered. moreover, the authors discovered that in addition to morphology the species differ in ecology; the former one is saxicolous and the latter corticolous. lecanora albellula [syn. l. piniperda körb.] was being reported during the last decade with increasing frequency as a result of a broader concept of the species table 1 — cont. contribution to the lichen biota 227 presented by printzen (2001). the species belongs to the l. saligna-related taxa that are characterised by the presence of usnic and/or isousnic acids and corticate amphithecium (traditionally named the l. varia group). additionally it is distinguished by apothecia rounded to flexuose, usually densely crowded, more rarely single or in small groups, sessile, 0.4−0.6(−0.9) mm diam.; disc light ochre to reddish-brown, matt, finely whitish pruinose, flat to modertely convex; margin weakly prominent when young, persistent or level with disc or often excluded in old apothecia. the most diagnostic character of the species is the apothecial granules that are abundant in the epithecium and densely obscure the whole area of amphithecial cortex. the granules are bright in polarized light and dissolve rapidly in koh. lecanora albellula can be mistaken for l. subintricata (nyl.) th. fr. and specimens with dark apothecial discs can also be confused with l. saligna (printzen 2001). moreover, the study of the polish collections of the l. varia group indicated the taxon l. saligna var. sarcopis (ach.) hillm. at least pro parte in fact represents l. albellula. the status of the taxa are in urgent need of further investigations. lepraria ecorticata has been recently reported for the first time from continental europe by kukwa (2006). the author presented records of the species from poland and the czech republic. in poland lepraria ecorticata was noted from równina bielska plain, kaszuby landlake and warmia in the north, and from pogórze karko-karkonoskie foothills, góry sowie mts, beskid wyspowy mts and gorce mts in the south. the species is characterized by a thick, not stratified thallus with most modullary hyphae and soredia well separated from one another. l. ecorticata is very similar in appearance to l. elobata but produces usnic acid in addition to zeorin. it resembles also sorediate species of lecanora that contain usnic acid as well, e.g., l. expallens, l. compallens herk & aptroot and l. leuckertiana (cf. kukwa 2006). verrucaria praetermissa is an amphibious species occurring exclusively in humid habitats and characterized by a pale green thallus with pinkish white prothallus. the species occupies large siliceous stones emerging above the water surface in streams but occasionally inundated. it is relatively frequently recorded both in lowland and mountainous areas of the country (e.g., kiszka 1996b; czarnota 2000; zalewska 2000; czyżewska et al. 2001, 2002; bielczyk 2003; cieśliński 2003; krzewicka 2006, 2009). verrucaria tectorum is a member of the v. nigrescens complex. it is a saxicolous species occupying calcareous rocks and human-made calcium-containing substrata. the species was rarely reported from poland (rehman 1879; kiszka, kościelniak 1996; sparrius 2003) but a recent revision of the genus in poland demonstrated this poorly known species was often mistakenly reported as v. nigrescens pers. (krzewicka, unpubl.). the quoted treatment yielded numerous new records of the species from the area of the whole country. 228 l. śliwa conclusion preliminary inventory of the lichens of the pogórze wiśnickie foothills indicates considerable diversity in spite of the high infl uence of human activity on the land-high influence of human activity on the landscape and nature of the carpathians foothills. additionally such a large representa-. additionally such a large representation of newly reported species belonging to various lichen groups, both in terms of taxonomy and ecology, indicate that further herbarium and field investigations will yield more interesting discoveries. finally species that urgently require special attention are the lichenicolous fungi. so far only three of the most common species have been reported from the area: athelia arachnoidea (stolarczyk 2003), lichenoconium erodens and l. lecanorae (stolarczyk 2003; śliwa, krzewicka 2004). acknowledgements. it is a great pleasure and honour to dedicate this paper to professor barbara gumińska with sincere thanks for her kindness and warm support during my student and academic career at the institute of botany of the jagiellonian university in the years 1986-2003. i am grateful to beata krzewicka for friendly and most valuable support during the course of the study. martin kukwa (gdańsk), paweł czarnota (niedźwiedź), beata krzewicka and karina wilk (kraków) are thanked for the determination or revising of some critical taxa, dariusz kubiak for sharing his ideas and personal comments on taxonomy of bacidia arnoldiana group, and the following students for accompany me on some field trips: aneta sosin, karina palka (now karina wilk), barbara haduch, michał węgrzyn, jerzy smykla and piotr stolarczyk (kraków). sincere thanks are due to krystyna czyżewska (łódź) for valuable comments on a paper proposal and to the anonymous reviewer for suggestions on the manuscript. the study was partly supported by the ministry of science and higher education, grant no. nn 304 170539. references bielczyk u. 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(heinrich z., wojewoda w.) columnocystis abietina (pers. ex fr.) pouzar (corticiaceae) in polish carpathians. fragm. flor. geobot. 20 (3): 397-403 (in polish with english summary). – macromycetes of the ojców national park. i. the flora. acta mycol. 10 (2): 181-265 (in polish with english summary). – basidiodendron caesiocinereum (höhn. et litsch.) luck-allen (tremellales) in poland. fragm. flor. geobot. 20 (3): 405-410. – exidiopsis grisea (pers.) bourd. et maire sensu reid (1970) in poland. fragm. flor. geobot. 20 (4): 547-551. – marinus anton donk (1908–1972). wiadomości botaniczne 18 (3): 165-167. – phot.: tabl. ii. 1 czarcia miotła na carpinus betulus l. spowodowana przez taphrina carpini (rostr.) johans. prądnik korzkiewski, 22 vi 1972. (in:) b. sałata, workowce (ascomycetes). szpetkowe (taphrinales). (in:) j. kochman, a. skirgiełło (eds). flora polska. rośliny zarodnikowe polski i krajów ościennych. 6. grzyby. państwowe wydawnictwo naukowe, warszawa–kraków. – phot.: tabl. ii. 2 – gałąź carpinus betulus l. zniekształcona przez taphrina carpini (rostr.) johans. prądnik korzkiewski, 22 vi 1972. (in:) b. sałata, workowce (ascomycetes). szpetkowe (taphrinales). (in:) j. kochman, a. skirgiełło (eds). flora polska. rośliny zarodnikowe polski i krajów ościennych. 6. grzyby. państwowe wydawnictwo naukowe, warszawa–kraków. 1975. macromycetes of the ojcównational park. ii. phytosociological, ecological and geographical characterization. acta mycol. 11(2): 263-309 (in polish with english summary). – albert pilát (1903-1974). wiadomości botaniczne 19 (1): 3-5. – gatunki rodzaju tremella pasożytujące na grzybach. wiadomości botaniczne 19 (2): 119-123. – bogumił eichler (1843-1905) jako mikolog. wiadomości botaniczne 19 (3): 157-161. 1976. (heinrich z., wojewoda w. ). the effect of fertilization on a pine forest ecosystem in an industrial region. ekologia polska 24 (3): 319-330. – (kućmierz j., wojewoda w.). sto lat badań mikologicznych w dolinie ojcowskiej. wiadomości botaniczne 20 (2): 97-104. – (turowska i., podbielkowski z., wojewoda w.). rośliny zarodnikowe (2nd ed). akademia medycz-medyczna, kraków. – polish dacrymycetales. 1. dacrymyces estonicus raitv. and d. ovisporus bref. fragm. flor. geobot. 22 (3): 395-400. – disappearance of sites with macromycetes in poland. phytocoenosis 5 (3/4): 377-386 (in polish with english summary). – feliks berdau (1826-1895) jako mikolog. wiadomości botaniczne 20 (3): 143-145. – michael e., hennig b. & kreisel h. 1975. handbuch für pilzfreunde. 6. wiadomości botaniczne 20 (2): 136-137 (a review; in polish). – prochacki h. 1975. podstawy mikologii lekarskiej. wiadomości botaniczne 20 (2): 137-139 (a review; in polish). – singer r. 1975. agaricales in modern taxonomy. wiadomości botaniczne 20 (3): 192-193 (a review; in polish). – farr m. l. 1973. an annotated list of spegazzini’s fungus taxa. wiadomości botaniczne 20 (3): 193-194 (a review; in polish). – karsten p. a. 1973. collected mycological papers. 1-4. wiadomości botaniczne 20 (4): 272-274 (a review; in polish). 1977. podstawczaki (basidiomycetes), trzęsakowe (tremellales), uszakowe (auriculariales), czerwcogrzybowe (septobasidiales). (in:) j. kochman, a. skirgiełło (eds). flora polska. rośliny zarodnikowe polski i ziem ościennych. 8. grzyby. państwowe wydawnictwo naukowe, warszawa– kraków. – polish dacrymycetales. 2. calocera furcata (fr.) fr. fragm. flor. geobot. 23 (1): 113-117. – grzyby wielkoowocnikowe. in: k. zabierowski (ed.). przyroda ojcowskiego parku narodowego. studia naturae ser. b. 28: 161-181. – stanisław chełchowski (1866-1907) jako mikolog. w siedemdziesiątą rocznicę śmierci. wiadomości botaniczne 21 (4): 221-224 (a review; in polish). – dermek a. 1976. huby lesov, polí a lúk. wiadomości botaniczne 21 (2): 144-145 (a review; in polish). bibliography 119 – grund d. w. & harrison k. a. 1976. nova scotian boletes. wiadomości botaniczne 21 (4): 271-272 (a review; in polish). – darimont p. 1973 recherches mycosociologique dans les forêts de haute belgique. kosmos ser. a 26 (4): 436-438. 1978. polish tulasnellales. 1. tulasnella inclusa (christ.) donk. acta mycol. 14 (1/2): 109-112. – grzyby wielkoowocnikowe rezerwatu lipówka w puszczy niepołomickiej. studia naturae ser. a. 17: 159-168. – černý a. 1976. lesnická fytopatologie. wiadomości botaniczne 22 (2): 134-135 (a review; in polish). – erhart j., erhartová m. & příhoda a. 1977. houby ve fotografii. wiadomości botaniczne 22 (2): 135-136 (a review; in polish). – dermek a. 1977. atlas našich hub. wiadomości botaniczne 22 (2): 136-137 (a review; in polish). – ulvinen t. et al. 1976. suursieniopas. wiadomości botaniczne 22 (3): 206-207 (a review; in polish). – blum j. 1976. les lactaires. wiadomości botaniczne 22 (3): 207-208 (a review; in polish). 1979. the geographical distribution of the tremellaceous fungi in poland. acta mycol. 15 (1): 75-144 (in polish with english summary). – macromycetes of the district of chrzanów and jaworzno. part 2. studia ośr. dokument. fizjogr. pan 7: 67-108 (in polish with english summary). – grzyby współżyjące czyli symbiotroficzne ryzobionty. (in:) k. zarzycki, zarys ekologii. (in:) s. białobok (ed.). brzozy betula l. nasze drzewa leśne 7: 283-284. państwowe wydawnictwo naukowe, poznań. – brodie h. j. 1975. the bird’s nest fungi. wiadomości botaniczne 23 (2): 135-136 (a review; in polish). – moser m. 1978. die röhrlinge und blätterpilze (polyporales, boletales, agaricales, russulales). wiadomości botaniczne 23 (2): 136-137 (a review; in polish). – cetto b. 1977. der große pilzführer. 1. wiadomości botaniczne 23 (3): 226-227 (a review; in polish). – michael e., hennig b., kreisel h. 1978. handbuch für pilzfreunde. 1. wiadomości botaniczne 23 (3): 227-228 (a review; in polish). – domański s. 1978. basidiomycetes (podstawczaki), aphyllophorales (bezblaszkowe), thelephorales (chropiatkowe). (in:) s. domański (ed.). mała flora grzybów 1 (3). wiadomości botanicz-botaniczne 23 (3): 229-229 (a review; in polish). 1980. materials on ecology of the polish tremellaceous fungi. acta mycol. 16 (1): 3-41 (in polish with english summary). – rattan s. s. 1977. the resupinate aphyllophorales of the north western himalayas. wiadomości botaniczne 24 (2): 169-170 (a review; in polish). – arnold g. r. et al. 1980. mykologisches wörterbuch. wiadomości botaniczne 24 (4): 294-295 (a review; in polish). – ryvarden l. 1976-1978. the polyporaceae of north europe. 1, 2. wiadomości botaniczne 24 (4): 295-296 (a review; in polish). – jordanov d., vanov s. g. & fakirova v. i. 1978. g’bite v b’łgarija. wiadomości botaniczne 24 (4): 296-297 (a review; in polish). – chang s.t. et al. 1978. the biology and cultivation of edible mushrooms. wiadomości botanicz-botaniczne 24 (4): 297-298 (a review; in polish). 1981. basidiomycetes (podstawczaki), tremellales (trzęsakowe), auriculariales (uszakowe), septobasidiales (czerwcogrzybowe). (in:) s. domański (ed.). mała flora grzybów 2: 1-408. państwowe wydawnictwo naukowe, warszawa-kraków. – taxonomy and systematic position of tremellaceous fungi. acta mycol. 16 (2): 159-193 (in polish with english summary). – macromycetes of the district of chrzanów and jaworzno. part 3. studia ośr. dokument. fizjogr. pan 8: 187-201 (in polish with english summary). – uwagi o grzybach wiekoowocnikowych rezerwatu łężczak koło raciborza. chrońmy przyrodę ojczystą 37 (2): 53-55. – dermek a., lizoň p. malý atlas hub. wszechświat 82 (10): 242-243 (a review; in polish). – cetto b. der große pilzführer. 2, 3. wszechświat 82 (10): 243 (a review; in polish). 120 z. heinrich and b. pleban 1982. uzupełnienie dotyczące mikoflory ojcowskiego parku narodowego. chrońmy przyrodę ojczystą 38 (1/2): 47-49. – klán j., vančura b. grzyby. wiadomości botaniczne 26 (1): 63-65 (a review; in polish). – kohlmeyer j., kohlmeyer e. marine mycology. the higher fungi. wiadomości botaniczne 26 (1): 65 (a review; in polish). – järva l., parmasto e. 1980. eesti seente koondnimestik. kosmos ser. a 31 (3/4): 255-256 (a review; in polish). 1983. (gumińska b., wojewoda w.) grzyby i ich oznaczanie (2nd ed.). państwowe wydawnictwo rolnicze i leśne, warszawa. – bolesław namysłowski (1882-1929) jako mikolog. w stulecie urodzin. wiadomości botaniczne 27 (3): 177-180. – problem pochodzenia wawelia regia namysłowski. wiadomości botaniczne 27 (4): 249-252. – grzyby towarzyszące jodle. (in:) a. jaworski, k. zarzycki, ekologia. (in:) s. białobok (ed.). jodła pospolita abies alba mill. nasze drzewa leśne 4: 414-417. państwowe wydawnictwo naukowe, warszawa–poznań. – jahn h. pilze an holzwachsen. kosmos 32 (1): 132-133 (a review; in polish). – dermek a. grzyby. wszechświat 84 (3): 75 (a review; in polish). – michael e., hennig b., kreisel h. handbuch für pilzfreunde. 4. wszechświat 84 (6): 150-151 (a review; in polish). – dähncke r. m. & dähncke s. m. 1980. 700 pilze in farbfotos. wiadomości botaniczne 27 (1): 83-84 (a review; in polish). – romagnesi h., gilles g. les rhodophylles des fôrets côtières du gabon et de la côte d’ivoire. wiadomości botaniczne 27 (4): 308-309 (a review; in polish). – jülich w., stalpers j. a. 1980. the resupinate non-poroid aphyllophorales of the temperate northern hemisphere. wiadomości botaniczne 27 (4): 309 (a review; in polish). 1984. polish tulasnellales. 2. tulasnella hyalina höhn. et litsch. acta mycol. 19 (1): 41-45. – bigelow h. e. 1982. north american species of clitocybe. part 1. wiadomości botaniczne 28 (2): 178-179 (a review; in polish). – michael e., hennig b., kreisel h. 1983. handbuch für pilzfreunde. 1. wiadomości botaniczne 28 (4): 321-322 (a review; in polish). – michael e., hennig b., kreisel h. 1983. hanbuch für pilzfreunde. 5. wiadomości botaniczne 28 (4): 322 (a review; in polish). – pegler d. n. 1977. a preliminary agaric flora of east africa. kosmos 33 (2): 225-227 (a review; in polish). 1985. (gumińska b., wojewoda w.) grzyby i ich oznaczanie (3rd ed.). państwowe wydawnictwo rolnicze i leśne, warszawa. – (stengl-rejthar a., wojewoda w.) expansion of the fungus clathrus archeri (berk.) dring (gasteromycetes) in europe and poland. zesz. nauk. uniw.jagiell. 752 prace bot. 13: 105-110. – baroni t. i. a revision of the genus rhodocybe maire (agaricales). wiadomości botaniczne 29 (3): 259-260 (a review; in polish). – horak e. entoloma (agaricales) in indomalaya and australasia. wiadomości botaniczne 29 (3): 260 (a review; in polish). – kotlaba f. zeměpisné rozšíření a ekologie chorošů (polyporales s. l.) v československu. kosmos 34(2): 340-341 (a review; in polish). 1986. (turowska i., podbielkowski z., wojewoda w.) rośliny zarodnikowe (3rd ed.). akademia medycz-medyczna, kraków. – polish dacrymycetales. 3. calocera glossoides (pers.) fr. fragm. flor. geobot. 29 (3/4): 461-464. – polish tulasnellales. 3. tulasnella violacea (johan-olsen ap. bref.) juel. acta mycol. 22(1): 99-102. – michael e., hennig b., kreisel h. hanbuch für pilzfreunde. 4. wiadomości botaniczne 31 (1): 92-93 (a review; in polish). – jülich w. 1984. die nichtblätterpilze, gallertpilze und bauchpilze, aphyllophorales, heterobasidiomycetes, gasteromycetes. (in:) kleine kryptogamenflora, band ii b/1, basidiomyceten, 1 teil. kosmos 34 (4): 668-670 (a review; in polish). – (wojewoda w., heinrich z., komorowska h.) macrobasidiomycetes new to the tatra national park (poland). acta mycol. 21 (1): 27-41. bibliography 121 – (wojewoda w., ławrynowicz m.) red list of threatened macrofungi in poland. (in:) k. zarzycki, w. wojewoda (eds). list of threatened plants in poland, pp.45-82. państwowe wydawnictwo naukowe, warszawa (in polish with english summary). 1987. (olesiński l., wojewoda w.) data on the macromycetes flora of the north-eastern poland. acta mycol. 21 (2): 193-232 (in polish with english summary). – mikologia w twórczości naukowej mariana raciborskiego. (in:) j. kornaś (ed.). marian raciborski. studia nad życiem i działalnością naukową. zesz. nauk. uniw. jagiell. varia 210: 59-78. – guzmán g. 1983. the genus psilocybe. wiadomości botaniczne 31 (1): 51(a review; in polish). – michael e., hennig b., kreisel h. hanbuch für pilzfreunde. 2. wiadomości botaniczne 31(1): 52-53 (a review; in polish). – svrček m., vančura b. grzyby środkowej europy. państwowe wydawnictwo rolnicze i leśne, warszawa (translation). – kreisel h. (ed.) pilzflora der deutschen demokratischen republik. basidiomycetes (gallert hutund bauchpilze). wiadomości botaniczne 31 (4): 242-243 (a review; in polish). 1988. (gumińska b., wojewoda w.) grzyby i ich oznaczanie (4th ed.). państwowe wydawnictwo rolnicze i leśne, warszawa. – (guzik j., wojewoda w.) morchella esculenta var. crassipes (ascomycotina, pezizales) in poland. fragm. flor. geobot. 33 (1/2): 211-215. – (kowalski s., wojewoda w., bartnik c., rupik a.) mycorrhizae of the birch and scotch pine and mycorrhizal fungi in cultivations contamined and free from industrial emissions. (in:) 2nd european symposium on mycorrhizae, abstracts, august 14–20, 1988: 58. prague. – kits van waveren e. the dutch, french and british species of psathyrella. kosmos 37 (1): 134-135 (a review; in polish). – (kreisel h., schauer f.) methoden des mykologischen laboratoriums. wiadomości botaniczne 32 (2): 129 (a review; in polish). 1989. (kowalski s., wojewoda w., bartnik c., rupik a.) mycorrhizal species composition and infection patterns in forest plantations exposed to different levels of industrial pollution. agric. ecosystems environ. 28: 249-255. – poprawki i uzupełnienia do książki grzyby środkowej europy. wszechświat 90 (9): 211-212. – eriksson j., hjortstam k., larsson k.-h., ryvarden l. the corticiaceae of north europe. 1-8. kosmos 38 (3): 399-400 (a review; in polish). – bondarceva m. a., parmasto e. h. clavis diagnostica fungorum urss. ordo aphyllophorales. fasc. 1. familiae hymenochaetaceae, lachnocladiaceae, coniophoraceae, schizophyllaceae. kosmos 38 (3): 400-401 (a review; in polish). – gilbertson r. l., ryvarden l. 1986-1987. north american polypores. 1-2. kosmos 38 (3): 402-403 (a review; in polish). 1990. heterobasidiomycetidae. (in:) k. zarzycki, u. korzeniak (eds). research progress report (1988-1990). polish bot. stud. guidebook ser. 1: 58. – grzyby związane z bukiem i jego zbiorowiskami. (in:) z. dzwonko, ekologia. (in:) s. białobok (ed.). buk – fagus sylvatica l. nasze drzewa leśne 10: 307-311. – (wojewoda w., heinrich z., komorowska h.) mycological investigations in the democratic people’s republic of korea. 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(in:) k. zarzycki, w. wojewoda, z. heinrich (eds). list of threatened plants in poland (2nd ed.), pp. 27-56. w. szafer institute of botany polish academy of sciences, kraków (in polish with english summary). – (wojewoda w., słomczyńscy a.p.) poradnik grzybiarza. prószyński i s-ka, warszawa. – (wojewoda w., słomczyńscy a.p.) podręczny atlas grzybów. państwowe wydawnictwo rolnicze i leśne, warszawa. – (zarzycki k., wojewoda w., heinrich z.) słowo wstępne. (in:) k. zarzycki, w. wojewoda, z. heinrich (eds). list of threatened plants in poland (2nd ed.), pp.3–5. w. szafer institute of botany polish academy of sciences, kraków (in polish with english summary). 1993. svrček m., vančura b. atlas grzybów. polska oficyna wydawnicza „bgw”, warszawa (translation). – larsen m. j., cobb-poulle l. a. phellinus (hymenochaetaceae). a survey of the world taxa. wiadomości botaniczne 37 (1/2): 202-203 (a review; in polish). – sunhede s. 1989. geastraceae (basidiomycotina). morphology, ecology, and systematics with special emphasis on north european species. wiadomości botaniczne 37 (1/2): 203-204 (a review; in polish). – tellería m. t. annotated list of the corticiaceae sensu lato (aphyllophorales, basidiomycotina), for peninsular spain and balearic islands. wiadomości botaniczne 37 (1/2): 204-205 (a review; in polish). – grochowski w. uboczna produkcja leśna. wiadomości botaniczne 37 (1/2): 204 (a review; in polish). – nordstein s. the genus crepidotus (basidiomycotina, agaricales) in norway. wiadomości bota-botaniczne 37 (1/2): 205 (a review; in polish). – tellería m. t. 1991. additions and corrections to the annotated list of the iberian corticiaceae (aphyllophorales, basidiomycotina). wiadomości botaniczne 37 (1/2): 204-205 (a review; in polish). – brandrud t. e., lindström h., marklund h., melot j., muskos s. cortinarius flora photographica. 1. wiadomości botaniczne 37 (1/2): 205-206 (a review; in polish). – senn-irlet b., jensen k. m., gulden g. arctic and alpine fungi. 3. wiadomości botaniczne 37 (1/2): 206 (a review; in polish). – breitenbach j., kränzlin f. fungi of switzerland. 1. ascomycetes. 2. non gilled fungi. 3. boletes and agarics. wiadomości botaniczne 37 (1/2): 206-207 (a review; in polish). – ryvarden l., gilbertson r. l. european polypores. 1. abortiporus lindtneria. wiadomości bota-botaniczne 37 (1/2): 207-208 (a review; in polish). – domański s. corticiaceae: kavinia rogersella; stephanosporaceae: lindtneria. (in:) s. domański (ed.). mała flora grzybów. 1 (6). wiadomości botaniczne 37 (1/2): 208 (a review; in polish). – żywa galareta. echa leśne 18 (1): 26. – fałszywa trufla. echa leśne 18 (2): 28. – jeden z pierwszych na wiosnę. echa leśne 18 (3): 28. – szkarłatna ozdoba. echa leśne 18 (4): 28. – siarkowożółty pasożyt. echa leśne 18 (5): 28. bibliography 123 – pułkownik grzybów. echa leśne 18 (6): 28. – piękny chociaż trujący. echa leśne 18 (7): 28. – nie może żyć bez osiki. echa leśne 18 (8): 28. – grzyb na grzybie. echa leśne 18 (9): 28. – sam się rozpływa. echa leśne 18 (10): 28. – grzyb jak ze szkła. echa leśne 18 (11): 30. – jeden z największych. echa leśne 18 (12): 28. 1994. profesor stanisław domański (1916-1993). acta mycol. 29 (1): 3-4. – stanisław domański (1916-1993). mycologist and phytopathologist. acta mycol. 29 (2): 131-140. – (wojewoda w., heinrich z., komorowska h.) macrofungi of north korea. wiadomości botanicz-botaniczne 37 (3-4): 125-128 (in polish with english summary). – (wojewoda w., słomczyńscy a.p.) podręczny atlas grzybów (2nd ed.). państwowe wydawnictwo rolnicze i leśne, warszawa. 1995. grzyby. (in:) a. medwecka-kornaś, s. loster, ojcowski park narodowy. (in:) z. mirek, j.j. wójcicki (eds). szata roślinna parków narodowych i rezerwatów polski południowej. przewodnik sesji terenowych 50. zjazdu ptb. polish bot. stud. guidebook series 12: 23-24. – grzyby. (in:) b. zemanek, k. towpasz, pogórze karpackie i bieszczady. (in:) z. mirek, j.j. wójcicki (eds). szata roślinna parków narodowych i rezerwatów polski południowej. przewodnik sesji terenowych 50. zjazdu ptb. polish bot. stud. guidebook series 12: 230–231. 1996. (kowalski s., rębisz a., wojewoda w.) badania stosunków biotycznych pomiędzy wybranymi grzybami mikoryzowymi i patogenicznymi. (in:) m. mańka (ed.). choroby roślin a środowisko, pp.51–57. polskie towarzystwo fitopatologiczne, poznań. – (kowalski s., obłoza e., wojewoda w.) susceptibility of ectomycorrhizal and ectendomycorrhizal fungi to ph of the environment. acta mycol. 31 (2): 127-136. – fungi of cracow during the years 1883–1994 with particular interest in macrofungi. studia ośr. dokument. fizjogr. 24: 75-111 (in polish with english summary). – grzyby wielkoowocnikowe. (in:) z. mirek, z. głowaciński, k. klimek, h. piękoś-mirkowa (eds). przyroda tatrzańskiego parku narodowego. tatry i podhale 3: 379-392. 1997. new taxa of fungi (aphyllophorales and agaricales) described by s. domański. acta mycol. 32 (2): 133-145. – grzyby z rodzaju płaskosz exobasidium, pasożyty wrzosowatych podawane z polski. (in:) k. zarzycki (ed.). rodzime rośliny wrzosowate w polskiej florze. arboretum bolestraszyce 5: 14–15. – życie i dzieło profesora stanisława domańskiego (1916-1993). wiadomości botaniczne 41 (1): 39-47. – mgr irena mazaraki (1912-1995). wiadomości botaniczne 41 (1): 39-47. – (wojewoda w., komorowska h.) notes on phleogena faginea (fungi, atractiellales). fragm. flor. geobot. 42 (1): 153-160. 1998. the macrofungi (basidiomycotina) of the beskid niski mts. (the polish carpathians). part i. heterobasidiomycetes and aphyllophorales. studia ośr. dokument. fizjogr. pan 25: 295-334 (in polish with english summary). – (mycological terms). (in:) z. otałęga (ed.). encyklopedia biologiczna 1, a-bn. wydawnictwo opres, kraków. – (mycological terms). (in:) z. otałęga (ed.). encyklopedia biologiczna 2, bo-dn. wydawnictwo opres, kraków. – (mycological terms). (in:) z. otałęga (ed.). encyklopedia biologiczna 3, do-gi. wydawnictwo opres, kraków. – (mycological terms). (in:) z. otałęga (ed.). encyklopedia biologiczna 4, gl-ja. wydawnictwo opres, kraków. – (mycological terms). (in:) z. otałęga (ed.). encyklopedia biologiczna 5, ją-kr. wydawnictwo opres, kraków. – (mycological terms). (in:) z. otałęga (ed.). encyklopedia biologiczna 6, ks-mn. wydawnictwo opres, kraków. – krytyczny wykaz grzybów podstawczaków (basidiomycetes) polski. działalność naukowa pan wybrane zagadnienia 6: 48-49. 124 z. heinrich and b. pleban – (wojewoda w., szymański j.) gene resources of selected cultivated mushroos growing wild in south-eastern poland. zeszyty problemowe postępów nauk rolniczych 463: 173-182 (in polish with english summary). 1999. red list of upper silesian macrofungi. centrum dziedzictwa przyrody górnego śląska raporty opinie 4: 8-51 (in polish with english summary). – polyporoid fungi of poland. state of examination and new systematic classification. zeszyty naukowe akademii rolniczej im. h. kołłątaja w krakowie 348 sesja naukowa 63: 47-56 (in polish with english summary). – preliminary characteristics of macromycetes in the magura national park. chrońmy przyrodę ojczystą 55 (1): 35-55 (in polish with english summary). – the corticioid fungi of the polish carpathians. wiadomości botaniczne 43 (3/4): 19-30 (in polish with english summary). – (mycological terms). (in:) z. otałęga (ed.). encyklopedia biologiczna 7, mo-oś. wydawnictwo opres, kraków. – (mycological terms). (in:) z. otałęga (ed.). encyklopedia biologiczna 8, ot-pr. wydawnictwo opres, kraków. – (mycological terms). (in:) z. otałęga (ed.). encyklopedia biologiczna 9, ps-si. wydawnictwo opres, kraków. – (wojewoda w., heinrich z., komorowska h.) new localities and new host for phleogena faginea (fungi, atractiellales) in poland. fragm. flor. geobot. ser. polonica 6: 199-202. – (wojewoda w., heinrich z., komorowska h.) macromycetes of oak-lime-hornbeam woods in the niepołomice forest near kraków (s poland) monitoring studies. acta mycol. 34 (2): 203-268. 2000. note from editor. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland. 1: 5-7. – asterodon ferruginosus pat. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 1: 9-12. – bondarzewia mesenterica (schaeff.) kreisel. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 1: 13-19. – eocronartium muscicola (pers.: fr.) fitzp. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 1: 25-28. – peniophora lilacea bourd. & galz. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 1:41-44. – porostereum spadiceum (pers.: fr.) hjortst. & ryv. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 1: 45-49. – stypella grilletii (boud.) p. roberts. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 1: 51-54. – xylobolus frustulatus (pers.: fr.) boid. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 1: 55-61. – typhula quisquiliaris (cantharellaes) a species new to poland. acta mycol. 35 (1): 29-35. – grzyby. (in:) j. staszkiewicz (ed.). przyroda popradzkiego parku krajobrazowego, pp.189-204. popradzki park krajobrazowy, stary sącz. – (mycological terms) (in:) z. otałęga (ed.). encyklopedia biologiczna 10, sj-ti. wydawnictwo opres, kraków. – (mycological terms) (in:) z. otałęga (ed.). encyklopedia biologiczna 11, tk-wr. wydawnictwo opres, kraków. – (mycological terms) (in:) z. otałęga (ed.). encyklopedia biologiczna 12, ws-ż-suplement. wydawnictwo opres, kraków. – (mycological terms) (in:) z. otałęga (ed.). encyklopedia biologiczna 13, suplement. taksonomiczny słownik polsko-łaciński i łacińsko-polski. wydawnictwo opres, kraków. – urbonas v. agarikiečiai (agaricales) 3 gijabudiečiai (entolomatales). (in:) v. urbonas et al. (eds). lietuvos grybai (mycota lithuaniae), viii. wiadomości botaniczne 44 (1/2): 101 (a review; in polish). – kuthan j., adamčiík s., terray j., antonín v. 1999. huby národného parku poloniny. wiadomości botaniczne 44 (3/4): 96-97 (a review; in polish). – (wojewoda w., kaźmierczakowa r.) clavaria zollingeri lév. – a new fungus to poland. chrońmy przyrodę ojczystą 55 (6): 7-12 (in polish with english summary). bibliography 125 2001. (szymański j., wojewoda w., kotlińska t.) genetic resources of selected cultivated mushrooms encountered in the wild in northern poland. (in:) w. święcicki, b. naganowska, b. wolko (eds). broad variation and precise characterization – limitation for the future. eucarpia, european association for research on plant breeding, section genetic resources pp. 108-111.institute of plant genetics, polish academy of sciences, poznań. – punctularia strigosozonata (fungi, corticiaceae) in poland and north korea. fragm. flor. geobot. 45 (1/2): 501-507. – new localities of rare and threatened species of geastrum (lycoperdales) in poland. acta mycol. 35 (2): 145-151. – m. lisiewska, m. ławrynowicz (eds). monitoring grzybów. wiadomości botaniczne 45 (1/2): 130-132 (a review; in polish). – j. banaszak, k. tobolski (eds). park narodowy bory tucholskie. wiadomości botaniczne 45 (1/2): 132-133. – kutorga e. 2000. ausuniečiai (pezizales). (in:) v. urbonas et al. (eds). lituvos grybai (mycota lithuaniae). iii, 5. wiadomości botaniczne 45 (1/2): 129-130 (a review; in polish). 2002. osiemdziesięciolecie urodzin i dwudziestolecie śmierci prof. dr hab. andrzeja nespiaka (1921-1981). wiadomości botaniczne 46 (3/4): 62-65. – mycological terms. (in:) m. karolczuk-kędzierska (ed.). encyklopedia powszechna. 1-8. wydawca r. kluszczyński, kraków. – dacryomyces ovisporus (dacryomycelales, basidiomycetes) new to the czech republik. czech mycol. 54 (1/2): 11-17. – publikacje w całości lub w części poświęcone prof. a. nespiakowi. wiadomości botaniczne 46 (3/4): 10-11. – macrofungi of the bolestraszyce arboretum near przeemyśl, se poland. arboretum bolestraszyce 9: 15-39 (in polish with english summary). – note from editor. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 5. w. szafer institute of botany , polish academy of sciences, kraków. – amylocorticium cebennense (bourdot) pouzar. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 7-9. w. szafer institute of botany, polish academy of sciences, kraków. – amylocorticium subincarnatum (peck) pouzar. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 11-13. w. szafer institute of botany, polish academy of sciences, kraków. – amylocorticium subsulphureum (p. karst.) pouzar. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 15-17. w. szafer institute of botany, polish academy of sciences, kraków. – bovista paludosa lév. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 23-26. w. szafer institute of botany, polish academy of sciences, kraków. – clavariadelphus truncatus (quél.) donk. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 27-30. w. szafer institute of botany, polish academy of sciences, kraków. – clavulicium macounii (burt) j. erikss. & boidin ex parmasto. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 31-34. w. szafer institute of botany, polish academy of sciences, kraków. – conohypha albocremea (höhn. & litsch.) jülich. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 35-37. w. szafer institute of botany, polish academy of sciences, kraków. – daedaleopsis tricolor (bull.: fr.) bondartsev & singer. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 39-44. w. szafer institute of botany, polish academy of sciences, kraków. – fomitopsis hippophaeicola (h. jahn) fiasson & niemelä(in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 55-59. w. szafer institute of botany, polish academy of sciences, kraków. – irpicodon pendulus (alb. & schwein.: fr.) pouzar. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 77-79. w. szafer institute of botany, polish academy of sciences, kraków. 126 z. heinrich and b. pleban – punctularia strigosozonata (schwein.) p. h. b. talbot. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 81-83. w. szafer institute of botany, polish academy of sciences, kraków. – scotomyces subviolaceus (peck) jülich. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 99-101. w. szafer institute of botany, polish academy of sciences, kraków. – syzygospora pallida hauerslev. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 111-113. w. szafer institute of botany, polish academy of sciences, kraków. – thanatephorus sterigmaticus (bourdot) p. h. b. talbot. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 115-117. w. szafer institute of botany, polish academy of sciences, kraków. – tubulicrinis borealis j. erikss. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 127-129. w. szafer institute of botany, polish academy of sciences, kraków. – (wojewoda w., heinrich z., komorowska h.) trichaptum biforme (fr.) ryvarden. (in:) w. woje-wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 119-126. w. szafer institute of botany, polish academy of sciences, kraków. – (wojewoda w., komorowska h., piątek m.) hymenochaete cruenta (pers.: fr.) donk. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 2: 69-76. w. szafer institute of botany, polish academy of sciences, kraków. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity in poland, vol. 7. w. szafer institute of botany, polish academy of sciences, kraków. – morphology of some rare and threatened polish basidiomycota. acta mycol. 38 (1-2): 3-20. 2004. (wojewoda w., heinrich z., komorowska h.) macrofungi of the reserve „bór na czerwonen” in the orawa-nowy targ basin. fragm. flor. geobot. polonica 11: 177-189 (in polish with english summary). – (wojewoda w., heinrich z., komorowska h.) macrofungi in north korea. działalność naukowa pan 18: 61-62 (in polish with english summary). 2005. grzyby krzemionek podgórskich. (in:) m. szczepańska, e. pilecka (eds). geologiczno-przy rod nicze rozpoznanie terenów pogórniczych krzemionek podgórskich dla potrzeb ochrony ich wartości naukowo-dydaktycznych i ekologicznych. wydawnictwo instytutu gospodarki surowcami mi neral nymi i energią pan, kraków, pp.75-87, 127-130. – botryobasidium laeve (j. erikss.) parmasto. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 3: 17-21. w. szafer institute of botany, polish academy of sciences, kraków. – coniophora olivacea (pers.: fr.) p. karst.. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 3: 27-30. w. szafer institute of botany, polish academy of sciences, kraków. – helicobasidium purpureum pat. in: w. wojewoda (ed.) atlas of the geographisal distribution of fungi in poland 3: 39-42. w. szafer institute of botany, polish academy of sciences, kraków. – leucogyrophana olivascens (berk. & m. a. curtis) ginns & weres. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 3: 55-57. w. szafer institute of botany, polish academy of sciences, kraków. – lycoperdon mammiforme pers.: pers. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 3: 59-63. w. szafer institute of botany, polish academy of sciences, kraków. – oligoporus obductus (berk.) gilb. & ryvarden. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 3: 81-84. w. szafer institute of botany, polish academy of sciences, kraków. – onygena equina (willdenow) pers.: fr. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 3: 85-88. w. szafer institute of botany, polish academy of sciences, kraków. – veluticeps ambigua (peck) hjortstam & telleria. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 3: 115-117. w. szafer institute of botany, polish academy of sciences, kraków. bibliography 127 – (wojewoda w., karasiński d.) clavaria zollingeri lév. (in:) w. wojewoda (ed.). atlas of the geographical distribution of fungi in poland 3: 23-26. w. szafer institute of botany, polish academy of sciences, kraków. 2006. auriculariopsis albomellea (agaricales, schizophyllaceae) new for poland. acta mycol. 41 (1): 49-54. – prof. dr hab. inż. bronisław zyska (1924-2006). wiadomości botaniczne 50 (3/4): 32-37. – (wojewoda w., ławrynowicz m.) red list of the macrofungi in poland. (in:) z. mirek, k. zarzycki, w. wojewoda w., z. szeląg (eds). red list of plants and fungi in poland, pp. 55-70. w. szafer institute of botany, polish academy of sciences, kraków. 2007. (wojewoda w., wojewoda m.) grzyby inwazyjne: okratek australijski clathrus archeri i pierścieniak uprawny stropharia rugosoannulata – w beskidzie wyspowym. wszechświat 108 (10/12): 300-303. 2008. macrofungi of the ojców national park. (in:) a. klasa, j. partyka (eds). monografia ojcowskiego parku narodowego. przyroda: 317-334. ojcowski park narodowy, ojców (in polish with english summary). 2009. (wojewoda w., karasiński d.) p. łakomy, h. kwaśna 2008. atlas hub. multico oficyna wydawnicza, warszawa, 2008, 172 str., wielobarwne fot. 274. wiadomości botaniczne 53 (3/4): 196-197 (a review; in polish). 2010. rozstania. mgr mieczysław aleksander mazaraki (1913-2003) – niezwykły nauczyciel, botanik, zoolog, historyk, twórca muzeum. wiadomości botaniczne 54 (3/4): 51-58. – professor barbara gumińska. her life and achievements. acta mycol. 45 (1): 3-10. – laricifomes officinalis in the gorce mountains (s poland). acta mycol. 45 (2): 129-131. – (wojewoda w., karasiński d.) invasive macrofungi (ascomycota and basidiomycota) in poland. (in:) z. mirek (ed.). biological invasions in poland 1: 7-21. w. szafer institute of botany, polish academy of sciences, kraków. – (wojewoda w., mleczko p.) bibliography of publications by barbara gumińska. acta mycol. 45 (1): 11-16. zofia heinrich & barbara pleban 2014-01-01t11:52:12+0100 polish botanical society hymenoscyphus subcarneus, a little known bryicolous discomycete found in the białowieża national park hans otto baral1 and lothar krieglsteiner2 1blaihofstr. 42, d 72074 tübingen 2konrad adenauer str. 32, d 73529 schwäbisch gmünd b a r a l h . o . , k r i e g l s t e i n e r l .: hymenoscyphus subcarneus, a little known bryicolous discomycete found in the białowieża national park. acta mycol. 41 (1): 11 20, 2006. the discomycete hymenoscyphus subcarneus was found to grow parasitic on the liverwort cephalozia catenulata in the białowieża national park (poland), and is described and illustrated from the fresh collection. two characters, the ascus apical ring structure of the calycina type and the contents of the living paraphyses (multiguttulate by low refractive vacuolar bodies), have not been reported previously. in addition, a dried collection on pohlia gracilis from switzerland, grimsel area, was studied. the relationship, taxonomy and infraspecific variation of the fungus are discussed. a new genus, roseodiscus, is established to accomodate this bryicolous fungus together with two very similar equiseticolous species, hymenoscyphus rhodoleucus and h. equisetinus. the three species are macroscopically characterized in the fresh state by a pale rosaceous lilaceous hymenium and a mostly slender, concolorous or whitish stipe. roseodiscus resembles hymenoscyphus in various respects, but sharply deviates in the apical ring type which appears to indicate a more close relationship with genera like calycina or stamnaria. rhizoscyphus ericae which forms a mycorrhiza with roots of ericaceae, is compared with roseodiscus. based on vital observations of r. ericae this species is believed to be congeneric with the type of pezoloma, p. griseum. key words: roseodiscus, rhizoscyphus ericae, hymenoscyphus rhodoleucus, bryicolous introduction in june, 1996, the second author and angelika huber, the latter working on bryophytes, had the opportunity to collect in the nature reserve of białowieża national park. the visit in poland came about by an exchange between the universities of regensburg and łódź. an interesting discomycete could be found that is described and illustrated here. acta mycologica vol. 41 (1): 11-20 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 12 h. o. baral and l. krieglsteiner description of species hymenoscyphus subcarneus (sacc.) o. kuntze, rev. gen. pl. 3 (2): 486 (1889) nom. illeg. [non hymenoscyphus subcarneus (schumach.) j. schröt., schles. kryptfl. 3.2: 69 (1893) ≡ phaeohelotium subcarneum (schumach.) dennis, kew bull. 25: 355 (1971)] [≡peziza subcarnea cooke and peck in cooke, bull. buffalo soc. nat. sci. 2: 295 (1875), nom. illegit.; non peziza subcarnea schumach., flora danica 12, tab. 2084, fig. 1 (1832)] ≡ phialea subcarnea sacc., syll. fung. 8: 265 (1889) ≡helotium destructor peck in white, mycologia 34: 163 (1942) [nom. nov., non helotium subcarneum (schumach.) sacc., michelia 2(7): 260 (1881)] description of the sample from poland (* living state) apothecia scattered to subgregarious, fresh (0.4-)0.5-1.3(-1.5) mm, shallowly cupto saucer-shaped, whitish but with a distinct pale pink tint, stalk 0.6-1.2 x 0.20.35 mm, abruptly delimited from cup, concolorous, pale ochraceous towards base, exterior nearly smooth under a hand lens, at higher magnification delicately pubescent, especially near margin, margin of large apothecia often undulating to somewhat irregular. ectal excipulum of a horizontally (10-20°) oriented hyaline textura prismatica of thin-walled cells, individual cells *(12-)20-45(-67) x (6-)10-13(-16) μm in both surface view and median section, externally covered by a loose network of 3-5 μm wide eguttulate hyphae, towards margin of t. porrecta oriented at c. 10°, cells (in koh) 13-18 x 2.5-4 μm, not differentiated from covering hyphae which terminate here in somewhat projecting hair-like ends that contain in the living state low-refractive guttules (vbs), in stalk of t. porrecta. no crystals and no amyloid or dextrinoid tissue observed. medullary excipulum near ectal excipulum of a broad textura porrecta, in centre of a small region of dense t. intricata (individual cells here in koh 12-25 x 3-4 μm). asci *52-66 x 6.5-7.5(-10) μm, in koh 42-57 x (5.5-)6-7(-7.5) μm, pars sporifera *(15-)20-26 μm long, 8-spored, apex strongly conical, apical ring deep blue in iki (type bb euamyloid), of the calycina-type, 0.6-1 μm high and 0.9-1.1 μm wide (dead state), base attenuated or rather broad, arising from croziers. spores biseriate within the living asci, free *(5.2-)6-8.5(-10) x (1.8-)2-2.6(-3) μm, in koh c. 6-8 x 2-2.3 μm, of variable length, slightly to mainly strongly clavate or pyriform or even somewhat cuneate (triangular), hyaline, smooth, with a few very minute lbs in each spore half, no surrounding sheath observed. paraphyses cylindrical to slightly clavate, even in dead state 3-7 μm shorter than the asci, terminal cell *(11-)16-37 x 2.8-3.5(-4) μm, in living state multiguttulate in the upper 25-40 μm by globose, low (to medium) refractive vbs (large drops sometimes strongly elongated), vbs disappearing in koh, invisible in dead cells. examined collection: poland, białowieża national park, ca. 1 km north of administration centre białowieża, ca. 130 m a.s.l., parasitic on living, green gametophytes of cephalozia catenulata (hüb.) lindb. growing on a log of picea abies lying on the ground on wayside, 28.vi.1996, l. krieglsteiner and a. huber (figs 1-2; 3). specimen deposited in the herbarium krieglsteiner (stuttgart) and private herbarium h.-o. baral (h.b. 5531). hymenoscyphus subcarneus, a little known bryicolous discomycete 13 further collection studied: switzerland, bern, grimsel-guttannen area, bächlisboden, sander, 2170 m a.s.l., over granite, parasitic on pohlia gracilis, 26.viii.1986, b. senn-irlet, b.i. 86/88, h.b. 3341. reports of hymenoscyphus subcarneus in the literature a d e (1935: 24) reported under the name phialea subcarnea two collections from austria (tirol, vennatal and roßkogel above inzingeralpen, summer 1925 and 1927). his description fits more or less to ours, except that he states the spores to be “elongate, often slightly curved, with narrow, bright mucilaginous sheath”. the “sheath” might be a mistake for dead spores with detached plasma, perhaps observed from within the asci. ade gave the spore size as “6-7 x 2-5” μm, but in view of the indicated ascus size (40-50 x 5-6 μm) and the irregularly biseriate spore arrangement, ade´s spore width is probably an error for 2-2.5 μm. the rather narrow excipular cells (“upto 18.5 x 3.5 μm”) forming a t. porrecta might be due to the immature collection. w h i t e (1942, as helotium desctructor) examined much over a dozen of collections from u.s.a. (new york, michigan) and canada (alberta, new hampshire, ontario) including the type specimen (u.s.a., new york, indian lake, on jungermannia sp., c. h. peck 319). the apothecia are distinctly smaller than in our collection: 0.2-0.4(-0.5) mm diam., total height upto 1 mm, “scarcely visible to the unaided eye” (possibly measured in the dry state). ascus (40-50 x 5-6 μm) and spore size (4.56 x 2-2.5 μm) are a bit smaller than in our collection when comparing the dead state. the pyriform spore shape as well as the conical ascus apex and presence of croziers as depicted by white perfectly fit to our collection. d e n n i s (1964: 68, as hymenoscyphus subcarneus) studied the excipular characters from the type. in concordance with our fungus, dennis reported the cells to be rather broad, “about 20-30 x 10 μm, lying at a very low angle to the surface”. with little doubt, l i z o ň (1992: 48) described under the name h. subcarneus the same species. the material studied by him was collected in slovakia (belanské tatry, in valle holubyho dolina pr. tatranská kotlina, 10.viii.1956, m. svrček, prm 817510). both the ascus width (40.5-60 x 5-6 μm) and the spore length (4.5-6 x 2-2.5 μm) given by lizoň (gained from dead herbarium material), appear to have partly been copied from white, since lizoň stated that he could examine “only young, not fully mature fruit-bodies”. tr i e b e l (1999: 10) reported a recent collection from canada (ca. 75 km nne of québec, forêt montmorency). the fungus was identified by p. döbbeler and said to grow on at least five different bryophytes. except for the apothecial colour (distinctly pink) no further comments were published. the here studied collection from switzerland (on pohlia gracilis) differs from all the above in distinctly longer asci (80-100 x 6-9 μm, living state) and ascospores (ellipsoid-oblong, *8.5-13.5 x 2.7-3.2 μm). the question whether or not this represents a separate species must await further field work. similar variability was observed by us in hymenoscyphus rhodoleucus (fr.: fr.) w. phillips and h. equisetinus (velen.) dennis, two species which are obviously closely related to h. subcarneus (see below). due to a rather high variation in spore size, the delimitation between these two equiseticolous taxa is still not fully settled. 14 h. o. baral and l. krieglsteiner homonymy with peziza subcarnea schumach. a somewhat similar species with a similar name, phaeohelotium subcarneum (schumach.) dennis, has also pinkish apothecia but lacks a stalk and was mainly reported from bark and wood. it was redescribed by h ö h n e l (1926: 99), g r e l e t (1948: 108), d e n n i s (1971: 355), b r e i t e n b a c h & k r ä n z l i n (1981, pl. 173), and b a r a l (1986: 17) from non-authentic material. the species was even combined in hymenoscyphus gray as h. subcarneus (schumach.) j. schröt., four years after o. kuntze transferred peziza subcarnea cooke and peck to that genus. d e n n i s (1971) is obviously in error when citing the species as phaeohelotium subcarneum (schumach. ex sacc.) dennis, because peziza subcarnea cooke and peck is illegitimate, not the earlier peziza subcarnea schumach. the few known reports of phaeohelotium subcarneum are possibly not homogeneous, at least the brief descriptions support such a conclusion. that p. subcarneum is a synonym of hymenoscyphus imberbis (bull.: fr.) dennis was erroneously cited by ve r k l e y (1995: 180). in fact, baral (b a r a l & k r i e g l s t e i n e r 1985: 129) stated that p. subcarneum in the sense of breitenbach and kränzlin seems to belong to h. imberbis or a closely related species. p. subcarneum s. b a r a l (1986) has a calycinatype of apical ring and is therefore unrelated to hymenoscyphus (incl. phaeohelotium kanouse). the ectal excipulum of t. globulosa without a covering layer appears to separate it sharply at generic level from hymenoscyphus subcarneus (sacc.) o. kuntze. furthermore, the apothecia are fresh greyish-white and turn only pinkish with age or when dried. the generic position of this species remains unclear upto now. generic position of hymenoscyphus subcarneus (sacc.) o. kuntze d e n n i s (1964: 43) and l i z o ň (1992) believed that the bryicolous species is a normal hymenoscyphus. however, the rather wide generic concepts of the two authors included in hymenoscyphus also species of calycina (nees) gray, due to the frequent neglection of apical ring types in light-microscopical studies. the apical ring of h. subcarneus is of the calycina-type: rather thick in optical section, widest in its upper part which extends to the very apex, here most strongly reactive (b a r a l 1987, fig. 26; ve r k l e y 1995: figs. 4.74-76, 186, “type viii: chlorociboria-pezizella-calycina”), and the ascus apex is strongly conical. this type of ascus apex is very much unlike that of typical members of hymenoscyphus with a truncate, broadly conical apex and a narrow ring that is usually only present in the lower half of the apical thickening as two very thin amyloid lines which are not widened apically (b a r a l 1987, figs 10-12). the guttulate paraphyses and the presence of a covering layer of narrow hyphae over the excipulum would, however, fit very well in that genus. the genera bryoscyphus spooner and muscicola (velen.) svrček likewise differ from our fungus in ascus structure (hymenoscyphus-type; b a r a l & m a r s o n 2005), the latter also in the presence of distinct, strongly uncinate hairs. two taxa growing on stems of equisetum, hymenoscyphus rhodoleucus and h. equisetinus, strongly resemble our species in their pinkish apothecia, but also in their microscopical characters. like h. subcarneus, both have apical rings of the caly hymenoscyphus subcarneus, a little known bryicolous discomycete 15 cina-type (b a r a l & m a r s o n 2005) and are therefore clearly misplaced in the genus hymenoscyphus. s v r č e k (1959) already compared one of the two (as phialea rhodoleuca) with our species (as phialea subcarnea). placement of h. rhodoleucus in phialea (fr.) gillet ( cyathicula de not.) was favoured by n a n n f e l d t (1932) while d e n n i s (1956, 1978) preferred to assign the two equiseticolous species to hymenoscyphus. macroscopically, our bryicolous species indeed somewhat resembles a cyathicula. however, members of that genus have a strongly gelatinized excipulum and often also crystals on the exterior, both absent in our species. the ascus structure varies strongly among the accepted species (tr i e b e l & b a r a l 1997 as crocicreas fr.), comprising both the hymenoscyphusand the calycina-type. however, the latter occurs only in the type species of crocicreas which should better be distinguished at the generic level. the guttulate paraphyses and the covering layer in our species would fit to cyathicula. species of calycina differ from our fungus in having a more gelatinized ectal excipulum of much shorter cells which often lie at a higher angle to the surface, and in constantly lacking an external covering hyphal layer. the hairs of calycina emerge directly from the prismatic excipular cells. furthermore, spores of calycina are usually homopolar. last but not least, the vbs in the living paraphyses of calycina species are generally strongly elongate, not multiguttulate. the genus calycina was transferred form the helotiaceae to the hyaloscyphaceae by the first author (b a r a l & k r i e g l s t e i n e r 1985). the genus stamnaria fuckel is characterized by a gelatinized layer outside the ectal excipulum, and the presence of carotenoids in both excipulum and paraphyses. otherwise it shows some similarities with our species, including the apical ring which is of the calycina-type in s. equiseti (hoffm.) sacc. (k ü n k e l e et al. 2005). the genus rhizoscyphus w. y. zhuang and korf was described in z h a n g & z h u a n g (2004) to accomodate r. ericae (d. j. read) w. y. zhuang and korf (≡ hymenoscyphus ericae (d. j. read) korf and kernan), a species with a mycorrhizal connection to roots of ericaceae. the erection of a new genus was mainly based on molecular data, but also on the symbiontic relationship with the host plants. huhtinen (h a m b l e t o n et al. 1999) redescribed the species in detail including vital characters. following his report, the living paraphyses are without any refractive vacuoles and the apical ring is of the calycina-type. characteristic of the species is the hyphoid margin which gives the apothecia a hairy appearance, also the sometimes furcate apices of paraphyses and the simple-septate ascus bases with often a downward protuberance. clearly, the apical ring excludes hymenoscyphus s.str. the microscopical data are indeed similar to h. subcarneus, but the subsessile hairy apothecia more resemble a hyaloscyphaceae. recently the first author had the opportunity to study a fresh specimen of r. ericae detected by p. zinth. the apothecia were formed on the rootlets of a rhododendron sp. that grows in an semi-sterilized artificial medium in a pot culture in his house in egelsbach near frankfurt (hessen). the specimen fully matches huhtinen´s analysis. in addition, the mature living asci each contained one globose, refractive drop (4-4.5 μm diam.) below the pars sporifera similar as in the genus psilachnum höhn. these drops disappear in dead asci and were also seen in fresh finds of pezoloma ciliifera (p. karst.) korf and p. marchantiae (sommerf.) benkert 16 h. o. baral and l. krieglsteiner (b a r a l & m a r s o n 2005), two species which share some further characteristics of r. ericae like the subsessile apothecia, the type of apical ring and the eguttulate, sometimes apically furcate paraphyses. the two species differ in an external gel layer and in marginal teeth composed of narrow agglutinated hyphae. these teeth are a striking feature of the typical species of pezoloma clem., but are absent in many other taxa assigned to that genus. the hyphae of the teeth indeed resemble the hair-like protrusions of r. ericae. similar differences between dentate and adentate apothecia are accepted within the genus cyathicula. however, the genus pezoloma in the current circumscription (e.g., g a r c i a & va n vo o r e n 2005) is heterogenous and includes also taxa with a typical hymenoscyphus-type of apical ring. the presence of an external gel layer which is used to define the genus is most probably a polyphyletic feature whereas differences in the apical ring type appear to have much more been conserved during evolution. based on the above data it seems wise to restrict the genus pezoloma to species with a calycina-type of apical ring. the morphology of r. ericae appears to be quite close to p. ciliifera and p. marchantiae. these in turn are surely very closely related to the type species of pezoloma, p. griseum clem., which has also a toothed margin. while the former two species are partly found growing on mosses (sphagnum, marchantia), p. griseum was reported by c l e m e n t s (1911) to grow on rootlets of betula, suggesting a mycorrhizal life style similar as in r. ericae. moreover, r. ericae was two times isolated as endophytic mycelium in a liverwort (cephaloziella, c h a m b e r s et al. 1999). it is therefore believed that the type species of rhizoscyphus and pezoloma belong to the same genus. hence the following new combination is proposed. molecular research on species of pezoloma and a comparison with the sequence of r. ericae would highly be appreciated. pezoloma ericae (d. j. read) baral comb. nov. basionym: pezizella ericae d. j. read, trans. brit. mycol. soc. 63: 381 (1974) the genus moserella pöder and scheuer (1994), with the single species m. radicicola pöder and scheuer, was described as having very minute, strongly convex (capitate), white (to yellowish) apothecia with long and very slender stalks growing hypogeus on ectomycorrhizal rootlets of picea abies. the ascus apex is very precisely illustrated, clearly showing the calycina-type of apical ring. the microscopical features are not unlike our fungus, except for the spores which are said to be finely warted. possibly moserella is also closely related to pezoloma. there appears to exist no genus to which the three species with rose apothecia can safely be assigned. a new genus is therefore proposed. roseodiscus baral gen. nov. speciis generis hymenoscyphus sensu stricto similis sed asci apice valde conici, annulus apicalis typo calycina, apothecia hymenio pallide roseo-lilacea, excipulum ectale textura prismatica, pars externa hyphis tecta. in equisetis vel bryophyta crescunt. apothecia 0.5-1.8 mm diam., disc flat, pale rose-lilaceous (salmon-coloured to carneous or pinkish), stipe whitish to concolorous, rather translucent, 0.25-2 x 0.20.6 mm, base hyaline to faintly brownish, superficial or erumpent from beneath epidermis, whole exterior partly very finely pubescent. ectal excipulum of large-celled, thin-walled, non-gelatinized textura prismatica oriented at a low angle (10-20°) to hymenoscyphus subcarneus, a little known bryicolous discomycete 17 the surface, externally covered by a loose network of hyphae from which hair-like projections may emerge, towards margin of t. porrecta. medullary excipulum nongelatinized, of dense t. intricata in centre, on flanks near ectal excipulum forming a broad t. porrecta. asci 8-spored, with strongly conical apex with deeply amyloid (bb, rb) apical ring of calycina-type, arising from croziers. paraphyses *(2.5-)33.5(-4) μm wide, containing either non-refractive vacuoles (species on equisetum) or near apex multiguttulate by slightly (to medium) refractive vacuolar bodies (vbs). ascospores cylindric-clavate to narrowly ellipsoid, partly slightly constricted in the middle, aseptate (rarely a few septate) when within the living asci, overmature often 1-septate, with a low lipid content, containing glycogen bodies. in bogs and swamps, parasitic to saprotrophic on bryophyta and equisetum. typus generis: roseodiscus rhodoleucus (fr.: fr.) baral, comb. nov. basionym: peziza rhodoleuca fr., obs. mycol. ii: 306 (1818) ≡ hymenoscyphus rhodoleucus (fr.: fr.) w. phillips, brit. discom. p. 131 (1887) further species included: roseodiscus equisetinus (velen.) baral, comb. nov. basionym: septatium equisetinum velen., monogr. discom. bohem. p. 212 (1934) ≡ hymenoscyphus equisetinus (velen.) dennis, brit. ascom. p. 138 (1978) roseodiscus subcarneus (sacc.) baral, comb. nov. basionym: phialea subcarnea sacc., syll. fung. 8: 265 (1889) roseodiscus resembles the genus hymenoscyphus by macroscopy, but sharply differs in the calycina-type of amyloid apical ring (see above). in one collection of r. rhodoleucus an associated phialidic anamorph could be observed: broadly lageniform hyaline phialides (*10.5-17 x 4-5.7 μm) with a narrow collarette produced cylindric-ellipsoid, basally truncate phialoconidia (*6.5-10 x 1.6-3 μm). the same phialides were also sometimes seen emerging from the ectal excipulum. the new genus is obviously more related to stamnaria, pezoloma and calycina than to hymenoscyphus. this is confirmed in the phylogenetic analysis of z h a n g & z h u a n g (2004) where h. rhodoleucus is found far away from the main part of hymenoscyphus, but clustering together with calycina herbarum (pers.) gray (hyaloscyphaceae), whereas pezoloma ericae is found in a separate clade quite far away from these two. ecology and parasitism r. subcarneus appears to be a montaneous to subalpine and probably also boreal species, at least in europe. a d e ´s (1935) collections were reported to have grown in the alps between 1500 and 1800 m on north-exposed slopes, the one of s v r č e k (1959) in c. 1200 m. in north america the species was collected, e.g., in the adirondack mountains (eastern u.s.a.) and in canada. w h i t e (1942) assumed that the species is not uncommon in that area. the n-american collections were made in (august-)september, except for peck´s type (july), while those from europe were made during june-august. the reported host plants are: blepharostoma trichophyllum dum. (ade), cephalozia catenulata (present study), dicranum flagellare hedw. (white, ontario), junger18 h. o. baral and l. krieglsteiner mannia sp. (peck´s type, new york), “foliose hepatic” (svrček, lizoň), various bryophytes like dicranum sp., hypnum sp., lepidozia reptans (l.) dum., polytrichum sp., tetraphis pellucida hedw. (triebel). according to white, “there seem to be no morphological differences between the forms on liverworts and those on mosses”. the mosses grew on old trunks (usa, canada, poland), or over stony ground (“glimmerschiefer”, tirol). r. subcarneus appears to be a parasitic species which has already been stated by peck and h. s. jackson (w h i t e 1942: 165), a d e (1935), and s v r č e k (1959). h. s. jackson stated that “the apothecia are best located by first searching out the brown areas in green patches of mosses where the plants have evidently been killed by the fungus”. also ade described a parasitic growth in his collections. the fungus is said to kill the tissue and thereby changes the colour of the moss population by forming circular yellow-brown lesions (a d e 1935). more or less distinct lesions could also be observed in our specimen from poland. acknowledgements. we want to thank a. bresinsky (d regensburg) and m. ławrynowicz (łódź) for the possibility to drive to poland, angelika huber (d münchen) for the company at the visit and the excursions as well as for the determination of cephalozia catenulata. we are also much indebted in b. and g. sumorok for the enormous logistic help. thanks also to j. b. faliński the director of geobotanical station (university of warsaw) for the licence for collecting permission in the national park. p. döbbeler (d münchen) and c. hahn (d augsburg) contributed to the correct determination of the fungus by calling into our attention the paper of p. lizoň. s. huhtinen is thanked for valuable comments on the nomenclature and z. pouzar for reviewing the latin diagnosis. references a d e a. 1935. bemerkenswerte pilze. kryptog. forschungen 2: 23 27. b a r a l h. o. 1986. beilage zum beiheft 6. z. mykol., typoskript., 19 pp. b a r a l h. o. 1987. der apikalapparat der helotiales. eine lichtmikroskopische studie über arten mit amyloidring. z. mykol. 53 (1): 119 136. b a r a l h. o., m a r s o n g. 2005. in vivo veritas. over 10.000 scans of fungi and plants, with materials on vital taxonomy and xerotolerance (dvd rom). b a r a l h. o., k r i e g l s t e i n e r g. j. 1985. bausteine zu einer askomyzeten flora der bundesrepublik deutschland: in süddeutschland gefundene inoperkulate diskomyzeten mit taxonomischen, ökolo gischen, chorologischen hinweisen und einer farbtafel. z. mykol., beiheft 6: 1 160. b r e i t e n b a c h j., k r ä n z l i n f. 1981. pilze der schweiz, ascomyzeten. luzern. c h a m b e r s s. m., w i l l i a m s p. g., s e p p e l t r. d., c a i r n e y j. w. g. 1999. molecular identification of hymenoscyphus sp. from rhizoids of the leafy liverwort cephaloziella exiliflora in australia and antarctica. mycol. res. 103 (3): 286 288. c l e m e n t s f. e. 1911. nova fungorum coloradensium genera. minnesota botanical studies 4 (2): 185 188. d e n n i s r. w. g. 1956. a revision of the british helotiaceae in the herbarium of the royal botanic garden, with notes on related european species. mycol. pap. 62: 1 216. d e n n i s r. w. g. 1964. remarks on the genus hymenoscyphus, with observation on sundry species re ferred by saccardo and others to the genus helotium, pezizella or phialea. persoonia 3: 29 80. d e n n i s r. w. g. 1971. new or interesting british microfungi. kew bull. 25: 335 374. g a r c i a g., va n vo o r e n v. 2005. un discomycète inoperculé plutôt discret, pezoloma ciliifrea, et remarques sur le genre pezoloma. bull. mens. soc. linn. lyon 74, num. spécial, p. 115 129. g r e l e t l. j. 1948. les discomycètes de france 18. rev. myc. (paris) n. s. 13: 105 134. h a m b l e t o n s., h u h t i n e n s., c u r r a h r.s. 1999. hymenoscyphus ericae: a new record from western canada. mycol. res. 103: 1391 1397. h ö h n e l f.v. 1926. über die gattung pezizella fuckel. 2. mitteilung. mitt. bot. lab. techn. hochsch. wien 3: 94 108. hymenoscyphus subcarneus, a little known bryicolous discomycete 19 k ü n k e l e u., l o h m e y e r t. r., b a r a l h. o. 2005. stamnaria americana, ein in auwäldern vermut lich häufiger, aber aus deutschland bisher nicht berichteter parasit an equisetum hyemale. mycol. bav. 7: 3 20. l i z o ň p. 1992. the genus hymenoscyphus (helotiales) in slovakia, czechoslovakia. mycotaxon 45: 1 59. n a n n f e l d t a. 1932. studien über die morphologie und systematik der nicht lichenisierten inoperculaten discomyzeten. nova acta regiae soc. scient. upsal., ser. 4, 8: 1 368. p ö d e r r., s c h e u e r c. 1994. moserella radicicola gen. et sp. nov., a new hypogeous species of leotiales on ectomycorrhizas of picea abies. mycol. res. 98 (11): 1334 1338. s v r č e k m. 1959. new ascomycetes for čzechoslovakia (in čzech). česka mykol. 13: 124 125. tr i e b e l d., b a r a l h. o. 1996. notes on the ascus types in crocicreas with a characterization of se lected taxa. sendtnera 3: 199 218. tr i e b e l d., b a r a l h. o. 1999. microfungi exsiccati fasc. 15 18. ve r k l e y g. j. m. 1995. the ascus apical apparatus in leotiales: an evaluation of ultrastructural char acters as phylogenetic markers in the families sclerotiniaceae, leotiaceae, and geoglossaceae. proef schrift, leiden, 209 pp. w h i t e w. l. 1942. studies in the genus helotium i. a review of the species described bypeck. mycologia 34: 154 179. z h a n g y. h., z h u a n g w. y. 2004. phylogenetic relationships of some members in the genus hymeno scyphus (ascomycetes, helotiales). nova hedwigia 78: 475 484. hymenoscyphus subcarneus, mało znany miseczniak występujący na mszakach, zebrany w białowieskim parku narodowym s t r e s z c z e n i e hymenoscyphus subcarneus został zebrany jako pasożytujący na wątrobowcu cephalozia catenulata w białowieskim parku narodowym; został opisany i zilustrowany na podstawie świeżych okazów. dwie, nieznane dotychczas cechy zostały opisane: worki z pierścieniem api kalnym typu calycina oraz charakterystyczna zawartość żywych parafiz. ponadto studiowano również zielnikowe okazy pohlia gracilis z grimsel w szwajcarii, na których odkryto nowy rodzaj roseodiscus. autorzy dyskutują wzajemne relacje i taksonomię tego grzyba z podobnymi gatunkami występującymi na pędach skrzypów hymenoscyphus rhodoleucus i h. equisetinus. rozpatry wane są też podobieństwa i różnice między roseodiscus a bliskimi rodzajami jak calycina, stamnaria, rhizoscyphus i pezoloma. 2014-01-01t11:43:08+0100 polish botanical society macrofungi of wooded patches in the agricultural landscape. i. species diversity anna kujawa institute for agricultural and forest environment, polish academy of sciences field station in turew, szkolna 4, pl-64-000 kościan, ankujawa@man.poznan.pl kujawa a.: macrofungi of wooded patches in the agricultural landscape. i. species diversity. acta mycol. 44 (1): 49–75, 2009. this article begins a four-part series, which presents the results of mycological research carried out in 2000-2007 in the agricultural landscape of the general dezydery chłapowski landscape park. this part includes description of the study area and field research methods, as well as a list and localities of 617 macrofungal taxa recorded in the park. the next parts of this series will deal with: species that are rare, protected by law or recorded for the first time in poland; the role of wooded patches for preservation of fungal diversity in the agricultural landscape; and changes in species diversity and structure of fungal communities in forest communities under strong human pressure. key words: macrofungi, nature conservation, agricultural landscape, species diversity introduction a review of polish mycological literature shows a relatively small number of studies dealing with macrofungi occurring in non-farmed habitats in farmlands, which are under strong human pressure. this group of habitats includes small wooded patches (shelterbelts or clumps of trees surrounded by farmland), avenues (roadside verges with trees), village parks, and managed forests (mainly monocultures). they are forest islands surrounded by crop fields, which often play a key role for biodiversity protection in farmland (e.g., ryszkowski at al. 1999; banaszak 2002). mycological studies of small isolated wooded patches by strongly influenced of human activity have been very rarely carried out in poland. only few mycological papers deal with elements of agricultural landscape, e.g., village parks (lisiewska, ratyńska 1984; lisiewska, rybak 1990; lisiewska, płaczek 1993; bujakiewicz, kujawa 2000), orchards (lisiewska, balcerkiewicz 1991), edges of small water bodies in farmland (adamczyk 1997), pastures (domański z. 1969), and forests developing in old fields acta mycologica vol. 44 (1): 49–75 2009 50 a. kujawa (kałucka 1999). there are no complex studies of wooded patches typical for the agricultural landscape, i.e., varying in age, stage of development, origin, etc. such studies seem necessary in the context of reports on disappearance of fungi in europe and the need for protection of this group of organisms (e.g., koune 2001; senn-irlet et al. 2007). this applies to the polish mycobiota, too (e.g., ławrynowicz 1991; grzywacz 1989; łuszczyński 2002). the objective of this study was to describe the species diversity of macrofungi in small wooded patches, village parks and managed forests in the general dezydery chłapowski landscape park. this article begins a four-part series, which presents the results of mycological research conducted in the park. study area the general dezydery chłapowski landscape park (17 200 ha) is located in central wielkopolska, 50 km south of poznań (fig. 1). it was founded in 1992 to protect the unique agricultural landscape markedly enriched with a system of various wooded patches, planted in the early 19th century by dezydery chłapowski, then the owner of the residence in turew. they are protected as a cultural assets at present (ryszkowski 1998, rozporządzenie... 1992). the park consists mainly of crop fields (65.5%), woodlands (15%) and grasslands (8.5%) (rozporządzenie... 1992, fig. 1). it lies in subprovince pojezierze południowobałtyckie lakeland (315), macroregion pojezierze leszczyńskie lakeland (315.8), within 2 mesoregions – pojezierze krzywińskie lakeland (315.82) and równina kościańska plain (315.83) (kondracki 2002). the northern part of the park lies in równina kościańska plain and the southern part belongs to pojezierze krzywińskie lakeland, in which there are several tens of lakes created during the last glacial period, i.e., vistulian glaciation (kondracki 2002). the most common kinds of soils are luvisols (lessive soils) which are a type of autogenic brown-earth soils. they are used mostly for cultivation of various crops. only the valleys of watercourses (kościan obra canal, wyskoć ditch, and smaller watercourses) are filled with alluvial sediments covered with peat. the upper layers of peat have been partly decomposed and the hydrogenic soils formed there are histosol, classified as post-bog soils and muck soil type; peat-muck soils dominate among them (margowski et al. 1976; marcinek 1996). the park lies in the catchment of the obra river, which is an area threatened by water deficit (pasławski 1990). no watercourse in the area has a natural route, as all watercourses were regulated, mainly in the 19th century (kasprzak, raszka 2007). there are relatively numerous small astatic water bodies. for 40 years, disappearance of small water bodies and watercourses has been observed as a result of natural succession, drying out, and human activity (ryszkowski 1990). the growing season lasts 225 days, from late march till the end of october (woś, tamulewicz 1996). mean yearly precipitation (ca. 600 mm) is one of the lowest not only in poland but also in europe. maximum monthly precipitation is observed in july (80 mm), while minimum in february (30 mm) (kędziora, palusiński 1998). macrofungi of wooded patches 51 the area of the park is potentially predominated by galio sylvatici-carpinetum (wojterski et al. 1981; bałazy et al. 1990) (fig. 1) where anthropogenic communities are most common: first of all crop fields, meadows, and managed forests (ratyńska-nowak 1986; ratyńska 1990, bałazy et al. 1990). besides galio sylvaticicarpinetum, also 5 other potential habitats were recorded: salici-populetum, carici elongatae-alnetum, fraxino-alnetum, querco-ulmetum minoris, and calamagrostio arundinaceae-quercetum petraeae (bałazy et al. 1990; ratyńska 1990). forests and smaller wooded patches cover only ca. 15% of the park (2198 ha) and comprise ca. 50 isolated complexes. such forest fragmentation causes higher susceptibility of those complexes to human pressure (olaczek 1972; bałazy et al. 1990). in spite of the potential dominance of the galio sylvatici-carpinetum habitats, managed forest stands very rarely resemble natural forest communities in species composition and vertical structure. there are many degraded forest communities in the park (ratyńska 1990). among the species that are native to wielkopolska, some species that are alien to this region have been planted e.g., picea abies, larix sp., pseudotsuga taxifolia, robinia pseudoacacia, and quercus rubra. moreover, aesculus hippocastanum, acer negundo, and gleditsia triacanthos are often found in avenues and small wooded patches surrounded by crop fields. a characteristic and distinctive feature of the landscape in the park is the presence of trees along field edges, which constitute a system of ecological corridors. since the 1990s, the system has been complemented and reconstructed (kujawa 1998). in the mostly deforested landscape of the park, these wooded patches substitute for forests and compensate for their lack or complement them in biocoenotic functions (bałazy et al. 1990). the first report on macrofungi was published by goszczyński et al. (1980) and dealt with a single record of langermannia gigantea in the palace park in turew. a study of species composition of macrofungi in the village park in turew was carried out in 1997-1999 (bujakiewicz, kujawa 2000), and macrofungal species diversity in 3 belts of various age was evaluated in 1998-2000 (strakulska 2001; lisiewska, strakulska 2002). since 1998 the rate of colonisation by macrofungi of 4 young shelterbelts planted in former crop fields has been studied (kujawa 2007; kujawa, kujawa 2008). there are very few reports on selected species (kujawa, karg 1997; danielewicz, maliński 1999; kujawa 2003a, b; kujawa et al. 2004; ronikier 2005, 2005a; kujawa, karasiński 2007). a preliminary description of mycobiota of the park has been presented by kujawa (2008a). material and methods mycological research was carried out in 2000-2007 (kujawa 2008, kujawa 2008a). the study dealt with ascomycota and basidiomycota according to the classification by hibbett et al. (2007). controversial genera, regarded by some mycologists as microfungi, were included into analyses following hansen and knudsen (2000). data were collected in 50 permanent plots in 2000-2002 and with the route method (searching while walking along a route) in 2000-2007. the plots (400 m2 each) were macrofungi of wooded patches 53 of those communities for preservation of fungal species diversity in farmland. the collected data, complemented with published materials, reflect differentiation of mycobiota in the park and the current status of knowledge. results as many as 615 taxa of macrofungi representing two phyla were recorded in the park. most species (528; 86%) belonged to the phylum basidiomycota, represented by 3 classes, 14 orders, and 52 families. species from the order agaricales constituted 63.5% of all recorded species. the phylum ascomycota (87 species, 14%) was represented by 5 classes, 7 orders and 18 families. richest in recorded species were the orders pezizales (35 spp.) and helotiales (34 spp.). the area of the park is covered by 223 squares (1 km2 each) of the atpol grid; among them, 58 squares lie on the border of the park. the data on fungi were gathered in 75 squares (33.6%). some of them were visited regularly (study in permanent plots), whereas others only sporadically (47 squares). the number of recorded species per square ranged between 1 and 213 (fig. 3). merely 8 squares (3.6%), i.e. those with species number higher than 100, can be regarded as mycologically relatively well-studied. many species (399; 64%) were recorded in only 1-3 squares. according to the criteria adopted in this paper (classes of frequency values), these species are regarded as rare or very rare. among them, 213 species are known from single squares. in highest numbers of squares, the following species were observed: gymnopus dryophilus (28), mycena sanguinolenta (25), xerocomus chrysenteron (24), mycena galericulata (23), gymnopus peronatus (21), marasmius oreades (20), and mycena leptocephala (20). list of species the list below enumaerates all the taxa of macrofungi recorded in the park in 2000– 2007, including the species known from publications dealing with the area of the park. among them, 416 were found on pernament plots and 466 outside in permanent plots (including 153 not recorded in permanent plots). twenty species were found only during the earlier study in the village park in turew (bujakiewicz, kujawa 2000), and 26 are known only from the earlier studies carried out in 3 shelterbelts (strakulska 2001; lisiewska, strakulska 2002). the nomenclature for ascomycota follows chmiel (2006), hansen and knudsen (2000), and index fungorum (2004), while for basidomycota knudsen and vesterholt (2008) and legon et al. (2005) or the monographs of some genera. distribution of species is described with the use of atpol grid (zając 1978), adapted for fungi by wojewoda (2000). species were grouped into functional groups according to the 54 a. kujawa classification by lisiewska (2000), with a slight modification, abbreviations according to friedrich (2001): ectomycorrhizal symbionts (m/s);• humicolous saprotrophs (s/s), including a few bryophilous species (b); • litter-inhabiting saprotrophs, growing on the upper and loose layer of litter, con-• sisting of fallen leaves, seeds, fruits, other dead parts of herbs, mosses, fungi, animals, as well as tiny twigs [although lisiewska (2000) regarded species growing on tiny twigs as lignicolous saprotrophs] (s/l); lignicolous saprotrophs, developing on branches, pieces of bark, stumps and logs • (s/w); parasites, developing on live trees (p/w), herbs (p/p), insects (p/i) or fungi (p/f).• ectomycorrhizal symbionts were distinguished according to the literature (agerer 1987-2008; rudawska 1990, 1993, 2006; wojewoda 2003; mleczko 2004; tedersoo et al. 2006). for each species, the following data are given: functional group, year of observation, reference, and location (tabs 1 and 2). table 1 major characteristics of permanent plots in the general dezydery chłapowski landscape park no. atpol db forest or locality name forest division category of community 1 38-85 błociszewo park i 2 38-85 błociszewo park ii 3 38-86 błociszewo av. aesc 4 47-37 choryń park i 5 47-37 choryń park ii 6 48-13 ćwikłowo 154a mon. frax 7 48-03 ćwikłowo 154c mon. pop 8 37-79 gołębin st. clump 9 37-87 gołębin st. av. q-ro 10 37-69 gorzyczki clump 11 47-45 racot 181c mon. carp 12 48-60 kopaszewo clump 13 48-60 kopaszewo clump 14 48-60 kopaszewo park i 15 48-60 kopaszewo park ii 16 48-70 kopaszewo av. aesc 17 47-59 kopaszewo 165c mon. fag 18 47-02 racot park i 19 47-02 racot park ii 20 47-03 racot av. til 21 47-44 racot 191j mon. bet 22 48-04 rąbiń 128h mon. rob 23 48-04 rąbiń 130c mon. aln 24 48-23 rąbiń av. til 25 48-05 rąbiń 126c mon. lar 26 48-17 rąbiń 136j mon. pic 27 48-34 rąbiń shelterbelt 28 48-17 rąbiń 142a mon. pse 29 48-05 rąbiń 127b mon. aln 30 48-05 rąbiń 127g mon. bet 31 48-05 rąbiń 127d mon. frax 32 48-30 turew shelterbelt 33 48-11 turew av. q-ro 34 48-00 turew shelterbelt macrofungi of wooded patches 55 no. atpol db forest or locality name forest division category of community 35 48-00 turew park i 36 48-10 turew park ii 37 38-91 turew shelterbelt 38 38-90 turew 133i mon. q-ro 39 38-90 turew 133b mon. pop 40 38-91 turew 133d mon. pin 41 38-91 turew 133g mon. lar 42 38-90 turew 134h mon. rob 43 38-90 turew 134d mon. carp 44 38-90 turew 135j mon. q-ru 45 38-90 turew 134c mon. q-ru 46 38-90 turew 134g mon. pic 47 38-91 turew 133d mon. til 48 38-81 wronowo 132a mon. fag 49 47-09 wyskoć 159a mon. pin 50 47-08 wyskoć 160b mon. q-ro abbreviations: av. = avenue; mon. = forest monoculture; park i = plot in cultivated part of a village park; park ii = plot in uncultivated part of a village park. species of trees: aesc = aesculus hippocastanum; aln = alnus glutinosa; bet = betula pendula; carp = carpinus betulus; fag = fagus sylvatica; frax = fraxinus excelsior; lar = larix decidua; pic = picea abies; pin = pinus sylvestris; pop = populus sp.; pse = pseudotsuga taxifolia; q-ro = quercus robur; q-ru = quercus rubra; rob = robinia pseudacacia; til = tilia cordata table 2 localities (atpol squares) of records outside the permanent plots no. atpol db no. atpol db no. atpol db no. atpol db no. atpol db 51 37-44 66 38-90 81 47-39 96 48-04 111 48-24 52 37-45 67 38-91 82 47-44 97 48-05 112 48-25 53 37-69 68 38-92 83 47-45 98 48-07 113 48-26 54 37-76 69 38-93 84 47-46 99 48-08 114 48-27 55 37-77 70 47-02 85 47-48 100 48-10 115 48-30 56 37-79 71 47-03 86 47-54 101 48-11 116 48-32 57 37-84 72 47-05 87 47-55 102 48-12 117 48-34 58 37-88 73 47-08 88 47-56 103 48-13 118 48-37 59 37-95 74 47-09 89 47-58 104 48-14 119 48-51 60 37-98 75 47-17 90 47-59 105 48-15 120 48-60 61 37-99 76 47-19 91 47-79 106 48-16 121 48-61 62 38-68 77 47-28 92 48-00 107 48-17 122 48-70 63 38-81 78 47-33 93 48-01 108 48-21 123 48-71 64 38-85 79 47-34 94 48-02 109 48-22 124 48-92 65 38-86 80 47-37 95 48-03 110 48-23 125 49-01 in species list, the following abbreviations were used: threat categories: ex = extinct or probably extinct, e = endangered, v = vulnerable, r = rare, i = indeterminate, p = protected by law (wojewoda, ławrynowicz 2006), * new species in poland functional groups: m/s = mycorrhizal fungi, s/s = humicolous saprotrophs, s/l = litter-inhabiting saprotrophs, s/w = lignicolous saprotrophs, p/p = parasites of herbaceous plants, p/f = parasites of fungi, p/i = parasites of insects, p/w = parasites of woody plants, (b) = bryophilous species. published data sources: b&k 2000 = bujakiewicz and kujawa 2000; d&m 1999 = danielewicz and maliński 1999; g et al. 1980 = goszczyński et al. 1980; k&k 1997 = karg and kujawa 1997; k 2003a = kujawa 2003a; k 2003b = kujawa 2003b; k et al. 2004 = kujawa et al. 2004; l&s 2002 = lisiewska and strakulska 2002; r 2005a = ronikier 2005a; r 2005b = ronikier 2005b; s 2001 = strakulska 2001 56 a. kujawa ascomycota aleuria aurantia (pers.) fuckel – s/s; 1997, 1999: (b&k 2000), 2000, 2007: 99, 107. ascocoryne cylichnium (tul.) korf – s/w; 1998: (b&k 2000); a. sarcoides (jacq.) j.w. groves & d.e. wilson – s/w; 2001-2002, 2007: 11, 21, 92, 103. bisporella citrina (batsch) korf & s.e. carp. – s/w; 2000-2002: 38, 45. bulgaria inquinans (pers.) fr. – s/w; 1999: (b&k 2000). ciboria amentacea (balb.) fuckel – s/l; 2006: 100; c. batschiana (zopf) n.f. buchw. – s/l; 1999: (b&k 2000), 2000-2001: 33, 36, 38, 50; c. viridifusca (fuckel) höhn. – s/l; 1997: (b&k 2000). cistella acuum (alb. & schwein.) svrček – s/l; 2000: 40. cordyceps capitata (holmsk.) fr. – p/f, r; 2000-2001: 11, 45; c. entomorrhiza (dicks.) fr. – p/i; 2002: 6; c. ophioglossoides (ehrh.) fr. – p/f, r; 2000: 50. crocicreas coronatum (bull.) s.e. carp. – s/l; 1999: (b&k 2000), 2000-2001, 2003, 2005: 6, 8 10, 12, 23, 58; c. cyathoideum (bull.) s.e. carp. var. cyathoideum – s/l; 2000: 27. daldinia concentrica (bolton) ces. & de not. – s/w; 2002: 39. *desmazierella cf. piceicola huhtinen & y. mäkinen – s/l; 2000: 26. discina ancilis (pers.) sacc. – s/w, r; 1998: 74. dumontinia tuberosa (hedw.) l.m. kohn – p/p; 1999 (b&k 2000); 2000, 2002, 2004, 2005: 14, 15, 92, 100. elaphomyces granulatus em. hollós – m/s; 2000: 50; e. muricatus fr. – m/s; 2000, 2001: 11, 45. encoelia furfuracea (roth) p. karst. – s/w; 2007: 100. *geopora foliacea (schaeff.) s. ahmad – s/s; 2003-2004, 2007: 74. gyromitra esculenta (pers.) fr. – s/s; 2001: 106. helvella acetabulum (l.) quél. – s/s; 2002: 60; h. atra j. köenig – s/s; 2004: 107; h. crispa (scop.) fr. – s/s; 1997: (b&k 2000), 2000, 2003: 100; h. elastica bull. – s/s; 1998: (b&k 2000), 2006: 100; h. ephippium lév. – s/s, r; 2000-2001: 8, 34; h. lacunosa afzel. – s/s, r; 2000-2001, 2006: 48, 50, 58, 107; h. macropus (pers.) p. karst. – s/s; 1998: (b&k 2000), 2000-2001: 21, 30, 34, 39, 47. humaria hemisphaerica (f.h. wigg.) fuckel – s/s; 1998: (b&k 2000), 2000-2001, 2003, 2005: 9, 11, 13, 29, 30, 34, 47, 48, 50, 100, 107. hymenoscyphus albidus (roberge ex desm.) w. phillips – s/l; 2001-2002: 13, 15, 29, 31, 58, 97; h. calyculus (sowerby) w. phillips – s/w; 2000: 97; h. epiphyllus (pers.) rehm ex kauffman – s/l; 2000: 49; h. fructigenus (bull.) fr. – s/l; 1997-1999: (b&k 2000), 2000-2004: 1, 2, 4, 5, 9, 11, 15, 16, 18, 27, 33, 36, 50, 73, 100, 120, 125; h. herbarum (pers.) dennis – s/l; 2000: 29; h. imberbis (bull.) dennis – s/w; 2000: 23; h. immutabilis (fuckel) dennis – s/l; 2000-2001: 22, 40, 42; h. phyllophilus (desm.) kuntze – s/l; 2000: 65; h. scutula (pers.) w. phillips – s/l; 1999: (b&k 2000), 2000-2007: 6, 7, 8, 9, 10, 27, 31, 35, 58, 60, 73. hypoxylon fragiforme (pers.) j. kickx f. – s/w; 1998: (b&k 2000), 2001, 2003, 2007: 43, 83, 92; h. howeanum peck – s/w; 2000-2002: 11, 31, 43; h. multiforme (fr.) fr. – s/w; 2001: 23. lachnellula willkommii (hartig) dennis – s/w; 2000, 2002: 25. macrofungi of wooded patches 57 lachnum brevipilosum baral – s/w; 2002: 31; l. virgineum (batsch) p. karst. – s/l; 2000-2004: 12, 14, 16, 17, 36, 48, 60. lanzia luteovirescens (roberge ex desm.) dumont & korf – s/l; 1999: (b&k 2000), 4, 5, 18, 19, 97. melastiza chateri (w.g. sm.) boud. – s/s; 2002: 56. mollisia amenticola (sacc.) rehm – s/l; 1997: (b&k 2000), 2000: 10; m. cinerea (batsch) p. karst. – s/w; 1999: (b&k 2000), 2000-2001: 14, 24, 27, 29, 30. *monilinia johnsonii (ellis & everh.) honey – s/l; 2000: 27. morchella conica pers. – m?/s, p, r; 2000-2003 (k 2003b), 2004-2005: 93, 106; m. esculenta (l.) pers. – m?/s, p, r; 2005: 100; m. gigas (batsch) pers. [= mitrophora semilibera (dc.) lév.] – m?/s, p, r; 1998-2002: (k&k 1997, b&k 2000, k 2003b), 2000, 2002-2005: 6, 31, 34, 60, 74, 121. nectria cinnabarina (tode) fr. – s/w; 2001-2003, 2007: 1, 2, 3, 4, 5, 12, 14, 16, 17, 18, 21, 23, 30, 38, 46, 47, 58, 69, 73. octospora humosa (fr.) dennis – s/s, (b); 2004-2005: 105, 107. orbilia xanthostigma (fr.) fr. – s/w; 1997: (b&k 2000). otidea alutacea (pers.) massee – s/s; 1997, 1999: (b&k 2000), 2000, 2003: 100; o. onotica (pers.) fuckel – s/s; 2000: 50. *peziza ampliata pers. – s/w; 2000-2002: 47; p. arvernensis boud. – s/s; 2002: 17; p. badia pers. – s/s; 2000: 100; p. depressa pers. – s/s; 2000-2002: 15, 34, 100; p. howsei (boud.) donadini – s/s; 200-2001, 2005-2006: 15, 34, 114; p. micropus pers. – s/w; 1997-1998: (b&k 2000), 2000-2002, 2006: 92, 100, 117; p. repanda pers. – s/s; 2000: 39; p. succosa berk. – s/s; 2000-2001, 2003: 15, 18, 34, 92; p. vesiculosa bull. – s/s; 1999-2000: (s 2001, l&s 202), 2000, 2004, 2007: 73, 92. pezizella alniella (nyl.) dennis – s/l; 2006: 100. *pustularia patavina (cooke & sacc.) boud. – s/s; 2005-2006: 60. rhytisma acerinum (pers.) fr. – p/s/l; 2001-2007: 58, 73. rutstroemia bulgarioides (rabenh.) p. karst. – s/l; 2001: 26, 46; r. conformata (p. karst.) nannf. – s/l; 1999: (b&k 2000), 2000: 29; r. sydowiana (rehm) w.l. white – s/l; 1999: (b&k 2000), 2000-2002: 14, 27, 38, 45, 50, 107. sclerotinia trifoliorum erikss. – p/p; 2005: 60. scutellinia scutellata (l.) lambotte – s/w;1997-1998: (b&k 2000), 2001-2004: 13, 29, 35, 92, 93, 96, 97, 120; s. trechispora (berk. & broome) lambotte – s/s; 2000: 15. tapesia fusca (pers.) fuckel – s/w; 2000-2001: 13, 15, 17, 29, 43, 44, 47, 80. tarzetta cupularis (l.) lambotte – s/s; 1998: (b&k 2000), 2000-2002, 2004: 8, 15, 18, 19, 23, 24, 29, 31, 35, 48, 100, 107. trichophaea gregaria (rehm) boud. – s/s; 2006-2007: 58, 60. ustulina deusta (hoffm.) lind [=kretzschmaria deusta (hoffm.) p.m.d. martin] – s/w; 2002-2003, 2007: 17, 19, 20, 45, 48, 92. verpa conica (o.f. müll.) sw. – s/s, p, r; 1999-2000, 2002: (k 2003b), 2004: 100. xylaria carpophila (pers.) fr. – s/l; 2000-2004: 17, 35, 36, 48; x. hypoxylon (l.) grev. – s/w; 1997-1998: (b&k 2000), 2000-2003, 2005, 2007: 2, 4, 6, 11, 12, 15, 17, 18, 19, 26, 31, 36, 48, 100; x. longipes nitschke – s/w; 2004: 63; x. oxyacanthae tul. & c. tul. – s/l; 2001-2006: 34, 92; x. polymorpha (pers.) grev. – s/w; 1999: (b&k 2000), 2000-2002: 11, 15, 17, 19, 36, 47. 58 a. kujawa basidiomycota agaricus arvensis schaeff. – s/s; 1999-2000: (b&k 2000, s 2001, l&s 2002), 20002007: 3, 7, 12, 20, 24, 33, 58, 65, 73, 80, 100; a. augustus fr. – s/s; 2002, 2004: 100, 120; a. bitorquis (quél.) sacc. – s/s; 2000, 2001-2003, 2006: 20, 24, 33, 80, 100, 125; a. campestris l. – s/s; 1998 (s 2001, l&s 2002), 2001-2002, 2004: 6, 73; a. comtulus fr. – s/s, r; 2000, 2003-2004, 2007: 73, 92; a. essettei bon – s/s; 2004: 92; *a. litoralis (wakef. & a. pearson) pilát – (leg. k. kujawa) s/s; 2002: 123; a.semotus fr. – s/s; 1998: (b&k 2000); a. sylvaticus schaeff. – s/s; 2000-2001: 12, 13, 97; a. sylvicola (vittad.) peck – s/s; 1998-2000 (s 2001, l&s 2002), 2000-2002, 2005-2007: 2, 6, 32, 58, 73; a. xanthodermus genev. – s/s; 1998, 2000: (s 2001, l&s 2002), 2000, 2002-2004, 2006-2007: 35, 60, 70, 74, 80, 100, 120. agrocybe arvalis (fr.) r. heim & romagn. – s/s; 2000 (s 2001, l&s 2002), 2000: 73; a. cylindrica (dc.) maire – s/s, e; 1999: (b&k 2000); a. dura (bolton) singer – s/s; 2000-2002, 2006-2007: 58, 60; a. pediades (fr.) fayod – s/s; 2001-2007: 27, 58, 60; a. praecox (pers.) fayod – s/s; 2000: (s 2001, l&s 2002), 2000, 2002, 20042005: 7, 22, 58, 60, 73; a. sphaleromorpha (bull.) fayod (according to legon et al. 2005 it is one of the species belonging to a. praecox complex) – s/s; 2000: (s 2001, l&s 2002). amanita citrina (schaeff.) pers. f. citrina – m/s; 1997: (b&k 2000), 2000-2002: 11, 17, 43, 44, 45, 50, 90; a. excelsa (fr.) p. kumm. f. excelsa – m/s; 2000-2001: 43, 106; a. fulva (schaeff.) fr. – m/s; 2000-2002: 28, 45, 50, 66, 114; a. muscaria (l.) lam. var. muscaria – m/s; 2000: (s 2001, l&s 2002), 2000-2007: 17, 21, 28, 73, 90, 106; a. pantherina (dc.) krombh. – m/s; 2000, 2003, 2006: 9, 43, 80, 100; a. phalloides (vaill.) link var. phalloides – m/s; 2000-2001: 9, 11, 24, 33, 43, 63, 90, 100; a. porphyria alb. & schwein. – m/s; 2001, 2003: 26, 113; a. rubescens pers. f. rubescens – m/s; 1997 (b&k 2000), 2000-2004: 2, 5, 11, 17, 28, 35, 43, 44, 45, 50, 66, 90, 100, 109; a. strobiliformis (vittad.) bertill. – m/s, 2007: 100; a. vaginata (bull.) lam. f. vaginata – m/s; 2001: 66. ampulloclitocybe [clitocybe] clavipes (pers.) redhead, lutzoni, moncalvo & vilgalys (rev. h. komorowska) – s/s; 1997, 2000: (b&k 2000, s 2001, l&s 2002), 20002003: 16, 25, 26, 28, 38, 40, 41, 49, 82, 86, 114. armillaria borealis marxm. & korhonen – p/w; 2000-2001: 11; a. cepistipes velen. – p/w; 2004: 100; a. lutea gillet – p/w; 1997-1998 (b&k 2000, as: a. mellea s.l.), 2000-2001: 1, 4, 5, 15, 18, 28, 30, 35, 36, 38, 42, 45, 46; a. mellea (vahl.) p. kumm. s.str. – p/w; 2000, 2004-2006: 92; a. ostoyae (romagn.) herink – p/w; 2003-2006: 66, 100, 112. auricularia auricula-judae (bull.) wettst. – s/w; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2003: 1, 2, 4, 5, 8, 10, 12, 14, 19, 27, 36, 37, 69. auriscalpium vulgare gray – s/l; 2000-2003, 2005-2007: 23, 26, 28, 40, 49, 50, 58, 68, 73, 82, 83, 86, 114. baeospora myosura (fr.) singer – s/l; 2001-2004: 26, 40, 46, 47, 49, 79. bjerkandera adusta (willd.) p. karst. – s/w; 1999-2000: (b&k 2000, s 2001, l&s 2002), 2000-2001, 2003, 2007: 21, 26, 62, 83, 100. bolbitius titubans (bull.) fr. – s/l; 1999-2000: (s 2001, l&s 2002), 2000-2003: 3, 8, 24, 39, 52, 73, 82, 100, 103, 114. macrofungi of wooded patches 59 boletus edulis bull. – m/s; 2000-2004, 2006: 9, 21, 24, 43, 44, 48, 50, 66, 73, 83, 90, 100, 114; b. luridiformis rostk. var. luridiformis – m/s; 2000, 2006: 73, 106; b. luridus schaeff. var. luridius – m/s; 2000-2001, 2004: 9, 24, 48, 63. bovista dermoxantha (vittad.) de toni – s/s; 1998-1999: (s 2001, l&s 2002), 20002004, 2007: 2, 4, 5, 16, 20, 27, 30, 32, 47, 48, 60, 73, 100, 114; b. plumbea pers. – s/s; 1999-2000: (s 2001, l&s 2002); 2002, 47; b. nigrescens pers. – s/s; 2000, 2003: 6, 7, 30, 67, 68, 118. byssomerulius corium (pers.) parmasto – s/w; 1997: (b&k 2000), 2007: 58. calocera cornea (batsch) fr. – s/w; 2000-2003: 6, 11, 17, 39, 43, 44, 45; c. viscosa (pers.) fr. – s/w; 2000-2003: 25, 26, 28, 41, 46, 62, 66, 82, 80, 86, 98, 100, 114. calocybe gambosa (fr.) donk f. gambosa – s/s; 2000-2006: 3, 6, 8, 20, 27, 60, 114, 120. calvatia [langermannia] gigantea (batsch) lloyd – s/s, p; (d&m 1999, k&k 1997, k 2003b), 1970: (g et al. 1980, k&k 1997), 1997, 2000, 2002: (b&k 2000, k 2003b), 2005-2006: 19, 77 (leg., det. h. gołdyn), 81 (leg., det. d. sobczyk), 84, 91, 100, 120 (leg., det. e. górna). calyptella capula (holmsk.) quél. – s/l, r; 2000-2001: 31, 35. cantharellus cibarius fr. – m/s; 1987-1996: (k&k 1997), 2000-2003: 28, 40, 44, 45, 50, 62, 66, 73, 114. chalciporus piperatus (bull.) bataille – m/s; 2000-2001: 28, 45, 90. chlorophyllum brunneum (farl. & burt.) vellinga [= macrolepiota rhacodes var. bohemica (wichanský) bellù & lanzoni] – s/s; 1998, 2000: (s 2001, l&s 2002); ch. [macrolepiota] rhacodes (vittad.) vellinga – s/s; 1998-1999: (b&k 2000, s 2001, l&s 2002), 2000-2002, 2006: 1, 2, 3, 4, 12, 16, 20, 26, 27, 32, 36, 37, 38, 41, 46, 60, 74, 90, 100, 103. chondrostereum purpureum (pers.) pouzar – s/w; 1999: (b&k 2000), 2001-2002: 21. *clavicorona taxophila (thom) doty – s/s; 2000-2001: 7. clavulina cinerea (bull.) j. schröt. – s/s; 1997-1998: (b&k 2000), 2000-2001, 2003: 48, 100; c. coralloides (l.) j. schröt. – s/s; 2001, 2003: 7, 114. clitocybe agrestis harmaja – s/l, r; 1999: (s 2001, l&s 2002); c. anisata velen. (det. h. komorowska) – s/l; 2001: 115; c. candicans (pers.) p. kumm. var. candicans – s/l; 1999: (s 2001, l&s 2002), 2000, 2007: 2, 58, 73; c. cerussata (fr.) p. kumm. (according to legon et al. 2005 it is synonym of c. phyllophila) – s/l; 2000: (s 2001, l&s 2002); c. ditopus (fr.) gillet (rev. h. komorowska) – s/l; 2000-2002: 27, 36; c. fragrans (with.) p. kumm. – s/l; 1998-2000: (s 2001, l&s 2002, as: c. fragrans et c. suaveolens); c. metachroa (fr.) p. kumm. var. metachroa – s/l; 1998, 2000: (s 2001, l&s 2002), 2001, 2004, 2007: 73; c. nebularis (batsch) p. kumm. var. nebularis – s/s; 1998: (s 2001, l&s 2002), 2000-2002, 2004, 2007: 2, 4, 5, 19, 20, 21, 36, 37, 38, 39, 47, 58, 73, 78, 80, 115; c. odora (bull.) p. kumm. var. odora – s/l; 2000: (s 2001, l&s 2002), 2000, 2002-2003, 2005-2007: 2, 21, 23, 38, 73, 100; c. phaeophthalma (pers.) kuyper (rev. h. komorowska) – s/l, r; 2000-2002: 32, 37, 38, 42; c. phyllophila (pers.) p. kumm. (rev. h. komorowska) – s/l; 2001: 4, 21; c. rivulosa (pers.) p. kumm. [= c. dealbata (sowerby) p. kumm.] – s/l; 19982000: (s 2001, l&s 2002), 2000, 2007: 20, 58; c. trulliformis (fr.) p. karst. (det. h. komorowska) – s/s; 2000: 95; c. vermicularis (fr.) quél. – s/l; 1998: (s 2001, l&s 2002); c. vibecina (fr.) quél. – s/l; 2000: (s 2001, l&s 2002), 2001, 2004: 22, 26, 73. 60 a. kujawa clitopilus prunulus (scop.) p. kumm. – m/s; 2000-2002: 20, 21, 48. collybia cirrata (pers.) quél. – s/l; 2001, 2003-2005, 2007: 46, 60, 73; c. cookei (bres.) j.d. arnold – s/l; 2001, 2003-2005, 2007: 13, 73, 92; c. tuberosa (bull.) p. kumm. – s/l; 2001: 50. conocybe albipes (g.h. otth) hauskn. – s/s; 2002, 2005: 60, 120; c. mesospora kühner & watling – s/s; 2000-2001, 2007: 60; c. pilosella (pers.) kühner – s/s; 2000: 31; c. pubescens (gillet) kühner – s/s; 2000: 60; c. rickeniana p.d. orton – s/s; 1997: (b&k 2000), 2000-2002, 2005-2006: 3, 7, 12, 13, 15, 18, 19, 34, 73; c. semiglobata kühner & watling – s/s; 2000-20004, 2007: 8, 10, 58, 60, 73, 117; c. subovalis kühner & watling – s/s; tu, 2007: 60; c. subpubescens p.d. orton – s/s; 2001-2002: 33, 38; c. tenera (schaeff.) fayod – s/s; 1998-2000: (s 2001, l&s 2002), 2000-2003: 7, 13, 27, 33, 34, 39, 100, 115; c. velutipes (velen.) hauskn. & svrček – s/s; 2000, 2003: 37, 79. coprinellus [coprinus] angulatus (peck) redhead, vilgalys & moncalvo – s/s, 2002: 67; c. disseminatus (pers.) j.e. lange – s/w; 1998-2000: (b&k 2000, s 2001, l&s 2002), 2000-2004, 2006: 6, 8, 10, 12, 19, 30, 31, 34, 80, 92; c. domesticus (bolton) vilgalys, hopple & jacq. johnson – s/w; 1999-2000: (s 2001, l&s 2002), 2000, 2002-2006: 6, 12, 13, 38, 60, 63, 100, 103, 120; c. impatiens (fr.) j.e. lange – s/s; 1997: (b&k 2000), 2000, 2006: 4, 92; c. micaceus (bull.) vilgalys, hopple & jacq. johnson – s/w; 1998-2000: (b&k 2000, s 2001, l&s 2002), 2000-2004, 2006: 1, 5, 6, 7, 12, 13, 15, 18, 22, 30, 34, 37, 39, 40, 47, 63, 66, 73, 83, 89, 97, 98, 100, 120 ; c. truncorum (schaeff.) vilgalys, hopple & jacq. johnson – s/s; 2007: 74; c. xanthothrix (romagn.) vilgalys, hopple & jacq. johnson – s/w; 1998-2000: (s 2001, l&s 2002), 2001, 2003: 24, 37, 67, 68, 82, 86, 100. coprinopsis [coprinus] atramentaria (bull.) redhead, vilgalys & moncalvo – s/s; 1998-1999 (b&k 2000), 2000-2003: 6, 7, 13, 15, 18, 34, 37, 70, 76, 100; c. erythrocephala (lév.) redhead, vilgalys & moncalvo – s/w; 2000-2001: 34; c. friesii (quél.) p. karst. – s/l; 2000, 2002: 29, 110; *c. insignis (peck) redhead, vilgalys & moncalvo – s/s; 2000, 2004, 2006: 92, 100. coprinus comatus (o.f. müll.) pers. – s/s; 1998-2000: (s 2001, l&s 2002), 2000, 2002-2004, 2006: 37, 61, 74, 75, 76, 100, 109; c. cortinatus j.e. lange – s/s; 1999: (b&k 2000). coriolopsis gallica (fr.) ryvarden – s/w, r; 2002: 31. cortinarius alnetorum (velen.) m.m. moser – m/s; 1998: (b&k 2000), 2001: 36; c. cinnamomeus (l.) gray – m/s; 2000-2001: 26, 28, 49; c. helvelloides (fr.) fr. – m/s; 1998: (b&k 2000); c. hemitrichus (pers.) fr. – m/s; 2000-2001: 39; c. torvus (fr.) fr. – m/s; 2000-2001: 43, 83. craterellus cornucopioides (l.) pers. – m/s; 2006: 114. crepidotus lundellii pilát – s/w; 2001, 2007: 19, 34, 36, 58, 60; c. variabilis (pers.) p. kumm. – s/w; 1999: (b&k 2000), 2000-2003: 10, 11, 19, 29, 30, 34, 36, 38, 42, 43, 45, 47, 50, 82, 83. crinipellis scabella (alb. & schwein.) murrill – s/l; 2000-2003, 2005-2007: 9, 20, 24, 27, 33, 37, 58, 60, 73. crucibulum laeve (huds.) kambly – s/w; 1998-1999: (b&k 2000, s 2001, l&s 2002), 1997, 2000-2003: 1, 2, 5, 11, 35, 36, 70, 81, 100, 114. macrofungi of wooded patches 61 cyathus olla (batsch) pers. – s/l; 1998-2000: (b&k 2000, s 2001, l&s 2002), 20002007: 3, 4, 9,16, 24, 27, 33, 35, 60, 62, 73; c. striatus (huds.) willd. – s/w; 20002004, 2006: 2, 4, 11, 13, 19, 34, 35, 73, 80, 83. cystoderma amianthinum (scop.) fayod – s/s (b); 2000-2002: 25, 50. cystolepiota seminuda (lasch) bon – s/s; 1998: (b&k 2000), 2000-2002: 39, 47. dacrymyces stillatus nees. – s/w; 2001-2002, 2004: 4, 17, 18, 23, 28, 34, 37, 41, 42, 43, 44, 45, 46, 73. daedalea quercina (l.) pers. – s/w; 1998: (s 2001, l&s 2002), 2000, 2002-2003: 38, 45, 83, 86, 90, 114, 125. daedaleopsis confragosa (bolton) j. schröt. – s/w; 2000, 2003: 39, 67, 114. delicatula integrella (pers.) pat. – s/w; 2000-2001: 15, 23, 29, 31. echinoderma [lepiota] aspera (pers.) bon– s/s; 1997: (b&k 2000), 2002: 19; e. echinacea (j.e. lange) bon – s/s, v; 2000-2002: 12, 14, 27, 65. entoloma araneosum (quél.) m.m. moser f. fulvostrigosum (berk. & broome) noordel. – s/s; 1998-1999: (b&k 2000), 2000, 2004: 15, 100; e. byssisedum (pers.) donk – s/w, r; 1999: (b&k 2000); *e. cephalotrichum (p.d. orton) noordel. – s/s; 2001-2002: 38; e. clypeatum (l.) p. kumm. f. clypeatum – m?/s; 2001-2003: 6, 8; e. dysthaloides noordel. – s/s; 2000: 8; e. euchroum (pers.) donk – s/w, r; 2000: 23; e. excentricum bres. – s/s, r; 1999: (s 2001, l&s 2002); e. hirtipes (schumach.) m.m. moser – s/s; 2000, 2004: 6, 16, 100; e. incarnatofuscescens (britzelm.) noordel. – s/s; 2001, 2003, 2006: 100, 103; e. lividocyanulum noordel. – s/s; 2001: 6; e. nitidum quél. – m/s; 2000: 83; *e. parasiticum (quél.) krei-kreisel – (leg. jerzy karg), p/f; 1999: 73; e. percandidum noordel. – s/s; 2001: 25; e. pleopodium (bull.) noordel. – s/l, r; 2000-2001: 30; e. politum (pers.) donk – s/s; 2006: 100; e. rhodocalix (lasch) m.m. moser – s/s, r; 2000-2001: 35, 50; e. rhodopolium (fr.) p. kumm. var. rhodopolium – s/s; 1997, 1999: (b&k 2000, as: e. rhodopolium f. nidorosum et e. rhodopolium), 2000-2001, 2004: 1, 2, 4, 5, 6, 12, 17, 18, 30, 35, 39, 47; e. rusticoides (gillet) noordel. – s/s, e; 2002: 27; e. sericeum quél. var. sericeum – s/s; 1999: (s 2001, l&s 2002); e. turbidum (fr.) quél. – s/s, r; 2002: 28; e. undatum (fr.) m.m. moser – s/s; 2001: 27; e. vernum s. lundell – s/s; 2001, 2006: 39, 114. exidia glandulosa (bull.) fr. [= e. truncata fr.] – s/w, r; 1998, 2000: (b&k 2000, s 2001, l&s 2002), 2000-2003, 2005: 18, 38, 41, 47, 50, 70, 86, 100; e. plana (f.h. wigg.) donk. – s/w; 2001-2003, 2005, 2007: 6, 7, 13, 17, 18, 19, 21, 22, 23, 24, 29, 31, 37, 38, 39, 42, 44, 45, 47, 48, 49, 50, 58, 67, 100, 114. fistulina hepatica (schaeff.) fr. – p/w, p, r, 1987-1999: (k&k 1997, b&k 2000), 2000-2002, 2006: 66, 92, 120. flammulaster carpophilus (fr.) earle – s/l, r; 2002: 17. flammulina velutipes (curtis.) p. karst. var. velutipes – s/w; 1998, 2000: (b&k 2000, s 2001, l&s 2002), 2000, 2002: 21, 31, 47. fomes fomentarius (l.) j.j. kickx – s/w and p/w; 1997: (b&k 2000), 2000-2003: 11, 26, 30, 39, 47, 48, 51, 60, 83, 96, 98, 100, 107, 114, 120. fomitopsis pinicola (sw.) p. karst. – s/w; 2005: 84. galerina clavata (velen.) kühner – s/s, (b); 2000-2006: 58, 60, 73; g. hypnorum (schrank) kühner – s/s, (b); 2000-2001: 26, 28, 46, 50; g. marginata (batsch) kühner s.l. – s/w; 1998: (b&k 2000, as: g. unicolor); g. mniophila (lasch) kühner – s/s, (b); 2001-2002: 25; g. subclavata kühner – s/s, (b); 2003: 73; g. triscopa 62 a. kujawa (fr.) kühner – s/w, r; 2001-2002: 25, 26, 28, 41, 46; g. vittiformis (fr.) singer var. vittiformis f. tetraspora a.h. sm & singer – s/s, (b); 2000-2002: 25, 26, 41. *gamundia striatula (kühner) raithelh. (rev. h. komorowska) – s/s, (b); 20032005, 2007: 92. ganoderma applanatum (pers.) pat. – s/w; 1998: (b&k 2000), 2002-2003, 2005: 31, 67, 83, 84; g. lucidum (curtis) p. karst. – s/w, p, r; (k&k 1997); 2003-2004: 58, 73. geastrum berkeleyi massee – (leg. michał wójtowski), m?/s, p, ex; 2007: 114; g. corollinum (batsch) hollós – (leg. artur golis), m?/s, p, e; 2004, 2007: 63; g. coronatum pers. – m?/s, p, v; 2003-2007: 60; g. fornicatum (huds.) hook. – m?/s, p, e; 2001, 2003-2007: 4, 60; g. rufescens pers. – m?/s, p, e; 2006-2007: 114; g. striatum dc. – m?/s, p, e; 2000-2002: (k 2003a), 2003-2007: 60, 63, 75, 115 (leg. d. sobczyk). gloeophyllum odoratum (wulfen) imazeki – s/w; 1997, 2000: (b&k 2000), 2003-2004, 2007: 100. gomphidius glutinosus (schaeff.) fr. – m/s, r; 2007: 60; g. roseus (fr.) p. karst. – m/s, r; 2005: 106. grifola frondosa (dicks.) gray – p/w, p, v; 1996, 2001: (k&k 1997, k 2003b), 20042007: 92. gymnopilus penetrans (fr.) murrill – s/w; 2000-2002: 40; g. spectabilis (weinm.) a.h. sm. – s/w; 2000-2004: 34, 55, 67, 100. gymnopus [setulipes] androsaceus (l.) antonín & noordel. – s/l; 1998: (s 2001, l&s 2002), 2000-2004: 25, 26, 28, 49, 50, 52, 58, 73, 82, 114; g. aquosus (bull.) antonín & noordel. – s/l; 2004, 2007: 66, 73; g. brassicolens (romagn.) antonín & noordel. – s/l; 2000-2003: 7, 9, 12, 14, 16, 21, 22, 42, 114; g. confluens (pers.) antonín, halling & noordel. – s/l; 1998-1999: (b&k 2000, s 2001, l&s 2002), 2001, 2003, 2005-2007: 100; g. dryophilus (bull.) murrill – s/l; 1997-2000: (b&k 200, s 2001, l&s 2002), 2000-2007: 2, 4, 6, 7, 9, 12, 16, 21, 22, 26, 30, 31, 32, 33, 35, 36, 38, 39, 40, 41, 42, 46, 47, 49, 50, 51, 54, 58, 62, 67, 68, 73, 86, 89, 90, 96, 98, 100, 114, 120; g. erythropus (pers.) antonín, halling & noordel. – s/w; 1999: (s 2001, l&s 2002), 2000-2002: 4, 5, 11, 14, 18, 22, 28, 45; g. fusipes (bull.) gray – p/w; 2006: 114; g. hariolorum (bull.) antonín, halling & noordel. – s/l; 2001: 7; g. hybridus (kühner & romagn.) antonín & noordel. – s/l; 1998: (s 2001, l&s 2002); g. ocior (pers.) antonín & noordel. – s/s, e; 1999: (b&k 2000), 20002003, 2005-2006: 6, 7, 16, 25, 39, 40, 49, 58, 66, 73, 80, 90; g. peronatus (bolton) antonín, halling & noordel. – s/l; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2003: 2, 3, 4, 5, 11, 17, 27, 28, 33, 35, 36, 41, 44, 45, 46, 48, 50, 62, 67, 68, 73, 82, 87, 100, 114. gyrodon lividus (bull.) p. karst. – m/s, r; 2000, 2003: 23, 96. gyroporus castaneus (bull.) quél. – m/s, r; 2000-2001: 83; g. cyanescens (bull.) quél. – m/s, r; 1998, 2001, 2006: 73, 74. handkea excipuliformis (scop.) kreisel – s/s; 1998-2000: (s 2001, l&s 2002), 20002001, 2005, 2007: 5, 7, 12, 24, 32, 37, 39, 58, 73, 88; h. utriformis (bull.) pers. – s/s; 1999: (s 2001, l&s 2002), 2001, 2004, 2007: 58, 111, 112. hapalopilus nidulans (fr.) p. karst. – s/w; 2000-2002: 2, 14, 21, 25, 30, 39, 44, 45, 47, 66, 107. macrofungi of wooded patches 63 hebeloma crustuliniforme (bull.) quél. – m/s; 1999: (s 2001, l&s 2002), 2000-2001, 2003-2004: 12, 58, 73, 100; h. mesophaeum (pers.) quél. – m/s; 2000-2007: 58, 60, 73; h. sacchariolens quél. – m/s; 1999: (b&k 2000, s 2001, l&s 2002); h. sordescens vesterh. – m/s; 1999: (b&k 2000); h. velutipes bruchet [= h. longicaudum (fr.) ss. lange.] – m/s; 1998-1999: (s 2001, l&s 2002). hemipholiota populnea (pers.) bon – s/w; 2004: 111, 112. heterobasidion annosum (fr.) bref. s.l. – p/w and s/w; 2000-2002: 25, 28, 35, 40, 41, 47, 49, 50. *hohenbuehelia cyphelliformis (berk.) o.k. mill. – s/l; 2001: 58; h. fluxilis (fr.) p.d. orton – s/w, r; 1997, 1999: (b&k 2000), 2000-2002: 2, 6, 29, 39, 42, 36; h. mastrucata (fr.) singer – s/w; 2001: 38, 43, 45. hydropus floccipes (fr.) singer – s/s; 2006: 100. hygrocybe conica (schaeff.) p. kumm. var. conica – m/s; 2000-2001, 2006: 6, 100; h. insipida (j.e. lange) m.m. moser – m/s, e; 2000-2001: 6. hygrophoropsis aurantiaca (wulfen) maire – s/w; 2000: (s 2001, l&s 2002), 2000, 2002-2003: 25, 26, 40, 41, 46, 49, 50, 52, 83, 87, 98, 107. hygrophorus hypothejus (fr.) fr. – m/s, i; 2004: 79. hymenochaete rubiginosa (dicks.) lév. – s/w; 2000-2001, 2003: 50, 86. hypholoma fasciculare (huds.) p. kumm. var. fasciculare – (rev. z. heinrich), s/w; 1997-1999: (b&k 2000, s 2001, l&s 2002), 2000-2005: 9, 11, 17, 21, 22, 25, 26, 28, 30, 35, 38, 39, 40, 41, 42, 43, 44, 45, 47, 49, 50, 67, 73, 82, 87, 89, 90, 97, 100, 102, 104, 114; h. lateritium (schaeff.) p. kumm. – (rev. z. heinrich), s/w; 20002003, 2006: 17, 25, 26, 28, 38, 45, 50, 60, 98, 114. infundibulicybe [clitocybe] gibba (pers.) harmaja (rev. h. komorowska) – s/s; 19982000: (s 2001, l&s 2002), 2000-2002: 38, 39, 50. inocybe auricoma (batsch) j.e. lange – m/s; 2000-2002, 2004, 2007: 12, 21, 39, 58; i. bongardii (weinm.) quél. – m/s; 2000: (s 2001, l&s 2002); i. calida velen. – m/s; 2000: 18; i. calospora quél. – (rev. a. ronikier), m/s, v; 2000-2002 (r 2005a); i. cincinata (fr.) quél. var. major (s. petersen) kuyper [= i obscura (pers.) gillet – m/s; 2007: 60; i. curvipes p. karst. – m/s; 1998-2000: (s 2001, l&s 2002), 2000-2005: 58, 60, 73; i. dulcamara (alb. & schwein.) p. kumm. – m/s; 2003: 107; i. erubescens a. blytt – m/s; 1999: (b&k 2000), 2001-2002, 2004: 34, 48, 63, 100; i. fuscidula velen. var. fuscidula – m/s; 2000-2001, 2004: 34, 114; i. geophylla (pers.) p. kumm. – m/s; 1997-1998: (b&k 2000), 2000-2001: 12, 20, 24, 100; i. grammata quél. & le bret. – m/s, v; 1999: (s 2001, l&s 2002); i. hirtella bres. – m/s; 2000-2001: 48; i. lacera (fr.) p. kumm. var. lacera – m/s; 2000, 2002, 2004: 11, 44, 45, 58, 73; i. lanuginosa (bull.) p. kumm. – m/s; 2000-2002: 28, 43, 50; i. lilacina (peck.) kauffman [= i. geophylla var. lilacina gillet] – m/s; 19971998: (b&k 2000); i. napipes j.e. lange – m/s; 1999-2000: (s 2001, l&s 2002), 2000: 7; i. perlata (cooke) sacc. – m/s; 1997-1999: (b&k 2000), 2000-2004: 8, 20, 34, 80, 100; i. petiginosa (fr.) gillet – m/s; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2001: 73; i. posterula (britzelm.) sacc. – m/s;2001, 2004, 2007: 58, 60 73; i. praetervisa quél. – m/s; 2000-2002: 31; i. pusio p. karst. – m/s; 2000-2001: 20, 28; i. rimosa (bull.) p. kumm. – m/s; 1998: (b&k 2000), 2000-2002: 29, 47, 48, 58, 100; i. squamata j.e. lange – m/s, r; 2000-2001: 29, 97. inonotus radiatus (sowerby) p. karst. – s/w; 2001: 23. 64 a. kujawa kuehneromyces mutabilis (schaeff.) singer & a.h. sm. – s/w; 1997-1999: (b&k 2000), 2000-2001, 2003-2004: 21, 43, 47, 67, 113. laccaria amethystina cooke – m/s; 1997-1999: (b&k 2000), 2000-2002: 11, 17, 43, 50; l. bicolor (maire) p.d. orton – m/s; 1997-1999: (b&k 2000, s 2001, l&s 2002), 2000: 45; l. laccata (scop.) berk. & broome – m/s; 1999-2000: (s 2001, l&s 2002), 2000-2005: 2, 5, 9, 11, 12, 17, 21, 25, 26, 28, 30, 33, 34, 35, 36, 39, 43, 44, 45, 46, 58, 73; l. proxima (boud.) pat. – m/s; 1998-2000: (s 2001, l&s 2002), 2003-2004: 73; l. tortilis (bolton) cooke – m/s; 1999-2000: (b&k 2000, s 2001, l&s 2002), 2000-2002, 2004: 39, 58, 73. lachnella alboviolascens (alb. & schwein.) fr. – s/w; 2005, 2007: 58, 74. lacrymaria lacrymabunda (bull.) pat. – s/s; 1998-1999: (b&k 2000), 2000, 20022004: 13, 100, 120. lactarius blennius (fr.) fr. – m/s; 1997-1999: (b&k 2000), 2000-2001: 17, 35, 48; l. camphoratus (bull.) fr. – m/s; 2000-2002: 43, 44, 45, 50, 66; l. chrysorrheus fr. – m/s, r; 2000-2001: 44, 50; l. deliciosus (l.) gray – m/s; 1996, 1998: (k&k 1997, s 2001, l&s 2002); l. fluens boud. – m/s; 2001: 5; l. glyciosmus (fr.) fr. – m/s; 1999: (s 2001, l&s 2002), 2001, 2004, 2006: 39, 73; l. necator (bull.) p. karst. – m/s; 1998: (s 2001, l&s 2002), 2000, 2004, 2006-2007: 39, 47, 73, 96; l. obscuratus (lasch) fr. – m/s; 2001: 36; l. omphaliformis romagn. – m/s; 2000-2002: 23, 29; l. pubescens (schrad.) fr. – m/s; 2007: 58; l. pyrogalus (bull.) fr. – m/s; 2001-2002: 35, 100; l. quietus (fr.) fr. – m/s; 1998, 2000: (s 2001, l&s 2002), 2000-2003: 2, 9, 22, 38, 40, 50, 66, 73, 83, 86, 100; l. rufus (scop.) fr. – m/s; 20002003: 28, 46, 114; l. tabidus fr. [= l. theiogalus (bull.) gray ss. neuhoff] – m/s; 2000-2003: 25, 26, 30, 43, 44, 45, 51, 67, 114; l. torminosus (schaeff.) pers. – m/s; 2001-2002: 58, 73. laetiporus sulphureus (bull.) bondartsev & singer – p/w; 1997, 1999: (b&k 2000), 2000-2006: 57, 60, 67, 68, 70, 80, 92, 94, 95, 97, 100, 115, 120. leccinum pseudoscabrum (kallenb.) šutara – m/s; 2001: 11; l. scabrum (bull.) gray – m/s; 2000, 2003, 2005, 2007: 73, 114. lentinus tigrinus (bull.) fr. – s/w, r; 2000, 2003: 96, 97. lepiota brunneoincarnata chodat & c. martín – s/s, v; 2000-2002: 8, 10, 13; l. castanea quél. – s/s; 2001, 2006: 21, 30, 39, 114; l. clypeolaria (bull.) p. kumm. – s/s; 20002001: 30, 39; l. cristata (bolton) p. kumm. – s/s; 1997: (b&k 2000), 2001-2004, 2006: 8, 12, 13, 58, 60, 100; l. echinella quél. & g.e. bernard [= l. setulosa j.e. lange] – s/s; 2006: 100; l. fuscovinacea f.h. møller & j.e. lange – s/s; 2003: 92; l. griseovirens maire [= l. pseudofelina j.e. lange] – s/s, e; 1998: (b&k 2000), 2001-2002, 2006: 27, 100; l. lilacea bres. – s/s, ex; 2000-2001, 2006: 12, 13, 18, 100; l. magnispora murrill – s/s; 2000, 2002, 2004: 23, 66, 97; l. subgracilis kühner – s/s; 2000: (s 2001, l&s 2002); l. subincarnata j.e. lange – s/s; 1998: (s 2001, l&s 2002), 2000-2002: 8, 12, 13, 20, 27. lepista flaccida (sowerby.) pat. – (rev. h. komorowska), s/l; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2002: 2, 3, 4, 5, 7, 27, 32, 38; l. nuda (bull.) cooke – s/s; 1998, 2000: (s 2001, l&s 2002), 2000-2002, 2004, 2006-2007: 4, 5, 23, 27, 35, 37, 38, 58, 73, 80, 100; l. saeva (fr.) p.d. orton – s/s; 2000-2002: 8, 13, 19, 20; l. sordida (schumach.) singer – s/s; 2000: 73. macrofungi of wooded patches 65 leucoagaricus leucothites (vittad.) wasser var. leucothites – s/s; 1998: (s 2001, l&s 2002), 2001, 2004, 2006: 58, 92, 93; *l. melanotrichus (malençon & bertault) trimbach – s/s; 2006: 100. lycoperdon lividum pers. – s/s; 2006-2007: 73; l. molle pers. – s/s; 2000-2002, 2005: 12, 39, 50, 60; l. nigrescens pers. – s/s; 2002, 2004: 49, 79; l. perlatum pers – s/s; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2007: 2, 5, 9, 12, 17, 24, 25, 28, 30, 32, 35, 37, 38, 39, 40, 41, 49, 50, 58, 73, 87, 100; l. pyriforme schaeff. – s/w; 1997, 2000: (b&k 2000, s 2001, l&s 2002), 2000-2004: 39, 42, 47, 50, 73; l. umbrinum pers. – s/s; 2000-2001: 11, 49, 50. lyophyllum fumosum (pers.) p.d. orton – s/s; 2000-2002: 48. *macrocystidia cucumis (pers.) joss. var. latifolia (j.e. lange) imazeki & hongo – s/s; 2000-2007: 7, 27, 58, 60, 73. macrolepiota mastoidea (fr.) singer – s/s; 2000: 32; m. procera (scop.) singer – s/s; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2004, 2006: 2, 3, 7, 16, 21, 27, 30, 37, 38, 39, 47, 58, 64, 66, 67, 73, 74, 79, 90, 95, 100, 107, 115, 122. macrotyphula fistulosa (holmsk.) r.h. petersen – s/w, r; 2000-2004: 8, 23, 29, 73, 112; m. juncea (fr.) berthier – s/l, r; 2000-2001: 23, 38, 50. marasmiellus vaillantii (pers.) singer – s/l; 2000-2003: 2, 9, 14, 18, 31, 36, 47, 50, 87. marasmius anomalus lasch var. anomalus – s/l; 2000: 3; m. collinus (scop.) singer – s/s; 2000: (s 2001, l&s 2002); m. curreyi berk. & broome – s/l; 2000-2007: 3, 29, 33, 37, 58, 60, 73, 82, 90, 105; *m. epiphylloides (rea) sacc. & trotter – s/l; 2005: 100; m. epiphyllus (pers.) fr. – s/l; 2000-2003, 2007: 6, 21, 23, 28, 29, 30, 31, 60, 69; m. oreades (bolton) fr. – s/s; 1997-2000: (b&k 2000, s 2001, l&s 2002), 20002007: 7, 8, 9, 12, 16, 20, 24, 27, 31, 33, 37, 51, 58, 62, 67, 68, 73, 78, 100, 105, 114, 120, 125; m. rotula (scop.) fr. – s/w; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2004, 2006: 1, 2, 4, 5, 6, 8, 12, 13, 14, 15, 18, 19, 27, 33, 35, 36, 62, 68, 73, 80, 86, 87, 100, 120, 125; m. setosus (sowerby) noordel. – s/l, v; 1997-1999: (b&k 2000), 2000-2001, 2003: 35, 36, 48, 100; m. torquescens quél. – s/l; 1999: (b&k 2000), 2000-2002: 1, 2, 5, 6, 13, 14, 15, 18, 35, 36; m. wynnei berk. & broome – s/s; 2000: 20. melanoleuca brevipes (bull.) pat. – s/s; 2000: (s 2001, l&s 2002), 2000, 2002-2005: 76, 100, 120; m. cognata (fr.) konrad & maubl. – s/s; 1998-1999: (b&k 2000), 2000: 92; m. excissa (fr.) singer var. excissa – s/s; 1999-2000: (s 2001, l&s 2002), 2000: 12; m. grammopodia (bull.) pat. – s/s; 2001: 58; m. melaleuca (pers.) murrill – s/s; 1998: (s 2001, l&s 2002); m. polioleuca (fr.) kühner f. polioleuca – s/s; 2000-2001: 37, 48, 58; *m. polioleuca (fr.) kühner f. pusilla boekhout & kuyper – s/s; 2001, 2004-2005: 12, 13, 16, 60, 66, 73. *melanomphalia nigrescens m.p. christ. – m?/s; 2003-2004, 2006: 100. melanophyllum haematospermum (bull.) kreisel – s/s, r; 2000-2001, 2003: 6, 12, 13, 92, 103. meripilus giganteus (pers.) p. karst. – p/w, p; 1999-2002: (b&k 2000, k 2003b); 20032006: 85, 100. mycena acicula (schaeff.) p. kumm. – s/l; 1997, 1999: (b&k 2000), 2000-2004, 2006: 1, 6, 10, 12, 13, 14, 15, 18, 19, 29, 30, 31, 34, 35, 36, 58, 80, 100, 114; m. adscendens (lasch) maas geest. – s/l, e; 2000-2002: 14, 27; m. aetites (fr.) quél. – s/l; 1997: (b&k 2000), 2000, 2002, 5, 30; m. amicta (fr.) quél. – s/l; 2001: 73; m. arcangeliana bres. – s/w; 1998: (b&k 2000); m. aurantiomarginata (fr.) quél. – s/l, v; 1999: 66 a. kujawa (b&k 2000); m. cinerella (p. karst.) p. karst. – s/l; 2000-2001, 2003: 11, 25, 28, 40, 41, 46, 50, 107; m. citrinomarginata gillet – s/l; 1999: (b&k 2000), 2002: 25; m. epipterygia (scop.) gray var. epipterygia – s/l; 2000-2002, 2004: 25, 26, 28, 30, 39, 40, 41, 46, 47, 50, 78; m. erubescens höhn. – s/w; 1998: (s 2001, l&s 2002), 2000-2002: 4, 5, 12, 18; m. filopes (bull.) p. kumm. – s/l; 2000: (s 2001, l&s 2002), 2001-2002, 2004: 14, 16, 23, 30, 36, 38, 47, 48, 73; m. flavescens velen. – s/l, r; 1999: (b&k 2000), 2001: 13; m. flavoalba (fr.) quél. – s/l; 2000-2001: 19, 26; m. galericulata (scop.) gray – s/w; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2006: 2, 5, 6, 12, 13, 15, 19, 21, 22, 23, 25, 26, 27, 28, 29, 30, 31, 32, 34, 37, 38, 40, 41, 42, 45, 46, 47, 48, 51, 60, 67, 68, 83, 97, 98, 100, 107, 114, 115; m. galopus var. candida j. e. lange ss. robich (2003) – s/l; 2001: 26; m. galopus (pers.) p. kumm. var. galopus ss. robich (2003) – s/l; 1997-1999: (b&k 2000, s 2001, l&s 2002), 2000-2003: 2, 5, 11, 12, 13, 15, 17, 22, 23, 25, 26, 28, 32, 35, 36, 38, 39, 40, 41, 42, 45, 46, 47, 48, 49, 50, 67, 68, 83, 114; m. galopus var. leucogala (cooke) j.e. lange ss. robich (2003) – s/l; 2000-2002: 23, 25, 39; m. haematopus (pers.) p. kumm. – s/w; 2000-2004: 23, 31, 60, 68, 96; m. inclinata (fr.) quél. – s/w; 2000: 6; m. leptocephala (pers.) gillet – s/l; 1998-1999: (b&k 2000), 20002003, 2006-2007: 1, 2, 5, 6, 7, 12, 14, 16, 21, 22, 23, 25, 27, 30, 32, 33, 35, 36, 38, 39, 40, 41, 42, 43, 47, 58, 73, 83, 114; m. leptophylla (peck) sacc. – s/w, r; 2000: 15; m. metata (fr.) p. kumm. – s/l; 2000: (s 2001, l&s 2002), 2000-2002: 25, 26, 28, 35, 46, 49; m. niveipes (murrill) – s/w; 1999: (s 2001, l&s 2002), 2001-2002: 23; m. olida bres. – s/w, v; 2001-2002: 12, 13, 14; m. olivaceomarginata (massee) massee – s/l; r; 2000-2002, 2006-2007: 9, 12, 16, 18, 20, 21, 23, 27, 31, 36, 47, 73; m. polyadelpha (lasch) kühner – s/l; 2000-2001: 2, 6, 9, 23; m. polygramma (bull.) gray – s/w; 1997, 1999: (b&k 2000, s 2001, l&s 2002), 2001-2002: 17, 36, 50, 100; m. pseudocorticola kühner – s/w, v; 1997, 1999: (b&k 2000), 2000: 13; m. pura (pers.) p. kumm. – s/s; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2004, 2006-2007: 1, 5, 7, 16, 17, 21, 22, 23, 25, 28, 30, 35, 36, 37, 38, 39, 40, 41, 42, 47, 51, 58, 73, 86, 97, 100; m. pura f. alba (gillet) kühner ss. robich (2003) – s/l; 2000-2001: 22, 23, 41; m. purpureofusca (peck) sacc. – s/w, v; 1997-1998: (b&k 2000), 2000-2002: 11, 25, 26, 28, 41, 43, 50, 114; m. rosea (schumach.) gramberg – s/s; 1998-1999: (s 2001, l&s 2002), 2005: 73; m. sanguinolenta (alb. & schwein.) p. kumm. – s/l; 1998-2000: (b&k 2000, s 2001, l&s 2002), 20002004, 2006-2007: 2, 4, 6, 11, 12, 16, 17, 21, 22, 23, 25, 26, 27, 28, 29, 30, 33, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 49, 50, 60, 67, 68, 73, 79, 82, 83, 86, 97, 98, 100, 114; m. speirea (fr.) gillet – s/l; 1997-1999: (b&k 2000), 2000-2004, 2006: 1, 4, 6, 8, 12, 13, 15, 18, 19, 23, 31, 34, 35, 36, 97, 100; m. stipata maas geest. & schwöbel – s/w; 1998: (b&k 2000), 2000-2003: 35, 60, 114; m. stylobates (pers.) p. kumm. – s/l; 1998: (s 2001, l&s 2002), 2000-2007: 6, 9, 27, 28, 33, 35, 36, 39, 40, 41, 46, 49, 50, 58, 73, 83, 82, 86, 114; m. tintinabulum (fr.) quél. – s/w; 1997, 1999: (b&k 2000), 2002, 2005, 2007: 100; m. vitilis (fr.) quél. – s/l; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2004, 2006-2007: 1, 2, 4, 5, 9, 11, 15, 18, 19, 21, 23, 27, 29, 30, 35, 36, 38, 44, 45, 47, 49, 50, 60, 67, 73, 80, 100; m. zephirus (fr.) p. kumm. – s/l; 1998-1999: (b&k 2000), 2000-2004: 11, 17, 25, 26, 28, 38, 40, 41, 46, 79, 107. mycenastrum corium (guers.) desv. – s/s, v; 2000, 2002-2004: (k et al. 2004), 20042007: 112. macrofungi of wooded patches 67 mycenella bryophila (voglino) singer – s/s; 2000-2001: 6, 13, 31, 34, 48. mycenitis [marasmius] scorodonius (fr.) wilson & desjardin – s/l; 2000-2003: 4, 5, 16, 17, 26, 30, 35, 39, 41, 43, 46, 47, 51, 68, 83, 83, 86, 93, 97, 114. myxomphalia maura (fr.) hora – s/s, i; 2002: 3. naucoria celluloderma p.d. orton – m/s; 1998: (b&k 2000), 2000, 2002: 29; n. escharioides (fr.) p. kumm. – m/s; 1997-1999: (b&k 2000), 2000-2002: 23, 29, 30, 36; n. scolecina (fr.) quél. – m/s; 1997-1999: (b&k 2000), 2000-2003: 23, 29, 97; n. subconspersa p.d. orton – m/s; 1998: (b&k 2000), 2000: 15, 23, 29. neolentinus lepideus (fr.) redhead & ginns – s/w; 2001-2003: 89, 99, 100, 107, 119; n. schaefferi (weinm.) redhead & gins [= lentinus cyathiformis (schaeff.) bres.] – s/w, e; 2000: 80. ossicaulis lignatilis (pers.) redhead & ginns – s/w, v; 1997: (b&k 2000), 2001-2002: 2, 18, 92. panaeolina foenisecii (pers.) maire – s/s; 1999: (s 2001, l&s 2002), 2000-2003, 2007: 1, 16, 33, 58, 60, 73, 82. panaeolus acuminatus (schaeff.) gillet – s/s; 2000-2001: 32; p. cinctulus (bolton) sacc. [= p. subbalteatus (berk. & broome) sacc.] – s/s; 2006: 112; p. fimicola (pers.) gillet – s/s; 2000: (s 2001, l&s 2002). panellus mitis (pers.) singer – s/w; 2003-2004: 79, 107. panus conchatus (bull.) fr. – s/w, r; 2004: 107. parasola [coprinus] auricoma (pat.) redhead, vilgalys & hopple – s/s; 1998-1999: b&k 2000, s 2001, l&s 2002, as: coprinus hemerobius), 2000-2004, 2006: 34, 75, 80, 97, 100, 120; p. plicatilis (curtis) redhead, vilgalys & hopple – s/s; 19981999: (b&k 2000, s 2001, l&s 2002), 2000-2003: 9, 10, 16, 34, 36, 58, 60, 100. paxillus filamentosus (scop.) fr. – m/s, 1997-1998: r; (b&k 2000), 2000-2001: 23; p. involutus (batsch) fr. – m/s; 1997-2000: (b&k 2000, s 2001, l&s 2002), 20002007: 8, 11, 15, 21, 23, 25, 26, 28, 30, 31, 35, 39, 40, 41, 43, 44, 45, 46, 47, 50, 51, 55, 58, 73, 79, 107, 114. phaeolus schweinitzii (fr.) pat. – p/w and sw; 2000-2002, 2004: 25, 67, 97. phaeomarasmius borealis rald – s/w; 2000, 2002: 47. phallus impudicus l. – m/s; 1999-2003 (k&k 1997, b&k 2000, s 2001, l&s 2002, k 2003b, ), 2000-2004: 47, 62, 66, 67, 83, 86, 96, 97, 98, 113, 114. phellinus pomaceus (pers.) maire – p/w; 2004: 100; ph. robustus (p. karst.) bourdot & galzin – p/w; 2004: 40. phlebia tremellosa (schrad.) burds. & nakasone – s/w; 1999: (b&k 2000), 2001, 2004: 29, 73. pholiota adiposa (batsch) p. kumm. – s/w; 1997: (b&k 2000, as: p. aurivella); ph. lenta (pers.) singer – s/w; 2000: (s 2001, l&s 2002), 2000-2001: 50; ph. squarrosa (weigel) p. kumm. – s/w; 1998: (b&k 2000), 2000-2002: 10, 13. pholiotina arrhenii (fr.) singer – s/s; 2001-2002: 1, 6, 13, 19; ph. [conocybe] mairei (watling) enderle – s/s; 1999: (b&k 2000); ph. vexans (p.d. orton) bon [= conocybe blattaria (fr.) kühner] – s/s; 2000-2001: 58. piptoporus betulinus (bull.) p. karst. – s/w; 1999: (b&k 2000), 2000-2003: 30, 51, 54, 62, 67, 76, 83, 100, 113. pleurotus dryinus (pers.) p. kumm. – s/w; 2003: 61, 107; p. ostreatus (jacq.) p. kumm. – s/w; 1997-1998, 2000: (b&k 2000, s 2001, l&s 2002), 2000, 2007: 8, 60. 68 a. kujawa pluteus atromarginatus (konrad) kühner – s/w; 2003: 124; p. cervinus (schaeff.) p. kumm. – s/w; 1997-1999: (b&k 2000, s 2001, l&s 2002), 2000-2004: 2, 11, 17, 23, 29, 30, 31, 35, 38, 39, 40, 41, 44, 45, 46, 47, 52, 63, 67, 69, 73, 80, 92, 96, 97, 98, 117; p. cinereofuscus j.e. lange – s/s; 2000: 6, 47; p. ephebeus (fr.) gillet – s/s, r; 2000, 2003-2004, 2006: 31, 34, 67, 100; p. hispidulus (fr.) gillet – s/w, r; 2000-2002: 29; p. insidiosus vellinga & schreurs – s/w; 2000-2002: 29, 31; p. nanus (pers.) p. kumm. – s/s; 2000-2002, 2004-2006: 1, 7, 10, 13, 20, 29, 30, 31, 39, 60, 67, 100, 114; p. nanus (pers.) p. kumm. f. griseopus (p.d. orton) vellinga ss. vellinga (1990) – s/s; 2000-2001: 9, 37; p. petasatus (fr.) gillet – s/w, r; 2000, 2002, 2004, 2007: 1, 89, 100, 104; p. phlebophorus (ditmar) p. kumm. – s/w; 2000: 100; p. plautus (weinm.) gillet ss vellinga (1990) – s/w, i; 1998: (b&k 2000), 20002003: 6, 11, 12, 13, 23, 29, 31, 39, 83, 86, 97; p. podospileus sacc. & cub. – s/w; 1999: (b&k 2000), 2001-2003, 2006-2007: 13, 29, 58, 97, 100; p. podospileus sacc. & cub. f. minutissimus (maire) vellinga ss vellinga (1990) – s/s i s/w; 20002002, 2004, 2006-2007: 6, 10, 15, 50, 58, 60, 100; p. romellii (britzelm) sacc. – s/w; 1999: (b&k 2000), 2000-2003, 2006-2007: 6, 18, 24, 34, 29, 31, 39, 73, 80, 97, 114; p. salicinus (pers.) p. kumm. – s/w; 2000: (s 2001, l&s 2002), 2000-2001, 2004: 29, 30, 67; p. thomsonii (berk. & broome) dennis – s/w; 2002: 31; p. umbrosus (pers.) p. kumm. var. umbrosus – s/w; 1997-1998: (b&k 2000). polyporus arcularius (batsch) fr. – s/w; 1998: (b&k 2000), 2000, 2002-2003: 11, 18, 25, 42, 44, 63, 73, 82, 90; p. brumalis (pers.) fr. – s/w; 2000: (s 2001, l&s 2002), 2000-2002, 2004, 2006-2007: 21, 23, 25, 41, 73, 107; p. ciliatus fr. – s/w; 2000-2001, 2003, 2006: 21 25, 51, 66, 83, 107; p. durus (timerm.) kreisel [= p. badius (pers.) schwien.] – s/w; 2000, 2003-2004: 39, 63, 80, 100; p. leptocephalus (jacq.) fr. [= p. varius (pers.) fr.] – s/w; 1997: (b&k 2000); p. squamosus (huds.) fr. – p/w; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2004, 2006: 1, 18, 63, 64, 70, 80, 92, 100, 120, 125. postia caesia (schrad.) p. karst. – s/w; 2000: 46; p. ptychogaster (f. ludw.) vesterh., knudsen & hansen – s/w, r; 2000-2001, 2003: 25, 41, 117; p. stiptica (pers.) jülich – s/w; 2000-2001, 2003: 46, 86; p. subcaesia (a. david) ryvarden & gilb. – s/w; 1997: (b&k 2000), 2001, 2003: 47, 62. psathyrella candolleana (fr.) maire – s/w; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2003, 2006: 4, 6, 7, 11, 19, 23, 29, 30, 37, 38, 39, 66, 73, 75, 83, 87, 97, 102, 114; p. canoceps (kauffman) a.h. sm. – s/s, e; 2000: (s 2001, l&s 2002); p. conopilea (fr.) ulbr. – s/s; 1997: (b&k 2000), 2003: 67, 100; p. corrugis (pers.) konrad & maubl. [= p. gracilis (fr.) quél.] – s/l; 1999: (b&k 2000), 2000-2003: 3, 6, 12, 14, 19, 31, 38, 100; p. gyroflexa (fr.) ss. m.m. moser (according to legon i in. 2005 nomen dubium) – s/l; 1999: (s 2001, l&s 2002); p. marcescibilis (britzelm.) singer – s/l; 1999: (s 2001, l&s 2002); p. obtusata (pers.) a.h. sm. – s/l; 2000: (s 2001, l&s 2002); p. piluliformis (bull.) p.d. orton – s/w; 1997-1999: (b&k 2000, s 2001, l&s 2002), 2000-2002: 50, 90; p. prona (fr.) gillet – s/s; 1997-1999: (b&k 2000, s 2001, l&s 2002), 2002-2003, 2007: 6, 100; p. pseudocasca (romagn.) kits van wav. – s/w; 1999: (s 2001, l&s 2002); p. spadiceogrisea (schaeff.) maire – s/s; 1998-1999: (b&k 2000, s 2001, l&s 2002), 2000-2002, 2004: 6, 20, 29, 30, 34, 100, 120; p. subnuda (p. karst.) a.h. sm. (according to legon et al. 2005 nomen dubium, according to wojewoda 2003 insufficiently known species) – s/l; 2000: (s 2001, l&s 2002); p. tephrophylla (romagn.) bon – s/s; 2000-2002: 6, 24. macrofungi of wooded patches 69 pseudoclitocybe expallens (pers.) m.m. moser [= p. obbata (fr.) singer] – (rev. h. komorowska), s/s; 2002-2005: 58, 60. pseudocraterellus undulatus (pers.) rauschert – m/s; 2001: 11. psilocybe crobulus (fr.) singer – s/w; 2000-2002: 2, 35, 36, 50, 58; p. inquilinus (fr.) bres. – s/l; 2000, 2002, 2007: 25, 37, 50, 58, 73; p. montana (pers.) p. kumm. – s/s, r; 2007: 105; p. [melanotus] phillipsi (berk. & broome) vellinga & noordel. – s/l, e; 2000: 25. pycnoporus cinnabarinus (jacq.) p. karst. – s/w, r; 2000, 2004-2005: 79, 90, 105. ramaria flaccida (fr.) ricken – s/l; 2007: 58, 60; r. stricta (pers.) quél. – s/w; 1008, 2000: (b&k 2000, s 2001, l&s 2002), 2000-2002: 37. resupinatus trichotis (pers.) singer – s/w; 200-2001, 2004: 9, 35, 37, 47, 67. rhodocollybia butyracea (bull.) lennox f. butyracea – m/s; 1997-1998: (b&k 2000), 2001: 26; r. butyracea (bull.) lennox f. asema (fr.) antonín, halling & noordel. – m?/s; 1998: (s 2001, l&s 2002), 2000-2004, 2006-2007: 2, 4, 5, 7, 22, 23, 25, 26, 28, 30, 36, 38, 40, 41, 42, 44, 45, 46, 48, 49, 50, 73, 75, 79, 100; r. maculata (alb. & schwein.) singer var. maculata – m?/s; 2000-2001, 2003: 26, 40, 51, 117. rhodocybe gemina (fr.) kuyper & noordel. – s/s; 2005-2007: 60, 114. rickenella fibula (bull.) raithelh – s/s, (b); 2000-2004, 2007: 6, 21, 22, 23, 25, 26, 26, 30, 35, 37, 38, 40, 41, 42, 46, 47, 49, 50, 58, 66, 68, 73, 82, 83, 98, 100, 105, 107, 114; r. swartzii (fr.) kuyper – s/s, (b); 2000-2004, 2007: 6, 23, 29, 35, 41, 46, 58, 67, 79. ripartites tricholoma (alb. & schwein.) p. karst. – s/l; 2001, 2003-2004, 2006-2007: 38, 73. rugosomyces [calocybe] carneus (bull.) bon – s/l; 2007: 58, 73. russula alnetorum romagn. – m/s, v; 1998: (b&k 2000), 2000-2001: 23; r. amoenolens romagn. – m/s, r; 2007: 73; r. claroflava grove – m/s; 2000-2003: 30, 98, 114; r. cyanoxantha (schaeff.) fr. – m/s; 1997-1998, 2000: (b&k 2000, s 2001, l&s 2002), 2000-2001: 100; r. delica fr. – m/s; 1997-1998: (b&k 2000), 2001: 48; r. emetica (schaeff.) pers. – m/s; 2000-2002: 44, 45; r. fellea (fr.) fr. – m/s; 20002001: 17, 48, 90; r. foetens (pers.) fr. – m/s; 2000: (s 2001, l&s 2002); r. fragilis (pers.) fr. – m/s; 2000-2002: 9, 44, 45, 50, 73; r. grisea (pers.) fr. – m/s; 1999: (b&k 2000); r. heterophylla (fr.) fr. – m/s; 2000-2002: 11; r. insignis quél. [= r. livescens (batsch) quél.] – m/s, i; 2000-2002: 8, 24, 33, 47; r. ionochlora romagn. – m/s; 2000-2001, 2003: 43, 47, 90; r. mairei singer – m/s; 1997-1999: (b&k 2000), 2000-2001: 48, 90; r. nigricans (bull.) fr. – m/s; 2000-2002: 11, 43, 45, 73; r. ochroleuca pers. – m/s; 2000-2002: 11, 43; r. risigallina (batsch) sacc. var. risigallina – m/s; 2000-2002: 73; r. solaris ferd. & winge – m/s; 1998: (b&k 2000); r. vesca fr. – m/s; 2002: 43, 47. sarcomyxa [panellus] serotina (schrad.). p. karst. – s/w; 2007: 100. schizophyllum [auriculariopsis] amplum (lév.) nakasone – s/w; 1997, 1999: (b&k 2000), 2000-2003: 12, 14, 15, 34, 39; s. commune fr. – s/w; 1998: (b&k 2000), 2000-2002, 2007: 24, 43, 45, 47, 100. scleroderma areolatum ehrenb. – m/s; 2000, 2006: 58, 73; s. bovista fr. [= s. fuscum (corda) e. fischer] – m/s; 2000-2003, 2006-2007: 21, 39, 47, 58, 73, 90, 100, 125; s. citrinum pers. – m/s; 2000-2001, 2003, 2005: 23, 47, 51, 62, 83, 96, 105, 107; s. verrucosum (bull.) pers. – m/s; 1997-1998: (b&k 2000), 2000-2001, 2003, 20052007: 9, 20, 21, 33, 36, 38, 55, 58, 73, 120, 125. 70 a. kujawa simocybe centunculus (fr.) singer var. centunculus – s/w; 2002: 6; s. haustellaris (fr.) watling – s/w; 1997: (b&k 2000), 2000-2003: 1, 4, 6, 7, 12, 13, 14, 15, 16, 18, 19, 20, 21, 27, 31, 34, 35, 36, 100. sparassis crispa (wulfen) fr. – p/w and s/w, p, r; 2000-2001: (k&k 1009, k 2003b), 2000-2003, 2006: 66, 73, 76, 89, 96, 114. sphaerobolus stellatus tode: pers. – s/w; 2000-2001, 2006: 2, 4, 7, 11, 13, 24, 25, 26, 35, 36, 42, 73. stereum gausapatum (fr.) fr. – s/w; 2000: 73; s. hirsutum (willd.) gray – s/w; 1998: (b&k 2000), 2000-2003, 2007: 3, 21, 25, 28, 30, 38, 40, 41, 42, 43, 44, 45, 49, 58, 73, 83, 86, 100; s. rugosum (pers.) fr. – s/w; 1999: (s 2001, l&s 2002), 2000-2003: 22, 25, 28, 30, 38, 40, 41, 42, 49, 96; s. subtomentosum pouzar – s/w; 1997: (b&k 2000), 2000-2003, 2005: 13, 21, 23, 29, 38, 47, 62, 85, 86, 96. strobilurus esculentus (wulfen) singer – s/l; 2000-2001, 2007: 26, 46, 100; s. stephanocystis (hora) singer – s/l; 2000-2001, 2003, 2006: 40, 49, 50, 83, 90, 106, 114; s. tenacellus (pers.) singer – s/l; 2000-2003: 40, 47, 49, 60, 73, 89, 90, 96. stropharia aeruginosa (curtis) quél. – (rev. z. heinrich), s/w i s/s; 1997-1998, 2000: (b&k 2000, s 2001, l&s 2002), 2000-2001, 2003: 25, 26, 28, 40, 41, 42, 46, 47; s. coronilla (bull.) quél. – (rev. z. heinrich), s/s; 1998: (b&k 2000), 2000-2007: 4, 9, 10, 24, 58, 60, 64, 73; s. cyanea (bull.) tuom. – (rev. z. heinrich), s/s; 19982000: (s 2001, l&s 2002), 2000-2007: 1, 2, 4, 5, 6, 7, 8, 22, 23, 27, 32, 36, 39, 42, 58, 60, 73; s. inuncta (fr.) quél. – (rev. z. heinrich), s/l; 2000-2002: 58, 73; s. melanosperma (pers.) gillet – (rev. z. heinrich), s/s, e; 2001-2002, 2004, 20062007: 58; s. pseudocyanea (desm.) morgan – (rev. z. heinrich), s/s, r; 2000: 24; s. rugosoannulata murrill – (rev. z. heinrich), s/s; 2000, 2004: 100, 108; s. semiglobata (batsch) quél. – s/l; 2000: (s 2001, l&s 2002). suillus bovinus (l.) roussel – m/s; 2005: 106; s. granulatus (l.) roussel – m/s; 2000: 40; s. grevillei (klotzsch) singer var. grevillei – m/s; 1998-2000: (s 2001, l&s 2002), 2000-2007: 17, 58, 90, 97, 99, 106, 107; s. luteus (l.) roussel – m/s; 20002001, 2004-2005: 58, 73, 82, 106; s. viscidus (l.) roussel – m/s; 2000-2001, 2004: 48, 58. *syzygospora mycetophila (peck) ginns – p/f; 2001: 46. tapinella atrotomentosa (batsch) šutara – s/w; 1998: (b&k 2000), 2000-2001, 2003, 2006: 26, 48, 51, 66, 106, 114; t. panuoides (fr.) e.-j. gilbert f. panuoides – s/w; 2001: 66, 81. thelephora caryophyllea (schaeff.) pers. – m/s, v; 2005-2007: 60; t. penicillata fr. – s/l, v; 2006: 73; t. terrestris ehrh. – m/s; 1998-2000: (s 2001, l&s 2002), 20002001, 2004, 2006: 25, 26, 28, 41, 43, 44, 46, 58, 73. trametes hirsuta (fr.) pilát – s/w; 2002-2005, 2007: 31, 58, 96, 100, 106; t. versicolor (l.) pilát – s/w; 1999: (s 2001, l&s 2002), 2000-2003, 2007: 14, 30, 31, 43, 44, 45, 49, 62, 96, 100, 114. trichaptum abietinum (pers.) ryvarden – s/w; 2003: 52, 83, 114. tricholoma argyraceum (bull.) p. kumm. – m/s; 2000: 20; t. inamoenum (fr.) gillet – m/s; 1999: (s 2001, l&s 2002); t. populinum j.e. lange – m/s; v; 2004: 112; t. scalpturatum (fr.) quél. – m/s; 2000-2001, 2007: 48, 58; t. sulphureum (bull.) p. kumm. – m/s; 1999: (b&k 2000), 2001: 50; t. terreum (schaeff.) p. kumm. – m/s; 1999: (b&k 2000), 24, 100. tricholomopsis rutilans (schaeff.) singer – s/w; 2000-2002: 25, 49, 73. macrofungi of wooded patches 71 tubaria dispersa (pers.) singer – s/l; 2000-2004: 6, 8, 12, 13, 20, 27, 34, 100; t. furfuracea (pers.) gillet [= t. hiemalis bon] – s/l; 1999-2000 (s 2001, l&s 2002, as: t. furfuracea et t. hiemalis), 2000-2003, 2005: 2, 4, 5, 9, 13, 14, 16, 17, 22, 36, 42, 48, 58, 63, 67, 92, 114; t. pellucida (bull.) fr. (according to legon et al. 2005 it is synonym of t. furfuracea or t. romagnesiana) – s/l; 1999: (s 2001, l&s 2002); t. romagnesiana arnolds – s/l; 2000: (s 2001, l&s 2002). tylopilus felleus (bull.) p. karst. – m/s; 2000-2001, 2003: 26, 44, 45, 114. typhula erythropus (pers.) fr. – s/l; 1999: (b&k 2000), 2001: 23, 29; t. incarnata lasch – s/l; 2005: 58; t. setipes (grev.) berthier – s/l; 2001-2002: 7, 100. vascellum pratense (pers.) kreisel – s/s; 2001-2007: 58, 73, 62, 114. volvariella gloiocephala (dc.) boekhout & enderle – s/s; 1999-2000: (s 2001, l&s 2002), 2000-2001, 2004, 2007: 58, 93, 73, 99, 112; v. murinella (quél.) courtec. – s/s, r; 2000-2002, 2004, 2006: 13, 15, 100, 107; v. taylori (berk.) singer – s/s, r; 1999: (b&k 2000); v. volvacea (bull.) singer – s/w, v; 1999: (b&k 2000). xerocomus badius (fr.) j.e. gilbert – m/s; 1997, 1999: (b&k 2000), 2000-2004, 2006: 26, 28, 30, 43, 44, 45, 46, 50, 67, 68, 73, 79, 107, 113; x. chrysenteron (bull.) quél. – m/s; 1997-2000: (b&k 2000, s 2001, l&s 2002), 2000-2007: 2, 5, 7, 9, 11, 17, 20, 22, 24, 28, 30, 32, 33, 35, 36, 38, 40, 41, 43, 44, 47, 49, 52, 60, 67, 73, 74, 80, 98, 100, 114, 125; x. rubellus (krombh.) quél. – m/s; 1998-2000: (s 2001, l&s 2002), 2000-2005, 2007: 7, 18, 20, 21, 36, 47, 66, 67, 73, 80, 100, 107, 114; x. subtomentosus (l.) quél. – m/s; 1998: (b&k 2000, s 2001, l&s 2002), 2000-2001, 2003: 11, 28, 107, 113. xeromphalina cornui (quél.) j. favre – s/l; 2000-2003: 40, 82, 83, 98, 114. xerula radicata (relhan) dörfelt – s/w; 1997, 1999-2002: (b&k 2000, r 5005b), 2002003: 35, 36. final remarks species richness of mycobiota and diversity of functional groups of macrofungi in the general dezydery chłapowski landscape park are similar to those observed in many areas where habitats are less transformed and forest cover is higher. even though wooded areas cover only 15% of the park and are strongly transformed by human activity and include forest monocultures, the area is very important for conservation of the fungal species that have an ability to adapt to anthropogenic habitats. they include some species that are new to poland, very rare or regarded as extinct. the role of the patchy structure of farmland for conservation of fungi will be presented and analyzed in my next articles. acknowledgements. i sincerely thank all those who helped me, but particularly prof. anna bujakiewicz for guidance during my research, access to literature in this field, and discussions. i am also grateful for valuable comments to prof. władysław wojewoda and prof. stefan friedrich. special thanks are due to dr zofia heinrich for verification of identification of stropharia and hypholoma spp.; to dr halina komorowska for verification of identification of clitocybe, pseudoomphalina, and gammundia spp.; to dr andrzej brzeg for phytosociological consultations; and to dr hab. halina ratyńska for access to the unpublished map of potential vegetation of the park. i thank also sylwia ufnalska, m.sc., for correction of the english manuscript. 72 a. kujawa references adamczyk j. 1997. grzyby makroskopowe bezodpływowych zagłębień śródpolnych południowej części kujaw. przegl. przyr. 8 (1/2): 131–134. agerer r. 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(in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. 3 ed. w. szafer institute of botany, polish academy of sciences, kraków: 53-70. wojterski t., wojterska h., wojterski m. 1981. potencjalna roślinność naturalna środkowej wielkopolski. bad. fizjogr. pol. zach. b, 32: 7–35. woś a., tamulewicz j. 1996. the microclimate of the turew region. (in:) l. ryszkowski, n.r. french, a. kędziora (eds). dynamics of an agricultural landscape. pwril: 37–44. zając a. 1978. założenia metodyczne “atlasu rozmieszczenia roślin naczyniowych w polsce”. wiad. bot. 22: 145–155. zajączkowski k. 2005. regionalizacja potrzeb zadrzewieniowych w polsce. ibl. rozprawy i monogra-monografie 4. macrofungi of wooded patches 75 grzyby wielkowocnikowe zadrzewień śródpolnych w krajobrazie rolniczym. i. różnorodność gatunkowa streszczenie powyższy artykuł jest pierwszym z serii czterech artykułów prezentujących wyniki badań nad grzybami wielkoowocnikowymi przeprowadzonych w krajobrazie rolniczym wielkopolski. w latach 2000-2007 badano różnorodność gatunkową grzybów wielkoowocnikowych zadrzewień śródpolnych, parków wiejskich i lasów gospodarczych w parku krajobrazowym im. gen. dezyderego chłapowskiego. przez trzy lata badania prowadzono na 50 stałych powierzchniach. przez cały okres badań zbierano też grzyby poza stałymi powierzchniami. stwierdzono występowanie 569 taksonów macromycetes, a po uwzględnieniu nielicznych danych z literatury liczba znanych taksonów z tego terenu wynosi 615. w pierwszym artykule zawarto opis terenu badań oraz wykaz gatunków i ich lokalizację. kolejne artykuły będą dotyczyły: gatunków chronionych, rzadkich oraz stwierdzonych w polsce po raz pierwszy,• roli zbiorowisk drzewiastych w ochronie różnorodności gatunkowej grzybów wielkoowoc-• nikowych w krajobrazie rolniczym, zmian w różnorodności gatunkowej i strukturze zbiorowisk grzybów w zbiorowiskach le-• śnych będących pod wpływem silnej antropopresji. fig. 1. general dezydery chłapowski landscape park. fig. 2. distribution of permanent plots in the general dezydery chłapowski landscape park in relation to potential vegetation (after ratyńska and szwed unpublished data, modified). 2014-01-01t11:48:49+0100 polish botanical society xerophiles and other fungi associated with cereal baby foods locally produced in uganda mady a. ismail1,2, hannington k. taligoola2 and rebecca nakamya2 1department of botany, faculty of science, p. o. box 71516, assiut university, assiut, egypt, madyismail@yahoo.com 2department of botany, faculty of science, makerere university p. o. box 7062, kampala, uganda ismail m.a., taligoola h.k., nakamya r.: xerophiles and other fungi associated with cereal baby foods locally produced in uganda. acta mycol. 47 (1): 75–89, 2012. fifty samples from five baby food products mainly made of cereal flour(s) were analyzed. the moisture contents of these products were between 11.14% and 11.9%, a level below 14.0%, the recommended level for safe storage of cereal grains and their products. the mycological analysis was carried out using the dilution plate method and two isolation media (dg18 for isolation of xerophilic fungi and drbc for fungi in general). a total of 80 species related to 37 genera in addition to some unidentified fungal and yeast species were recorded on both media from the five products. the products were contaminated abundantly by xerophilic fungi which were occurring in 88% of food samples and accounting for 18.1% of the total cfu as recorded on dg18. the highest contamination level by xerophiles was registered in mwebaza rice porridge (a component of rice flour) and the lowest in mukuza (a product of maize, soyabean and sorghum flours). 11 xerophilic species were recorded of which aspergillus and eurotium (4 species each) were the predominant giving rise to 9.1% and 8.9% of the total cfu, with a. wentii, a. candidus, e. cristatum and e. repens were the most contaminating species. of the fungi recorded other than xerophiles, species of aspergillus (particularly a. flavus followed by a. niger), penicillium (p. citrinum, p. oxalicum), fusarium (f. solani, f. tricinctum), cladosporium (c. sphaerospermum) and yeasts were the most predominant. contamination of such foods is a matter of health hazard as these foods are for babies. so, the use of fresh, well-dried and uncontaminated flours for production of such foods is recommended. key words: mycobiota, spoilage, contamination, food products, dg18, drbc acta mycologica vol. 47 (1): 75–89 2012 76 m.a. ismail et al. introduction baby foods rich in carbohydrates and proteins are being produced from dried cereal and leguminous grains/seeds. the manufacture of baby foods involves mixing the cereal flour with some additional ingredients such as powders of soya beans, dried fish or fruits. since these dried foods possessed low moisture content levels, they are subject to contamination and spoilage by microorganisms. fungi probably contaminate and spoil more foods than any other group of microorganisms. they render contaminated foods not only unpalatable, but also unsafe for consumption by producing mycotoxins (munimbazi, bullerman 1996). sanchis et al. (1982) noted that certain species if humidity reaches a sufficient level would start producing toxic metabolites. the presence of aflatoxins in foods is of great concern in terms of food safety since they are among the most active ingested carcinogens (pitt, hocking 2009). spoilage of such foods is due to a group of fungi termed as xerophiles that are capable of rapid growth above 0.77 aw and of slow growth at 0.75 aw and below-down to about 0.68 aw. taligoola et al. (2004); pitt, hocking (2009) stated that the most common causes of spoilage of dried cereals are species of eurotium particularly e. chevalieri, e. repens, e. rubrum and e. amstelodami l. mangin. a variety of yeasts are also common on cereal grains and flour (kurtzman et al. 1970; aran, eke 1987; pitt, hocking 2009). other ingredients such as powders of soya beans, milk, dried fish and fruits were also found to be contaminated with a wide variety of fungal species (mislivec, bruce 1977; sutic et al. 1979; bullerman 1979; jarchovska et al. 1980; ito, abu 1985; pitt et al. 1994; ismail, saad 1997; pitt, hocking 2009). many reports have been published earlier at different localities of the world on the microbiological quality of baby foods (jarchovská et al. 1980; moustafa et al. 1984; abdel-sater, ismail 1993; zohri et al. 1995; munimbazi, bullerman 1996) or their ingredients such as wheat (aran, eke 1987; mills et al. 1995), milk powder (ismail, saad 1997), rice (pitt et al. 1994; taligoola et al. 2004, 2010), maize (ismail et al. 2003), sorghum (pitt et al. 1994), millet (mishra, daradhiyar 1991), corn chips, breakfast cereal (bullerman, tsai 1994; zohri et al. 1995), and baby foods imported into uganda (ismail et al. 2008, 2010). in uganda, knowledge about fungi contaminating locally produced foods is needed. henceforth, this work was designed to survey the xerophilic and other fungi associated with baby foods locally manufactured in uganda. materials and methods fifty samples of five baby food products manufactured locally (10 packets each) were randomly collected from different shops at five towns of uganda (kampala, jinja, mbarara, masaka and mbale). most of these products are made in these towns. each of these foods contained at least one or more cereal flour. the names, components and the producing companies of these products are shown in table 1. determination of moisture content. three sub-samples of 50 g each were taken from each food sample and put in aluminium foil dish. these were dried in an oven xerophiles and other fungi 77 at 110°c for 24 hours, and reweighed (magan, lacey 1985; pitt, hocking 2009). the moisture content of each sample was expressed as the average percentage of the weight loss of the three replicates. mycological analysis. all food samples were analyzed mycologically on two isolation media: (1) dichloran 18% glycerol agar (dg18) which contains glycerol and glucose needed for the growth of xerophilic fungi (hocking, pitt 1980) and (2) dichloran rose bengal chloramphenicol agar, drbc (king et al. 1979). dilutions were prepared by shaking 10 g of each sample in 90 ml diluent of 0.1% peptone water (kurtzman et al. 1971). serial tenfold dilutions were prepared and 1 ml aliquots of the appropriate dilution were placed in sterile petri dishes. eight plates were used for each food sample (4 plates for each isolation medium) i.e. 80 plates for each type of food products and in total 400 plates for the fifty food samples. plates were incubated at room temperature (25-27°c) in day and night cycle of light conditions for 10-14 days for dg18 and for 10 days for drbc plates. the growing fungi were enumerated, isolated and identified. identification of fungi. the identification of different fungal groups was carried out based on their macroscopic and microscopic features using the methods and keys described by raper, fennell (1965) for aspergillus and its teleomorphs; booth (1971) and leslie, summerell (2006) for species of fusarium; pitt (1979) for species of penicillium; ellis (1971), moubasher (1993), domsch et al. (2007), and pitt & hocking (2009) for other fungi. statistical analysis. data were subjected to analysis of variance (anova), using the statistical analysis system, (sas institute inc., 1996). means were compared with l.s.d. test at p ≤ 0.05 levels. results and discussion each of the five baby foods investigated contained at least one or more cereal products (tab. 1). moisture content of the products investigated. all samples of the five baby food products were characterized by the average moisture contents ranging from 11.14 % table 1 food products investigated, their ingredients, producing companies and their mean moisture contents no product ingredients producing company mean moisture content (n=10) 1 baby soya maize flour, soya bean flour, carrot flour east african basic food, ltd 11.47±0.23 2 kayebe maize flour, powdered enkejje (haplochromis fish), soya bean flour kayebe sauce packers, ltd 11.14±0.38 3 mwebaza rice porridge rice flour ebenezer packers, ltd 11.9±0.23 4 jacinta millet flour millet flour mahimba company, ltd 11.6±0.09 5 mukuza maize flour, sorghum flour, soya bean flour hodeco, ltd 11.52±0.09 78 m.a. ismail et al. ± 0.38 % in kayebe to 11.9 ± 0.23 % in mwebeza rice porridge (tab. 1). however, all the products had moisture contents below 14.0%, the recommended level for safe storage of cereal grains and their products (christensen, kaufman 1974; taligoola et al. 2004, 2010; ismail et al. 2003, 2008, 2010). incidence of xerophilic fungi in baby food products (recovered mainly on dg18%). baby foods locally manufactured in uganda were abundantly contaminated by xerophilic fungi which occurred in 88% of food samples and accounted for 18.08% of the total cfu as recovered on dg18 (tab. 2). the contamination level with xerophiles ranged from 2.62% of the total cfu in mukuza to 60.44% in mwebaza rice porridge (tab. 3). these high contamination levels with xerophiles indicate that the baby foods may have stayed in shops and supermarkets for a long time where they might have been invaded by storage fungi. bullerman (1979) reported that, food stuffs in shops and markets are actually under storage, hence fungal contamination is likely to occur. ten xerophilic species belonging to four genera were recovered from the five products. aspergillus (72% of 50 food samples) and eurotium (64%) were isolated in high incidences while wallemia johan-olsen and xeromyces l. r. fraser were infrequent (tab. 2). the levels of contamination by xerophilic fungi in locally produced foods (88% of the food samples) were higher than those reported by ismail et al. (2010) in imported ones (54%), however similar species of the genera aspergillus, eurotium and wallemia were reported from both foods though in different frequencies. eurotium (4 species) occurred in 64% of the food samples and constituted the majority of xerophilic cfu (48.89%) from the 5 products. the highest level of contamination with eurotium species was found in mwebaza rice porridge (24.56% of the total cfu), and the lowest in mukuza (1.58%) (tab. 3). eurotium species have been reported on cereal baby foods imported into uganda (ismail et al. 2010), maize, rice, soya beans and dried fish (el-kady, youssef 1993; pitt et al. 1994; taligoola et al. 2004). among the eurotium species reported, e. cristatum and e. repens were the most predominant, accounting for 21.17% and 17.98% of the total xerophiles cfu. this finding agrees with earlier reports by kurata et al. (1968) who found these two species to be common spoilage fungi in many cereals (rice, maize, sorghum, wheat and barley) in storage. the observation of e. repens in high occurrence on the foods analysed is inconsistent with the findings whereby e. repens and other unidentified eurotium species were recovered in rare frequency on turkish cereals and cereal products (aran, eke 1987). e. chevalieri and e. rubrum were infrequently encountered, however, these two species were reported earlier as frequent on maize and rice (kurata et al. 1968; taligoola et al. 2004) and on baby foods imported to uganda (ismail et al. 2010). xerophilic aspergilli (4 species) accounted for 50.13% of the xerophiles cfu and 9.06% of the total fungi cfu (tab. 2). they were recovered in 72% of food samples from the five products. the highest level of contamination was found in mwebaza rice porridge (35.87% of the total cfu) and the lowest was registered in mukuza (1.04%). a. wentii was the most prevalent species, accounting for 25.95% of the xerophiles cfu. it was found most heavily contaminating kayebe and baby soya (major components: maize and soyabeans). in this respect, low levels of infection by a. wentii were recorded on soya beans, maize, paddy rice and sorghum in thailand and indonesia (pitt et al. 1994) and on maize (ismail et al. 2003) and rice (taligoola xerophiles and other fungi 79 ta bl e 2 in ci de nc e of fu ng i i n lo ca l b ab y fo od p ro du ct s on d ic hl or an 1 8% g ly ce ro l a ga r (d g 18 ) an d di ch lo ra n ro se b en ga l c hl or am ph en ic ol a ga r (d r b c ) ta xa d g 18 d r b c c f u c f u % f % o r so ur ce c f u c f u % f % o r so ur ce x er op hi lic fu ng i 28 22 5 18 .0 8 88 h 1, 2, 3, 4, 5 18 25 1. 08 25 m 1, 2, 3, 5 a sp er gi llu s (t ot al ) 14 15 0 9. 06 72 h 1, 2, 3, 4, 5 12 50 0. 74 20 l a . c an di du s l in k 65 50 4. 20 16 l 1 15 0 0. 09 6 r 5 a . p en ic ill io id es s pe ga zz in i 17 5 0. 11 8 r 1, 3, 5 50 0. 03 4 r 1, 2 re st ri ct us s m it h 10 0 0. 06 8 r 1, 2, 3 75 0. 04 4 r 1, 2 a . w en tii w eh m er 73 25 4. 71 72 h 1, 2, 3, 4, 5 97 5 0. 58 14 l 2 e ur ot iu m ( to ta l) 13 80 0 8. 87 64 h 1, 2, 3, 4, 5 50 0 0. 30 10 r 1, 2, 3 e . c he va lie ri m an gi n 24 75 1. 59 18 l 2, 3, 4, 5 20 0 0. 10 6 r 3 e . c ri st at um ( r ap er & f en ne ll) m al lo ch & c ai n 59 75 3. 84 56 h 1, 2, 3, 4, 5 30 0 0. 20 4 r 1, 2 e . r ep en s de b ar y 50 75 3. 26 34 m 1, 2, 3, 4, 5 e . r ub ru m k on ig , s pi ec ke rm an n & b re m er 27 5 0. 18 8 r 1, 3 po ly pa ec ilu m p is ce a . d . h oc ki ng & p it t 25 0. 01 4 2 r 1 w al le m ia s eb i ( fr ie s) v on a rx 15 0 0. 09 4 r 1 50 0. 03 2 r 1 x er om yc es b is po ru s l . r . f ra se r 12 5 0. 08 2 r 1 x er ot ol er an t f un gi 12 79 00 81 .9 2 10 0 h 1, 2, 3, 4, 5 16 74 15 98 .9 2 10 0 h 1, 2, 3, 4, 5 a cr em on iu m s tr ic tu m w . g am s 25 0. 02 2 r 5 20 0 0. 12 10 r 1, 3 a cr op hi al op ho ra s p. 75 25 4. 4 4 r 3 a lte rn ar ia a lte rn at e (f ri es ) k ei ss le r 75 0. 04 2 r 2 a sp er gi llu s (t ot al ) 32 70 0 20 .9 5 82 h 1, 2, 3, 4, 5 39 01 0 23 .0 5 84 h 1, 2, 3, 4, 5 a . a eg yp tia cu s m ou ba sh er & m ou st af a 50 0. 03 2 r 1 a . c ar bo na ri us ( b ai ni er ) t ho m 12 5 0. 08 2 r 1 75 0. 04 2 r 1 a . d efl ec tu s fe nn el l & r ap er 25 0. 01 4 2 r 1 a . fl av us l in k 23 72 5 15 .1 9 64 h 1, 2, 3, 4, 5 30 43 5 18 .0 74 h 1, 2, 3, 4, 5 a . f um ig at es f re se ni us 10 00 0. 64 20 l 1 97 5 0. 5 16 l 2, 3 a . n ig er v an t ie gh em 15 75 1. 01 42 m 1, 2, 3, 4, 5 18 00 1. 1 32 m 1, 2, 3, 4, 5 a . n om iu s k ur tz m an , h or n & h es se lt in e 25 0. 02 2 r 1 a . o ch ra ce us w ilh el m 80 0 0. 51 30 m 1, 2. 3, 4 45 0 0. 3 12 r 2, 3, 5 a . o ry za e (a hl bu rg ) c oh n 18 00 1. 15 22 l 2, 3, 4 52 5 0. 3 14 l l ,2 ,3 a . p ar as iti cu s sp ea re 50 0. 03 4 r 2 22 5 0. 13 10 r 1, 2 a . p ho en ic is ( c or da ) t ho m 25 0. 01 4 2 r 3 a . s yd ow ii (b ai ni er & s ar to ry ) t ho m & c hu rc h 72 5 0. 46 12 r 1, 3, 5 37 5 0. 2 8 r 2, 3 a . t am ar i k it a 20 75 1. 33 22 l 2, 3, 4 30 50 1. 8 24 l 1, 2, 3 a . t er re us t ho m 12 5 0. 08 6 r 1, 3 55 0 0. 3 8 r 2, 3 a . v er si co lo r ( v ui lle m in ) t ir ab os ch i 67 5 0. 43 22 l 1, 2, 3, 5 45 0 0. 3 18 l 1, 3 80 m.a. ismail et al. b ot ry ot ri ch um p ilu lif er um s ac ca rd o & m ar ch al 50 0. 03 2 r 1 b ys so ch la m ys fu lv a s . l . o lli vr & g . m . s m it h 52 5 0. 34 6 r 3 c ha et om iu m s p. 25 0. 01 4 2 r 1, 5 c la do sp or iu m ( to ta l) 61 75 3. 95 68 h 1, 2, 3, 4, 5 26 55 1. 6 46 m 1, 2, 3, 5 c . c la do sp or io id es ( fr es en iu s) d e v ri es 37 00 2. 37 34 m 1, 2, 3, 5 42 5 0. 14 8 r 1, 3, 5 c . h er ba ru m ( pe rs oo n) l in k 25 0. 02 2 r 1 c . s ph ae ro sp er m um p en zi g 24 50 1. 57 40 m 1, 3, 4, 5 22 30 1. 3 40 m 1, 2, 3, 4, 5 c oc hl io bo lu s lu na tu s r . n el so n & h aa si s 50 0. 03 4 r 1 c ur vu la ri a sp . 25 0. 01 4 2 r 3 d or at om yc es s p. 25 0. 01 4 2 r 3 e m er ic el la n id ul an s (e id am ) v ui lle m in 15 0 0. 09 2 r 3 e pi co cc um n ig ru m l in k 25 0. 01 4 2 r 1 e up en ic ill iu m s p. 15 0 0. 09 8 r 2, 5 fe nn el lia fl av ip es w ile y & s im m on s 25 0. 01 4 2 r 1 fu sa ri el la s p. 50 0. 03 2 r 1 fu sa ri um ( to ta l) 13 55 0 8. 68 42 m 1, 2, 3, 4, 5 43 30 0 25 .6 56 h 1, 2, 3, 4, 5 f. e qu is et i ( c or da ) sa cc ar do 50 0. 03 4 r 4 f. la te ri tiu m n ee s 31 25 2. 0 16 l 4 f. o xy sp or um s ch le ch te nd al 25 0. 02 2 r 1 f. p oa e (p ec k) w ol le nw . 25 0. 02 2 r 3 f. s ol an i ( m ar ti us ) sa cc ar do 25 0. 02 2 r 4 24 10 0 14 .2 5 26 m 1, 2, 3, 4, 5 f. tr ic in ct um ( c or da ) sa cc ar do 98 50 6. 31 12 r 5 17 77 5 10 .5 20 l 1, 5 f. v er tic ill io id es ( sa cc ar do ) n ir en be rg 42 5 0. 02 10 r 2, 3 13 75 0. 8 32 m 1, 3, 4, 5 fu sa ri um s p. 75 0. 05 2 r 3 g eo tr ic hu m c an di du m l in k 25 0. 01 4 2 r 5 h yp om yc es c hr ys os pe rm us t ul . 75 0. 04 4 r 1 l as io di pl od ia th eo br om ae ( pa t.) g ri ff on & m au bl . 50 0. 03 2 r 1 m ic ro do ch iu m n iv al e (f ri es ) sa m ue ls & h al le tt 15 0 0. 09 6 r 5 12 5 0. 07 6 r 1, 3 m on ili a sp . 20 0 0. 1 2 r 1 m uc or ( to ta l) 42 75 2. 5 20 l 1, 2, 3, 5 m . p lu m be us b on or d. 19 50 1. 2 12 r 1, 2, 3, 5 m . r ac em os us f re se ni us 20 0 0. 13 8 r 1, 3 15 0 0. 09 6 r 2, 3 m uc or s p. 21 75 1. 3 14 l 2, 3 n eo sr to ry a fis ch er ii (w eh m er ) m al lo ch & c ai n 10 0 0. 06 2 r 3 43 25 2. 6 2 r 3 n eu ro sp or a cr as sa ( sh ea r & d od ge ) v. a rx 23 47 5 15 .0 4 30 m 3, 4, 5 81 75 4. 8 28 m 2, 3, 4, 5 pa ec ilo m yc es v ar io tii b ai ni er 40 0 0. 26 10 r 1, 3 35 0 0. 2 12 l 1, 2, 3 ta bl e 2 – co nt . xerophiles and other fungi 81 pe ni ci lli um ( to ta l) 16 62 5 10 .6 5 80 h 1, 2, 3, 4, 5 13 40 0 7. 9 58 h 1, 2, 3, 4, 5 p. c he rm es in um b io ur ge 15 0 0. 09 6 r 5 p. c hr ys og en um t ho m 65 0 0. 42 8 r 2 p. c itr in um t ho m 40 25 2. 58 28 m 1, 2, 3, 5 59 50 3. 5 40 m 1, 2, 3, 4, 5 p. c or yl op hi lu m d ie rc kx 75 25 4. 82 18 l 2, 3, 4, 5 25 0 0. 15 8 r 1, 2, 3 p. is la nd ic um s op p 50 0. 03 2 r 3 p. o xa lic um c ur ri e & t ho m 41 25 2. 64 44 m 1, 2, 3, 4, 5 60 75 3. 50 24 l 2, 3, 4 p. p in op hi lu m h ed gc oc k 75 0. 04 6 r 3 p. p ub er ul um b ai ni er 25 0. 01 4 2 r 3 p. p ur pu ro ge nu m s to ll 50 0. 03 2 r 1 p. v ar ia bi le s op p 75 0. 04 4 r 1, 3 p. v ir id ic at um w es tl in g 95 0 0. 5 16 l 1, 2 pe ni ci lli um s p. 50 0. 03 2 r 3 p ho m a sp . 32 5 0. 2 2 r 1 r hi zo pu s st ol on ife r ( e hr en be rg ) v ui lle m in 41 00 2. 63 22 l 3, 4, 5 50 75 3. 0 36 m 1, 3, 4, 5 sc op ul ar io ps is c an di da ( g ue gu en ) v ui lle m in 50 0. 03 2 r 1 t he rm oa sc us a ur an tia cu s m ie he 25 0. 01 4 2 r 2 tr ic ho de rm a ha rz ia nu m r if ai 50 0. 03 4 r 1, 2 o th er u ni de nt ifi ed fu ng i 12 50 0. 74 18 l 1, 3, 5 y ea st s 29 37 5 18 .8 1 22 l 1, 2, 3 35 77 5 21 .2 34 m 1, 2, 3, 4, 5 r ho do to ru la m uc ila gi no sa ( jö rg en se n) h ar ri so n 25 0 0. 16 2 r 2 97 5 0. 6 20 l 2, 3 y ea st s (b ro w n) 12 5 0. 07 4 r 1, 4 y ea st s ( w hi te ) 29 12 5 18 .6 5 22 l 1, 2, 3, 5 34 75 0 20 .5 14 l 3, 4, 5 y ea st s (y el lo w ) 25 0. 01 4 2 r 3 to ta l f un gi 15 61 25 10 0 10 0 h 1, 2, 3, 4, 5 16 92 40 10 0 10 0 h 1, 2, 3, 4, 5 n o. o f g en er a (3 7) 16 36 n o. o f s pe ci es ( 80 ) 48 65 a bb re vi at io ns : c f u = c ol on y fo rm in g un it s (c al cu la te d /g b ab y fo od p ro du ct in 5 0 sa m pl es ); c f u % = p er ce nt ag e co lo ny fo rm in g un it s (c al cu la te d pe r to ta l f un ga l c f u ); f % = p er ce nt ag e fr eq ue nc y (c al cu la te d pe r 50 s am pl es in ve st ig at ed ); o r = o cc ur re nc e re m ar ks ; h ig h (h ) = 5 010 0 % , m od er at e (m ) = 2 549 % , l ow ( l ) = 13 -2 4 % , r ar e (r ) = le ss th an 1 3 % ; s ou rc e: 1 = b ab y so ya , 2 = k ay eb e, 3 = m w eb az a ri ce p or ri dg e, 4 = j ac in ta m ill et fl ou r, 5 = m uk uz a. 82 m.a. ismail et al. ta bl e 3 c ol on y fo rm in g un it s an d pe rc en ta ge fr eq ue nc y of m os t c om m on fu ng i, an d th e nu m be r of g en er a an d sp ec ie s re co ve re d fr om th e fiv e ba by fo od pr od uc ts o n d g 18 a nd d r b c pr od uc t b ab y so ya k ay eb e m w eb az a ri ce p or ri dg e ja ci nt a m ill et fl ou r m uk uz a. m ed iu m d g 18 d r b c d g 18 d r b c d g 18 d r b c d g 18 d r b c d g 18 d r b c c om m on fu ng i c f u f % c f u f % c f u f % c f u f % c f u f % c f u f % c f u f % c f u f % c f u f % c f u f % x er op hi le s 15 00 70 37 5 50 92 75 10 0 11 25 50 11 20 0 10 0 20 0 30 51 75 90 10 75 80 10 0 20 e ur ot iu m cr is ta tu m 37 5 70 25 10 29 25 60 27 5 10 21 50 50 30 0 40 22 5 60 e . r ep en s 37 5 70 12 00 30 27 5 40 31 00 20 12 5 20 a sp er gi llu s w en tii 32 5 60 15 0 30 50 25 10 0 82 5 40 15 0 40 15 25 80 30 0 80 x er ot ol er an ts 29 00 10 0 45 05 10 0 35 20 0 10 0 47 91 0 10 0 73 25 10 0 21 40 0 10 0 42 00 0 10 0 47 27 5 10 0 39 97 5 10 0 46 35 0 10 0 a . fl av us 27 5 50 40 0 60 21 07 5 10 0 26 66 0 10 0 40 0 70 17 50 70 11 25 40 57 5 70 85 0 60 10 50 70 a . n ig er 12 5 20 22 5 40 52 5 70 70 0 60 17 5 30 80 0 30 62 5 60 25 10 12 5 30 50 20 a . o ch ra ce us 10 0 30 25 0 50 75 20 37 5 50 35 0 30 75 20 25 10 c la do sp or iu m cl ad os po rio id es 62 5 40 50 10 27 50 60 25 0 60 35 0 20 75 10 25 10 c . s ph ae ro spe rm um 27 5 30 80 5 80 75 40 60 0 40 50 0 30 11 00 60 55 0 10 47 5 70 30 0 40 fu sa ri um so la ni 10 0 20 25 10 23 92 5 10 0 75 10 f. v er tic illio id es 32 5 30 12 5 10 20 0 10 30 0 40 35 0 60 27 5 20 22 5 40 n eu ro sp or a cr as sa 75 10 30 0 30 10 0 20 23 05 0 10 0 78 25 10 0 12 5 20 17 5 10 pe ni ci lli um ci tr in um 10 0 20 25 0 40 28 50 50 41 25 80 57 5 30 12 50 30 17 5 40 50 0 40 15 0 10 p. o xa lic um 75 20 47 5 50 58 75 90 90 0 30 15 0 10 26 25 10 0 50 20 50 20 r hi zo pu s st ol on ife r 25 10 32 5 40 30 0 40 37 25 50 45 50 90 50 20 15 0 40 y ea st s (t ot al ) 75 10 25 10 60 0 50 35 0 50 12 5 10 72 5 20 34 07 5 40 c f u s of a ll fu ng i 44 00 10 0 48 80 10 0 44 47 5 10 0 49 03 5 10 0 18 52 5 10 0 21 60 0 10 0 47 17 5 10 0 47 27 5 10 0 41 05 0 10 0 46 45 0 10 0 n o. o f g en er a 8 24 4 11 12 18 7 6 7 8 n o. o f s pe ci es 25 39 19 27 35 34 16 10 20 16 fo r ab br ev ia ti on s ee t ab le 2 ; a = p ro du ct , b = m ed iu m , c = c om m on fu ng i; a *b *c l .s .d a t p ≤ 0 .0 5 = 2 10 .5 2 xerophiles and other fungi 83 et al. 2004) in uganda. a. candidus was the second most common xerophilic aspergillus species. it was recovered in 16% of the samples, however giving rise to high percentages of cfu (23.21% of xerophiles) and found only in baby soya. a. penicillioides and a. restrictus occurred infrequently in baby soya, kayebe, mwebaza rice porridge or mukuza. these two species were also uncommon in maize and rice in uganda (ismail et al. 2003; taligoola et al. 2004), and in baby foods imported to uganda (ismail et al. 2010), while a. restrictus was predominant on cereal grains in iran (lacey 1988) and a. penicillioides was isolated in high frequency from soya beans in thailand (pitt et al. 1994) and dried fish in indonesia (wheeler et al. 1986). the presence of a. restrictus on baby soya and kayebe (products of soya beans), is in agreement with the finding where a. restrictus was found invading soya beans whose moisture contents were between 12.5%-13% (christensen, kaufmann 1965). this fungus was first reported in stored grains by tuite, christensen (1955) and since then has been found to be a common cause of deterioration in all kinds of stored grains and seeds (christensen, kaufmann 1965). wallemia sebi and xeromyces bisporus were infrequently encountered and only from baby soya. w. sebi was earlier reported in maize and soya beans from thailand, and on peanuts, maize, paddy and milled rice, and soya beans from indonesia (pitt et al. 1994) and on pakistani and ugandan rice (taligoola et al. 2004), while x. bisporus was reported from dried prunes, spice powders, nutmegs, dates, fruit cakes and cookies (pitt, hocking 2009). it is worthy to mention that 7 of these xerophilic species in addition to polypaecilum pisce were reported on drbc but infrequently, accounting for a minor proportion of total cfu (1.08%). incidence of fungi other than xerophiles in baby food products (recovered on both dg18 and drbc). apart from the 11 xerophilic species, baby food products yielded a total of 33 genera and 69 species of xerotolerant fungi on both dg18 (38 species and 12 genera) and dichloran rose bengal chloramphenicol agar (drbc) (57 species belonging to 32 genera). the broadest spectrum of species was recorded in mwebaza rice porridge (35 species on dg18) and in baby soya (39 species on drbc), while the narrowest was recorded in jacinta millet flour (16 species on dg18 and 10 species on drbc). the current results revealed that the diversity of fungi was higher in locally produced foods (36 genera and 65 species on drbc) than in imported ones (21 and 42) analysed by ismail et al. (2008). aspergillus, penicillium, fusarium and cladosporium were the most predominant genera on both isolation media (tab. 2). penicillium and aspergillus were reported earlier as the most commonly isolated genera on starches (potatoes, rice, maize and wheat) intended for human consumption (suarez et al. 1981), on wheat, maize, sorghum and barley in egypt (moubasher et al. 1972; el-maghraby 1989). in a study on baby foods imported into uganda, the most common fungal genera were found to be similar to those reported in the current study from the local foods, however, species of aspergillus and penicillium were more dominant in locally produced foods, while species of cladosporium and fusarium were more common in imported ones (ismail et al. 2008). aspergillus was the most frequent genus. it emerged from 82% and 84% of food samples accounting for 20.95% and 23.05% of the total cfu on dg18 and drbc respectively. it was represented by 15 species of which a. flavus was the most common. a. flavus accouned for 15.19% and 18.0% of the total fungi cfu on dg18 and 84 m.a. ismail et al. drbc, respectively (tab. 2). it was recovered in high frequency from all products on both media but had its highest level of contamination in kayebe (tab. 3). the high incidence of a. flavus on kayebe whose major components are maize, soya beans and fish flours, is in agreement with earlier reports on maize (el-maghraby 1989; sebunya, yourtee 1990, munimbazi, bullerman 1996, ismail et al. 2003), on dried fish (ito, abu 1985) and on soya beans (el-kady, youssef 1993). contrary to the above findings, a. flavus was reported with low occurrence in maize and sorghum in egypt (el-kady et al. 1982) and soya beans in uganda (sebunya, yourtee 1990). a. niger came second and was found contaminating 42% and 32% of food samples from the five products, accounting for 1.01% and 1.1% of the total fungi cfu on dg18 and drbc, respectively. it was recovered in moderate frequency and most heavily contaminating kayebe and baby soya (products of maize and soya beans). this is in agreement with the findings of suarez et al. (1981), who recovered a. niger from starches intended for human consumption, where rice and maize were among these starches. also, sanchis et al. (1982) fond that a. niger caused severe deterioration to corn and sorghum. other four aspergillus species were recovered in low frequency of occurrence on both media, accounting collectively for 3.55% and 4.75% of the total fungi cfu on dg18 and drbc media, respectively, and these were: a. fumigatus, a. oryzae, a. tamarii and a. versicolor (tab. 2). earlier reports mentioned these species to occur less commonly on freshly ground mouldy maize meal (marasas, smalley 1972). a. ochraceus was reported moderately on dg18 but rarely on drbc giving rise to 0.51% and 0.3% of the total cfu respectively. on the other hand, 8 species of aspergillus were isolated in rare incidence on one or both media: a. aegyptiacus, a. carbonarius, a. deflectus, a. nomius, a. parasiticus, a. phoenicis, a. sydowii and a. terreus (tab. 2). a. carbonarius and a. parasiticus have been reported as rare species in food products imported into uganda (ismail et al. 2008, 2010), though some of the above species have been reported to be common on barley e.g., a. sydowii, a. ochraceus, a. terreus (moubasher et al. 1972) and a. terreus on maize (ismail et al. 2003), paddy rice (abdel-hafez et al. 1987) and flour (augustine et al. 1984). penicillium was the second most frequent genus, recovered from 80% and 58% of the total food samples on dg18 and drbc respectively, accounting for 10.65% and 7.9% of the total cfu. the highest levels of contamination with penicillia were recorded in maize flour-containing products (kayebe and baby soya), and mwebaza rice porridge and the least was found in mukuza (tabs 2 and 3). the above findings agree with earlier observation where penicillium was among the most common genera recovered from corn snacks (zohri et al. 1995) and starches (suarez et al. 1981). similarly, corn was found to be highly contaminated with penicillium species, while rice was found to be penicillia free (munimbazi, bullerman 1996). in contrast to our finding, whereby mukuza (a product of sorghum) had a low contamination level with penicillium species, diener et al. (1981) found penicillium to be among the most dominant genera on sorghum. p. citrinum (2.58% and 3.5%) and p. oxalicum (2.64% and 3.5% of the total cfu on dg18 and drbc respectively), the most common species in the present study, have been reported earlier to occur on starches (suarez et al. 1981), and barley (abdel-kader et al. 1979). p. oxalicum was also one of the chief species isolated from unstored corn kernels (mislivic, tuite 1970) and in preharvest corn from valencia, spain (jimenez et al. 1985). p. citrinum, xerophiles and other fungi 85 a nephrotoxigenic fungus, is known as citrinin-producer (mislivec, tuite 1970; frisvad 1983). p. viridicatum, a well known nephrotoxigenic species (frisvad 1983) and p. corylophilum were also isolated though in low frequency, respectively on drbc and dg18, constituting 0.5% and 4.82% of the total cfu. p. viridicatum was earlier recovered from starches intended for human consumption (suarez et al. 1981). carlton et al. (1968) found that p. viridicatum, p. oxalicum and p. multicolor grig.-manoil. & porad, all isolated from corn kernels in indiana were toxic to mice when included in their diets. other 7 penicillium species: p. chermesinum, p. chrysogenum, p. islandicum, p. pinophilum, p. puberulum, p. purpurogenum and p. variabile were rarely isolated from only one or two products (tab. 2). of these, p. variabile has been reported to cause severe deterioration in wheat, corn and sorghum (moubasher et al. 1972) and p. aurantiogriseum, p. chrysogenum, p. corylophilum, p. citrinum, p. expansum, p. islandicum, p. oxalicum, p. verrucosum, p. viridicatum from baby foods imported into uganda (ismail et al. 2008, 2010). fusarium occupied the third place with regard to its frequency, occurring in 42% and 56% of the samples on dg18 and drbc respectively. however its count (25.6% of the total cfu) was more than that of aspergillus and penicillium on drbc while lower than that of both genera on dg18 (tab. 2).this result agrees with the earlier finding that species of fusarium are field fungi (moubasher et al. 1972; christensen 1987) less tolerating the low water activity medium, dg18. the results revealed that fusarium represented the highest and the major food-contaminant. among the five products investigated, jacinta millet flour and mukuza were the most heavily contaminated having fusarium cfu in all their samples. it is possible that fusaria infected the cereals while still in the field and persisted even after the cereals were processed and stored. of eight species encountered, f. solani (0.02% and 14.24% of the total cfu) and f. tricinctum (6.31% and 10.5%) were the most contaminating on both dg18 and drbc respectively, though these were recovered in rare to moderate frequency. f. solani was found most heavily contaminating jacinta millet flour while f. tricinctum in mukuza (a product of sorghum and maize). this finding disagreed with earlier reports where f. solani and f. tricinctum were absent on millet and sorghum (diener et al. 1981; munimbazi, bullerman 1996). f. verticillioides constituted low percentages of the total cfu and was found most heavily contaminating baby soya and mwebaza rice porridge (products of maize and rice flour, respectively). f. verticillioides and f. tricinctum were registered earlier in maize meal (marasas, smalley 1972), maize stalks and grains (logrieco et al. 1988), and sorghum (diener et al. 1981). f. verticillioides is a major producer of moniliformin and fumonisins toxins that cause liver cancer in rats and oesophageal cancer in humans (lacey 1988, logrieco et al. 1988; sydenham et al. 1990). the remaining fusarium species were recorded infrequently either on dg18 (f. lateritium from 8 food samples from jacineta millet flour, f. oxysporum from one baby soya sample, f. poae from one sample of mukuza and unidentified fusarium species from one sample of mwebaza) or on drbc (f. equiseti from only 2 samples of baby soya). cladosporium (3 species) was also isolated from all products in high frequency (68% of food samples) on dg18 and moderate frequency (46%) on drbc, constituting 3.95% and 1.6% of the total cfu on dg18 and drbc, respectively. c. sphaerospermum was moderately isolated on both media with 1.57% and 1.3% of the total cfu while c. cladosporioides was moderate on dg18 and rare on drbc 86 m.a. ismail et al. (tab. 2). c. herbarum was rare and recovered only from 1 sample of kayebe. in this respect, mazen et al. (1984) found c. sphaerospermum in low frequency on maize. neurospora crassa, rhizopus stolonifer and yeasts (with rhodotorula mucilaginosa being the most common) were all isolated in moderate or low frequency on dg18 and in moderate frequency on drbc. these species were reported earlier as food spoilage fungi (pitt, hocking 2009). r. stolonifer had also been reported earlier from soya beans (el-kady, youssef 1993) and barley (abdel-kader et al. 1979). some other fungal species were isolated infrequently either in low or rare frequency from one or more food products on dg18 or drbc or both (tab. 2). analysis of variance. analysis of variance was computed on baby food products using anova test at 5% significance level (tab. 3). the calculated value of ftest = 8.84 at df 4. this is greater than the tabulated value fcritical = 1.96316, hence there is a significant difference in the total count of the different species recovered from the different food products on drbc and dg18. the type and the cfu of most fungal species recovered on drbc and dg18 from food products are probably dependent on the type of that product. this may also be due to the different ingredients including cereal flours involved in these products. conclusions the current results revealed that kayebe (a product of maize, fish and soya bean) and jacinta millet flour (a product of millet) were the most heavily contaminated by fungi cfu of the five products investigated, while baby soya was the lowest as determined on both isolation media. however, the highest contamination level by xerophiles was registered in mwebaza rice porridge and the lowest in mukuza. among eleven xerophilic species recorded on these baby foods, species of aspergillus and eurotium were the most common. in addition, a high incidence of aspergillus flavus, yeasts, fusarium solani, f. tricinctum, penicillium citrinum, p. corylophilum, p. oxalicum and cladosporium sphaerospermum on one or both isolation media were also recorded. many of these fungi are capable of producing mycotoxins. contamination of such foods (especially those for babies) is a matter of health hazard for human consumption. however their safety can be insured and improved greatly by using quality raw materials. as contamination occurs for cereal grains before, during or after harvesting, during drying process, or even during food production and this contamination could also be due to long-term storage, marketing under non-hygienic conditions of the food products. we suggest that monitoring fungal contaminations as well as mycotoxins should be carried out periodically and procedures to prevent mould contamination should be developed. acknowledgements. the authors are indebted to prof. bukenya-ziraba, the head of botany department, makerere university, kampala for the facilities he provided during this research. grateful acknowledgement is due to the egyptian fund for technical cooperation with africa for sponsoring prof. m. a. ismail at makerere university, giving him the opportunity to act as a supervisor of mrs. rebbecca nakamya. xerophiles and other fungi 87 references abdel-hafez s.i.i., el-kady i.a., mazen m.b., el-maghraby o.m. 1987. mycoflora and trichothecene toxins of paddy grains from egypt. mycopathologia 100 (2): 103–112. abdel-kader m.i.a., moubasher a.h., abdel-hafez s.i.i. 1979. survey of the mycoflora of barley grains in egypt. mycopathologia 69 (3): 143–147. abdel-sater m.a., ismail m.a. 1993. enzymatic studies on fungi associated with biscuits in egypt. international biodeterioration and biodegradation 31 (4): 277–292. aran n., eke d. 1987. mould mycoflora of some turkish cereals and cereal products. mircen journal of applied microbiology and biotechnology 3 (3): 281–287. augustine b., parvathi m., kalyanasundaram i. 1984. potential mycotoxigens in wheat and wheat products. journal of food science and technology, india 21 (5): 312–316. booth c. 1971. the genus fusarium. commonwealth mycological institute, kew, surrey, england, pp. 237. bullerman l.b. 1979. significance of mycotoxins to food safety and human health. journal of food protection 42 (1): 65–86. bullerman l.b., tsai w.y.j. 1994. incidence and levels of fusarium moniliforme, fusarium proliferatum and fumonisins in corn and corn-based foods and feeds. journal of food protection 57 (6): 541–546. carlton w.w., tuite j., mislivec p. 1968. investigations of the toxic effects in mice of certain species of penicillium. toxicology and applied pharmacolology 13 (3): 372–387. christensen c.m. 1987. field and storage fungi. in: food and beverage mycology, 2nd edition, beuchat l.r. 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a. 1986. fungi associated with indonesian dried fish. food microbiololgy 3 (4): 351–357. zohri a.a., abdel-sater m.a., ismail m.a. 1995. incidence of aflatoxins and mould flora in corn snacks. journal of food science and technology, india 32 (4): 289–294. 2014-01-02t12:04:10+0100 polish botanical society acaulospora rehmii and gigaspora margarita, arbuscular mycorrhizal fungi (glomeromycota) new for europe and poland, respectively janusz błaszkowski, sławomir kowalczyk and beata czerniawska department of plant pathology, university of agriculture słowackiego 17, pl 71 434 szczecin, jblaszkowski@agro.ar.szczecin.pl b ł a s z k o w s k i j . , k o w a l c z y k s . , c z e r n i a w s k a b .: acaulospora rehmii and gigaspora margarita, arbuscular mycorrhizal fungi (glomeromycota) new for europe and poland, respectively. acta mycol. 41 (1): 41 48, 2006. morphological characters of spores of acaulospora rehmii and gigaspora margarita (glomeromycota) were described and illustrated. spores of the two species were found in field collected mixtures of rhizosphere soil and roots collected in poland. attempts to produce spores in trap cultures succeeded only with g. margarita. all attempts to establish one species cultures of the two fungi failed. gigaspora margarita was for the first time found in poland and this paper is the first report of the occurrence of a. rehmii in europe. the known distribution of the two fungal species in the world is also presented. key words: arbuscular fungi, glomeromycota, mycorrhizae, distribution introduction continuing over 20-year studies of the occurrence of arbuscular mycorrhizal fungi of the phylum glomeromycota, spores of acaulospora rehmii sieverd. et s. toro and gigaspora margarita w.n. becker et i.r. hall, species earlier not recorded in europe and poland, respectively, were found. the aims of this paper are to describe and illustrate morphological characters of spores of a. rehmii and g. margarita and to present the distribution of these fungi in poland and the other regions of the world. materials and methods establishment of trap cultures and one-species cultures. rhizosphere soils and roots of sampled plants were collected from a depth of 5-30 cm using a small garden shovel. in the laboratory, about 100-g subsamples were taken from each sample to isolate spores of arbuscular mycorrhizal fungal species sporulating in the field. the remaining part of the sample was either air dried for 2 weeks and subsequently refrigerated at 4oc or directly used to establish trap cultures. trap cultures were estab acta mycologica vol. 41 (1): 41-48 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 42 j. błaszkowski et al. lished to obtain a great number of living spores and to initiate sporulation of species that were present but not sporulating in the field collections. the growing substrate of the trap cultures was the field-collected material mixed with an autoclaved coarsegrained sand coming from maritime dunes adjacent to świnoujście (ph 6.7; 12 and 26 mg l -1 p and k, respectively; b ł a s z k o w s k i 1995). the mixtures were placed in 9 x12.5-cm plastic pots (500 cm3) and densely seeded with plantago lanceolata l. plants were grown in a greenhouse at 15-30oc with supplemental 8-16-h lighting provided by one son-t agro sodium lamp (philips lighting poland s. a.) placed 1 m above pots. the maximum light intensity was 180 μe m-2s-1 at pot level. plants were watered 2-3 times a week. no fertilizer was applied during the growing period. trap cultures were harvested five to seven months after plant emergence. spores were extracted by wet sieving and decanting (g e r d e m a n n , n i c o l s o n 1963). presence of mycorrhizae was determined following clearing and staining of roots (p h i l l i p s , h a y m a n 1970) modified as follows: tissue acidification with 20% hcl instead of 1%, and trypan blue concentration 0.1% instead of 0.05% (k o s k e, pers. comm.). single-species pot cultures were established from about 10-20 spores stored before inoculation in water at 4oc for 24 h. after removal of soils debris, spores were collected in a pipette and transferred onto a compact layer of mycorrhizae-free roots of 10-14 day old seedlings of p. lanceolata placed at the bottom of a hole ca. 1 cm wide and 4 cm deep formed in a wetted growing medium filling 8-cm plastic pots (250 cm3). the growing medium was an autoclaved sand of maritime dunes adjacent to świnoujście with chemical properties listed above. subsequently, the spores were covered with another layer of roots of 4-6 additional p. lanceolata seedlings, and the roots and sandwiched spores were buried in the growing medium. finally, three to six seeds of p. lanceolata were placed on the surface of the growing substrate and covered with a thin layer of autoclaved sand. the cultures were harvested after 4-8 months. microscopy survey. morphological properties of spores and their subcellular structures were determined based on at least 20 spores mounted in polyvinyl alcohol/lactic acid/glycerol (pvlg; o m a r , b o l l a n , h e a t h e r 1979) and a mixture of pvlg and melzer’s reagent (1:1, v/v). spores were crushed to varying degrees by applying pressure to the coverslip and then stored at 65oc for 24 h to clear their contents of oil droplets. these were examined under an olympus bx 50 compound microscope equipped with nomarski differential interference contrast optics. microphotographs were recorded on a sony 3cdd colour video camera coupled to the microscope. terminology of spore structure is that suggested by s t ü r m e r & m o r t o n (1997) and wa l k e r (1983, 1986). spore colour was examined under a dissecting microscope on fresh specimens immersed in water. colour names are from k o r n e r u p & wa n s c h e r (1983). nomenclature of fungi and plants is that of wa l k e r & tr a p p e (1993) and m i r e k et al. (1995), respectively. the authors of the fungal names are as those presented at the url web page http://www.indexfungorum. org/authorsoffungalnames.htm. specimens were mounted in pvlg on slides and deposited in the department of plant pathology (dpp). colour microphotographs of spores and mycorrhizae of a. rehmii and g. margarita can be viewed at the url http://www.agro.ar.szczecin.pl/~jblaszkowski/. acaulospora rehmii and gigaspora margarita 43 descriptions of the species acaulospora rehmii sieverd. et s. toro spores single in the soil; develop laterally on the neck of a sporiferous saccule. spores dull yellow (3b3) to nankeen yellow (3b7); globose to subglobose; (90-)140(-165) μm diam. subcellular structure of spores consists of a spore wall and two inner germination walls (fig. 1). spore wall composed of three layers (layers 1-3). layer 1 evanescent, hyaline, (0.8-)1.0(-1.2) μm thick (fig. 2), continuous with the wall of a sporiferous saccule, rarely present in mature spores. layer 2 laminate, dull yellow (3b3) to nankeen yellow (3b7), (4.5-)10.0(-12.0) μm thick, ornamented with labyrinthine folds, 1.0-3.5 μm wide and 1.0-4.5 μm high when seen in a plane view and a cross view, respectively (figs 1-3). layer 3 semi-flexible, hyaline, (0.5-)0.7(-0.8) μm thick, rarely separable from layer 2 (fig. 2). germination wall 1 consists of two tightly adherent hyaline, semi-flexible layers, each 0.5-0.8 μm thick; these layers are tightly adherent to each other and, hence, difficult to see (figs 1, 2, 4, 5). germination wall 2 composed of two adherent layers (layers 1 and 2; figs 1 and 2). layer 1 flexible, hyaline, (0.5-)0.7(-1.0) μm thick, covered with small, <0.5 μm diam, granules scattering in crushed spores (figs 4 and 5). layer 2 flexible, hyaline, (0.6-)1.1(-1.4) μm thick in pvlg; its staining reaction in melzer’s reagent was not examined. germination orb not observed. sporiferous saccule hyaline; globose to subglobose; 90-150 μm diam, with a 1-layered wall, 0.8-1.7 μm thick; neck 50-80 μm long, tapering from 15.0-22.0 μm wide at the saccule to 12.0-19.0 μm wide at the spore attachment. saccule usually collapses and is detached in mature spores. cicatrix remaining after sporiferous saccule detachment is a slightly raised, circular scar, 8.0-11.5 μm diam (fig. 6). the unique character of spores of a. rehmii is the labyrinthine ornamentation of their structural laminate layer. the properties of the other layers of both the spore wall and the inner germination walls are similar to those of most the other species of the genus acaulospora. acaulospora rehmii is one of the nine recognized species of the genus acaulospora producing spores with an ornamented laminate structural layer. when observed under a dissecting microscope, spores of a. rehmii are dull because of the ornamented structural laminate layer and, thereby, most resemble in size and colour spores of a. bireticulata f.m. rothwell et trappe, a. dendiculata sieverd. et s. toro, a. lacunosa j.b. morton, a. spinosa c. walker et trappe, and a. tuberculata janos et trappe, fungi also producing ornamented spores. however, examination of crushed spores of a. rehmii under a compound microscope reveals them to be ornamented with labyrinthine processes (fig. 3), and those of a. bireticulata with polygonal reticulum with isolated projections at polygon centres (b ł a s z k o w s k i 2003; m o r t o n 2002; r o t h w e l l , tr a p p e 1979), a. denticulata with stubby knobs (m o r t o n 2002; s i e v e r d i n g , to r o 1987), a. lacunosa with round, prolate or irregularly-shaped pits (b ł a s z k o w s k i 2003; m o r t o n 1986, 2002), a. spinosa with closely packed rounded spines (wa l k e r , tr a p p e 1981), and a. tuberculata with polygonal spines or tubercles (j a n o s , tr a p p e 1982; m o r t o n 2002). other species of acaulospora forming spores with an ornamented outer surface of their laminate structural layer are a. foveata trappe and janos, a. paulinae błaszk., a. scrobiculata trappe, and a. undulata sieverd. compared with spores of 44 j. błaszkowski et al. a. rehmii, those of a. foveata are much greater [(240-)289(-300) vs. (82-)141(-175) μm diam in a. rehmii] and darker-coloured (red-orange to dark red-brown vs. light yellow to orange-brown), and spores of a. paulinae, a. scrobiculata, and a. undulata are lighter (hyaline to straw-coloured; b ł a s z k o w s k i 2003; m o r t o n 2002; s i e v e r d i n g , to r o 1987). additionally, spores of the latter two species are much smaller (55-92 μm diam; b ł a s z k o w s k i 2003; s i e v e r d i n g 1988). finally, spores of all the species compared here are ornamented with pits, and not with labyrinthine processes as those of a. rehmii. another important difference between a. rehmii and a. bireticulata, a. foveata, and a. tuberculata resides in the inner layer of the second germination wall of their spores. while this layer is thin and flexible in a. rehmii (figs. 4 and 5), that of a. bireticulata, a. foveata, and a. tuberculata is much thicker and corresponds with the coriaceous wall sensu walker (1986). collections examined. poland. all from władysławowo (54º47’n, 18º26’e), in the root zone of melilotus officinalis (l.) pall., 20 sept. 1996, błaszkowski j. 2656-2660 (dpp); under p. lanceolata, 20 sept. 1996, błaszkowski j. 2661-2663 (dpp); from among roots of trifolium repens l., 20 sept. 1996, błaszkowski j. 2664-2669 (dpp). distribution and habitat. in poland, spores of a. rehmii were found in three field-collected samples of roots and rhizosphere soils taken under m. officinalis, p. lanceolata, and t. repens. all the plant species grew in a salt pan near władysławowo at the beginning of the hel peninsula. acaulospora rehmii has originally been isolated from a crop field at caicedonia, valle de cauca, colombia (s i e v e r d i n g , to r o 1987). this fungus has been associated with manihot esculentum, phaseolus, sorghum and crotalaria species. additionally, s i e v e r d i n g and to r o (1987) found spores of a. rehmii in a culture containing a soil sample coming from the centro de pesquisa agropecuária dos cerrados, brasilia d. f., brazil. recently, m o r e i r a -s o u z a et al. (2003) confirmed the presence of this fungus in brazil; a. rehmii spores were encountered among roots of araucaria angustifolia (bert.) o. ktze growing in a reforested area in the state park of campos do jordão, san paulo. wu & l i u (1995) isolated spores of a. rehmii from among roots of phyllostachys pubescens mazel growing in a garden in chi-tou experimental station, national taiwan university. thus, this paper for the first time informs of the occurrence of a. rehmii in europe. mycorrhizal associations. in poland, a. rehmii has been associated with vesicular-arbuscular mycorrhizal roots of p. lanceolata. many attempts to initiate sporulation of this fungus in both trap and one-species cultures failed. according to s i e v e r d i n g & to r o (1987), a. rehmii produced vesicular-arbuscular mycorrhizae. gigaspora margarita w.n. becker et i.r. hall spores produced singly in the soil, blastically at the tip of a bulbous sporogenous cell (fig. 7). spores yellowish white (4a2) to sunflower yellow (4a7); globose to subglobose; (260-)357(-405) μm diam; sometimes ovoid; 300-340 x 360-380 μm (fig. 7). subcellular structure of spores consists of a spore wall comprising two layers (layers 1-3; figs 8 and 9) and a one-layered germination wall. spore wall layer 1, forming the spore surface, permanent, smooth, hyaline, (1.7-)1.9(-2.2) μm thick (figs. 8 and 9), sometimes separating from layer 2 in crushed spores. layer 2 of spore wall laminate, smooth, yellowish white (4a2) to sunflower yellow (4a7), (19.4-)23.4(-26.2) μm acaulospora rehmii and gigaspora margarita 45 thick, consisting of a variable number of laminae, each (1.2-)1.5(-1.7) μm thick, usually separating from each other in crushed spores (figs 8 and 9). germination wall permanent, flexible to semi-flexible, concolorous with spore wall layer 2, (1.5-)1.6(-1.8) μm thick (figs 8 and 9), covered with processes on its under surface (fig. 10); processes 3.7-7.6 x 2.6-3.9 μm, spaced 1.7-13.5 μm from each other; forming prior to germination. in melzer’s reagent, only spore wall layer 2 stains deep red (11c8; fig. 11). sporogenous cell orange (5b8) to brownish yellow (5c8), clavate; 52.5-60.0 x 75.0-100.0 μm (figs 7, 11, 12). wall of sporogenous cell composed of two layers (figs 11 and 12). layer 1 hyaline, (1.0-)1.7(-2.0) μm thick, continuous with spore wall layer 1 (fig. 11). layer 2 orange (5b8) to brownish yellow (5c8), (4.7-)5.6(-6.4) μm thick, continuous with spore wall layer 2 (fig. 11). of the eight described species of the genus gigaspora, b e n t i v e n g a & m o r t o n (1995) accepted five. gigaspora candida bhattacharjee et al., g. ramisporophora spain, sieverd. and n.c. schenck, and g. tuberculata neeraj et al. were considered congeneric with g. rosea nicol. et n.c. schenck, g. margarita, and scutellospora persica (koske et c. walker) c. walker et f.e. sanders, respectively. the most distinctive characters of g. margarita are its yellow-coloured spores (fig. 7), whose colour results from the pigmentation of the spore wall [is not associated with spore contents as in g. gigantea (nicol. et gerd.) gerd. et trappe], and their relatively thin, usually less than 25 μm thick, wall. the only other species of gigaspora forming spores similar in size and colour to those of g. margarita is g. decipiens i.r. hall et l.k. abbott. the character separating the two fungi is the much thicker [(15-)20-45(-90) μm; b e n t i v e n g a, m o r t o n (1995)] spore wall of the former species. all attempts to establish one-species cultures of g. margarita made by the authors of this paper failed. hence, the ontogenetic development and the differentiation of subcellular structures of spores of this fungus were not recognized. according to b e n t i v e n g a & m o r t o n (1995), spores of g. margarita originate blastically from the top of a bulbous sporogenous cell and their wall consists of two, thin layers of near-equal thickness. at first, the spores are white. with time, the inner spore wall layer thickens and the spores darken because of the synthetization of new sublayers from the spore contents. the ontogenetic development of spores ends the formation of a germination wall ornamented with processes prior to germination. the germ tube penetrates through the spore wall. b e n t i v e n g a & m o r t o n (1995) suggested that the two wall layers of spores of g. margarita and all the other gigaspora spp. originating at the same time indicate them to be interdependent elements of one spore wall. a similar spore wall structure occurs in juvenile spores of members of the genus scutellospora. this suggests that gigaspora is ancestral to scutellospora, the only other member of the family gigasporaceae, although molecular analyses of ribosomal dna did not confirm it (s i m o n et al. 1993). compared with fungi of all the other genera of the phylum glomeromycota, species of gigaspora distinguish the lowest morphological variability of their spores. spores of all gigaspora spp. are smooth and the characters separating them are only colour and size of spores, as well as thickness of their wall. except for g. gigantea, whose colour of spores results from the pigmentation of their contents, the spore colour of the other species comes from their wall. b e n t i v e n g a & m o r t o n (1995) 46 j. błaszkowski et al. hypothesized that such low amount of diversity in gigaspora results from two reasons. firstly, germination of gigaspora spp. is associated with spore wall and the energy remained after completion of this vital process probably is too low to aid further differentiation of this wall. in scutellospora spp., germination is taken over by the germination shield, which enables the spore wall to unconstrainedly express variation. secondly, the low level of variability in gigaspora spp. may result from their recent separation and the lack of time to further diverge morphologically. collections examined. poland. szczecin (53º26’n, 14º35’e), under taxus baccata l., 2 may 2001, błaszkowski j. 2670-2672 (dpp); dąbrówka wielkopolska (15o49’e, 52o17’n), from under rumex acetosella l., 22 july 2003, błaszkowski j. 2673 (dpp); chociszewo (15o45’e, 52o19’n), among roots of achillea millefolium l., 22 july 2003, błaszkowski j. 2674 (dpp); and żydowo (15o45’e, 52o24’n), in the rhizosphere of polygonum persicaria l., 3 aug. 2003, błaszkowski j. 2675 (dpp). distribution and habitat. in poland, g. margarita was found in four mixtures of roots and rhizosphere soils, of which three were collected in the lubuskie province and one in the western pomerania province. all the mixtures represented wild plants. of them, only a. millefolium and p. persicaria hosted spores of g. margarita in the field. however, the associations of g. margarita with roots of p. persicaria and t. baccata were revealed only after the cultivation of their field-collected roots and rhizosphere soils in trap cultures. the sporulation of g. margarita in both the field and trap cultures was low. among the 109 spores of arbuscular fungi isolated from 100 g of dry field soil taken under a. millefolium, only one (0.9%) belonged to g. margarita. polygonum persicaria growing in the field hosted a total of 77 spores in 100 g of dry soil, including two (2.6%) of g. margarita. in trap cultures with z. mays as the plant host and mixtures of roots and rhizosphere soils of r. acetosella and t. baccata, the densities of g. margarita spores were 2 and 9 in 100 g of dry soil, respectively, and g. margarita was the only arbuscular fungus revealed. in the field, other species of arbuscular fungi associated with roots of a. millefolium were glomus aggregatum n.c. schenck et s.m. sm. emend. koske, gl. constrictum trappe and scutellospora dipurpurescens j.b. morton et koske, and gl. constrictum and gl. deserticola trappe, bloss et j.a. menge occurred among roots of p. persicaria. gigaspora margarita probably has a worldwide distribution. the holotype of this fungus has been selected from spores recovered from under glycine max (l.) merr. growing in a pot culture containing a root x rhizosphere soil mixture of g. max cultivated at the agronomy south farm of university of illinois, usa (b e c k e r , h a l l 1976). apart from many other reports of the occurrence of gigaspora margarita in both cultivated (agricultural soils, orchards, nurseries) and natural sites (maritime sand dunes, native woodland, prairie) of the us (a n et al. 1993; h e t r i c k , b l o o m 1983; k o s k e , g e m m a 1997; m e n g e et al. 1977; m i l l e r , d o m o t o , wa l k e r 1985; n i c o l s o n , s c h e n c k 1979; r o s e 1988; s c h e n c k , k i n l o c h 1980; s c h e n c k , s m i t h 1981), this species has also been found associated with different cultivated and uncultivated plants of japan (s a i t o , va r g a s 1991), new zealand (h a l l 1977), china (wu et al. 2002), canada (m a r c e l et al. 1988; h a m e l et al. 1994), mexico (e s t r a d a -to r r e s et al. 1992), cuba (f e r r e r , h e r r e r a 1980), and south america (m o r e i r a -s o u z a et al. 2003; s i e v e r d i n g 1989). acaulospora rehmii and gigaspora margarita 47 references a n z. q, h e n d r i x j. w., h e r s h a m n d. e., f e r r i s s r. s., h e n s o n g. t. 1993. the influence of crop rotation and soil fumigation on a mycorrhizal fungal community associated with soybean. mycorrhiza 3: 171 182. b e c k e r w. n., h a l l i. r. 1976. gigaspora margarita, a new species in the endogonaceae. mycotaxon 4: 155 160. b e n t i v e n g a s. p., m o r t o n j. b. 1995. a monograph of the genus gigaspora, incorporating develop mental patterns of morphological characters. mycologia 87: 719 731. b ł a s z k o w s k i j. 1995. glomus corymbiforme, a new species in glomales from poland. mycologia 87: 732 737. b ł a s z k o w s k i j. 2003. arbuscular mycorrhizal fungi (glomeromycota), endogone, and complexipes species deposited in the department of plant pathology, 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223 231. h e t r i c k b. a. d., b l o o m j. 1983. vesicular arbuscular mycorrhizal fungi associated with native tall grass prairie and cultivated winter wheat. can. j. bot. 61: 2140 2146. j a n o s d. p., tr a p p e j. m. 1982. two new acaulospora species from tropical america. mycotaxon 15: 515 522. k o r n e r u p a., wa n s c h e r j. h. 1983. methuen handbook of colour. 3rd ed. e. methuen and co., ltd., london. 252 pp. k o s k e r. e., g e m m a j. n. 1997. mycorrhizae and succession in plantings of beachgrass in sand dunes. am. j. bot. 84: 118 130. m a r c e l g. a., v a n d e r h e i d e n m. g. a., k l i r o n o m o s j. n., u r s i c m., m o u t o g l i s p., s t r e i t w o l f e n g e l r., b o l l e r t., w i e m k e n a., s a n d e r s i. r. 1988. mycorrhizal fungal diversity determines plant biodiversity, ecosystem variability and productivity. nature 396: 69 72. m e n g e j. a., n e m e c s., d a v i s r. m., m i n a s s i a n v. 1977. mycorrhizal fungi associated with citrus and their possible interactions with pathogens. proc. int. soc. citriculture 3: 872 876. m i l l e r d. d., d o m o t o p., wa l k e r c. 1985. mycorrhizal fungi at eighteen apple rootstock plantings in the united states. new phytol. 100: 379 391. m i r e k z., p i ę k o ś m i r k o w a h., z a j ą c a., z a j ą c m. 1995. vascular plants of poland. a checklist. polish botanical studies, guidebook 15, kraków. 303 pp. m o r e i r a s o u z a m., tr u e m s. f. b., g o m e s d a c o s t a s. m. c a r d o s o e. j. b. n. 2003. ar buscular mycorrhizal fungi associated with araucaria angustifolia (bert.) o. ktze. mycorrhiza 13: 211 215. m o r t o n j. b. 1986. three new species of acaulospora (endogonaceae) from high aluminum, low ph soils in west virginia. mycologia 78: 641 648. m o r t o n j. b. 2002. international culture collection of vesicular) arbuscular mycorrhizal fungi. west virginia university: http://www.invam.caf.wvu.edu/. n i c o l s o n t. h., s c h e n c k n. c. 1979. endogonaceous mycorrhizal endophytes in florida. mycologia 71: 178 198. o m a r m. b., b o l l a n l., h e a t h e r w. a. 1979. a permanent mounting medium for fungi. bull. brit. mycol. soc. 13: 31 32. p h i l l i p s j. m., h a y m a n d. s. 1970. improved procedures for clearing roots and staining parasitic and vesicular arbuscular mycorrhizal fungi for rapid assessment of infection trans. brit. mycol. soc. 55: 158 161. 48 j. błaszkowski et al. r o s e s. 1988. above and belowground community development in a maritime sand dune ecosystem. plant and soil 109: 215 226. r o t h w e l l f. m., tr a p p e j. m. 1979. acaulospora bireticulata sp. nov. mycotaxon 8: 471 475. s a i t o m., va r g a s r. 1991. vesicular arbuscular mycorrhizal fungi in some humus rich ando soils of japan. soil microorg. 38: 3 15. s c h e n c k n. c., k i n l o c h r. a. 1980. incidence of mycorrhizal fungi on six field crops in monoculture on a newly cleared woodland site. mycologia 72: 445 456. s c h e n c k n. c., s m i t h g. s. 1981. distribution and occurrence of vesicular arbuscular mycorrhizal fungi on florida agricultural crops. soil and crop sci. soc. florida 40: 171 175. s i e v e r d i n g e. 1988. two new species of vesicular arbuscular mycorrhizal fungi in the endogonaceae from tropical high lands of africa. angew. bot. 62: 373 380. s i e v e r d i n g e. 1989. ecology of vam fungi in tropical ecosystems. agric., ecosyst. and environ. 29: 369 390. s i e v e r d i n g e., to r o s. t. 1987. acaulospora dendiculata sp. nov. and acaulospora rehmii sp. nov. (endogonaceae) with ornamented spore walls. angew. bot. 61: 217 223. s i m o n l., b o u s q u e t t., l é v e s q u e r. c., l a l o n d e m. 1993. origin and diversification of endomy corrhizal fungi and coincidence with vascular land plants. nature 363: 67 69. s t ü r m e r s. l., m o r t o n j. b. 1997. developmental patterns defining morphological characters in spores of four species in glomus. mycologia 89: 72 81. wa l k e r c. 1983. taxonomic concepts in the endogonaceae: spore wall characteristics in species de scriptions. mycotaxon 18: 443 455. wa l k e r c. 1986. taxonomic concepts in the endogonaceae. ii. a fifth morphological wall type in en dogonaceous spores. mycotaxon 25: 95 99. wa l k e r c., tr a p p e j. m. 1981. acaulospora spinosa sp. nov. with a key to the species of acaulospora. mycotaxon 12: 515 521. wa l k e r c., tr a p p e j. m. 1993. names and epithets in the glomales and endogonales. mycol. res. 97: 339 344. w u t., h a o w., l i n x., s h i y. 2002. screening of arbuscular mycorrhizal fungi for the revegetation of eroded red soils in subtropical china. plant and soil 239: 225 235. w u c. g., l i u y. s., h w u a n g y. l., wa n g y. p., c h a o c. c. 1995. glomales of taiwan: v. glomus chimonobambusae and entrophospora kentinensis, spp. nov. mycotaxon 53: 283 294. acaulospora rehmii i gigaspora margarita, arbuskularne grzyby mikoryzowe (glomeromycota) nowe odpowiednio dla europy i polski s t r e s z c z e n i e opisano i zilustrowano cechy morfologiczne zarodników acaulospora rehmii i gigaspora margarita, arbuskularnych grzybów mikoryzowych z gromady glomeromycota. zarodniki obu tych gatunków znaleziono w polowych mieszaninach gleby ryzosferowej i korzeni zebranych w polsce. próby otrzymania zarodników tych grzybów w kulturach pułapkowych powiodły się tylko z g. margarita. wszystkie próby ustanowienia jednogatunkowych kultur obu grzybów nie udały się. gigaspora margarita znaleziono po raz pierwszy w polsce, a a. rehmii po raz pierw szy w europie. przedstawiono również poznane rozmieszczenie obu gatunków w świecie. 2014-01-01t11:43:22+0100 polish botanical society book review besl h., bresinsky a. 2009. checkliste der basidiomycota von bayern (agaricomycotina, urediniomycotina, ustilaginomycotina). regensburger mykologische schriften, band 16, regensburg, 868 pp. issn 0944-2820. the checklist is published as a part of the series renowned for presenting of the biodiversity of plants and fungi in germany. the current volume offers an extended overview of the history of former studies as well as the current state of knowledge of basidiomycetous fungi in bavaria. material is grouped in three main units, with the genera and species arranged alphabetically within each unit. first part of the checklist consists of phytopathogenic microfungi classified in urediniomycotina and ustilagomycotina. these taxa are merged together into one list as they share basic ecological properties whereas the species belonging to agaricomycotina are split into two groups depending on the morphological structure of their fruitbodies. the first group covers agaricoid, boletoid and cyphelloid taxa while the second one deals with aphyllophoroid, heterobasidioid and gastroid species. for each of the species, the following information is provided: correct name, synonyms, region of occurrence (defined according to the map given in the introductory chapter and often supplemented with an altitude value), some ecological data (substrate or name of the species hosting parasite) and stage of the fungus life cycle (in the case of rust fungi). the occurrence of the species is based on the references to the sources reporting it as well as on the exsiccata studied. they were facilitated mostly by fungal collections of the botanische staatssammlung münchen (m) and of the herbarium of the university in regensburg (reg), although other herbaria (including private ones) contributed to the checklist as well. such professionally preparing of the checklist provides an opportunity for revision and confirmation of many literature data and guarantees reliable effect. the species of special interest are annotated with taxonomical comments and endangered taxa are given the status according to the red list criteria. the book offers far more than just a checklist of fungal taxa. it is a valuable contribution to the knowledge of chorology and ecology of many basidiomycetous fungi occurring in a region rich of diversified habitats. m. ruszkiewicz-michalska acta mycologica vol. 45 (1): 125 2010 2014-01-01t11:50:50+0100 polish botanical society yeast-like fungi isolated in students anna biedunkiewicz department of mycology, university of warmia and mazury in olsztyn oczapowskiego 1a, pl-10-957 olsztyn, alibi@uwm.edu.pl b i e d u n k i e w i c z a.: yeast-like fungi isolated in students. acta mycol. 42 (1):141-149, 2007. the occurrence of yeast-like fungi in the most important infection portals of the respiratory system in 200 randomly chosen students of biology and veterinary medicine was examined. the students come into direct contact with plants and animals that may be colonised by fungi belonging to various systematic groups. nine species of yeast-like fungi, including 7 species determined in the biologists, were recorded in the subjects. candida tropicalis and c. albicans were the most frequently isolated fungi. the greatest number of fungi was isolated from the oral cavity (124 isolates), fewer from the throat (79 isolates), and the smallest number from the nose (8 isolates). fungi occurred more frequently in autumn and slightly less frequently in spring, and were isolated more frequently from women than from men. key words: yeast-like fungi, students, respiratory system, infection introduction a continuous increase in fungal infections has been recorded in recent years (b a r a n 1998). it is believed that they affect ca. 40% of the world population (m a d d i n 1992). however, studies on the occurrence of yeast-like fungi mostly concentrate on compromised or hospitalised individuals under 20 or over 40 years of age (b i e d u n k i e w i c z 1999, 2001; d y n o w s k a 1990, 1993, 1995) while yeastlike fungi can also occur as commensals in various ontocoenoses in healthy individuals (b a t u r a g a b r y e l , f i r l i k , w i e c z o r e k 1994; b a r a n 1998; r i c h a r d s o n , wa r n o c k 1995). they colonise the oral cavity, mucous membranes, the skin, digestive system or genitourinary system. plants and animals can also be reservoirs of fungi (b i e g a ń s k a , d w o r e c k a k a s z a k 2004; d y n o w s k a 1995; k u r n a t o w s k a 1999). therefore mycological studies were conducted on a group of healthy students of biology and veterinary medicine who come into direct contact with plants or animals, both living and dead, that may be colonised by fungi. acta mycologica vol. 42 (1): 141-149 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday yeast-like fungi 145 ta b l e 2 growth intensity of yeast-like fungi in autumn and spring study season/ faculty growth intensity number of ontocoenoses autumn b autumn vm spring b spring vm autumn sum spring sum poor 31 22 32 28 53 60 moderate 10 12 6 6 22 12 abundant 5 7 4 8 12 12 very abundant 3 7 2 6 10 8 sum 49 48 44 48 97 92 explanations: autumn b number of isolates obtained from the students of biology in autumn; autumn vm number of isolates obtained from the students of veterinary medicine in autumn ta b l e 3 number of positive cultures in men and women in autumn and spring study season/ faculty sex number of positive cultures autumn spring autumn spring b % vm % b % vm % sum % sum % women 36 73,47 27 71,05 27 61,36 25 65,79 63 73,26 52 63,41 men 13 26,91 11 28,95 17 38,64 13 34,21 23 26,74 30 36,59 sum 49 100 38 100 44 100 38 100 86 100 82 100 ta b l e 4 occurrence of fungi in coffee drinkers, alcohol drinkers and smokers type of substance positive cultures (%) biology veterinary medicine alcohol 12,0 25,0 coffee 30,0 51,7 cigarettes 19,5 63,6 ta b l e 5 occurrence frequency of diseases preceding sampling disease rarely often very often total throat diseases 21 28 13 62 sinus diseases 13 9 5 27 bronchial diseases 22 5 2 29 pneumonia 10 1 11 rhinitis 29 42 20 91 ear infection 11 4 1 16 asthma 1 2 3 allergies 5 6 3 14 tuberculosis diabetes vitamin deficiencies 3 3 1 7 internal mycoses. 5 4 9 cutaneous mycoses 7 1 8 thyroid diseases renal diseases 8 1 9 146 a. biedunkiewicz results was shown only among the subjects preferring animal foodstuffs (among the biology students: 79%) and the subjects preferring dairy products (among the veterinary students: 46.9%). diets rich in carbohydrates were not reported. questionnaire data also show that infections of the throat, bronchii and sinuses, and catarrhal conditions were the most frequent disorders preceding the examination (tab. 5). discussion yeast-like fungi that commonly occur in human environments colonise various ontocoenoses relatively easily. they enter the human body via the oral or nasal cavities when ingested or inhaled. the increase in fungal infections recorded worldwide is caused by the growing number of predisposing factors and the simultaneous decrease in natural immunity of healthy individuals (g h a n n o u m , a b u e l t e e n 1990; s z y m a n i a k et al. 2005). as a result, permeability of the epithelium changes, allowing fungi to enter the body and penetrate the tissues (d y n o w s k a , b i e d u n k i e w i c z , s u c h a r z e w s k a 2002). fungi isolated in such portals of infection as the oral cavity can be causal agents of the development of fungal infections although they do not have to. k u r n a t o w s k i and r a c z y ń s k a w i t o ń s k a (2004) recorded fungi in the oral cavity of 65.6% of the individuals studied. they detected: candida albicans (43.8%), c. tropicalis (15.7%), c. krusei (9.7%), c. kefyr (9.7%), c. guilliermondii (4.9%), c. parapsilosis (4.3%), c. glabrata (5.9%) and geotrichum candidum (5.9). yeast-like fungi, mostly candida albicans, are recorded asymptomatically in the oral cavity in 40-60% of individuals (k a ł o w s k i 1964). d a r w a z e h , a l r e f a i and a l m o j a i w e l (2001) believe that fungi of the genus candida are part of the normal, commensal mycoflora of the oral cavity, present in 30-70% of generally healthy individuals. in mass studies, k o w s z y k g i n d i f e r and s o b i c z e w s k i (1986) recorded fungal infections in the oral cavity in 44% of students and 46% of individuals across various occupational groups. in the studies conducted by k u r n a t o w s k a (2003) on individuals suffering from conditions of the oral cavity, as many as 6 species of the genus candida were isolated: c. albicans – 146 strains (74.6%), c. tropicalis – 18 (9.2%), c. krusei – 10 (5.1%), c. guilliermondii – 6 (3.1%), c. famata – 5 (2.6%), c. zeylanoides – 2 (1.0%). candida albicans was also the most frequently recorded fungus in studies by m a j e w s k a et al (2000) who additionally observed a high frequency of c. glabrata, c. parapsilosis and c. tropicalis. the differences in the frequency of occurrence of these fungi may be related to the season, time of day, type of diet, oral hygiene, as well as sampling and test methods. candida tropicalis was one of the most frequently isolated species of yeast-like fungi: 52% among the biology students and 31% among the veterinary students. this finding is corroborated in studies by e j d y s and d y n o w s k a (2004) conducted on a group of healthy children. the majority of species recorded in the present study are classified as biosafety level 2 (bsl2): they may cause severe, opportunistic infections in immunocompromised individuals (d e h o o g 1996). the occurrence of fungi in the nasal cavity or paranasal sinuses in healthy individuals is reported very rarely. their presence in these ontocoenoses is mostly related to chronic inflammatory conditions of mucous membranes caused by bacteria, yeast-like fungi 147 allergy, asthma, polyps or deviated nasal septum (h o f m a n , s k r o b i s z and h o f m a n 2004; j a n k o w s k a k o n s u r , p r e ś and b a r a n 2005). k o w s z y k g i n d i f e r and s o b i c z e w s k i (1986) recorded fungi in 69% of the students examined by them. k u r n a t o w s k i and r a c z y ń s k a -w i t o ń s k a (2004) report that fungi occur on the mucous membrane of the throat in 24.5-53.6% of patients suffering from chronic pharyngitis, in ca. 2/3 of conditions of palatine tonsils and only in 1/5 of those of the pharyngeal tonsil. fungi are recorded in one or a few ontocoenoses in the same person. the present results correspond to those obtained by e j d y s and d y n o w s k a (2004), who observed this correlation in a group of healthy children in their study on environmental preconditioning of fungal infections. the occurrence of potentially pathogenic fungi in individuals who consider themselves to be generally healthy may be related to a decrease in their immunity. sources of fungal infection should thus be considered. as reported in the literature, generalised or multifocal infections caused by fungi of the genus candida are mostly endogenous, which does not exclude their exogenous origin (b i e g a ń s k a , d w o r e c k a k a s z a k 2003). animals can also be a source of infection. students of biology and veterinary medicine come into direct contact with animals, both healthy and ill, during classes. some, especially veterinary students, may work in veterinary clinics where they deal with animals that are living or dead (in anatomy laboratories). the occurrence of fungi of the genus candida is recorded in many animal species and diseases caused by these fungi are not rare. the literature published so far does not show whether interspecific transmission of candida infection is possible and what the particular character of biotypes of individual species of these yeast-like fungi in relation to the host is. b i e g a ń s k a and d w o r e c k a k a s z a k (2003) examined 230 candida strains isolated from animals between 1998 and 2002. they noticed an increase in the number of infections caused by these fungi in animals, and the number of all positive results exceeded 15% in 2002. this means that candida sp. is an important aetiological factor of infection, also in animals. candidiases were detected mostly in dogs, horses and birds, and affected the respiratory system (nose and throat), reproductive system, skin and its products (b i e g a ń s k a , d w o r e c k a k a s z a k 2003). as can be seen, no great differences exist in specific places of isolation of candida sp. between people and animals. zoonoses may in many cases be treated as occupational diseases. particularly exposed occupational groups include veterinarians, veterinary technicians, students of these subjects, technicians specialising in animal husbandry, animal farmers, staff in vivaria, zoological gardens, abattoirs and utilisation plants, etc. (d u t k i e w i c z , j a b ł o ń s k i 1989; f i j a ł k o w s k a 1983). isolating potentially pathogenic fungi in healthy individuals indicates decreasing immunity. commensal fungi can become causal agents of a number of diseases. this shows the need for a greater emphasis on mycological tests and analyses, also in outpatients, to increase immunoprevention or, directly, antifungal prevention (m o c h o n , c u t l e r 2005). 148 a. biedunkiewicz conclusions 1. nine species of yeast-like fungi belonging to 3 genera: candida, saccharomyces and saccharomycopsis, were recorded in healthy students. candida tropicalis (83 isolates) and candida albicans (77 isolates) were the most frequently isolated fungi. 2. the greatest number of fungi were isolated from the oral cavity (124 isolates), fewer from the throat (79 isolates), and the smallest number from the nose (8 isolates). 3. the greatest percentage of positive results was recorded in the subjects reporting excessive coffee consumption among the biology students (30%), and cigarette smoking (63.6%) and excessive alcohol consumption (51.7%) among the veterinary students. 4. isolating potentially pathogenic fungi in healthy individuals in the 20-30 age bracket indicates their decreasing immunity. it shows the need for a greater emphasis on mycological testing to increase antifungal prevention and help identify carriers of fungi among students. references b a r a n e. 1998. zarys mikologii lekarskiej. volumed. wrocław. b a r n e t t j. a., p a y n e r. w., ya r r o w d. 1990. yeasts: characteristics and identification. cambridge univ. press. b a t u r a -g a b r y e l h., f i r l i k m., w i e c z o r e k u. 1994. ocena występowania zakażenia grzybiczego u chorych z rakiem płuc. med. dośw. mikrobiol. 46: 77–81. b i e d u n k i e w i c z a. 1999. analiza mikologiczna materiału bronchoskopowego. mikol. lek. 6 (1): 33–40. b i e d u n k i e w i c z a. 2001. dynamics of human respiratory system mycoflora. acta mycol. 36 (2): 211– 239. b i e d u n k i e w i c z -z i o m e k a., d y n o w s k a m. 2004. candida dubliniensis sullivan et al., a new species in the human respiratory system. acta mycol. 39 (1): 7–12. b i e g a ń s k a m., d w o r e c k a -k a s z a k b. 2003. charakterystyka szczepów candida sp. izolowanych w latach 2001-2002 od zwierząt. mikol. lek.10 (4): 275–283. d a r w a z e h a. m., a l -r e f a i s., a l-m o j a i w e l s. 2001. isolation of candida species from the oral cavity and fingertips of complete denture wearers. the journal of prosthetic dentistry 86 (4): 420– 423. de h o o g g. s. 1996. risk assessment of fungi reported from humans and animals. mycoses 39: 407– 417. d e h o o g g. s., g u a r r o j., f i g u e r r a s m. j. 2000. atlas of clinical fungi. utrecht, the netherlands, reus, spain. d u t k i e w i c z j., j a b ł o ń s k i j. 1989. biologiczne szkodliwości zawodowe. pzwl. warszawa. d y n o w s k a m. 1990. występowanie grzybów z rodzaju candida w układzie oddechowym mieszkańców województwa olsztyńskiego. acta mycol. 26 (1): 99–107. d y n o w s k a m. 1993. observations concerning the appearance of yeasts in human respiratory system. acta mycol. 28 (2): 147–150. d y n o w s k a m. 1995. drożdże i grzyby drożdżopodobne jako czynniki patogenne i bioindykatory ekosystemów wodnych. studia i materiały wsp 77. olsztyn. d y n o w s k a m., b i e d u n k i e w i c z a. 1999. presence of saccharomycopsis capsularis in the human respiratory system. acta mycol. 34 (2): 281–287. d y n o w s k a m., biedunkiewicz a., sucharzewska e. 2002. participation of yeast-like fungi in respiratory system diseases, tuberculosis and neoplasms. acta mycol. 37 (1/2): 117–122. e j d y s e., d y n o w s k a m. 2004. environmental preconditioning of the fungal infection of children. mikol. lek. 11 (1): 9-13. f i j a ł k o w s k a w. 1983. czym mogą zarazić nas zwierzęta? pwril. warszawa. g h a n n o u m m. a., a b u -e l t e e n k. m. 1990. pathogenicity determinants of candida. mycoses 33: 265–282. yeast-like fungi 149 h o f m a n t., s k r o b i s z w., h o f m a n a. 2004. ocena skuteczności leczenia flukonazolem chorych na przewlekłe zapalenie zatok przynosowych. mikol. lek. 11 (4): 297–301. h o w a r d d. h. 2003. pathogenic fungi in humans and animals. marcel dekker inc., new york, basel. j a n k o w s k a -k o n s u r a., p r e ś k., b a r a n e. 2005. grzybicze zakażenia nosa i zatok przynosowych. mikol. lek. 12 (2): 133:136. k a ł o w s k i m. 1964. występowanie drożdżaków w zdrowej jamie ustnej człowieka. czas. stomatol. 17: 427–432. k o w s z y k -g i n d i f e r z., s o b i c z e w s k i w. 1986. grzybice i sposoby ich zwalczania. pzwl. k r e g e r -va n r i j n. j. w. 1984. the yeasts: a taxonomy study. third revision and enlarged edition. els. sci. publ. amsterdam. k u r n a t o w s k a a. 1995. wybrane zagadnienia z mikologii medycznej. promedi. łódź. k u r n a t o w s k a a. 1999. ekologia. jej związki z różnymi dziedzinami wiedzy. pwn. warszawa. k u r n a t o w s k a a. j. 2003. występowanie grzybów w ontocenozie jamy ustnej a zmiany błony śluzowej. mikol. lek. 10 (4): 295–298. k u r n a t o w s k i p., r a c z y ń s k a -w i t o ń s k a g. 2004. grzyby jako czynnik etiologiczny chorób górnych dróg oddechowych. wiad. parazytol., 50 (2): 157–162. m a c u r a a. b. 1987. przyleganie grzybów drożdżopodobnych do komórek ssaków. post. mikrobiol. 26: 337–351. m a d d i n s. 1992. what is the therapeutics? j. europ. acad. dermatol. venerol. 1: 31–35. m a j e w s k a a., s o z a ń s k a z., k a s i a k m., c z a j c z y ń s k a -wa s z k i e w i c z a. 2000. występowanie grzybów drożdżopodobnych w jamie ustnej a intensywność próchnicy zębów. mikol. lek. 7 (2): 71–75. m o c h o n a. b., c u t l e r j. e. 2005. prospects of vaccines for medically important fungi. medical mycol-prospects of vaccines for medically important fungi. medical mycol-medical mycology 43: 97–115. r a b c z y ń s k i j., d z i ę g i e l p., z i ó ł k o w s k i p. 1998. uogólniona drożdżyca u noworodków – obraz patomorfologiczny. mikol. lek. 5 (2): 105–109. r i c h a r d s o n m. d., wa r n o c k d. w. 1995. grzybice – rozpoznawanie i leczenie. pwn. warszawa. s a m o n i s g. 2004. gut: portal of entry of fungi in the immunocompromised hosts. mikol. lek.11 (2): 105–107. s y s ł o j., m a c u r a a. b. 1998. badania nad niektórymi determinantami patogenności u grzybów z rodj., m a c u r a a. b. 1998. badania nad niektórymi determinantami patogenności u grzybów z rod-m a c u r a a. b. 1998. badania nad niektórymi determinantami patogenności u grzybów z roda. b. 1998. badania nad niektórymi determinantami patogenności u grzybów z rodzaju candida. mikol. lek. 5 (3): 149–155. s z y m a n i a k l., w o j c i e c h o w s k a -k o s z k o i., k l i m o w i c z b., g i e d r y s -k a l e m b a s. 2005. non-lipophilic yeast flora from selected body sites in healthy subjects. mikol. lek. 12 (4): 291–295. grzyby drożdżopodobne izolowane od studentów s t r e s z c z e n i e w pracy przeanalizowano występowanie grzybów drożdżopodobnych w głównych wrotach zakażenia układu oddechowego, dwustu losowo wybranych studentów biologii i medycyny weterynaryjnej, mających w trakcie studiów bezpośredni kontakt z roślinami i zwierzętami, które mogą być zasiedlane przez grzyby z różnych grup systematycznych. w przeprowadzonych badaniach u wszystkich studentów stwierdzono 9 gatunków grzybów drożdżopodobnych, z czego 7 u biologów. najczęściej izolowano: candida tropicalis (83 izolaty) i candida albicans (77 izolatów). najwięcej grzybów wyizolowano z jamy ustnej (124 izolaty), mniej z gardła (79), najmniej z nosa (8). grzyby częściej występowały jesienią, nieco rzadziej wiosną, u kobiet niż u mężczyzn. ogółem najwyższy odsetek wyników dodatnich odnotowano u osób zgłaszających nadużywanie kawy (30%), palących papierosy (63,6%) i nadużywających alkoholu (51,7%). izolowanie grzybów potencjalnie chorobotwórczych od zdrowych studentów wskazuje na obniżony stan ich odporności i skłania do położenia większego nacisku na analizy i badania mikologiczne w celu zwiększenia profilaktyki przeciwgrzybicznej w grupach osób zdrowych. 2014-01-01t11:45:49+0100 polish botanical society autofluorescence of ascospores and conidial spores in selected lichen species of the opegrapha genus anetta wieczorek department of ecology, university of szczecin wąska 13, pl-71-415 szczecin, anettaw@univ.szczecin.pl wieczorek a.: autofluorescence of ascospores and conidial spores in selected lichen species of the opegrapha genus. acta mycol. 43 (1): 99–103, 2008. the paper presents findings of the research work on autofluorescence phenomenon in ascospores and conidial spores that was carried out on 12 selected species of the opegrapha genus occurring in poland. key words: lichens, opegrapha, autofluorescence introduction autofluorescence is one of luminescence types and consists in emitting the own light, which is induced by photon capture by a body. own luminescence of cells depends on the presence of endogenous chromophores in tissue. the phenomenon of autofluorescence is frequent not only in plants but also in fungi (rost 1992). it has been observed, among others, in the cytoplasm of ascospore and konidial spore hyphae (žižka, gabriel 2006). studies on the usefulness of autofluorescence methods in identification of fungi species are used in examination of mycorrhiza phenomenon (séjalon-delmas et al. 1998) and in laboratory diagnostics, in particular of pathogenic fungi, and consist among other in determination of tissue colour differences in healthy plants and animals and those pathologically changed (margo, bombardier 1985; elston 2001). in lichens, autofluorescence phenomenon has been used, among others, in evaluation of the viability of hyphae and algal cells according to the method of their storage (honegger 2003) as well as in evaluation of damages in the chlorophyll apparatus of algal cells induced by high pollutant concentration (favali et al. 1991; silberstein et al. 1996). sometimes, this phenomenon can be used for determining germination capacity (cathy, warren 1984). this study aimed at taking up an attempt to use the autofluorescence phenomenon in identification of lichen species of the opegrapha genus. acta mycologica vol. 43 (1): 99–103 2008 100 a. wieczorek material and methods in the study were used selected epiphytic and epilitic species of the opegrapha genus occurring in poland (o. atra pers., o. calcarea sm., o. culmigena lib., o. dolomitica (arnold) clauzade & cl. roux, o. gyrocarpa flot., o. niveoatra (borrer) j. r. laundon, o. rufescens pers., o. rupestris pers., o. varia pers., o. vermicellifera (kunze) j. r. laundon, o. viridis (ach.) behlen & desberger, and o. vulgata (ach.) ach. species terminology was adopted after santesson et al. (2004) and dietrich et al. (2008). in order to determine the intensity of autofluorescence, a series of microscope slides was prepared, 10 samples per each species, using the fresh material collected by the author from the bark of different tree species within the area of the western poland in 2004-2006 as well as the herbarium material. the samples collected from the herbarium material were from 1940-2001. in order to examine in detail the subtle structures, such as ascospores and conidial spores, squashed microscope slides were prepared. in the study, a lsm 510 laser scanning microscope (zeiss), operating on an axiovert 200m motorised inverted microscope with fluorescence (zeiss), was used. the used laser was an argon-ion laser with 11% power output, emitting 488 nm light waves, equipped with a 63x objective for ascospores and a 100x objective for conidial spores and two red filters (between 505 nm–650 nm wavelength) and one green filter (to 505 nm wavelength); when observing ascospores and conidial spores, a nomarski filter was used. results and discussion in all examined species, the emission of light within the wavelength range from 488 nm to 505 nm, and sometimes over 650 nm, was found (tab. 1). the examined taxa were separated into three groups with different fluorescence intensity. the ascospores of the following species were in the first group: o. atra (fig. 1), o. calcarea, o. dolomitica (fig. 2) and o. rupestris (fig. 3), with the fluorescence intensity within both wavelength ranges being clearly observable. in the second group, the spores of o. gyrocarpa, o. vermicellifera, o. vulgata (fig. 4), o. viridis and o. culmigena were found, the autofluorescence intensity of which was much weaker or almost completely inconspicuous. in case of the species of the second group, no expected effects were obtained despite intensification of autofluorescence with ultraviolet radiation. on the other hand, a more rapid burning of autofluorescence was observed after application of ultraviolet radiation. into the third group, the ascospores of such species as o. niveoatra, o. varia (fig. 5) and o. rufescens (fig. 6) were included, in which a variable intensity of fluorescence was found. different level of autofluorescence intensity in these species can be induced on the one hand by a different depth of spore location in microscope slide but on the other hand by a different degree of ascospore maturity. it should be emphasised that observation of spores at the stage of transversal wall formation (figs 7-9) in the taxa with weak autofluorescence showed the occurrence of such phenomenon. in the species with intense autofluorescence, a fading of red light emission was observed as deeper spore layers were scanned due to autofluorescence burning. autofluorescence of ascospores 101 no fundamental differences were observed in the intensity of spore fluorescence in microscope slides prepared both from the fresh and herbarium materials. the conidial spores of all examined species showed similar autofluorescence, irrespective of the origin of the analysed material and their size (figs 10-12). the taken up research is the first attempt of using autofluorescence phenomenon for identification of lichen species of the opegrapha genus. the carried out analyses showed the presence of endogenous chromophores in the cytoplasm of ascospres and conidial spores in all examined taxa. in the literature a correlation was showed between the fluorescence intensity and the viability, maturity degree and storage conditions of ascospores and conidial spores (cathy, warren 1984; honegger 2003). it was also proved that the fluorescence intensity of algal cells changed depending on the degree of environmental pollution (di toppi et al. 2005) and other factors. in the work by favali et al. 1991, not only changes in autofluorescence intensity connected with the harmful effect of such chemical elements as aluminium (al), silicon (si), phosphorus (p), sulphur (s), chlorine (cl), and calcium (ca) were demonstrated but also differences were showed in the concentration of respective chemical elements in different elements of the thallus, including a considerable concentration of aluminium in the apothecia. it is not excluded that these stress factors could also affect the intensity of light emission in ascospores and conidial spores. autofluorescence depends also on the type and the amount of chromophore occurring in the cytoplasm. the carried out analyses of images show a variable autofluorescent response of ascospores in o. niveoatra, o. rufescens and o. varia. the spectrum of luminescence in the examined taxa may undergo changes and is not stable. thus, it is difficult to determine simple criteria that would describe a given group of species, while autofluorescence intensity can not be a trait of diagnostic importance since it may be affected by many external factors. it appears thus that it is not a good method for identification of taxa, at least in that group of species. nevertheless, it is worth expanding the research work on the usefulness of that method in taxonomic studies of other lichen and fungi genera. ta b l e 1 autofluorescence intensity of ascospores in selected lichen species of the opegrapha genus species light emission intensity within wavelength range 488-659nm 488-505 nm high weak very weak o. atra pers. + o. calcarea sm. + o. dolomitica (arnold) clauzade & cl. roux + o. rupestris pers. + o. varia pers. + + o. rufescens pers. + + + o. niveoatra (borrer) j. r. laundon + + o. gyrocarpa flot. + o. vermicellifera (kunze) j. r. laundon + o. vulgata (ach.) ach. + o. viridis (ach.) behlen & desberger + o. culmigena lib. + 102 a. wieczorek conclusions no fundamental taxonomic differences were found at the level of light emission intensity. the autofluorescent response of particular species results most probably from the size of spores and the same from the amount of cytoplasm inside them as well as from the degree of their maturity. as is showed by examinations, dead spores devoid of the cytoplasm as well as those being dormant do not show autofluorescence (cathy, warren 1984). the intensity of fluorescence in ascospores and conidial spores in the genus opegrapha does not seem to be a good method in taxonomic identification of particular species. acknowledgements. i would like to thank aleksander ratajczak, msc, from the laboratory of electron and confocal microscopy of the adam mickiewicz university in poznań for his assistance and kindness with carrying out microscope observations as well as prof. krystyna czyżewska (university of łódź) for her valuable suggestions and comments during preparation of this paper. references cathy h. wu., warren h. l. 1984. natural autofluorescence, and its correlation with viability. mycologia 76 (6): 1049–1058. diederich p., ertz d., stapper n., sérusiaux e., ries c. 2008. the lichens and lichenicolous fungi of belgium, luxembourg and northern france. url: http://www.lichenology.info [april 2008]. di toppi l.s., musetti r., vattuone z., pawlik-skowrońska b., fossati f., bertoli l., badiani m., favali m. a. 2005. cadmium distribution and effects on ultrastructureand chlorophyll status in photobionts and mycobionts of xanthoria parietina. microscopy research and technique 66 (5): 229–238. elston d.m. 2001. fluorescence of fungi in superficial and deep fungal infections. bmc microbiology 1: 21. favali m.a., corradi m.g., fossati f. 1991. x-ray microanalysis and ultrastructure of lichens from polluted and unpolluted areas. plants for toxicity assessment 2: 276–286. honegger r. 2003. the impact of different long-term storage conditions on the viability of lichen-forming ascomycetes and their green algal photobiont, trebouxia spp. plant biology 5 (3): 324–330. margo c. e., bombardier t. 1985. the diagnostic value of fungal autofl uorescence. survey of ophthal-the diagnostic value of fungal autofluorescence. survey of ophthalmology 29: 374–376. rost f.w.d. 1992. fluorescence microscop. cambridge university press, cambidge, 267 pp. santesson r., moberg r., nordin a., tønsberg t., vitikainen o. 2004. lichen-forming and lichenicolous fungi of fennoscandia. museum of evolution, uppsala university, 359 pp. séjalon-delmas n., magnier a., douds d.d., bécard g. 1998. cytoplasmic autofluorescence of an arbuscular mycorrhizal fungus gigaspora gigantea and nondestructive fungal observations in planta. mycologia 90 (5): 921–926. silberstein l., siegel b.z., siegel s.m., mukhtar a., galun m. 1996. comparative studies on xanthoria parietina, a pollution resistant lichen, and ramalina duriaei, a sensitive species. i. effects of air pollution on physiological processes. the lichenologist 28: 355–365. žižka z., gabriel j. 2006. primary fluorescence (autofluorescence) of fruiting bodies of the wood-rotting fungus fomes fomentarius. folia microbiologica 51 (2): 109–113. autofluorescence of ascospores 103 autofluorescencja zarodników workowych i konidialnych wybranych gatunków porostów z rodzaju opegrapha s t r e s z c z e n i e w pracy przedstawiono wyniki badań nad zjawiskiem autofluorescencji zarodników workowych i konidialnych wybranych gatunków porostów z rodzaju opegrapha. zaobserwowano i scharakteryzowano zjawisko autofluorescencji u 12 gatunków występujących w polsce (o. atra, o. calcarea, o. culmigena, o. dolomitica, o. gyrocarpa, o. niveoatra, o. rufescens, o. rupestris, o. varia, o. vermicellifera, o. viridis, o. vulgata). w badaniach wykorzystano laserowy mikroskop skaningowy lsm 510 (zaiss) pracujący na odwróconym mikroskopie axiovert 200m (zaiss) z fluorescencją. zastosowano laser argonowy o mocy 11% wysyłający fale o długości 488 nm, obiektyw 63x dla zarodników workowych i 100x dla konidialnych oraz dwa filtry, czerwony o długości fal od 505 do 650 nm i zielony o długości, do 505 nm. przy obserwacji zarodników wykorzystano filtr nomarskiego. nie stwierdzono zasadniczych różnic taksonomicznych w poziomie intensywności emisji światła. zaobserwowane różnice w intensywności świecenia poszczególnych gatunków wynikają prawdopodobnie z wielkości, a tym samym ilości cytoplazmy wewnątrz zarodników oraz stopnia ich dojrzałości. po raz pierwszy opisano wykorzystanie zjawiska autofluorescencji w badaniach taksonomicznych porostów z rodzaju opegrapha. zaprezentowano pierwsze zdjęcia tego zjawiska, które uzupełniają oryginalny opis rodzaju. 2014-01-01t11:47:32+0100 polish botanical society influence of osmotic pressure on the growth of three species of genus zoophthora jerzy piątkowski and ewa kliczkowska institute of genetics and microbiology, university of wrocław przybyszewskiego 63/77, pl-51-148 wrocław, jurekp@microb.uni.wroc.pl piątkowski j., kliczkowska e.: influence of osmotic pressure on the growth of three species of the genus zoophthora. acta mycol. 43 (1): 49–55, 2008. strains accomodated in the genus zoophthora are very sensitive to osmotic value of their habitat. hipertonical molarity of buffers and nacl decreases the growth, but this effect strongly depends on the species tested and on the kind of buffer. in 0.66% phtalan buffer the growth of z. lanceolata is completely stopped whereas z. psyllae and z. aphrophora is inhibited only in 50% comparing to the control. key words: zoophthora, osmotic pressure, influence of buffers on growth introduction it is a well-established fact that many species of insects are infected and finally killed by fungi belonging to a variety of genera (batko 1964a, b; 1974). one of them is the genus zoophthora. investigations have been carried out into the use of entomopathogenic fungi as an alternative to chemical insecticides. however, entomopathogenic fungi are difficult to cultivate under laboratory conditions, so the researches have directed their attention to the factors that may influence the growth and germination of these species (freimoser et al. 1999) and thus facilitate their cultivation. the available information about the physical factors affecting the physiology of the entomopathogenic fungi is rather poor. research on a variety of physical factors has revealed that each species react differently to such factors. it has been noticed that the activity of entomopathogenic fungi against insects may depend on a season (bajan, kmitowa 1997; krejzowa 1988). although the seasons of the year themselves cannot be regarded as physical factors, we assume that the main factors that affect the physiological activity of the fungi in particular season are light, temperature and air humidity. many investigations into the influence of temperature have been reported so far (yendol 1968; hall, bell 1961). temperature is of importance to the growth of the acta mycologica vol. 43 (1): 49–55 2008 50 j. piątkowski and e. kliczkowska entomopathogenic fungi and to their ability of infecting insects (miętkiewski et al. 1994). in some instances the influence of temperature is positive (it stimulates), in others a negative one. at temperatures lower than 8°c and higher than 36°c, the spores of entomophthora virulenta are completely deactivated (yendol 1968). other investigations have revealed that the optimal temperature for the viability of the mycelium of particular fungal strain is also optimal for the viability of their spores (brobyn et al. 1985). the photoperiodic effect (feng, xu 1998) or the effect of ultraviolet light on some fungi has also been reported (braga et al. 1999). a significant factor which influences the fungal metabolism is moisture. as for the entomopathogenic fungi, this is also a very important factor of their habitat, responsible for viability, rate of growth and intensity of sporulation. for entomophthora thaxteriana, and e. aphidis the optimal moisture of spore release ranges from 70 to 90% at 20°c. as for the viability of the species, they may survive under these conditions for several days but in drier and colder air even for several weeks or months (wilding 1973). another important physical factor that affects the growth and sporulation of those fungi is light. exposure to electromagnetic waves in the visible range accounts for the effect of periodity of sporulation. the sporulation of most entomopathogenic fungi is light-dependend, but there are species that sporulate at a faster rate in darkness (latge et al. 1978). in our study reported on in this paper we cultivated three relative species of the genus zoophthora on the solid full medium supplemented with saline, phosphate buffer or phtalane buffer at different concentrations. the available literature contains no references to the importance of the osmotic value of the habitat to the physiological activity of fungi, involving the entomopathogenic strains. materials and methods the strains used in these tests: zoophthora aphrophorae (rostrup) bałazy, z. lanceolata keller and z. psyllae bałazy, were received from prof. stanisław bałazy, research centre for agricultural and forest environment, polish academy of sciences. z. aphrophora was isolated from aphrophora sp. the species was previously described by rostrup as enthomophthora aphrophorae and a second time by bałazy as zoophthora miridis (bałazy 1993). the name z. aphrophora has been proposed recently by prof. bałazy (bałazy 2003). z. lanceolata was isolated from empididae and z. psyllae from trioza urticae. all the strains were isolated in bavaria, germany. the strains were stored on the solid medium supplemented with a hen egg-yolk at 4°c. before testing, small samples of mycelium were translocated onto the solid full medium – ypg, and cultivated at 210c in the dark for 5 days. subsequently, small (7 mm diameter) discs of mycelium from the outer side of the cultivated fungus were cut out and translocated onto plates with solid full media (ypg) supplemented with buffers and/or nacl at concentrations of 0.0066m, 0.066m and 0.66m. use was made of phosphate and phtalan buffer according to the procedure described by mejbaum-katzenellenbogen and mochnacka (1968). we also used media with 0,0066m phosphate buffer supplemented with such an amount of nacl that the final nacl + buffer concentration amounted to 0.066m and 0.66m. each variant of the “strain+medium” was prepared in four repetitions, and statistical evaluation of the influence of osmotic pressure 51 diameter length was performed. air humidity at the time of culturing approached 55%. the media were prepared according to the following scheme: ______________________________________________________ components ____________medium number of medium i ii iii iv v vi vii viii ix x xi xii yeast extract [g] 10 10 10 10 10 10 10 10 10 10 10 10 peptone [g] 10 10 10 10 10 10 10 10 10 10 10 10 glucose [g] 20 20 20 20 20 20 20 20 20 20 20 20 agar-agar [g] 20 20 20 20 20 20 20 20 20 20 20 20 nacl [g] 0.38 3.85 38.6 3.47 34.7 phosphate buffer 0.66m ph=6.2 [ml] 1000 phosphate buffer 0.066m ph=6.2 [ml] 1000 phosphate buffer 0.0066m ph=6.2 [ml] 1000 1000 1000 acidic phthalate buffer 0.66m ph=6.2 [ml] 1000 acidic phthalate buffer 0.066m ph=6.2 [ml] 1000 acidic phthalate buffer 0.0066m ph=6.2 [ml] 1000 h2o [ml] 1000 1000 1000 1000 results and discussion figure 1 depicts the effect of osmothic prasure on the growth of the tree strains being tested (including statistically evaluated data). as we can see, at salt concentrations providing an isoosmotic environment of the medium-variant i (0.0066m), zoophthora psyllae grow at the fastest rate; the growth rate for z. aphrophora being slower and that of z. lanceolata the slowest one. in all the testing variants of the medium, the rate of mycelium growth of particular strain is similar in ypg (control, variant xii) and ypg suplemented with 0.066m buffer (iv, vii), or 0.0066m nacl (i). this implies that low concentrations of na2hpo4, kh2po4, nacl, and phthalate do not affect the rate of growth of the three fungal strains examined. if the concentration of the phthalate buffer or nacl amounts to 0.066m (ii,v), z. psyllae grows at the same rate as in the control, the diameter of the mycelium being 45.6 mm, 51.7 mm and 46.6 mm, respectively. in the x variant, when the phosphate influence of osmotic pressure 53 pseudocystidia are thought to be cells which perfore the cuticule of a host, enabling arising of conidiophores on outside of the host body (brobyn, wilding 1977). the morphologies of all these structures are of great importance to the differentiation of the genus zoophthora into particular species. but if we analyse the polymorphism and the extent to which the structures differ from one another, the difference will appear to be very smal in some instances. for example, on comparing the capillisporal structures as diagnostic features, one may conclude that the differences between the capillispore shapes are very subtile (keller 1980; ben-ze’ev, kenneth 1981b). when analysing the importance of morphological features to the diagnostics of fungal species, the question arises whether morphological differences are sufficient to distinguish particular species. in our opinion the analysis of morphological characteristics should be supported by the analysis of physiological properties to a greater extent than it is now. the aim of our study was not to revise the systematic classification of the species z. aphrophora, z. lanceolata or z. psyllae. more species representing each of the genera should be tested so that conclusions can be drawn about the taxonomic value of osmotic pressure. correct comparisons of the species require analysis at the molecular level – on the nucleotyde sequence in dna. but, in our opinion, such properties of the strains as sensitivity to osmotic preassure or to other physiological factors, may be also useful in differentiating those species into particular systematical units. the osmotic value of the fungal hyphe environment seems to be important, probably because of the exceptional sensitivity of the fungi to the decrease in water concentration inside the cells. the concentration of osmotically active substances in the fungal hyphe habitat, higher than the physiological ones accounts for the efflux of water from the inside of the cells. we woud like to emphasize that because of the high sensitivity of the fungi to osmotic value of the media used for their culturing is of great importance to the growth process, as well as to a variety of physiological processes, such as sporulation and release of secondary metabolites. but, as we can infer from the bars in figure 1, the responses to the varying buffer concentrations differ remarkably from one strain to another. the difference in sensitivity to the osmotic value of the environment between the fungal strains is also high, when compared the strains from not relative taxonomic units. for the yeast strains we used a storage medium, where one of the components, glycerol, accounted for 50% of the total content. in such medium the yeast cells can survive at temp. 710c for years, except the zoophthora species, which survived only one day, when kept in this environment (results not published). it is not clear why some fungal taxa are so sensitive to water amount. particular physiological processes involved in this “water-dependence” have not been well recognized so far. probably, some enzymes in “low water-sensitive” strains change their conformation, especially at an active centre, when water concentration changes. the level of this sensitivity varies from organisms to organisms. water can be removed completely from yeast and bacteria, but they recover life activity in the water habitat. these differences in viability under low moisture conditions may be of great ecological importance. 54 j. piątkowski and e. kliczkowska the use of a media differing in osmotic values may be beneficial when detecting and investigating biochemical changes in fungal cell metabolism with the decrease in the water level. it is worth noticing that the buffers and nacl not only decrease the growth of the strains but also the morphology (colour) of particular fungal colony is different, depending to the medium variant, as illustrated in the figure 2a, b. the medium variants were prepared according to the procedure shown in the schema in materials and methods. to sum up, the results suggest that osmotic pressure is one of the significant factors that affect growth and presumably other physiological processes of entomopathogenic fungi belonging to the genus zoophthora. it is obvious that other comparative investigation must be performed, using other species of this kind of fungi. references bałazy s. 1993. flora polska. grzyby (mycota) 24:owadomorkowe (entomophthorales). w. szafer institute of botany, polish academy of sciences, kraków. bałazy s. 2003. on some little known epizootics in noxious and beneficial arthropod populations caused by entomophthoralean fungi. insect pathogens and insect parasitic nematodes iobc bulletin 26: 63–68. batko a 1964a. remarks on the genus entomophthora. bull. acad. pol. ser. sci. biol. 12: 319–321. batko a. 1964b. on the new genera zoophthora. bull. acad. pol. ser. sci. biol. 12: 323–326. batko a. 1974. filogeneza a struktury taksonomiczne entomophthoraceae (phylogenesis and taxonomic structure of entomophthoraceae). (in:) c. nowiński (ed.). ewolucja biologiczna, szkice teoretyczne i metodologiczne. polska akad. nauk, ossolineum, wrocław: 109–303. ben-ze’ev i., kenneth r. g. 1981. zoophthora radicans and zoophthora petchii sp. nov. (zygomycetes: entomophthorales) two species of the “sphaerosperma – group” attacking leafhoppersand froghoppers (hom.). entomophaga 26: 131–142. braga g., messias c. l., miller c. d., flint s. d., daldwell m., anderson a. j., roberts d. w. 1999. effect of near ultraviolet light (uva and uvb) on hydrated and germinating conidia of the genus metarhizium. xxxiii annual meeting of the sip, irvaine. sip program and abstracts: 28. bajan c., kmitowa k. 1997. interactions between entomopathogenic and saprophytic fungi. zar ja. bio-interactions between entomopathogenic and saprophytic fungi. zar ja. biostatical analysis. 32: 47–63. brobyn p. j., wilding n. 1977. invasive and developmental processes of entomophthora species infecting aphids. trans. brit. mycol. soc. 69: 349–366. brobyn p. j., wilding n., clark s. j. the persistence of infectivity of conigia of the aphid pathogen, erynia neoaphidis on leaves in the field. ann. appl. biol. 107: 365–376. feng m. g., xu j. h. 1998. photoperiods affect the growth and sporulation pattern of the liquid and plate cultures of the entomophthoralean fungus pandora delphacis (hori) humber. (in:) viith international colloquium on invertebrate pathology, sapporo. meetings program and abstracts: 55. freimoser m., grundschoher f. aebi m., tuor u. 1999. morphology and growth requirements of the thrips pathogenic fungus enthomophthora thripidum in in vitro cultures. (in:) xxxiii annual meeting of the sip, irvaine. sip program and abstracts: 36. hall i. m., bell j. v. 1961. further studies on the effect of temperature on the growth of some entomophthoraceus fungi. journal of insect pathology 3: 289–296. keller s. 1980. two new species of the genus zoophthora batko (zygomycetes, entomophthorales): z. lanceolata and z. crassitunicata. sydowia 33: 167–173. latge j.p., perry d., papierok b., coremans-pelseneer j., remaudiere g., resinger o. 1978. germination d’ azygospore d’ entomophthora obscura. acad. sci. paris ser. 287: 946. mejbaum-katzenellenbogen w., mochnacka i. 1968. kurs praktyczny z biochemii. pwn warszawa. wilding n. 1973. the survival of entomophthora spp. in mummified aphids at different temperatures and humidities. j. invertebr. pathol. 21: 309–311. yendol w.g. 1968. factors affecting germination of entomophthora conidia. j. invertebr. pathol. 10: 116–121. influence of osmotic pressure 55 wpływ ciśnienia osmotycznego na wzrost trzech gatunków z rodzaju zoophthora s t r e s z c z e n i e informacje o wpływie czynników fizycznych na grzyby owadobójcze są w literaturze naukowej raczej skąpe. donosi się o znaczeniu temperatury, wilgotności powietrza, oraz długości działania światła na wzrost i sporulację wybranych gatunków. w niniejszej pracy przedstawiono wyniki badań nad wpływem różnych stężeń buforu fosforanowego, ftalanowego oraz chlorku sodowego na wzrost szczepów trzech gatunków: zoophthora lanceolata, z. psyllae i z. aphrophora. stężenie buforów i chlorku sodowego, które można uznać za zbliżone do izoosmotycznego w stosunku do komórek grzybów (0.0066m), nie zmienia szybkości wzrostu szczepów w porównaniu do kontroli. stężenia dziesięć razy większe spowalniają wzrost, szczególnie w stosunku do z. lanceolata i przy buforze fosforanowym. silniej hamuje bufor fosforanowy niż ftalanowy, co może oznaczać, iż jony fosforanowe zwiększają wrażliwość tych grzybów na podwyższone ciśnienie osmotyczne. stężenie 100 razy większe od izoosmotycznego prawie całkowicie hamuje wzrost, ale w obecności buforu ftalanowego obserwuje się znaczną różnicę we wrażliwości między z. lanceolata a dwoma pozostałymi szczepami, co stwarza nadzieję, iż zachowanie się poszczególnych szczepów gatunków z rodzaju zoophthora w obecności podwyższonych stężeń buforów może mieć znaczenie diagnostyczne. i iii v ii iv vi 1 1 1 1 1 1 2 2 2 2 2 2 3 3 3 3 3 3 fig. 2a. morphology and size of the mycelia of z. aphrophora, z lanceolata and z. psyllae on media with different concentrations of buffers and nacl. vii ix xi viii x xii 1 1 1 1 1 1 2 2 2 2 2 2 3 3 3 3 3 3 fig. 2b. morphology and size of the mycelia of z. aphrophora, z. lanceolata and z. psyllae on media with different concentrations of buffers and nacl. 2014-01-01t11:47:11+0100 polish botanical society ecology and plectology of phlebia tremelloidea (polyporales, agaricomycetes) ivan v. zmitrovich1 and oleg n. ezhov2 1v.l. komarov botanical institute, 2 popov street, ru-197376 st. petersburg, russia iv_zmitrovich@mail.ru 2institute of ecological problems of the north, 23 north dvina quay ru-163000 arkhangelsk, russia, elegezhik@gmail.com zmitrovich i.v., ezhov o.n.: ecology and plectology of phlebia tremelloidea (polyporales, agaricomycetes). acta mycol. 46 (1): 19–25, 2011. a rare boreonemoral species, phlebia tremelloidea (bres.) parmasto was characterized morphologically and ecologically basing on russian material. the specified description of the species was given. the variability of top lamprocystidia, basidia and basidiospores of the fungus was revealed. an abhymenial, medullar, and subhymenial strates of the basidiocarp were characterized. the relationships between developmental environments and morphology of the fungus were discussed. key words: basal layer, boreonemoral forests, ecology, gelatinized basidiocarps, lamprocystidia, medullar layer, phlebioid fungi, slowly-growing resupinates, thickened hymenium introduction phlebia tremelloidea (bres.) parmasto [= ph. lindtneri (pilát) parmasto] is rare boreonemoral species known from several localities on eurasian continent. rather variable morphology of this species is a reason of its controversial descriptions as well as rich synonymy, sound for rare taxon. a new finding of this fungus in old boreal forest of arkhangelsk region (european russia) feats us re-examine all accessible material on the species. therefore, the purpose of the present note is generalization of data on taxonomy, morphology and ecology of ph. tremelloidea. acta mycologica vol. 46 (1): 19–25 2011 20 i.v. zmitrovich and o.n. ezhov material and methods all accessible material on ph. tremelloidea, phlebia lindntneri, peniophora tremelloidea bres., and phlebia merulioidea parmasto were involved into present investigation. the collections consist of material collected by the authors in some regions of russia as well as selected specimens from the mycological herbarium of the komarov botanical institute (le) (saint petersburg). all specimens were studied and preserved with standard methods. the macroscopic description was based on examination of the fresh material and the analysis of the photos. the dried material was studied using light microscopic techniques. microscopical study of basidiocarps was carried out as described by gilbertson and ryvarden (1986). freehand sections and squash mounts of basidiocarps were examined in 5% koh and 2% cotton blue. spore conglomerations were searched in hymenophoral gelatinose matrix. spore measurements contained 30 spores per specimen. the following abbreviations were used: l – spore length, w – spore width, q – quotient of the spore length and spore width (l/w ratio). specimens are preserved in the herbarium of komarov botanical institute (st. petersburg, le) and institute of ecological problems of the north (arkhangelsk, ar). results species description phlebia tremelloidea (bres.) parmasto, eesti nsv tead. akad. toim., biol. seer 16: 393, 1967. ≡ peniophora tremelloidea bres., ann. mycol. 18: 48, 1920. = p. lindtneri pilát, bull. trimest. soc. mycol. france 53: 97, 1937. ≡ phlebia lindtneri (pilát) parmasto, wahlenbergia 1: 74, 1975. ≡ hohenbuehelia lindtneri (pilát) spirin in zmitr., v. malysheva, e. malysheva & spirin, folia cryptogamica petropolitana 1: 71, 2004. = phlebia merulioidea parmasto, notul. syst. sect. cryptog. inst. bot. acad. sci. urss 15: 127, 1962. basidiocarps annual to persisting, resupinate, widely effused, originated as orbicular patches 1-4 cm in diam. and 0.3-3 mm thick, which merge into continuous crusts up to 20 cm long, subhyaline, honey-yellow or pale-creamish with gray to tan tints. the consistency is tough gelatinose when fresh, hard corneous when dry. the margin free laying, 0.2-0.8 mm wide, radially fibrillose to ciliate, in inrolled parts squalid due to weakly differentiated cilia, of the same consistency and color as basidiocarp. hymenophore gyrose, 0.2-1 mm thick, of typical merulioid appearance, gelatinose to corneous of the same color as basidiocarp, but with shine pruina due to free-leaving hymenial lamprocystidia; alveoles 0.3-0.5(2) mm wide (fig. 1). fig. 1. aspects of basidiocarp of phlebia tremelloidea (fresh material; le 269621) highlighting hymenophoral merulioid folds. scale bar = 1 mm (photo r.v. ershov). fig. 2. transverse section through basidiocarp of phlebia tremelloidea: bl – basal layer with coiled agglutinated hyphae (ahc – agglutinated hyphae of cuticular cover); ml – agglutinated medullar hyphae (amh) producing a mucilaginous secret (gray phone) which forms cartilaginous matrix of basidiocarp; sh – subhymenium, composed by densely packed candelabriform hyphae; h – current hymenium with basidia (b), terminations of lamprocystidia (l) and basidiospores (bs). scale bar = 10 μm (fresh material; le 269621). ecology and plectology of phlebia tremelloidea 21 hyphal system monomitic. hyphae fibulate, hyaline, 1.5-3(7) μm in diam., with strongly gelatinized walls; in subhymenium 2-3 μm in diam, densely packed with formation of obscure textura porrecta; in medullar strate 2-5(7) μm free merged into gelatinose matrix, regularly branching, often inflated or coiled; near abhymenial surface 1.5-4 μm in diam., coiled and densely packed into superficial fascicles, strongly agglutinated. lamprocystidia originating from low medullar to hymenial and abhymenial layers, abundant (predominating in fold areas), 60-120 × (5)9-12(18) μm (encrusted part varying between 38-60 × 8-18 μm; tab. 1), subulate to fusoid, initially naked and with slightly thickened walls, in maturity thick-walled and heavily encrusted in the top third (ecncustation pyramidal crystalline, oxalate). basidia long-cylindrical, 31-42 × 3.9-5.1 μm, 4-spored, with weak constriction and a clamp at the base (tab. 1). basidiospores 4.8-7.2 × 2.5-4.1 μm, ellipsoid to ovoid, slightly flattened, thin-walled, inamyloid, hyaline, with numerous small droplets (tab. 1, fig. 2). table 1 variability of diagnostically important microstructures in phlebia tremelloidea (fresh material le 269621) encrusted parts of lamprocystidia basidia basidiospores l w q l w q l w q 42 11 3.8 32 3.9 8.2 5.2 2.9 1.8 48 11 4.4 39 4.1 9.5 5.4 3 1.8 38 10 3.8 35 4.5 7.8 5.5 3.9 1.4 52 15 3.5 31 4 7.8 4.9 3.2 1.5 58 13 4.5 42 5 8.4 7.2 3.9 1.8 45 12 3.8 31 4.5 6.9 5 2.9 1.7 50 18 2.8 35 4.6 7.6 5.3 3 1.8 58 15 3.9 32 3.9 8.2 5.1 2.6 2.0 60 18 3.3 35 4.9 7.1 5.4 2.8 1.9 48 12 4.0 32 4 8.0 7.8 4.1 1.9 45 10 4.5 41 4.5 9.1 4.8 2.5 1.9 56 12 4.7 42 5.1 8.2 5.2 3 1.7 62 17 3.6 35 5 7.0 5.7 2.8 2.0 45 15 3.0 32 4 8.0 5.1 2.8 1.8 38 10 3.8 30 3.9 7.7 5.2 2.7 1.9 47 13 3.6 32 4.6 7.0 6.5 3.5 1.9 42 14 3.0 33 3.9 8.5 5.9 2.9 2.0 38 9 4.2 38 4 9.5 5.2 3 1.7 35 8 4.4 42 5 8.4 5.2 2.5 2.1 45 10 4.5 38 4.2 9.0 4.8 2.5 1.9 48 12 4.0 42 4.7 8.9 6.2 3.2 1.9 39 10 3.9 40 5 8.0 5.1 3.1 1.6 45 15 3.0 33 4.5 7.3 6.5 3.1 2.1 58 16 3.6 33 3.9 8.5 7 4.1 1.7 39 10 3.9 35 4 8.8 5.3 3.2 1.7 55 15 3.7 38 4.1 9.3 5.2 2.8 1.9 45 13 3.5 42 5 8.4 6 3.5 1.7 38 10 3.8 39 3.9 10.0 5.5 3.2 1.7 55 14 3.9 40 4.5 8.9 5.8 3.2 1.8 42 12 3.5 38 4.2 9.0 5.2 2.8 1.9 47.2 12.7 3.8 36.2 4.4 8.3 5.6 3.1 1.8 abbreviation. the limiting dimensions marked by boldface. 22 i.v. zmitrovich and o.n. ezhov on bark or fully decorticated areas of fallen logs on abies sibirica, picea abies, alnus incana, populus tremula, and quercus robur in old boreal or boreonemoral forests. associated with a white rot. specimens examined. russia: arkhangelsk region: koryazhma vicinities, oxalis acetosella spruce-fir forest, on bark of fallen abies sibirica, 9.viii.2010, leg. o.n. ezhov (le 269621). leningrad region: vsevolozhsk district, vaskelovo vicinities, aegopodium podagraria alder forest, on bark of fallen alnus incana, 27.viii.1997, leg. i.v. zmitrovich (le 214796; as phlebia lindtneri). nizhegorod region: kerzhensky reserve, floodland of kerzhenetz, on decorticated fallen log of quercus robur, 10.x.1999, leg. w.a. spirin (le 208542; as phlebia lindtneri). krasnoyarsk region: enisey district, kolchym vicinities, fir forest, on fallen decorticated populus tremula, 14.viii.1959, leg e. parmasto (le 22527; isotype of phlebia merulioidea). discussion taxonomical history. the type species was described by bresadola (1920) in genus peniophora cooke due to its striking encrusted lamprocystidia and hard hymenium with thickened subhymenieum. some years later, pilát described his peniophora lindtneri (pilát 1937) sufficiently corresponded to bresadolian p. tremelloidea. parmasto (1967) shifted p. lindtneri into genus phlebia fr. on the reason of its gelatinized textura and more appropriate to phlebia characteristics of cystidia, basidia, and basidiospores. in such a status, the name was in wide use (eriksson et al. 1981; jülich, stalpers 1980; kotiranta, saarenoksa 2000). parallelly, parmasto described phlebia merulioidea (parmasto 1962) for the fungus sufficiently corresponded to ph. tremelloidea and ph. lindtneri. as a difference of newly described species from ‘peniophora tremelloides’, this author mentioned its narrower thin-walled hyphae (1.4-4.5 vs 4-7 μm) and smaller basidiospores (4.5-5.5 × 2.3-2.8 vs 6-7 × 4-5 μm). the further investigations of microstructures of both species fall the border between these (see tab. 1). the position of the species within genus phlebia is rather stable and widely accepted. only in 2004, it was made new combination hohenbuehelia lindtneri (pilát) spirin (zmitrovich et al. 2004) on the base of metuloid-like lamprocystidia and a local attachment area of basidiocarp, corresponded with consideration on merulioid fungi as resupinate agarics with disturbed morphogenesis. however, strict phlebioid textura of basidiocarp has forced a re-evaluation of this position for the benefit of recognizing a separate unit of generic rank, lampromerulius zmitr. & spirin ined. (spirin, zmitrovich 2009). the recent molecular study of phlebioid fungi (moreno et al. 2010) based on divergence revealing by its and partial 28s rdna sequences not supported an isolated position of ‘lampromerulius’: the sequence ph. lindtneri was clustered with those of ‘steccherinum’ lusitanicum (bres.) ryvarden and phlebia setulosa (berk. & m.a. curtis) nakasone in core-phlebia clade. therefore, the lamprocystidia presence and construction is trailer apomorphy needed in ecomorphological rather than phylogenetical interpretation. plectology. the hyphae of ph. tremelloidea produce mucilaginous substance that leads to cartilaginous context of basidiocarp. such a feature is connected to ecology and plectology of phlebia tremelloidea 23 reduction of water loss by evaporation and protection the protein bearing hyphae of the deeper regions against microscopic animals, such as mites and spring tails. gelatinized contexts represent a certain water reserve and give important volume to a basidiocarp without engaging much biomass (clémençon 2004). the basal layer of basidiocarp of ph. tremelloidea represents an agglutinated hyphal mass forming dense abhymenial plate, i.e. this layer cannot be interpreted as subiculum, rooted into substrate (fig. 2). the hyphae in this layer are variable in diameter (1.5-4 μm wide) and often bear thickened walls that more precisely can be interpreted as prominent glucane cover. usually, under 5% koh these thick-walled hyphae dissolve and stay subinvisible. the hyphal arrangement in this layer is more or less horizontal. near the margin the hyphae form ciliar agglutinated fascicles, furnished by occasional lamprocystidia. the medullar layer of basidiocarp in ph. tremelloidea is weakly differentiated from upper and lower strates. this is a huge cartilaginous mass composed by freeleaving strongly gelatinized hyphae. as it seen on figure 2, the hyphal mass of this layer is subinvisible due to mucilaginous substance. the hyphae in this layer have descending orientation. near to subsequent subhymenial strate, some hyphae bear fusiform apical thick-walled swellings. these are precursors of lamprocystidia. in general, hyphal masses of medullar layer form a certain fascicular zones corresponded to hymenophoral ridges. the hyphal ends in these ridges have a tendency to lamprocystidia formations. the fungus in question is characterized by thickened hymenium (auxohymenium according to clémençon 2004), i.e. candelabriform structure leading to textura porrecta, which is a result of replacement of old basidia by new ones springing from ramifications of the same candelabra growing beyond the general level of the hymenium. in ph. tremelloidea this layer is enriched and strengthened by mature lamprocystidia. the roots of these strucures are hidden among subhymenial textura porrecta, but in some cases we can mark a total length which reaches 120 μm. however, encrusted part, which consists as a rule 1/3 of total length, is easily observable, and varies in limits 38-60 × 8-18 μm (tab. 1). the current hymenium of ph. tremelloidea composed by basidia, lamprocystidial ends and small young lamprocystidia (fig. 2). basidia of characteristic for phlebia type – an elongated, cylindrical with weak constriction and clamp at the base, the limits of their variability 31-42 × 3.9-5.1 μm (tab. 1). agglutinated area of hymenial surface bear a lot of ellipsoid and slightly flattened thin-walled basidiospores 4.8-7.2 × 2.5-4.1 μm (tab. 1, fig. 2). ecology. ph. tremelloidea is sparsely distributed in forest zone of western and north europe (bresadola 1920; pilát 1937; ryvarden et al. 1981; hansen, knudsen 1997; kotiranta, saarenoksa 2000) and european (spirin 2002; zmitrovich et al. 2004) and asian (parmasto 1962, 1968) russia. as preferred communities were mentioned fir forest (parmasto 1962), pine and spruce-mixed forests (ryvarden et al. 1981), juniper-mixed herb-rich spruce forest (kotiranta, saarenoksa 2000), nemoral floodland (spirin 2002), and herb-rich alder forest (zmitrovich et al. 2004). our new finding in arkhangelsk region was made in mixed spruce-fir forest (picea abies, abies sibirica) of diverse age with some intrusions of pine (pinus sylvestris), birch (betula pubescens) and aspen (populus tremula). an average age of 24 i.v. zmitrovich and o.n. ezhov stand consists 160–180 years, the spruce actively falls, therefore there are some clear areas enriched by gross wood-debris. the soil cover is presented by oxalis acetosella, vaccinium myrtillus, hylocomium splendens, dryopteris filix-mas. this type can be characterized as piceeto-abiegnetum meso-hygrophilico herbosum. the fungus was attached to shady site of community. other members of local fungal community are oligoporus immitis (pers.) niemelä (aspen), amylocystis lapponicus (romell) bondartsev & singer, phellinus chrysoloma (fr.) donk, phlebiopsis gigantea (fr.) jülich, pycnoporellus fulgens (fr.) donk (spruce), phellinus pini (brot.) bondartsev & singer, skeletocutis carneogrisea a. david (pine), oligoporus rancidus (bres.) gilb. & ryvarden (fir). concerning substrata, the fungus was reported from populus tremula (parmasto 1962), picea abies and pinus sylvestris (ryvarden et al. 1981), juniperus communis (kotiranta, saarenoksa 2000), quercus robur (spirin 2002) and alnus incana (zmitrovich et al. 2004). our finding in arkhangelsk region is associated with fallen abies sibirica, diameter of log reaches 28 cm. it is necessary to mention that material on populus tremula, quercus robur, and alnus incana is associated with strictly decorticated areas, whereas our material on abies sibirica is associated with uncracked bark. the fungal outgrowths appear from underside of fallen logs as roundish patches having an intermerging tendency. the margin generally adhere the substrate, but without rooting into them. some specimens demonstrate inrolled margin with naked or pruinose (on the manner of phlebia centrifuga p. karst.) abhymenial surface. the fact that fully developed basidiocarps can be revealed in august (parmasto 1962; zmitrovich et al. 2004; the present data) and keep up to october (kotiranta, saarenoksa 2000; spirin 2002) (and, possibly, later) forces us suppose that the fungus is a slowly growing with potential to survive of winter season. the clearly thickened subhymenium protruded by lamprocystidia supports parmasto (1962) interpretation of hymenium of the fungus as ‘multilayered’. the observations of the fungus during some years are needed to revealing of persisting capacity of this species. conclusion ph. tremelloidea is a slowly-growing phlebioid fungus adapted to colonization of homogeneous (not cavernose) substrata, like decorticated wood or uncracked bark. in contrast to relatives having a transcortical development, like ph. radiata fr. or ph. rufa (pers.) m.p. christ., this fungus haven’t reserved superficial vegetative mass. therefore, its growth is more slow and even. the numerous lamprocystidia create a rigid skeleton which promotes to constant increasing of hymenial field. the fungus attached to soft conditions of old forests with abundance of fallen logs that allows a long-term (to persisting) sporulation. acknowledgements. the authors are very grateful to dr. r.v. ershov for discussions on this interesting species and the photos. financial support from rfbr grants “regularities of mycobiota formation of boreal forests on north-west of russia in cretaceous landscapes environments” (n 08-04-98805-р_north_а) and “substrate preference and spatial structure of biota of aphyllophoroid fungi in nature ecosystems of european russia” (09-04-01064-а). ecology and plectology of phlebia tremelloidea 25 references bresadola g. 1920. selecta mycologica i. ann. mycol. 12: 26–70. clémençon h. 2004. cytology and plectology of the hymenomycetes. bibl. mycol. 199: 1–488. eriksson j., hjortstam k., ryvarden l. 1981. the corticiaceae of north europe. 6. phlebia – sarcodontia. fungiflora, oslo: 1051–1276. gilbertson r. l., ryvarden l. 1986. north american polypores. 1. fungiflora, oslo, 436 pp. hansen l., knudsen h. (eds). 1997. nordic macromycetes. 3. heterobasidioid, aphyllophoroid and gastromycetoid basidiomycetes. nordsvamp, copenhagen, 445 pp. jülich w., stalpers j.a. 1980. the resupinate non-poroid aphyllophorales of the northern hemisphere. north-holland pub. comp., amst.; oxf.; new york, 335 pp. kotiranta h., saarenoksa r. 2000. corticioid fungi (aphyllophorales, basidiomycetes) in finland. acta bot. fennica 168: 1–55. moreno g., blanco m.-n., platas g., peláez f. 2010. taxonomic and phylogenetic revision of three rare irpicoid species within the meruliaceae. mycol. progress 10 (a preprint). parmasto e. 1962. species varietatesque novae fungorum. tremellales et aphyllophorales. notulae systematicae e sectione cryptogamica instituti botanici nomine v.l. komarovii academiae scientiarum urss 15: 125–137. parmasto e. 1967. corticiaceae u.r.s.s. iv. descriptiones taxorum novarum. combinationes novae. eesti nsv tead. akad. toimet. biol. 16: 377–394. parmasto e. 1968. conspectus systematis corticiacearum. inst. zool. bot., tartu, 261 pp. pilát a. 1937. addimenta ad floram sibiriae asiaeque orientalis mycologicam. pars iv. bull. soc. mycol. france 52: 305–336. spirin w.a. 2003. aphyllophoraceous fungi of nizhegorod region: species composition and ecological peculiarities. phd thesis. komarov botanical institute, st. petersburg, 242 pp. spirin w.a., zmitrovich i.v. 2009. phaeophora: new taxa compendium. komarov botanical institute, st. petersburg. 1 s. (in latin, a manuscript). zmitrovich i.v., malysheva v.f., malysheva e.f., spirin w.a. 2004. pleurotoid fungi of leningrad region (with notes on rare and interesting east european taxa). folia crypt. petropolitana 1: 1–124. 2014-01-01t11:51:50+0100 polish botanical society association of water-borne conidial fungi with epiphytic tree fern (drynaria quercifolia) kishore s. karamchand and kandikere r. sridhar microbiology and biotechnology, department of biosciences mangalore university, mangalagangotri, mangalore in-574-199, sirikr@yahoo.com karamchand k. s., sridhar k. r.: association of water-borne conidial fungi with epiphytic tree fern (drynaria quercifolia). acta mycol. 44 (1): 19–27, 2009. the live and dead tissues, and trapped leaf litter by the epiphytic tree fern drynaria quercifolia associated with riparian tree species of konaje (west coast) and sampaje (western ghat) streams of india during dry (summer) and wet (monsoon) seasons yielded 37 species of water-borne conidial fungi on bubble chamber incubation. dead bracket leaves of fern possess the highest species as well as conidia in konaje, while the trapped leaf litter in sampaje. during summer, the diversity was highest in bracket leaves in both locations, while in monsoon season it was highest in rhizomes of konaje and in trapped leaf litter in sampaje. even though the conidial output from tissues of drynaria and trapped leaf litter were not equivalent to stream submerged leaf litter, the species richness ranged between 40% and 75% in konaje and sampaje streams. as stable epiphyte, drynaria exposed to wet and dry regimes in tree canopies of west coast and western ghats likely to serve as host for perfect states of water-borne hyphomycetes. key words: water-borne hyphomycetes, diversity, conidia, canopy, drynaria, epiphyte, tree fern introduction aquatic hyphomycetes (also known as ‘freshwater fungi’ and ‘ingoldian fungi’) are phylogenetically heterogeneous group dominant on submerged leaf litter in streams (bärlocher 1992). in addition to aquatic hyphomycetes, aero-aquatic hyphomycetes are also known from dead plant litter in streams (e.g., helicosporus fungi). aquatic hyphomycetes produce conidia with three basic shapes (conventional, sigmoid and multiradiate) (ingold 1942, 1975a; goos 1987), while helicosporus fungal spores are twisted at least up to 180° (goos 1987). the sigmoid and multiradiate shapes of aquatic hyphomycetes are the product of convergent evolution for adaptation in flowing waters (ingold 1975b). helicosporus fungi also showed convergent lineages and their spores evolved for dispersion in water by trapping air-bubbles (zhao et al. 2007). aquatic hyphomycetes and helicosporus fungi have been connected to their acta mycologica vol. 44 (1): 19–27 2009 20 k. s. karamchand and k. r. sridhar perfect states by conventional (webster, descals 1979; goos 1987; webster 1992; sivichai, jones 2003; zhao et al. 2007) and molecular techniques (belliveau, bärlocher 2005; campbell et al. 2006; tsui, barbee 2006). besides aquatic habitats, water-borne hyphomycetes are also known from terrestrial habitats (e.g., soil, leaf litter, roots, leaf surfaces) (sridhar, bärlocher 1993; bärlocher 2006). forest canopies represent a broad group of fungi (e.g. endophytic, pathogenic, phylloplane and lignicolous fungi (lodge, cantrell 1995). water-borne hyphomycetes are also common in tree canopies such as throughfall, stemflow, tree holes and epiphytes of temperate regions) (canada, hungary, japan and poland) (bandoni 1981; ando, tubaki 1984; czeczuga, orłowska 1998; gönczöl, révay 2006). recently, they have also been reported from the epiphytic fern and tree holes in india (sridhar et al. 2006; karamchand, sridhar 2008). heavy rainfall (350–650 cm/annum) during southwest monsoon (during june-september) results in continuous wet conditions in tree canopies of the western ghats and west coast of southern india. it is likely such canopies support the growth, sporulation and dissemination of water-borne hyphomycetes. tree ferns with their overlapping fronds access nutrients from stem flow, through fall and drips, and serve as ‘trash baskets’. the tree fern drynaria quercifolia, also known as ‘oak-leaf basket fern’ is a dominant fern in western ghats and west coast of india. their bracket leaves serve as natural funnels to trap canopy debris (e.g. flowers, dead bark, dead leaves). in view of a few studies on water-borne hyphomycetes in tree canopies in tropics (sridhar et al. 2006; karamchand, sridhar 2008), the present study aims at evaluating their assemblage and diversity in d. quercifolia associated with riparian trees of the west coast and western ghat region during summer and monsoon seasons. materials and methods five riparian trees species along the konaje stream (90 m a.s.l.; 12°48′n, 74°55′e) of the west coast and sampaje stream (510 m a.s.l.; 12°29′n, 75°35′e) of the western ghats associated with drynaria quercifolia (linn.) j. sm. (polypodiaceae) were chosen for the study. this tree fern produces two kinds of leaves: nest leaves (slender, fragile, short-lived, turn brown on senescence and shed on drying with persistent rachis) and bracket leaves (tough, leathery, resembling oak leaves, remain attached to creeping rhizome during senescence and dried condition, and decay in standing dead position). the distance between the stream and trees sampled was between 2 and 50 m, while the height at which the fern attached to trees was between 2 and 20 m. during february (summer) and july (monsoon) 2006, from each fern, pieces of rhizome with roots, partially skeletonized bracket leaves and trapped leaf litter were sampled in sterile polythene bags and transferred to the laboratory. randomly excised tissues (5-6 leaf and rhizome pieces, 1×2.5 cm; 15-20 small root pieces) were rinsed in water to eliminate surface debris, suspended in sterile distilled water in five replicates (each in 250 ml erlenmeyer flask) and bubbled through pasteur pipette using aquarium aerators (48 hr, 23±2°c). aerated water was filtered (millipore filters, 5 µm), stained with 0.1% cotton blue in lactophenol and enumerated the liberated conidia from each substrate and calculated the conidial association of water-borne fungi 21 output per gram dry mass of substrate. the mean conidia produced by each fungus per mg dry mass were also calculated: mean conidia/fungus/mg dry mass = (total conidia liberated/mg) ÷ (total species recorded) the simpson (d’) and shannon (h’) diversities (magurran 1988) in each substrate were estimated: simpson’s index: d′ = 1 ÷ ∑ (pi)2 shannon index: h′ = – ∑ (pi ln pi) (where, pi is the proportion of fungal species i contributes to the total). the evenness (j’) fungi in different substrates (pielou 1975) were expressed: j′ = h′ ÷ h′max (where h′max is the maximum value of diversity for the number of fungal species present). percent jaccard’s index of similarity (ji) was calculated for the fungi colonized in different substrates pair-wise (kenkel, booth 1992): ji (%) = (c) ÷ (a + b + c) × 100 (where, c is the number of fungal species occurring in both substrates, a is the number of fungal species unique to the first substrate and b is the number of fungal species unique to the second substrate). paired t-test was employed to assess the difference in conidial output from different substrates between summer and monsoon seasons (parameters: p value, two tailed; confidence interval, 95%) (statsoft 2008). results and discussion a total of 24 species of water-borne hyphomycetes was associated with tissues of drynaria and trapped leaf litter in konaje region (tab. 1). the species richness was table 1 water-borne hyphomycetes (conidia/g dry mass) recovered from tissues of drynaria and trapped leaf litter in riparian trees of konaje region during summer and monsoon (in parenthesis) (mean, n = 5) (different letters across the total conidia of each substrate between summer and monsoon indicate significant difference; p < 0.05; paited t-test) species bracket leaf root rhizome trapped leaf anguillospora crassa ingold 56 (7) 63 (6) 4 (6) 5 a. longissima (sacc. & p. syd.) ingold 301 (7) 83 (16) (17) 2 (638) arborispora sp. 5 (5) (8) campylospora chaetocladia ranzoni 18 (103) clathrosphaerina zalewskii beverw. (7) cylindorcarpon sp. 20 153 10 1 (8) flabellospora crassa alas. 42 f. verticillata alas. 5 (3) 1 f. multiradiata nawawi (3) flagellospora curvula ingold 319 17 78 (218) 22 k. s. karamchand and k. r. sridhar f. penicillioides ingold 723 2 4 helicosporium sp. 8 lunulospora curvula ingold (2) magdalaenaea monogramma g. arnaud (29) phalangispora constricta nawawi & j. webster (2) retiarius bovicornutus d.l. olivier 8 1 tetracladium marchalianum de wild. (5) tricladium malaysianum nawawi 18 5(2) (1) 3 trinacrium subtile riess 208 tripospermum infalcatum k. ando & tubaki 8 triscelophorus acuminatus nawawi 36 (100) 13 (4) (7) 2 (144) t. konajensis k.r. sridhar & kaver. 8 (39) 5 (2) (3) 3 (284) t. monosporus ingold (3) (2) tumularia aquatica (ingold) descals & marvanová (2) (3) total species 15 (7) 8 (9) 3 (9) 10 (8) cumulative species 15 (7) 16 (11) (16) 14 16 (18) total conidia/g dry mass 1778a (166b) 344a (38b) 16a (50a) 100a (1432b) fig. 1. mean number of species (a) (n=5, mean±sd) and mean conidia (n=5) per species per gram dry mass (b) of bracket leaf, root, rhizome (drynaria quercifolia) and trapped leaf litter of konaje region. tab. 1. cont. association of water-borne fungi 23 highest in bracket leaves and lowest in rhizomes (15 vs. 3 species) during summer. the conidial output was peaked in bracket leaves and lowest in rhizomes during summer (1778 vs. 16/g). the cumulative species between substrates of drynaria and trapped leaves was more or less constant during summer (15-16), while it steadily increased during monsoon (7-18) (tab. 1). the mean number of species and mean conidia per species were highest in bracket leaves in summer followed by trapped leaves in monsoon (fig. 1). tissues of drynaria and trapped leaves of sampaje region yielded 27 species (tab. 2). the trapped leaves possess highest number of species during summer as well as monsoon (15 vs. 13 species) (tab. 2). the total conidial output was highest in trapped leaves during summer (363/g). the cumulative species between substrates of drynaria and trapped leaves was steadly increased in monsoon (7-18) as well as summer (5-15) seasons. the mean conidial output per species was highest in roots during summer (fig. 2). significant difference (paired t-test) in conidial out put was seen between summer and monsoon in bracket leaves (p = 0.0319), roots (p = 0.0424) and trapped leaves (p = 0.0336) in konaje region, while there was no significant difference in sampaje region. the fungal diversity was table 2 water-borne hyphomycetes (conidia/g dry mass) recovered from tissues of drynaria and trapped leaf litter in riparian trees of sampaje region during summer and monsoon (in parenthesis) (mean, n = 5) (same letters across the total conidia of each substrate between summer and monsoon indicate no significant difference; p > 0.05; paired t-test) species bracket leaf root rhizome trapped leaf anguillospora crassa ingold (5) (12) 36 (5) a. longissima (sacc. & p. syd.) ingold 7 (8) 176 (9) 12 192 (11) campylospora chaetocladia ranzoni (2) clavariopsis aquatica de wild. (1) (23) c. azlanii nawawi (2) (2) clavatospora tentacula sv. nilsson (2) cylindorcarpon sp. 7 2 2 35 (3) flabellospora crassa alas. 2 flagellospora curvula ingold 2 (2) 2 42 (12) f. penicillioides ingold 92 13 helicomyces louisianensis goos (2) h. roesus link (2) isthmotricladia gombakiensis nawawi (6) lateriramulosa quadriradiata k. mirua & okano 6 lunulospora curvula ingold 5 2 l. cymbiformis k. miura (3) phalangispora constricta nawawi & j. webster 1 speiropsis pedatospora tubaki 3 6 titaea clarkeae ellis & everh. 5 trinacrium subtile riess 5 3 1 tripospermum myrti (lind) s. hughes 2 26 triscelophorus acuminatus nawawi 4 (6) 4 (8) (21) t. konajensis k.r. sridhar & kaver. 2 4 (26) 3 (8) t. monosporus ingold (2) trisulcosporium acerinum h.j. huds. & b. sutton 4 varicosporium elodeae w. kegel 1 volucrispora graminea ingold, p.j. mcdougall & dann 3 total species 7 (5) 8 (3) 5 (4) 15 (13) cumulative species 7 (5) 10 (6) 10 (9) 18 (15) total conidia/g dry mass 29a (23a) 287a (24a) 33a (41a) 363a (95a) 24 k. s. karamchand and k. r. sridhar fig. 2. mean number of species (a) (n=5, mean±sd) and mean conidia (n=5) per species per gram dry mass (b) of bracket leaf, root, rhizome (drynaria quercifolia) and trapped leaf litter of sampaje region. table 3 diversity and evenness of water-borne hyphomycetes in tissues of drynaria and trapped leaves during summer and monsoon (in parenthesis) in konaje and sampaje regions tissue diversity evenness simpson shannon simpson shannon konaje bracket leaf 0.758 (0.580) 2.538 (1.726) 0.812 (0.672) 0.650 (0.615) root 0.701 (0.719) 2.110 (2.629) 0.798 (0.867) 0.703 (0.829) rhizome 0.567 (0.833) 1.299 (2.777) 0.800 (0.919) 0.819 (0.876) trapped leaf 0.388 (0.724) 1.410 (2.200) 0.427 (0.827) 0.424 (0.733) sampaje bracket leaf 0.850 (0.783) 2.620 (2.127) 0.958 (0.938) 0.933 (0.916) root 0.522 (0.620) 1.384 (1.406) 0.595 (0.891) 0.461 (0.877) rhizome 0.712 (0.551) 1.919 (1.413) 0.864 (0.717) 0.827 (0.707) trapped leaf 0.683 (0.859) 2.353 (3.099) 0.730 (0.921) 0.602 (0.838) association of water-borne fungi 25 highest in bracket leaves of drynaria in konaje and sampaje regions during summer season (tab. 3). during monsoon, the diversity was highest in rhizome of konaje, while in trapped leaf litter of sampaje. the jaccard’s similarity index revealed 35% to 53% between the substrates in konaje, while it was less similar (19-46%) in sampaje (tab. 4). live and dead parts of forest tree canopies accommodate a variety of fungi. although extensive studies have been conducted on species composition and ecological functions of water-borne fungi in woodland streams, their vertical distribution and ecological significance in riparian tree canopies is not clearly understood. these fungi are at risk due to unidirectional flow of water in streams and they overcome their extinction through colonizing the stationary substrates in stream column and along the stream banks (e.g., wood, riparian roots). similarly, tree canopy substrates (e.g., leaves, bark) and habitats (e.g., tree holes, epiphytes) serve as potential shelter for these fungi. the role of conidial fungi in canopy of douglas fir constitute a guild and serve as intermediaries of energy flow similar to aquatic hyphomycetes in stream habitats (carroll 1981). the current study demonstrated that the dead or live tissues of drynaria and trapped leaf litter in riparian trees are potential substrates for colonization of waterborne hyphomycetes. these fungi may persist in canopy during summer and perpetuate in rainy season as evident in dry leaves on the stream banks (sanders, webster 1978; sridhar, kaveriappa 1987). the live rhizome of drynaria in konaje and sampaje region yielded relatively lowest number of fungi suggests that water-borne hyphomycetes exist in rhizome as endophytes or epiphytes as suggested by gönczöl and révay (2003). recently, four water-borne hyphomycetes (dwayaangam colodena, retiarius sp., tripospermum camelopardus and t. myrti) were endophytic in black spruce (picea mariana) needles in a mixed wood forest of canada indicating their role as endophytes in tree canopies (sokolski et al. 2006). the roots of drynaria screened in our study were partially live, thus, the number of species and conidial output were fewer than those found in dead bracket leaves or trapped leaves. the overall conidial output from tissues of drynaria and trapped leaf litter although not equivalent to stream leaf litter, it is comparable to the dry leaf litter on stream banks (sridhar, bärlocher 1993) and the species richness was between 40% (konaje stream) and 75% (sampaje stream). except for rhizome, the conidial output in rest of the substrates showed significant difference between summer and monsoon in konaje, but it was not so in sampaje possibly due to cooler conditions. arborispora sp. with its characteristic multiradiate conidia was common in intact leaves, stem flow, through fall and tree snow (ando 1992). trinacrium spp. are also primarily known from rain waters, stem flow, through fall and tree holes (ando 1992; gönczöl, révay 2003, 2004). tripospermum spp. sporulated abundantly on table 4 jaccard’s similarity index (%) of bracket leaf, root, rhizome and trapped leaf of drynaria of konaje and sampaje (in parenthesis) regions root rhizome trapped leaf bracket leaf 38 (46) 35 (31) 53 (39) root 53 (39) 50 (29) rhizome 42 (19) 26 k. s. karamchand and k. r. sridhar live leaves and more prevalent in tree canopies (ando 1992; gönczöl, révay 2004). the majority of spores found in the tree canopies are multiradiate (gönczöl, révay 2006; sridhar et al. 2006). these conidia might have adapted to canopy habitats and their branches may resist total removal from the leaf or bark surface. however, in our study, fungi with sigmoid conidia were also common (e.g., anguillospora crassa and a. longissima). among the helicosporus fungi, helicomyces spp. and helicosporium sp. were rare in tree canopies. out of 37 fungi recorded in our study, up to 21 are known from the tropical tree canopies (sridhar et al. 2006; karamchand, sridhar 2008) and the rest constitute new records (e.g., clavariopsis aquatica, c. azlanii, clathrosphaerina zalewaskii, helicomyces louisianensis, magdalaenaea monogramma, speiropsis pedatospora, lateriramulosa quadriradiata, titaea clarkeae, tetracladium marchalianum, tricladium malaysianum, trisulcosporium acerinum and tumularia aquatica). sexual states of many aquatic hyphomycetes have been reported on plant detritus from terrestrial habitats in the proximity of streams (webster 1992). webster (1992) and shearer (1992) opined that wood in terrestrial habitats and emergent portions of riparian vegetation consists of teleomorphs of water-borne hyphomycetes. it is likely the perfect state of fungi might exist in tree canopies as well and disseminate their asexual propagules to streams through aerosols, raindrops and stemflow. bandoni (1981) predicted that conidia of fungi produced in tree canopies transferred directly into streams. as the bracket leaves and rachis of nest leaves of drynaria persists even after their death on the tree canopies, further observations of dead tissues may reveal perfect states of water-borne hyphomycetes. acknowledgements. authors are indebted to mangalore university for granting permission to carry out this study at the department of biosciences. one of us (ksk) acknowledges the research fellowship granted under sc/st cell, mangalore university and rajeev gandhi fellowship, university grants commission, new delhi. we thank b.s. kadamannaya for travel and field assistance. references ando k. 1992. a study of terrestrial aquatic hyphomycetes. trans. mycol. soc. japan 33: 415–425. ando k., tubaki k. 1984. some undescribed hyphomycetes in the rain drops from intact leaf-surface. trans. mycol. soc. japan 25: 21–37. bärlocher f. 1992. the ecology of aquatic hyphomycetes. springer-verlag, berlin. bärlocher f. 2006. fungal endophytes in submerged roots. (in:) b. schulz, c. boyle, t. sieber (eds). microbial root endophytes. springer-verlag, berlin and new york: 179ñ190. bandoni r.j. 1981. aquatic hyphomycetes from terrestrial litter. (in:) d.t. wicklow, g.c. carroll (eds). the fungal community its organization and role in the ecosystem, marcel dekker, new york: 693–708. belliveau m.j.-r., bärlocher f. 2005. molecular evidence confirms multiple origins of aquatic hyphomycetes. mycol. res. 109: 1407–1417. campbell j., shearer c., marvanová l. 2006. evolutionary relationships among aquatic anamorphs and teleomorphs: lemonniera, margaritispora and goniopila. mycol. res. 110: 1025–1033. carroll g.c. 1981. mycological inputs to ecosystem analysis. (in:) wicklow d.t., g.c. carroll (eds). the fungal community – its organization and role in ecosystem, marcel dekker, new york: 25–35. czeczuga b., orłowska m. 1998. hyphomycetes in rain water draining from intact trees. roczniki akademii medycznej w białymstoku 43: 66–84. gönczöl j., révay á. 2003. treehole fungal communities: aquatic, aero-aquatic and dematiaceous hyphomycetes. fungal diversity 12: 19–24. association of water-borne fungi 27 gönczöl j., révay, á. 2004. fungal spores in rainwater: stemflow, throughfall and gutter conidial assemblages. fungal diversity 16: 67–86. gönczöl j., révay á. 2006. species diversity of rainborne hyphomycete conidia from living trees. fungal diversity 22: 37–54. goos r.d. 1987. fungi with a twist: the helicosporus hyphomycetes. mycologia 79: 1–22. ingold c.t. 1942. aquatic hyphomycetes of decaying alder leaves. trans. br. mycol. soc. 25: 339–417. ingold c.t. 1975a. an illustrated guide to aquatic and waterborne hyphomycetes (fungi imperfecti). freshwater biological association scientific publication 30, united kingdom. ingold c.t. 1975b. convergent evolution in aquatic fungi: the tetraradiate spore. bot. j. linn. soc. 7: 1–25. karamchand k. s., sridhar, k. r. 2008. water-borne conidial fungi inhabiting tree holes of the west coast and western ghats of india. czech mycology 60: 63–74. kenkel n.c., booth t. 1992. multivariate analysis in fungal ecology. 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(in:) f. bärlocher (ed.). the ecology of aquatic hyphomycetes. springer-verlag, berlin: 99–117. webster j., descals e. 1979. the teleomorphs of water-borne hyphomycetes from freshwater. (in:) b. kendrick (ed.). the whole fungus. national museums of canada, ottawa: 419–447. zhao g.z., liu x.z., wu w.p. 2007. helicosporus hyphomycetes from china. fungal diversity 26: 313– 524. 2014-01-01t11:48:38+0100 polish botanical society is a remarkable species – caloplaca flavescens (lichenized fungi) – new to the polish lichen biota? lucyna śliwa and karina wilk laboratory of lichenology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, l.sliwa@botany.pl; k.wilk@botany.pl śliwa l., wilk k.: is a remarkable species – caloplaca flavescens (lichenized fungi) – new to the polish lichen biota? acta mycol. 43(2): 207–213, 2008. caloplaca flavescens is reported for the first time from poland with all known localities citation. diagnostic characters for the species are narrow, convex marginal lobes that are separated by furrows and gray crystals within the thallus cortex that are prominent in polarized light as well as lemon-shaped ascospores. species of similar appearance include c. aurantia, c. thallinicola and some members of c. saxicola group. differences between these species are briefly discussed. an up-to-date distribution map for c. flavescens in poland is presented. key words: lichens, caloplaca, new records introduction caloplaca flavescens (huds.) j.r. laundon is a member of the lobate species of the genus caloplaca and it is included in the subgenus gasparrinia tornab. defined as having lobate thallus and containing anthraquinones (giving k+ violet-red reaction) and it is included in the group c. aurantia characterized by lemon-shape ascospores. the species is distinctive due to narrow, convex marginal lobes that are separated by furrows and gray cortical crystals that are prominent in polarized light. it is calcicolous lichen occurring most often on limestone outcrops. the nomenclature and taxonomic position of the species was established by laundon (1984). clauzade and roux (1985) contributed significantly to the understanding of the species and provided its circumscription and excellent demonstrative illustrations. the species was detailed treated and widely discussed in the most recent revision of c. aurantia group in the czech republic by šoun (2005) and in the monograph of lobate species of caloplaca with a special reference to c. saxicola group in europe by gaya (2005). the taxon was noted for the first time from poland by nowak and tobolewski (1975) under the name caloplaca aurantia var. heppiana (müll. arg.) poelt, however, no data was quoted regarding collecting site of this particular variety and no acta mycologica vol. 43 (2): 207–213 2008 it is our great pleasure to dedicate this paper to professor krystyna czyżewska on the occasion of her anniversary 208 l. śliwa and k. wilk such material was found at kram herbarium. under the species name caloplaca flavescens the taxon was recognized for the first time during lichenological studies of the krakowsko-częstochowska upland by j. nowak in 1986 and of the beskid sądecki range in the polish western carpathians by l. śliwa in 1992. the species was collected from single localities and the specimens were housed in kram and kra herbaria respectively. the records, however, have never been published as they were considered by the collectors as uncertain. the determination was confirmed by c. wetmore in 1997. while visiting kram herbarium at the same time wetmore revised the material of c. aurantia group and majority of the collection was re-identified as c. flavescens. moreover, during a joint field trips with l. śliwa to different parts of southern poland he collected the species himself and the material is housed at min herbarium. based on the above collections the species was included in the checklists of the western carpathians (bielczyk 2003; bielczyk et. al. 2004) and subsequently indicated in the recent polish checklist (fałtynowicz 2003). no information concerning collecting sites were provided in any of the publications. during the recent study of calcicolous species of caloplaca in the polish western carpathians by k. wilk all material was revised and some new localities of c. flavescens were found (wilk 2008). the species appeared much easier determinable but based primarily on some anatomical characters and more frequent than it was considered before. the main purpose of this account is to make the status of c. flavescens in poland clear and to encourage lichenologists to pay special attention to this remarkable species. material and methods apart from the authors’ collections, reference materials of caloplaca aurantia (pers.) hellb., c. flavescens (huds.) j.r. laundon and c. saxicola complex from the herbaria kra, kram, and krap were investigated. for light microscopy, free-hand sections were made with a razor blade and mounted in water. tissue and ascospore measurements were made in water. measurements are rounded to the nearest 0.5 microns. granulation of tissues was observed in polarized light. the solubility of granules or/and crystals was tested with ca 25% koh (k) and 65% nitric acid (n); these last two reagent tests were always carried out on separate cross sections. spot test reactions of thalli, apothecial margins and discs were made with the same reagents. description of the species caloplaca flavescens (huds.) j.r. laundon, lichenologist 16: 53. (1984). figs 1a, 2a, 3a basionym: lichen flavescens huds., fl. anglica: 445. 1762. type: hist. musc.: 136, tab. 18, fig. 18a (bm – lectotype, not seen). selected synonyms: caloplaca heppiana (müll. arg.) zahlbr.; amphiloma heppianum müll. arg. caloplaca aurantia var. heppiana (müll. arg.) poelt [for details see laundon 1984]. is a remarkable species 209 thallus placodioid, forming closely adpressed rosettes up to 3 cm in diameter; rosettes of thalli single or grouped, yellowish orange or deep orange, sometimes blanched in the central part of thallus, especially if growing in the deep shadow, epruinose or faintly pruinose; lobes at the margin of thallus 5–10 mm long and 0.3– 0.5 mm wide, deeply and dense incised, narrow and convex, separated by furrows, lobe-ends slightly broader and indented; center of thallus with uneven, irregular and often convex areoles; cortex thick, prosoplectenchymatic with thick layer of coarse, gray crystals (prominent in polarized light, insoluble in k and slowly soluble in n); algae layer discontinuous, algae cells clustered into groups; medulla prosoplectenchymatic, without crystals. apothecia up to 1.2 mm diam., mainly confined to the centre of the thallus, usually abundant, dispersed or crowded, regular in shape or angular and flexuose due to compression; disc flat to convex, deep orange to orange brownish, epruinose; true exciple thin, prominent or level with the disc, smooth, paler than disc; thalline exciple soon excluded, sometimes visible at the base of the apothecium. amphithecium lowered, algae abundant, cortex ±thick, paraplectenchymatie; parathecium well developed, distinct, prosoplectenchymatic; epihymenium granular, golden brown; hymenium hyaline, 80–115 µm high; hypothecium hyaline, prosoplectenchymatic, with numerous oil droplets. paraphyses thick, up to 2.5 µm wide, simple or faintly branched at the top, not expanded, or slightly expanded apically. spores broad, lemon-shaped, 8–15 × 6–11.5 µm, septum (2–)3–4 µm, 8 per ascus. pycnidia often abundant, orange, immersed; conidia simple, straight, colorless, 3.5–5.5(–7.5) × 1 µm. spot test reactions. thallus and apothecia k+ violet-red, n–; medulla k–, n–; epihymenium k+ violet-red. discussion. caloplaca flavescens can be rather easily distinguished by long, narrow and convex marginal lobes and spores that are lemon-shaped. moreover, the thallus cortex of the species is obscured by wide layer of grey crystals (fig. 2a) that are prominent in polarized light (fig. 3a). due to these character it is rather unlikely to be mistaken with any other caloplaca member. the species is mostly related to c. aurantia (fig. 1b). the latter species, however, lacks crystals in the thallus cortex (figs 2b, 3b). the thalline lobes of c. aurantia are flat and distinctly broader at the ends and closely attached one to the other. moreover, the species is lighter and often zonated in colour. both species are widely discussed by šoun (2005) and gaya (2005). representatives of c. saxicola complex have much smaller thallus with considerable shorter lobes that are often pruinose or scabrid. the members of the group have also ellipsoid spores. most of species of the group differ also in habitat preferences. another similar taxon is c. thallinicola (wedd.) du rietz. it differs from c. flavescens by very elongated lobes, deeper furrows between lobes and cortical crystals clustered into spherical, separated groups (see clauzade & roux 1985). most of all, however, the species differs in ecology. it occupies maritime acid and calcareous rocks. remarks. as a result of c. flavescens delimitation a distribution of c. aurantia in poland is in need of urgent revision. the species is most likely more rare than indicated in bibliographic sources. is a remarkable species 211 well as in scandinavian countries (norway and sweden: santesson et al. 2004). it was also reported from near and middle east (israel: kondratyuk et al. 1996b; turkey: john 1996) and northern africa (egypt: seaward and sipman 2006; morocco: egea 1996; tunisia: seaward 1996). according to šoun (2005) it was also recorded in canary islands, azores and european part of russia. specimens examined. poland. carpathians: beskid sądecki mts., pasmo radziejowej, szlachtowa, alt. 550 m, fence made of calcareous sandstones, 2 may 1991, l. śliwa s.n. (kra); pieniny mts., wąwóz homole canyon, 49°23’54”n/20°33’20”e, 30 aug. 2000, c. wetmore 85268 (min); pieniny małe mts.: “biała woda”, calcareous rock on right bank of a stream, near old sheepfold, 1 june 1999, j. kiszka s.n. (krap); dziobakowe skały rocks, se of jaworki village, alt. 738 m, 49°23’21’’n/20°34’05’’e, large limstone outcrops in a mixed forest, 4 june 2005, k. wilk 3429, 3435 (kram); near jaworki village, limstone outcrops along bike trail, alt. 580 m, 49°24’20’’n/20°32’30’’e, 3 june 2005, k. wilk 3423 (kram); podhale foothills, skalice nowotarskie, n slope of kramnica mt., calcareous rock, alt. 640 m, 24 june 1968. j. kiszka s.n. (krap); tatry zachodnie mts.: dolina kościeliska valley, 3 km sw of zakopane, 49°14’30”n/19°51’46”e, 9 may 1997, c. wetmore 77318 (min); dolina kościeliska valley, near wyżnia pisana polana glade, vertical calcareous rock, alt. ca 1100 m, 13 july 2004, k. wilk 2129; wąwóz kraków canyon, vartical calcareous rock, alt. ca 1100 m, 15 july 2004, k. wilk 2147, 2149 (kram). wyżyna krakowsko-częstochowska upland: limestone quarry, 12 km nw of krakow, along a road to olkusz, 50°09’12”n/19°47’06”e, 6 may 1997, c. wetmore (min); dolina będkowska valley, alt. 380 m, vertical limestone rocks, 1 may 1956, j. nowak (kram-l 18770); dolina kluczwody valley, at the base of vertical limestone rocks, 19 dec. 1992, j. nowak (kram-l 31608), zamkowa rock, alt. 320 m, in a bottom of the valley, at the base of vertical limestone rock, 15 june 1994, j. nowak (kram-l 28036); dolina kobylańska valley, kula rock, alt. 350 m, vertical limestone rock, in a sunny place, 2 may 1995, j. nowak (kram-l 40213); nielepice village near rudawa, 6 km ees of krzeszowice town, n-facing, open slope of chełm hill, alt. 280 m, on vertical or overhanged limestone outcrops, 30 sept. 1986, j. nowak (kram-l 30454). additional specimens examined. bulgaria. czarnatica. bielocerkowski rid, biala cerkwa, alt. 1300 m, calcareous rocks, 29 sept. 1975, j. nowak (kram-l 34966, 34968). slovakia. spisz, drevenik near spissky hrad, alt. 600 m, calcareous rocks, 3 july 1993, u. bielczyk (kram-l 23972). acknowledgements. sincere thanks are due to prof. dr. clifford m. wetmore (university of minnesota, usa) for confirmation or revision of some polish collections. we are grateful to the anonymous reviewer for valuable suggestions on the manuscript. the study was supported by the ministry of science and higher education, grant no. n303 294334. references bielczyk u. 2003. the lichens and allied fungi of the polish western carpathians. 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1996. catalogue of lichenized and lichenicolous fungi of marocco. bocconea 6: 19-114. fałtynowicz w. 2003. the lichens, lichenicolous and allied fungi of poland. an annotated checklist. w. szafer institute of botany, polish academy of sciences, kraków, 435 pp. gaya e. 2005. revisió morfològica i molecular dels tàxons lobulats del gènere caloplaca (teloschistaceae, liquens), amb especial èmfasi en el grup de c. saxicola. doctoral thesis, university of barcelona. 419 pp. available at http://www.tesisenxarxa.net/, date of exploration: 20 october 2008. hafellner j., türk r. 2001. die lichenisierten pilze �sterreichs – eine checkliste der bisher nachgewiese-ürk r. 2001. die lichenisierten pilze �sterreichs – eine checkliste der bisher nachgewiese-rk r. 2001. die lichenisierten pilze �sterreichs – eine checkliste der bisher nachgewiese-�sterreichs – eine checkliste der bisher nachgewiese-sterreichs – eine checkliste der bisher nachgewiesenen arten mit verbreitungsangaben. stapfia 76: 3-173. john v. 1996. preliminary catalogue of lichenized and lichenicolous fungi of mediterranean turkey. bocconea 6: 173-216. kondratyuk s.ya., navrotskaya i., khodosovtsev a.ye., solonina o. 1996a. checklist of ukrainian lichens. bocconea 6: 217-294. kondratyuk s.ya., zelenko s.d., wasser s.p., nevo e. 1996b. the first checklist of lichen-forming and lichenicolous fungi of israel. m.h. kholodny institute of botany, national academy of sciences of ukraine & international centre for cryptogamic plants and fungi, kiev, haifa, 136 pp. kondratyuk s.ya., popova l.p., lackovičová a., pišút i. 2003. a catalogue of eastern carpathian lichens. m.h. kholodny institute of botany, national academy of sciences of ukraine & institute of botany, slovak academy of sciences, kiev, bratislava, 263 pp. laundon j.r. 1984. studies in the nomenclature of british lichens i. lichenologist 16 (1): 53-57. llimona x., hladun n.l. 2001. checklist of the lichens and lichenicolous fungi of the iberian peninsula and balearic islands. bocconea 14: 5-581. mayrhofer h., denchev c.m., stoykov d.y., nikolova s.o. 2005. catalogue of the lichenized and lichenicolous fungi in bulgaria. mycologia balcanica 2(1): 3-61. nimis p.l. 1993. the lichens of italy. an annotated catalogue. museo regionale di scienze naturali, monografie 12, torino, 897 pp. nimis p.l., martellos s. 2003. a second checklist of the lichens of italy with a thesaurus of synonyms. museo regionale di scienze naturali, monografie 4, aosta, 4192 pp. nowak j., tobolewski z. 1975. porosty polskie. opisy i klucze do oznaczania porostów w polsce dotychczas stwierdzonych lub prawdopodobnych. państwowe wydawnictwo naukowe, warszawa-kraków, 1177 pp. pišút i., lackovičová a., lisická e. 1996. a second checklist and bibliography of slovak lichens. biologia, bratislavia, 51/supplement 3: 1-79. santesson r., moberg r., nordin a., tønsberg t., vitikainen o. 2004. lichen–forming and lichenicolous fungi of fennoscandia. museum of evolution, uppsala university, uppsala, sweden, 359 pp. seaward m.r.d. 1996. checklist of tunisian lichens. bocconea 6: 115-148. seaward m.r.d., sipman h. 2006. an updated checklist of lichenized and lichenicolous fungi for egypt. willdenowia 36: 537-555. šoun j. 2005. revize skupiny caloplaca aurantia v ceské republice mgr. thesis, faculty of biological sciences, university of south bohemia, ceské budejovice, czech republic, 46 p., + 2 p. suppl., available at http://botanika.bf.jcu.cz/thesis/pdf/soun2005.pdf, date of exploration: 20 october 2008. wilk k. 2008. kalcyfilne gatunki rodzaju caloplaca w polskich karpatach zachodnich. doctoral thesis, w. szafer institute of botany, polish academy of sciences, kraków, 158 pp. wirth v. 1995. die flechten baden–württembergs, teil 1 & 2. eugen ulmer gmbh & co., stuttgart, 1006 pp. is a remarkable species 213 czy caloplaca flavescens jest gatunkiem nowym dla bioty porostów polski? s t r e s z c z e n i e caloplaca flavescens (huds.) j.r. laundon (fig. 1a) reprezentuje grupę caloplaca ujętych w podrodzaju gasparrinia, wytwarzających zewnętrzną łatkowatą plechę i zawierających antrachinony. jest to porost kalcyfilny, występujący na wychodniach skał wapiennych. w polsce gatunek ten był wyróżniony dawno temu i choć w literaturze powoływano się na okazy zielnikowe to nigdy nie został właściwie opublikowany. tutaj gatunek c. flavescens jest po raz pierwszy odnotowany z polski i udokumentowany materiałami zielnikowymi. ponadto podano jego charakterystykę i porównano z gatunkami, z którymi może być mylony – z c. aurantia (fig. 1b), grupą c. saxicola oraz c. thallinicola. caloplaca flavescens charakteryzuje się długimi, wąskimi i wypukłym łatkami, które są oddzielone głębokimi szczelinami. zarodniki c. flavescens mają cytrynowaty kształt a kora plechy wypełniona jest szeroką warstwą szarych kryształów świecących w świetle spolaryzowanym (figs 2a, 3a). najbliżej gatunek ten jest spokrewniony z c. aurantia. łatki plechy c. aurantia są płaskie, wyraźnie szersze na końcach i ściśle do siebie przylegające. cechą diagnostyczną dla tego gatunku jest także brak wyżej opisanych kryształów w korze (fig. 2b, 3b). przedstawiciele grupy c. saxicola mają mniejszą plechę, znacznie krótsze, często przyprószone łatki oraz posiadają elipsoidalne zarodniki. innym podobnym taksonem jest c. thallinicola. gatunek ten różni się od c. flavescens bardzo wydłużonymi łatkami oraz głębokimi szczelinami pomiędzy nimi, jak również kryształami w korze plechy, które łączą się w kuliste skupienia. ponadto gatunek ten zajmuje odmienne siedlisko, mianowicie występuje na skałach morskich, zarówno kwaśnych jak i zawierających węglan wapnia. w pracy omówiono także siedliska zajmowane przez c. flavescens, znane rozmieszczenie tego gatunku w polsce (fig. 4) oraz jego ogólny zasięg występowania. 2014-01-01t11:48:17+0100 polish botanical society 246 m. mańka the warsaw agricultural university (sggw) in 1952, and started her work in central office for fruit and vegetable industry in warsaw. she obtained master of science in plant pathology at the warsaw agricultural university in 1958, starting her work in the division of institute of plant protection in reguły, near warsaw. soon afterwards (in 1959) prof. pokacka went to the department of botany and plant pathology (purdue university in lafayette, indiana, usa) where in 1962 she obtained her d.sc. degree based on a thesis: “studies on soil microflora and biological control of damping-off of apple”. after coming back in 1962 dr pokacka continued her work in reguły near warsaw, dealing mainly with root diseases of vegetables. the following years were filled with work in gorzów wielkopolski: first in the department of forage plants of the institute of soil science and plant cultivation (iung, 1967-1970), and than in the institute of plant protection branch in gorzów wielkopolski (1970-1972). that was the beginning of her work on cereal diseases, continued till her retirement in 1994. the research was focused on: snow mould (fusarium nivale [= microdochium nivale]), leaf and stem base diseases of wheat, rye and triticale – with a special reference to their occurrence and diagnostics, resistance of wheat and rye cultivars to rusts and powdery mildew under high nitrogen fertilization. from 1972 on prof. pokacka was head of the department of mycology in the institute of plant protection in poznań and her new task became supervising research on puccinia triticina and erysiphe graminis (= blumeria graminis) races on wheat, followed by the problem of erysiphe graminis f. sp. tritici resistance to triazole fungicides. than her research interests shifted to physiological races of barley pathogen, rynchosporium secalis. examining new fungicides against cereal diseases, started in late 1970ties, resulted in the first instructions for complex cereals protection in poland. after gaining the degree of doctor habilitatus in 1976 (habilitation dissertation “grzyby drożdżoidalne występujące na ziarniakach pszenicy” – “yeast fungi occurring on kernels of winter wheat”), professor pokacka further investigated cereal diseases: sharp eyespot (rhizoctoniosis) of cereal stem base, and pythium vanterpoolii – a new pathogen of cereals and grasses in belgium, septoriosis of wheat (and later triticale) ears and leaves caused by septoria (= stagonospora) nodorum, and tan leaf spot of wheat and triticale caused by drechslera tritici-repentis. results of research work carried out by prof. pokacka and her team on the occurrence and chemical control of cereal diseases supplied genuine basic information for initiating, for the first time in poland, modern programmes of cereal disease control. a remarkable part of prof. pokacka scientific output consists of review articles concerning new science/research achievements, such as mechanisms of fungicide activity, side effects of fungicides and etiology of plant diseases. the scientific output of professor zofia pokacka consists of 155 works, including 69 research papers and 86 scientific reports and articles. she promoted five doctors of agriculture and reviewed eight other ph.d. theses, one habilitation dissertation and a number of applications for research grants. in 1976 doctor habilitatus zofia pokacka was appointed associated professor and in 1989 the council of the state granted her the title of full professor. during her leadership of the department of mycology in the institute of plant protection (1972-1992) prof. pokacka had also represented poland in an european professor zofia maciejowska-pokacka 247 and mediterranean plant protection organization (eppo) working group for methods of pesticide efficacy estimation in protection of agricultural and horticultural plants. she was also member of scientific councils in institute of plant protection (1981-1992), research centre for cultivar testing (centralny ośrodek badania odmian roślin uprawnych; 1978-1983), institute of soil science and plant cultivation (instytut uprawy, nawożenia i gleboznawstwa; 1987-1992). in 1987-1992 she was a member of the committee of plant protection of the polish academy of sciences, and in 1997-1999 – a member of the agricultural section of the state committee for scientific research. professor pokacka was a member of siitr (professional organization of agricultural engineers and technicians), polish botanical society and the polish phytopathological society (since 1972). she was an active member of the poznań division with keen interest in its work and development – she was a member of the division board (1985-1996) and division board of controllers (1976-1978). after being a member of the entire society board of controllers (1979-1981), in 1982-1984 she was a member of the society board. the resolution of the general assembly of members of the polish phytopathological society in 1999 in poznań granted her the title of honorary member of the society. as a member of the polish phytopathological society, she was also member of the international society for plant pathology and european foundation for plant pathology. in 2003 volume 28 of phytopathologia polonica was dedicated to professor zofia pokacka on the occasion of the 45th anniversary of her scientific work (mańka 2003a and 2003b). professor zofia pokacka’s editorial work was long lasting and intense. from 1986 till 2001 prof. pokacka was a division editor and editorial committee member of journal of plant protection/prace naukowe ior, progress in plant protection research/postępy w ochronie, and from 1991 on she has been a member of the editorial committee of phytopathologia polonica (from 2009 – phytopathologia; scientific journal of the polish phytopathological society). her contribution to editorial work in plant pathology, in the form of reviewing papers and editing scientific periodicals, was enormous. among the awards for research and extension work given to professor zofia pokacka there were: state award iiº in 1980 and the awards of siitr and of minister of agriculture, the golden cross of merit (złoty krzyż zasługi; 1977) and decoration “meritorious to agriculture” (odznaka honorowa “zasłużony dla rolnictwa”; 1983), medal 40-years of polish people’s republic (medal 40-lecia polski ludowej; 1984), and commemorative medal of the institute of plant protection issued on the 50th anniversary of its establishing. professor pokacka will be long remembered not only for her professional accomplishments, but also for her friendliness and modest nature. she was a person of great heart, always ready to help younger colleagues in research and publication work. her virtues are a great heritage to her friends and co-workers. references mańka m. 2003a. forty five years of scientific work of professor zofia maciejowska-pokacka d.sc. (agr.). phytopathol. pol. 28: 9–12. mańka m. 2003b. list of publications by professor zofia maciejowska-pokacka d.sc. (agr.). phytopathol. pol. 28: 13–20. 248 m. mańka bibliography of publications by zofia maciejowska-pokacka (1959-1995) karpińska-maciejowska z. 1959. z badań nad patogenicznością grzyba phoma lingam (tode) desm. powodującego suchą zgniliznę roślin krzyżowych. biuletyn instytutu ochrony roślin 6: 117-131. maciejowska z., williams e.b. 1959. the relation of certain fungi to root and collar rot of apple. indiana academy of sci. proc. 69: 107. maciejowska z., williams e.b. 1960. the isolation and indentification of fungi and their relation to root rot of apple. indiana academy of sci. proc. 70: 52-53. świrska s., łacicowa b., maciejowska z. 1960. obserwacje nad żywotnością i mykoflorą 4 gatunków nasion niezaprawianych, przechowywanych w różnych warunkach wilgotności powietrza. biuletyn instytutu hodowli i aklimatyzacji roślin 4: 47-56. maciejowska z. 1963. z pobytu na praktyce naukowej na uniwersytecie purdue w stanie indiana ameryki północnej. postępy nauk rolniczych 2 (80): 153-156. maciejowska z., williams e.b. 1963. studies on morphological forms of staphylotrichum coccosporum. mycologia lv (2): 221-225. maciejowska z., williams e.b. 1963. studies on a multiloculate species of preussia. mycologia lv (3): 300-308. maciejowska z., williams e.b. 1963. the effect of cellulose additions and moisture level on mycoflora of soil. canad. j. microbiol. 9: 555-561. maciejowska z. 1964. choroby korzeni roślin na tle ekologii grzybów glebowych. biuletyn instytutu ochrony roślin 26: 177-203. maciejowska z. 1964. wpływ zabiegów ochronnych i mikroflory glebowej na występowanie niektórych chorób korzeni warzyw. prace naukowe instytutu ochrony roślin 8 (2): 153-162. maciejowska z. 1964. przyczyny chorób korzeni roślin kapustnych na glebie torfowej w regułach. biuletyn instytutu ochrony roślin 28: 43-47. maciejowska z. 1965. grzyby z rodzaju trichosporon wyizolowane z gleby torfowej. prace naukowe instytutu ochrony roślin 6 (1): 61-91. maciejowska z. 1965. zgorzel siewek warzyw i jej zwalczanie. hasło ogrodniczo-rolnicze 12: 384-385. maciejowska z. 1966. obserwacje nad morfologią geotrichum sp. z korzenia kapusty. acta mycol. polon. 2: 175-181. maciejowska z. 1966. zwalczamy mączniaka rzekomego sałaty w szklarniach i inspektach. hasło ogrodniczo-rolnicze 2: 40-41. maciejowska z. 1967. wyniki badań nad zwalczaniem chorób korzeni kapusty na glebie torfowej w regułach. prace naukowe instytutu ochrony roślin 9 (1): 41-49. maciejowska z. 1967. znaczenie integrowanej metody walki z chorobami w ochronie roślin. nowe rolnictwo 11: 16-17. maciejowska z. 1967. integrowana metoda walki z chorobami roślin wywoływanymi przez grzyby. biuletyn instytutu ochrony roślin 36: 249-256. maciejowska z. 1967. z badań nad mikoflorą gleby torfowej i jej wpływem na zdrowotność korzeni niektórych roślin kapustnych. prace naukowe instytutu ochrony roślin 9 (2): 117-144. maciejowska z., bogusławski w. 1968. badania nad patogenicznością grzyba rhizoctonia solani kűhn w stosunku do fasoli szparagowej. prace naukowe instytutu ochrony roślin 10 (1): 101-112. maciejowska-pokacka z. 1969. z badań nad mikoflorą chorych korzeni fasoli szparagowej. prace naukowe instytutu ochrony roślin 10 (2): 135-143. maciejowska z. 1970. zgorzel siewek roślin kapustnych i jej zwalczanie. prace naukowe instytutu ochrony roślin 12 (1): 205-226. maciejowska-pokacka z. 1971. wyniki jednorocznych badań nad wpływem różnych gleb na mikoflorę przy uprawie kupkówki. acta mycol. polon. 7 (1): 31-40. maciejowska-pokacka z. 1971. reakcja mikoflory glebowej i innych drobnoustrojów na różne poziomy nawożenia azotem i nawadniania przy uprawie kupkówki. acta mycol. polon. 7 (1): 41-57. maciejowska-pokacka z. 1971. wstępne badania nad odpornością odmian żyta na pleśń śniegową. biuletyn instytutu ochrony roślin 50: 195-205. maciejowska-pokacka z. 1971. odporność zbóż na choroby powodowane przez grzyby rdzawnikowe. in: wybrane zagadnienia z zakresu ochrony roślin. wykłady na kursie iii stopnia dla pracowników pskior. skrypt. p. 55-59. professor zofia maciejowska-pokacka 249 pokacka z., wojtaszek d. 1972. uwagi o przydatności układowego preparatu calixin do zwalczania mączniaka właściwego erysiphe graminis d.c. w intensywnej uprawie pszenicy ozimej. biuletyn instytutu ochrony roślin 54: 75-81. pokacka z. 1972. co warto wiedzieć o rdzy źdźłowej na pszenicy. chłopska droga 67: 10. pokacka z. 1972. porażenie przez mączniak właściwy pszenicy ozimej i jej plonowanie w warunkach wysokiego nawożenia azotowego. ochrona roślin 7: 4-5. pokacka z., cieślak s. 1972. brunatnienie plew pszenicy. ochrona roślin 10: 5-6. pokacka z. 1973. odporność zrejonizowanych odmian pszenicy ozimej i żyta na niektóre choroby grzybowe. ochrona roślin 6: 3. pokacka z. 1973. kilka uwag o rynchosporiozie zbóż wywoływanej przez rhynchosporium secalis (oud.) j. j. davis. ochrona roślin 7: 5-6. pokacka z. 1973. pleśń śniegowa. audycja radiowa. chłopska droga 58: 14. pokacka z. 1973. czernienie kłosów zbóż. ochrona roślin 9: 3-4. gołębiowska z., pokacka z. 1973. kompleks czynników wpływających na ograniczenie najważniejszych chorób i szkodników zbóż. międzynarodowe czasopismo rolnicze 4: 26-31. pokacka z., błońska-pawlak a. 1973. wpływ wysokiego nawożenia azotowego na odporność polową odmian pszenicy ozimej na mączniaka właściwego (erysiphe graminis d.c.) i rdzę brunatną (puccinia triticina erikss.). biuletyn instytutu ochrony roślin 56: 153-179. pokacka z. 1974. fuzarioza żyta w 1974 r. i jej zwalczanie. ochrona roślin 9: 3. pokacka z., 1975: sporysz zbóż i traw, claviceps purpurea (fr.) tul. (stadium niedoskonałe sphacelia segetum lev.). ochrona roślin 6: 6-8. pokacka z. 1975. znaczenie hodowli odpornościowej w rolnictwie i w zwalczaniu chorób roślin (zboża). in: wybrane zagadnienia z zakresu ochrony roślin. wykłady na kursie iii stopnia dla pracowników pskior. skrypt. p. 51-55. pokacka z. 1975. najnowsze środki grzybobójcze stosowane w rolnictwie (zboża). in: wybrane zagadnienia z zakresu ochrony roślin. wykłady na kursie iii stopnia dla pracowników pskior. skrypt. p. 69-79. pokacka z., błońska-pawlak a. 1975. uwagi o polowej odporności odmian pszenicy ozimej na mączniaka właściwego erysiphe graminis d.c. f.sp. tritici. prace naukowe instytutu ochrony roślin 17 (1): 123131. pokacka z. 1975. ocena odporności polowej odmian żyta na rdzę brunatną puccinia dispersa erikss. i p. graminis erikss. et henn. f.sp. secalis w warunkach wysokiego nawożenia azotowego w latach 19701974. prace naukowe instytutu ochrony roślin 17/1/: 105-121. pokacka z., wojtaszek d. 1976. ryzoktonioza podstawy źdźbła pszenicy (rhizoctonia solani kűhn). ochrona roślin 6: 6-7. maciejowska-pokacka z. 1976. grzyby drożdżoidalne występujące na ziarniakach pszenicy. i. studium taksonomiczne szczepów candida albicans (robin) berkhout i c. tropicalis (cast.) berkhout. acta mycol. polon. 12 (1): 113-122. maciejowska-pokacka z. 1976/1977. grzyby drożdżoidalne występujące na ziarniakach pszenicy. ii. studium taksonomiczne szczepów cryptococcus laurentii (kuff.) skinner var. magnus lodder et kreger-van rij i c. albidus (saito) skinner var. albidus. acta mycol. polon. 12 (2): 195-202. maciejowska-pokacka z. 1976/1977. grzyby drożdżoidalne występujące na ziarniakach pszenicy. iii. studium taksonomiczne szczepów sporobolomyces roseus kluyver et van niel i s. pararoseus olson et hammer. acta mycol. polon. 12 (2): 203-210. maciejowska-pokacka z. 1977. grzyby drożdżoidalne występujące na ziarniakach pszenicy. iv. studium taksonomiczne szczepów cryptococcus laurentii (kuff.) skinner var. laurentii i var. flavescens (saito) lodder et kreger-van rij. acta mycol. polon. 13 (1): 3-9. pokacka z., wojtaszek d. 1977. z badań nad patogenicznością rhizoctonia solani kűhn na pszenicy i życie. materiały 17 sesji instytutu ochrony roślin. poznań. p. 181-192. pokacka z. 1977. nowe kierunki w ochronie zbóż przed chorobami. ochrona roślin 5: 3-5. pokacka z., głęboczyk b. 1978. stan badań w polsce nad nowymi fungicydami do ochrony zbóż. nowe rolnictwo 1: 16-17. pokacka z., bojarski s..1978. kompleksowa ochrona zbóż przed chorobami. instrukcja wdrożeniowa. instytut ochrony roślin. poznań. 35 pp. maciejowska-pokacka z., parmentier g. 1978. first record of a pythium pathogenic to cereals and grasses in belgium, and its identification as pythium vanterpoolii. parasitica 34 (1): 3-10. 250 m. mańka pokacka z., grala b. 1978. próba oceny metodami statystycznymi podatności rodów hodowlanych żyta na rdzę brunatną (puccinia dispersa eriksson) i mączniaka prawdziwego (erysiphe graminis d.c.). materiały 18 sesji instytutu ochrony roślin. poznań. p. 137-150. pokacka z., jańczak c. 1978. ochrona zbóż niezbędna. plon 15: 5. pokacka z. 1979. chemiczna ochrona zbóż przed chorobami w okresie wegetacji i jej wpływ na podniesienie plonów ziarna. biuletyn informacyjny zppgr. poznań 4: 14-21. pokacka z. 1979. plamistości liści pszenicy powodowane przez grzyby z rodzaju septoria sacc. cz. i. występowanie i cechy morfologiczne gatunków s. nodorum berk., s. tritici rob. ex desm. i s. avenae frank f. triticea t. johnson. ochrona roślin 5: 6-8. maciejowska-pokacka z. 1978. yeast fungi in the phyllosphere and on seeds of wheat. congress of plant pathology, műnchen, 16-23 aug. 5. iii. p. 103. pokacka z. 1979. uwagi o doświadczeniach wdrożeniowych przeprowadzonych w 1978 r. nad chemiczną ochroną pszenicy ozimej przed kompleksem chorób. ochrona roślin 5: 19-21. pokacka z. 1979. plamistości liści pszenicy powodowane przez grzyby z rodzaju septoria sacc. ii. objawy chorobowe. ochrona roślin 7: 7-10. pokacka z. 1979. plamistości liści pszenicy powodowane przez grzyby z rodzaju septoria sacc. iii. zwalczanie chemiczne i odporność odmian. ochrona roślin 7: 10-12. maciejowska-pokacka z. 1979. grzyby drożdżoidalne występujące na ziarniakach pszenicy. studium taksonomiczne szczepu bullera alba (hanna) derx. acta mycol. polon. 15 (2): 295-302. pokacka z. 1980. chemiczna ochrona pszenicy ozimej przed chorobami w doświadczeniach wdrożeniowych w 1979 r. ochrona roślin 5: 13-14. pokacka z. 1980. fuzarioza podstawy źdźbeł żyta. ochrona roślin 8: 3-5. pokacka z. 1981. choroby zbóż i ich zwalczanie. 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(in:) poradnik nawożenia i ochrony roślin agrochem 1982. warszawa. p. 317-321. pokacka z. 1982. uboczne efekty stosowania fungicydów do zwalczania chorób występujących w uprawach zbóż. prace naukowe instytutu ochrony roślin 24 (1): 5-14. pokacka z. 1983. potrzeba chemicznej ochrony zbóż przed chorobami. rolnictwo. informacje zalecenia. wopr lubań stare pole. 42 (2): 26-30. pokacka z. 1983. patogeny nasion zbóż i ich rozpoznanie. ochrona roślin 7: 4-7. pokacka z. 1983. chemiczna ochrona pszenicy ozimej przed chorobami. (in:) technologia produkcji pszenicy ozimej. materiały szkoleniowe. skrypt. instytut uprawy, nawożenia i gleboznawstwa, puławy. p. 57-67. pokacka z. 1983. mechanizmy działania fungicydów stosowanych do ochrony zbóż przed chorobami. prace naukowe instytutu ochrony roślin 25: 117-133. pokacka z. 1983. pielęgnacja i ochrona roślin. 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(stadium workowe pyrenophora teres drechs.). ochrona roślin 2/3: 3-4. maciejowska-pokacka z., jańczak c. 1985. trends in the control of cereal diseases in poland. schaderreger in der getreideproduktion. 4th international symposium. halle-wittenberg, 27-30 nov. 1984, p. 194-195. jańczak c., pokacka z. 1985. ocena wyników zwalczania chorób grzybowych w pszenicy ozimej w 1984 roku. ochrona roślin 9: 3-4. maciejowska-pokacka z. 1986. studium taksonomiczne szczepu trichosporon beigelii (kűch. et. rabenh.) vuillemin z gleby torfowej. acta mycol. polon. 18 (2): 289-296. pokacka z. 1986. ochrona roślin. (in:) kompleksowa technologia produkcji jęczmienia jarego (wyd. ii poprawione i uzupełnione). instrukcja upowszechnieniowa. instytut uprawy, nawożenia i gleboznawstwa, puławy. p. 31-33. pokacka z. 1986. ochrona roślin. 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(in:) m. mańka (ed.). environmental biotic factors in integrated plant disease control. proceedings of 3rd conference of european foundation for plant pathology, poznań, poland, september 5-9, 1994. the polish phytopathological society: p. 517-521. małgorzata mańka 2014-01-01t23:51:55+0100 polish botanical society isolation of bacteria from ectomycorrhizae of tuber aestivum vittad. milan gryndler and hana hršelová institute of microbiology ascr, vídeňská 1083, cz-14220, prague 4, gryndler@biomed.cas.cz gryndler m., hršelová h.: isolation of bacteria from ectomycorrhizae of tuber aestivum vittad. acta mycol. 47 (2): 155–160, 2012. fifteen different cultivation media were used to isolate bacteria with the idea to obtain taxa specifically associated with ectomycorrhizae of tuber aestivum. ectomycorrhizae were collected at the sampling points previously analyzed for bacterial molecular diversity. we isolated 183 bacterial strains and identified them on the basis of the partial sequence of 16s rdna. out of these isolates, only 4 corresponded to operational taxonomic units significantly associated with t. aestivum ectomycorrhizae in previous molecular study. preliminary study of the effect of 12 selected isolates on growth of t. aestivum mycelium showed no stimulation and one isolate induced the damage of hyphae. different isolation strategy has to be developed to increase the probability of cultivation of potentially important components of t. aestivum mycorrhizosphere. key words: culture, summer truffle, pseudonocardineae, streptomyces, rhizobiales introduction for centuries, truffles have been highly appreciated as a special culinary ingredient, being highly priced on the markets. although some of the truffle species have successfully been produced in plantations, attempts to culture other species have repeatedly failed and they can only be collected from their primary habitats. life strategy of different truffle species is understood insufficiently and mainly the interactions between truffles and mycorrhizaeand mycelium-associated microflora merits attention. truffles are ectomycorrhizal ascomycota deriving significant portion, if not all, of their carbon nutrition from a host tree via specialized communication organs, ectomycorrhizae. the soil space directly affected by ectomycorrhizae is called mycorrhizosphere (linderman 1988) and is characterized by high biological activity (chalot, brun 1998). presence of some bacteria may enhance the formation of ectomycorrhizal structures on the roots and was shown to change gene expression of the ectomycorrhizal fungal hyphae (deveau et al. 2007). some of these microbes acta mycologica vol. 47 (2): 155–160 2012 156 m. gryndler and h. hršelová referred to as „mycorrhization helper bacteria“ (garbaye 1994) have received a special attention because they can stimulate mycorrhizal colonization of the host roots by the truffles with economic consequences of the few available studies directed to microbial associates of truffles, some have focused on bacteria (citterio et al. 2001; bedini et al. 1999; sbrana et al. 2002) and yeasts (zacchi et al. 2003) associated with the ascocarp or ectomycorrhizae. the information on the microflora of the soil inhabited by truffles is also infrequent and has been published for yeasts by zacchi et al. (2003) and, for saprotrophic fungi by luppi-mosca (1973, in napoli et al. 2008) and for bacteria by sbrana et al. (2002). the most striking result of these studies is a strong association of yeasts cryptococcus spp. with ectomycorrhizae of the truffle ground reported by zacchi et al. (2003). some novel information is gradually becoming available using cultivation-independent molecular methods such as high-throughput pcr and dna sequencing. these approaches allowed insights into mycorrhizal community of the productive spots of t. magnatum – showing that the mycorrhizae of this fungus are generally rare and most roots of the host trees are occupied by other mycorrhizal fungi (murat et al 2005). molecular analysis of bacterial and fungal communitites associated with the productive spots of t. magnatum failed to identify any specific fungi associated with this truffle, whereas they suggested a bacterium moraxella osloensis (a gamma-proteobacterium) to be preferentially associated with the productive spots of this truffle (mello et al. 2010). our previous study (gryndler et al. 2012) revealed a specific association of some bacteria, including four genera of actinobacterial suborder pseudonocardineae, with ectomycorrhizae of tuber aestivum vittad. in this study, we aimed at isolation of bacteria from the communities associating to the t. aestivum ectomycorrhizae. t. aestivum (incl. forma uncinatum) has been recently rediscovered as a valuable alimentary product in many european countries and is currently considered to be the most common european truffle with gradually increasing commercial value. knowledge of the interactions of this truffle species with accompanying soil microflora might be of practical importance, for example in formulation of complex truffle inocula used in artificial host seedlings inoculations, containing beneficial (e.g., mycorrhization-helper) bacteria. materials and methods bacteria were isolated from the t. aestivum ectomycorrhizae (samples 34, 36 and 39 mentioned in gryndler et al., 2012, collected at the locality dominated by carpinus betulus) using dilution plate technique. mycorrhizae were thrice shaken in 50 ml of sterile water. fresh 100 mg aliquots were immediately homogenized in 5 ml sterile water using mortar and pestle and then suspended in 50 ml water. resulting suspension was then diluted by sterile water 1:10 through 1: 10 000 and 25 μl aliquots were spread on the surface of the solid medium a and incubated for 1-4 weeks at 25ºc. medium a contained malt extract (fluka 70167) 5 g, potato extract (fluka 07915) 5g, yeast extract (oxoid l21) 2 g, caco3 1 g, anhydrous cacl2 73 mg, kh2po4 100 mg, kno3 19.3 mg, ca(no3)2.4h2o 292 mg, mgso4.7h2o 196 mg, na2so4 70 mg, k2so4 38 mg, nh4no3 2.5 mg and agar 14 g in one liter, ph (before autoclaving) 7.0. isolation of bacteria 157 the diluted suspension from the sample 36 was incubated also on other 14 different media sharing the following composition of mineral salts: (nh4)2so4 4 g, k2hpo4 2 g, kh2po4 1 g, mgso4.7h2o, agar 10 g per liter, ph 7.0. the organic components of the different media were: yeast extract (0.25%), or casamino acids (0.03%) with yeast extract (0.03%) and glucose (0.03%), or humic acid (0.1%) with vitamins (thiamin-hcl, riboflavin, nicotinic acid, pyridoxin-hcl, inositol, ca-pantothenate, p-aminobenzoic acid, and biotin, each at concentration of 0.5 mg per l), or yeast extract (0.25%) with cellulose powder (4%), or starch (2%), or glycerol (0.05%) with arginine (0.1%), or oak root powder with yeast extract (0.1 %), or oak root extract (0.02%) with yeast extract (0.1%), or colloidal chitin (0.5%), or colloidal chitin (0.5%) with yeast extract (0.1%), or gelatin (1%), or gelatin (1%) with yeast extract (0.1%), or poly-l-lactate (0.1%) with oxgall (0.01%), or poly-l-lactate (0.1%) with oxgall (0.01%) and yeast extract (0.1%). ten replicate plates were established per medium. growing bacteria were subcultured on medium b with the same composition as the above medium a, except that caco3 was omitted and the concentration of anhydrous cacl2 was increased to 1.27 g per liter. bacterial colonies of different morphological properties were chosen for subcultivation in order to obtain as many as possible different bacterial taxa. bacterial isolates were then inoculated to 10 ml of the liquid medium b (without agar) and bacterial biomass was pelleted by centrifugation. dna was extracted from the pellet using nucleo-spin soil dna kit (macherey-nagel gmbh & co., germany) and a fragment of 16s rdna was amplified in pcr with forward primer eub530f (5´-gtg cca gcm gcn gcg g-3´) and reverse primer eub1100ar (5´-ggg ttn cgn tcg ttg cg-3´). the primers were modified from dowd et al. (2008). cycling conditions were 94 ºc for 5 min, followed by 35 cycles of 94 ºc for 1 min, 62 ºc for 50 s, and 72 ºc for 30 s, and concluded by incubation at followed by 72 ºc for 10 min. pcr products were then sequenced and the identity of isolates was estimated using comparison of their partial 16s rdna sequence with genbank database. selected isolates were tested for their possible effects on the growth of culture of t. aestivum, strain tae5 (maintained in the laboratory of fungal biology, institute of microbiology ascr, prague, czech republic). the truffle mycelium was first pre-cultured on the medium pex (pebeyre s.a., cahors, france) for 6 weeks and the bacterium was then applied as one 4-cm line per dish, whose middle was 5 mm apart from the edge of the mycelial colony. the growth of tuber mycelium in the proximity of the bacterial colony was observed after 7 days of co-cultivation. results in total, 183 bacterial isolates were obtained from ectomycorrhizae of t. aestivum. based on the partial sequence of the 16s rdna, they belong to 6 bacterial orders: actinomycetales (139 isolates), burkholderiales (4 isolates), enterobacteriales (1 isolate), pseudomonadales (2 isolates), rhizobiales (33 isolates) and xanthomonadales (4 isolates). among the actinomycetales, distinct groups of isolates were obtained, corresponding to 9 genera: streptomyces (127 isolates), kocuria (1 isolate), microbacterium (1 isolate), micromonospora (3 isolates), nocardia (1 isolate), nocardiopsis 158 m. gryndler and h. hršelová (1 isolate), nonomuraea (2 isolates), rhodococcus (2 isolates) and rothia (1 isolate). no culture of a member of the pseudonocardineae suborder was isolated. the isolates belonging to the order rhizobiales involved the members of the genera phyllobacterium (23 isolates), rhizobium (6 isolates), bosea (1 isolate), ensifer (1 isolate), mesorhizobium (1 isolate) and microvirga (1 isolate). the order pseudomonadales was represented by the genera pseudomonas (possibly p. putida, the only organism willing to grow on the medium containing oak root extract) and moraxella. two members of the order burkholderiales were further isolated: acidovorax sp. (3 isolates) and xylophilus sp. (1 isolate). escherichia (possibly e.coli, 1 isolate) was the only representative of the order enterobacteriales and the order xanthomonadales was represented by the genera lysobacter (possibly l. antibioticus, 3 isolates) and dyella (1 isolate). the identification of the isolates is provisional and may be refined in future, if a particular isolate will prove to have beneficial effects on mycorrhizal colonization of truffle-inoculated tree seedlings. the interactions with culture of t. aestivum were tested for moraxella sp., lysobacter sp., phyllobacterium sp., rhizobium cf. giardinii, rhizobium cf. leguminosarum, mesorhizobium sp., xylophilus sp.,3 isolates of acidovorax sp., microvirga sp. and nocardia sp. some bacterial isolates (rhizobium cf. leguminosarum, rhizobium cf. giardinii, phyllobacterium sp., microvirga sp.) grew vigorously on the pex medium, whereas growth of others (moraxella sp., lysobacter sp., nocardia sp.) was slower. after 7 days of co-cultivation, the only interaction of truffle mycelium with a bacterial isolate was cell vacuolization and dying in proximity of rhizobium cf. leguminosarum (fig. 1). no other case of adverse or stimulatory effects was noted. fig. 1. interaction of in vitro-cultivated mycelium of t. aestivum with phyllobacterium sp. (a, c) or rhizobium cf. leguminosarum (b, d). during the 7-day co-cultivation, the bacteria (dark mass in the figure) became to close contact with hyphae (a, b). whereas hyphae in the proximity of phylobacterium sp. colony had normal morphology (c), the proximity of rhizobium cf. leguminosarum caused extensive vacuolization. scale bar = 100 μm. isolation of bacteria 159 discussion in spite of extensive effort during the isolation of the bacteria from ectomycorrhizae of t. aestivum, we were able to cultivate the members of 6 bacterial orders. this is very low number in comparison with the results of molecular analysis of the same material (gryndler et al. 2012), which detected operational taxonomic units that can be grouped into a total of 79 bacterial orders. this discrepancy confirms the hypothesis that cultivation-based studies reveal just negligible portion of the microbial diversity of mycorrhizosphere. our recently reported effort is the continuation of the past works (gryndler et al. 2012) led by the intention to cultivate mainly the bacterial taxa that are specifically associated with ectomycorrhizae of t. aestivum. this effort was, however, only partially successful and the isolates of lysobacter sp. and ensifer sp. remain the only bacteria that were significantly positively associated with t. aestivum ectomycorrhizae. in particular, we repeatedly failed to isolate the members of the actinobacterial genera actinosynnema, allokutzneria, kibdelosporangium and lentzea, the members of the suborder pseudonocardineae which proved to positively correlate with the presence of t. aestivum in ectomycorrrhizae. in spite of the fact that pseudonocardineae members are generally considered rare organisms in the nature (jarerat et al. 2002), our molecular data predict that pseudonocardineae members represent significant portion of biomass of the root-associated microbial community. however, they either do not produce sufficient amounts of colony forming units necessary for isolation or cannot be cultivated using our methodology. at the same time, their colony forming units may be hidden on dilution plates by high numbers of massively sporulating members of actinomycetales. there is no information on interactions of the members of this suborder with fungi or plants available in the literature. some members of this group possess the ability to degrade poly-l-lactate (a kind of plastic, also used as a cultivation medium component in our work), which was originally considered as unique among the actinobacteria (jarerat et al. 2002). even though some other actinobacterial degraders of this material were described later (sukkum et al. 2009), the suborder pseudonocardineae remains important pool of organisms possessing this activity. this may indicate that the members of this suborder may possess exotic metabolic capabilities, perhaps connected with very specific nutritional demands which were not met during our isolation approach. further work and probably 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(1988) mycorrhizal interactions with the rhizosphere microflora: the mycorrhizosphere effect. phytopathology 78: 366–371. mello a., miozzi l., vizzini a., napoli c., kowalchuk g., bonfante p. 2010. bacterial and fungal communities associated with tuber magnatum-productive niches. plant biosyst. 144: 323–332. murat c., vizzini a., bonfante p., mello a. 2005. morphological and molecular typing of the belowground fungal community in a natural tuber magnatum truffle-ground. fems microbiol. lett. 245: 307–313. napoli c., mello a., bonfante p. 2008. dissecting the rhizosphere complexity: the truffle-ground study case. rend. lincei sci. fis. nat. 19: 241–259. sukkhum s., tokuyama s., tamura t., kitpreechavanich v. 2009. a novel poly(l-lactide) degrading actinomycetes isolated from thai forest soils, phylogenic relationship and enzyme characterization. j. gen. appl. microbiol. 55: 459–467. sbrana c., agnolucci m., bedini s., lepera a., toffanin a., giovannetti m., nuti m.p. 2002. diversity of culturable bacterial populations associated to tuber borchii ectomycorrhizas and their activity on t. borchii mycelial growth. fems microbiology letters 211: 195–202. zacchi l., vaughan-martini a., angelini p. 2003. yeast distribution in a truffle-field ecosystem. ann. microbiol. 53: 275–282. 2014-01-02t12:11:35+0100 polish botanical society agonimia species and other rare lichens in central poland anna łubek institute of biology, jan kochanowski university, świętokrzyska 15a pl-25-406 kielce, anna.lubek@ujk.edu.pl łubek a.: agonimia species and other rare lichens in central poland. acta mycol. 47 (2): 203–212, 2012. eight lichen species are presented. four of them: agonimia flabelliformis, a. opuntiella, a. tristicula and micarea adnata are new to central poland. caloplaca cerina var. chloroleuca, micarea hedlundii and porina leptalea are very rare in the świętokrzyskie mts and in central poland. key words: ascomycota, lichen distribution, protected areas introduction although the świętokrzyskie mts have relatively well known lichen biota, recent explorations provide new species. as a result of these investigations eight species rarely reported from poland have been found, including agonimia flabelliformis, a. opuntiella, a. tristicula, bacidia sulphurella, caloplaca cerina var. chloroleuca, micarea adnata, m. hedlundii and porina leptalea. so far four of them were not recorded in central poland. the species of the genus agonimia deserve special attention. so far, seven species of this genus have been identified in poland (tab. 1): agonimia allobata, a. gelatinosa, a. opuntiella, a. repleta, a. tristicula, a. vouauxii and recently described a. flabelliformis. most of them occur mainly in mountain areas and in northern part of poland. agonimia allobata, corticolous species or found also on plant debris, has been reported at several sites in śnieżnik massif and bialskie mts (szczepańska 2008), pieniny mts (kiszka 2000), gorce mts (czarnota et al. 2005), borecka forest (cieśliński 2003) and mazovia region (kubiak 2009). agonimia gelatinosa is terricolous, on calcareous soil, muscicolous and grows also on plant debris. this rare species in poland is known from kujawy area (ceynowa-giełdon 2001), gorce mts (czarnota 2000) and sudety mts (kossowska 2008). agonimia opuntiella in poland is acta mycologica vol. 47 (2): 203–212 2012 204 a. łubek known mostly from mountain regions, the eastern and western carpathians (kiszka 1997; bielczyk 2003; fałtynowicz 2003) where it grows on calcareous soil and on bryophytes in sunny places. agonimia repleta is more frequent species in this country. it is saxicolous, muscicolous and corticolous species found usually in wet forests on deciduous trees: acer, alnus, fagus and ulmus. it is reported for example from the north-eastern part of the country (czyżewska et al. 2001; cieśliński 2003), from its central part (łubek 2012a, b) and from its south part (czarnota, coppins 2000; kiszka, kościelniak 2001; bielczyk et al. 2004). agonimia tristicula grows on calcareous soil, on bryophytes and on plant debris in open habitats beeing quite common in mountain areas, the western and eastern tatry mts and pieniny mts. only locality in northern part of poland, in the mazurian lake district is also known (olech, kiszka 1999). agonimia vouauxii in this country is reported from anthropogenic habitats from olkusz region (bielczyk 2012). the main aim of this work is to provide the most up-to-date information for the new and rare species in central poland, especially for those recently described. fig. 1. localization of investigated sites: 1 – ‘oleszno’ nature reserve, 2 – ‘świnia góra’ nature reserve, 3 – ‘góra miedzianka’ nature reserve, 4 – ‘zapusty’ slope. agonimia species in central poland 205 material and methods lichens were collected in 2001, 2007, 2010 and 2011 at four sites in the świętokrzyskie mts and in their immediate vicinity (fig. 1): in ‘góra miedzianka’ nature reserve in the chęcińsko-kielecki landscape park, in ‘zapusty’ slope in the świętokrzyski national park, in ‘świnia góra’ nature reserve in the suchedniowsko-oblęgorski landscape park and in ‘oleszno’ nature reserve in the przedborski landscape park, located in squares ee 54, 60, 72, 77 of atpol grid square system (cieśliński, fałtynowicz 1993). the material was analyzed with standard morphological and anatomical methods. results and discussion the nomenclature of the species follows smith et al. (2009) with exception of agonimia flabelliformis – guzow-krzemińska et al. (2012). characteristics of agonimia representatives (tab. 1) according to smith et al. (2009), guzow-krzemińska et al. (2012) and sérusiaux et al. (1999). the specimens are deposited in the herbarium of the jan kochanowski university (ktc). abbreviations: śnp – świętokrzyski national park; s-olp – suchedniowsko-oblęgorski landscape park; przlp – przedborski landscape park, ch-klp – chęcińsko-kielecki landscape park. agonimia flabelliformis halda, czarnota & guzow-krzemińska this species was described from the czech republic, and mentioned additionally from germany and great britain (guzow-krzemińska et al. 2012). in poland it is only known form tatry mts (czarnota 2012). its site in ‘oleszno’ nature reserve is the first in central poland. species is characterized by a small flabelliform thallus and globose perithecia with 8-spored asci. for the full description of the species see guzow-krzemińska et al. (2012). a. flabelliformis resembles a. allobata, from which it differs only in the presence of flabelliform aggregations of goniocysts (guzow-krzemińska et al. 2012). agonimia flabelliformis prefers rather humid, shaded, mossy places within deciduous forests where usually grows on bark and on soil. the collection from the przedborski landscape park was found on the trunk of fraxinus excelsior with the company of bacidia pycnidiata and bacidina sulphurella. it has a dull-green minutely flabelliform squamulose to coralloid thallus, a few grey-brown perithecia which are superficial or partially immersed between squamules. specimen examined. atpol grid square ee 60 − przlp, near the ‘oleszno’ nature reserve, forest section no 79, 50°56’42”n/ 20°06’17”e, on a trunk of fraxinus excelsior, 7 feb. 2008. agonimia opuntiella (buschardt & poelt) vĕzda syn. phaeophyscia opuntiella (buschardt & poelt) hafellner the species occur in many countries in europe such as: belgium, luxembourg, france (diederich et al. 2011), great britain and ireland (smith et al. 2009), the 206 a. łubek czech republic (vĕzda, liška 1999), switzerland (clerc, truong 2010) and germany (wirth et al. 2010). agonimia opuntiella deserves special attention because of the perithecia which are formed very rarely. this species is characterized by greenish-grey to brownish minutely squamulous thallus (lobes up to 4 mm width) with marginal ellipsoid to subglobose blastidia. surface of thallus has minute hyaline hairs. perithecia are formed rarely or if so they are small pyriform, black and rugose (0.2-0.5×0.2-0.4 mm) on the lobe bases (aptroot et al. 2008; aptroot 2011). two densely muriform ascospores, 40-70×20-30 μm, are produced in asci. so far, in poland ascocarps of agonimia opuntiella have not been reported (kiszka 1997). the collection from the chęcińsko-kielecki landscape park has no perithecia indeed, but the specimen from świętokrzyski national park produces a few small, inconspicuous black perithecia, located on the lobe base. they are sometimes partially hidden by adjacent squamules and blend with dark ground decaying bryophytes: more visible when wet. spores of agonimia opuntiella, similarly to a. tristicula, are produced per two in ascus, but they are much smaller. ascospores of a. opuntiella from the świętokrzyski national park have dimensions of 60-63×26-28 μm and in a. tristicula, which was found in the chęcińsko-kielecki landscape park, they reach 72-96×36-48 μm. agonimia opuntiella from the świętokrzyski national park was already published, however under the name phaeophyscia kairamoi – young thallus in the initial stage of growth was difficult to determine (łubek 2003; łubek, cieśliński 2004; łubek 2007). after a careful examination of the specimen, a few small perithecia have been found, thus now there is no doubt that it represents a. opuntiella. these two species are similar because of the presence of marginal blastidia and tiny hyaline hairs on the surface of thallus. because of this mistake, ph. kairamoi should be deleted from the list of lichens of the świętokrzyskie mts. here a. opuntiella is reported also as new to central poland. specimens examined. atpol grid square ee 72 − ch-klp, ‘góra miedzianka’ nature reserve, on bryophytes growing on calcareous rocks, s exposure, 50o50’44”n/ 20o2’46”e, 8 oct. 2011, leg. b. słowińska (ktc); ee 77 – śnp, ‘zapusty’ slope, on bryophytes growing on calcareous rocks, s exposure, 50o53’51”n/ 21o06’09”e, 31 aug. 2001. agonimia tristicula (nyl.) zahlbr syn. verrucaria tristicula nyl. this species occurs in many countries in europe such as: belgium, luxembourg, france (diederich et al. 2011), the czech republic (vĕzda, liška 1999), estonia (randlane, saag 1999), sweden and norway (santesson 1993), switzerland (clerc, truong 2010) and germany (wirth et al. 2010). agonimia tristicula is reported here as new to the świętokrzyskie mts and central poland. it was found in the chęcińsko-kielecki landscape park, where grew on bryophytes on calcareous soil. it characterizes by dull to pale green or brown minutely squamulose thallus, which often forms small, aggregate patches. perithecia are frequent, fairly big, barrel-shaped, black, matt and plicate-rugose. asci produce two spores: 72-96×36-48 μm. specimen examined. atpol grid square ee 72 − ch-klp, ‘góra miedzianka’ nature reserve, on bryophytes growing on calcareous rocks, s exposure, 50o50’44”n/ 20o21’46”e, 8 oct. 2011, leg. b. słowińska. agonimia species in central poland 207 bacidina sulphurella (samp.) m. hauck & v. wirth syn. bacidia sulphurella samp. in europe this species is known from many countries: belgium, luxembourg, france (diederich et al. 2011), great britain and ireland (smith et al. 2009), switzerland (clerc, truong 2010) and germany (wirth et al. 2010). in poland bacidina sulphurella is known for example from: góry sowie mts and puszcza knyszyńska forest (brand et al. 2009), gorce mts (czarnota 2010) and pogórze wiśnickie foothills (śliwa 2010). in central poland the species was reported from few localities in ‘las bielański’ forest reserve (kubiak et al. 2010) and wzniesienia łódzkie landscape park (hachułka 2011). it is surely more common species but formerly many times determined as bacidina arnoldiana (see czarnota 2010). bacidina sulphurella is characterized by finely granular, green thallus and white, pale to greyish coloured, usually present pycnidia, producing filiform, at least in one end strongly hooked, 0-3 septate conidia (brand et al. 2009). the specimen from the ‘oleszno’ nature reserve has few typical pycnidia and well developed thallus. the species grow in the company of agonimia flabelliformis. specimen examined. atpol grid square ee 60 − przlp, near ‘oleszno’ nature reserve, forest section no 79, on a trunk of fraxinus excelsior, 50°56’42”n/ 20°06’17”e, 7 feb. 2008, (ktc, with agonimia flabelliformis and bacidia pycnidiata), det. p. czarnota. caloplaca cerina var. chloroleuca (sm.) th. fr. syn. caloplaca cerina var. muscorum a. massal. this is a very rare species in poland (fałtynowicz 2003). it is known primarily from mountain areas: sudety mts, western carpathians mts and wyżyna śląskokrakowska upland (fałtynowicz 2003) as well as from the north-eastern part of poland (cieśliński 2003) where grows on plant debris, soil or on mosses in xerothermic grasslands and gravel pits. in central poland caloplaca cerina var. chloroleuca is known from zelejowa mt. near ‘skorocie’ nature reserve and near lasocin village in the świętokrzyskie mts (cieśliński 1979). specimen of c. cerina var. chloroleuca collected in the chęcińsko-kielecki landscape park is small and has a scurfy, green-grey thallus, and a few sessile apothecia with thick, grey thalline margin and orange-yellow disc. specimen examined. atpol grid square ee 72 − ch-klp, ‘góra miedzianka’ nature reserve, on bryophytes growing on calcareous rocks, s exposure, 50o50’44”n/ 20o21’46” e, 8 oct. 2011, leg. b. słowińska. micarea adnata coppins this is the first information on the occurrence of this species in the świętokrzyskie mts and in central poland. in this country it is known only from two mountain regions: karkonosze mts (kossowska 2001) and western beskidy mts (czarnota 2007). in europe this species is widespread; known from the czech republic (vĕzda, liška 1999), sweden (santesson 1993), switzerland (clerc, truong 2010), germany (wirth et al. 2010) and other countries (see czarnota 2007). micarea adnata is easily to identify owing to its characteristic apothecia and sporodochia (czarnota 2007). it grows on the bark of coniferous trees, on wood or decaying stumps. the specimen mentioned here has numerous straw-coloured apothecia, which are immarginate or sometimes surrounded with distinctly visible white 208 a. łubek ta bl e 1 c om pa ri so n of s el ec te d fe at ur es o f p ol is h re pr es en ta ti ve s of th e ge nu s a go ni m ia c ha ra ct er is ti cs o f t he s pe ci es a cc or di ng to : 1 sm it h et a l. (2 00 9) ; 2 g uz ow -k rz em iń sk a et a l. (2 01 2) ; 3 sé ru si au x et a l. (1 99 9) fe at ur e a go ni m ia a llo ba ta 1 a go ni m ia fla be lli fo rm is 2 a go ni m ia ge la tin os a1 a go ni m ia op un tie lla 1 a go ni m ia re pl et a1 a go ni m ia tr is tic ul a1 a go ni m ia v ou au xi i3 t ha llu s gr ey -g re en to br ow n, c on ti nu ou s an d m in ut el y ro ug he ne d to g ra nu la r, so m et im es co m po se d of b ra nc he d st ru ct ur es du llgr ee n m in ut el y fla be lli fo rm to sq ua m ul os e da rk b ro w n co m po se d of r ou nd ed or lo be d go ni oc ys ts , pr ot ha lu s co ns pi cu ou s gr ee ni sh -g re y to br ow ni sh , m in ut el y of s qu am ul es to 4 m m d ia m ., m ar gi na l e lli ps oi d to s ub gl ob os e bl as ti di a, m in ut e hy al in e ha ir s on su rf ac e of th al lu s du ll gr ee n, gr an ul ar , gr an ul ar ve rr uc os e or m in ut el y sq ua m ul os e du ll to p al e gr ee n or b ro w n, m in ut el y sq ua m ul os e, fo rm in g sm al l, sp re ad in g, ag gr eg at e pa tc he s gr ee ni sh to b ro w ni sh , gr an ul os e to sq ua m ul os e, p ap ill at e, w it h sq ua m ul es o f 0 ,6 -2 m m d ia m . pe ri th eci a fr eq ue nt , g lo bo se or ta lle r th an w id e; s ur fa ce : gr ey -b ro w n to bl ac k, s m oo th , m at t fr eq ue nt , g lo bo se , su pe rfi ci al or p ar ti al ly im m er se d be tw ee n sq ua m ul es ; su rf ac e: g re ybr ow n, s m oo th an d m at t fr eq ue nt , gl ob os e, su bs ph er ic al ; su rf ac e: sm oo th , b la ck , m at t ve ry r ar e, p yr if or m , on th e lo be b as es ; su rf ac e: b la ck , ru go se fr eq ue nt , py ri fo rm , h al f t o th re equ ar te rs im m er se d; su rf ac e: g re ybr ow n to b la ck , ro ug he ne d w it h ve rt ic al c ra ck s ra th er fr eq ue nt , fa ir ly b ig , b ar re lsh ap ed , b et w ee n an d ov er gr ow n by s qu am ul es ; su rf ac e: b la ck , m at t a nd p lic at eru go se fr eq ue nt , s ub sp he ri ca l to s lig ht ly o bp yr if or m , ha lf -i m m er se d; s ur fa ce : bl ac ki sh , g lo ss y si ze o f pe ri th eci a 12 -2 2 μm d ia m . 15 -2 5 μm d ia m . 30 -5 0 μm di am . 20 -5 0 μm d ia m . 14 -2 0 μm d ia m . 24 -5 0 μm d ia m . 12 -2 3 μm d ia m . sp or es 8 in a sc us , 2 935 × 10 -1 5 μm , m ur if or m 8 in a sc us , 2 335 × 11 -1 5 μm , m ur if or m 8 in a sc us , 3 155 × 15 -2 0 μm , m ur if or m 2 in a sc us , 4 070 × 20 -3 0 μm , de se nt ly m ur if or m 8 in a sc us , 2 046 × 12 -2 0 μm , m ur if or m (1 -) 2 in a sc us , 5 712 0× 26 -5 0 μm , m ur if or m 2 in a sc us , 6 072 × 15 -2 4 μm , m ur if or m su bs tr ate o f oc cu rre nc e in e ur op e ba rk ( q ue rc us , u lm us , a ln us , fr ax in us , a ce r, p in us , p ic ea , t ili a, c ar pi nu s) , p la nt de br is ba rk ( p ic ea , fr ax in us , f ag us , t ili a, l ar ix , q ue rc us ), de ca yi ng w oo d, ro ck s ca lc ar eo us s oi l, br yo ph yt es , pl an t d eb ri s ca lc ar eo us s oi l, br yo ph yt es , r ar el y on b ar k (q ue rc us ) ba rk ( a ce r, a ln us , f ag us , u lm us , p ic ea ), br yo ph yt es o n ro ck s ca lc ar eo us s oi l, in c re vi ce s of ca lc ar eo us r oc ks , br yo ph yt es , p la nt de br is , r ar el y on ba rk ( a ce r, u lm us , fr ax in us , a ln us , so rb us ) br yo ph yt es , p la nt de br is , s oi l h ab it at re qu ir em en ts m oi st p la ce s in de ci du ou s fo re st hu m id , s ha de d an d m os sy p la ce s in d ec id uo us to m ix ed fo re st sh ad y an d m oi st p la ce s in c al ca re ou s ha bi ta ts su nn y pl ac es in ro ck s gr as sl an ds m oi st p la ce s in d ec id uo us fo re st s su nn y pl ac es in xe ro th er m ic a nd ro ck s gr as sl an ds th er m op hi lo us gr as sl an d, o pe n ha bi ta ts of li m es to ne a re as a nd an th ro po ge ni c ha bi ta ts agonimia species in central poland 209 rim. convex or globose sporodochia, resembling small apothecia, are numerous and produces macroconidia 8-9×2-3 μm. due to the specific habitat requirements and its occurrence only in natural forests, m. adnata is proposed to be regarded as an indicator of ecological continuity for woodlands of lower mountain belt in central europe (czarnota 2007). also the site in central poland is located in such natural forest. ‘świnia góra’ nature reserve is one of the oldest in the świętokrzyskie mts, being remnants of the puszcza świętokrzyska forest. specimen examined. atpol grid square ee 54 − s-olp, ‘świnia góra’ nature reserve, on lignum, 51o03’16”n/ 20o41’58”e, 10 nov. 2010. micarea hedlundii coppins in poland it is reported mainly from eastern and southern poland (czarnota 2007). in central poland micarea hedlundii was already recorded in the świętokrzyski national park (łubek 2003), in the ‘spała’ nature reserve and the puszcza kozienicka forest (czyżewska et al. 2005; czarnota 2007). characteristic feature of this species are pycnidia coated with white tomentum and orange droplets of an oil substance in goniocysts which reacting k+ purpleviolet. specimen examined from the suchedniowsko-oblęgorski landscape park has only pycnidia. specimen examined. atpol grid square ee 54 − s-olp, ‘świnia góra’ nature reserve, on lignum, 51o03’16”n/ 20o41’58”e, 10 nov. 2010. porina leptalea (durieu & mont.) a. l. sm. this species is known from many european countries, for example: belgium, luxembourg, france (diederich et al. 2011), switzerland (clerc, truong 2010), germany (wirth et al. 2010) and the czech republic (vĕzda, liška 1999). in poland is a rare species found in mountain areas: gorce mts (czarnota 2000), pieniny mts (kiszka 2000), bieszczady niskie mts (kościelniak 2004) and sudety mts (szczepańska 2007). in the świętokrzyskie mts it seems to be also rare. its first four sites in the świętokrzyski national park have already been published (łubek 2003) and here is presented additional locality of this species in this region. specimen examined has dull orange-brown to red-brown perithecia, hemispherical to subglobose 0,1-0,4 mm diam., which are one quarter to one half immersed in thin grey-green thallus. ascospores are fusiform-cylindrical or oblong, 3-septate and 15-25×2-4 μm. it has also orange pycnidia with narrowly ellipsoid conidia 3-4,5×1 μm. specimen examined. atpol grid square ee 54 − s-olp, ‘świnia góra’ nature reserve, on trunk of fagus sylvatica, 51o03’16”n/ 20o41’58”e, 10 nov. 2010. conclusions the świętokrzyskie mts appear to be very interesting from lichenological point of view. the specific topography, various geology and often natural vegetation, make appropriate conditions for lichens both widespread and of narrow ecological-scale. particularly noteworthy are the species occurring in poland only in mountain areas, including agonimia opuntiella, micarea adnata and porina leptalea. 210 a. łubek among the studied areas noteworthy are ‘oleszno’ nature reserve in the przedborski-landscape park and ‘świnia góra’ nature reserve in the suchedniowskooblęgorski landscape park. the natural character of forests in the ‘oleszno’ nature reserve has already been underlined few times by the presence of very rare epiphytic lichens in poland (łubek 2009a, b). in this paper two next lichen-forming fungi are interesting: agonimia flabelliformis and bacidina sulphurella. first one because up to now it was found in poland only once, and the second species because its real distribution in europe is still poorly known. ‘świnia góra’ nature reserve appears to be also noteworthy protected area owing to a large amount of dead wood inhabited by lignicolous lichens, such as: micarea adnata and m. hedlundii. acknowledgements. i would like to thank professor krystyna czyżewska (university of łódź) for many valuable suggestions received during the preparation of the manuscript, professor paweł czarnota (scientific laboratory, the gorce national park) for determining bacidia sulphurella and confirmed the determination of agonimia flabelliformis and micarea adnata, and beata słowińska for common filed works. sincere thanks are also addressed to anonymous reviewers for considerable remarks on the text. references aptroot a. 2011. new lichen records from australia 73. agonimia opuntiella. australasian lichenology 68: 3. http://www.nhm.uio.no/botanisk/lav/rll/al/al68.pdf aptroot a., lücking r., sipman h.j.m., umańa l., chaves j.l. 2008. pyrenocarpous lichens with bitunicate asci. a first assessment of the lichen biodiversity inventory in costa rica. bibliotheca lichenologica 97: 1–162. http://www.inbio.ac.cr/papers/liquenes/pdf/pyrenocarpos_2008.pdf bielczyk u. 2003. the lichens and allied fungi of the polish western carpathians. 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(in:) s. cieśliński, w. fałtynowicz (eds). atlas of the geographical distribution of lichens in poland 2: 7–11. w. szafer institute of botany, polish academy of science, kraków. sérusiaux e., diederich p., brand a.m., van den boom p. 1999. new or interesting lichens and lichenicolous fungi from belgium and luxembourg. viii. lejeunia 162: 1–95. szczepańska k. 2008. antropogeniczne przemiany bioty porostów masywu śnieżnika i gór bialskich. acta bot. siles., monogr. 4: 1–291. 212 a. łubek smith c.w., aptroot a., coppins b.j., fletcher a., gilbert o.l., james p.w., wolseley p.a. 2009. the lichens of great britain and ireland. the british lichen society, london, 1046 pp. śliwa l. 2010. contribution to the lichen biota of the pogórze wiśnickie foothills (carpathians). acta mycol. 45 (2): 219–230. vězda a., liška j. 1999. katalog lišejníků české republiky. institute of botany, academy of sciences of the czech republic, průhonice, 283 pp. wirth v., hauck m., von brackel w., cezanne r., de bruyn u., dürhammer o., eichler m., gnüchtel a., litterski b., otte v., schiefelbein u., scholz p., schultz m., stordeur r., feuerer t., heinrich d., john v. 2010. checklist of lichens and lichenicolous fungi in germany. georg august university of göttingen, germany. http://www.gwdg.de/~mhauck gatunki z rodzaju agonimia i inne rzadkie porosty w polsce środkowej streszczenie w pracy przedstawiono osiem gatunków porostów: agonimia flabelliformis, a. opuntiella, a. tristicula, bacidina sulphurella, caloplaca cerina var. chloroleuca, micarea adnata, m. hedlundii i porina leptalea, które stwierdzono ostatnio w górach świętokrzyskich i ich najbliższym sąsiedztwie. wśród nich, cztery nie były dotąd znane z tego pasma górskiego i z polski środkowej. są to: agonimia flabelliformis, a. opuntiella, a. tristicula i micarea adnata. badane materiały zdeponowane są w zielniku uniwersytetu jana kochanowskiego w kielcach. 2014-01-02t12:15:59+0100 polish botanical society mycological monitoring in the hungarian biodiversity monitoring system ferenc pál-fám1, irén siller2 and lívia fodor3 1university of kaposvár, department of botany and plant production h-7400 kaposvár, guba s. 40., pff3@hotmail.com 2szent istván university, faculty of veterinary sciences, department of botany h-1077 budapest, rottenbiller 50., turcsanyine.siller.iren@aotk.szie.hu 3state secretariat of nature and environment protection, ministry of environment and water h-1011 budapest, fő u. 44-50., kisne@mail.kvvm.hu p á l f á m f., s i l l e r i., f o d o r l.: mycological monitoring in the hungarian biodiversity monitoring system. acta mycol. 42 (1):35-58, 2007. the results of a mycological monitoring, carried out from 2001 until 2003 in two forest reserves (in the bükk and the mecsek mountains) within the frame of a project of the hungarian biodiversity monitoring system (hbms) aiming to monitor forest reserves and managed forests, are presented. standard sampling method had to be developed and methods of data analysis (a so-called protocol) had to be elaborated for monitoring activities. key words: macromycetes, fungal monitoring, forest reserves, managed forests, plantations introduction fungi play a very important role in the circulation of matter in ecosystems. therefore the group of fungi has been also chosen as a component of hbms. the number of monitoring activities including fungi is relatively low, and the methodology of sampling is not elaborated properly. for this reason thorough investigations and analyses were needed during the development of the monitoring program. monitoring means a series of long-term, systematic studies carried out by standardized methods. during elaboration of protocols including the detailed methodological description of sampling and data processing numerous questions have arisen, which demanded new approaches from mycological experts besides application of classical methods. the work on the methodological basis of fungal monitoring, which, in the meantime, was modified several times according to the collected experiences (p á l f á m 1999; r i m ó c z i et al. 2000), was started in 1999 within the framework of hbms (r i m ó c z i , p á l f á m 1999). acta mycologica vol. 42 (1): 35-58 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 36 f. pál-fám et al. due to the special life cycle of fungi, choosing among the monitoring methods is a difficult task. qualitative and quantitative study methods of macrofungi differ from those used in botany or zoology. mycological monitoring is primarily based on observation of fruit bodies, because vegetative mycelium and mycorrhiza are living in the soil and they are difficult to access and to identify. the study of fruit bodies, however, is not without problems either: their life span is short, their formation is periodical, showing fluctuation and strong dependence on weather, and their ecological functions are very much variable. monitoring of fungal communities is furthermore hampered by numerous taxonomical problems and the lack of experts. the question of data processing meant a big challenge, too. within the frame of hbms the protocols include methods of establishment of so-called derived data for every group of relevant living beings. however, because of the novelty of fungal monitoring, it was supposed that in the course of evaluation of data, besides the simple variables and indexes, application of diversity measurements and multivariate methods can also help in answering questions arisen and in tracing of changes and trends of fungal communities. in the case of fungi many new proposals were put forward in this field, applicability of which can be decided only after a longer time and on the basis of repeated studies. monitoring of fungal communities is realized within the framework of hbms in the areas of forest reserves. in addition to monitoring of conditions of selected habitats of seminatural state (core areas of forest reserves), tracing of effects of forest management was our intention, too. according to the basic hypothesis of the project, forest management modifies species richness of deciduous forests. in order to settle this question monitoring was carried out in the core areas of the selected forest reserves, in their buffer zones (or managed forests) and also in cultivated forests planted nearby the selected forest community. mycological monitoring – after preliminary studies of several years – started in 2001 in two forest reserves in the bükk and the mecsek mountains. on the basis of methods elaborated during the course of preliminary studies the presence of fungi was monitored in permanent sampling plots during periods of 3 years. this length of monitoring period is necessary, because weather dependent appearance of fruit bodies can cause significant differences in the presence of species in consecutive years. that is the reason why summarized results of several years’ examination can be considered as one sample. the sizes of sampling plots were determined on the basis of available results and experiences of mycocoenological studies (p á l f á m 1999, 2001a,b). goals the present work aims to demonstrate the first results of biodiversity monitoring of fungal communities in hungary, and tries to answer some basic questions influencing the further process of monitoring. the goals of monitoring are: 1. identification and assessment of the conservation value of the fungal species in the examined areas; 2. long-term follow-up of characteristics and changes of fungal communities in the involved forests; mycological monitoring 37 3. comparative monitoring of fungal communities in non-managed core areas of forest reserves, managed stands and forest plantations, in order to examine the effects of management in the latter ones; 4. comparative study of fungal communities of forests situated in different geographical regions and on different base rocks; 5. analysis and comparison of monitored areas in terms of different variables (species number, species diversity, fruit body number, functional diversity) indicating characteristics and changes of fungal communities. a reliable clarification of several questions is not yet possible. for trend monitoring of fungal communities of forests long-term monitoring is needed, by the means of which a comparison of summarized results of different samples covering several years’ periods will become possible. for commencing mycological monitoring two forest reserves were chosen in which mycological research had been in progress earlier. the work was performed in the “őserdő” forest reserve in the bükk mountains and in the “kőszegi-forrás” forest reserve of the mecsek mountains, by the same sampling protocol. mycological research of forest reserves in the 1980’s several mycologists participated in regular surveys of german forest reserves (k r i e g l s t e i n e r 1982; m a t h e i s 1985; h a a s , k o s t 1985; w i n t e r h o f f 1989). ve e r k a m p and k u y p e r (1993) completed a mycological survey of sixteen dutch forest reserves, proving the outstanding role of lignicolous fungal species in these habitats. the north european mycological school departed from the western and central european ones. its attention was focused on species inhabiting dead wood (r e n v a l l et al. 1991; r e n v a l l 1995; h ø i l a n d , b e n d i k s e n 1996; s t o k l a n d 2001). they found, that biodiversity of lignicolous fungi (aphyllophorales) living in boreal pine forests was influenced by the degree of continuousness of dead wood supply, the distribution pattern of available dead wood, the effects of forest management and the degree of fragmentation of landscape. forest management and the age of the forests influence species supply, functional spectrum and diversity of fungal communities. these characteristics can be connected mainly with the quantity and quality of dead wood (trunks, logs, etc.) (k o s t , h a a s 1989; s i l l e r 2004). this same was formulated by a r n o l d s as well (1988); according to him aging of forests definitely contributes to the raising number of lignicolous fungi, because the quantity of substrates (branches, logs) available rises. in hungary, the results of mycological surveys in the kékes forest reserve were published by s i l l e r (1986, 1999) and s i l l e r et al. (2002). the fungal communities of the kőszegi-forrás forest reserve of the mecsek mountains and of the őserdő forest reserve in the bükk mountains were compared by s i l l e r et al. (2004). the wood decomposing fungi of the ropolyi forest reserve were surveyed by tr e c z k e r and s z a b ó (2002). s i l l e r et al. (2004) studied the indicator values of fungal communities in comparison with developmental processes of the forests. s i l l e r (2004) revealed different biodiversity indicator species in forest reserves. f o d o r and p á l f á m (2000) analyzed functional spectra of fungal communities in different geographical regions (mecsek, szigetköz). 38 f. pál-fám et al. s i l l e r and m a g l ó c z k y (2002) summarized the methods of mycological surveys in a methodological series of forest reserve research in hungary. the authors proposed different research levels and methods (minimum program, long-term study series, aim oriented research) for the mycological study of forest reserves. among aspects of data evaluation, in addition to the traditional mycocoenological methods they draw attention to the ordination and classification techniques, which are still less used in mycology today. taking into consideration the bioindicative properties of fungi and the occurrence of red list species offers further possibilities. within the framework of comprehensive forest reserve research an opportunity arises to compare results of mycological studies with data of forest structure, soil, microclimate, phytocoenological and forestry investigations. monitoring in mycology there are only a few examples of long-term, regular mycological monitoring activities based on standardized methodologies. in several cases under the expression of mycological monitoring some repeated surveys or the comparison of the results of two sampling periods are meant. under the guidance of a r n o l d s (1988) and a r n o l d s and ve e r k a m p (1999), following some previous comparative analyses, monitoring of 110 mushroom species were accomplished. the estonian biodiversity monitoring system also aimed at following up habitats of 11 endangered species (s o d e r m a n n 1998), but there is no information available about the implementation of the plans. mycorrhizal fungi were used as indicators of the impacts of air pollution (f e l l n e r 1989; f e l l n e r , s o u k u p 1991). d u d k a et al. (1994) carried out mycological monitoring in ukraine in order to predict the future state of forests. similarly, the results of an eight-year study program performed in italy, aiming at following up the health condition of forests, was reported by a m b r o s i et al. (2002). they measured the biomass of macrofungi in permanent plots. nowadays, several researchers (e.g. l i s i e w s k a , p o ł c z y n s k a 1998; -wo j e w o d a et al. 1999; ł a w r y n o w i c z et al. 2001; ł a w r y n o w i c z 2001) return to the forests studied before with the goal of monitoring. s k i r g i e ł ł o (1998) studied fungal communities of the forests of białowieża national park in permanent plots during 3 years, and then she compared the data with the results of a mycocoenological study, which had been performed 40 years earlier. several countries plan to include fungi in their biodiversity monitoring programs and claim this work to be under elaboration. on the basis of the reviewed literature the conclusion can be drawn that the amount of experiences available on standard sampling methods of mycological monitoring and the possibilities of data processing are rather limited. consequently, the results of the present work may bridge a gap and can be expected to serve as a guidance for researchers of other countries as well. mycological monitoring 39 materials and methods description of sampling areas the selected plant community was a montane beech forest (aconito-fagetum) in the bükk mountains and a sessile oak–hornbeam forest (asperulo taurinae-carpinetum) in the mecsek mountains. parallel sampling was performed in the core areas of these forest reserves, and in addition, in two managed forests and plantations each in the close vicinity of the reserves. sampling areas at the őserdő forest reserve in the bükk mountains. climate of the bükk mountains is cool with a montane character. annual average temperature is 6–7.5 °c, annual precipitation is 800–900 mm. the yearly number of frosty days is above 120, the average temperature in july is 16 °c. soil of the monitored forest reserve is brown forest soil with some podsolization in some places. the soil has a ph of 5.9 on average. the őserdő forest reserve is situated in the bükk mountains. in an area of 59.3 hectares it is one of the oldest montane beech (aconito-fagetum) forests in the country and is protected since 1942. long time ago it was managed, but forestry management has not occurred for more than 100 years. in the forest, containing also about 180–200 years old tree giants, natural forest developmental processes can be observed. canopy of the forest is consisted of beech (fagus sylvatica) and – sporadically and mainly in the marginal parts – maples (acer pseudoplanatus, a. platanoides) and common ash (fraxinus excelsior). in the shrub layer – besides the regrowth of the tree species – rhamnus cathartica, rosa canina, sambucus nigra and ribes uvacrispa can be found. the herb layer is relatively poor, represented only by 61 species (k á r á s z , s u b a 1982). in this layer characteristic plant species of beech forests can be found, as galium odoratum, impatiens noli-tangere, oxalis acetosella, glechoma hirsuta, hordelymus europaeus, sanicula europaea and viola sylvestris. montane species of the herb layer are scopolia carniolica, senecio nemorensis subsp. fuchsii, daphne mesereum and lunaria rediviva. core area (b1). a part of the forest reserve being in collapsing phase (with logs and stumps of several collapsed trees in different phases of decay), about the fungal communities of which several years’ data are available. managed forest (b2). this forest used to be under forestry management (by selective felling), but has been undisturbed for 20–25 years. wood material of the last felling was left piled up and fallen branches can be found on the ground surface. its age is approximately 50–60 years and besides the beech (fagus sylvatica) common ash (fraxinus excelsior) and maples (acer pseudoplanatus, a. platanoides) are present in substantial numbers. spruce forest (piceetum cult.) (b3). the plantation was planted into the place of a beech forest. a few beech trees can be found even today in the margins. the picea abies trees are 40–60 years old. at present it is a completely closed, “nudum” forest. the last thinning – similarly to the managed stand – was carried out 20–25 years ago. shrub layer can be found only in the marginal parts, consisting mainly of sambucus nigra. on the ground surface some trunks, logs and branches of collapsed trees can be also found. 40 f. pál-fám et al. sampling areas at the kőszegi-forrás forest reserve in the mecsek mountains. climate of the mecsek mountains has a submediterranean character. the average annual precipitation is 700 mm. there are two peaks of precipitation: in may–june with 60–80 mm and in october–november with 60–78 mm. a brown forest soil developed on limestone is typical of the mountains. the ph of the soil in the monitored area has a value of 4.6–4.8 in the upper 10 cm, whereas with a value of 5.1 in the plantation it is a little bit higher. all forests monitored were in the potential habitat of the sessile oak–hornbeam community (asperulo taurinae-carpinetum) of the mecsek mountains. these forests have the largest extension in mecsek. because a managed forest of the chosen community can not be found in the vicinity of the reserve, the nearest sessile oak–hornbeam forest was marked out for comparison. their characteristic species are sessile oak (quercus petraea), turkey oak (q. cerris) and hornbeam (carpinus betulus). very few exemplars of silver linden (tilia tomentosa), wild cherry (cerasus avium) and maples (acer campestre, a. platanoides) occur also dispersedly. characteristic shrubs are staphylea pinnata and ruscus aculeatus. in the herb layer melica uniflora, carex pilosa, galium odoratum and aegopodium podagraria form facies. typical species of the community are furthermore helleborus odorus, doronicum orientale and lonicera caprifolium. in the core area (m1) and the managed forest (m2) the canopy consists of sessile oak (quercus petraea), turkey oak (q. cerris) and hornbeam (carpinus betulus). beech (fagus sylvatica), field maple (acer campestre), wild cherry (cerasus avium) and the submediterranean floral element silver linden (tilia tomentosa) can be also found dispersedly. the shrub layer is mainly consisted of the regrowth of the species listed above. in the plantation (m3) the dominant species is scots pine (pinus sylvestris). the shrub layer consists of well developed exemplars of rubus spp. in the herb layer carex sylvatica and melica uniflora form facies. the core area is a 100–160 years old, non-managed forest, in which – especially in the stream valley and around a spring – several collapsed or standing decaying trees and regrowth of several ages can be found. the core area of 33.2 hectares is not managed in any way. the managed and the planted forests are about 30 years old. in these thinning occurs every year and the dead wood is transported away. field sampling field sampling was carried out on the basis of the protocol of hbms. fungi develop fruit bodies in different periods of the year. in order to manage to register a substantial proportion of species – depending on precipitation – 5–8 samplings per year are necessary mainly in the summer (june–august) and autumn (september– october) aspects. since fruit bodies of a given species in a habitat even in the course of several years can be absent, while its mycelium is present in the substrate, data of one year can not be taken as a complete sample. for this reason, in our monitoring work 3 years’ data were considered as one sample. borders of the sampling areas were determined by the extensions of the chosen plant communities. consequently, sampling areas have different sizes (0.1–316 hectars, see below). species lists were prepared at every visit of the investigated areas; mycological monitoring 41 furthermore qualitative assessments in permanent plots of 500 square meters were also carried out. all fruit bodies of every species were counted. besides names of species and fruit body numbers the substrates were recorded, too. in this respect plots represented the complete forests. species names were used following the work of k r i e g l s t e i n e r (1991–93). table 1 contains sampling dates. ta b l e 1 sampling dates at the two sites bükk mts (őserdő) mecsek mts (kőszegi-forrás) 10.04.2001 01.05.2002 01.05.2003 02.06.2001 15.06.2002 12.06.2003 16.06.2001 31.05.2002 31.05.2003 03.07.2001 28.06.2002 30.06.2003 28.07.2001 23.06.2002 28.06.2003 18.09.2001 02.09.2002 28.07.2003 29.08.2001 21.07.2002 05.09.2003 01.10.2001 20.09.2002 16.09.2003 23.09.2001 20.08.2002 21.10.2003 02.11.2001 20.10.2002 04.10.2003 29.10.2001 29.09.2002 19.10.2003 31.10.2002 data analysis main methods of data processing were as follows: 1. monitored areas and sampling plots were compared on the basis of species lists and fruit body numbers of fungal communities. 2. compositions of the fungal communities in the sampling plots, as suggested by the work of a r n o l d s et al. (1995), were compared on the basis of life form spectra (functional spectra) concerning data of the separate years and the total of three years alike. species were classified into the following functional categories: st = soil-inhabiting saprotroph, pn = necrotrophic parasite, m = mycorrhizal, sh = wood-inhabiting saprotroph. the basis of comparison was the number of species belonging to the different groups. 3. data representing the numbers of fruit bodies of soiland wood-inhabiting species were analyzed separately because of the different properties of these groups. a) rank-abundance curves prepared of the data of the different sampling plots were compared as suggested by w h i t t a k e r (1970). b) scale-dependent characterization of diversity, based on the data of sampling plots, was carried out by rényi’s generalized entropy-type diversity profile calculation, which can be used for communities of small and big numbers of species alike (t ó t h m é r é s z 1993). in every case the results of rts (right-tail-sum) diversity profiles (t ó t h m é r é s z 1997) were examined, too. 4. by application of principal coordinate analysis with jaccard’s distance function (syntax computer program, p o d a n i 2001) based on cumulative three-year presence-absence data of macrofungi in the investigated forests, the composition of the fungal communities of the different sampling areas were compared. 5. evaluation of the conservational values of fungi was completed on the basis of the proposed red list of hungarian macrofungi (r i m ó c z i et al. 1999). names of species are mostly according to m o s e r (1983). 42 f. pál-fám et al. results and discussion characterization of fungal communities in the core area in the bükk mountains the following species were recorded in relatively large amounts: bjerkandera adusta, datronia mollis, fomes fomentarius, ganoderma lipsiense, marasmius alliaceus, mycena arcangeliana, m. crocata, panellus stypticus, pleurotus pulmonarius, polyporus varius, schizophyllum commune, stereum hirsutum, xylaria hypoxylon, laxitextum bicolor, a cup fungus (ascocoryne cylichnicum) and the microfungus fuligo septica. among saprotrophic species the following ones reached high abundances: clitocybe candicans, c. fragrans, c. phyllophila, c. cfr. obsoleta, c. phaeophtalma, collybia butyracea and collybia peronata. for mycorrhizal species, amongst which lactarius subdulcis was prominent, generally a lower abundance was typical. frequent xylophagous species of the managed forest are crepidotus applanatus, c. variabilis, mycena arcangeliana and psathyrella conopilus. for the plantation typical were mycena arcangeliana and stereum sanguinolentum (xylophags) as well as collybia confluens and c. maculata (saprotrophes). in the core area from among aphyllophorales such rare species were found as antrodiella hoehnelii, a. semisupina, ceriporia purpurea, ceriporiopsis gilvescens, cerrena unicolor, hericium coralloides, ganoderma pfeifferi, skeletocutis nivea, spongipellis pachyodon and ischnoderma resinosum. from among cap fungi also several rare species appeared in the monitored areas. examples are hydropus subalpinus, entoloma dichroum, flammulaster limulatus, ossicaulis lignatilis and some rare pluteus spp. the managed forest and the plantation are very poor in rare species. in the core area of the kőszegi-forrás forest reserve of the mecsek mountains trichaptum biforme had a high abundance. bjerkandera adusta, panellus stypticus and hypholoma fasciculare were also frequent. none of soil-inhabiting saprotrophs produced a significant amount of fruit bodies and among mycorrhizal ones only lactarius camphoratus is worth mentioning. frequent species of the managed forest are also xylophags. such are stereum hirsutum, stereum subtomentosum, hypoxylon fragiforme, schizophyllum commune, hypholoma fasciculare and panellus stypticus. the plantation – besides hypholoma fasciculare and the soil-inhabiting saprotrophic mycena rosea – can be characterized mainly by a high abundance of mycorrhizal species, such as lactarius deliciosus, paxillus involutus and amanita pantherina. all the rare species of the core area are xylophags. to these belong ganoderma pfeifferi, hericium coralloides, hypsizygus ulmarius, flammulaster muricatus, ischnoderma resinosum, laxitextum bicolor, pluteus leoninus and p. umbrosus. the managed forest is poor in rare species. agaricus semotus and coprinus alopecia can be mentioned as examples. rare species of the plantation, like amanita pachyvolvata, cortinarius pseudovariicolor and lactarius subumbonatus, are all mycorrhizal. mycological monitoring 43 comparison of monitored areas based on species and fruit body numbers though the sizes of the monitored forests were distinct, difference among species richnesses of core areas, managed forests and plantations seemed to be significant. 224 species were found in the 600 000 m² core area, 88 species in the 3 160 000 m² managed forest and 48 species in the 15 000 m² plantation in the bükk mountains. table 2 demonstrates the numbers of registered species and fruit bodies in the 500 m² sampling plots in years 2001–2003. in respect of both species and fruit body numbers the core areas of the two monitored locations proved to be the richest ones. the species numbers of the core areas were nearly twice as much as those of the managed and the planted forests. in regard to the fruit body numbers a difference of one order of magnitude can be observed among fungal communities of the core areas, managed forests and plantations. ta b l e 2 cumulative numbers of species and fruit bodies of the macrofungal communities in the 500 m2 sampling plots in years 2001–2003 sampling plot number of species number of fruit bodies b1 149 33 103 b2 78 3350 b3 66 3163 m1 67 8185 m2 38 1898 m3 40 552 explanations: b1: core area, bükk mts b2: managed forest, bükk mts b3: plantation, bükk mts m1: core area, mecsek mts m2: managed forest, mecsek mts m3: plantation, mecsek mts. when the two locations are compared, it can be established that the sampling areas in the bükk mountains are richer regarding species and fruit body numbers. this fact can be explained first of all with a drier climate in the mecsek mountains. in addition, the shallower, rocky soil of the core area in the last one, the different ages of the two forests (őserdő in the bükk mountains is about 150–200 years old, whereas the reserve in the mecsek mountains is about 120–160 years old), and the substantial difference in the duration without any disturbance might have contributed to differences. functional characterization of fungal communities composition, structure and age of vegetation in addition to environmental conditions have a strong effect on the composition, species supply and quantitative relations of the fungal community developing in a site. thus functional spectra of the fungal communities indicate characteristics of the vegetation and its other environmental conditions. mycological monitoring 45 a relatively high ratio of mycorrhizal species in the plantation in the bükk mountains does not coincide with a higher number of mycorrhizal species as well; it is much more a consequence of driving back of saprotrophs (sh) and necrotrophic parasites (pn) by management. nevertheless, concerning the mecsek mountains, in the background of the higher ratio of mycorrhizal species in the managed forest and the plantation, there is a significant increase in species numbers, too. in most cases the proportion of soil-inhabiting saprotrophs (st) is higher in the managed forests and the plantations than in the core areas. this is, however, resulted in by the low number of lignicolous species, whereas the species number of saprotrophs is decreased. this suggests that in comparison with the managed forests and the plantations, the reserves provide more favorable conditions even for the soil-inhabiting saprotrophic communities. whereas the functional spectra of the two forest reserves seem to be fairly similar, this same is not true for the two managed forests, and is even less true for the two plantations. this, in addition to the different tree species compositions and mycorrhizal partners present, might probably be the consequence of the different soils, ages and managements of the forests. fig. 2. life form distribution (in %) of the sampling plots and in the aggregated samples (σ) in the bükk and the mecsek mountains based on the separate and the summarized data of the years 2001–2003. explanations: sh = lignicolous species, pn = necrotrophic parasites, m = mycorrhizal species, st = terricolous saprotrophs; b = bükk mts, m = mecsek mts; 1 = core area, 2 = managed forest, 3 = plantation. 46 f. pál-fám et al. figure 2 shows the functional spectra of the fungal communities of the plots in the different years. it seems that in both locations the spectra of the different years show higher fluctuation in the managed forests and the plantations than in the forest reserves. this suggests, that the core area provides more balanced conditions for the fungal communities, than the other two sites. in the case of the bükk mountains the spectra of the years 2001 and 2002 show high similarity, whereas in 2003 conspicuous is the increased dominancy of lignicoles (sh). though according to our knowledge the functional spectrum is a stable characteristic of the fungal community of a given habitat, this result indicates that in a dry period (like the year 2003), which is not favorable for developing fruit bodies, the spectrum can be changed towards a more definite dominance of lignicolous species. this can be explained by the fact, that wood retains humidity better than soil. in the case of mecsek, which has drier climate than the bükk montains, drought in the year of 2003 did not result in similar changes. analysis based on fruit body number studies of fungal communities by rank-abundance curves help to analyze their structure. the method is mostly used for the analysis of the structure of plant communities. communities being in an early stage of development are mainly characterized by a geometric series, whereas communities, which are rich in species, have more or less log-normal or “broken stick” distribution (m a g u r r a n 1988). �hereas the latter type shows a balanced distribution of abundances, the geometrical series indicates small number of species and an extremely unequal distribution of abundances within the community. according to our knowledge, this method has not been applied for fungal communities yet. diversity profiles give further informations about the structure of the studied community, which can not be acquired from rank-abundance curves. at 0 scale parameter the values shown by the curves depend on the number of species, whereas at bigger values of scale parameters the shape of the curve is influenced in a growing extent by the presence of dominant species. because of the different behaviour of species inhabiting trees (sh, pn) and soils (m, st), these groups were considered during our analyses as different communities. characterization of lignicolous, wood-inhabiting fungal communities rank-abundance curves of the lignicolous fungal communities (which include species of the groups sh and pn) of the different sampling sites show that the core areas have the highest species numbers and the most balanced distribution of species abundances (figs 3 and 4). the fewer species of the fungal communities of the managed forests, however, have a rather unequal distribution. consequently, the ability of the old, unmanaged forests, to preserve a great number of species, is better, than that of the managed forests, presumably because the first ones are able to provide more structured habitats. on the contrary, there is only a limited amount of dead wood in the managed forests, because actually all the timber cut down is transported away for utilization, and thus these forests provide less substrates – presumably also with lower variability and quality – for fungi. the smallest number and the most uneven distribution of lignicolous species can be detected in the plantations. these 52 f. pál-fám et al. fig. 11. ordination (pcoa analysis based on the coeffi cient of �accard) of the lignicolous fun-pcoa analysis based on the coefficient of �accard) of the lignicolous fun-) of the lignicolous fungal communities of the sampling sites of the bükk and mecsek mts. explanations: 1 = core area (b1), 2 = managed forest (b2), 3 = plantation (b3) in the bükk mts; 4 = core area (m1), 5 = managed forest (m2), 6 = plantation (m3) in the mecsek mts; variance: axis 1: 29%, axis 2: 24%. fig. 12. ordination (pcoa analysis based on the coefficient of �accard) of the soil-inhabiting fungal communities of the sampling sites of the bükk and mecsek mts. explanations: 1 = core area (b1), 2 = managed forest (b2), 3 = plantation (b3) in the bükk mts; 4 = core area (m1), 5 = managed forest (m2), 6 = plantation (m3) in the mecsek mts; variance: axis 1: 27%, axis 2: 21%. mycological monitoring 53 communities in the bükk mountains are more similar to each other, than those of the mecsek mountains. the significant separation of the plantation of the mecsek mountains from the other sampling sites of the same mountains is probably due to its lower species number and the bigger geographical distance from the other sampling sites. principal coordinate analysis of the data representing soil-inhabiting fungal communities gave a similar result (fig. 12). most significant is the difference between the two locations and the samples from the bükk mountains are less scattered than those from the mecsek mountains. the core area of the forest reserve in the mecsek mountains is remarkably separated from all other samples, suggesting that the species compositions of the investigated managed forest and plantation of the mecsek mountains are closer to each other, than to the similar feature of the core area. this supports the idea about the indicator values of fungi, that is, they indicate the less favourable characteristics of the shallow and rocky soil of the core area, as compared to the soils of the managed forest and the plantation. similarly to previous experiences (f o d o r 2003; s i l l e r et al. 2004), this monitoring confirms that the composition of fungal communities is strongly influenced by the geographical situation, management practices and other environmental conditions of habitats. our results suggest, that the significantly different species compositions of the different forest types existing in the proximity of each other make up together extraordinarily big species richness. this statement is supported by the results of our diversity studies, too. evaluation of the monitored areas from the point of view of conservation substantial part of species found in the monitored areas is also included in the recommended red list of macrofungi in hungary (r i m ó c z i et al. 1999). in the red list species are assigned to different categories on the basis of their endangerment following the recommendations of iucn (1994). category 1 includes the most endangered species, whereas category 4 includes those, which are also endangered, but occur more frequently. among species assigned to categories 1 and 2 (most endangered) 12 were found in our samples from the bükk and the mecsek mountains. on the basis of the number of endangered species, the prominent conservation value of the two core areas is clear (tab. 4). species falling into iucn category 2 can be practically found only here. these species are missing from the managed forests and the plantations monitored simultaneously, what refers to the fact, that rare species with special ecological demands find their life conditions only in undisturbed, unmanaged, natural or seminatural forests. lignicolous fungi demand dead wood material of different sizes and in different phases of decay, as substrate. species listed in category 3 of iucn occurred also in the plantations in big quantities. this can be explained by the fact, that many species of these plantations are connected to conifers, habitats of which are not indigenous in the monitored areas of hungary. 54 f. pál-fám et al. ta b l e 4 numbers and iucn categories of endangerment of species that are listed in the draft hungarian red list of macrofungi, found in the sampling sites sampling site iucn 1–2 iucn 3 iucn 4 b1 10 38 15 b2 1 21 3 b3 1 29 1 m1 6 17 15 m2 0 6 1 m3 0 24 2 explanations: b1: core area, bükk mts. b2: managed forest, bükk mts. b3: plantation, bükk mts. m1: core area, mecsek mts. m2: managed forest, mecsek mts. m3: plantation, mecsek mts. conclusions on the basis of the results of our monitoring activities in selected core areas of forest reserves, managed forests and plantations in the bükk and the mecsek mountains in hungary it can be established that the core areas are extremely rich in both species and fruit body numbers, thus being qualified as the best habitats in regard to wood-inhabiting fungi. on the basis of the presence of endangered species the core areas seem to be also important habitats for the species of special ecological demands. among fungal communities of the two locations those ones of the bükk mountains are richer in species, due to the older age of their forests and their more humid, cooler climate. perhaps forests of the bükk mountains are more sensitive to drought (see fig. 2), than the submediterranean type forests of the mecsek mountains. functional spectra of macrofungal communities characterized well the monitored sampling plots and their environmental conditions. since in years of extreme weather conditions the appearance of fruit bodies does not seem to represent the real species composition of the forests, it seemed reasonable to continue monitoring in the course of at least 3 years. most endangered species are necrotrophic parasites (pn), which indicate impacts of human management with drastic decreases in fruit body numbers, or by complete disappearance. the other endangered group includes the wood-inhabiting saprotrophs (sh), from which many species disappear as a consequence of anthropogenic effects. in both investigated locations the most structured lignicolous fungal communities can be found in the core areas of forest reserves: these are characterized by the highest number of species, with some dominant, several moderately rare and even more rare species. the least structured are the lignicolous assemblages of the plantations – here even the presence of any community organization can not be confirmed. in respect to rare species, the core areas are the richest in both locations, followed by the managed forests and finally by the plantations. in supporting the survival of the lignicolous fungi and the maintenance of the community structure the quantity and the composition of dead wood material in the forest reserves is very important. the diversity of dead wood material, serving as nutrient for different fungi, and the different stages of the decomposition process increase biodiversity significantly. mycological monitoring 55 independently of the spatial position, the core areas have a lignicolous fungal community of typical functional composition and structure, which developed in a similar way in both locations (see figs 1–6 and 11). consequently, it can be presumed that the geographical position does not have a significant impact upon the functional composition and the structure of the fungal communities of forest reserves. in the case of the managed forests and the plantations, however, presumably as a consequence of the limited availability of dead wood, the geographical location and the environmental factors are more important determining factors of the forest structure and functional composition, than the degree of naturalness of forests. dead wood seems to influence the functional composition and the structure of fungal communities in a similar way as rivers influence the species composition and the structure of azonal plant communities. soil-inhabiting fungal communities of the managed forests and the plantations show a relatively varied, diverse structure in both locations. it is surprising, that – in contrast with the lignicolous fungal communities – the soil-inhabiting fungal communities of the core areas have less species and a more simple structure than the managed forests and the plantations. nevertheless, the species compositions of the soil-inhabiting fungal communities of the two locations are different. presumably the appearance of fungal species connected to the special tree species of the managed forests and the plantations makes these communities more diverse. this higher diversity, however, is not accompanied by an increased number of species of high nature conservation value. on the basis of the results presented it can be concluded, that the composition of the soil-inhabiting fungal communities is primarily influenced by the composition of the vegetation and the type of soil, whereas the state of naturalness of the forest and the type of its management are less important determining factors. in the separate years only a part of the fungal species present in the habitats produces fruit bodies. consequently, in order to make the results of a mycological monitoring representative of the habitats, collection of data during several years is necessary. if also the results of our previous surveys are taken into consideration, the continuation of monitoring for at least 3 years seems to be required. anyhow, for the comparison of functional groups, diversities and species compositions of fungal communities, monitoring of 3 years seems to be enough. the present mycological monitoring confirmed that forest reserves are very important as refugia for fungi, and offer a possibility for them to spread later into the area of presently managed forests, too. during our monitoring some species were found, which are good indicators of certain characteristics (e.g. degree of naturalness, age, availability of dead wood and completeness of the decay cycle) of forests. our results support that the protocol elaborated in the frame of hbms is suitable for a mycologically based comparison of different forests, furthermore for the evaluation of the state of forests. additionally, long-term monitoring can help in the recognition and description of forest development processes, changes and trends. importances of long-term monitoring of fungal communities are as follows: 1) basic data are collected about changes in the species and functional composition of the fungal communities, 2) standardized sampling offers an opportunity for comprehensive comparisons as well as observations of dynamics and trends of scientific demand, 56 f. pál-fám et al. 3) on the basis of long-term monitoring of fungal communities in different geographical regions and in forests under different management practices, additional indicator species can be identified, 4) within the same sampling locations significance can be demonstrated between the results of fungal monitoring and some other components monitored (e.g. characteristics of the vascular vegetation or the abiotic environment); as a consequence, indicative characteristics of fungi can be used in the frame of comlex studies. 5) mycological monitoring confirms that forest reserves are very important as refugia for fungi, and offer a possibility for them to spread later into the area of presently managed forests, too, 6) it makes possible a better estimation of the threatened status of species. acknowledgements: our monitoring activity was supported by the hungarian ministry of environment and water in the framework of the hungarian biodiversity monitoring system and the faculty of veterinary sciences of the szent istván university (nkb-2002-kut-6). we are 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(in:) f. h o r v á t h , a. b o r h i d i (eds). aims, conceptions and methods of forest reserve investigations. természetbúvár alapítvány kiadó, budapest. 182–202. 58 f. pál-fám et al. s i l l e r i., p á l f á m f., f o d o r l. 2004. erdők állapotváltozásának nyomon követése nagygombák segítségével. (macrofungi as indicators of forest regeneration and forest developmental processes.) természetvédelmi közlemények 11: 185–194. s k i r g i e ł ł o a. 1998. macromycetes of oak-hornbeam forests in the bialowieza national park – monitoring studies. acta mycologica 33 (2): 171–189. s o d e r m a n g. (ed.) 1998. master plan for monitoring biodiversity in estonia. environmental information centre, tallin. phare s t o k l a n d �. n. 2001. the coarse woody debris profile: an archive of recent forest history and an important biodiversity indicator. ecological bulletins 49: 71–84. t ó t h m é r é s z b. 1993. nucosa 1.0: number cruncher for community studies and other ecological applications. abstracta botanica 7: 283–287. t ó t h m é r é s z b. 1997. diverzitási rendezések. (diversity orderings.) scientia, budapest. tr e c z k e r k., s z a b ó i. 2002. farontó gombák a ropolyi erdőrezervátumban. (�ood decay fungi from the forest reserve of ropoly.) clusiana mikológiai közlemények 41 (2–3): 67–94. ve e r k a m p m., k u y p e r t. w. 1993. mycological investigations in forest reserves in the netherlands. (in:) m. e. a.b r o e k m e y e r , w. vo s , h. k o o p (eds). european forest reserves. proceedings of the european forest reserves workshop, 6–8 may 1992, wageningen: 127–143. w h i t t a k e r r. h. 1970. communities and ecosystems. 1st ed. macmillan, new york. w i n t e r h o f f �. 1989. die bedeutung der baden-würtenbergischen bannwälder für den pilzartenschutz. waldschutzgebiete in rahmen der mitteilungen der forstlichen versuchsund forschungsanstalt. band 4: 183–190. w o j e w o d a w., h e i n r i c h z., k o m o r o w s k a h. 1999. macromycetes of oak-lime-hornbeam woods in the niepołomice forest near kraków (s poland) – monitoring studies. acta mycologica 34 (2): 201–266. 2014-01-01t11:45:06+0100 polish botanical society fungal biodiversity in rhizosphere of healthy and needle castaffected scots pine transplants małgorzata mańka department of forest pathology, the august cieszkowski agricultural university wojska polskiego 71c, pl-60-625 poznań, mmanka@au.poznan.pl m a ń k a m.: fungal biodiversity in rhizosphere of healthy and needle cast-affected scots pine transplants. acta mycol. 42 (2): 199-202, 2007. healthy scots pine (pinus sylvestris l.) transplants had in rhizosphere a community of saprotrophic fungi which considerably suppressed the growth of severe root pathogens heterobasidion annosum and armillaria obscura. a community from transplants affected by needle cast (lophodermium spp.) suppressed both pathogens to a much smaller extent. key words: scots pine, rhizosphere fungi, needle-cast, heterobasidion, armillaria introduction fungi in the soil environment of host-plant may reflect the probable effect of the environment they derive from, on a pathogen’s activity towards its host-plant (m a ń k a k. 1990). communities of fungi from rhizosphere of scots pine (pinus sylvestris l.) transplants can effect growth of root pathogenic fungi in various ways, depending on year, season of the year and transplant’s age (m a ń k a k. et al. 1993; m a ń k a m. 1995b). the aim of the work was to investigate the influence of needle cast attack on scots pine transplants rhizosphere fungi communities, and on the communities’ effect on growth of severe root pathogens heterobasidion annosum (fr.) bref. and armillaria obscura (schaeff.) herink. materials and methods in konstantynowo forest nursery (near poznań) 2-year-old scots pine (p. sylvestris) transplants (sown in april 1992, on deep sandy forest soil) grew in two groups: well developed healthy ones, and poorly developed needle cast-affected (lophodermium spp.) ones. on april 18, 1994, rhizosphere soil from both groups was sampled by shaking off (m a ń k a k., m a ń k a m. 1993) and communities of soil fungi were isolated according to warcup’s soil plate method modified by k. mańka (wa r c u p acta mycologica vol. 42 (2): 199-202 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 200 m. mańka 1950; m a ń k a k. 1964; j o h n s o n , m a ń k a k. 1961; m a ń k a k., s a l m a n o w i c z 1987). the 2 communities (with specific structure and function) were considered characteristic of the rhizosphere soils investigated. both communities were tested for their effect on growth of h. annosum type p and a. obscura with the biotic series method by k. mańka (m a ń k a k. 1974; m a ń k a k., m a ń k a m. 1992; m a ń k a m. 1995a). the test was performed on pda for h. annosum (biotic effect estimated after 10 days) and on malt agar for a. obscura (biotic effect estimated after 20 days). the biotic test resulted in describing the phytopathological function of both communities. the function is expressed by summary biotic effect (sbe), i.e. the effect of the entire soil fungi community on the pathogen’s growth. the sbe results from summarizing all the general biotic effects (gbe = an effect of all the isolates of a species on the pathogen’s growth). a gbe results from multiplying frequency of the species by individual biotic effect (ibe) value, that is the effect of one isolate of the given species on the pathogen’s growth. any of the biotic effects may be positive (indicating suppressive effect on the pathogen’s growth), negative (indicating supporting effect on the pathogen’s growth) or neutral (“0”). intensity of the supporting/suppressing effect is described by the absolute value of the effect. results and discussion the fungal community from healthy transplants was more numerous (by 28%, tabs 1 and 2) than the community from the needle cast-affected ones. the species composition was not very different. the most remarkable difference was to be seen in trichoderma spp. – there were all together 41 isolates of t. koningii and t. longibrachiatum in the community from healthy transplants, and only 8 isolates of t. viride in the community from diseased ones (tabs 1 and 2). an interesting phenomenon was observed: representatives of the same species happened to behave variously, with respect to a pathogen, depending on from what community they derived. their individual biotic effect (ibe) supported the pathogen to a greater extent, when they were isolated from diseased plant rhizosphere. it was most evident in coniothyrium fuckelii, which had a very suppressing effect on h. annosum when isolated from healthy pine rhizosphere (ibe +10, tab. 1), and a slightly supporting effect when isolated from diseased pine rhisosphere (ibe -2, tab. 2). the same was true for umbelopsis vinacea with ibe -2 and -5, and fusarium oxysporum with ibe +3 and 0, when derived from healthy and diseased plants, respectively. all the summary biotic effects (sbe) exerted by the rhizosphere fungi communities on both pathogens were positive, which means that the growth of both pathogens was suppressed by both communities. the suppression of h. annosum was much bigger in rhizosphere of healthy pine seedlings (sbe +1342, tab. 1) than of diseased seedlings (sbe +236, tab. 2). the situation was similar with respect to a. obscura – the rhizosphere fungi community from healthy pine seedlings had sbe +721 (tab. 1), and from diseased ones – sbe +443 (tab. 2). this may mean that the needle cast contributed not only to worse development of scots pine transplants but also to formation in their rhizosphere of a fungal community that could suppress both pathogens in question to a fungal biodiversity 201 smaller extent, than the fungal community in the rhizosphere of healthy transplants. it seems that the needle cast, being a disease of the over ground part of the plant, may considerably influence biotic relations in soil. ta b l e 1 biotic effect of rhizosphere fungi communities from healthy 2-year-old scots pine transplants on the growth of heterobasidion annosum and armillaria obscura species of fungi fre-quency biotic effects on heterobasidion annosum armillaria obscura ibe gbe ibe gbe umbelopsis vinacea (dixon-stew.) arx 59 -2 -118 +2 +118 paecilomyces marquandii (masee) hug. 52 +6 +312 +1 +52 coniothyrium fuckelii sacc. 48 +10 +480 0 0 trichoderma koningii oudem. 36 +8 +288 +8 +288 penicillium daleae zaleski 26 +3 +78 +1 +26 penicillium sp. 1 21 +6 +126 +4 +84 truncatella truncata (lév.) steyaert 15 0 0 +4 +60 penicillium steckii zaleski 7 +13 +91 0 0 fusarium oxysporum schlecht. 7 +3 +21 +4 +28 trichoderma longibrachiatum rifai 5 +8 +40 +8 +40 chaetomium aureum chivers 5 +3 +15 +3 +15 penicillium brevicompactum dierckx 3 +1 +3 0 0 dematiaceae sp. 1 2 -4 -8 +1 +2 zygorhynchus moellerii vuill. 2 +7 +14 +4 +8 summary biotic effect 288 +1342 +721 explanations: ibe individual biotic effect; gbe general biotic effect; sbe summary biotic effect. ta b l e 2 biotic effect of rhizosphere fungi communities from needle cast-affected 2-year-old scots pine transplants on the growth of heterobasidion annosum and armillaria obscura species of fungi fre-quency biotic effects on heterobasidion annosum armillaria obscura ibe gbe ibe gbe coniothyrium fuckelii sacc. 55 -2 -110 0 0 penicillium sp. 2 34 +12 +408 -1 -34 umbelopsis vinacea (dixon-stew.) arx 33 -5 -165 0 0 penicillium sp. 3 31 0 0 +6 +186 chaetomium aureum chivers 17 0 0 +4 +68 truncatella truncata (lév.) steyaert 15 -1 -15 +4 +60 penicillium nigricans bainier ex thom 9 +4 +36 +5 +45 chaetomium globosum kunze ex steud. 8 +2 +16 0 0 trichoderma viride pers. ex gray 8 +8 +64 +7 +56 penicillium sp. 4 5 -5 -25 +6 +30 fusarium oxysporum schlecht. 5 0 0 +4 +20 zygorhynchus moellerii vuill. 3 +7 +21 +5 +15 penicillium verruculosum dierckx 2 +3 +6 -1 -3 summary biotic effect 225 +236 +443 legend see tab. 1 202 m. mańka conclusion the scots pine transplants affected by needle cast should not be used for establishing plantations not only because of their poor development and poor growth prospects (which is obvious), but also because they are not able to resist soil-borne pathogenic fungi to an extent comparable to healthy transplants. references j o h n s o n l. f., m a ń k a k. 1961. a modification of warcup’s soil plate method for isolating soil fungi. soil science 2: 79–83. m a ń k a k. 1964. próby dalszego udoskonalenia zmodyfikowanej metody warcupa izolowania grzybów z gleby. ptpn, prace kom. nauk rol. i kom. nauk leś. 17: 29–43. m a ń k a k. 1974. zbiorowiska grzybów jako kryterium oceny wpływu środowiska na choroby roślin. zesz. probl. post. nauk rol. 160: 9–23. m a ń k a k. 1990. saprofityczna mikoflora środowiska glebowego a zdrowotność roślin. phytopathol. pol. 11: 122–133. m a ń k a k., s a l m a n o w i c z b. 1987. udoskonalenie niektórych technik zmodyfikowanej metody płytek glebowych do izolowania grzybów z gleby z punktu widzenia potrzeb mikologii fitopatologicznej. rocz. nauk rol., seria e 17 (1): 35–46. m a ń k a k., m a ń k a m. 1992. a new method for evaluating interaction between soil inhabiting fungi and plant pathogens. iobc/wprs bulletin 15 (1): 73–75. m a ń k a k., m a ń k a m. 1993. o metodzie izolacji grzybów z ryzosfery drzew leśnych. materiały z iv konferencji sekcji biologicznych metod ochrony roślin przed chorobami, 22-23 04 1993, ptfit., skierniewice: 3–7. m a ń k a k., m a ń k a m., k w a ś n a h., ł a k o m y p., b a b k i e w i c z m. 1993. zagrożenie sadzonek drzew leśnych przez patogeny korzeni a zbiorowiska grzybów ryzosferowych. materiały z iv konferencji sekcji biologicznych metod ochrony roślin przed chorobami, 22-23.04.1993, ptfit., skierniewice: 7–13. m a ń k a m. 1995 a. non-pathogenic soil fungi reflecting soil environment. (in:) m. m a ń k a (ed.). environmental biotic factors in integrated plant disease control, proceedings of 3rd conference of european foundation for plant pathology, september 5-9, 1994, poznań: 27–36. m a ń k a m. 1995 b. effect of rhizosphere fungi communities from scots pine and birch transplants on growth of root pathogenic fungi. (in:) l.b. o r l i k o w s k i , cz. s k r z y p c z a k (eds). biological control of soil-borne and postharvest pathogens. the polish phytopathological society, section of biological control of plant diseases, research institute of pomology and floriculture, skierniewice: 61–64. wa r c u p j. h. 1950. the soil plate method for isolation of fungi from soil. nature 166: 117–118. bioróżnorodność grzybów z ryzosfery zdrowych i porażonych przez osutkę sadzonek sosny zwyczajnej s t r e s z c z e n i e porównano zbiorowiska grzybów ryzosferowych dwuletnich sadzonek sosny zwyczajnej: zdrowych i dorodnych oraz niewyrośniętych, porażonych przez osutkę (lophodermium spp.). zbiorowisko grzybów ze zdrowych sadzonek sosny ograniczało wzrost patogenów korzeni heterobasidion annosum i armillaria obscura znaczniej bardziej niż zbiorowisko z porażonych przez osutkę sadzonek. sadzonki porażone przez osutkę nie nadają się do wysadzania na uprawy leśne nie tylko ze względu na zahamowanie wzrostu i ograniczony przez chorobę aparat asymilacyjny, ale również z powodu tego, że ich zbiorowiska grzybów ryzosferowych nie są w stanie ograniczyć wzrostu patogenów obecnych w glebie leśnej w takim stopniu, jak zbiorowiska ryzosferowe zdrowych sadzonek. 2014-01-01t11:46:14+0100 polish botanical society japewia tornoensis and further localities of j. subaurifera found in the carpathians paweł czarnota department of agroecology, university of rzeszów, ćwiklińskiej 2, pl-35-601 rzeszów scientific laboratory of the gorce national park, poręba wielka 590, pl-34-735 niedźwiedź pawczarnota@poczta.onet.pl czarnota p.: japewia tornoensis and further localities of j. subaurifera found in the carpathians. acta mycol. 44 (2): 259–264, 2009. japewia tornoensis is reported for the first time from the carpathians and poland. further localities of j. subaurifera, known so far from a single carpathian collection in the polish tatra mts., are also presented. some diagnostic features and general distribution of both species are provided and similar taxa are discussed. key words: japewia, lichenized fungi, sorediate lichens, western carpathians, poland introduction the lichen forming genus japewia (lecanorales, ascomycota), described recently by tønsberg (1990) includes only two species, j. subaurifera muhr & tønsberg and j. tornoensis (nyl.) tønsberg. from poland the first one was reported so far. its single locality in the polish tatra mountains was also known as only one in the whole carpathians (bielczyk et al. 2004; czarnota, kukwa 2004). the author supposed, however, that the species also occurs in other parts of this central european mountains, at least in regions covered with upper montane spruce forests. field investigations carried out in western carpathians in 2008–2009 confirmed that suggestion and brought several succeeding records of j. subaurifera. surprisingly, during this intensive work on diversity of epixylic lichens presuming dead spruces, j. tornoensis has also been discovered in the gorce mts. being a new lichen species for the carpathians and poland. acta mycologica vol. 44 (2): 259–264 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 260 p. czarnota material and methods the material was examined with light microscopes. hand-made apothecial sections and squashed thallus preparations, were studied in water and potassium hydroxide solution (k). collections of j. subaurifera were compared with other polish materials checked previously by b. j. coppins (uk). the material is stored in the herbarium of the gorce national park (gpn). the nomenclature follows index fungorum (http://indexfungorum.org; date of exploration: july 2009) and liška et al. (2008). the species japewia subaurifera muhr & tønsberg, lichenologist 22 (3): 206 (1990). morphology and affinities. descriptions and illustrations of the species are given for example by tønsberg (1990) and ohmura & kashiwadani (1997). all collections presented here correspond well with those descriptions and tatra’s specimens determined previously by coppins (czarnota, kukwa 2004). they are only sterile as most european findings forming brown-yellow, orange-brown to yellowish green soralia (fig. 1). because of similarly coloured soralia j. subaurifera resembles lecidea pullata (norman) th. fr. and caloplaca lucifuga thor. it is especially similar to the first species which covers often the same substrates being common in upper montane belt in carpathians. the second one grows mainly on deciduous trees, particularly often in crevices of rough bark of old quercus spp., rather in lowland woodlands than in mountains. each of them differ in the chemistry since j. subaurifera produces lobaric acid and several ‘subaurifera’ pigments, l. pullata spaerophorin and isosphaeric acid, while c. lucifuga has parietin and fallacinal (tønsberg 1992; wirth 1995). from those reasons the orange-brown soralia of the last species react k+ red-violet in contrast to k+ fuscous brown soralia of j. subaurifera (visible in a squash preparation). soralia of l. pullata give no reaction with k or they are only k ± yellowish. ecology. the species grows on bark of several deciduous trees, mainly betula spp., alnus incana and sorbus aucuparia (tønsberg 1992) but except scandinavia it is found more often on different conifer trees in large woodlands from about see-level in north-west norway to upper montane belt in mountains of central europe and japan. in poland it was collected only in upper montane spruce forest plagiotheciopiceetum growing as an epiphyte on picea abies and pinus cembra or as an epixyle of hard wood on decorticated spruce trunks. it prefers old, natural stands showing some implication to be an indicator of ecological continuity of these forest ecosystems. its ecological role was taken into account also in scottish native pinewoods, where j. subaurifera is considered as the ‘bonus’ species (coppins, coppins 2002). accompanied taxa for the new polish findings include: calicium trabinellum (ach.) ach., chaenotheca spp., elixia flexella (ach.) lumbsch, fuscidea pusilla tønsberg, hypogymnia physodes (l.) nyl., hypocenomyce caradocensis (leight. ex nyl.) p. james & gotth. schneid., h. scalaris (ach.) m. choisy, lecanora conizaeoides nyl. ex cromb., l. phaeostigma (körb.) almb., l. pulicaris (pers.) ach., l. subintricata japewia tornoensis and further localities 261 (nyl.) th. fr., lecidea leprarioides tønsberg, l. nylanderi (anzi.) th. fr., l. turgidula fr. and mycoblastus sanguinarius (l.) norman. world distribution. j. subaurifera is a widespread lichenized fungus reported mainly from the zone of boreal forests in the northern hemisphere. it is especially frequent in scandinavia; particularly in norway and sweden (tønsberg 1990). from other european sites, including scotland, iceland, komi republic in russia, estonia and mountainous areas of portugal, the czech republic (šumava, české švýcarsko, eastern sudetes), austria and turkey it is known from sparse localities (boom van den, giralt 1996; kristinsson 1999; palice 1999; halonen et al. 2000; tønsberg et al. 2001; czarnota, kukwa 2004; halda 2006). j. subaurifera seems to be also widespread throughout boreal zone of north america (tønsberg 1990, 1992; mccune et al. 2000; hauck et al. 2006); its locally frequent occurrence on japan island hokkaido is also known (ohmura, kashiwadani 1997). up to date the single locality from polish tatra mts. was considered as only one in whole carpathians (czarnota, kukwa 2004). the new findings presented here extend its occurrence on other ranges of western carpathians. specimens examined (all within upper montane spruce forest plagiothecio-piceetum). poland. atpol grid square gd 27 − western beskidy mts., babia góra massif, babia góra national park, ne slope of sokolica mt., 49°35’10.4”n, 19°34’06.5”e, alt. 1250 m, on wood of decorticated trunk of picea abies, 8 june 2009, leg. p. czarnota 5976 (gpn) and 49°35’08.8”n, 19°34’07.9”e, alt. 1275 m, on bark of picea abies, 8 june 2009, leg. p. czarnota 5981 (gpn). ge 11 − western beskidy mts., gorce mts., gorce national park, forest section no. 8c, n slope of kudłoń mt., 49°34.448’n, 20°10.583’e, alt. 1210 m, on wood of picea abies, 18 may 2009, leg. p. czarnota 5986 (gpn). ge 21 − gorce mts., gorce national park, forest section no. 184b, w of gabrowska glade, 49°32.595’n, 20°08.510’e, alt. 1245 m, on bark of picea abies, 29 aug. 2008, leg. p. czarnota 5632 (gpn). japewia tornoensis (nyl.) tønsberg – lichenologist 22 (3): 206 (1990). syn. lecidea tornoënsis nyl., herb. mus. fenn.: 110 (1859); biatora tornoënsis (nyl.) th. fr., nova acta r. soc. scient. upsal. 3: 296 (1861); mycoblastus tornoënsis (nyl.) r. a. anderson, bryologist 77 (2): 220 (1974). morphology and affinities. considering similar lichens growing in the upper montane belts of western carpathians, j. tornoensis at first sight resembles paler (shaded) forms of strangospora moriformis (ach.) stein by semiglobose, immarginate, brown to dull brown apothecia (fig. 2). mature apothecia of lecanora phaeostigma with reduced excipulum are also very similar to those formed by j. tornoensis (fig. 3). growing closely to the both species, j. tornoensis could easily be overlooked thus its real distribution in the carpathians seems to be wider than we currently known. its distinguishing from the letter species is quite simple by anatomic characters, mainly by distinct differences in ascus shape and strongly thickened spores (walls 1.5–3µm), which are large (15–20 × 10–15µm) and additionally coated by thick gelatinous epispore (figs 4, 5). l. phaeostigma has much smaller, ellipsoid and thin-walled spores and moreover very often visible numerous brown black pycnidia. s. moriformis forms globose, small ascospores and multispored asci. for more details see for example tønsberg (1990) and hawksworth & coppins (1992). ecology. except antarctica and oceanic parts of europe, where j. tornoensis has been found on terricolous bryophytes and humus in rock crevices (hawksworth, coppins 1992; olech 2004), almost all its inland reports are made in subalpine belts covered with conifer forest. it was collected there mainly as an epiphyte on, e.g., 262 p. czarnota larix decidua, pseudotsuga manziesii, picea engelmannii, abies lasiocarpa and rarely on wood. studies of hauck and spribille (2005) in spruce-fir forests of northwestern montana suggest that j. tornoensis principally occupies branches of conifer trees. perhaps the species prefers the same kind of habitat also in the carpathians, as would indicate the finding presented here. world distribution. j. tornoensis is a widespread bipolar species, however except arctic regions is rather rarely reported elsewhere. it is known from austria (alps; e.g. obermayer 1997; hafellner et al. 2003), germany (bayern; scholz 2000), whole scandinavia (santesson 2004) including svalbard (elvebakk, hertel 1997), switzerland (scheidegger et al. 2002), british isles (c scotland; hawksworth, coppins 1992), iceland, greenland, canada, alaska, almost all geographical regions of arctic russia (thomson 1997; kristinsson et al. 2006), mongolia (hauck, javkhlan 2008), japan (kurokawa 2003), usa: e.g., colorado (anderson, carmer 1974), arizona (obermayer 1997), new york (harris 2004), montana (hauck, spribille 2005) as well as antarctica (øvstedal, lewis smith 2001; olech 2004). specimen examined. poland. ge 21 − western beskidy mts, gorce mts., gorce national park, forest section no. 158a, e of jaworzyna kamienicka mt., 49°32.794’n, 20°09.901’e, alt. 1250 m, on hard wood of spruce branch within upper montane spruce forest plagiothecio-piceetum, 2 oct. 2008, leg. p. czarnota 5637 (gpn), det. z. palice. acknowledgements. dr. z. palice (průhonice, czech republic) is thanked for turn my attention towards j. tornoensis and his hospitality during my stay in průhonice in 2009. the work is a part of project supported by polish ministry of science and higher education, grant no. n n304 306835. references anderson r. a., carmer m. b. 1974. additions to the lichen flora of colorado. bryologist 77: 216–223. bielczyk u., lackovičová a., farkas e., lőkös l., liška j., breuss o., kondratyuk s.y. 2004. checklist of lichens of the western carpathians. w. szafer institute of botany, polish academy of sciences, kraków, 181 pp. boom van den p. p. g., giralt m. 1996. contribution to the flora of portugal, lichens and lichenicolous fungi i. nova hedwigia 63 (1/2): 145–172. coppins a. m., coppins b. j. 2002. indices of ecological continuity for woodland epiphytic lichen habitats in the british isles. british lichen society, wimbledon, 36 pp. czarnota p., kukwa m. 2004. some sorediate lichens and lichenicolous fungi new to poland. graphis scripta 15: 24–32. elvebakk a., hertel h. 1997. a catalogue of svalbard lichens. 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(ed.). 2003. checklist of japanese lichens. national science museum, tokyo, 128 pp. liška j., palice z., slavíková š. 2008. checklist and red list of lichens of the czech republic. preslia 80: 151–182. mccune b., rosentreter r., ponzetti j. m., shaw d. c. 2000. epiphyte habitats in an old conifer forest in western washington, u.s.a. bryologist 103: 417–427. obermayer w. 1997. lichenotheca graecensis. fasc. 4 (nos 61–80). fritschiana 8: 1–6. ohmura y., kashiwadani h. 1997. lichens of mt. o-akan and its adjacent areas, hokkaido, japan. bulletin of the national science museum (tokyo), series b (botany) 23: 1–24. olech m. 2004. lichens of king george island, antarctica. the institute of botany of the jagiellonian university, kraków, 391 pp. øvstedal d. o., lewis smith r. i. 2001. lichens of antarctica and south georgia. cambridge university press, cambridge, 411 pp. palice z. 1999. new and noteworthy records of lichens in the czech republic. preslia 71: 289–336. santesson r., moberg r., nordin a., tønsberg t., vitikainen o. 2004. lichen-forming and lichenicolous fungi of fennoscandia. museum of evolution, uppsala university, uppsala, sweden, 359 pp. scheidegger c., clerc p., dietrich m., frei m., groner u., keller c., roth i., stofer s., vust m. 2002. rote liste der gefährdeten arten der schweiz: baumund erdbewohnende flechten. hrsg. bundesamt für umwelt, wald und landschaft buwal, bern and eidgenössische forschungsanstalt wsl, birmensdorf and conservatoire et jardin botaniques de la ville de genève cjbg. buwal-reihe vollzug umwelt, 124 pp. scholz p. 2000. katalog der flechten und flechtenbewohnenden pilze deutschlands. schriftenreihe für vegetationskunde 31: 4–298. thomson j. w. 1997. american arctic lichens. ii. the microlichens. university of wisconsin press, madison, 688 pp. tønsberg t. 1990. japewia subaurifera, a new lichen genus and species from north-west europe and western north america. lichenologist 22: 205–212. tønsberg t. 1992. the sorediate and isidiate, corticolous, crustose lichens in norway. sommerfeltia 14: 1–331. tønsberg t., türk r., hofmann p. 2001. notes on the lichen flora of tyrol (austria). nova hedwigia 72 (3/4): 487–497. wirth v. 1995. die flechten baden-württembergs. verlag e. ulmer, stuttgart, 1006 pp. 264 p. czarnota japewia tornoensis i kolejne stanowiska j. subaurifera odnalezione w karpatach streszczenie rodzaj japewia (grzyby zlichenizowane) obejmuje dwa gatunki, j. tornoensis i j. subaurifera. pierwszy, o bipolarnym charakterze występowania na świecie, został stwierdzony po raz pierwszy w karpatach (gorce) i w polsce. drugi, znany przede wszystkim z borealnych obszarów półkuli północnej, notowany był dotychczas w karpatach tylko na jednym stanowisku, w tatrach (czarnota, kukwa 2004). teraz odnaleziono go także w gorcach i na babiej górze. oba gatunki w całej europie środkowej są uznawane za rzadkie. j. subaurifera występuje przede wszystkim w postaci płonnej, wykształcając charakterystyczne, brunatnawożółte, pomarańczowobrunatne lub zielonawożółte soralia produkujące kwas lobariowy i kilka pigmentów „subaurifera” (m. in. secalonic acid i eumitryn). w karpatach zachodnich gatunek ten rośnie na korze starych świerków i na twardym drewnie pni świerkowych w górnoreglowym borze świerkowym plagiothecio-piceetum. lokuje się ponad podstawą pnia, zajmując tym samym odmienną niszę ekologiczną niż lecidea pullata – inny, morfologicznie podobny, „sorediowany” i częsty w tym zbiorowisku leśnym grzyb zlichenizowany. j. tornoensis wykształca półkoliste, brunatnoczerwone apotecja przypominające wyglądem jaśniejsze (ocienione) formy strangospora moriformis, a także scoliciosporum chlorococcum czy też lecanora phaeostigma. gatunek ten łatwo jest odróżnić po charakterystycznych, grubościennych (1,5–3μm), jednokomórkowych, dużych zarodnikach (15–20 x 10–15μm). podobieństwo do tych pospolitych porostów pozwala przypuszczać, że może to być gatunek częstszy w karpatach, lecz przeoczany. figs 1-2. habits of japewia: 1. j. subaurifera (czarnota, gpn 5981), 2. j. tornoensis (czarnota, gpn 5637). figs 3-5. japewia tornoensis (czarnota, gpn 5637): 3. apothecial section. scale bar = 100 μm, 4-5. ascospores with strongly thickened walls. scale bars = 10 μm. 2014-01-01t11:49:49+0100 polish botanical society water-borne hyphomycetes in tree canopies of kaiga (western ghats), india naga m. sudheep and kandikere r. sridhar department of biosciences, mangalore university mangalagangotri, mangalore in-574-199, india, sirikr@yahoo.com sudheep n. m., sridhar k. r.: water-borne hyphomycetes in tree canopies of kaiga (western ghats), india. acta mycol. 45 (2): 185–195, 2010. the canopy samples such as trapped leaf litter, trapped sediment (during summer), stemflow and throughfall (during monsoon) from five common riparian tree species (artocarpus heterophyllus, cassia fistula, ficus recemosa, syzygium caryophyllatum and xylia xylocarpa) in kaiga forest stand of the western ghats of southwest india were evaluated for the occurrence of water-borne hyphomycetes. partially decomposed trapped leaf litter was incubated in bubble chambers followed by filtration to assess conidial output. sediments accumulated in tree holes or junction of branches were shaken with sterile leaf disks in distilled water followed by incubation of leaf disks in bubble chamber and filtration to find out colonized fungi. stemflow and throughfall samples were filtered directly to collect free conidia. from five canopy niches, a total of 29 water-borne hyphomycetes were recovered. the species richness was higher in stemflow and throughfall than trapped leaf litter and sediments (14-16 vs. 6-10 species). although sediments of syzygium caryophyllatum were acidic (5.1), the conidial output was higher than other tree species. stemflow and throughfall of xylea xylocarpa even though alkaline (8.5-8.7) showed higher species richness (6-12 species) as well as conidial load than rest of the tree species. flagellospora curvula and triscelophorus acuminatus were common in trapped leaf litter and sediments respectively, while conidia of anguillospora crassa and a. longissima were frequent in stemflow and throughfall. diversity of water-borne hyphomycetes was highest in throughfall of xylea xylocarpa followed by throughfall of ficus recemosa. our study reconfirms the occurrence and survival of diverse water-borne hyphomycetes in different niches of riparian tree canopies of the western ghats during wet and dry regimes and predicts their possible role in canopy as saprophytes, endophytes and alternation of life cycle between canopy and aquatic habitats. key words: water-borne hyphomycetes, diversity, conidia, canopy, leaf litter, sediment, stemflow, throughfall acta mycologica vol. 45 (2): 185–195 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 186 n. m. sudheep and k. r. sridhar introduction forest canopies are endowed with a mosaic of flora, fauna and microbes (nadkarni et al. 2001). tropical canopy of ecuador trap about 200-300 kg dry mass of litter per hectare consisting of at least 500-600 m of rhizomorphs (e.g., marasmius spp.) per kg litter, which is equivalent to approximately 180 km per hectare (hedger 1990). a rough estimate of microfungal biomass in twigs and needle surfaces of old-growth douglas fir forest canopies was up to 450 kg/ha/yr (carroll et al. 1980). thus, canopies provide a wide variety of niches and accommodate a broad group of fungi (e.g., phylloplane fungi, endophytes, pathogens, lignicolous fungi) (lodge, cantrell 1995; stone et al. 1996; shaw 2004). besides these, water-borne fungi (aquatic and aero-aquatic hyphomycetes) are also known from canopies. they have been reported from the epiphytes, tree holes, trapped leaf litter, stemflow, throughfall and other niches of canopy in temperate and tropical regions (e.g., ando, tubaki 1984; czeczuga, orłowska 1998a, 1999; gönczöl, révay 2006; sridhar et al. 2006; karamchand, sridhar 2008, sridhar 2009; sridhar, karamchand 2009). typical water-borne hyphomycetes are also endophytic in needles of black spruce (picea mariana) of a mixed woody forest in canada (sokolski et al. 2006). tree holes (formed due to decomposition of branches) and junctions of branches act as receptacles for deposition of water, sediment, leaf litter and other canopy debirs, which constitute ideal substratum for colonization and survival of water-borne hyphomycetes. water from mist or rain flowing along the trunks (stemflow) and dripping from foliage (throughfall) are also major sources of conidia of water-borne hyphomycetes. about 33 species of fungi encompassing many water-borne hyphomycetes have been reported in rainwater dripping through building roofs in poland and predicted their survival due to accumulation of sediments on undulating roof surfaces (czeczuga, orłowska 1997). floral honey and honeydew excreted by the aphids in canopies are also serve as niches for fungi (magyar et al. 2005). so far, about 125 species of water-borne hyphomycetes have been reported from the tree canopies (sridhar 2009; karamchand, sridhar 2009; sridhar, karamchand 2009). the western ghats of india (8°20’–20°40’ n and 73°–77° e) as a hotspot of biodiversity endowed with moist deciduous and montane rain forests at an elevation up to 2000 m asl. the vegetation mainly consists of scrub jungle, grasslands, moist to dry deciduous forests, tropical evergreen forests and sholas. water-borne hyphomycetes have been explored from streams and rivers of coastal, mid-altitude (foothill) and mountain regions of the western ghats (e.g., sridhar, kaveriappa 1989; chandrashekar et al. 1990; sridhar et al. 1992; raviraja et al. 1998; rajashekhar, kaveriappa 2003). in view of limited studies on water-borne hyphomycetes of tree canopies in tropics (sridhar et al. 2006; karamchand, sridhar 2008; sridhar, karamchand 2009), the present investigation aims at exploring their assemblage and diversity in canopies of selected tree species during summer and monsoon seasons in kaiga forest stand located at the western ghats of southwest india. water-borne hyphomycetes 187 materials and methods the location selected for study is situated adjacent to the river kali near kaiga nuclear power station of the southwest india (~35 km east of the karwar city; ~55-70 m asl; 14°50′–14°51′ n, 74°26′ e). three trees each of five species (artocarpus heterophyllus lam., cassia fistula l., ficus recemosa linn., syzygium caryophyllatum (l.) alston and xylia xylocarpa roxb. taub.) distributed in an area of 300 m2 were chosen for study. the canopies of trees selected were free from interference of other trees. during april 30, 2009 (summer), partially decomposed trapped leaf litter at the tree holes or junctions of branches were sampled (the mean rainfall, temperature and humidity during april 2009 in kaiga forest was 0.7 mm, 30.8°c and 71.1% respectively). they were transported to the laboratory and rinsed in distilled water to remove adhered debris. small segments (5–8) weighing about 100–250 mg dry mass were aerated up to 48 h in 250 ml erlenmeyer flasks containing 150 ml of sterile distilled water (bubble chamber) at laboratory temperature (28±2°c). aerated water was filtered through millipore filters (5 μm; diam., 47 mm) to trap released conidia of water-borne hyphomycetes. the filters were stained with 0.1% aniline blue in lactophenol. each stained filter was cut in to half, mounted on a microscope slide with a few drops of lactic acid for screening conidia (nikon optiphot, japan: 20 ×, 40 ×, 100 ×) and they were identified based on conidial morphology using primary literature and monographs (ingold 1975a; nawawi 1985; marvanová 1997; gulis et al. 2005). sediments accumulated at the tree holes or junctions of branches sampled during april 30, 2009 were assessed for the presence of water-borne hyphomycetes by indirect method (sridhar et al. 2008). five pre-weighed sediment samples (200300 mg dry mass) were used (parallel samples were weighed before and after drying at 100°c for 24 h to convert wet to dry mass). each sediment sample was transferred to 250 ml erlenmeyer flask containing 150 ml sterile distilled water with 5 sterile banyan leaf (ficus benghalensis l.) disks (1.5 cm diam.). the flasks were incubated on a rotary shaker for 14 days (150 rpm, 28±2°c). the leaf disks were harvested, rinsed in distilled water to eliminate sediments and incubated in bubble chambers to stimulate production of conidia from colonized fungi and assessed further as described above. stemflow and throughfall of each tree were collected during july 30, 2009 (monsoon) (mean rainfall, temperature and humidity during july 2009 in kaiga forest was 1767 mm, 26.9°c and 98.2% respectively). shortly after the beginning of the rainy period (20–30 min), about 200 ml of water draining through the main stem was collected in sterile polythene bags and transferred to sterile glass bottles. a clean polythene sheet (2 m2) was spread below the canopy of each tree (~1 m above ground and ~3 m away from tree base), up to 200 ml of water dripping through each canopy was sampled and transferred to sterile glass bottles. within 30 min of sampling, aliquots (25 ml) of stemflow and throughfall were separately filtered through millipore filters to assess the free conidia of water-borne hyphomycetes. sediment moisture (gravimetric), ph and conductivity (1:2 dilution with distilled water) were assessed using water analysis kit (water analyzer 371, systronics, gujarat, india). similarly, temperature, ph and conductivity of stemflow and throughfall were assessed at the sampling site. parts of stemflow and throughfall samples 188 n. m. sudheep and k. r. sridhar were fixed at the site to estimate dissolved oxygen by winkler’s method (apha 1995). based on occurrence of water-borne hyphomycetes in trapped leaf litter, sediment, stemflow and throughfall, shannon diversity [h’ = – ∑ (pi × ln pi)] (magurran 1988) and pielou’s equitability (j′ = h′ ÷ h′max) (pielou 1975) were determined (where pi is the relative abundance of species i and h′max is the maximum value of diversity for the number of fungal species present). results average moisture content of the trapped leaf litter and sediments of the five tree species was 6.7% (range 5.5–8.1%) and 6.6% (range 5.1–8.1%) respectively (tab. 1). the mean temperature of stemflow and throughfall ranged between 24.4°c and 25°c respectively. the mean ph of sediment was acidic (6.1) than stemflow (7.1) as well as throughfall (7.6). sediments of xylea xylocarpa and syzygium caryophyllatum were acidic (4.8 and 5.1), while stemflow and throughfall of x. xylocarpa were alkaline (8.5 and 8.7). the mean conductivity of sediment was higher (288 μs/cm) than stemflow (60 μs/cm) and throughfall (61 μs/cm). average dissolved oxygen in stemflow and throughfall was 7.1 mg/l (range 6.9–7.5 mg/l). five canopy niches surveyed yielded a total of 29 species of water-borne hyphomycetes (tabs 2 and 3). ten species ranging from 2 (ficus recemosa) to 6 (cassia fistula) were recovered from the trapped leaf litter. six species ranging from 1 (xylea xylocarpa) to 3 (artocarpus heterophyllus and f. recemosa) were found in sediment. conidial output from trapped leaf litter and sediment was highest in syzygium caryophyllatum and least in x. xylocarpa (fig. 1). flagellospora curvula was found in table 1 physicochemical features of canopy samples of five tree species of kaiga forest assessed for water-borne hyphomycetes (n=3, mean) (ah, artocarpus heterophyllus; cf, cassia fistula; fr, ficus recemosa; sc, syzygium caryophyllatum; xx, xylia xylocarpa) parameter tree species mean (n=5)ah cf fr sc xx trapped leaf litter moisture (%) 7.5 5.6 8.1 6.9 5.5 6.7 trapped sediment moisture (%) 8.1 6.6 5.1 7.2 5.9 6.6 ph 6.5 7.1 7.1 5.1 4.8 6.1 conductivity (μs/cm) 302.3 75.3 276.7 554 232.7 288.2 stemflow temperature (°c) 24 24.5 25 25.5 26 25.0 ph 7.3 7.5 7.3 7.2 8.5 7.6 conductivity (μs/cm) 20 63.6 60.1 16.4 142 60.4 dissolved oxygen (mg/l) 7.5 7 7 7.2 7 7.1 throughfall temperature (°c) 24 24 25 25 24 24.4 ph 7.5 7.7 7 7.3 8.7 7.6 conductivity (μs/cm) 45.1 75.7 64.6 24.5 94.4 60.9 dissolved oxygen (mg/l) 7 7.4 6.9 7 7 7.1 water-borne hyphomycetes 189 table 2 percentage contribution of water-borne hyphomycete species to conidial production and diversity in trapped leaf litter and trapped sediment (in parenthesis) of five tree species of kaiga forest (ah, artocarpus heterophyllus; cf, cassia fistula; fr, ficus recemosa; sc, syzygium caryophyllatum; xx, xylia xylocarpa) taxon tree species ah cf fr sc xx flagellospora curvula ingold 13.9 6.1 25.8 1.4 9.2 triscelophorus acuminatus nawawi (71.4) (95.6) (96) (100) lunulospora curvula ingold 27.8 6.5 (28.6) 47 anguillospora longissima (sacc. & p. syd.) ingold 31.1 (8.1) 1.4 5.2 flagellospora penicillioides ingold 13.9 79 74.2 cylindrocarpon sp. 13.2 (65.7) (0.2) campylospora parvula kuzuha 53 29.4 tetracladium setigerum (grove) ingold 2.9 9.2 triscelophorus konajensis k.r. sridhar & kaver. (0.4) (4) campylospora filicladia nawawi 44.2 triscelophorus monosporus ingold (26.2) tripospermum myrti (lind) s. hughes 2.9 unidentified (tricladium-like conidia) 2.9 total species 5 (3) 6 (2) 2 (3) 4 (2) 5 (1) shannon diversity 2.215 (1.198) 1.216 (0.863) 0.824 (0.060) 1.179 (0.042) 1.886 pielou’s equitability 0.954 (0.756) 0.470 (0.863) 0.824 (0.038) 0.589 (0.242) 0.812 table 3 percentage contribution of water-borne hyphomycete species to conidial production and diversity in stemflow and throughfall (in parenthesis) of five tree species of kaiga forest (ah, artocarpus heterophyllus; cf, cassia fistula; fr, ficus recemosa; sc, syzygium caryophyllatum; xx, xylia xylocarpa) (*new record to canopy) taxon tree species ah cf fr sc xx anguillospora crassa ingold 34 (11.1) 50 (100) 16.7 (19.4) anguillospora longissima (sacc. & p. syd.) ingold (3.3) (28.6) 16.7 (40) 4.8 (2.2) triscelophorus acuminatus nawawi (53.3) 50 14.3 4.8 (7.5) lunulospora curuvla ingold (3.3) 33.3 (20) 76.2 (14.2) flagellospora curvula ingold 28.6 (20) 4.8 (14.9) ceratosporium cornutum matsush. 27.7 (14.3) (4.5) triscelophorus monosporus ingold (28.9) 16.7 4.8 helicomyces scandens morgan 6.4 16.7 trinacrium sp. (14.3) (2.2) flabellospora crassa alas. (13.3) (2.2) dwayaangam sp. 28.6 unidentified (sickle-shaped conidia) 28.6 dwayaangam cornuta descals (26.1) trisulcosporium acerinum h.j. huds. & b. sutton (20) flabellospora verticillata alas. (14.3) tricladium sp. (14.3) 190 n. m. sudheep and k. r. sridhar trapped leaf litter of all tree species, while triscelophorus acuminatus was more common in sediments of four tree species. fungal diversity in trapped leaf litter as well as sediment of a. heterophyllus was higher than other tree species. the species richness was higher in stemflow and throughfall than trapped leaf litter and sediments (23 vs. 13 species) (tab. 3). stemflow represented by 14 species ranging from 2 (cassia fistula) to 6 (artocarpus heterophyllus and xylea xylocarpa). throughfall consists of 16 species ranging from 1 (c. fistula) to 12 (x. xylocarpa). the conidial concentration in stemflow and throughfall was highest in x. xylocarpa and least in c. fistula (fig. 1). anguillospora crassa was common in stemflow of three out of five tree species, while anguillospora longissima was seen in throughfall of four tree species. fungal diversity was highest in throughfall of x. xylocarpa, while it was highest in trapped leaf litter in a. heterophyllus. discussion water-borne hyphomycetes in tree canopies have been explored extensively in temperate regions than tropical regions. niches in tree canopies investigated in temperate regions (mainly from canada, europe and japan) were gymnosperm needles, angiosperm leaves, tree hole, honey dew, melting snow, stemflow and throughfall (e.g., ando, tubaki 1984; czeczuga, orłowska 1998a, 1998b; magyar et al. 2005; gönczöl, révay 2003, 2004, 2006; sokolski et al. 2006). recently, a few niches in tree canopies (epiphytic fern, tree hole, stemflow and throughfall) of the west coast and western ghats of india have been studied (sridhar et al. 2006; sridhar, karamchand 2009; karamchand, sridhar 2009). the trapped leaf litter collected during summer consists of 10 species of waterborne hyphomycetes in our study. such trapped leaf litter in tree holes and epiphytic fern in sampaje of the western ghats during summer yielded 13 and 15 species (karamchand, sridhar 2008, 2009). however, conidial output from the trapped leaf litter in our study is considerably lower than sampaje region (35–192 vs. 363-601/g). as colonized fungi are viable in trapped leaf litter even during dry season, they will be disseminated within or across the canopy and also to aquatic bodies. helicomyces roesus link 12.8 *hydrometrospora symmetrica j. gönczöl & révay 12.8 flagellospora penicillioides ingold 6.4 diplocladiella scalaroides g. arnaud ex m.b. ellis 4.8 clavatospora tentacula sv. nilsson (2.2) alatospora acuminata ingold (2.2) triscelophorus konajensis k.r. sridhar & kaver. (2.2) total species 6 (5) 2 (1) 4 (6) 5 (4) 6 (12) shannon diversity 2.308 (1.678) 1.00 1.950 (2.518) 2.258 (1.922) 1.350 (2.985) pielou’s equitability 0.893 (0.723) 1.00 0.975 (0.974) 0.970 (0.961) 0.522 (0.833) tab. 3. cont water-borne hyphomycetes 191 fig. 1. conidial output of water-borne hyphomycetes from trapped leaf litter, trapped sediment and conidial load in stemflow and throughfall of five tree species of kaiga forest of the western ghats given in decreasing order (n=3, mean±sd) (tree species: ah, artocarpus heterophyllus; cf, cassia fistula; fr, ficus recemosa; sc, syzygium caryophyllatum; xx, xylia xylocarpa). 192 n. m. sudheep and k. r. sridhar the sediment analysis employed in our study is an indirect method to evaluate viable propagules (either spores or mycelia) of water-borne hyphomycetes capable of colonizing baited sterile leaf disks (sridhar et al. 2008). the species richness in sediments was lower than trapped leaf litter, stemflow and throughfall (6 vs. 10-16) reveals survival of a few species in tree sediments. triscelophorus acuminatus was most common in sediments collected from four tree species. similarly, t. acuminatus and t. konajensis were fairly dominant in sediments sampled from kaiga region (kaiga stream, kali river and kadra dam) (k.r. sridhar and n.m. sudheep; unpublished observations) indicates their survival under low o-r potential. these species were also dominant in trapped leaf litter in tree holes of sampaje region in spite of low dissolved oxygen in tree hole waters (1.14 mg/l) (karamchand, sridhar 2009). anguillospora longissma, flagellospora curvula, lunulospora curvula, t. acuminatus and t. konajensis were highly colonized the leaf disks immersed in tree holes of konaje region of the west coast of india in spite of low dissolved oxygen in tree hole waters (1.1–5.7 mg/l) (karamchand 2008). except for t. konajensis, rest of the species dominated in leaf litter or sediment, stemflow and throughfall in our study (see tabs 2 and 3). although the dissolved oxygen was low (3.8 mg/l) in a thermal spring of southern india, 14 species of water-borne hyphomycetes were recovered by chandrashekar et al. (1991). survival of aquatic hyphomycetes up to 12 months under anaerobic conditions has been demonstrated by field and webster (1983). depletion of oxygen from 94% to 4% saturation in microcosms resulted in 90% decrease in fungal biomass, the sporulation was strongly inhibited due to reduction of dissolved oxygen and the species richness was dropped to 60% (medeiros et al. 2009). viability of water-borne hyphomycetes in tree holes in spite of low dissolved oxygen during wet season and dry season facilitates recolonization of organic matter on the onset of wet conditions. the ph of sediment collected from xylia xylocarpa, syzygium caryophyllatum and artocarpus heterophyllus was acidic (4.8, 5.1 and 6.1). but the diversity of waterborne hyphomycetes was highest in a. heterophyllus and the leaf disks baited with sediment collected from s. caryophyllatum showed high conidial output. on the contrary, stemflow and throughfall of x. xylocarpa were alkaline (8.5, 8.7), but the species richness (6–12 species) and the shannon’s diversity (2.985) in throughfall were the highest among tree species studied. bärlocher (1987) compared canadian and european streams and demonstrated a slow or negligible decrease in species richness in circumneutral waters (5.7–7.2) with a rapid decline in alkaline waters (>7.2). raviraja et al. (1998) supported this view by demonstrating a negative correlation between species richness vs. ph in western ghat streams. the species richness in throughfall and stemflow was higher than trapped leaf litter and sediment (14–16 vs. 6-10 species) corroborates earlier study on water-borne hyphomycetes in nonriparian tree species of the west coast of india (sridhar, karamchand 2009). the ph of tree hole waters in sampaje region was close to neutral (6.98), while in our study, stemflow and throughfall were alkaline (7.2–8.5 and 7.3–8.7) (exception of throughfall of one tree species). the conidial concentration was highest in stemflow and throughfall of xylia xylocarpa in spite of alkaline ph (8.5 and 8.7). the conductivity was higher in sediment (288 μs/cm) as seen in tree hole waters of sampaje (148 μs/cm) (karamchand, sridhar 2008). compared to sediments and tree hole waters, the conductivity of stemflow and throughfall was low in our study water-borne hyphomycetes 193 (mean, 60–61 μs/cm) as evident in an earlier investigation (10.1–72.9 μs/cm) in the west coast (sridhar, karamchand 2009). three known and six unknown conidial morophotypes of dwayaangam species have been reported in rainwater samples (15 out of 25 tree species) in different locations of europe (gönczöl, révay 2006). interestingly, dwayaangam heterospora is known to parasitize eggs of rotifers and nematodes (barron 1991). dwayaangam colodena has also been reported as an endophyte in canopy needles of black spruce (picea mariana) (sokolski et al. 2006). in our study, dwayaangam cornuta and dwayaangam sp. contributed conidia between 26.1% (throughfall of xylia xylocarpa) and 28.6% (stemflow of ficus recemosa) respectively indicates their possible role as saprophytes, parasites and possibly endophytes in tree canopies. the multiradiate conidial shape of aquatic hyphomycetes is the product of convergent evolutions and secondary adaptation to aquatic mode of life (ingold 1975b; belliveau, bärlocher 2005). stone et al. (1996) suggested that canopy fungi may be the early colonizers of tissues in intact leaves and twigs, and act as a link between soil and aquatic food webs by completing their life cycles. studies on perfect-imperfect connections of aquatic hyphomycetes showed that the majority of species have evolved from ascomycetes found in decaying tree branches in streams (webster, descals 1979; webster 1992; marvanová 1997; sivichai et al. 2002; sivichai, jones 2003) and molecular studies also confirmed their terrestrial relatives (liew et al. 2002). selosse et al. (2008) proposed a hypothesis that a large numbers of conidia of water-borne hyphomycetes congregate in stream foam disperse through wind or aerosols to tree canopies facilitating their survival as epiphytes, endophytes or saprophytes. our study supports the view that at least a few water-borne hyphomycetes survive in trapped leaf litter and sediments in tree canopies in viable state for a considerable period during dry season facilitating their perpetuation on the onset of wet season. conidia in stemflow and throughfall during wet season are easily disseminated to other parts of the canopy or nearby aquatic habitats. widespread occurrence of water-borne hyphomycetes in tree canopies and their adaptation to wide range of canopy ecological conditions in temperate and tropical regions suggests their life cycle alternates between aquatic and canopy habitats. further studies are necessary to understand the functional significance of water-borne hyphomycetes, role in nutrient cycling in canopies and impacts of atmospheric pollution. acknowledgements. the authors are grateful to mangalore university for permission to carry out this study at the department of biosciences and nuclear power corporation of india ltd. 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(in:) b. kendrick (ed.). the whole fungus. national museums of canada, ottawa: 419–447. 2014-01-01t11:51:22+0100 polish botanical society syzygospora lapponica sp. nova (syzygosporaceae, heterobasidiomycetes) from finland heikki kotiranta1 and otto miettinen2 1finnish environment institute, research department p.o. box 140, fi 00251 helsinki, heikki.kotiranta@ymparisto.fi 2finnish museum of natural history, botanical museum, university of helsinki p.o. box 7, fi 00014, otto.miettinen@helsinki.fi k o t i r a n t a h., m i e t t i n e n o.: syzygospora lapponica sp. nova (syzygosporaceae, heterobasidiomycetes) from finland. acta mycol. 41 (1): 21 24, 2006. a new syzygospora species from finland, s. lapponica is described and illustrated. the hitherto collections derive from finnish lapland and the species is apparently a mycoparasite of the rare old growth forest dwelling polypore antrodia infirma. the new species deviates from other species in the genus in having cylindrical, slightly bent spores and having a polypore as the host. key words: antrodia infirma, lapland, old-growth forest, syzygospora introduction according to the index fungorum (2006) the genus syzygospora g. w. martin contains 15 species of which four have previously been reported from finland: s. bachmannii diederich & m. s. christ., s. mycophaga (m. p. christ.) hauerslev, s. pallida (hauerslev) ginns and s. tumefaciens (ginns & sunhede) ginns (k o t i r a n t a , l a r s s o n 1990; k o t i r a n t a , s a a r e n o k s a 1993, 2000; k o t i r a n t a 2001; h a r m a j a 2003). according to e.g., g i n n s (1986), r o b e r t s and h a u e r s l e v (1997) or c h e n et al. (1998) none of the syzygospora species is known to be a mycoparasite of polypores. also the spores of most of the species are basically ellipsoid, thus differing from those seen in the new species. materials and methods thirty spores per specimen are measured, and the measurements are made in cotton blue (cb) or melzer´s reagent (iki). cb– means that the walls of the cells are not stained by cotton blue, and cb+ that they are stained, and iki– that there is no reaction to melzer´s reagent. acta mycologica vol. 41 (1): 21-24 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 22 h. kotiranta and o. miettinen the following abbreviations are used: l* mean spore length, w* mean spore width, q range of the variation in l/w ratio, q* quotient of the mean spore length and width (l/w). none of the measurements derive from spore print. biological provinces and collecting sites in finland are indicated according to the finnish national uniform grid system (27°e), as applied to biological material by h e i k i n h e i m o and r a a t i k a i n e n (1981). syzygospora lapponica miettinen & kotir., sp. nova holotype: finland. sompion lappi: sodankylä, raitiojänkä, mikkelinpuro, kivivärriön kummut, pine dominated old-growth forest on poor soil, inside antrodia infirma renvall & niemelä on decorticated, fairly advanced decayed, 25 cm thick pinus sylvestris, 67°38´n, 27°20´e, (grid 27°e:7505809:514633), 3 oct 2005 miettinen 10748 (h). fructificatio invisibilis; systema hypharum monomiticum; hyphae fibulatae; cystidia desunt; basidia cylindracea vel sinuosa, tetrasterigmatica; conidia ellipsoidea vel cylindracea, 5–7 x 2.5–3 μm; sporae cylindraceae, 5–6 x 2 μm. fig. 1. syzygospora lapponica miettinen & kotir. (a b drawn from miettinen 10748, type, c from miettinen 10780): a section trough basidiocarp showing a hyphal peg, conidiophores, conidia, basidia and basidiospores; b spores; c spores. syzygospora lapponica sp. nova (syzygosporaceae, heterobasidiomycetes) 23 basidiocarp invisible. hyphal system monomitic, hyphae clamped, in subiculum 2–3 μm wide, thin-walled, in subhymenium 3–4 μm wide, very thin-walled, cb–, iki– . cystidia none, but sterile, apically slightly widened hyphal ends form hyphal pegs which penetrate to the tubes of the host. no haustoria observed. conidiophores abundant, clamped, 2–3 μm in diam., very thin-walled. conidia born in the apices of conidiophores, ellipsoid or cylindrical, (4.2-)5-7(-7.3) x (2.3-)2.5-3(-3.7) μm, relatively thin-walled, cb–, iki–. basidia solitary between the conidiophores or forming a more or less continuous hymenium, cylindrical or sinuous, basally clamped, very thin-walled, (16-)20-30(-32) x 4-5 μm, with four, up to 4 μm long, very thin, needle-like sterigmata. spores cylindrical, sometimes slightly bent, 5-6.3(-7) x (1.6-)1.8-2.1 μm, l* 5.7 μm, w* 1.9 μm, q 2.5-3.7, q* 3, (miettinen 10780), 4.5-5.6(-6.2) x 1.7-2 μm, l* 5.1 μm, w* 1.9 μm, q 2.4-3.3, q* 2.7 (miettinen 10748, type), with a negligible apiculus, very thin-walled, cb-, iki-. additional specimen examined: finland. sompion lappi: savukoski, mukkajoenrovat, välirova, dry pine dominated old-growth forest, inside antrodia infirma on long time ago burned, heavily charred, decorticated, fairly advanced decayed, 32 cm thick pinus sylvestris, 67°56´n, 28°20´e (grid 27°e:7540124:555948), 4 oct 2005 miettinen 10780 (h). material for this study was collected during inventories of unprotected, stateowned old-growth forests in finnish lapland. the two hitherto finds derive from pine dominated old-growth forests on poor soils, where they grew inside basidiocarps of the polypore antrodia infirma. these forests with abundance of kelo pine trees (n i e m e l ä et al. 2002), dry microclimate and history of forest fires form a special kind of ecosystem that used to typify northeastern fennoscandian forest landscapes. they harbour a number of specialist species adapted to the harsh ecological conditions such as a. infirma and ecologically closely related a. primaeva renvall & niemelä. tens of basidiocarps of both a. infirma and a. primaeva have been studied during these inventories, as well as specimens of a. crassa (p. karst.) ryvarden, a. serialis (fr.) donk, a. sinuosa (fr.) p. karst. and a. xantha (fr.: fr.) ryvarden from the same area. with only two finds, it seems that syzygospora lapponica is a rare species, and possibly restricted to a. infirma and the dry old-growth pine forests of the north. the host a. infirma itself is considered rare and classified as a vulnerable (vu) species in finland (r a s s i et al. 2001). if a. infirma is the sole host species of s. lapponica, also it should be classified as a threatened species. acknowledgements. we are grateful to teuvo ahti (helsinki) who helped us with the latin diagnosis. a research grant from the ministry of environment (ym131/5512/2002) helped us to carry out this study. references c h e n c j., o b e r w i n k l e r f., c h e n z c. 1998. syzygospora nivalis sp. nov. from taiwan. mycotaxon 67: 217 226. g i n n s j. 1986. the genus syzygospora (heterobasidiomycetes: syzygosporaceae). mycologia 78: 619 636. h a r m a j a h. 2003. syzygospora tumefaciens (fungi: tremellales) found in finland. memoranda soc. fauna flora fennica 79: 73 74. 24 h. kotiranta and o. miettinen h e i k i n h e i m o o., r a a t i k a i n e n m. 1981. ruutukoordinaattien ja paikannimien käyttö suomessa [grid references and names of localities in the recording of biological finds in finland]. notul. en tomol. 61: 133 154. [in finnish]. index fungorum 2006. syzygosporaceae. www.indexfungorum.org/names/names.asp, 1 feb. 2006. k o t i r a n t a h. 2001. the corticiaceae of finland. publ. botany univ. helsinki 32: 1 29. k o t i r a n t a h., l a r s s o n k h. 1990. new or little collected corticolous fungi from finland (aphyl lophorales, basidiomycetes). windahlia 18: 1 14. k o t i r a n t a h., s a a r e n o k s a r. 1993. rare finnish aphyllophorales (basidiomycetes) plus two new combinations in efibula. ann. bot. fennici 30: 211 249. k o t i r a n t a h., s a a r e n o k s a r. 2000. corticioid fungi (aphyllophorales, basidiomycetes) in finland. acta bot. fennici 168: 1 55. n i e m e l ä t., wa l l e n i u s t., k o t i r a n t a h. 2002. the kelo tree, a vanishing substrate of specified wood inhabiting fungi. polish bot. jour. 47: 91 101. r a s s i p., a l a n e n a., k a n e r v a t., m a n n e r k o s k i i. (eds) 2001. suomen lajien uhanalaisuus 2000 [the 2000 red list of finnish species]. ministry of environment and finnish environment institute, helsinki. [in finnish]. r o b e r t s p., h a u e r s l e v k. 1997. syzygosporaceae jülich. (in:) h a n s e n, l., k n u d s e n, h. (eds) nordic macromycetes 3. heterobasidioid, aphyllophoroid and gastromycetoid basidiomycetes. nordsvamp, copenhagen: 84 86. syzygospora lapponica sp. nova (syzygosporaceae, heterobasidiomycetes) z finlandii s t r e s z c z e n i e nowy gatunek syzygospora lapponica zosta opisany i zilustrowany. kolekcja pochodzi z fińskiej laponii, gdzie grzyb ten występuje jako pasożyt na owocnikach antrodia infiryna w starych lasach. opisany gatunek różni się od innych z tego rodzaju zarodnikami w kształcie cylindrycznym, lekko wygiętymi oraz występowaniem na poliproidalnym gospodarzu. 2014-01-01t11:43:12+0100 polish botanical society global warming and mycoflora in the baltic region hanns kreisel zur schwedenschanze 4, d 17498 potthagen, hanns.kreisel@gmx.de k r e i s e l h.: global warming and mycoflora in the baltic region. acta mycol. 41 (1): 79 94, 2006. the author discusses possible effects of global warming on distribution and ecology of larger fungi, and presents examples of suggested indicator species which apparently are spreading from south to north. only basidiomycetes are corncerned, while actually no case of non lichenized ascomycetes is known. a continued monitoring of the mentioned species is recommended. key words: mycoflora, basidiomycetes, global warming, baltic region introduction global warming (climatic change) with its consequences for weather and local climate, rising sea level, retreat of glaciers and of polar ice calottes, and subsequent nature catastrophes, actually is much disputed in newspapers, journals, and book publications. highly reputed specialists of climatology, oceanography, or physics, have investigated the phenomena (e. g. r a h m s t o r f , s c h e l l n h u b e r 2006). since the end of last ice age, some 15 000 years ago, climate in central and northern europe is warming, and enormous ice calottes have retired from this region, making possible a re-settelement of large areas by vegetation and fauna. this process was not continuous, but interrupted by phases of standstill alternating with phases of further warming. nevertheless, the actual phase of relatively fast warming is regarded as man-made to a large extent, and therefore requires special attention by scientists. also in mycology, we are contemporary witnesses of changes in distribution and ecology of fungi, and we should use the chance to observe and describe the corresponding evolutions. during the last decenniums, attention of mycologists was concentrated on processes of threat and decline of mycoflora by presumably man-made factors, with subsequent publication of a rich literature of red data lists and evaluations of long registers of species regarded as endangered in different degrees. acta mycologica vol. 41 (1): 79-94 2006 dedicated to prof dr. alina skirgiełło, warszawa, with motivation of her 95th birthday 80 h. kreisel this is not the place to discuss whether or not it is possible to conserve the mycoflora of a given territory in its actual composition and status, but we should be aware that we are living on a planet which is in permanent evolution since thousands millions of years, and will continue to evolve. up to date, only a few mycologists, and only in a few special cases, have discussed the consequences of global warming. but there is an interesting paper of dutch lichenologists (h e r k et al. 2002) who have found, basing on a long-term monitoring, that many tropical and subtropical species of epiphytic and terrestrial lichen species are invading the netherlands, while already 50% of the arctic-alpine and boreomontane species shows a decline. if we are looking for comparable processes in non-lichenized larger fungi, we only can form a general picture by comparison of many small detailed informations. judging from experiences and publications from central and northern europe, in particular from countries around the baltic sea, the following impressions raise: 1. a number of species is moving from southern countries (mainly the submediterranean region) to north. but this is not an advance on broad front, but only in more or less isolated localities, volatile, and interrupted by temporary retreats. 2. main fructification time of macromycetes is shifting from late summer and early fall to late fall, even beginning winter. this remembers the situation in regions with mediterranean clima. organizers of forays, mushroom exhibitions and similar events already had to note this phenomenon by success or failure of their efforts. 3. consequences of climate on annual mushroom yield have been supposed by r i c h t e r (2005, 2006). in an area north of berlin he demonstrated a positive trend in the period 1977 2003 for cantharellus cibarius, boletus edulis, sparassis crispa, and gyroporus cyanescens, while the yield of xerocomus badius and amanita rubescens was showing a decrease. on increasing frequency of the poisonous agaricus xanthodermus see above. in the following lines, only the first aspect shall be considered and substantiated by some facts and data. wood inhabiting macromycetes auricularia mesenterica (dicks. : fr.) pers. was known in central germany already in the 19th century, but only since 1970 ist is known in northern germany (baltic coast), and 1983 in lower elbe valley (w ö l d e c k e 1998; o t t o et al. 2005 map). the actual extension of its area has reached southern norway and central sweden (r y m a n , h o l m å s e n 1984, map). coriolopsis trogii (berk.) dom. [trametes trogii berk.] this species seems to be indigenous in the rhine valley from sw. germany to netherlands, in the inn valley of se. bavaria, in moravia and slovakia (k o t l a b a 1984, map; k r i e g l s t e i n e r 1991, map; a r n o l d s et al. 1995, map). outposts in central and northern germany were noted 1945 near alsfeld, hessen and 1961 near detmold (k r e i s e l 1962), 1977 in berlin (k a s p a r 1979), 1996 near merseburg (r i c h t e r , s c h u r i g 1997), 1998 near borna, saxonia (h a r d t k e , o t t o 1998), and 2004 near braunschweig (a n d e r s s o n 2004). in poland the species has reached central poland (n i e m e l ä et al. 1992), and in scandinavia isolated localities in southern norway, central sweden and south-eastern finland (o l o f s s o n global warming and mycoflora in the baltic region 81 1996, map; ve s t e r h o l t in h a n s e n , k n u d s e n 1997; l a r s s o n 1997, map). up to date, there seems to be no record from denmark. diplomitoporus flavescens (bres.) dom. [trametes flavescens bres.]. this species, bound to pinus spp., was known around 1970 to occur north to southern germany, eastern saxonia (d u n g e r 1987), southern and eastern poland (wo j e w o d a 2003); it was discovered 1974 in southern brandenburg (eastern germany), 1983 in berlin and after 1994 north of berlin (b e n k e r t 2004, maps), it was found before 1983 in the słowinski national park, northern poland (b u j a k i e w i c z , l i s i e w s k a 1983), and 1986 in the netherlands (arnolds & al. 1995). in sweden, only 3 localities have been noted before 1996 (o l o f s s o n 1996, map). ganoderma adspersum (schulzer) donk [g. australe (fr.) pat. agg.] is considered as an expanding species by several authors (arnolds & al. 1995 for netherlands, k r e i s e l 1998, with map, for eastern germany, wo j e w o d a 2003 for poland). although known from prague as early as 1851 (k o t l a b a 1984, map), it was found only 1959 near detmold, 1960 near potsdam, 1961 near dresden, 1964 near stralsund, 1981 on fyn and sjaelland (denmark), 1986 on the frisian island langeoog, 1996 at the southern coast of sweden, and 1997 in oslo, norway (j a h n 1963; p e t e r s e n 1983; k r e i s e l 1987; o l o f s s o n 1996, map; k r e i s e l 1998, map; w ö l d e c k e 1998). ganoderma pfeifferi bres. the species occurs mainly on liv-ing truncs of fagus sylvativa. it was known already 1860 near greifswald and 1864 on lolland, denmark (p e t e r s e n 1983; k r e i s e l 1998, map), but was recorded only 1971 ff in skåne, southern-most sweden (b o h l i n 1971), and later in blekinge and väster-götland, sweden (o l o f s s o n 1996, map; l a r s s o n 1997). in poland its distribution probably is synanthropic (wo j e w o d a 2003). ganoderma resinaceum boud. this another southern ganoderma species “going north”. it was recorded 1910 in southern moravia (k o t l a b a 1984), 1930 in eastern saxonia, 1932 near darmstadt, hessen, 1943 ff in the netherlands (after 1970 increasing there), 1950 in lolland, denmark, 1963 westfalen, 1981 ff in niedersachsen, germany, 1987 in halland, sweden, but only 1995 ff in rostock and other places along the baltic coast in north germany (j a h n 1963; p e t e r s e n 1983; a r n o l d s 1995; o l o f s s o n 1996, map; k r e i s e l 1998, map). hirneola auricula-judae (bull. : fr.) wettst. [auricularia auricula-judae (bull. : fr.) berk.] is actually quite common in germany and poland and was recorded as early as 1721 in central germany and 1753 in eastern brandenburg (t ä g l i c h 1998; b e n k e r t 2004), but there is a distinct increase of its frequency after 1970 in northern germany (b e n k e r t 2004 with maps; kreisel n. p.). a significant spreading was noted after 1945 in denmark and southern sweden (knudsen & pedersen 1980, maps) and after 1980 in the netherlands (n a u t a , ve l l i n g a 1995). inonotus nidus-pici pilát was observed since 1887 in czechoslovakia (k o t l a b a 1984), but appeared 1967 in potsdam, 1977 near eberswalde, 1978 near parchim, 1984 ff in and around demmin, and 1985 near rostock, all in northern germany (b e n k e r t 1971; k r e i s e l 1994, map). strangely enough, there is no record of this submediterranean species from the neighbour regions poland and western germany, and there are no more records from germany after 2000. up to date the species was not recorded from denmark and fennoscandia. 82 h. kreisel mutinus caninus (huds. : pers.) fr. the species is common in forests of central europe and denmark for a long time, but its immigration in scandinavia is described by a n d e r s s o n (1941). actually it is rare in norway and sweden, and known from only one locality in south-western finland (e c k b l a d in h a n s e n , k n u d s e n 1997). pluteus aurantiorugosus (trog) sacc. [p. coccineus (massee) j. e. lange] is rare, but widely distributed in central europe, in the netherlands and in southern poland (k r e i s e l 1987; k r i e g l s t e i n e r 1991, map; a r n o l d s 1995; wo j e w o d a 2003), being more frequent only in south-western germany and the rhine valley. it was recorded in jylland, denmark, 1917 ff (l a n g e 1936), but only since 1984 in northern germany (k r e i s e l 1987; o t t o et al. 2005, map). in recent times it appeared in sweden from skåne to västergötland and öland (l a r s s o n 1997), and 2002 in norway near oslo (g u l d e n 2002). polyporus alveolaris (dc. : fr.) bondartsev & singer [p. mori pollini: fr., favolus europaeus fr.] has been found already in the 19th century in czechoslovakia (pilát 1936; k o t l a b a 1984). in germany it was recorded 1929 in the black forest (schwarzwald) and is now widely distributed in south-western germany and eastern bavaria, extending north to the main valley and the eifel mts. (k r i e g l s t e i n e r 1991, map; l o h m e y e r 2003). in poland it was found 1999 near tarnów and recently along odra river near kostrzyn and szczecin (wo j e w o d a 2003). up to date, there are no records from north germany, denmark, and scandinavia. sarcodontia crocea (schwein. : fr.) kotl. [s. crocea (pers.) donk] has penetrated in two periods to the baltic region. in the 19th century, the northern limit of its distribution was marked by göttingen, dessau, görlitz, legnica, wroclaw, kraków, and bieszczady mts. (k r e i s e l 1991; wo j e w o d a 1973, 2003), but around 1900 several records were noted around berlin, 1902 on öland (sweden), moreover near gamborg (fyn, denmark), 1928 near lublin in poland and 1933 near elbing (elbląg). all of these outposts were extinct for several decenniums, but in 1970 another advance started in ne germany: 1970 near guben, 1970 and 1979 rostock, 1975 potsdam, 1980 neubrandenburg, 1983 berlin-mahlsdorf, 1990 greifswald and near wismar. simultaneously, the species appeared in the netherlands (prov. zeeland) in 1975 and 1984 (a r n o l d s et al. 1995). in southernmost finland now the 61° latitude is reached (s c h u m a c h e r 2005). schizophyllum commune fr. : fr. although the common split-gill was observed in central europe and in denmark already in the 18th century, there was a notable increase in frequency and distribution since 1932 in denmark and since 1948 on bornholm island (b j ø r n e k a e r , b u c h w a l d 1933; k n u d s e n , p e d e r s e n 1983, maps), after 1950 in northeastern germany (d ö r f e l t et al. 1988, map). the actual distribution reaches southern norway, southern and central sweden and southern finland (r y m a n , h o l m å s e n 1984, map; k ä ä r i k in h a n s e n , k n u d s e n 1997). in contrary, a decrease was noted since 1980 in the netherlands (n a u t a , ve l l i n g a 1995). less distinctive appear the cases of some wood-inhabiting saprobic agarics and boletes, regarded as “southern species”, as haasiella venustissima (fr.) kotl. & pouzar, pulveroboletus hemichrysus (berk. & m. a. curtis) singer, rhodotus palmatus (bull. : fr.) maire. these deserve further attention and monitoring, for they have been noted occasionally and inconsistently in northern germany and scandinavia. global warming and mycoflora in the baltic region 83 h. venus-tissima and rh. palmatus have been observed in sweden already in the 19th century. such early outpost records may be interpreted as relicts of a former warming period but in that time, number of observers was too low, and mapping was not applied, therefore no clear evidence is available. another wood-inhabiting species to be monitored carefully is plicatura crispa (pers. : fr.) rea [p. faginea p. karst.] for actually it is advancing (or re-settling lost areas ?) in central and northern germany, but it is not clear whether or not this can be interpreted as consequence of global warming. soil and litter inhabiting saprobes agaricus bohusii bon, a species occuring mainly in alluvial forests and on synanthropic habitats, was observed since 1922 in bohemia, but only since 1969 in hungary, 1978 in central germany near halle , 1981 near magdeburg, before 1984 in poland near bielsko-biała, 1986 in hannover, 1990 near leipzig, 1993 near zittau, 2000 near wismar (h e r r m a n n 1987; w ö l d e c k e 1998; wo j e w o d a 2003; s p e c h t 2005; o t t o et al. 2005, map); in denmark it was recorded since 1993 in copenhagen, and since 1998 near aarhus, jylland (p e t e r s e n 2006). it was rarely found in the netherlands (a r n o l d s et al. 1995), but still not recorded from scandinavia. agaricus xanthodermus genev., the poisonous yellow-staining mushroom, is common in nearly the whole germany (k r i e g l s t e i n e r 1991, map; o t t o & al. 1994, map), netherlands, denmark, poland (s k i r g i e ł ł o 1986, map), hungary etc., but rare in southern scandinavia (h a n s e n , k n u d s e n 1992; r y m a n , h o l m å s e n 1984, map). recently, several observers in central and northern germany have noted a discinct increase in frequency of this species. clathrus archeri (berk.) dring [anthurus archeri (berk.) e. fisch.] is a frequently documented and mapped case. the species, apparently introduced from australia, appeared 1914 in eastern france (vosges mts.), 1938 in germany near karlsruhe (rhine valley), 1942 near basel in switzerland, 1948 in austria, 1965 in western bohemia, 1985 in southern poland and 1989 in slovakia. in germany, the species extended continuously to north: 1953 it was found in the main valley, an important biogeographic line (s t r i c k e r 1954, maps), 1960 near querfurt in central germany (h e r r m a n n 1962, 1971), 1963 near meißen (saxonia), 1977 in the harz mts. and in rostock near baltic sea, 1981 ff lower saxonia, 1984 ff near greifswald. there is one record from denmark (fyn), but only an erroneous record 1942 in norway. in the netherlands the species appears since 1973 and is spreading (arnolds & al. 1995). the early appearence of the species since 1945 in cornwall, southwestern england, may be due to another introduction. for a rather actual map of distribution in europe see k r e i s e l (2004), for poland s t e n g l -r e j t h a r , wo j e w o d a (1985), eastern germany d ö r f e l t et al. (1988), czechoslovakia k l u z á k (1990), western germany k r i e g l s t e i n e r (1991). c. archeri meanwhile is naturalized in forests in wide parts of central europe, whereas its habitats in northern germany are mainly cemeteries, parks, recreational forest areas near towns, and other synanthropic places. conocybe intrusa (peck) singer [c. hebelomatoides middelhoek & reijnders]. the strange agaric, possibly introduced from north america, was observed in eu84 h. kreisel rope first 1950 in hengelo, netherlands (a r n o l d s et al. 1995), 1968 and 1971 in berlin (b e n k e r t 2005) and then in several places around berlin and in central germany, including 1977 zittau (h e r r m a n n 1976; e d e r 1983; d ö r f e l t et al. 1988, map), 1979 ff an a few places in lower saxonia (w ö l d e c k e 1998), 1987 near greifswald, and 1997 wyszków in central poland (wo j e w o d a 2003), but already 1973 in lithuania (u r b o n a s et al. 1986). many of the records are from glasshouses, but others from gardens, vegetable waste, manure heaps, and other open places. lycoperdon marginatum vittad., a species of open grassland on gravelly or sandy soil, of mainly southern distribution, was rare everywhere in central europe. in northern germany it was found 1902 and 1905 around berlin and rediscovered 1970 near potsdam, 1984 near görlitz, and 2003 near zessin on the rügen island (b e n k e r t 1973; h a r d t k e , o t t o 1998; k r e i s e l 2005). mutinus elegans (mont.) e. fisch. [m. curtisii (berk.) e. fisch., m. inopinatus ulbrich] is another species of north american origin. in europe it was first noted 1929 at lago maggiore in northern italy (s t o m p s 1931), 1936 in lower rhine valley in north-western germany (u l b r i c h 1937), 1948 near karlsruhe, 1977 ff in saxonia, 1982 ff in berlin, 1984 ff near pasewalk in north-eastern germany, the last being actually the northernmost locality in europe (k r e i s e l 1987; r a u s c h e r t , h e l l m u n d 1989; h a r d t k e , o t t o 1998; k r e i s e l 2004, map). m. elegans was observed in germany mainly in gardens, frequently associated with tall grasses (miscanthus spp., arundo donax, b e n k e r t 2005). in the netherlands it was found only 1989 near ede (a r n o l d s et al. 1995; n a u t a , ve l l i n g a 1995). there are no records from denmark, scandinavia, and poland. mutinus ravenelii (berk. & m. a. curtis) e. fisch. was equally introduced from north america and was noted at first 1942 in berlin. in contrary to m. elegans, it expanded mainly to north and was found after 1950 in the netherlands, 1960 near hamburg, 1961 in riga and southern finland (here 1981 already in oulu), 1968 1984 again in berlin (b e n k e r t , j e n t s c h 1985), and is knowm from rare collections in denmark, sweden, and norway. in germany, a certain concentration of localities has established around hamburg, and in the spreewald estuary se of berlin. in eastern europe, m. ravenelii is known from bohemia (k u t h a n , ve s e l s k ý 1967; s e d l á č e k , s k á l a 1985; b í c h a 1986), 1969 ff from southern and central poland (g u m i n s k a 1985; s o k ó ł , s z c z e p k a 1987, map), about 1980 from latvia and estonia (v i m b a , ya r v a 1981; j ä r v a , v i m b a 1981). a map of distribution in europe was published by k r e i s e l (2004). in summary, most saprobic soil-inhabiting species recorded in part b are introduced from other continents or of unknown origin. ectomycorrhizal fungi amanita phalloides (fr.) link is a frequent species in most of central europe, distributed since old times from submedi-terranean region till baltic sea, e. g. in germany, poland, denmark, and the baltic states. in fennoscandia, it is restricted to the fagus and quercus belt, north till hordaland in norway, southern sweden, åland islands, and south-western finland. the first record near oslo was registered only 2000 by b r a n d r u d (2001), what suggests a slight extension to north in norway. in contrary, in nort-western germany it is scarce (k r i e g l s t e i n e r 1991, map), global warming and mycoflora in the baltic region 85 and in the netherlands there is a significant decline (n a u t a , ve l l i n g a 1995, but see a r n o l d s et al. 1995). amanita solitaria (bull. : fr.) mérat ist frequent in the warmer parts of southern germany, but rare in central germany north to the harz region, and very rare in the netherlands. in northern germany it was discovered 1989 near braunschweig (w ö l d e c k e 1998), 1997 near templin (d o l l 2001), and 2002 near schwerin (k r e i s e l 2004). distribution in western germany was mapped by k r i e g l s t e i n e r (1991). the species is not known from poland and fennoscandia. amanita verna (bull. : fr.) roques should not be confused with albinotic forms of a. phalloides. the true a. verna is a more thermophilous mushroom which occurs in germany only north to saarland and the main valley (k r i e g l s t e i n e r 1991, map). it does not occur in the netherlands (a r n o l d s et al. 1995) and probably not in poland (wo j e w o d a 2003), but it was recently introduced to saxonia and caused there (near bautzen) in 2005 a severe case of poisoning, recorded in television and newspapers; the fungi were determined by the mycologist g. zschieschang (w. tietze in litt.). the case shows how advance of southern species has implications even to toxicology and popular mycology. amanita vittadinii (moretti) vittad. is distributed in europe mainly in the mediterranean region. since 1942 it was observed in netherlands near delft, where it is stable for more than 50 years within a very limited area (a r n o l d s et al. 1995). since 1986 is was observed in central germany near merseburg (d ö r f e l t 1989), and 1992 it was found in hannover (w ö l d e c k e 1998). up to date, there is no record from northern germany, poland, and fennoscandia. chalciporus rubinus (w. g. sm.) singer [boletus rubinus w. g. sm.]. although originally described in 1868 from england, the main distribution of this rare species seems to be in france, italy, austria, hungary, southern moravia, and bohemia (p i l á t , d e r m e k 1974). since 1933 it was observed repeatedly in saxonia in and near dresden, 2000 ff. in leipzig (k l e i n e et al. 2005), 2005 in rühstädt at lower elbe (d. benkert in litt.); in the netherlands it was found 1993 in utrecht (a r n o l d s et al. 1995). all records from germany and the netherlands are from old parks on alluvial riverside habitats. cortinarius orellanus fr. [dermocybe orellana (fr.) ricken]. this is another poisonous mushroom which actually seems to be in expansion. in germany it is rather frequent in the south-west, more rare in the other parts, and rare in the north (k r i e g l s t e i n e r 1991, map; o t t o et al. 1994, map). in poland it is scattered mainly in southern and eastern poland, but was recorded recently near szczecin (wo j e w o d a 2003). it is rare in denmark and southern sweden (north to bohuslän and gotland, l a r s s o n 1997), but a recent advance in southern norway was documented by b r a n d r u d (2001, map). pulveroboletus gentilis (quél.) singer [p. cramesinus (secr. ex gilbert) m. m. moser] is regarded as thermophilic and calciphilic in germany, where it occurs scattered in the southern and central parts (k r i e g l s t e i n e r 1991, map; o t t o et al. 1994, map), whereas it is rare in poland (wo j e w o d a 2003) and in the netherlands (n a u t a , ve l l i n g a 1995 “regarded as threatened”, map). in northern germany recently a few localities have been recorded near lübeck (u n g e r 1998) and 2003 near schwerin (b. westphal in litt.). isolated localities are known from denmark and norway (k n u d s e n in h a n s e n , k n u d s e n 1992), whereas in sweden it is 86 h. kreisel known from several places from skåne north to götaland and stockholm (l a r s s o n 1997, map). it is not clear, whether or not a recent expansion is occurring in northern germany and scandinavia. scleroderma cepa pers. the thermophilic species is rare in whole germany, poland, and netherlands, but is known also from denmark and southern sweden (h a n s e n , k n u d s e n 1997). some more recent records from lower saxonia 1969 ff (w ö l d e c k e 1998), near lübeck 1982 (u n g e r 1994) and 2005 in south-western saxonia (ch. morgner in litt.) suggest a slight increase in frequency in central and northern germany, but in northern poland it was recorded by te o d o r o w i c z already 1939 near gdańsk. few mycologists in central europe are really familiar with this species, but the useful and illustrated keys published recently in italy and spain will facilitate a more thorough study of the distribu-tion of s. cepa in next future. fungi of xerothermic grassland endoptychum agaricoides czern. the secotioid mushroom, widespread in xeric climates of mediterranean region, se. europe and central asia, goes north to aoutheastern moravia and southern slovakia. only two outposts in the area considered here have been stated: 1941 near sollentuna, uppland in central sweden (a r w i d s s o n 1945; l a r s s o n 1997), and 1985 near potsdam in eastern germany (k r e i s e l 1995). both localities were ephemerous and apparently independent of each other. floccularia straminea (p. kumm.) pouzar [armillaria luteo-virens (alb. & schwein. : fr.) gill. ss. auct. mult.]. this agaric occurs scattered in bavaria and in central germany till north of harz mts. (h e r r m a n n 1971; k r i e g l s t e i n e r 1991, map; g r ö g e r in k r e i s e l 1987), in poland only in tatry national park (wo j e w o d a 2003), but is lacking in the netherlands, whole northern germany, and most of poland, but a very few temporary outposts have been observed in scandinavia: one in buskerud, southern norway (g u l d e n in h a n s e n , k n u d s e n 1992) and two in sweden: about 1940 near stockholm, and 1988 at visby on gotland (b o h u s j e n s e n et al. 1990; l a r s s o n 1997). gastrosporium simplex mattir. the “steppe truffle” was observed since 1956 in many localities in central germany, from thuringian bassin till north of harz (r a u s c h e r t 1962, map) and in the rhine-main disctrict (k r i e g l s t e i n e r 1991, map). very few records exist from north-eastern germany, where it was found at seelow near oder and at altentreptow (d ö r f e l t , r a u s c h e r t in k r e i s e l 1987). in poland, there are records from southern poland and again in the odra region at bielinek nad odrą (š m a r d a 1957) and pyrzyce (wo j e w o d a 2003). since 1980 the species is known from a few localities in sweden: skåne and västergötland (k e r s 1980; l a r s s o n 1997). geastrum pseudolimbatum hollós. our knowledge about this species is incomplete, for it was not distinguished from g. coronatum pers. by several authors. it is very rare in xero-thermic regions of eastern germany (d ö r f e l t et al. 1979, map), but lacking in western germany. there are two records in netherlands (1929 and 1980, a r n o l d s et al. 1995) and two in sweden (skåne and västergötland, s u n h e d e 1989; l a r s s o n 1997), but none from poland. montagnea radiosa (pallas) šebek [m. arenaria (dc.) zeller]. a frequent inhabitant of continental steppes and semideserts, it was discovered 1964 near mücheln in global warming and mycoflora in the baltic region 87 central germany, where it still was confirmed in 1995 and 1996, and in a few other localities near erfurt and halle (r a u s c h e r t 1964; d ö r f e l t 1974, map; k r e i s e l 1987; d ö r f e l t , r i c h t e r 1996). further outposts exist in poland near bielinek nad odrą (s k i r g i e ł ł o 1977) and more recently near pyrzyce in northwestern poland and near przemysl (wo j e w o d a 2003). up to date, the species is not known from western germany, netherlands, denmark, and scandinavia. mycenastrum corium (guersent) desv. the rather well documented history of retreat and advance of this conspicuous nitrophilous puffball was largely described and discussed by k r e i s e l (1982, maps), d ö r f e l t , b r e s i n s k y (2003, map), and b e n k e r t 2004 (maps). it extended from south-eastern europe or other continental regions to germany where it was found 1860 and 1889 near berlin, but then disappeared for several decenniums. from 1960 to 1992 many localities have been noted in most of eastern germany, but only one isolated locality could be recorded in western germany: 1982 near heidelberg (w i n t e r h o f f 2000), and a doubtful record from the fichtelgebirge mts. after 1992 m. corium again became rare in eastern germany, but in 2005 the species was still existent in a few localities near anklam, north-eastern germany. in poland, the species is known since 1890 and is registered from scattered places in southern and central poland (wo j e w o d a 2003). in scandinavia, m. corium was known as early as 1849 near malmö, and a series of localities became recorded in sweden from skåne through uppland till umeå and, frequently, on öland (l u n d q v i s t 1961; l a r s s o n 1997, map). there are a few records from south-western finland and åland islands (u l v i n e n in h a n s e n , k n u d s e n 1997). m. corium appeared 1961 on the danish island sprogø (hansen 1962) and 1976 on bornholm (b o r g e n 1977), and 1992 in southern norway (e c k b l a d 1992). in the netherlands, m. corium was observed 1976, 1977, and 1981 (a r n o l d s et al. 1995), and further records were from belgium and france. d ö r f e l t , b r e s i n s k y (2003) regard m. corium is an excellent indicator species for climatic changes (continentalization). a monitoring and continued mapping in far-reaching scale will be useful even in future. pleurotus eryngii (dc.: fr.) quél. is very rare north of its mediterranean and submediterranean main area. there are a few records from the netherlands (a r n o l d s et al. 1995), and two in rheinland-pfalz, western germany (k r i e g l s t e i n e r 1991, map). an ephemerous record in 1916 near elbląg (elbing) in northern poland is mentioned by wo j e w o d a (2003). there are no records from eastern germany, denmark, and scandinavia. polyporus rhizophilus pat. was known since 1930 in southern moravia, 1935 in slovakia, 1951 near prague and since 1962 in the bohemian middle mountains (š e b e k 1962, map; k o t l a b a 1984, map). 1961 it was found near mücheln in central germany (d ö r f e l t 1974, map), 1977 near bielinek nad odra (s k i r g i e ł ł o 1977), and more recently near pyrzyce in north-western poland and near przemysl (wo j e w o d a 2003). 1990 it was found near ribnitz in northeastern germany (j. d u t y unpubl.). there are no records from western germany, netherlands, denmark, and scandinavia. tulostoma giovanellae bres. [t. volvulatum borszcz. ss. hollós]. the mediterranean species appeared around 1900 near kecskemét and budapest, hungary (h o l l ó s 1904), later in wiener neustadt, austria (l o h w a g 1933), and again in budapest (b o h u s , b a b o s 1977), 1971–1974 in potsdam, eastern germany (k r e i s e l 1984). 88 h. kreisel all the central european occurrences had an ephemerous and synanthropic character, near house walls in towns. tulostoma pulchellum sacc. [t. hollosii z. moravec]. the species of wide continental distribution (it is known from north america as t. poculatum v. s. white, and originally from australia) was illustrated from hungary by h o l l ó s (1904, as t. fimbriatum) and is known from south-western slovakia and central bohemia (p o u z a r in p i l á t 1958). surprisingly it was collected 1967 1970 near berlin (k r e i s e l 2004). tulostoma squamosum j. f. gmel.: pers. the striking species has its main distribution in the mediterranean region and middle east, but has been collected in germany as early as in 19th century (1868, 1879) at several sites in berlin where it now is extinct. more recent records are from other regions of germany: 1956 ff. kyffhäuser mts. in central germany (r a u s c h e r t 1956), 1962 waren in north-eastern germany, before 1980 near bruchsal in the upper rhine valley and 1987 on the frisian island borkum (k r e i s e l 1984, map; krieglsteiner 1991, maps; w ö l d e c k e 1998). a record from poland was before 1991 near toruń (wo j e w o d a 2003), while the occurrence in scandinavia is doubtful. discussion since about 15 000 years, large areas in northern germany and northern poland, which had been covered by inland ice during the last (weichsel) glaciation of pleistocene, have been reoccupied by vegetation and by mycoflora insomuch every species of fungi which we actually observe in these areas, has reacted to a long-term global warming. it was the german microbiologist a u g u s t r i p p e l (in 1940, quoted in r i p p e l -b a l d e s 1955: 187) who assumed that the soil-inhabiting mould aspergillus niger tiegh. was spreading in germany from south to north during postglacial period. actually it is widely accepted that we are living in a period of accelerated warming, caused by human activities. it is to expect that this recent process is reflected also by changes in the mycoflora. if an organism seems to extend its distribution in a certain direction, two causes may be assumed: 1. the limits of distribution are really extending; 2. the knowledge of the organism is expanding from one region to the next one. which of these facilities is real in a special case, can be decided only by comparison of similar cases in neighboured regions. in case that parallel evolutions, or continued expansions, have been noted in various regions and countries, it can be concluded that the taxon really has extended its area. indroduction of additional organisms into a given territory may be due to two processes: either by anthopochory (neomycetes, as listed by k r e i s e l , s c h o l l e r 1994), often followed by a continuous expansion with broad front-line, or by spontaneous expansion, often marked at first only by unstable outpost-like occurrence (astathomycetes, k r e i s e l 2004). both kinds of processes may be combined, as described above in the cases of clathrus archeri, mutinus elegans, m. ravenelii, which have been introduced from other continents to europe, but have expanded continuously their distribution on this continent in the frame of their climatic ecological amplitudes. global warming and mycoflora in the baltic region 89 apart of anthropochory and climatic evolution, a third factor seems to have influence on expansion of fungal areas: the eutrophication, in particular with nitrogen, by excessive and careless enrichment of landscape with nutrients and fertilizers. mycenastrum corium is an excellent example for the changing frequency and distribution of a nitrophilous species in eastern germany in dependence of changing agricultural strategies. most of the fungal species, suggested in the statements above as examples for expansion from south to north as a consequence of global warming, are wood-destroying basidio mycetes (saprobes and tree parasites; section a). a lesser number of ectomycorrhizal basidiomycetes (section c), and at least one of the soil-inhabiting saprobes (agaricus bohusii, section b) can be added. in these categories, it seems that the greater river valleys with their floodplain forests and dunes (rhine, inn, elbe/łaba, oder/odra, and perhaps vistula) have much favoured the expansion process from south to north as can be noted in the cases of auricularia mesenterica, coriolopsis trogii, pluteus aurantiorugosus, agaricus bohusii, and chalciporus rubinus. category d, soil-inhabiting fungi of steppes and semide-serts, is the most problematic in this context, for the main direction of their expansion seems to be more from east and south-east to west, and anthropochory and eutrophication apparently play an important role in expansion. nevertheless, a “climatic continentalization” has been claimed as a factor for their unexpected and unstable appearence in central europe (d ö r f e l t , b r e s i n s k y 2003). strangely enough, no representant of ascomycetes could be included in one of the four sections. it may be due to their mode of spore dispersal and their diaspore survival during long-distance transports, that ascomycetes cannot respond to climatic changes in the same degree as basidiomycetes. acknowledgements: the author thanks for helpful suggestions and communications to dr. dieter benkert (potsdam), reinhard conrad (gera), christine morgner (bergen/vogtland), siegmund olm (neuenkirchen), dr. markus scholler (karlsruhe), dr. wolfgang tietze (zittau), and benno westphal (bobitz). references a n d e r s s o n h. 2004. internet. a n d e r s s o n o. 1941. bidrag til skånes flora 10. botaniska notiser 1941: 393 406. a r n o l d s e., k u y p e r t h . w., n o o r d e l o o s m. e. 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(in:) g. j. k r i e g l s t e i n e r (ed.). die großpilze baden württembergs 2. stuttgart, verlag eugen ulmer: 103 193. w o j e w o d a w. 1973. sarcodontia setosa (pers.) donk w polsce. fragm. flor. geobot. 19: 469 473. w o j e w o d a w. 2003. checklist of polish larger basidiomycetes. (in:) z . m i r e k (ed.). biodiversity of poland 7. w. szafer institute of botany, polish academy of sciences, kraków. 812 pp. w ö l d e c k e k. 1998. die großpilze niedersachsens und bremens. niedersächsisches landesamt für ökologie, hannover. 538 pp. global warming and mycoflora in the baltic region 93 globalne ocieplenie a mikoflora regionu bałtyku s t r e s z c z e n i e autor omawia możliwe skutki globalnego ocieplenia na rozmieszczenie i ekologię grzy bów makroskopowych oraz przedstawia przykłady sugerowanych gatunków wskaźnikowych, które są wyraźnie rozprzestrzenione od południa do północy. opracowanie dotyczy jedynie basidiomycetes, gdyż nie jest znany żaden przypadek niezlichenizowanych ascomycetes. au tor rekomenduje ciągły monitoring wymienionych gatunków. 2014-01-01t11:43:39+0100 polish botanical society the first locality of chalciporus rubinus (boletales, basidiomycota) in poland marek halama and jerzy szypuła museum of natural history, wrocław university sienkiewicza 21, pl-50-335 wrocław, marhalam@biol.uni.wroc.pl halama m., szypuła j.: the first locality of chalciporus rubinus (boletales, basidiomycota) in poland. acta mycol. 45(1): 57–65, 2010. chalciporus rubinus (w.g. sm.) singer, described in 1868 from england, was found in a city park in wrocław. this is the first record of the species from poland. macroand micromorphological characters of the polish specimens are described and illustrated. the delimitation of ch. rubinus, the knowledge of its distribution, ecology and conservation status is summarised. key words: chalciporus, rubinoboletus, urban greenery, wrocław, sw poland introduction the genus chalciporus bataille belongs to boletaceae, boletales, agaricomycetidae, agaricomycetes, basidiomycota, dikarya, fungi (šutara 2005; hibbett et al. 2007). it includes mycorrhizal, but usually not strictly specialized and not always obligatorily symbiotic fungi, occurring in temperate and tropical zones of both hemispheres. there are twenty three undoubtedly recognized chalciporus species worldwide and three of them occur in europe (klofac, krisai-greilhuber 2006; degreef, de kesel 2008; kirk et al. 2008; cf. singer 1986; horak 2005; muñoz 2005; šutara 2005). only one species of the genus, ch. piperatus (bull.: fr.) bataille (wojewoda 2003) has been reported from poland so far. the junior author found some fruiting bodies of an unusually coloured bolete species in the szczytnicki park, in the south-eastern part of wrocław (sw poland). the fungus was identified as chalciporus rubinus (w.g. sm.) singer, a species not yet known from poland. the aim of the paper is to describe the first collections of the species for poland, and to summarise the current knowledge of its taxonomy, distribution, ecology and conservation status. acta mycologica vol. 45 (1): 57–65 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 58 m. halama and j. szypuła material and methods material was collected on one locality. the description of macroscopic features is based on fresh material, on seven collections, comprising more than 40 basidiomata in all stages of development. the microcharacters of three recorded basidiomata (from three collections: nr 1, 2, 6) were observed and measured under a light microscope at magnification 1500× (basidiospores) and 800× (other features). for microscopic observations, dried pieces of basidiomata were placed in 5% nh4oh for about 5 minutes, then transferred to deionised water until they become pliable. free-hand sections of the rehydrated pieces of basidiomata were examined in 5%nh4oh, congo red and phloxine (in 1%nh4oh). amyloidity was tested with the melzer’s reagent. morphological measurements were made and are presented according to the method presented by breitenbach and kränzlin (1991). the abbreviation q is the ratio of basidiospore length to its width. terminology of morphological and anatomical elements has been adopted mainly from vellinga (1988). reported size of basidiospores, basidia (with sterigmata) and cystidia (cheilocystidia, pleurocystidia, caulocystidia), as well as dimensions of pileipellis hyphae were based on 31, 21, 31 and 31 measurements, respectively. basidiospore measurements, q coefficient and cystidia are presented as the mean, standard deviation, with the minimum and maximum dimensions in parentheses. dimensions of basidia are given as the range of minimum and maximum dimensions. drawings were made with the aid of a drawing tube under an oil-immersion objective. the voucher specimens of ch. rubinus have been deposited in the herbarium of the museum of natural history, wrocław university in wrocław, poland (wrsl). results chalciporus rubinus was found for the first time in poland on 12th of june 2007 in the szczytnicki park, in the south-eastern central part of wrocław (fig. 1). five carpophores growing on the ground in the neighbourhood of tilia and quercus were observed at the time. during the following forays made on 13th, 19th and 29th of june 2007 a few dozen of carpophores were discovered at the same locality and its nearest surroundings. on these occasions ch. rubinus was found under fagus, fraxinus, quercus, as well as philadelphus. later in august and september of the same year, the species was found at the same locality again. further observations confirming the occurrence of ch. rubinus at the investigated site were conducted in the period between july and september 2008. the location of the species has been carefully marked and will be monitored in future. chalciporus rubinus 59 description of the specimens chalciporus rubinus (w.g. sm.) singer figs 2–3 persoonia 7 (2): 319. 1973, (syn.: rubinoboletus rubinus (w.g. sm.) pilát & dermek, suillus rubinus (w.g. sm.)kuntze, xerocomus rubinus (w.g. sm.) a. pearson). macroscopic and microscopic characters. pileus 14-85 mm in diameter, hemispherical, subumbonate then plano-convex to even applanate and with reflexed margin; surface tomentose to finely fibrillose, dry and mat at first, lubricated and bright during wet weather, felted with age and even a little cracked during dry weather; brown, buff or alutaceous, usually keeping a pink tinge in about 5 mm from the margin. tubes first adnate, then slightly decurrent, up to 6 mm long, first pink or locally light yellow, then crimson-red to pink, the colour not changing in contact with air. pores irregular, angular, up to 1 mm in diameter, concolorous with the tubes, progressively becoming red-rose from the margin to the centre on age, the colour not changing under pressure. stipe 15-50×5-20 mm, solid, cylindrical with a tapering base or cylindrical, often slightly curved, smooth or somewhat ribbed by the decurrent tubes on the top, above concolorous with the pores, below red-rose to yellow, usually with chromeyellow basal mycelium. context in pileus and stipe quite compact, slightly watery in pileus, fibrillous in stipe, in pileus whitish with reddish patches, in stipe whitish to yellow and warm yellow at the base. smell indistinct or barely noticeable. taste mild. spore print pale brown. basidiospores (4.1) 6.3 ± 0.7 (7.2) × (3.8) 4.3 ± 0.5 (6.4) μm, q = (0.68) 1.47 ± 0.22 (1.76), ellipsoid to oblong, with a moderate hilar appendage, weakly pigmented, pale yellow (in 5%nh4oh), with a large guttula, inamyloid. basidia 30.4-43.3 × 9.3-11.7 μm, narrowly clavate, hyaline or containing small granules while immature, mostly with 4 sterigmata, without a basal clamp. cystidia (cheilocystidia, pleurocystidia) (36.6) 47.1 ± 5.9 (58.2) × (4.9) 7.1 ± 1.6 (9.8) μm, narrowly fig. 1. the location of the chalciporus rubinus site in wrocław (a) and poland (b; based on a 100 km atpol grid); 1 – urban boundary, 2 – municipal forests and parks, 3 – locality of the species. 60 m. halama and j. szypuła cylindrical to narrowly fusiform, potbellied, rarely narrowly clavate, erected or slightly curved, sometimes flexuose, dirty yellow (in 5%nh4oh), somewhat encrusted with crystalline bodies. caulocystidia (10.1) 27.0 ± 12.1 (53.5) × (2.2) 4.1 ± 1.4 (6.9) μm, cylindrical to clavate, usually articulate, mostly with obtuse apex, dirty yellow (in 5% nh4oh), accompanied by some scattered caulobasidia. pileipellis: a trichoderm transiting into a cutis, made up of usually interwoven, septate, cylindrical and thin-walled hyphae without clamps and with rounded, usually slightly broader (narrowly clavate to clavate) terminal elements (5,1-12) μm wide, with scattered incrustations and yellowbrown intracellular pigment (figs 2, 3). material examined. 1. wrocław (poland), szczytnicki park, soil humus, in the neighbourhood of tilia sp., quercus sp., 12.06.2007, leg. j. szypuła, wrsl; 2. wrocław (poland), szczytnicki park, soil humus, in the neighbourhood of fagus sylvatica, fraxinus sp., quercus sp., philadelphus sp., 13.06.2007, leg. m. halama, wrsl; 3. wrocław (poland), szczytnicki park, soil humus, in the neighbourhood of fagus sp., quercus sp., 19.06.2007, leg. j. szypuła, wrsl; 4. wrocław (poland), szczytnicki park, soil humus, in the neighbourhood of fagus sp., quercus sp., 29.06.2007, leg. j. szypuła, wrsl; 5. wrocław (poland), szczytnicki park, soil humus, in the neighbourhood of fagus sp., quercus sp., 29.08.2007, leg. j. szypuła, wrsl; 6. wrocław (poland), szczytnicki park, soil humus, in the neighbourhood of quercus sp. div., 21.07.2008, leg. m. halama, wrsl; 7. wrocław (poland), szczytnicki park, soil humus, in the neighbourhood of quercus sp., 20.09.2008, leg. j. szypuła, wrsl. taxonomical remarks. in september 1866, near dunstable (south bedfordshire, britain), worthington g. smith found plentiful fruiting bodies of previously unknown hymenomycetous fungus. it was clear to him, that observed basidiomata fig. 2. spores (a), basidia (b), cystidia (c) and caulocystidia (d) of chalciporus rubinus recorded in wrocław (12.06.2007, coll. by j. szypuła; drawn by m. halama). fig. 3. fruit-bodies of chalciporus rubinus recorded in wrocław (phot. m. halama, 21.07.2008). chalciporus rubinus 61 of the species belonged to the group ii, subtomentosi, of fries (1836-1838), and he described and illustrated the fungus under the name boletus rubinus (smith 1868). with improved knowledge of microcharacters and their application as generic criteria, the species have been placed in suillus gray (kuntze 1893-1898; singer 1965), xerocomus quél. (skirgiełło 1960), rubinoboletus pilát & dermek (pilát, dermek 1969) and chalciporus bataille (moser 1983; allesio 1985; singer 1986; horak 2005; muñoz 2005; klofac, krisai-greilhuber 2006). the approach of pilát and dermek (1969) was adopted by some authors of the most recent mycological papers (e.g., šutara 2005; knudsen, taylor 2008; šutara 2008). according to pilát and dermek (1969), the position of boletus rubinus w.g. sm. in such genera as xerocomus, suillus and chalciporus was isolated; the common feature of these genera is the fact that all their species have a uniform, elongate boletoid shape of spores. therefore, the transfer of this species to an independent genus, rubinoboletus, seemed to be a fairly acceptable solution (šutara 2005). nevertheless, the autonomous generic status of rubinoboletus is still controversial. šutara (2005) came to a conclusion that, with regards to the anatomical structure of the carpophores, chalciporus is very similar to rubinoboletus. he additionally found that it was very difficult to specify the boundary between the genus chalciporus and boletus subg. xerocomus (quél.) maubl. he gave the spore print colour as the main but rather doubtful criterion to distinguish between european representatives of chalciporus (cinnamon-brown or ferruginous-brown) and xerocomus (brownish with more or less strong olive tinges), and separated rubinoboletus on the basis of its short spores. the smaller spore size was not regarded as an important feature at the generic level by singer (1986). degreef and de kesel (2008) made a discovery of an interesting representative of chalciporus in africa. they described the species under the name chalciporus africanus j. degreef & de kesel and reported that it was similar to the temperate ch. rubinus (w.g. sm.) singer, but differed in its larger and more elongated spores, unchanging context and prominent reddish pileus colour. as a consequence, degreef and de kesel (2008) treated ch. africanus as taxon bridging the generic difference between chalciporus and rubinoboletus pilát & dermek. in their opinion the close relationship between ch. africanus and ch. rubinus confirms singer’s (1986) opinion that boletus rubinus w.g. sm. is undoubtedly a good species of chalciporus, making rubinoboletus a synonym of chalciporus. degreef and de kesel (2008) maintained that all globose-spored taxa subsequently combined in, or described under rubinoboletus should be placed elsewhere. moreover, they supported the corner’s (corner 1972 after, singer 1986) statement that “subglobose spores are to be expected in any alliance of elongated spores”. klofac and krisai-greilhuber (2006) took a similar approach; they proposed to include the genus rubinoboletus as a sub-genus in chalciporus. this point of view seems to be the most reasonable taxonomical concept and is also kept here. besides ch. rubinus, two other species of the genus occur in europe: ch. piperatus (bull.: fr.) bataille (syn. ch. hypochryseus (šutara) courtec)) and ch. amarellus (quél.) bataille (syn. ch. pseudorubinus (thirring) pilát & dermek; ch. pierrhuguesii (boud.) bataille) (klofac, krisai-greilhuber 2006). for identification, the basidiospores, the taste of flesh, the colour of tubes and pores are the most important 62 m. halama and j. szypuła features. ch. rubinus differs from other species of the genus mainly in the smaller size and broad elliptical to ovoid-spherical shape of basidiospores (q < 2). other significant features are not peppery (or even bitter) context, red-pink (reddish-ochraceous in age) pileus, carmine-red tubes and pores, red or carmine-red stipe with chrome-yellow base and occurring under deciduous trees (muñoz 2005). mild taste is also a specific feature of ch. amarellus, while the distinct peppery taste is a distinctive character of ch. piperatus. fruiting bodies of ch. amarellus are characterized by cream to pale orange-brownish or yellow-brownish (and sometimes pinkish-cream along the margin) pilei, pale yellow to intense yellow stipes and pink or reddish-pink (ochre with age) tubes and pores. the species is associated with abies, picea or pinus and has a tendency to occur at high altitudes in the mountains (gminder 1994; horak 2005; muñoz 2005). ch. piperatus, besides its distinct peppery taste, is characterized by dark red to rusty-brown pores, rusty-yellow to reddish-ochre pileus and concolorous, but normally lighter, bright yellow or chrome-yellow towards the base stipe (horak 2005; muñoz 2005). the species seems to be associated with coniferous (pinus, picea, abies) and deciduous trees (quercus, fagus, betula, castanea), but its mycorrhizal status is regarded as doubtful (högberg et al. 1996; knudsen, taylor 2008). some authors suspected ch. piperatus of an association with amanita muscaria (l.: fr.) lam. (spooner, roberts 2005). the relation of these species was confirmed by veerkamp and arnolds (2008), although the mechanism still has to be explained. habitat and distribution. carpophores of ch. rubinus appear in the summer and autumn (skirgiełło 1960; dermek, pilát 1991). the species is considered to be a mycorrhizal fungus (antonín et al. 2006; knudsen, taylor 2008), associated exclusively with deciduous trees (singer 1965). it was usually observed under quercus (singer 1965; allesio 1985; dermek, pilát 1991; hardtke, otto 1999; horak 2005; legon et al. 2005; muñoz 2005; antonín et al. 2006; arnolds, veerkamp 2008; knudsen, taylor 2008), but also the records from the neighbourhood of fagus, tilia, castanea, ilex and crataegus are known (michael, hennig 1971; muñoz 2005; b.m.s. 2009). everywhere in europe, ch. rubinus was generally recorded under isolated trees in old parks on alluvial riverside habitats (legon et al. 2005; kreisel 2006; arnolds, veerkamp 2008). moreover, it was also found in warmer deciduous and mixed forests, in roadside verges planted with trees, in gardens and on playing fields (skirgiełło 1960; antonín et al. 2006; b.m.s. 2009). ch. rubinus is known hitherto only from europe. in the 1960s it was considered to be a very rare taxon, known only from england, former czechoslovakia and germany (saxony) (singer 1965). nowadays the species is widespread but regionally usually treated as very rare (muñoz 2005; arnolds, veerkamp 2008). the present distribution area of ch. rubinus extends mainly throughout the west to central europe, but localities scattered in southern and northern parts of the continent are also known. it is known from england (legon et al. 2005), germany (kreisel 1987; kleine et al. 2004), the czech republic (polčák 2003; skála 2003; antonín et al. 2006), the netherlands (keizer 1995; beenen et al. 2002), belgium (van de kerckhove 2001; van de kerckhove, walleyn 2006), bulgaria (assyov, denchev 2004), slovakia (lizoň 2001), austria, hungary (allesio 1985; muñoz 2005) as well as italy (allesio 1985), spain (rubio et al. 2006) and norway (bendiksen et al. 1999). chalciporus rubinus 63 recently some authors have drawn mycologists’ attention to changes in the distribution of ch. rubinus. antonín et al. (2006) has reported the species as an example of spreading taxon, gradually increasing its previous limited distribution range in the czech republic. this tendency has also been confirmed by kreisel (2006) in germany. nevertheless, earlier recognized as a rare species, ch. rubinus was included in the red lists of the above and other european countries. in the czech republic, germany and bulgaria it is treated as an endangered species (benkert et al. 1992; gyosheva et al. 2000; lizoň 2001), in great britain, norway and slovakia it is regarded as vulnerable (bendiksen et al. 1999; lizoň 2001; legon et al. 2005), while in the netherlands as near threatened (arnolds, veerkamp 2008). for many years fungus forays have remained concerned with potentially rich habitats in order to record the widest range of species diversity. as a result, they focused on rural sites, leaving urban areas with only occasional recording. in order to find a new localities of ch. rubinus it is necessary to investigate man-made landscape. anthropogenic habitats may usually appear worthless for fungi, but actually they present a variety of challenges for studies. acknowledgements. we owe our sincere thanks to jérôme degreef, irmgard krisai-greilhuber, omer van de kerckhove and josef šutara for their kind help with completing the mycological literature. our gratitude is also extended to beata pokryszko for reading the mannuscript and correcting our english. finally, ananonymus reviewers are kindly acknowledgement for their valuable suggestions. references allesio c.l. 1985. boletus dill. ex l. 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(in:) r. walleyn, e. vandeven (eds). standaardlijst van basidiomycota en myxomycota van vlaanderen en het brussels gewest. rapport inbo.r.2006.27. instituut voor natuuren bosonderzoek, brussel, p. 28. veerkamp m., arnolds e. 2008. nieuwsbrief paddenstoelenmeetnet 9. coolia 53 (1): 97–108. vellinga e.c. 1988. glossary. (in:) c. bas, t. w. kuyper, m. e. noordeloos and e. c. vellinga (eds). flora agaricina neerlandica. critical monographs on families of agarics and boleti occurring in the netherlands. vol. 1. a.a. balkema publishers, rotterdam: 54–64. wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków, 812 pp. pierwsze w polsce stanowisko chalciporus rubinus (boletales, basidiomycota) streszczenie autorzy prezentują pierwsze w polsce stanowisko maślaczka rubinowego – chalciporus rubinus. owocniki tego gatunku, rosnące na ziemi, w sąsiedztwie różnych drzew liściastych (głównie dębów), zostały znalezione po raz pierwszy dnia 12 lipca 2007 roku w parku szczytnickim we wrocławiu. w pracy przedstawiono charakterystykę oraz zilustrowano najważniejsze cechy budowy makroskopowej i mikroskopowej znalezionych owocników, a także przybliżono taksonomię, ekologię i europejskie rozmieszczenie odnotowanego gatunku. 2014-01-01t11:50:25+0100 polish botanical society valsa viburni, a rare fungus in europe? vera hayova department of mycology, m.g. kholodny institute of botany tereschenkivska 2, ua-01601 kiev, vera.hayova@i.ua hayova v.: valsa viburni, a rare fungus in europe? acta mycol. 48 (2): 257–262, 2013. the paper provides brief illustrated description and general distribution of valsa viburni. the fungus is found to be highly host-specific and confined to viburnum lantana. according to currently available data on its distribution, the species has small number of records, fragmented range and is shown to be rare in europe. however, before assessment of the species, information on any additional unrecorded specimens is needed. on the example of v. viburni, some issues on fungal conservation for species of microfungi are considered. key words: valsa viburni, rare fungus, viburnum lantana, conservation of microfungi introduction the genus valsa fr. 1849 (valsaceae, diaporthales) comprises mostly corticolous fungi inhabiting various woody plants worldwide, often found in their cytospora anamorphic states (spielman 1985; castlebury et al. 2002; rossman et al. 2007). majority of valsa species have a wide host range and are associated with members of one or several plant families. some are known to occur on particular host genus, for example, on eucalyptus (adams et al. 2005). valsa viburni fuckel 1870 is apparently a strictly host-specific fungus confined to a single host species, viburnum lantana (adoxaceae). distribution of this fungus is likely much narrower than its host range. v. lantana, or wayfaring tree, is native to europe, northern africa, asia minor, caucasus and northwestern iran. the plant is also cultivated outside its native range. for example, it is introduced in north america where it has naturalized and reported invasive in some parts of usa (http://plants.usda.gov/). however, all but one of known up to now reports of valsa viburni are restricted to europe, except one record from africa, morocco. moreover, the species is mostly known by a single record in each country, except duplicates in original and exsiccate collections dated back to the 19th century and six recent records from four localitites in ukraine. acta mycologica vol. 48 (2): 257–262 2013 doi: 10.5586/am.2013.027 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 258 v. hayova thus the fungus appears to be rare. more information on its distinctive characters and current distribution is provided below. one of the main purposes of the paper is to draw attention of mycologists studying corticolous microfungi to this species which seems well defined and quite easily distinguished. the paper is also aimed to contribute to fungal conservation issues, particularly conservation of microfungi, using a single host-specific species as an example. materials and methods morphological description of anamorphic and teleomorphic states of v. viburni is mostly based on the specimens collected by author in 1983-1984 and 2008 in ukraine. these and additional specimens examined by the author are marked with an asterisk (*). analysis of general distribution of v. viburni is based on the data listed in the cited below bibliographic sources and databases available through the internet including gbif portal (http://data.gbif.org), usda fungal database (http://nt.arsgrin.gov/fungaldatabases), cybertruffle (www.cybertruffle.org.uk), etc. microscopic observations were made on dried material after mounting in distilled water or 1% cotton blue in lactophenol. measurements were done based on 20 dimensions of asci, spores and conidia in each of the studied specimens. photographs of general appearance were taken under carl zeiss stemi 2000-c stereomicroscope. microphotographs were taken under primo star microscope, canon a300 digital camera and axiovision 4.7 software, also used for measurements of microstructures. the author’s collections from ukraine are deposited in mycological herbarium of the m.g. kholodny institute of botany in kiev, ukraine (kw). results and discussion valsa viburni fuckel (fig. 1) jahrbücher des nassauischen vereins für naturkunde 23-24: 201 (1869-1870) [1870]. – engizostoma viburni (fuckel) kuntze, revisio generum plantarum 3 (2): 475 (1898) stromata scattered, subglobose to conical truncate, up to 2–2.5 mm diam. at the base, immersed in the bark and erumpent. ectostromatic disc prominent, whitish grey or ashy to olive, circular to oval, up to 1 mm diam. ectostroma inside light grey to brownish grey, surrounded from top to bottom by a thin blackish ventral zone gradually disappearing below. entostroma indistinct; sometimes a very fine blackishgrey layer below ascomata is present. ascomata perithecial, (3–) 8–12 (–19) in each stroma, arranged more or less in a circle, laterally inclined, black, globose or subglobose, with long ostiolar beaks thickened at the top. ostioles usually centrally inserted, appearing on the disc as a loose cluster, often slightly projecting asci 8-spored, clavate or subclavate, unitunicate, 54–68 (–77) × 8–10 μm, becoming detached at the base and freely floating, with inamyloid apical ring. ascospores allantoid, aseptate, colourless, (9.5–) 11–13 (–16.5) × (1.8–) 2.5–3 (–4.2) μm. paraphyses not observed. valsa viburni, a rare fungus in europe? 259 fig. 1. valsa viburni: a. stroma in surface view showing perithecial bumps and ectostromatic disc with a bunch of projecting ostioles, b. stroma in vertical section, c. ascus, d. ascospores, e. conidia, f. pycnidial stroma in surface view, with prominent ectostromatic disc and wide ostiole, g. vertical section of pycnidial stroma showing a blackish zone underneath, h. conidioma in transverse section. scale bars for a, f, g = 500 μm, for b = 250 μm, for c-e = 10 μm, for h = 300 μm. 260 v. hayova pycnidial stromata quite similar in shape to those of teleomorph, up to 1.5 mm diam. ectostromatal disc prominent, up to 500 μm diam., more or less circular, whitish or light-grey to ashy, with one, rarely 2 ostioles; ostiolar canal widening towards the top, ostiole black, rounded to elliptical or sometimes compressed, projecting or at the same level as disc surface. ectostroma greyish to dark grey inside, laterally surrounded by a thin blackened layer clearly visible in tangential sections through its upper part and gradually disappearing below. entostroma not distinct; ventral blackish zone occasionally present. conidiomata circular, elliptical or irregular, multi-chambered, comprising numerous radially arranged locules connected in the centre, each with additional invaginations. conidiophores unbranched or branched, colourless, 9–15 × 2 μm. conidiogenous cells cylindrical, slightly tapered at the apex, 6–8 (–11) × 1.5–2 μm, each with a minute collarette and a little periclinal thickening. conidia allantoid to nearly straight, aseptate, colourless, (3·6–) 4–4.8 (–5.2) × 1–1.2 μm. on corticated twigs and small branches of viburnum lantana l. geographical distribution. europe: austria, bulgaria, czech republic, germany, italy, romania, switzerland, ukraine. north africa: morocco. this distribution is based on the following specimens or bibliographic references: austria: v.1939 (petrak, 1940). bulgaria: 26.06.1989, v. fakirova (fakirova 2004; stoykov, denchev 2006; stoykov 2012). czech republic: 06.09.1957 (urban 1958). germany: 05.1893, j.n. schnabl (allescher & schnabl, fungi bavarici, no. 250), br 099228, 94*; 04.1896, j.n. schnabl (rabenhorst-pazschke, fungi europaei et extraeuropaei, no. 4163), br 099229,95*. italy: northern italy (saccardo 1882). morocco: 07.11.1960, r. bertault, mpu b03403. romania: 27.07.1964 (sandu-ville 1971). switzerland: 02.05.1871, p. morthier (herbier barbey-boissier, no. 231), br 099227,93*. ukraine: vinnytsia region, 15.07.1983; cherkasy region, 02.09.1984, v. hayova (merezhko, smyk 1991); khmelnytskyi region, 25.08.2008; v. hayova, kw 39968*, kw 39969*; 28.08.2008; v. hayova, kw 39970*, kw 39971*. v. viburni and its anamorph have several diagnostic features. externally, they have prominent greyish discs with either a projecting cluster of ascomatal ostioles, or a single conidiomatal ostiole widened at the top (fig. 1a, f, g). in addition, in both states ectostroma is encircled by a thin black line, often visible outside and particularly on transverse sections below ectostromatic discs, also noticeable on vertical sections (fig. 1b, f, g). another distinctive character, a blackish or dark grey ventral zone underneath stromata is often present (fig. 1g). more detailed description of morphological characters of this species as well as notes on its subgeneric position, connection with anamorph etc. is published separately (hayova, minter 2012). quite close to v. viburni is one of the most common and widespread valsa species, v. ambiens, occuring on numerous hosts including viburnum (e.g. v. opulus). v. viburni differs from v. ambiens by smaller ascospores and asci, general appearance of ostioles, ectostromatic disc and structure of ectostroma. due to scanty descriptions, these distinctions are usually not indicated resulting in misidentifications. for example, a report of v. viburni from sweden (ups bot f-133236, on viburnum opulus, k. holm & l. holm, 28.10.1984) is presumably erroneous and refers to v. ambiens. there are two other valsa taxa described on v. lantana: valsa perfodiens nitschke and v. opulina sacc. f. lantanae bres. the former species has much smaller microstructures, asci 28 x 4 μm and spores 8 x 2 μm (saccardo 1882); the latter form has similar dimensions of asci and spores to v. viburni (saccardo 1891). valsa viburni, a rare fungus in europe? 261 in our observations, anamorph and teleomorph usually develop in separate stro-namorph and teleomorph usually develop in separate stroand teleomorph usually develop in separate stromata. they are scattered and intermixed on the same twig, or occur on different twigs. the anamorph occasionally can be observed on dying plant parts and the fungus probably becomes saprobic later, when ascomata are produced. the teleomorph can be found both on attached and fallen twigs. it is entirely possible that like many other diaporthales members (rossman et al. 2007), v. viburni may colonize healthy twigs and remain there as a symptomless endophyte. unlike many widespread valsaceous fungi, this species is apparently rare on european continent. it is known as a single record in most of the listed above countries. records from three countries (switzerland from where it was described, italy and germany) are dated by the 19th century (fuckel 1870; saccardo 1882). other three specimens were collected around mid-20th century from austria, czech republic and romania (petrak 1940; urban 1958; sandu-ville 1971). one collection from the late 80s was reported from bulgaria (fakirova 2004; stoykov, denchev 2006; stoykov 2012). and the most recently, in ukraine where the species was specifically sought, only six records from four localities within quite restricted area in south-west of the country were generated. there might be several reasons for a small number of records of this fungus. first, as a species of microfungi it can be overlooked. however, v. viburni has quite large immersed stromata, usually conspicuous as bumps in the bark with prominent ectostromatic discs erumpent in the centre. second, this species can be underrecorded because of declining number of experts capable to observe and to identify it. still, many other species of valsa inhabiting various hosts are quite frequently reported. finally, so far it was found to occur exclusively on viburnum lantana. since the fungus is host-specific, it can be omitted if no attention is paid to the host plant during mycological observations. the host plant, viburnum lantana, occurs in woodlands, more frequently on their edges, and in open sites, particularly on calcareous soils in central, southern and western europe. v. lantana has not yet been assessed for the iucn red list (iucn 2012) but, from conservation point of view, most likely it could not be a cause for concern unless habitat destruction takes place regionally. apart from natural habitats, this shrub is also frequently planted as ornamentals in parks, gardens or hedges. however, in ornamental plantations diversity of corticolous ascomycetes can be rather low, mostly due to lack of dead twigs. in our observations, only one locality of v. viburni was in a neglected decorative plantation and three others were in natural habitats on the limestone hills. if the host plant was recognized as endangered, the associated fungus could be also regarded as a subject for conservation. however, v. lantana as well as the appropriate habitats do not seem to be rare. neverthless, v. viburni is known hitherto from a restricted number of records. thus, before any conclusion is made on how rare v. viburni is in europe, it is necessary to seek for any additional records of the fungus within this area. the highly host limited association, quite uncommon in the valsaceae family, simplifies searching for more localities of v. viburni. in the meanwhile, v. viburni can be assessed as data deficient using iucn red list categories and criteria (iucn 2001). and lastly, apart from teleomorph, it is important to take into account data on occurrence of the anamorph; for this purpose, distinctive morphological characters of both states are provided. 262 v. hayova conclusions this study demonstrates the major problem in evaluating conservation status of microfungi, i.e. uncertainty if the species is actually rare or simply rarely recorded. as the initial steps towards recognizing microfungi as conservation targets, fungal species confined to threatened hosts can be also recognized as rare. in case of v. viburni, a rarely recorded host-specific species is associated with rather common host plant; however, the fungus can be as well considered for conservation purpose if no more earlier records are found and no more successful observations on its occurrence in appropriate habitats are reported. regarding pleomorphic ascomycetous fungi, records of both states should be included in the species occurrence data. acknowledgments. the author is grateful to an anonymous reviewer for useful comments and suggestions on this manuscript. references castlebury l.a., rossman a.y., jaklitsch w.j., vasilyeva l.n. 2002. a preliminary overview of the diaporthales based on large subunit nuclear ribosomal dna sequences. mycologia 94:1017-1031. fakirova v.i. 2004. new records of bulgarian ascomycetes. mycologia balcanica 1: 41-43. fuckel k.w.g.l. 1869-1870 (publ. 1870). symbolae mycologicae. beiträge zur kenntniss der rheinischen pilze. jahrbücher des nassauischen vereins für naturkunde 23-24: 1-459. iucn 2001. iucn red list categories and criteria: version 3.1. iucn species survival commission. iucn, gland, switzerland and cambridge, uk. ii + 30 pp. iucn 2012. iucn red list of threatened species. version 2012.2. www.iucnredlist.org. downloaded on 21 december 2012. hayova v.p., minter d.w. 2012. valsa viburni. imi description sheets of fungi and bacteria, set 193, sheet 1928. merezhko t.a., smyk l.v. 1991. flora of the fungi of ukraine. diaporthales. kiev: naukova dumka, 216 pp. [in russian]. petrak f. 1940. beiträge zur kenntnis der pilzflora der umgebung von lunz am see und des dürrensteins in niederdonau. annales mycologici 38 (2-4): 121-180. rossman a.y., farr d.f., castlebury l.a. 2007. a review of the phylogeny and biology of the diaporthales. mycoscience 48: 135–144. http://dx.doi.org/10.1007/s10267-007-0347-7 saccardo p.a. 1882. sylloge fungorum 1: i-xix, 1-768. saccardo p.a. 1891. sylloge fungorum 9: i, 1-1141, 1891. sandu-ville c. 1971. ciuperci pyrenomycetes-sphaeriales din românia. editura academiei republicii socialiste românia, 409 pp. spielman l.j. 1985. a monograph of valsa on hardwoods in north america. canadian journal of botany 63 (8): 1355-1378. stoykov d.y. 2012. diaporthales. (in:) c.m. denchev (ed.). гъбите в българия. volume 8. institute of biodiversity and ecosystem research, bulgarian academy of sciences, sofia. 319 pp. stoykov d.y., denchev c.m. 2006. current knowledge of diaporthales (ascomycota) in bulgaria. mycologia balcanica 3: 179-185. urban z. 1958. revise československých zastupců rodů valsa, leucostoma a valsella. rozpravy československě akademie věd. 68 (12): 1-100. 2013-12-20t14:50:29+0100 polish botanical society laricifomes officinalis in the gorce mountains (s poland) władysław wojewoda department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków wojewoda w.: laricifomes officinalis in the gorce mountains (s poland). acta mycol. 45 (2): 129–131, 2010. information about second finding of laricifomes officinalis (batsch) kotl. & pouzar (fomitopsidaceae) in the gorce mountains in the external western carpathians is given, and its legitimate and illegitimate selected synonyms are cited. this species in poland is very rare and threatened. key words: polyporales, distribution, western carpathians, fungal conservation introduction laricifomes officinalis is very rare in poland (wojewoda, ławrynowicz 2006). historical localities of l. officinalis are cited by łuszczyński (2000) and chlebicki & łuszczyński (2002). the six contemporary localities were mentioned by piętka and szczepkowski (2004). in last years this species was published as fomitopsis officinalis (vill.) bondartsev & singer (e.g., ryvarden, gilbertson 1993; wojewoda 2003). taxonomy laricifomes officinalis (batsch) kotl. & pouzar basionym: boletus officinalis batsch, elenchus fungorum: 111. 1783 (legitimate). current name: laricifomes officinalis (batsch) kotl. & pouzar, česká mykol. 11(3): 158. 1957, legitimate. synonyms: boletus officinalis vill., histoire des plantes du dauphine 3: 1041. 1789, illegitimate; fomes officinalis (batsch) bres., iconographia mycologica 20: acta mycologica vol. 45 (2): 129–131 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 130 w. wojewoda 989. 1931, legitimate; fomitopsis officinalis (batsch) bondartsev & singer, ann. mycol. 39: 55. 1941, legitimate; agaricum officinale (batsch) donk, proceeding van de koninklijke nederlandse aka wetenschappen section c 74 (1): 26. 1974, illegitimate (mycobank 2009). classification according to kirk et al. (2008): fungi, basidiomycota r.t. moore (1980), agaricomycotina doweld (2001), agaricomycetes doweld (2001), agaricomycetidae parmasto (1986), polyporales gäum. (1926), fomitopsidaceae jülich (1982), laricifomes kotl. & pouzar (1957). distribution in the gorce mountains in the polish carpathians hitherto is known only one locality of laricifomes officinalis in krościenko nad dunajcem (skirgiełło 1959; wojewoda 1991). in the herbarium of the institute of botany, university of warsaw is preserved the one fragment of basidioma of fomes officinalis (vill.) neuman: krościenko, 1955, sept. 28, coll. a. skirgiełlo, det. h. orłoś, wa 7195. during the excursion in september 1955 in the vicinity of krościenko nad dunajcem (skirgiełło 1959, 2006) the fungi were collected in three geographical regions: the beskid sądecki mts, the gorce mts and the pie niny mts, but exact localities are not mentioned. according to skirgiełło (l.c.) most aphyllophoroid fungi were collected on the slope of ‘the elevation over zakrętki’ (it is the mount marszałek in the gorce mountains, wojewoda 1991). in 1970 laricifomes officinalis was found probably in the same locality: s poland, the external western carpathians, the gorce mountains, on s slope of the mount marszałek, under the peak of this mountain, ca 750 m a.s.l., in the larch forest with abies alba, fagus sylvatica and picea abies, ca 3 km n of krościenko nad dunajcem centre, on the trunk of old living larix decidua, 1970, april 17, coll. w. wojewoda, kram-f 48001 (fig. 1). it is necessary to search this species on the mount marszałek. acknowledgments. i should like to thank dr. maja graniszewska, the curator of the herbarium of the institute of botany, university of warsaw for loan the specimen of l. officinalis from krościenko nad dunajcem. references chlebicki a., łuszczyński j. 2002. fomitopsis officinalis (vill.: fr.) bondartsev & singer. atlas of the geographical distribution of fungi in poland 2: 61–67. kirk p. m., cannon p. f., minter d. w., stalpers j. a. 2008. ainsworth & bisby’s dictionary of fungi. 10. ed. cabi europe – uk. trowbridge, 771 pp. łuszczyński j. 2000. fomitopsis officinalis (coriolaceae) w polsce. fragm. flor. geobot. polonica 7: 271– 276. mycobank 2009. http://www.mycobank.org. piętka j., szczepkowski a. 2004. localities of fomitopsis officinalis in poland. acta mycol. 39 (1): 33–45. ryvarden l., gilbertson r. l. 1993. european polypores. 1. syn. fung. 6. fungiflora, oslo, 387 pp. skirgiełlo a. 1959. notatki mikologiczne z okolic krościenka nad dunajcem. monogr. bot. 8: 229–235. skirgiełlo a. 2006. zapiski ze stuletniego życia. bel studio sp., warszawa, 151 pp. fig. 1. laricifomes officinalis preserved in the herbarium kram f of the w. szafer institute of botany, polish academy of sciences in kraków. laricifomes officinalis 131 wojewoda w. 1991 (1990). pierwsza czerwona lista grzybów wielkoowocnikowych (macromycetes) zagrożonych w polskich karpatach. studia ośr. dokum. fizjogr. pan oddz. kraków 18: 239–261. wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland 7: 1–812. w. szafer institute of botany, polish academy of sciences, kraków. wojewoda w., ławrynowicz m. 2006. red list of the macrofungi in poland. (in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. 3. ed.: 53–70. w. szafer institute of botany, polish academy of sciences, kraków. laricifomes officinalis w gorcach (pd. polska) streszczenie we wrześniu 1955 roku a. skirgiełło zorganizowała wycieczkę mikologiczną w karpaty, do krościenka nad dunajcem. uczestnicy: a. skirgiełło, b. gumińska, a. nespiak i m. lisiewska (skirgiełło 1959, 2006) zbierali grzyby w trzech regionach fizyczno-geograficznych: w beskidzie sądeckim, gorcach i pieninach. dla okazu l. officinalis znajdującego się w zielniku zakładu systematyki i geografii roślin instytutu botaniki uniwersytetu warszawskiego jako stanowisko podane jest „krościenko” ale skirgiełlo (1959) informuje, że przeważającą liczbę polyporaceae zebrano na zboczu „wzniesienia nad zakrętkami”. wymienione „wzniesienie” to góra marszałek w gorcach, leżąca ponad krościenkiem nad dunajcem. uczestnicy wycieczki zbierali grzyby także na szczycie tej góry. w 1970 r. autor zebrał l. officinalis pod szczytem marszałka, być może na tym samym stanowisku, na którym gatunek ten znalazła a. skirgiełlo w 1955 r. w. wojewoda zebrał okazy tego gatunku 40 lat temu. nie wiadomo czy stanowisko to się zachowało. konieczne jest poszukiwanie na marszałku tego bardzo rzadkiego i zagrożonego w polsce grzyba. gatunek ten w ostatnich latach wymieniany był w publikacjach pod nazwą fomitopsis officinalis (vill.) bondartsev & sing. obecnie obowiązuje uprawniona nazwa laricifomes officinalis (batsch) kotl. & pouzar (mycobank 2009). 2014-01-01t11:50:53+0100 polish botanical society changes of hypogeous funga in the carpathian-pannonian region in the past centuries zoltán bratek, zsolt merényi and torda varga department of plant physiology and molecular plant biology, eötvös loránd university pázmány sétány 1/c, h-1117 budapest, bratek@caesar.elte.hu bratek z., merényi zs., varga t.: changes of hypogeous funga in the carpathian-pannonian region in the past centuries. acta mycol. 48 (1): 33–39, 2013. the exploration of hypogeous fungi in the carpathian-pannonian region speeded up in the past decades, owing to the widespread of truffle hunting with dogs. as a result, not only several new species were found in the region, but our view of the frequency of truffles also changed fundamentally. it became evident that tuber aestivum, t. brumale, t. macrosporum, t. magnatum, t. mesentericum and mattirolomyces terfezioides can be collected in commercial quantity. among the dog preferred hypogeous fungi (dph) several species, earlier believed to be rare like octaviania asterosperma and stephensia bombycina, also occurred. the taxonomic alterations and revisions brought about changes in the list of hypogeous fungi, and further changes are expected from molecular taxonomy research on a number of genera at present. key words: list of underground fungi, mycota, distribution, dog preferred hypogeous fungi, hungary introduction there are only few publications which have focused on the distribution of hypogeous fungi in european regions (ławrynowicz 1989, 1990, 1992; montecchi, sarasini, 2000; riousset et al. 2001). the carpathian-pannonian region is considered as one of the best known biogeographical regions of the hypogeous funga. this is partly because a number of manuscripts have reported and described hypogeous fungi since the 16th century, but mainly because two world-famous researchers, lászló hollós (1911) and lászló szemere (1965) made a detailed inventory of hypogeous fungi in the past nearly 200 years in their books. during his work hollós published roughly 460 data of 52 hypogeous fungi in the carpathian basin. szemere (1970) reported on 86 species in his book. such richness of the hypogeous funga may be attributed to the varied landscape, the superposition of several climatic effects and the diversity of acta mycologica vol. 48 (1): 33–39 2013 doi: 10.5586/am.2013.005 34 z. bratek et al. soil types in the carpathian-pannonian biogeographic region (stanners, bourdeau 1995). following the hardships of the last century, at the end of it, truffle hunting with dogs became popular in a number of countries in the region. the mapping and preservation of hypogeous fungi in fungaria also progressed by the leadership of professional organisations and societies. collections in the past decades did not only reveal species new to the region, but modified our knowledge on the frequency of occurrence and ecological requirements of some species. although the practice of using dogs for collection produced a large amount of data, it was evident that species with the most preferable odour were overrepresented. as collection for commercial purposes became general, collectors started to focus on certain forest types or landscapes, and dog owners often discouraged their dogs from finding non-commercial species. consequently, data from dog-aided collections can be considered representative only for species of commercial importance and their accompanying species. it is worth mentioning that these very data could have laid the foundations of the legal regulations for truffle collecting. the assessment of the distribution of other hypogeous species raises further questions in terms of sampling and collecting. being aware of the above mentioned diverse collecting methods and aims, in this paper we aim at reassessing the frequency of occurrence of hypogeous fungal species in the region and unrevealing the likely causes of changes and the possibilities of utilization and protection. materials and methods in cooperation with collectors of the first hungarian truffle society (emsze), we compiled a fungarium of 4224 preserved specimens of hypogeous fungi in the past three decades. data on collection and available habitat descriptions were organised in a database (merényi et al. 2010). present study compares the occurrence data of species in the database with the published data of hollós (1911) and szemere (1970). results and discussion table 1 shows the species used for comparison. some taxa in table 1 appear with genus names only as their classification at species level is still unclear, and they are under molecular taxonomic investigations. for simplicity, species will be discussed in three large groups. taxonomic changes and revisions. following the publication of szemere’s book (1970), many researchers revised his fungarium. a detailed evaluation of these revisions is given by bratek and halász (2005). the following species were removed from szemere’s list by the revisions: choiromyces magnusii (matt.) paoletti and endogone irregularis szem. new species on his list are: glomus caledonium (nicol. et gerd.) gerd. et trappe, glomus fasciculatum (thaxt.) gerd. et trappe and hymenogaster remyi dodge et zeller. all rhizopogon species were merged into rhizopogon roseolus changes of hypogeous funga in the carpathian-pannonian region 35 table 1 the occurrence data of taxons found in the carpathian-pannonian region based on the published data of hollós (1911), szemere (1970) and on the actual database of first hungarian truffle society (emsze) species hollós (1911) szemere (1970) emszze (2013) ascomycota elaphomyces aculeatus vittad. 8 2 26 elaphomyces anthracinus vittad. 5 3 2 elaphomyces asperulus vittad. 11 1 7 elaphomyces decipiens vittad. 6 elaphomyces granulatus fr. 9 31 elaphomyces leveillei tul. 1 2 elaphomyces maculatus vittad. 1 16 3 elaphomyces muricatus fr. 28 1 146 elaphomyces persoonii vittad. 4 elaphomyces reticulatus vittad. 10 elaphomyces virgatosporus holl. 11 balsamia platyspora bk. 2 balsamia polysperma vittad. 12 13 balsamia vulgaris vittad. 9 18 balsamia sp. (undet.) 32 hydnotrya cerebriformis harkn. 2 hydnotrya tulasnei berk. et br. 3 5 29 choiromyces meandriformis vittad. 84 77 genea hispidula berk. et br. 4 genea fragrans (wallroth) paoletti 11 genea klotzschii berk. & broome 3 36 genea lespiaultii corda 5 1 11 genea sphaerica tul. 1 15 genea verrucosa vittad. 2 3 115 genea sp. (undet.) 75 stephensia bombycina (vittad.) tul. 1 139 picoa carthusiana tul. 2 3 hydnobolites cerebriformis tul. 3 2 21 pachyphloeus spp. 3 13 95 mattirolomyces terfezioides (matt.) fisch. 1 7 45 tuber aestivum vittad. 51 6 461 tuber borchii aggr. (small white truffles) 20 6 227 tuber brumale vittad. 1 1 303 tuber excavatum aggr. 22 2 485 tuber lucidum bonn. 5 tuber macrosporum vittad. 2 146 tuber magnatum pico 50 tuber mesentericum (vittad.) fisch. 1 124 tuber nitidum vittad. 5 10 tuber regianum mont. et lazz. 9 tuber rufum pico 19 29 458 zygomycota endogone flammicorona trappe et gerd. 1 9 glomeromycota glomus caledonium (t.h. nicolson & gerd.) trappe & gerd. 1 glomus fasciculatum (thaxt.) gerd. et trappe 2 4 glomus fulvum (berk. & broome) trappe & gerd. 1 glomus macrocarpum tul. 6 10 glomus microcarpum tul. 6 6 basidiomycota leucogaster nudus (hazsl.) hollós 1 1 melanogaster ambiguus (vittad.) tul. 15 3 75 melanogaster broomeanus berk apud tul. 41 melanogaster intermedius berk. et br 2 36 z. bratek et al. (corda) t. m. fr. (martín, 1996). species new to the list due to rearrangements are: glomus macrocarpum tul. and glomus microcarpum tul. the revision of hymenogaster species by the complex evaluation of molecular and morphological data, and using samples from the carpathian-pannonian region revealed that hymenogaster griseus vittad. 1831 (emend stielow et al. 2010) includes the species h. lilacinus tul. (probably), h. lycoperdineus vittad., h. muticus berk. et br. (probably), h. populetorum tul. and h. vulgaris tul., while hymenogaster citrinus vittad. includes the species h. olivaceus vittad. preliminary molecular biology studies have indicated that revision of the traditional species boundaries is needed in some genera. the following genera are being revised on molecular taxonomical basis: arcangeliella-zelleromyces, gautieria, genea, glomus, hysterangium, melanogaster, pachyphloeus and sclerogaster. in other genera only certain species groups need to be revised by means of molecular taxonomy, such as elaphomyces muricatus/decipiens/reticulatus/asperulus, hymenogaster niveus aggr., hymenogaster rehsteineri aggr., tuber rufum aggr., tuber excavatum aggr. and tuber borchii aggr. new species, extinct species and very rare species. bratek et al. (1999) described sixteen species new to the carpathian-pannonian region including elaphomyces persoonii vittad., endogone flammicorona trappe et gerd., hysterangium pompholyx tul., sclerogaster compactus (tul.) sacc., tuber magnatum pico, tuber regianum mont. et lazz. and wakefieldia macrospora hawker. but for t. magnatum and melanogaster macrosporus velen. 4 melanogaster tuberiformis corda 2 melanogaster variegatus (vittad.) tul 22 2 12 melanogaster sp. (undet.) 49 rhizopogon roseolus sensu martín 15 10 57 rhizopogon villosulus zeller 2 octavianina asterosperma vittad. 6 38 phlyctospora fusca corda 1 3 16 gastroporium simplex matt. 2 7 hypogeic russulales 2 1 36 gautieria spp. 13 1 56 hysterangium calcareum hesse 2 hysterangium clathroides vittad. 21 2 hysterangium coriaceum hesse 3 hysterangium crassum (tul. & c. tul.) e. fisch. 27 hysterangium membranaceum vittad. 2 hysterangium nephriticum berk. 2 hysterangium pompholyx tul. 1 1 hysterangium stoloniferum tul. 11 2 29 hymenogaster tener berk. 3 4 3 hymenogaster arenarius tul. 2 7 8 hymenogaster bulliardii vittad. 35 hymenogaster citrinus vittad. 8 2 62 hymenogaster griseus vittad. 10 11 99 hymenogaster hessei soehner 28 hymenogaster luteus vittad. 1 103 hymenogaster niveus vittad. 41 hymenogaster rehsteineri bucholtz 23 wakefieldia macrospora hawker 2 sclerogaster spp. 14 table 1 – cont. changes of hypogeous funga in the carpathian-pannonian region 37 endogone flammicorona, which is often found in acidic conifer woodlands, all the above species are still regarded as very rare. t. magnatum has been collected in great quantities from the gallery forests of the southern pannonicum in the past few years. in a habitat rich in t. magnatum today, hollós used to collect other species. moreover, szemere collected plenty of hypogeous fungi in this county, too. all these facts may confirm the assumption that the distribution boundary of t. magnatum is shifting northwards as climate becomes warmer (bratek 2008). however, its advance is limited as the majority of riverine oak-elm-ash woodlands were cut and disappeared in the last century. during the above mentioned revision of hymenogaster, two species new to the region: h. megasporus soehner and h. pruinatus hesse, and two species new to science: h. intermedius stielow et al. 2010 and h. huthii stielow et al. 2010 were also found (stielow et al. 2011). stielow et al (2010) came across hydnotria michaelis (fischer) trappe, a species new to the region in sklene, slovakia. glejdura (2011) recorded stephanospora caroticolor, also new to the region in the lesser fatra (malá fatra), part of the northwestern carpathians. montecchi et sarasini (2000) reported on pachyphloeus prieguensis moreno-arroyo, gomez and calonge, also new to the region, in the vicinity of budapest. similarly, specimens of the species rhizopogon villosus zeller turned up in an arboretum near budapest (bratek 2006), which had probably got there together with plant specimens from america. the red list compiled by rimóczi et al. (1999), which assigns nature conservation values to macrofungi, contains most of the hypogeous fungi in hungary. according to this red list leucogaster nudus is critically endangered, while gastrosporium simplex and picoa carthusiana are endangered. our latest data confirm the validity of this categorization. of the native hypogeous fungi only six black-peridium elaphomyces species have so far received protection owing to their rarity (siller et al. 2005). e. pyriformis vittad. with also black peridium has not been found since hollós (1911) and is supposed to have vanished from the funga of the region. species with highly changed frequency of occurrence. table 1 contains a number of species whose frequency of collection has increased significantly or even by orders of magnitude. based on their pronounced, strong odour, it is fair to assume that the widespread of collecting with dogs is responsible for an increase in their frequency of collection. hence, we propose naming these species truffle dog preferred hypogeous fungi (dph). the number of fungarium data increased mostly for t. brumale, whose excellent odour places it among dph species. molecular studies have revealed genetic heterogeneity in the winter truffles of the region, confirming the hypothesis that t. brumale is not an invasive species in this biogeographical region, but has long been part of the hypogeous funga (merényi et al. 2012). the same stands for tuber mesentericum, whose genetic diversity has also been proved in the region (sica et al. 2007). tuber macrosporum also belongs here, as its rich localities have given the species commercial importance. we consider tuber aestivum a dph, too, since the number of its known localities has increased by more than one order of magnitude. the great majority of marketed t. aestivum originate from the high productivity woodlands of the jászság region, thus making hungary the most significant exporter of the species. 38 z. bratek et al. as of 2013, trufflers are obliged to keep records of the quantity of collected truffles in their collection notebook according to hungarian legal regulations, which enables forestry authorities to follow any changes in the amount of collected t. aestivum. an opposing trend, a significant drop of records occurs for choiromyces meandriformis. earlier, the species was common in local market places (hollós 1911), but has become rather unpopular in the european cuisine. at the same time, mattirolomyces terfezioides has the potential to become a rising star in truffle gastronomy thanks to its prominent sweet taste. besides a few records in other regions (kovács 2009), its richest habitats are found in the pannonicum, in black locust plantations growing on sand deposited by the danube (gógán et al. 2008). stephensia bombycina and octaviania asterosperma are also dph species that were classified as endangered due to the scarcity of data (rimóczi et al. 1999), but are not considered as rare anymore. in the above three chapters of the discussion, we presented species whose frequency of occurrence could be estimated from collection data. however, the unearthing of the hypogeous fungi of the carpathian-pannonian region is far from being finished, as some regions/landscapes have only slightly or not at all been studied. moreover, the mapping of surveyed species did not follow the most adequate method in all cases. the frequency estimation of dph species appears to be the most reliable/straightforward. the adequacy of mapping methodology has outstanding importance in the case of other species. a further obstacle to the evaluation of hypogeous funga is the uncertainty of species boundaries in several genera, where intensive molecular taxonomic work is needed or is in progress. the relevance of such research lies in the fact that ecological studies and conservation assessments can only operate with stable species concepts. all the above results have the potential to further clarify and harmonize our understanding of the funga of the carpathian-pannonian region and its changes. acknowledgements. this research has been supported by the mikoqual project under the ányos jedlik programme and by the qutaomel project under the national technology programme. we wish to thanks the reviewers for their valuable remarks and advice. references bratek z., albert l.i., bagi b., pálfy t., takács sz., rudnóy k., halász k. 1999. new and rare hypogeous fungi of carpathian basin. actes du ve congrès international, science et culture de la truffe et des autres champignons hypoges comestibles. 4-6 mars 1999, aix-en-provence, federation française des trufficulteurs: 255-256, france. bratek z., halász k. 2001. a kárpát-medence földalatti gombái. 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(eds). 1995. europe’s environment: the dobris assessment. – copenhagen: european environment agency. isbn 92-827-4713-1 stielow b., bubnek b., hensel g., münzenberger b., hoffmann p., klenk h.-p., göker m. 2010. “the neglected hypogeous fungus hydnotrya bailii soehner (1959) is a widespread sister taxon of hydnotrya tulasnei (berk.) berk. & broome (1846)”. mycological progress 9 (2): 195-203. stielow b., bratek z, orczán a.k., rudnóy sz., hensel g. 2011. species delimitation in taxonomically difficult fungi: the case of hymenogaster. plos one 6 (1): e15614. szemere l. 1965. die unterirdischen pilze des karpatenbeckens. akadémiai kiadó, budapest. szemere l. 1970. föld alatti gombavilág (the world of undeground fungi). mezőgazdasági kiadó, budapest. trappe j.m. 1969. comments on szemere’s „die unterirdischen pilze des karpatenbeckens”. mycologia 61: 170-174. http://dx.doi.org/10.2307%2f3757355 2013-06-22t22:39:53+0100 polish botanical society melampsoridium hiratsukanum – invasive rust species in lithuania, and its co-occurrence with eriophylid mite svetlana markovskaja laboratory of mycology, institute of botany, nature research centre, žaliųjų ežerų str. 49 lt-08406 vilnius 21, svetlana.markovskaja@botanika.lt. markovskaja s.: melampsoridium hiratsukanum – invasive rust species in lithuania, and its cooccurrence with eriophylid mite. acta mycol. 48 (2): 197–205, 2013. an invasive east asian rust fungus melampsoridium hiratsukanum, obligate biotroph belonging to pucciniastraceae, pucciniales (basidiomycota) is found widely spread on leaves of alnus incana in eastern, central and southern parts of lithuania. on alnus glutinosa this fungus is rare, sometimes occurring with an alder leaf pest, a microscopic eriophyid gall mite acalitus brevitarsus. information on the distribution patterns, ecological and morphological characters of this neomycete is given. key words: pucciniales, biotroph, melampsoridium hiratsukanum, alnus, acalitus brevitarsus, lithuania introduction majority of pathogenic neomycetes are classified as having considerable negative ecological and economic impact (pimentel et al. 2001; desprez-loustau et al. 2007, 2010; nobanis 2007). alien fungal diseases of woody plants represent a serious risk to forest ecosystems and growing plantations. in the mid-1990ies epidemic of a new foliar rust fungus affecting alder trees appeared in the baltic region, in estonia and finland (põldmaa 1997; kurkela et al. 1999) and started to spread rapidly in europe (hantula, scholler 2006). the agent was identified as melampsoridium hiratsukanum s. ito ex hirats. based on morphological characterization and later was confirmed by molecular data (hantula et al. 2009). in lithuania, the fungus appeared at the same time, the first specimen was collected in 1997 in ukmergė district on alnus incana (l.) moench (herbarium data, bilas 31446). originally m. hiratsukanum was described in 1927 on alnus hirsuta (spach.) rupr. in japan (hiratsuka 1927) and was reported as native species in far east asia (kuprevich, tranzschel 1957; hiratsuka et al. 1992; gjaerum 1996; chen 2002; cho, shin 2004; kobayashi 2007). m. hiratsukanum represents a heteroecious acta mycologica vol. 48 (2): 197–205 2013 doi: 10.5586/am.2013.021 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 198 s. markovskaja rust fungus belonging to pucciniastraceae (pucciniales, basidiomycota) characterized by macrocyclic host-alternating life mode. its uredinial and telial stages occur on alnus spp., while the aecial stage develops on larix spp. (kuprevicz, tranzschel 1957, kaneko, hiratsuka 1981; roll-hansen, roll-hansen 1981). the detailed comparison of urediospore morphology and r-dnr sequence analysis of the its region showed that european population of m. hiratsukanum is conspecific with population of this fungus from eastern asia and that both populations belong to a single palearctic population (hantula et al. 2009). in europe this aggressive rust commonly cause considerable damage to foliage of grey alder, but may also infect more resistant black alder and cause serious problems in forest stands mixed with larch (kurkela et al. 1999; hantula et al. 2009). introduction of this pathogen may be a result of an unintentional human activity such as transporting propagative plant material and seeds around the world or because of natural climatic factors. natural migration of m. hiratsukanum could be explained by continuity of its main (alnus spp.) and alternate (larix spp.) host distribution from the far east, where it inhabits native for asia manchurian alder (hiratsuka et al. 1992), to siberia (kuprevicz, tranzschel 1957), fennoscandia, baltic region and further to europe, where it occurrs on eurasian grey and native for europe black alders as well as on various (dahurian, siberian, european) larches (põldmaa 1997; kurkela et al. 1999; hantula et al. 2009; lilja et al. 2011). changing climate with milder winters and increased precipitation probably also stimulated invasion and rapid spread of this neomycete in the new regions on new hosts during the last decades. nowadays, m. hiratsukanum is widely spread in many european countries and is placed in the lists of important invasive species (hantula, scholler 2006; negrean, anastasiu 2006; nobanis 2007; desprez-loustau et al. 2007; desprez-loustau 2009, mułenko et al. 2010). the disease it causes is known from austria (riglerhager et al. 2003; kruse 2013), czech republic (dietrich 2005; müller 2003), estonia (põldmaa 1997), germany (scholler 1999; scholler et al. 2010; kruse 2013); hungary (szabo 2002), finland (kurkela et al. 1999, hantula et al. 2009 lilja et al. 2011), norway (gjaerum et al. 2004), poland (wołczańska 1999, piątek et al. 2001; mułenko et al. 2006, 2008, 2010), romania (negrean, anastasiu 2006), switzerland (meier et al. 2003), turkey (sert, sumbul 2005), italy (moricca, maresi 2010), uk (stringer 2010; hantula et al. 2012), ukraine (tykhonenko 2011) and from both americas (dennis 1970; gallegos, cummins 1981; ginns 1986; buritica, pardo cardona 1996; hernandez, hennen 2002; berndt 2004; hantula et al. 2012). the main aim of present study was to indicate the occurrence of melampsoridium hiratsukanum on alnus spp. in lithuania, to discuss its ecology and distribution patterns. material and methods samples of the rust infected leaves of alnus incana (l.) moench and a. glutinosa (l.) gaertn. were collected from various regions of lithuania during the autumn of 20092012. observation of disease symptoms (figs 1, 2) and severity continued until late october each year concerned. for comparison morphologically similar rusts, infected leaves of nearby growing betula pendula roth, which may occur on alder leaves as well, melampsoridium hiratsukanum – invasive rust species in lithuania 199 were collected. specimens with characteristic rust symptoms collected during present study and all specimens collected earlier by other mycologists available at the herbarium of the nature research centre, institute of botany (bilas), were microscopically examined using the standard microscopic techniques with a nikon stereo microscope at the magnifications up to 40x and with an olympus cx 41 microscope at the magnifications up to 400-1000x. the fungus was identified according to morphological characters using (kuprevich, tranzschel 1957; kaneko, hiratsuka 1981; roll-hansen, roll-hansen 1981; hiratsuka et al. 1992; hantula et al. 2009). description and illustrations were made from fresh preparations in distilled water. dried voucher specimens are preserved in the bilas herbarium. results and discussion on alders, three morphologically similar melampsoridium rusts may occur: m. betulinum (pers.) kleb , m. hiratsukanum s. ito ex hirats. and m. alni (thüm.) dietel, but only two of them – m. betulinum and m. hiratsukanum are known in europe so far (hantula et al. 2009). in lithuania, m. betulinum is widely spread on birches, but was not yet recorded on alders (minkevičius, ignatavičiūtė 1991). microscopical figs1-4. rust fungus melampsoridium hiratsukanum 1-2. symptoms on upper part of grey alder alnus incana -leaves (photo by j. kasparavičius). 3. melampsoridium hiratsukanum uredinium (scale bar = 100 μm). 4. ostiolar cells of uredinium and regularly echinulate urediniospores of melampsoridium hiratsukanum (scale bar = 20 μm). 200 s. markovskaja examination of lithuanian material showed that the rust developing on leaves of alnus incana and on leaves of a. glutinosa produced regularly echinulate yelloworange, ovoid to ellipsoidal urediniospores, commonly 24–32 × 12–15 μm in size with 4–6 bizonate germ pores and by size and morphology differs both from native rust m. betulinum and other asian alder rust, m. alni (tab. 1). following morphological characters of uredinia and urediospores the rust obtained on alnus leaves in lithuania was identified as m. hiratsukanum (figs 3-4). uredinia of m. hiratsukanum produced longer, resembling sharp spines, ostiolar cells (up to 50 μm), comparing to morphologically similar m. betulinum. uredinia of m. betulinum collected during present study were characterized by shorter ostiolar cells (up to 35 μm long) and by smooth, rounded upper end of urediniospores, which were slightly larger in size, commonly about 26–48 × 12–17 μm. m. alni is very similar to m. betulinum by size of urediniospores and lack of echinulation at the apex but its urediniospores have two germ pores, one at each end of the spore (hiratsuka 1927; kuprevich, tranzschel 1957; hantula et al. 2009). during present investigation, larix spp. trees were checked in various parks and forest plantations as well, but aecial stage of m. hiratsukanum was not found. it is possible that the fungus can reproduce and spread only by its urediniospores (hantula, scholler 2006). apparently, reduced fungus life cycle restricted to the telial and uredinal host (alnus spp.) prevails in lithuania. examined specimens: melampsoridium hiratsukanum: on alnus incana, gružu forest, ukmergė distr., 10 sep., 1997, leg. a.treigienė, det. ignatavičiūtė, bilas 31446; on a. incana, vilainiai forest kėdainiai distr., 16 sep., 1998, leg. a.treigienė, det. ignatavičiūtė; bilas 31445; on a. incana, plateliai forest, plungė distr., 3 sep., 2003, leg. a.treigienė, det. s. markovskaja, bilas 48792 on a. incana, semeliškiai environs, liaukiškiai forest, trakai distr., 6 sep., 2005, leg.a. treigienė, det. s. markovskaja, bilas 32914; on a. incana, vidiškiai forest, ignalina distr., 28 aug., 2009, leg./det. s. markovskaja, bilas 48794; on a. incana, biržai forest, biržai distr., 27 sep., 2009, leg./det. s. markovskaja, bilas 48795; on a. incana, sirvėtos forest, švenčionys distr., 29 aug., 2009, leg./det. s. markovskaja, bilas 48797; on table 1 comparison of urediniospores of m. hiratsukanum and similar rust species occurring on alnus and betula hosts from baltic countries and japan rust species host country urediniospores length and width range, μm m. hiratsukanum alnus incana lithuania 22.6-35.0 × 11.5-15.5 m. hiratsukanum alnus glutinosa lithuania 20.4-32.6 × 9.8-16.0 m. hiratsukanum alnus incana finland (kurkela et al. 1999) 19.2-34.5 × 10.6-19.6 m. hiratsukanum alnus glutinosa finland (kurkela et al. 1999) 22.6-35.2 × 10.9-16.6 m. hiratsukanum alnus incana estonia (kurkela et al. 1999) 17.9-30.9 × 10.6-17.3 m. hiratsukanum alnus glutinosa estonia (kurkela et al. 1999) 20.9-29.2 × 12.3-16.9 m. hiratsukanum alnus hirsuta japan (hantula et al. 2009) 17.0-32.6 × 8.8-19.7 m. alni alnus crispa japan (hantula et al. 2009) 24.4-43.3 × 9.1-16.4 m. betulinum betula pendula finland (kurkela et al. 1999) 26.5-42.1 × 11.6-19.9 m. betulinum betula pendula lithuania 28.0-42.5 × 11.5-18.4 m. betulinum betula pubescens lithuania (minkevičiūs, ignatavičiūtė 1991) (18)26. 2-32.1(50.0) × (10)12.7-13.9(17.0) m. betulinum betula pubescens finland (kurkela et al. 1999) 25.4-45.8 × 11.2-19.2 m. betulinum betula pubescens estonia (kurkela et al. 1999) 24.9-38.5 × 11.9-17.9 melampsoridium hiratsukanum – invasive rust species in lithuania 201 a. incana, labanoras forest, molėtai distr., 28 aug., 2009, leg./det. s. markovskaja, bilas 48798; on a. incana, pabradė environs, švenčionys distr., 29 aug., 2009, leg./ det. s. markovskaja, bilas 48796; on a. incana, curonian spit, neringa, smiltynė, 8 sep. 2009, leg./det. s. markovskaja, bilas 48791; m. hiratsukanum on alnus incana, vabalininkų environs, panevežys distr., 26 aug., 2009, leg./det. s. markovskaja, bilas 49521; on a. incana, petriškiai forest, širvintai distr., 10 oct., 2010, leg. a. treigienė, det. s. markovskaja, bilas 48932; on a. incana, jurkiškiai forest, molėtai distr., 22 sep., 2011, leg./det. s. markovskaja, bilas 50311; on a. incana, akmena environs, vilnius distr., 23 sep., 2011, leg./det. s. markovskaja, bilas 50309; on a. incana, antaviliai forest, vilnius distr., 5 sep., 2011, leg./det. s. markovskaja, bilas 50310; on a. incana, curonian spit, neringa, juodkrantė environs, 4 sep. 2012, leg./ det. s. markovskaja, bilas 50314; on a. incana, shoreline of curonian lagoon, klaipėda distr. 9 sep. 2012, leg./det. s. markovskaja, bilas 50312; on a. incana, aukštadvarys environs, trakai distr., 11 oct., 2012, leg./det. s. markovskaja, bilas 50316; on a. incana, rudninkai forest, šalčininkai distr., 21 aug., 2012, leg./det. s. markovskaja, bilas 50315; on a. incana, čepkeliai forest, varėna distr., 8 sep., 2012, leg./det. s. markovskaja, bilas 50317 on a. incana, kaunas environs, kaunas distr., 6 sep., 2012, leg./det. s. markovskaja, bilas 50318; on alnus glutinosa, together with acalitus brevitarsus fockeu, juodkrantė environs, neringa, curonian spit, 4 sep. 2010, leg./det. s. markovskaja, bilas 48933; on a. glutinosa, environs of lake tabaliukai, trakai district, 11 oct., 2012, leg./det. s. markovskaja, bilas 50313; melampsoridium betulinum: on betula pendula, kretinga environs, kretinga distr., 27 aug. 1971, leg. b. grigaliūnaitė, det. l. šidla, bilas 31443; m. betulinum on b. pendula, smiltynė forest, neringa, curonian spit, 9 sep. 2009, leg./det. a. treigienė, bilas 48458; m. betulinum on b. pendula smiltynė forest, neringa, curonian spit, 8 sep. 2011, leg./det. s.markovskaja, bilas 48459; m. betulinum on fig. 5. distribution map of melampsoridium hiratsukanum in lithuania (● on alnus incana, ■ on alnus glutinosa). the locality of the great cormorant colony (curonian spit, area in which m. hiratsukanum co-occurred with acalitus brevitarsus) is marked by ellipse. 202 s. markovskaja b. pendula, juodkrantė environs, neringa, curonian spit, 5 sep. 2012, leg./det. s. markovskaja, bilas 50319. distribution in lithuania. after inspecting mixed forest stands, parks and other habitats in various regions of lithuania, it was assessed that m. hiratsukanum is already widely spread in the country on alnus incana (leaves’ infection in some cases reached 20 to 80%) while on a. glutinosa it has appeared recently and does not cause strong damage of leaves (about 10%). most strongly infected (up to 60-80% of leaves) and defoliated already in september were juvenile grey alder trees growing along roads and water bodies. from 2009 till 2012, m. hiratsukanum was recorded on alnus incana in 22 localities of biržai, kaunas kėdainiai, klaipėda, kretinga, molėtai, ignalina, panevežys, plungė, šalčininkai, širvintai, švenčionys, ukmergė, trakai, and vilnius districts, while on alnus glutinosa it was found only twice, for the first time in 2010 at the edge of the great cormorant (phalacrocorax carbo sinensis) colony (curonian spit, environs of juodkrantė) and later, in 2012, in the environs of lake tabaliukai, trakai district (fig. 5). figs 6-9. acalitus brevitarsus. 6. deformation of alnus glutinosa leaves or blister-like galls and erineum (photo by r. iršėnaitė). 7. erineum clusters of hairs on an upper leaf surface (scale bar = 10 mm, photo by p. frey from lithuanian material). 8. an erineum hairs produced by leaf infested by mite (scale bar = 20 μm, photo by p. frey from lithuanian material). 9. microscopic mite acalitus brevitarsus (scale bar = 100 μm, photo by p. frey from lithuanian material). melampsoridium hiratsukanum – invasive rust species in lithuania 203 notable, that in curonian spit, in the forest stand affected by great cormorant colony, m. hiratsukanum infected leaves of both a. incana and a. glutinosa trees growing nearby and in some cases m. hiratsukanum infected alnus glutinosa leaves together with an alder pest, microscopic mite acalitus brevitarsus fockeu. symptoms caused on leaves by a. brevitarsus resemble rust fungus infection. this small arthropode, eriophyid mite (eriophyidae, acari, arthropoda) inhabit exclusively alnus glutinosa leaves (figs 6-9). like many gall-inducing mites, individuals of a. brevitarsus cause deformation of leaves or blister-like galls and elicit growth of clusters of hairs on upper surface of leaf (called an erineum) among which the mites live (figs 6, 7). the hairs have several near-horizontal branches at the tip (fig. 8), so each group of hairs is like a forest with a closed canopy. both the pest a. brevitarsus and the rust fungus m. hiratsukanum were recorded together on the same host leaves only in the stand affected by cormorants (curonian spit). in the other parts of curonian spit and in the continental part of lithuania they have never been found to occur together. evidently, the trees affected by cormorant excrements became less resistant to various pathogens and pests. successive infections of both agents can cause considerable damage to alder foliage and thus negatively influence the tree health and the functioning of the whole forest ecosystem. conclusion comparison of morphological data indicated that alders and birches in lithuania are infected by different rust species. leaves of alnus incana and a. glutinosa are damaged by invasive asian rust m. hiratsukanum, while native m. betulinum inhabits birch leaves. from its first record in 1997 till now m. hiratsukanum has widely spread in lithuania on a. incana. on a. glutinosa m. hiratsukanum was recorded for the first time in 2010, co-occurring with eriophyid mite acalitus brevitarsus. it is hypothesized that the successive colonization and co-occurrence of new pathogen (m. hiratsukanum) and pest (acalitus brevitarsus) on back alder was a result to the changes (decrease of trees resistance) caused by a strong negative impact of great cormorant colony, namely by forest hypertrophication from the bird excrements. acknowledgements. it is a great pleasure and honor to dedicate this paper to professor maria ławrynowicz. special thanks for help in identifying and making a photograph of the eriophyid gall mite acalitus brevitarsus are addressed to dr. pascal frey, and to my colleagues reda iršėnaitė and jonas kasparavičius for help during field trips. anonymous reviewers are 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jagiellonian university medyczna 9, pl-30-688 kraków, katarzyna.sulkowska-ziaja@uj.edu.pl sułkowska-ziaja k., muszyńska b., ekiert h.: chemical composition and cytotoxic activity of the polysaccharide fractions in sarcodon imbricatus (basidiomycota). acta mycol. 47 (1): 49–56, 2012. the aim of the study was chemical analysis of polysaccharide fractions from sporocarps of sarcodon imbricatus collected in natural sites and from the mycelium of in vitro cultures. three polysaccharide fractions (foi, foii, foiii) were isolated from sporocarps and two (fki, fkii) from in vitro cultures. qualitative analysis by hplc method showed that they are composed of galactose and fucose (foi, fki) or glucose and fucose (foii, fkii). foiii fraction of the sporocarps consisted of glucose only. molecular weights of isolated fractions ranged from 3.8 to 16.3 kda for fractions from the sporocarps and from 5.8 to 14.7 kda for that ones isolated from in vitro culture. the total percentage of sugar content for all fractions ranged from 97.8% to 99.1%. the percentage of uronic acids contents in acidic fractions was 2.6% and 2.7% for the foi and fki respectively. the work included also an assessment of cytotoxic activity of polysaccharide fractions in relation to tumor cell lines of human breast cancer mcv-7. foi polysaccharide fraction of the sporocarps inhibited the growth of cancer cells in 50% compared to the control at a concentration of 0.0125%, while the polysaccharide fraction fki from in vitro cultures inhibited cell growth in a concentration of 0.016%. key words: basidiomycota, polysaccharide fractions, cytotoxicity, breast cancer mcv-7 introduction basidiomycota species are an important source of biologically active compounds. substances with antiviral, antibacterial, cytotoxic and anticancer activity were detected in the sporocarps of this taxon (wasser 2002). the best-studied fungal metabolites, exhibiting therapeutic properties are polysaccharides. due this reason more and more studies are undertaken, not only on the chemical composition of the sporocarps but also in vitro mycelial cultures (sułkowska-ziaja et al. 2005). fungal polysaccharides include mostly acta mycologica vol. 47 (1): 49–56 2012 50 k. sułkowska-ziaja et al. glucans, mannans and galactans. biological activity is characteristic mainly for β-glucans in contrast to α-glucans which are rarely active and is determined by their chemical structure, particularly the type of glycoside bond (α or β) and spatial structure of polysaccharide molecule. polysaccharides with linear structure and without long side chains show the highest activity. it is connected with their better solubility in water and thereby easier bioavailability by the cells. therapeutically important polysaccharides have been found in microscopic as well as higher fungi. in clinical practice, amongst many pharmacologically active fungal polysaccharides, lentinan and krestin (so-called psk) are used (chihara et al. 1970). the object of our study was sarcodon imbricatus (l.: fr.) p. karst. occurring in spruce forests. this species is covered with strict protection in poland. the main goal of the study was examination of chemical composition and cytotoxic activity of the polysaccharide fractions from in vitro cultures and from sporocarps. material and methods samples of sporocarps. mature sporocarps of sarcodon imbricatus were collected in september 2008 in spruce forests in southern poland. after taxonomic identification according to hrouda (2005a, b) sporocarps were cut to small pieces and dried at 40ºc. in vitro culture of sarcodon imbricatus. initial cultures were derived from explants originated from the hymenial part of sporocarps which were sterilized with 70% ethyl alcohol and placed on petri dishes with solid medium according to lubiński with modifications (turło et al. 2004). cultures were incubated at a temperature 25±2ºc c under 12-h light (900 lx)/12 dark cycle and were subcultured every three week. experimental cultures were maintained as agitating ones in erlenmayer flask (500 ml) containing 250 ml medium under the same conditions as initial culture and were subcultured also every three weeks. isolation of crude polysaccharides. isolation of polysaccharides was performed according to mizuno method with modifications (mizuno 1999). dried sporocarps (50g) and lyophilized mycelium (25g) were refluxed with petroleum ether (1l) for 5 h to remove liposoluble constituents and then extracted with boiling water (1l) for next 5 h. the extraction process was repeated three times. the extracts were mixed, filtered, concentrated and centrifuged. the sevag method (darvill et al. 1985) was used to remove protein. obtained supernatant was added to absolute ethanol and kept overnight. the precipitate was collected and washed with absolute ethanol and acetone, then dried by lyophilization, yielding crude polysaccharide. fractionation of crude polysaccharides by ion-exchange column chromatography. crude polysaccharide fractions (1g) was dissolved in 100 ml of distilled water and loaded on deae-sephacel column. the column was at first washed with water, then with phosphate buffer ph 6.0 with increasing ionic strength and finally with aqueous naoh solution (0.2 m). two-milliliter fractions were collected. in order to detect polysaccharides, a 0.2 ml sample was taken from each eluted fraction that was chemical composition 51 mixed with sulfuric acid and phenol to yield color reaction (dubois et al. 1956). fractions containing polysaccharides were mixed, concentrated under vacuum, dialyzed and lyophilized. monosaccharide composition analysis. polysaccharide fractions (1g) were hydrolyzed in 1m trifluoroacetic acid for 10 h at 100oc in sealed glass tube. the monosaccharide compositions of the obtained solution were determined using hplc method. identification of the monosaccharides was carried out by comparing their retention times with those of standards under the same hplc conditions. briefly, the analytical conditions were as follows: hplc apparatus: type la chrom hitachi (merck); pump: l-7100; column: supelcosil lc-nh2 (250x4.6mm, 5μm); solvent system: acetonitryl : water 8:2 (v/v); flow rate: 1.3ml/min; detector: refractometric: l-74800; standards: l(+) arabinose, d(-) fructose, d(+) galactose, d(+) glucose, d(+) xylose, d(+) mannose, l(+) rhamnose (merck). determinations of moleculars weight. the molecular weight of polysaccharide fractions (1g) was determined by a gel filtration technique (rodriguezvanderwieles 1979). standard dextrans t-200, t-70, t-40, and t-10 were passed through a sepharose cl-4b column, and then the elution volumes were plotted against the logarithms of their respective molecular weights. determinations of the chemical character of the polysaccharide fractions. total sugar content in all the received fractions was determined by the phenol-sulfuric acid colorimetric method using glucose as the standard (dubois et al. 1956). total uronic acid content in all the received fractions was determined by m-hydroxydiphenyl method using galacturonic acid as the standard (filisetti-cozzi, carpita 1991). the examination of the cytotoxicity of polysaccharide fractions in the resazurin test. the influence of polysaccharide fractions on the metabolism of breast cancer tumor cells mcf-7 was examined with colorimetric method based on the reduction of the resazurin (sodium salt 10-oxide 7-hydroxy-3h-phenoxazin-3-one). resazurin is a metabolic activity indicator that in the oxidized form is a deep purple and when reduced turns to a light pink. spectrophotometric measurement of the absorbance for resazurin and resorufin performed at the wavelength λ=600 and 570 nm respectively (reddy et al. 1997). statistical analysis. obtained results were analyzed using non-parametric mannwhitney u test (n=3). differences with p<0.05 were considered to be statistically significant. the results were expressed as the mean values ± sd. results in vitro culture of sarcodon imbricatus. in agitated culture of s. imbricatus there was a 22-fold increase in fresh biomass within 3-week growth cycles. obtained biomass was used as a source of polysaccharide fractions. isolation and fractionation of polysaccharide fractions. during isolation process 15.1 g and 2.1 g of polysaccharides from the sporocarps and mycelium cultured in vitro were obtained respectively. efficiency of the isolation process in the case of sporocarps was 3.16%, while in the mycelium from in vitro cultures was 8.3%. 52 k. sułkowska-ziaja et al. fractionation of crude polysaccharide fractions was performed using deae ion exchange column chromatography. three polysaccharide fractions (foi, foii, foiii) were received from sporocarps and two (fki, fkii) from in vitro cultures. fractionation efficiency of the process amounted to 8.1%, 9.3%, 2.5% for the fraction of the sporocarps and 6.4% and 4.5% for the in vitro cultures. chemical analysis of polysaccharide fractions. monosaccharides composition, molecular weights, the total sugar and uronic acids contents of the polysaccharide fractions are presented in table1. qualitative analysis by hplc method showed that they are composed of galactose and fucose (foi, fki) or glucose and fucose (foii, fkii). fraction foiii consisted of only glucose. the average molecular weight of isolated fractions was estimated by reference to the calibration curve with standard dextrans and ranged from 3.8 to 16.3 kda for fractions from the sporocarps and from 5.8 to 14.7 kda for the polysaccharides isolated from in vitro culture. the total percentage of sugar content for all fractions ranged from 97.8% to 99.1%. the uronic acid contents were evaluated only in foi and fki 2.6 and 2.7% respectively. other examined fractions (foii, foiii and fkii) not revealed the presence of uronic acid. table 1 characteristic of polysaccharide fractions of sarcodon imbricatus polysaccharide fraction molecular weight [kda] monosacharide composition total sugars content [%] uronic acids [%] polysaccharide fractions from sporocarps foi 9.7±1.6 gal, fuc 99.8±1.8 27±1.5 foii 16.3±1.5 glu, fuc 99.1±1,6 -* foiii 3.8±0.9 glu 99.1±2.1 -* polysaccharide fractions from in vitro cultures fki 14.7±1.4 gal, fuc 98.5±2.2 2.8±0.8 fkii 5.8±1.5 glu, fuc 97.8±1.8 -* data presented as mean of 3 series ±sd; gal=galactose; glu=glucose; fuc=fucose; * not detected fig. 1. the degree of tumor cell inhibition of breast cancer lines mcv-7 by the polysaccharide fractions foi from sporocarps of sarcodon imbricatus. chemical composition 53 the study of cytotoxicity of the polysaccharide fraction against human tumor cells. assessment of cytotoxic activity of polysaccharide fractions in relation to tumor cell lines of human breast cancer mcv-7 was made. polysaccharide fraction foi from the sporocarps inhibited the growth of 50% cancer cells in comparison with the control at a concentration of 0.0125 % (fig. 1), while the polysaccharide fraction fki from in vitro cultures inhibited cell growth in a concentration of 0.016 % (fig. 2). other fractions not revealed antitumor activity in this test. statistical analysis revealed significant differences between control and all used concentrations (p<0.01) for fractions foi from sporocarps and for concentrations ranged from 0.01 to 0.04 for fraction fki from in vitro cultures. discussion one of the major groups of metabolites of medicinal importance are polysaccharides derived from basidiomycota (kohlmünzer et al. 1992; wasser 2002). polysaccharides from the sporocarps of sarcodon imbricatus were separated into three fractions: two neutral and one acidic, and from in vitro cultures into two: an acidic and neutral. all fractions were white, contained the 97.8 99.1% sugars. the polysaccharide fractions from sporocarps had higher total carbohydrate content and did not contain phosphorus, sulfur, nitrogen, protein or free amino acids. molecular weights of polysaccharides were ranged from 3.8 to 16.3 kda for sporocarps and from 5.8 to 14.7 kda for mycelial cultures. hplc analysis of acid hydrolysis products showed that they are composed of galactose, fucose and glucose. the obtained results indicated that glucose was the dominant monosaccharide in all the fractions. biologically active fungal polysaccharides are represented mainly by glucans, but also by galactans, mannans or fucogalactans (wasser, weis 1999). other studies on related species i.e. sarcodon aspratus revealed the presence of polysaccharide with structure known as fucogalactan (maruyama et al. 1989). fig. 2. the degree of tumor cell inhibition of breast cancer lines mcv-7 by the polysaccharide fractions fki from in vitro cultures of sarcodon imbricatus. 54 k. sułkowska-ziaja et al. fucogalactans are also present in sporocarps of other basidiomycota species e.g., coprinus comatus – (molecular weight of 1.03 kda) (fan et al. 2006) and sporocarps of ganoderma lucidum (molecular weight 2.8 kda) (bao et al. 2001; ye et al. 2008). the chemical composition of polysaccharides isolated from in vitro cultures is rare similar to polysaccharides isolated from sporocarps. a good example is a glucan from sporocarps of tylopilus felleus named tylopilan isolated at the department of pharmaceutical botany, jagiellonian university collegium medicum (defaye et al. 1988). tylopilan showed antitumor activity against transplantable sarcoma-180 cells in mice (more than 98% inhibition) and glioma cells (kohlmünzer et al. 1980 ). studies on the polysaccharides isolated from in vitro cultures of these species showed differences in their structure and biological properties. our biological studies have proved that both polysaccharide fractions from sarcodon imbricatus had inhibitory effects on breast cancer cells of mcv-7. polysaccharide fraction foi of sporocarps inhibited the growth of cancer cells in 50% compared to the control sample at a concentration of 0.0125%, while the polysaccharide fraction fki from in vitro cultures inhibited cell growth in a concentration of 0.016%. our earlier studies on the biological activity of polysaccharide fractions isolated from mycelium of sarcodon imbricatus cultured in vitro indicated very interesting antibacterial and antiviral (hpv) activity which prompts to continue studies of biological activity and also to compare the activity of fractions isolated from sporocarps (sułkowska-ziaja et al. 2011). acidic fraction contained galactose and fucose (fki) showed a higher microbial as well as cytotoxic activity. also the fraction from the sporocarps of a similar chemical nature showed cytotoxic activity (foi). literature data of genus sarcodon describe potential therapeutic effect of their metabolites and polysaccharide fractions can be considered as responsible for the cytotoxic activity. fucogalactan from sporocarps of sarcodon aspratus leads to the release of tumor necrosis factor-alpha (tnf-alpha) and nitric oxide in macrophages of mice in vitro. tnf-alpha production induced with 50 μg/ml of fucogalactan was significantly higher than induced by lentinan, the most active fungal polysaccharide with anticancer activity (mizuno et al. 2000). another example of a polysaccharide that shows cytotoxic activity against tumor cells sarcoma 180 is complex isolated from trametes versicolor (zjawiony 2004). in turn, polysaccharides isolated from pyrofomes demidoffi have cytotoxic influence against murine l929 fibroblastoma cells (zjawiony 2004). polysaccharides isolated from in vitro cultures of poria cocos are also good examples of cytotoxicity against the cells of sarcoma 180 in vivo (jin et al. 2003). conclusion the analysis performed during this study allowed to more specific identification of the metabolite composition in sporocarps of sarcodon imbricatus and determination of the biosynthetic abilities of the in vitro mycelium cultures. it has been demonstrated that in vitro cultures maintain ability to synthesize a range of metabolites occurring in the sporocarps like for example polysaccharides. results of the performed chemical composition 55 biological activity analyses lead to the conclusion that sarcodon imbricatus may be qualified as a species with potential therapeutic properties and the polysaccharide fractions isolated from both the sporocarps and in vitro cultures may be deemed as compounds responsible for therapeutic effects. the results of this study have not only cognitive importance but also have potential of practical application. it has demonstrated that in vitro cultures may be an alternative, high-yield source of a variety of biologically active metabolites. acknowledgements. the authors wish to thank professor florence caldefie-chezet from department of botany and microbiology faculty of pharmaceutical sciences at the university of clermont i, clermontferrand, france for the opportunity of testing cytotoxicity and agata piekoszewska for her help in 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(basidiomycota) cultured in vitro. acta pol. pharm. 68: 143–145. 56 k. sułkowska-ziaja et al. turlo j., lubiński o., gutkowska b. 2004. isolation of lentinan, an immunomodulating (1-3)-b-d-glucan from submerged cultivated mycelium of lentinus edodes and culture medium. acta pol. pharm. 61 suppl.: 40–42. wasser s.p. 2002. medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides. appl. microbiol. biotechnol. 60: 258–274. wasser s. p., weis a.l. 1999. therapeutic effects of substances occurring in higher basidiomycetes mushrooms: a modern perspective. crit. rev. immunol. 19: 65–96. ye l., zhang j., zhou k., yang y., zhou s., jia w., hao r., pan y. 2008. purification, nmr study and immunostimulating property of a fucogalactan from the fruiting bodies of ganoderma lucidum. planta med. 74: 1730–1734. zjawiony j. k. 2004. biologically active compounds from aphyllophorales (polypore) fungi. j. nat. prod. 67: 300–310. skład chemiczny i aktywność cytotoksyczna frakcji polisacharydowych sarcodon imbricatus (basidiomycota) streszczenie przedstawiciele gromady basidiomycota – której reprezentantem jest sarcodon imbricatus (l.: fr.) p. karst., są ważnym źródłem związków aktywnych biologicznie. w ich owocnikach wykryto m.in. związki o działaniu przeciwwirusowym, przeciwbakteryjnym, fungistatycznym, a także przeciwnowotworowym. najlepiej poznanymi metabolitami grzybowymi są polisacharydy. coraz częściej podejmowane są badania nad składem chemicznym nie tylko owocników, ale także grzybni pozyskanej z kultur in vitro. w badaniach z tego zakresu dowiedziono istnienia jakościowych i ilościowych różnic w produkcji niektórych grup związków chemicznych przez owocniki i grzybnię z kultur in vitro. obiektem przeprowadzonych badań były owocniki sarcodon imbricatus zebrane na terenie lasów świerkowych w roku 2008. z warstwy hymenialnej owocnika wyprowadzono kultury in vitro, które prowadzono, jako płynne, wytrząsane. w ramach przeprowadzonych badań wyizolowano 3 frakcje polisacharydowe (foi, foii, foiii) z owocników i 2 (fki, fkii) z mycelium z kultur in vitro. analiza jakościowa z wykorzystaniem metody hplc wykazała, że składają się one z galaktozy i fruktozy (foi, fki) oraz glukozy i fruktozy (foii, fkii). frakcja foiii z owocników składała się wyłącznie z glukozy. oznaczono masy cząsteczkowe wyizolowanych frakcji. wynosiły one od 3.8 do 16.3 kda dla frakcji z owocników i od 5.8 do 14.7 kda dla frakcji z kultur in vitro. całkowita procentowa zawartość cukrów dla wszystkich frakcji mieściła się w przedziale od 97.8% do 99.1%, natomiast procentowa zawartość kwasów uronowych we frakcjach o charakterze kwaśnym wynosiła 2.6% dla frakcji foi z owocników i 2.7% dla frakcji fki z kultur in vitro. druga część pracy obejmowała ocenę aktywności cytotoksycznej wybranych frakcji polisacharydowych w stosunku do linii komórek nowotworowych ludzkich raka sutka mcv-7 w teście in vitro. frakcje polisacharydowe foi i fki wykazywały działanie hamujące na komórki nowotworowe. frakcja polisacharydowa foi z owocników hamowała wzrost komórek nowotworowych w 50% w stosunku do próby kontrolnej w stężeniu 0.0125%, natomiast frakcja polisacharydowa fki z kultur in vitro hamowała wzrost komórek w stężeniu 0.016%. 2014-01-02t12:01:36+0100 polish botanical society auriculariopsis albomellea (agaricales, schizophyllaceae) new for poland władysław wojewoda bobrzeckiej 3/23, pl 31 216 kraków w o j e w o d a w.: auriculariopsis albomellea (agaricales, schizophyllaceae) new for poland. acta mycol. 41(1): 49 54, 2006. the article deals with the taxonomy, ecology, general distribution and threatened status of auriculariopsis albomellea bondartsev kotl. (basidiomycetes). in europe it is known only from czech republic, france, sweden and ukraine, in africa from canary islands, in north america from canada and united states. in poland the fungus was found for the first time in ne part of the country, in a pine forest, on dead twigs of pinus sylvestris. habitat and distribution of this saprobic fungus in africa, europe and north america are described, list of synonyms and important references are cited, polish name is proposed. key words: fungi, basidiomycetes, distribution, habitat, taxonomy, threat introduction in poland hitherto was known only one species from auriculariopsis genus: a. ampla (lév.) maire. it occurs especially on populus, also on salix, and is rather common in poland (wo j e w o d a 2003). in the fungarium of the institute of botany of the polish academy of sciences, was found second species from this genus: rare fungus – a. albomellea (bondartsev) kotl., new for poland. taxonomy cytidia albomellea bondartsev, bolezni rast. (morbi plant.) 16: 96.1927 (basionym). – cytidiella albomellea (bondartsev) parmasto, consp. syst. cortic. 101.1968. – auriculariopsis albomellea (bondartsev) kotl., česká mykol. 42(4): 239.1988. – phlebia albomellea (bondartsev) nakasone, mycologia 88(5): 766. 1996. cytidiella melzeri pouzar, česká mykol. 8(3): 127. 1954. – auriculariopsis melzeri (pouzar) stalpers, persoonia 13(4): 504. 1988. proposed polish name: uszaczek białobrzegi. systematic arrangement: basidiomycetes: aphyllophorales (nomen illeg. according to m i c h a e l et al. 1988), corticiaceae s. l. (d o n k 1964, j ü l i c h 1984, d o m a ń s k i acta mycologica vol. 41 (1): 49-54 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 50 w. wojewoda 1988, r o d r í g u e z a r m a s et al. 1992), lindtneriales, auriculariopsidaceae (j ü l i c h 1981), poriales, meruliaceae (m i c h a e l et al. 1988), stereales, meruliaceae (h a w s k w o r t h et al. 1995), schizophyllales, schizophyllaceae (k n u d s e n 1995, ve s t e r h o l t 1997), agaricales, schizophyllaceae (k i r k et al. 2001: 466, a k u l o v et al. 2003), corticiomycetes, corticiaceae, phlebioideae, merulieae (p a r m a s t o 1968, 1986). sometimes it was confused with auriculariops ampla, byssomerulius incarnatus, and stereum gausapatum (see n a k a s o n e 1996). descritpions and illustrations d o m a ń s k i (1988: 230, as cytidiella melzeri); e r i k s s o n , r y v a r d e n (1975: 339, figs 135-136, as cytidiella melzeri), j ü l i c h (1984: 158, as cytidiella melzeri), n a k a s o n e (1996: 767, figs 5, 9d-f, as phlebia albomellea), p o u z a r (1954: 126-127, figs without numbers, as cytidiella melzeri), ve s t e r h o l t (1997: 156, as cytidiella albomellea). for cultural descriptions see n a k a s o n e (1990). habitat and general distribution aurculariopsis albomellea occurs in forests and at skirts of peatbogs. basidiomata of this saprobic fungus occur on dead fallen trunks, and on attached or fallen corticate branches of coniferous and deciduous trees: abies, alnus, corylus, pinus and quercus, april–december. according to g i n n s and l e f e b v r e (1993) the fungus may be associated with a brown rot, according to n a k a s o n e (1996) it is associated with a white rot. for the first time it was found in sweden in 1905 (s t a l p e r s 1988). it is known hitherto from northern circumpolar: africa, europe and north america. africa: spain, canary islands: tenerife, in association with arbutus canariensis, picconia excelsa, and visnea mocanera, on dead wood, not identified (r o d r í g u e z a r m a s et al. 1992). europe: czech republic, bohemia, 5 localities: in forest with pinus and at a skirt of a peat-bog with ledum palustre, sphagnum sp., vaccinium myrtillus and v. uliginosum, on dead trunks and attached branches of pinus uncinata, on bark and wood of pinus sylvestris, and on dead, not fallen branches of quercus sp. (p o u z a r 1954, p i l á t 1969, n a k a s o n e 1996). france: on corylus avellana (b o i d i n , g i l l e s 1990); slovakia: on branch of pinus nigra (p o u z a r 1954); sweden: 2 localities, on pinus sylvestris, and on dead, dry branches of quercus robur (e r i k s s o n , r y v a r d e n 1975; s t a l p e r s 1988; n a k a s o n e 1996); ukraine: in forest, on dead branches of pinus sylvestris (b o n d a r t s e v 1927; k o t l a b a 1988; n a k a s o n e 1996; a k u l o v et al. 2003). north america: canada, yukon territory, on bark of fallen alnus crispa; united states: arizona, maine, mississippi, new mexico, wisconsin, on fallen or attached dead corticate branches of abies concolor, pinus palustris, p. ponderosa, p. resinosa and p. strobus (n a k a s o n e 1996). according to n a k a s o n e (1996, after h a l l e n b e r g 1981), auriculariopsis ampla is known also from iran in asia, but by h a l l e n b e r g (l.c.) this species is not mentioned. auriculariopsis albomellea 51 distribution and habitat in poland north-eastern poland: the niziny mazowiecko-podlaskie lowlands, the nizina północnomazowiecka lowland, the równina kurpiowska plain, the puszcza kurpiowska puszcza zielona forest, the puszcza myszyniecka forest northern part of the puszcza kurpiowska forest (k o n d r a c k i 2001), the mingos reserve (k o w a l s k a 1993); on some maps and in books as ‘mingus’ or ‘mirzgos’, 3.5 km nw of kuzie village, 27 km ne of ostrołęka, in pine forest with vaccinium vitis-idaea, on pure sandy soil (peucedano-pinetum sensu m a t u s z k i e w i c z 2001), on fallen dead corticate twigs of pinus sylvestris, 23 october 1976, leg. w. wojewoda (fig. 1). specimens examined kram f33108: basidiomata 0.2-2.1 x 0.2-1.1 cm in diameter, resupinate, at first regularly circular, orbicular to disc-shaped with loosening or incurved white wool margin, then sometimes confluent and some irregular, ceraceous when fresh, membranous when dry. outer surface white, tomentose. hymenophore smooth or some tuberculate, pale brown to brownish orange. hyphal system monomitic. hyphae 2.04.8 μm in diameter, hyaline, with thin or thick (up to 1.5 μm) walls. clamps at all septa of hyphae. cystidia none. basidia 28-42 x 4.5-7.0 μm, narrowly clavate, with subbasidial clamps, 4-spored. basidiospores 5.8-7.5 x 3.0-3.8 μm, narrowly ellipsoid, hyaline, smooth, thin-walled, non-amyloid (fig. 2). fig. 1. locality of auriculariopsis albomellea in poland. auriculariopsis albomellea 53 references a k u l o v a . y u . , u s i c h e n k o a . s . , l e o n t y e v d . v. , y u r c h e n k o e . o . , p r y d i u k m . p. 2003. annotated checklist of aphyllophoroid fungi of ukraine. mycena 2 (2): 1 75. a n o n y m o u s . 1995. red lists of macrofungi in the baltic and nordic region. expert seminar on threat ened species in the baltic region cryptogams, invertebrates, fish latvia, decemeber 4 8 1995, riga. working paper/draft, nov. 27, 1995: 1 58. b o i d i n j . , g i l l e s g . 1990. corticiés s.l. intéressants ou nouveaux pour la france (basidiomycotina). bull. soc. myc. fr. 106(4): 135 167. d o m a ń s k i s . 1988. corticiaceae acanthobasidium irpicodon. (in:) s . d o m a ń s k i (ed.). mała flora grzybów. 1 (5). basidiomycetes (podstawczaki) aphyllophorales (bezblaszkowce). państwowe wydawnic two naukowe, warszawa kraków, pp. 427. d o n k m . a . 1964. a conspectus of the families of aphyllophorales. persoonia 3 (2): 199 324. e r i k s s o n j . , r y v a r d e n l . 1975. the corticiaceae of north europe. 3. fungiflora. oslo, pp. 285 546. g ä r d e n f o r s u . (ed.). 2005. rödlistade arter i sverige 2005. the 2005 red list of swedish species. artdatabanken. swedish species information centre in cooperation with swedish environmetal pro tection agency, uppsala, pp. 496. g i n n s j . , l e f e b v r e m . n . l . 1993. lignicolous corticioid fungi (basidiomycota) of north america. systematics, distribution, and ecology. mycologia memoir 19: 1 247. h a l l e n b e r g n . 1981. synopsis of wood inhabitating aphyllophorales (basidiomycetes) and heterobasid iomycetes from n. iran. mycotaxon 12 (2): 473 502. h a w k s w o r t h d . l . , k i r k p. m . , s u t t o n b . c . , p e g l e r d . n . 1995. ainsworth and bisby’s dictionary of the fungi. 8 ed. imi, univ. press, cambridge, pp. 404. j ü l i c h w. 1981. higher taxa of basidiomycetes. bibl. mycol. 85: 1 485. j ü l i c h w. 1984. die nichtblätterpilze, gallerpilze und bauchpilze. aphyllophorales, heterobasidiomycetes, gastromycetes. (in:) h . g a m s (ed.). kleine kryptogamenflora. ii b/1. basidiomyceten. 1. g. fischer verl., stuttgart new york, pp. 626. k i r k m. p., d a v i d p. f., s t a l p e r s j. c. 2001. ainsworth & bisby’s dictionary of the fungi. 9th ed. cab international, wallinford, pp. 655. k n u d s e n h . 1995. taxonomy of the basidomycetes in nordic macromycetes. symb. bot. ups. 30 (3): 169 208. k o n d r a c k i j . 2001. geografia regionalna polski. wyd. 2. wydawnictwo naukowe pwn, warszawa, pp. 441. k o t l a b a f. 1988. správné jméno pro cytidiella melzeri pouz. česká mykol. 42(4): 239. k o w a l s k a e . (ed.). 1993. polska mapa ochrony przyrody. polskie przedsiębiorstwo wydawnictw karto graficznych im. e. romera. warszawa wrocław. m a t u s z k i e w i c z w. 2001. przewodnik do oznaczania zbiorowisk roślinnych polski. wydawnictwo na ukowe pwn, warszawa, pp. 536. m i c h a e l e . , h e n n i g b . , k r e i s e l h . 1988. handbuch für pilzfreunde. 6. die gattungen der großpilze europas. bestimmungsschlüssel und gesamtregister der bände i bis v. ed. 2. veb g. fischer verlag, jena, pp. 310. n a k a s o n e k . k . 1990. cultural studies and identification of wood inhabiting corticiaceae and selected hymenomycetes from north america. mycologia mem. 15: 1 412, n a k a s o n e k . k . 1996. morphological and molecular studies on auriculariopsis albomellea and phlebia albida and a reassessment of a. ampla. mycologia 88(5): 762 775. p a r m a s t o e . 1968. conspectus systematis corticiacearum. inst. zool. bot. acad. sci. ssr estoniae. tartu, pp. 361. p a r m a s t o e . 1986. on the origin of the hymenomycetes (what are corticioid fungi?). windahlia 16: 3 19. p i l á t a . 1969. houby československa ve svém životním prostředí. academia nakladelství československé akademie věd, praha, pp. 133. p o u z a r z . 1954. cytidiella melzeri g. n. et sp. n., nový typ resupinátních hub číšovcovitých. česká mykol. 8 (3): 125 129. 54 w. wojewoda r o d r í g u e z a r m a s j . l . , r y v a r d e n l . , h a l l e n b e r g n . , b e l t r á n te j e r a e . 1992. new and noteworthy species of aphyllophorales (basidiomycotina) from the canary islands. mycotaxon 45: 433 437. s t a l p e r s j . a . 1988. auriculariopsis and the schizophyllales. persoonia 13(4): 495 504. ve s t e r h o l t j . 1997. cytidiella pouzar. (in:) l . h a n s e n & h . k n u d s e n (eds). nordic macromy cetes. 3. heterobasidioid, aphyllophoroid and gastromycetoid basidiomycetes. nordsvamp. copenha gen, pp. 156. w o j e w o d a w. 2003. checklist of polish larger basidiomycetes. (in:) z . m i r e k (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków, pp. 812. auriculariopsis albomella (agaricales, schizophyllaceae) nowy gatunek dla polski s t r e s z c z e n i e w fungarium instytutu botaniki im. w. szafera pan w krakowie stwierdzono okazy auriculariopsis albomellea, gatunku ostatnio zaliczanego do rodziny schizophyllaceae w rzę dzie agaricales (basidiomycetes). okazy tego grzyba zebrano w 1976 r., w północno wschod niej polsce, w puszczy myszynieckiej (północna część puszczy kurpiowskiej), w rezerwacie mingos, w subkontynentalnym borze świeżym peucedano pinetum, na martwych, opadłych, pokrytych korą gałązkach pinus sylvestris. jest to gatunek nowy dla mikobioty polski. publiko wany był z europy (republika czeska, francja, słowacja, szwecja, ukraina), z afryki (hisz pańskie wyspy kanaryjskie) i z ameryki północnej (kanada, stany zjednoczone). rozpostar te, dyskowate, brązowawe, białoobrzeżone owocniki tego saprobowego grzyba z gładkim lub gruzełkowatym hymenoforem, występują na martwym drewnie drzew iglastych i liściastych: abies concolor, corylus avellana, pinus nigra, p. palustris, p. ponderosa, p. resinosa, p. sylvestris, p. strobus, p. uncinata, quercus robur i q. sp. uszaczek białobrzegi, jest gatunkiem rzadkim. w szwecji umieszczono go na czerwonej liście grzybów zagrożonych w tym kraju, z kategorią „lokalnie wymarły”. w polsce gdzie stwierdzono go tylko raz, w rezerwacie leśnym, też przy puszczalnie jest zagrożony. 2014-01-01t11:43:26+0100 polish botanical society immunomodulating and anticancer properties of fungi kazimierz kopczyński jan kochanowski university in kielce, branch in piotrków trybunalski słowackiego 114/118, pl-97-300 piotrków tryb., inp@unipt.pl kopczyński k.: immunomodulating and anticancer properties of fungi. acta mycol. 47 (1): 91–96, 2012. fungi contain a number of biologically active substances whose importance for human health has been confirmed in several studies. in particular, β-glucans, selenium, vitamin d, c and e should be mentioned. these substances play an important role in shaping the immune system and prevent cancer. β-glucans reduce the risk of cardiovascular disease and lower the cholesterol level. key words: selenium, β-glucans, vitamins, immunity, cancer introduction this article is a review and an attempt to synthesize the results of the studies on immunomodulating and anticancer properties of mushrooms. usually, scientific reports present only individual components being of importance for the human health contained in mushrooms (beuth, drebing 2006; marley 2009). however, such substances as β-glucans, selenium and some vitamins and minerals occur in mushrooms simultaneously, making the effect of stimulation of the immune system and anticancer action stronger. worth mentioning is the fact that glucans can be present in a wide variety of configurations, that have different biological activity. according to some researchers, the use of a mixture of different species of fungi with a wide variety of glucans can further enhance their immune effects. the study presents the health effects of substances contained in mushrooms, such as selenium, β-glucans, some vitamins and minerals. the main purpose was to expose the fact that these components appear jointly in fungi and their immunomodulating and anticancer activity has been confirmed by many studies. in the light of these studies, a widespread belief that mushroom’s values are only a taste and aromatic qualities seems to be a myth. the therapeutic value of many edible mushrooms, examples of which are given in this paper, has been thoroughly confirmed in the recent acta mycologica vol. 47 (1): 91–96 2012 92 k. kopczyński years, as indicated by the cited literature, including scientific reports in the field of oncology and immunology. this paper demonstrates practical importance of fungi in shaping the immune system and in preventing and fighting cancer. active compounds and their effects selenium is one of indispensable microelements for proper functioning of enzymatic systems. the principal function of selenium is creation of glutathione peroxidase, a powerful antioxidant enzyme. it protects red blood cells and cellular membranes from harmful effect of (free) radicals. it is also important for proper functioning of immunity system and thyroid gland. numerous research confirm that selenium decreases the risk of occurrence of all kinds of cancer, in particular liver, prostate, colon and lungs cancers (hasik 2000; beuth, drebing 2006; juchimiuk et al. 2010). when compared to the control group, significantly lower level of selenium in the serum was found among the patients with early and advanced stage of stomach cancer (juchimiuk et al. 2010). other data confirm that fewer cases of bronchial cancer were found in the group of patients with high level of selenium in serum (hasik 2000). also the risk of prostate cancer is four to five times higher in the group of patients with low level of this element in serum (brooks et al. 2001; rostock, saller 2008). selenium can mitigate effects of chemotherapy and radiotherapy, prevents the phenomenon of metastasis and protects against the ultraviolet radiation (beuth, drebing 2006). selenium present in the structure of glutathione peroxidase takes part in regeneration of vitamin e in biological systems (kostogrys 2007). vitamin e together with ß-carotene and selenium are among the most important antioxidants. it is belived that vitamin e is effective in high oxygene pressure conditions which can be found in lungs (szponar, respondek 2000). according to the literature, therapeutic substances that have significant anticancer effect contained in vegetables and fruit are described as blocking agents or supperssing agents depending on the strength and phase of their action. similarly, selenium is considered as a suppressing agent – acting through enzymatic breakdown of carcinogens (hasik 2000). the level of selenium in fungi varies within the limits from 0.31 to 19.86 mg/ kg d.m. its content depends not only on the species but also on the location and collecting season. it may also depend on the method of measurement of this element. results obtained with inductively-coupled plasma atomic emission spectroscopy (icp-aes) may differ from those obtained by other techniques. however, as noted by falandysz (2011) selenium is one of the chemical elements always present in mushrooms. the highest content of selenium was found in the following species of edible fungi: boletus edulis bull, xerocomus badius (fr.) j.-e. gilbert, stropharia rugosoannulata farl. ex murril, leccinum aurantiacum (bull.) grey (lasota, kalinowski and florczak 1994). immunomodulating properties 93 β-glucan is the polysaccharide extracted from the cellular membranes of yeasts (saccharomyces cerevisiae meyen ex e. c. hansen) a patented method of “gentle” extraction allows to preserve intact the bindings 1,3/1,6 d, which activate macrophages. β-glucans are strong stimulants of macrophages, increase immunity against different bacterial, viral, fungal and parasitic infections, delay ageing processes, slow down the growth of tumors, act as strengtheners of proliferation phase of wounds healing, protect against negative effects of standard cancer treatment, improve its effectiveness, prevent the phenomenon of metastasis, decrease the risk of circulatory system diseases, lower the cholesterol level and influence the antihyperglicemic effect (ber, gazella 2002; marley 2009; strach 2011). glucans are present in incredibly wide range of configurations that influence their biological activity. the ones with branches β-1,3 and β-1,6 proved to be the most active in immunity stimulation. overall growth of branching in size and complexity is accompanied by increase of biological activity (ohno 2005). according to some researchers, using a mixture of different species of fungi with a wide variety of glucans may cause an increased immune activity (marley 2009). glucans are not readily available without prior boiling of fungi in order to break the structure of their cell walls. cooking of the mushrooms enables digesting and releases the polysaccharides, which would otherwise remain in the indigestible cell structures (marley 2009). β-glucans and other polysaccharides with non-specific immunomodulational properties were found in such species of fungi as: pleurotus ostreatus (jacq.) p. kumm., tricholoma caligatum (viv.) ricken, auricularia auricula-judae (bull.) quél., lepista flaccida (sowerby) pat., sparassis crispa (wulf.) fr., lyophyllum decastes (fr.) singer (żurowska 2010). all these species are edible (lepista flaccida, a very spicy mushroom, can be consumed only in small amounts in addition to spicy roast). research conducted in national cancer institute of tokyo on the species flammulina velutipes (curtis) singer, demonstrated that it also contains polysaccharides and antioxidants: vitamin c and e (babal 2011). this species is especially valuable, since it occurs in winter, when no other edible mushrooms do. some authors indicate a large content of ß-glucans in mushrooms (agaricus) and in shiitake lentinus edodes (berk.) singer (ley 2008; babal 2011). however, there are reports on cancerogenous and mutagenous effects of these fungi as well (sadowska et al. 2004; škubla 2005). experiments on animals indicated cancerogenous effect of agarityn found in numerous species of the genus agaricus. derivatives of hydrazine are also carcinogens, resulting in colon tumors and other cancers in experimental animals. hydrazine derivatives were found in some species of the genus agaricus, and shiitake mushrooms (sadowska et al. 2004). these species should be evaluated with caution. škubla (2005) rightly says that no mushroom can be recommended for consumption. in the recent years, studies on the diversity of polysaccharides and their biological activity have been carried out at a fairly large scale. fan et al. (2006) indicate that more than 650 species of fungi representing more than 180 genera are known for containing polysaccharides that stimulate the immune system and act as anti-cancer agents. 94 k. kopczyński vitamins and minerals. in addition to selenium and β-glucans, fungi contain other substances valuable for human health. vitamin d should be indicated here in particular. a component of the cell walls of fungi – ergosterol – is converted into vitamin d2 in the presence of sunlight or other ultraviolet light source (marley 2009; babal 2011). this vitamin has an essential role in prevention of cancer, as it results in increased phagocytosis (absorption and destruction of cancer cells) and facilitating other immunomodulatory functions. it is also responsible for the absorption of calcium and phosphorus, which enables normal growth of bones and prevents osteoporosis (marley 2009). in laboratory studies, it has been found that vitamin d slows the growth of cancer cells and its administration in animal studies resulted in the decrease of the incidence of breast cancer by half (carper1995). tomasik (2008) lists the mushrooms as one of the four main sources of vitamin d in food. gertig (2007) indicates that significant amounts of vitamin d are contained in fish and mushrooms. for example, the edible parts of halibut contain about 4 μg, while porcini mushrooms (boletus) about 7 μg per 100 g of the product. in the recent exciting discoveries it has been found that mushrooms exposed to ultraviolet rays, either before or after the collection, quickly convert ergosterol to vitamin d2 in astonishing quantities. cases where vitamin d levels have increased hundredfold during the drying of mushrooms in the sun in comparison with the mushrooms dried in the dark are cited. this increase was so significant that the possibility of an overdose of vitamin d in people who had eaten mushrooms dried in the sun was considered, but toxicity of vitamin d assimilated from fungi was not recorded (marley 2009). another important influence of vitamin d is worth mentioning. in the course of cancer, formation of microvessels within the tumor (angiogenesis) plays an important role. the progression of cancer may be limited by limiting the functions of proangiogenic factors. one of the factors blocking the formation of new blood vessels needed for tumor growth is adequate diet (mraz et al. 2010). the use of this dietary vitamin d may be important, because vitamin d analogues are on the list of factors responsible for the inhibition of angiogenesis (obrocka et al. 2002; wcisło 2010). the importance of minerals is also stressed. providing large amounts of calcium from food significantly reduces the risk of developing colorectal cancer (hasik 2000). as already mentioned, vitamin d is responsible for the absorption of calcium, so it is involved in the cancer prevention. besides vitamin d, mushrooms contain, among others, vitamin b, c, e and k (marley 2009). particular importance of vitamin c and e in cancer prevention is indicated i.a. by hasik (2000) and unger and hildenbrand (2008). hasik (2000) states that nitrosoamines belong to the strongest carcinogens, while vitamin c has significant inhibitory effect on the formation of nitrosoamines (ber, gazella 2002). conclusions for 25 years klimuszko (1988) advocated the use of fungi in medicine and applied a delicate mushroom cure with good results in almost all cases for those whose organisms were ultimately exhausted due to various diseases. he concluded the immunomodulating properties 95 beneficial effect of edible mushrooms from the fact that they are often found next to the highly poisonous species, thus, for the balance of biological effects that has to be sustained in nature, they must have significant positive effects, even if imperceptible, but equal in strength to the poison. in the light of the above-described immunomodulatory and anti-cancerogenous properties of fungi, therapeutic effects of klimuszko’s fungal diets appear to be understandable. while consuming mushrooms and their immunomodulatory and anti-cancer components, the organism becomes more resistant, thus protected from disease; 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(in:) c. unger, j. weis (eds). onkologia. wydawnictwo medpharm polska, wrocław: 94–181. sadowska a., obidoska g., ruszkowska j., rumowska m., łata b. 2004. rakotwórcze i trujące substancje roślinne. wydawnictwo sggw, warszawa. škubla p. 2005. atlas grzybów. vydavatiel’stvo slovart – wydawnictwo solis, bratislava-warszawa. strach m. 2011. układ odpornościowy u ludzi starszych. infekcje i ich profi laktyka. materiały dydaktycz-układ odpornościowy u ludzi starszych. infekcje i ich profilaktyka. materiały dydaktyczne medycznego centrum kształcenia podyplomowego uniwersytetu jagiellońskiego. uj, kraków. szponar l., respondek w. 2000. choroby pierwotne na tle niedoborów żywieniowych. (in:) j. hasik, j. gawęcki (eds). żywienie człowieka zdrowego i chorego. pwn, warszawa: 95–110. tomasik p. 2008. witamina d. (in:) p. p. lewicki (ed.). leksykon nauki o żywności i żywieniu człowieka oraz polsko-angielski słownik terminów: 483. wydawnictwo sggw, warszawa. unger c., hildenbrand b. 2008. system immunologiczny i ochrona immunologiczna przed nowotworem. (in:) c. unger, j. weis (eds). onkologia. wydawnictwo medpharm polska, wrocław: 76–93. wcisło g. 2010. związki chemiczne w trakcie badań jako potencjalne leki i eksperymentalne sposoby terapii raka nerki. (in:) c. szczylik, g. wcisło (eds). rak nerki. termedia wydawnictwo medyczne, poznań: 297–322. żurowska k. 2010. grzyby, rośliny, beta-glukany – tajemnica poprawy odporności. medycyna estetyczna i anti-aging 1: 40–42. immunomodulacyjne i przeciwnowotworowe właściwości grzybów streszczenie niniejszy artykuł jest próbą syntezy wyników badań nad immunomodulacyjnymi i przeciwnowotworowymi właściwościami grzybów. dotychczas prezentowano zwykle naukowe doniesienia o którymś ze zdrowotnych składników zawartych w grzybach. dla kształtowania odporności organizmu i zapobiegania nowotworom duże znaczenie mają β-glukany i selen, a także witamina d, c i e. substancje te występują równocześnie w grzybach, które wskutek tego uzyskują szczególną wartość zdrowotną. zdaniem niektórych badaczy, stosowanie mieszanki różnych gatunków grzybów z szeroką różnorodnością glukanów może spowodować zwiększone działanie odpornościowe. glukany bowiem występują w niewiarygodnie różnorodnych konfiguracjach, odzwierciedlających zakres ich biologicznego działania. w ostatnich fascynujących odkryciach stwierdzono także, iż grzyby suszone na słońcu (wystawione na działanie promieni ultrafioletowych) szybko przetwarzają ergosterol – składnik ścian komórkowych grzybów – w witaminę d2 w zdumiewającej ilości. mitem okazuje się – w świetle współczesnych badań – rozpowszechnione mniemanie o jedynie smakowych i aromatycznych walorach grzybów. 2014-01-02t12:04:46+0100 polish botanical society introduced tuber aestivum spreading spontaneously in israel varda kagan-zur1*, tidhar turgeman2, yaron sitrit2, ofer danai3, yoram luzzati4, amnon bustan2, nurit roth-bejerano1 and segula masaphy5 1department of life sciences, and 2the jacob blaustein institute for desert research ben-gurion university of the negev, beer-sheva il-84105 3edible mushrooms and 5applied mycology and microbiology departments, migal, p.o. box 831, kiryat shmona il-11016, and tel hai college, upper galilee il-12210 4kibbutz bar’am, m.p. merom hagalil il-13860,*corresponding author. zur@bgu.ac.il kagan-zur v., turgeman t., sitrit y., danai o., luzzati y., bustan a., roth-bejerano n., masaphy s.: introduced tuber aestivum spreading spontaneously in israel. acta mycol. 47 (2): 175–177, 2012. a tuber melanosporum plantation established in 1994/5 on kibbutz bar’am (in the upper galilee, israel, fig. 1) gradually lost its t. melanosporum mycorrhiza. when checked in 1998, only about 70% of the trees maintained their original mycorrhiza (kagan-zur et al. 2001) and was deteriorating constantly. a number of trees perished. several different tree species were initially introduced. these included both acta mycologica vol. 47 (2): 175–177 2012 introduced species: quercus ilex q.pubescens corylus avellana local species: q.boissieri q. penduculifora q. calliprinus q. cerris q. libani q. ithaburensis truffle grove fig. 1. the plantation (adapted from google earth). 176 v. kagan-zur et al. fig. 2. tuber aestivum vittad. on the top: a transverse cut of a fruit body found on 2009. on the bottom: spores. fig. 3. t. aestivum vittad. yields. amount collected in the years: 2010 – 2.2 kg, 2011 – 0.0 kg, 2012 – 1.7 kg. introduced tuber aestivum 177 european and local oak species, as well as hazelnuts. the different tree species were randomly planted within the grove. the plantation setting was necessary to facilitate our identification of the most suitable host species for producing the best yields under israeli conditions. it was later found that local oak species produced higher yields as compared to introduced oak species, chosen for their reputation of being good t. melanosporum symbionts. a single t. melanosporum fruit body was collected in 1999 (kagan-zur et al. 2000), but none later. in 1999, seedlings were introduced into the plantation to fill the gaps between trees. these included, inadvertently, t. aestivum inoculated plants. between 2000 and 2008 no truffles were found. however, in july 2009, two fruit bodies were collected and studied. their inner colors when cut and spore morphology observed under a light microscope indicated that these fruit bodies had a greater resemblance to t. aestivum than to t. melanosporum (fig.2). indeed, molecular analyses proved these to be t. aestivum (turgeman et al. 2012). thus, the intentionally introduced t. melanosporum mycorrhiza was replaced by that of another, unintentionally introduced mycorrhizal fungus, t. aestivum. in 2010 an organized manual search was performed (by digging systematically around 125 trees in the plot). a total of 2.2 kg of fruit bodies for a calculated yield of about 6 kg/acre was recovered (fig. 3). this was comparable to yields reported for young commercial orchards elsewhere (southern woods nursery 2011). no fruit bodies were collected during 2011. however, in 2012 truffles were found both at the original plantation and, accidentally, at a grove within a research farm about 3.5 km away. the latter seem to be of the same origin as the originally introduced t. aestivum. in conclusion, the environmental conditions on the upper galilee, though unsuitable for t. melanosporum, fit the requirements of the more robust t. aestivum fungus, which thrives on local oak species and even spreads further. references kagan zur v., freeman s., luzzati y., roth-bejerano n., shabi e. 2000. emergence of the first black périgord truffle (tuber melanosporum) in israel. mycol veget. mediter. 15: 187–192. kagan-zur v., freeman s., luzzati y., roth-bejerano n., shabi e. 2001. survival of introduced tuber melanosporum mycorrhizas at two sites in israel as measured by its occurrence on mycorrhizas. plant & soil 229: 159–166. southern woods nursery: truffles. 2011. http://www.southernwoods.co.nz/documents/truffles.pdf turgeman t., sitrit y., danai o., luzzati y., bustan a., roth-bejerano n., kagan-zur v., masaphy s. 2012. introduced tuber aestivum replacing introduced tuber melanosporum: a case study. agroforestry systems 84: 337–343. 2014-01-02t12:13:19+0100 polish botanical society the lichen family parmeliaceae in poland. i. the genus parmotrema agnieszka jabłońska, magdalena oset and martin kukwa department of plant taxonomy and nature protection university of gdańsk, al. legionów 9, pl-80-441 gdańsk agnieszkajablonska82@wp.pl jabłońska a., oset m., kukwa m.: the lichen family parmeliaceae in poland. i. the genus parmotrema. acta mycol. 44 (2): 211–222, 2009. the paper presents the results of study on four parmotrema species in poland, p. arnoldii, p. crinitum, p. perlatum and p. stuppeum. they are mainly known from southern part of the country, mostly in the carpathians. p. perlatum is reported also from northern poland and central poland. for p. stuppeum, so far known from single locality only, three new sites are reported. all the species seem to be rare and endangered in poland. this work is the first part of a larger series which will present data on selected genera of the lichen family parmeliaceae in poland. key words: parmotrema, parmeliaceae, chemotaxonomy, foliose lichens introduction the lichen family parmeliaceae zenker (lecanorales, lecanoromycetes, ascomycota; see lumbsch, huhndorf 2007) comprises several genera of mostly foliose, but also few fruticose and crustose lichens (e.g., elix 1993; lumbsch, huhndorf 2007), and recently even the lichenicolous genus phacopsis tul. was proved to belong to the family (peršoh, rambold 2002). parmeliaceae is morphologically and chemically very diverse group of lichens, but generally it is characterized morphologically by a type of apothecial ontogeny and the presence of an ascomatal structure called a cupulate exciple (blanco et al. 2006 and literature cited therein). the family comprises about 1500 species and is thought to be one of the richest in species within lichenized ascomycota (blanco et al. 2006). it is also one of the most widely discussed group of lichens in terms of generic delimitation. for a long time only few genera were accepted, but extensive research, mostly by m. e. hale jr. (cf. hale 1974a, b, 1975, 1976a, b, c, 1986), led to segregation of morphologically and chemically more homogenic groups. however, those taxonomic concepts were acta mycologica vol. 44 (2): 211–222 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 212 a. jabłońska et al. not always followed and all segregates treated as doubtful (e.g., purvis et. al. 1992). recently, the taxonomy of parmeliaceae has been studied with molecular tools, and the importance of chemical, morphological and anatomical diagnostic characters have been re-evaluated (e.g., blanco et al. 2004a, b, 2005; divakar et al. 2006). numerous hale’s genera and those distinguished later by other lichenologists are still accepted, however, some of them appeared to be synonyms (blanco et al. 2004a, b, 2005; divakar et al. 2006). parmotrema a. massal. is one of the genera accepted at present, however in a wider sense then previously, as it also includes canomaculina elix & hale, concamerella w. l. culb. & c. f. culb., parmelaria awasthi and rimelia hale & fletcher, which were found to be phylogenetically nested within parmotrema s.str. (blanco et al. 2005). the genus is characterized by usually large foliose thalli with ciliate or eciliate margin, lack of pseudocyphellae, upper cortex consisting of a palisade plectenchyma (or rarely paraplectenchyma), pored or fenestrated epicortex, laminal, perforate or eperforate apothecia and usually simple rhizines (blanco et al. 2005). as many parmotrema species are very similar in morphology, the secondary chemistry plays an important role in the identification and taxonomy of these lichens. the substances found in this genus comprises atranorin and rarely usnic acid, which are present in the cortex, and several medullary compounds belonging to orcinol depsides, orcinol depsidones, ß-orcinol depsides, xanthones, aliphatic acids, pulvinic acid and derivatives and antraquinones (cf. hale 1965; blanco et al. 2005). for the determination of them colour reactions with simple chemical reagents (c, k and pd) were used in poland, however this method is not very reliable, as it can not distinguish lichen substances with the same type of reaction. for that reason specimens should be studied by thin-layer chromatography (tlc) . until now 4 taxa have been reported in poland (fałtynowicz 2003), and almost all of them have not been confirmed by tlc. some species were consider to be rather common, e.g., p. perlatum (huds.) m. choisy, but some were very rarely reported from few stands only, e.g., p. stuppeum (taylor) hale. our unpublished results show that frequency for some taxa is different. the aims of this paper, the first dealing with the revision of selected genera of the family parmeliaceae in poland, are to present the result of studies on the morphology, chemistry, distribution and habitat requirements of the genus parmotrema in the country. material and methods the present study is based on collection which are deposited in polish lichen herbaria (kram, krap, ktc, lbl, ugda). the morphology of the specimens were examined under the stereo microscope for thallus colour, shape of lobes, presence and abundance of cilia, and the type of soralia and isidia. the lichen substances were investigated by thin layer chromatography (tlc) following the methods described by orange et al. (2001). the chromatograms were developed in solvent c. the family parmeliaceae 213 all examined localities are mapped according to the atpol grid square system (zając 1978; modified by cieśliński, fałtynowicz 1993; see also kukwa et al. 2002). results parmotrema arnoldii (du rietz) hale, phytologia 28: 335 (1974). syn. parmelia arnoldii du rietz, nyt mag. naturvidensk. 62: 80 (1924) . characteristic of the species. parmotrema arnoldii is distinguished by the sublaminal soredia developing mostly at laciniae, narrow, mostly black non-rhizinate marginal zone, usually sparsely ciliate thallus margin and the presence of atranorin, alectoronic and α-collatolic acids, sometimes accompanied also by skyrin (= rhodophyscin). cortex reacts k+ yellow and medulla is kc+ pink-red, uv+ ice-blue, c– and pd– (hale 1965; louwhoff 2009a). in polish specimens all substances, except skyrin, were found. affinities. the species is distinguished from morphologically similar taxa producing soredia by the production of alectoronic and α-collatolic acids. there is no other species of parmotrema with these substances in europe (hale 1965), however this substances are produced in the morphologically somewhat similar cetrelia chicitae (w. l. culb.) w. l. culb. & c. f. culb. this species differs in the presence of pseudocyphellae and eciliate thallus margin (obermayer, mayrhofer 2007). habitat requirements. according to louwhoff (2009a) p. arnoldii usually grows amongst epiphytic mosses, especially on horizontal branches of trees and old shrubs fig. 1. distribution of parmotrema arnoldii in poland in the atpol grid square system. 214 a. jabłońska et al. in ± well-lit, mild and humid, undisturbed woodlands. in poland this lichen was collected on acer spp., alnus spp., fagus sylvatica, fraxinus excelsior and pyrus sp. in beech and black alder forests or in open places. distribution. in poland p. arnoldii is a rare mountain species, known only from the carpathians (fig. 1). most of the analyzed specimens originates from 1950–1960, and perhaps the species is close to extinction. parmotrema arnoldii is a temperate species of both, northern and southern hemispheres. this taxon occurs in europe, e.g., austria (hafellner, türk 2001), germany (hale 1965; scholtz 2000), great britain (coppins 2002), france (hale 1965), ireland (fox 2004), scandinavia (hale 1965; santesson et al. 2004), slovenia (suppan et al. 2000), spain (llimona, hladun 2001), switzerland (scheidegger et al. 2002) and ukraine (kondratyuk et al. 1998). outside europe it was reported from macaronesia, africa, asia, and north and south americas (e.g., hale 1965; hafellner 1995; louwhoff 2009a; pišút 2009). specimens examinated. poland. fd 95 – beskid mały mts., gładkie pass, above przełęcz kocierska pass, alt. ca 650 m, on acer platanoides, 10 apr. 1962, leg. j. nowak (kram-l 9466); the same locality, alt. ca 700 m, on fraxinus excelsior, 4 oct. 1962, leg. j. nowak (kram-l 10004). gd 59 – tatry mts., dolina białego valley, on acer pseudoplatanus, 10 aug. 1925, leg. j. motyka (kram-l 10150). ge 21 – gorce mts., near kamienica village, in the wojtasy forest, alt. 660 m, alder forest, on alnus sp., 20 aug. 1967, leg. k. glanc (kram-l 35413). gf 59 – bieszczady zachodnie mts., pszczeliny village, alt. 700 m, alder forest, on alnus sp., 25 sept. 1958, leg. k. glanc (kram-l 35420, in the specimen of parmotrema perlatum). gf 68 – bieszczady zachodnie mts., bieszczadzki national park, beskidnik mt., alt. 740 m, beech forest, on fagus sylvatica, 28 aug. 1957, leg. k. glanc (kram-l 35425); by the beskidnik stream, beech forest, on fagus sylvatica, 1957, leg. k. glanc (kram-l 35428). gf 69 – bieszczady zachodnie mts., bieszczadzki national park, caryńskie village, near przełęcz przysłup pass, alt. 700 m, on pyrus sp., sept. 2005, leg. r. kościelniak (krap). parmotrema crinitum (ach.) m. choisy, bull. mens. soc. linn. soc. bot. lyon 21: 175 (1952). syn. parmelia crinita ach., syn. lich.: 196 (1814). characteristic of the species. parmotrema crinitum is characterized by the presence of marginal and laminal isidia, which are often ciliate, lower surface black with a broad brown erhizinate marginal zone and the production of atranorin (cortex) and stictic acid complex, including menegazziaic acid (medulla). isidia wary from sparse, simple, cylindrical or lobulate to coralloid-branched and forming dense clusters. cortex reacts k+ yellow, whereas medulla is c–, k+ yellow, kc+ yellow, pd+ yellow-orange (hale 1965; louwhoff 2009a). the chemistry agrees well with that reported by hale (1965) and louwhoff (2009a). affinities. the species is easily recognizable by the presence of isidia, broad bare marginal zone and the production of stictic acid complex. parmelinopsis horrescens (taylor) elix & hale also has ciliate-isidia, but it is distinguished by the smaller thallus, the lower surface that is rhizinate to the margins and the lack of stictic acid complex (louwhoff 2009b). so far this species has not been found in poland. marginal and laminal isidia of similar shape to those present in parmotrema crinitum are present also in platismatia glauca (l.) w. l. culb. & c. f. culb., but that species lacks cilia, does not react with pd (stictic acid complex absent) and produces caperatic acid (culberson, culberson 1968). habitat requirements: the species usually grows on bark of deciduous trees in old forests or in open situations (fałtynowicz 2003; louwhoff 2009a). in poland it the family parmeliaceae 215 was so far reported only as epiphyte on abies alba, acer pseudoplatanus, alnus spp., fagus sylvatica (ca 58% of polish records) and picea abies, usually in beech forests or along streams. distribution. according to fabiszewski (1968) and fałtynowicz (2003 and literature cited therein) p. crinitum was reported from the carpathians and sudety mts., however this study confirm its occurrence in the carpathians (material from sudety mts. not traced). known distribution of the species is presented on figure 2. this lichen is known only from old collections, and perhaps it is already extinct in the territory of poland. p. crinitum is a cosmopolitan species, widespread throughout tropical and temperate areas and even subboreal forests (hale 1965; louwhoff 2009a). it was reported in europe from, e.g., austria (hafellner, türk 2001), belgium (diederich et al. 2009), the czech republic (liška et al. 2008), france and hungary (hale 1965), germany (scholz 2000), great britain (hale 1965; coppins 2002), italy (hale 1965), ireland (fox 2004), luxembourg (diederich et al. 2009), portugal (hale 1965), scandinavia (hale 1965; santesson et al. 2004), slovakia (lisická 2005), slovenia (sup-suppan et al. 2000), spain (hale 1965) and ukraine (kondratyuk et al. 1998). outside europe it is known from africa, asia, australia, macaronesia, and north and south america (e.g., hale 1965; hafellner 1995; louwhoff and elix 1999; louwhoff 2009a; mccarthy 2009). specimens examinated. poland. gd 59 – tatry zachodnie mts., łysanki mt., on picea abies, 29 aug. 1925, leg. j. motyka (kram-l 10138). gf 47 – bieszczady zachodnie mts., by solinka river, beech forest, on abies alba, 13 aug. 1956, leg. k. glanc (kram-l 36990). gf 57 – bieszczady zachodnie mts., falowa mt., se of cisna village, on alnus sp., 17 june 1954, leg. t. sulma (ugda-l 2561); krywe village, by dołżycki stream, near road to dołżyca village, alt. 600 m, on alnus sp., 28 june 1958, leg. k. glanc (kram-l 36987). gf 59 – bieszczady zachodnie mts., stuposiany village, alt. 640 m, beech forest, on fig. 2. distribution of parmotrema crinitum in poland in the atpol grid square system. 216 a. jabłońska et al. fagus sylvatica, 23 june 1957, leg. k. glanc (kram-l 36988); se of pszczeliny village, alt. 690 m, by the stream, on abies alba, 29 june 1959, leg. k. glanc (kram-l 38926); w of pszczeliny village, alt. 700 m, alder forest, on alnus sp., 24 sept. 1958, leg. k. glanc (kram-l 36993). gf 68 – bieszczady zachodnie mts., bieszczadzki national park, by beskidnik stream, alt. 740 m, beech forest, on acer pseudoplatanus, 1 sept. 1962, leg. k. glanc (ugda-l 12243); beskidnik mt., alt. 740 m, beech forest, on fagus sylvatica, 28 aug. 1957, leg. k. glanc (kram-l 36991); near wetlina village, by wielki bukowy stream, ca alt. 750 m, beech forest, on fagus sylvatica, 18 aug. 1958, leg. z. tobolewski (ugda-l 4476); by wielki lutowy stream, alt. ca 750 m, beech forest, on fagus sylvatica, 18 aug. 1958, leg. k. glanc (kram-l 36994, 36996, 35997, 36999 & 37000); s slope of dział mt., beech forest, on fagus sylvatica, 19 aug. 1958, leg. k. glanc (kram-l 35998 & 36989). gg 60 – bieszczady zachodnie mts., bieszczadzki national park, s slope of tarnica mt., alt. 900 m, beech forest, on fagus sylvatica, 22 june 1956, leg. k. glanc (kram-l 36992). poorly localized specimen. tatra mts., 1889, leg. w. boberski (kram-l 20176). parmotrema perlatum (huds.) m. choisy., bull. mens. soc. linn. soc. bot. lyon 21: 174 (1952). syn. lichen perlatus huds., fl. angl.: 448 (1762), parmelia perlata (huds.) ach., meth. lich.: 214 (1803), parmotrema chinense auct. non (osbeck) hale & ahti. characteristic of the species. the diagnostic characters of p. perlatum are the conspicuous submarginal soralia, revolute lobes, narrow, shiny and often rugose naked zone of lower side along the margin. the species contains atranorin, stictic acid complex, including stictic, constictic and menegazziaic acids with other related substances. cortex reacts k+ yellow, whereas medulla is c−, k+ yellow and pd+ orange(louwhoff 2009a). the chemistry of polish agrees with the previously reported results. affinities. parmotrema perlatum can be separated from other sorediate and marginally ciliate species, p. arnoldii (du rietz) hale and p. robustum (degel.) hale, by the presence of stictic acid complex (hale 1965; louwhoff 2009a). in the spot test fig. 3. distribution of parmotrema perlatum in poland in the atpol grid square system. the family parmeliaceae 217 reactions, p. perlatum is similar to p. reticulatum (taylor) m. choisy, but the latter is readily distinguished by the network of hair-line cracks on the upper surface, the soredia at tips of incised lobe margins and the presence of salazanic acid (louwhoff 2009a); so far it has not been reported from poland. few specimens of p. perlatum were found under cetrelia cetrarioides (delise & duby) w. l. culb. & c. f. culb. s.l. this taxon, and also other cetrelia species also develop wide lobed thalli, but they differ in the presence of pseudocyphellae, eciliate thallus margin and they never produce stictic acid complex (culberson, culberson 1968; obermayer, mayrhofer 2007). also platismatia glauca is morphologically similar to parmotrema perlatum, but it lacks marginal cilia, has isidia and a different chemistry (caperatic acid) (culberson, culberson 1968). habitat requirements. parmotrema perlatum is sensitive to air pollution and occurs on well-lit, neutral to somewhat acid-barked, broad-leaved trees, also frequently on siliceous rocks and walls, coastal rocks, usually where illumination is moderate to good (lowhoff 2009a). in poland this lichen was found mostly on bark of alnus spp. and fagus sylvatica, but single specimens were also collected on bark of fraxinus excelsior and picea abies. distribution. in poland p. perlatum was reported several times from southern poland (fałtynowicz 2003). after the revision of all available specimens most of those records appeared to be correct. some additional localities are added here, with records from northern and central poland (the locality from central poland also included in paper by łubek 2009). the known distribution of the species is presented on figure 3. as all examined specimens were collected before 1960, the species is perhaps close to extinction. the species is a cosmopolitan lichen widespread throughout temperate and tropical areas (hale 1965; louwhoff 2009a), known from many european countries, e.g., austria (hafellner, türk 2001), belgium (diederich et al. 2009), the czech republic (liška et al. 2008), france (hale 1965), germany (scholtz 2000), great britain (coppins 2002), ireland (hale 1965), italy (hale 1965), luxembourg (diederich et al. 2009), the netherlands (hale 1965; aptroot et al. 2004), portugal (hale 1965), scandinavia (santesson et al. 2004), slovakia (lisická 2005), spain (hale 1965; llimona, hladun 2001) and ukraine (kondratyuk et al. 1998). the species has been also recorded from macaronesia, africa, asia, australia, north america and south america (e.g. hale 1965; hafellner 1995; louwhoff, elix 1999; mccarthy 2009; pišút 2009). specimens examinated. poland. ad 96 – wysoczyzna elbląska high plain, brzozowo forest district, ne of milejewo village, old beech forest, on fagus sylvatica, 20 july 1957, leg. t. sulma (ugda-l 15000). ee 60 – wyżyna przedborska upland, niecka włoszczowska basin, near oleszno nature reserve, forest section no. 73, on fraxinus excelsior, 11 march 2008, leg. a. łubek (ktc). eg 91 – roztocze środkowe, zwierzyniec village, by świerszcz river, on alnus glutinosa, 1969, leg. j. bystrek (lbl). gf 57 – bieszczady zachodnie mts., w slope of krzemienna mt., alt 650 m, on picea abies, 28 july 1958, leg. k. glanc (kram-l 37051); krywe, by the road to dołżyca village, by dołżycki stream, ca 580 m, on alnus glutinosa, 28 july 1958, leg. k. glanc (kram-l 37050, in specimen of cetrelia cetrarioides s.l.); near krywe village, by the turist path to cisna village, alt. 550 m, alder forest, by the stream, on alnus sp., 30 july 1958, leg. k. glanc (kram-l 35416); cisna village, by the road to krywe village, alt. 550 m, alder forest, by the stream, on alnus sp., 30 july 1958, leg. k. glanc (kram-l 35418); se of cisna village, slopes of falowa mt., in deep gorge, on alnus sp. and bark of unidentified tree, 17 july 1954, leg. t. sulma (ugda-l 2511 & 2528). gf 59 – bieszczady zachodnie mts., pszczeliny village, s of stuposiany village, alt. ca 650 m, mixed forest by the stream, on alnus sp., 25 sept. 1958, leg. z. tobolewski (ugda-l 4477); pszczeliny 218 a. jabłońska et al. village, alt. 700 m, alder forest, on alnus sp., 25 sept. 1958, leg. k. glanc (kram-l 35420); the same locality, 24 sept. 1958, leg. k. glanc (ugda-l 35422). gf 68 – bieszczady zachodnie mts., bieszczadzki national park, by beskidnik stream, on fagus sylvatica, s.dat., leg. k. glanc (kram-l 35429); by wielki lutowy stream, alt. ca 750 m, beech forest, on fagus sylvatica, 18 aug. 1959, leg. k. glanc (kram-l 36994, 36996 & 35997, all admixed in specimens of parmotrema crinitum). gf 69 – bieszczady zachodnie mts., bieszczadzki national park, bereżki village, by the wołosaty stream, alt. 600 m, on alnus sp., 21 sept. 1958, leg. k. glanc (kram-l 37055); the same locality, 22 sept. 1958, leg. k. glanc (kram-l 35424); near road to wołosatka stream, alt. 840 m, on fagus sylvatica, 22 june 1956, leg. z. tobolewski (kram-l 35419); by the rzeczyca stream, by the road between villages ustrzyki górne and brzegi (berehy) górne, alt. 680 m, alder forest, on alnus sp., 27 aug. 1957, leg. k. glanc (kram-l 35423); the same locality, 7 july 1959, leg. k. glanc (kram-l 35415); se of pszczeliny village, alt. 690 m, by the stream, on alnus sp., 29 july 1959, leg. k. glanc (kram-l 35414). gg 70 – bieszczady zachodnie mts., bieszczadzki national park, by the wołosatka stream, near tarnica mt., alt. 840 m, on fagus sylvatica, 21 june 1956, leg. k. glanc (kram-l 35417). parmotrema stuppeum (taylor) hale, phytologia 28: 339 (1974). syn. parmelia stuppea taylor, london j. bot. 6: 175 (1847); for more synonyms see hale (1965). characteristic of the species. parmotrema stuppeum is characterized by thallus loosely attached to the bark, black and sparsely rhizinate lower surface, continuous (not finely reticulately cracked) upper cortex, soralia formed on the top of laciniae and the presence of atranorin in cortex and salazinic acid in medulla (hale 1965; sipman 2005). cortex reacts k+ yellow, whereas medulla is c and kc–, k+ yellow turning red, and pd+ orange-red. the chemistry of polish specimens agrees with previously reported information. affinities. the species can be separated from other sorediate parmotrema species with continuous cortex by the presence of salazinic acid. this substance is produced by p. reticulatum (tayl.) m. choisy [syn. rimelia reticulata (tayl.) hale & fletcher] and p. subreticulatum (tav.) hale, but these taxa has finely, but distinctly reticulately cracked and maculate upper cortex (hale 1965; sipman 2005; louwhoff 2009a); so far they have not been reported from poland. in poland most specimens of p. stuppeum were determined as p. chinense, a synonym of p. perlatum (see above). both taxa have pd+ orange to orange-red medulla, but it is due to salazinic acid in p. stuppeum, whereas in p. perlatum the reaction is caused by stictic acid and related substances. they can also be separated without tlc; medulla of p. perlatum is k+ yellow, but in p. stuppeum the yellow colour turns red (hale 1965; sipman 2005; louwhoff 2009a). habitat requirements. in poland p. stuppeum is a corticolous lichen. it was collected almost exclusively on fagus sylvatica, but once it was found on bark of pyrus communis. distribution. in poland the species has been so far reported only once by sulma and fałtynowicz (1988). here we report three new localities, two from bieszczady mts. and one from roztocze. the known distribution of the species is presented on figure 4. it appears as a rare and an endangered lichen in poland, as only one contemporary record is known. according to hale (1965) p. stuppeum is common in the mountain of north america, central america and europe; in africa and asia it is more rare. in europe the species has been reported from austria (hafellner, türk 2001), the czech republic (liška et al. 2008), germany (hale 1965; scholtz 2000), portugal (hale 1965), spain (llimona, hladun 2001), switzerland (scheidegger et al. 2002) and the family parmeliaceae 219 ukraine (kondratyuk et al. 1998). it was also reported from the netherlands, but it has been recently considered as misidentification; the records belong to p. subreticulatum (aptroot et al. 2008). specimens examinated. poland. eg 93 – roztocze środkowe, majdan ruszowski village, 50°37’n, 23°18’e, on fagus sylvatica, 21 aug. 1933, leg. t. sulma (ugda-l 2555). gf 57 – bieszczady zachodnie mts., s slope of łopiennik mt., alt. ca 950 m, on fagus sylvatica, 30 june 1958, leg. k. glanc (kram-l 35421). gf 58 – bieszczady zachodnie mts., bieszczadzki park narodowy, suche rzeki, alt. 670 m, on pyrus communis, aug. 1995, leg. r. kościelniak (krap). gf 68 – bieszczady zachodnie mts., by beskidnik stream, beech forest, on fagus sylvatica, 1957, leg. k. glanc (kram-l 35426, ugda-l 3547, also in specimen of parmotrema arnoldii, kram-l 35428); the same locality, on fagus sylvatica, s.dat., leg. k. glanc (kram-l 35429, as a mixture in specimen of p. perlatum). conclusions four species of parmotrema are known to occur in poland, p. arnoldii, p. crinitum, p. perlatum and p. stuppeum. they have been mainly found in southern part of the country, mostly in the carpathians. only p. perlatum is known from two localities in northern and central poland. the study has shown, that the secondary lichen metabolites are very useful character in determination of species; the analyzes of lichen secondary metabolites led to discovery of 3 new localities of p. stuppeum. so far the species has been reported only from one locality. fig. 4. distribution of parmotrema stuppeum in poland in the atpol grid square system. 220 a. jabłońska et al. at present, only p. perlatum and p. stuppeum are known to be represented by recent collections. it appeared, that p. arnoldii and p. crinitum have not been collected since 1962. thus, all parmotrema species should be considered as in great danger of extinction in poland. possibly two of them, p. arnoldii and p. crinitum, are already extinct in poland. acknowledgements. we are grateful to the curators of herbaria for the loan of specimens and the reviewer for helpful comments. this paper is dedicated to professor krystyna czyżewska (łódź) on the occasion of her anniversary. reference aptroot a., herk, van c. m., sparrius l. b., spier j. l. 2004. checklist van de nederlandse korstmossen en korstmosparasieten. buxbaumiella 69: 17–55. aptroot a., spier j. l., jordaens d. 2008. parmotrema pseudoreticulatum, de verbeterde determinatie van parmotrema stuppeum (gewimperd schildmos). buxbaumiella 80: 9 –12. blanco o., crespo a., divakar p. k., elix j. a., lumbsch h. t. 2005. molecular phylogeny of parmotremoid lichens (ascomycotina, parmeliaceae). mycologia 97: 150–159. blanco o., crespo a., elix j. a., hawksworth d. l., lumbsch h. t. 2004a. a molecular phylogeny and a new classification of parmelioid lichens containing xanthoparmelia-type lichenan (ascomycota: lecanorales). taxon 53 (4): 959–975. blanco o., crespo a., divakar p. k., esslinger t. l., hawksworth d. l., lumbsch h. t. 2004b. melanelixia and melanohalea, two new genera segregated from melanelia (parmeliaceae) based on molecular and morphological data. mycol. res. 108 (8): 873–884. blanco o., crespo a., ree r. h., lumbsch h. t. 2006. major clades of parmelioid lichens (parmeliaceae, ascomycota) and the evolution of their morphological and chemical diversity. mol. phyl. evol. 39: 52–69. cieśliński s., fałtynowicz w. 1993. note from editors. 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(eds). 1992. the lichen flora of great britain and ireland. natural history museum publications & british lichen society, london, 710 pp. santesson r., moberg r., nordin a., tønsberg t., vitikainen o. 2004. lichen-forming and lichenicolous fungi of fennoscandia. museum of evolution, uppsala university, uppsala, 359 pp. scheidegger c., clerc p., dietrich m., frei m., groner u., keller c., roth i., stofer s., vust m. 2002. rote liste der gefährdeten arten der schweiz: baumund erdbewohnende flechten. buwal-reihe vollzug umwelt, bundesamt für umwelt, wald und landschaft buwal, bern, und eidgenössische forschungsanstalt wsl, birmensdorf, conservatoire et jardin botaniques de la ville de genève cjbg, 124 pp. 222 a. jabłońska et al. scholz p. 2000. katalog der flechten und flechtenbewohnenden pilze deutschlands. schriftenreihe vegetationsk. 31: 1–298. sipman h. j. m. 2005. mason hale’s key to parmotrema, revised edition: key to wide-lobed parmelioid species occurring in tropical america (genera canomaculina, parmotrema, rimelia, rimeliella). http://www.anbg.gov.au/abrs/lichenlist/introduction.html [15.10.2009]. sulma t., fałtynowicz w. 1988. materiały do rozmieszczenia porostów z rodziny parmeliaceae w polsce. acta mycol. 23 (1): 107–123. suppan u., prügger j., mayrhofer h. 2000. catalogue of the lichenized and lichenicolous fungi of slovenia. biblioth. lichenol. 76: 1–215. zając a. 1978. atlas of distribution of vascular plants in poland (atpol). taxon 27: 481–484. porosty z rodziny parmeliaceae w polsce. i. rodzaj parmotrema streszczenie w polsce poznano dotychczas cztery gatunki z rodzaju parmotrema, p. arnoldii, p. crinitum, p. perlatum i p. stuppeum. gatunki te były publikowane bez analizy wtórnych metabolitów, co mogło skutkować nieprawidłowym oznaczeniem części okazów. obecnie zrewidowano cały dostępny materiał zielnikowy przy użyciu chromatografii cienkowarstwowej (tlc). w jej wyniku potwierdzono występowanie wszystkich czterech taksonów znanych dotychczas w kraju. w większości wypadków przedstawiciele tego rodzaju występują w polsce południowej, głównie w karpatach. p. perlatum został podany także z jednego stanowiska w polsce północnej (wysoczyzna elbląska) i jednego − w polsce środkowej (niecka włoszczowska). dla p. stuppeum podano trzy nowe stanowiska na terenie kraju; dotychczas porost ten znany był tylko z jednego stanowiska w bieszczadach. jak się wydaje, wszystkie potwierdzone gatunki są silnie zagrożone wyginięciem w kraju, gdyż większość okazów zbierana była przed rokiem 1960. rozmieszczenie zrewidowanych taksonów przedstawiają ryciny 1–4. 2014-01-01t11:49:32+0100 polish botanical society the lichen genus pertusaria in poland i. p. multipuncta and p. ophthalmiza magdalena oset and martin kukwa department of plant taxonomy and nature protection university of gdańsk, legionów 9, pl-80-441 gdańsk magdalenasyrek@wp.pl, dokmak@univ.gda.pl oset m., kukwa m.: the lichen genus pertusaria in poland i. p. multipuncta and p. ophthalmiza. acta mycol. 45 (2): 231–238, 2010. the paper presents results of study on two morphologically very similar lichens in poland, p. multipuncta (turner) nyl. and p. ophthalmiza (nyl.) nyl. (pertusariales, ascomycota). previously, specimens were determined using only thallus characters and spot test reaction, with no data on lichen substances. this led to several misidentifications. after the revision of all available material of p. multipuncta from poland it appeared to be reported correctly only from one locality in gorce mts. most of other specimens belong to p. ophthalmiza, which has not been recorded in polish lichenological literature until 2008. key words: pertusariales, chemotaxonomy, neglected lichens, distribution introduction in poland 40 taxa of the genus pertusaria dc. have been reported so far (see hanko 1983; fałtynowicz 2003; kossowska 2008). some species were considered to be rather common (e.g., p. albescens (huds.) m. choisy & werner, p. amara (ach.) nyl.), but some were very rarely reported (e.g., p. melanochlora (dc.) nyl.). taxonomic status of few species is still very poorly known (e.g., p. inaequalis erichsen), and they need revision of type collections. up to now there has been no detailed study of the genus pertusaria in poland. many species are often sterile, and produce only soredia or isidia; these taxa are often superficially rather similar and cause difficulties in the identification. in such cases, lichen substances play a very important role in the determination of species. secondary chemistry is also very important in non sorediate, fertile taxa, which sometimes differ only in the chemical content (see dibben 1980; hanko 1983; archer 1997; chambers et al. 2009). however, so far only very few species have been confirmed by acta mycologica vol. 45 (2): 231–238 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 232 m. oset and m. kukwa thin layer chromatography (tlc) in the country. previously, specimens were determined using only thallus characters and spot test reaction, and very rarely the ascus type was studied. this could have led to many misidentifications. based on morphology and tlc-data, the present paper focuses on two taxa, namely p. multipuncta (turner) nyl. and p. ophthalmiza (nyl.) nyl., which are very similar in morphology, but have different chemistry. both produce ascomata, which are covered with soredia-like granules, and thus resemble soralia. p. multipuncta was reported in poland more often than p. ophthalmiza, as it could have been easily determined with the aid of the key by nowak and tobolewski (1975). until recently p. ophthalmiza has been reported from poland only by hanko (1983), who cited single, old collection. that record was not included in the recent polish checklist of lichens and lichenicolous fungi (see fałtynowicz 2003), and its presence in the country has remained unnoticed until the paper by kukwa et al. (2008). these authors reported also new polish localities of the species. as preliminary study suggested that the name p. multipuncta might have been misapplied in poland for the material of p. ophthalmiza, we decided to revise all available specimens of both taxa. the aim of this paper is to present the results of study on the chemistry, morphology, habitat requirements and distribution of p. multipuncta and p. ophthalmiza in poland. this paper is the first of a series of articles devoted to a revision of pertusaria in poland. material and methods the present study is based on the specimens deposited in the polish lichen herbaria: gpn, ktc, kram, lbl, lod and ugda. the morphology of the specimens was studied using a stereomicroscope. the thickness, morphology and colour of thallus, colour, shape and size of apothecia were noted. secondary metabolites were identified by thin layer chromatography (tlc) according to the methods of orange et al. (2001). the chromatograms were developed in solvent c. localities of all polish material examined are mapped according to the atpol grid square system (zając 1978; modified by cieśliński and fałtynowicz 1993; see also kukwa et al. 2002). characteristic of the genus pertusaria the genus pertusaria (pertusariales, ascomycota; see schmitt et al. 2006) includes corticolous, saxicolous, and terricolous crustose taxa, most of which do not grow on pure limestone or other strongly calcareous substrata (fałtynowicz 2003; chambers et al. 2009). many species occur in dry, open habitats, but some prefer humid localities (nowak, tobolewski 1975; fałtynowicz 2003; chambers et al. 2009). the genus is cosmopolitan and its members are found from the arctic and antarctic to the tropics in both hemispheres (e.g., dibben 1980; hanko 1983; archer 1997; chambers et al. the lichen genus pertusaria 233 2009). nevertheless, there are still substantial gaps in the distribution of many taxa in various regions of the world. in many cases, published data should be treated with caution, since the determination of species is often based mainly on morphological characters, but not secondary chemistry, which is an important systematic character set (e.g., dibben 1980; hanko 1983; archer 1997; chambers et al. 2009). the taxonomy of pertusaria and the delimitation from other phylogeneticaly related genera are still not satisfactorily settled. traditionally, members of the genus pertusaria are characterized by rather large (sometimes up to 4 mm in diameter) apothecia with hemiangiocarpous type of development, with open or almost closed, perithecia-like disk, hamathecium of branched and richly anastomosing, lax paraphysoids, thick-walled, amyloid asci with 1–8 spores, and thick spore wall. thallus is crustose, moderately thick or ± immersed, continuous to rimose-cracked, fissuredareolate or warted. in many species soralia and/or isidia are produced. in some taxa apothecial disk can be occluded with granular soredia and appearing soralium-like. secondary lichen metabolites are very diverse in pertusaria, and these can be divided into four chemosyndromes, each consisting of either xanthones, fatty acids, depsides or depsidones (e.g., dibben 1980; hanko 1983; archer 1997; chambers et al. 2009). after the recent molecular studies by schmitt and lumbsch (2004), pertusaria appeared as polyphyletic in its current circumscription and can be divided into three well supported groups, pertusaria s.str.-group, varicellaria-group and variolariagroup. the first one is characterized by amyloid, 2–8-spored asci with distinctive ocular chamber, non-amyloid hymenial gel, thick or thin walled spore wall, and the presence of chlorinated xanthones, gyrophoric and planaic acids. apothecia can be open with plane disc or closed, and then resembling perithecia. members of the varicellaria-group has strongly amyloid, 1–2-spored asci with no recognizable apex structures, strongly amyloid hymenial gel, ± thick spore wall, disciform apothecia, production of lecanoric acid and absence of chlorinated xanthones. at present 3 taxa of pertusaria are known to belong in this group, p. hemisphaerica (flörke) erichsen, p. lactea (l.) arnold and p. velata (turner) nyl., and they are close to the monotypic genus varicellaria nyl. (with v. rhodocarpa (körb.) th. fr.). the third, variolariagroup, is characterized by strongly amyloid, 1-spored asci with no recognizable apex structures, weakly or not amyloid hymenial gel, thin spore wall, open apothecia, production of picrolichenic and thamnolic acids, and absence of chlorinated xanthones (schmitt and lumbsch 2004). however, since that time no taxonomical segregation has been proposed yet, therefore all taxa are still kept in pertusaria. results and discussion pertusaria multipuncta (turner) nyl., lich. scand. (uppsala): 179 (1861). syn. variolaria multipuncta turner, trans. linn. soc. london 9: 137 (1806) characteristic of the species. p. multipuncta is distinguished from superficially similar species by the white, well-delimited, scattered, sorediate warts containing 234 m. oset and m. kukwa apothecia, 1-spored asci and the chemistry (physodalic acid and often protocetraric acid) (hanko 1983; chambers et al. 2009). thallus of the species can be very variable; on sheltered, smooth bark it is often thin, continuous and even, but in some exposed habitats it is more robust and the surface is coarsely warted (chambers et al. 2009). thallus and soralia of p. multipuncta are k+ yellow, c–, kc+ yellow, pd+ orange-red, uv– or faintly glaucous (chambers et al. 2009). in polish specimen physodalic and protocetraric acids were detected. affinities. in europe p. multipuncta can be mistaken for the morphologically similar p. ophthalmiza. both species produce similar soralium-like apothecia, but they can be easily separated by spot test reaction pd and thin layer chromatography: p. multipuncta reacts pd+ orange-red and contains physodalic acid, often together with protocetraric acid, and p. ophthalmiza is pd– and has fatty acids (hanko 1983; chambers et al. 2009). they differ also morphologically. p. multipuncta has fertile thallus warts measuring 0.5–1.5 mm diam., wide apothecial disc, and continuous and ± even exciple; in p. ophthalmiza the fertile warts are smaller (0.3–0.8 mm diam.), disc is up to twice narrower, and the exciple irregular and crenate (chambers et al. 2009). p. albescens and p. amara can also resemble p. multipuncta in morphology, but they are usually sterile, soredia are coarse (very often forming consoredia) and produced in true soralia; they posses also different lichen substances: p. albescens contains fatty acids, whereas p. amara picrolichenic acid, often accompanied by protocetraric acid (tønsberg 1992; chambers et al. 2009). habitat. p. multipuncta is a typical epiphytic lichen species, but it can rarely grow also on siliceous rocks (chambers et al. 2009). in poland it was found on bark of fagus sylvatica in beech forest. distribution. in poland p. multipuncta was reported from northern and southern part of poland, but rather scarcely (e.g., nowak and tobolewski 1975; fałtynowicz 2003 and literature cited therein; motiejūnaitė et al. 2004). after the revision of all available material from poland it appeared that most records were based on misidentifications. only one specimen from gorce mts appeared to belong to this species. most of other specimens belong to p. ophthalmiza, which has not been recorded in polish lichenological literature until 2008 (see kukwa et al. 2008, also below). some few additional specimens identified as p. multipuncta were rather small and difficult to identify, however they certainly do not belong either to p. multipuncta or p. ophthalmiza. p. multipuncta species was reported from many countries, however, as this name was often misapplied in the past (see hanko 1983), several records are doubtful. confirmed records of p. multipuncta appears to be confined to western and southern europe, where it has been so far reported from belgium, british isles, denmark, france, germany, italy, luxembourg, slovenia, spain, sweden (hanko 1983; suppan et al. 2000; søchting and alstrup 2002; nimis and martellos 2008; diederich et al. 2009). according to chambers et al. (2009) it occurs also in macaronesia, africa and asia. specimen examined. poland. ge 21 – gorce mts, gorce national park, area of wspólny potok stream, in kamienica stream valley, alt. 920 m, on fagus sylvatica, 9 june 1997, leg. p. czarnota (gpn 1583/94). specimen examined for comparison. norway. hordland, prov. asköy par., s of lake askevatnet, 60º29’n, 05º11’e, alt. 20 m., on branches of sorbus aucuparia, 11 june 1984, leg. t. tønsberg 884 & moberg, lich. sel. exs. upsal. 135 (kram-l 43334). the lichen genus pertusaria 235 pertusaria ophthalmiza (nyl.) nyl. – flora 48: 354 (1865). syn. pertusaria multipuncta var. ophthalmiza nyl., lich. scand.: 180 (1861), pertusaria multipuncta var. conferta erichsen, rabenh. kryptog.-flora von deutschland, österreich und der schweiz, part ix, 5(1): 610 (1936) [note: this variety was described from poland; see hanko (1983)] characteristic of the species. this species is anatomically and morphologically very similar to p. multipuncta, but it can be distinguished predominantly by the different chemical constituents; for the differences see under p. multipuncta. according to hanko (1983) and chambers et al. (2009), p. ophthalmiza contains 4 fatty acids, but in most polish specimens 2–3 fatty acids were found. only in three specimens 4 substances were found, but also 1 was detected in two samples. this variation is most probably caused by rather low quantity of material taken for the analyses, as most specimens were quite small. affinities. p. ophthalmiza can be mistaken for chemically and, when young, also morphology similar p. albescens. both species produce fatty acids, but they predominantly differ in thallus characters: p. albescens produces true soralia with granular soredia, whereas the sorediate warts of p. ophthalmiza contain apothecia (hanko 1983; tønsberg 1992; chambers et al. 2009). superficially p. ophthalmiza can resemble also ochrolechia turneri (sm.) hasselrot. this species often produces regular and separate soralia, which may resemble fertile warts of p. ophthalmiza. both taxa can be very easily separated by the c reaction of soralia and content of lichen substances. p. ophthalmiza is c negative and produces fatty acids, whereas soralia of o. turneri react c+ yellow and the species contains variolaric acid (hanko 1983; tønsberg 1992; kukwa 2008). habitat. research on polish material indicated that p. ophthalmiza is a corticolous species. it has been rather frequently found on tilia cordata (13 specimens). much less commonly it was found on alnus spp. (4 specimens), carpinus betulus (2 specimen) and fraxinus (1 specimen), always in old, humid woodland. according to chambers et al. (2009) this taxon can occur on acid bark (chiefly birches, oaks and coniferous trees) and overgrow epiphytic mosses, but in poland it appears to be strictly corticolous on deciduous trees with more neutral bark. distribution. p. ophthalmiza is much more widely distributed in europe than p. multipuncta, and it is known, e.g., from austria (hafellner and türk 2001), british isles (hanko 1983; coppins 2002; chambers et al. 2009), the czech republic (hanko 1983), estonia (randlane et al. 2008), fennoscandia (hanko 1983; santesson et al. 2004), italy (nimis and martellos 2008), slovenia (suppan et al. 2000). the species has also been recorded from macaronesia, north and south america and asia (dibben 1980; chambers et al. 2009; pišút 2009). in poland this species is rather rare and it is here confirmed only from north-eastern part of poland, the carpathians and one old locality in the sudetes (see hanko 1983) (fig. 1). specimens examined. poland. bg 30 – równina augustowska plain, puszcza augustowska forest, w of sucha rzeczka village, forest section no. 199-200, on alnus glutinosa, 11 sept. 1986, leg. s. cieśliński (ktc). bg 92 – wysoczyzna białostocka plateau, puszcza knyszyńska forest, budzisk nature reserve, on fraxinus excelsior, 5 aug. 1994, leg. s. cieśliński (ktc). cg 02 – puszcza knyszyńska forest, forest section no. 182, on carpinus betulus, 9 sept. 1987, leg. i. & k. toborowicz (ktc). cg 03 – puszcza knyszyńska forest, stare biele nature reserve, on tilia cordata, 4 aug. 1994, leg. s. cieśliński (ktc). cg 55 – równina bielska plain, puszcza białowieska forest, białowieża national park, forest section no. 256, on tilia cordata, 18 oct. 1989 and 5 june 1990, leg. s. cieśliński (ktc); ibidem, on tilia cordata, 10 may 1987 236 m. oset and m. kukwa and 29 oct. 2003, leg. k. czyżewska (lod–l 7504, 11836) and on carpinus betulus, 29 oct. 2003, leg. k. czyżewska (lod–l 11830); between forest sections nos 287d and 317b, on tilia cordata, 1982, leg. s. cieśliński & z. tobolewski (ktc); forest section no. 287, on tilia cordata, 1982, leg. s. cieśliński & z. tobolewski (kram-l 31318); forest section no. 341c, on tilia cordata, 1984, leg. s. cieśliński & z. tobolewski (ktc); forest section no. 400, on tilia cordata, 27 march 2001, leg. m. kukwa 210 (ugda); forest section no. 317d, on tilia cordata, 1982, leg. s. cieśliński & z. tobolewski (ktc). cg 56 – równina bielska plain, puszcza białowieska forest, białowieża national park, forest section no. 246, on tilia cordata, 1985, leg. s. cieśliński, z. tobolewski (ktc); cg 64 – puszcza białowieska forest, forest section no. 342b, on tilia cordata, 1979, leg. s. cieśliński & z. tobolewski (ktc); vicinity of topiło village, by perebel stream, forest section no. 599, on carpinus betulus, 12 may 2006, leg. m. kukwa 5098 (ugda); forest section no. 244b, on tilia cordata, 1984, leg. s. cieśliński & z. tobolewski (ktc); forest section no. 604b, on carpinus betulus, 1982, leg. s. cieśliński & z. tobolewski (ktc). ge 50 – high tatra mts, dolina roztoki valley, by roztoka stream, on alnus sp., july1964, leg. j. bystrek (lbl). ge 51 – rów podtatrzański depression, łysa polana settlement, on alnus sp., 16 may 1965, leg. j. bystrek (lbl, two specimens). poorly localized specimen. białowieża national park, s.coll. (lbl). additional specimens examined. austria. radstädter tauren, salzburg, an abies in einem feuchten bergwald zwischen oberund untertauren, ± 1240 m, oct.1963, leg. th. schauer, poelt & steiner, lich. alp. 222, sub p. multipuncta (kram-l 23765, lbl). conclusions the study has shown, that the secondary lichen metabolites are invaluable taxo-• nomic characters, that helps to evaluate the occurrence and distribution of at least some lichens. pertusaria multipuncta• and p. ophthalmiza are morphologically very similar and visually almost indistinguishable, but they differ in secondary chemistry. the analyses of lichen substances proved that p. multipuncta was mostly misidentified, and at present it is known from only one locality in gorce mts; most of remaining records of this species belong to p. ophthalmiza. fig. 1. distribution of pertusaria multipuncta and p. ophthalmiza in poland in the atpol grid square system; empty circle – locality of p. multipuncta, black circles – localities of p. ophthalmiza based on revised specimens; square – historical record of p. ophthalmiza from hanko (1983). the lichen genus pertusaria 237 p. ophthalmiza• appears to occur in well preserved natural ecosystems in north eastern poland and tatra mts. surprisingly, it was not found in bieszczady mts, the mountain range with suitable habitats for this species. acknowledgements. we would like to thank curators of all herbaria for the loan of material. we are indebted to an anonymous reviewer for suggestions on the manuscript. this work was supported by synthesys grants no. gb-taf-1013. references archer a.w. 1997. the lichen genus pertusaria in australia. biblioth. lichenol. 69: 1–249. chambers s. p., gilbert o. l., james p. w., aptroot a., purvis o. w. 2009. pertusaria dc. 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(por. hanko 1983; fałtynowicz 2003; kossowska 2008). na podstawie budowy morfologicznej i składu wtórnych metabolitów porostowych analizie poddano dwa gatunki z tego rodzaju, p. multipuncta (turner) nyl. i p. ophthalmiza (nyl.) nyl., które charakteryzują się podobną morfologią plech, ale różnią się chemizmem. p. multipuncta, podobnie jak p. ophthalmiza, cechuje się najczęściej cienką, gładką, niekiedy spękaną, biało-szarą plechą. owocniki, zwykle pojedyncze, są zagłębione w brodawkach plechy przypominających soralia, a tarczki są płaskie lub lekko wypukłe i przyprószone ziarenkowatymi sorediami. worki są jednozarodnikowe. gatunki te różnią się chemicznie. p. multipuncta reaguje na czerwono z pd i zawiera kwas fysodalowy (często także protocetrariowy), natomiast p. ophthalmiza nie reaguje z pd i zawiera kwasy tłuszczowe (hanko 1983; chambers et al. 2009). dotychczas z terenu polski częściej notowana była p. multipuncta (fałtynowicz 2003), p. ophthalmiza zaś była podana tylko raz przez hanko (1983). gatunek ten jednak nie został ujęty na liście porostów polski, a pierwsze dane o jego występowaniu na terenie kraju w polskiej literaturze lichenologicznej pojawiły się w pracy kukwy et al. (2008). przeprowadzone badania wykazały, że większość notowań p. multipuncta z polski należy do p. ophthalmiza lub do innych gatunków porostów. p. multipuncta została potwierdzona na terenie naszego kraju tylko z jednego stanowiska w gorcach. rozmieszczenie obu gatunków w polsce przedstawia rycina 1. 2014-01-01t11:51:36+0100 polish botanical society entoloma albotomentosum (agaricales, basidiomycota), a species new to poland marek halama museum of natural history, wrocław university sienkiewicza 21, pl-50-335 wrocław, marhalam@biol.uni.wroc.pl halama m.: entoloma albotomentosum (agaricales, basidiomycota), a species new to poland. acta mycol. 46 (2): 129–136, 2011. the first record of entoloma albotomentosum noordel. & hauskn., a member of the subgenus claudopus and section claudopus, is reported from poland. a full description and illustration of the species based on polish specimens are given and its taxonomy, ecology, and general distribution are also provided. key words: entolomataceae, entoloma sect. claudopus, pleurotoid habit, distribution, oleśnica plain, polish mycobiota introduction the genus entoloma (fr.) p. kumm. – the second largest genus of agaricales (after cortinarius) is monophyletic (co-david et al. 2009) and represented by at least 1000 species on a worldwide scale (kirk et al. 2008). although species of the section clau­ dopus noordel. of the genus entoloma are cosmopolitan, their basidiomata are often overlooked by researchers because of their small size or unusual habitats. fungi from this section are generally characterized by a pleurotoid or omphalinoid, small basidiocarps and typically excentric, lateral or lacking stipe (noordeloos 2004). they are saprotrophic, usually growing on dead organic matter such as rotten wood, bark, and various debris of grasses and sedges, and also on the ground, but some have been found to be parasites of other fungi (cantharellus cibarius, c. lutescens, coriolus ver­ sicolor, xanthochrous perennis) and mosses (sphagnum, aulacomnium) (noordeloos 1992, 2004). just because of the pleurotoid (or crepidotoid) growth form, a lot of these species are sometimes placed in separate genus caludopus gillet. (horak 1980; largent et al. 2011). however, most authors consider the genus caludopus artificial rather than natural, and place these fungi in subgenus claudopus and section acta mycologica vol. 46 (2): 129–136 2011 130 m. halama claudopus within the genus entoloma (noordeloos 1992, 2004), a concept favoured in this paper. out of 10 species belonging to the section claudopus in europe, so far no more than 5 species have been reported to occur in poland: e. byssisedum (pers.: fr.) donk, e. depluens (batsch: fr.) hesl., e. jahnii wölfel & winterhoff, e. ollare ludwig & röbig and e. parasiticum (quél.) kreisel (wojewoda 2003; nita, bujakiewicz 2006; mleczko, ociepa 2007; nita, bujakiewicz 2007; kujawa 2009; nita, stefaniak 2010; stefaniak 2010). during a period of damp summer weather in august 2010, unusual pleurotoid basidiocarps were found growing on dead, fallen reedgrass leaves in the area of post-agricultural land at domaszowice, oleśnica plain (sw poland; atpol: ec–55; fig. 1). laboratory studies showed that they represent a very interesting member of the claudopus section, entoloma albotomentosum noordel. & hauskn. according to the author’s knowledge, this species has not been reported from poland to date and the present collection is the first record in the country. e. albotomentosum is a rare species in many countries of europe and very variable. for that reason a detailed description and illustrations are presented to facilitate identification of this fungus from further localities. material and methods the study is based on material gathered by the author in 2010 during the mycological research in the south-western poland. the material was gathered within one locality. the samples were documented with colour photographs (partly shown on fig. 2). the description of macroscopic features is based on fresh material, comprising 2 collections, 10 basidiomata in all stages of development. the microcharacters fig. 1. the locality of entoloma albotomentosum in poland (based on a 100 km atpol grid) (a), and in the opole province (b): 1 – border of the country, 2 – administrative boundaries of the opole province, 3 – locality of the species. fig. 2. basidiocarps of entoloma albotomentosum recorded at domaszowice (coll. 14.08.2010; photo by m. halama). fig. 3. basidiospores (a), basidia (b), and hyphae of pileipellis (c) of entoloma albotomentosum recorded at domaszowice (coll. 14.08.2010). entoloma albotomentosum, new to poland 131 of 2 recorded basidiomata (from 2 collections) were observed and measured under a light microscope at 1500× (basidiospores) and 800× (other features). for microscopic observations, dried pieces of basidiomata were placed in 5% nh4oh for about 1 minute, then transferred to tap water until they become pliable. free-hand sections of the rehydrated pieces of basidiomata were examined in 5%nh4oh, congo red and in phloxine (in 1%nh4oh). amyloidy was tested with melzer’s reagent. morphological measurements were made and are presented according to the method used by breitenbach & kränzlin (1991). the 95% population limits for the mean were calculated and the lower and upper limits are presented. the minimum and maximum dimensions are given in parentheses additionally. the ratio of basidiospore length to its width (q) was calculated. the length of basidia was measured excluding sterigmata. all statistical analyses were performed using statistica software (version 8, statsoft). terminology of morphological and anatomical features has been adopted mainly from vellinga (1988) and noordeloos (1992). reported size of basidiospores, basidia and pileipellis hyphae were based on 101, 31 and 31 measurements, respectively. microphotographs were taken using nikon ds.-fi1 digital camera. the voucher specimens of e. albotomentosum have been deposited in wrsl (museum of natural history, wrocław university, wrocław, poland). results and discussion entoloma albotomentosum noordel. & hauskn. z. mykol. 55(1): 32–33. 1989 – entolomataceae, agaricales, agaricomycetidae, agaricomycetes, basidiomycota, dikarya, fungi (hibbett et al. 2007). icones: noordeloos, hausknecht (1989: 32-33); noordeloos (1992: tab. 72c; 2004: 13571358, fot. 324). basidiomata most often eccentric stipitate (fig. 2). pileus 1.5-5.4 mm broad, 0.51.5 mm deep, irregularly circular, initially convex to applanate with usually decurved margin, in old stage often somewhat applanate-depressed, and with undulating, ± grooved marginal zone, not clearly hygrophanous, when expanding usually translucently striate, white, turning pinkish because of the colour of the lamellae, silvery white fibrillose when young, then, especially at centre with bundles of erect, agglutinated white hairs. lamellae, l = 5-14, very distant, adnate to subdecurrent or decurrent, often thickened, vein-like, forked and sometimes anastomosing, white then pink with concolorous, entire edge. stipe 0.6-2 × 0.2-0.4 mm, weakly to distinctly eccentric, often curved, white, covered with fine hairs, then glabrous, except for the base. context very thin and delicate. smell imperceptible. basidiospores (7.23) 8.869.05 (12.14) × (5.89) 6.95-7.09 (8.87) μm, q = (0.97) 1.27-1.29 (1.64), 4-6-angled in side view, weakly pigmented, pale yellow (in 5%nh4oh), with a great guttule, inamyloid. basidia (25.14) 28.86-30.55 (36.39) × (9.85) 11.75-12.64 (14.50) μm, narrowly clavate, hyaline or containing small granules while immature, mostly with 4 sterigmata, without basal clamp. lamella edge fertile. pileipellis: a cutis with transitions to a trichoderm, made up of cylindrical to inflated, (3.41) 7.32-8.13 (12.01) μm wide hyphae; pigment absent. clamps absent (fig. 3). 132 m. halama material examined: 1. poland, domaszowice (oleśnica plain), idle rural land with scattered betula and calamagrostis, rubus and solidago in the field layer: on damp rotten leaf-sheaths and debris of herbaceous plants (calamagrostis sp.), 14.08.2010, leg. m. halama, wrsl; 2. poland, domaszowice (oleśnica plain), idle rural land with scattered betula and calamagrostis, rubus and solidago in the field layer: on damp rotten debris of herbaceous plants (calamagrostis sp. ?), 17.08.2010, leg. m. halama, wrsl. entoloma albotomentosum was described from austria (irnfritz: teichholz) on rotten debris of grasses (calamagrostis epigejos) and sedges (carex) (noordeloos, hausknecht 1989). since then it was rarely found in several other european countries, viz. the netherlands, liechtenstein, germany, denmark, england, scotland and norway (krieglsteiner 2003; legon et al. 2005; arnolds, veerkamp 2008; noordeloos 2008). this species is distinctive among the european members of the section claudopus mainly with its unique ecological requirements (occurrence on rotten leaf-sheaths and debris of grasses and sedges) (noordeloos 2004), therefore some accessible data relating to its incidence on wood substrata seem to be doubtful (musumeci 2005; dechaume 2010). this fungus is regarded to be confined to damp woodlands or marshy places at edges of ponds (noordeloos 1992; krieglsteiner 2003; legon et al. 2005). the recorded locality of e. albotomentosum in oleśnica plain offered some different moisture conditions, as the explored habitat was rather dry and never flooded. however, the collection from this site was made after prolonged rain, in places shaded by birches, on very damp debris of calamagrostis. the habit of the basidiocarps of e. albotomentosum oscillates between pleurotoid with reduced, lateral or lacking stipe, to omphalinoid with well-developed, sometimes central stipe (noordeloos 1992, 2004). the polish specimens of e. albo­ tomentosum agree well macroscopically with the descriptions of noordeloos & hausknecht (1989) and noordeloos (1992, 2004, 2008). microscopically, this collection shows some little differences in the spore size from the measurements presented by earlier authors (tab. 1), but these discrepancies are probably attributable to the different sampling sizes. e. albotomentosum is similar in outward appearance to the species more widespread in europe – entoloma jahnii wölfel & winterh., but is well separated morphologically as well as ecologically. it is distinguished primarily by the lack of clamps in all parts of basidiocarps and the lack of capitate pileoand caulocystidia, and also smaller basidiospores. moreover, the habitats of the two species are different, because e. jahnii is found in hidden places, such as the inner side of rotten bark and the underside of rotten wood of deciduous tree stumps and logs (wölfel, winterhoff 1993; noordeloos 2004; nita, stefaniak 2010). some superficial similarity has also entoloma alliodorum esteve-rav., e. horak & a. ortega with its white colour, but it differs generally by the more omphalioid habit (more distinct stipe), crowded lamellae, somewhat smaller, heterodiametrical basidiospores, strong smell of garlic and growing on mosses and rotten debris (esteve-raventós, ortega 2003; noordeloos 2004). there are also others non-european species, morphologically resembling e. albotomentosum in their pleurotoid, white basidiocarps, similarly shaped basidiospores, and lacking clamp connections. entoloma pandanicola (e. horak) noordel. & co-david (from papua new guinea) can be distinguished primarily by habitat – rotting pandanus leaves, farinaceous smell, adnexed to free lamellae and shorter (7-8 μm) basidiospores (horak 1980; co-david et al. 2009), while entoloma albotomentosum, new to poland 133 claudopus rupestris largent & abell-davis (from australia) can be differentiated by its smaller, glistening-sticky, opaque, not translucently striate pileus (1-4 mm broad), adnexed – more ventricose lamellae, shorter (6.5-9.2 μm) basidiospores, presence of pileocystidia, and habitat it grows on undersurface of small granite rocks (growing on thin to nearly non-existent soil layer with minute mosses) (largent et al. 2011). e. albotomentosum reminds also small crepidotus or even clitopilus species. some representatives of the mentioned genera are also found on debris of herbaceous plants (including the rotten leaf blades of grasses), however, e. albotomentosum can be easily distinguished from these crepidotoid or pleurotoid fungi by their basidiospores, which are globose, ovoid, ellipsoid, cylindrical to fusiform (smooth to rugose or verrucose or finely spiny) in crepidotus, and ellipsoid or ± amygdaloid and angular in polar view in clitopilus. table 1 comparison of the spore characteristics of e. albotomentosum according to different authors authors spore length (μm) spore width (μm) spore length/width ratio noordeloos & hausknecht (1989), noordeloos (1992, 2008) 9.0-12.5 6.5-8.0 1.1-1.5 present studies 7.23-12.14 95% population limits for the mean (8.86-9.05) 5.89-8.87 95% population limits for the mean (6.95-7.09) 0.97-1.64 95% population limits for the mean (1.27-1.29) recognized as a rare species, e. albotomentosum is red-listed in austria (krisaigreilhuber 1999), denmark (national environmental research institute 2011), and in the netherlands (arnolds, veerkamp 2008). conclusion the infrequency of the species may be due to its small, inconspicuous basidiocarps, which are often hidden under mats or tufts of grasses and sedges, and therefore they are very difficult to find. that is also a possible reason why e. albotomentosum has only been recorded recently in poland. this species is not well enough studied to make a statement about its rarity in poland and further field studies are needed to determine a real condition and distribution of the species here. acknowledgements. i am indebted to the anonymous reviewer for valuable comments on the manuscript. i would like to thank thomas læssøe for his kind help with completing information on european distribution of e. albotomentosum. i also thank katarzyna dunaj for her assistance in collecting material. 134 m. halama references arnolds e., veerkamp m. 2008. 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(in:) z. mirek (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków, 812 pp. wölfel g., winterhoff w. 1993. entoloma jahnii, ein neuer holzbewohner. österreichische zeitschrift für pilzkunde 2: 11–14. entoloma albotomentosum (agaricales, basidiomycota), gatunek nowy dla polski streszczenie w polsce stwierdzono dotychczas występowanie 5 przedstawicieli sekcji claudopus, rodzaju entoloma (dzwonkówka), tj.: e. byssisedum, e. depluens, e. jahnii, e. ollare i e. parasiticum. grzyby te charakteryzują się z reguły drobnymi owocnikami typu boczniakowatego (pleurotoidalnego) lub pępówkowatego (omfaloidalnego). są wśród nich gatunki saprotroficzne, rozwijające się na różnych, butwiejących szczątkach roślin wyższych lub pojawiające się bezpośrednio na ziemi. należą tu również gatunki patogeniczne, rozwijające się na owocnikach innych grzybów lub związane z obecnością mszaków. w trakcie obserwacji mikologicznych, przeprowadzonych w sierpniu 2010 roku w miejscowości domaszowice (równina oleśnicka), znaleziono przypadkowo na terenie nieużytku porolnego kilka bardzo drobnych, białawo zabarwionych, pleurotoidalnych owocników, zasiedlających martwe szczątki traw (głównie butwiejące liście trzcinnika calamagrostis sp.). badania laboratoryjne wykazały, iż reprezentują one nowy dla mikobioty polski gatunek dzwonkówki – entoloma albotomentosum noordel. & hauskn. (proponowana polska nazwa: dzwonkówka ciżmówkowata). e. albotomentosum została opisana w 1989 roku z terenu austrii. w najnowszych opracowaniach takson ten ujęty jest w podrodzaju i sekcji claudopus. gatunek ten cechuje się wytwarzaniem drobnych, niepozornych, biało zabarwionych i dość zmiennych w budowie makroskopowej owocników, które mogą mieć formę boczniakowatą (z trzonkiem wykształconym bocznie, zredukowanym bądź niewykształconym) lub pępówkowatą (z trzonkiem wyraźnie wykształconym i niekiedy centralnie osadzonym). w młodym stadium owocniki te zwykle przypominają do złudzenia niewielkich przedstawicieli rodzaju ciżmówka (crepidotus). według danych literaturowych e. albotomentosum pojawia się przede wszystkim w gęstych zaroślach traw i turzyc oraz na brzegach stawów, gdzie tworzy owocniki na butwiejących szczątkach traw i turzyc. rozmieszczenie tego gatunku jest słabo poznane. do chwili obecnej stwierdzono jego występowanie jedynie w europie, gdzie (oprócz austrii) znany jest on z rozproszonych stanowisk, zlokalizowanych w holandii, księstwie lichtenstein, niemczech, danii, anglii, szkocji i norwegii. specyficzne preferencje siedliskowe oraz niewielkie rozmiary owocników e. albotomento­ sum są prawdopodobnym powodem rzadkości notowań tego gatunku w terenie. 136 m. halama dokładniejsze poszukiwania dają szansę na znalezienie dalszych stanowisk tego grzyba, m. in. w polsce. w prezentowanej pracy przedstawiono szczegółowy opis i ilustracje cech zebranych owocników e. albotomentosum. przedstawiono również informacje na temat taksonomii, ekologii i rozmieszczenia tego gatunku. 2014-01-01t23:44:17+0100 polish botanical society a new record of the rare earthstar geastrum berkeleyi from the świętokrzyskie mts justyna jaworska department of botany, institute of biology, jan kochanowski university świętokrzyska 15, pl-25-406 kielce, j.jaworska.kielce@gmail.com jaworska j.: a new record of the rare earthstar geastrum berkeleyi from the świętokrzyskie mts. acta mycol. 46 (1): 75–81, 2011. the article presents the third record of geastrum berkeleyi massee in poland. for a long time the only known polish locality of the species was in kobylnica near poznań, where g. berkeleyi was recorded in 1934. the fungus was found for the second time as recently as 2007 in rąbiń (kościan county). in 2009 this earthstar species was recorded in milechowy reserve near chęciny, in the western part of the świętokrzyskie mts (square atpol ee 72). in the last mentioned locality, fruit-bodies of this rare fungus were recorded in a thermophilous forest habitats and also in a xerothermic anthropogenic shrublands, on a sandy calcareous soil. a comparison of diagnostic features of geastrum berkeleyi var. berkeleyi and g. berkeleyi var. continentale is also given. key words: distribution, macrofungi, milechowy reserve, poland introduction the genus geastrum is represented in poland by 17 species (wojewoda 2003). considering a characteristic morphological structure of fruitbodies they are relatively easy to recognize. differences between particular species concern above all the sizes of fruitbodies, the shape of an exoperidium, the means in which an endoperidial body is attached to the exoperidium, the structure of a peristome, the habitat requirements and some microscopic features, especially the structure and the size of basidiospores. all species of the genus geastrum are either rare or very rare and only a few, such as geastrum quadrifidum dc.: pers., g. rufescens pers., g. fimbriatum fr. and g. minimum schwein. can be considered as relatively frequently occurring. since 2004 all species of this genus are strictly protected in poland (rozporządzenie 2004). acta mycologica vol. 46 (1): 75–81 2011 a new record 77 results species description geastrum berkeleyi massee annals of botany 4: 79.1889 (as geaster) (syn.: geastrum pseudostriatum hollós (1901), geastrum hollosii v.j. staněk (1958) systematic arrangement: geastraceae, lycoperdales, gastromycetes (jülich 1984); lycoperdales, basidiomycetes (rudnicka-jezierska 1991); lycoperdales, basidiomycetes, basidiomycota (hawksworth et al. 1995); lycoperdales, hymenomycetidae, hymenomycetes, basidiomycota (sunhede 1997); phallales, agaricomycetidae, basidiomycetes, basidiomycota (kirk et al. 2001); geastrales, phallomycetidae, agaricomycetes, basidiomycota, fungi (kirk et al. 2008). fruit-bodies typical for genus geastrum. immature fruit-bodies are closed and subglobose, 1-3.5 (4) cm in diameter. exoperidium consists of three layers, and in the unexpanded fruit-bodies closely invests endoperidium. the exterior surface of young basidioma covered with plant debris and soil particles held by the adnate, light ochre-brown and sometimes rose tinged mycelial layer. as the fruit-bodies ripens the mycelial coating tends to flake away and the smooth, not shiny fibrillar layer is uncovered. exoperidium splits open from the tip to about the middle in a stellate fashion divided into 4–10 triangular flaps (6-7 flaps in the specimens collected in milechowy reserve). due to swelling of pseudoparenchymal cells of exoperidium, the triangular flaps curve outwards and make contact with the soil, lifting the inner endoperidial layer of fruit-bodies into the air (fig. 2). in such cases the flaps diameter reaches 4–10 cm. the inner, fleshy layer of exoperidium is breakable and redish-brown or ochre coloured. endoperidium globose or slightly flattened, usually has diameter from 1 to 3 cm and it is seated upon a short but broad pedicel, which can be usually clearly seen only in dried specimens. indistinct apophysis is present, but also is frequently clearly seen in dry specimens only. endoperidium is ochre-brown, dark brown, grey-brown coloured, and in older fruit-bodies is grey in colour. its surface is coarsely verrucose, except for a smooth, circular area surrounding peristome. peristome conical, distinctly plicate with about 15-18 folds, grey-brown to brown. its opening is crest-like frayed or bluntly ended. gleba dark brown when mature with chocolate tinge at times. capillitial hyphae yellow-brown, unbranched, up to 13 μm thick. basidiospores globose, distinctly verrucose, olivaceous-brown (in h20, koh) (fig. 3), 4.75-6.10 μm in diameter. spores measurements for this species given by different authors are: 4.5-6.75 μm (pilát 1958), 4.5-6.7 μm (jülich 1984), 4.5-6 (-7.5) μm (rudnickajezierska 1991), 5.5-7 μm (hansen, knudsen 1997). the sizes of investigated spores from the fruit-bodies found in milechowy reserve are close to those given by the above authors. material examined. świętokrzyskie mts milechowy reserve near chęciny, ca 230 m a.s.l.: in young planted pine forest, on sandy, calcareous soil, 07.09.2009. the collected specimens have been deposited in the fungarium of the mathematics and nature faculty, jan kochanowski university, kielce (ktc 4305). 78 j. jaworska taxonomical remarks. geastrum berkeleyi is well characterized by arched, nonhygroscopic exoperidial flaps, clearly roughened – coarsely verrucose endoperidium, and distinctly plicate, conical peristome. macromorphological features of the examined specimens are in good agreement with the previous description of g. berkeleyi (e.g., rudnicka-jezierska 1991). also micromorphological characteristics correspond to those given in mycological literature, although it is worth emphasizing here that various authors have reported different measurements of spores, i.e.: 4.5-6.75 μm (pilát 1958), 4.5-6.7 μm (jülich 1984), 4.5-6 (-7.5) μm (rudnicka-jezierska 1991), 5.5-7 μm (sunhede 1997). staněk (1958) distinguished two varieties of geastrum berkeleyi, typical – var. berkeleyi massee and var. continentale v.j. staněk. both varieties differ from each other in the following features: apophysis appearance, nature of ornamentation of basidiospores, and width of capillitial hyphae (tab. 1). fruit-bodies collected in the milechowy reserve correspond to the descripition of g. berkeleyi var. continentale in this respect. geastrum berkeleyi shows particular similarities to a few other species belonging to this genus. g. campestre morgan. (syn. g. pedicellatum (batsch) dörfelt & müll.-uri), g. kotlabae v.j. staněk and g. pectinatum pers.: pers. are the other taxa having a plicate, conical peristomes and except for the third mentioned – mealy covered or rough surface of endoperidia. however, g. campestre is marked by its hygroscopic rays, more persistent pseudoparenchymatous layer of the exoperidium and smaller fruit-bodies (usually 6-20 mm in diameter) as well. g. kotlabae is also distinguished from g. berkeleyi by its strongly hygroscopic rays and more persistent pseudoparenchymatous layer of the exoperidium as well as additionally by the sessily seated endoperidial bodies. the surface of exoperidial bodies of g. kotlabae is also somewhat different. it is farinose in young fruit-bodies, and usually glabrous in older basidiomata. the features differentiating g. pectinatum from g. berkeleyi can mainly be seen in the morphology of pedicel and apophysis. g. pectinatum is characterized by having a long pedicel, and usually radially wrinkled – sulcate or striate apophysis, while the pedicel of g. berkeleyi is low and broad, and the apophysis is smooth. also, g. pectinatum differs from g. berkeleyi in smooth surface of endoperidium. table 1 comparison of diagnostic features of geastrum berkeleyi var. berkeleyi and g. berkeleyi var. continentale feature geastrum berkeleyi var. berkeleyi geastrum berkeleyi var. continentale apophysis weakly developed (poorly visible) well developed width of capillitial hyphae up to 13 μm does not exceed 10.5 μm spores spherical, up to 6 μm in diameter spherical, 4.5-6.2 μm ornamentation of spores up to 15 verrucae along the circumference 12 – 16 verrucae along the circumference a new record 79 general distribution and ecology geastrum berkeleyi is a rare species. it is known from sparse locations in asia and europe. in asia it has been reported from japan and china (kasuya et al. 2009). in europe g. berkeleyi occurs in a dozen or so countries, including austria, the czech republic, denmark, estonia, france, germany, great britain, hungary, the netherlands, poland, slovakia, spain, sweden and turkey (arnolds, veerkamp 2008; gbif 2009; dörfelt 1985; honorubia et al. 1980; jülich 1984; lilleleht 1998; lizoň 2001; sunhede 1989, 1997). in most european countries where the species was noted it appears on the “red lists” of threatened fungi: in austria is listed as endangered by extinction (“1” category), in the czech republic and denmark as critically endangered (“cr” and “e” category), in estonia as vulnerable (“v” category), in germany as a rarity (“r” ca tegory), in the netherlands as susceptible and rare (“ge(z)” category), in slovakia as vulnerable (“vu” category) and in great britain and sweden as endangered (“en” category) (evans 2006; wojewoda 2003). geastrum berkeleyi was considered to be “extinct and probably extinct” in the last edition of the “red list of macrofungi in poland” (wojewoda, ławrynowicz 1992, 2006). certainly, the species is not extinct in poland. currently, it occurs at least in two localities in the area and therefore the status of the taxon should be changed to endangered. the detailed ecology of g. berkeleyi in northern europe was addressed by sunhede (1989). other valuable data on the ecology of the earthstar in europe were also provided by staněk (1958), dörfelt (1985) and kreisel (1987). according to these authors, g. berkeleyi occurs mostly on sun-warm sites, in coniferous and deciduous forests, groves and tree plantations (with picea, pinus, acer, carpinus, corylus, crataegus, fagus, fraxinus, juniperus, prunus, quercus and tilia), on well drained, base-rich ground, especially on calcareous soils. it is also known from pine and xerothermic shrubs, syringa-shrubberies and open sites. most finds of fresh fruit-bodies of the species have been made in september and october, but records from the end of august and beginning of december are also known. basidocarps of g. berkeleyi can grow scattered or in clusters, sometimes in bows or fairy rings (sunhede 1989). until now g. berkeleyi was reported from two various habitats in poland, i.e.: from a margin of spruce grove (teodorowicz 1939) and a riparian ash forest with undercrop of crataegus and prunus (kujawa, gierczyk 2010). the new, presented here locality of g. berkeleyi recorded in milechowy reserve was observed on sandy, calcareous soil in thermophilous forest habitat. the floristic composition of the plant habitat, in which several fruit-bodies of the species were found is as follow: tree layer with 70% density, shrub layer with 15% density, herb layer with 40% cover, moss layer with 30% cover. the tree layer consists only of pinus sylvestris (4.4). in the shrub layer occur: lonicera xylosteum (1.1), quercus petraea (1.2), juniperus communis (1.1), carpinus betulus (+). in the herb layer mainly occur: festuca rubra (3.1), fragaria vesca (1.1), hieracium pilosella (+.2), thymus serpyllum (1.2), chimaphila umbellata (1.1), melam pyrum pratense (2.2), vaccinium myrtillus (+.2), galium mollugo (+.2), veronica officinalis (1.1). the moss layer is represented mostly by entodon schreberi (3.1) and hylocomium splendens (3.1). 80 j. jaworska acknowledgements. i’m very grateful to prof. janusz łuszczyński and prof. andrzej massalski for helpful discussion and comments on the manuscript. references arnolds e., veerkamp m. 2008. basisrapport rode lijst paddenstoelen. nederlandse mycologische vereniging, utrecht, 295 pp. braun-blanquet j. 1964. pflanzensoziologie. wien-new york. dörfelt h. 1985. die erdsterne: geastraceae und asteraceae. die neue brehm-bücherei, a. ziemsen verlag, wittenberg, 108 pp. evans s. 2006. the red data list of threatened british fungi: a preliminary assessment (version 1.0). british mycological society: published on the web site: http://www. fieldmycology.net/download/ [accessed: 11-2010]. gbif 2009. biodiversity occurrence data provided by: steiermärkisches landesmuseum joanneum herbarium gjo, university of vienna, institute for botany herbarium wu (accessed through gbif data portal, data.gbif.org, 2009-11-02). hawksworth d.l., kirk p.m., sutton b.c., pegler d. 1995. ainsworth & bisby’s dictionary of the fungi. 8th ed. intern. mycol. inst., university press, cambridge, 616 pp. honrubia garcía m., calonge f.d.d., demoulin v., moreno g., llimona x. 1980. aportación al conocimiento de los hongos del se. de españa vi: esclerodermatales lycoperdales, nidulariales, phallales, hymenogasterales, podoxales (gasteromycetes, basidiomycetes). anales de la universidad de murcia: ciencias 38 (1/4): 101–132. index fungorum. 2009. published on the web site: http://www. indexfungorum.org/names/names.asp [accessed: 11-2009]. jülich w. 1984. die nichtblätterpilze, gallertpilze und bauchpilze. aphyllophorales, heterobasidiomycetes, gastromycetes. 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(in:) k. zarzycki, w. wojewoda, z. heinrich (eds). lista roślin zagrożonych w polsce (wyd. 2). list of threatened plants in poland (2nd ed.). w. szafer institute of botany, polish academy of sciences, kraków: 27–56. wojewoda w., ławrynowicz m. 2006. red list of the macrofungi in poland. czerwona lista grzybów wiel-wielkoowocnikowych w polsce. (in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. w. szafer institute of botany, polish academy of sciences, kraków: 53–70. nowe stanowisko rzadkiego gwiazdosza geastrum berkeleyi z gór świętokrzyskich streszczenie w polsce dotychczas znane były tylko 2 stanowiska geastrum berkeleyi. po raz pierwszy znaleziono go w roku 1934 w kobylnicy koło poznania (teodorowicz 1939). jego drugie stanowisko stwierdzono w 2007 r. w rąbinie w województwie wielkopolskim (kujawa 2009; kujawa, gierczyk 2010). w 2009 r. znaleziony został w rezerwacie milechowy koło chęcin, w zachodniej części gór świętokrzyskich. jest to trzecie jego znane stanowisko w polsce. ten rzadki gatunek występuje w ciepłych lasach, często pochodzenia antropogenicznego oraz w kserotermicznych zaroślach i na piaszczystych glebach wapiennych. gatunek znany w europie i azji. w europie odnotowano jego występowanie w austrii, republice czeskiej, danii, estonii, francji, niemczech, wielkiej brytanii, holandii, słowacji, hiszpanii, szwecji, turcji oraz na węgrzech (arnolds, veerkamp 2008; gbif 2009; dörfelt 1985; honorubia et al. 1980; jülich 1984; lilleleht 1998; lizoň 2001; sunhede 1989, 1997). w większości krajów należy do grzybów ujętych na czerwonych listach gatunków zagrożonych. 2014-01-01t11:51:58+0100 polish botanical society lichens of the holy hill orthodox sanctuary in grabarka (ne poland) anna matwiejuk department of botany, institute of biology, university of bialystok świerkowa 20b, pl-15-950 białystok, matwiej@uwb.edu.pl matwiejuk a.: lichens of the holy hill orthodox sanctuary in grabarka (ne poland). acta mycol. 43 (1): 105–111, 2008. the holy hill grabarka is one of the most important orthodox sanctuaries in poland. the sanctuary is situated in podlasie region between bug and narew rivers. it grew in the shade of well developing (in the first centuries of the second millennium) towns mielnik and drohiczyn. the most striking thing is that the church is surrounded by a forest of thousands of crosses brought by pilgrims. the study present 64 species of epiphytic, epixylic, epilythic and epigeic lichens. among 64 lichen species 11 are considered to be threatened in poland. key words: lichenized fungi, biota, holy hill sanctuary introduction the holy hill grabarka is the most significant orthodox sanctuary in poland. the sacred place is situated 12 km away from siemiatycze, 0,5 km from the grabarka village in podlaskie province, siemiatyckie district, commune nurzec-stacja. the holy hill is found on wysoczyzna drohicka high plain, in the south part of nizina północnopodlaska lowland (kondracki 1994). in the close neighborhood there is the river moszczona, which falls into the river bug, 7 km further to the south. the surrounding woods had been formerly included in the mielnicka forest, which is known as the lasy nurzeckie forests. study area the holy hill grabarka acquired its fame for a miracle which occurred in the beginning of 1710 in the time of cholera epidemic, which decimated the population of podlasie. as legend says, at that time, an old man from siemiatycze had a revelation according to which it was possible to be protected from the disease and be saved only on the grabarka hill. after the faithful from siemiatycze had arrived and placed there a cross, a miracle occurred. the sick started to recover and nobody died afterwards. when the news spread, people from that region arrived at the holy hill acta mycologica vol. 43 (1): 105–111 2008 106 a. matwiejuk in hundreds. in 1710, in summer, about 10 thousand people gathered in this place. in the same year, a wooden temple was erected there as gratitude of those who were saved. the holy grabarka hill is also known as the hill of crossesthe name comes from sacrificial crosses with epitaphs written in cyrillic alphabet, which are placed around the transfiguration orthodox church. the crosses which surround the small church have become the symbol of the holy hill. it is a real forest of crosses with narrow paths to walk along (radziukiewicz 2001). at present, approximately 10 thousand crosses tower on the hill – wooden, metal, stone, small and big. the crosses have been traditionally brought by the faithful for hundreds of years. the pilgrims place them next to the sanctuary throughout the year, however the most crosses occur the day before the transfiguration holiday (lechowski 2004). behind the amazing forest of crosses, a cemetery and buildings of the st. martha and st. mary are found. the nunnery was established in 1947 and the holy virgin patroness and succor orthodox church was erected in 1957. construction of stone wall to enclose the holy hill was begun in 1999 as a result of benediction of his eminence metropolitan sawa (radziukiewicz 2000). the objective of the present paper has been the floristic and ecological analysis of lichen biota of the holy grabarka hill and its surrounding for the purpose of presenting its variety considering the habitat conditioning. material and methods the investigations in the area of the holy hill orthodox sanctuary in grabarka and its surroundings were carried out in the years 2006-2007. the alphabetical list of lichen species has been compiled (tab. 1). several lichen species from the grabarka surroundings (stand 1182 alnus glutinosa over flow, sandy slope, pinus sylvestris) have been reported by cieśliński (2003a). the species confirmed by our own studies on that stand have been marked (also cieśliński 2003a). the species which have not been identified in the course of our own studies have been marked cieśliński (2003 a). the species have been named according to santesson et. al. (2004), genera bryoria and usnea according to bystrek (1986, 1994) and genus melanelia according to blancoet al. (2004). the lichen material has been deposited at the herbarium of the institute of biology, university of białystok. results 64 species of lichens, out of 35 genera have been identified on the area of the holy grabarka hill and its neighborhood. the most abundant are genera cladonia – 8 species, lecanora – 7, caloplaca – 4. the lichens occur in all basic morphological forms. crustose thalus species remarkably dominate (45%). there are less macrolichens – foliose (29%) and fruticose (25%). of the 64 lichen species identified in the holy hill orthodox sanctuary in grabarka and its environs, 11 species have been put on the red list of extinct and vulnerable lichens of poland (cieśliński et al. 2003), including 3 species in the endangered category – en (bryoria crispa, flavoparmelia caperata, pleurosticta acetabulum), 6 species in the vulnerable category – vu lichens of the holy hill 107 (cetraria islandica, parmelina tiliacea, porpidia rugosa, tuckermanopsis chlorophylla, usnea filipendula, u. hirta), 2 species in the category of near threatened – nt (evernia prunastri, hypogymnia tubulosa), as well as 2 species on the red list of lichens vulnerable in north-eastern poland (cieśliński 2003 b), including 1 species in the category en (flavoparmelia caperata), 1 – lc (porpidia rugosa). lichens which are protected by law constitute 27% of the total lichen biota, including strict environmental protection species – 20%, and partial protection – 6%. zones of protection within a radius of 50 m from the rim of the species locality should be specified in regard to usnea hirta and usnea filipendula (dz. u. nr 168, poz. 1765). epiphytic lichens. the most abundant are epiphytic lichens (31 species), out of which 11 are obligatory. they occur on the bark of pinus sylvestris, betula pendula, quercus robur, acer platanoides, alnus glutinosa and juniperus communis. considering floristic differentiation and abundance, the richest is lichen biota pinus sylvestris (11 species), acer platanoides and betula pendula (10 of each). fruticose species, generally perishing in poland – usnea hirta, u. filipendula, pseudeuernia furfuracea as well as foliose hypogymnia physodes occur in considerable amount on the bark of pine – and birch – trees. the lower parts of the trunks are mainly colonized by the lichens of cladonia genus (c. chlorophaea, c. coniocraea, c. fimbriata). the bark of numerous trunks, including the crown, is profusely overgrown with lichens thalus. 11 species, exclusively flavoparmelia caperata (cieśliński 2003a), lecanora carpinea, l. pulicaris, pleurostica acetabulum (cieśliński 2003b) were identified on the bark of alders growing on the stream. species composition of the lichens of pine – trees growing in the nearest woods is remarkabely scantier. common species such as hypogymnia physodes, hypocenomyce scalaris, cladonia coniocraea significantly dominate, and imshaugia aleurites and lecanora conizaeoides are found relatively often. epixylic lichens. these lichens are included in the second, regarding the number, habitat group. they are represented by 24 species, out of which only 3 (lecanora varia, parmelina tiliacea, placynthiella icmalea) are exclusive epixylic lichens. they mainly grow on the wooden crosses which surround the church. quite frequently they form big groups composed of a dozen or so – several dozens specimens. mainly epiphytic species occur among epixylic species. the most abundant are foliose thalus platisimatia glauca, parmelia sulcata, hypogymnia physodes, parmelina tiliacea, melanelixia fuliginosa, imshaugia aleurites as well as fruticose thalus pseudevernia furfuracea, evernia prunastri, usnea hirta, u. filipendula, cladonia fimbriata, c. coniocraea. wood left to rot after the cut down trees form the second breeding ground overgrown by epixylic lichens, such as hypogymnia physodes or imshaugia aleurites, lepraria sp., parmelia sulcata, placynthiella icmalea. epilithic lichens. 24 species have been identified, out of which 19 exclusive. lichens of the holy grabarka hill are connected with natural breeding ground (erratic blocks, stones, tombstones, stone wall) – 15 species, out of which caloplaca holocarpa, candelariella aurella, lecanora albescens, l. dispersa, physcia caesia, protoparmeliopsis muralis, xanthoria parietina and x. polycarpa grow also on calcium breeding ground, on concrete structures mainly (tombstones). 9 species were identified exclusively on breeding ground of calcium substrate. in the grabaraka vicinity, cieśliński (2003a) identified porpidia rugosa (individual without apothecium) on 108 a. matwiejuk ta b l e 1 lichens recorded on the holy hill in grabarka species substratum notes acarospora fuscata (schrad.) th. fr. erratic blocks, stone wall also cieśliński (2003a) amandinea punctata (hoffm.) coppins & scheid. trunk of acer platanoides bryoria crispa (mot.) bystr. trunk of pinus sylvestris caloplaca citrina (hoffm.) th. fr. concrete tombstones caloplaca decipiens (arnold) blomb. & forssell concrete tombstones caloplaca holocarpa (hoffm. ex ach.) a. e. wade stones, stone wall, concrete tombstones caloplaca saxicola (hoffm.) nordin concrete tombstones candelariella aurella (hoffm.) zahlbr. concrete tombstones, stone wall candelariella vitellina (hoffm.) müll. arg. concrete tombstones candelariella xanthostigma (ach.) lettau trunk of acer platanoides cetraria aculeata (schreb.) fr. soil also cieśliński (2003a) cetraria islandica (l.) ach. soil also cieśliński (2003a) cladonia arbuscula (wallr.) flot. ssp. mitis (sandst.) ruoss soil also cieśliński (2003a) cladonia chlorophaea (flörke ex sommerf.) spreng. soil also cieśliński (2003a) cladonia coniocraea (flörke) spreng., nom cons. soil, trunk of pinus sylvestris, betula pendula, wooden crosses, decaying wood also cieśliński (2003a) cladonia fimbriata (l.) fr. soil, trunk of pinus sylvestris, wooden crosses, decaying wood also cieśliński (2003a) cladonia furcata (huds.) schrad. (ssp. furcata) soil also cieśliński (2003a) cladonia phyllophora hoffm. soil also cieśliński (2003a) cladonia rangiformis hoffm. soil also cieśliński (2003a) cladonia subulata (l.) weber ex f. h. wigg. soil also cieśliński (2003a) evernia prunastri (l.) ach. trunk of alnus glutinosa, acer platanoides, betula pendula, wooden crosses also cieśliński (2003a) flavoparmelia caperata (l.) hale trunk of alnus glutinosa cieśliński (2003a) hypocenomyce scalaris (ach.) m. choisy trunk of alnus glutinosa, pinus sylvestris, betula pendula, wooden crosses also cieśliński (2003a) hypogymnia physodes (l.) nyl. trunk of alnus glutinosa, pinus sylvestris, acer platanoides, betula pendula, quercus robur, juniperus communis, wooden crosses and bench, decaying wood also cieśliński (2003a) hypogymnia tubulosa (schaer.) hav. trunk of betula pendula imshaugia aleurites (ach.) s.l.f. meyer trunk of pinus sylvestris, wooden crosses, decaying wood also cieśliński (2003a) lecanora albescens (hoffm.) branth & rostr. stones, stone wall, concrete and stone tombstones lecanora carpinea (l.) vain. trunk of alnus glutinosa lecanora conizaeoides nyl. ex cromb. trunk of alnus glutinosa, pinus sylvestris, wooden crosses also cieśliński (2003a) lecanora dispersa (pers.) sommerf. stones, stone wall, concrete and stone tombstones lecanora polytropa (ehrh. ex hoffm.) rabenh. stone wall lichens of the holy hill 109 lecanora pulicaris (pers.) ach. trunk of alnus glutinosa also cieśliński (2003a) lecanora varia (hoffm.) ach. wooden crosses lecidella elaeochroma (ach.) m. choisy trunk of alnus glutinosa lecidella stigmatea (ach.) hertel & leuckert concrete tombstones lepraria sp. trunk of acer platanoides, alnus glutinosa, quercur robur, pinus sylvestris, juniperus communis, wooden crosses, decaying wood melanelixia fuliginosa (duby) o. blanco et al. ssp. glabratula (lamy) j. r. laundon trunk of alnus glutinosa, wooden crosses parmelia sulcata taylor trunk of alnus glutinosa, acer platanoides, quercus robur, betula pendula, wooden crosses and bench also cieśliński (2003a) parmelina tiliacea (hoffm.) hale wooden crosses parmeliopsis ambigua (wulfen) nyl. trunk of pinus sylvestris, wooden crosses also cieśliński (2003a) pertusaria amara (ach.) nyl. trunk of acer platanoides phaeophyscia nigricans (flörke) moberg concrete tombstones phaeophyscia orbicularis (neck.) moberg stones, stone tombstones phlyctis argena (spreng.) flot. trunk of acer platanoides, alnus glutinosa physcia adscendens h. olivier, nom. cons concrete tombstones physcia caesia (hoffm.) fürnr. stones, stone and concrete tombstones physcia dubia (hoffm.) lettau wooden crosses, stone wall placynthiella icmalea (ach.) coppins & p. james decaying wood also cieśliński (2003a) platismatia glauca (l.) w.l. culb. & c.f. culb. bark of betula pendula, wooden crosses pleurosticta acetabulum (neck.) elix & lumbsch in lumbsch, kothe & elix trunk of alnus glutinosa cieśliński (2003a) porpidia crustulata (ach.) hertel & knoph in hertel stone wall porpidia rugosa (taylor) coppins & fryday [=p. glaucophaea (körb.) hertel & knoph] erratic blocks cieśliński (2003a) protoparmeliopsis muralis (schreb.) m. choisy wooden crosses, stone and concrete tombstones also cieśliński (2003a) pseudevernia furfuracea (l.) zopf trunk of pinus sylvestris, juniperus communis, wooden crosses scoliciosporum chlorococcum (graewe ex stenh.) vězda trunk of alnus glutinosa, quercus robur, betula pendula, wooden crosses also cieśliński (2003a) trapeliopsis granulosa (hoffm.) lumbsch soil also cieśliński (2003a) tuckermanopsis chlorophylla (willd.) hale trunk of pinus sylvestris betula pendula, wooden crosses also cieśliński (2003a) usnea filipendula stirt. trunk of pinus sylvestris, betula pendula, wooden crosses usnea hirta (l.) f. h. wigg. trunk of pinus sylvestris, betula pendula, wooden crosses verrucaria muralis ach. concrete tombstones verrucaria nigrescens pers. concrete tombstones xanthoparmelia conspersa (ach.) hale stones also cieśliński (2003a) xanthoria polycarpa (hoffm.) th. fr. ex rieber trunk of juniperus communis, concrete tombstones, stone wall also cieśliński (2003a) tab. 1 cont. 110 a. matwiejuk erratic block in shaded and moist place. it is a very rare species, growing in very few stations on the polish lowland. epigeic lichens. the terricolous lichens grow in the holy hill vicinity in pine – tree forests. 10 species of terricolous lichens were identified on the soil, including 7 exclusive species (cetraria aculeata, c. islandica, cladonia arbuscula subsp. mitis, c. furcata, c. phyllophora, c. rangiformis). trapeliopsis granulosa grows on sandy road side – space. conclusions there is no data concerning the lichen biota of holy grabarka hill. several species of lichens from the forests surrounding the hill have been described by cieśliński (2003a). specific character of the holy hill, which is situated among forests has inclined to study its lichen biota, and varied stand of trees, wooden crosses, tombstones and stones have suggested that rare and interesting species of lichens can occur there. despite of a small area, differentiation of habitats and phorophits is so big that 64 species were identifid there. during our own studies the occurence of flavoparmelia caperata, pleurosticta acetabulum and porpidia rugosa, which were mentioned by cieśliński (2003a), was not confirmed. the lichens form a significant group of organisms in the scenery of the holy grabarka hill. their presence at all accessible substrata demonstrates positive results of synatropization. one of the distinctive features are wooden crosses overgrown abundantly by lichen thalus. acknowledgement. i wish to express my thanks to reviewer for his precious remarks and advice. referencens blanco o., crespo a., divakar p. k., esslinger t. l., hawksworth d. l., lumbsch h. t. 2004. melanelixia and melanohalea, two new genera segregated from melanelia (parmeliaceae) based on molecular and morphological data. mycol. res. 108 (8): 873–884. bystrek j. 1986. species of the genus bryoria brodo et hawksw. (lichenes, usneaceae) in europe. bulletin of the polish academy of sciences, biol. ser. 34 (10/12): 293–300. bystrek j. 1994. studien über die flechtengattungen usnea in europa.wyd. uniwersytetu marii curieskłodowskiej, lublin, 69 pp. cieśliński s. 2003a. distribution atlas of lichens (lichenes) in north-eastern poland. phytocoenosis 15 (n.s.), suppl. cartogr. geobot.15, 430 pp. cieśliński 2003b. red list of extinct and threatened lichenes in poland. (in:) k. cżyżewska (ed.). the threat to lichens in poland. monogr. bot. 91: 91–106. cieśliński s., czyżewska k., fabiszewski j. 2003. red list of exinct and threatened lichenes in poland. (in:) k. czyżewska (ed.). the threat to lichens in poland. monogr. bot. 91: 13–49. fałtynowicz w. 2003. the lichens lichenicolous and allied fungi of poland. an annotated checklist. szafer institute of botany, polish academy of sciences, kraków, 435 pp. kondracki j. 1994. geografia polski. mezoregiony fizyczno-geograficzne. pwn, warszawa. lechowski a. 2004. między niebem a ziemią. grabarka. góra krzyży (the hill of crosses). wydawnictwa orthdruk, białystok. radziukiewicz a. 2000. prawosławie w polsce. wydawnictwo arka, białystok. radziukiewicz a. 2001. góra krzyży i modlitwy. wydawnictwo orthdruk, białystok. dz. u. nr 168, poz. 1765, rozporządzenie ministra środowiska z dnia 9 lipca 2004 r. w sprawie gatunków dziko występujących grzybów objętych ochroną. lichens of the holy hill 111 porosty świętej góry, prawosławnego sanktuarium w grabarce s t r e s z c z e n i e w pracy przedstawiono wyniki badań nad porostami świętej góry, prawosławnego sanktuarium w grabarce. święta góra grabarka jest najważniejszym sanktuarium prawosławnym w polsce. położona jest w polsce północno-wschodniej, na podlasiu, między bugiem a narwią, na wysoczyźnie drohickiej, w południowej części niziny północnopodlaskiej. święta góra grabarka zasłynęła z cudu, jaki miał miejsce na początku 1710 roku podczas epidemii cholery, która dziesiątkowała ludność na terenie podlasia. święta góra grabarka nazywana jest górą krzyży. nazwa ta wywodzi się od krzyży ofiarnych z epitafiami pisanymi cyrylicą otaczającymi cerkiew przemienienia pańskiego. krzyże stojące wokół niewielkiej, drewnianej cerkwi stały się symbolem świętej góry. dziś na wzgórzu wznosi się około 10 tysięcy krzyży – drewnianych, metalowych, kamiennych, małych i wielkich. krzyże tradycyjnie przynoszone są przez wiernych od setek lat. pątnicy stawiają je obok sanktuarium przez cały rok, jednak najwięcej krzyży przybywa w przeddzień święta przemienienia pańskiego (19 sierpnia). za lasem krzyży znajduje się cmentarz oraz kompleks zabudowań klasztoru świętych marty i marii. żeński klasztor prawosławny powstał w 1947 roku, a w 1957 została wzniesiona cerkiew p. w. matki bożej wspomożycielki i opiekunki. na terenie świętej góry grabarki i jej okolic odnotowano 64 gatunki porostów (tab. 1), z 35 rodzajów. najliczniej reprezentowane są rodzaje cladonia – 8 gatunków, lecanora – 7, caloplaca – 4. porosty występują we wszystkich podstawowych formach morfologicznych. najliczniej reprezentowane są porosty epifityczne. 11 gatunków umieszczonych jest na czerwonej liście porostów wymarłych i zagrożonych w polsce (cieśliński et al. 2003), w tym 3 gatunki w kategorii wymierających – en (bryoria crispa, flavoparmelia caperata, pleurosticta acetabulum), 6 w kategorii narażonych – vu (cetraria islandica, parmelina tiliacea, porpidia rugosa, tuckermanopsis chlorophylla, usnea filipendula, u. hirta), 2 – w kategorii bliskich zagrożeniu – nt (evernia prunastri, hypogymnia tubulosa). 2014-01-01t11:47:35+0100 polish botanical society volvariella caesiotincta p. d. orton, a new species in the mycobiota of poland marek halama museum of natural history, wrocław university sienkiewicza 21, pl-50-335 wrocław, marhalam@biol.uni.wroc.pl halama m.: volvariella caesiotincta p. d. orton, a new species in the mycobiota of poland. acta mycol. 44 (1): 43–48, 2009. the article presents the first record of volvariella caesiotincta p.d. orton in poland. fruit body of the species was found on 22nd july 2002 in an oak-hornbeam/elm-ash municipal wood in the eastern part of wrocław. the saproxylic volvariella caesiotincta produced its carpophore on the base of rotten log of quercus robur. the article brings closer taxonomic profile, ecological requirements and distribution pattern of the species. it also describes macroscopic and microscopic characteristics of the discovered specimens and presents the specification of habitat the fungus concerned. key words: volvariella caesiotincta, macromycetes, urban greenery, wrocław, sw poland introduction the genus volvariella speg. includes fungi with pluteoid habit of the carpophores, forming distinctive saccate velum universale at the base of central stipe. the saccate velum forms distinct memebranous volva, that usually breaks up into lobes with time (boekhout 1990; skirgiełło 1999). the genus includes species which are saprotrophic, terrestrial, wood occupying, or living on decaying agarics, cosmopolitan, or nearly so fungi. there are c. 50 described volvariella species worldwide and 13 of them occuring in europe (kirk et al. 2008; horak 2005; singer 1986; krieglsteiner 2003). there have been reported 9 species of the genus from poland so far, i.e.: v. bombycina (schaeff.: fr.) singer, v. gloiocephala (dc.: fr.) boekhout & enderle, v. hypopithys (fr.) m. m. moser, v. media (schumach.: fr.) singer, v. murinella (quél.) m. m. moser, v. pusilla (pers.: fr.) quél., v. surrecta (knapp) singer, v. taylorii (berk. & broome) singer, and v. volvacea (bull.: fr.) singer (wojewoda 2003). seven of the species mentioned above are on the red list of the macrofungi in poland (wojewoda, ławrynowicz 2006). volvariella caesiotincta p.d. orton has never previously been acta mycologica vol. 44 (1): 43–48 2009 44 m. halama found in poland. however it should be noted, that after the author’s record, v. caesiotincta was reported at least from one other (not legitimately published) locality within polish area (puszcza niepołomicka forest; karasiński 2006). methods material was gathered within urban greenery of wrocław. the investigated habitat was illustrated with a phytosociological record. the floristic composition and diversity of the forest community were studied using the brown-blanquet method. the nomenclature of forest communities follows matuszkiewicz (2007); that of vascular plants follows mirek et al. (2002). microscopical slides of dried material were prepared with 5% nh4oh and congo red in 1% ammonia. the microcharacters of one recorded basidioma were observed and measured under a standard light microscope. morphological measurements were quoted according to breitenbach & kränzlin (1991). terminology of morphological and anatomical elements was adapted from vellinga (1988). size of spores, basidia, cheilocystidia and pleurocystidia, as well as pileipellis elements dimensions were correspondingly based on: 51, 31, 31, 21 and 21 measurements. drawings were made with the aid of a drawing tube under an oilimmersion objective. the collected specimen is deposited in the herbarium of the museum of natural history, wrocław university, wrocław (wrsl). results the occurrence of volvariella caesiotincta carpophore was discovered on 12th july 2002 in las wojnowski municipal forest. the forest area where the fungus occurred is surrounded by fields under cultivation and is situated in the eastern part of wrocław (fig. 1). a potential plant community there in the area is elm-ash community (ma-(ma-matuszkiewicz 2007). analysis of phytosociological relevés made within the area did not allow to determine an unambiguous association character of the forest community. representative plants of ficario-ulmetum minoris knapp 1942 em. j.mat. 1976 association have been left their stamp on floristic composition profile, but distinct participation of key species from carpinion betuli issl. 1931 em. oberd. 1953 alliance is evident too. the dominant species of the tree layer at the community where the fungus occurred are: carpinus betulus, fraxinus excelsior, quercus robur and tilia cordata. the shrub layer includes mainly: carpinus betulus, corylus avellana, sambucus nigra and padus avium. in the herb layer, ficaria verna, anemone nemorosa, milium effusum and maianthemum bifolium grow abundantly during spring season. in the course of field working only one specimen of volvariella caesiotincta was found. the carpophore of the fungus occupied the base part of distinct rotten log of quercus robur. further observations at the investigated site were conducted in july volvariella caesiotincta 45 2003 and july 2004, but no v. caesiotincta carpophores were observed. the location of the species has been carefully marked and will be monitored in the future. description of the specimens volvariella caesiotincta p.d. orton, bull. mens. soc. linn. lyon 43: 319. 1974. – pluteaceae, agaricales, agaricomycetidae, agaricomycetes, basidiomycota, dikarya, fungi (kirk et al. 2008; hibbett et al. 2007). macroscopic and microscopic characters. pileus 44 × 39 × 11.5 mm, applanate, with an obtuse umbo and slightly inflexed margin, fleshy, surface finely radially fibrillose at margin to nearly strigose at the central part, dry and mat, ± white, greish white, paler towards margin, with slight olivaceous tinge at the central part. lamellae crowded, free, thin, broadly ventricose, rounded at margin of pileus, flesh-pink, with whitish flocculose edge. stipe 52 × 5.0 mm, tapering upwards, with slightly bulbous base (up to c. 9.5 mm wide) enclosed by 3-lobed, greyish white to greyish brown, membranous volva (up to c. 21 mm high), solid, extremely fragile, glabrous to fibrillose-striate, translucent white. context in pileus and stipe quite compact, fragile, whitish (fig. 2). smell rather strong, reminding geranium robertianum odour. taste moderately strong, unpleasant, rather astringent. colour of spore print was not recorded. spores (6.2) 6.7 ± 0.3 (7.7) × (4.2) 4.6 ± 0.3 (5.4) µm, q = (1.21) 1.46 ± 0.08 (1.64), ellipsoid, oblong to oblong-ovoid, not ornamented. basidia 21-28 × 6.5-9.0 µm, clavate, with 4 sterigmata, without basal clamp. cheilocystidia 34-65 × 9.6-19 µm, ± clavate, frequently with apical papilla or up to 15 µm long, finger-like appendage, or lageniform to utriform. pleurocystidia 35-46 × 10-25 µm, clavate, lageniform to utriform, very rare. pileipellis: a cutis, made up of cylindrical elements (15-27 µm width), with intracellular, pale (yellow) pigment (fig. 3). observed septa without clamps. fig. 1. the location of the volvariella caesiotincta: a – site in wrocław; b – in poland (based on a 100 km atpol grid); 1 – urban boundary, 2 – municipal forests and parks, 3 – locality of the species. 46 m. halama material examined. wrocław (poland), las wojnowski forest, distinctly rotten log of quercus robur, 12.07.2002, coll. m. halama, wrsl. taxonomical remarks. nearly identical carpophores produces v. murinella, which can be separated with certainty from v. caesiotincta only on the base of microscopic characters. as it was emphasized by breitenbach et kränzlin (1995), in contrast to v. caesiotincta, v. murinella has cheilocystidia without the conspicuous finger-like and occasionally forked apical projection. moreover, there is an ecological difference between the two species, i.e. v. caesiotincta grows on wood or is associated with wood, while the latter occurs on humus rich soil or remains of leaves or plants and among grass. volvariella pusilla var. taylorii (berk. & broome) boekhout also resembles v. caesiotincta, from which it mainly differs in habitat (terrestrial fungus occurs mainly in grasslands on clayey soil), sweet and fungoid smell, indeterminate taste, and smaller size of carpophores (cf. boekhout 1990; skirgiełło 1999). habitat and general distribution. carpophores of volvariella caesiotincta almost exclusively appears in summer to early fall season (july-september), solitary or more rarely gregarious, on distinctly rotten wood (of optimal and terminal decay phase) of broadleaved trees, i.e. on logs and stumps of carpinus, fagus, tilia, ulmus, fraxinus and quercus (boekhout 1990; krieglsteiner 2003). volvariella caesiotincta is reported from north africa (marocco) and europe. in europe it is widely distributed and is known from spain, italy, france, liechtenstein, austria, hungary (krieglsteiner 2003), holland (boekhout 1990), great fig. 3. a – spores; b – basidia; c – cheilocystidia of volvariella caesiotincta recorded in wrocław (drawn by m. halama). volvariella caesiotincta 47 britain (b.m.s. 2004), switzerland (breitenbach, kränzlin 1995), czech republic (antonín et al. 1995), slovakia (lizoň 2001), germany (krieglsteiner 1991), denmark (vesterholt et al. 1998), sweden (gärdenfors 2005), norway (bendiksen et al. 1999) and finland (rassi et al. 2001). the fungus is recognized all over as a rare species (krieglsteiner 2003) and is included in the red lists of number of european countries. for example, in sweden it is included in indeterminate category of threat (dd) (gärdenfors 2005), in slovakia – in near threatened category (lr) (lizoň 2001), in denmark and norway – in vulnerable category (v) (vesterholt et al. 1998; bendiksen et al. 1999), in finland – is treated as critically endangered species (category cr) (rassi et al. 2001) and in germany it appears in group of very rare species (category r) (benkert at al. 1992). acknowledgment. financial support within a grant no. 6p04f01720 awarded by the state committee for scientific research is gratefully acknowledged. references antonín v., fellner r., herink j., lazebníček j., lizoň p., kotlaba f., šebek s. 1995. huby. 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(in:) z. mirek (ed.). biodiversity of poland. 1. w. szafer institute of botany, polish academy of sciences, kraków, 442 pp. rassi p., alanen a., kanerva t., mannerkoski i. (eds). 2001. suomen lajien uhanalaisuus 2000 (the 2000 red list of finnish species). ympäristöministeriö i suomen ympäristökeskus (ministry of the environment and finnish environment institute), helsinki, 432 pp. singer r. 1986. the agaricales in modern taxonomy. 4th ed. koeltz scientific books, koenigstein, 981 pp. skirgiełło a. 1999. flora polski. grzyby (mycota). 27: łuskowcowate (pluteaceae). w. szafer institute of botany, polish academy of sciences, kraków, 79 pp. vellinga e.c. 1988. glossary. (in:) c. bas, th. w. kuyper, m. e. noordeloos, e. c. vellinga (eds). flora agaricina neerlandica. critical monographs on families of agarics and boleti occurring in the netherlands. 1. a. balkema, rotterdam: 54–64. vesterholt j., lange ch., heilmann-clausen j., lassoe th., rald e. 1998. svampe. (in:) m. stoltze, s. pihl (eds). rodliste 1997 over planter og dyr i danmark. miljoog energiministeriet. danmarks miljo undersogelser og skovog naturstyrelsen: 30–54. wojewoda w. 2003. checklist of polish larger basidiomycetes. krytyczna lista wielkoowocnikowych grzybów podstawkowych polski. (in:) z. mirek (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków. wojewoda w., ławrynowicz m. 2006. red list of the macrofungi in poland. czerwona lista grzybów wielkoowocnikowych w polsce. (in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. w. szafer institute of botany, polish academy of sciences, kraków: 55–70. volvariella caesiotincta, nowy gatunek dla mikobioty polski streszczenie w artykule przedstawiono pierwsze w polsce stanowisko pochwiaka błękitnawego volvariella caesiotincta. dnia 22 lipca 2002 roku znaleziono owocnik v. caesiotincta na silnie zbutwiałej kłodzie quercus robur w zbiorowisku grądowo-łęgowym na terenie lasu lasu wojnowskiego we wrocławiu. w pracy przedstawiono charakterystykę cech makroskopowych i mikroskopowych zebranego okazu, zaprezentowano charakterystykę siedliskową odnotowanego stanowiska, a także przybliżono specyfikę taksonomiczną, ekologiczną i chorologiczną odnotowanego gatunku. fig. 2. fruitbody of volvariella caesiotincta recorded in wrocław (22. july 2002; photo and scans by m. halama). 2014-01-01t11:48:45+0100 polish botanical society new records of the annulate pluteus in european and asian russia ekaterina malysheva1, olga morozova1 and elena zvyagina2 1komarov botanical institute, 2 popov street, rus-197376, st. petersburg, ekama3@yandex.ru 2yugansky nature reserve, vil. ugut, rus-628458 surgut district, tumen region, helen.zvyagina@mail.ru m a l y s h e v a e. f., m o r o z o v a o. v., z v y a g i n a e. a.: new records of the annulate pluteus in european and asian russia. acta mycol. 42 (2):153-160, 2007. new records of the annulate pluteus were made by the authors in central russia (zhigulevsky nature reserve) and western siberia (yugansky nature reserve). the description of the species based on these records is presented. the taxonomic value of such features as the presence of velum and the color of lamellae edge as well as the similarity between chamaeota and pluteus are discussed and the new combination pluteus fenzlii is proposed. key words: chamaeota, pluteaceae, pluteus fenzlii, central russia, western siberia, nature reserves introduction for more than 100 years the generic name chamaeota (w.g. sm.) earle has been in use for designation the pluteus-like species with annulus. the genus chamaeota is one of the poorly investigated taxa of agaricoid fungi. at present the volume of the genus is uncertain. it totals 9 species according the index fungorum (http://www.indexfungorum.org), as soon as only two species on the data of the last issue of the dictionary of fungi (k i r k et al. 2001). the species distribution, ecology and morphology remain unknown because this genus was not studied on a global scale. there are some information about records of the species of chamaeota in europe, asia, northern and central america (s i n g e r 1978; y i n g 1995; m i n n i s et al. 2006). probably, the genus as a whole is cosmopolitan (s i n g e r 1978) and its species grow on all continents, except for antarctica, though the records are very rare. till now the representatives of chamaeota has been recorded on the territory of russia (the caucasus) only once by s i n g e r (1978). unfortunately, those specimens were not kept. the species of this genus inhabit usually large fallen trunks and rotten wood of deciduous trees. acta mycologica vol. 42 (2): 153-160 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 154 e. malysheva et al. for the first time chamaeota was separated as a subgenus from the large genus agaricus l.: fr. by w.g. smith in1870. f. earle has risen the taxon to the generic rank in 1909. the taxonomic position of chamaeota was changed repeatedly because of the reformation of the system of agaricales. it was initially considered (together with pluteus fr. and volvariella speg.) within agaricaceae chevall., then within amanitaceae r. heim ex pouzar, and after the segregation of pluteaceae kotl. et pouzar in 1972 up to the present within pluteaceae. the basic criteria used to distinguish the genus remain constant. the character of velum is recognized as the main of them. however the problem of relevance of the genus and its similarity to pluteus was repeatedly raised. so s i n g e r (1975) noted strong resemblance of chamaeota species known to him (ch. mammillata (longyear) murrill, ch. sphaerospora (peck) kauffm. and ch. fenzlii (schulzer) singer) to the representatives of pluteus (section hispidoderma fayod) by the micromorphological characteristics. in his earlier papers (s i n g e r 1958) he emphasized that character of velum, as well as its presence or absence, are not the sufficient criterion for delimitation of genera in many groups of agaricoid fungi. he gave as an instance the genus russula pers. that was remained stable despite of inclusion of several tropical species with the velum. besides, annulate p. atroavellaneus murrill was described from south america. singer has also described two south-american species of pluteus with rudimental volva (p. stephanobasis singer, p. circumscissus singer) in cited paper. it confirms his position in this problem. however, he refrained from any nomenclatural changes concerning the genus chamaeota because of scarcity of the available descriptions, absence of the type material, rarity of records. the taxonomic value of such feature as velum is considered in detail by g o r o v o j (1990). he has studied ontogenesis of many agaricoid taxa and has made a conclusion about low taxonomic weight of the features of the basidiocarp development and covering structures. these features are subject substantially to parallel variability in many related and phylogenetic remote groups. moreover detailed study of ontogenesis of pluteus species (r e i j n d e r s 1963; g o r o v o j 1990) has shown that pilangiocarpous and stipitangiocarpous types of basidiocarp development are attributes of several species (p. admirabilis peck, p. chrysophlebius (berk. et ravenel) sacc). the new data basing on the molecular research are the most serious reason for the instability of the genus chamaeota concept. according to the recent paper devoted to the study of the american species with application of the molecular methods (m i n n i s et al. 2006), chamaeota does not form an independent clad, as against volvariella, but is nested inside pluteus clade. species ch. mammilata studied occupies a very close position to p. ephebeus (fr.: fr.) gillet on cladogram. moreover both species are very similar to each other in their micromorphological characteristics. therefore new combination p. mammilatus (longyear) minnis, sundb. et methven seems reasonable to us. the further morphological studies of chamaeota species (especially of those with cellular pileipellis) supported by the molecular data, could clear their taxonomic position as well as volume of the genus. the given morphological study of the european species ch. fenzlii very closed to p. mammilatus also has permitted to find out the features of its similarity with the representatives of the genus pluteus and to classify it as the pluteus. new records of the annulate pluteus 155 materials and methods the study was based on the material collected by the authors in 2000-2006 in the territory of two nature reserves: the zhigulevsky (3 collections) and the yugansky (4 collections). the zhigulevsky nature reserve occupies the central part of the zhiguli a broken highland in east of privolzhskaya plateau, on the right bank of volga river (samara region). the zhiguli consists of exposed carbonate rocks covered by loam layer in lower part. the climate is continental with annual precipitation 500 mm. the natural vegetation over the hills is represented by open pine forests (pinus sylvestris l.) in combination with steppes, in ravines by broad-leaved forests of tilietonemoreta type (k l e o p o v 1990). in stands tilia cordata mill., acer platanoides l., quercus robur l., ulmus glabra huds., corylus avellana l. are predominate. the yugansky nature reserve is located in the basin of bolshoy yugan river (tumen region). the geological structure is formed by the quaternary deposits, mainly by the pleistocene ones covered with loamy and clay soils. the climate of this area is moderately continental. the annual precipitation totals 650 mm. the main types of vegetation are coniferous forests (pinus sibirica du tour, abies sibirica ledeb., picea obovata ledeb.) and secondary deciduous-coniferous forests (with populus tremula l. and betula pendula roth) with mossy or grassy cover. the different types of swamps occupy 20-30% of the territory. the collections were made, documented and preserved with standard methods (b o n d a r t s e v , s i n g e r 1955). macroscopic description is based on the study of the material in fresh and dried condition as well as the analysis of the photos. the dried material was examined using standard microscopic techniques. spores, basidia and cystidia were observed in squash preparations of small parts of the lamellae in 5% koh. the pileipellis was examined in the preparation of the radial section of the pileus. microscopic measurements and drawings were made at 600x with micmed 2-2 microscope. basidiospore dimensions are based on observation of 30 spores per each of 8 collections, cystidia dimensions on observation of at least 10 structures per collection. spore length to width ratios are reported as q. mean values for q are designated as q∗. the collected material is deposited in mycological herbarium of the komarov botanical institute (le). results and discussion below we propose a new combination. the description of the species is based on the study of specimens collected by the authors in the territory of central russia and western siberia. pluteus fenzlii (schulzer) e. malysheva, morozova et zvyagina comb. nov. basionym: agaricus fenzlii schulzer, verh. zool.-bot. ges. wien 16: 49. 1866. synonyms: annularia fenzlii (schulzer) gillet, 1876; chamaeota fenzlii (schulzer) singer, 1978. pileus 17-70 mm in diam., initially obtuse-conical to become campanulate-convex, convex and flattened, usually with broad umbo, with entire, involute at first margin, sometimes cracked when old, not hygrophanous, translucently striate at the 156 e. malysheva et al. margin only, dry, vividly yellow or with orange tinge, slightly darker at centre, radially fibrillose sometimes becoming rimose at margin (with visible white background), covered by distinct yellow to brownish appressed squamules or hairs, erected over the center. appearance of pileus looks like ones of volvariella species (figs 1, 2). lamellae free, crowded to subdistant (approximately 12 on centimeter near margin), with lamellulae, thin, ventricose, ap to 5 mm broad, pale pink to grayish pink, with entire edge which can be concolorous, white or distinctly yellow (yellow hue can disappear with age) (fig. 3). stipe 25-50 × 4-10 mm, central to slightly excentric, cylindric, slightly broadened towards base but without basal bulb, smooth, whitish to pale yellow above a ring zone, with longitudinal yellow to brown-yellow fibrils in the lower part, with white basal tomentum. annulus entire, sheathing, fluffy or flake like, but often fragmentary and evanescent, remains as a ring zone, white to yellowish white, disposed on central or lower part of stipe. flesh of pileus and stipe solid, white, slightly yellow under pileus cuticle. odor and taste not distinctive. spores 4.2-7.6 × 4.0-6.5 μm, q = 1.00-1.33 (q∗ = 1.17), broad-ellipsoid to subglobose to slightly ovate in profile and face views, with a small hilar appendix and single large central oil drop, sometimes with granular contents (when oil drops are numerous), smooth, thin-walled to slightly thick-walled, hyaline to pale yellow in koh, non-amyloid. basidia 4-spored, 16-25 × 6.5-10 μm, clavate, often with taper-μm, clavate, often with taper-m, clavate, often with tapering apex, hyaline, thin-walled. cheilocystidia 22-73 × 8-31 μm, very abundant, often aggregated to continuous layer, variable in appearance, clavate, narrow-fusoid to clavate-ventricose, fusoid-ventricose and broad-lageniform, with short or slightly lengthened necks, narrow at base, hyaline in koh, thin-walled or thick-walled, usually with granular yellow contents and drops on apex. pleurocystidia 32-81 × 10-32 μm, abundant, mainly lageniform, rarely fusoid-ventricose, strongly inflated, with short or long (to 20 μm length and 8 μm wide) necks, narrow at base, hyaline in koh, thin-walled, with oil drops on apex. lamellae trama inverse. pileipellis is plagiotrichoderm consisting of periclinal basal cylindrical hyphae (5.5-8 μm wide) with obliquely to fully erect hyphal ends. basal hyphae thin-walled, containing yellowbrown intracellular or slightly incrusting pigment. superficial hyphal ends consist of chains of short or lengthened inflated (up to 12 μm) cells, thin-walled, with brown intracellular pigment. these hyphae dispose in bundles forming squamules (fig. 4). stipitipellis a cutis consisting of cylindrical hyphae, parallel to a surface, thin-walled, 5.5-8 μm wide, with or without brown intracellular pigment. �lamp connections ab-μm wide, with or without brown intracellular pigment. �lamp connections ab-m wide, with or without brown intracellular pigment. clamp connections absent. habitat on wood of deciduous trees, particularly on tilia, acer and betula, solitary or in small groups. specimens examined: central russia, samara region, zhigulevsky state nature reserve: vicinities of bakhilova polyana village, in tilia cordata-acer platanoides forest, on fallen trunk of tilia, 27 viii 2000, coll. and det. e. f. malysheva (le 246082). – vicinities of shiryaevo village, shiryaevskaya valley, in tilia cordataacer platanoides forest, on fallen trunk of deciduous tree, 17 viii 2004, coll. and det. e. f. malysheva (le 246085). – vicinities of bakhilova polyana village, in tilia cordata-acer platanoides forest, on fallen trunk of deciduous tree, 18 viii 2004, coll. o. v. morozova, det. e. f. malysheva (le 246083) (fig. 2). siberia, tumen region, surgut district, yugansky nature reserve: bank of lake, in swamp betula pendula forest, on fallen trunk, 18 vii 2006, coll. e. a. zvyagina, det. e. f. malysheva new records of the annulate pluteus 157 fig. 4. pluteus fenzlii (le 246083): a – pileipellis, b – basidiospores, c – basidium, d – cheilocystidia, e – pleurocystidia; scale bars a-� = 10 μm, d-e = 15 μm. 158 e. malysheva et al. (le 246084) (fig. 1). – bank of entl’turyah river, in populus tremula forest, on fallen trunk of betula, 16 viii 2006, coll. and det. e. a. zvyagina (le 235471). – basin of entl’turyah river, in populus tremula-betula pendula forest (with picea obovata and abies sibirica), on fallen trunk of betula, 17 viii 2006, coll. and det. e. a. zvyagina (le 235470). – bank of kolkochenyagun river, in swamp betula pendula forest, on fallen trunk of betula, 18 vii 2006, coll. and det. e. a. zvyagina (le 235469). pluteus fenzlii is similar to the american annulate species p. mammilatus in its habitus, pileus color, type of pileipellis and form of cheilocystidia. it differs from the latter only by presence of yellow lamellae edge. the similarity of these species has been mentioned by s i n g e r (1975). one more species – pluteus leoninus (schaeff.: fr.) p. kumm. – has some morphological resemblance to p. fenzlii. it differs by absence of annulus, by less squamulouse pileus and by possessing of pleurocystidia with excrescences. in the case of p. fenzlii with the reduced annulus these species can be confused in the field. all specimens of p. fenzlii examined in the present work showed considerable variation of most important diagnostic features, even within a single specimen (tab. 1). so, the annulus can be fugacious, fibrillose, presenting in the mature basidiocarp only as a poor visible ring zone, or well differentiated, almost membranous, at first entire, later breaking into patches. the yellow color of the edge of lamellae can be differently distinct – from almost invisible (located only in the edge of pileus or fig. 5. pluteus fenzlii (le 246083): a – cheilocystidiogram, b – pleurocystidiogram; scale bar = 10 μm. new records of the annulate pluteus 159 t ab le 1 t he v ar ia ti on o f t he m ac ro a nd m ic ro sc op ic fe at ur es in d if fe re nt p lu te us fe nz lii s pe ci m en s sp ec im en s sp or es a nn ul us c ol or o f l am el la e ed ge c he ilo -c ys ti di a p le ur oc ys ti di a le ng th m in –m ax (m ea n) w id th m in –m ax (m ea n) q q * l e 2 46 08 3 4. 8– 7. 6 (6 .4 ) 4. 6– 6. 5 (5 .5 ) 1. 00 – 1. 31 1. 17 fib ri llo se ye llo w 22 –6 0 × 8– 27 32 –6 7 × 12 –2 7 l e 2 46 08 5 5. 1– 7. 0 (6 .2 ) 4. 8– 6. 2 (5 .5 ) 1. 00 – 1. 29 1. 13 ri ng z on e co nc ol or ou s w it h si de ( in it ia lly p al e ye llo w ) 32 –5 4 × 12 –2 0 55 –6 2 × 13 .5 –2 1 l e 2 46 08 2 5. 5– 7. 8 (6 .5 ) 4. 4– 5. 5 (5 .0 ) 1. 06 – 1. 49 1. 28 ri ng z on e ye llo w 38 –7 3 × 8– 31 50 –8 0 × 16 –2 0 l e 2 46 08 4 4. 2– 7. 6 (6 .5 ) 4. 0– 6. 2 (5 .6 ) 1. 00 – 1. 30 1. 16 w el l d ev el op ed , al m os t m em br an ou s w hi te , p in k, ye llo w is h ne ar o f pi le us m ar gi n 27 –6 5 × 12 –2 2 40 –8 1 × 10 –3 2 l e 2 35 47 1 4. 6– 7. 4 (5 .9 ) 4. 0– 6. 0 (5 .1 ) 1. 00 – 1. 42 1. 16 fib ri llo se , fu ga ci ou s ye llo w 31 –1 15 × 9 –4 1 43 –6 7 × 10 –2 0 l e 2 35 47 0 4. 8– 6. 8 (6 .0 ) 4. 4– 6. 7 (5 .2 ) 1. 00 – 1. 35 1. 15 fib ri llo se , fu ga ci ou s w hi te , p in k 38 –7 7 × 9– 27 25 –5 2 × 7– 23 l e 2 35 46 9 4. 6– 7. 0 (5 .7 ) 4. 0– 5. 6 (5 .0 ) 1. 00 – 1. 42 1. 16 fib ri llo se , fu ga ci ou s co nc ol or ou s w it h si de ( in it ia lly p al e ye llo w ) 44 –5 7 × 8– 16 47 –6 2 × 10 –2 1 e xp la na ti on s: m ic ro sc op ic d im en si on o f ba si di os po re s ar e ba se d on o bs er va ti on o f 30 s po re s. a ll ot he r di m en si on s ar e ba se d on o bs er va ti on o f 10 s tr uc tu re s. q – le ng th /w id th ; q ∗ – m ea n m ea ni ng o f q . 160 e. malysheva et al. in a few lamellae) to entirely clear and vivid, especially in the young basidiocarps. the micromorphological characteristics are varying too. the size of spores, the form and the size of cheiloand pleurocystidia strongly variate in the single basidiocarp (fig. 5). a range of intermediate states is characteristic for analyzed features as well as there isn’t any correlation between these states. polymorphism of cystidia size and lamellae pigmentation shows that these features have little value by themselves for delimitation of the species. so, the low taxonomic value of the lamellae color for pluteus species has been demonstrated by vellinga (ve l l i n g a 1990), who considered p. luteomarginatus rolland with the yellow-marginate lamellae as a synonym of p. leoninus. taking into account all of this, it can be supposed that p. fenzlii and p. mammilatus are the same species. however, the populations growing in the different continents can possess some genetic differences. the additional genetic studies could answer to the question about similarity of the american and the european species of annulate pluteus. acknowlegments: the authors would like to thank dr. eugene popov for critical review and valuable comments and to dr. alexander kovalenko for kind permission to use the photo and for helpful discussion. this study was supported in part by russian foundation for basic research (project № 07-04-01408-а). references b o n d a r t s e v a. s., s i n g e r r. 1950. rukovodstvo po sboru vysshikh bazidialnykh grybov dlya ikh nauchnogo izuchenia (a guide to collecting of high basidial fungi for scientific study). proceedings of komarov botanical institute. 2 (6): 499–572. g o r o v o j l. f. 1990. morfogenez plastinchatykh gribov (morphogenesis of the gilled fungi). naukova dumka, kiev. 168 pp. k i r k p. m., c a n n o n p. f., d a v i d j. c., s t a l p e r s j. a. 2001. ainsworth & bisby’s dictionary of the fungi. ninth edition. cab international. 655 pp. k l e o p o v yu. d. 1990. analiz flory shirokolistvennykh lesov evropejskoj chasti sssr (the analysis of the flora of the broad-leaved forests of the european part of ussr). naukova dumka, kiev. 352 pp. m i n n i s a. m., s u n d b e r g w. j., m e t h v e n a. s., s i p e s s. d., n i c k r e n t d. l. 2006. annulate pluteus species, a study on the genus chamaeota in the united states. mycotaxon 96: 31–39. r e i j n d e r s a.f.m. 1963. les problémes du dévelopment des carpophores des agaricales et de quelques groupes viosins. junk, haag. 412 pp. s i n g e r r. 1958. contribution towards a monograph of the genus pluteus, especially those of the east slope of the andes and brazil. lloydia 21: 195–299. s i n g e r r. 1975. the agaricales in modern taxonomy. j. cramer, vaduz. 912 pp. s i n g e r r. 1978. keys for the identification of the species of agaricales ii. sydowia 31: 193–237. ve l l i n g a e. c. 1990. pluteaceae. (in:) c. b a s , th. w. k u y p e r , m. e. n o o r d e l o o s , e. c. ve l l i n g a . a. a. b a l k e m a (eds). flora agaricina neerlandica. critical monographs on families of agarics and boleti occurring in the netherlands. 2. pleurotaceae, pluteaceae, tricholomataceae. rotterdam. 137 pp. y i n g j.-zh. 1995. new and noteworthy agarics from china. i. new species of chamaeota. mycotaxon 54: 303–307. 2014-01-01t11:45:53+0100 polish botanical society fungi isolated in school buildings elżbieta ejdys department of mycology, university of warmia and mazury in olsztyn oczapowskiego 1a, pl-10-719 olsztyn-kortowo, elzbieta.ejdys@uwm.edu.pl e j d y s e.: fungi isolated in school buildings. acta mycol. 42 (2):245-254, 2007. the aim of the study was to determine the species composition of fungi occurring on wall surfaces and in the air in school buildings. fungi isolated from the air using the sedimentation method and from the walls using the surface swab technique constituted the study material. types of finish materials on wall surfaces were identified and used in the analysis. samples were collected in selected areas in two schools: classrooms, corridors, men’s toilets and women’s toilets, cloakrooms, sports changing rooms and shower. examinations were conducted in may 2005 after the heating season was over. fungi were incubated on czapek-dox medium at three parallel temperatures: 25, 37 and 40°c, for at least three weeks. a total of 379 isolates of fungi belonging to 32 genera of moulds, yeasts and yeast-like fungi were obtained from 321 samples in the school environment. the following genera were isolated most frequently: aspergillus, penicillium and cladosporoium. of the 72 determined species, cladosporium herbarum, aspergillus fumigatus and penicillium chrysogenum occurred most frequently in the school buildings. wall surfaces were characterised by an increased prevalence of mycobiota in comparison with the air in the buildings, with a slightly greater species diversity. a certain species specificity for rough and smooth wall surfaces was demonstrated. fungi of the genera cladosporium and emericella with large spores adhered better to smooth surfaces while those of the genus aspergillus with smaller conidia adhered better to rough surfaces. the application of three incubation temperatures helped provide a fuller picture of the mycobiota in the school environment. key words: fungi, school, walls, indoor air introduction schools belong to a specific category of public buildings were factors that increase the expansion of fungi belonging to different systematic and ecological groups accumulate. the technical condition of a school building which results from the construction technology, finish materials and its use significantly influences the occurrence of conditions favourable for a temporary or permanent occurrence of fungi (b o g a c k a 1997). school occupants constitute further sources or transmission vectors of fungi acta mycologica vol. 42 (2): 245-254 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 246 e. ejdys into and within a school building. the present author’s previous studies show that fungi are isolated in over 1/3 of school children (e j d y s 2003). the bioaerosol in school buildings also consists of secondary metabolites produced by the mycobiota occurring in building partitions. possible toxins penetrating the bodies of the children and school employees may cause various toxicoses (b u d a k 1998). on the other hand, antibiotics produced in low concentrations may induce resistance of the bacterial flora of school occupants (m a r k i e w i c z , k w i a t k o w s k i 2001). as literature data on the diversity range of fungi in schools are greatly divergent (l e v e t i n at al. 1995; m e k l i n at al. 2002; d a c a r r o et al. 2003), the species composition of fungi occurring on wall surfaces and in the air in school buildings was analysed. material and methods fungi isolated from the air using the sedimentation method and from the walls using the surface swab technique were the study material. types of finish materials on wall surfaces were identified and used in the analysis. surfaces finished with acrylic or emulsion paint were defined as rough while the walls covered with gloss paint or tiled were defined as smooth. macroscopic changes indicating the development of the mycelium were not found on the walls. samples were collected in selected areas in two schools: classrooms, corridors, men’s toilets and women’s toilets, cloakrooms, sports changing rooms and shower (only school i). a total of 312 samples were collected in 13 enclosed spaces within the school buildings. studies were conducted in may 2005 after the heating season was over. indoor temperature and air humidity were measured during sampling. fungi were incubated on czapek-dox medium at three parallel temperatures: 25, 37 and 40°c, for at least three weeks. cultured colonies of moulds were described macroscopically. slides stained with cotton blue in lactophenol according to g e r l a c h (1972) were made to identify microscopic features. biochemical properties were assessed using methods commonly accepted in mycological laboratories. the obtained isolates of yeast-like fungi and yeasts were incubated on sabouraud medium without antibiotics. media containing antibiotics that could affect typical fungal features were not used (d y n o w s k a 1991/1992). the isolates were incubated at 37°c for 48 hours. after fungal growth was obtained, the material was passed twice or three times onto fresh sabouraud medium to eliminate bacteria which relatively often accompanied fungi in the first culture. after pure bacteria-free strains were obtained, microcultures on nickerson agar were established. fungi were inoculated on glass slides, covered with a thin layer of the medium (ca. 2 mm), and 2-3 drops of broth and serum in a ration of 1:1 were added onto the inoculation site. microcultures were incubated at 37°c for 48 up to 72 hours. macroscopic features (size, colour, shape, texture, smell of the colony) and microscopic features (shape and size of budding cells, blastospores and chlamydospores, the diameter of pseudohyphae and hyphae) as well as biochemical features obtained on bio–mérieux api-tests (api 20c, api 20c aux) were used for identification. chromagar (bio–mérieux) was used to differentiate individual species of the fungi in school buildings 247 genus candida. it was treated as an auxiliary test as it does not yield accurate results in mixed-species isolations (b o u c h a r a et al. 1996). photographic documentation was made throughout the process. yeast-like fungi and yeasts were determined using the following keys: k r e g e r v a n r i j (1984); b a r n e t t , p a y n and ya r r o w (1990), and a study by -k u r n a t o w s k a (1995). fungi of the genera aspergillus and penicillium were identified up to the level of species using the following keys: r a p e r and f e n n e l l (1965) and r a p e r , t h o m and f e n n e l l (1945). other species were determined using the atlas of clinical fungi (d e h o o g et al. 2000). results a total of 379 fungal isolates were obtained from 312 samples. 213 isolates were recorded in 7 study areas in school i and 166 isolates in school ii. 32 genera of moulds, yeasts and yeast-like fungi were identified in the study material. fungi of the genera aspergillus, penicillium and cladosporium were isolated most frequently. fungi belonging to 27 and 21 genera were recorded in school i and ii, respectively; 16 genera occurred in the study areas in both buildings while the others were isolated only from one school and were usually sporadic isolates. 9 fungi of the genus chaetomium and 4 of the genus hormographiella were exceptionally recorded in school i (fig. 1). of the 72 fungal species identified in the samples, 33 species were isolated at least twice. a group of predominant species with a similar occurrence frequency can be distinguished. these are: cladosporium herbarum (43 isolates– 11.3%), aspergillus fumigatus (39-10.3%) and p. chrysogenum (37-9.8%) as well as p. citrinum (32-8.4%) (tab. 1). p. chrysogenum (13.1%), p. citrinum (11.7%) and cl. herbarum (10.3%) were isolated most frequently in school i. a. fumigatus occurred almost twice less frequently (6.6%). it was, however, one of the species predominant in the mycobiota in school ii, and its frequency of occurrence was 15.1%. cl. herbarum, whose prevalence was 12.7%, was the second dominant. p. chrysogenum and p. citrinum, on the other hand, occurred with the frequency lower than 4.5%. the differences in the species range in both buildings could also be noticed in the greater number of species belonging to the genus aspergillus recorded in building i (18) than in building ii (6). fig.1. occurrence of fungal genera in the school buildings. 248 e. ejdys 235 fungal isolates were obtained from the swabs and 144 isolates were obtained using the sedimentation method. 26 and 22 genera were recorded on wall surfaces and in the air, respectively; 16 genera were isolated in both sample types (fig. 2). only 6 genera were characteristic of the air. these were: bipolaris, bjerkandera, chrysosporium, corynespora, madurella and phoma. fungi of the genera penicillium (32 isolates), cladosporium (26) and aspergillus (22) were recorded most frequently in this medium. the same genera also dominated on wall surfaces although their stratification was slightly different. the genus aspergillus unquestionably dominated (75 isolates). yeast-like fungi, mostly of the genera candida, rhodotorula and geotrichum, were frequently recorded in both sample types: 11.3%. significantly greater differences were recorded in the species range in the wall samples and the air samples. twice as many species (65) were identified on the walls as in the air (32). however, the structure of the domination in both sample types was the same. 23 species common for both the walls and the air were recorded. of the 32 species that occurred only on the surfaces in the school buildings, 16 species represented the genus aspergillus. only one isolate of the genus candida, c. datilla, was determined from the air. the prevalence of yeast-like fungi in the air did not differ significantly from that on the wall surfaces and was 10.4% and 11.9%, respectively. fungi of the order mucorales which occurred sporadically in the school buildings did not occur in the air at all. a slightly smaller number of isolates was recorded on smooth walls (106) than on rough walls (129). rough surfaces were characterised by a slightly greater species range (43 species) and a greater number of fungi belonging to the genus aspergillus (15) than smooth walls (39 and 12 species, respectively). these differences were accompanied by an increased number of isolates of fungi of this genus: 45 on rough walls and 30 on smooth walls. the fungus emericella quadrilineata was identified on the smooth wall in the sports shower, and its asexual stage, aspergillus quadrilineatus, on the rough surface. however, a greater number of cladosporium species were recorded on smooth surfaces. apart from cl. herbarum, cl. cladosporoides, cl. oxysporum and cl. sphaerospermum also occurred. significant differences in the species range and prevalence of fungi belonging to the genus penicillium on both surfaces studied were not recorded. fig. 2. occurrence of fungal genera on the walls and in the air in the school buildings. fungi in school buildings 249 ta b l e 1 fungi isolated from the walls and the air in the school buildings no fungal species number of isolates to ta l ro ug h w al l sm oo th w al l ai r sc ho ol i sc ho ol i i 1 acremonium alabamense morgan-jones 1 1 0 0 1 0 2 ac. hyalinulum (sacc.) w. gams 1 0 1 0 1 0 3 ac. kiliense grütz 1 0 1 0 0 1 4 ac. roseogriseum (s.b. saksena) w. gams 1 0 0 1 1 0 5 acremonium sp. 1 0 0 1 0 1 6 alternaria alternata (fr.) keissl. 2 1 0 1 0 2 7 a. infectoria simmons 1 0 1 0 1 0 8 alternaria sp. 2 2 0 0 0 2 9 artrographis kalrae (tewari&macpherson) singler & carmichael 1 0 1 0 1 0 10 aspergillus allahabadi mehrotra & agnihotri 1 1 0 0 0 1 11 a. aureolus fennell & raper 1 0 1 0 1 0 12 a. auricomus (gueguen) saito 1 0 1 0 1 0 13 a. caesiellus saito 5 3 2 0 1 4 14 a. candidus link 3 1 1 1 1 2 15 a. crystallinus kwon & fennel 1 1 0 0 1 0 16 a. flavus link 12 7 5 0 7 5 17 a. fumigatus fresenius 39 13 8 18 14 25 18 a. glaucus link 1 1 0 0 1 0 19 a. granulosus raper & thom 1 1 0 0 1 0 20 a. janus raper & thom 4 3 1 0 3 1 21 a. longobasidia (bain.) moseray 1 0 1 0 1 0 22 a. malodoratus kwon & fennell 1 1 0 0 1 0 23 a. nidulans (eidam) wint 1 0 1 0 1 0 24 a. niger van tieghem 4 2 1 1 4 0 25 a. ochraceus wilhelm 1 1 0 0 1 0 26 a. quadrilineatus thom & raper 1 1 0 0 1 0 27 a. sydowi (bain.& sart.) thom & church 3 0 3 0 3 0 28 a. versicolor tiraboschi 12 6 4 2 6 6 29 aspergillus sp. 4 3 1 0 1 3 30 bipolaris spicifera (bain.) subram. 2 0 0 2 2 0 31 bjerkandera sp. 4 0 0 4 2 2 32 botrytis cinerea pers. ex pers. 2 0 2 0 0 2 33 candida albicans (robin) berkhout 1 1 0 0 0 1 34 c. datila (kluyver) s.a. meyer & yarrow 1 0 0 1 0 1 35 c. globosa yarrow & s.a. meyer 5 0 5 0 2 3 36 c. guilliermondii (castellani) berkhout 1 1 0 0 0 1 37 c. krusei (castellani) berkhout 1 1 0 0 0 1 38 c. versatilis (etchells & t.a. bell) s.a.meyer & yarrow 2 0 2 0 2 0 250 e. ejdys 39 chaetomium atrobruneum ames 9 3 5 1 9 0 40 chrysosporium queenslandicum apinis & rees 1 0 0 1 1 0 41 cladosporium cladosporoides (fres.) de vries 3 2 0 1 3 0 42 cl. herbarum (pers.) link: fr. 43 13 10 20 22 21 43 cl. oxysporum berk. & curt. 3 1 0 2 0 3 44 cl. sphaerospermum penz. 4 1 0 3 1 3 45 corynespora cassiicola (berk. & curt.) wei 1 0 0 1 0 1 46 emericella quadrilineata (thom & raper) c.r. benjamin 1 0 1 0 1 0 47 emonsia crescens emmons & jellison 1 1 0 0 1 0 48 epidermophyton floccosum (harz) langer. & milochevitch 3 1 1 1 1 2 49 fusarium incarnatum (rob.) sacc. 3 1 1 1 1 2 50 geotrichum candidum link: fr. 9 2 4 3 3 6 51 g. clavatum de hoog et al. 4 0 1 3 4 0 52 gymnoascus dancaliensis (castell.) v. arx 2 1 0 1 2 0 53 hormographiella aspregillata guarro et al. 2 1 0 1 2 0 54 h. verticillata guarro et al. 2 2 0 0 2 0 55 madurella grisea mackinnon et al. 1 0 0 1 0 1 56 microsporum fulvum uriburu 1 1 0 0 1 0 57 mucor amphibiorum schipper 1 0 1 0 0 1 58 mucor ramosissimus samutsevich 1 0 1 0 1 0 59 penicillium camamberti thom 1 0 1 0 0 1 60 p. chrysogenum thom 37 12 10 15 28 9 61 p. citrinum thom 32 11 8 13 25 7 62 p. claviforme bainier 1 0 1 0 0 1 63 p. notatum westling 6 3 1 2 3 3 64 p. purpurrescens (scopp) n. comb. 1 1 0 0 1 0 65 p. steckii zaleski 4 0 2 2 0 4 66 phialophora pubakii (laxa) schol-schwarz 1 1 0 0 1 0 67 phoma cruris-hominis punithalingam 1 0 0 1 1 0 68 rhizopus sp. 1 1 0 0 0 1 69 rhodotorula glutinis (fresenius) f.c. harrison 12 3 2 7 8 4 70 rhodotorula sp. 5 3 2 7 4 1 71 scopulariopsis brevicaulis (sacc.) bain. 1 1 0 0 0 1 72 s. brumptii salvanet-duval 3 0 1 2 1 2 73 s. flava (sopp) morton & g. smith 2 1 1 0 2 0 74 scopulariopsis sp. 1 1 0 0 1 0 75 saccharomyces cerevisiae meyen ex e.c. hansen 5 2 1 2 2 3 76 saccharomycopsis capsularis schiönning 2 2 0 0 1 1 77 scytalidum lignicola pesante 1 0 1 0 0 1 78 trichphyton tonsurans malmsten 1 0 0 1 1 0 79 t.verrucosum bodin 1 0 0 1 0 1 80 trichophyton sp. 4 0 4 0 4 0 81 unidentified isolates 34 5 4 25 12 22 total 379 129 106 144 213 166 tab. 1 cont. fungi in school buildings 251 humidity ranged between 43 and 49% in school i and between 48 and 52% in school ii. these values correspond to the pn-iso 8756: 2000 standard on the air quality. the temperature was quite low in school ii and ranged from 17.0 to 18.6 c°. it was warmer in school i, where the temperature ranged from 19.8 to 23.5 c°. discussion the composition of indoor mycobiota may differ significantly due to the nature and physicochemical properties of specific indoor environments, their use and the number of occupants, also not excluding the time of exposure to fungi. results of most mycological analyses show that so-called indoor fungi of the genera aspergillus and penicillium as well outdoor fungi, or “field fungi”, such as alternaria or cladosporium, are recorded most frequently inside buildings (l e v e t i n at al. 1995; g u t a r o w s k a et al. 2004). in his analysis of the species range of indoor air in 19 groups of building facilities used for different purposes, z y s k a (2001) demonstrated 14 recurring species of the 227 identified fungi. in the present study, the occurrence of 7 species was confirmed: alternaria alternata, aspergillus flavus, a. fumigatus, a.niger, a.versicolor, cladosporium herbarum, penicillium chrysogenum. they include species dominant in the school buildings studied. the range of fungal species that characterises indoor environment constitutes contamination of the pool of indoor fungi and species diffused with the air from outdoor environment (b o g a c k a 1997). it is best visible in spring when after a long heating season increased ventilation begins without the need to prevent heat losses. indoor temperature similar to that outdoors may also cause temporary enrichment of the mycobiota in buildings although there are usually fewer spores in the air in spring than in summer or autumn (s h e l t o n et al. 2002; to p b a s et al. 2006). thus, the great number of species isolated in the present study may indicate temporary contamination of the buildings by fungi from outside while those isolated only in the air may indicate incoming species. the greater taxonomic differentiation of the mycobiota in school i may result from its location in a park, with many trees and shrubs, some fountains, incorrectly performed drainage and, consequently, prolonged stagnation of rain waters. the latter may be of great importance in the context of the increase in air humidity which is one of the basic factors limiting the occurrence of moulds. the age of the buildings is also an important factor encouraging contamination by fungi. both schools are less than 30 years old and were constructed with prefabricated concrete. this should also be seen as one of the reasons for the high prevalence of fungi belonging to the genus cladosporium which readily colonise cement surfaces (e z e o n u et al. 1994; tr e a u , f e r n a n d e s c a l d a s 1994). apart from the genus cladosporium, fungi of the genera penicillium, aspergillus and alternaria are often recorded in residential buildings (g u t a r o w s k i et al. 2004). the latter was rarely recorded in the present study. this may be related to the fact that spores of this fungus are isolated from the atmospheric air, and its occurrence indoors is connected with inappropriately used mineral wool in building insulation (e z e o n u et al. 1994), the insulation method used in the schools studied. features that allow fungi to adhere quickly to the substrate or to occur in aerosols determine colonisation of indoor air or the retention of fungi in the air. this explains 252 e. ejdys the significant species difference between the walls and the air reported in the literature (g u t a r o w s k a et al. 2004) and observed in the present study. the discrepancy is related more to the formation of the surface of the spores than to their size. according to k r z y s z t o f i k (1992), particles whose diameter ranges between 3 and 10 microns may remain in the air for long periods of time, even at very low upward air currents. the occurrence of large and smooth ascospores of emericella quadrilineata on a smooth surface and only slightly smaller and rough conidia of its anamorph on a rough surface confirms these adaptations. a relatively permanent occurrence frequency of yeast-like fungi accompanied the great differences in the prevalence of moulds in the wall samples and in the air samples. it seems that while species predominant in indoor environment are mostly correlated with the properties of the building itself, the presence of yeasts and yeastlike fungi is correlated more with living organisms occupying it. this is shown by a great species correspondence of fungi isolated across the four seasons of the year from buildings and their occupants, recorded by m a c u r a and g n i a d e k (2003). indoor prevalence of yeasts and yeast-like fungi shown by these authors (10.7%) strictly corresponds to the present results. the occurrence of saccharomycopsis capsularis, a fungus recorded only since the late 1990s in the respiratory system in adults (d y n o w s k a , b i e d u n k i e w i c z 1999) and children (e j d y s 2003) in north-eastern poland, was recorded in the school buildings studied. the presence of yeast-like fungi is also affected by indoor humidity. their occurrence is reported in the second place after that of the genus penicillium in damp schools. additionally, indicator fungi of water-damaged buildings were isolated in such places: stachybotrys, trichoderma and a.versicolor (m e k l i n at al. 2002). this tendency was confirmed in the present study. only 10% prevalence of yeast-like fungi and 3% prevalence of a.versicolor were recorded at low air humidity in the schools. the richness of indoor mycobiota also results from the parallel sample incubation at different temperatures. it was accepted that while the temperature range of fungi is very broad it consists of narrow ranges of individual species. aspergillus fumigatus, a thermophilous fungus, with slow initial growth, may be an example. although it can grow at room temperature, it may lose competition with other, more rapid-growing species characteristic of this temperature. the results of the present author’s studies clearly show this. the domination of the genus aspergillus similar to that observed in the school buildings was recorded in a significantly warmer climate in student halls in egypt ( m a g h a z y , s h a a b a n , e l k a t a t n y 1 9 9 6 ) ; it was, however, isolated rarely (k r a w c z y k , k o w a l s k i , o c h ę c k a s z y m a ń s k a 1999) or was not isolated at all (g u t a r o w s k a at al. 2004) in flats in poland, which is another argument in favour of the expansion of the range of incubation temperatures to achieve greater knowledge of the mycobiota of various buildings. fungi in school buildings 253 conclusions 1. a total of 379 fungal isolates belonging to 32 genera of moulds, yeasts and yeast-like fungi were obtained in the school environment. the following genera dominated: aspergillus, penicillium and cladosporium. 2. of the 72 fungal species determined, cladosporium herbarum, aspergillus fumigatus and penicillium chrysogenum occurred most frequently in the school buildings. 3. wall surfaces were characterised by an increased prevalence of mycobiota in comparison with the air in these buildings, with only slightly greater species diversity. 4. a certain species specificity for rough and smooth wall surfaces was shown. fungi with large spores of the genera cladosporium and emericella adhered better to smooth surfaces while fungi with smaller conidia of the genus aspergillus – to rough surfaces. 5. the application of three incubation temperatures helped determine a fuller range of fungi colonising the school buildings. references b a r n e t t j. a., p a y n e r., ya r r o w d. 1990. yeasts: characteristics and identification. cambridge univ. press. b o g a c k a e . 1997. choroba budynków. mikol. lek. 4 (4): 233–237. b o u c h a r a j.p., d e c l e r c k p., c i m o n b., p l a n c h e n a u l t c., d e g e n t i l e l., c h a b a s s e d . 1996. routine use of chrom agar candida medium for presumptive identification of candida yeast species and detection of mixed fungal populations. clin microbiol. infect. 2 (3):202–208. b u d a k a . 1998. mikotoksyny. 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the genus aspergillus. wilians, wilkins, baltimore. r a p e r k.b., t h o m c., f e n n e l l d.i. 1949. a manual of the penicillia. wilians, wilkins, baltimore. s h e l t o n b.g., k i r k l a n d k.h., f l a n d e r s w.d., m o r r i s g.k. 2002. profiles of airborne fungi in buildings and outdoor environments in the united states. appl. environ. microbiol. 68 (4): 1743– 1753. to p b a s m., to s u n i., c a n g., k a k l i k k a y a n., a y d i n f. 2006. identification and seasonal distribution of airborne fungi in urban outdoor air in an eastern black sea turkish town. turk. j. med. sci. 36:31–36. tr u d e a u w., f e r n a n d e s c a l d a s e. 1994. identifying and measuring indoor air biologic agents. j. allergy clin. immunol. 94 (2): 393–400. z y s k a b . 2001. grzyby powietrza wewnętrznego w krajach europejskich. mikol. lek. 8 (3/4): 127–140. grzyby izolowane z pomieszczeń szkolnych s t r e s z c z e n i e celem badań było określenie składu gatunkowego grzybów występujących na powierzchniach ścian i w powietrzu budynków szkolnych. materiałem do badań były grzyby uzyskane z powietrza metodą sedymentacji i ze ścian techniką wymazu powierzchniowego, biorąc pod uwagę różne materiały wykończeniowe ścian. próby pobierano z wybranych pomieszczeń dwóch szkół: sali lekcyjnej, korytarza, toalet damskiej i męskiej, szatni ogólnej, szatni sportowej oraz prysznica. badanie przeprowadzono w maju 2005r., po zakończonym okresie grzewczym. grzyby hodowano na podłożu chapek-doxa równolegle w trzech temperaturach: 25, 37 i 40°c, przez co najmniej trzy tygodnie. w środowisku szkolnym z 321 prób uzyskano 379 izolatów grzybów z 32 rodzajów grzybów pleśniowych, drożdży i grzybów drożdżopodobnych. najczęściej izolowano rodzaje: aspergillus, penicillium i cladosporium. z oznaczonych 72 gatunków grzybów najczęściej w pomieszczeniach szkolnych występował: cladosporium herbarum, aspergillus fumigatus i penicillium chrysogenum. powierzchnie ścian charakteryzowały się zwiększoną prewalencją mikobioty w stosunku do powietrza tych pomieszczeń, przy nieco większej różnorodności gatunkowej. wykazano pewną specyficzność gatunkową grzybów dla szorstkich i gładkich powierzchni ścian. do powierzchni gładkich lepiej przylegały grzyby o dużych zarodnikach z rodzaju cladosporium i emericella, natomiast do szorstkich o mniejszych konidiach, z rodzaju aspergillus. zastosowanie trzech temperatur inkubacji pozwoliło na nakreślenie pełniejszego obrazu mikobioty środowiska szkolnego. 2014-01-01t11:46:35+0100 polish botanical society dedicated to professor barbara gumińska, an outstanding mycologist, academic teacher, cheerful and warm-hearted person, on the occasion of her eighty-fifth birthday p ho t. a rc hi w um professor barbara gumińska in the period of her academic activity professor barbara gumińska her life and achievements władysław wojewoda w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków childhood and education barbara apolonia lubelska was born on september 25th, 1924 in lwów (now lviv in ukraine). at the age of four she lost her father, who died in 1928. in lwów she attended primary school, and after 6 years she continued her education in the a. asnyk secondary school, until 1939. the next two years of education, namely the 8th and 9th grades, she continued in the secondary school no. 19 in lwów, in that period being under soviet authority. as under german occupation of lwów from 1941 all teaching was forbidden, young barbara lubelska was forced to work in the cosmetic laboratory “galicol”. attending secondary school was possible again after the germans’ withdrawal, when the city was taken by russians. she returned to the school no. 19 and continued her education in the 10th grade. however, after 9 months she left lwów with her mother and, as a repatriate, came to kraków in may 1945. in 1946, after completing her education in the j. joteyko secondary school, she graduated and became a student of the faculty of mathematics and natural sciences of the jagiellonian university in kraków. she graduated from the university in december 1951 and became a master of philosophy with the speciality in botany. work and scientific career as mentioned above, barbara lubelska started to work quite early in lwów under german occupation. in her 4th year of study at the jagiellonian university she was employed at the pedagogical university as an assistant, for the period of one year. in september 1950, still being a student, she was employed in the institute of botany of the jagiellonian university, where she was working until she retired in 1997. on june 3rd, 1954 she married jan gumiński. since then she published for a short time under the name lubelska-gumińska, and then under the name gumińska. in the acta mycologica vol. 45 (1): 3–10 2010 4 w. wojewoda years 1956-1959, besides working at the university, she was additionally a part-time employee in the institute of botany of the polish academy of sciences in kraków. at the jagiellonian university she was first working as assistant (1950-1953) and later senior assistant (1953-1963). during this time, in march 1962, she defended her phd thesis “mycoflora of the beech forests of rabsztyn and maciejowa”. in the years 1963-1972 she was working as postdoc (adiunkt), and in the years 1972-1977 as senior lecturer. in december 1974 she was awarded the doctor of science degree (habilitation), after presenting the work entitled “macromycetes of meadows in the pieniński national park”. in the years 1977-1985 she was employed as assistant professor and in the years 1985-1994, after being awarded a professorship, as professor. she retired in 1994 but was still a part-time employee until 1997. all together, she was a member of the university staff for 47 years. research and scientific achievements the main subject of the research carried out by barbara gumińska was taxonomy, ecology and geographical distribution of macrofungi (macromycetes), which belong to ascoand basidiomycota. she was working mainly in southern poland, e.g., on the kraków-częstochowa upland, and in polish carpathians: beskid niski mts, beskid sądecki mts, bieszczady mts, gorce mts and pieniny mts (especially pieniny national park). during her research on ecology and mycosociology she investigated mainly the fungi of beech, beech-fir and linden-hornbeam forests, and mountain meadows. especially interesting was a comparative study of the fungi occurring in the beech forests (dentario enneaphyllidi-fagetum) on the kraków-częstochowa upland, and beech-fir forests (dentario glandulosae-fagetum) in the beskid niski mts. she also participated in the research on mycobiota of the bieszczady zachodnie mts (polish east carpathians). in tyniec forests (kraków) she investigated the fungi of lindenhornbeam forest (tilio cordatae-carpinetum betuli). the most important, however, seems to be her work on the mycobiota of the pieniny mts and, especially, the pie-pieniny national park. in this part of her research she concentrated on the diversity of macrofungi, their ecology and phenology in the forest and meadow associations. thanks to her scientific activity, the pieniny national park is one of the best-investigated areas in poland when fungi are concerned. in her ecological works, she also paid attention to morphology and taxonomy of fungi and used to include her own drawings and pictures. one of the most important achievements of barbara gumińska is a well illustrated monograph of the fungi from the family hygrophoraceae occurring on the territory of poland and adjacent areas, published in the series “flora of poland – fungi”. during her research on fungi she also discovered several species new to poland or very rare in our country (szafer 1964). in 1956 she recorded a rare fungus – wawelia regia in the university botanical garden, for the first time after it was described as a new species in this location in 1908 by namysłowski. till now, this is the only known location of this possibly exotic species, which most probably arrived to the botanical garden with tropical plants. also the ecology of ring-forming macrofungi was the subject of her research, and this was the first such professor barbara gumińska. her life and achivements 5 detailed study on this phenomenon in poland. after a fire that destroyed vast forest areas in southern poland, she carried out the investigation of post-fire fungi in these places. together with dr. zofia heinrich she studied the collection of arctic and antarctic fungi brought by prof. m. olech from her expeditions. didactical achievements, popularization of mycology barbara gumińska was a very active university teacher (fig. 1). she held several courses and monographic lectures, seminars, practices and laboratory practicals with students in the areas of plant and fungal taxonomy and ecology. she also organized summer schools for the students of the jagiellonian university. many students and young scientists from other universities and scientific institutions in poland were attending trainings and consultations under her supervision and tutorial. she was the supervisor of two doctoral dissertations and several master degree theses in the area of mycology and lichenology. she was also a reviewer of numerous habilitation works, doctoral dissertations and master degree theses. in her popular science publications she promoted mycology. her papers concerning illustrations of fungi on post stamps are the first ones published in poland dealing with these artifacts. in 1965 she was one of the organizers of the great fungal exhibition held at the higher school of agronomy (now agricultural university) in kraków, entitled: “fungi in nature and economy” (“grzyby w przyrodzie i gospodarce człowieka”). fig. 1. course of plant systematics, the laboratory classes. from left: b. gumińska, a. stengl. 6 w. wojewoda functions and memberships of scientific societies barbara gumińska fulfilled important functions. she was the secretary of the scientific board of the institute of botany of the jagiellonian university (1976-1988), member of the biological commission of the polish academy of sciences (19761998), head of the department of plant taxonomy and phytogeography of the institute of botany (1977-1979), head of the mycology unit of the institute of botany (1979-1994), member of the editorial board of acta mycologica (1985-1998), member of the scientific board of w. szafer institute of botany of the polish academy of sciences (pas) in kraków (1990-1992), and member and vice-chairman of the commission for awards and distinctions of the faculty of biology and earth sciences of the jagiellonian university (1991-1993). periodically she was also a member of the commission for the habilitation courses of the faculty of biology and earth sciences, and vice-chairman of the council of the foundation for polish botany in the w. szafer institute of botany of pas. she is also a member of the mycological section of the polish botanical society. international cooperation, participation in international congresses and symposia barbara gumińska cooperated with several european mycologists, e.g., from former czechoslovakia. in 1956 she studied fungi under the supervision of a famous czech mycologist dr. a. pilát in the national museum in prague. in 1960 she participated in the 2nd congress of european mycologists in prague, and in two dreiländer mykologische tagung meetings: in hungary in 1964 and in austria in 1965. in 1966 she was one of the organizers of the 4th congress of european mycologists held in poland (fig. 2). published books and atlases together with władysław wojewoda, barbara gumińska prepared and published the first polish textbook on the identification of macrofungi entitled “grzyby owocnikowe i ich oznaczanie” (macrofungi and their identification). the first edition of this very popular book appeared in 1968, had three re-editions (under a modified title “grzyby i ich oznaczanie” – fungi and their identification), and was printed in 90 000 copies. on the origin of the book writes leńkowa (1992). for a long time it was the basic handbook, used both in schools and universities, to identify macrofungi. it was also widely used by hobbyists and amateurs of mushroom picking, and had a great impact on the popularization of the knowledge of fungi in poland. after several years of investigations on the fungi of the pieniny national park she published a colour atlas of macromycetes occurring in this interesting carpathian range (“atlas grzybów pienińskiego parku narodowego”). professor barbara gumińska. her life and achivements 7 students, the scientists a number of students of barbara gumińska followed her scientific career as mycologists working in universities and other scientific institutions. dr. anna drozdowicz. member of the department of plant taxonomy and phytogeography of the institute of botany of the jagiellonian university. phd student of b. gumińska. her research activity includes the taxonomy and ecology of slime moulds (myxomycetes). she is an author of the monographs of myxomycetes of popradzki landscape park and ojcowski national park, and co-author of the “checklist of polish myxomycetes” and the red list of polish myxomycetes. dr. anna ronikier. msc student of b. gumińska, now employed in the department of mycology of w. szafer institute of botany of the pas in kraków. in her research she deals with the diversity and ecology of slime moulds and macrofungi (mostly agaricoid and boletoid genera) of the high mountains in europe (mainly carpathians and pyrenees) and in south america. the author and co-author of several important articles and monographs, including the “checklist of polish myxomycetes”. prof. katarzyna turnau. msc and phd student of b. gumińska. for many years employed in the institute of botany (after retirement of b. gumińska the head of the mycological unit), now professor in the institute of environmental sciences of the jagiellonian university. world-class specialist in mycorrhiza research and mycology, author and co-author of many important scientific papers published in leading botanical and mycological journals, as well as book chapters dealing with fig. 2. excursion during the 4th congress of european mycologists, 1966. from left: m. ławrynowicz, j. kućmierz, b. gumińska, i. hołownia. 8 w. wojewoda taxonomy, ecology and ecophysiology of mycorrhizal and other fungi, especially the interactions of fungi and mycorrhizal plants with heavy metals. member of the polish academy of sciences. prof. władysław wojewoda. the first msc student of b. gumińska. although nominally the supervisor of the thesis was prof. b. pawłowski, b. gumińska in fact supervised the project. employed in the institute of botany of the jagiellonian university for 10 years, from 1969 until his retirement in 2003 employed in the w. szafer institute of botany of the pas, head of the department of mycology in this institute. founder of the biggest collection of fungi in poland, author and co-author of several papers dealing with taxonomy and ecology of macrofungi, e.g., the monograph of polish tremelloid and auricularioid basidiomycota, and the “checklist of polish larger basidiomycetes”, first editor of the “atlas of geographical distribution of fungi in poland”. scientific expeditions barbara gumińska was a very active member of the scientific field trips to the mycologically most precious areas in poland. the trips were organized and led by prof. alina skirgiełło from the warsaw university. in 1955 b. gumińska together with other polish mycologists (a. skirgiełło, m. lisiewska and a. nespiak) carried out the research on the biota of macrofungi in beskid sądecki, gorce and pieniny mountains (skirgiełło 1957, 2006). in may 1959 prof. skirgiełło led an expedition to puszcza białowieska primeval forest. barbara gumińska was again a member of the team, and i accompanied her in this expedition as a young student. thanks to her i was able to visit the most important wildlife area in poland for the first time, but she also introduced me to the elite of the polish mycologists: a. skirgiełło, s. domański, m. lisiewska, a. nespiak and w. truszkowska. i remember this visit very well; we used a horse carriage and were taking great delight in amazing fungal richness of that beautiful primeval forest. however, possibly the most important for the polish mycology were four expeditions to western bieszczady (polish eastern carpathians) organized in the years 1958-1965. barbara gumińska took part in the first three of them: to wetlina, ustrzyki górne and baligród (fig. 3). in that time the area of bieszczady was a blank spot on the mycological map of poland. before the world war ii this area was densely populated; however, after the war and displacement of indigenous inhabitants, it became almost empty and wild. many villages, e.g., ustrzyki górne, were only accessible by horse carriages or on foot, with food carried in the backpack. in this village, we were living in the only house that survived the war, the former military station. not infrequently during the whole day we did not meet a single man when walking around the mountains, looking for fungi and enjoying the beauty of the wild nature. thanks to these expeditions, the bieszczady mts were, for a long time, one of the best mycologically investigated areas in poland. the first expedition to wetlina in 1958 was described by skirgiełło (2006); the photos from the expeditions to the białowieża primeval forest and to bieszczady mts, in which b. gumińska took part, were published by wojewoda (1997). professor barbara gumińska. her life and achivements 9 scientific evaluations barbara gumińska prepared several scientific evaluations for many institutions and for industry, e.g., the department of forensic medicine in kraków, the clinic of children illnesses of the medical academy in krakow, and the factory of food concentrates in opole. awards and distinctions because of her scientific and didactic achievements barbara gumińska was given several awards and high distinctions: golden cross of merit for 20 years’ teaching activity at the jagiellonian university (1974), knight’s cross of polonia restituta for 30 years of outstanding teaching and educational activity (1983), third grade team award of the minister of higher education for excellent textbook for students (1969), third grade individual award for the achievements in scientific research for dsc dissertation (1978), second grade team award of the minister of science, higher education and technology for excellent textbook for students (1984), individual award of the minister of science and higher education for the monograph of hygrophoraceae (1998). also, professor gumińska was awarded and honoured by the vice-chancellor of the jagiellonian university several times. fig. 3. a. skirgiełło and b. gumińska during an expedition to western bieszczady, baligród 1960. 10 w. wojewoda outstanding mycologist, great educator, kind person summarizing the accomplishments of professor barbara gumińska it should be emphasized that she contributed greatly to the polish mycology as the author and co-author of many valuable scientific publications, popular science articles and textbooks. during her years at the university she educated many biologists and mycologists. a number of her students continue mycological research. she was very active and successful in propagating mycology among general public. barbara gumińska is also a very kind and friendly person, always calm, composed, and ready to offer help and advice. on the occasion of her 85th birthday we all wish professor barbara gumińska good health and further scientific achievements. acknowledgments. i wish to thank dr. piotr mleczko for his help in the work on this biography. references leńkowa a. 1992. profesor władysław szafer. anegdoty, fakty, wspomnienia. wydawnictwo i drukarnia „secesja”, kraków, 291 pp. (in polish). skirgiełło a. 1959. notatki mikologiczne z okolic krościenka nad dunajcem. monogr. bot. 8 (2): 229–235 (in polish). skirgiełło a. 2006. zapiski ze stuletniego życia. bel studio sp., warszawa, 151 pp. (in polish). szafer w. 1964. zarys historii botaniki w krakowie. uniwersytet jagielloński. wydawnictwa jubileuszowe 19:1–169 (in polish). wojewoda w. 1997. life and works of professor stanisław domański. wiadomości botaniczne 41 (1): 39–47 (in polish). all photos: archive of prof. b. gumińska. 2014-01-01t11:50:00+0100 polish botanical society biodiversity of zoosporic fungi in polluted water drainages across niles´ delta region, lower egypt esam h. ali botany department, faculty of science, assiut university assiut, egypt, ibraheem55@yahoo.com a l i e. h.: biodiversity of zoosporic fungi in polluted water drainages across niles’delta region, lower egypt. acta mycol. 42 (1): 99-111, 2007. thirty-four identified in addition to five unidentified species appertaining to ten genera of zoosporic fungi were identified and isolated from eighty four polluted water samples, which were randomly collected from different polluted sites of the water drainages along the niles delta in lower egypt. baiting sesame seeds culture technique was employed at 20±2°c for the recovery of zoosporic fungi. the genera; pythium and saprolegnia (8 and 7 zoosporic fungal species, respectively) showed the broadest spectra of species diversity whereas aqualinderella was only represented by one species (a. fermentans). saprolegnia delica and dictyuchus carpophorus (the greatest fungal populations) were the most dominant isolated zoosporic fungal species where they were highly occurred especially at the hyperpolluted waters with the heavy metals. these two species could be considered as indicators for the response of the structure and function of microbial communities for water pollution. several zoosporic fungal species were rarely encountered. both aqualinderella fermentans and pythium rostratum were recovered in moderate frequency of occurrence. water samples which had high concentrations in heavy metals were the poorest in the species diversity of zoosporic fungi. despite that, fungal species belonging to the family saprolegniaceae flourished in hyper polluted water samples whilst those belonging to the family pythiaceae predominated in more diluted water samples. also, the prevalent species; s. delica and d. carpophorus were not affected by heavy metals concentrations being as indicators for water pollution with the heavy metals. ph values of the polluted water samples had no influence on the occurrence of zoosporic fungi. water samples characterized by high organic matter content and low total soluble salts were the richest in zoosporic fungal species. key words: zoosporic fungi, diversity, pollution, drainages introduction heavy metals pollution has increased over the last few decades through mining, industrial emissions and garbage disposal, and as by-products of agricultural fertilizers (m e r i a n 1991). as a result pristine ecosystems are becoming scarce. the acta mycologica vol. 42 (1): 99-111 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday biodiversity of zoosporic fungi 101 ples were brought directly to the laboratory for the recovery of zoosporic fungi and chemical analysis. physico-chemical analysis of polluted water samples. physical characteristics of water samples such as water temperature and ph value were measured in situ with field probes (thermometer and ph meter). glass bottles containing water samples with no added sesame seeds (bottles no. 6) were used for the chemical analysis at each sampling site. the total soluble salts and organic matter contents were chemically estimated based on j a c k s o n (1958). the heavy metals (pb, zn, co, cu and cd) of polluted water samples were determined with an atomic absorption spectrophotometer (varian, model aa 55). microbial analysis for baiting and recovery of zoosporic fungi. for baiting and recovery of zoosporic fungi, the polluted water samples containing the baits of sesame seeds were poured under aseptic conditions into petri-dishes (15 cm in diameter). petri-dishes (5 for each water sample) were transferred into an incubator at 20°c and left over night (24 hours) during the colonization of sesame seeds by fungal propagules (e l h i s s y , k h a l l i l 1989). colonized sesame seeds were transferred into other equivalent, smaller petri-dishes (10 cm in diameter each) to which crystalline penicillin (2000 units/ ml) was supplied for suppressing the bacterial growth (r o b e r t s 1963). the dishes were then, again, incubated at 20°c for a period of twenty days during which they were examined daily for the identification of zoosporic taxa. transferring colonized sesame seeds into new petri dishes with sterilized distilled water refreshed growth of taxa of zoosporic fungi. most of the oomycetes were purified on glucose-peptone (gp) agar medium (w i l l o u g h b y , p i c k e r i n g 1977) whereas yeast-peptone starch medium (ypss) was used for purification of allomyces sp. (e m e r s o n 1941), which belong to chytridiomycetes. the fungal cultures were maintained on the previously mentioned media and stored at 8-12°c and sub-cultured every 2-3 months. the number of cases of isolation and occurrence remarks of each taxa (genera and species) of zoosporic fungi was calculated from the total number of the tested polluted water samples (84 water samples). they were designed as follows: h= high occurrence; more than 42 samples out of 84 water samples m= moderate occurrence; between 21-42 water samples l= low occurrence; between 10-20 water samples r= rare occurrence; less than 10 samples out of 84 water samples. regarding the determination of total counts of zoosporic fungal genera and species which were recovered during this study, the fungal species appeared on one dish was counted as one colony to the number of seeds (40 isolates) for each tested sample. identification of the genera and species of zoosporic fungi. the following references were adopted as syllabi for the morphological identification of the genera and species of zoosporic fungi during this study and they were as follows: c o k e r (1923), j o h n s o n (1956, 1971, 1977), wa t e r h o u s e (1956, 1968), s p a r r o w (1960), s c o t t (1961), s e y m o u r (1970), k a r l i n g (1977) and n a t v i g (1987). statistical analysis. simple and multi-correlation statistical analysis was designed. this statistical analysis was functioned to test the effect of physicochemical characteristics of water samples versus the number of the recovered species of zoosporic fungi in each water sample. 102 e. h. ali results and discussion the ph values of the polluted water samples, which were collected from different water drainages located in nile delta region in lower egypt, were in the alkaline side and ranged between 8.36 and 11.15 (tab. 1). the results of the simple and multiple correlation analysis showed that the ph value had no role in the biodiversity of the isolated zoosporic fungi. other authors have found that ph did not governor the distribution and occurrence of zoosporic fungi in different water habitats. g o p t a and m e h r o t r a (1989) observed that ph value was insignificant factor affecting the occurrence of zoosporic fungi. despite that, l u n d (1934) found that a number of aquatic fungi were specific for highly acidic waters, some being fairly common and some quite rare. in addition, k h u l b e (1980a), who studied the influence of ph on the distribution of zoosporic fungi in some lakes of nainital region, india concluded that the incidence of zoosporic fungi was related to ph. he found high fungal population in lakes with ph of 7.3 to 8.8. s m i t h et al. (1984) mentioned that ph has complex effects not only on the activity of zoospores, mycelial growth, enzyme activity and reproduction, but also affects the availability of salts such as calcium, potassium, iron and phosphorus and the form in which nitrogen is present. also, s h e a r e r and we b s t e r (1985) reported that the more acidic (ph 5.4-6) upstream portion of the river teign had an impoverished mycobiota when compared with less acidic (ph 7-7.2) downstream portion. other authors (d u b e y 1990) reported more taxa of zoosporic fungi in alkaline waters although freshwater forms can occur across a wide ph range. moreover, d u b e y et al. (1994) isolated more taxa of zoosporic fungi from six steams, located on or near the fernow experimental forest in tucker county in west virginia, with high ph than those from low ph. the estimated total soluble salts of the polluted water samples fluctuated between 474 and 3200 mg/ l (tab. 1). it had an inverse correlation with the number of the recoverable zoosporic fungal species from the collected water samples (p ≤ 0.001). similar results were also obtained by h a r r i s o n and j o n e s (1974) who found that higher salinities limited the distribution of these fungi. in contrast, r a t t a n et al. (1980) indicated that salinity played a non-significant role in the occurrence of saprolegniaceae recovered from water samples in shatt al-arab, iraq. g l e a s o n et al. (2006) reported that several species of chytridiomycota could survive various salt content. some other investigators (te s t r a k e 1959; e l h i s s y , k h a l l i l 1989) found that some species of zoosporic fungi could withstand a certain degree of salinity, inasmuch as they live in brackish water of estuaries. organic matter content of the polluted water samples varied from 150.47 to 497.10 mg/ l (tab. 1). the result showed that the organic matter content of water drainages was positively correlated with the number of the recoverable species of zoosporic fungi (p ≤ 0.001). accordingly, c a n t i n o and tu r i a n (1959) found that some species of saprolegniales usually depend on the presence of organic compounds for nutrition. in addition, misra (1982) reported that the presence of organic debris is one of the probable reasons for the frequent and higher collections of zoosporic fungi from ponds having more vegetation and hence more organic matter content. similar results were also obtained by r o b e r t s (1963) and o k a n e (1978). k h u l b e (1981) and e l h i s s y et al. (1992) reported a significant correlation between the incidence biodiversity of zoosporic fungi 103 of zoosporic fungi and organic matter content. however, r a v i r a j a et al. (1998) reported a decline in aquatic hyphomycete diversity with increased organic pollution. analysis of the polluted water samples for the content of heavy metals (tab. 1) showed that the value of the heavy metal cd fluctuated between 0.0082 and 0.0177 ppm, for pb it was ranged between 0.0803 and 3.3845 ppm, for co 0.0010 to 0.0464, for cu 0.0292 to 0.0540 and for zn 0.0377 and 0.1792 ppm. these values for the heavy metals are higher with multiples than that of water samples obtained at the same unpolluted sites taken from the river nile as reported by s o l t a n and a w a d a l l a h (1995). there was a parallel increment between the values of these estimated heavy metals within one sample. as the concentrations of the heavy metals in water drainages increased the number of zoosporic fungal species decreased and vice versa (p ≤ 0.001). these results were also similar for several findings in different taxonomic fungal groups. in this connection, m a l t b y and b o o t h (1991) and b e r m i n g h a m et al. (1996) reported that coal amine effluent which rich in heavy metals reduced the number of aquatic hyphomycete species by 15 to 39% compared with relatively clean, undisturbed streams. also, g a n o v a s et al. (2003) isolated a filamentous fungus (aspergillus sp. p37) from the tinto river in spain, with its high acidity and heavy metal concentration, able to grow at 15000 ppm of arsenic. they claimed that the fungus was capable of removing arsenic from culture media. in addition, k r a u s s et al. (2003) isolated several aquatic hyphomycetous species from polluted groundwater wells located in a former copper shale-mining district (11 sites; mansfelder land, central germany) and in the mulde and elbe rivers (2 sites). these locations have relatively high levels of pb, mn, and fe. they found that heliscus lugdunensis and anguillospora sp. were the most widespread species. the results presented in table 2 indicate that thirty four identified in addition to five unidentified species which belong to ten genera of zoosporic fungi were recovered and isolated from eighty four polluted water samples, which were collected at random from different sites at the polluted drainages along niles´ delta in lower egypt. pythium and saprolegnia were represented by 8 and 7 species, respectively ta b l e 1 fluctuations of ph values, total soluble salts (tss; mg/ l), organic matter content (omc; mg/ l) and the heavy metals cd, pb, co, cu and zn (ppm) of the collected polluted water samples (pws) from the different drainages at seven districts in the nile delta area in lower egypt district pws ph tss omc cd pb co cu zn elkalyobia 1-12 8.4810.58 5371664 150.47189.43 0.01310.0149 0.08031.2778 0.02330.0252 0.03410.0369 0.04340.0475 elmonofeyia 13-24 9.3511.15 474768 163.91228.40 0.00820.0089 0.14260.3033 0.00100.0141 0.02960.0305 0.04170.0534 elgharbia 25-36 10.3810.60 8961984 176280.79 0.01550.0177 3.12123.3845 0.01460.0417 0.03750.0441 0.05410.1755 kafr elshekh 37-48 10.1511.12 10242423 189.43202.87 0.01640.0167 2.72483.2998 0.01610.0352 0.04100.0454 0.05860.1792 damietta 49-60 9.5810.38 21763200 163.91189.43 0.01440.0159 1.49562.1456 0.01260.0164 0.03010.0421 0.03770.0381 eldakahlia 61-72 10.4711.15 6401088 214.96241.83 0.01520.0154 2.25183.1750 0.02230.0464 0.03590.0540 0.04820.0528 el-sharkya 73-84 8.3610.71 524832 228.40497.10 0.00820.0142 1.06471.0755 0.00140.0092 0.02920.0379 0.04200.0545 104 e. h. ali and thus they comprised the broadest spectra of the species amongst the different genera of zoosporic fungi (fig. 2). saprolegnia delica (fig. 4) and dictyuchus carpophorus (fig. 5) were the most prevalent zoosporic fungal species (tab. 2) during this study where they were of high frequency of occurrence (fig. 3). these species predominated in the hyper-polluted waters with the heavy metals at the different investigated drainages in the delta of nile. this result was not expected and these ta b l e 2 number of cases of isolation (nci), total counts and occurrence remarks (or) of zoosporic fungi recovered from 84 polluted drainages water samples in the niles´ delta region, lower egypt using water sesame seeds baiting technique zoosporic fungal genera and species n. c. i. total counts % of total counts o. r. achlya total a. conspicua coker a. dubia coker a. glomerata coker a. orion coker & couch a. proliferoides coker achlya sp. allomyces total a. anomalus emerson a. macrogynus emerson & wilson aqualinderella fermentans emerson & weston aphanomyces total a. helicoids von minden a. laevis de bary a. stellatus de bary aphanomyces sp. brevilegnia total b. unisperma coker & braxton brevilegnia sp. dictyuchus total d. carpophorus zopf. d. magnusii lindst. d. sterilis coker phytophthora total p. cactorum (lebert & cohn) schroeter p. cinchonae sawada p. cinnamomi rands p. cryptogea pethybridge & lafferty p. inflata caroselli & tucker pythiopsis cymosa de bary pythium total p. debaryanum hesse p. irregulare buisman p. proliferum de bary p. rostratum butler p. spinosum sawada p. ultimum trow p. vexans de bary pythium sp. saprolegnia total s. delica coker s. diclina humphrey s. ferax (gruith.) thuret s. furcata maurizio s. hypogyna (pringsheim) de bary s. uliginosa johannes saprolegnia sp. 54 6 12 4 3 23 6 24 8 16 26 25 5 12 6 2 8 6 2 58 47 2 9 37 3 13 11 8 2 15 41 3 4 2 21 2 3 4 2 68 49 3 11 5 2 4 1 445 49 82 32 24 186 72 261 76 185 115 206 18 152 31 5 59 38 21 383 268 26 89 155 12 58 52 28 5 62 276 14 26 6 194 5 14 11 6 426 254 16 87 23 12 19 5 18.63 2.05 3.43 1.34 1.01 7.79 3.02 10.93 3.18 7.75 4.82 8.63 0.75 6.37 1.30 0.21 2.47 1.59 0.88 16.04 11.22 1.09 3.73 6.49 0.50 2.43 2.18 1.17 0.21 2.60 11.56 0.59 1.09 0.25 8.12 0.21 0.59 0.46 0.25 17.84 10.64 0.67 3.64 0.96 0.50 0.80 0.21 h r l r r m r m r l m m r l r r r r r h h r r m r l l l r l m r r r m r r r r h h r l r r r r total number of isolates 2388 106 e. h. ali in an interpretation for the prevalence of some species of aquatic hyphomycetes and other fungal groups in heavy metals polluted habitats; g a d d (1993), m i e r s c h et al. (1997, 2001) and g a d d and s a y e r (2000) suggested that these species have evolved some tolerance of heavy metals by synthesizing s-rich compounds and peptides derived from glutathione (phytochelatines). several species of the isolated zoosporic fungi were illustrated in figures 4-16. some representative structures of the isolated species of zoosporic fungi. saprolegnia was the leading zoosporic fungal genus during this study and it was of high occurrence where it was recovered from 68 polluted water samples encountering 17.84% of total count of isolates. it was represented by six identified and unidentified isolates of which s. delica highly occurred (49 water samples) counting 10.64% of total number of isolates. saprolegnia ferax was of low occurrence (tab. 2) whilst s. furcata, s. uliginosa, s. diclina, s. hypogyna and the unidentified species were rarely occurred as data shown in table 1. saprolegnia delica was isolated for the first time during this study and it was surprising to isolate it in high frequency of occurrence. hyper-polluted water samples of the investigated drainages were the richest in the species occurrence. in this regard, intensive researches on the occurrence of zoosporic fungi from natural water habitats in the river nile (e l h i s s y et al. 1982, 1992), in brackish water habitats in several egyptian lakes (e l h i s s y et al. 2004) and in polluted waters with industrial effluents of fertilizers (e l h i s s y et al. 2001) revealed that saprolegnia was also the major prevalent genus of zoosporic fungi. in addition, saprolegnia ferax was encountered in a number of places in poland including springs (c z e c z u g a et al. 1989), melting snow pools (c z e c z u g a 1992), sunk well water (c z e c z u g a et al. 1987) and forest stream (c z e c z u g a et al. 1986). moreover, m e h d i et al. (2001) investigated zoosporic fungi in polluted water which collected from pasni and ormara coasts of pakistan using baiting and plating techniques. they isolated saprolegnia diclina, s. parasitica and unidentified taxa of zoosporic fungi from these sites. dictyuchus came in the second position after saprolegnia during this study and it was also of high occurrence (tab. 2). it was included three identified species of which d. carpophorus was the most prevalent and highly occurred (tab. 2). the recoverable isolates and incidence of d. carpophorus increased at the hyper-polluted waters with the heavy metals. the remaining species were isolated in rare frequency of occurrence and they were d. sterilis and d. magnusii (tab. 2). dictyuchus was nearly recovered either in moderate or low frequency of occurrence in different egyptian water habitats including the river nile (e l h i s s y , k h a l l i l 1989; e l h i s s y et al. 1992, 2001, 2004; e l n a g d y , a b d e l h a f e z 1990). achlya was isolated in high frequency of occurrence (tab. 2) and it was matching 18.63% (445 isolates) of total number of isolates. it contributed five identified in addition to unidentified species of which a. proliferoides was isolated from 23 polluted water samples (moderately occurred) representing 7.79% of total number of isolates. the remaining genera were a. dubia, which was of low incidence (tab. 2), a. conspicua, unidentified species, a. glomerata and a. orion were rarely isolated (tab. 2). achlya was previously recovered from different water habitats in nile system and lakes in usa (z i e g l e r 1958), in nigeria (a l a b i 1973), in india (h a s i j a , b a t r a 1978; k h u l b e 1980b; m i s r a 1982), in iraq (r a t t a n et al. 1980) and in biodiversity of zoosporic fungi 107 egypt (e l h i s s y et al. 1982, 2004). it was contributed the broadest spectrum of species diversity in different aquatic habitats as reported by l u i and vo l z (1976), k l i c h and t i f f a n y (1985) and e l n a g d y and n a s s e r (2000). pythium was of moderate frequency of occurrence (tab. 2) counting 11.56% of total fungal isolates. it contributed the broadest spectrum of the isolated fungal species during this study where it was represented by seven identified in addition to unidentified species. of the isolated pythium species, p. rostratum was the most frequent where it was moderately occurred (tab. 2). the data presented in table 2 indicate that the rest of pythium species were of rare frequency of occurrence and they were p. irregulare, p. vexans, p. debaryanum, p. ultimum, p. proliferum, p. spinosum and pythium species. pythium came in the second position in the waters of the major lakes in egypt as indicated by e l h i s s y et al. (2004). it was also dominant in some ponds of kharga oases (e l n a g d y , a b d e l h a f e z 1990) and in waters polluted with industrial effluents of kima factory for fertilizers at aswan region, upper egypt (e l h i s s y et al. 2001). pythium was also recovered from water of the river biała in poland as reported by c z e c z u g a and m a z a l s k a (1996). however, it was recovered in rare incidence from rainfall water accumulated at the common valleys in the south-eastern region in saudi arabia (a l i , n a s s e r 2001). phytophthora was also recovered in moderate occurrence (tab. 2). the genus was represented by five species. three of these namely: p. cinchonae, p. cinnamomi, p. cryptogea were of low occurrence (tab. 2). the other two species p. cactorum and p. inflata were isolated in rare occurrence (tab. 2). phytophthora was also recovered in moderate incidence from surface water samples of major egyptian lakes (e l h i s s y et al. 2004) where it was contributed the broadest spectrum of species diversity. it was commonest in the lakes of hypertonic saline waters. aqualinderella was recovered in moderate frequency of occurrence and it was represented by only one species (a. fermentans, 26 polluted water samples, tab. 2). in this regard, aqualinderella fermentans was also repeatedly recovered in different incidences from different water habitats in egypt (e l h i s s y , k h a l l i l 1989; e l h i s s y et al. 2001, 2004) and saudi arabia (e l n a g d y , n a s s e r 2000). however, it was more common in stagnant than running waters in germany (e l h i s s y , o b e r w i n k l e r 1999). aphanomyces was also of moderate incidence (25 polluted water samples, tab. 2) and it was included three identified in addition to unidentified species. these species were: a. laevis which was of low occurrence (12 water samples enumerate in 6.37% of total fungal isolates), a. stellatus, a. helicoides and aphanomyces species which rarely occurred (tab. 2). the incidence of these species of aphanomyces was reported but in different frequencies from regional surface water of the river nile at rossetta and damietta branches, lower egypt (e l -h i s s y , k h a l l i l , 1989), at upper egypt districts (e l h i s s y et al. 1982, 1992), from water samples polluted with industrial wastes of kima factory for fertilizers at aswan region, south egypt (e l h i s s y et al. 2001) and from the surface water of the major egyptian lakes (e l h i s s y et al. 2004). in addition, e l n a g d y , n a s s e r (2000) recovered aphanomyces laevis in rare frequency from rainwater samples in the riyadh, saudi arabia. allomyces was moderately isolated from 24 polluted water samples matching 10.93% of total fungal isolates. this genus was represented by two species of which a. macrogynus was of low incidence (tab. 2) and a. anomalus, which rarely occurred 108 e. h. ali (tab. 2). similarly, c z e c z u g a and g o d l e w s k a (1994) isolated allomyces arbuscula from shallow lake in forests with acid water and low mineral salt content. however, e l h i s s y et al. (2004) isolated allomyces species (a. anomalus, a. macrogynus and a. moniliformis) in rare occurrence from the surface waters in major four egyptian lakes. pythiopsis: p. cymosa was of low frequency of occurrence where it was isolated from 15 polluted water samples constituting 2.60% of total fungal isolates. brevilegnia was isolated in rare frequency of occurrence where it was emerged from 8 polluted water samples comprising 2.47% of total fungal isolates. it included two species, b. unisperma and brevilegnia sp. which were of, low occurrence (tab. 2). in this regard, s t e c i o w (2003) isolated brevilegnia ensenadensis from a man made polluted channel near a petroleum refinery in buenos aires province in argentina. general outlook on the recoverable species of zoosporic fungi during this study indicated that the prevalence of saprolegnia delica and dictyuchus carpophorus. s. delica was isolated for the first time from egyptian water habitats and it was surprised to isolate it in 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(in:) k. d. h y d e , w. h o , s b. p o i n t i n g (eds). aquatic mycology across the millennium. fungal diversity 5: 119–129. s t e c i o w m.m. 2003. a new species of brevilegnia (saprolegniales, straminipila) from buenos aires provinces, argentina. mycologia 95: 934–942. biodiversity of zoosporic fungi 111 te s t r a k e d. 1959. estuarine distribution and saline tolerance of some saprolegniaceae. phyton 12: 147–152. wa t e r h o u s e g.m. 1956. the genus phytophthora. diagnosis (or descriptions) and figures the original papers. (in:) miscellaneous publication 12. comm. mycol. inst. kew, surrey, england, 120 pp. wa t e r h o u s e g.m. 1968. the genus pythium pringsheim. p. 1–71. (in:) mycol. pap. 110, comm. mycol. inst. kew, surrey, england. w i l l o u g h b y l.g., p i c k e r i n g a.d. 1977. viable saprolegniaceae spores on the epidermis of the salmonid fish salmo trutta and salvelinus alpinus. trans. br. mycol. soc. 68: 91–95. z i e g l e r a.w. 1958. new water molds from florida. mycologia 50: 693–696. figs 4-9. fig. 4. saprolegnia delica – oogonia with diclinous and androgynous antheridia; fig. 5. dictyuchus carpophorus – dictyoid-zoosporangia and eccentric oogonium with characteristic single oosphere; fig. 6. dictyuchus sterilis – showing only sympodial, dictyoid-shaped zoosporangia and lacking sex organs; fig. 7. dictyuchus magnusii – dictyoid-zoosporangium and sex organs; fig. 8. saprolegnia furcata – oogonium with coiled oogonial stalk; fig. 9. saprolegnia diclina – diclinous antheridium completely encircling an oogonium. 6 7 4 5 8 9 figs 10-16. fig. 10. pythiopsis cymosa – coenocytic hypha bearing single globular sporangium; fig. 11. saprolegnia ferax – spherical oogonia having no antheridial attaches; fig. 12. phytophthora cinchonae – proliferated, lemon-shaped zoosporangia; fig. 13. achlya proliferoides – antheridial branches closely twined around the vegetative hyphae and oogonia; fig. 14. allomyces macrogynus – gametophyte bearing gametangia in pairs with the epigynous male gametangia and the hypogynous female gametangia; fig. 15. pythium irregulare – plerotic oospore with distinguished irregulare surface; fig. 16. brevilegnia sp. – zoosporangium showing typical brevielegnoid discharge. 10 12 11 13 14 15 16 2014-01-01t11:45:31+0100 polish botanical society septoglomus deserticola emended and new combinations in the emended definition of the family diversisporaceae janusz błaszkowski and gerard chwat department of plant protection, west pomeranian university of technology słowackiego 17, pl-71-434 szczecin, janusz.blaszkowski@zut.edu.pl błaszkowski j., chwat g.: septoglomus deserticola emended and new combinations in the emended definition of the family diversisporaceae. acta mycol. 48 (1): 89–103, 2013. an updated morphology of spores of septoglomus deserticola, an arbuscular mycorrhizal fungus of the phylum glomeromycota, is presented based on the original description of the species, only one other its definition recently published and spores produced in pot cultures inoculated with the rhizosphere soil and root fragments of an unrecognized grass colonizing maritime sand dunes of the hicacos peninsula, cuba. phylogenetic analyses of sequences of the large subunit (lsu) nrdna region of the cuban fungus confirmed its affinity with s. deserticola deposited in the international bank for the glomeromycota (beg) and indicated that its closest relatives are s. fuscum and s. xanthium. phylogenetic analyses of sequences of the small subunit (ssu) nrdna confirmed the cuban fungus x s. fuscum x s. xanthium relationship revealed in analyses of the lsu sequences and thereby suggested the cuban septoglomus is s. deserticola. however, it was impossible to prove directly the identity of the cuban fungus and s. deserticola from beg based on ssu sequences due to the lack of s. deserticola ssu sequences in public databases. in addition, phylogenetic analyses of lsu and ssu sequences confirmed the uniqueness of the recently erected genus corymbiglomus with the type species c. corymbiforme (formerly glomus corymbiforme) in the family diversisporaceae and proved that its lsu sequences group in a clade with lsu sequences of g. globiferum and g. tortuosum. consequently, the two latter species were transferred to corymbiglomus and named c. globiferum comb. nov. and c. tortuosum comb. nov., and the definitions of the family diversisporaceae and the genus diversispora were emended. key words: arbuscular mycorrhizal fungi, glomeromycota, taxonomy acta mycologica vol. 48 (1): 89–103 2013 doi: 10.5586/am.2013.011 90 j. błaszkowski and g. chwat introduction arbuscular mycorrhizal fungi (amf) of the phylum glomeromycota are associated with ca. 70–90% of land plants (smith, read 2008; brundrett 2009), including those growing in extremely poor maritime sand dunes (koske 1987; dalpé 1989; tadych, błaszkowski 2000). it is recognized that maritime sand dunes especially favor the development of amf because of the low content of nutrients and organic matter (błaszkowski et al. 2009; błaszkowski 2012). the fungi frequently increase the supply of plants with nutrients and decrease their sensitivity to different abioand biotic stresses (schönbeck 1978; dehn, schüepp 1989; griffioen, ernst 1989; smith, read 2008). although even low colonization of plant roots by amg can alleviate such stresses (pongrac et al. 2007), the effect of influence of amf may differ, because different species or even strains of a given species of amf may variously affect plants (abbott, robson 1981; kaldorf et al. 1999; maherali, klironomos 2007; sýkorová et al. 2011). hence numerous unsuccessful attempts of application of amf probably partly resulted from erroneous species identification and the difficult nature of am fungal taxonomy (schüßler et al. 2011; krüger et al. 2012). at present the phylum glomeromycota comprises three classes, five orders, 15 families, 34 genera and ca. 250 species (oehl et al. 2011a-e; schüßler, walker 2010; błaszkowski 2012). however, results of molecular phylogenetic analyses of sequences of nrdna extracted from plant roots suggest that less than 5% of the existing species in the world are known to date and that most undescribed species form glomoid spores (krüger et al. 2009), i.e. spores similar in mode of formation, spore wall structure and in characters of their subtending hypha to those of glomus macrocarpum tul. & c. tul., the type species of the genus glomus tul. & c. tul. (schüßler, walker 2010). of the described species, ca. 62% produce typical glomoid spores. glomoid spores form pacispora spp. as well, but they have two spore walls (vs. one in typical glomoid spores), an outer wall, forming the spore surface, and an inner wall, called a germinal wall (błaszkowski 2012). apart from glomus spp., glomoid spores form fungi of 15 other genera, among which the recently erected genus septoglomus sieverd., g.a. silva & oehl is, whose type species is s. constrictum (trappe) sieverd., g.a. silva & oehl. (oehl et al. 2011c). currently the genus septoglomus comprises seven species whose spores are dark-coloured and have a 2– to 3–layered spore wall with the innermost layer being laminate (oehl et al. 2011c; błaszkowski et al. 2012). none of the layers stains in melzer’s reagent. the spore subtending hyphae of the species usually is cylindrical or constricted, and its pore is closed by a septum. we established single-species cultures from dark-coloured glomoid spores of the glomeromycota isolated from a trap culture inoculated with the rhizosphere soil and root fragments of an unrecognized grass colonizing maritime sand dunes of the hicacos peninsula, cuba. morphological studies of these spores suggested we found an amf closely related to s. deserticola (trappe, bloss & j.a. menge) g.a. silva, oehl & sieverd. apart from the original description of s. deserticola as g. deserticola (trappe et al. 1984), in the literature there is only one other report of morphology of the species prepared from spores obtained from the international bank for the glomeromycota (beg; błaszkowski 2012). our doubt of the identity of the two septoglomus deserticola emended and new combinations 91 fungi resulted from differences in colour of their spores; the spores from cuba were lighter. however, subsequent phylogenetic analyses of sequences of the large subunit (lsu, partial) spore nrdna region unambiguously placed the cuban fungus among available lsu sequences of s. deserticola. we also obtained sequences of the small subunit (ssu) nrdna of the fungus from cuba, but we could not confirm its identity to s. deserticola due to the lack of ssu sequences of s. deserticola in available databases. however, our phylogenic analyses indicated that the position of the cuban fungus in a tree with ssu sequences relative to other septoglomus spp. was identical to that in the lsu tree, suggesting the cuban septoglomus is s. deserticola. thus our ssu sequences significantly widened the range of molecular data on s. deserticola. phylogenetic analyses of lsu sequences of g. corymbiforme błaszk. recently lead to the erection of a new genus, corymbiglomus błaszk. et chwat, with the type species, c. corymbiglomus (błaszk.) błaszk. et chwat (błaszkowski 2012). subsequent analyses of lsu and ssu sequences of this species confirmed the uniqueness of corymbiglomus and its phylogenetic relationship to diversispora c. walker et a. schüssler, emend. g.a. silva, oehl et schüssler and indicated that other species grouping in a clade with lsu sequences of c. corymbiforme are g. globiferum koske et c. walker and g. tortuosum n.c. schenck et g.s. sm. thus results of the analyses and morphological similarity of spores of the tree species proved that g. globiferum and g. tortuosum should became members of the genus corymbiglomus. the aims of this paper are to update the knowledge on morphology and phylogeny of s. deserticola, emend the definitions of the family diversisporaceae and the genus diversispora and to transfer g. globiferum and g. tortuosum to the genus corymbiglomus. materials and methods establishment and growth of trap and single-species cultures, extraction of spores, and staining of mycorrhizae. spores examined in this study were derived from both pot trap and single-species cultures. trap cultures were established to obtain living spores and to initiate sporulation of species that may not have sporulated in the field collections (stutz, morton 1996). the method used to establish trap cultures, their growing conditions and the methods of spore extraction and staining of mycorrhizae were as those described previously (błaszkowski et al. 2012b). the growing substrate of trap cultures was the field-collected rhizosphere soil and roots of the plant species sampled mixed with autoclaved coarse grained sand. single-species cultures were also established and grown as given in błaszkowski et al. (2012b). briefly, the cultures of s. deserticola were successfully established from small clusters of spores (5–15) attached by a common mycelium. the growing substrate of the cultures was autoclaved commercially available coarse-grained sand (grains 1.0–10.0 mm diam 80.50%; grains 0.1–1.0 mm diam 17.28%; grains < 0.1 mm diam – 2.22%) mixed (5:1, v/v) with clinopthilolite (zeocem, bystré, slovakia) of grains 2.5–5 mm. clinopthilolite is a crystaline hydrated alumosilicate of alkali metals and alkaline earth metals having, e.g. high ion exchange 92 j. błaszkowski and g. chwat capability and selectivity, as well as reversible hydration and dehydration. the sand-clinopthilolite mixture had a ph of 7.3. the cultures were kept in transparent plastic bags, 15 cm wide and 22 cm high as suggested by walker and vestberg (1994). the cultures were watered with tap water once or twice a week, harvested after five months when spores were extracted for study. to reveal mycorrhizal root structures, root fragments located ca. 1–5 cm below the upper level of the growing medium were cut off with a scalpel. plantago lanceolata l. was used as host plant in both trap and single-species cultures. microscopy. morphological properties of spores and their wall structure were determined after examination of at least 100 spores mounted in water, lactic acid, polyvinyl alcohol/lactic acid/glycerol (pvlg; omar et al. 1979) and a mixture of pvlg and melzer’s reagent (1:1, v/v). spores at all developmental stages were crushed to varying degrees by applying pressure to the cover slip and then stored at 65o c for 24 h to clear their contents from oil droplets and examined under an olympus bx 50 compound microscope equipped with nomarski differential interference contrast optics. microphotographs were recorded on a sony 3ccd color video camera coupled to the microscope. terminology of spore structure is that suggested by stürmer and morton (1997) and walker (1983). spore color was examined under a dissecting microscope on fresh specimens immersed in water. color names are from kornerup and wanscher (1983). nomenclature of plants is after mirek et al. (http://info.botany.pl/czek/check. htm), and that of fungi and the authors of fungal names are those presented at the index fungorum website http://www.indexfungorum.org/authorsoffungalnames. htm. voucher specimens were mounted in pvlg and a mixture of pvlg and melzer’s reagent (1:1, v/v) on slides and deposited in the department of plant protection (dpp), west pomeranian university of technology, szczecin, poland. dna extraction, polymerase chain reaction and dna sequencing. crude dna was isolated from small spore clusters crushed with a needle in ultra clean water on sterile microscope slides under a dissecting microscope. amplification, cloning and sequencing were carried out as described in błaszkowski et al. (2012a). the partial lsu was amplified using the nested pcr with the primer pairs its3-28g2 (da silva et al. 2006; white et al. 1990) and lr1-28g2 (da silva et al. 2006; van tuinen et al. 1998), and the partial nrssu segment was amplified with the primers aml1 and aml2 (lee et al. 2008). pcr was performed with dreamtag green pcr master mix (2x) (thermo scientific, germany). the pcr conditions for lsu were as those described by oehl et al. (2011d), and those for ssu were: initial 5 min denaturation at 94o c followed by 30 cycles of 45 s denaturation at 94o c, 45 s annealing at 58o c, 45 s elongation at 72o c and final 5 min elongation at 72o c. the subsequent work with amplicons was carried out as described in błaszkowski et al. (2012b). sequence alignment and phylogenetic analyses. phylogenetic analyses were performed separately with lsu and ssu sequences. to determine the generic affiliation of the cuban fungus we performed pilot phylogenetic analyses of all its lsu and ssu sequences we obtained with those representing all recognized genera of the glomeromycota with glomoid spores available in genbank. the final data sets comprised all our sequences of the putative s. deserticola, all published sequences of s. deserticola (only lsu sequences), one sequence each of all other described septoglomus spp. (lsu and ssu), except the sequence af145741 (da silva et al. septoglomus deserticola emended and new combinations 93 2006), and sequences of other species with glomoid spores mainly deriving from the former glomus group a (schwarzott et al. 2001). the sequence af145741 probably represents funneliformis coronatus (giovann.) c. walker & a. schüßler (da silva, pers. comm.) whose small-spored isolates closely resemble s. constrictum spores (błaszkowski, pers. observ.). the lsu and ssu sequences were aligned with clustal w (thompson et al. 1994) with default parameters. maximum likelihood (ml) and bayesian (bi) analyses were performed with phyml (guindon and gascuel 2003) and mrbayes 3.1 (huelsenbeck, ronquist 2001; ronquist, huelsenbeck 2003), respectively. before the analyses the best-fit substitution models for the alignments were estimated by the akaike information criterion (aic) using topali v. 2.5 (milne et al. 2004). pacispora scintillans (s.l. rose & trappe) sieverd. & oehl ex c. walker, vestberg & a. schüßler was outgroup in all analyses. in the ml analyses of lsu and ssu sequences the model employed was trn + g, and in the bi analyses of both types of sequences we applied gtr+g and hky+g, respectively. in the ml analysis the transition/transversion ratio for dna models and the gamma distribution parameter were estimated. six substitution rate categories were set. topology and branch lengths and rate parameters were optimized. support of branches in the ml analysis was estimated in a bootstrap analysis with 1000 replicates. in the bi analyses the markov chain was run for 5000000 generations, sampling in every 500 steps, and with a burn-in at 3000. the details of the analyses are available on request. phylogenetic trees were visualized and edited in mega5 (tamura et al. 2011). results molecular analyses. maximum likelihood and bi analyses of lsu sequences generated trees of identical topologies. all sequences of the cuban fungus clustered in a monophyletic group with published sequences of s. deserticola, whose a sister clade comprised all the other known septoglomus spp. except for s. titans (fig. 1). the s. titans sequence formed a separate branch at the base of the septoglomus clade. both the clade with s. deserticola sequences and the other monophyletic groups of the septoglomus clade were well supported [bootstrap supports (bs) of all clades of > 90%, except for one clade of 69%; posterior probabilities (pp) of all clades of 1, except for one clade of 0.89]. the closest molecular relatives of the cuban s. deserticola were s. fuscum błaszk. et al. and s. xanthium błaszk. et al. the topologies of trees obtained following ml and bi analyses of ssu sequences also were identical. the ssu sequences of the cuban fungus grouped in a clade sister to that with sequences of s. africanum, s. constrictum, s. fuscum and s. xanthium (fig. 2). the recently described s. furcatum (blaszkowski et al. 2012) represented a separate linage positioned at the base of the septoglomus clade. the clade with the cuban s. deserticola sequences received very strong supports (bs = 100%, pp = 1), and the supports of the other groups of the septoglomus clade were moderate to high. similarly as in the lsu tree, the cuban s. deserticola was most closely related to s. fuscum and s. xanthium (fig. 2). unfortunately, we could not determine the similarity of the ssu sequences of the cuban s. deserticola with other ssu sequences of this species because of their unavailability in public databases. 94 j. błaszkowski and g. chwat fig. 1. maximum likelihood (ml) tree inferred from lsu nrdna sequences with pacispora scintillans as outgroup. genbank accession numbers of the sequences are in parentheses. ml bootstrap values ≥50% and the bayesian posterior probabilities ≥0.50 are shown near the branches, respectively. sequences of the cuban s. deserticola are in boldface. * = sensu oehl et al. 2011a, ** = sensu schüßler and walker 2010. bar indicates 0.05 expected change per site per branch. septoglomus deserticola emended and new combinations 95 fig. 2. maximum likelihood (ml) tree inferred from ssu nrdna sequences with pacispora scintillans as outgroup. genbank accession numbers of the sequences are in parentheses. ml bootstrap values ≥50% and the bayesian posterior probabilities ≥0.50 are shown near the branches, respectively. sequences of the cuban s. deserticola are in boldface. branch with the corymbiglomus corymbiforme sequence is shortened by 50%. * = sensu oehl et al. 2011a, ** = sensu schüßler and walker 2010. bar indicates 0.02 expected change per site per branch. 96 j. błaszkowski and g. chwat taxonomy septoglomus deserticola (trappe, bloss et j.a. menge) g.a. silva, oehl et sieverd. mycotaxon 116: 106. 2011. figs 3-10 ≡glomus deserticola trappe, bloss & j.a. menge. mycotaxon 20: 123. 1984. spores arise in soil singly or in loose clusters lacking a peridium (figs 3, 7). spores deep yellow (4a8) to light brown (6d8), globose to subglobose, (19–)82(–135) μm diam., sometimes ovoid to pear-shaped, 59–71 × 72–155 μm, with one subtending hypha (figs 3–9). spore wall composed of two layers (figs 4, 5, 7–9). layer 1, forming the spore surface, evanescent, hyaline, (0.5–)1.3(–2.3) μm thick, frequently completely sloughed in mature spores (figs 4, 5, 7–9). layer 2 laminate, smooth, deep yellow (4a8) to light brown (6d8), (1.8–)2.5(–3.8) μm thick, frequently thickened at the spore base to form a collar (figs 4, 5, 7–9). layers 1 and 2 do not stain in melzer’s reagent. subtending hypha deep yellow (4a8) to light brown (6d8), straight or curved, flared, funnel-shaped, rarely constricted at the spore base, (6.3–)8.1–13.3(– 16.8) μm wide at the spore base (figs 5, 7). wall of subtending hypha deep yellow (4a8) to light brown (6d8), composed of two layers continuous with spore wall layers 1 and 2 (figs 5, 7). layer 1 (0.8–)1.0(–1.3) μm thick, layer 2 (2.5–)3.2(–3.8) μm thick; the outer and inner surfaces of layer 2 frequently with side thickenings (figs 5, 7). pore open, (0.8–)3.1–6.5(–10.8) μm wide at the spore base (figs 5, 7). hyphae of clusters deep yellow (4a8) to light brown (6d8), straight or branched, (8.0–)11.3(– 16.8) μm wide, with a 2-layered wall: a hyaline, evanescent, (0.8–)1.5(–2.5) μm thick, when intact, outer layer and a deep yellow (4a8) to light brown (6d8), permanent, (2.0–)2.6(–2.8) μm thick inner layer (figs 3, 5, 6, 9, 10). germination unknown. mycorrhizae. in the field, associated with roots of an unrecognized grass. in addition, lived in symbiosis with parthenium argentatum a. gray, p. incanum kunth, simmondsia chinensis (link) c.k. schneid. (trappe et al. 1984) and uniola paniculata l. (sylvia 1986; sylvia & will 1988). the characters of mycorrhizae from single-species culture of s. deserticola remain unknown. etymology. latin, deserticola (desert dweller), referring to the sandy desert soils in which the fungus was originally found (trappe et al. 1984). specimens examined. poland. szczecin, błaszkowski j., 3144–3146 (dpp), prepared from spores obtained from beg. szczecin, under pot-cultured p. lanceolata, 18 dec. 2012, błaszkowski j., 3353–3357 (dpp), originally coming from the hicacos peninsula, cuba. distribution and habitat. found associated with roots of an unrecognized grass colonizing maritime sand dunes of the hicacos peninsula, cuba. originally described from spores isolated from sandy desert soils of arizona, southern california and texas (u.s.a.; trappe et al. 1984). also recorded in maritime dunes of florida (sylvia 1986; sylvia, will 1988), as well as in other habitats of the u.s.a. (paulitz, menge 1986; bloss, walker 1987; augé 1989), spain (arines, vilarino 1991) and india (ragupathy, mahadevan 1993). probably many times mistakenly identified as rhizophagus fasciculatus (thaxt.) c. walker & a. schüssler (formerly g. fasciculatum (thaxter) gerd. et trappe (trappe et al. 1984; walker, koske 1987), one of the most frequently reported amf from septoglomus deserticola emended and new combinations 97 figs 3–6. septoglomus deserticola from beg. 3. intact spores in a loose cluster. 4. spore wall layers (swl) 1 and 2. 5. spore wall layers (swl) 1 and 2 and subtending hyphal wall layers (shwl) 1 and 2. 6. thick-walled hypha of a cluster and a spore. figs. 7–10. septoglomus deserticola from cuba. 7. spore wall layer (swl) 2 and subtending hyphal wall layer (shwl) 2 of intact spore; swl1 and shwl1 are completely sloughed. 8. spore wall layers (swl) 1 and 2 and subtending hyphal wall layers (shwl) 1 and 2; shwl1 is highly deteriorated. 9. spore wall layers (swl) 1 and 2 and a 2-layered hypha of a cluster. 10. loose hyphae of a cluster. figs. 3, 5–7, 10. spores in pvlg. figs. 4, 8, 9. spores in pvlg+melzer’s reagent. figs 3–10, differential interference microscopy. bars: fig. 3 = 20 μm, figs. 4–10 = 10 μm. 98 j. błaszkowski and g. chwat soil surveys and most often cited as used in studies of plant growth responses (walker 1985). the fungus reported many times under the epithet s. deserticola (błaszkowski 2012) from different regions of poland probably is its close undescribed relative. spores of the fungus are lighter [pale yellow (3a3) to orange (6a6)] and their spore wall layer 1 stains in melzer’s reagent (błaszkowski 1990; vs. no reaction in s. deserticola). notes. morphologically, s. deserticola is most distinguished by its dark-coloured and relatively small spores usually formed in loose clusters (figs 3–9). phylogenetically and morphologically s. deserticola is closest to s. fuscum (figs 1, 2), a species recently described from a material coming from maritime sand dunes located near strand, ca. 50 km southeast of cape town, south africa (błaszkowski et al. 2012). both species produce dark-coloured, relatively small spores, usually in loose clusters and have a 2-layered spore wall (trappe et al. 1984; błaszkowski 2012; błaszkowski et al. 2012). however, the mean diameter of globose s. deserticola spores is almost 2-fold higher, their spore wall layer 1 is short-lived and usually completely sloughed in mature spores (fig. 7; vs. semi-persistent, rarely partly deteriorated in mature and older spores in s. fuscum) and hyaline (figs 4, 5, 8, 9); vs. coloured, rarely hyaline), and spore wall layer 2 is much thinner. in addition, the spore subtending hypha of s. deserticola is much wider and has much thicker walls and a wider pore. septoglomus xanthium, another species close in phylogeny to s. deserticola (figs 1, 2), also forms spores almost indistinguishable from those of the latter fungus when they are intact and seen under a dissecting microscope. the two species mainly separate the number of spore wall layers (three vs. two in s. deserticola; figs. 4, 5, 8, 9) and the phenotypic features of spore wall layer 1, forming the spore surface (semi-permanent and coloured in mature spores vs. hyaline and usually completely sloughed at maturity; figs. 4, 5, 7–0; trappe et al. 1984; błaszkowski et al. 2004; błaszkowski 2012). in addition, s. xanthium spores are clearly smaller [(23–)50–70) µm diam when globose vs. (19–)82(– 135) µm diam when globose], have a much thinner structural laminate spore wall layer and a narrower subtending hypha with thinner walls and a narrower pore. of other known species of the glomeromycota, s. deserticola may be confused with small-spored isolates of funneliformis coronatus (giovann.) c. walker & a. schüssler, s. constrictum and s. furcatum błaszk., chwat & kovács, ryszka due to their similarly coloured spores (błaszkowski 2012; błaszkowski et al. 2012). however, spores of the three latter species usually are much larger and arise only singly (vs. singly and in clusters in s. deserticola; figs 3–9). other differences between the four species reside in their spore wall structure, phenotypic and histochemical characters of spore wall layers, spore subtending hyphal features and, most importantly, in their phylogenies (figs 1, 2). emendations and new combinations diversisporaceae c. walker & a. schüßler, emend. błaszk. et chwat forming spores blastically at the top of a sporogenous hypha (glomus-like spores), laterally on the neck of a sporiferous saccule (acaulospora-like spores) or inside the neck of a sporiferous saccule (entrophospora-like spores). acaulospora-like septoglomus deserticola emended and new combinations 99 spores and entrophospora-like spores as otosporoid and tricisporoid spores, respectively, sensu oehl et al. (2011e). glomus-like spores without a hyphal mantle (diversisporoid spores sensu oehl et al. 2011e) or covered individually with a hyphal mantle consisting of non-branched or branched hyphae. mantled spores occurring singly or in clusters with two to three spores grouped by interwoven hyphae of their hyphal mantle or in clusters with two to <20 spores formed by spores arisen at the top of sporogenous hyphae dichotomously branched from a parent hypha continuous with an extraradical mycorrhizal hypha. spores pigmented, with a 1–3-layered spore wall. subtending hypha straight or recurved, cylindrical to funnel-shaped or constricted. subtending hyphal wall continuous with and coloured similarly to the spore wall. pore open or occluded by a septum continuous with the innermost laminae of the structural spore wall, by thickening of the structural spore wall or by spore wall layer 3. type genus: diversispora c. walker et a. schüssler emend. g.a. silva, oehl et sieverd. mycotaxon 116: 108. 2011c. other genera: corymbiglomus błaszk. et chwat. glomeromycota 272. 2012. otospora oehl, palenz. et n. ferrol. mycologia 100: 297. 2008 redeckera c. walker et a. schüssler, emend. oehl, g.a. silva &sieverd. mycotaxon 116: 110. 2011c. tricispora oehl, sieverd., g.a. silva et palenz. mycotaxon 117: 310. 2011e. corymbiglomus błaszk. et chwat, emend. forming glomus-like spores individually covered with a hyphal mantle consisting of non-branched or branched hyphae with or without terminal vesiculate swellings. spores occurring singly or in clusters. clusters with two to three spores grouped by interwoven hyphae of their hyphal mantle or with two to <20 spores arisen at the top of sporogenous hyphae dichotomously branched from a parent hypha continuous with an extraradical mycorrhizal hypha. spores pigmented, with a 1–3-layered spore wall. subtending hypha straight or recurved, cylindrical to funnel-shaped or constricted. subtending hyphal wall continuous with and coloured similarly to the spore wall. pore open or occluded by a septum continuous with the innermost laminae of the structural spore wall, by thickening of the structural spore wall or by spore wall layer 3. type species: corymbiglomus corymbiforme (błaszk.) błaszk. et chwat. glomeromycota 274. 2012. other species: corymbiglomus globiferum (koske et walker) błaszk et chwat, comb. nov. ≡glomus globiferum koske et walker. mycotaxon 26: 133. 1986. corymbiglomus tortuosum (n.c. schenck et g.s. sm.) błaszk. et chwat, comb. nov. ≡glomus tortuosum n.c. schenck et g.s. sm. mycologia 74: 83. 1982. notes. the genus corymbiglomus was originally erected based on phylogenetic analyses of lsu sequences of g. corymbiforme błaszk. (błaszkowski 2012). results of phylogenetic analyses of lsu and ssu sequences presented here confirmed those 100 j. błaszkowski and g. chwat of earlier studies (błaszkowski 2012) and clearly revealed that species grouping with c. corymbiforme in a clade with lsu sequences also are g. globiferum and g. tortuosum (figs 1, 2). similarly as in the phylogenetic analyses of lsu sequences, those of ssu sequences placed c. corymbiforme in a position sister to the clade with known diversispora spp. unfortunately, we could not determine the c. corymbiglomus x c. globiferum x c. tortuosum relationship found in the analyses of lsu sequences, because there are no ssu sequences of the latter two species in available databases. morphologically, the species link that their spores are covered with a hyphal mantle and they usually occur in clusters. acknowledgements this study was supported in part by the national centre of science, grants no. n n304 061739 and 2012/05/b/nz8/00498. ananonymous reviewers 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(eds). pcr protocols: a guide to methods and amplifications: 315–322. academic press, san diego, california. uzupełnione diagnozy dwóch taksonów – septoglomus deserticola i diversisporaceae oraz nowe kombinacje w obrębie rodzajów glomus i corymbiglomus streszczenie przedstawiono uzupełnioną definicję morfologii zarodników septoglomus deserticola, arbuskularnego grzyba mikoryzowego z gromady glomeromycota, na podstawie oryginalnego opisu tego gatunku, jego jedynej innej definicji opublikowanej niedawno i zarodników wyhodowanych w kulturach wazonowych zainokulowanych glebą ryzosfrową i fragmentami korzeni nierozpoznanej trawy kolonizującej piaszczyste wydmy nadmorskie półwyspu hicacos, kuba. septoglomus deserticola emended and new combinations 103 analizy filogenetyczne sekwencji dużej podjednostki (lsu) jądrowego rdna kubańskiego grzyba potwierdziły jego powinowactwo z s. deserticola zdeponowanym w międzynarodowym banku glomeromycota (beg) i wykazały, że jego najbliższymi krewniakami są s. fuscum i s. xanthium. analizy filogenetyczne sekwencji małej podjednostki (ssu) jądrowego rdna potwierdziły pokrewieństwo trzech taksonów: grzyb kubański x s. fuscum x s. xanthium ujawnione w analizach sekwencji lsu i przez to zasugerowały, że kubańskie s. deserticola jest właściwym s. deserticola. jednak nie było możliwości dowieść bezpośrednio identyczności kubańskiego s. deserticola z typowym s. deserticola z beg na podstawie sekwencji ssu z powodu braku sekwencji ssu s. deserticola w dostępnych bazach danych. ponadto analizy filogenetyczne sekwencji lsu i ssu potwierdziły unikatowość niedawno utworzonego rodzaju corymbiglomus z gatunkiem typowym c. corymbiforme (wcześniejszym glomus corymbiforme) w rodzinie diversisporaceae i dowiodły, że jego sekwencje lsu grupują się w kladzie z sekwencjami lsu glomus globiferum i g. tortuosum. w konsekwencji dwa ostatnie gatunki zostały przeniesione do rodzaju corymbiglomus i nazwane c. globiferum comb. nov. oraz c. tortuosum comb. nov., a definicje rodziny diversisporaceae i rodzaju diversispora zostały uzupełnione. 2013-06-22t22:34:42+0100 polish botanical society new records of lichens from the polish uplands anna łubek institute of biology, jan kochanowski university, świętokrzyska 15 pl-25-406 kielce, anna.lubek@ujk.kielce.pl łubek a.: new records of lichens from the polish uplands. acta mycol. 44 (2): 275–282, 2009. five lichen species: bacidia pycnidiata, fellhanera gyrophorica, leucocarpia biatorella, parmotrema perlatum and punctelia ulophylla, are presented. bacidia pycnidiata, leucocarpia biatorella, punctelia ulophylla are new to central poland. bacidia pycnidiata is reported from poland for the third time. key words: rare lichens, new sites, central poland introduction studies on lichens in the świętokrzyskie mts. (kielce and the chęcińsko-kielecki landscape park) and adjoining sites (the oleszno nature reserve in the przedborski landscape park) (fig. 1) have been conducted in recent years. a few interesting species of lichens that are very rare in poland were found. those are: bacidia pycnidiata, fellhanera gyrophorica, leucocarpia biatorella, parmotrema perlatum and punctelia ulophylla. two of them, bacidia pycnidiata and fellhanera gyrophorica, have lately been described to science. intensive lichenological research carried out in different regions of poland, including the świętokrzyskie mts., provides new data on the occurrence of these lichens as each new and verified site is very important for the knowledge on habitat preferences and the material distribution of the species in poland. the main goal of this work is to provide the most up-to-date information on the above species. acta mycologica vol. 44 (2): 275–282 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity new records of lichens 277 distribution. europe: belgium, the czech republic, estonia (czarnota, coppins 2006; suija et al. 2007; diederich et al. 2009). in poland, b. pycnidiata is known from mountainous regions: the western beskidy mts. and the pogórze środkowobeskidzkie foothills (czarnota, coppins 2006). specimens examined. atpol grid square ee 60 − przlp, near the oleszno nature reserve, forest section no 79, on bryophytes and on a trunk of fraxinus excelsior, 7 feb. 2008 (ktc 8157). ee 73 − ch-klp, the pasmo zgórskie range, at the edge of the forest near the village of zagórze, on a trunk of quercus sp., 9 aug. 2007 (ktc 7994). note. the species is reported as new to the świętokrzyskie mts. this is the third site in poland. fellhanera gyrophorica sérus., coppins, diederich & scheid., lichenologist 33 (4): 285 (2001). for the description of the species see sérusiaux et al. (2001) and sparrius (2002). f. gyrophorica is usually sterile and produces only large brown pycnidia reacting c+ red. apothecia were found only once in the material collected from the puszcza białowieska forest (sparrius 2002). the specimens from the przedborski landscape park have pycnidia only. f. gyrophorica grows on tree bark or invades corticolous mosses and liverworts in old-growth forests. distribution. europe: austria, luxembourg, switzerland, estonia, lithuania, belarus, ukraine and slovakia (sérusiaux et al. 2001; motiejūnaitė, prigodina-lukošienė 2002; motiejūnaitė et al. 2003; golubkov, kukwa 2006; pišút et al. 2007). f. gyrophorica is very rare in poland. it is known from northern poland: gdańsk pomerania (kukwa 2006/2007), the wysoczyzna elbląska high plain (szymczyk, kukwa 2008) and north-east poland: the puszcza białowieska forest (czyżewska et al. 2001; sparrius 2002; 2003), the puszcza borecka forest (sérusiaux et al. 2001), the budzisk nature reserve in the puszcza knyszyńska forest (czyżewska et al. 2002), the biebrza national park (czyżewska et al. 2005). in central poland it has been reported from the puszcza kozienicka forest (cieśliński 2007) and the spała nature reserve in the puszcza pilicka forest (motiejūnaitė, czyżewska 2008). specimens examined. ee 60 − przlp, near the oleszno nature reserve, forest section no 73, on a trunk of fraxinus excelsior, 11 mar. 2008 (ktc 8018), the oleszno nature reserve, forest section no 54, on a trunk of quercus sp., 25 may 2008 (ktc 8025). leucocarpia biatorella (arnold) vĕzda, herzogia 1: 192 (1969). syn. microglaena biatorella arnold, microthelia biatorella (arnold) dalla torre & sarnth. for the description of the species see purvis et al. (1992). l. biatorella grows on soil and calcareous stones associated with mosses. it is similar to chromatochlamys muscorum which has larger ascospores 2-4 per ascus, and polyblastia gelatinosa which has black perithecia and grows on bryophytes. the specimen from the chęcińsko-kielecki landscape park has a pale granular-verrucose thallus and immersed perithecia with a flat yellow-pinkish ostiolar region. ascospores are eight per ascus, hyaline and muriform, 28–36 × 11–15 µm. distribution. north america: continental united states and canada (esslinger 2008); europe: great britain and ireland, sweden and finland (santesson 1993; vitikainen et al. 1997; coppins 2002), asia: mongolia (biazrov 2009). l. biatorella has so far been reported from southern poland – the western carpathians mts. (olech 1999 and literature cited in: flakus 2007), near the town 278 a. łubek of olkusz (kiszka, kościelniak 2006), the śnieżnik massif and the bialskie mts. (szczepańska 2008). specimen examined. ee 83 − ch-klp, the pasmo zelejowskie range, n slope of wiśniowa mt., on stones in shady and wet places, 16 aug. 2007 (ktc 7995). note. species reported as new to central poland and the świętokrzyskie mts. parmotrema perlatum (huds.) m. choisy, bull. mens. soc. linn. lyon 21: 174 (1952). syn. p. chinense (osbeck) hale & ahti, parmelia perlata (huds.) ach., p. trichotera hue. for the description of the species see purvis et al. (1992). p. perlatum grows on well-lit, neutral to somewhat acid-barked, broad-leaved trees and also frequently on siliceous rocks and walls. the species is sensitive to the mean so2 level in the atmosphere. the species grows only in the best parts of old-growth forests according to motyka (1960). the specimen collected in the przedborski landscape park has a very well developed thallus with marginal soralia and cilia. it was observed in small groups only on one tree in a sunny place. distribution. europe: great britain and ireland, denmark, norway, germany, the czech republic, romania (santesson 1993; vězda, liška 1999; scholz 2000; coppins 2002; søchting, alstrup 2007; ciurchea 2009); asia: china (checklist of lichens and lichenicolous fungi of yunnan), japan (kurokawa 2003), thailand (wolseley et al. 2002), turkey (checklist of lichens and lichenicolous fungi of turkey); new zealand (galloway 2007); north america: continental united states and canada (tucker, ryan 2006; esslinger 2008); south america: argentina (checklist of lichens and lichenicolous fungi of argentina); africa: algeria (checklist of lichens and lichenicolous fungi of algeria), tanzania (checklist of lichens and lichenicolous fungi of tanzania); australia (elix, mccarthy 2008). it has been reported in poland from the wyżyna lubelska upland, the kotlina sandomierska basin, the gorce mts., the beskid sądecki mts. and the bieszczady mts. in the 19th and the 20th-century literature (motyka 1960; sulma, fałtynowicz 1988; bielczyk 1997; see also fałtynowicz 2003). lately it has only been reported from the polish eastern carpathians (kiszka, kościelniak 1998; kościelniak 2008). specimen examined. ee 60 − przlp, near the oleszno nature reserve, forest section no 73, on a trunk of fraxinus excelsior, 11 mar. 2008, det. m. kukwa (ktc 8004). note. p. perlatum is an endangered species in poland (cr) (cieśliński et al. 2003). punctelia ulophylla (ach.) van herk & aptroot, lichenologist 32 (3): 239 (2000). syn. p. subrudecta var. ulophylla (ach.) harm. for the description of the species see van herk and aptroot (2000). p. ulophylla is known only to be corticolous. it grows on a wide variety of trees. p. ulophylla can be mistaken for p. subrudecta. extreme margins of the thallus of p. ulophylla are dull brownish and pruinose. secondary lobes have dense marginal soredia. extreme margins of the thallus of p. subrudecta are dark brown, glossy and not pruinose, marginal soredia are absent and only soredia laminal are present (van herk, aptroot 2000). thalli of punctelia ulophylla from the przedborski landscape park were recorded abundantly on thick branches. distribution. europe: great britain, belgium, france, the netherlands, germany, poland, slovakia, switzerland (van herk, aptroot 2000; coppins 2002), northern america (tucker, ryan 2006). new records of lichens 279 the species has been reported in poland from the sudetes mts., the western bieszczady mts. (sulma, fałtynowicz 1988; see also fałtynowicz 2003 and van herk, aptroot 2000). specimens examined. ee 60 − przlp, near the oleszno nature reserve, forest section no 79, in the crown of fraxinus excelsior, 7 feb. 2008, det. m. kukwa (ktc 8003, 8006), forest section no 73, in the crown of fraxinus excelsior, 7 feb., 11 mar. 2008, det. m. kukwa (ktc 8086, 8005) and the oleszno nature reserve, forest section no 60, in the crown of fraxinus excelsior, 25 may 2008 (ktc 8054). note. species new to central poland. conclusions 1. of the lichens collected, fellhanera gyrophorica is especially noteworthy. it is connected with old-growth forests and its richest occurrence was observed in big forest complexes such as the puszcza białowieska forest, the puszcza borecka forest, the puszcza knyszyńska forest, the puszcza pilicka forest or the puszcza kozienicka forest. the occurrence of f. gyrophorica in the oleszno nature reserve and in its vicinity could result from good habitat conditions, especially a high level of air humidity. a natural mixed family tree-stand, permanently waterlogged in some places, with a considerable participation of alnus glutinosa and fraxinus excelsior, is protected within the oleszno nature reserve. less accessible areas are unaffected by human activity and are also a refuge to other interesting lichen species, such as parmotrema perlatum. 2. it is very interesting that leucocarpia biatorella, which grows mostly in the high part of mountains, has been recorded from the świętokrzyskie mts. the occurrence of the species here may be connected with the presence of metals in the substrate, such as zinc, lead, iron and limestone, or its tolerance to heavy metals in the substrate is high (cf. kiszka, kościelniak 2006). limestone, marble from chęciny and sparite were extracted in the pasmo zalejowskie range in the past and copper, lead and silver ores were found in the neighbouring pasmo chęcińskie range where the species was observed. the location of l. biatorella may be situated at the site of previous vein extraction. this is supported by the presence of many wells of different depth in the ground around which numerous rock blocks covered with bryophytes occur. beechwood that grows at the site makes it shaded and quite dump. it is another anthropogenic site of l. biatorella in poland apart from the wyżyna śląska upland. lichenological research conducted in other parts of the świętokrzyskie mts. where limestone occurs did not reveal the presence of this species. further research is required to complete ecological requirements of the species and to describe the species accurately. acknowledgements. i would like to thank prof. stanisław cieśliński (jan kochanowski university, kielce), dr. hab. paweł czarnota (scientific laboratory, the gorce national park) and dr. martin kukwa (university of gdańsk) for determining or revising the determination of some taxa. thanks are also due the anonymous reviewer for valuable comments and 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in california. constancea 84. university and jepson herbaria, california lichen catalog. available at http:// ucjeps.berkeley.edu/constancea/84/index.html [date of exploration: 2 june 2009]. 282 a. łubek vězda a., liška j. 1999. katalog lišejníků české republiky. institute of botany, academy of sciences of the czech republic, průhonice, 283 pp. vitikainen o., ahti t., kuusinen m., lommi s., ulvinen t. 1997. checklist of lichens and allied fungi of finland. norrlinia 6: 1–123. wolseley p. a., aguirre-hudson b., mccarthy p. m. 2002. catalogue of the lichens of thailand. bull. nat. hist. museum london (bot.) 32 (1): 13–59. nowe notowania porostów z polskich wyżyn streszczenie w pracy przedstawiono pięć gatunków porostów (bacidia pycnidiata, fellhanera gyrophorica, leucocarpia biatorella, parmotrema perlatum i punctelia ulophylla), które występowały na sześciu nowych stanowiskach zlokalizowanych w polsce środkowej. badane materiały zdeponowane są w zielniku ktc. wśród stwierdzonych gatunków, bacidia pycnidiata, leucocarpia biatorella i punctelia ulophylla są porostami zanotowanymi w polsce środkowej po raz pierwszy. jednocześnie stanowisko bacidia pycnidiata jest trzecim stwierdzonym stanowiskiem w polsce. gatunkami nowymi dla gór świętokrzyskich są bacidia pycnidiata i leucocarpia biatorella. 2014-01-01t11:49:56+0100 polish botanical society glomus eburneum and scutellospora fulgida, species of arbuscular mycorrhizal fungi (glomeromycota) new for europe janusz błaszkowski and beata czerniawska department of plant pathology, university of agriculture in szczecin słowackiego 17, pl-71-434 szczecin, jblaszkowski@agro.ar.szczecin.pl błaszkowski j., czerniawska b.: glomus eburneum and scutellospora fulgida, species of arbuscular mycorrhizal fungi (glomeromycota) new for europe. acta mycol. 43 (1): 57–65, 2008. morphological characters of spores and mycorrhizae of glomus eburneum and spores of scutellospora fulgida, arbuscular mycorrhizal fungi of the phylum glomeromycota, are described and illustrated. additionally, the known distribution of these species in both poland and other regions of the world is presented. both species were not earlier reported from europe. key words: glomus eburneum, glomeromycota, scutellospora fulgida, occurrence introduction examination of pot trap cultures with mixtures of rhizosphere soils and roots collected in poland and other regions of the world revealed spores of glomus eburneum l.j. kenn., j.c. stutz et j.b. morton and scutellospora fulgida koske et c. walker, arbuscular mycorrhizal fungi of the phylum glomeromycota. both species were not earlier reported from europe. the aims of this paper are to describe and illustrate these species and present their distribution in both poland and the world. materials and methods establishment and growth of trap and one-species cultures, extraction of spores, and staining of mycorrhizae. spores examined in this study came from both pot trap and one-species cultures. trap cultures were established to obtain a great number of living spores of different developmental stages and to initiate sporulation of species that were present but not sporulating in the field collections (stutz, morton 1996). the method used to establish trap cultures, their growing conditions, and the method of spore extraction were previously described (błaszkowski et al. 2004, 2006). acta mycologica vol. 43 (1): 57–65 2008 58 j. błaszkowski and b. czerniawska one-species cultures were also generally established and grown as given in błaszkowski et al. (2004), with two exceptions. first, instead of marine sand, their growing medium was an autoclaved commercially available coarse-grained sand (grains 1.0-10.0 mm diam. 80.50%; grains 0.1-1.0 mm diam. 17.28%; grains < 0.1 mm diam. 2.22%) mixed (5:1, v/v) with clinopthilolite (zeocem, bystré, slovakia) of grains 2.5-5 mm. clinopthilolite is a crystaline hydrated alumosilicate of alkali metals and alkaline earth metals having, e.g., a high ion exchange capability and selectivity, as well as a reversible hydration and dehydration. ph of the sandclinopthilolite mixture was 7.3. second, the cultures were kept in transparent plastic bags, 15 cm wide and 22 cm high as suggested by walker and vestberg (1994), rather than open pot cultures (gilmore 1968). to prevent contamination of the cultures with other amf but still to allow exchange of gases, we left an opening, ca. 1 cm wide, in the centre of the upper part of each bag, while the edges on both sides were closed with small plastic clips. the cultures were watered with tap water once a weak, harvested after five months when spores were extracted for study. to reveal mycorrhizae, root fragments located ca. 1-5 cm below the upper level of the growing medium were cut off with a scalpel. the host plant used in both trap and one-species cultures was plantago lanceolata l. microscopy survey. morphological properties of spores and their wall structure were determined based on examinations of at least 100 spores mounted in polyvinyl alcohol/lactic acid/glycerol (pvlg; omar, bollan and heather 1979) and a mixture of pvlg and melzer’s reagent (1:1, v/v). spores at all developmental stages were crushed to varying degrees by applying pressure to the cover slip and then stored at 65oc for 24 h to clear their contents from oil droplets. these were then examined under an olympus bx 50 compound microscope equipped with nomarski differential interference contrast optics. microphotographs were recorded on a sony 3cdd color video camera coupled to the microscope. terminology of spore structure is that suggested by stürmer and morton (1997) and walker (1983). spore colour was examined under a dissecting microscope on fresh specimens immersed in water. colour names are from kornerup and wanscher (1983). nomenclature of fungi and plants is that of walker and trappe (1993) and mirek et al. (1995), respectively. the authors of the fungal names are those presented at the index fungorum website http://www.indexfungorum.org/authorsoffungalnames.htm. specimens were mounted in pvlg on slides and deposited in the department of plant pathology, university of agriculture, szczecin, poland. colour microphotographs of spores and mycorrhizae of the fungi presented here can be viewed at the url http://www.agro.ar.szczecin.pl/~jblaszkowski/. descriptions of the species glomus eburneum l.j. kenn., j.c. stutz et j.b. morton sporocarps unknown. spores occur singly in the soil (fig. 1); origin blastically at the tip of hyphae continuous with extraradical mycorrhizal hyphae. spores yellowish white (4a2) to butter yellow (4a5); globose to subglobose; (92-)108(-146) μm diam; or ovoid; 104-115 x 123-139 μm; with one subtending hypha (figs 1, 2, 5 and 6). subcellular structure of spores consists of a spore wall composed of two tightly adherent layers (layers 1 and 2; figs 2-5). layer 1, forming the spore surface, semi glomus eburneum and scutellospora fulgida 59 permanent, semi-flexible, hyaline, (0.7-)1.0(-1.2) μm thick, frequently intact or only slightly deteriorated in mature spores, sometimes associated with granular soil debris. layer 2 laminate, semi-flexible, smooth, yellowish white (4a2) to butter yellow (4a5), (2.5-)3.5(-4.4) μm thick. none of the two layers stains in melzer’s reagent. subtending hypha yellowish white (4a2) to butter yellow (4a5); straight or recurved; cylindrical or slightly flared, sometimes slightly constricted; (7.8-)9.9(-14.0) μm wide at the spore base (figs 1, 5 and 6). wall of subtending hypha yellowish white (4a2) to butter yellow (4a5); (1.0-)1.6(-2.7) μm thick at the spore base; composed of two layers continuous with spore wall layers 1 and 2 (fig. 5). pore 3.8-5.1 μm diam. open or occluded by a straight or recurved septum, (1.0-)1.4(-2.0) μm thick, continuous with the innermost laminae of the laminate spore wall layer 2, positioned up to 11.3 μm below the spore base (figs 5 and 6). germination. not observed. mycorrhizal associations. the presence of mycorrhizae in field-collected root samples was not determined. in one-species cultures with p. lanceolata as the host plant, mycorrhizae of g. eburneum comprised arbuscules, as well as intraand extraradical hyphae. arbuscules consisted of short trunks branched from parent hyphae and numerous branches with very fine tips (figs 7 and 8). their distribution along root fragments ranged from uniform to patchy, depending on the root fragment examined. intraradical hyphae grew along root axis, were (2.0-)4.9(-8.3) μm wide and sometimes formed yor h-shaped branches and coils (figs 7 and 8). the coils were ellipsoid, 10.523.3 x 36.8-57.4 μm, when observed in a plane view. extraradical hyphae were (2.7-) 3.1(3.4) μm wide and occurred very rarely and in low abundances. in 0.1% trypan blue, arbuscules stained violet white (15a2) to light lilac (16a5), intraradical hyphae violet white (16a2) to pale violet (16a3), coils lilac (16a5) to royal purple (16d8), and extraradical hyphae violet white (16a2) or remained unstained. phylogenetic position. kennedy, stutz and morton (1999) concluded that the lack of vesicles and the faintly staining other components of mycorrhizae of g. eburneum are untypical of most species of the genus glomus, but are characteristic for, e. g., archaeospora trappei (r.n. ames et linderman) j.b. morton et d. redecker emend. spain, appendicispora gerdemannii (s.l. rose, b.a. daniels et trappe) spain, oehl et sieverd. and diversispora spurca (c.m. pfeiff., c. walker et bloss) c. walker et schuessler, of which the former two fungi come from the order archaeosporales c. walker et schuessler and the latter one has recently been accommodated in the family diversisporaceae c. walker and schuessler of the order diversisporales c. walker et schuessler (schüßler, schwarzott and walker 2001; spain, sieverding and oehl 2006; walker and schüßler 2004). gamber and leuchtmann (2007) found the invam isolate az420a of g. eburneum to be in a sister position to g. versiforme (p. karsten) s.m. berch, being phylogenetically most closely related to d. spurca (schwarzott, walker and schüßler 2001). distribution and habitat. in poland, spores of g. eburneum were found in only one trap culture containing a mixture of rhizosphere soil and root fragments taken from under helichrysum areanarium (l.) moench growing in maritime sand dunes of the słowiński national park (54o45’n, 17o26’e) on 26 june 2003. the occurrence of spores of arbuscular fungi in the field sample was not investigated. the species of arbuscular fungi co-occurring with g. eburneum in the trap culture were archaeospora 60 j. błaszkowski and b. czerniawska trappei (r.n. ames et linderman) j.b. morton et d. redecker emend. spain and g. lamellosum dalpé, koske et tews. the holotype of g. eburneum has been selected from spores extracted from the invam culture az420a established from a mixture of rhizosphere soil and root fragments of sporobolus wrightii monro ex scribn. growing along the san pedro river in arizona, u.s.a. (kennedy, stutz and morton 1999). sporobolomyces wrightii is a native grass species found only along rivers and streams of the semiarid regions of south-western north america. additionally, the same scientists and stutz et al. (2000) recorded this fungus among roots of other plants growing in other sites of arizona and mexico, as well as under different plant species colonizing a dune transect in the namibia desert. collections examined. poland: szczecin, under pot-cultured p. lanceolata, 10.02.2005, j. błaszkowski 2682-2696 (dpp). notes. the most distinctive characters of g. eburneum are its light-coloured spores filled with dense, opaque oil substance and the semi-flexible, 2-layered spore wall with the outermost layer nonreactive in melzer’s reagent and usually remaining more or less intact in mature spores (figs 1-6). when observed at a low magnification, spores of g. eburneum are most reminiscent of those of d. spurca, g. albidum c. walker et l.h. rhodes, g. gibbosum błaszk., g. viscosum nicol., and paraglomus occultum (c. walker) j.b. morton et d. redecker. darker-coloured spores of g. eburneum may also be confused with lightpigmented spores of g. claroideum n.c. schenck et s.m. sm. and g. versiforme. except for g. versiforme, examination of spores crushed in pvlg and pvlg mixed with melzer’s reagent under a compound microscope readily separates g. eburneum from all the other species listed above. considering the spore wall structure, as well as the phenotypic and biochemical properties of its components, the fungus most closely related to g. eburneum is d. spurca. although the opinions of the number of layers overlaying the laminate spore wall layer of d. spurca are contradictory (one layer according to kennedy, stutz and morton 1999 and morton 2002 vs. two layers as błaszkowski (2003) stated), the laminate layer and the layer directly overlaying it are almost identical in both their phenotypic and biochemical properties. the main differences between these fungi hide in the persistency of the spore wall layers directly covering the structural laminate layer and the degree of the association of these layers with the laminate layer. consequently, the second property defines the basic differences in the persistency of the subtending hyphae of both fungi. although the outer spore wall layer is relatively long-lived and usually retains as a more or less deteriorated structure in mature spores of g. eburneum (figs 2-5), its spatially corresponding spore wall layer in d. spurca is more persistent and always remains intact in even old spores (błaszkowski 2003; kennedy, stutz and morton 1999). in g. eburneum, the outer spore wall layer always remains tightly adherent to the laminate spore wall layer (figs 2-5), whereas the semi-flexible layer directly covering the laminate spore wall layer of d. spurca easily separates in crushed spores or balloons when immersed in lactic acid-based mountants (błaszkowski 2003; kennedy, stutz and morton 1999). glomus eburneum and scutellospora fulgida 61 in g. eburneum, the structural layer of the subtending hyphal wall (shwl2) continuous with the laminate spore wall layer and being the support of the outer thin wall layer of the subtending hypha gradually thins and expands up to 25 μm below the spore base (fig. 5). in contrast, in d. spurca spores, the laminate spore wall layer abruptly thins and stops to grow at their base and, thereby, it does not create a sufficient support to stabilize the outer subtending hyphal wall layer continuous with the spore wall layer 2 sensu błaszkowski (2003). therefore, almost all crushed spores of d. spurca usually lack the subtending hypha, which detaches along with the outer wall layer of these spores. finally, in contrast to the opaque, frequently yellow-coloured contents of g. eburneum spores, the spore contents of d. spurca consists of transparent oil droplets. compared with g. albidum described to also form a 2-layered spore wall of similar phenotypic characters (walker, rhodes 1981) to those of the wall layers of spores of g. eburneum, spores of the latter species do not react in melzer’s reagent (vs. become pink to orange red in g. albidum; walker, rhodes 1981). two characters of spores of g. gibbosum readily separate this fungus from g. eburneum. first, while spores of the former species occur in the soil singly, in loose aggregates and conglomerations enclosed by a common hyphal mantle (błaszkowski 1997, 2003), the latter fungus produces only single spores (fig. 1). second, the spore wall of g. eburneum comprises only two layers (figs 2-5), and that of g. gibbosum consists of four layers (błaszkowski 2003). the spore wall of g. eburneum lacks the wall layers 2 and 4 of spores of g. gibbosum. spores of g. viscosum are also frequently formed in loose aggregates (morton 2002; vs. only single spores in g. eburneum; fig. 1) and have a more complex wall structure (3-layered) than those of g. eburneum (2-layered; figs 2-5). the spore wall layer of g. viscosum not synthesized by g. eburneum is the permanent, semi-flexible, thin layer positioned between a semi-flexible layer forming the spore surface and a laminate layer, both phenotypically similar to the spore wall layers 1 and 2, respectively, of g. eburneum. as far as the species compared here are concerned, the fungus most diverged morphologically from g. eburneum is paraglomus occultum. these species share only their outermost spore wall layer (morton 2002; morton, redecker 2001). although these layers in both fungi are of the type of impermanent layers, the longevity of the layer in g. eburneum is much higher than in p. occultum, in which it usually highly deteriorates with age to form a granular structure (morton 2002), a phenomenon not found in g. eburneum (figs 2-6). the two other spore wall layers of p. occultum are uniform and much thinner (both <0.5-1.2 μm thick) than the inner laminate spore wall layer of g. eburneum [(2.5-)3.5(-4.4) μm thick]. as mentioned above, darker-coloured spores of g. eburneum may also overlap in colour and appearance with light-coloured spores of g. claroideum and g. versiforme. however, the former two fungi differ fundamentally in the construction of their spore wall, as well as in the phenotypic and biochemical properties of its components. compared with the simple, 2-layered spore wall of g. eburneum (figs 2-6), that of g. claroideum comprises four layers with the innermost layer staining in melzer’s reagent (błaszkowski 2003; morton 2002; stürmer, morton 1997; vs. none of the spore wall layers of g. eburneum reacts in this reagent; fig. 4). the only layers of the spore wall of g. claroideum sharing the phenotypic and biochemical 62 j. błaszkowski and b. czerniawska properties of layers 1 and 2 of the g. eburneum spore wall are its layers 2 and 3, respectively. the distinctive component of the spore wall of g. claroideum is the innermost flexible layer, which is lacking in g. eburneum. light-coloured spores of g. versiforme may be indistinguishable from mature spores of g. eburneum when observed under both a dissecting and a compound microscope. apart from colour, spores of the two species are almost identical in size, as well as in the construction and the phenotypic and biochemical properties of the components of their wall. moreover, mycorrhizae of both species stain faintly in 0.1% trypan blue (figs 7 and 8; błaszkowski, pers. observ.; morton 2002). the only property distinguishing g. eburneum and g. versiforme is the formation of sporocarps by the latter fungus (morton 2002). apart from the morphological differences characterized above, the species compared here also differ in the phylogenetic position within the phylum glomeromycota determined based on results of their molecular analyses. for example, g. claroideum and g. viscosum represent glomus group b in the family glomeraceae piroz. et dalpé of the order glomerales j.b. morton et benny, and par. occultum, originally described as g. occultum c. walker, presently is a member of the family paraglomaceae j.b. morton et d. redecker in the paraglomerales c. walker et schuessler (schüßler, schwarzott and walker 2001). as presented in the section “phylogenetic position”, g. eburneum should be a member of the family diversisporaceae, as gamber and leuchtmann (2007) found. unfortunately, the phylogenetic positions of g. albidum and g. gibbosum are unknown to date. scutellospora fulgida koske et c. walker spores formed singly in the soil; origin blastically at the tip of a bulbous sporogenous cell; yellowish white (4a2) to cream (4a3); globose to subglobose; (165-)229 (-280) µm diam (fig. 9). subcellular structure of spores consists of a spore wall and one inner germination wall (figs 10-13). spore wall composed of two layers (layers 1 and 2; figs 10, 11, 13 and 14). layer 1, forming the spore surface, permanent, smooth, pale yellow (3a3) to light orange (5a5), (1.2-)2.0(-3.2) µm thick, usually tightly adherent to layer 2, sometimes slightly separated from it in vigorously crushed spores. layer 2 laminate, smooth, yellowish white (4a2) to cream (4a3), (7.6-)9.7 (-13.0) µm thick. germination wall comprises two flexible, smooth layers (layers 1 and 2; figs 10-13). layer 1 ca. 0.5 µm thick, adherent to layer 2, but frequently wrinkled in crushed spores and, thereby, giving the inner wall a rugose or blistered appearance and making this layer more visible. layer 2 0.8-1.5 µm thick. in melzer’s reagent, the spore wall layers 1 and 2 stain orange white (5a2) to copper (7c8) and pale yellow (3a3) to reddish orange (7b7), respectively, and both germination wall layers remain nonreactive (figs 11-14). sporogenous cell formed terminally on a sparsely septate hypha continuous with an extraradical mycorrhizal hypha; ovoid to clavate; (38.7-)44.6(-53.2) µm wide; cream (4a3) to light orange (5a5), usually with a hyphal peg (figs 9 and 14). wall of sporogenous cell composed of two permanent layers (layers 1 and 2) continuous with spore wall layers 1 and 2 (fig. 14). layer 1 pale yellow (3a3) to light orange (5a5), (1.7-)2.4(-3.2) µm thick. layer 2 cream (4a3) to light yellow (4a4), (1.5-)2.6(-3.9) µm thick at the spore base. both layers always tightly adherent to one another and difficult to observe. germination shield cardioid; pale yellow (3a3); 60-120 x 110-190 µm, of a more or less incised border; glomus eburneum and scutellospora fulgida 63 ornamented with widely usually unequally dispersed warts, 1.5-2.5 x 0.5-1.5 µm; positioned on the upper surface of the inner germination wall (fig. 15). one to three germ tubes or germ tube initials emerge from the germination shield. auxiliary cells borne in the soil, in clusters of 6-10; hyaline to cream (4a3); globose to irregular; 18-28 x 23-39 µm; with knobby projections; produced on straight or coiled hyphae; 2.5-8.0 µm diam; concolorous with auxiliary cells (fig. 16). mycorrhizal associations. the presence of mycorrhizae in field-collected root samples was not determined. many attempts to growth this fungus in one-species cultures failed. according to morton (2002), s. fulgida formed mycorrhizae with arbuscules and intraradical hyphae staining intensively in trypan blue. distribution and habitat. the spores of s. fulgida showed here were extracted from six trap cultures with rhizosphere soils and root fragments collected under plants colonizing maritime dunes of portugal (four samples), italy and oman (one sample each). the occurrence of arbuscular fungi in the field soil-root mixtures was not determined. the field samples from italy came from under ammophila arenaria (l.) link and were collected on 11 october 2002. the exact sites and dates of collections of the soil and root samples, as well as the host plants sampled in portugal and oman are unknown. the trap cultures with these samples were established on 8 march 2004 and 30 june 2003, respectively, i. e., some days after they arrived to the laboratory of the author of this paper. the arbuscular fungi accompanying s. fulgida in the cultures representing italy were acaulospora scrobiculata trappe, intraspora schenckii (sieverd. et s. toro) oehl et sieverd., glomus aurantium błaszk., v. blanke, c. renker et f. buscot, g. constrictum trappe, g. versiforme (p. karsten) s.m. berch, an undescribed glomus 169, and s. persica (koske et c. walker) c. walker et f.e. sanders. the only arbuscular fungus co-occurring with s. fulgida in the portugal culture was an undescribed glomus 172, and the culture with the rhizosphere soil and root mixture of oman still contained spores of g. fasciculatum (thaxt.) gerd. et trappe emend. c. walker et koske and s. persica. the type of s. fulgida comes from field-collected spores isolated from under a. breviligulata fern. colonizing maritime dunes of the seashore state park in virginia, u.s.a. (koske and walker 1986). this fungus has also been found in other soil samples taken from under a. breviligulata, solidago sempervirens l., and uniola paniculata l. growing in dunes extending from new jersey to virginia (koske 1987). sylvia and will (1988) found spores of s. fulgida associated with u. paniculata and panicum sp. growing in soils of a beach replenishment site in florida. additionally, s. fulgida has been reported to occur under triticum aestivum l. cultivated in argentina (schalamuk et al. 2006) and in soils of china (gai et al. 2006). collections examined. poland: szczecin, all from trap cultures with p. lanceolata as the host plant and rhizosphere soils and root fragments of a. arenaria sampled (1) near calambrone (italy; 43o35’n, 10o18’n) on 11 october 2002 and harvested on 21 june 2006, błaszkowski j. 2697-2701 (dpp); (2) in oman, harvested on 15 march 2003, błaszkowski j. 2702-2708 (dpp); and (3) in portugal, harvested on 22 july 2006, błaszkowski j. 2709-2716 (dpp). notes. the distinctive characters of s. fulgida are its light-coloured spores of a smooth surface and the sinthesization of only one inner germination wall (figs 9-13). the last property keys this fungus into a monophyletic group still comprising s. castanea c. walker, s. coralloidea (trappe, gerd. et h.h. ho) c. walker et f.e. 64 j. błaszkowski and b. czerniawska sanders, s. gregaria (n.c. schenck et nicol.) c. walker et f.e. sanders, s. persica (koske et c. walker) c. walker et f.e. sanders, and s. verrucosa (koske et c. walker) c. walker et f.e. sanders. four spore characters readily separate the species listed above. first, spores of s. fulgida are much lighter-coloured than those of the other species compared here (cream to light orange in s. fulgida (figs 9 and 10) vs. from pale straw to orange brown in s. verrucosa to red brown to dark brown in s. gregaria; morton 1995, 2002). second, in contrast to the smooth spores of s. castanea (walker, gianinazzi-pearson and marion-espinasse 1993) and s. fulgida (figs 9-14), the spore surface of the other species is ornamented with warts (morton 1995, 2002). however, s. fulgida and s. castanea markedly differ in colour and size of spores. the darkest spores of the former fungus are of a yellow shade, and mature spores of the latter species are brown (walker, gianinazzi-pearson and marion-espinasse 1993). third, although the lower size range of globose spores of s. fulgida and s. castanea overlaps, the largest spores of s. fulgida (280 µm diam) are much smaller than the greatest spores of s. castanea (up to 372 µm diam; walker, gianinazzi-pearson and marion-espinasse 1993). spores of the other species discussed here may also attain a much higher size than those of s. fulgida (384 µm diam in s. persica to 480 µm diam in s. gregaria; morton 1995). fourth, similarly as in s. castanea, the warts ornamenting the germination shield of s. fulgida spores are much lower and less densely dispersed on its surface 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clarifications and new taxa in the glomeromycota. mycol. res. 108: 979–982. walker c., trappe j. m. 1993. names and epithets in the glomales and endogonales. mycol. res. 97: 339–344. walker c., vestberg m. 1994. a simple and inexpensive method for producing and maintaining closed pot cultures of arbuscular mycorrhizal fungi. agr. sci. finland 3: 233–240. glomus eburneum i scutellospora fulgida, nowe dla europy gatunki mikoryzowych grzybów arbuskularnych (glomeromycota) s t r e s z c z e n i e opisano i zilustrowano cechy morfologiczne zarodników i mikoryz glomus eburneum oraz zarodników scutellospora fulgida, mikoryzowych grzybów arbuskularnych z gromady glomeromycota. ponadto przedstawiono poznane rozmieszczenie tych gatunków zarówno w polsce, jak i innych regionach świata. oba te gatunki nie były wcześniej podawane z europy. figs 1-8. glomus eburneum. 1. intact spores. 2. spore wall layers 1 (swl1) and 2 (swl2) and subtending hypha. 3 and 4. spore wall layers 1 (swl1) and 2 (swl2). 5. spore wall layers 1 (swl1) and 2 (swl2), subtending hyphal wall layers 1 (shwl1) and 2 (shwl2), and curved septum. 6. subtending hypha occluded by curved septum. 7 and 8. arbuscules, trunk, intraradical hyphae, and coil of mycorrhizae stained in 0.1% trypan blue. fig. 1, spores in lactic acid. figs 2, 3, 5-8, spores crushed in pvlg. fig. 4, spore crushed in pvlg+melzer’s reagent. fig. 1, bright field microscopy. figs 2-8, differential interference contrast. figs 9-16. scutellospora fulgida. 9. intact spores with bulbous sporogenous cells. 10-13. spore wall layers 1 (swl1) and 2 (swl2) and inner germination wall layers 1 (gwl1) and 2 (gwl2). 14. spore wall layers 1 (swl1) and 2 (swl2) and sporogenous cell wall layers 1 (scwl1) and 2 (scwl2). 15. germination shield with germ tube and two germ tube initials. fig. 9, spores in lactic acid. figs 10 and 16, spores crushed in pvlg. figs 11-15, spores crushed in pvlg+melzer’s reagent. fig. 9, bright field microscopy. figs 10-16, differential interference contrast. 2014-01-01t11:47:14+0100 polish botanical society studies on ectomycorrhizal basidiomycete in pine forest on the lithuania–poland transboundary region danutė stankevičienė and jonas kasparavičius laboratory of mycology, institute of botany žaliųjų ežerų str. 49, lt-08406 vilnius, mikods@botanika.lt s t a n k e v i č i e n ė d . , k a s p r a v i č i u s j . : studies on ectomycorrhizal basidiomycete in pine forest on the lithuania–poland transboundary region. acta mycol. 42 (1): 59-68, 2007. the diversity of ectomycorrhizal fungi and sporocarps abundance were investigated in 20032005 at nine permanent study plots in a 50-year-old pine forest. diversity of ectomycorrhizal fungi consist of 53 taxa and the majority of them belonged to the genera cortinarius, russula, amanita and tricholoma. the most frequent species, whose fruit bodies were found in each study plot, were c. cibarius, l. necator l. rufus, p. involutus, r. aeruginea, t. saponaceum and the most abundant species which made the main part of total sporocarp yield were c. cibarius and p. involutus. the lowest species richness of ectomycorrhizal fungi was in study plots with the densest cover of grasses. maximum of species over the fruiting period was characteristic for october and for september. it was noticed that some species virtually never occurred together at the same plot (e.g. c. cibarius and h. aurantiaca), meanwhile others occurred together quite frequently (e.g. h. aurantiaca and x. badius). key words: ectomycorrhizal fungi, species richness, sporocarps abundance, pine forest introduction wild mushrooms are becoming more important as a non-timber forest product and there is a need for more site-specific data on the fungi ecology and factors that influence species diversity and production of sporocarp. macrofungi especially ectomycorrhizal ones are organisms vitally important to the forest ecosystem. ectomycorrhizae (em) plays a key role in nutrient cycling and energy flow of temperate and boreal forests (s m i t h , r e a d 1997). the best known mycobionts of em belong to the basidiomycota. host specificity plays an important role for the distribution of ectomycorrhizal fungi. under natural condition wide range of ectomycorrhizal fungi develop ectomycorrhizal symbiosis with pinus sylvestris. p. sylvestris is one of the main components of coniferous forests in lithuania. conifer make 58.8 % of lithuanian woodland territory, 36.4 % of the territory is occupied by p. sylvestris and 22.4 % by picea abies (n a v a s a i t i s et al. 2003). variations in fungal species richness, distri acta mycologica vol. 42 (1): 59-68 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 60 d. stankevičienė and j. kasparavičius bution, and sporocarp abundance among different forest sites have been observed and may be attributed to microclimatic and macroclimatic factors, soil properties, vegetation parameters etc. forest age has been observed to be an important factor determining the composition of ectomycorrhizal fungi (d i g h t o n , m a s o n 1985; m o l i n a et al. 1992; o h e n o j a 1993; d a h l b e r g et al.1997). we investigated assemblage structure of ectomycorrhizal fungi associated with 50–years–old p. sylvestris. the objectives of present study were: 1) to perform inventory of ectomycorrhizal fungus diversity, 2) to examine sporocarp abundance of ectomycorrhizal fungi aiming to determine dominant species in the investigation territory, 3) to obtain a quantitative estimate of the relative contributions of dominant ectomycorrhizal species to assemblage structure in 50-years-old p. sylvestris forest situated in lithuanian– poland transboundary region. materials and methods study site. the study was carried out in permanent study site, situated in lazdijai district, southern lithuania (125 – 135 m a.s.l.) (fig. 1). the mean air temperature was 6.4o c and mean annual precipitation – 550 mm. this territory is located in lithuanian – poland transboundary region were access for people is prohibited. this factor is important for obtaining objective investigation data because most of the ectomycorrhizal fungi are edible and intensively collected. nine study plots (1 – 9) were set in the 50 year-old pine forest of the cladonio-pinetum sylvestris juraszek 1927 association. area of each study plot was 900 m2 (30x30 m). dominant tree species was pinus sylvestris l. in some locations betula pendula roth., quercus robur l. were intermixed. the shrub layer was predominantly by juniperus communis l., with fig. 1. location of study area●. ectomycorrhizal basidiomycete 61 occasional q. robur, frangula alnus mill., sorbus aucuparia l. about 80 % of study area was occupied by mosses and lichens (tab. 1). dominant species of mosses were pleurozium schreberi (brid.) mitt., dicranum polysetum sw., lichens – cladonia rangiferina (l.) weber ex f. h. wigg., cladonia arbuscula (wallr.) flot. qualitative and quantitative analyses of ectomycorrhizal fungi. species composition of fungi was inventoried at each selected forest plot every second or third week during the vegetation period in 2003 – 2005. investigation started at the beginning of june and lasted until the first snowfall. identification of specimens was carried out according to m o s e r (1983), s k i r g i e ł ł o (1991, 1998), h a n s e n and k n u d s e n (1992), u r b o n a s (1997, 2001, 2005) using a microscope “jenaval carlzeiss jena”. voucher specimens of infrequent species found within this study are deposited in the fungal collections of the herbarium of the institute of botany (bilas). number of fruit bodies collected in each investigation plot was counted. sporocarps were weighted and biomass of fresh sporocarps (kg/ha) was calculated. soil analyses. soil samples for chemical analyses were taken with soil corer of 4.5 cm diameter and 6 cm depth in august of each investigation year. the representative sample for each research plot was prepared of 18-20 randomly taken sub-samples from each plot. these soil samples were dried and sieved before the following analyses. the concentration of nitrogen and phosphorus was determined photometrically using the photometer “spekol11”, of potassium – applying flame photometer “flapho41”, and the content of humus was ascertained colorimetrically (m i n e e v 1989). soil phkcl was measured potentiometrically with a glass electrode in a 1.0 m kcl suspension. meteorological data were obtained from the state meteorological service. data analysis was carried out using the software pc-ord4 (m c c u n e , m e f f o r d 1999). results and discussion chemical characteristics of soil. chemical composition of soil directly influences functioning of fungi in the ecosystem. therefore the concentration of main nutrient elements – n, p, k, humus and soil ph was determined. analysing obtained data some differences were observed between study plots. the highest concentration of n, k and humus was determined in the 9. investigation plot (tab. 2). these concentrations were several times higher comparing to the other study plots. 9. plot is situated in the lowermost position of study area and perhaps this resulted in such a soil composition. the 8. plot distinguished by the lowest concentration of p. higher ta b l e 1 vegetation cover (%) of study plots (evaluation according to braun-blanquet method) vegetation groups plots 1 2 3 4 5 6 7 8 9 trees 50 60 60 60 50 50 60 70 60 shrubs 60 50 10 10 10 10 15 15 20 herbaceous plants 50 30 10 10 10 10 20 20 40 mosses, lichens 80 70 80 70 70 70 70 80 80 62 d. stankevičienė and j. kasparavičius concentration of this element was determined in the 2. and 7. plots. this is especially important for ectomycorrhizal fungi because concentration of nutrients alters ectomycorrhizal formation and community structure (a v i s et al. 2003; s t a n k e v i č i e n ė 2003; ta r v a i n e n et al. 2003; e d w a r d s et al. 2004; h a r r i n g t o n , m i t c h e l l 2005). other values were more or les similar comparing them between study plots. diversity of ectomycorrhizal fungi and sporocarp abundance. the diversity of ectomycorrhizal fungi recorded in 2003-2005 at nine permanent study plots consisted of 53 taxa (tab. 3). most species belonged to the genus cortinarius – 12 species (23%) and russula –10 (18%). five species were found of amanita and tricholoma genera, four lactarius and three hebeloma of. other genera were represented by 1-2 species. cantharellus cibarius, lactarius necator, l. rufus, paxillus involutus, russula aeruginea and tricholoma saponaceum were found in each study plot. ta y l o r (2002) investigating diversity of ectomycorrhizal fungi in central sweden in a 50–year-old pinus sylvestris stand found very similar species richness 56 species. species of ectomycorrhizal basidiomycetes from the investigated pine forest could be distributed into several groups according to the abundance of formed fruit bodies. 22 species formed only small amount of fruit bodies – 10 sporocarps/investigation period. rarest species, of which only 1 – 2 fruit bodies/investigation time were found, were b. pinophilus, c. causticus, c. bovinus, c. delibutus, c. evernius, h. pusila, l. bicolor, l. vietus, r. claroflava, r. decolorans and t. felleus. seven species were the most abundant and formed more than 100 fruit bodies in study plots/ investigation time. it was c. cibarius, p. involutus, rozites caperata, l. rufus, hygrophoropsis aurantiaca, cortinarius mucosus and r. aeruginea. those were dominant species in 50-year-old pine forest. intermediate group was composed of 24 species which formed 11-100 fruit bodies in study area per investigation period. it is known that each forest type has its own dominant mushroom species and that dominants make main biomass of sporocarps and usually determine harvest in different forest types (s k r y a b i n a , s e n n i k o v a 1981). c. cibarius was the most abundant species, its sporocarps made about a half of the total numbers of all collected sporocarps of ectomycorrhizal fungi. p. involutus harvested quite abundantly also. the number of fruit bodies formed by this species made about a quarter and their biomass – about a half of the total amount of fruit bodies. studies on fungal fruit bodies in mixed ta b l e 2 chemical composition of soil (dw) from the study plots plots n (%) p (%) p2o5 (mg/kg) k (mg/kg) humus (%) ph (kcl) 1 0,073 0,027 116,8 38,3 3,75 3,82 2 0,039 0,035 150,2 36,1 3,79 3,76 3 0,035 0,027 85,8 27,7 2,94 3,78 4 0,029 0,034 118,4 21,0 2,71 3,73 5 0,079 0,026 100,8 31,2 2,89 3,66 6 0,075 0,029 107,1 26,1 2,91 3,69 7 0,033 0,037 132,7 18,9 2,05 3,82 8 0,058 0,014 49,4 29,3 2,97 3,43 9 0,197 0,028 104,6 81,0 8,02 3,47 ectomycorrhizal basidiomycete 63 ta b l e 3 diversity of ectomycorrhizal fungi species and sporocarp abundance in study forest species species code sum 1 mean 2 maximum 3 s 4 amanita citrina (schaeff.) pers. amcitr 51 5.7 23 4 a. fulva (schaeff.) fr. amfulv 26 2.9 25 2 a. muscaria (l.) lam. ammusc 53 5.9 35 7 a. porphyria fr. amporf 28 3.1 12 5 a. rubescens pers. amrube 42 4.7 15 7 boletus edulis bull. boledu 18 2 10 5 b. pinophilus pilát et dermek bolpin 2 0.2 1 2 cantharellus cibarius fr. cantc 5526 614 1095 9 cortinarius alboviolaceus (pers.) fr. cortal 4 0.4 2 2 c. armillatus (alb. et schwein.) fr. cortam 11 1.2 7 2 c. causticus fr. cortca 1 0.1 1 1 c. bolaris (pers.)fr. cortbo 3 0.3 3 1 c. bovinus fr. cortbv 1 0.1 1 1 c. cinnamomeus (l.) fr. cortci 5 0.6 3 2 c. delibutus fr. cortde 2 0.2 2 1 c. evernius (fr.) fr. cortev 2 0.2 2 1 c. mucosus (bull.) cooke cortmu 146 16 45 8 c. salor fr. cortso 3 0.3 3 1 c. semisanguineus (fr.) gillet cortse 5 0.6 4 2 c. traganus (fr.) fr. corttr 44 4.9 18 6 hebeloma crustuliniforme (bull.) quél. hebecr 44 4.9 44 1 h. longicaudum (pers.) p. kumm. hebelo 3 0.3 3 1 h. pussilum j. e. lange hebepu 1 0.1 1 1 hygrophoropsis aurantiaca (wulfen) maire hygaur 173 19 65 7 laccaria bicolor (maire) p. d. orton lacbic 2 0.2 2 1 lactarius necator (bull.) pers. lactne 96 10.7 30 9 l. rufus (scop.) fr. lactru 392 43.6 104 9 l. torminosus (schaeff.) gray lactto 28 3.1 20 2 l. vietus (fr.) lactvi 1 0.1 1 1 leccinum scabrum (bull.) gray leccsc 19 2.1 7 8 paxillus involutus (batsch.) fr. paxinv 2475 275 552 9 rozites caperata (pers.) p. karst. rozcap 405 45 117 7 russula adusta (pers.) fr. rusadu 24 2.7 20 3 r. aeruginea fr. rusaer 99 11 29 9 r. claroflava grove ruscla 1 0.1 1 1 r. decolorans (fr.) fr. rusdec 2 0.2 1 2 r. emetica (schaeff.) pers. ruseme 79 8.8 24 7 r. nigricans (bull.) fr. rusnig 3 0.3 2 2 r. rhodopoda zvára rusrod 14 1.6 9 5 r. sanguinea (bull.) fr. russan 3 0.3 3 1 r. vesca fr. rusves 32 3.6 13 8 r. xerampelina (schäff. ex secr.) fr. rusxer 4 0.4 4 1 sarcodon imbricatus (l.) p. karst. sarimb 92 10.2 42 5 suillus bovines (pers.) kuntze suilbo 27 3 14 6 s. variegatus (sw.) kuntze suilva 24 2.7 11 5 tylopilus felleus (bull.) p. karst. tylofe 1 0.1 1 1 tricholoma equestre (l.) p. kumm. triequ 90 10 27 5 t. pesundatum (fr.) quél. tripes 4 0.4 3 2 t. portentosum (fr.) quél tripo 40 4.4 14 7 t. saponaceum (fr.) p. kumm. trisap 78 8.7 32 9 t. sejunctum (sowerby) quél. trisej 9 1 4 3 xerocomus badius (fr.) kühner xerbad 85 9.4 40 8 x. subtomentosus (fr.) fr. xersub 34 3.8 19 7 explanations: 1 – sum of sporocarps collected in all study plots/whole investigation period (2003–2005); 2 – mean of sporocarp number yielded in one plot; 3 – maximum of sporocarps yielded in one plot; 4 – number of plots in which the species yielded. 64 d. stankevičienė and j. kasparavičius forest in switzerland showed that abundance of ectomycorrhizal sporocarps varied over the years of the investigation and ranged from 58 to 5559 per vegetation season (s t r a a t s m a e t a l . 2001). present studies showed that sporocarp abundance of ectomycorrhizal fungi in pine forest in lithuania over the study period varied from 2487 to 5025 sporocarps per year. a total of 10808 fruit bodies of ectomycorrhizal basidiomycetes during the three year study period were collected. it made 894 kg (fresh weight)/8100m2 or 1104 kg/ha per investigation time or 368 kg/ha per one vegetation season. according to the sporocarp abundance study plots were distributed into three groups. about 1,500 fruit bodies per investigation period were collected in the richest plots (4, 5, 8). plots 1 – 3, 6 and 9 took intermediate position and yielded 1000 – 1270 fruit bodies. the least amount, a little over 300 sporocarps was found in the plot 7. it was 3-5 times lower than in the other study plots. as it was mentioned above, c. cibarius was the dominant species in the investigated pine forest. however, amount of fruit bodies of chanterelle in the plot 7 was very low (only 9 fruit bodies/ investigation period) and this determined the minimum total yield of sporocarps in this plot. meanwhile h. aurantiaca and xerocomus badius fruited in this plot most abundantly. it is interesting that the reliable positive correlation (r = 0.7) between the abundance of sporocarps of the last-mentioned species and reliable negative correlation between c. cibarius and h. aurantiaca (r = -0.70) also c. cibarius and x. badius (r = -0.76) was determined. analysing distribution of species (these which yielded more than 5 fruit bodies per investigation period) and their frequency in study plots several groups were selected (fig. 2). it means that is likely to find species which belong to the same group (e.g. c. cibarius, r. vesca, also a.porphyria, s. bovinus etc.) fig. 2. groups of ectomycorrhizal fungi species (yielded more than 5 fruit bodies per investigation time) which frequently occurred together at the same plot. ectomycorrhizal basidiomycete 65 in the same area with the similar relative abundance and on the contrary the species from different groups prefer different growing conditions. these observations were made after only three year field study. aiming to confirm this relationship the investigation time possibly should be prolonged and at present it can be stated more as a tendency than the strong correlation. a g e r e r et al. (2002) while investigating correlations between ectomycorrhyzae formed by different ectomycorrhizal fungi species noted that some species show no common occurrence within shot distance, however, other indicate rather high values of common occurrence. the reasons of this phenomenon are not quite clear. the chemical composition of soil is an important factor, because varying demands of different species for soil conditions are expressed. it was determined that changes of different soil ions concentrations (n, p, s, k, na, ph etc.) influence aboveand belowground community structure of ectomycorrhizal fungi (ty l e r 1985; a g e r e r 1990; e r l a n d , s ö d e r s t r ö m 1990; f r a n s s o n et al. 2000; l i l l e s k o v et al. 2002; a g e r e r , g o t l e i n 2003, ta r v a i n e n et al. 2003; s t a n k e v i č i e n ė , u r b o n a s 2006). in the present studies the plot with the minimum amount of fruit bodies was distinguished by the lowest concentration of k, humus and the highest value of ph (tab. 2). the lowest species richness (15 species) was determined in the plot 1 which characterized by the highest coverage of herbaceous plants (grasses) (50 %) and shrubs (60 %) (tab. 1). on the other hand, the plots with the maximum species (4. plot – 32 species; 9. – 30; 3. – 29) were characterized by the lower coverage of shrubs (10 %) and grasses (10 %, except plot 9, where coverage of grasses was 40 % and shrubs – 20 %). fungal species composition seems to be strongly determined by soil chemical properties, vegetation type, structure and age of the forest stand especially in the case of ectomycorrhizal species. phenology. fruit bodies were monitored every second or third week between june and october. the start of fruiting varied strongly between the species. the longest period of fruiting was characteristic to c. cibarius and p. involutus. fruit bodies of these species started to growth in june and fruited till the end of the vegetation season. long fruiting period was characteristic also for l. scabrum, b. edulis, also for some species from genus amanita, lactarius, russula, xerocomus. fruit bodies of mentioned fungi started to growth in july or august. species from genus cortinarius, tricholoma, hebeloma also sarcodon imbricatus were found from september and their fruiting period was relatively short. maximum species richness was characteristic for september in 2004 and for october in 2003, 2005. the dynamics curve of richness and abundance per vegetation season (june-october) was sinusoid (fig. 3 a, b). it means that minimum value of species or sporocarps in one period (eg. a month) was compensated by the maximum in the next period and, on the contrary, the maximum was replaced by the minimum values in the following fruiting period. the dynamic of the species richness and fruit bodies abundance in 2003 and 2004 demonstrated similarities of these parameters in different fruiting periods. however, comparing appointed values between the three year period, it was noted that the curve of 2005 quite differed from 2003, 2004. the reason of these differences probably was meteorological conditions, because the curves of the dynamic of species richness, sporocarp abundance and precipitation demonstrated similar patterns in the period of 2003 – 2005 (fig. 3 a, b, c). it is known that the highest ectomycorrhizal basidiomycete 67 references a g e r e r r. 1990. impact of acid rain and liming on fruit body production of ectomycorrhizal fungi. agric. ecosyst. 28: 3–8. a g e r e r r , g o t l e i n a . 2003. correlations between projection area of ectomycorrhizae and h2o extractable nutrients in organic soil layers. mycological progress 2 (1): 45–52. a g e r e r r . , g r o t e r . , r a i d l s . 2002. the new method micromapping, a means to study speciesspecific associations and exclusions of ectomycorrhizae. mycological progress 1 (2): 155–166. a v i s p. g . , a c l a u g h l i n d . j . , d e n t i n g e r b . c . , r e i c h p. b . 2003. long-term increase in nitrogen supply alters aboveand below-ground ectomycorrhizal communities and increases the dominance of russula spp. in a temperate oak savanna. new phytologist 160: 239–253. d a h l b e r g a . , j o n s s o n l . , n y l u n d j . 1997. species diversity and distribution of biomass above and below ground among ectomycorrhizal fungi in an old-growth norway spruce forest in south sweden. can. j. bot. 75: 1323–1335. d i g h t o n j . , m a s o n p. 1985. mycorrhizal dynamics during forest tree development. 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(in:) m . j . a l l e n (ed.). mycorrhizal functioning, an integrative plant-fungal process. chapman and hall, new york: 357–423. m o s e r m . 1983. die röhrlinge und blätterpilze (polyporales, boletales, agaricales, russulales). fischer verlag, stuttgart. n a v a s a i t i s m., o z o l i n č i u s r., s m a l i u k a s d., b a l e v i č i e n ė j . 2003. lietuvos dendroflora (lithuanian dendroflora). kaunas. o h e n o j a e . 1993. effect of weather conditions on the larger fungi at different forest site in northern finland in 1976–1988. acta universitatis ouluensis a 234: 1–69. s e n n i k o v a l. s. 1984. urozhaynost’ s’edobnykh gribov v kirovskoy oblasti. mikologiya i fitopatologiya 18(6), 455–459. s k i r g i e ł ł o a . 1991. flora polska. grzyby (mycota). 20. gołąbek (russula). pwn, warszawa. s k i r g i e ł ł o a . 1998. flora ploska. grzyby (mycota). 25. mleczaj (lactarius). w. szafer institute of botany, polish academy of sciences, kraków. s k r y a b i n a a. a., s e n n i k o v a l. s. 1981. ekologicheskiye osobennosti plodonosheniya s’edobnykh gribov v lesnykh cenozakh severo-vostoka evropeiskoy chasti sssr. (in:) biologicheskiye problemy severa. syktyvkar: 456–459. 68 d. stankevičienė and j. kasparavičius s m i t h s. e., r e a d d. j 1997. mycorrhizal symbiosis. academic, london. s t a n k e v i č i e n ė d . 2003. ectomycorrhizae in a deciduous forest near a factory of chemical fertilizes. baltic forestry 9 (1): 43–49. s t a n k e v i č i e n ė d., u r b o n a s v. 2006. diversity of agaricoid fungi and ectomycorrhizae in deciduous forest along pollution gradient. mycologiya i fitopatologiya 40 (2): 108–116. s t r a a t s m a g., a y e r f., e g l i s . 2001. species richness, abundance, and phenology of fungal fruit bodies over 21 years in a swiss forest plot. mycol. res. 105 (5): 515–523. ta r v a i n e n o., m a r k k o l a a. m., s t r o m m e r r . 2003. diversity of macrofungi and plant in scots pine forests along an urban pollution gradient. basic appl. ecol. 4: 517–556. u r b o n a s v. 1997. baltikiečiai (tricholomatales). lietuvos grybai (mycota lithuaniae) 8 (2). vilnius. u r b o n a s v. 2001. musmiriečiai (amanitales), ūmėdėčiai (russulales). lietuvos grybai (mycota lithuaniae) 8 (4). vilnius. u r b o n a s v. 2005. nuosėdiečiai (cortinariales). lietuvos grybai (mycota lithuaniae) 8 (5). vilnius. ta y l o r a. f. s. 2002. fungal diversity in ectomycorrhizal communities: sampling effort and species detection. plant and soil 244: 19–28. ty l e r g. 1985. macrofungal flora of swedish beech forest related to soil organic matter and acidity characteristics. forest ecology and managament 10: 13–29. 2014-01-01t11:45:10+0100 polish botanical society three rare lignicolous fungi from sicily (s italy) giuseppe venturella1, annarosa bernicchia2 and alessandro saitta1 1department of botany, university of palermo via archirafi 38, i 90123 palermo, gvent@unipa.it 2dipartimento di scienze e tecnologie agroambientali, università di bologna viale fanin 44, i 40127 bologna, abernicc@agrsci.unibo.it ve n t u r e l l a g., b e r n i c c h i a a., s a i t t a a.: three rare lignicolous fungi from sicily (s italy). acta mycol. 41 (1): 95 98, 2006. sarcodontia crocea (schwein.) kotl., oligoporus mappa (overh. & lowe) gilbn. & ryvarden and inonotus rickii (pat.) d.a. reid. are reported for the first time from sicily (southern italy). ecological and distributive features on the three relevant species are also here provided. key words: lignicolous fungi, ecology, distribution, sicily, mediterranean area introduction the increase of mycological investigation in sicily permitted to select a huge number of fungi, to publish lists and maps of distribution, to collect many ecological data and to evaluate the rareness of some taxa. a specific field of investigation was devoted to lignicolous fungi which are widely distributed in sicily on different plants of agronomic, forestry and ornamental interest. a list of lignicolous species, host plants and substrata, referred to 209 taxa, was recently published by s a i t t a et al. (2004). besides distributive and ecological data on eleven aphyllophorales were reported by ve n t u r e l l a et al. (in press). in this paper the results of field investigation on three lignicolous fungi, considered as rare and/or infrequent in italy, are pointed out. materials and methods in the framework of a project for the assessment of fungal biodiversity in sicily, observations on lignicolous fungi were carried out in different forest ecosystems, parks and botanical gardens. the basidiomata were collected with a knife, preserved in a paper bag, and identified in laboratory, after rehydratation in koh, using a leica microscopy. the monographies of b e r n i c c h i a (2005); e r i k s s o n et al. (1981) and h j o r t s t a m et acta mycologica vol. 41 (1): 95-98 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 96 g. venturella et al. al. (1987) were consulted for identification and nomenclatural notes. the herbarium samples were prepared in a dryer and kept in the herbarium mediterraneum (pal). the distributive data were referred to the grid map 1:50,000 of the official map of the italian state (i.g.m.i.), following the methodology proposed by padovan (1994) for the checklist of macromycetes in italy. in particular the sheets number 595 (palermo), 609 (termini imerese) and 620 (lercara friddi) were divided into 64 subunits. consequently, a number composed by six figures were attributed to each recorded taxon. results sarcodontia crocea (schwein.) kotl. specimen examined. sicily (s italy). piano torre (province of palermo), 950 m, quercus ilex l. wood, on stumps of q. ilex, 01 october 2005, 609122, leg. a. saitta (fig. 1a, b). comments. this species was gathered on stumps in a q. ilex wood located within the madonie park, a protected area in the province of palermo (n sicily). in the locality of collection the forest management, carried out by the forestry administration, lead to accumulation of a huge amount of logs, branches, stumps and rotten woods which are easily overrun with rot decay fungi. s. crocea, previously listed in italy from emilia romagna, friuli venezia giulia and lombardia and collected on old apple-trees only, is reported for the first time from sicily. this new finding extend southwards the area of distribution of s. crocea and confirm the presence of such species on different broad-leaved host plants. it is also interesting to be note that s. crocea is known from finland, sweden, denmark, poland, germany, the netherlands and u.k., but it is considered threatened in all these countries. in estonia it is listed as extinct (l æ s s ø e 2004). the rarity of s. crocea is probably caused by the progressive abandonment of tilled fields and the decrease in the number of old apple-trees. oligoporus mappa (overh. & lowe) gilbn. & ryvarden specimen examined. sicily (s italy). monte rose (province of palermo), 1000 m, mixed reafforestation of pinus halepensis miller and cupressus sempervirens l., on trunks of c. sempervirens, 26 september 2004, 620313, leg. c. vasile (fig. 2a, b). comments. o. mappa grows as saprotroph on conifer woods, rarely on broad-leaved plants, and sometimes on manufactured wood. the only report of such species for italy arise from bernicchia (1990) and is referred to a collection on juniperus phoenicea l. in foresta montes (supramonte di orgosolo) in the province of nuoro (sardegna). o. mappa was recently recorded in sicily, in a mixed reafforestation of p. halepensis and c. sempervirens, on fallen trunks of c. sempervirens. it is also interesting to note that the new locality here reported is located at the same altitude, i.e. 1000 m, of the previous locality pointed out by b e r n i c c h i a (1990). o. mappa is a very rare species in europe but widely distributed. three rare lignicolous fungi from sicily 97 inonotus rickii (pat.) d.a. reid specimen examined. sicily (s italy). botanical garden (town of palermo), 3 m, on stumps of acer negundo l., 23 october 2003, 595433, leg. c. comparetto (fig. 3a, b). comments. the first finding for europe was reported by j a q u e n o u d (1985) as anamorph state on parkinsonia aculeata l. in villa giulia, a public garden of the town of palermo (sicily). a second collection, on schinus molle l., from sicily was published by i n t i n i (1988) recently, i n t i n i (2002) gathered i. rickii on acer negundo l. in tree plantations along the streets of sevilla (spain). i. rickii was also found by a n n e s i et al. (2003) on a. negundo and celtis australis l. in rome and by saitta et al. (2005) on a. negundo in villa giulia (palermo). typical brown, powdery masses of chlamydospores of i. rickii were observed near wounds or pruning cuts on quercus cerris l. in the botanical garden of palermo and on sambucus nigra l. and aberia caffra hook. f. & harv. in catania (a n n e s i et al. 2005). i. rickii is also reported from montenegro, greece and france (b e r n i c c h i a 2005) and morocco (b e r n i c c h i a , in litteris). conclusions the distribution of lignicolous fungi in italy is still not well known and the availability of maps mainly depends on the presence of mycologists in the different regions of the country. the application of the term “rare” in mycology is not suitable for fungi recorded only one time or in absence of long term researches but, in some cases, the results of field investigation, the availability of ecological and literature data allowed to evaluate the rareness of species belonging to taxonomic group not well investigated. this is the very case of sarcodontia crocea, oligoporus mappa and inonotus rickii which are recognized as rare or threatened at european level. the progressive abandonment of tilled fields, the decrease in the number of old cultivars and the disappearance of specific host plants are the main factors responsible of decline in lignocolous species together with unsuitable forest managements particularly in protected areas. the finding of s. crocea, o. mappa and i. rickii contributed to widen the area of distribution in europe and to provide additional ecological data. acknowledgements. the authors wish to thank dr david minter for kindly revised this manu script. references a n n e s i t., c o p p o l a r., m o t t a e. 2003. decay and canker caused by inonotus rickii spreading on more urban tree species. for. path. 33: 405 412. a n n e s i t., c o p p o l a r., d ’ a m i c o l., m o t t a e. 2005. first report of aberia caffra and quercus cerris as hosts of inonotus rickii. plant. dis. 89: 107. b e r n i c c h i a a. 1990. polyporaceae s.l. in italia. istituto di patologia vegetale, università degli studi di bologna. b e r n i c c h i a a. 2005. polyporaceae s.l. fungi europaei, 10. ed. candusso, savona (italia). e r i k s s o n j., h j o r t s t a m k., r y v a r d e n l. 1981. the corticiaceae of north europe. vol. 6. phlebia sarcodontia. fungiflora, oslo, norway. 98 g. venturella et al. h j o r t s t a m k., l a r s s o n k.h., r y v a r d e n l. 1987. the corticiaceae of north europe. introduction and keys. vol. 1. fungiflora, oslo, norway. i n t i n i m.g. 1988. contributo alla conoscenza dei funghi lignicoli italiani: inonotus rickii (pat.) reid. micol. ital. 17 (1): 49 53. i n t i n i m.g. 2002. first report of inonotus rickii causing canker rot on boxelder in europe. plant dis. 86: 922. j a q u e n o u d m.1985. inonotus rickii, un polypore nouveau pour la flore européenne. mycologia hel vetica i (6): 371 391. l æ s s ø e t. 2004. æblepig (sarcodontia crocea) nu fundet i danmark. i: vesterholt, j. usædvanlige danske svampefund. svampe 49: 40 42. p a d o v a n f.1994. mappatura dei macromiceti in italia (problemi cartografici). rivista di micologia 37 (1): 59 69. r y v a r d e n l., g i l b e r t s o n r. l. 1993. european polypores. fungiflora, oslo (norway). s a i t t a a., b e r n i c c h i a a., ve n t u r e l l a g. 2004. contributo alla conoscenza dei funghi lignicoli della sicilia. inform. bot. ital. 36 (1): 192 202. s a i t t a a., p e c o r e l l a e., c o m p a r e t t o c. 2005. 9 inonotus rickii (pat.) d. a. reid. p. 191. (in:) ve n t u r e l l a g. , r a i m o n d o f. m. (eds). i funghi cariogeni delle alberature di parchi, giardini e strade 1 18. quad. bot. ambientale appl. 15(2004): 181 201. ve n t u r e l l a g., b e r n i c c h i a a., s a i t t a a. contribution to the knowledge of diversity and distribu tion of lignicolous fungi from sicily (southern italy). bocconea (in press). trzy rzadkie gatunki grzybów nadrzewnych z sycylii s t r e s z c z e n i e autorzy podają trzy nowe dla sycylii gatunki grzybów: sarcodontia crocea, oligoporus mappa i inonotus rickii. dla każdego gatunku podana jest chrakterystyka ekologiczna, barwna fotografia i mapa rozmieszczenia na terenie włoch. 2014-01-01t11:43:43+0100 polish botanical society genetic diversity of heterobasidion spp. in scots pine, norway spruce and european silver fir stands piotr łakomy1, zbigniew broda2 and antoni werner3 1department of forest pathology, the august cieszkowski university of agriculture wojska polskiego 71c, pl-60-625 poznań, plakomy@au.poznan.pl 2department of genetics and plant breeding, the august cieszkowski university of agriculture wojska polskiego 71c, pl-60-625 poznań 3institute of dendrobiology, polish academy of sciences, parkowa 5, pl-62-035 kórnik ł a k o m y p., b r o d a z., we r n e r a.: genetic diversity of heterobasidion spp. in scots pine, norway spruce and european silver fir stands. acta mycol. 42 (2):203-210, 2007. investigations of genetic diversity of heterobasidion spp. in scots pine, norway spruce and european silver fir stands indicated that almost all of identified genets occurred in those stands were small occupied only a single stump. in some cases two, three or even four genets could effectively exist in an individual stump. genetic similarity of h. annosum s.s. genets varied from 0% to 62%, h. parviporum from 0% to 38% and h. abietinum from 0% to 55%. the oldest and biggest genet was found in laying fir log and overgrew the wood for at least 14 years. this genet belonged to h. abietinum. the size of genets was related to thinning operation, spore dispersal, age of stand or competition in wood colonization. key words: heterobasidion annosum sensu stricto, h. parviporum, h. abietinum, genets, pine, spruce, fir introduction root and butt rot caused by heterobasidion spp. is the most important disease in forests. in europe there were described three species of heterobasidion – h. annosum (fr.) bref sensu stricto, heterobasidion parviporum niemelä and korhonen, heterobasidion abietinum niemelä and korhonen (n i e m e l ä , k o r h o n e n 1998). in poland the most economic losses causes h. annosum sensu stricto in scots pine plantations growing on post-agricultural lands (s i e r o t a 1987, 1995; m a ń k a 2005). distribution of heterobasidion species is related to the natural range of their main hosts. heterobasidion annosum sensu stricto, which attack mainly scots pine (pinus sylvestris l.), occurs almost in whole poland. heterobasidion parviporum infected almost only norway spruce (picea abies (l.) karst.) is noted in south and northeast part of poland and heterobasidion abietinum appeared on european silver fir (abies alba mill.) stumps or laying logs in south poland. those three species could acta mycologica vol. 42 (2): 203-210 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 204 p. łakomy et al. infect more than 200 species also deciduous tree as birch, beach or oak (we b b , a l e x a n d e r 1985; ł a k o m y et al. 2000; ł a k o m y , we r n e r 2003; ł a k o m y , c i e ś l a k 2006). heterobasidion spp. is dispersed mainly by basidiospores, which colonise freshly cut stumps or wounds (r e d f e r n and s t e n l i d 1998). in colonized stand pathogens could spread vegetatively by mycelium transfer via roots contacts between colonized and healthy tissue (s t e n l i d , r e d f e r n 1998). the affected stumps or trees became the source of spore production. appearance of new genets in stands is related to new spore infection, thinning operations and pathogen’s frequency and in particular basidiocarps on stumps or trees. almost sixty percent genets of heterobasidion sp. identified in final cutting stands had infected only one tree (p i r i et al. 1990; va s i l i a u s k a s , s t e n l i d 1998). however, bigger genets were also described, which occurs in 10 or more trees (s t e n l i d 1985; p i r i , k o r h o n e n 2001). small size of genets might be resulted by lacking of roots contact and impossibility of mycelium transfer among trees and stumps. it could also indicate that the genets are young or very old and are not able for spreading. the small size of genets could also indicate a high competition among genets in a single stump. the aim of this study was i) to enter the investigations of distribution of heterobasidion spp. genets in scots pine, norway spruce and european silver fir stands in poland and ii) to investigate the genetic similarity of genets. materials and methods three stands were chosen to this study: 53-year-old picea abies stand (suwałki forest district 54o17’n, 22o51’e), 37-year-old pinus sylvestris stand (skwierzyna forest district 52o33’n, 15o22’e) and 180-year-old abies alba stand (siemianice experimental forest of august cieszkowski agricultural university in poznań, 51o01’n, 18o05’e). norway spruce and scots pine stands were growing as a first generation on post-arable soil. in fir stand two groups of stumps were chosen for this study (7 stumps and laying log and 5 stumps). the two groups were remote each other about 250 m. in each stand, stumps were investigated for presence of decay or heterobasidion sp. basidiocarps. wood from stumps was collected. in laboratory the mycelium of heterobasidion sp. were isolated from wood samples. the genets were identified with the aid of somatic incompatibility test (s t e n l i d 1985). cultures were pairing in all possible combinations on 1.5% malt extract agar (merck, germany). after 2-3 weeks pairings were marked to presence the demarcation line indicating different genets. isolates representatives of each genet were identified to heterobasidion species with the compatibility test (k o r h o n e n 1978). genetic analysis. rapd analysis were carried out using dna, extracted as by t h o m p s o n and h e n r y (1995) from genotypes of h. annosum ss., h. parviporum and h. abietinum. dna of each genotype was extracted from 10mm2 mycelium, soaked for 15 min at 95oc in 120 μl of tps buffer. pcr reactions (12,5 μl) contained 10 ng of genomic dna, 0,94 u taq polymerase, 1.25 pmol of primer, 2mm of dntp, 1.25 μl of bsa, 25 mm mgcl2 and tris-hcl. six oligonucleotide primers (10 bases long) were used in each pcr amplification. genetic diversity 205 the cycling was performed as follows: 94oc/60 s followed by 10 cycles of amplification (94oc/5 s, 37oc/30 s, 72oc/30 s) and the next 35 cycles (94oc/5 s, 37oc/30 s, 72oc/30 s). amplification products were resolved by electrophoresis on gel consisting of 1,5% agarose and tbe buffer with ethidium bromide 10 μl/ 100 ml. genetic similarities among genotypes were estimated using nei’s equation (n e i and l i 1979). results scots pine stand. among analysed 34 stumps, 23 were colonized by heterobasidion annosum s. s. in 80% stumps the pathogen was also present in root systems. on the base of somatic incompatibility test this area was colonized by 32 genets of h. annosum sensu stricto. genets were very small. one genet occupied mainly only one stump. in five stumps there were found two different genets and in one stump – three. genetic similarity of genets varied from 0% to 62%. (fig.1). the genetic diversity of the h. annosum sensu stricto population was high. only in 5% cases the genetic similarity between two genotypes was relatively high (40% – 62%) and in 1,5% was low (5%). in 24% cases the compared genets were genetically different. genetic similarity of genets occupying the same stump varied from 0% – 54%. in stump where three genets colonized wood similarity among genets was 10%, 18% and 40%. norway spruce stand. in this stand 21 stumps were located in the plot. 15 stumps (71%) were colonized by heterobasidion parviporum. 24 genets colonized this area. genets were small and occupied only one stump at least. in two cases, four different fig.1. dendrogram of similarity among genotypes of h. annosum sensu stricto. 206 p. łakomy et al. genets were found in stump, but in five cases 2 different genets colonized one stump. genetic similarity of genets varied from 0% to 38% (fig.2). in 4% cases genetic similarity between two genets varied from 30% – 38%. the lowest genetic similarity between genets was 9%. also in 9% genets were totally different. similarity of genets that occupied the same stump varied from 4% to 35%. in stumps colonized by 4 genets, their similarity varied from 16% to 35%. there were no genetically different genets in one stump. european silver fir stand. on the first plot heterobasidion abietinum colonized laying log and only two stumps (29%) and on the second the pathogen occurred in fig.3. dendrogram of similarity among genotypes of h. abietinum. fig.2. dendrogram of similarity among genotypes of h. parviporum. genetic diversity 207 3 tumps (60%). on the base of somatic incompatibility stumps were colonized by 14 genets (7 in each plot). in the laying log three genets were localized. two were small and occupied about 50 cm of stem. the third genet colonized wood on distance 7m of stem. three stumps were colonized by two genets in each stump and in one stump grew four genets of h. abietinum. one stump was whole colonized by one genet. genetic similarity of genets varied from 0% to 55% (fig. 3). the highest similarity (40% – 55%) was found in 5% of compared genets. in 13% cases the genets were genetically different. similarity among genets colonized the same stump varied from 0,16% to 36%. discussion in this study all genets localized in norway spruce and scots pine stands were small. the most probable infection mode was done by basidiospores after thinning. each stand was thinned at least three times (precommercial and commercial thinning). in scots pine stands only after the last thinning all stumps were treated against heterobasidion annosum s.s. with phlebiopsis gigantea (fr.: fr.) jülich., while stumps in the norway spruce stand had never been treated with biopreparation. there were good conditions for stump infection for a long time. both pine and spruce stand grew as a first generation of forest in post-arable soil. so the small size of genets might be resulted of relatively short time of heterobasidion spp. existence in those stands. although the vegetative spread of pathogens mycelia should be considered, even if genets were restricted to single stump. completely different situation was observed in a. alba stand. lack of stumps in this stand was connected with age of trees and very limited treatment. only dead or damaged by wind trees had been removed for last years. so almost all stumps were decayed. those genets probably were older than in pine and spruce stands. probably there were two reasons of small genet size – age of stumps and distance among stumps (2-15m). the root system around stumps had been decaying and diminishing for years and possible vegetative spread was also difficult. in addition size of genets colonizing the root system could also shrink. the proof of this idea is the size of genet in laying log. the stem was colonized after felling, because butt of stem was not colonized by h. abietinum. the biggest genet, which has been found, occupied wood on distance 7m. considered the speed of mycelium growth in dead wood (50 cm per year) this genet exists for at least 14 years. this observation improves earlier findings (k o w a l s k i , ł a k o m y 1998) that root pathogen was able to infect the higher parts of stem after tree felling. probably it was in connection with dead wood. k a l i o (1970) found that heterobasidion annosum spores could spread even up to 500 km. the high genetic diversity of pathogen’s genets and occurrence of many different or weakly similar genets could be explained by high pressure of spores from vicinity stands. this situation was also caused by lack of stump treatment against heterobasidion sp. after thinning. the age of norway spruce and scots pine stands influenced the size of genets. at the beginning of stands colonization the vegetative spread via root systems must be limited. the typical life span of heterobasidion spp. mycelium in roots is still not known. however on the base of the average rapid mycelial growth in roots the largest area 208 p. łakomy et al. occupied by a single genet was estimated on 50 m in diameter and the age of this genet would not be much more than 100 years (s t e n l i d and r e d f e r n 1998). in investigated stands genets were very young and the oldest one found in fir stand has less than 20 years. in coniferous stands a lot of h. annosum genotypes could exist in disease stand or gap (c h a s e , u r l i c h 1983; g a r b e l o t t o 1996; h a r r i n g t o n et al. 1998). -s t e n l i d et al. (1998) found that size of genotypes could be large and one genotype could colonize even 15 trees. however p i r i et al. (1990) and p i r i (1996) indicated that genotypes in investigated stands colonized only 3 trees. b o d l e s et al. (2005) de-b o d l e s et al. (2005) de-(2005) described genets in sitka spruce stand, which had been never thinned before. they found that maximum number of trees affected by one genet was 6 and genet size varied from those occupied single tree to 22,5 m in length. the age of the largest genets, which were 22,5 m in length, 17,5 m and 9 m, they estimate on 45, 35 and 18 years, respectively. s w e d j e m a r k and s t e n l i d (1993) suggested that even if tree was colonized by two or more genets only one may eventually dominate in a single host tree and wo o d w a r d et al. (2005) partially supported that hypothesis. ł a k o m y , b o r o d a and we r n e r (2005, unpublished) observed that in some cases only one genet from two or more, which colonized the same stump characterize of highest growth rate in wood of living tree. this situation was noted for h. annosum ss., h. parviporum and h. abietinum genets existing in one stump (pine, spruce or fir). but at the other hand they also observed two dominant genets among three or four from the same stump. it might be result of competition between genets in wood colonization. however they also find no differences in wood colonization among two or three genets from a single spruce or even small pine stump. previous works indicated on existing variation in virulence between heterobasidion species and also between isolates inside each species (we r n e r 1987, 1991; s t e n l i d , s w e d j e m a r k 1988; s w e d j e m a r k , s t e n l i d 1993; we r n e r , ł a k o m y 2001, 2002). ł a k o m y et al. (2005) suggested that damages might be related with number of active genets of heterobasidion in stand. they showed that the variation of virulence among isolates belonged to the same species might be important for disease progress in stands. in their experiments the highest mortality caused both genets of heterobasidion annosum genetically different and also in some degree similarity (30%). those three populations of h. annosum ss., h. parviporum and h. abietinum characterized of high genetic diversity. in pine and spruce stands this situation was resulted the high success in heterobasidion spp. colonization of fresh stumps appearing after routine thinning operation. those genets are young because both stands growth as a first generation on post agricultural soil. moreover in spruce stands forest service had never treated stump s against h. parviporum and in pine stand this operation had been done too late – during last commercial thinning. at least twice stumps after precommercial thinning were open to spore infection. different situation observed in fir stand was connected with trees’ age (180-years-old). some genets of h. abietinum must be old especially those which were present in old decayed stumps. in addition law number of stumps in stand was an important barrier to h. abietinum distribution. genetic diversity 209 acknowledgements: the authors are grateful to dr kari korhonen and prof. paolo capretti for providing pure cultures of heterobasidion spp. annosum and anna ratajczak, arleta świetlik, anna błaszkowiak, leokadia torz, ewa nowak for their assistance in this project. the polish committee for scientific research financially supported this study, grant no. 3p06l 054 22. references b o d l e s w. j. a., b e c k e t t l., z a m p o n i s., w o o d w a r d s., k e č a n., c a p r e t t i p. 2005. heterobasidion annosum population recruitment and spread in a severely infected sitka spruce stand in north east scotland. 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(in:) c. d e l a t o u r , j. j. g u i l l a u m i n , b. l u n g e s c a r m a n t , b. m a r c a i s (eds): root and butt rots of forest trees (9th international conference on root and butt rots), inra editions (france), les colloques no 89: 75–84. s w e d j e m a r k g., s t e n l i d j. 1993. population dynamics of the root rot fungus heterobasidion annosum following thinning of picea abies. oikos 66: 247–254. t h o m p s o n d., h e n r y r. 1995. single–step protocol for preparation of plant tissue for analysis by pcr. bioo. techniques 19: 394–400. va s i l i a u s k a s r., s t e n l i d j. 1998. spread of s and p group isolates of heterobasidion annosum within and among picea abies trees in central lithuania. can. j. for. res. 28: 961–966. we e b r. s., a l e x a n d e r s. a. 1985. an updated host index for heterobasidion annosum. virginia polytechnic inst. state university inform series no. 85–2: 27pp (abstr. in rev. plant pathol. 67: 63. 1988). we r n e r a. 1987. responses in vitro grown pine seedlings to infection by four strains of heterobasidion annosum. eur. j. for. path. 17: 93–101. we r n e r a. 1991. resistance to heterobasidion annosum of first and second generation scots pine trees from an old disease centre. arboretum kórnickie rocznik 36: 113–126. we r n e r a., łakomy p. 2001. intraspecific variation in heterobasidion annosum (fr.) bref. for mortality rate on pinus sylvestris l. and picea abies (l.) karst. seedlings grown in pure culture. mycologia 94 (5): 855–860. we r n e r , a., ł a k o m y p. 2002. host specialization of is–group isolates of heterobasidion annosum to scots pine, norway spruce and common fir in field inoculation experiments. dendrobiology 47: 59–68. w o o d w a r d s., h a r a m b u r u e., j o h n s t o n d. h. 2005. heterobasidion annosum in a severly affected norway spruce stand following chemical thinning. (in:) m. m a ń k a and p. ł a k o m y (eds). root and butt rots of forest trees. proceedings of iufro working party 7.02.01. 11th international conference on root and butt rots. poznań–białowieża, poland. 16–22 aug. 2004: 363–371. zróżnicowanie genetyczne grzybów rodzaju heterobasidion w drzewostanach: sosnowym, świerkowym i jodłowym s t r e s z c z e n i e badania nad zróżnicowaniem genetycznym gatunków rodzaju heterobasidion w drzewostanach sosnowym, świerkowym i jodłowym wskazały, że niemalże wszystkie genotypy występujące w badanych drzewostanach były małe i zasiedlały co najwyżej jeden pniak. w kilku przypadkach stwierdzono występowanie dwóch, trzech, a nawet czterech genotypów w jednym pniaku. podobieństwo genetyczne genotypów h. annosum s. s. wynosiło od 0% do 62%, h. parviporum od 0% do 38%, a h. abietinum od 0% do 55%. najstarszy i największy genotyp stwierdzono w leżącej kłodzie jodłowej, której drewno przerastał, przez co najmniej 14 lat. ten genotyp należał do h. abietinum. rozmiar genotypów był związany z intensywnością cięć pielęgnacyjnych w drzewostanach, rozprzestrzenianiem zarodników, wiekiem drzewostanów oraz konkurencją w zasiedlaniu drewna pomiędzy genotypami. 2014-01-01t11:46:17+0100 polish botanical society first records of arcyria marginoundulata nann.-bremek. & y. yamam. (myxomycetes) in poland anna ronikier1, piotr perz2 and piotr chachuła3 1institute of botany, polish academy of sciences, lubicz 46, pl-31-512 kraków, a.ronikier@botany.pl 2z. nałkowskiej 12, pl-57-300 kłodzko, uslugi@data.pl 3pieniński park narodowy, jagiellońska 107b, pl-34-450 krościenko nad dunajcem piotrekchacha@gmail.com ronikier a., perz p., chachuła p.: first records of arcyria marginoundulata nann.-bremek. & y. yamam. (myxomycetes) in poland. acta mycol. 48 (2): 279–285, 2013. a new to poland species of a myxomycete, arcyria marginoundulata, was found at two distant localities in the southern part of the country. polish specimens are typical and have all important features characteristic of the species: minute, grey, stipitate sporocarps, calyculus concentrically plicate at margin, spiny capillitium and small spores covered with few irregularly distributed larger warts. the species was found growing on alder female catkins. it seems that this substrate is specific for a. marginoundulata in europe. key words: arcyria globosa, arcyriaceae, the carpathians, mycetozoa, protozoa, the sudetes introduction the genus arcyria belongs to the order trichiales and family arcyriaceae within the class myxomycetes. it includes 50 species (lado 2005-2013). species of arcyria are characterized by usually stalked sporocarps covered by fugacious peridium remaining at the base as calyculus, pale spores and elastic capillitium forming a network (e.g. poulain et al. 2011a). twelve species of arcyria have been reported from poland by drozdowicz et al. (2003) and one more species, arcyria imperialis (g. lister) q. wang & yu li has recently been found (see panek, romański 2010, as hemitrichia imperialis g. lister). most of these species are common in the country and frequently reported. only arcyria minuta has been recorded on less than five localities in poland (tabacki 1977, but see drozdowicz et al. 2003, stojanowska, panek 2004; panek, romański 2010). most arcyria species occur on wood of coniferous or deciduous trees, but some of them can be found on other substrata, such as pine needles (a. annulifera, lado, pando 1997) or debris of monocotyledons (a. riparia, poulain et al. 2011a). arcyria acta mycologica vol. 48 (2): 279–285 2013 doi: 10.5586/am.2013.030 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 280 a. ronikier et al. marginoundulata is another species that grows on specific substrates: alder female catkins or inflorescences of other trees (poulain et al. 2011a, b). in the present paper we report this species, hitherto not known from poland, from two distant localities in the southern part of the country. material and methods material was collected in the field in 2007. observations and measurements of the morphological characters of mature sporophores were done under stereoscopic microscope nikon smz 1500. the total height, height and width of sporotheca of ten sporophores per collection were measured. observations of microscopic characters were made on material mounted in hoyer’s medium or water and measurements on material mounted in hoyer’s medium, under a light microscope nikon eclipse e-600, with a nomarski interference contrast. spore measurements of well developed specimens (30 per collection) were made under an oil-immersion objective (100×) and include ornamentation. specimens are kept in kram. localities of species in poland were mapped according to the atpol grid square system as used by wojewoda (2000). for a comparison of a. marginoundulata with a similar species, a collection of a. globosa schwein. from kram was examined. on the original herbarium label there is information: “ex herb w.g. farlow” and the specimen was collected “in involucris castaneorum”. neither locality nor a date of collection is noted on the herbarium label, only: “newton” (probably a collector name?) and: “haüfig” (kram m-102). results species description arcyria marginoundulata nann.-bremek. & y. yamam. figs 1, 2 sporophores scattered, sporocarpic (fig. 1a). sporocarps stipitate, total height 0.26–1.06 mm. sporotheca globose or slightly wider than high (mean height/width ratio 0.94), 0.08–0.50 mm high, 0.10–0.48 mm wide, first pale red-brownish, then pale greyish when mature. stalk long, 0.18–0.70 mm, one to four times longer than the sporotheca height, narrowly conical, longitudinally furrowed, pale greyish at apex, rusty-brown to yellow-brown towards the base (figs 1b–d), yellow-brown by transmitted light. peridium fugacious, remaining as a small calyculus at sporotheca base. calyculus greyish, pale yellowish by transmitted light, radially wrinkled, margin thickened, concentrically plicate (fig. 2a), surface almost smooth, with faint reticulum-like pattern observed under light microscope. capillitium dense, elastic, slightly expanding, reticulate, large-meshed, in reflected light pale grey, in transmitted light pale yellowish and with yellow to yellow-brown, round inclusions inside swellings when observed in water, hyaline in hoyer’s medium, attached to the calyculus first records of arcyria marginoundulata 281 surface in several points, threads 1–3 μm wide, with some swellings, densely ornamented with 1–3(4) μm long, spines in outer part of sporotheca, or with low warts connected by a faint reticulum in inner part of sporotheca or close to the calyculus (fig. 2c–d). spines conical when observed in hoyer’s medium, blunt or even slightly fig. 1. arcyria marginoundulata nann.-bremek. & y. yamam. coll. kram m-1584: a – group of sporophores on an alder female catkin; b – young sporocarp; c, d – mature sporocarps. scale bars: a = 5 mm; b–d = 500 μm. 282 a. ronikier et al. enlarged at apex when observed in water. spores pale grey in mass, hyaline by transmitted light, (6) 6.5–8 (9.5) μm, thin-walled, with loosely and irregularly distributed smaller and larger warts (fig. 2b). localities in poland: (1) the central sudetes, the góry bystrzyckie mts, the śnieżnik massif, about 25 km south from kłodzko town, domaszków village, 16°42’13.69”e, 50°12’53.45”n, alt. 446 m, at the bank of the domaszkowski potok stream, under alder tree, on alder female catkins, 22 july 2007, leg. piotr perz, kram m-1584; (2) the western carpathians, the beskid mały mts, surroundings of wadowice, about 2 km se from andrychów, inwałd – „wapiennik”, 19°23’22” e, 49°50’59,1” n, alt. 390 m, along a stream, among alnus glutinosa and urtica sp.; on alder (alnus glutinosa) female catkins, 26 aug. 2007 and 01 oct. 2007, leg. piotr chachuła, kram m-1583. notes: our specimens nicely fit the original description of arcyria marginoundulata, however they slightly differ in stalk colour and length; it is dark brown and about 3/4 to 4/5 of the total sporocarp height in the type specimen (nannenga-bremekamp, yamamoto 1983). also size of spines on capillitium seem to differ from those described from the type material to be about 1 μm high (nannenga-bremekamp, yamamoto 1983). we observed spines up to 4 μm high in major part of the capillitium, while lower spines (about 1 μm high) cover capillitium threads close to the calyculus (fig. 2c–d). fig. 2. arcyria marginoundulata nann.-bremek. & y. yamam. coll. kram m-1583: a – calyculus observed in hoyer’s medium; b – spores observed in water; c, d – capillitium observed in water (c – top view, d –median view), note different ornamentation type in parts of capillitium: low and reticulate or high and spiny. scale bar = 10 μm. first records of arcyria marginoundulata 283 we also noted differences in spine shape depending on madium: conical with a pointed apex when observed in hoyer’s medium and blunt or even slightly enlarged at apex when observed in water. the polish collections are very similar to those described and illustrated by liu et al. (2002) and poulain et al. (2011a, b). arcyria marginoundulata is one of several pale-coloured species from the genus. other taxa occurring in temperate europe differ among others in shorter stalk, larger sporothecae and occurrence of other substrata. one of the most similar to a. marginoundulata is a. globosa that is the most often reported from chestnut burrs (e.g., martin, alexopoulos 1969; poulain et al. 2011a) but it has also been registered in moist chamber cultures from alder catkins (adamonyté 2001). arcyria globosa differs in shorter stalk (about ½ of the total sporophore height), larger and more convex calyculus that is not thickened and undulate at margin but thin, radially wrinkled at base and minutely concentrically wrinkled above (fig. 3a). additionally, a. globosa differs in capillitium ornamentation composed of low spines. athough in some places the spines merges into reticulum (and then the capillitium ornamentation is similar to those of a. marginoundulata from the basal part), the most often they are arranged in right-handed spirals (fig. 3b). we found specimens of a. marginoundulata at two localities in southern poland (fig. 4). our first specimen was found by the second author on alder female catkins. then it was searched for by the third author several days later in randomly selected localities with alnus glutinosa stands and it was easily found. thus, the species is probably common in poland but overlooked due to its untypical habitat. although it was reported from various substrata: inflorescence of quercus serrata in japan (nannenga-bremekamp, yamamoto 1983), inflorescence of castanea crenata in japan (poulain et al. 2011a, b), leaf litter, miscanthus floridulus, pseudosasa japonica, cryptomeria japonica, fallen leaves in taiwan (liu et al. 2002) and ground litter and coarse woody debris in texas, usa (winsett, stephenson 2012), a. marginoundulata seems to be associated with alder female catkins in europe (see also müller, schulz 2010). at our two localities the species occurred exclusively on alder female catkins and it was absent from other litter elements present at the localities. if the species is a frequent inhabitant of this substrate it is probably widespread in poland, since the distribution of alnus glutinosa covers almost the whole area of the country (zając, zając 2001). fig. 3. arcyria globosa schwein. coll. kram m-102: a – calyculus observed in hoyer’s medium; b – capillitium observed in water. scale bars: a = 100 μm, b = 10 μm. 284 a. ronikier et al. acknowledgments. we thank anna drozdowicz for information about localities of arcyria imperialis, to michał ronikier and two reviewers for valuable comments to the manuscript, and to marian wysocki and jacek wieser for a contour map of poland. this work was partly financed through the statutory found of the w. szafer institute of botany, polish academy of sciences. references adamonyté g. 2001. myxomycetes of viešvilė strict nature reserve (sw lithuania). 2. moist chamber cultures. botanica lithuanica 7 (2): 179-191. drozdowicz a., ronikier a., stojanowska w., panek e. 2003. myxomycetes of poland. a checklist. (in:) z. mirek (ed.) biodiversity of poland. 10. w. szafer institute of botany, polish academy of sciences, kraków. lado c. 2005-2013. an online nomenclatural information system of eumycetozoa. http://www.eumycetozoa.com (2013-01-29). lado c., pando f. 1997. myxomycetes. i. ceratiomyxales, echinosteliales, liceales, trichiales. real jardín botánico de madrid and j. cramer, madrid, berlin, stuttgart. liu c.h., yang f.h., chang j.h. 2002. myxomycetes of taiwan xiv. three new records of trichiales. taiwania 47: 97-105. martin g.w., alexopoulos c.j. 1969. the myxomycetes.university of iowa press, iowa city. müller h., schulz w. 2010. myxomyceten an fruchtständen von alnus in thüringen. z. mykol. 76 (1): 75-82. fig. 4. distribution of polish localities of arcyria marginoundulata nann.-bremek. & y. yamam. first records of arcyria marginoundulata 285 nannenga-bremekamp n.e., yamamoto y. 1983. additions to the myxomycetes of japan. i. proc. kon. ned. akad. wetensch. c 86 (2): 207-241. panek e., romański m. 2010. śluzowce myxomycetes. (in:) l. krzysztofiak (ed.). śluzowce myxomycetes, grzyby fungi i mszaki bryophyta wigierskiego parku narodowego. przyroda wigierskiego parku narodowego, seria naukowa: 9–85. stowarzyszenie „ człowiek i przyroda”, suwałki. poulain m., meyer m., bozonnet j. 2011a. les myxomycètes. 1. guide de determination. fmbds, sevrier. poulain m., meyer m., bozonnet j. 2011b. les myxomycètes. 2. planches. fmbds, sevrier. stojanowska w., panek e. 2004. myxomycetes of the nature reserve near wałbrzych (sw poland). part i. list of taxa and quantitative analysis. acta mycol. 39 (1): 47-63. tabacki a.p. 1977. slime molds (myxomycetes) of silesia beech woods. acta biol. 3, pr. nauk. uśl. 175: 58-66. winsett k.e., stephenson s.l. 2012. an annotated checklist of the myxomycetes of the big thicket national preserve, texas. j. bot. res. ints. texas 6 (1): 287-298. wojewoda w. (ed.) 2000. atlas of the geographical distribution of fungi in poland. 1. w. szafer institute of botany, polish academy of sciences, kraków. zając m., zając a. (eds.) 2001. distribution atlas of vascular plants in poland. laboratory of computer chorology, institute of botany, jagiellonian university, kraków. pierwsze stanowiska arcyria marginoundulata nann.-bremek. & y. yamam. (myxomycetes) w polsce streszczenie nowy dla polski gatunek śluzowca, arcyria marginoundulata, charakteryzujący się białopopielatymi zarodniami niewielkich rozmiarów (do ok. 0,5 mm średnicy) i występowaniem na szyszkach olszy, został znaleziony po raz pierwszy w polsce, na dwóch stanowiskach w południowej części kraju: w sudetach (góry bystrzyckie) oraz w karpatach (beskid mały). w niniejszej pracy przedstawiono opis polskich okazów oraz ich stanowisk, a także porównano a. marginoundulata z najbardziej podobnym gatunkiem, a. globosa, który może występować na podobnym substracie. arcyria marginoundulata jest gatunkiem kosmopolitycznym i mimo że znajdowanym na różnych substratach, w europie wydaje się być przywiązany do szyszek alnus. możliwe, że gatunek jest pospolity w kraju, ale nieodnotowywany z powodu małych rozmiarów zarodni oraz występowania na specyficznym i rzadko badanym substracie. 2013-12-20t14:52:07+0100 polish botanical society contribution to morphology and ecology of polyporus rhizophilus janusz łuszczyński and bożena łuszczyńska department of botany, institute of biology, jan kochanowski university świętokrzyska 15, pl-25-406 kielce, jluszcz@ujk.kielce.pl łuszczyński j., łuszczyńska b.: contribution to morphology and ecology of polyporus rhizophilus. acta mycol. 45 (2): 151–156, 2010. the distribution and ecological conditions of the occurrence of polyporus rhizophilus in poland are discussed. the species was previously known from only one locality in dwikozy near sandomierz. further localities were found on the roots of stipa capillata in the skorocice reserve near busko zdrój and at an anthropogenic site among dactylis glomerata, poa annua and p. trivialis grasses in an urban park in końskie. the latter locality is noteworthy as it is situated outside the occurrence range of xerothermic grasslands in the wyżyna kieleckosandomierska upland. this locality is isolated ecologically from the two remaining polish localities situated in the xerothermic grasslands. key words: graminicolous fungi, polyporus rhizophilus, threatened fungi, steppe fungi introduction fungi of the genus polyporus fr. are saprobes or, less frequently, parasites. they inhabit plants with wooded shoots (mostly species of deciduous trees, less often coniferous trees), causing white rot of wood and, exceptionally, of grasses. a total of 59 widespread species, of which a large number are species typical of tropical areas, are known worldwide (domański et al. 1967; domański 1984; kirk et al. 2008). the genus is represented in poland by 11 species, the majority of which are widespread in the country (wojewoda 2003). four species: p. brumalis, p. ciliatus, p. squamosus and p. varius, are common, while other species are rare. polyporus rhizophilus is one of the rarest species, with interesting ecology and geographical distribution (łuszczyński, łuszczyńska 2009). acta mycologica vol. 45 (2): 151–156 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 152 j. łuszczyński and b. łuszczyńska material and methods the results are part of long-term mycocoenological research conducted by authors in the wyżyna małopolska upland between 1986 and 2008 and are supplemented by literature data (sałata 1977; wojewoda 2003). biocoenotic and habitat conditions of the occurrence of polyporus rhizophilus were characterised using phytosociological relevés and habitat descriptions. phytosociological relevés were conducted with the commonly used braun-blanquet method. the nomenclature of vascular plants follows mirek et al. (2002). microscopic characters of the fruit-bodies, such as spores, basidia and hyphae, were measured using a micrometric scale. the drawings of such elements were based on photographs taken with a light microscope. herbarium specimens were deposited in the fungarium of the department of nature and mathematics, jan kochanowski university of humanities and sciences, kielce. results polyporus rhizophilus (pat.) sacc. fig. 1 syll. fung. (abellini) 11: 82 (1895) – polyporaceae, polyporales, incertae sedis, agaricomycetes, basidiomycota, fungi (kirk et al. 2008). syn.: leucoporus rhizophilus (pat.) pat., melanopus rhizophilus pat., polyporellus rhizophilus (pat.) pilát. morphology. polyporus rhizophilus produces single fruit-bodies that consist of a pileus and a stem. it grows at the base of living grasses on which the fungus parasitizes. pileus from 5 to 10 mm in diam., sometimes larger, even up to 4 cm (domański et al. 1967), flat, slightly convex at centre. upper surface light beige, light ochre, with traces of fine squamules. pileus margin concolourous with the pileus, acute, flat or slightly incurved in dry specimens. hymenophore tubular. pores ellipsoidal-rhomboidal, more rounded and smaller at the margin, 2–3 per 1 mm, white or creamcoloured, subdecurrent. stem centric or slightly eccentric, suberose, elastic, thin, smooth, 1 to 2–5 mm in diam. and up to 3 cm long, white at top, becoming dull brown to blackish at base. hyphae dimitic, hyaline; generative hyphae thin-walled, 3-5(-7) μm in diam., septae with clamps; skeletal hyphae thick-walled, branched, without septae, 2-5 μm in diam. basidia 15-25 × 4.5-6 μm, with a basal clamp. spores ellipsoidal 6-10 × (2-) 3-4 μm (fig. 1). localities in poland. prior to this paper, polyporus rhizophilus was known only from one locality in dwikozy in the wyżyna sandomierska upland (geographic coordinates: 50°44’n and 21°47’e; atpol square ef 82), where it produced fruitbodies on the roots of stipa capillata in the sisymbrio-stipetum capillatae association (sałata 1977; wojewoda 2003). two new localities were found in the wyżyna contribution to morphology 153 małopolska upland: one in the town of końskie and one in the skorocice reserve near busko zdrój (fig. 2). the locality in końskie is situated in the ne part of an urban park (geographic co-ordinates: 51o12’ n and 20o25’ e; atpol square – ee 32). a phytosociological relevé was performed to examine the plot in which polyporus rhizophilus occurred (400 m2). the tree layer sparse, crown density up to 50%, the shrub layer absent, the cover of the herb layer 85%, the moss layer scarce. grass in the herb layer mown regularly; participation of few perennials. grass-mowing considerably hindered the identification of the trophic relationship between the fungus and the grass species. fig. 1. polyporus rhizophilus: a – basidia; b – spores; c – generative hyphae; d – skeletal hyphae (ktc 3296). scale bar = 10 μm. 154 j. łuszczyński and b. łuszczyńska fruit-bodies of polyporus rhizophilus were found on 10 sept. 2001 while the phytosociological relevé was performed on 17 may 2002. the composition of the phytocoenosis of the patch was as follows: trees acer saccharinum 1.2, quercus robur 3.2, q. rubra 2.3; herb layer: acer platanoides (c) +, aegopodium podagraria 1.2, chelidonium majus +.2, dactylis glomerata 1.2, festuca altissima 1.2, ficaria verna 1.2, geranium robertianum 1.2, geum urbanum +, glechoma hederacea 1.2, heracleum sibiricum +.2, impatiens parviflora +, lamium album +, moehringia trinervia +.2, myosotis sylvatica +.2, poa annua 1.2, p. trivialis 3.3, plantago major +.2, polygonum aviculare +, quercus robur (c) +, ranunculus acris 1.2, rumex obtusifolius +.2, stellaria media 1.2, taraxacum officinale 1.2, urtica dioica 1.2, veronica chamaedrys 1.2. the second new locality is situated in the rock-steppe skorocice reserve near busko zdrój (geographic co-ordinates: 50o24’ n, 20o39’ e; atpol square – fe 24). fruit-bodies were found on 15 june 2007 in the central part of the reserve in stipa capillata tufts in the sisymbrio-stipetum grassland on the sw wall of the gorge. description of the locality. sw exposure; incline 20o; herb layer cover 90 %; relevé surface: 25 m2, 15 june 2007. soil belongs to initial gypsum rendzinas. floristic composition: anthericum ramosum +, anthyllis vulneraria 1.2, artemisia campestris +, astragalus danicus 1.2, campanula sibirica +, carex humilis +.2, euphorbia cyparissias 1.1, euphrasia stricta +, festuca valesiaca 1.2, galium verum 1.2, gypsophila fastigiata 1.2, koeleria macrantha +, medicago falcata +.2, pimpinella saxifraga +, poa bulbosa var. vivipara 1.2, orthanta lutea +, oxytropis pilosa 1.2, potentilla arenaria +.2, sisymbrium polymorphum 1.2, stipa capillata 4.4, thymus marschallianus +.2. geographic distribution. polyporus rhizophilus is known mostly from scattered localities in central, southern and eastern europe as well as from northern africa and asia (rauschert 1962; kotlaba 1984). it was recently recorded in south america (silveira, wright 2005; gomes-silva, gibertoni 2009). polyporus cryptopus, a species with a very similar morphology, biology and ecological requirements, occurs on prairie grasses of the great plains in north america. however, relationships of phylogenetic affinity between polyporus rhizophilus and p. cryptopus are not clear and it is unknown if both are the same species or two different ones (gilbertson, ryvarden 1987; ryvarden, gilbertson 1994). the distribution of p. rhizophilus in europe and the history of recent records are given by kreisel (2006). it is a rare component fig. 2. distribution of polyporus rhizophilus in poland. contribution to morphology 155 of the mycobiota everywhere. polyporus rhizophilus is limited to continental and submediterranean thermophilous steppe grasslands and extrazonal xerothermic communities within its range (ryvarden, gilbertson 1994; leshan 2008). it occurs on roots of numerous grass species belonging to genera such as bothriochloa, calamagrostis, chrysopogon, cynodon, digitaria, elymus, festuca, poa and stipa (jülich 1984; ryvarden, gilbertson 1994). however, it has mostly been recorded on stipa and is usually thought to be connected with this genus. discussion polyporus rhizophilus usually occurs in steppe and xerothermic grasslands that develop in the continental climate. it is therefore interesting that the fungus was recorded in an urban park in the town of końskie outside the northern range limit of the species where conditions are different from its ecological requirements. the occurrence of p. rhizophilus at an anthropogenic locality in which grasses connected with xerothermic habitats are absent may suggest that other factors enabling the development of the fungus are responsible for it. global warming and climate continentalisation in some regions seem to play a role. the influence of climatic changes on the range extension of some fungal species in discussed by kreisel (2006). according to kreisel (l.c.), the geographic range of the species has considerably expanded northwards in the last few years, which may be related to global warming. polyporus rhizophilus remains either a rare or very rare component of the mycobiota worldwide despite this tendency. while conditions favourable for range broadening have been observed, serious threats to the species are also recorded. they result from adverse biocoenotic changes that have been taking place in xerothermic grasslands and transformations of the usage type. overgrowing and excessive burning of grassy xerothermic communities cause the displacement and disappearance of grasses on which polyporus rhizophilus grows and, consequently, the disappearance of its localities. threats to and protection of polyporus rhizophilus in poland are related to the development dynamics, existence and protection of xerothermic biocoenoses. polyporus rhizophilus is a very rare component of the mycobiota in poland and its natural localities must be actively protected in situ. it is included in the red list of fungi threatened in poland as endangered species (e) (wojewoda, ławrynowicz 2006). references domański s. 1984. bondarzewiaceae (bondarcewowate), fistulinaceae (ozorkowate), ganodermataceae (lakownicowate), polyporaceae (żagwiowate). 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(in:) l. frey (ed.). grass research. w. szafer institute of botany, polish academy of sciences, kraków, pp. 119–127. mirek z., piękoś-mirkowa, h., zając, a., zając, m. 2002. flowering plants and pteridophytes of poland – a checklist. (in:) z. mirek (ed.). biodiversity of poland 1. w. szafer institute of botany, polish academy of sciences, kraków, 442 pp. rauschert s. 1962. polyporus rhizophilus pat. ein für deutschland neuer steppenpilz. westfälische pilzbriefe 3 (4): 53-59. ryvarden l., gilbertson r. l. 1994. european polypores. part 2. meripilus–trametes. synopsis fungorum 7: 388–743. fungiflora, oslo. sałata b. 1977. dwa nowe dla flory polski gatunki grzybów wyższych. fragm. flor. geobot. 23: 423–427. silveira r. m. b., wright j. e. 2005. the taxonomy of echinochaete and polyporus s. str. in southern south america. mycotaxon 93: 1–59. wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków, 812 pp. wojewoda w., ławrynowicz m. 2006. red list of the macrofungi in poland. (in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. 3 ed.: 53–70. w. szafer institute of botany, polish academy of sciences. kraków. dane do morfologii i ekologii polyporus rhizophilus streszczenie polyporus rhizophilus należy do rzadkich grzybów związanych z trawami stepów i kserotermicznych muraw rozwijających się w klimacie kontynentalnym. na stanowiskach naturalnych notowany bywał głównie na korzeniach wielu gatunków traw, jednak najczęściej z rodzaju stipa. w polsce znany był do tej pory z jednego stanowiska w dwikozach koło sandomierza. kolejne jego stanowiska zostały odszukane w rezerwacie skorocice koło buska zdroju, na korzeniach stipa capillata oraz w końskich, w parku miejskim, na siedlisku antropogenicznym, wśród traw dactylis glomerata, poa annua i p. trivialis. interesującym jest stanowisko w końskich, leżące poza północną granicą zasięgu polyporus rhizophilus, poza obszarem występowania muraw kserotermicznych na wyżynie kielecko-sandomierskiej. występowanie tego gatunku grzyba na izolowanym stanowisku być może należy wiązać z globalnym zjawiskiem ocieplenia klimatu. na ten problem zwracał już uwagę kreisel (2006), który zalicza m.in. polyporus rhizophilus do grupy grzybów ciepłolubnych rozszerzających swój areał na północ w kontekście aktualnych zmian klimatycznych. 2014-01-01t11:51:04+0100 polish botanical society interesting collections of phytopathogenic fungi agata wołczańska department of botany and mycology, m. curie-skłodowska university akademicka 19, pl-20-033 lublin, agata.wolczanska@poczta.umcs.lublin.pl wołczańska a.: interesting collections of phytopathogenic fungi. acta mycol. 45 (1): 91–96, 2010. ascochyta chaerophylli bres. and ramularia vallisumbrosae cavara are reported as new in poland. passalora bupleuri (pass.) u. braun on anthriscus sylvestris (l.) hoffm. is a new fungus/host combination and ten other rare species are recorded from new localities in the country. key words: ascomycota, anamorphic fungi, new records, distribution introduction recently issued “a preliminary checklist of micromycetes in poland” (mułenko et al. 2008) contains information about 5969 fungal species reported from this country. it is a big amount, but on the map of poland there are still regions which can be called “white spots” – with no information about phytopathogenic fungi, especially about anamorphic fungi. the present paper provides new data about 13 species collected recently in poland. two of them are new for poland, one is found on a new host plant and others were noted only on a few localities till now. they were collected in various regions of poland, but most of the stands are situated in the south part. materials and methods the collected specimens were air dried and examined by means of standard light microscopy. slide preparations were stained with cotton blue in lactic acid and warmed. the publications of braun (1998), crous and braun (2003), mel’nik (2000), sałata (1974, 1979, 2002) and teterevnikova-babajan (1987) were used for identification. the species new for poland are briefly described, illustrated and discussed. the remaining acta mycologica vol. 45 (1): 91–96 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 92 a. wołczańska species are enumerated with full details of new collection(s) and information on their previous findings in poland. the nomenclature of host plants follow mirek et al. (2002) and the names of physico-geographical regions of poland follow kondracki (1998). all the collected specimens are deposited in the herbarium of the botany and mycology department of maria curie-skłodowska university in lublin (lbl) and w. szafer institute of botany of polish academy of sciences in kraków (kram). results and discussion species newly found in poland ascochyta chaerophylli bres. leaf spots small, circular, 4-6 mm in diameter, yellow-brownish with darker margin; pycnidia inconspicuous, immersed in the leaf tissue, with brown-yellowish wall, 100-140 μm in diameter; conidia cylindrical, rounded at both ends, 2-celled, slightly constricted at the septum, 12-14 × 3.5-4.0 μm (fig. 1f). specimen examined. on chaerophyllum hirsutum l.western carpathians, beskid niski mts: iwonicz zdrój health resort, slope of glorieta mt., roadside, 16 sept. 1993, leg. a. wołczańska, lbl m-10062. remarks. this species is new to poland. it was previously reported on chaerophyllum spp. in the former czechoslovakia, germany, lithuania and united kingdom (meľnik 2000). in addition, ascochyta chaerophylli has been reported on falcaria vulgaris bernh. and sanicula europaea l. in moravia in the czech republic (sałata 2002). ramularia vallisumbrosae cavara leaf spots elliptical, oblong, 10-25 × 4-8 mm, at first yellow-brownish, brown, green-brownish, later with light center; leaf spots surrounded by a green halo, well visible on fading leaves. caespituli amphigenous (figs 1a, b). conidiophores emerged from well developed stroma-like hyphal aggregations, 10-180 × 2.5-3 μm (fig. 1 c). conidia catenate, 2–4-celled, ellipsoid-ovoid to cylindrical, 14-44 × 2-4 μm (fig. 1 d). specimens examined. on leucojum vernum l. eastern carpathians, bieszczady mts: vicinity of muczne village, alnetum incanae, 1 may 2007, leg. m. piątek, kram f-46634; bereżki village, 4 may 2007, leg. m. piątek, kram f-46635. remarks. in his monograph, braun (1998) mentioned this species from poland. in the available literature as well as in polish herbaria i did not find any reports or voucher specimens of ramularia vallisumbrosae from the territory of the country. it cannot be excluded that braun (1998) studied specimen(s) from german or other herbaria. anyway, this is the first clearly documented report of ramularia vallisumbrosae from poland. general distribution of this species includes europe and north america, but on leucojum it was noted only in the ukraine (braun 1998) and austria (farr, rossman 2009). interesting collections 93 new localities of species already known from poland cercospora zebrina pass. specimen examined. on trifolium repens l.western carpathians, pogórze bukowskie foothills: rymanów town, meadow, 11 june 1994, leg. a. wołczańska, lbl m-10053. remarks. this species has been hitherto reported from poland only few times. on trifolium repens it was known from pojezierze łęczyńsko-włodawskie lakeland (mułenko 1988) and rogoźnica village (danilkiewicz 1987). other hosts of this species are: trifolium dubium sibth. and trifolium sp. (świderska-burek 2008a). cladosporium variabile (cooke) g. a. de vries specimen examined. on spinacia oleracea l. wyżyna lubelska upland: lublin city, 11 may 2004, leg. m. chmiel, lbl m-10055. remarks. cladosporium variabile has been hitherto reported in poland only from lubartów town (moesz 1920, 1926), but it is possibly common in regions where spinach is cultivated because the general distribution of this species is very broad. it was reported from many countries in asia, europe and north america (usa) (dugan et al. 2004). passalora bupleuri (pass.) u. braun specimen examined. on anthriscus sylvestris (l.) hoffm. western carpathians, pogórze bukowskie foothills: rymanów town, meadow, 11 june 1994, leg. a. wołczańska, lbl m-10056. remarks. anthriscus sylvestris is a new host for passalora bupleuri in poland; previously it was reported in this country on anthriscus nitida (wahlenb.) hazsl., chaero phyllum bulbosum l., ch. hirsutum l., ch. temulum l. and coriandrum sativum l. (świderska-burek 2008b). protomyces kreuthensis krieg. specimens examined. on aposeris foetida (l.) less. western carpathians, pogórze bukowskie foothills: zmysłówka village, dentario glandulosae-fagetum, 2 june 1991, leg. a. wołczańska, lbl m-8639; eastern carpathians, góry sanocko-turczańskie mts: turnica reserve, dentario glandulosae-fagetum, 5 may 2007, leg. m. piątek, kram f-46529. remarks. this species has been previously known only from one locality in the bieszczady mts (sałata 1979). ramularia crassiuscula (unger) u. braun specimen examined. on delphinium oxysepalum borbás & pax. western carpathians, tatra mts: kobylarzowy żleb gully in the czerwone wierchy massif, 7 aug. 2004, leg. j. piątek & m. piątek, kram f-46533. remarks. ramularia crassiuscula has been recently reported from the tatra national park on delphinium oxysepalum growing on ciemniak mt. (mułenko, wołczańska 2004). the present locality is also in the czerwone wierchy massif in the tatra national park, but within the distance about 1 km from the previously published station. 94 a. wołczańska ramularia libanotidis bubák specimen examined. on falcaria vulgaris bernh. wyżyna małopolska upland: zwierzy-zwierzyniec village, n of busko zdrój town, xerothermic grassland, 10 july 2004, leg. j. piątek & m. piątek, kram f-46535. remarks. this species has been so far reported from poland only twice. schroeter (1908) found it on falcaria vulgaris in dzierżysław town near głubczyce town, and wołczańska (2005) detected the species on pimpinella saxifraga l. in lipowica village near dukla town. septoria hydrocotyles desm. specimen examined. on hydrocotyle vulgaris l. pobrzeża południowobałtyckie littorals: near jastarnia town, meadow, 7 sept. 1999, leg. a. wołczańska, lbl m-10052. remarks. septoria hydrocotyles has been rarely reported from poland; it was known from surroundings of konotop town near zielona góra town (hellwig 1899), jezioro czarcie lake near goleniów town (kućmierz 1974), anieliny village (michalski 1982), pojezierze łęczyńsko-włodawskie lakeland (mułenko 1988), and from szewce village near janów lubelski town (romaszewska-sałata et al. 1997, romaszewska-sałata, wołczańska 1997). septoria melampyri strasser specimens examined. on melampyrum pratense l. pobrzeża południowobałtyckie littorals: between chałupy village and kuźnica station, pine forest, 10 sept. 1999, leg. a. wołczańska, lbl m-10058; between kuźnica station and jastarnia town, pine forest, 11 sept. 1999, leg. a. wołczańska, lbl m-10059. remarks. in poland, septoria melampyri has been hitherto known only from białowieża national park (mułenko 1996; faliński, mułenko 1997). septoria polemonii thüm. specimen examined. on polemonium caeruleum l. pojezierza południowobałtyckie littorals: torfowisko kopaniarze peat-bog, e of brodnica town, eutrophic mire, 10 sept. 2004, leg. j. piątek & m. piątek, kram f-46537. remarks. the only previous report of septoria polemonii from poland derives from one hundred years ago: diedicke (1915) found this species in kostrzyn town in october 1904. septoria villarsiae desm. specimen examined. on nymphoides peltata (s. g. gmel.) kuntze: brama krakowska gate: kraków city, botanical garden, 23 july 2004, leg. m. piątek, kram f-46536. remarks. this is the second record of septoria villarsiae in poland. it was previously reported by moesz (1926) from zagożdżon reserve and this locality is now probably situated in the kozienicki landscape park. taphrina rhizophora johanson specimen examined. on populus alba l. brama krakowska gate: kraków–nowa huta city, near łąki nowohuckie meadow, 24 apr. 2007, leg. m. piątek, kram f-46638 (fig. 1e). interesting collections 95 remarks. the species has been reported from only 5 localities in poland so far: murowana goślina town, ludwikowo village, usarzewo village, bartniki village, puławy town (sałata 1974). acknowledgements. i am very grateful to maria chmiel, marcin piątek, and jolanta piątek, for permission to publish their collections. the paper was partially supported by grant from the ministry of science and higher education (mnisw) no n/n304/172436. references braun u. 1998. a monograph of cercosporella, ramularia and allied genera (phytopathogenic hyphomycetes). vol. 2. ihw verlag, eching. crous p. w., braun u. 2003. mycosphaerella and its anamorphs: 1. names published in cercospora and passalora. cbs biodiversity series 1. centraalbureau voor schimmelcultures, utrecht. danilkiewicz m. 1987 parasitic fungi occurring on meadows and pastures in the krzna river valley. zesz. probl. post. nauk roln. 307: 91–104 (in polish with english and russian summary). diedicke h. 1915. kryptogamenflora der mark branderburg. bd. ix. pilze vii. sphaeropsideae, melanconieae. verlag von gebrüder borntraeger, leipzig. dugan f. m., schubert k., braun u. 2004. check-list of cladosporium names. schlechtendalia 11: 1–103. faliński b., mułenko w. (eds). 1997. cryptogamous plants in the forest communities of białowieża national park. ecological atlas (project crypto 4). phytocoenosis 9 (n.s.), supplementum cartographiae geobotanicae 7: 1–522. farr d.f., rossman a.y. 2009. fungal databases, systematic mycology and microbiology laboratory, ars, usda. retrieved november 29, 2009, from http://nt.ars-grin.gov/fungaldatabases/. hellwig t. 1899. florenbild der umgegend von kontopp im kreise grünberg in schlesien. allgemeine botanische zeitschrift 5: 140–142; 157–160. kondracki j. 1998. geografia regionalna polski. państwowe wydawnictwo naukowe, warszawa. kućmierz j. 1974. a contribution to the knowledge of fungal parasitic flora of western pomerania (northwestern poland). fragm. flor. geobot. 20: 271–275. meľnik v. a. 2000. key to the fungi of the genus ascochyta lib. mitteilungen aus biologischen bundesanstalt für landund forstwortschaft 379: 1–192. michalski a. 1982. parasitic fungi of noteć meadows and neighbouring areas adjacent on the stretch nakło – ujście. acta mycol. 18: 175–202. mirek z., piękoś-mirkowa h., zając a., zając m. 2002. flowering plants and pteridophytes of poland. a checklist. biodiversity of poland 1: 1–442. w. szafer institute of botany, polish academy of sciences, kraków. moesz g. 1920. beiträge zur kenntnis der pilzflora von polen. i. mitteilung. botanikai közlemények 18: 22–28, (6)–(13). moesz g. 1926. additamenta ad cognitionem fungorum poloniae. ii. magyar botanikai lapok 1/12: 25–39. mułenko w. 1988. the microscopic pathogenic fungi of the łęczna-włodawa lake district. ii. the list of species. acta mycol. 24: 125–171. mułenko w. 1996. parasitic microfungi and their hosts collected on the study area. plant pathogenic fungi. (in:) j. b. faliński, w. mułenko (eds). cryptogamous plants in the forest communities of białowieża national park (project crypto). phytocoenosis 8 (n. s.), archivum geobotanicum 6: 55–65. mułenko w., majewski t., ruszkiewicz-michalska m. (eds). 2008. a preliminary checklist of micromycetes in poland. biodiversity of poland 9: 1–752. w. szafer institute of botany, polish academy of sciences, kraków. mułenko w., wołczańska a. 2004. new collections of ramularia species (hyphomycetes) in poland. acta mycol. 39 (1): 13–17. romaszewska-sałata j., sałata b., wołczańska a. 1997. new and rare species of sphaeropsidales in the polish flora. ii. acta mycol. 32 (2): 293–301. romaszewska-sałata j., wołczańska a. 1997. mikroskopowe grzyby fitopatogeniczne parku krajobrazowego „lasy janowskie”. (in:) s. radwan, b. sałata, m. harasimiuk (eds). środowisko przyrodni-przyrodni96 a. wołczańska cze parku krajobrazowego „lasy janowskie”. wydawnictwo umcs, ar lublin, park krajobrazowy „lasy janowskie”, lublin: 57–63. sałata b. 1974. flora polska. grzyby (mycota) 6: ascomycetes, taphrinales. pwn, warszawa–kraków. sałata b. 1979. flora polska. grzyby (mycota) 12: ascomycetes, protomycetales. pwn, warszawa– kraków. sałata b. 2002. polskie gatunki grzybów mitosporowych z rodzaju ascochyta. wydawnictwo umcs, lublin. schroeter j. 1908 (1897). die pilze schlesien. (in:) f. cohn (ed.). kryptogamen-flora von schlesien 3(2). j. u. kern’s verlag, breslau. świderska-burek u. 2008a. cercospora, cercosporella, cercosporidium. (in:) w. mułenko, t. majewski, m. ruszkiewicz-michalska (eds). a preliminary checklist of micromycetes in poland. biodiversity of poland 9: 353–358. w. szafer institute of botany, polish academy of sciences, kraków. świderska-burek u. 2008b. passalora. (in:) w. mułenko, t. majewski, m. ruszkiewicz-michalska (eds). a preliminary checklist of micromycetes in poland. biodiversity of poland 9: 436–440. w. szafer institute of botany, polish academy of sciences, kraków. teterevnikova-babajan d. n. 1987. griby roda septoria in sssr. izdateľstvo akademii nauk armjanskoj ssr, erewan (in russian). wołczańska a. 2005. the ramularia species in poland. monogr. bot. 95: 1–154. nowe informacje o grzybach fitopatogenicznych w polsce streszczenie praca zawiera dane na temat występowania 13 gatunków grzybów workowych (w stadium anamorfy i teleomorfy). dwa z nich (ascochyta chaerophylli, ramularia vallisumbrosae) są nowe dla polski, passalora bupleuri została zebrana na nowym żywicielu (anthriscus sylvestris), a pozostałe znane są z nielicznych stanowisk. 2014-01-01t11:50:35+0100 polish botanical society potentially pathogenic fungi in the material collected by the specialist regional hospital, łódź magdalena rusiecka1 and grażyna lipowczan2 1department of mycology, university of łódź, banacha 12/16 pl-90-237 łódź, magdalena.r@tlen.pl 2laboratory of mycology and sexually transmitted diseases, biegański hospital kniaziewicza 1/5, pl-94-374 łódź rusiecka m, lipowczan g.: potentially pathogenic fungi in the material collected by the specialist regional hospital, łódź. acta mycol. 45 (2): 197–205, 2010. the mycobiota responsible for the development of pathological changes of the skin and its adnexa in patients presenting at the specialist regional hospital, łódź, with suspected superficial mycosis between 01 may 2003 and 30 april 2005 is analyzed. in total of 2144 isolations 39.96% were dermatophytes, 39.39% were yeast-like fungi and 20.65% were moulds. candida albicans was the most frequently diagnosed species in fallowed by trichophyton rubrum. key words: infections, dermatophytes, yeast-like fungi, moulds introduction as potentially pathogenic fungi commonly occur in the biosphere, people are exposed to them throughout their development while susceptibility to fungal infection is universal and concerns people of all ages (kurnatowska 1998). fewer than 200 of ca 100 000 fungal species described so far are causative agents of diseases in people (richardson, warnock 1995; ławrynowicz 2002; chodorowska 2008). the intensity of infections and toxic or allergic reactions varies while quantitative and qualitative changes are observed in the range of fungi pathogenic to humans depending mostly on the environment (richardson, warnock 1995). the following factors are considered to condition the changes: the level of urbanisation and industrialisation of an area, geographic location and climatic conditions. the fungal biota also changes over time (boliński et al. 2003; baran, szepietowski 1994; dynowska et al. 2004). that is why for many years around the world epidemiologic investigations are conducted (baran et al. 1993; venugopal, venugopal 1993; khosravi et al. 1994; korstanje, staats 1995; merlin et al. 1999; o´grady, sahn 1999; foster et al. 2004; dynowska et al. 2008). acta mycologica vol. 45 (2): 197–205 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 198 m. rusiecka and g. lipowczan the prevalence of mycoses, both superficial and systemic, has been growing rapidly in the last few years. about 40% of the world population are estimated to suffer from fungal infections (kaszuba et al. 1997; chodorowska 2008). superfical mycoses are an especially serious epidemiological, medical and social problem (jabłońska, chorzelski 1988; gliński et al. 2002) and are commonly considered to be diseases associated with civilisation (bojarski et al. 2001; macura 2004). while they do not pose an immediate risk to human life and are often trivialized, fungal infections can be exceptionally bothersome by negatively influencing everyday life. not only do they cause health problems but also the physical appearance adversely affected by the pathogen makes those suffering from mycoses selfconscious. patients often feel they are limited both in professional and private lives. the discomfort and embarrassment related to the altered body image frequently undermines confidence, leads to diminished social contacts and everyday activities, and, in extreme cases, even to depression (nowicki 1999). dermatophytes, yeast-like fungi and moulds are the etiologic factors of superfical mycoses (macura 1998; dobrowolska et al. 2002). traditional methods of identification of this pathogens are based on phenotypic features, which, on the one hand, can be influenced by environmental factors, and, on the other hand, are very time – consuming and inefficient. thus, molecular methods of identification, based on genotype are more and more frequently applied. they are fast and reliable and may supplement traditional methods in the future (bojarski et al. 2001; dobrowolska et al. 2008; dobrowolska, jaworski 2008; nawrot 2008 ). material and methods fungi identified in the clinical material (01 may 2003 – 30 april 2005) obtained from patients referred to the laboratory of mycology and sexually transmitted diseases, specialist regional hospital (wojewódzki szpital specjalistyczny im. dr. władysława biegańskiego), łódź, with suspected mycosis of the skin and/or its adnexa were analysed. the taxonomic range of the mycobiota was analysed using descriptions of the clinical cases diagnosed. each description contained the following data: the patient’s sex,• the patient’s age,• the site or sites on the body from which the material was collected for mycologi-• cal analysis, the result of the initial direct examination,• culture results (species or genus of the fungus).• only those infections in which a positive result was obtained in culture were analysed. the material identification was based on the works by de hoog and guarro (1995) as well as baran (1998). the article presents species or genus of the fungus depending on the patient’s sex. other factors will be presented and analyzed in the next articles. potentially pathogenic fungi 199 results a total of 5 514 (100%) patients, including 3 370 (61.12%) women and 2 144 (38.88%) men, were examined. positive results of mycological tests (direct preparations confirmed in culture) were obtained for 2 103 patients (38.14%) and negative results for 3 411 (61.86%) patients (tab. 1). monofocal isolations were performed for 1 587 (75.46%) patients while the material was collected from more than one site changed pathologically in 516 patients (24.54%). in the group of 3 370 (100%) women, positive results of mycological tests were obtained in 1 298 (38.52%) cases and negative results in 2 072 (61.48%) cases. monofocal isolations were performed for 990 (76.27%) female patients and multifocal isolations for 308 (23.73%) women. the total number of pathological foci with positive mycological results was 1 493. in the group of 2 144 (100%) men, positive mycological results were obtained in 805 (37.55%) cases and negative results in 1 339 (62.45%) cases. in the group with positive results, monofocal isolations were conducted for 597 (74.16%) male patients while the material was collected from more than one pathological site in 208 (25.84%) men. the total number of pathological foci with positive results was 938. of a total of 2 475 (100%) isolations in men and women, 989 (39.96%) isolates were dermatophytes, 975 (39.39%) were yeast-like fungi and 511 (20.65%) were moulds. table 1 patients presenting with suspected mycosis between 01 may 2003 and 30 april 2005 month no of patients examined for mycosis no of patients with mycosis diagnosed no of patients with no mycosis diagnosed n % n % v 2003 201 96 47.76 105 52.24 vi 2003 238 85 35.71 153 64.29 vii 2003 235 78 33.19 157 66.81 viii 2003 213 78 36.62 135 63.38 ix 2003 206 86 41.75 120 58.25 x 2003 195 69 35.38 126 64.62 xi 2003 187 54 28.88 133 71.12 xii 2003 151 43 28.48 108 71.52 i 2004 229 79 34.50 150 65.50 ii 2004 276 77 27.90 199 72.10 iii 2004 325 152 46.77 173 53.23 iv 2004 260 81 31.15 179 68.85 v 2004 212 93 43.87 119 56.13 vi 2004 274 123 44.89 151 55.11 vii 2004 107 44 41.12 63 58.88 viii 2004 248 106 42.74 142 57.26 ix 2004 231 98 42.42 133 57.58 x 2004 257 109 42.41 148 57.59 xi 2004 259 96 37.07 163 62.93 xii 2004 212 80 37.74 132 62.26 i 2005 240 95 39.58 145 60.42 ii 2005 232 75 32.33 157 67.67 iii 2005 259 101 39.00 158 61.00 iv 2005 267 105 39.33 162 60.67 total 5514 2103 38.14 3411 61.86 200 m. rusiecka and g. lipowczan candida albicans (robin) berkhout (847 isolations, 34.22%) was the most frequently diagnosed species, followed by trichophyton rubrum (castellani) sabouraud (586 isolations, 23.68%) (tab. 2). in the group of women (1 524 isolations 100%), 540 (35.43%) isolations were dermatophytes, 664 (43.57%) were yeast-like fungi and 320 (21.00%) were moulds (table 2). c. albicans (588 isolations, 38.58%) and t. rubrum (301 isolations, 19.75%) were the most frequently diagnosed species (tab. 2). table 2 aetiological factor depending on the patient’s sex species women men total n % n % n % t. rubrum 301 19.75 285 29.97 586 23.68 t. mentagrophytes var. granulosum 93 6.10 64 6.73 157 6.34 t. mentagrophytes var. interdigitale 2 0.21 2 0.08 t. tonsurans 11 0.72 10 1.05 21 0.85 m. canis 127 8.33 76 7.99 203 8.20 m. gypseum 1 0.11 1 0.04 e. floccosum 8 0.52 11 1.16 19 0.77 dermatophytes total 540 35.43 449 47.21 989 39.96 c. albicans 588 38.58 259 27.23 847 34.22 c. glabrata 7 0.46 3 0.32 10 0.40 c. krusei 3 0.20 1 0.11 4 0.16 g. candidum 39 2.56 27 2.84 66 2.67 rhodotorula sp. 8 0.52 6 0.63 14 0.57 trichosporon sp. 3 0.20 2 0.21 5 0.20 pityrosporum sp. 13 0.85 11 1.16 24 0.97 p. ovale 3 0.20 2 0.21 5 0.20 yeast-like fungi 664 43.57 311 32.70 975 39.39 s. brevicaulis 43 2.82 25 2.63 68 2.75 a. niger 30 1.97 16 1.68 46 1.86 a. fumigatus 23 1.51 15 1.58 38 1.54 aspergillus sp. 5 0.33 1 0.11 6 0.24 penicillium sp. 2 0.13 2 0.08 other moulds 217 14.24 134 14.09 351 14.18 moulds total 320 21.00 191 20.08 511 20.65 total 1524 100 951 100 2475 100 table 3 dermatophytes: species species women men total n % n % n % t. rubrum 301 55.74 285 63.47 586 59.25 t. mentagrophytes var. granulosum 93 17.22 64 14.25 157 15.87 t. mentagrophytes var. interdigitale 2 0.45 2 0.20 t. tonsurans 11 2.04 10 2.23 21 2.12 m. canis 127 23.52 76 16.93 203 20.53 m. gypseum 1 0.22 1 0.10 e. floccosum 8 1.48 11 2.45 19 1.92 dermatophytes total 540 100 449 100 989 100 potentially pathogenic fungi 201 in the group of men (951 isolations 100%), 449 (47.21%) isolations were dermatophytes, 311 (32.70%) were yeast-like fungi and 191 (20.08%) were moulds (tab. 2). t. rubrum (285 isolations, 29.97%) and c. albicans (259 isolations, 27.23%) were the most frequently diagnosed species (tab. 2). t. rubrum (586 isolations, 59.25%), followed by microsporum canis bodin (203 isolations, 20.53%) and t. mentagrophytes var. granulosum (robin) blanchard (157 isolations, 15.87%) was the most frequently diagnosed species of the 989 (100%) strains of dermatophytes isolated (tab. 3). of the 975 (100%) strains of yeast-like fungi, c. albicans (847 isolations, 86.87%), followed by geotrichum candidum link (66 isolations, 6.77%) and the genus pityrosporum (24 isolations, 2.46%), was the most frequently diagnosed fungus while pityrosporum ovale (0.51%) was isolated in five cases (tab. 4). in the group of the 511 (100%) strains of moulds, scopulariopsis brevicaulis ((sacc.) bainier 68 isolations, 13.31%), followed by aspergillus niger v. tieghem (46 isolations, 9.00%) and aspergillus fumigatus fresenius (38 isolations, 7.44%), was the most frequently identified species. pathogens were identified as a mould and further identification was not conducted in 351 (68.69%) cases (tab. 5). in the group of women, t. rubrum (301 isolations, 55.74%), followed by m. canis (127 isolations, 23.52%) and t. mentagrophytes var. granulosum (93 isolations, 17.22%), was the most frequently identified taxon of the 540 (100%) strains of dermatophytes isolated (tab. 3). of the 664 (100%) strains of yeast-like fungi, c. albicans (588 isolations, 88.55%) dominated, followed by g. candidum (39 isolations, 5.87%) (tab. 4). in the group of 320 (100%) strains of moulds isolated, s. brevicaulis (43 isolations, 13.44%), a. niger (30 isolations, 9.38%) and a. fumigatus (23 isolations, 7.19%) were the most frequently isolated species. pathogens were identified as a mould and further identification was not conducted in 217 (67.81%) cases (tab. 5). in the group of men, t. rubrum (285 isolations, 63.47%), followed by m. canis (76 isolations, 16.93%) and t. mentagrophytes var. granulosum (64 isolations, 14.25%), was the most frequently diagnosed species of the 449 (100%) strains of dermatophytes isolated (tab. 3). of the 311 (100%) strains of yeast-like fungi isolated, c. albicans (259 isolations, 83.28%) was definitely the most frequently isolated species, followed by g. candidum (27 isolations, 8.68%) (tab. 4). in the group of 191 (100%) strains of moulds, s. brevicaulis (25 isolations, 13.09%), a. niger (16 isolations, 8.38%) and a. fumigatus (15 isolations, 7.85%) were the most frequently table 4 yeast-like fungi: species species women men total n % n % n % c. albicans 588 88.55 259 83.28 847 86.87 c. glabrata 7 1.05 3 0.96 10 1.03 c. krusei 3 0.45 1 0.32 4 0.41 g. candidum 39 5.87 27 8.68 66 6.77 rhodotorula sp. 8 1.20 6 1.93 14 1.44 trichosporon sp. 3 0.45 2 0.64 5 0.51 pityrosporum sp. 13 1.96 11 3.54 24 2.46 p. ovale 3 0.45 2 0.64 5 0.51 yeast-like fungi total 664 100 311 100 975 100 202 m. rusiecka and g. lipowczan diagnosed species. pathogens were identified as a mould and further identification was not conducted in 134 (70.16%) cases (tab. 5). discussion fungal isolations confirmed in culture were recorded in 38.14% of the total group in the study period. although considerably more women (61.12%) than men (38.88%) were examined, positive results in both groups were recorded in fewer than 50% patients. this corresponds to both world and polish trends for superficial mycoses. a higher percentage of positive results (55.54%) was recorded in studies by kaszuba et al. (1997) comprising all mycological diagnostic data from the łódź region between 1982 and 1986. however, women always constituted a more numerous group (53.33%). positive results were recorded in only 32% of the total study group in studies by erkiert-polguj et al. (2008) and women were diagnosed more often than men. dermatophytes which constituted 39.96% of all mycological isolations were observed in only a small majority of infections, almost immediately followed by yeastlike fungi (39.39%). moulds constituted 20.65% isolations. similar results were observed by kaszuba et al. (1997), sikora et al. (2000) and boliński et al. (2003). nowicki et al. (2006) report that non-dermatophytic fungi were major pathogens causing superficial mycoses in studies conducted in the gdańsk region between 2003 and 2005. erkiert-polguj et al. (2008) demonstrate the dominance of infections caused by yeast-like fungi which constituted 54% of infections diagnosed while dermatophytic infections accounted for 40.5%. c. albicans (34.2%) was the most frequently isolated species in the present study, while the keratinophilous and keratinolytic t. rubrum constituted 23.7% of isolations. similar results: candida sp. (39.08% of all isolations) and t. mentagrophytes (27.90%), were obtained in studies conducted in the wrocław region between 1995 and 1999 (sikora et al. 2000). t. mentagrophytes var. granulosum (32.35% of total isolations) and t. rubrum (24.99%) were the most frequently isolated species in studies conducted on patients with mycoses of the skin and its adnexa treated at the outpatient clinic, city hospital, health care centre, białystok, between 1996 and 2001 (boliński et al. 2003). table 5 moulds: species species women men total n % n % n % s. brevicaulis 43 13.44 25 13.09 68 13.31 a. niger 30 9.38 16 8.38 46 9.00 a. fumigatus 23 7.19 15 7.85 38 7.44 aspergillus sp. 5 1.56 1 0.52 6 1.17 penicillium sp. 2 0.63 2 0.39 other moulds 217 67.81 134 70.16 351 68.69 moulds total 320 100 191 100 511 100 potentially pathogenic fungi 203 t. rubrum (59.25% of isolations) was the most frequently identified species of dermatophytes in the study period (01 may 2003 – 30 april 2005), followed by m. canis (20.53%) and t. mentagrophytes var. granulosum (15.87%). t. rubrum (34.15%) was also the most frequently isolated species in studies conducted in the łódź region between 1987 and 1996 while t. mentagrophytes var. granulosum constituted 23.03% and m. canis 11.82% of total isolations (kaszuba et al.1997). in their analysis of the geographic distribution of dermatophytes in poland between 1988 and 1992, baran and szepietowski (1994) also demonstrated that t. rubrum, a definite dominant throughout almost entire poland, was the most frequently recorded species, followed by t. mentagrophytes. sikora et al. 2000 report that t. mentagrophytes (62.7%) was the most frequently isolated dermatophytic fungus in the wrocław region between 1995 and 1999. t. rubrum (27.8%) was the second and epidermophyton floccosum (harz) langer. et milochevitch (6.2%) was the third most frequently identified species. these results closely correspond with the results obtained by erkiert-polguj et al. (2008) in the material collected by the clinical hospital of dermatology, medical university, łódź, between 2004 and 2006. the sequence of the three aetiological factors was the same: t. mentagrophytes, t. rubrum, e. floccosum. c. albicans (86.87%) was the most frequently diagnosed species of strains of yeast-like fungi in the study period (01 may 2003 – 30 april 2005). sikora et al. (2000) also report candida sp. as the genus dominant in this group of pathogens. s. brevicaulis (13.31%) was the most frequently diagnosed mould in the study period, followed by a. niger (9.00%) and a. fumigatus (7.44%). in 68.69% cases, pathogens were identified as moulds and further identification was not conducted. sikora et al. (2000) also show that s. brevicaulis was the most frequently identified species in this group of fungi. conclusions fungal isolations confirmed in culture were recorded in 38.14% of the total group in the study period and women were diagnosed more often than men. dermatophytes were observed in only a small majority of infections, almost immediately followed by yeast-like fungi. c. albicans was the most frequently diagnosed species in the present study, followed by t. rubrum. c. albicans was also the most frequently isolated species of strains of yeast-like fungi. t. rubrum was the most frequently isolated dermatophytic fungus and s. brevicaulis was the most frequently diagnosed mould. acknowledgements. we thank prof. dr hab. maria ławrynowicz (łódź) for valuable consultations during this study. the authors are indebted to prof. dr hab. maria dynowska (olsztyn) for comments on the manuscript and to the anonymous reviewer for suggestions on the text. 204 m. rusiecka and g. lipowczan references baran e., szepietowski j. 1994. rozmieszczenie geograficzne dermatofitów izolowanych ze zmian skórnych na terenie polski. mikologia lekarska 1 (1): 11–17. baran e. 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(in:) e. baran (ed.). mikologia – co nowego? cornetis, wrocław: 228–240. potentially pathogenic fungi 205 o´grady t. c., sahn e. e. 1999. investigation of asymptomatic tinea pedis in children. j. am. acad. dermatol. 24: 660–661. richardson m. d., warnock d. w. 1995. grzybice, rozpoznawanie i leczenie. springer, pwn, warszawa: 1–8; 55–57. sikora m., pachołek t., soter k., szepietowski j. 2000. analiza zakażeń grzybiczych skóry i jej przydatków w rejonie wrocławia w latach 1995 – 1999. mikologia lekarska 7 (3): 145–151. venugopal p. v., venugopal t. v. 1993. tinea capitis in saudi arabia. int. j. dermatol. 32: 39–40. grzyby potencjalnie chorobotwórcze w materiale z wojewódzkiego specjalistycznego szpitala w łodzi streszczenie zakażenia grzybicze, a szczególnie powierzchowne grzybice skóry, paznokci i włosów, stanowią obecnie poważny problem epidemiologiczny i terapeutyczny na całym świecie, a ich liczba nieustannie wzrasta. obserwowane zmiany dotyczą zarówno częstości jak i rodzaju grzybic i związane są z warunkami geograficznymi, klimatycznymi a także stopniem uprzemysłowienia i urbanizacji danego terenu. celem pracy była analiza wyników badań osób kierowanych w okresie 01.05.2003– 30.04.2005 do pracowni mikologiczno-wenerologicznej wojewódzkiego specjalistycznego szpitala im. dr wł. biegańskiego w łodzi z podejrzeniem zakażeń grzybiczych skóry i\lub jej przydatków. dokonano przeglądu mikobioty na podstawie opisu przypadków klinicznych. przebadano 5514 (100%) osób: 3370 (61,12%) kobiet i 2144 (38,88%) mężczyzn. dodatnie wyniki mikologiczne uzyskano u 2103 (38,14%) osób. spośród 2475 (100%) wszystkich izolacji u kobiet i mężczyzn łącznie, na dermatofity przypadało 989 (39,96%), grzyby drożdżoidalne 975 (39,39%), a na grzyby pleśniowe 511 (20,65%) izolacji. najliczniej izolowanymi gatunkami u wszystkich badanych były: candida albicans (34,22%) i trichophyton rubrum (23,68%). wśród dermatofitów najczęściej notowano t. rubrum (59,25%), znacznie rzadziej microsporum canis (20,53%) oraz t. mentagrophytes var. granulosum (15,87%). wśród grzybów drożdżoidalnych zdecydowanym dominantem był c. albicans (86,87%), a wśród grzybów pleśniowych scopulariopsis brevicaulis (13,31%). w przypadku 68,67% zakażeń identyfikację zakończono na zaliczeniu czynnika infekcyjnego do grupy grzybów pleśniowych. 2014-01-01t11:51:25+0100 polish botanical society epiphytic lichens of apple orchards in poland, slovakia, and italy daria zarabska1, anna guttová2, fabiana cristofolini3, paolo giordani4 and anna lackovičová2 1 natural history collection, faculty of biology, adam mickiewicz university, umultowska 89 pl-61-614 poznań, darzarabs12@tlen.pl 2 institute of botany, slovak academy of sciences, dúbravská cesta 14, sk-845 23 bratislava 3 iasma research and innovation centre, fondazione edmund mach-environment and natural resources area, via e. mach, i-38010 s. michele all’adige (trento) 4 botanic centre hanbury, university of genova, corso dogali 1m, i-16146 genova zarabska d., guttová a., cristofolini f., giordani p., lackovičová a.: epiphytic lichens of apple orchards in poland, slovakia, and italy. acta mycol. 44 (2): 151–163, 2009. following the study of epiphytic lichens in 30 apple orchards from poland, slovakia and italy the list of 74 taxa was prepared. the most common are the mesoto xerophytic and heliophilous species. the highest number of taxa was observed in slovak orchards. moreover, lichens shared with at least one other country were also noted mainly in slovakia. bark of apple trees seems to create favourable habitats for bacidia rubella, which together with strangospora pinicola were valuable founds in polish orchards. in slovak orchards, special attention should be paid to acrocordia gemmata, melanelixia glabra and usnea hirta. among interesting records in italian orchards, phaeophyscia hispidula and ph. kairamoi can be mentioned. key words: lichens, culture landscape, fruit trees, wielkopolska province, trentino region, western carpathians introduction in accordance with the assumption of sustainable development policy, the conservation of old orchards can play an important role in supporting the wide variety of wildlife. the presence of old orchards in agricultural land can rise species diversity (cf. jermaczek, jermaczek 2003). moreover, orchards as an example of culture landscape, can influence the spreading of some species (szczepańska 2008). until now, some lichenological inventories were conducted in the orchards (e.g., suza 1936a; halicz, kuziel 1958; kuziel 1964; kiss 1982; mészáros et al. 1984; czyżewska 1998; bartók 1999; kiszka 1999; lipnicki 2003; fałtynowicz et al. 2004; kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009). among flagship acta mycologica vol. 44 (2): 151–163 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 152 d. zarabska et al. examples of research focused on the investigation of biota in the fruit trees should be distinguish studies carried in czechia (majeriková-hlaváčková 1957), great britain (lush et al. 2005), hungary (mészáros et al. 1984), poland (kuziel 1964; lipnicki, sobieralska 2009) and romania (bartók 1999). records coming from studies of lichens only in apple orchards are scarce (mészáros et al. 1984; bartók 1999). more information is given from fruit trees level without direct reference to investigation of orchards (e.g., endlicher 1830; zahlbruckner 1894; suza 1936a, b; rydzak 1970; toborowicz 1976; lipnicki 1982; kiszka, piórecki 1991, 1994; lackovičová 1997; czyżewska 1998; śliwa 1998; lisická 1999; czarnota 2000; suppan, mayrhofer 2002; zalewska et al. 2004; guttová 2005; aptroot et al. 2006; john 2006; pišút et al. 2007; szymczyk, zalewska 2008). the aim of this research was to study the lichen species in a traditional apple orchards in the wielkopolska province (western poland), western carpathians (western and central slovakia), and trentino region (northern italy). list of currently recorded species and a review of older data are treated in this paper. study area studied orchards in poland were chosen on the border of równina nowotomyska plain and wał lwówecko-rakoniewicki rampart being a part of wielkopolska province (kondracki 2000). most of orchards investigated here were surrounded by crops and pasture land. mainly they laid in the neighborhood of farms. region represent typical lowland character situated in the agricultural landscape. the altitude range creates on the level of 70–100 m a.s.l. the annual average temperature is 8.3°c and the average annual precipitation is ca 550 mm (averages from years 1971–2000; acc. to farata 2004). the prevailing winds come from w, sw and nw. localities; 04-05. 2008: 1. władysławowo village near wąsowo, alt. 114 m, 52°23.44’n, 16°14.30’e; 2. wytomyśl−lipka mała village near lwówek town, alt. 127 m, 52°23.37’n, 16°11.07’e; 3. lipka wielka village near lwówek town, alt. 120 m, 52°24.03’n, 16°11.05’e; 4. wytomyśl lipka mała village near lwówek town, alt. 127 m, 52°23.40’n, 16°11.10’e; 5. wytomyśl village near nowy tomyśl town, alt. 101 m, 52°23.0’n, 16°10.36’e; 6. kozie laski village near nowy tomyśl town, alt. 96 m, 52°21.38’n, 16°10.15’e; 7. lipka wielka village near lwówek town, alt. 121 m, 52°24.17’n, 16°11.14’e; 8. wytomyśl village near nowy tomyśl town, alt. 105 m, 52°22.37’n, 16°10.58’e; 9. wytomyśl village near nowy tomyśl town, alt. 106 m, 52°22.37’n, 16°10.58’e; 10. wytomyśl village near nowy tomyśl town, alt. 107 m, 52°22.35’n, 16°10.33’e. all slovak orchards were localized in western carpathians mts., namely in malé karpaty mts., biele karpaty mts., štiavnické vrchy mts., nízke tatry mts., and stolické vrchy mts. the sites are situated from 160 m (bratislava, malé karpaty mts.) to 730 m a.s.l. (hybe, nízke tatry mts.). orchards were located in cadasters of the villages (city), in the vicinity of farms/housing estates, or were surrounded by pasture land and in the neighborhood of forest. the western carpathians are characterized by mountain climate with clearly marked vertical zonation (bielczyk et al. 2004). annual precipitations range from 600 to 800-1200 mm. epiphytic lichens of apple orchards 153 localities; 06.2008: 1. biele karpaty mts., poriadie-durcovci village near stará turá town, alt. 380 m, 48°46.34’n, 17°37.33’e; 2. malé karpaty mts., bratislava, beside the road to záhorská bystrica vil-malé karpaty mts., bratislava, beside the road to záhorská bystrica village, alt. 160 m, 48°13.11’n, 17° 02.02’e; 3. biele karpaty mts., adamovské kochanovce village, locality vinohrady, alt. 250 m, 48°52.23’n, 17°54.59’e; 4. biele karpaty mts., adamovské kochanovce village, alt. 220 m, 48°51.09’n, 17°55.05’e; 5. nízke tatry mts., hybe, near the saw mill in n part of village, alt. 730 m, 49°03.15’n, 19°50.10’e; 6. stolické vrchy mts., revúca, nw part of the town, ca 500 m from the railway, alt. 320 m, 48°41.20’n, 20°06.30’e; 7. biele karpaty mts., trenčín-orechové, n part of the village by orechový potok stream, alt. 350 m, 48°54.2’n, 18°02.15’e; 8. biele karpaty mts., nová bošáca village, grúň mt., alt. 550 m, 48°53.43’n, 17°47.42’e; 9. štiavnické vrchy mts., pukanec village, alt. 354 m, 48°22.08’n, 18°44.02’e; 10. štiavnické vrchy mts., pukanec-majere, locality teplá voda, alt. 370 m, 48°22.16’n, 18°45.02’e. italian apple orchards were selected in the non valley (trentino region – n italy). the non valley takes shape in western trentino, along the shores of the noce stream from the rocchetta ravin up to the palade and mendola pass, between 500 and 1400 m a.s.l. the valley is characterized by an intensive agricultural landscape, with a strong prevalence of apple orchards, which produce up to 15% of the italian production. the climate is alpine, with average yearly precipitation between 500 and 1200 mm and average yearly temperature about 9°c. localities; 03-04.2009: 1. near the santa giustina lake, alt. 530 m, 46°21.52’n, 11°03.30’e; 2. near the santa giustina lake, along the road, alt. 540 m, 46°21.52’n, 11°03.27’e; 3. near the santa giustina lake, alt. 595 m, 46°20.45’n, 11°03.23’e; 4. near sarnonico, alt. 970 m, 46°25.17’n, 11°08.34’e; 5. sarnonico, along the road to seio, alt. 970 m, 46°25.17’n, 11°08.34’e; 6. sarnonico, along the road to seio, alt. 970 m, 46°25.11’n, 11°07.58’e; 7. sarnonico, along the road to seio, alt. 960 m, 46°25.11’n, 11°07.55’e; 8. sarnonico, along the road to cavareno, alt. 1000 m, 46°24.44’n, 11°08.29’e; 9. near the santa giustina lake, along the road to sanzeno, alt. 590 m, 46°21.45’n, 11°03.50’e; 10. near the santa giustina lake, along the road to sanzeno, alt. 580 m, 46°21.42’n, 11°03.42’e. material and methods in every orchard one sample plot (20 × 50 m) in direction north-south was established. from each stand lichens were recorded carefully from the bark of apple (malus sp.) up to 1.6 m. nomenclature of lichens follows nimis and martellos (2008). voucher specimen are available at the poz herbarium. results and discussion in total 74 taxa were recorded in the investigated orchards of three countries (tab.1). similar results (in species number) were obtained by kościelniak (2004). specific circumstances prevailing in the orchards can have impact on occurrence of some species. apple orchards in comparison with other fruit plantations are characterized by quite good light conditions and difficult access to the water on the trunk (kuziel 1964). moreover, favorable conditions for some lichens can be also created by bark ph of malus sp. which is naturally higher in comparison with some other trees, e.g., 154 d. zarabska et al. ta bl e 1 e pi ph yt ic li ch en d iv er si ty in o rc ha rd s of p ol an d, s lo va ki a, a nd i ta ly sp ec ie s po la nd sl ov ak ia it al y n um be r of o rc ha rd s 1 2 3 4 5 6 7 8 9 10 1 2 3 4 5 6 7 8 9 10 1 2 3 4 5 6 7 8 9 10 p hy sc ia a ds ce nd en s (f r. ) h . o liv ie r + + + + + + + + + + + + + + + + + + + + + + + + + + + + . + p ha eo ph ys ci a or bi cu la ri s (n ec k. ) m ob er g + + + + + + + + + . + + + + + + + + + + + + + + + + + + + + p hy sc ia te ne lla ( sc op .) d c . + + + + + + + . + + + + . + + + . + + + + + + + + + + + + + c an de la ri el la x an th os tig m a (a ch .) l et ta u + + + . + + + + + + + + + + + + + + + + + + + . . + . + + . p ha eo ph ys ci a ni gr ic an s (f lö rk e) m ob er g + + . . + + + . . . + + + + + + + + + + + + + + + + + + + + x an th or ia p ar ie tin a (l .) t h. f r. + + + . + . . . . . + + + + + + + . + + + + + + + + + + . + c an de la ri el la re fle xa ( n yl .) l et ta u + + + + + + + . . . + + + + + + + + + + + + . . + . . . . . x an th or ia p ol yc ar pa ( h of fm .) r ie be r + + + + + . + . . + . + . . + + . . . + . + + + + + + . . . l ec an or a ha ge ni i ( a ch .) a ch . + + + . + . . . + . + + + + + + . . . . + . + . . . . . . + x an th or ia c an de la ri a (l .) t h. f r. . + + + . . . . . . . + . . + . . . . . + . . + + + + + . . p hy sc on ia g ri se a (l am .) p oe lt . + . . . . . . + . . . + + . . + + + + . + . + + . . . . . l ec an or a ex pa lle ns a ch . . . + . + + + . + + . . . . . . . . + + + + . . . . . . . . m el an oh al ea e xa sp er at ul a (n yl .) o . b la nc o et a l. + . + . + . + . . . . . . . . . . . + + . . . . . + + . . . c al op la ca c er in a (h ed w .) t h. f r. . + . . . . . . . . . . . . + + + . . . . . . + . . . + . . h yp oc en om yc e sc al ar is ( a ch .) m . c ho is y . . + . . . . . . + + . . . . . . . . . + + . . . . . . . . a m an di ne a pu nc ta ta ( h of fm .) c op pi ns & s ch ei d. + + + . + + + + + . + + + + + + + + + + . . . . . + . . . . pa rm el ia s ul ca ta t ay lo r + + + + + + + . . . . + + + + + . + + + . . . . . . . . . . h yp og ym ni a ph ys od es ( l .) n yl . + . + + + + . . . + + + . + + + . + + . . . . . . . . . . . l ec an or a co ni za eo id es c ro m b. . + + + + + + + + + . + . . . . + . + + . . . . . . . . . . l ec an or a ch la ro te ra n yl . + + + + . . + . . . + + . . + + . . + . . . . . . + . . . . l ep ra ri a in ca na ( l .) a ch . + . + . + + + + + + + . + . . . . . + . . . . . . . . . . . c la do ni a fim br ia ta ( l .) f r. . + + . . + . . . + . . . . . . . . . . . . . . . . . . . . m el an el ix ia fu lig in os a su bs p. fu lig in os a (d ub y) o . b la nc o et a l. . . . . . . . . . . . + + + + + . . + + . . . . . . . . . . p hy sc on ia e nt er ox an th a (n yl .) p oe lt . . . . . . . . . . . . + . . . . + + + . . . . . . . . . . sc ol ic io sp or um c hl or oc oc cu m ( st en h. ) v ěz da . . . . . . . . . . . . . . . + . + + + . . . . . . . . . . h yp og ym ni a tu bu lo sa ( sc ha er .) h av . . . . . . . . . . . + . . . . . . . + + . . . . . . . . . . p hy sc on ia d et er sa ( n yl .) p oe lt . . . . . . . . . . . . . . . . . + + + . . . . . . . . . . c an de la ri a co nc ol or ( d ic ks .) s te in . . . . . . . . . . . . . . . . . . . . + + + + + + + + + + l ec an ia c yr te lla ( a ch .) t h. f r. . . . . . + . . + . + + + . + + . . . . . . . + . . . + . + p ha eo ph ys ci a ch lo an th a (a ch .) m ob er g . . . . . . . . . . . . . . . . . . . . + + + + + + + + + + p ha eo ph ys ci a ka ir am oi ( v ai n. ) m ob er g . . . . . . . . . . . . . . . . . . . . + + + + + + + + + . x an th or ia fu lv a (h of fm .) p oe lt & p et ut sc hn ig . . . . . . . . . . . . . . . . . . . . + + + + + + + + . + p ha eo ph ys ci a hi rs ut a (m er es ch k. ) m ob er g . . . . . . . . . . . . . . . . . . . . + + + + . + + + + + p hy sc ia s te lla ri s (l .) n yl . . . . . . . . . . . . + . . + + . . . + . . . + + . . . . . epiphytic lichens of apple orchards 155 pa rm el in a til ia ce a (h of fm .) h al e . . . . . . . . . . . . . . + . . + + + . . . . . + . + . . m el an el ix ia s ub ar ge nt ife ra ( n yl .) o . b la nc o et a l. . . . . . . . . . . . . . . . . . + . . + . . . . + + + . . pe rt us ar ia a lb es ce ns ( h ud s. ) m . c ho is y & w er ne r . . . . . . . . . . . . . . . . . + . . . . . + . + + + . . p hl yc tis a rg en a (s pr en g. ) f lo t. . . . . . . . . . . . . . . . + . + + . . . . . . . . + . . m el an el ix ia g la br a (s ch ae r. ) o . b la nc o et a l. . . . . . . . . . . . . . . . . . . + + . . . . . + + . . . l ec id el la e la eo ch ro m a (a ch .) m . c ho is y . . . . . . . . . . . . . . + + . . . . . . . . . + . . . . m ic ar ea p ra si na f r. . . . . . . . . . . . . . . . . . . + + . . . . . . . . . + p hy sc on ia p er is id io sa ( e ri ch se n) m ob er g + . . . . . . . . . . . + . . . . . . . . . . . . . . . . . l ec an or a ca rp in ea ( l .) v ai n. . . + . . . . . . . . . . . . . . . + . . . . . . . . . . . d im er el la p in et i ( a ch .) v ěz da . . . . . + . . . . . + . . . . . . . . . . . . . . . . . . c la do ni a m er oc hl or op ha ea a sa hi na . . . . . + . . . . . . . . . . . . . . . . + . . . . . . . l ec an or a sy m m ic ta ( a ch .) a ch . . . . . . . + . . . . . . . . + . . . . . . . . . . . . . . m el an el ix ia s ub au ri fe ra ( n yl .) o . b la nc o et a l. . . . . . . . . . . . . . . + . . . + . . . . . . . . . . . p hy sc on ia d is to rt a (w it h. ) j. r . l au nd on . . . . . . . . . . . . . . . . . + + . . . . . . . . . . . a cr oc or di a ge m m at a (a ch .) a . m as sa l. . . . . . . . . . . . . . . . . . . + + . . . . . . . . . . p ha eo ph ys ci a hi sp id ul a (a ch .) e ss l. . . . . . . . . . . . . . . . . . . . . + . . . . . . + . . p ha eo ph ys ci a ce rn oh or sk yi ( n ád v. ) e ss l. . . . . . . . . . . . . . . . . . . . . . + . . . . . . . + p hy sc ia le pt al ea ( a ch .) d c . . . . . . . . . . . . . . . . . . . . . . . . + . . . + . . st ra ng os po ra p in ic ol a (a . m as sa l.) k ör b. . . + . . . . . . . . . . . . . . . . . . . . . . . . . . . o pe gr ap ha v ar ia p er s. . . . . . + . . . . . . . . . . . . . . . . . . . . . . . . c la do ni a co ni oc ra ea ( f lö rk e) s pr en g. . . . . . + . . . . . . . . . . . . . . . . . . . . . . . . b ac id ia ru be lla ( h of fm .) a . m as sa l. . . . . . . + . . . . . . . . . . . . . . . . . . . . . . . b ac id in a ar no ld ia na ( k ör b. ) v . w ir th & v ěz da . . . . . . . . . . + . . . . . . . . . . . . . . . . . . . m el an oh al ea e le ga nt ul a (z ah lb r. ) o . b la nc o et a l. . . . . . . . . . . + . . . . . . . . . . . . . . . . . . . f la vo pa rm el ia c ap er at a (l .) h al e . . . . . . . . . . . + . . . . . . . . . . . . . . . . . . l ec an ia n ae ge lii ( h ep p) d ie de ri ch & v an d en b oo m . . . . . . . . . . . + . . . . . . . . . . . . . . . . . . l ec an or a sa lig na ( sc hr ad .) z ah lb r. . . . . . . . . . . . + . . . . . . . . . . . . . . . . . . u sn ea h ir ta ( l .) f . h . w ig g. . . . . . . . . . . . + . . . . . . . . . . . . . . . . . . ps eu de ve rn ia fu rf ur ac ea ( l .) z op f . . . . . . . . . . . . . . + . . . . . . . . . . . . . . . pa rm el ia s ax at ili s (l .) a ch . . . . . . . . . . . . . . . + . . . . . . . . . . . . . . . c al op la ca h ol oc ar pa ( a ch .) a . e . w ad e . . . . . . . . . . . . . . . . . + . . . . . . . . . . . . e ve rn ia p ru na st ri ( l .) a ch . . . . . . . . . . . . . . . . . . . . + . . . . . . . . . . pe rt us ar ia a m ar a (a ch .) n yl . . . . . . . . . . . . . . . . . . . . . + . . . . . . . . . c al op la ca c itr in a (h of fm .) t h. f r. . . . . . . . . . . . . . . . . . . . . . + . . . . . . . . p hy sc ia d ub ia ( h of fm .) l et ta u . . . . . . . . . . . . . . . . . . . . . . . . + . . . . . l ec an or a al be lla ( pe rs .) a ch . . . . . . . . . . . . . . . . . . . . . . . . . + . . . . x an th or ia fa lla x (h ep p) a rn ol d . . . . . . . . . . . . . . . . . . . . . . . . . + . . . . a rt ho ni a ra di at a (p er s. ) a ch . . . . . . . . . . . . . . . . . . . . . . . . . . . . + . . m el an el ix ia fu lig in os a su bs p. g la br at ul a (d ub y) o . b la nc o et a l. . . . . . . . . . . . . . . . . . . . . . . . . . . . + . . o ch ro le ch ia a nd ro gy na ( h of fm .) a rn ol d . . . . . . . . . . . . . . . . . . . . . . . . . . . + . . to ta l n um be r of ta xa 74 16 17 21 10 16 18 16 6 11 10 17 23 15 13 23 21 10 19 29 25 19 18 14 18 15 23 16 23 8 13 156 d. zarabska et al. oak (svoboda 2007), and can promote occurrence of some specific, like nitrophytic, group of lichens. most of recorded in each country lichens have high indicator value for light (wirth 2001), e.g., caloplaca cerina, candelariella xanthostigma, cladonia fimbriata, hypogymnia physodes, lecanora conizaeoides, melanohalea exasperatula, parmelia sulcata, phaeophyscia nigricans, ph. orbicularis, physcia adscendens, ph. tenella, physconia enteroxantha, ph. grisea, xanthoria candelaria, x. parietina and x. polycarpa. some species − candelariella xanthostigma, phaeophyscia nigricans, ph. orbicularis, physcia adscendens, ph. tenella, xanthoria parietina and x. polycarpa represent mesophytic or xerophytic character (nimis, martellos 2008). most richest in species were orchards in slovakia (52). higher species richness can be partially explained by wider altitudinal range (dietrich, scheidegger 1997). less records were made in polish (32) and italian (43) orchards. climatic condi-climatic conditions, especially rainfall (lush et al. 2005) is one of the main factor impacted diversity of observed lichens in the particular countries. in region of poland where studies were conducted one of the smallest amount of rainfall (farata 2004) in comparison with other parties of poland as well as with studied sites in slovakia and italy can be observed. such climatic conditions surely negatively influenced growing of lichens. most of foliose broad-lichens represent montane elements in the achieved list of orchards lichens. some of narrow-lobed lichens (e.g., phaeophyscia chloantha and ph. hirsuta), recorded in alpine region of trentino represent submediterranean element in the lichen biota. nevertheless such species like phaeophyscia kairamoi, ph. hispidula and ph. cernohorskyi often noted in italian orchards are common and abundant in alpine regions, and in this situation impact of elevation on their notes in the orchards could play the role. despite records of some rare lichens for particular countries (e.g., bacidia rubella, melanelixia glabra, physconia perisidiosa and usnea hirta) most of identified species are widespread. such situation seems to characterized non-forested rural areas (giordani 2006), in which also orchards can be distinguished. species diversity dwindle to rather simplified flora due to also agricultural (loppi, de dominicis 1996). on the other hand records of some species included to the national/regional red lists confirm some observations about importance of anthropogenic habitats for occurrence of some valuable lichens (e.g., lush et al. 2005). lichenbiota of slovakia and poland is much close to each other in comparison with results obtained from italy. it can be explained by more similar climatic conditions of both countries connected with their geographical localization. on the other hand similarity of lichens records in slovak and italian orchards can be connected with altitudinal influence on the distribution of some species. both in slovakia and italy orchards were situated mainly or only in mountains regions where specific conditions could favorable some common species for both countries. most of recently noted lichens were observed during earlier studies conducted on epiphytes of fruit trees (tab. 2). mainly ubiquitous species with a broad geographical range were found in most of the investigated orchards: candelariella xanthostigma, phaeophyscia orbicularis, ph. nigricans, physcia adscendens, ph. tenella, xanthoria parietina, and x. polycarpa. all of above mentioned taxa are mesophytic or xerophytic and grow in rather sun-exposed sites (nimis, martellos 2008). considering previous data the species recorded mainly at apple orchards are: caloplaca cerina, lecania cyrtella, and xanthoria candelaria. especially caloplaca epiphytic lichens of apple orchards 157 table 2 bibliography of lichens recorded during the present study on malus sp., both solitary and in orchards, in poland, slovakia, and italy species bark of malus sp. solitary bark of malus sp. in orchard acrocordia gemmata kiszka, piórecki 1991, 1994; suppan, mayrhofer 2002 kościelniak 2004 amandinea punctata rydzak 1970; toborowicz 1976; lipnicki 1982; czyżewska 1998; czarnota 2000; suppan, mayrhofer 2002; guttová 2005; john 2006; szymczyk, zalewska 2008 bartók 1999; kiszka 1999; kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009 arthonia radiata mészáros et al. 1984; bartók 1999; lush et al. 2005 bacidia rubella kiszka, piórecki 1991; śliwa 1998; czarnota 2000; suppan, mayrhofer 2002; guttová 2005 suza 1936a; kuziel 1964; bartók 1999; kiszka 1999; kościelniak 2004; lush et al. 2005 caloplaca cerina rydzak 1970; suppan, mayrhofer 2002 suza 1936a; kuziel 1964; mészáros et al. 1984; bartók 1999 caloplaca citrina kuziel 1964 caloplaca holocarpa bartók 1999; lipnicki, sobieralska 2009 candelaria concolor rydzak 1970; toborowicz 1976; kiszka, piórecki 1991, 1994; śliwa 1998; czarnota 2000 kuziel 1964; mészáros et al. 1984; bartók 1999; kiszka 1999; kościelniak 2004; lush et al. 2005 candelariella reflexa śliwa 1998; lisická 1999; suppan, mayrhofer 2002; guttová 2005, szymczyk, zalewska 2008 kiszka, piórecki 1991; czarnota 2000; lush et al. 2005 candelariella xanthostigma rydzak 1970; lackovičová 1978; kiszka, piórecki 1991; lisická 1997; czarnota 2000; zalewska et al. 2004; guttová 2005; szymczyk, zalewska 2008 kuziel 1964; toborowicz 1976; czyżewska 1998; kiszka 1999; kościelniak 2004; pišút 2008 (unpubl.); lipnicki, sobieralska 2009 cladonia coniocraea suppan, mayrhofer 2002 kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009 cladonia fimbriata kiszka, piórecki 1991, 1994; suppan, mayrhofer 2002; szymczyk, zalewska 2008 kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009 dimerella pineti kiszka, piórecki 1991 kościelniak 2004 evernia prunastri endlicher 1830; zahlbruckner 1894; majeriková-hlaváčková 1957; rydzak 1970; toborowicz 1976; kiszka, piórecki 1991, 1994; suppan, mayrhofer 2002; szymczyk, zalewska 2008 kuziel 1964; lipnicki 2003; kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009 flavoparmelia caperata zahlbruckner 1894; rydzak 1970; kiszka, piórecki 1991, 1994; czarnota 2000; guttová 2005; john 2006 kuziel 1964; bartók 1999; kościelniak 2004; lush et al. 2005 hypocenomyce scalaris lipnicki 1982 kościelniak 2004; lipnicki, sobieralska 2009 hypogymnia physodes majeriková-hlaváčková 1957; halicz, kuziel 1958; rydzak 1970, toborowicz 1976; lackovičová 1978; lipnicki 1982; kiszka, piórecki 1991, 1994; czarnota 2000; suppan, mayrhofer 2002; zalewska et al. 2004; john 2006; szymczyk, zalewska 2008 kuziel 1964; bartók 1999; kiszka 1999; kościelniak 2004; lush et al. 2005; pišút 2008 (unpubl.); lipnicki, sobieralska 2009 hypogymnia tubulosa lisická 1999; czarnota 2000; suppan, mayrhofer 2002; john 2006 kuziel 1964; kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009 lecania cyrtella kuziel 1964; mészáros et al. 1984; czyżewska 1998; bartók 1999; kościelniak 2004 lecania naegelii suppan, mayrhofer 2002 kościelniak 2004; kuziel 1964; lush et al. 2005 158 d. zarabska et al. lecanora albella lush et al. 2005 lecanora carpinea rydzak 1970; toborowicz 1976; lackovičová 1978; kiszka, piórecki 1991, 1994; suppan, mayrhofer 2002; aptroot et al. 2006 halicz, kuziel 1958; kuziel 1964; mészáros et al. 1984; bartók 1999; kościelniak 2004; lipnicki, sobieralska 2009 lecanora chlarotera rydzak 1970; lisická 1997; suppan, mayrhofer 2002; aptroot et al. 2006; szymczyk, zalewska 2008 kuziel 1964; kościelniak 2004; lush et al. 2005 lecanora conizaeoides lackovičová 1978; lipnicki 1982 bartók 1999; kiszka 1999; kościelniak 2004; lipnicki, sobieralska 2009 lecanora expallens rydzak 1970; toborowicz 1976; lipnicki et al. 1991; kiszka, piórecki 1991, 1994; zalewska et al. 2004; john 2006 kiszka 1999; kościelniak 2004; lipnicki, sobieralska 2009 lecanora hagenii rydzak 1970; lisická 1997 kościelniak 2004 lecanora saligna śliwa 1998; czarnota 2000 czyżewska 1998; kościelniak 2004; lipnicki, sobieralska 2009 lecanora symmicta john 2006 bartók 1999; kościelniak 2004 lecidella elaeochroma rydzak 1970; lipnicki 1982; kiszka, piórecki 1991, 1994; suppan, mayrhofer 2002; guttová 2005; aptroot et al. 2006; szymczyk, zalewska 2008 kuziel 1964; bartók 1999; kiszka 1999; kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009 lepraria incana kiszka, piórecki 1991, 1994; zalewska et al. 2004, john 2006 bartók 1999; kiszka 1999; kościelniak 2004; lipnicki, sobieralska 2009 melanohalea elegantula suppan, mayrhofer 2002 kuziel 1964; kościelniak 2004 melanohalea exasperatula rydzak 1970; kiszka, piórecki 1991, 1994; czyżewska 1998; czarnota 2000; suppan, mayrhofer 2002; zalewska et al. 2004, john 2006; szymczyk, zalewska 2008 suza 1936a; kuziel 1964; toborowicz 1976; kościelniak 2004; lipnicki, sobieralska 2009 melanelixia fuliginosa majeriková-hlaváčková 1957; rydzak 1970; kiszka, piórecki 1991, 1994; czarnota 2000; suppan, mayrhofer 2002; zalewska et al. 2004; guttová 2005; john 2006; lisická et al. 2008 halicz, kuziel 1958; kuziel 1964; kościelniak 2004; lush et al. 2005 melanelixia glabra zahlbruckner 1894; lisická 1997; lisická et al. 2008 suza 1936a; kuziel 1964; kościelniak 2004 melanelixia subargentifera zahlbruckner 1894, 1905; kiszka, piórecki 1991; lisická 1997 kuziel 1964; kościelniak 2004 melanelixia subaurifera rydzak 1970; kiszka, piórecki 1991, 1994; suppan, mayrhofer 2002 kuziel 1964; kościelniak 2004; lush et al. 2005 opegrapha varia rydzak 1970 bartók 1999; lipnicki, sobieralska 2009 parmelia saxatilis majeriková-hlaváčková 1957; halicz, kuziel 1958; rydzak 1970; toborowicz 1976; lipnicki 1982; kiszka, piórecki 1991, 1994; pišút, lackovičová 1993; lisická 1997; czyżewska 1998; czarnota 2000; suppan, mayrhofer 2002; zalewska et al. 2004; john 2006; szymczyk, zalewska 2008 kuziel 1964; mészáros et al. 1984; kiszka 1999; bartók 1999; kościelniak 2004; lush et al. 2005; pišút 2008 (unpubl.) parmelina tiliacea zahlbruckner 1894; kiszka, piórecki 1991, 1994; aptroot et al. 2006; szymczyk, zalewska 2008 bartók 1999; kościelniak 2004 pertusaria albescens śliwa 1998; czarnota 2000; suppan, mayrhofer 2002; guttová 2005; john 2006 kościelniak 2004 pertusaria amara kiszka, piórecki 1991, 1994; suppan, mayrhofer 2002 mészáros et al. 1984; bartók 1999; kościelniak 2004; lush et al. 2005 phaophyscia chloantha kiszka, piórecki 1991 kościelniak 2004 table 2 cont. epiphytic lichens of apple orchards 159 phaeophyscia hirsuta kiszka, piórecki 1991, 1994; lisická 1999; lisická et al. 2008 kościelniak 2004 phaeophyscia nigricans rydzak 1970; toborowicz 1976; kiszka, piórecki 1991, 1994 kuziel 1964; kiszka 1999; lipnicki, sobieralska 2009 phaeophyscia orbicularis rydzak 1970; kiszka, piórecki 1991, 1994; suppan, mayrhofer 2002; guttová 2005; lisická et al. 2008; szymczyk, zalewska 2008 kuziel 1964; mészáros et al. 1984; bartók 1999; kiszka 1999; kościelniak 2004; lush et al. 2005; pišút 2008 (unpubl.); lipnicki, sobieralska 2009 phlyctis argena rydzak 1970; toborowicz 1976; lipnicki 1982; kiszka, piórecki 1991, 1994; czarnota 2000; suppan, mayrhofer 2002; zalewska et al. 2004; john 2006 kiszka 1999; kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009 physcia adscendens majeriková-hlaváčková 1957; rydzak 1970; toborowicz 1976; lackovičová 1978; lipnicki 1982; kiszka, piórecki 1991, 1994; czarnota 2000; john 2006; aptroot et al. 2006; szymczyk, zalewska 2008 mészáros et al. 1984; bartók 1999; kiszka 1999; kościelniak 2004; pišút 2008 (unpubl.); lipnicki, sobieralska 2009 physcia dubia rydzak 1970; toborowicz 1976, szymczyk, zalewska 2008 kuziel 1964; lipnicki 2003; kościelniak 2004; lipnicki, sobieralska 2009 physcia stellaris zahlbruckner 1894; rydzak 1970; lackovičová 1978; kiszka, piórecki 1991; lisická 1997; szymczyk, zalewska 2008 kuziel 1964; kiszka 1999; kościelniak 2004 physcia tenella rydzak 1970; toborowicz 1976; lipnicki 1982; kiszka, piórecki 1991, 1994; czarnota 2000; suppan, mayrhofer 2002; zalewska et al. 2004; john 2006; lisická et al. 2008; szymczyk, zalewska 2008 suza 1936a; kuziel 1964; czyżewska 1998; bartók 1999; kiszka 1999; kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009 physconia detersa majeriková-hlaváčková 1957; rydzak 1970; toborowicz 1976 kościelniak 2004 physconia distorta zahlbruckner 1894; kiszka, piórecki 1991, 1994; czarnota 2000; suppan, mayrhofer 2002; john 2006 kościelniak 2004; lipnicki, sobieralska 2009 physconia enteroxantha kiszka, piórecki 1991, 1994; czarnota 2000; john 2006, szymczyk, zalewska 2008 kościelniak 2004; lipnicki, sobieralska 2009 physconia grisea suza 1936b; rydzak 1970; toborowicz 1976; lisická 1997; john 2006; lisická et al. 2008 kuziel 1964; bartók 1999; kiszka 1999; kościelniak 2004; lipnicki, sobieralska 2009 physconia perisidiosa lisická 1997; czarnota 2000; john 2006 lipnicki 2003; kościelniak 2004; lipnicki, sobieralska 2009 pseudevernia furfuracea majeriková-hlaváčková 1957; toborowicz 1976; kiszka, piórecki 1991, 1994; lisická 1999; aptroot et al. 2006; john 2006 kuziel 1964; kościelniak 2004 scoliciosporum chlorococcum toborowicz 1976; kiszka, piórecki 1991, 1994; czarnota 2000; suppan, mayrhofer 2002 kiszka 1999; kościelniak 2004; lipnicki, sobieralska 2009 usnea hirta rydzak 1970; zahlbruckner 1894 kuziel 1964 xanthoria candelaria rydzak 1970; toborowicz 1976; śliwa 1998 kuziel 1964; mészáros et al. 1984; bartók 1999; kiszka 1999; kościelniak 2004; lipnicki, sobieralska 2009 xanthoria fallax rydzak 1970; kiszka, piórecki 1991, 1994; czarnota 2000 kuziel 1964; kiszka 1999; kościelniak 2004 xanthoria parietina szatala 1916; majeriková-hlaváčková 1957; rydzak 1970; lackovičová 1978; lipnicki 1982; kiszka, piórecki 1991, 1994; czyżewska 1998; lisická 1999; czarnota 2000; szymczyk, zalewska 2008 kuziel 1964; toborowicz 1976; bartók 1999; kiszka 1999; kościelniak 2004; lush et al. 2005; pišút 2008 (unpubl.); lipnicki, sobieralska 2009 xanthoria polycarpa rydzak 1970; kiszka, piórecki 1991,1994; aptroot et al. 2006; john 2006; szymczyk, zalewska 2008 kuziel 1964; toborowicz 1976; kościelniak 2004; lush et al. 2005; lipnicki, sobieralska 2009 table 2 cont. 160 d. zarabska et al. cerina should be paid attention; wirth (2002) treats this species as an indicator of orchards. notes about bacidina arnoldiana, cladonia merochlorophaea, micarea prasina, ochrolechia androgyna, phaeophyscia cernohorskyi, ph. hispidula, ph. kairamoi, physcia leptalea, strangospora pinicola, and xanthoria fulva are not given from malus sp. solitary and apple orchards in cited literature. among recorded species of special interest bacidia rubella should be distinguished. the species was recorded only once in one of the richest in lichens orchards in poland. mentioned above lichen was found associated with: candelariella reflexa, melanohalea exasperatula, parmelia sulcata, phaeophyscia nigricans, ph. orbicularis, and physcia adscendens. considering “red list of the lichens in poland” (cieśliński et al. 2006) bacidia rubella is included to the group of species under special attention. bark of apple trees seems to be one of the main substrate for bacidia rubella. the species was recorded only (suza 1936a; kuziel 1964; śliwa 1998) or mainly on this phorophyte (kiszka 1999; suppan, mayrhofer 2002; kościelniak 2004; guttová 2005). wirth (1995) considers apple bark in the orchards as an important habitats for its occurrence. among other species recorded in the polish orchards a record of strangospora pinicola is interesting. the species was observed only in one locality of all investigated orchards. previously no data about its occurrence in this kind of biotope were known. in poland strangospora pinicola is described with the status least concern (cieśliński et al. 2006). sun exposed sites on which species mainly occur (nimis, martellos 2008) can be found in the apple orchards (kuziel 1964) and probably it was main factor positively influenced on growing of strangospora pinicola in this ecosystem. in slovak orchards records of acrocordia gemmata, melanelixia glabra (vu) and usnea hirta (vu) should be highlighted. acrocordia gemmata occurs mostly in natural and semi-natural woods (e.g., lisická et al. 2008), the data from the past were scarse (pišút 1999). the second species occurs in the areas with higher precipitation, in open sites, mostly in semi-natural woods tolerated low degree of eutrophication (wirth 1995). although reported from relatively many localities in the malé karpaty mts. (lackovičová 1978) and also during mapping of epiphytic lichens in slovakia in 1970−1981 (pišút 1999), any further report is valuable. usnea hirta is one of the most frequently collected small species of the genus in slovakia after changes of air quality followed by transformation of industry after 1989. juvenile thalli can be found in urban areas occurring within nitrophilous xanthorion communities (e.g., nimis 1981; fos, clerc 2000) or in former areas influenced by industrial emissions. among commonly recorded lichens in italian orchards species from genus phaeophyscia should be distinguished. ph. hispidula and ph. kairamoi should be paid special attention. both of these taxa are restricted to the northern part of italy considering their distribution pattern in this country (nimis, martellos 2008). from italy ph. hispidula is reported as being at least very rare in particular regions of it’s occurrence (trentino-alto adige, lombardia). this species can be found on very acid to subneutral substrate in sites with very weak to weak eutrophication. notes of ph. kairamoi come from trentino-alto adige and friuli and it’s probably restricted to italian alps. it’s heliophilous species occurring in sun-exposed sites were recorded on base-rich bark (nimis, martellos 2008). epiphytic lichens of apple orchards 161 conclusions dissimilarity between number and recorded species in the studied sites can be the consequence of differences in genus and number of available phorophytes. in apple orchards the same bark and similar number of investigated trees (connected with restricted area of plots) minimizes this problem. it was the reason why so many common species were found between particular orchards and countries. orchards seems to create their own specific conditions which favors some groups of organisms, e.g., nitrophilous, heliophilous. nevertheless, occurrence of some rare species should pointed necessity of focusing more attention on this less studied type of agroecosystem. acknowledgements. dz acknowledges the ministry of education of the slovak republic, slovenská akaacknowledges the ministry of education of the slovak republic, slovenská akademická informačná agentúra (saia, apvv-51-040805, vega 0071), and ministry of foreign affairs in italy (3306 dell’11/09/2008) for financially support. authors are grateful dr. ivan pišút for use of his unpublished data from orchards, and anonymous reviewer for useful suggestions that helped improved the manuscript. references aptroot a., john v., wirth v. 2006. flechten und lichenicole pilze im dreiländereck ber der saarschleife mit neufunden aus lothringen, saarland und rheinland-pfalz (blam-exkursion 2005). herzogia 19: 63−76. bartók k. 1999. pesticide usage and epiphytic lichen diversity in romanian orchards. lichenologist 31 (1): 21−25. bielczyk u., lackovičová a., farkas e.e., lökös l., liška j., breuss o., kondratyuk s.ya. 2004. checklist of lichens of the western carpathians. biodiversity of the carpathians 1: 1−181. w. szafer institute of botany, polish academy of sciences, kraków. cieśliński s., czyżewska k., fabiszewska j. 2006. red list of the lichens in poland. 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(in:) a. zalewska, w. fałtynowicz (eds). lichens of the protected areas in the euregion niemen. “man and nature” association, suwałki: 5−50. porosty epifityczne sadów jabłoniowych w polsce, słowacji i włoszech streszczenie przeprowadzone badania porostów epifitycznych w trzydziestu sadach w polsce (10), słowacji (10) i włoszech (10) pozwoliły opracować listę 74 odnotowanych taksonów. najbardziej rozpowszechnione gatunki należą do grupy taksonów mezoi kserofitycznych oraz światłolubnych. w słowackich sadach zaobserwowano najwyższą liczbę gatunków oraz najwyższą liczbę gatunków wspólnych z przynajmniej jednym krajem. wydaje się, że kora jabłoni stwarza dogodne warunki dla rozwoju bacidia rubella, która razem ze strangospora pinicola były cennymi porostami odnotowanymi w polskich sadach. w sadach badanych na terenie słowacji szczególną uwagę zwróciły stanowiska acrocordia gemmata, melanelixia glabra i usnea hirta. phaeophyscia hispidula i ph. kairamoi to interesujące porosty zaobserwowane w sadach włoskich. 2014-01-01t11:49:10+0100 polish botanical society rhizocarpon lavatum and r. reductum (rhizocarpaceae, ascomycota), two misunderstood taxa found in the gorce mts (polish carpathians) anna matwiejuk department of botany, institute of biology, university of białystok świerkowa 20b, pl-15-950 białystok, matwiej@uwb.edu.pl matwiejuk a.: rhizocarpon lavatum and r. reductum (rhizocarpaceae, ascomycota), two misunderstood taxa found in the gorce mts (polish carpathians). acta mycol. 47 (1): 121–126, 2012. based on collections of p. czarnota deposited in the gpn herbarium, 8 species of rhizocarpon are reported from the gorce mountains. two of them, r. lavatum and r. reductum are reported from the gorce mts for the first time, instead of erroneously determined r. obscuratum. this species was previously misinterpreted due to taxonomic and nomenclatoric uncertainties. key words: rhizocarpaceae, lichens, taxonomy, western beskidy, national park indroduction new concepts of taxa and changes in nomenclature have triggered my studies on genus rhizocarpon. the present paper is the part of the taxonomic revision of representatives of the genus rhizocarpon in poland. the paper has been aimed to clarify the actual status of the species belonging to this genus, based on the latest innovations (fryday 2000; ihlen 2004). in particular, it is focuses on solving the problem of a group of species interpreted earlier as rhizocarpon obscuratum (ach.) massal. rhizocarpon obscuratum from the area of gorce mts was reported by glanc (1960), czarnota (2000), czarnota et al. (2005) and czarnota & wojnarowicz (2008). the thallus of rhizocarpon obscuratum was usually reported as lacking lichen substances (e.g., thomson 1997), although wirth (1995) mention that the thallus sometimes reacts k+yellow. dimensions of ascospores were reported as 14-53 × 6-21 μm (nowak, tobolewski 1975) or 20-32 × 9-15 μm (wirth 1995) and 22-50 × 9-18 μm (thomson 1997). according to fryday (2000) the specimens upon which lecidea petraea var. obscurata ach., the basionym of rhizocarpon obscuratum (ach.) massal., is based are acta mycologica vol. 47 (1): 121–126 2012 122 a. matwiejuk shown to be mostly referable to the species currently known as r. lavatum (fr.) hazsl. rhizocarpon reductum th. fr. is resurrected to accommodate specimens with small ascospores previously referred there. the name rhizocarpon obscuratum, which previously had an unclear relation to r. lavatum is formally excluded, because it proved to be synonym of fuscidea lygaea (ach.) v. virth & vězda (ihlen 2004). consequently, the name r. reductum (fryday 2000) has been used for the specimens of this complex producing stictic acid, and r. lavatum for the specimens lacking lichen substances. material and methods the study is based on the taxonomic revision of paweł czarnota’s collection made in the gorce mts in the years 1994-2005 and stored in the herbarium of the gorce national park in poręba wielka (gpn). the nomenclature of species is based on taxonomic studies by ihlen (2004). the species have been named according to santesson et al. (2004). the pigments arnoldiana-brown, atra-brown, atra-red (sensu meyer, printzen 2000) and macrocarpa-green (sensu fryday 2002) were recognized. chemical analyses were performed using a thin-layer chromatography (orange et al. 2001) with the use of solvents a and c. localities are given in the system atpol adapted for lichens (cieśliński, fałtynowicz 1993). microscopic measurements of anatomical features (ascospores length, breadth in μm and number of cells per ascospore) were made as characteristic of the diagnostic manual for rh. lavatum and rh. reductum. all ascospore measurements were made on mature ascospores and were also granted a definitively on the basis of the 24 spores measured for each species. all published specimens of the six species were previously correctly identified and their localities are in the following papers: glanc (1960), czarnota (2000), czarnota et al. (2005) and czarnota & wojnarowicz (2008). results and discussion as a result of the taxonomic revision, six species of rhizocarpon have been confirmed in the gorce mts, and two new, r. lavatum and r. reductum, have been distinguished based on their morphology and chemistry. confirmed species are: rhizocarpon distinctum th. fr. (glanc 1960; czarnota 2000; czarnota et al. 2005; czarnota, wojnarowicz 2008) rhizocarpon geminatum körb. (czarnota 2000) rhizocarpon geographicum (l.) dc. (czarnota 2000; czarnota et al. 2005; czarnota, wojnarowicz 2008) rhizocarpon lecanorinum anders (czarnota et al. 2005) rhizocarpon lavatum and r. reductum 123 rhizocarpon petraeum (wulfen) a. massal. (czarnota et al. 2005; czarnota, wojnarowicz 2008) rhizocarpon polycarpum (hepp) th. fr. (czarnota 2000; czarnota et al. 2005; czarnota, wojnarowicz 2008) rhizocarpon lavatum (fr.) hazsl., magyar. birod. zuzmó-flór.: 206 (1884) syn. rhizocarpon obscuratum f. lavatum (fr.) th. fr.; rh. orphninum (vain.) vain. (see ihlen 2004; santesson et al. 2004) this species is characterized by the areolate, grey, brown to rust-brown thallus, the flat to convex, rounded areoles, apothecia irregularly arranged, 0.5-1.0 mm diam., with mostly flat disc and distinct margin, epihymenium with intermixed atrabrown and macrocarpa-green pigments, exciple in section atra-brown pigmented, hymenium hyaline, 100-200 μm high, hypothecium dark brown (arnoldiana-brown), ascospores hyaline, muriform, with (8)10-14(-18) cells in optical view, 30.0-42.0 × 14-18 μm. spot test reactions: thallus k–, medulla i–. tlc: lichen products not detected. to date r. lavatum has been reported from poland only from beskid żywiecki mts: grupa wielkiej raczy (nowak 1998; bielczyk et al. 2004), tatra mts (flakus 2004, 2007) and the sudetes: karkonosze mts, mały śnieżny kocioł cirque (kossowska 2009a). revised specimens from morphological, anatomical and chemical features matches the description of r. lavatum by ihlen (2004). specimens examined (all previously recognized as r. obscuratum (ach.) massal.); on siliceous sandstones): western beskidy mts, gorce mts: ge 10 – the road close to olszowy stream, 690 m a.s.l., 1994, p. czarnota & j. kiszka (gpn/1894/94); the road on kosarzyska glade, n slope of kudłoń mt., in the catchment rosochy stream, 22 oct. 1997, p. czarnota (gpn/1925/94); ge 11 – hucisko glade, valley of kanina stream, 800 m a.s.l., 3 oct. 1994, p. czarnota (gpn/1372/94); the forest road close to satanowa polana glade, e slope of turbaczyk mt., 900 m a.s.l. and 940 m a.s.l., 2 july 1996, p. czarnota (gpn/1377/94 & gpn/1465/94); ge 20 – nature reserve turbacz, valley of olszowy stream, 810 m a.s.l., 21 june 1996, p. czarnota (gpn/1467/94); ge 21 – jaworzyna stream, in the catchment of łopuszanka stream, 860 m a.s.l., 5 may 1995, p. czarnota (gpn/1493/94); the road close to łopuszanka stream, near żubrowisko glade, 860 m a.s.l., 5 may 1995, p. czarnota (gpn 1767/94); ge 22 – s slope of twarogi, above settlement ochotnica dolna – barbarówka, 530 m a.s.l., 5 nov. 1999, p. czarnota (gpn/2153); lubań range, top of the lubań mt., 1200 m a.s.l., 23 june 2004, wojnarowicz 34. rhizocarpon reductum th. fr., lichenogr. scand. 1: 633 (1874) syn. rhizocarpon obscuratum auct. not. (ach.) a. massal. this species is characterized by the areolate, brown to greyish brown thallus, rounded flat to slightly convex areoles, apothecia irregularly arranged, 0.4-0.6 mm diam., with mostly flat, rough disc and distinct margin, epihymenium with intermixed atra-brown and macrocarpa-green pigments, exciple pigmented in section atrabrown and sometimes also macrocarpa-green, hymenium hyaline, 100-180 μm high, hypothecium dark brown (arnoldiana-brown), ascospores hyaline, muriform, with (6-)8-10(-12) cells in optical view, 24.0-26.0 × 8.0-10.0 μm. spot test reactions: thallus k+ yellow, c−; medulla i−, c−, pd+ orange. substances detected by tlc: stictic acid in thallus and apothecia. rhizocarpon reductum has been reported from poland many times, e.g. from the tuchola forest (fałtynowicz 1980), pojezierze choszczeńskie lakeland (lipnicki, tobolewski 1991), suwalski landscape park (kukwa, fałtynowicz 2002; 124 a. matwiejuk zalewska et al. 2004), białowieża forest (sparrius 2003), bieszczady national park (kościelniak, kiszka 2006), podlasie (matwiejuk 2007, 2009), karkonosze mts: czarny grzbiet ridge (kossowska 2009b) and beskid żywiecki (matwiejuk 2011). revised specimens from morphological, anatomical and chemical features matches the description of r. reductum by fryday (2000) and ihlen (2004). specimens examined (all previously recognized as r. obscuratum (ach.) massal.; on siliceous sandstones): western beskidy mts, gorce mts: ge 21 – nowa glade, below kiczora mt., 1220 m a.s.l., 6 nov. 1997, p. czarnota (gpn/1913/94); ge 22 – s slope of twarogi mt., above settlement ochotnica dolna – barbarówka, 530 m a.s.l., 5 nov. 1999, p. czarnota (gpn2131). taxonomic and nomenclatural remarks. rhizocarpon reductum sometimes resembles some forms of r. lavatum with small apothecia, but the presence of stictic acid usually makes it distinct. a comparison of the main features distinguishing rhizocarpon lavatum and r. reductum is presented in the table 1. the distinction of the two species – rhizocarpon lavatum and r. reductum is related to the changes in the taxonomy of rhizocarpon obscuratum auct. non (ach.) massal. complex, with hyaline and muriform ascospores (fryday 2000; ihlen 2004). rhizocarpon lavatum is similar to r. reductum, a name resurrected for the species previously commonly referred to r. obscuratum (ach.) a. massal. (fryday 2000). basionym for r. obscuratum, lecidea petraea var. obscurata ach. has been show to be based on a mixed type (fryday 2000) and to be regarded as a synonym of fuscidea lygaea (ach.) v. virth & vězda (ihlen, fryday 2002; ihlen 2004). an examination of the specimens upon which rhizocarpon obscuratum is based revealed that they are mostly referable to the species currently knows as r. lavatum. rhizocarpon reductum is resurrected for specimens formely placed under ‘r. obscuratum’ with small ascospores and the thallus containing stictic acid, whereas specimens with larger ascospores and a thallus lacking lichen substances are more referred to r. lavatum (fryday 2000). table 1 comparison of the main features separating rhizocarpon lavatum and r. reductum rhizocarpon lavatum rhizocarpon reductum principal habitat siliceous sandstones siliceous sandstones thallus grey, brown to rust-brown, areolate brown to greyish brown, areolate chemistry: lichen products not detected stictic acid spot test reactions thallus k– thallus k+ yellow diameter of apothecia (mm) 0.5-1.0 0.4-0.6 ascospores: lenght 30.0-42.0 24.0-26.0 breadth 14.0-18.0 8.0-10.0 number of cells in optical sections (8-)10-14(-18) (6-)8-10(-12) location of the insoluble lichen pigments hypothecium arnoldiana-brown arnoldiana-brown epihymenium atra-brown and macrocarpagreen atra-brown and macrocarpagreen exciple atra-brown atra-brown rhizocarpon lavatum and r. reductum 125 acknowledgements. i would like to express my warmest thanks to paweł czarnota, for facilities of collection of rhizocarpon. i wish to express my thanks to the reviewer for his precious remarks and advice. references bielczyk u., lackovičová a., farkas e.e., lőkös l., breuss o., kondratyuk s. ya. 2004. checklist of lichens of the western carpathians. w. szafer institute of botany, polish academy of sciences, kraków. cieśliński s., fałtynowicz w. 1993. note from editors.(in:) s. cieśliński, w. fałtynowicz (eds). atlas of the geographical 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2000. proposal for a standarized nomenclature and characterization of insoluble lichen pigments. lichenologist 32: 571–583. nowak j. 1998. porosty beskidów wyspowego i żywieckiego, pasma jałowca i masywu babiej góry. monogr. bot. 83: 1–131. nowak j., tobolewski z. 1975. porosty polskie. opisy i klucze do oznaczania porostów w polsce dotych-porosty polskie. opisy i klucze do oznaczania porostów w polsce dotychczas stwierdzonych lub prawdopodobnych. pwn, warszawa-kraków. orange a., james p. w., white, f. j. 2001. microchemical methods for the identification of lichens. british lichen society, london. 126 a. matwiejuk santesson r., moberg r., nordin a., tønsberg t., vitikainen o. 2004. lichen-forming and lichenicolous fungi of fennoscandia. museum of evolution, upssala university. sparrius l. b. 2003. contribution to the lichen floras of the białowieża forest and the biebrza valley (eastern poland). herzogia 16: 155–160. thomson j. w. 1997. american arctic lichens 2. the microlichens. madison, university of wisconsin press, 675 pp. wirth v. 1995. die flechten baden-württembergs. bd.1-2, ii aufl. stuttgart, verl. eugen ulmer. zalewska a., fałtynowicz w., krzysztofiak a., krzysztofiak l., picińska-fałtynowicz j. 2004. lichens of suwalski landscape park. (in:) a. zalewska, w. fałtynowicz (eds). lichens of the protected areas in the euroregion niemen, “man and nature” association, suwałki: 5–50. rhizocarpon lavatum i r. reductum (rhizocarpaceae, ascomycota), dwa nieznane dotąd taksony stwierdzone w gorcach (polskie karpaty) streszczenie w zbiorach zielnikowych gpn z gorców stwierdzono 8 gatunków rhizocarpon (r. distinctum, r. geminatum, r. geographicum, r. lavatum, r. lecanorinum, r. petraeum, r. polycarpum, r. reductum). po raz pierwszy dla tego terenu podano dwa gatunki rhizocarpon lavatum i r. reductum. wyróżnienie nowych gatunków związane jest ze zmianą ujęcia taksonomicznego i nomenklatorycznego reprezentantów grupy rhizocarpon obscuratum auct., charakteryzujących się bezbarwnymi i murkowatymi zarodnikami (fryday 2000; ihlen 2004). w oznaczeniu tych gatunków wykorzystano chromatografię cienkowarstwową (tlc), która pozwoliła określić ich cechy chemiczne: obecność kwasu stiktowego u r. reductum i brak wtórnych metabolitów u r. lavatum. pierwszy z tych gatunków charakteryzuje się ponadto mniejszymi i mniej podzielonymi zarodnikami [24.0-26.0×8.0-10.0 μm, (6-)8-10(-12) komórek], podczas gdy zarodniki r. lavatum osiągają rozmiary 30.0-42.0×14.0-18.0 μm i są podzielone na (8-)1014(-18) komórek. 2014-01-02t12:08:22+0100 polish botanical society analysis of indole compounds in fruiting bodies and in mycelia from in vitro cultures of calocera viscosa (basidiomycota) bożena muszyńska and katarzyna sułkowska-ziaja chair and department of pharmaceutical botany, collegium medicum, jagiellonian university medyczna 9, pl-30-688 kraków, muchon@poczta.fm muszyńska b., sułkowska-ziaja k.: analysis of indole compounds in fruiting bodies and in mycelia from in vitro cultures of calocera viscosa (basidiomycota). acta mycol. 47 (1): 57–64, 2012. calocera viscosa (pers.: fr.) fr. (basidiomycota) from dacrymycetaceae family is a widespread species of mushroom in poland. the aim of this study was to investigate the content of indole compounds in fruiting bodies and in mycelium cultured in vitro on solid and liquid medium of this species. fruiting bodies of calocera viscosa were collected in coniferous forests in south poland and were used to derive in vitro cultures. the optimal medium composition for cultures was determined. fresh material: fruiting bodies and mycelium from culture in vitro was frozen and then dried by lyophilization. the crushed dry biomass was extracted with petroleum ether to remove oil fraction, material was dried and extracted with methanol. analysis of indole compounds was performed in methanol extracts using chromatographic methods: tlc, uv vis, eims and hplc. this analysis presented in all three extracts the following indole compounds: l-tryptophan, 5-hydroxytryptophan, 5-methyltryptophan, melatonin and indole (contents fluctuated in the range: 0.37 to 11.88 mg/100 g d.w.). 5-hydroxytryptophan contents in all extracts were significant and amounted to 11.88 mg/100 g d.w. in fruiting bodies, and 11.42 in mycelium from liquid cultures and and 10.59 in mycelium from solid cultures. in addition, the fruiting bodies and mycelium from cultures on liquid medium revealed the presence of serotonin (0.39 and 3.19 mg/100 g d.w. respectively). key words: β,β-carotene, 5-hydroxytryptophan, mushroom, mycelial culture, serotonin introduction since the beginning of human evolution fruiting bodies of mushrooms belonging to the taxon basidiomycota are appreciated for flavor and texture but especially for their chemical and dietary properties. mushrooms are a rich source of a variety of biologically active compounds belonging to both primary and secondary metabolites acta mycologica vol. 47 (1): 57–64 2012 58 b. muszyńska and k. sułkowska-ziaja (yang et al. 2001; wasser 2002; muszyńska et al. 2010). more than 140,000 species of mushrooms exist in natural sites, but less than twenty five species are widely accepted as food (barros et al. 2007). the mushroom glucans are well known for their immunomodulatory properties and are used for anticancer therapy (sułkowska-ziaja et al. 2005; zaidman et al. 2005; muszyńska et al. 2011a). antitumor and cytostatic properties presented numerous terpenoids, especially sesquiterpenes and triterpenoids. a number of recent mycochemical papers have reported on the presence of many different terpene compounds (barros et al. 2008; liu 2005). the antioxidants found in edible mushrooms were flawonoids, phenolic compounds, ascorbic acid, tocopherols and carotenoids. recently, muszyńska (muszyńska et al. 2007; muszyńska et al. 2009; muszyńska et al. 2011b) have hinted at the occurrence of numerous non-hallucinogenic indole compounds in edible, conditionally edible and inedible basidiomycota species. calocera viscosa (pers.:fr.) fr. (yellow stagshorn) is a species common in polish, european and asiatic coniferous forests, which develops intensely yellow-orange branched, bush-like fruiting bodies. this species occurs from june to november on dead conifer wood in mixed and coniferous forests, mainly in mossy spruce and pine stumpsas saprotrophs. c. viscosa is a good source of free exoand endogenous amino acids, indole compounds, sterols (especially ergosterol) and unsaturated fatty acids (muszyńska 1999). although the carotenoids content in calocera viscosa fruiting bodies growing in poland were estimated spectrophotometrically at 2.46 μg/g f.w. by czeczuga (1980) at the end of the seventies years of the 20th century. barros (2008) demonstrated that of six basidiomycota species collected from natural sites in the northwestern portugal, cantharellus cibarius (chantarelle) contained the greatest amounts of β,β-carotene (13.56 μg/g d.w.) while the contents of this metabolite in the remaining species ranged from 1.95 12.77 μg/g d.w. the contents of this very important antioxidant were analyzed in extracts from fruiting bodies of c. viscosa and in extracts from mycelium of in vitro cultures (muszyńska et al. 2012). these contents are higher than those determined by czeczuga (1980). the cultures were maintained under different conditions to optimize biomass growth and to establish the most beneficial conditions for β,β-carotene accumulation. β,β-carotene content in biomass from solid cultures was comparable with that found in fruiting bodies (7.1 and 7.5 μg/g d.w., respectively). mycelia from liquid cultures contained half of that β,β-carotene amount which equaled 3.5 μ/g d.w. polysaccharides extracted from the mycelial culture of c.viscosa and administrated intraperitoneally into mice at dosage of 300 mg/kg inhibited the growth of sarcoma 180 and erlich solid cancers by 90% (ohtsuka et al. 1973). the aim of the present study was to initiate calocera viscosa culture in vitro, to determine optimal conditions for mycelia growth and to evaluate the indole compounds contents in extracts of fruiting bodies and mycelia from calocera viscosa in vitro cultures. indole compounds have attracted much interest recently because in vitro and in vivo studies suggest that they have a variety biological properties, which play important functions in the maintenance of human health. these compounds are antioxidants, antidepressants, anticancer, being tissue hormones and neurotransmitters. serotonin is a long known compound playing the role of a regulator of sleep, body temperature, mood, maturation and regeneration and an inhibitor of cell aging, thereby contributing to general strengthening of the immune system. its daily dose when used, for instance, as an antidepressant drug ranges from 100 to 200 mg (mosovich analysis of indole 59 et al. 2008). recent reports have revealed further important biological aspects of serotonin action, including its usefulness in prevention of alzheimer’s disease and the antioxidant action (ouchi et al. 2009). indole compounds and their derivatives play also important role as analgesic and anti-inflammatory medicines. materials and methods origin of fruiting bodies. the studies were conducted on fruiting bodies of calocera viscosa (pers.: fr.) fr. collected at natural sites in mixed and coniferous forests in southern poland (brodła near kraków) (deposited in the department of pharmaceutical botany, jagiellonian university, collegium medicum, kraków, poland). initial culture. initial cultures were derived from explants originating from the top parts of branched, bushlike fruiting bodies of calocera viscosa. these pieces of fruiting bodies were sterilized with 70% ethyl alcohol and placed on petri dishes with solid oddoux medium (oddoux 1957). cultures were incubated at a temperature 25 +/2° c under 12-h light (900 lx)/12 dark cycle and were subcultured every second week. for more details see muszyńska (muszyńska et al. 2009). experimental in vitro cultures. solid culture was maintained on petri dishes, on oddoux medium (oddoux 1957). stationary liquid culture was established from the solid culture by transferring 0.1 g of mycelium into an erlenmayer flask (500 ml) containing 250 ml of liquid oddoux medium. both types of experimental cultures were maintained under the same conditions as initial culture and were subcultured every two weeks. extraction. lyophilized and powdered fruiting bodies and mycelia from calocera viscosa in vitro cultures on solid and liquid media were extracted in a percolator with petroleum ether to remove oil fraction according to the procedure developed in our laboratory (muszyńska et al. 2007). after this extraction, biomass was dried and extracted by methanol in percolator for 24 h. extracts were concentrated to 5 ml by distillation in a vacuum evaporator under reduced pressure. for the purification of the extracts we used tlc method on aluminum – baked silica 60 plates (merck, art. no 1.05554.0001), on which 1 ml of the extracts was loaded and chromatograms were developed in mobile faze: n-butanol / acetic acid / water (12 : 3: 5 v/v/v). spots were identified at λ=254 nm. obtained fractions were analyzed by uv-vis, eims and hplc method. uv vis analysis of indole compounds. purified by preparative tlc method extracts of fruiting bodies and mycelia from cultures in vitro on solid and liquid medium were analyzed for the presence of indole compounds by spectrophotometry using uv-vis apart: uv-vis cary varian spectrophotometer. absorption measurements were carried out in the λ = 200-500 nm; solvent: methanol ar; standards: indole compounds manufactured by sigma-aldrich. in all three extracts was found an increase of the growth of an absorption maximum (for example at λmax = 227 and λmax = 254) characteristic for indole compounds. eims analysis of indole compounds. electron impact mass spectrometry analysis (eims) was performed at the regional laboratory of physicochemical analyses and structural research. apparatus: high resolution mass spectrometer with 60 b. muszyńska and k. sułkowska-ziaja options: ei, esi, gc-m, finnigan mat 95s. in the results of these studies were obtained the spectra for the methanol extracts from the fruiting bodies and mycelia from cultures on solid and liquid medium. the eims spectra for the methanol extracts from the fruiting bodies and mycelia from in vitro cultures on solid and liquid medium containing peaks characteristic for indole compounds (m/e=115, m/e=129, m/e=130, m/e=135, m/e=143, m/e=157). estimation of indole compounds by the hplc method. contents of indole compounds in extracts from fruiting bodies and in mycelia maintained in in vitro cultures on solid and liquid medium were determined after preliminary separation with preparative tlc method. contents of l-tryptophan, 5-hydroxytryptophan, 5-methyltryptophan, serotonin, melatonin, tryptamine, kynurenic acid, kynurenine sulfate, indoleacetic acid, β-indoleacetonitrile, indole and kynurenine were determined according to the procedure developed by kysilka and wurst (1985) with our modifications (muszyńska et al. 2009). briefly, the analytical conditions were as follows: hplc apparatus: hitachi; pump: l-7100; column: purospher® rp-18 (4 x 200 mm, 5 μm) thermostated at 25°c. the solvent system used were: methanol/water/ammonium acetate (15:14:1 v/v/v); flow rate:1ml/min. detection was carried out in a uv detector, using λ=280 nm; standards: manufactured by sigma-aldrich. standards solutions were prepared in hplc grade methanol. the identification of the indole compounds was made by comparing the retention times of samples peaks with standards. the results are expressed in mg/100 g of dry weight, calculated by internal normalization of the of the chromatographic peak area. results we established that good mycelial mass growth of calocera viscosa could be obtained in agitating liquid cultures and solid cultures on modified oddoux (1957) medium at 25 +/2°c under 16 h photoperiod (900 lx/8 h dark). a 20-fold fresh biomass growth in cultures on solid medium and a 15-fold growth in liquid cultures were obtained within a 14-day growth cycle. the biomass growth in the initiated cultures averaged 8.3 g d.w./1 l of medium. the obtained biomass increments and dynamics of mycelium growth did not differ from the results that we obtained for sarcodon imbricatus l. (sułkowska-ziaja 2010), xerocomus badius (fr.) kühn. ex gilb., tricholoma equestre (l.: fr.) kumm. (muszyńska et al. 2009) and cantharellus cibarius fr. cultures studied earlier. the in vitro cultures of these species were used for evaluation of qualitative and quantitative composition of non-hallucinogenic indole compounds and proved to be a valuable model for investigation of their metabolism. the present study is an extension of the previous studies on accumulation of indole compounds and is the first report about their presence in: fruiting bodies and in mycelia of calocera viscosa cultured in vitro. the identity of indole compounds was confirmed on the basis of their parameters tlc, uv-vis, eims and hplc methods. the hplc method was used for quantitation of indole compounds and optimum conditions were established by this method for separation: l-tryptophan, 5-hydroxytryptophan, 5-methyltryptophan, analysis of indole 61 serotonin, melatonin, tryptamine, kynurenic acid, kynurenine sulfate, indoleacetic acid, β-indoleacetonitrile, indole and kynurenine in extracts from fruiting bodies and in mycelia from in vitro cultures on solid and liquid medium. this analysis presented in all three extracts the following five indole compounds: l-tryptophan, 5-hydroxytryptophan, 5-methyltryptophan, melatonin and indole. in addition, the fruiting bodies and mycelium from cultures on liquid medium revealed the presence of serotonin (0.39 and 3.19 mg/100 g d.w. respectively). results are presented in table 1. contents of indole compounds in fruiting bodies ranged from 0.39 to 11.88 mg/100 g d.w., in mycelium from liquid cultures from 0.37 to 11.42 mg/100 g d.w., and in mycelium from solid cultures from 0.39 to 10.59 mg/100 g d.w. 5-hydroxytrypophan contents in all extracts were significant and amounted 11.88 mg/100 g d.w. in fruiting bodies, and 11.42 in mycelium from liquid cultures and 10.59 in mycelium from solid cultures. the second metabolites because of their amounts was 5-methyltryptophan and presented: 3.39 mg/100 g d.w. in fruiting bodies, 3.34 in mycelium from solid medium and 3.36 in mycelium from liquid medium. the contents of ltryptophan and indole were very comparable and ranged from 1.19 mg/100 g d.w. to 1.79. melatonin was found in comparable but small amounts in all three extracts: 0.47 mg/100 g d.w. in fruiting bodies, 0.39 in mycelium from solid medium and 0.37 in mycelium from liquid medium. in none of the extracts: tryptamine, kynurenic acid, kynurenine sulfate, kynurenine, indoleacetic acid, β-indoleacetonitrile and tryptamine were found. discussion among indole compounds, indole alkaloids and hallucinogenic compounds originating from mushrooms have been the main focus of interest. studies of nonhallucinogenic indole compounds have concentrated on tryptophan, among other compounds. tryptophan was identified in many basidiomycota species (kohlmünzer et al. 2000). tryptamine, serotonin, 5-hydroxytryptophan, indoleacetic acid, β –indoleacetonitrile, melatonin, kynurenic acid and kynurenine sulfate have also been quite frequently identified (muszyńska et al. 2009). table 1 contents of indole compounds (mg/100 g d.w.) in fruiting bodies of calocera viscosa and in mycelia from cultures in vitro fruiting bodies of c.viscosa mycelium of c.viscosa from solid medium mycelium of c. viscosa from liquid medium inndole compounds mg/ 100 g d.w. melatonin 0.47 ± 0.02 0.39 ± 0.01 0.37 ± 0.01 l-tryptophan 1.26 ± 0.04 1.19 ± 0.01 1.79 ± 0.05 5-hydroxytryptophan 11.88 ± 0.19 10.59 ± 0.11 11.42 ± 0.20 5-methyltryptophan 3.39 ± 0.02 3.34 ± 0.03 3.36 ± 0.02 serotonin 0.39 ± 0.01 – a 3.19 ± 0.07 indole 1.26 ± 0.04 1.21 ± 0.02 1.25 ± 0.03 data presented as mean of three series ± se; a content lower than 0.001 mg/100 g d. w. 62 b. muszyńska and k. sułkowska-ziaja our earlier studies analyzed nonhallucynogenic indole compounds in fruiting bodies of the following edible species from natural habitats: boletus edulis bull.: fr , cantharellus cibarius fr., lactarius deliciosus (l.: fr.) gray, leccinum rufum (schaeff.) kreisel, suillus luteus (l.: fr.) roussel, xerocomus badius (fr.:fr.) kühner ex gillbert, and of commercial origin: agaricus bisporus (j.e. lange) imbach, pleurotus ostreatus (jacq.: fr.) p. kumm , and the species considered as conditionally edible: armillaria mellea (vahl:) p. kumm. ss. lato., lactarius deterrimus gröger and tricholoma equestre (l.: fr.) p. kumm. ss. lato. the contents of indole compounds in all these species were diverse and had a very wide variability range from 0.01 to 39.20 mg/100 g d. w. similar to that of c. viscosa (muszyńska et al. 2007; muszyńska et al. 2009; muszyńska et al. 2011a, b). mycelial cultures of three species: cantharellus cibarius, tricholoma equestre, xerocomus badius were established in our laboratory and the contents of compounds under study range from 0.01 to 20.49 mg/100 g d. w. found melatonin content in the extracts from fruiting bodies of calocera viscosa was comparable to the content of these compounds in the previously studied species (ranged from 0.08 to 1.29 mg/100 g d.w.). in mycelia of cultures in vitro melatonin was established only in the extract from mycelium of t. equestre (0.60 mg/100 g d.w.) and was comparable with the contents of this compound in the mycelia of c. viscosa from solid and liquid medium (0.39 mg/100 g d.w. and 0.37, respectively). in the case of tryptophan, the highest content of this compound was found in the fruiting bodies of a. mellea (4.47 mg/100 g d. w.), while the other examined fruiting bodies and mycelia from cultures in vitro (x. badius 0.83 mg/100 g d.w. and t. equestre 1.04) content were identical for all extracts obtained from c. viscosa. the content of 5-hydroxytryptophan was previously the highest in the currently studied extracts from fruiting bodies and mycelium c. viscosa. in other species the highest content of this compound was in p. ostreatus (2.08 mg /100 g d.w.). in the present analysis for first time we established in extracts from fruiting bodies and mycelium from culture in vitro of c. viscosa 5-methyltryptophan and indole. compared with previously studied species the content of serotonin in the fruiting bodies of c. viscosa was low 0.39 mg/ 100 g d.w., while for example in the fruiting bodies of s. luteus, l. rufum and c. cibarius ranged from 29 to almost 38 mg/100 g d.w. however, in the mycelium of c. viscosa from cultures on liquid medium found content of serotonin was higher than had previously been detected in mycelium of t. equestre (3.19 mg/100 g d.w. and 0.59, respectively). tryptamine was labeled compound in most of the earlier fruiting bodies and mycelium of t. equestre and x. badius, but it was not detected in any extracts from c. viscosa. greater variety of indole compounds than in the fruiting bodies of c. viscosa was showed only in previously analyzed fruiting bodies of s. luteus, while mycelia from in vitro culture on solid and liquid medium of c. viscosa presented that amount of these metabolites was comparable with the amount indicated in the mycelia of t. equestre and x. badius (muszyńska et al. 2009). conclusions the obtained results indicate that calocera viscosa in vitro cultures can be a good model for the studies on accumulation and metabolism of indole compounds in mushrooms. these similar contents indicates also that it is possible to use in vitro analysis of indole 63 cultures as a model for studies on the physiological activity of above compounds. the comparable quantity of indole compounds obtained in the present study in c. viscosa mycelium with fruiting bodies collected from natural condition indicate that in vitro cultures are a good source of these compounds. high serotonin precursor 5-hydroxytrypophan contents in this material prove also a potential for the use of the mycelium cultured in vitro as a source of this physiologically important compound for humans. in vitro studies demonstrated that serotonin, melatonin and their indole derivatives (n-acetylserotonin, 6-methoxytryptamine) dose-dependently 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(ed.). 1957. recherches sur les mycéliums secondaires des homobasidiés en culture pure. imprimerie de trevoux, lyon. ohstuka s., ueno s., yoshikumi c., hirose f., ohmura y., wada t., fujii t., takahashi e. 1973. polysaccharides having an anticarcinogenic effect and a method of producing them from species of basidiomycetes. uk patent 1331513. ouchi y., yoshikawa e., futatsubachi m., yagi s., ueki t., nakamura k. 2009. altered brain serotonin transporter and associated glucose metabolism in alzheimere disease. j. nucl. med. 50: 1260–1266. sewerynek e , stuss m., oszczygieł k., kułak j , lewiński a. 2005. protective effect of indole compounds on lipopolysccharide induced lipid peroxidation in in vitro conditions. endokrynol. polska 4: 508–511. 64 b. muszyńska and k. sułkowska-ziaja sułkowska-ziaja k., muszyńska b., końska g. 2005. biologically active compounds of fungal origin displaying antitumor activity. acta polon. pharm. drug res. 6: 153–160. sułkowska-ziaja k. 2010. chemical analysis of sarcodon imbricatus (l.) p. karst fruiting bodies and mycelia from in vitro cultures and biological activity of their polysaccharide fractions. phd thesis. jagiellonian university collegium medicum, kraków, 97-107. wasser s.p. 2002. medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides. appl. microbiol. biotechnol. 60: 258–274. yang j-h., lin h-c., mau j-l. 2001. non-volatile taste components of several commercial mushrooms. food chem. 72: 465–471. zaidman b.z., yassin m., mahajana j., wasser s.p. 2005. medicinal mushroom modulators of molecular targets as cancer therapeutics. appl. microbiol. biotechnol. 67: 453–468. analiza zawartości związków indolowych w owocnikach i mycelium z kultur in vitro calocera viscosa (basidiomycota) streszczenie od tysięcy lat owocniki grzybów wyższych były wykorzystywane jako źródło pożywienia. obecnie mogą być pozyskiwane nie tylko ze stanu naturalnego i hodowli komercyjnych lecz również z kultur in vitro prowadzonych w odpowiednich warunkach. grzyby jadalne są coraz intensywniej badane ze względu na produkcję biologicznie aktywnych metabolitów wtórnych. biologicznie i leczniczo aktywne metabolity grzybów są używane w terapii tak poważnych schorzeń jak np.: choroby krążenia, cukrzyca, miażdżyca oraz choroby nowotworowe. niektóre z metabolitów wykazują działanie: przeciwwirusowe, przeciwbakteryjne, przeciwrobacze. w konwencjonalnej medycynie, w leczeniu onkologicznym najdłużej stosowane są polisacharydy grzybowe, stąd też są ich najlepiej poznanymi metabolitami. innymi grupami szerzej badanych związków są również liczne związki fenolowe, terpenowe, indolowe, witaminy, biopierwiastki (np. selen) o działaniu antyoksydacyjnym. obiektem badań w ramach niniejszej pracy był gatunek grzyba wielkoowocnikowego calocera viscosa (pers.: fr.) fr. (basidiomycota). owocniki tego gatunku występują pospolicie w lasach iglastych polski południowej. materiał do badań stanowiły owocniki zebrane w nowej białej (powiat nowy targ) oraz mycelium otrzymane z kultur in vitro, które wyprowadzono z owocników tego gatunku. kultury prowadzono na pożywce stałej i płynnej wytrząsanej wg oddoux w temperaturze 25 ± 2°c przez 14 dni w warunkach sztucznego oświetlenia o intensywności 900 lx. owocniki oraz zebrane biomasy z kultur in vitro wysuszono metodą liofilizacji, ekstrahowano eterem naftowym w celu usunięcia frakcji lipidowej, a następnie prowadzono ekstrakcję metanolem. uzyskane wyciągi metanolowe po zagęszczeniu, oczyszczono metodą preparatywnej chromatografii tlc na płytkach (dc alufolien kiesel gel f-254), a następnie analizowano na zawartość związków indolowych metodą hplc (identyfikacje związków indolowych dokonano metodami spektralnymi uv-vis i eims). na podstawie przeprowadzonych analiz stwierdzono obecność we wszystkich trzech ekstraktach następujących związków indolowych: tryptofanu, 5-hydroksytryptofanu, 5-metylotryptofanu, melatoniny i indolu (zawartości wahały się w przedziale: 0.37 do 11.88 mg/100g s.m.). związkiem indolowym występującym w największych ilościach we wszystkich ekstraktach był 5-hydroksytryptofan. dodatkowo w owocnikach i w mycelium z kultur na podłożu płynnym stwierdzono obecność serotoniny (odpowiednio: 0.39 i 3.19 mg/100g s.m). 2014-01-02t12:03:01+0100 polish botanical society some observations of slime moulds on wood and litter in beech forests dominika ślusarczyk department of mycology, university of łódź banacha 12/16, pl-90-237 łódź, dominika@biol.uni.lodz.pl ślusarczyk d.: some observations of slime moulds on wood and litter in beech forests. acta mycol. 45 (2): 239–246, 2010. the results of research into slime moulds in beech forest reserves in central poland are presented. thirty species of slime moulds directly associated with beech wood and beech litter were recorded. species associated with different decay phases of beech wood and litter were identified. key words: mycetozoa, ecology, beech forests, nature reserves introduction myxomycete species from central poland have so far been listed in two studies published by orzechowski (1966) and kalinowska-kucharska (1975). orzechowski gives 44 species from forest areas within the łódź town boundaries and some parks in łódź. kalinowska-kucharska provides a list of slime moulds collected by ławrynowicz in nature reserves in the łódź voivodeship, mostly in the tilio-carpinetum tracz. 1962 and potentillo albae-quercetum libb. 1933 associations (ławrynowicz 1973). present studies were conducted in nature reserves in central poalnd over four years. slime moulds occurring on fallen beech logs, trunks and stumps and beech litter were especially interesting. of them, six taxa are new to the area: fuligo leviderma, hemitrichia clavata, collaria arcyrionema (lamproderma arcyrionema), physarum leucophaeum, p. notabile, stemonitis pallida (seta, drozdowicz 2004). the aim of this study is to identify species of slime moulds associated with beech wood and beech litter during their development. acta mycologica vol. 45 (2): 239–246 2010 dedicated to professor barbara gumińska on the occasion of her eighty-fifth birthday 240 d. ślusarczyk study area the study area belongs to the wzniesienia łódzkie hills mesoregion according to kondracki (1998). investigations were conducted in three nature reserves and a natural site in central poland where european beech reaches the northern limit of its natural range (fig. 1). the wiączyń reserve was established in a 1 300 ha forest complex (las wiączyński forest) and covers an area of 8.40 ha. it comprises sections 170a, 170c and 176a, and is part of the former puszcza łódzka old-growth forest. the reserve protects a natural deciduous forest on the northern range limit of beech and fir, and comprises a shade forest. the field layer is composed of few species with a very small cover. this community type is the most representative of mixed beech forests and resembles acidic lowland beech forests (olaczek 1962; personal inf. 2004). the gałków reserve covers an area of 57.85 ha and comprises forest sections 240, 241 and 244a, b, c, d. it is one of the oldest reserves in the łódź voivodeship. it protects fagus sylvatica and abies alba which grow here at the limit of their geographic range. as reported by sowa and olaczek (1971), the primeval beechfir forest has been preserved in the reserve. however, it differs considerably from similar forests in southern poland which are distinguished by floral poverty, the absence of species associated with beech and the mountainous character. the field layer is formed by plants typical of beech forests, oak-hornbeam forests and other deciduous forests. approximately 29 beeches of natural and historic importance grow in the reserve. they are over 150 years old, with some as old as 300 years. the parowy janinowskie reserve is situated in the northern part of the janinów natural site complex, one of the largest sites of the natural occurrence of common beech (557 ha) in central poland (rutowicz, sowa 1976) and one of the areas of greatest value in the wzniesienia łódzkie landscape park (kurowski 1986, 1994, 1998). it comprises sections 37b, c, 38, 39a, b, f, 43b, 44a, b, and is part of a gentle n-facing slope covered with a network of original erosion ravines. luzulo pilosae-fagetum phytoceonoses cover considerable areas of the reserve (kurowski 1994). the paprotnia natural site covers 63.88 ha, comprises sections 221-224 and is situated on the line of the highest hills running from zgierz to the vicinity of brze-brzeziny. circaeo-alnetum, tilio-carpinetum and luzulo pilosae-fagetum forest associations were identified within the site. they cover the entire hilly area of the moraine plateau and its edge. a dense and healthy 90-year-old beech tree stand prevails. the edge, steep slopes and local elevations are covered by its poor variant; flat areas are covered by a variant with anemone nemorosa (łuczak, łuczak 2000a, b). the four areas are interesting sites of fagus sylvatica and have been studied by botanists, mycologists, mycosociologists and phytopathologists. herbarium collections of fungi gathered over a period of 50 years are deposited in the herbarium universitatis lodziensis (lod f). herbarium materials were analysed to identify life conditions of beech at the limit of its natural range. results will be published in successive papers. this study discusses myxomycetes and is the first in the series. 242 d. ślusarczyk current names of slime moulds were also used in the citations from source studies. the name under which a species was published by the original author is given in brackets. the specimens collected are deposited in the herbarium universitatis lodziensis (lod f). results list of species arcyria cinerea (bull.) pers. – wiączyń reserve: on fallen branches of fagus, vii 2002; parowy janinowskie reserve: on decaying beech wood, vii 2004. a. denudata (l.) wettst. –wiączyń reserve: on beech logs, ix 2000, v, xi 2001, v 2002, iii 2002, iii 2003; gałków reserve: on beech logs, x 2000, vii 2003; paprotnia natural site: on beech logs, x 2002. a. obvelata (oeder) onsberg – wiączyń reserve: on fagus wood; iv 2001, v, xi 2002, viii, x 2003; gałków reserve: on fagus wood, ix 2003; paprotnia natural site: on fagus wood, vii 2001. badhamia utricularis (bull.) berk. – wiączyń reserve: on decaying beech wood, x 2000. ceratiomyxa fruticulosa (o. f. müll.) t. macbr. – wiączyń reserve: on logs of fagus; ix 2000; gałków reserve: on logs of fagus, vii 2001, vii 2003; parowy janinowskie reserve: vii 2001; paprotnia natural site: vii 2001. collaria arcyrionema (rostaf.) nann.-bremek. ex lado – parowy janinowskie reserve: on a fagus stump, vii 2001. comatricha nigra (pers. ex j. f. gmel) j. schröt. – wiączyń reserve: on fallen branches of fagus, x 2003. diachea leucopodia (bull.) rostaf. – wiączyń reserve: on fallen beech leaves, viii-ix 2003. didymium melanospermum (pers.) t. macbr. – gałków reserve: on fallen beech leaves; viii 2003. fuligo leviderma h. neubert, nowotny & k. baumann – gałków reserve: on fallen beech leaves and fruits; viii 2001. f. septica (l.) f. h. wigg. – wiączyń reserve: on fallen beech leaves, ix 2000, viii 2001, ix 2003; gałków reserve: on fallen beech leaves, ii vii 2001, vii 2003; paprotnia natural site: on fallen beech leaves, vi-vii 2000, vi-vii 2001, vi 2002; parowy janinowskie reserve: on fallen beech leaves, viii 2001, vi 2002. hemitrichia clavata (pers.) rostaf. – wiączyń reserve: on a log of fagus; x 2000. lamproderma columbinum (pers.) rostaf. – paprotnia natural site: on decaying fagus wood, vii 2001. leocarpus fragilis (dicks.) rostaf. – wiączyń reserve: on pieces of fagus bark, viii 2001. lycogala epidendrum (l.) fr. – wiączyń reserve: on fagus trunks, vi, ix 2000, vii 2001; gałków reserve: on fagus trunks, x 2000, vii 2001, ix 2003; paprotnia natural site: on fagus trunks, viii 2000, vii, x 2001; parowy janinowskie reserve: on fagus trunks, viiviii 2001, x 2003. metatrichia vesparia (batsch) nann.-bremek. ex g. m. martin & alexop. – wiączyń reserve: on a decaying log of fagus, v 2001, iii-v 2002, xi 2002; gałków reserve: on a decaying log of fagus, iv 2001, ix 2003, ix 2004; paprotnia natural site: on fagus trunks, ii 2003; parowy janinowskie reserve: on fagus trunks, iii, vii-viii 2001. some observations of slime moulds 243 physarum album (bull.) chevall. – wiączyń reserve: on beech wood, viii 2003, ix 2004. p. cinereum (batsch) pers. – wiączyń reserve: on beech wood, x 2000, ix 2003. p. leucophaeum fr. – wiączyń reserve: on a beech log, ix 2000. p. notabile t. macbr. – wiączyń reserve: on a beech log, ix 2000. p. virescens ditmar in sturm – gałków reserve: on fallen beech leaves, viii 2001. reticularia lycoperdon bull. – wiączyń reserve: on decaying trunks of fagus, ix 2000, iv 2002, vi 2005. stemonitis axifera (bull.) t. macbr. – wiączyń reserve: on decaying fagus wood, x 2000, vii 2003, vi 2005. s. pallida wingate – parowy janinowskie reserve: on decaying fagus wood, viii 2001. stemonitopsis typhina (f. h. wigg.) nann.-bremek. – wiączyń reserve: decaying wood and fallen leaves of fagus, ix 2000, ix 2003, vi 2005; parowy janinowskie reserve: vii 2004. trichia decipiens (pers.) t. macbr.– wiączyń reserve: on beech wood, x 2003. t. favoginea (batsch) pers. – wiączyń reserve: on beech wood, iv 2001. t. scabra rostaf. – wiączyń reserve: on a decaying log of fagus, iii 2002, xi 2004. t. varia (pers. ex j. f. gmel.) pers. – wiączyń reserve: on beech wood, x 2000, x 2001; gałków reserve: on beech wood, iv 2001. tubulifera arachnoidea jacq.– wiączyń reserve: on a beech log, ix 2000, viii 2001; gałków reserve: on beech wood, vii 2001, vi 2002; parowy janinowskie reserve: viii 2001, vii 2004. discussion thirty species of slime moulds that occurred directly on beech trunks, stumps and logs and on beech litter composed of small beech twigs, bark pieces, fallen leaves and beech fruits were recorded. an ecological analysis of the results shows that some species, e.g., stemonitis axifera or arcyria denudata, preferred beech wood without bark in the optimal decay phase. other species, e.g., lycogala epidendrum, tubulifera arachnoidea and hemitrichia clavata, occurred on the bark of logs in the optimal and end decay phases. metatrichia vesparia and trichia scabra mostly colonised logs of fagus sylvatica in the end decay stage. the present results correspond to those reported in similar studies. in particular, investigations conducted by drozdowicz (1992a, b, 2008) confirm the observations that beech wood, and especially beech logs in the end decay phase, are a good substrate for the development of slime moulds. a comparison of the results collected in the ojców national park and central poland shows that 22 species occur in both areas on beech logs. only individual taxa distinguish the mountainous area from the lowland area. studies conducted by stojanowska and panek (2004a, b) in two nature reserves near wałbrzych (sw poland) also confirm that slime moulds develop very well on the wood of deciduous trees. thirty three species preferring beech wood and 31 species preferring limewood are listed. species that mostly occur on beech stumps include arcyria cinerea, a. denudata, a. incarnata, ceratiomyxa fruticulosa, comatricha pulchella, hemitrichia clavata, h. serpula, collaria 244 d. ślusarczyk arcyrionema (lamproderma arcyrionema), lycogala epidendrum, metatrichia floriformis, physarum album (p. nutans), p. leucophaeum, stemonitis fusca, stemonitopsis typhina, trichia sabra and t. varia. the occurrence of 29 species of slime moulds was recorded in studies conducted by miśkiewicz (2001) in the dentario glandulosae-fagetum and querco-fagetum communities in the bukowiec reserve in the carpathian mts. the taxa were associated with beech logs or lying stumps of fagus sylvatica: arcyria cinerea, a. denudata, hemitrichia clavata, lycogala epidendrum, metatrichia vesparia (m. vesparium) and stemonitis axifera. these species also occurred in the sites investigated by the present author and were recorded by stojanowska (1979), drozdowicz (1992b, 2008) and ing (1994), who reports 21 species associated with the wood of fagus sylvatica. stojanowska (1979) reports 42 myxomycete species on rotting beech wood from the opole region, góry kaczawskie mts, góry bialske mts, ślęża massif, wzgórza trzebnicko-ostrzeszowskie hills and karkonosze mts. however, these taxa have a broad range of occurrence. only seven of them colonise beech wood and do not occur on the wood of other deciduous trees. twenty five species developed in different combinations close together, both on small trunks and on large beech logs. stojanowska lists the following species in order of occurrence frequency: trichia varia, metatrichia vesparia (hemitrichia vesparium), h. clavata, stemonitopsis typhina (comatricha typhoides), trichia scabra, trichia denudata, stemonitis fusca, physarum album (p. nutans), lycogala epidendrum, trichia persimilis, t. favoginea, metatrichia floriformis (trichia floriformis), trichia botrytis, hemitrichia serpula, fuligo rufa, stemonitis axifera (stemonitis ferruginea), badhamia utricularis, lycogala conicum, trichia olivacea (a taxon currently considered to be a variant of t. decipiens), physarum leucopheum, physarum globuliferum, trichia decipiens, lycogala exiguum, arcyria affinis, ceratiomyxa fruticulosa. studies conducted in the białowieża old-growth forest (chlebicki et al. 1996) also showed that as many as 38 species of the 44 species recorded at the crypto programme research plot are associated with strongly decayed wood and mostly occur on wood previously colonised by fungi. the composition of slime moulds recorded in the litter is also characteristic. arcyria cinerea, a. denudata and comatricha nigra mostly occurred on beech twigs while diachea leucopodia, didymium melanospermum, fuligo septica and physarum virescens colonised beech leaves. stojanowska (1983) gives a list of species associated with the litter, also beech litter, in her long-term studies on slime moulds. these include species recoded in the study area: arcyria incarnata, comatricha nigra (on dry leaves and fallen twigs), diachea leucopodia, didymium melanospermum, leocarpus fragilis, physarum notabile (on beech bark lying on the forest floor) and physarum virescens. conclusions beech stumps, trunks, logs and litter are an excellent site for the development of many myxomycete species. the results confirm special preferences of slime moulds for beech wood observed by other authors. stemonitis axifera or arcyria denudata preferred bark-free beech wood in the optimal decay phase. other species, e.g., some observations of slime moulds 245 lycogala epidendrum, tubulifera arachnoidea, hemitrichia clavata, occurred on the bark of logs also in the optimal decay phase. metatrichia vesparia and trichia scabra most frequently colonised logs of fagus sylvatica in the end decay phase. diachea leucopodia, didymium melanospermum, fuligo septica, f. leviderma and physarum virescens mostly occurred in the beech litter. the investigations show that dead beech wood is a major and irreplaceable factor in forest diversity. it plays an important role in preserving particularly interesting and rare species of slime moulds that are also characteristic of wood and litter of fagus sylvatica. acknowledgements. the author thanks dr anna drozdowicz (kraków) for confirming the identification of slime moulds. i am very grateful to professor krystyna czyżewska (łódź) for helpful discussion and professor maria ławrynowicz (łódź) for valuable comments on the manuscript. this work was supported by the polish ministry of science and higher education (grant no. 2 p04g 08026). references chlebicki a., żarnowiec j., cieśliński s., 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gleba atro-fuliginea, inaequaliter marmorata, carnoso-friabili; ascis egloboso ovoideis, 4-6-sporis, raro 2-sporis; sporidis globosis, nigro-fuliginesis, 28-32 µm, diam. verruculis obtusis crebis compersis. hab. in insula cypro prope rudera templi aphroditis. odor gratus, sere sapae, h. e. musti cocti. after chatin (1897), a good number of works dealing with fungi were done in this island. last century, nattrass (1937) in a great work including plates, carried out the first list of cyprus fungi covering 164 species of anamorphic fungi, 37 species of ascomycetes and 145 species of basidiomycetes. as a continuation of that work, nattrass & papaioannou (1938) added 20 more species. the same year, dearness (1938), in a note, dealt with the species listed in the paper of nattrass (1937), but this was alredy known. altson (1956), in a research into acta mycologica vol. 48 (2): 207–218 2013 doi: 10.5586/am.2013.022 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 208 m. torrejón the cause of a root-rot illness on vicia faba, contributed with one species. the following year, 34 more species were studied in georghiou & papadopoullos (1957). sixteen years later, all the previous information was compiled in the work of zyngas (1973) in which, many species new to the island were added by the author. ten years later, willoughby (1983) dealt with rhizophlyctis rosea. in the new millennium, winey (2005) in an illustrated book about macromycetes describes a good bunch of species new to the island. in the next years, most authors dealt with single species originating from cyprus: tsopelas and nicolau (2005) reported heterobasidion annosum for the first time and tsopelas et al. (2008) reseached seiridium cardinale and neophytou & ioannou (2009) recorded graphiola phoenicis. nine species of agaricus were reported in the paper of momany et al. (2009). the same year, monany and gücel (2009) in an illustrated guide, showed information about 109 different species of macromycetes. also, edible and toxic fungi of cyprus, by loizides et al. (2011) is an important contribution improving the knowledge of macromycetes. finally, loizides (2008), loizides & kyriakou (2011) and loizides et al. (2012) delved into macromycetes species. materials and methods the materials were collected in 15 sites located mostly in central part of the island (tab. 1). descriptions of the main macroscopic characters were written down locally from fresh fungi specimens. all collections were dried and put into paper bags and preserved in the author’s herbarium (mth). table 1 location of the observation sites site no. site name utm location altitude (a.s.l.) date of collection 1 psilo dendra 36s 0488064 3861518 1171 m 17 nov. 2 troodos 36s 0488744 3863471 1698 m 17 nov. 3 armyrolivado 36s 0491050 3865271 1590 m 18 nov. 4 beginning of calydonian trail 36s 0488172 3863680 1583 m 18 nov. 5 trail to calydonian falls 36s 0487876 3662117 1324 m 19 nov. 6 road to platania 36s 0492829 3867191 1109 m 20 nov. 7 platania 36s 0493314 3866981 1074 m 20 nov. 8 foini 36s 0484965 3860776 953 m 21 nov. 9 kelefos bridge 36s 0476923 3860847 449 m 21 nov. 10 ayios nikolaos 36s 0476174 3858154 582 m 21 nov. 11 cedar valley 36s 0471562 3872069 1064 m 22 nov. 12 road to ayios mamas 36s 0500205 3856355 647 m 23 nov. 13 agia paraskevi 36s 0500918 3854126 597 m 23 nov. 14 asgata i 36s 0523631 3848544 158 m 24 nov. 15 asgata ii 36s 0521967 3849615 168 m 24 nov. in the laboratory, dried samples were rehydrated with the 5% sodium hydroxide solution. herbarium specimens have been studied in distilled water under carl zeiss jenaval light microscope (400-1000x) from the samples which were isolated under a kyowa tokyo stereo microscope (40-80x). meltzer’s reagent, congo red dye, 3% potassium hydroxide and lactophenol cotton blue solutions were used when necessary. fungi of cyprus 209 keys and monographs used for species identification, and references of the descriptions and illustrations of the species studied, have been indicated individually. species have been taxonomically arranged according to kirk et al. (2008) and latin names and authors’ epithets are given according to index fungorum. results and discussion twenty-five species of anamorphs, twenty-eight species of ascomycetes and twentysix species of basidomycetes have been arranged taxonomically. as the first group anamorphic fungi are listed. the most important contribution of the current report to the knowledge of cypriot fungi is the datailed information about the fourteen rare species: conferticium ochraceum, eutypa scabrosa, glonium lineare, hymenoscyphus calyculus, hypoderma incrustatum, hyphoderma nemorale, hysterographium mori, myriostoma coliforme, rectipilus cistophilus, stictis friabilis, stictis radiata, thanatephorus sterigmaticus, tomentella asperula and vuilleminia megalospora. anamorphic fungi alternaria sp. 1. site 14. on decaying bark of olea europaea. mth 1107. alternaria sp. 2. site 14. on decaying seeds of ferula communis. mth 1119. alternaria mouchaccae e. g. simmons (=ulocladium chlamydosporum) site 8. on wood from a fallen decaying branch of cistus creticus. mth 1098. boeremia hedericola (durieu & mont.) aveskamp, gruyter & verkley (=phoma hedericola) site 5. on living leaves of hedera helix. site 7. mth 1095. on alive leaves of hedera helix var. aureovariegata. mth 1552. cercospora smilacis thüm. site 5. on living leaves of smilax aspera. mth 1151. coleophoma oleae (d c.) petr. & syd. site 14. on decaying leaves of olea europaea. mth 1117. further information in llimona (1991). cylindrodendrum album bonord. site 15. on leaf litter of pistacia lentiscus. mth 1113. the branched shape of their conidiomata helps to separate the observated sample from cylindrocarpon, cylindrocladiella, gliocladiopsis and septomyrothecium. further information in sifert et al. (2011). dendrodochium citrinum grove site 3. on a dead branch of pinus nigra subsp. palliata. mth 1084. there is further information about this species in ellis & ellis (1997). embellisia aff. allii (campan.) e. g. simmons site 15. on pods of seeds of cistus creticus. mth 1112. the observated conidia are three-septate. 210 m. torrejón fusarium sp. site 7. on an alive leaf of quercus infectoria. mth 1092. helminthosporium microsorum d. sacc. site 1. on a decaying twig of quercus alnifolia. mth 1135. there is further information about this species in ellis & ellis (1997). microxiphium sp. site 14. on a decaying stem of ferula communis. mth 1111. monodictys putredinis (wallr.) s. hughes. site 13. on a decaying branch of quercus infectoria. mth 1158. on decaying twigs of quercus infectoria. mth 1159. you can get further information in seifert et al. (2011). passalora sp. site 14. on the underside of marcescent leaves of ceratonia siliqua. mth 1108. colonies hypophyllous from irregularly round to irregular, setose, dark brown up to 0.5 mm. in diameter. conida from straight to slighly curved, clavate or cylindricclavate, olivaceous-brown, smooth, 0-3 septate 7-20 x 3-4 µm. conidiophores geniculate 120-180 x 3-4 µm. in average, darker than conidia. phaeotheca fissurella sigler, tsuneda & j. w. carmich. site 7. on alive leaves of quercus alnifolia. mth 1089. further information in seifert et al. (2011). phaeosclera dematioides sigler, tsuneda & j. w. carmich. site 10. on a dead stem of asphodelus aestivus. mth 1105. site 14. on dead branches of prunus dulcis from the basal area close to the soil. mth 1109. to get more information about this species see de hoog et al. (2000) and seifert et al. (2011). polystigma fulvum pers. ex dc. site 5. on living leaves of prunus dulcis. mth 1150. further information in berhard et al. (1971 ) and a picture from this species in torrejón (2007), under the name of polystigma ochraceum. pseudoidium ceratoniae (comes) u. braun & r. t. a. cook site 14. on an alive fruit of ceratonia siliqua. mth 1116. further information in braun and cook (2012). sirodesmium olivaceum (link) tubaki site 3. on a decaying branch of cedrus brevifolia. mth 1083. you can see good plates from this species in ellis (2001) and seifert et al. (2011) under the name of coniosporium olivaceum. spilocaea oleaginea (castagne) s. huges site 14. on alive leaves of olea europaea. mth 1115. you can see a good plate and information of this species in dominguez (1957) under the name of cycloconium oleagineum. stigmina platani-racemosae (dearn.& barthol.) s. huges site 4. on a dead twig of platanus orientalis. mth 1087. there is further information about this species in ellis (2001). thermomyces lanuginosus tsikl. site 14. on litter from decaying fruits of ceratonia siliqua. mth 1114. the shape and ornamentation of its spores help to identify this species. there is further information in ellis (2001) and seifert et al. (2011). fungi of cyprus 211 torula herbarum (pers.) link site 8. on wood of cistus creticus. mth 1100. for further information see key, plate and description in ellis (2001). virgariella sp. site 13. on decaying leaves of quercus infectoria. mth 1120. ascomycota capnodiales sphaerulina sp. site 11. on the underside of decaying leaves of platanus orientalis. mth 1138. helotiales dermea cerasi (pers.) fr. site 11. small collection on decaying wood of arbutus andrachne. mth 1144. narrower ascospores and ascus help to separate this species from dermea prunastri. for further information see breitenbach & kränzlin (1984), ellis & ellis (1997) and medardi (2006). haglundia elegantior graddon site 1. on a decaying branch of quercus alinifolia. mth 1123. small collection. more information in ellis & ellis (1997). hymenoscyphus sp. site 11. only one ascocarp on a decaying twig of quercus alnifolia. mth 1142. hymenoscyphus calyculus (sowerby) w. phillips site 4. small collection on a dead branch of alnus orientalis. mth 1085. the spores of this species were described by breitenbach & kränzlin (1984) as guttulate, but in the revision of this work, dougoud (2000) according to baral & krieglsteiner (1985), the spores provided with globules of oil do not belong to this species. in the ascospores of the collection from cyprus, droplets were not observed. in all references under the name of hymenoscyphus conscriptus. hymenoscyphus fructigenus (bull.) gray site 5. single ascocarp on a decaying acorn of quercus alnifolia. mth 1145. for further information see breitenbach & kränzlin (1984). incrucipulum ciliare (schrad.) baral site 1. on the underside of dry leaves from a broken branch of quercus alinifolia. mth 1125. the shape of the spores helps to separate this species from lachnum fuscences, a similar specimen occurring in the same habitat. for further information see ellis & ellis (1997), under the name of dasyscyphus ciliaris. lachnum bicolor (bull.) p. karst. site 5. on decaying twigs of cistus creticus. mth 1145. mth 1155. there is further information in breitenbach & kränzlin (1984), under the name of dasyscyphus bicolor. mollisia amenticola (sacc.) rehm site 1. on a decaying catkin of alnus orientalis. mth 1127. there is further information in ellis & ellis (1997). 212 m. torrejón pyrenopeziza sp. site 5. on a decaying twig of cistus creticus. mth 1148. tapesia fusca (pers.) fuckel site 4. on a dead branch of alnus orientalis. mth 1085. there is more information in ellis & ellis (1997). hysteriales gloniopsis praelonga (schewein.) underw. & earle site 11. on decaying wood of arbutus andrachne. mth 1141. further information about this rare species can be found in checa (2004). glonium lineare (fr.) de not. site 8. on a dead branch of cistus creticus. mth 1103. for further information see plate and description in dennis (1981). hysterographium mori (schewein.) rehm site 1. small collection. on decaying wood of quercus alnifolia. mth 1130. further information about this rare species can be found in checa (2004). ostropales stictis friabilis (w. phillips & plowr.) sacc. & traverso site 13. on a dead twig of quercus infectoria. mth 1171. here is more information about this uncommon species in ellis & ellis (1997), under the name of schizoxylon friabilis. stictis radiata (l.) pers. site 11. sharing habitat with melittosporiella pulchella on decaying wood of arbutus andrachne. mth 1136. for further information see breitenbach & kränzlin (1984) and medardi (2006). pleosporales cucurbitaria spartii (nees ex fr.) ces. & de not. site 10. on dead stems of cistus creticus. mth 1101, mth 1102. further information can be found in checa (2004). leptosphaeria sp. site 13. on a decaying and very thin twig of quercus infectoria. mth 1164. trematosphaeria cisti naumov & dobrozr. site 8. on wood of cistus creticus. mth 1106. for further information see trotter & cash (1972). pleospora helvetica niessl site 3. on a dead branch of cedrus brevifolia. mth 1082. brown color of its ascospores helps to separate this species from plespora herbarum, a very similar one, but with yellowish or light brown ascospores. further information can be found in checa (2004). fungi of cyprus 213 rhitismatales coccomyces dentatus (j. c. schmidt & kunze) sacc. site 1. on the upper surface of decaying leaves of quercus alnifolia. mth 1126. there is more information in ellis & ellis (1997). colpoma quercinum (pers.) wallr. site 1. on the underside of decaying leaves of quercus alnifolia. mth 1129. it seems to be a great variability in the length of the spores of this species. in the present collection the shape of septate acospores is accoding to dennis (1981), but the length of them corresponds to the data given by breitenbach & kränzlin (1984). lophodermium pinastri (schrad.) chevall. site 7. on decaying needles of pinus brutia. mth 1093. black colour from dry apothecia and the size of spores helps to separate this species from other lophodermium which live in this habitat, such as l. pini-excelsae with grey apothecia and shorter spores, or l. seditiosum with pale grey apotecia when dry and longer spores, or l. conigenum with grey apotecia on the perimetral area and black when dry on the central one, and longer spores also. there is further information about rhytismataceae in minter (1986). melittosporiella pulchella höhn. site 11. on decaying wood of arbutus andrachne, sharing habitat with stictis radiata. mth 1136. on a decaying branch of arbutus andrachne. mth 1143. naemacyclus fimbriatus (schwein.) dicosmo, peredo & minter site 7. on scales of fallen decaying con of pinus brutia. mth 1089. site 13. mth 1161. more information in dennis (1981) under the name of lasiostictis fimbriata. naemacyclus minor butin site 5. on decaying needles of pinus brutia. mth 1154. more information in dennis (1981). propolis farinosa (pers.) fr. site 11. small collection on a decaying branch of arbutus andrachne. mth 1140. there is further information in breitenbach & kränzlin (1984), under the name of propolis versicolor. xylariales eutypa scabrosa (bull.) auersw. site 13. on a dead stem of cistus creticus. mth 1162. there is further information in breitenbach & kränzlin (1984). nemania confluens (tode) laessøe & spooner site 13. immersed in rotten wood of quercus infectoria. mth 1165. mth 1167. for furter information see ellis & ellis (1997), under the name of hypoxylon confluens. basidiomycota agaricales rectipilus cistophilus esteve-rav. & vila site 13. small but important collection. on a decaying twig of cistus creticus. mth 1163. external curly hairs help to identify this sample. in spite of this species is abundant in catalonia, where was described as a new species to the science, living 214 m. torrejón on decaying leaves and twigs of cistus monspeliensis and c. salvifolius, it might be the first time that it is collected outside catalonia. there is further information in vila et al. (1999). atheliales athelia decipiens (höhn. & litsch.) j. erikss. site 13. on rotten wood of quercus infectoria. mth 1166. there is further information in bernicchia & gorjón (2010). auriculariales exidiopsis calcea (pers.) k.wells site 5. on wood from a branch of pinus brutia. mth 1153. further information can be found in breitenbach & kränzlin (1986). cantharellales thanatephorus sterigmaticus (bourdot) p. h. b. talbot site 8. on a non decorticated dead stem of cistus creticus. mth 1096, mth 1104. its thick and long sterigmata is a great help to identify this rare species. further information can be found in bernicchia & gorjón (2010). corticiales lyomyces sambuci (pers.) p. karst. site 13. on a decaying branch of quercus infectoria. mth 1160. for further information see bernicchia & gorjón (2010). vuilleminia comedens (nees) maire site 1. on a decaying branch of quercus alnifolia. mth 1134. site 11. on a decaying branch of arbutus andrachne. mth 1139. further information can be found in bernicchia & gorjón (2010). vuilleminia macrospora (bres.) horstam (=corticium macrosporopsis jülich) site 8. on wood from branches of cistus creticus. mth 1096, mth 1099. it must be the most common species in cistus spp. as you can see in torrejón (2009). further information can be found in breitenbach & kränzlin (1986), as corticium macrosporopsis. vuilleminia megalospora bres. site 5. on a decaying branch of quercus alnifolia. mth 1149. further information about this species can be found in bernicchia & gorjón (2010). dacrymycetales dacrymyces variisporus mcnaab site 2. on wood from branches of pinus nigra subsp. palliata. mth 1131. clamp connections and bigger basidiospores help to separate this species from dacrymyces fungi of cyprus 215 stillatus, a similar one, which lives in the same habitat. further information can be found in breitenbach & kränzlin (1986). geastrales myriostoma coliforme (dicks.) corda site 11. two basidiomata, on soil very rich in humus beneath quercus alnifolia and platanus orientalis, close to cyclamen cyprius. mth 1137. it is always good news to find out this species, which is included as a candidate in the european council for the conservation of fungi’s red list of macomycetes. there is further information in calonge (1998) and sarasini (2005). polyporales hyphoderma incrustatum k. h. larss. site 2. on decaying wood from a log of pinus nigra subsp. palliata. mth 1133. the whitish to grweyish basidiomata of the studied species, grows on a dense forested area with high humidity level. habitat and colour of basidiomata help to separate this rare species from hyphoderma memorale a xerophytic one with whitish to cream basidiocarps. further information can be found in bernicchia (2010). hyphoderma nemorale k. h. larss. site 5. on decaying wood from a branch of quercus alnifolia. mth 1156. the whitish to cream ascocarp of the studied sample is thicker than the whitish to grweyish basidiomata of hyphoderma incrustatum. also, habitat and ecology are different. because the two species have been collected in this island, this fact was the most important help to separate one to another. there is further information in bernicchia & gorjón (2010). rigidoporus sanguinolentus (alb. & schwein.) donk site 2. on decaying wood of pinus nigra subsp. palliata. mth 1121, mth 1122. it is very easy to separate basidiocarps from substratum. on the other hand, its fusiform cystidia help to identify this species. further information can be found in bernicchia (2005) and ryvarden & gilbertson (1994). under the name of physisporinus sanguinolentus in the last one. pucciniales phragmidium mucronatum (pers.) schltdl. site 7. on alive leaves of rosa sp. mth 1088. the tapered shape of its apical papilla is an important taxonomical features of this species. you can see more information in cooke (1902) and grove (1913) under the name of phragmidium disciflorum. phragmidium violaceum (schultz) g. winter site 5. on alive leaves of rubus ulmifolius. mth 1147. there is further information in grove (1913) and minkevičius & ignatavičiūtė (1991). puccinia salviae unger site 13. on alive leaves of salvia sp. mth 1166. there is further information in gonzález-fragoso (1925) and minkevičius & ignatavičiūtė (1993). 216 m. torrejón puccinia sorghi schwein. (aecidium state) site 13. on an alive leaf of oxalis corniculata. mth 1173. there is further information in minkevičius & ignatavičiūtė (1993). russulales acanthophysium cf. minor (pilát) tellería site 2. on decaying wood of pinus nigra subsp. palliata. mth 1123. most of the basidiospores are in the average of the size of acnathophysium minor, but some of them are bigger and closer to the size of aleurodiscus cerussatus. further information can be found in bernicchia & gorjón (2010) and telleria & melo (1995). conferticium ochraceum (fr.) hallenb. site 2. on decaying wood of pinus nigra subsp. palliata. mth 1128. before this interesting collection, it was always collected on wood of picea abies. there is further information in bernicchia & gorjón (2010). peniophora cinerea (pers.) cooke site 14. on a decaying branch of ceratonia siliqua. mth 1110. further information can be found in bernicchia & gorjón (2010). peniophora lycii (pers.) höhn. & litsch. site 15. on a decaying branch of cistus creticus. mth 1118. three kinds of cystidia and larger basidiospores allow separate the studied sample from peniophora cinerea. further information can be found in bernicchia & gorjón (2010) and jülich (1989). peniophora quercina (pers.) cooke site 13. on a decaying branch of quercus infectoria. mth 1168. there is further information in bernicchia & gorjón (2010). peniophora meridionalis boidin site 13. on a decaying branch of quercus infectoria. mth 1170. the shape or its lamprocystidia is different from other species of this genus, of which live in the same habitat. you can get further information in bernicchia & gorjón (2010). stereum ochraceoflavum (schwein.) sacc. site 1. on wood from decaying branches of quercus alnifolia. mth 1132. for further information see breitenbach & kränzlin (1986). thelephorales pseudotomentella sp. site 13. on a rotten twig of cistus creticus. mth 1172. tomentella asperula (p. karst.) höhn. & litsch. site 5. on a decaying acorn of quercus alnifolia. mth 1157. you can get further information in kõljalg (1995). acknowledgments. the author would like to express his thanks to all the mycologist and colleagues from british mycological society and cyprus mycological association participants of the foray for unforgettable days. i also thank dr henry beker and dr stuart skeates for providing information about location of the sites. lastly, my deep thanks go to mr michael loizides, dr salih gücel and dr stephanos diamandis for providing cypriot bibliography. fungi of cyprus 217 references altson r. a. 1956. report on an investigation into the cause of a root-rot of broad beens (vicia faba) in cyprus. department of agriculture, the government of cyprus, nicosia, cyprus. baral h.o., krieglsteiner g.j. 1985. blausteine zu einer ascomyzeten-flora der bundesrepublik deutschland in söddeutschland gefundene inoperculate discomyzeten. beih. zeitschr. f. mykol 6: 1-226. bernhard r., grasselly c.h., leglise p., lansac m., marenaud c., 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warmia plain rafał szymczyk and anna zalewska department of botany and nature protection, university of warmia and mazury in olsztyn pl. łódzki 1, pl10-727 olsztyn, graphis22@poczta.onet.pl szymczyk r., zalewska a. lichens in the rural landscape of the warmia plain. acta mycol. 43 (2): 215–230, 2008. lichens and lichenicolous fungi in the rural landscape of the warmia plain were studied. lichen species were observed on old wooden fences, roadside trees, fruit trees, pylons, farm machinery, buildings and bridges. the analysed biota consists of 104 taxa with several noteworthy and rare lichens. key words: lichenized and lichenicolous fungi, anthropogenic habitats, n poland introduction the warmia plain is the most easterly mesoregion of the littoral gdańsk region. it is situated to the east and north-east of the elbląg high plain, and it occupies an area of around 640 m2 (kondracki 2001). the plain slopes down mostly to the north from the altitude of 60-70 m to 20 m above sea level and scarps down to the old prussian coast stretching along the vistula lagoon. the warmia plain is partially covered with varved clay from the recession period of the pomeranian phase of vistula glaciation, while the ground surface in the southern area near pasłęk consists of boulder clay. extensive farm fields and pastures separated by woods are the predominant features of the local landscape. the region is intersected by the rivers bauda and pasłęka whose deep valleys cut into the local relief. the lichen biota of nearly completely deforested rural areas is rarely individually studied (kuziel 1964). data concerning lichens in such habitats are found in most lichenological studies investigating large areas, but they are difficult to isolate from the general context (e.g. fałtynowicz and tobolewski 1989; cieśliński 2003; czyżewska 2003a). comparative data can be found in selected monographs which separately analyse the diversity of non-synanthropic and apophytic lichens (e.g. fałtynowicz 1992; bystrek and kolanko 2000) or in studies devoted to the synanthropization of lichen biota (articles from the collection of kiszka and piórecki 1994 (eds), e.g. kiszka 1994, fałtynowicz 1994; cieśliński and czyżewska 1998, olech 1998). acta mycologica vol. 43 (2): 215–230 2008 216 r. szymczyk and a. zalewska the warmia plain is weakly researched as regards its lichen population. the most abundant source of data is an unpublished study by woźniak (1983), which discusses mostly lichens on roadside trees along the main roads of the region. information on the localities of several species can also be found in the papers by sulma and fałtynowicz (1988), fałtynowicz and sulma (1994), as well as in atlases (tobolewski 1983, 1988). the objective of this study was to record species of lichens, lichenicolous and saprotrophic fungi growing on various anthropogenic substrates in a man-transformed rural landscape. study area the studied site was the village of nowica in the warmia plain in northern poland (fig. 1). this small village is situated in the wilczęta municipality, county of braniewo. it has a population of around 100 inhabitants and features housing typical of former german settlements. all farms and public utility buildings form a single cluster surrounded by farm fields and pastures. the village is intersected by a nameless river, a tributary of the bauda river. nowica is the crossing point of two provincial roads with old roadside oaks, awarded the status of natural monuments, as well as younger ash and linden trees. the investigated site is situated in square bd 08 of the atpol grid (modified for lichens by cieśliński and fałtynowicz 1993) inside an area mapped by lines with the following geographic coordinates: 54°10’53.9’’– 54°11’27.4’’n / 19°48’21.2’’–19°48’45’’e. fig. 1. location of the investigated area lichens in the rural landscape 217 materials and methods the experimental material was collected by the first author between 2001 and 2006 within the administrative boundaries of the village of nowica. during field observations, the author compiled records of the local biota and collected around 100 samples of lichen thalli from various substrates, including the bark of roadside trees, fruit trees, riverside trees, old wooden fences, farm buildings, field stakes, concrete fences, posts, bridges, roof tiles, asbestos-cement roofing material, granite border posts and metal parts of farm machinery. the collected material was analyzed by standard morphological and anatomical methods. the composition of secondary lichen metabolites was used to identify species of the genus lepraria and other lichens with sterile and crustose thalli. lichen substances were studied by thin-layer chromatography (orange et al. 2001). the applied nomenclature follows fałtynowicz (2003), excluding bacidia adastra (sparrius and aptroot 2003) and the following genera: coenogonium (kauff and büdel 2005), melanelixia and melanohalea (hawskworth et al. 2008) and piccolia (hafellner 2004). the collected material was deposited in the herbarium of the department of botany and nature protection, university of warmia and mazury in olsztyn (ols-l). results a total of 96 lichen species and 8 species of non-lichenized fungi (including 7 species of lichenicolous fungi and 1 species of saprotrophic fungi) were identified in the research area. below is an alphabetic list of the noted taxa. the number of records in the studied area and substrate types are given for every taxon. the relevant categories of threatened species, according to cieśliński et al. (2006), are indicated for vanishing lichens in poland. protected species (regulation of the ministry of the environment 2004) are marked with the letter “p”. lichenicolous fungi are marked with an asterisk (*), and saprobionts – with a plus sign (+). list of species acarospora fuscata (schrad.) th. fr. 2 records; on roof tiles; 12 july 2005 (ols-l 371). amandinea punctata (hoffm.) coppins & scheid. 22 records; on the bark of oak, apple tree, ash, horse-chestnut, linden, alder, poplar, willow and cherry tree, on wood and roof tiles; 12 july 2005 (ols-l). also recorded by woźniak (1983). anaptychia ciliaris (l.) körb. 3 records; on the bark of ash and oak; 14 july 2005 (field record, not collected), p. also recorded by woźniak ( 1983). anisomeridium polypori (m.b. ellis & everh.) m.e. barr 4 records; on the bark of ash, alder and willow; 14 july 2005 (ols-l 297). 218 r. szymczyk and a. zalewska *athelia arachnoidea (berk.) jülich 1 record; on thallus and apothecia of lecanora conizaeoides growing on an alder; 12 july 2005 (ols-l). bacidia adastra sparrius & aptroot 1 record; on the bark of apple tree; 25 dec. 2005, rev. m. kukwa (ols-l 416). bacidina phacodes (körb.) vězda 1 record; on the bark of willow; 14 aug. 2001 (ols-l 292). buelia griseovirens (turner & borrer ex sm.) almb. 4 records; on the bark of alder and wood; 20 aug. 2005 (ols-l 395). caloplaca citrina (hoffm.) th. fr. 3 records; on concrete; 18 aug. 2001 (ols-l 283). c. decipiens (arnold) blomb. & forssel 3 records; on cement-asbestos roofing material and concrete; 15 aug. 2005 (field record, not collected). c. flavocitrina (nyl.) a. e. wade 1 record; on concrete; 12 aug. 2005 (ols-l 398). c. holocarpa (hoffm.) a. e. wade 6 records; on the bark of elder, cement-asbestos roofing material, wood, concrete and metal; 12 july 2005 (ols-l). c. obscurella (j. lahm) th. fr. 1 record; on the bark of willow; nt; 15 aug. 2001 (ols-l 291). c. saxicola (hoffm.) nordin 7 records; on cement-asbestos roofing material and concrete; 1 march 2005 (ols-l 284). candelaria concolor (dicks.) stein 5 records; on the bark of linden, alder, willow, on wood; 1 march 2005 (ols-l). candelariella aurella (hoffm.) zahlbr. 5 records; on cement-asbestos roofing material, concrete and metal, 1 march 2005 (ols-l). c. reflexa (nyl.) lettau 20 records; on the bark of oak, apple tree, ash, maple, linden, alder, poplar, willow, on wood; 14 july 2005 (ols-l 296, 345). c. xanthostigma (ach.) lettau 12 records; on the bark of oak, apple tree, ash, maple, poplar, willow, on wood; 21 sept. 2005 (ols-l). c. vitellina (hoffm.) müll. arg. 3 records; on roof tiles and metal; 12 july, 21 sept. 2005 (ols-l 368, 373). also recorded by woźniak (1983). chaenotheca trichialis (ach.) th.fr. 1 record; on wood; 10 aug. 2005 (ols-l 385). nt category. +chaenothecopsis savonica (räsänen) tibell 1 record; on wood; 10 aug. 2005 (ols-l 386). lichens in the rural landscape 219 note. this species is usually parasitic or parasymbiontic on lichen thalli or on algae (e.g. sparrius 2003; groner 2006), but it can also grow on wood (see szymczyk 2007). cladonia fimbriata (l.) fr. 4 records; on apple tree bark and on wood; 25 dec. 2004 (ols-l 286). c. polydactyla (flörke) spreng. 1 record; on wood; 21 sept. 2005, (ols-l). coenogonium pineti (schrad. ex ach.) lücking & lumbsch 3 records; on the bark of alder; 14 july 2005, (ols-l). evernia prunastri (l.) ach. 11 records; on the bark of oak, pear tree, apple tree, ash, maple, linden and alder, on wood; 26 march 2005 (ols-l 332). nt category, p. also recorded by woźniak (1983). hypocenomyce scalaris (ach.) m. choisy 6 records; on the bark of oak, ash, alder, on wood; 12 july 2005 (field record, not collected). hypogymnia physodes (l.) nyl. 24 records; on the bark of elder, oak, pear tree, apple tree, ash, maple, linden, alder, willow, cherry tree, on wood; 20 july 2004 (field record, not collected). also recorded by woźniak (1983). h. tubulosa (schaer.) hav. 4 records; on the bark of oak and ash; 20 july 2004 (ols-l 300). nt category, p. lecania cyrtella (ach.) th. fr. 2 records; on the bark of elder; 20 sept. 2005 (ols-l 288). lecanora albescens (hoffm.) flörke 6 records; on concrete; 1 march 2005 (ols-l 347). l. allophana (ach.) nyl. 1 record; on ash bark; 20 july 2004 (ols-l 304). l. argentata (ach.) malme 6 records; on the bark of oak, maple and poplar, on wood, 7 aug. 2005 (ols-l 348). also recorded by woźniak (1983). l. carpinea (l.) vain. 11 records on the bark of elder, oak, ash, horse-chestnut, hazel, linden, poplar, cherry tree, on wood; 20 sept. 2005 (ols-l 346). also recorded by woźniak (1983). l. chlarotera nyl. 6 records; on the bark of oak, apple tree, ash, maple and poplar; 20 july 2004 (ols-l). also recorded by woźniak (1983). l. conizaeoides crombie 7 records; on the bark of linden, spruce, cherry tree, on wood and metal; 15 july 2005 (ols-l). 220 r. szymczyk and a. zalewska l. dispersa (pers.) sommerf. 5 records; on concrete; 15 july 2005 (ols-l). also recorded by woźniak (1983). l. expallens ach. 3 records; on the bark of oak, ash and poplar; 21 sept. 2005 (ols-l), (tlc 033/11). also recorded by woźniak (1983). l. hagenii (ach.) ach. 2 records; on the bark of elder and on wood; 14 may 2004 (ols-l). l. persimilis (th. fr.) nyl. 1 record; on wood; 12 aug. 2005 (ols-l 420). dd category. l. rugosella zahlbr. 1 record; on the bark of alder; 21 sept. 2005 (ols-l). l. saligna (schrad.) zahlbr. 4 records; on the bark of willow and on wood; 3 sept. 2001 (ols-l 298). also recorded by woźniak (1983). l. symmicta (ach.) ach. 5 records; on the bark of plum tree, willow, cherry tree, on wood; 20 sept. 2005 (ols-l 334). l varia (hoffm.) ach. 4 records; on the bark of oak and on wood; 20 july 2005, (ols-l 294). also recorded by woźniak (1983). lecidella elaeochroma (ach.) choisy 12 records; on the bark of elder, apple tree, horse-chestnut, maple, hazel, linden, alder, poplar, willow, on wood; 7 aug. 2001 (ols-l 349). also recorded by woźniak (1983). l. flavosorediata (vězda) hertel & leuckert 1 record; on the bark of poplar; 20 july 2004, 10 june 2006; rev. m. kukwa (tlc 033/10) (ols-l 321, 680). l. scabra (taylor) hertel & leuckert 2 records; on roof tiles; nt category; 14 sept. 2005, det. m. kukwa (tlc 2/2007/15,16) (ols-l 417). l. stigmatea (ach.) hertel & leuckert 2 records; on concrete; 29 sept. 2005 (ols-l 295). lepraria incana (l.) ach. 7 records; on the bark of oak, ash, linden and alder, on wood; 20 july 2004 (olsl 313, 314, 318). l. vouauxii (hue) r. c. harris 2 records; on the bark of ash and wood; 10 july 2004; det. m. kukwa (tlc 16/730) (ols-l 322, 323). *lichenoconium erodens m. s. christ. & d. hawksw. et d. hawksw. 2 records; on thalli of lecanora sp. growing on wood, 10 july 2004 (ols-l). lichens in the rural landscape 221 *l. xanthoriae m.s. christ. 1 record; on apothecia of xanthoria parietina growing on willow, 11 oct. 2005 (ols-l 392). *lichenodiplis lecanorae (vouaux) dyko & d. hawksw. 2 record; on apothecia of lecanora chlarotera growing on ash; 25 dec. 2004 (ols-l 342). melanelixia subargentifera (nyl.) o. blanco et al. 1 record; on ash bark; 9 may 2005 (ols-l 351). vu category, p. m. subaurifera (nyl.) o. blanco et al. 1 record; on the bark of alder; 9 may 2005 (ols-l 354). p. melanohalea exasperatula (de not.) o. blanco et al. 16 records; on the bark of oak, apple tree, ash, alder, poplar, willow, cherry tree, on wood; 20 sept. 2005 (ols-l 333). p. also recorded by woźniak (1983). micarea denigrata (fr.) hedl. 4 records; on wood; 10 june 2005 (ols-l 374). m. misella (nyl.) hedl. 1 record; on wood; 25 dec. 2005 (ols-l 287). m. prasina fr. 1 record; on wood; 15 aug. 2005 (ols-l 490), (tlc 038/1). mycoblastus fucatus (stirt.) zahlbr. 1 record; on the bark of plum tree; 10 june 2005 (ols-l), (tlc 038/2). neofuscelia loxodes (nyl.) essl. 4 records; on roof tiles; 20 aug. 2005 (ols-l 387). p. parmelia saxatilis (l.) ach. 2 records; on the bark of oak and poplar; 14 july 2005 (field record, not collected), p. p. sulcata taylor 21 records; on the bark of oak, apple tree, ash, maple, linden, alder, poplar, willow, cherry tree, on wood, 9 may 2005 (field record, not collected). also recorded by woźniak (1983). parmelina tiliacea (hoffm.) hale 15 records; on the bark of oak, apple tree, ash, linden, alder, poplar, 14 july 2005 (ols-l 353). vu category, p. also recorded by woźniak (1983). pertusaria albescens (huds.) m. choisy & werner 3 records; on the bark of ash, maple and linden; 14 july 2005 (ols-l 380). also recorded by woźniak (1983). p. amara (ach.) nyl. 7 records; on the bark of ash, oak and linden; 14 july 2005 (field record, not collected). also recorded by woźniak (1983). phaeophyscia orbicularis (neck.) moberg 13 records; on the bark of elder, apple tree, ash, poplar, willow, on concrete; 25 dec. 2005 (ols-l 397). also recorded by woźniak (1983). 222 r. szymczyk and a. zalewska phlyctis argena (ach.) flot. 10 records; on the bark of oak, ash, maple, poplar and willow; 14 july 2005 (olsl 337). also recorded by woźniak (1983). physcia adscendens (fr.) h. olivier 5 records; on the bark of oak, apple tree, ash and poplar; 14 july 2005 (ols-l 338). also recorded by woźniak (1983). ph. caesia (hoffm.) fürnr. 4 records; on boulders and concrete; 14 july 2004 (ols-l). also recorded by woźniak (1983). ph. dubia (hoffm.) lettau 4 records; on the bark of apple tree and ash, on metal; 20 sept. 2005 (ols-l 289). ph. stellaris (l.) nyl. 4 records; on the bark of elder, apple tree and plum tree; 20 sept. 2005 (ols-l 302, 357). ph. tenella (scop.) dc. 30 records; on the bark of elder, oak, apple tree, ash, horse-chestnut, maple, alder, rose, plum tree, poplar, willow, cherry tree, on wood and metal; 6 march 2005 (ols-l 331). physconia enteroxantha (nyl.) poelt 10 records; on the bark of oak, apple tree, ash, poplar, willow, on wood; 14 july 2005 (ols-l 367). also recorded by woźniak (1983). ph. grisea (lam.) poelt 5 records; on the bark of ash, poplar and willow; 14 july 2005 (ols-l). ph. perisidiosa (erichsen) moberg 3 records; on the bark of elder, ash and poplar; 14 july 2005 (ols-l 340). en category. piccolia ochrophora (nyl.) hafellner 1 record; on the bark of willow; 10 aug. 2005 (ols-l 381). vu category. placynthiella icmalea (ach.) coppins & p. james 2 records; on wood; 25 dec. 2005 (ols-l 285). pleurosticta acetabulum (neck.) elix & lumbsch 4 records; on the bark of oak and ash; 14 july (ols-l 311). en category, p. also recorded by woźniak (1983). *polysporina lapponica (schaer.) degel. 2 records; on thalli of acarospora fuscata growing on roof tiles; 12 july 2005, det. d. kubiak (ols-l 371, 372). nt category. porpidia soredizodes (lamy ex nyl.) j. r. laudon 3 records; on roof tiles; 14 sept. 2005 (ols-l 417). protoparmelia hypotremella van herk, spier & v. wirth 1 record; on wood; 9 may 2005, rev. m. kukwa (tlc 028/18) (ols-l 306). lichens in the rural landscape 223 protoparmeliopsis muralis (schreb.) m. choisy 6 records; on cement-asbestos roofing material, wood, roof tiles and concrete; 14 july 2005 (field record, not collected). pseudevernia furfuracea (l.) zopf 5 records; on the bark of oak and ash, on wood; 12 july 2005 (field record, not collected). p. also recorded by woźniak (1983). pycnora sorophora (vain.) hafellner 2 records; on wood; 20 may 2004 (ols-l 341). ramalina farinacea (l.) ach. 14 records; on the bark of oak, ash and linden; 20 aug. 2005 (ols-l 393). vu category, p. also recorded by woźniak (1983). r. fastigiata (pers.) ach. 10 records; on the bark of oak, ash and linden; 20 aug. 2005 (field record, not collected). en category, p. r. fraxinea (l.) ach. 8 records; on oak and the bark of ash; 20 aug. 2005 (ols-l 394). en category, p. also recorded by woźniak (1983). rhizocarpon distinctum th. fr. 3 records; on roof tiles; 3 july 2006 (ols-l 533). rinodina exigua (ach.) gray 1 record; on the bark of willow; 3 aug. 2005 (ols-l 299). vu category. scoliciosporum chlorococum (graewe ex stenh.) vĕzda 11 records; on the bark of elder, oak, hazel, alder, plum tree, willow, cherry tree, on wood; 20 sept. 2005 (ols-l 301). s. umbrinum (ach.) arnold 2 records; on roof tiles; 10 aug. 2005 (ols-l 513). strangospora pinicola (a. massal.) körb. 3 records; on wood; 20 aug. 2005 (ols-l). lc category. thelomma ocellatum (körb.) tibell 1 record; on wood; 6 june 2005 (ols-l 390). trapeliopsis flexuosa (fr.) coppins & p. james 1 record; on wood; 26 march 2005 (ols-l 305). t. granulosa (hoffm.) lumbsch 2 records; on wood; 20 aug. 2004 (ols-l 293). t. pseudogranulosa coppins & p. james 1 record; on wood; 10 aug. 2005 (ols-l). verrucaria muralis ach. 1 record; on concrete; 10 aug. 2005 (ols-l 384). v. nigrescens pers. 2 records; on concrete; 10 aug. 2005 (ols-l 383). *vouauxiella lichenicola (lindsay) petr. & syd. 1 record; on apothecia of lecanora sp. [l. subfusca-group] growing on oak; 12 july 2005 (ols-l). 224 r. szymczyk and a. zalewska xanthoria candelaria (l.) th. fr. 9 records; on the bark of oak, ash, horse-chestnut, maple, alder, willow, on metal; 20 sept. 2005 (ols-l 329). also recorded by woźniak (1983). x. elegans (link.) th. fr. 2 records; on concrete; 14 july 2005 (field record, not collected). x. parietina (l.) th. fr. 12 records; on the bark of elder, apple tree, ash, maple, poplar, willow, on concrete and cement-asbestos roofing material; 25 oct. 2005 (ols-l 392). also recorded by woźniak (1983). x. polycarpa (hoffm.) rieber 14 records; on the bark of elder, oak, apple tree, ash, rose, plum tree, poplar, cherry tree, willow, on metal; 20 sept. 2005 (ols-l 370). also recorded by woźniak (1983). *xanthoriicola physciae (kalchbr.) d. hawksw. 1 record; on apothecia of xanthoria polycarpa growing on metal; 20 sept. 2005 (ols-l 370). ecological groups it can be assumed that the analysed biota was made up in its entirety of synanthropic lichens (fałtynowicz 1994; olech 1998) occurring in anthropogenic habitats and on man-made substrates. according to olech (1998), those lichens can be divided into three groups. the first group comprises macroautoapophytes (euapophytes) which are more likely to inhabit anthropogenic habitats than natural ecosystems. this group was represented, among others, by the following taxa: amandinea punctata, anaptychia ciliaris, melanohalea exasperatula, lecanora varia, parmelina tiliacea, phaeophyscia orbicularis, physcia dubia, ramalina fastigiata, r. fraxinea, xanthoria parietina and x. polycarpa. in the investigated area, they were noted mostly on the bark of roadside trees or fruit trees. most of them are coniophilous and nitrophilous lichens. they show a preference for habitats with ample sunlight and are resistant to draughts (barkman 1958, wirth 1995). the observed specimens had well-developed thalli and often produced numerous ascomata (e.g. anaptychia ciliaris and the listed species of the genus ramalina). the second group of synanthropic lichens consists of mesoautoapophytes (hemiapophytes) which are equally often found in anthropogenic habitats and in natural ecosystems, among them acarospora fuscata, buellia griseovirens, cladonia coniocraea, evernia prunastri, lecidella elaeochroma, parmelia saxatilis, pertusaria amara and rhizocarpon distinctum. the least populous group were the microautoapophytes which inhabit anthropogenic habitats only sporadically (olech 1998). in the studied area, they were: bacidina phacodes, chaenotheca trichialis, coenogonium pineti, micarea misella and m. prasina. the first taxon grows on a varied range of substrates, but it is rarely noted outside of wooded areas (cieśliński 2003, fałtynowicz 2003). the remaining species are believed to be typical of forest habitats, and they are most often found in damp and shaded microniches (cieśliński 2003, czarnota 2007). in the investigated area, chaenotheca trichialis, micarea misella and m. prasina were noted on dead wood at lichens in the rural landscape 225 the base of old fences or tree stumps surrounded by dense herbaceous vegetation. coenogonium pineti was observed on tree bark, on lower parts of the trunk of alnus glutinosa by the side of the river. this group of lichens is also inclusive of anisomeridium polypori which was also found on the bark of ash, alder and willow trees. until recently, this species was reported almost exclusively from forest communities in poland (fałtynowicz 2003), whereas other european sources (e.g. earland-benett 1994; wirth 1997) claim that this species is able to colonize shaded microhabitats near human settlements. protected, threatened, rare and otherwise interesting species the area of the nowica village is a habitat of many protected and threatened lichens. from among the 96 noted species, 14 are legally protected and 18 are threatened taxa in poland (cieśliński et al. 2006). the highest local frequency was observed in respect of: parmelina tiliacea (vu, p), ramalina farinacea (vu, p), r. fastigiata (en, p) and r. fraxinea (en, p). the lichen biota of the nowica region also features interesting species considered rare both in poland and in gdańsk pomerania. the group of taxa with a few localities is inclusive of bacidia adastra, caloplaca obscurella, chaenothecopsis savonica and lecanora persimilis. the first one has been recently described (sparrius and aptroot 2003) , and to date its only recorded localities in poland are the city of olsztyn (kubiak and sparrius 2004; kubiak 2005), the arboretum in rogów (kubiak and szczepkowski 2006) and three sites in the region of gdańsk pomerania (kukwa 2006). in the area of the nowica village, bacidia adastra was found on the bark of an old apple tree. the remaining species have very fine thalli that are difficult to spot during field observations which could explain the small number of localities recorded in poland. until recently, the most frequently observed species was caloplaca obscurella (see fałtynowicz 2003), found on the bark of a roadside willow. chaenothecopsis savonica was noted only in the puszcza białowieska forest (sparrius 2003), in the elbląg high plain adjacent to the warmia plain (szymczyk 2007), as well as in the puszcza borecka forest and in the iława lakeland (zalewska – unpubl. materials). in the investigated area, the species was found on an old wooden fence. lecanora persimilis has been recorded in poland at three older localities (fałtynowicz 2003) and at several subsequent localities (jando and kukwa 2002; czyżewska 2003a; łubek and cieśliński 2004; kubiak 2005; kukwa 2006). in the studied area, it was observed on the exposed wood of an old roadside ash tree. species which are rarely found on the regional scale are: anaptychia ciliaris, bacidina phacodes, parmelina tiliacea, piccolia ochrophora, polysporina lapponica, protoparmelia hypotremella, pycnora sorophora, thelomma ocellatum and xanthoria elegans (fałtynowicz and kukwa 2003, 2006). the most intriguing discovery was that of lecidella scabra, a montane epilithic lichen found on old roof tiles in nowica. this lichen was believed to be extinct in gdańsk pomerania (fałtynowicz and kukwa 2003), but it was recently discovered at several localities on the stone walls of historical buildings (fałtynowicz and kukwa 2006; m. kukwa, pers. comm.). in europe, this species was also noted on similar, anthropogenic substrates (brightman and seaward 1977; gilbert 1990). 226 r. szymczyk and a. zalewska lichens growing on corroded metal were another interesting group of species. the study revealed 7 taxa that sometimes occur on this type of substrate (brightman and seaward 1977; gilbert 1990): caloplaca holocarpa, candelariella aurella, c. vitellina, lecanora conizaeoides, physcia dubia, ph. tenella, xanthoria candelaria and x. polycarpa. those lichens were observed on the metal parts of old farm machines, such as the plough and the trailer. discussion although the village of nowica covers a small area, the presented lichen species could be a model example of lichen biota in rural areas with traditional, low-intensity farming systems. there are no large animal farms in the village, its immediate vicinity and surroundings, which could drastically raise the local levels of ammonia and other nitrogen compounds that contribute to the expansion of nitrophilous lichens and to the massive elimination of acidophilic species (see benfield 1994; van herk 1999; 2002; wolseley et al. 2006a). in the majority of farms the stocking density is low or moderate (up to 20 animals), grasslands occupy large areas and arable land is not sprayed with slurry. similarly to other areas of this type, the eutrophication of tree bark, wooden fences and other substrates leads to the emission of ammonia from decomposing manure stored by farm buildings or used as fertilizer. outside the reach of ammonia gas (up to 300 m from the source of emission, see the works of m. a. sutton et al., as cited by wolseley et al. 2006a), chemical changes in the surface of the substrate are caused by the sedimentation of dust containing ammonia derivatives (e.g. ammonium sulfate or nitrate) precipitated in the form of dry or wet deposition as well as chemical fertilizers as nitrates and phosphates. it should also be noted that industrial, municipal and traffic-related air pollution, which has an opposing effect on the substrate and directly eliminates sensitive species of fruticose and foliose lichens (hawksworth and rose 1970; gries 1996), does not have a significant effect on the investigated lichen biota. vehicle traffic on roads intersecting nowica is not very intense, and the main sources of sulfur dioxide and nitrogen oxide emissions from the combustion process could be the elbląg combined heat and power plant and the city of elbląg situated 25 km to the west. yet owing to the distance and the low level of pollution (advanced filtering system, biogas combustion), the plant and the city of elbląg have a small environmental impact. direct measurements have not been performed in nowica, but in view of the available data, the mean annual concentration of so2 and no2 in ambient air in the county of braniewo is among the lowest in the province of warmia and mazury (mean annual maxima: so2 – 6.7 μg/m 3, no2 – 5.1μg/m 3, acc. to jamiołkowski 2002). in 2003-2005, mean annual so2 concentrations in elbląg reached 2 μg/m 3 (the allowable level in 2005 was 20 μg/m3), while mean annual no2 concentrations were in the range of 20-26 μg/m3 (the allowable level in 2005 was 50 μg/m3) (kacprzykchynczewska 2006). available data suggest that the investigated area is characterized by an average level of substratum eutrophication and a low level of gaseous pollution. under such conditions, repeatable distribution patterns of epiphytic species were observed on free-standing and roadside trees. the predominant species occurring on the bark of deciduous trees were fruticose lichens, mainly evernia prunastri and ramalina lichens in the rural landscape 227 farinacea. the following species were less abundant: ramalina fraxinea, r. fastigiata, the foliose physconia enteroxantha, physcia tenella, phaeophyscia orbicularis, parmelia sulcata, melanohalea exasperatula, as well as species with crustose thalli, mostly pertusaria albescens, p. amara, phlyctis argena and small specimens of lecanora carpinea, l. chlarotera and l. argentata. in addition to ramalina spp. and physcia s.l., the group of nitrophilous lichens was also inclusive of xanthoria parietuna, x. polycarpa, amandinea punctata, candelariella reflexa and c. xanthostigma. other species, e.g. anaptychia ciliaris, pleurosticta acetabulum and smaller specimens of melanelixia subargentifera, physcia stellaris and physconia perisidiosa occurred less frequently, but in the form of large thalli. interesting, relatively unexplored members of those lichen communities were caloplaca obscurella, l. flavosorediata, piccolia ochrophora and rinodina exigua which are found in other areas of northern poland. in future studies, attention should be also paid to protoparmelia hypotremella and strangospora pinicola, often found on roadside trees (zalewska – unpublished materials), which were observed on wood in the investigated area. the study area is distinguished by relatively frequent and abundant populations of parmelina tiliacea. according to cieśliński (2003), this epiphyte is still widespread in the old prussian lowland adjacent to the warmia plain, but it has a decreasing share of the mesoregions situated further to the east. the high frequency of candelariella reflexa is another specific feature of the investigated lichen biota. this taxon is less commonly identified and generally believed to be less widespread than the similar c. xanthostigma (fałtynowicz 1992; cieśliński 2003). epiphytes growing on roadside and free-standing trees are one of the most endangered ecological lichen groups in poland (czyżewska 2003b). their populations have been most drastically reduced in industrialized areas with the greatest air pollution, in particular in the lowland areas of upper silesia and opole silesia. communities which are most typically developed and least threatened by toxic compounds are still found in north-eastern poland (czyżewska 2003b; cieśliński 2003). the felling of roadside trees during road construction poses one of the fastest growing threats for this ecological group, even in the above area. such projects are initiated “to improve the safety” of roads which are not regularly upgraded. despite frequent protests of ecologists, this argument is frequently abused by the authorities. final remarks in view of growing pressure, research studies are needed to investigate the condition of and changes in epiphytic communities in rural areas. observations of lichens growing on other anthropogenic substrates will provide complementary information on the hemerophily of many species and will validate data relating to lichens that, until now, have been associated primarily with forest communities, as discussed in this study. such studies should cover the broadest possible spectrum of substrates and rarely investigated wet and shaded microhabitats. the conducted study has also shown that open areas can be a habitat of rare species whose distribution in poland remains relatively unknown. to conclude, rural areas which are characterized by a low level of substrate eutrophication and which are not affected by air pollution provide habitat for potentially valuable and interesting lichen biota. the structure of the lichen biota of the 228 r. szymczyk and a. zalewska village of nowica can be used as a contemporary model and a reference point for other studies of this type. in the future, the collection of larger quantities of data will support more advanced analyses similar to the research of ruderal vascular flora (see wołkowycki 2000). acknowledgements. the authors would like to thank martin kukwa, ph.d., for verifying the determinations and for his critical evaluation of this paper. we would also like to thank two anonymous reviewers for their comments and improvements to the draft manuscript. in the course of this study, the first author received a grant from the european social fund and the central budget as part of the integrated regional development operational program 2006-2007. references barkman j. j. 1958. phytosociology and ecology of cryptogamic epiphytes. van gorcum & comp., assen. 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[wolseley et al. 2006b]. variation of lichen communities with landuse in aberdeenshire, uk. li-lichenologist 38(4): 307-322. wołkowycki d. 2000. różnicowanie i ujednolicanie się flor ruderalnych w warunkach izolacji środowiskowej. monogr. bot. 87: 1-163. woźniak i. 1983. porosty niziny warmińskiej. master thesis. zakład taksonomii roślin uniwersytetu im. adama mickiewicza w poznaniu (department of plant taxonomy, adam mickiewicz university in poznań), poznań, 83 pp. porosty w krajobrazie rolniczym na równinie warmińskiej s t r e s z c z e n i e praca prezentuje wyniki badań lichenologicznych, których celem było zarejestrowanie gatunków porostów w krajobrazie rolniczym wsi nowica na równinie warmińskiej (fig. 1). plechy porostów zbierano ze wszystkich substratów antropogenicznych. przedstawiono 104 taksony, w tym 96 gatunków porostów i 8 gatunków grzybów niezlichenizowanych. spośród odnotowanych grzybów zlichenizowanych – 14 to gatunki prawnie chronione, a 18 znajduje się na czerwonej liście porostów zagrożonych. biota analizowanego terenu zawiera gatunki rzadkie w polsce, notowane na pojedynczych lub nielicznych stanowiskach, np. bacidia adastra, caloplaca obscurella, chaenothecopsis savonica i lecanora persimilis. zarejestrowano także stanowisko lecidella scabra, do niedawna uważanego za wymarły na pomorzu gdańskim. listę gatunków w całości stanowią porosty hemerofilne, zasiedlające siedliska i substraty wytworzone przez człowieka. analizowana biota może stanowić model składu gatunkowego porostów na terenach rolniczych o niewielkim poziomie eutrofizacji podłoży, położonego poza zasięgiem oddziaływania zanieczyszczeń powietrza. 2014-01-01t11:48:20+0100 polish botanical society 2014-08-20t21:45:07+0200 polish botanical society 2014-08-20t22:53:57+0200 polish botanical society 2014-08-20t22:13:03+0200 polish botanical society cephalotrichum stemonitis as a biofilm inhabitant in the gold mine in poland andrzej chlebicki department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, ibchlebick@ib-krakow.pl chlebicki a.: cephalotrichum stemonitis as a biofilm inhabitant in gold mine in poland. acta mycol. 43 (1): 67–70, 2008. cephalotrichum stemonitis and its synanamorph echinobotryum atrum isolated from bacterial biofilm is presented. key words: mine fungi, cephalotrichum, biofilm, distribution introduction dry-spored synnematous anamorphs from the form-genus cephalotrichum link are the asexual states of microascaceae. however so far a teleomorph state is known for no species of this genus (abbott 2000). microascaceae include five genera: microascus, kernia, petriella, pseudoalescheria and lophotrichus (abbott et al. 2002) as well as eight anamorphic genera containing about 50 species (abbott 2000). abbott (2000) analyzed the subunit 18s rdna for 34 taxa of microascaceae. he obtained microascus clade consisting of microascus longirostris, m. nidicola, m. cirrhosus, m. trigonosporus, cephalotrichum stemonitis, wardomyces anomalus, cephalotrichum cylindricum and microascus brevicaulis. it indicate that the genus microascus can be recognized as hypothetical teleomorph state for cephalotrichum stemonitis. in genbank there are available 3 sequences of ribosomal rna genes (5.8s rrna, 28s rrna and large subunit rrna) of doratomyces stemonitis (in fact cephalotrichum stemonitis) with access numbers: af400852, ef029213 and aj608983. synnemata of cephalotrichum producing chains of powdery conidia are relatively large with ‘bootle brush’ or “feather” appearance. ovoid conidia are produced from annelidic conidiogenous cells covering sporogonous area. cephalotrichum synnemata are adapted for dispersion not only by air currents but also by insects (abbott 2000, 2002). habitat of cephalotrichum fungi includes soil, compost, wood, wood treated with fungicide, herbaceous stems, oat seeds, decaying plant material and dung, finger nail, sawdust and straw used for growing shiitake, airborne contaminant of wheat-straw agar plate, egg of gypsy moth (lymantaria dispar), roots of potato, acta mycologica vol. 43 (1): 67–70 2008 68 a. chlebicki basidiomycete detritus, manore pile, bronchial washing, right feel, leaves of needleleafed tree (abbott 2000). c. stemonitis was noted in coyote and rat dung, indoor air of honeybee (apis mellifera) overwintering facility, cone of white spruce, sandy soil, decayed wood of white spruce, soil of elm woods, agricultural soil. it is also important colonizer of ppvc buried in soil (sabev et al. 2006). these authors mentioned it as doratomyces spp. in the article, but in genbank it is more precisely determined as d. stemonitis. malloch and hubart (1987) described unnamed species of microascus from ramoil cave. it means that fungi from this group can inhabit also underground environments. fassatiova (1970) noted c. stemonitis on wood in uranium mine in czech republic. study area and methods gray biofilms of rock-inhabiting bacteria in gertruda adit in a closed gold mine located in złoty stok in lower silesia were chosen for microbial analysis. this mine posseses constant conditions such as low temperature ca 10º c, darkness, high humidity, high as concentration and other toxic substances (chlebicki et al. 2005). so far 12 species of fungi were noted in this mine (chlebicki et al. 2005; chlebicki, lorenc 2006). bacteria with fungi were collected in sterile plastic tubes and refrigeratorated at 10º c. fungal growth was performed on drbc, rbc, yma and pda media. inoculated media on petri dishes were putted in incubator at 10º c. the morphological characters of the living fungi were examined in water and cotton blue in lactophenol using light microscopy (nikon smz 1500, nikon labophot 2 and nikon eclipse 800). microphotographs were taken with these microscopes equipped with a digital camera. for scanning electron microscope (sem) studies mycelium was coated with gold, and photographed using a leo 1430 vp zeiss microscope with a working distance of ca 10 mm. fungus-species nomenclature follows abbott (2000). results cephalotrichum stemonitis (pers.) nees magazin ges. naturf. freunde, berlin 3: 20 (1809) synonymy: doratomyces stemonitis (pers.) f.j. morton & g. sm., mycol. pap. 86: 70, 1963 doratomyces stemonitis var. keratinolyticus (dominik & majchr.) dominik & majchr. ekol. pol., ser. a 18: 603, 1970 echinobotryum atrum corda, sturm’s deutschl. flora, iii (pilze) 3(12): 51 (1829) periconia stemonitis pers., syn. meth. fung. (göttingen): 687 (1801) stysanus stemonitis (pers.) corda, icon. fung. (prague) 1: 22 (1837) description: mycelium creamy-white to dark brown, after 7 days on pda 17,5-21 mm diam., on rbc 17,0 mm diam., and on drbc 13-17 mm, all in room temperature (fig. 1a, b, d). first conidiophores of synanamorph – echinonobotryum atrum – appeared throughout the mycelium after 12 days on pda. mycelium growing on rbc after two weeks produced rings of densely distributed synnemata of c. stemonitis in central part of the mycelium. mycelium on pda formed at the beginning e. cephalotrichum stemonitis 69 atrum throughout the surface of dark brown colony and later numerous synnemata of c. stemonitis. mycelium on yma form brown and irregular colony patches (fig. 1c). synnemata 190-300 μm long, sterile part 130-210 μm, fertile part 60-120 μm long, (irregular heads) similar to feather (fig. 2b, c) associated with synanamorph echinobotryum atrum (fig. 2a), conidiophores synnematous, brown, conidiogenous cells ampulliform, percurrent, conidia smooth (fig. 2d), ovoid with pointed apex and truncate base 7-9,1 x 4,2-6,1 μm, pale brown. material examined: on rock bacterial biofilm in gertruda adit in gold mine in złoty stok, lower silesia, poland, 16 october 2006, coll. a. chlebicki. first culture obtained on drbc medium, then transferred to other media. discussion so far two species of the genus cephalotrichum link were noted in poland. dominik and majchrowicz (1965) and dominik (1970) described a new variety doratomyces stemonitis var. keratinolyticus (dominik & majchr.) dominik & majchr. on the basis of specimen isolated from the soil. conidia of this specimen are something smaller than typical variety, 4-6 x 2.5-4 μm and ‘very slightly rough’ (dominik 1970). it resembles cephalotrichum microsporum (sacc.) p. m. kirk. moreover these authors did not mention the presence of echinobotryum synanamorph which is diagnostic character of cephalotrichum stemonitis. the next species cephalotrichum putredinis (corda) s. p. abbott was reported by dominik (1970) as doratomyces albus (szilvinyi) dominik. unfortunately, these collections are not available for investigation. presence of cephalotrichum stemonitis in bacterial biofilm is accidental. however, as indicate informations of fassatiova (1970) and malloch and hubart (1987), such fungi were noted in subterranean environments. mille-lindblom (2005), hogan and kolter (2002) and kirkwood (2002) noted mostly antagonistic relation between fungi and bacteria. fungi were always negatively affected by presence of bacteria. penetration of fungal hyphae of fusarium oxysporum was not observed where microcolonies of pseudomonas were present, moreover pseudomonas bacteria attached and colonized fungus hyphae (bolwerk et al. 2003). also growth of cephalotrichum stemonitis was suppressed by bacteria from the genus pseudomonas isolated from biofilm. references abbott s. p. 2000. holomorph studies of the microascaceae (phd dissertation). edmonton, alberta: univ. alberta. 196 pp. abbott s. p. 2002. insects and other arthropods as agents of vector-dispersal in fungi. www.precisionenv. com./pdfs/abbottinsectdispersal.pdf abbott s. p., lumley t. c., sigler l. 2002. use of holomorph characters to delimit microascus nidicola and m. soppii sp. nov., with notes on the genus pithoascus. mycologia 94 (2): 362–369. bolwerk a. lagopodi a. l., wijfjes a. h. m., lamers g. e. m., chin-a-woeng t. f. c., lugtenberg b. j. j., blomberg g. v. 2003. interaction in the tomato rhizosphere of two pseudomonas biocontrol strains with the phytopathogenic fungus fusarium oxysporum f. sp. radicis-lycopersici. mol. plantmicrobe interact. 16: 983–993. chlebicki a., godzik b., lorenc m. w., skłodowska a. 2005. fungi and arsen-tolerant bacteria in the hypogean environment of an ancient gold mine in lower silesia sw poland. polish bot. stud. 19: 81–95. 70 a. chlebicki chlebicki a., lorenc m. w. 2006. the troglophile fungus, physisporinus vitreus on a mine wall at złoty stok, poland. polish. bot. j. 22: 149–154. dominik t. 1970. observations of new or noteworthy fungi from region of szczecin. zeszyty naukowe wyższej szkoły rolniczej w szczecinie 32: 71–108. dominik t., majchrowicz i. 1965. second contribution to the knowledge of keratynolytic and keratyno-second contribution to the knowledge of keratynolytic and keratynophilic soil fungi in the region of szczecin. ekologia polska 13: 415–447. fassatiova o. 1970. micromycetes inhabiting the mines of přibram (czechoslovakia). česka mykol. 24: 162–165. hogan d. a., kolter r. 2002. pseudomonas-candida interactions: and ecological role of virulence factors. science 296: 2229–2232. kirkwood m. l. 2002. bacteria-fungi interactions: pathogenesis meets ecology. trends in microbiology 10: 397–398. malloch d., hubart j. m. 1987. an undescribed species of microascus from the cave of ramoiul. canad. j. bot. 65: 1281–1283. mille-lindblom c. 2005. interaction between bacteria and fungi on aquatic detritus-causes and conse-interaction between bacteria and fungi on aquatic detritus-causes and consequences. acta universitatis upsaliensis. digital comprehensive summaries of uppsala dissertations from the faculty of science and technology 46. 42 pp., uppsala. sabev h. a, handley p. s., robson g. d. 2006. fungal colonization of soil-buried plasticized polyvinyl chloride (ppvc) and the impact of incorporated biocides. microbiology 152: 1731–1739. cephalotrichum stemonitis zasiedlający bakteryjny biofilm w kopalni złota w polsce s t r e s z c z e n i e praca zawiera opis grzyba cephalotrichum stemonitis i jego synanamorfy echinobotryum atrum wyizolowanych z bakteryjnego biofilmu z sztolni gertrudy w kopalni złota w złotym stoku. pierwsza informacja o tym gatunku podana z polski przez t. dominika i i. majchrowicz jest niezbyt dokładna. diagnostyczną cechą gatunku c. stemionitis jest obecność synanamorfy e. atrum o czym wymienieni autorzy nie wspominają. podano również podstawowe dane o siedliskach i ekologii tego gatunku. izolacja tego rzadkiego gatunku grzyba z bakteryjnego biofilmu nie była dotychczas notowana. fig. 1. colonies of cephalotrichum stemonitis on different media: a – on rbc; b – on drbc; c – on yma; d – on pda. fig. 2. morphology of cephalotrichum stemonitis and its synanamorph – echinobotryum atrum: a – conidia of e. atrum; scale bar = 10 µm; b – synnemata of c. stemonitis on drbc medium; c – synnemata of c. stemonitis similar to feather; d – conidies of c. stemonitis. 2014-01-01t11:47:18+0100 polish botanical society some interesting species of the genus ascochyta ewa połeć and małgorzata ruszkiewicz-michalska department of mycology, university of łódź, banacha 12/16 pl-90-237 łódź, ewa_polec@op.pl połeć e., ruszkiewicz-michalska m.: some interesting species of the genus ascochyta. acta mycol. 46 (2): 187–200, 2011. the paper presents eleven species of ascochyta recently collected in central and southern part of poland. two of them, ascochyta bondarceviana melnik and ascochyta equiseti (desm.) grove noted in poland for the first time, are illustrated with microphotographs. in addition, nine other species are newly reported on their host plants species in the country. short characteristics of the fungi species based on the collected specimens and the distribution maps of all fungi taxa are presented. key words: micromycetes, anamorphic fungi, phoma, coelomycetes, plant parasites, poland introduction the genus ascochyta, one of the largest (over 1 400 names) and the most important genera of anamorphic fungi, was often examined since its description by libert in 1830. as a result, many species were reallocated to other genera at the beginning of 20th century, and some of them (e.g., ascochytella tassi and ascochytula died.) were later re-synonymized with ascochyta (buchanan 1987). the great interest in this genus stems from the fact that its members are the parasites of numerous cultivated and wild plants, causing diseases of economically important taxa (melnik 2000). ascochyta anamorphs are characterized by unilocular, glabrous, ostiolate, pycnidial conidiomata and usually uniseptate, hyaline to pale-coloured conidia. the known teleomorphs are placed mainly in didymella sacc. (pleosporales) (kirk et al. 2008), and according to melnik (2000) and index fungorum (www.indexfungorum.org) some ascochyta species are also linked to two other ascomycetous genera, namely lepto­ sphaeria ces. & de not. (pleosporales) and mycosphaerella johanson (capnodiales). the genus ascochyta was the subject of the monographic studies worldwide (punithalingam 1979, 1988; melnik 2000). the 72 species recorded in poland were examined by sałata (2002) and 60 other species included in his monograph were acta mycologica vol. 46 (2): 187–200 2011 188 e. połeć and m. ruszkiewicz-michalska known from the neighbouring countries, and were thus expected to occur also in poland. due to new findings (chlebicki 2002; mułenko, wojdyło 2002; kowalski 2004; piątek, wołczańska 2004; kozłowska, mułenko 2005; ruszkiewicz-michalska 2006; wołczańska 2010) and the comprehensive literature survey (mułenko, kozłowska 2008), 60 other species have been reported from our country during the next 10 years. in total, 132 species of this genus are currently known to occur in poland, some of which were predicted by sałata (2002), e.g., a. aristolochiae sacc., a. trans­ lucens kabát & bubák and a. urticae a.l. sm. & ramsb. material and methods the analysed material has been collected mostly as a result of the systematic studies of plant parasitic micromycetes carried out in urbicoenoses of the łódź city (połeć 2010). the fresh specimens mounted in lactophenol picric acid solution (fluka) were examined using the microscopes nikon eclipse e200 and nikon eclipse 50i; measurements of the morphological structures were made also in tap water. microphotographs of morphological structures of the species new for polish mycobiota were taken with a nikon ds-f1 digital camera. the base for identification of host plants and fungi were the keys by szafer, kulczyński and pawłowski (1986) and rutkowski (2004), and taxonomic monographs by melnik (2000) and sałata (2002), respectively. the nomenclature of fungi taxa is given after mułenko and kozłowska (2008) while the nomenclature of hosts follows checklist by mirek et al. (2002). available literature data were analysed and the distribution maps of all the species listed were prepared. the vouchers representing current collections are deposited in the herbarium universitatis lodziensis (lod) in the series of parasitic fungi labeled as pf. results as a result of the studies, eleven interesting species of the genus ascochyta were identified, including two taxa new for polish mycobiota, namely ascochyta bon darceviana melnik (on ribes sp.) and a. equiseti (desm.) grove (on equisetum arvense l.), both belonging to the subgenus ascochyta. nine other species are noted for the first time on host plant in poland: ascochyta daturae sacc., a. doronici allesch., a. euphrasiae oudem., a. infuscans ellis & everh., a. lamiorum sacc., a. leonuri ellis & dearn., a. sodalis naumov, a. sonchi (sacc.) grove and a. verbascina thüm. these are classified by melnik (2000) in the ascochyta subgenus libertia melnik, characterized by consistently single, central or sometimes displaced septum. the majority of the hosts are wild plants, three species are cultivated and one, galinsoga parviflora cav., is an alien and expansive element in european flora (daisie). interesting ascochyta species 191 ascochyta doronici allesch. leaf spots circular, oval or irregular, ochraceous, brown or grey-brown. pycnidia scattered, sometimes aggregated, often confluent, immersed or erumpent, pale to dark brown, sometimes almost black, globose-depressed or lentiform, up to 125.0 μm in diam., with a circular pore, 15.0 μm in diam. conidia cylindrical or oblongellipsoidal, both ends rounded, straight or slightly bent, 1-septate, not or somewhat constricted, 10.0-12.5 x 2.5-3.75 μm. morphological features of the specimen generally correspond to those mentioned by melnik (2000). material examined. on achillea millefolium l. s. str., central poland, łódź, sielanka park, 15 july 2005, leg. m. jakiel, lod pf 2152. notes. worldwide, ascochyta doronici has been observed on leaves, stems, fruits and seeds of members of 39 genera of the asteraceae family. the distribution of the species is circumglobal (melnik 2000; farr et al. 2011). in poland (fig. 2) it is so far known on artemisia vulgaris l. from the parkowe nature reserve near złoty potok on the wyżyna częstochowska upland (ruszkiewicz-michalska 2006), on hie­ racium polonicum błocki from botanical garden in kraków (piątek, wołczańska 2004), on homogyne alpina (l.) cass. from the tatra mts (mułenko, kozłowska and sałata 2004). from more then one locality the species is known only on taraxacum officinale f. h. wigg.: from radom, słowiński national park and złoty potok on the wyżyna częstochowska upland (adamska 2001; ruszkiewicz-michalska 2006; sałata 2002; sałata, mułenko and wołczańska 1994). it has been recorded on achil­ lea millefolium l. for the first time in poland. ascochyta equiseti (desm.) grove leaf spots small, greyish white. pycnidia on stems, scattered or arranged, immersed, dark brown or black, oval or almost globose, 115.0-130.0 x 120.0-145.0 μm, with a circular pore 20.0-25.0 μm in diam. conidia oblong-ellipsoidal, oval, both ends rounded, straight, 1-septate, not or slightly constricted, 10.0-12.5 x 2.5-3.75 μm (fig. 1 c, d). according to the data of melnik (2000) pycnidia are 200.0-800.0 μm in diam. and conidia are (8.0-)10.0-16.0 x (2.5-)3.0-4.0(-4.5), while according to the characteristics of polish specimens given by sałata (2002) pycnidia are smaller, reaching 160.0-220.0(-330.0) μm in diam. and conidia measure mainly (7.0-)10.0-15.0(-18.0) x (2.5-)3.0-3.5(-4.0). material examined. on equisetum arvense l., central poland, łódź, bolesława chrobrego residential area, park, 01 oct. 2006, leg. d. papierz, lod pf 3025 (fig. 3). notes. the species has been recorded on dead and dying leaves and stems of equisetum spp. from north america (usa) and many european countries (melnik 2000; farr et al. 2011). although the species was expected by sałata (2002), it has not been observed in poland yet. ascochyta euphrasiae oudem. leaf spots circular or irregular, grey-brown or grey. pycnidia scattered, immersed, from yellowish to dark brown, globose-depressed and lentiform, 92.5-120.0 x 105.0-135.5 μm, with a circular pore, up to 20.0 μm in diam. conidia cylindrical, both ends rounded, straight, sometimes slightly bent, 1-2-septate, not or slightly constricted, 8.75-12.5 x 2.5-3.75 μm. according to the data of melnik (2000) pycnidia are up to 200.0 μm in diam. and conidia measure 7.0-12.0(-13.5) x 3.0-4.0, while according to the descriptions given interesting ascochyta species 193 ranunculoides l. in bohukały near bug river valley (danilkiewicz 1990; sałata 2002) (fig. 3). anemone sylvestris l. is a new host species for the fungus in poland. in addition, ascochytella vitalbae (briard & hariot) died. on the members of ranunculaceae (clematis recta l. and c. vitalba l.) was reported from poland. that record was classified as ascochyta indusiata bres. (mułenko, kozłowska 2008); the name is however considered to be a synonym of phoma clematidina (thüm.) boerema (boerema et al. 2004). ascochyta lamiorum sacc. leaf spots circular, oblong or irregular, yellow-brown, dark brown or grey-brown. pycnidia scattered or sometimes aggregated, immersed, light to dark brown, globose or globose-depressed, sometimes lentiform 107.8-150.0 x 110.0-172.5 μm, with a circular pore, 15.0-17.5 μm in diam. conidia predominantly cylindrical, sometimes table 1 ascochyta species associated with members of ranunculaceae (compilation based on the data given by melnik 2000 and farr et al. 2011) fungi species host species conidia (μm) distribution a sc oc hy ta s ub ge nu s a sc oh yt a a. actaeae (bres.) davis actaea alba (l.) mill., a. rubra (aiton) willd., a. spicata l., cimicifuga racemosa (l.) nutt., delphinium elatum l., hydrastis sp., thalictrum flavum l., t. minus l. 12.0-28.0 x 5.0-7.0 europe (estonia, germany, latvia, uk, poland, russia), north america (usa) a. aquilegiae (rabenh.) höhn. aquilegia spp., delphinium spp. 10.0-20.0 x (3.0-)4.0-6.0 europe (austria, belarus, bulgaria, former czechoslovakia, france, germany, hungary, italy, poland, russia), asia (armenia), north america (usa), south africa (zimbabwe) a sc oc hy ta s ub ge nu s l ib er tia a. aconitana melnik aconitum moldavicum hacq. 10.0-15.0(-18.0) x 4.5-6.3 europe (romania) a. dolomitica kabát & bubák atragene sibirica l., clematis alpina (l.) mill., clematis spp., hepatica nobilis schreb., ranunculus thora l. (10.0-)13.0-20.0 (-22.0) x 3.0-5.0 europe (austria, former czechoslovakia, germany, latvia, poland, romania), asia (russia, kazakhstan) a. infuscans ellis & everh. anemone nemorosa l., a. ranunculoides l., clematis vitalba l., helleborus odorus waldst. & kit, pulsatilla vulgaris mill., ranunculus abortivus l., ranunculus sp. 8.0-15.0(-16.0) x 3.0-4.5 europe (bulgaria, former czechoslovakia, poland, russia), north america (canada, usa) a. patagonica speg. aconitum septentrionale koelle, a. lycoctonum l. emend. koelle subsp. lycoctonum, aconitum sp., anemone riparia fernald, anemone sphenophylla poepp., aquilegia sp. (6.5-)8.0-10.0 x 2.5-4.0 europe (bulgaria, hungary, russia), north america (canada), south america (argentina) a. savulescui rădul. & negru thalictrum minus l. 18.0-26.0 x 7.0-10.0 europe (romania) a. vitalbicola maire clematis vitalba l. 15.0-18.0 x 5.0-6.0 europe (bulgaria, spain) interesting ascochyta species 195 diam., surrounded by small dark cells. conidia cylindrical, both ends rounded, straight or slightly flexuous, 0-1-septate, slightly constricted, 10.0-16.25 x 2.5-3.75 μm. morphological features of the specimen generally correspond to those given by melnik (2000). material examined. on mentha x citrata ehrh. subsp. citrata, central poland, łódź, botanical garden, sect. of medicinally and industrially important plants, cultivated, 28 aug. 2004, leg. e. połeć, lod pf 3134. notes. the species has been reported on lamiaceae members: lamium macula­ tum l., leonurus cardiaca l., mentha arvensis l., m. longifolia (l.) l., nepeta cataria l., n. mussinii spreng. ex henckel and n. pannonica l. from asia, north america and many european countries (melnik 2000; farr et al. 2011). in poland (fig. 4) it is known to occur on mentha arvensis l. in firlej near radom (sałata et al. 1994; sałata 2002), mentha x verticillata l. in białowieża national park (mułenko 1996; faliński, mułenko 1997), nepeta cataria l. var. citriodora and nepeta sp. in zakrzów near koźle (miczyńska 1966; sałata 2002). as mentha x citrata ehrh. subsp. citrata has not been listed among the species infected with ascochyta leonuri (melnik 2000; farr et al. 2011) it is presumably a new host of the fungus. according to boerema et al. (2004) ascochyta nepetae is a synonym of phoma nepeticola (melnik) dorenb. & gruyter. ascochyta sodalis naumov = ascochyta plantaginis sacc. & speg., a. plantaginicola melnik leaf spots oval or circular, sometimes irregular, yellowish or brown. pycnidia immersed, yellow-brown, circular or lentiform, 105.0-150.0 x 125.0-162.5 μm, with a circular pore, 20.0 x 22.5 μm in diam., surrounded by small dark cells. conidia cylindrical, both ends broadly rounded, straight, very rarely slightly bent, 1-septate, not constricted, 7.5-10.0 x 2.5-3.0 μm. morphological features of the specimens generally correspond to those listed by melnik (2000). in pycnidia there is also a contribution of smaller, unicellular conidia, 5.5-7.5 x 2.5-3.0 μm. material examined. on plantago intermedia gilib., central poland, łódź, piłsudskiego park, lawn, 24 june 2005, leg. e. połeć, lod pf 3135; south poland, wola near pszczyna, meadow of molinion caeruleae alliance, 11 oct. 2006, leg. a. myszka lod pf 2871. notes. the species has been reported on living leaves of plantaginaceae members: plantago aristata michx., p. asiatica l., p. depressa willd., p. major l. s. str., p. media l., p. rugelii decne. and plantago sp. as well as cyperaceae taxa: carex arenaria and carex sp. from asia (china, kazakhstan), north america (usa) and many european countries (melnik 2000; farr et al. 2011). in poland (fig. 4) it has been so far observed only on plantago major l. s. str. as ascochyta plantaginis in the słowiński national park (adamska 2001), in the pieniny mts (kućmierz 1976a, b, 1977), in szczecin city and its vicinity (madej 1974). it is reported on plantago intermedia gilib. for the first time. however, as p. intermedia was previously included in plantago major l. as subspecies intermedia (dc.) arcang., thus some of the records of ascochyta sodalis on p. major may, in fact, concern p. intermedia. interesting ascochyta species 197 ascochyta verbascina thüm. leaf spots oval or irregular, light brown or whitish. pycnidia scattered or aggregated in the small groups, immersed, pale yellow, brown to almost black, globosedepressed and lentiform, 92.5-107.5 x 102.5-172.5 μm, with a circular pore, 20.0-25.0 μm in diam., surrounded by small dark cells. conidia cylindrical, both ends rounded, straight or slightly bent, 0-1-septate, not constricted, 6.25-10.0 x 2.2-2.5 μm. morphological features of the specimen generally correspond to those described by melnik (2000). material examined. on veronica chamaedrys l. s. str., central poland, łódź, łagiewnicki forest, roadside, 06 oct. 2006, leg. e. połeć, lod pf 3137. notes. the species has been noted on living leaves and dry stems of scrophularia­ ceae members: rhinanthus minor l., verbascum blattaria l., v. densiflorum bertol., v. nigrum l., v. sinuatus l., verbascum sp., veronica beccabunga l., v. chamaedrys l. s. str., v. officinalis l. and v. urticifolia jacq., as well as on lamiaceae representative – scutellaria altissima l. from europe (austria, bulgaria, hungary, latvia, russia, ukraine) (melnik 2000; farr et al. 2011). in poland (fig. 5) it has so far been observed on verbascum densiflorum bertol. in the dendrological garden in przelewice (near the szczecin city) (madej 1969; sałata 2002), and on v. lychnitis l. and veronica sp. in olsztyn near częstochowa on the wyżyna częstochowska upland (połeć 2005; ruszkiewicz-michalska 2006). it is recorded on veronica chamaedrys l. s. str. for the first time in poland. final remarks although the genus ascochyta has been monographed in poland quite recently (sałata 2002), 60 new species for poland were reported within the following years (mułenko, kozłowska 2008; wołczańska 2010). the recent finding of eleven interesting species of this genus, including two new for poland, also proves that further investigations in natural and anthropogenic habitats are needed. the taxonomy of ascochyta species changes in time. according to punithalingam (1988) 1-septate conidia are the norm for the genus ascochyta. however, the occurrence of a small percentage of unicellular and 2-septate conidia is possible and it does not exclude the species from the genus. the subgenus ascochyta is characterized with the admixture of 3-4-celled conidia (melnik 2000). in the opinion of boerema et al. (2004) mature conidia of true ascochyta species are almost always septate and their conidiogenesis differs from the one observed in phoma species. the conidia of phoma taxa are always unicellular when released, but some of them can become twoor more-celled by secondary septation. according to boerema et al. (2004) many taxa referred to as ascochyta species are, in fact, the members of phoma genus. this concerns some of the species presented in the current report: ascochyta leonuri (= a. nepetae) is a synonym of phoma nepeticola (melnik) dorenb. & gruyter as well as ascochyta daturae and ascochyta sonchi are the synonyms of boeremia exigua var. exigua. in the checklist of 198 e. połeć and m. ruszkiewicz-michalska polish micromycetes these species are still numbered among the ascochyta species (mułenko, kozłowska 2008). to distinguish members of the genus ascochyta from phoma species with secondary conidial septation, the in vitro studies are needed (boerema et al. 2004). thus, the revision of the genus ascochyta based on in vitro studies and molecular analyses is necessary. our observations indicate that some of the species included currently to the genus ascochyta may also belong to phoma fr. s.l. (e.g., ascochyta infuscans and ascochyta sodalis), as in their pycnidia there is a contribution of unicellular conidia of variable shape and size. acknowledgements. the authors are indebted to professor maria ławrynowicz, curator of the fungal collection of herbarium universitatis lodziensis (lod), for permission to analyse herbarium materials. they include single specimens collected by msc students supervised by the second author: aleksandra kotynia, magdalena jakiel, danuta papierz and agnieszka myszka, whose contribution is greatly acknowledged. we are also grateful to two anonymous reviewers for valuable remarks on the manuscript. the studies were partially supported by the ministry of science and higher education (grant no n305 077 32/2708). the first author was also granted in the frame of the project “scholarships to support innovative doctoral research” by the european social fund and the budget as a part of integrated regional operational programme. references adamska i. 2001. microscopic fungus-like organisms and fungi of the słowiński national park. ii. (nw poland). acta mycol. 36 (1): 31–65. aveskamp m., de gruyter h., woudenberg j., verkley g., crous p.w. 2010. highlights of the didymel­ laceae: a polyphasic approach to characterise phoma and related pleosporalean genera. stud. mycol. 65: 1-64. boerema g. h., de gruyter j., noordeloos m. e., hamers m. e. c. 2004. phoma identification manual. differentiation of specific and infra-specific taxa in culture. cabi publishing, wallingford, 448 pp. buchanan p. k. 1987. a reappraisal of ascochytula and ascochytella (coelomycetes). mycol. pap. 156: 1–83. chlebicki a. 2002. biogeographic relationships between fungi and selected glacial relict plants. the use of host – fungus data as an aid to plant geography on the basis of material from europe, greenland and northern asia. monogr. bot. 90: 1–230. daisie: delivering alien invasive species inventories for europe; www.europe-aliens.org danilkiewicz m. 1990. parasitic fungi of river bug valley. acta mycol. 23 (2): 37–80. diedicke h. 1915. kryptogamenflora der mark branderburg. bd. ix. pilze vii. sphaeropsideae, melan­ conieae. verl. von gebrüder borntraeger, leipzig, 962 pp. durska b. 1974. studies on parasitic fungi of plants occuring in the lake littoral of the masurian lakeland. acta mycol. 10 (1): 73–139. faliński j. b., mułenko w. (eds). 1997. cryptogamous plants in the forest communities of białowieża national park. ecological atlas (project crypto 4). phytocoenosis 9 (n.s.) supplementum cartographiae geobotanicae 7: 1–522. farr d. f., rossman a.y., palm m.e. & mccray e.b. 2011. fungal databases, systematic botany & mycology laboratory, ars, usda [retrieved 17.05.2011, from the database at http://nt.ars-grin. gov/fungaldatabases/]. kirk p.m., cannon p.f., minter d.w., stalpers j.a. 2008. ainsworth & bisby’s dictionary of the fungi. 10th ed. cab international, wallingford: p. 54. kowalski t. 2004. endophytic fungi: vi. mycobiota in living symptomless leaves of ulmus glabra and in necrotic tissues associated with gall-making insects. phytopathol. pol. 32: 61-73. kozłowska m., mułenko w. 2005. notes on some species of ascochyta (coelomycetes) new and rare for poland. acta mycol. 40 (1): 43–47. kućmierz j. 1976a. new and rare for poland species of deuteromycetes collected in the area of the pieniny mts (western carpatians). fragm. flor. geobot. 22: 141–146. interesting ascochyta species 199 kućmierz j. 1976b. flora of parasitic fungi of the pieniny mountains (western carpatians). part ii. basidiomycetes, deuteromycetes. fragm. flor. geobot. 22: 605–622. kućmierz j. 1977. investigation on the parasitic fungi from the pieniny mts. zeszyty naukowe akademii rolniczej w krakowie 137, rozprawy 52: 1–142. madej t. 1969. mycoflora of herbs of the dendrologic garden in przelewice (szczecin voivode). fragm. flor. geobot. 15: 99–110. madej t. 1974. the materials for mycoflora of plants of szczecin province. akademia rolnicza w szcze-szczecinie, rozprawy 35: 1–235. melnik v. 2000. key to the fungi of the genus ascochyta lib. (coelomycetes). mitteilungen aus der biologischen bundesanstalt für landund forstwirtschaft berlin-dahlem. heft 379. berlin, 192 pp. miczyńska z. 1966. materials to the knowledge of diseases occuring on medical herbs cultivated in poland. i. diseases of medical herbs of the families: labiatae, scrophulariaceae and boraginaceae observed during the years 1951-1962. rocz. nauk roln., ser. a 92 (2): 285–312. mirek z., piękoś-mirkowa h., zając a., zając m. 2002. flowering plants and pteridophytes of poland. a checklist. (in:) z. mirek (ed.). biodiversity of poland 1. w. szafer institute of botany, polish academy of sciences, kraków, 442 pp. mułenko w. 1996. parasitic microfungi and their hosts collected on the study area. (in:) j. b. faliński, w. mułenko (eds). cryptogamous plants in the forest communities of białowieża national park. functional groups analysis and general synthesis (project crypto 3). phytocoenosis 8 (n.s.), archiv. geobot. 6: 55–65. mułenko w., kozłowska m. 2008. genus ascochyta. (in:) w. mułenko, t. majewski, m. ruszkiewiczmichalska (eds). a preliminary checklist of micromycetes in poland. (in:) z. mirek (ed.). biodiversity of poland 9. w. szafer institute of botany, polish academy of sciences, kraków: 514–529. mułenko w., kozłowska m., sałata b. 2004. microfungi of the tatra national park. a checklist. (in:) z. mirek, m. ronikier (eds). biodiversity of the tatra national park. 1. w. szafer institute of botany, polish academy of sciences, kraków, 72 pp. mułenko w., wojdyło b. 2002. mikroskopijne grzyby pasożytnicze drzew i krzewów arboretum bolestraszyce. arboretum bolestraszyce 9: 5–14. piątek m., wołczańska a. 2004. some phytopathogenic fungi rare or new to poland. polish bot. j. 49 (1): 67–72. połeć e. 2005. contribution to the knowledge of the phytopathogenic micromycetes of the częstochowa upland. (in:) k. czyżewska, j. hereźniak (eds). biodiversity in relation to vegetation zones in europe. university of łódź publishing house, łódź: 187–193. połeć e. 2010. grzyby pasożytnicze roślin terenu miasta łodzi. praca doktorska wykonana w katedrze algologii i mikologii uniwersytetu łódzkiego, 108 + lxxx pp. (mscr). punithalingam e. 1979. graminicolous ascochyta species. mycol. pap. 142: 1–214. punithalingam e. 1988. ascochyta ii. species on monocotyledons (excluding grasses), cryptogams and gymnosperms. mycol. pap. 159: 1–235. ruszkiewicz-michalska m. 2006. phytoparasitic micromycetes in plant communities of the wyżyna częstochowska upland. monogr. bot. 96: 1–140. rutkowski l. 2004. klucz do oznaczania roślin naczyniowych polski niżowej. pwn, warszawa, 814 pp. sałata b. 2002. polskie gatunki grzybów mitosporowych z rodzaju ascochyta. wyd. uniwersytetu marii curie-skłodowskiej, lublin, 121 pp. sałata b., mułenko w., wołczańska a. 1994. new and rare species of sphaeropsidales to the polish flora. acta mycol. 29 (1): 81–93. szafer w., kulczyński s., pawłowski b. 1986. rośliny polskie. pwn, warszawa, 1019 pp. wołczańska a. 2010. interesting collections of phytopathogenic fungi. acta mycol. 45 (1): 91–96. 200 e. połeć and m. ruszkiewicz-michalska interesujące gatunki grzybów z rodzaju ascochyta streszczenie w pracy przedstawiono jedenaście gatunków mikroskopowych grzybów pasożytniczych należących do rodzaju ascochyta (grzyby anamorficzne). dwa z nich – ascochyta bondarceviana melnik (na ribes sp.) i ascochyta equiseti (desm.) grove (na equisetum arvense l.) – są nowe dla bioty kraju, a pozostałe dziewięć gatunków stwierdzono na żywicielach, na których nie były one dotąd obserwowane w polsce. grzyby te zostały zebrane w polsce środkowej i południowej, przy czym większość z nich została zaobserwowana na terenie łodzi. dla wszystkich gatunków podano krótkie opisy cech struktur morfologicznych i rozmieszczenie, a gatunki notowane w polsce po raz pierwszy zostały również zilustrowane mikrofotografiami. 2014-01-01t23:48:40+0100 polish botanical society bibliography of publications by barbara gumińska władysław wojewoda1 and piotr mleczko2 1w. szafer institute of botany, polish academy of sciences, lubicz 46, pl-31-512 kraków 2institute of botany, jagiellonian university, lubicz 46, pl-31-512 kraków we present the complete list of publications by professor barbara gumińska, published in the period 1953-2009. it contains oryginal papers, monographs, scientific notes, books, book chapters and articles for the general public. the bibliography is arranged in chronological order. titles of papers written only in polish have been translated into english, and the publications are marked “in polish”. book reviews follow original and other papers in each year. 1953. nowe stanowisko trufli w pieninach. [new locality of truffle in pieniny mts.]. chrońmy przyrodę ojczystą 9 (3): 40 (as b. lubelska; in polish). 1954. o występowaniu trufli (tuber mich. i choiromyces vitt.) w polsce [the occurrence of the truffle (tuber mich. and choiromyces vitt.) in poland]. fragmenta floristica et geobotanica 1 (1): 87-95 (as b. lubelska; in polish with english summary). 1956. grzyby przyczyniające się do zamierania lasu osłabionego pożarem [fungi contributing to the die-back of a forest weakened by fire]. fragmenta floristica et geobotanica 2 (2): 112-133 (as b. lubelska-gumińska; in polish with english summary). 1957. repeated findings of the fungus wawelia regia namysł. in cracow. bulletin de l’académie polonaise des sciences cl. ii. 5 (10): 347-348. 1958. karel cejp, houby i. praha 1957. wiadomości botaniczne 2 (2): 87 (a review; in polish). 1959. phylloporus rhodoxanthus (schw.) bres. w polsce [phylloporus rhodoxanthus (schw.) bres. in poland]. fragmenta floristica et geobotanica 5 (1): 151154 (in polish with english summary). – wycieczka mikologiczna w bieszczady [mycological excursion to bieszcza-bieszczady mts.]. wiadomości botaniczne 1959 (2): 136 (in polish). – karel cejp, houby ii. praha 1958. wiadomości botaniczne 3 (4): 242-243. (a review; in polish). 1960. (domański s., gumińska b., lisiewska m., nespiak a., skirgiełło a., truszkowska w.) mikoflora bieszczadów zachodnich (wetlina 1958) [mycoflore acta mycologica vol. 45 (1): 11–16 2010 12 w. wojewoda and p. mleczko des bieszczady occidentales (wetlina 1958)]. monographiae botanicae 10 (2): 159-237 (in polish with french summary). – valentina and gordon r. wasson: mushrooms, russia and history. pantheon books, new york 1957. str. 433, rycin 28, tablic 82. wiadomości botaniczne 4 (2): 242-243 (a review; in polish). 1961. purchawka olbrzymia – największy grzyb na świecie [calvatia gigantea – the largest fungus of the world]. chrońmy przyrodę ojczystą 17 (5): 16-18 (in polish). – alina skirgiełło: grzyby (fungi) – podstawczaki (basidiomycetes) – borowikowe (boletales). flora polska, rośliny zarodnikowe polski i ziem ościennych. instytut botaniki polskiej akademii nauk, pwn, warszawa 1960. stron 130, tablic kolorowych 30, rycin kreskowych 47. wiadomości botaniczne 5 (3): 265 (a review; in polish). 1962. grzyby roztoki małej w beskidzie sądeckim [fungi of roztoka mała in the beskid sądecki mts.]. fragmenta floristica et geobotanica 8 (2): 205-213 (in polish with english summary). – mikoflora lasów bukowych rabsztyna i maciejowej (studium florystycznoekologiczne) [the fungi of the beech forests of rabsztyn and maciejowa (a study of floristic and ecological conditions). monographiae botanicae 13: 3-85 + 6 tables (in polish with english summary). 1963. (domański s., gumińska b., lisiewska m., nespiak a., skirgiełło a., truszkowska w.) mikoflora bieszczadów zachodnich . ii. (ustrzyki górne 1960) [mycoflora of west bieszczady. ii]. monographiae botanicae 15: 3-75 (in polish with english summary). – parki narodowe i rezerwaty jako tereny nowszych badań mikologicznych [national parks and nature reserves as territories of recent mycological investigations]. chrońmy przyrodę ojczystą 19 (3): 7-13 (in polish). 1965. grzyby rosnące pod ziemią i ich rola w przyrodzie [hypogeous fungi and their importnace in nature]. chrońmy przyrodę ojczystą 21 (6): 20-24 (in polish). 1966. mikoflora lasów jodłowych okolic muszyny [mycoflora of the fir forests of muszyna]. acta mycologica 2: 107-149 (in polish with english summary). – o kilku grzybach lasu jodłowego. wszechświat 6: 140-142 (in polish). – der pieniny national park. (in:) czwarty kongres europejskich mikologów. guide quatrième congrès des mycologues européens. warszawa, p. 99105. – (gumińska b., nespiak a.) auf der strecke zakopane-szczawnica-zakopane. (in:) czwarty kongres europejskich mikologów. guide quatrième congrès des mycologues européens. warszawa, p. 97-98. – lászlo szemere: die unterirdische pilze des karpatenbeckens (fungi hypogaei territorii carpato-pannonici). nakładem akadémiai kiadó, budapest 1965. stron 319, w tekście 8 czarno-białych rysunków, przy końcu książki 10 kolorowych tablic. wiadomości botaniczne 10 (4): 281-283 (a review; in polish). 1967. (domański s., gumińska b., lisiewska m., nespiak a., skirgiełło a., truszkowska w.) mikoflora bieszczadów zachodnich. iii (baligród 1962) [mycoflora of west bieszczady. iii]. acta mycologica 3: 63-114 (in polish). bibliography of publications by barbara gumińska 13 – o leśnych grzybach workowych [about forest ascomycetes]. wszechświat 7: 57-61 (in polish). 1968. sarcosphaera eximia (dur. et lév.) r. mre. w pienińskim parku narodowym [sarcosphaera eximia (dur. et lév.) r. mre. in the pieniny national park]. acta mycologica 4 (1): 131-146 (in polish with english summary). – (gumińska b., wojewoda w.) grzyby owocnikowe i ich oznaczanie [macrofungi and their identification]. pwril, warszawa, p. 1-308 (in polish). 1969. mikoflora pienińskiego parku narodowego (część i) [mycoflora of the pie-pieniny national park (part i)]. acta mycologica 5: 219-243 (in polish with english summary). 1970. rzadkie i nowe dla polski grzyby z rodziny hysterangiaceae znalezione w pienińskim parku narodowym [rare fungi of the family hysterangiaceae found in the pieniny national park]. fragmenta floristica et geobotanica 16 (3): 433-442 (in polish with english summary). 1972. nowe stanowisko ischnoderma corrugis w polsce [new locality of ischno­ derma corrugis in poland]. acta mycologica 8 (1): 141-143 (in polish with english summary). – mikoflora pienińskiego parku narodowego (część ii) [mycoflora of the pieniny national park (part ii)]. acta mycologica 8 (2): 149-174 (in polish with english summary). 1973. flora grzybów wielkoowocnikowych [flora of macrofungi]. (in:) przewodnik wycieczkowy 41 zjazdu ptb [excursion guide of 41st congress of the polish botanical society]. kraków, p. 17 (in polish). – i. turowska, z. podbielkowski, w. wojewoda: rośliny zarodnikowe. skrypt wydany przez akademię medyczną w krakowie, kraków 1971, tekst 195 stron, atlas 158 rycin na 51 stronach, cena skryptu + atlas 14 zł. wiadomości botaniczne 17 (1): 64-67 (a review; in polish). 1974. nowe stanowisko stropharia albocrenulata (peck.) kreisel znalezione w pie-pieninach [new locality of stropharia albocrenulata (peck.) kreisel in poland]. zeszyty naukowe uj, prace botaniczne 2: 201-208 (in polish with english summary). – z badań nad mikoflorą pienińskiego parku narodowego (wpływ warunków atmosferycznych na owocowanie grzybów) [investigations on mycoflora of pieniny national park (influence of weather conditions on fructification of fungi)]. (in:) a. skirgiełło, b. sałata (eds) materiały ogólnopolskiego sympozjum mikologicznego [materials of the polish mycological symposium]. lublin, p. 19-24 (in polish). 1975. a. n. wasiliewa: agarikowyje szlapocznyje griby primorskogo kraja. izd. „nauka”, akademia nauk sssr, leningrad 1973, str. 328, 67 rys., 14 tabel, 2 tablice barwne. wiadomości botaniczne 19 (1): 328-329 (a review; in polish). 1976. mikoflora pienińskiego parku narodowego (część iii) [mycoflora of the pieniny national park (part iii)]. zeszyty naukowe uj, prace botaniczne 4: 127-141 (in polish with english summary). – macromycetes łąk w pienińskim parku narodowym [macromycetes of meadows in pieniny national park]. acta mycologica 12 (1): 3-75 (in polish with english summary). 14 w. wojewoda and p. mleczko 1977. the locality of onygena equina (willd. ex s.f. gray) pers. ex fr. in the suburbs of cracow. zeszyty naukowe uj, prace botaniczne 5: 153-156. 1980. m. lisiewska, m. szmid: przewodnik grzyboznawczy, wyd. ii zmienione, pwril, warszawa 1980, 150 str., 74 ryc., 7 tabel. wiadomości botaniczne 24 (4): 293-294 (a review; in polish). 1981. mikoflora pienińskiego parku narodowego (część iv) [mycoflora of the pieniny national park (part iv)]. zeszyty naukowe uj, prace botaniczne 9: 67-81 (in polish). – w. rudnicka-jezierska: chaetomiales – czuprynkowe; b. sałata: proto­ mycetales – pierwogrzybowe. flora polska, rośliny zarodnikowe polski i ziem ościennych, grzyby (mycota), workowce (ascomycetes), tom 12, pwn, warszawa-kraków 1979, 216 str., 97 ryc., 10 fotografii (w 3 tablicach). wiadomości botaniczne 25 (3): 234-235 (a review; in polish). 1982. grzyby kapeluszowe pienińskiego parku narodowego [mushrooms of the pieniny national park]. (in:) k. zarzycki (ed.) przyroda pienin w obliczu zmian [nature of pieniny mts in result of changes]. pwn, warszawakraków, p. 189-209 (in polish). – a. skirgiełło, m. zadara: mucorales – pleśniakowe; m. ławrynowicz: endogonales – kłębiankowe, flora polska, rośliny zarodnikowe polski i ziem ościennych, grzyby (mycota), tom 10, pwn, warszawa-kraków 1979, 321 str., 97 ryc., 4 fotografie (w 2 tablicach). wiadomości botaniczne 26 (1/2): 70-71 (a review; in polish). – michael e., hennig b., kreisel h.: handbuch für pilzfreunde vierter band. blätterpilze – dunkelblätter. wydanie drugie. veb g. fischer verlag jena 1981, 472 str., 146 barwnych tablic (pojedynczych lub zbiorczych), 4 ryc., 16 fotografii czarno-białych i 7 mapek. wiadomości botaniczne 26 (3): 151 (a review; in polish). 1983. (gumińska b., wojewoda w.) grzyby i ich oznaczanie [fungi and their identification]. wydanie ii, zmienione. pwril, warszawa, p. 1-504 (in polish). – k. wells, e. k. wells: basidium and basidiocarp: evolution, cytology, function and development. springer-verlag, new york-heidelberg-berlin 1982, str. 187. wiadomości botaniczne 27 (1): 77-78 (a review; in polish). 1984. (gumińska b., turnau k.) repeated find of lycogala flavo-fuscum (ehrb.) rost. in cracow. zeszyty naukowe uj, prace botaniczne 12: 181-184. – j. webster: pilze. eine einführung. springer-verlag. berlin-heidelbergnew york 1983, str. 641, ryc. 332. wiadomości botaniczne 28 (1): 95-96 (a review; in polish). 1985. mutinus ravenelii (berk. et curt.) e. fisher (phallales, mycota) – nowy gatunek dla flory polski [mutinus ravenelii (berk. et curt.) e. fisher (phallales, mycota) – the species new to poland]. zeszyty naukowe uj, prace bota-botaniczne 13: 97-103 (in polish with english summary). – (gumińska b., wojewoda w.) grzyby i ich oznaczanie [fungi and their identification]. wydanie iii. pwril, warszawa, p. 1-305 (in polish). 1987. błąd w serii poświęconej grzybom [error in the series with fungi]. filatelista 34 (10): 237-238 (in polish). bibliography of publications by barbara gumińska 15 – odkrycie nowego gatunku grzyba z rodzaju wawelia [the discovery of a new species of the genus wawelia]. wiadomości botaniczne 31 (1): 9-13 (in polish with english summary). 1988. szybkość przyrastania „czarcich kręgów” na łące w okolicy muszyny [annual increase of „fairy rings” in a meadow in the neighbourhood of muszyna]. acta mycologica 24 (1): 51-58 (in polish with english summary). – szybkość przyrastania „czarciego kręgu” na łące w okolicy muszyny [growth rate of „fairy rings” in a meadow near muszyna]. folia societatis scientiarum lublinensis 30, biologia 1-2: 21-22 (in polish with english and russian summaries). – mikologia w filatelistyce [mycology in stumps-collecting]. wszechświat 8 (3): 65-67 (in polish). – (gumińska b., wojewoda w.) grzyby i ich oznaczanie [fungi and their identification]. wydanie iv. pwril, warszawa, p. 1-505 (in polish). – j. kochman: zarys mikologii dla fitopatologów. 2 wydanie, poprawione i uzupełnione. wydawnictwo sggw-ar warszawa 1986, 428 str., 234 ryc., 7 tabel, nakład 1000 egz. wiadomości botaniczne 32 (3): 186-187 (a review; in polish). – f. kotlaba: zemĕpi sné rozšiřeni a ekologie chorošci (polyporales s.1) ν československu. 194 str., 123 mapy, 86 fotografii w 36 tabelach. československa akademie vĕd. praha 1984, wydanie i. wiadomości botaniczne 32 (4): 250 (a review; in polish). 1989. macromycetes of the pieniny national park (a guide). (in:) 19th international phytogeographic excursion 1989, july 7-26, “flora and vegetation of poland – changes, management and conservation: 1928–1988”. institute of botany, polish academy of sciences, kraków, p. 1-34. – m. semerdžieva, j. veselský: lečivé houby dřive a nyni. praha 1986. wydanie i. 177 str., 28 ryc., 2 tabele, 32 fotografie barwne. československé akademie věd. wiadomości botaniczne 33 (1): 44-46 (a review; in polish). 1990. mikoflora pienińskiego parku narodowego (część v) [mycoflora of the pieniny national park (part v)]. zeszyty naukowe uj, prace botaniczne 21: 157-172 (in polish with english summary). 1991. (gumińska b., heinrich z., olech m.) macromycetes of the nw sørkapp land, spitsbergen. polish polar research 12 (3): 407-417. – (gumińska b., marecka h.) cis taxus baccata l. w rezerwacie „cisy w malinówce” (województwo krośnieńskie) [yew tree taxus baccata l. in the „cisy w malinówce” nature reserve (district krosno)]. ochrona przyrody 48: 105-119 (in polish with english summary). 1992. (gumińska b., mierzeńska m.) gerronema marchantiae sing. et clem. – a fungus associating with marchantia polymorpha l. and nostoc sp. zeszyty naukowe uj, prace botaniczne 24: 171-177. – higher fungi of the tilio­carpinetum forest association in the skołczanka reserve near cracow. acta mycologica 27 (1): 137-158. – macromycetes of the pieniny national park (s. poland). (in:) k. zarzycki, e. landolt, j.j. wójcicki (eds) contributions to the knowledge of flora and vegetation of poland. proceedings of the 19th international phytogeographic excursion (ipe), 1989, through sounthern poland. volume 2. flora 16 w. wojewoda and p. mleczko and vegetation of the regions visited during the 19th ipe. present and former state. veröffentlichungen des geobotanischen institutes eth, stiftung rübel, zürich, 107: 238-252. – grzyby wybranych łąk w pienińskim parku narodowym – stan aktualny i warunki jego zachowania [macromycetes of some meadows in the pieniny national park – the present state and suggestions for the future management]. pieniny – przyroda i człowiek 2: 65-70 (in polish with english summary). – kilka uwag o encyklopedii guinessa [some notes on the guiness encyclopedia]. wszechświat 93 (3): 84-85 (in polish). 1994. mikoflora pienińskiego parku narodowego (część vi) [mycoflora of the pieniny national park (part vi)]. fragmenta floristica et geobotanica series polonica 1: 33-39 (in polish with english summary). – (gumińska b., heinrich z., olech m.) macromycetes of the south shetland islands (antarctica). polish polar research 15 (3-4): 103-109. 1997. flora polski. grzyby (mycota) 26: podstawczaki (basidiomycetes), wodnichowate (hygrophoraceae), uniwersytet jagielloński, instytut botaniki, kraków, pp. 202, 22 pls. (in polish with english key). – (dyląg e., gumińska b.) postfire macromycetes from deciduous wood in the chrzanów forest inspectorate (s poland). acta mycologica 32 (2): 173-187. – (gumińska b., turnau k.) okazały owocnik grzyba podziemnego choiromy­ ces meandriformis vitt. z beskidu wyspowego [impressive fruitbody of the hypogeous fungus choiromyces meandriformis vitt. from beskid wyspowy mts.]. chrońmy przyrodę ojczystą 53 (2): 90-93 (in polish). 1999. mikoflora pienińskiego parku narodowego (część vii) [mycoflora of the pieniny national park (part vii)]. fragmenta floristica et geobotanica series polonica 6: 179-187 (in polish with english summary). 2000. grzyby wielkoowocnikowe (macromycetes). (in:) j. razowski (ed.) flora i fauna pienin – monografie pienińskie 1: 47-53 (in polish). – resupinatus kavinii (tricholomataceae), a fungus species newly noted in poland. fragmenta floristica et geobotanica 45 (1-2): 509-512. – wawelia – historia i nowe odkrycia [wawelia – the history and current news]. wiadomości botaniczne 44 (3-4): 23-25 (in polish with english summary). 2002. new localities of two interesting species of fungi growing on sawdust. polish botanical journal 47 (2): 139-142. 2004. (gumińska b., wojewoda w.) mikoflora pienińskiego parku narodowego (część viii) [mycoflora of the pieniny national park (part viii)]. fragmenta floristica et geobotanica series polonica 11 (2): 371-382 (in polish with english summary). 2006. atlas grzybów pienińskiego parku narodowego [atlas of fungi of pieniny national park]. wyd. pieniński park narodowy, krościenko n/dunajcem, pp. 166 (in polish). 2009. aksamitka złota pholiota aurea: grzyb rzadki w polsce [pholiota aurea: a rare fungus in poland]. chrońmy przyrodę ojczystą 65 (3): 205-208 (in polish with english summary). 2014-01-01t11:50:04+0100 polish botanical society laboulbeniales (ascomycetes) from latvia andré de kesel and inguna krastina de kesel national botanic garden of belgium, domein van bouchout b 1860 meise, adk@br.fgov.be d e k e s e l a., k r a s t i n a d e k e s e l i.: laboulbeniales (ascomycetes) from latvia. acta mycol. 41 (1): 55 64, 2006. this contribution presents new and historical data on the laboulbeniales of latvian coleoptera. an annotated checklist of 26 taxa is given, 13 are new for latvia. only six taxa (accepted names) from briedis’ historical list were found again and six more need to be confirmed as briedis’ material was lost. a neotype is indicated here for the extremely rare laboulbenia elaphricola siemaszko et siemaszko. its morphology is discussed and compared with laboulbenia elaphri spegazzini and laboulbenia vulgaris peyr. key words: laboulbeniales, latvia, coleoptera, laboulbenia elaphricola introduction laboulbeniales (ascomycetes) are strictly parasitic on arthropoda, mainly insects. although widespread, information concerning their distribution in europe is unequal and, as yet, a reflection of the distribution of laboulbeniologists in europe (we i r , r o s s i 1995). the most exhaustive studies were carried out in spain and poland, followed by france, finland, britain, italy, belgium, germany and hungary. from other european countries, including the baltic countries, only very few and isolated reports or collections are available. records from estonia and lithuania have been published in s i e m a s z k o and s i e m a s z k o (1928, 1932), h u l d é n (1985) and m a r k o v s k a j a (2000) respectively. in latvia 16 species of laboulbeniales have been recorded in the literature (b r i e d i s 1932; h u l d é n 1985). all species were found on ground beetles (coleoptera, carabidae) and all, but one misgomyces, belong to laboulbenia. a thorough study of briedis’ material and notes was not possible as it could not be located; we consider it lost. this paper reports on newly collected material and literature records of laboulbeniales found on latvian coleoptera (carabidae and staphylinidae). acta mycologica vol. 41 (1): 55-64 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 56 a. de kesel and i. krastina de kesel materials and methods carabidae and staphylinidae (coleoptera) were captured by hand or by means of pitfall traps. sampling sites are situated in latvia and mainly include river associated forests, riverine marshes and banks along the river gauja and abuls (valmiera raijons). samples were also taken in the plant debris zones along the shorelines of the baltic sea, near jurmala (majori) and pabazi (balta kapa). hosts were killed and stored in 70 or 90% denaturated ethanol. screening hosts for infections and preparing of thalli was done using a stereomicroscope at high magnification (25-50x). the thalli were mounted and stained in permanent slides using a medium based on arabic gum with cotton blue and a trace of glycerine (d e k e s e l 1998). field data (locality, gps-coordinates, date and habitat), host data (taxon, gender and infection site) and parasite data (taxon, number of specimens and development) were recorded. slide(s) from the laboulbeniales and the corresponding hosts were given the same number (i.e. dkk-number); all material and notes are deposited at br (herbarium national botanic garden of belgium). when specific identity of a host was doubtful only the generic name of the host was used, followed by sp. the generic taxonomy of staphylinidae follows l o h s e (1964) and l o h s e et al. (1974), for carabidae l i n d r o t h (1974) or f r e u d e (1976). nomenclature and identification of all laboulbeniales found in latvia is largely based on ‘the laboulbeniales of poland’ by t. m a j e w s k i (1994). we consider it a key reference work for the study of the laboulbeniales from the baltics. to avoid redundancy we refrained from giving exhaustive species descriptions and comments, unless complementary to the information given in m a j e w s k i (l.c.) or elsewhere. results annotated and preliminary checklist of the laboulbeniales from latvia in the following checklist we placed species in bold when they correspond with verified records, i.e. species found by us in 2004-2005 and with reference material in herbarium br. species marked with an * are new for latvia. species in normal case have been mentioned in b r i e d i s (1932); their presence in latvia awaits confirmation. aphanandromyces audisioi w. rossi* • on brachypterus urticae (fabricius) (coleoptera, nitidulidae). dkk5-6: valmiera, gauja, tributary misa, 11.08.2004, n57°31.66’ – e025°26.47’, on flowers of urtica. thalli were found on the elytra and the abdominal tergites of the hosts. euzodiomyces lathrobii thaxt.* • on lathrobium sp. (coleoptera, staphylinidae). dkk17: valmiera, gauja, tributary misa, 12.08.2004, n57°31.00’ – e025°26.50’, under litter and freshwater algae along rivulet banks. thalli occur on the edges of the elytra. haplomyces texanus thaxt.* • on bledius sp. (coleoptera, staphylinidae). dkk9: valmiera, gauja, 05.08.2004, n57°31.66’ – e025°26.47’, riverside. thalli were found on the last abdominal tergite. laboulbenia argutoris cépède, picard* • on pterostichus diligens (sturm) (coleoptera, carabidae). dkk48: pabazi, laboulbeniales (ascomycetes) from latvia 57 fig. 1. a. laboulbenia elaphri spegazzini from pronotum of male elaphrus cupreus (dkk43); b e. laboulbenia elaphricola siemaszko et siemaszko, with b,d & e. from pronotum of male elaphrus riparius (dkk62) and c. juvenile thallus with typical stout habit, taken from right elytron of female elaphrus riparius (dkk38b); f. laboulbenia vulgaris peyr. from elytron of female bembidion sp. (dkk44). scale bar = 50μm. 58 a. de kesel and i. krastina de kesel balta kapa, 04.06.2005, n57°14’07.8” – e024°23’28.7”, in debris along beach. thalli were found on the elytra. • on pterostichus sp. (coleoptera, carabidae). dkk40: brenguli, river abuls, 31.05.2005, n57°32’56.2” – e025°32’24.3”, along rivulet bank in forested area. thalli were found on the elytra. laboulbenia calathi majewski • on calathus melanocephalus l., in zilupe (b r i e d i s 1932) laboulbenia clivinalis thaxt. • on clivina fossor l. (coleoptera, carabidae), in valka, latgale and zilupe (b r i e d i s 1932) • on clivina fossor l. dkk23: jurmala, majori, 02.06.2005, n56°58’ – e023°45’, in plant debris along the beach; dkk26: valmiera, gauja, 03.06.2005, n57°32’21.6” – e025°26’44.3”, river bank with pioneer vegetation of willows; dkk46: pabazi, balta kapa, 04.06.2005, n57°14’07.8” – e024°23’28.7”, in debris along beach. thalli are commonly found on both elytra and the metathorax. laboulbenia cristata thaxt.* • on paederus cf. riparius (coleoptera, staphylinidae). dkk45: pabazi, balta kapa, 04.06.2005, n57°14’07.8” – e024°23’28.7”, in debris along beach. thalli were found on the abdominal tergites, well protected by the elytra. laboulbenia dubia thaxt. • on philonthus fuscipennis mannh. and philonthus sp., (coleoptera, staphylinidae), in zilupe (b r i e d i s 1932). laboulbenia elaphri spegazzini* • on elaphrus cupreus dufts. (coleoptera, carabidae). dkk25(a,b,c,d), dkk28: valmiera, gauja, 03.06.2005, n57°32’21.6” – e025°26’44.3”, in river bank with pioneer vegetation of willows; dkk43: brenguli, river abuls, 31.05.2005, n57°32’56.2” – e025°32’24.3”, along rivulet bank. thalli were found on the right elytron, the pronotum and the mandibula. thalli from the latter spot have a smaller lower receptaculum and exhibit a more stout habitus. laboulbenia elaphricola siemaszko et siemaszko * • on elaphrus riparius (l.) (coleoptera, carabidae). dkk38(a,b): valmiera, gauja, tributary misa, 04.06.2005, n57°31.08’ – e025°26.52’, under litter, along rivulet bank in forested zone; dkk61, dkk62, dkk66: ibidem s.l., 03.06.2005, n57°32”0.5” – e025°26’49.2”, along rivulet banks; dkk42: brenguli, river abuls, 31.05.2005, n57°32’56.2” – e025°32’24.3”, along rivulet bank in forested area. thalli are found all over the hosts’ integuments, but most thalli occur on the elytra and the pronotum. laboulbenia elaphricola was described by s i e m a s z k o and s i e m a s z k o in 1928 (pol. pismo entomol. 6, pg. 200, t.vii, fig. 2). it was found in eastern poland, pulawy, on elaphrus riparius l. the original description is very short and the drawing of the type represents a damaged and incomplete outer appendage. the type material was unfortunately lost during wwii (m a j e w s k i 1994). since its description only b à n h e g y i (1950) reported l. elaphricola, but stated later that his material belongs to l. elaphri (b à n h e g y i 1964). l. elaphricola seems to be a very rare species, because very intensive screening of suitable habitats in eastern poland, bialowieza (m a j e w s k i 2003) did not supply new material. today it is a species with an uncertain status. in the absence of information on its full morphology and eventual posi laboulbeniales (ascomycetes) from latvia 59 tion related variations, b à n h e g y i (1964) and m a j e w s k i (1994) suggested that it could just be a growth form of the more common laboulbenia elaphri spegazzini. the numerous specimens we could collect on elaphrus riparius from latvia correspond with l. elaphricola. our material confirms that this species is not a growth form of l. elaphri as its typical short and stout thalli occur invariably on different parts of the hosts’ integument. l. elaphricola is indeed a small and dark species and no forms, even closely resembling l. elaphri, were seen on the studied hosts. we consider s i e m a s z k o and s i e m a s z k o ’ s l. elaphricola a good species. it is very distinct from l. elaphri and actually close to l. vulgaris, as already stated by s i e m a s z k o and s i e m a s z k o (1928). the following diagnosis and figure 1 should help to separate it from l. vulgaris. all thalli of l. elaphricola are small and stout, invariably of their origin on the host. they are never longer than 220μm (foot-ostiolum). already in an early stage of development l. elaphricola shows a deep pigmentation of the receptaculum. except for the poorly and uniformly pigmented cell i, cell ii and septum i-ii, all adult specimens have a deeply pigmented to blackish thallus. in l. vulgaris the septum i-ii is often darker, a feature that lacks in l. elaphricola. the perithecium is more free in l. elaphricola than in l. vulgaris, i.e. the insertion cell and cell v always being slightly below the middle of the perithecium in l. elaphricola. cell ii of l. elaphricola almost immediately widens upwards, forming a strikingly long septum with cell vi and a short septum with cell iii. cell vi is much broader than high. in most cases the septum ii-vi is at least twice the length of septum ii-iii. the upper receptacle, i.e. the complex of cells iii, iv and v, is delimited or demarcated from the rest of the thallus by a very deeply pigmented line, even in very young thalli. this darkened line starts at the anterior side of the insertion cell (close to the perithecium). it continues downwards along the adaxial septa of cell v, cell iv and cell iii, makes a curves along the basal septum of the latter, to finally end at the posterior (dorsal) side of the receptacle. the outer appendage is not branched and relatively long. in many cases, but not always, the three basal cells of the outer appendage tend to be slightly inflated. specimens with a branched outer appendage occur very rarely and only as a result of damage and subsequent atypical regeneration of the appendage. the inner appendage is branched once or twice and never exceeding the ostiolum. the insertion cell is constricted, sometimes strongly. the name bearing type material of l. elaphricola is lost and since its description l. elaphricola was not found again in poland or elsewhere (majewski pers. comm.). in this context we indicate our collection dkk62 (kept at br) from latvia as neotype for laboulbenia elaphricola siemaszko et siemaszko. this material was obtained from the elytra of elaphrus riparius l., caught on 03.vi.2005 along the forested banks of a tributary of the river gauja in valmiera (latvia, n57°31.08’ – e025°26.52’). laboulbenia fasciculata peyr. • on patrobus excavatus payk. (coleoptera, carabidae), in ogre, valka and valmiera (b r i e d i s 1932) • on patrobus atrorufus (stroem), dkk3 & 4: valmiera, gauja, tributary misa, 11.08.2004, n57°31.66’ – e025°26.47’, under debris along rivulet; dkk18, 19, 21: ibi60 a. de kesel and i. krastina de kesel dem, 12.08.2004. thalli occur on all parts of the integument, some hosts are heavily infected, carrying over 200 thalli. laboulbenia flagellata peyr. • on agonum albipes fabr. (as platynus ruficornis goeze), (coleoptera, carabidae), in koknese and ogre (b r i e d i s 1932) • on agonum assimile (paykull) (coleoptera, carabidae). dkk41: brenguli, river abuls, 31.05.2005, n57°32’56.2” – e025°32’24.3”, along rivulet bank in forested area; dkk52(a,b), dkk63: valmiera, gauja, tributary misa, 03.06.2005, n57°32”0.5” – e025°26’49.2”, along rivulet banks. infections occur on the upper part of the exoskeleton, i.e. the pronotum and both elytra. • on agonum viduum (panzer) (coleoptera, carabidae). dkk68: valmiera, gauja, tributary misa, 10.08.2005, n57°32”0.5” – e025°26’49.2”, in plant debris. thalli were found on the elytra. • on agonum sp. (coleoptera, carabidae). dkk22: valmiera, gauja, tributary misa, 10.08.2004, n57°31.00’ – e025°26.50’, under litter along rivulet banks. thalli occur on all parts of the integument. • on bembidion adustum schaum., b. ustulatum l. and bembidion sp., in valka (b r i e d i s 1932) • on bembidion fluviatile l., in valmiera (b r i e d i s 1932) remark: b r i e d i s (1932) reports mixed infection with l. luxurians, l. pedicellata and l. vulgaris. the presence of laboulbenia flagellata on bembidion is however doubtful and hitherto not mentioned on any species of bembidion in europe (s a n t a m a r í a et al. 1991; m a j e w s k i 1994). laboulbenia filifera thaxt. • on harpalus aeneus fabr. (coleoptera, carabidae), in valka (b r i e d i s 1932) laboulbenia hyalopoda de kesel * • on dromius linearis (olivier) (coleoptera, carabidae). dkk69: valmiera, gauja, tributary misa, 03.06.2005, n57°32’0.5” – e025°26’49”, riverine marsh, in dead stem of heracleum cf. sphondylium l. (hogweed). despite their length, thalli are very easily overlooked. they were found on the caudal extremity (soft parts) of the last abdominal segment. thallus very slender, hyaline to yellowish, up to 390 μm long. receptaculum slender, 210 μm high. cell i and ii three to four times higher than broad, without pigmentation. base of cell i not pigmented; foot hyaline, with one or two small brownish spots. cell iii and iv about the same height (20-25 μm), two times higher than broad. adaxial side of cell v free from the perithecium. cell v triangular, slightly rounded, half as high as cell iv. insertion cell amber to dark brown, moderately constricted. abaxial (outer) appendage straight, not branched, up to 80 μm long, probably longer in undamaged specimens, not pigmented and without dark septa; parafysopodium (basal cell of outer appendage) similar to other cells, up to 20 μm high. adaxial (inner) appendage hardly pigmented, up to 80μm long; andropodium up to 8μm long, bearing two simple appendages. antheridia not seen. cell vi smaller than cell iii, easily distinguished as is the rest of the perithecial basal cells. perithecium long, slender and slightly asymmetrically curved, 180 × 32 μm, widest below the middle, hyaline, without or almost without pigmentation at the abaxial side; perithecial apex is differentiated in a 70 μm long and slender abaxially bent laboulbeniales (ascomycetes) from latvia 61 neck. ostiolum hyaline, consisting of 4 strongly inflated cells (lips), the most adaxial cell (lip) up to 20μm shorter than the others. laboulbenia luxurians thaxt. • on bembidion adustum schaum, (coleoptera, carabidae), in sigulda (briedis 1932) • on bembidion dentellum (thunb.) (coleoptera, carabidae). dkk53 (mixed with l. vulgaris): valmiera, gauja, tributary misa, 03.06.2005, n57°32”0.5” – e025°26’49.2”, along rivulet banks. thalli occur on the abdomen. • on bembidion littorale oliv. (coleoptera, carabidae), in ogre (b r i e d i s 1932) • on bembidion tetracolum say. (coleoptera, carabidae). dkk54, dkk55(a,b), dkk58(a), dkk59, dkk65 (mixed with l. vulgaris): valmiera, gauja, tributary misa, 03.06.2005, n57°32”0.5” – e025°26’49.2”, along rivulet banks. thalli were found on all body parts, often on the pronotum, the elytra, the prothorax and the legs. • on bembidion ustulatum duft., b. andreae er., in sigulda (b r i e d i s 1932) remarks: b r i e d i s (1932) reports mixed infection with l. flagellata, l. pedicellata and l. vulgaris. in our material mixed infections were only observed on bembidion dentellum and bembidion tetracolum, i.e. mixed with l. vulgaris (see dkk53 and dkk58(a,b) & dkk65 respectively). laboulbenia pedicellata thaxt. • on dyschirius globosus herbst (coleoptera, carabidae), in zilupe (b r i e d i s 1932) • on dyschirius thoracicus (rossi) (coleoptera, carabidae), in riga and sigulda (h u l d é n 1985) • on dyschirius sp. (coleoptera, carabidae). dkk10, dkk13: valmiera, gauja, 05.08.2004, n57°31.66’ – e025°26.47’, along de riverside; dkk30(b), dkk31, dkk36, dkk37(a,b): valmiera, gauja, 03.06.2005, n57°32’22.1” – e025°26’41.3”, riverside, on sand ; dkk50 : pabazi, balta kapa, 04.06.2005, n57°14’07.8” – e024°23’28.7”, in debris along beach. thalli occur on the elytra, the proand metathorax and tibia. • on bembidion adustum schaum (coleoptera, carabidae), in valka (b r i e d i s 1932) • on bembidion sp. (coleoptera, carabidae), dkk11,12: valmiera, gauja, 05.08.2004, n57°31.66’ – e025°26.47’, along de riverside ; dkk24: jurmala, majori, 02.06.2005, n56°58’ – e023°45’, in plant debris along the beach; dkk51: pabazi, balta kapa, 04.06.2005, n57°14’07.8” – e024°23’28.7”, in debris along beach. thalli occur on the legs (femur) and the elytra. laboulbenia philonthi thaxt.* • on philonthus sp. (coleoptera, staphylinidae). dkk8: valmiera, gauja, 05.08.2004, n57°31.6’ – e025°26.5’, along the shoreline; dkk27: valmiera, gauja, 03.06.2005, n57°32’21.6” – e025°26’44.3”, in river bank with pioneer vegetation of willows.; dkk29: valmiera, gauja, 03.06.2005, n57°32’22.1” – e025°26’41.3”, riverside, on sand. thalli were found on the abdominal tergites and the cephalon. laboulbenia pseudomasei thaxt. * • on pterostichus nigrita (payk.) (coleoptera, carabidae). dkk47 : pabazi, balta kapa, 04.06.2005, n57°14’07.8” – e024°23’28.7”, in debris along beach; dkk60, 62 a. de kesel and i. krastina de kesel dkk64: valmiera, gauja, tributary misa, 03.06.2005, n57°32”0.5” – e025°26’49.2”, along rivulet banks. thalli were found on the elytra, the proand metathorax. proliferation of appendages is very common in adult thalli. • on pterostichus sp., dkk1-2: valmiera, gauja, tributary misa, 11.08.2004, n57°31.66’ – e025°26.47’, under debris along rivulet; dkk20: ibidem, 12.08.2004. thalli were found on various parts of the integument, i.e. legs, pronotum and both elytra. laboulbenia rigida thaxt. • on pterostichus nigrita (payk.) (coleoptera, carabidae), in valka (b r i e d i s 1932) laboulbenia rougetii montagne , robin • on chlaenius vestitus payk. (coleoptera, carabidae), near ogre (b r i e d i s 1932) laboulbenia thaxteri cépède, picard • on asaphidion flavipes (l.) (coleoptera, carabidae), in sigulda (h u l d é n 1985) laboulbenia vulgaris peyr. • on bembidion andreae er. (coleoptera, carabidae), in valmiera (b r i e d i s 1932) • on bembidion dentellum (thunb.). dkk53 (mixed with l. luxurians): valmiera, gauja, tributary misa, 03.06.2005, n57°32”0.5” – e025°26’49.2”, along rivulet banks. thalli occurred on the abdomen. • on bembidion lunatum duft. (coleoptera, carabidae), in sigulda (b r i e d i s 1932) • on bembidion saxatile gyll. (coleoptera, carabidae), in valka (b r i e d i s 1932) • on bembidion tetracolum say. (coleoptera, carabidae), dkk56, dkk57, dkk58(b), dkk65 (mixed with l. luxurians): valmiera, gauja, tributary misa, 03.06.2005, n57°32”0.5” – e025°26’49.2”, along rivulet banks. thalli were observed on the pronotum, the elytra and to a lesser degree also on the legs. • on bembidion sp. (coleoptera, carabidae). dkk39: valmiera, gauja, tributary misa, 04.06.2005, n57°31.08’ – e025°26.52’, under litter, along rivulet bank in forested zone; dkk44: brenguli, river abuls, 31.05.2005, n57°32’56.2” – e025°32’24.3”, along rivulet bank in forested area. hosts are infected on the elytra. remark: mixed infections with laboulbenia luxurians are often observed on bembidion tetracolum (cf. dkk65). misgomyces dyschirii thaxt. • on dyschirius globosus (herbst.) (coleoptera, carabidae): in sigulda and jugla (h u l d é n 1985) • on dyschirius sp. dkk30(a): valmiera, gauja, 03.06.2005, n57°32’22.1” – e025°26’41.3”, riverside, on sand; dkk35: ibidem s.l., n57°32’21.6” – e025°26’44.3”, sandy shores of gauja. thalli were found in the side edges from the elytra. monoicomyces brittanicus thaxt. * • on atheta sp. (coleoptera, staphylinidae, aleocharinae). dkk49(a,b,c): pabazi, balta kapa, 04.06.2005, n57°14’07.8” – e024°23’28.7”, in debris along the beach. an easily overlooked species. thalli were growing on the cephalon, between the antennae. laboulbeniales (ascomycetes) from latvia 63 peyritschiella protea thaxt.* • on anotylus sp. (coleoptera, staphylinidae). dkk32(a,b), dkk33: valmiera, gauja, 03.06.2005, n57°32’21.6” – e025°26’44.3”, river bank with pioneer vegetation of salix sp. thalli occur on the antennae, the legs and metasternum. rhadinomyces cristatus thaxt.* • on lathrobium sp. (coleoptera, staphylinidae). dkk15, 16: valmiera, gauja, tributary misa, 12.08.2004, n57°31.00’ – e025°26.50’, under litter and freshwater algae along rivulet banks. thalli occur on the metasternum, abdominal tergites and some of the coxae. synonyms and doubtful or excluded records mentioned in briedis (1932) the species listed below represent historical records from b r i e d i s (1932). all these taxa were either reduced into synonymy (s a n t a m a r i a et al. 1991 or m a j e w s k i 1994) or should be considered as doubtful records in the studied area. 1. laboulbenia brachiata thaxt. l. fasciculata peyr. 2. laboulbenia elongata thaxt. l. flagellata peyr. 3. laboulbenia europaea thaxt. l. rougetii montagne, robin 4. laboulbenia fumosa thaxt. • on agonum piceum l., in valka 5. laboulbenia harpali thaxt. • on harpalus pubescens müll., in zilupe • on harpalus tardus panz., h. hirtipes panz., h. aeneus fabr., in valka 6. laboulbenia parvula thaxt. doubtful in europe (s a n t a m a r í a et al. 1991) • on agonum assimile (as platynus assimilis payk.), near valka 7. laboulbenia polyphaga thaxt., i.e. var. calathicola probably to be considered a record of l. calathi conclusions twenty six species are reported for latvia, 13 are new for the studied territory. six species reported by b r i e d i s (1932) were confirmed. the presence of six more species, reported by b r i e d i s (1932) and h u l d é n (1985), needs to be confirmed with new material as briedis’ original material from latvia is untraceable. laboulbenia elaphricola siemaszko et siemaszko is reported for latvia. this taxon is extremely rare and not reported, with certainty, since its discovery. it has an uncertain status because of a very short original diagnosis. the name bearing type was unfortunately lost and l. elaphricola is currently considered a growth form of l. elaphri. the latvian material, however, provides enough evidence that l. elaphricola is a good species. 64 a. de kesel and i. krastina de kesel references b á n h e g y i j. 1950. ritka laboulbeniák a kárpátmedencéböl (laboulbeniales rares du bassin carpatique). ann. biol. univ. budapest. (budapesti tudományegyetem biol. intézeteinek évkönyve) 1: 189 196. b á n h e g y i j. 1964. notes sur quelques laboulbéniacées de la pologne. ann. univ. sci. budapest. rolando eötvös, sect. biol. 7: 19 27. b r i e d i s a. 1932. laboulbeniaceae in latvia (preliminary note). acta horti botanici universitatis latvien sis 7: 131 134. d e k e s e l a. 1998. identificatie en gastheerspectrum van het genus laboulbenia in belgië (ascomycetes, laboulbeniales). sterbeeckia 18:13 31. f r e u d e h. 1976. familie carabidae (laufkäfer). (in:) h. f r e u d e , k. w. h a r d e , g. a. l o h s e (eds). die käfer mitteleuropas. 2. adephaga 1. goecke , evers verlag, krefeld, germany. 302pp. h u l d é n l. 1985. floristic notes on palaearctic laboulbeniales (ascomycetes). karstenia 25: 1 16. l i n d r o t h c.h. 1974. handbooks for the identification of british insects. vol. iv, part 2: coleoptera, cara bidae. royal entomological society of london. united kingdom. 148pp. l o h s e g. 1964. staphylinidae i (micropeplinae bis tachyporinae). (in:) freude h., harde k.w., lohse g.a. die käfer mitteleuropas iv. goecke , evers, krefeld, germany. 264 pp. l o h s e g., b e n i c k g., l i k o w s k i z. 1974. staphylinidae ii (hypocyphtinae und aleocharinae). pselap hidae. (in:) f r e u d e h., h a r d e k.w., l o h s e g.a. die käfer mitteleuropas v. goecke , evers, krefeld, germany. 381 pp. m a j e w s k i t. 1994. the laboulbeniales of poland. polish bot. stud. 7: 3 466. m a j e w s k i t. 2003. distribution and ecology of laboulbeniales (fungi, ascomycetes) in the bialowieza for est and its western foreland. phytocoenosis vol. 15 (n.s.) 2003. supplementum cartographiae geobo tanicae 16 warszawa bialowieża. 144 pp. m a r k o v s k a j a s. 2000. data on lithuanian laboulbeniales. botanica lithuanica 6 (3): 299 311. s a n t a m a r i a s., b a l a z u c j., ta v a r e s i.i. 1991. distribution of the european laboulbeniales (fungi, ascomycotina). an annotated list of species. treballs de l’institut botánic de barcelona 14: 1 123. s i e m a s z k o j., s i e m a s z k o w. 1928. owadorosty polskie i palearktyczne. (laboulbeniales polonici et palaearctici.) polskie pismo entomol. 6: 188 211. tav. vii. s i e m a s z k o j., s i e m a s z k o w. 1932. owadorosty polskie i palearktyczne. (laboulbeniales polonici et palaearctici.). ii. polskie pismo entomol. 10: 149 188. tab. vii x. we i r a., r o s s i w. 1995. laboulbeniales parasitic on british diptera. mycol. res. 99 (7): 841 849. gatunki laboulbeniales (ascomycetes) z terenu łotwy s t r e s z c z e n i e w artykule przedstawiono zarówno nowe jak i historyczne dane o gatunkach laboulbe niales występujących na łotewskich coleoptera, przede wszystkim na carabidae i staphyli nidae. odnotowano 26 gatunków, z których 13 jest nowych dla łotwy. pierwsza lista gatun ków laboulbeniales z łotwy została opublikowana w roku 1932 przez briedisa w acta horti botanici universitatis latviensis. kolekcja briedisa najprawdopodobniej zaginęła. sześć po twierdzonych taksonów z listy briedisa zostało na nowo odnalezionych, a dalsze sześć należy odszukać. krytyczna lista 26 gatunków zawiera również uwagi taksonomiczne dotyczące m. in. laboulbenia elaphricola siemaszko et siemaszko. 2014-01-01t11:43:29+0100 polish botanical society characterization of phoma negriana thüm., a new species from grapevine canes zofia machowicz-stefaniak and ewa król department of plant pathology, agricultural university, leszczyńskiego 7, pl-20-069 lublin zofia.machowicz@ar.lublin.pl; ewa.krol@ar.lublin.pl m a c h o w i c z s t e f a n i a k z., k r ó l e.: characterization of phoma negriana thüm., a new species from grapevine canes. acta mycol. 42 (1):113-117, 2007. the occurrence and elements of morphology of p. negriana thüm. were studied. the fungus cultures were isolated from grapevine canes cultivated in south–east poland. grapevine stems from which the cultures of p. negriana were obtained showed symptoms of necrosis and bark crashes. the species of the fungus was identified on the basis of pycnidia and conidia morphology, the character of colonies growth and biochemical features of the studied isolates. key words: phoma negriana, occurrence, elements of morphology introduction fungi from the genus phoma are represented by numerous species grouped into nine sections, selected on the basis of pycnidia structure, especially their cells wall, the presence of chlamydospores including dictiochlamydospores and the appearance of pycnidiospores (b o e r e m a 1997; d e g r u y t e r , n o o r d e l o o s , b o e r e m a 1998; d e g r u y t e r , b o e r e m a , va n d e r 2002). phoma spp. occur commonly in different geographical regions of the world on the plants from va rious botanical groups (s u t t o n 1980; m a r c i n k o w s k a 1995). considering manner of life and trophism within phoma spp. saprotrophs, opportunistic pathogenes and specific pathogenes of plants can be distinguished. among opportunistic pathogenes numerous species occur on above ground parts of fruit-trees and bushes. one of them is p. pomorum thüm., section peyronella, causing necrotic lesions on the leaves and fruit of apple tree (b o e r e m a 1993; b l e c h t o v a 1995). phoma macrostroma mont., sec. phyllostictoides, belongs to the complex of fungi colonizing decayed stems and buds of walnut, ash and beech (p i n t e r et al. 2000; g r i f f i t h , b o d d y 1990; to t i et al. 1993). phoma exigua var. populi de gruyter et scheer, sec. phyllostictoides was recognized as opportunistic pathogen of poplar seedling, p. exigua var. viburni (roum. ex sacc.) boerema as pathogen of various cultivars of viburnum acta mycologica vol. 42 (1): 113-117 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 114 z. machowicz-stefaniak and e. król and p. exigua var. lilacis (sacc.) boerema of lilac stems (d e g r u y t e r , s c h e e r 1998). phoma herbarum westend., sec. phoma, in addition to phomopsis viticola and cytospora sp. was isolated from grapevine stems with symptoms of bark necrosis (s t o j a n o v i č 1086). recently, phoma negriana thüm., sec. phoma has been recognized as opportunistic pathogen of grapevine in the netherlands (d e g r u y t e r et al.1998). this species was noted in the vineyards of south europe earlier, causing disease symptoms on the leaves, fruit and stems (d e g r u y t e r et al. l.c.). in poland p. negriana has not been observed so far. material and methods four isolates of p. negriana i.e. w 1205, w 1308, w 1075, w 1432 from a collection of this species isolates in our possession, were randomly chosen for the study. these cultures were isolated from grapevine canes cv. schuyler, iza and rf-16 with symptoms of necrosis and cultivated in south–east poland in the years 2000-2003. the occurrence of the fungi was determined on the basis of etiological symptoms present on the surface of diseased canes and on the basis of mycological analysis according to the artificial culture method. the isolation was carried out from superficially disinfected canes using maltose medium (biomerieux) according to m a c h o w i c z –s t e f a n i a k and k u r o p a t w a (1993). the obtained isolates of p. negriana were identificated on the standard media according to a monograph by d e g r u y t e r et al. (l.c.), with regard to the present principles of taxonomy (b o e r e m a 1976; m a r c i n k o w s k a 1995; b o e r e m a et al. 2004). the inoculum of chosen isolates of p. negriana was put in the centre of petri dishes with solidified media: malt-extract agar (ma), cherry agar (ca) and oatmeal mealagar (oa) (d e g r u y t e r , n o o r d e l o o s 1992). the cultures were incubated in thermostat for 7 days at temperature 22°c, without light and during a second week at 13 hours in uv light and 11 hours in the darkness. a description of the colonies morphology, with regards to the principles of b o e r e m a et al. (2004), and the measurment of their diameter were performed after 7 and 14 days of colonies incubation in the thermostat. after 2 weeks of incubation on oa the measurment of 400 conidia (4 isolates x 100 conidia) and 200 pycnidia (4 isolates x 50 pycnidia) were made and the presence of chlamydospores was tested. results the studies indicated that on the surface of grapevine canes in the place of necrotic lesions the pycnidia with conidia with features typical of phoma species were present. a big number of phoma spp. cultures were isolated from these canes. the share of phoma negriana isolates among fungi isolated from grapevine canes in the study years was: 0,4% in 2000; 1,4% in 2001; 0,3% in 2002 and 0,5% in 2003 (0,65% in average). the studies of selected isolates on the standard media indicated that 7-day-old fungus colonies on the ca medium were rather regular and had the diameter from 28 to 35 mm. in the beginning, they formed soft floccose, white–gray aerial mycelium with cream reverse. after 4-6 days of incubation in the centre of colony the pycnidia appeared and after 7 days conidia were formed. after 14 days of growth, the margin phoma negriana 115 of colonies was irregular and their diameter ranged from 44 to52 mm (fig. 1). the aerial mycelium was dark–green or gray. on the whole surface of the colony abundant pycnidia were formed. they were partly immersed in the substrate and secreted the yellow–saffron drops containing conidia. the reverse of 14-day-old colonies was dark–green to almost black. on oa medium, 7-old-days colonies had a less regular margin and colony diameter from 25 to 28 mm. the aerial mycelium was white–gray and floccose with a rather compact structure. first pycnidia were formed after 4 – 6 days of incubation. after 14 days the colony’s diameter was from 35 to 48 mm. the mycelium structure was slightly velvety, green–olivoceous with white margin. pycnidia were formed on the whole surface of the colony. on ma medium, the diameter of 7-day-old colonies was from 18 to 22 mm. the colonies were regular with floccose, white–gray surface. the diameter of 14-day-old colonies was from 23 to 35 mm. the colonies were with more or less regular growth, rather compact, of floccose structure, gray–green or gray–black. the reverse was olivaceous–black with a somewhat brighter margin. the formation of pycnidia was poorer than on oa and ca media. a detailed observation carried out on oa medium allowed to determine that pycnidia of the studied isolates of fungus were formed as singular or with small groups. they were thin–walled, globose or sometimes oval, honey–brown with papillate ostiole (tab. 1, figs 2, 3). the majority of pycnidia had 1 ostiole. the surface of the walls was smooth with some hyphal outgrowths. the diameter of the pycnidia was from 72 to 250 μm. conidia aseptate, oblong with 2 or more guttules (tab. 1, figs 4, 5). the conidia had length from 4.2 to 7.88 μm with some reaching 9.88 μm (tab. 1). the width of conidia ranged from 1.97 to 3.9 μm. chlamydospores and crystals were not observed in the cultures of the studied isolates of p. negriana. ta b l e 1 characterization of pycnidia and conidia of phoma negriana (mean for 4 isolates) author pycnidia conidia shape dimension in µm shape dimension in µm own data globose or oval with papillate ostiole 72 250 oblong, usually with 2 or more guttules 4.2 7.88 (9.85) x 1.97 3.9 de gruyter et al. 1998 globose or irregular, solitary or confluent, glabrous with 1-2 (4) papillate ostiole(s) 70 220 ellipsoidal to oblong, with several distinct guttles. 4.5 8.5 (10.5) x 2 4 in the case of 3 isolates no changes were observed in colony coloration on the media ma and oa after reaction with 1n naoh. only in the case of isolate w 1432 on ma medium the change of coloration into red–brown was noted. 116 z. machowicz-stefaniak and e. król discussion the obtained results concerning characterization of colonies growth of the studied isolates, the morphology of pycnidia and conidia, compared with a description in a monograph of phoma section (d e g r u y t e r et al. l.c.; b o e r e m a et al. 2004) allowed to identify the studied species as phoma negriana. macroscopic features of colonies of the studied isolates of p. negriana, especially the growth rate, colouration and mycelium structure showed similarity to a description by d e g r u y t e r et al. ( l.c.). the observation indicated that the mentioned feature and character of the colonies growth should have great importance in identification of the species from the genus phoma which corresponds to earlier information of other authors (b o e r e m a 1976; d e g r u y t e r , n o o r d e l o o s 1992; d e g r u y t e r et al. l.c.). considering great morphological similarity of conidia within phoma species, their dimension should be an important but additional diagnostic feature (b o e r e m a 1997; m a r c i n k o w s k a 1995; z i m o w s k a , m a c h o w i c z -s t e f a n i a k 2005). according to the opinion by d e g r u y t e r et al. (l.c.) the reaction of p. negriana isolates with 1n naoh is not specific, which confirms the results of the present study. the fact of isolation of p. negriana isolates from grapevine canes contributs to an increased a number of species described on the stems of this plant, cultivated in south–east poland. considering earlier recognition of this fungus as opportunistic pathogen of the above ground parts of grapevine (d e g r u y t e r et al. 1998), the possible pathogenicity of the fungus towards this plant in climatic conditions of our country require more scrupulous tests. references b l e c h t o v a a. 1995. ability of some micromycetes of cause necroses of apple tree bark in vitro. ochr. rostl. 31: 265–276. b o e r e m a g. h. 1976. the phoma species studied in culture by dr. r.w.g. dennis. trans. brit. mycol. soc. 67: 289–319. b o e r e m a g. h. 1993. contributions towards a monograph of phoma (celomycetes) ii section peyronellaea. persoonia 15: 187–221. b o e r e m a g. h. 1997. contributions towards a monograph of phoma (celomycetes) v. subdivision of the genus in section. mycotaxon 64: 321–333. b o e r e m a g. h., g r u y t e r j. de, n o o r d e l o o s m. e., h a m e r s m. e. c. 2004. phoma identification manual. differentiation of specific and intra-specific taxa in culture. cabi publishing, cambridge, usa, 470 pp. g r i f f i t h g. s., b a d d y l. 1990. fungal decomposition of attached angiosperm twigs. i. decay community development in ash, beech and oak. new phytologist 116: 407–415. g r u y t e r j. de, n o o r d e l o o s m. e. 1992. contributions towards a monograph of phoma: taxa with very small conidia in vitro. persoonia 15: 71–92. g r u y t e r j. de, n o o r d e l o o s m. e., b o e r e m a g. h. 1998. contributions towards a monograph of phoma (celomycetes) i. 3. section phoma: taxa with conidia longer than 7 µm. persoonia 16: 471– 490. g r u y t e r j. de, s c h e e r p. 1998. taxonomy and pathogenicity of phoma exigua var. populi var. nor. causing necrotic bark lesions on poplars. j. phytopathology 146: 411– 415. g r u y t e r j. de, b o e r e m a g. h., va n d e r aa h. a. 2002. contributions towards a monograph of phoma (celomycetes) vi-2. section phyllostictoides: outline of its taxa. persoonia 18: 1–53. m a c h o w i c z s t e f a n i a k z., k u r o p a t w a e. 1993. grzyby porażąjące winorośl uprawianą pod osłonami. mat. xxxiii sesji nauk. ior. ii-postery, poznań: 169 –173. phoma negriana 117 m a r c i n k o w s k a j. 1995. nowoczesne spojrzenie na systematykę rodzaju phoma. wiad. bot. 39: 47– 52. p i n t e r c., f i s c h l g., k a d l i c s k o s., d a n k o j., g a r a m., m a k o s. 2000. pathogenes of walnut in hungary. proceedings 52 nd international symposium on crop protection, gent, belgium, 9 may, part ii. mededelingen faculteit landbouwkundigun toegepaste biologische wetenschappen, universiteit 65: 673–676. s t o j a n o v i č s. 1986. microflora suwih lostra vinove loze. zaščita bilja 37: 153–186. s u t t o n b. c. 1980. the coelomycetes. fungi imperfecti with pycnidia, acervuli and stroma. commonwealth mycological institute, kew, surrey, england, 696 pp. to t i l., v i r e t o., h o r a t g., p e t r i n i o. 1993. detection of the endophyte discula umbrinella in buds and twigs of fagus sylvatica. eur. j. for path. 23: 147–152. z i m o w s k a b., m a c h o w i c z s t e f a n i a k z. 2005. charakterystyka izolatów phoma strasseri nie notowanego w polsce patogenu mięty pieprzowej (mentha piperita l.). acta agrobot. 58 (2): 151–163. charakterystyka phoma negriana thüm, nowego gatunku z łozy winorośli s t r e s z c z e n i e przebadano cztery losowo wybrane izolaty phoma negriana z własnej kolekcji kultur tego gatunku. izolaty wyosobniono w latach 2000-2003 z łozy winorośli odmian schuyler, iza i rf-16 z objawami nekrozy i pękania kory. oznaczano je na pożywkach standardowych: maltozowa (ma), wiśniowa (ca) i owsiana (oa), przy uwzględnieniu aktualnych zasad taksonomii phoma spp. opis morfologii kolonii oraz pomiar ich średnicy wykonano po 7 i 14 dniach hodowli. po 2 tygodniach wzrostu na pożywce oa wykonano pomiar 400 konidiów, 200 piknidiów oraz sprawdzono obecność chlamydospor. uzyskane wyniki upoważniły do uznania badanego gatunku jako phoma negriana thüm., sekcja phoma. makroskopowe cechy kolonii badanych izolatów, zwłaszcza tempo i charakter wzrostu, zabarwienie i struktura grzybni powinny mieć istotne znaczenie przy identyfikacji gatunków z rodzaju phoma. ze względu na duże podobieństwo morfologiczne zarodników phoma spp., ich wymiary należałoby traktować jako ważną, aczkolwiek pomocniczą cechę diagnostyczną. uzyskane wyniki potwierdziły niespecyficzny charakter reakcji izolatów p. negriana z 1 n naoh. w polsce badany gatunek nie był dotychczas notowany. fig. 1. 14-day-old colonies of phoma negriana, w 1075 on the standard media. phot. e. król. fig. 2. pycnidia of phoma negriana, w 1075, 160 x magnification. phot. e. król. fig. 3. pycnidia of phoma negriana (sem), 200 x magnification. phot. m. wróbel. fig. 4. conidia of phoma negriana, 640 x magnification. phot. e. król. fig. 5. conidia of phoma negriana (sem), 4300 x magnification. phot. m. wróbel. 2014-01-01t11:45:35+0100 polish botanical society attempts at active protection of inonotus obliquus by inoculating birches with its mycelium jacek piętka and andrzej grzywacz department of mycology and forest phytopathology,warsaw agricultural university nowoursynowska 159, pl 02 776 warszawa jacek_pietka@sggw.pl, andrzej_grzywacz@sggw.pl p i ę t k a j., g r z y w a c z a.: attempts at active protection of inonotus obliquus by inoculating birches with its mycelium. acta mycol. 41 (2): 305 312, 2006. practical application of active protection methods of inonotus obliquus (fr.) pilát. was examined. thirty live birches and 15 birch stem sections were artificially inoculated with the fungal mycelium in the mińsk forest district (e poland). the mycelium of i. obliquus was not recorded in the felled test trees and birch stem sections upon the completion of the experiment. artificial introduction of i. obliquus in the natural environment faces significant problems caused by strong competition from other birch wood decay fungi. as in vitro studies show (individual biotic effect determination), the fungi examined, occurring on birch trees in nature, are dominant species in relation to i. obliquus. key words: active protection, inonotus obliquus, tree inoculation, liginicolous fungi, birch, competition introduction inonotus obliquus (fr.) pilát is a fairly rare parasitic fungus developing primarily on birch trees: betula pendula, b. pubescens, b. carpatica, as well as infrequently on maple trees: acer campestre, a. pseudoplatanus, alder trees: alnus glutinosa, a. incana, oak trees: q. cerris, q. petraea, ash trees fraxinus excelsior, elm trees ulmus, beech trees fagus, poplar trees populus, rowan trees sorbus, hophornbeam trees ostrya (k o t l a b a 1984; s e m e n k o v a , s o k o l o v a 1992; r y v a r d e n , g i l b e r t s o n 1993; z a b e l 1947). in poland, where it decays almost exclusively birch wood, the fungus seems to be more common in podlasie, including the białowieża forest, as well as in masuria and great poland (m a ń k a , s t u b e 1952; d o m a ń s k i 1965). it is considered to be rare in upper silesia (wo j e w o d a 1999) and the góry świętokrzyskie mts. (ł u s z c z y ń s k i 2002). outside europe, it is also encountered in asia and north america (canada, usa) (k o t l a b a 1984). acta mycologica vol. 41 (2): 305-312 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 306 j. piętka and a. grzywacz the fungus causes extensive white rot of wood, most often infecting trees aged 30-50. infection of healthy trees occurs through wounds, knots, frost shakes. after a few years, the parasite produces vegetative fruitbodies (sterile conks): irregular black lumps (knobs, growths) on live trees, usually in the place of the initial infection. the parasite may develop on a trunk for 30-80 years (č e r n ý 1976). fruitbodies may weigh up to 3-5 kg after 10-15 years (g a m m e r m a n et al. 1975), and specimens as heavy as 16 kg have been reported (s i n a d s k i j 1973). the diameter of old fruitbodies on thick old trees may even reach 0.5 m (d o m a ń s k i 1965), their thickness 10-15 cm, and length – 1-1.5 m (g a m m e r m a n et al. 1975). the surface of the sterile conk is strongly cracked, very hard, brittle, black, as if charred; its flesh is also hard, brown, with yellowish discolouration. numerous chlamydospores that enable infection of other trees develop inside the sterile conk until the host’s death (d o m a ń s k i 1965). the fungus was for a long time considered to be a sterile form of fomes igniarius f. sterilis van. (phellinus igniarius f. nigricans (fr.) bond.) (va n i n 1955). in poland, sterile conks most often occur on birch trunks up to 9 m, in all the directions of the world, preferring, however, the north, north-east and north-west: fruitbodies develop best on the less insolated side where humidity conditions are more favourable. more vegetative fruitbodies were recorded in older age class treestands, and the dimensions of individual collected specimens were greater on trees with a bigger mean diameter at breast height (dbh) (p r z e s ł a w 1985). resupinate generative fruitbodies appear when the tree is dying or after it has died, under the bark of standing or lying trees. such fruitbodies occur only once in august – september, always on the trunk side where the rot is most advanced, often in the place of sterile conks or where their remnants are visible (č e r n ý 1976). generative fruitbodies sometimes reach very big dimensions: 3-4 m long and up to 50 cm wide (b o n d a r c e v a , p a r m a s t o 1986). their margin is usually thicker (2.5-4 mm), and forces the bark apart from the wood, allowing the fruitbody to expand. tubes are always positioned obliquely to the substrate, usually at 20-30º (hence the species name). young tubes are yellow-olive, older – rusty-brown. pores are polygonalspherical, often elongated, 3-4 in 1 mm (d o m a ń s k i 1965). insects often quickly eat generative fruitbodies, making it difficult to find them (r y v a r d e n , g i l b e r t s o n 1993). in the us, i. obliquus produced a generative fruitbody on an infected beech tree (fagus grandifolia) that was still alive (z a b e l 1947). an appropriately prepared extract from sterile conks has medicinal properties. it was used in cancer, intestinal pains, hyperacidity, gastric and duodenal ulcerations, spleen and liver disorders, and applied externally in the inflammation of the oral cavity and reproductive organs (p i a s k o w s k i 1957; o ż a r o w s k i 1980; g r o c h o w s k i 1992). in the former states of the soviet union, it is still used extensively in several disorders. a number of medicinal products, including befungin and binczaga, thickened fungal extracts with added cobalt salt, were prepared at the leningrad botanical institute. the fungus has a favourable effect on the central nervous system and metabolic processes, and boosts immunity to infections. aqueous extracts have been shown to greatly alleviate the suffering in cancer patients, relieve pain, improve appetite; it is not, however, a radical drug in malignant cancer cases, although it inhibits the development of the disease if used at its initial stages (s a u t i n et al. 1984). active protection of inonotus obliquus 307 sterile conk infusions are drunk instead of tea in siberia (c a r t w r i g h t , f i n d l a y 1951; s a u t i n et al. 1984). i. obliquus parasitizes various deciduous trees; only fruitbodies on birch trunks, however, have medicinal properties. they may be collected throughout the year; winter and early spring are most convenient as it is easier to find sterile conks on leafless trees (s a u t i n et al. 1984). i. obliquus is one of the 20 fungal species to be monitored in poland within the national environmental monitoring programme adopted in may 1992. selected also because of its medicinal properties, the fungus may be noticed and recognised easily in the field by non-specialists (g r z y w a c z et al. 1997; ł a w r y n o w i c z 2000). it is red-listed as r (rare) on the “red list of macrofungi in poland” (wo j e w o d a , ł a w r y n o w i c z 2006). it is partially protected pursuant to the regulation of the minister of the environment of 2004 on protected species of wild-growing fungi; fruitbodies may be acquired only with the voivode’s permission (dz. u. 2004.168.1765). the aim of this study was to examine practical application of active protection methods and to artificially cultivate i. obliquus, a species with medicinal properties, which has been excessively exploited and is now disappearing and becoming increasingly rare. the feasibility of artificial infection of live trees and dead birch wood with the mycelium of i. obliquus, acquired in natural conditions and cultivated in laboratory conditions, was examined. biotic relationships between i. obliquus and selected fungi occurring on birch trees were investigated to assess the competition degree and their inhibitory function. materials and methods a sterile conk of i. obliquus was obtained from a live birch tree, betula pendula (dbh 26 cm), growing in a tree-stand in the mińsk forest district (e poland). the fruitbody, 8 cm wide, 8 cm long and 12 cm tall, was cut out with a large wood fragment and transported to the laboratory of the department of mycology and forest phytopathology, warsaw agricultural university, where a pure mycelium culture was isolated from fragments of infected wood directly beneath the fruitbody. the individual biotic effect (ibe) determined using plate tests. petri dishes 7 cm in diameter with agart-wort medium were used. two fungal species were inoculated 2 cm apart in the central part of each plate: i. obliquus and a fungus belonging to a specific community (competitive species). competitive species were substituted in successive tests, repeated 10 times; the fungus examined was used in each test. competitive species and i. obliquus were also inoculated separately, each variant repeated 5 times. the mutual influence of i. obliquus and the fungi occurring in the same trophic environment, weakened birch trees and dead wood (stem sections, stumps), was examined in the experiment, i.e.: daedaleopsis confragosa, fomes fomentarius, fomitopsis pinicola, lenzites betulina, piptoporus betulinus, trametes versicolor. pure fungal cultures from the collections of the department of mycology and forest phytopathology were used. plates were incubated in a heraeus incubator at 22°c. monocultures and dicultures were measured after 10 days. the development of dicultures served to determine the “individual biotic effect”, using the evaluation scale established by m a ń k a (1974). as given in the method, a positive ibe shows an inhibiting influence. 308 j. piętka and a. grzywacz artificial infection of stem sections and trunks in natural sites. a field experiment was established in september 1999: birch stem sections and selected, healthy birch trees were inoculated. in the first part of the experiment, the wood of a felled birch was cut into stem sections, 60 cm long, and left to dry. after 11 days, they were inoculated at the height of 30 cm, on 4 sides, with birch wood inocula, 30 x 5 x 5 mm (50%), and parts of sambucus nigra shoots (50%) overgrown with the mycelium of i. obliquus. the procedure was used as the mycelium of i. obliquus colonised dead birch wood inocula less intensively while it easily overgrew the stems of s. nigra shoots. the inoculated stem sections (15 pieces) were placed in two rows under the crowns of fruit trees, slightly sinking their lower parts in the soil. the experiment was established in a plot in the village of maliszew near mińsk mazowiecki. in the second part of the experiment, 30 live birches were inoculated in the mińsk forest district (section 317 a), in the vicinity of the cegłów-podskwarne road. as specified in the forest management plan of the mińsk forest district (1996-2005), the birches were 69 years old. they were inoculated at two levels, 2 and 3 m, in the four directions of the world. inocula consisting of s. nigra shoots overgrown with the mycelium of i. obliquus were used at 2 m while birch wood inocula overgrown with the mycelium were used at 3 m. the mean dbh of the inoculated birches was 35.5 cm. the trees were marked permanently with galvanised iron plates. the permission of the head forester in the mińsk forest district was obtained to establish the experiment. holes 10-12 cm deep and 8 mm in diameter were drilled with a battery drill. one inoculum was introduced into each hole with tweezers and closed up with freshly obtained shoots of corylus avellana. the experiment was terminated in 2002. the stem sections were split along the line delineated by the places of visible inlet holes. wood fragments, appropriately labelled and packed in paper envelopes, were transported to the laboratory. two live inoculated trees were also obtained. two one-meter long trunk sections were made from each birch tree (from the height at 1.5-2.5 m and 2.5-3.5 m) and transported to the laboratory where they were split. the initial inoculum was removed in the inoculation chamber. wood samples were collected at 1 cm from the inoculation point and placed on agart-wort medium in petri dishes. if visible rot was observed, the wood was sampled at successive distances along the trunk axis every 2 cm. results the findings show that i. obliquus is inhibited by the birch wood-decay fungi examined (tab.1). the individual biotic effect is influenced mostly by competition. the inhibition zone was not recorded in any experiment variant. the highest ibe index +5 was recorded for l. betulina and the lowest (ibe +2) for d. confragosa. the results of artificial inoculation of trees and stem sections with the mycelium of i. obliquus were examined in october 2002, 3 years after the experiment had been established. fruitbodies of the following fungal species were observed on the 15 birch stem sections: trametes versicolor on 9 stem sections, stereum hirsutum on 3 stem sections, active protection of inonotus obliquus 309 bjerkandera adusta on 3 stem sections. i. obliquus was not isolated in the laboratory from the birch stem sections artificially infected with its mycelium. two trees were additionally felled in the study area in the mińsk forest district (tree no 19, dbh 29 cm; tree no 24, dbh 28 cm). the felled birches had distinct star false heartwood, whose arms overlapped with the inlet hole points through which the inocula had been inserted (figs 1, 2). re-isolation of the mycelium of i. obliquus from these trees failed. a big participation of moulds and the mycelium of undetermined basidiomycetes (hyphae with clamp-connections) was, however, observed during the isolation on media in petri dishes. the visible star heartwood in birch no 19 stretched from the height of 0.5 m up to 5.7 m while it was still slightly visible at the level of the felling (ca. 10 cm.) and reached up to 6 m in birch no 24. discussion the mycelium of i. obliquus was not reisolated from either the stem sections or live trees after the 3 years of the experiment. fruitbodies of some other fungal species were recorded on the stem sections: trametes versicolor, stereum hirsutum, bjerkandera adusta. this seems to confirm the claim that i. obliquus develops exclusively on live trees. dead birch wood was thus naturally colonised by common saprotrophic species. distinctive, dark-coloured star false heartwood whose arms overlapped with inlet hole points was recorded in the felled test trees. k r z y s i k (1974) claims that false heartwood occurs as a result of the penetration and destructive activity of fungi or external factors such as heavy frost or air penetration, and notices that heartwood is irregular or star-like in the areas adjacent to air penetration points. heartwood compounds that are products of the dying parenchymal cell content penetrate vessels and adjacent tissues, producing brown discolouration (k r z y s i k 1974). as observed during laboratory trials to re-isolate the mycelium of i. obliquus, other undetermined basidiomycetes (clamp-connections visible on the mycelium) that may have contributed to the development of false heartwood were recorded. other fungal spores may also have penetrated the trunk when the experiment was established or later through the inlet holes. having germinated, they found good development conditions and inhibited the growth of the mycelium of i. obliquus. p i ę t k a and g r z y w a c z (2005) report that positive effects were obtained in an experiment in which live old larches ta b l e 1 influence of selected fungal cultures colonising birch wood on the development of the mycelium of i. obliquus fungal species rot type individual biotic effect (ibe) index daedaleopsis confragosa white +2 fomes fomentarius white +4 fomitopsis pinicola brown +4 lenzites betulina white +5 piptoporus betulinus brown +4 trametes versicolor white +4 310 j. piętka and a. grzywacz were inoculated with the mycelium of fomitopsis officinalis. an active mycelium was observed in both felled test trees after 3 years of the experiment. a strong resin reaction of the inoculated larches was observed on the day when the experiment was established. an outward resin leak from the sapwood was advantageous as the inlet hole was sealed up and potential infection by other organism was limited. visible false heartwood in the felled birches reached from ca. 0.1-0.5 m (1.5-1.9 m downwards from the level of the lowest holes) to 5.7-6m (2.7-3 m upwards from the highest holes), which shows that it occurs significantly more quickly up the trunk. the ibe results (tab. 1) of various birch wood-decay fungi show that i. obliquus is an inhibited species. laboratory examinations (on agar-wort medium in petri dishes) show that fungal competition yields relatively clear results; the results of these examinations, however, may not always be applied to relationships between these fungi in natural conditions, on a specific host plant. s c h w a r z e et al. (2000) report that the infection success depends on a number of factors: parasitic abilities of the fungus, tree vitality, type and size of the injury, environmental factors (such as temperature, humidity, oxygen content in the substrate), pathogen’s morphological specialisation. it should also be remembered that some fungal species exhibit strain diversification, exhibited in the value of the ibe, as observed by m a ń k a (1999) or ty s z k i e w i c z and m a ń k a (1999). the infection success is affected not only by competitive fungi of the class basidiomycetes but also by a number of other fungi (moulds, fungi causing wood stains) and bacteria. only detailed examinations of the quantitative and qualitative structure of the fungal communities colonising knots and mechanical injuries of the birches would help determine the actual function of these communities in relation to i. obliquus and to assess potential successful infection. it may be supposed, however, that the inhibition of the i. obliquus mycelium growth in laboratory examinations by other basidiomycetes encountered on weakened birches may to some extent contribute to the number of successful infections by this fungal species in nature. it may not be definitely concluded that the mycelium of i. obliquus did not colonise the other birches, artificially infected in this experiment. sterile conks may in the future become visible on the other test trees in the birch wood in the mińsk forest district, which would help assess the success rate of active protection methods of this fungus be means of artificial infection of live trees. acknowledgements. we are grateful to dr. andrzej szczepkowski for his assistance in establishing the experiments and collecting research material. conclusions 1. as a parasitic species that colonises only live trees in nature, inonotus obliquus did not colonise artificially infected birch stem sections. it was primarily colonised by the following species: trametes versicolor, stereum hirsutum, bjerkandera adusta. 2. attempts to artificially introduce inonotus obliquus to the natural environment face great problems caused by strong competition from other birch wood-decay fungi. the results of individual biotic effects show that inonotus obliquus may lose the competition for the nutritive basis (weakened birches) with fungi occurring in nature. active protection of inonotus obliquus 311 3. the recorded false heartwood in the test trees developed almost twice as fast up the trunk from the inlet hole points. 4. the examinations conducted show that given the current knowledge on the subject there are no practical possibilities of active protection of inonotus obliquus. other methods of artificial inoculation of birches should be devised. the mycelium may be stored as pure cultures in a laboratory. references b o n d a r c e v a m. a., p a r m a s t o é. ch. 1986. opredelitel΄ gribov sssr. porjadok afilloforovye. le ningrad, nauka. c a r t w r i g h t k. s. g., f i n d l a y w. p. k. 1951. rozkład i konserwacja drewna. pwril, warszawa. č e r n ý a. 1976. lesnická fytopatologie. státni zemĕdĕlské nakladatelstvi, praha. d o m a ń s k i s. 1965. flora polska. grzyby (fungi) 2: basidiomycetes. aphyllophorales. polyporaceae i. mucronoporaceae i, pwn, warszawa. dz.u.2004.168.1765. rozporządzenie ministra środowiska z dnia 9 lipca 2004 r. w sprawie gatunków dziko występujących grzybów objętych ochroną. g a m m e r m a n a. f., k a d a e v g. n., š u p i n s k a j a m. d., j a c e n k o c h m e l e v s k i j a. a. 1975. lekarstvennye rastenija. vysšaja škola, moskva. g r o c h o w s k i w. 1992. włóknouszek ukośny w przyrodzie i medycynie. głos lasu, 12: 28 29. g r z y w a c z a., b u j a k i e w i c z a., ł a w r y n o w i c z m., w o j e w o d a w. 1997. monitoring przyrody żywej. grzyby wielkoowocnikowe. warszawa, (msc.). k o t l a b a f. 1984. zemĕpisne rozšiřeni a ekologie chorošu (polyporales s. l.) v československu. acade mia praha. k r z y s i k f. 1974. nauka o drewnie. pwn, warszawa. ł a w r y n o w i c z m. 2000. podstawy monitoringu grzybów w polsce (in:) m. l i s i e w s k a , m. ł a w r y n o w i c z (eds). monitoring grzybów. polskie towarzystwo botaniczne, poznań łódź: 9 15. ł u s z c z y ń s k i j. 2002. preliminary red list of basidiomycetes in the góry świętokrzyskie mts (poland). polish botanical journal 47 (2): 183 193. m a ń k a k., s t u b e t. 1952. występujący na brzozach grzyb poria obliqua (pers.) bres. i jego rozwój na sztucznych pożywkach. acta soc. bot. pol., 21: 517 536. m a ń k a k. 1974. zbiorowiska grzybów jako kryterium oceny wpływu środowiska na choroby roślin. phy topathologia polonica. zeszyty problemowe postępów nauk rolniczych, pwn. 160: 9 135. m a ń k a m. 1999. taksonomia i fitopatologiczna funkcja saprofitycznych grzybów glebowych środowiska leśnego. zeszyty naukowe ar w krakowie, 63. o ż a r o w s k i a. (ed.) 1980. ziołolecznictwo. państwowy zakład wydawnictw lekarskich, warszawa. p i a s k o w s k i s. 1957. wstępne badania nad otrzymywaniem i zastosowaniem preparatów z czarnej huby brzozowej w przypadkach nowotworów złośliwych u ludzi. sylwan ci, 10: 5 12. p i ę t k a j., g r z y w a c z a. 2005. in situ inoculation of larch with the threatened wood decay fungus fomitopsis officinalis (basidiomycota) experimental studies. polish botanical journal 50 (2): 225 231. p r z e s ł a w b. 1985. biologia włóknouszka ukośnego (inonotus obliquus) i jego występowanie w drzewo stanach brzozowych. master’s thesis (msc.). katedra ochrony lasu i ekologii sggw, warszawa. r y v a r d e n l., g i l b e r t s o n r. l. 1993. european polypores. part 1,2. fungiflora, oslo. s a u t i n v. i., f o m i n a v.i., va l o v a z.g. 1984. dary našich lesov. izdatel΄stvo polymja, minsk. s c h w a r z e f. w. m. r., e n g e l s j., m a t t h e c k c. 2000. fungal strategies of wood decay in trees. springer verlag, berlin, heidelberg, new york. s e m e n k o v a i. g., s o k o l o v a é. s. 1992. lesnaja fitopatologija. ékologija, moskva. s i n a d s k i j j. v. 1973. bereza eë vrediteli i bolezni. izdatel΄stvo nauka, moskva. ty s z k i e w i c z z., m a ń k a m. 1999. wpływ zbiorowisk grzybów glebowych dwu grądów na wzrost in vitro potencjalnych patogenów korzeni drzew. zeszyty naukowe ar w krakowie 63. va n i n s. i. 1955. lesnaja fitopatologija. goslesbymizdat, moskva leningrad. w o j e w o d a w. 1999. czerwona lista grzybów wielkoowocnikowych górnego śląska. raporty opinie 4, centrum dziedzictwa przyrody górnego śląska. 3. katowice 312 j. piętka and a. grzywacz w o j e w o d a w., ł a w r y n o w i c z m. 2006. red list of macrofungi in poland. (in:) z. m i r e k , k. z a r z y c k i , w. w o j e w o d a , z. s z e l ą g (eds). red list of plants and fungi in poland. w. szafer inst. bot., pol. acad. sci., kraków: 55 70. z a b e l r. a. 1947. poria obliqua on dying beech. phytopathology 37: 189 190. próby ochrony czynnej inonotus obliquus przez szczepienie brzóz jego grzybnią s t r e s z c z e n i e błyskoporek podkorowy (włóknouszek ukośny) inonotus obliquus (fr.) pilát. to niezbyt częsty grzyb pasożytniczy rozwijający się w polsce przede wszystkim na brzozach, powodując rozległą białą zgniliznę drewna. po kilku latach sprawca wytwarza na żywych drzewach, z re guły w miejscach pierwotnej infekcji owocniki wegetatywne, przybierające kształty nieregular nych czarnych brył (guzów, czyrów). odpowiednio przyrządzony wyciąg z owocników wegetatywnych wykazuje właściwości lecznicze. w wielu krajach do tej pory owocniki te stosowane są bardzo szeroko, na wiele różnych schorzeń. i. obliquus w 2004 roku objęty został w polsce ochroną częściową, znajduje się również na „czerwonej liście grzybów wielkoowocnikowych zagrożonych w polsce”, gdzie zapisany jest w kategorii r gatunków rzadkich. w pracy tej starano się ustalić, czy istnieje możliwość sztucznej infekcji żywych drzew i martwego drewna brzozowego grzybnią i. obliquus pozyskaną w warunkach naturalnych i wyhodowaną w warunkach laboratoryjnych. badano również stosunki biotyczne pomiędzy i. obliquus a wybranymi grzybami występującymi na brzozie, w celu ustalenia w jakim stopniu stanowią konkurencję i czynnik ograniczający dla wzrostu grzybni i. obliquus. w 1999 roku zaszczepiono 30 żywych brzóz oraz 15 wałków brzozowych inokulatami z grzybnią na terenie nadleśnictwa mińsk. po 3 latach zlikwidowano doświadczenie dotyczące wałków. okazało się, iż martwe drewno brzozowe było naturalnie kolonizowane przez pospo lite gatunki saprotroficzne: trametes versicolor, stereum hirsutum, bjerkandera adusta. pozy skano również 2 drzewa żyjące (z 30 zaszczepionych). w kabinie szczepień z przywiezionych fragmentów drewna wyjmowano pierwotne inokulum oraz pobierano wycinki z odległości 1 cm od miejsca inokulacji, które następnie kładziono na pożywkę agarowo brzeczkową w płytkach petriego. w przypadku zauważalnej zgnilizny wycinki drewna pobierano z kolejnych odległości wzdłuż osi pnia, co 2 cm. w ściętych drzewach próbnych oraz wałkach brzozowych nie stwierdzono obecności grzybni i. obliquus. próby sztucznego wprowadzania i. obliquus do środowiska naturalnego napotykają na duże problemy związane z silną konkurencją innych grzybów rozkładających drewno brzozowe oraz prawdopodobnie z innych powodów, nie do końca rozeznanych w tych doświadczeniach. stwierdzono, iż w pniach ściętych drzew prób nych wytworzyła się wyraźna, ciemno zabarwiona fałszywa twardziel o zarysie gwiaździstym, której ramiona pokrywały się z miejscami wykonania nawiertów. wyniki badań laboratoryjnych dotyczące indywidualnych efektów biotycznych pomiędzy różnymi grzybami rozkładającymi drewno brzozowe pokazują, iż i. obliquus jest gatunkiem ograniczanym. przeprowadzone badania wskazały, iż zastosowane metody nie dają praktycznych możli wości czynnej ochrony inonotus obliquus. należy poszukiwać innych metod sztucznego szcze pienia brzóz. 2014-01-01t11:44:45+0100 polish botanical society the participation of macromycetes in selected forest communities of the masurian landscape park (ne poland) grzegorz fiedorowicz department of mycology, university of warmia and mazury in olsztyn oczapowskiego 1a, pl-10-719 olsztyn-kortowo, grzegorz.fiedorowicz@uwm.edu.pl fiedorowicz g.: the participation of macromycetes in selected forest communities of the masurian landscape park (ne poland). acta mycol. 44 (1): 77–95, 2009. results of mycosociological studies in selected forests communities of the masurian landscape park between 1997 and 2000 are discussed. observations were conducted in 8 permanent plots and 69 supplementary plots (400 m2). five plant associations characteristic of the masurian landscape park, peucedano-pinetum, serratulo-pinetum, vaccinio uliginosipinetum, tilio-carpinetum and fraxino-alnetum, were examined. a total of 335 macromycete species were recorded. the greatest number of species was observed in tilio-carpinetum (198). key words: macromycetes, ascomycetes, basidiomycetes, forest communities, masurian landscape park introduction the masurian landscape park (mlp) was established in december 1977 in order to preserve and protect outstanding values of the natural environment of the masurian lake district. it aims to protect the richness of the fauna and the flora as well as the cultural and historical heritage. the park is of great research and teaching importance and is an interesting tourist and recreation area (dąbrowski, polakowski and wołos 1999). a comparatively clean environment, climatic conditions and the occurrence of a great variety of plant communities relatively unaffected by anthropogenic influence have contributed to the high diversity of fungi and the preservation of many fungal species threatened in poland or in europe. first reports on macromycetes from the area of the former german administrative districts of sensburg (mrągowo) and johonnisburg (pisz) date back to the early 20th century (abromeit 1905; neuhoff 1933). alina skirgiełło conducted wide acta mycologica vol. 44 (1): 77–95 2009 78 g. fiedorowicz ranging mycological observations in the vicinity of kamień and ruciane, that is in the area of the mlp at present, in the 1950s. participants of the iv congress of european mycologists collected fungi in the vicinity of mikołajki, kamień and ruciane in september 1966 (skirgiełło 1968; kotlaba, lazebniček 1967). studies on selected groups of macromycetes in the mlp have also been conducted by orłoś and dominik (1960), domański (1963), durska (1971), olesiński and wojewoda (1985). a total of 289 taxa of macromycetes have been reported from the mlp in the available literature. no data, however, are given on the phytosociological status of their localities. study area the masurian landscape park stretches between longitudes 21°20’ and 21°53’ e and latitudes 53°36’ and 53°51’ n. the surface area of the mlp is 53.655 ha and that of its protection zone is 18.608 ha. forests cover 50% of the area (27.140 ha), waters comprise almost 30% (15.995 ha) and arable lands constitute nearly 16.6% (8.920 ha) (polakowski, jutrzenka-trzebiatowski and hołdyński 1997). the mlp is situated on the border of three mesoregions in the physiogeographical regionalisation of poland. the western part of the park constitutes a fragment of the pojezierze mrągowskie lake district, the eastern part belongs to the kraina wielkich jezior mazurskich lake district, and its southern edges are part of the równina mazurska plain (kondracki 1998). the occurrence of numerous lakes is a feature that distinguishes the mlp from other protected areas. those are mostly large lakes whose surface area exceeds 50 ha. they comprise 22 lakes such as śniardwy, bełdany, mokre or łuknajno. a total of 60 lakes whose surface area is greater than 1 ha have been recorded in the mlp (bajkiewicz-grabowska 1989). the phytosociological composition of the mlp is greatly diversified. polakowski et al. (1976) distinguished 67 phytocoenoses in the rank of association or community based on their studies. peucedano-pinetum and serratulo-pinetum dominate among forest communities in the mlp (polakowski et al. 1997). material and methods mycosociological field studies were carried out between 1997 and 2000. three hundred and twelve observations were conducted in 8 permanent plots (from may 1997 to december 1999) and 106 observations in 69 supplementary plots in the plant associations examined (fig. 1). a total of 418 observations were carried out. observations at permanent plots were conducted every two or three weeks throughout the calendar year. observations were also made in supplementary plots established in additional patches of phytocoenoses. supplementary plots were observed between one and eight macromycetes in selected forest communities 79 times. the surface area of all observation plots was 400 m2 (lisiewska 1965; friedrich 2000). the commonly used braun-blanquet scale adjusted for fungi by moser was used in the quantitative assessment (nespiak 1959; lisiewska 1965). the following bioecological groups of fungi were distinguished based on the type and manner of substrate utilization: mycorrhizal fungi, fungi growing on humus, litter-inhabiting fungi, fungi on wood, bryophilous fungi and parasitic fungi (lisiewska 2000). the nomenclature of ascomycetes follows chmiel (2006) and hansen & knudsen (2000). the nomenclature of basidiomycetes follows wojewoda (2003). names of vascular plants are given according to mirek et al. (2002) and bryophytes according to ochyra et al. (2003). names of phytosociological units are admitted after matuszkiewicz (2001). the herbarium material is deposited in the herbarium of the chair of mycology, university of warmia and mazury, olsztyn. results and disscusion mycological observations were conducted in 8 permanent plots. they represented five plant communities: peucedano-pinetum (w. mat. 1962) w. mat. & j. mat. 1973, vaccinio uliginosi-pinetum kleist 1929, serratulo-pinetum (w. mat. 1981) j. mat. 1988, tilio-carpinetum tracz. 1962, fraxino-alnetum w. mat. 1952 (matuszkiewicz 2001) (tab. 1). fig. 1. location of permanent plots in the masurian landscape park. 1 – waters, 2 – forests, 3 – permanent research plots, 4 – complementary research plots, 5 – boundary of the park, 6 – boundary of protection zone. 80 g. fiedorowicz table 1 floristic-phytosociological differentiation of forest communities of the masurian landscape park successive no. 1 2 3 4 5 6 7 8 forest community p.-p. v.u.-p. s.-p. s.-p. t.-c. t.-c. t.-c. fr.-al. no. of the permanent research plot 1 2 6 9 4 8 10 7 forest complex krutyń krutyń kołoin gąsior krutyń kołoin gąsior kołoin forest section 101j 101c 245i 203f 53f 221i 215b 221g date 2.06. 1997 2.06. 1997 5.08. 1997 5.08. 1997 2.06. 1997 2.06. 1997 2.06. 1997 2.06. 1997 density of tree layer – a (%) 70 45 80 90 90 80 90 70 density of skrub layer – b (%) 20 10 5 50 5 5 5 5 cover of herb layer – c (%) 90 80 30 70 90 80 80 100 cover of moss layer – d (%) 90 95 90 10 10 5 5 10 surface of investigated plots in m2 400 400 400 400 400 400 400 400 number of plant species 26 23 30 44 34 35 30 27 ch. vaccinio-piceetea, dicrano-pinion pinus sylvestris l. a 4.4 2.2 1.1 4.4 . . 1.1 . pinus sylvestris l. b . 1.1 . . . . . . pinus sylvestris l. c + + + + . . . . picea abies (l.) h. karst. a 1.1 . 4.4 1.1 . . . . picea abies (l.) h. karst b 1.1 . + . . . . . picea abies (l.) h. karst c + + + + . . . . vaccinium myrtillus l. 4.5 1.2 1.1 1.1 . . . . pleurozium schreberi (willd. ex brid) mitt. d 4.4 +.2 4.4 1.1 . . . . ptilium crista-castrensis (hedw.) de not. d 1.2 . 2.2 . . . . . hylocomium splendens (hedw.) schimp. d 1.2 . 1.2 + . . . . dicranum polysetum sw. ex anon. d 1.2 . . . . . . . trientalis europaea l. + . 1.2 + . . . . vaccinium vitis-idaea l. 1.2 + + . . . . . lycopodium annotinum l. . + . . . . . . monotropa hypopitys l. s. s. . . . . + . . . ch. oxycocco-sphagnetea sphagnum magellanicum brid. d . 4.4 . . . . . . eriophorum vaginatum l. . 2.3 . . . . . . polytrichum strictum menzies ex brid. d . 1.2 . . . . . . sphagnum capillifolium (ehrh.) hedw. d . 1.1 . . . . . . oxycoccus palustris pers. . 1.2 . . . . . . andromeda polifolia l. . + . . . . . . drosera rotundifolia l. . + . . . . . . ch. et d. peucedano-pinetum peucedanum oreoselinum (l.) moench + . . . . . . . ch. et d. vaccinio uliginosi-pinetum vaccinium uliginosum l. . 3.3 . . . . . . ledum palustre l. . 2.3 . . . . . . ch. querco-fagetea, fagetalia sylvaticae acer platanoides l. a . . . . 1.1 1.1 . . acer platanoides l. c . . . + + + + . corylus avellana l. b . . 1.1 2.2 + 1.2 + + dryopteris filix-mas (l.) schott . . + + + + + . daphne mezereum l. . . . + . + . . ranunculus lanuginosus l. . . . + . . . + sanicula europaea l. . . . + . . . . anemone nemorosa l. . . . . 2.2 + 2.2 2.2 hepatica nobilis schreb. . . . . 1.1 1.2 + . mercurialis perennis l. . . . . 1.2 1.1 1.2 2.2 galium odoratum (l.) scop. . . . . 1.1 1.1 1.1 . galeobdolon luteum huds. . . . . 1.2 + + + asarum europaeum l. . . . . 1.2 + + . pulmonaria obscura dumort. . . . . + 1.1 + . lathyrus vernus (l.) bernh. . . . . + + 1.1 . poa nemoralis l. . . . . + + + . aegopodium podagraria l. . . . . + + + . macromycetes in selected forest communities 81 viola reichenbachiana jord. ex boreau . . . . + + + . atrichum undulatum (hedw.) p. beauv. d . . . . + + . . phyteuma spicatum l. . . . . + + . . polygonatum multiflorum (l.) all. . . . . + . . . neottia nidus-avis (l.) rich. . . . . + . . . lilium martagon l. . . . . + . . . lathraea squamaria l. . . . . . 1.2 + . corydalis solida (l.) clairv. . . . . . . 3.3 . anemone ranunculoides l. . . . . + . 1.1 3.3 fraxinus excelsior l. a . . . . . . . 3.3 ulmus glabra huds. a . . . . . . . 1.1 ulmus glabra huds. b . . . . . . . + stachys sylvatica l. . . . . . . 1.1 milium effusum l. . . . . . . . + ch. carpinion betuli tilia cordata mill. a . . . . 4.4 3.3 2.2 . tilia cordata mill. b . . . . 1.1 + 1.1 . tilia cordata mill. c . . . . + + + . carpinus betulus l. a . . . . 2.2 2.2 3.3 . carpinus betulus l. b . . . . + . + . carpinus betulus l. c . . . + + + 1.1 . stellaria holostea l. . . + + 1.1 1.1 1.1 . dactylis polygama horv. . . . . + + + . ch. alno-ulmion plagiomnium undulatum (hedw.) t. j. kop. d . . . . + + + 1.1 gagea lutea (l.) ker gawl. . . . . 1.1 . . + chrysosplenium alternifolium l. . . . . . . . 1.2 stellaria nemorum l. . . . . . . . + circaea lutetiana l. . . . . . . . + others betula pendula roth. a 1.1 . . . 1.1 1.1 + . quercus robur l. a . . . 2.2 1.1 1.1 1.1 + quercus robur l. b 1.1 . . 2.2 . . . . quercus robur l. c + + + + + . + . melampyrum pratense l. 1.2 + 1.1 + . . . . oxalis acetosella l. + . 2.2 3.3 . . . . maianthemum bifolium (l.) f.w. schmidt + . + + + . + . convallaria majalis l. . . + + . . + . plagiomnium affine (blandow ex funck) t. j. kop. d . + + + + + . mycelis muralis (l.) dumort. . . . + + + . + urtica dioica l. . . . + + + + + sporadic species: ajuga reptans l. 6 (+); alnus glutinosa (l.) gaertn. (a) 8 (2.2); a. glutinosa (l.) gaertn. (b) 8 (1.1); a. glutinosa (l.) gaertn. (c) 8 (+); anthoxanrhum odoratum l. 1 (+); athyrium filix-femina (l.) roth 8 (+); betula pendula roth. (b) 1 (+); b. pendula roth. (c) 1 (+), 3 (+); b. pubescens ehrh. (a) 2 (2.2); b. pubescens ehrh. (b) 2 (1.2); b. pubescens ehrh. (c) 2 (+); calamagrostis arundinacea (l.) roth 1 (+), 3 (1.1), 4 (+); calluna vulgaris l. 1 (+), 2 (+); campanula persicifolia l. 6 (+); c. rapunculoides l. 4 (+); cardamine flexuosa with. 8 (+); carex digitata l. 5 (+), 7 (+); dentaria bulbifera l. 6 (1.1); deschampsia caespitosa (l.) p. beauv. 1 (+), 3 (+), 4 (+); dicranum scoparium hedw. (d) 1 (+.2), 3 (+.2); digitalis grandiflora mill. 4 (+); dolichotheca seligeri (brid) loeske (d) 3 (1.2), 4 (1.1); dryopteris carthusiana (vill.) h. p. fuchs. 3 (+), 4 (+); dryopteris cristata (l.) a. gray 8 (+); erodium cicutarium (l.) l`hér. 6 (+); ficaria verna l. 8 (+); fragaria vesca l. 4 (+); frangula alnus mill. (b) 1 (+), 4 (+), 8 (+); galeopsis tetrahit l. 4 (+), 8 (+); galium mollugo l. 4 (+); geranium robertianum l. 4 (+); geum urbanum l. 6 (+); glechoma hederacea l. 8 (+); hieracium murorum l. 3 (+); impatiens parviflora dc. 4 (+), 8 (+); leucobryum glaucum (hedw.) angstr. (d) 1 (+.2); luzula pilosa (l.) willd. 3 (+), 4 (+); molinia caerulea (l.) moench 2 (+.2); polytrichum commune hedw. (d) 2 (1.2); polytrichum formosum (hedw.) g. l. sm. (d) 1 (+.2), 3 (1.2), 4 (+); prunella vulgaris l. 6 (+); pteridium aquilinum (l.) kuhn 3 (1.1), 4 (1.1); rubus ideus l. 3 (+), 4 (1.1); rumex acetosella l. 1 (+), 3 (+), 4 (+); sambucus racemosa l. (b) 4 (+); sorbus aucuparia l. em. hedl. (c) 1 (+), 3 (+), 4 (+); sphagnum cuspidatum ehrh. ex hoffm. (d) 2 (2.2); stellaria media (l.) vill. 6 (+); veronica officinalis l. 4 (+); viola canina l. 4 (+). explanations: p.-p. – peucedano-pinetum, v.u.-p. – vaccinio uliginosi – pinetum, s.-p. – serratulo-pinetum, t.-c. – tilio-carpinetum, fr.-al. – fraxino-alnetum. tab. 1. cont. 82 g. fiedorowicz table 2 macrofungi in the forest communities of the masurian landscape park successive no. 1 2 3 4 5 forest community p.-p. v.u.-p. s.-p. t.-c. fr.-al. total number of species 72 35 173 198 66 research plots pp cp pp cp pp cp pp cp pp cp number of plots 1 17 1 2 2 25 3 23 1 2 number of observations 39 19 39 2 78 31 117 52 39 2 mycorrhizal fungi number of species 49 19 75 50 9 paxillus involutus (batsch: fr.) fr. ss. lato 11+-2 7+-2 9+-2 12 26+-2 8+-2 38+-2 91-2 3+-1 . xerocomus badius (fr.: fr.) kühner ex gilbert 10+-1 8+-1 . . 24+-3 31-2 11 . . . amanita citrina (schaeff.) pers. 7+-2 6+-2 . . 9+-2 2+-1 11+-1 11 . . russula fragilis (pers.: fr.) fr. . 3+-1 . . 21 11 5+-1 21 . . cantharellus cibarius fr. 8+-1 41 . . 7+-2 41-2 4+-1 3+-1 . . russula vesca fr. 4+-1 1+ . . 12 2+-1 31-2 3+-1 . . russula cyanoxantha (schaeff.) fr. . 11 . . 7+-1 11 12+-1 11 . . boletus reticulatus schaeff. 2+-2 . . . . 21-2 11 2+-1 . . tricholoma terreum (schaeff.: fr.) p. kumm. ss. lato . 1+ . . 41 21 . 2+-1 . . lactarius necator (j. f. gmel.: fr.) pers. 11 11 . . 7+-2 3+-1 6+-1 21 . . russula velenovskyi melzer & zvára 3+ . . . 2+ . 1+ . . . amanita fulva (schaeff.) pers. . . 9+-2 1+ 4+-1 11 . 2+-1 . . lactarius rufus (scop.: fr.) fr. 15+-2 51-2 14+-4 21-2 30+-2 51-3 . . . . russula decolorans (fr.: fr.) fr. 14+-2 31-2 4+-1 1+ 6+-1 3+-1 . . . . laccaria proxima (boud.) pat. 8+-2 41-2 10+-1 . 8+-2 11 . . . . suillus variegatus (schwein.: fr.) o. kuntze 7+-2 11 6+-1 . . 1+ . . . . entoloma cetratum (fr.: fr.) m. m. moser 6+-1 21 51-2 . 10+-2 11 . . . . russula vinosa lindbl. 7+-2 11 . 1+ 7+-1 11 . . . . thelephora terrestris her. ex willd.: fr. . 11 6+-1 11 12+-2 2+-1 . . . . leccinum scabrum (bull.: fr.) gray 21 1+ 1+ 11 . 21 . . . . lactarius torminosus (schaeff.: fr.) pers. . 2+-1 . 11 . 11 . . . . lactarius vietus (fr.) fr. . 1+ . 11 . . . . . . cortinarius semisanguineus (fr.) gillet 14+-2 21-2 . . 5+-1 2+-1 . . . . cortinarius armillatus (fr.: fr.) fr. 9+-2 . . . 11 . . . . inocybe lacera (fr.: fr.) p. kumm. 9+-1 31 . . 8+-1 11 . . . . russula emetica (schaeff.) pers.: fr. var. silvestris 9+-1 4+-2 . . 4+-1 6+-2 . . . . cortinarius mucosus (bull.: fr.) kickx 7+-3 4+-2 . . 3+-1 11 . . . . amanita porphyria (alb. & schwein.: fr.) mladý 6+-2 21 . . 8+-2 4+-2 . . . . xerocomus subtomentosus (l.: fr.) quél. 4+-1 2+-1 . . 3+-1 21 . . . . tricholoma portentosum (fr.: fr.) quél. 5+-1 51-2 . . 3+-1 . . . . . tylopilus felleus (bull.: fr.) p. karst. 5+-1 2+-1 . . 9+-2 3+-1 . . . . suillus granulatus (l.: fr.) roussel 11 21-2 . . 8+-2 21 . . . . inocybe dulcamara (alb. & schwein.) p. kumm. . 11 . . 6+-1 31 . . . . lactarius mitissimus (fr.) fr. 2+-1 11 . . . 11 . . . . coltricia perennis (l.: fr.) murrill 11 11 . . . 21-2 . . . . suillus luteus (l.: fr.) roussel . 3+-2 . . 11 . . . . . russula sanguinea (bull.) fr. . 2+-1 . . . 1+ . . . . russula ochroleuca (pers.) fr. . 11 . . . 21 . . . . russula lutea (huds.: fr.) gray . 11 . . . 11 . . . . rozites caperatus (pers.: fr.) p. karst. 51-2 61-2 . . . . . . . . boletus pinophilus pilát & dermek 4+-1 3+-1 . . . . . . . . gomphidius roseus (fr.) fr. 2+-1 2+ . . . . . . . . suillus bovinus (l.: fr.) roussel 11 51-3 . . . . . . . . leccinum versipelle (fr.) snell 10+-2 . . . . . . . . . cortinarius cinnamomeus (l.: fr.) fr. 7+-2 . . . . . . . . . leccinum vulpinum watling 1+ . . . . . . . . . chalciporus piperatus (bull.: fr.) bat. . 31-2 . . . . . . . . tricholoma equeste (l.: fr.) p. kumm. ss. lato . 31-2 . . . . . . . . hygrophorus hypothejus (fr.: fr.) fr. . 11 . . . . . . . . russula versicolor jul. schäff. . 11 . . . . . . . . macromycetes in selected forest communities 83 cortinarius huronensis ammirati . . 20+-2 11 . . . . . . russula emetica (schaeff.) pers.: fr. var. emetica . . 11+-3 12 . . . . . . leccinum niveum (fr.) rauschert . . 7+-2 1+ . . . . . . russula paludosa britzelm. . . 6+-2 11 . . . . . . russula betularum hora . . 5+-1 1+ . . . . . . lactarius helvus (fr.) fr. . . 51-2 11 . . . . . . lactarius thejogalus (bull.: fr.) gray ss. neuhoff . . 3+-1 . . . . . . . laccaria laccata (scop.: fr.) berk. & broome . . . . 9+-2 11 40+-3 6+-2 21 . inocybe geophylla (fr.: fr.) p. kumm. . . . . 6+-2 11 18+-2 3+-1 6+-1 . laccaria amethystea (bull.) murrill . . . . 14+-2 4+-1 15+-2 41-2 11 . inocybe fastigiata (schaeff.) quél. . . . . 8+-2 . 9+-2 4+-1 7+-1 . lactarius pyrogalus (bull.: fr.) fr. . . . . 6+-1 11 13+-2 21 4+-1 . lactarius quietus (fr.) fr. . . . . 12+-1 11 23+-2 31-2 . . xerocomus pascuus (pers.) krombh. . . . . 10+-2 31 25+-3 51-3 . . russula puellaris fr. . . . . 6+-1 11 . 11 . . amanita pantherina (dc.: fr.) krombh. . . . . 6+-1 2+-1 3+-1 . . . tricholoma sulphureum (bull.: fr.) p. kumm. . . . . 4+-1 1+ 9+-1 21 . . russula pectinata (bull.) fr. ss. romagn. . . . . 3+-1 . 8+-1 1+ . . lactarius camphoratus fr. . . . . 21 11 . 5+-2 . . amanita muscaria (l.: fr.) hook. . . . . . 11 1+ 1+ . . amanita rubescens (pers.: fr.) gray . . . . 11 3+-1 14+-1 3+-1 . . craterellus cornucopiodes (l.: fr.) pers. . . . . . 12 . 42-4 . . scleroderma verrucosum (bull.): pers. . . . . 2+-2 . 1+ . . . russula xerampelina (schaeff.) fr. . . . . 7+-1 2+-1 . . . . boletus edulis bull.: fr. . . . . 3+-1 4+-1 . . . . chroogomphus rutilus (schaeff.: fr.) o. k. miller . . . . 21 2+-1 . . . . russula queletii fr. . . . . 21-2 1+ . . . . lactarius deterrimus gröger . . . . 12 12 . . . . gomphidius glutinosus (schaeff.: fr.) fr. . . . . 1+ 11 . . . . russula mustelina fr. . . . . 5+-1 . . . . . lactarius deliciosus (l.: fr.) gray . . . . 21 . . . . . amanita gemmata (fr.) bertillon . . . . 1+ . . . . . lactarius piperatus (l.: fr.) gray . . . . . 1+ 10+-1 3+-2 . . gyroporus castaneus (bull.: fr.) quél. . . . . . 2+-1 5+-1 1+ . . scleroderma citrinum pers. . . . . . 11 1+ 11 . . tricholoma saponaceum (fr.: fr.) p. kumm. . . . . . 21 . 2+-1 . . cortinarius brunneus (pers.: fr.) fr. . . . . . 2+-1 . . . . cortinarius traganus (fr.: fr.) fr. . . . . . 21 . . . . hygrophoropsis aurantiaca (wulf.: fr.) j. schröt. . . . . . 21 . . . . elaphomyces granulatus fr. . . . . . 12 . . . . lactarius volemus (fr.) fr. . . . . . 1+ . . . . russula badia quél. . . . . . 11 . . . . russula foetens (pers.: fr.) fr. . . . . . 1+ . . . . suillus grevillei (klotzsch: fr.) singer . . . . . 12 . . . . hydnum repandum l.: fr. . . . . . . 13+-2 6+-2 . . entoloma rhodopolium for. nidorosum (fr.) noordel. . . . . . . 13+-1 41 . . tricholoma lascivum (fr.: fr.) gillet . . . . . . 13+-1 2+-1 . . russula nigricans (bull.: fr.) fr. . . . . . . 101-2 21 . . russula risigallina (batsch) sacc. . . . . . . 9+-1 21-2 . . amanita phalloides (vaill.: fr.) link . . . . . . 7+-1 5+-2 . . hebeloma crustuliniforme (bull.) quél. . . . . . . 7+-2 2+-1 . . leccinum pseudoscabrum (kallenb.) šutara . . . . . . 5+-2 8+-2 . . lactarius vellereus (fr.) fr. . . . . . . 3+-1 11 . . russula delica fr. . . . . . . 8+-2 . . . amanita vaginata (bull.: fr.) vittad . . . . . . 11 . . . boletus luridus schaeff.: fr. . . . . . . . 11 . . cortinarius orellanus fr. . . . . . . . 11 . . inocybe erubescens blytt . . . . . . . 11 . . leccinum aurantiacum (bull.) gray . . . . . . . 12 . . tab. 2. cont. 84 g. fiedorowicz pseudocraterellus undulatus (pers.: fr.) rauschert . . . . . . . 14 . . russula solaris ferd. & winge . . . . . . . 11 . . xerocomus rubellus (krombh.) quél. . . . . . . . 11 . . cortinarius paleaceus fr. . . . . . . . . 4+-1 . lactarius lilacinus (lasch: fr.) fr. . . . . . . . . 3+-1 . paxillus rubicundulus p. d. ordon . . . . . . . . 1+ . saprotrophic fungi on humus number of species 0 0 13 36 8 lycoperdon perlatum pers.: pers. . . . . . 11 20+-2 31 51-2 . humaria hemisphaerica (f. h. wigg.: fr.) fuckel . . . . 5+-1 . 18+-3 7+-2 . . cystolepiota seminuda (lasch) bon . . . . 3+-1 1+ 27+-2 4+-1 5+-1 . agaricus silvicola (vittad.) peck . . . . 3 +-1 1+ 4+-1 . . . lycoperdon umbrinum pers.: pers. . . . . 141-2 4+-2 . . . . otidea leporina (batsch: fr.) fuckel . . . . 31-2 12 . . . . agaricus silvaticus schaeff. . . . . 3+-1 . . . . . peziza badia pers.: fr. . . . . . 3+-1 . . . . geoglossum umbratile sacc. . . . . . 13 . . . . gyromitra esculenta (pers.) fr. . . . . . 11 . . . . lyophyllum decastes (fr.: fr.) singer . . . . . 12 . . . . macrolepiota rhacodes (vittad.) singer . . . . . 11 . . . . morchella conica pers. . . . . . 11 . . . . phallus impudicus l.: pers. . . . . . . 19+-2 41-2 2+ . lepiota cristata (bolt.: fr.) p. kumm. . . . . . . 6+-1 41-2 5+-1 . gyromitra gigas (krombh.) cooke . . . . . . 2+-1 21-2 1+ . clavulina coralloides (l.: fr.) j. schröt. . . . . . . 24+-2 6+-1 . . bovista nigrescens pers.: pers. . . . . . . 16+-2 . . . macrolepiota procera (scop.: fr.) singer . . . . . . 14+-2 21 . . calvatia excipuliformis (scop.: pers.) perdeck . . . . . . 10+-1 21 . . clavulina cinerea (bull.: fr.) j. schröt. . . . . . . 6+-1 21 . . calocybe gambosa (fr.) donk . . . . . . 3+-2 11 . . helvella macropus (pers.) p. karst. . . . . . . 3+-1 11 . . lepiota subgracilis kühner ex wasser . . . . . . 3+ . . . peziza arvernensis boud. . . . . . . 3+-2 . . . otidea onotica (pers.: fr.) fuckel . . . . . . 21-2 32-3 . . coprinus xanthothrix romagn. . . . . . . 2+-1 11 . . peziza succosa berk. . . . . . . 21 . . . peziza vesiculosa bull. . . . . . . 2+-1 . . . clavariadelphus pistillaris (l.: fr.) donk . . . . . . 1+ . . . agaricus placomyces peck . . . . . . 11 11 . . lycoperdon molle pers. . . . . . . 1+ . . . helvella crispa (scop.) fr. . . . . . . . 61-2 . . scutellinia scutellata (l.) lambotte . . . . . . . 61-2 . . coprinus atramentarius (bull.: fr.) fr. . . . . . . . 21-2 . . helvella elastica bull. . . . . . . . 21 . . thelephora palmata (scop.): fr. . . . . . . . 21 . . albatrellus confluens (fr.) kotl. & pouzar . . . . . . . 12 . . aleuria aurantia (pers.: fr.) fuckel . . . . . . . 14 . . calvatia utriformis (bull.: pers.) jaap . . . . . . . 11 . . helvella acetabulum (l.: fr.) quél. . . . . . . . 12 . . langermannia gigantea (batsch: pers.) rostk. . . . . . . . 1+ . . lyophyllum connatum (schum.: fr.) singer . . . . . . . 12 . . morchella esculenta (l.) pers. . . . . . . . 1+ . . otidea cochleata (l.) fuckel . . . . . . . 12 . . conocybe tenera (schaeff.: fr.) fayod . . . . . . . . 8+-2 . ramaria aurea (schaeff.: fr.) quél. . . . . . . . . 81-2 . mutinus caninus (huds.: pers.) fr. . . . . . . . . 3+-1 . saprotrophic fungi on litter number of species 9 4 27 21 11 mycena galopus (pers.: fr.) p. kumm. 11+-2 7+-2 13+-2 11 32+-3 8+-2 9+-1 4+-1 11+-2 . setulipes androsaceus (l.: fr.) antonín 131-5 61-4 22+-4 21-2 23+-4 31-2 . . . . tab. 2. cont. macromycetes in selected forest communities 85 mycena epipterygia (scop.: fr.) gray 8+-2 41-2 10+-3 11 7+-2 31-2 . . . . rhodocollybia butyracea for. asema (fr.: fr.) antonín, halling & noordel 8+-1 41-2 . . 20+-4 31-2 . . . . cystoderma amianthinum (scop.: fr.) fayod 7+-2 4+-1 . . 11+-2 11 . . . . strobilurus tenacellus (pers.: fr.) singer . 11 6+-1 . 101-2 . . . . . gymnopus dryophilus (bull.: fr.) murrill . 21 . . 16+-2 12 38+-2 71-3 12+-2 11 mycena sanguinolenta (alb. & schwein.: fr.) p. kumm. . 12 . . 10+-1 31-2 101-2 41-2 31-2 . auriscalpium vulgare gray . 31 . . 22+-2 11+-2 31 . . . mycena pura (pers.: fr.) p. kumm. . . . . 17+-2 41 33+-2 41-2 10+-2 . mycena vitilis (fr.) quél. . . . . 6+-2 . 40+-2 6+-2 3+-1 . clitocybe gibba (pers.: fr.) p. kumm. . . . . 9+-2 1+ 27+-2 21 . . hymenoscyphus fructigenus (bull.) fr. . . . . 11 . 71-2 21-2 . . psilocybe aeruginosa (m. a. curtis: fr.) noordel. . . . . . 21 21+-1 12 . . mycena stylobates (pers.: fr.) p. kumm. . . . . 61-2 . 18+-2 51-2 . . gymnopus confluens (pers.: fr.) antonín, halling & noordel. . . . . 51-2 12 32 13 . . gymnopus peronatus (bolt.: fr.) antonín, halling & noordel. . . . . 4+-2 2+-3 31 . . . clitocybe clavipes (pers.: fr.) p. kumm. . . . . 19+-2 4+-1 . . . . mycena zephirus (fr.: fr.) p. kumm. . . . . 9+-2 41-2 . . . . strobilurus esculentus (wulf.: fr.) singer . . . . 8+-4 . . . . . mycena aetitis (fr.) quél. . . . . 7+-1 . . . . . ramaria abietina (pers.: fr.) quél. . . . . 61-2 11 . . . . marasmius scorodonius (fr.: fr.) fr. . . . . 2+-2 12 . . . . marasmiellus perforans (hoffm.: fr.) antonín, halling & noordel. . . . . 21-2 12 . . . . rutstroemia bulgarioides (rabenh.) p. karst. . . . . . 12 . . . . strobilurus stephanocytis (hora) singer . . . . . 11 . . . . cystodermella granulosum (batsch: fr.) harmaja . . . . . 11 . . . . marasmius rotula (scop.: fr.) fr. . . . . . . 44+-4 91-3 181-4 . rhodocollybia butyracea (bull.: fr.) lennox . . . . . . 26+-2 6+-2 21 . clitocybe nebularis (batsch: fr.) p. kumm. . . . . . . 8+-2 31 6+-1 . tubaria furfuracea (pers.: fr.) gillet . . . . . . 9+-3 31-2 41-2 . mycena polyadelpha (lasch) kühner . . . . . . 131-4 51-3 . . marasmiellus foetidus (sowerby: fr.) antonín, halling & noordel. . . . . . . . 14 . . clitocybe geotropa (bull.) quél. . . . . . . 5+-1 21 . . setulipess quercophilus (pouzar) antonín . . . . . . 61-2 22 . . lepista nuda (bull.: fr.) cooke . . . . . . . 21 . . hymenoscyphus albidus (roberge ex desm.) w. phillips . . . . . . . . 133-5 . mycena rubromarginata (fr.: fr.) p. kumm. . . . . . . . . 31-2 . saprotrophic fungi on wood number of species 10 4 41 76 30 stereum hirsutum (willd.: fr.) gray x2 11 . . x2 81-2 3x2 271-3 x3 12 exidia plana (wiggers) donk 111-2 22 . . 121-2 42 351-3 102-3 . . pluteus atricapillus (batsch) fayod . . 1+ . 5+-1 4+-1 16+-2 3+-1 4+-1 . piptoporus betulinus (bull.: fr.) p. karst. . . 51 . . . 10+-1 31 . . polyporus brumalis (pers.): fr. . 1+ . . . 1+ 31 . . . calocera viscosa (pers.: fr.) fr. 5+-1 11 . . 5+-1 31-2 . . . . xeromphalia campanella (batsch: fr.) kühner & maire 51-3 12 . . . 41-4 . . . . psilocybe capnoides (fr.: fr.) noordel. 21-2 21-4 . . 42 32-3 . . . . gymnopilus penetrans (fr.: fr.) murrill . 21 . . 71-2 31-2 . . . . lentinus lepideus (fr.: fr.) fr. . 12 . . 12 . . . . . trichaptum abietinum (dicks.: fr.) ryvarden . 31-2 . 11 x2 12 . . . . trichaptum fuscoviolaceum (ehrenb.: fr.) ryvarden . 12 . . 2x2 41-2 . . . . rhodocollybia maculata (alb. & schwein.: fr.) singer . . 11 . . . . . . . tab. 2. cont. 86 g. fiedorowicz stereum rugosum (pers.: fr.) fr. . . . . x1-3 . 2x1-3 82 x3 12 mycena galericulata (scop.: fr.) gray . . . . 5+-1 12 32+-2 31-2 141-2 . psilocybe lateritia (schaeff.: fr.) noordel. . . . . 12 . 12 11 12 . psilocybe fasticularis (huds.: fr.) noordel. . . . . 101-3 22 162-4 62-4 12 . ramaria stricta (pers.: fr.) quél. . . . . . 11 3+-1 31-2 . 11 xylaria hypoxylon (l.) grev. . . . . . 41-2 3x1-4 282-3 x3 12 xylaria polymorpha (pers.) grev. . . . . . 22 3x1-3 251-3 x1 12 tremella mesenterica retz.: fr. . . . . . 11 21 . . . daedalea quercina (l.: fr.) pers. . . . . . 11 . 11 . . exidia glandulosa (bull.): fr. . . . . 61-2 . 51-2 31-2 . . hyphodontia paradoxa (schrad.: fr.) e. langer & vesterholt ss. lato . . . . x2 . 3x3-4 132-4 . . pholiota mutabilis (scop.: fr.) p. kumm. . . . . 71-2 21-2 181-2 42 . . crepidotus variabilis (pers.: fr.) p. kumm. . . . . . 31-2 25+-2 31-2 . . trametes versicolor (l.: fr.) pilát . . . . . 12 2x2 21-2 . . geopyxis carbonaria (alb. & schwein.: fr.) sacc. . . . . . 11 . . . . lycoperdon pyriforme schaeff.: pers. . . . . 17+-2 21-2 18+-2 41-2 . . mycena stipata maas geest. & schwöb. . . . . 81-2 . 51-2 31-2 . . stereum sanguinolentum alb. & schwein.: fr.) fr. . . . . 311-3 22 . . . . pseudohydnum galatinosum (scop.: fr.) p. karst. . . . . 5+-1 . . . . . pluteus atromarginatus (singer) kühner . . . . 3+-1 . . . . . tricholomopsis rutilans (schaeff.: fr.) singer . . . . 2+-1 11 . . . . discina ancilis (pers.) sacc. . . . . . 12 . . . . oligoporus stipticus (pers.: fr.) gilbertson & ryvarden . . . . . 4+-1 . . . . panellus mitis (pers.: fr.) singer . . . . . 32 . . . . gloephyllum odoratum (wulf.: fr.) imaz. . . . . . 21 . . . . gloephyllum sepiarum (wulf.: fr.) p. karst. . . . . . 11 . . . . hymenochaete tabacina (sowerby) lév. . . . . . 12 . . . . phaeolus schweinitzii (fr.: fr.) pat. . . . . . 1+ . . . . phlebia tremellosa (schrad.: fr.) nakasone & burds. . . . . . 11 . . . . oligoporus caesius (schrad.: fr.) gilbertson & ryvarden . . . . . 11 . . . . ganoderma applanatum (pers.) pat. . . . . . . 2x1-2 11 . 11 hymenochaete rubiginosa (schrad.: fr.) lév. . . . . . . x2 12 . . megacollybia platyphylla (pers.: fr.) kotl. & pouzar . . . . . . 18+-2 5+-1 10+-2 . polyporus ciliatus fr.: fr. . . . . . . 121-4 32-3 61 12 mycena inclinata (fr.) quél. . . . . . . 92-4 . 72-5 . panellus serotinus (schrad.: fr.) kühner . . . . . . 92-3 . 42 . polyporus badius (pers.) schwein. . . . . . . 72-3 . . 1+ sarcoscypha austriaca (o. beck ex sacc.) boud. . . . . . . 71-4 41-2 6+-3 12 delicatula integrella (pers.: fr.) fayod . . . . . . 22 12 21-2 . datronia mollis (sommerf.: fr.) donk . . . . . . . 81-2 x2 . polyporus melanopus (pers.): fr. . . . . . . 11 . 11 . coprinus disseminatus (pers.: fr.) quél. . . . . . . . 32-3 23 . polyporus squamosus (huds.): fr. . . . . . . . 21-2 51 . bjerkandera adusta (willd.: fr.) p. karst. . . . . . . . 41-2 . 12 discina parma . breitenb. & maas geest. . . . . . . . 12 . 11 trametes hirsuta (wulf.: fr.) pilát . . . . . . 2x2 51-2 . . peniophora quercina (pers.: fr.) cooke . . . . . . 2x2-3 52 . . xylaria longipes nitschke . . . . . . 2x1-2 101-2 . . cylindrobasidium laeve (pers.: fr.) chamuris . . . . . . 2x2 72 . . nectria cinnabarina (tode: fr.) fr. . . . . . . x3 42-3 . . peniophora incarnata (pers.: fr.) p. karst. . . . . . . x2 42 . . peniophora rufomarginata (pers.) litsch. . . . . . . x2 22 . . mycena polygramma (bull.: fr.) gray . . . . . . 21+-2 41-2 . . mycena maculata p. karst. . . . . . . 181-3 42 . . phlebia radiata fr. . . . . . . 141-3 51-2 . . crepidotus mollis (schaeff.: fr.) staude . . . . . . 131-2 51-2 . . tab. 2. cont. macromycetes in selected forest communities 87 cyathus striatus (huds.) willd.: pers. . . . . . . 91-4 32 . . xerula radicata (relh.: fr.) dörfelt . . . . . . 8+-2 . . . schizophyllum commune fr.: fr. . . . . . . 72 42 . . pleurotus pulmonarius (fr.) quél. . . . . . . 7+-2 31-2 . . psathyrella candolleana (fr.: fr.) maire . . . . . . 52-3 . . . polyporus varius (pers.): fr. . . . . . . 5+-1 3+-1 . . polyporus arcularius (batsch): fr. . . . . . . 4+-1 . encoelia furfuracea (roth) p. karst. . . . . . . 32 . . . pluteus salicinus (pers.: fr.) p. kumm. . . . . . . 2+-1 . tremella foliacea pers. . . . . . . 21 21 . . bisporella citrina (batsch: fr.) korf & s. e. carp. . . . . . . 22 52-3 clavariadelphus fistulosus (holmsk.: fr.) coener . . . . . . 21 . . . genoderma lucidum (m. a. curtis: fr.) p. karst. . . . . . . 2+ . . . pleurotus ostreatus (jacq.: fr.) p. kumm. . . . . . . 11 21-2 . . lentinellus cochleatus (pers.: fr.) p. karst. . . . . . . 12 12 . . crucibulum leave (huds.) kambly . . . . . . 11 . . . paxillus atrotomentosus (batsch: fr.) fr. . . . . . . 1+ . . . xerula pudens (pers.) singer . . . . . . 12 . . . bjerkandera fumosa (pers.: fr.) p. karst. . . . . . . . 82 . . panellus stypticus (bull.: fr.) p. karst. . . . . . . . 61-3 . . calocera cornea (batsch: fr.) fr. . . . . . . . 41-2 . . ascocoryne sarcoides (jacq.) j. w. groves & d. e. wilson . . . . . . . 42 . . cerrena unicolor (bull.: fr.) murrill . . . . . . . 42 . . flammulina velutipes (m. a. curtis: fr.) singer . . . . . . . 32-3 . . bulgaria inquinalis (pers.) fr. . . . . . . . 22 . . lenzites betulinus (l.: fr.) fr. . . . . . . . 22 . . coprinus micaceus (bull.: fr.) fr. . . . . . . . 23-4 . . pleurotus dryinus (pers.: fr.) p. kumm. . . . . . . . 1+ . . pycnoporus cinnabarinus (jacq.: fr.) p. karst. . . . . . . . 11 . . peniophora limitata (chaillet: fr.) cooke . . . . . . . . x2 12 chlorociboria aeruginosa (pers.: fr.) seaver ex c. s. ramamuthri, korf & l. r. batra . . . . . . . . 2+-1 . daedaleopsis confragosa (bolt.: fr.) j. schröt. . . . . . . . . x1 11 stereum subtomentosum pouzar . . . . . . . . x2 . mycena niveipes murrill 41-2 pluteus nanus (pers.: fr.) p. kumm. . . . . . . . . 1+ . hypoxylon fuscum (pers.: fr.) fr. . . . . . . . . . 12 bryophilous fungi number of species 2 5 5 4 3 galerina hypnorum (schrank: fr.) kühner 21 11 . . 27+-2 21 11+-2 4+-1 101-2 . rickenella fibula (bull.: fr.) raith. 21 . . . 71-2 11 16+-2 21-2 9+-3 . mycena acicula (schaeff.) p. kumm. . . . . 9+-1 . 17+-2 21 . . galerina paludosa (fr.) kühner . . 14+-2 21-2 . . . . . . galerina sphagnorum (pers.: fr.) kühner . . 13+-2 . . . . . . . psilocybe elongata (pers.: fr.) j. e. lange . . 10+-2 11 . . . . . . lyophyllum palustre (peck) singer . . 15+-2 1+ . . . . . . omphalina sphagnicola (berk.) m. m. moser . . . 1+ . . . . . . galerina mniophila (lasch) kühner . . . . 9+-1 21 . . . . leotia lubrica (scop.) pers. . . . . . 21-2 . . . . rickenella setipes (fr.: fr.) raith. . . . . . . 9+-1 21 7+-1 . parasitic fungi number of species 3 3 10 11 5 fomes fomentarius (l.: fr.) kickx . . x1 . . 21 2x1 131-2 . 11 fomitopsis pinicola (swartz: fr.) p. karst. . . 291 1+ x1 21 2x1-2 11 . . sparassis crispa (wulf.): fr. 2+ . . . 1+ 2+-1 . . . . heterobasidion annosum (fr.) bref. ss. lato . 2+-1 . . x1-2 . . . . . inonotus obliquus (pers.: fr.) pilát . 11 . 1+ . . . . . . armillaria ostoyae (romagn.) herink . . . . 21 21-2 52 41-2 21-2 . laetiporus sulphureus (bull.: fr.) murrill . . . . . 5+-1 41-2 2+-1 . . tab. 2. cont. 88 g. fiedorowicz three hundred thirty five taxa of macromycetes were recorded in the phytocoenoses studied. a total of 354 macromycete species were observed between 1997 and 2000. the total number of taxa of macromycete species, including literature data, reported from the area is 506. peucedano-pinetum (w. mat. 1962) w. mat. & j. mat. 1973 mycological observations in peucedano-pinetum were conducted in one permanent plot (no. 1) and at 17 supplementary plots. a total of 58 observations in communities classified as peucedano-pinetum were performed. the occurrence of 73 macromycete species was recorded in the association (tab. 2). eleven macromycete species were exclusive of the association in the mlp, which is 15.3% of the mycobiota in peucedano-pinetum. those are: boletus pinophilus, chalciporus piperatus, cortinarius cinnamoneus, gomphidius roseus, hygrophorus hypothejus, leccinum versipelle, l. vulpinum, rozites caperatus, suillus bovinus, russula versicolor and tricholoma equestre. the highest number of species in common was recorded between peucedano-pinetum and serratulo-pinetum (60 species, over 83%). peucedano-pinetum is an association characteristic of northeastern poland. it is also known as vaccinio myrtilli-pinetum kobendza 1930 (matuszkiewicz 2001). ma cromycetes occurring in peucedano-pinetum, mostly from the białowieża old growth forest – project crypto, are reported only sporadically in the available lite rature (nespiak 1959; chmiel, sadowska 1994; lisiewska 1995; bujakiewicz 1995; chmiel 1995, 1996; skirgiełło 1995). mycological observations in peucedano-pinetum outside the białowieża old growth forest have been conducted by, for instance, lisiewska (1991-1992) and łuszczyński (2007). data on the mycobiota of leucobryo-pinetum, which is vicariant with peucedano-pinetum and which occurs in other parts of poland, are reported in mycological literature more often (e.g., lisiewska 1978; lisiewska, wójcik 1984; friedrich 1985, 1994; ławrynowicz, szkodzik 1998; łuszczyński 2007). the mycobiota of the peucedano-pinetum phytocoenoses examined in the mlp corresponds to literature data; this is probably related to the characteristic species composition of peucedano-pinetum and its vicariant leucobryo-pinetum. macromycete species associated with pinus sylvestris dominate in both types of coniferous forests. tremella encephala pers.: fr. . . . . 13+-2 21 . . . . phellinus pini (brot.: fr.) a. ames . . . . . 11 . . . . cordyceps capitata (holmskj.: fr.) fr. . . . . . 12 . . . . cordyceps ophioglossoides (ehrh. ex pers.: fr.) fr. . . . . . 11 . . . . pholiota squarrosa (weigel: fr.) p. kumm. . . . . . . 51 . 32-4 . dumontinia tuberosa (hedw.) l. m. kohn . . . . . . 22 11 41-2 11 inonotus radiatus (sowerby: fr.) p. karst. . . . . . . . 13 . 12 fistulina hepatica (schaeff.): fr. . . . . . . 8+-1 2+-1 . . phellinus robustus (p. karst.) bourdot & galzin . . . . . . x1 11 . . phellinus alni (bondartsev) parmasto . . . . . . . 11 . . asterophora parasitica (pers.: fr.) singer . . . . . . . 12 . . abbreviations: p.-p. – peucedano-pinetum, v.u.-p. – vaccinio uliginosi – pinetum, s.-p. – serratulo-pinetum, t.-c. – tilio-carpinetum, fr.-al. – fraxino-alnetum; pp – permanent plots, cp – complementary plots; 1, 2, 3... – the number of records at a given plot; x – which corresponds to the sum of all observations at a specific plot, is used for species producing permanent fruit-bodies; +-5 – the commonly used moser scale in the quantitative assessment. tab. 2. cont. macromycetes in selected forest communities 89 vaccinio uliginosi-pinetum kleist 1929 observations in vaccinio uliginosi-pinetum were conducted in one permanent plot (no. 2) and 2 supplementary plots. a total of 41 observations were performed (tab. 2). the occurrence of 35 macromycete species, including 13 (37.1%) species exclusive of the community, was recorded in vaccinio uliginosi-pinetum. species that occur only in vaccinio uliginosi-pinetum are as follows: cortinarius huronensis, galerina paludosa, g. sphagnorum, lactarius helvus, l. thejogalus, leccinum niveum, lyophyllum palustre, omphalina sphagnicola, psilocybe elongata, russula betularum, r. emetica var. emetica, r. paludosa and rhodocollybia maculata. the greatest resemblance of the mycobiota in the association was observed in serratulo-pinetum (19 species in common between both associations – 54.3% of the biota) and peucedano-pinetum (17 species in common between both associations – 48.6% of the biota). it is understandable given the coniferous character of the association and the dominance of pinus sylvestris. the mycobiota of vaccinio uliginosi-pinetum has been examined in a few mycocoenological studies (e.g. nespiak 1959; bujakiewicz 1986; lisiewska 1978; kałucka 1995; friedrich 1997; łuszczyński 2007). the majority of macromycete species recorded in the association have also occurred in patches of vaccinio uliginosi-pinetum in other parts of poland. those were, for instance, cortinarius huronensis, galerina paludosa, g. sphagnorum, lactarius helvus, leccinum niveum, lyophyllum palustre, psilocybe elongata, russula emetica var. emetica, r. paludosa (bujakiewicz 1986; kałucka 1995). serratulo-pinetum (w. mat. 1981) j. mat. 1988 macromycetes in serratulo-pinetum were examined in 2 permanent plots (nos. 6 and 9) and 25 supplementary plots (tab. 2). a total of 109 observations were performed in the association. the occurrence of 171 macromycete species was recorded. fifty six species exclusive of the association were found (32.4% of the mycobiota of the association). those included: agaricus silvaticus, boletus edulis, chroogomphus rutilus, clitocybe clavipes, lactarius deterrimus, lycoperdon umbrinum, macrolepiota rhacodes, mycena zephirus, russula mustelina, r. xerampelina, stereum sanguinolentum or strobilurus esculentus. the mixed tree-stand affects the number of species in common between serratulo-pinetum phytocoenoses and other plant associations. the greatest number of species in common was recorded in tilio-carpinetum – 76 species (43.9%) and peucedano-pinetum – 60 species (34.7%). this is characteristic of a community that exhibits features of both coniferous forests and oak-hornbeam forests and was observed in both permanent plots. the plot with a distinct participation of picea abies (plot 6, tab. 1) exhibits more features of the coniferous forest, which corresponds to fungal species observed in the plot (e.g., pseudohydnum gelatinosum, ramaria abietina, russula mustelina, strobilurus esculentus, etc.). a considerable participation of quercus robur in plot 9 results in its greater affiliation with broadleaved communities, which is reflected in the species composition of macromycetes (e.g., cystolepiota seminuda, tricholoma sulphureum, hyphodontia paradoxa, etc.). similarly to peucedano-pinetum, the mycobiota of serratulo-pinetum is one of the most mycocoenologically under-explored mycobiotas in poland. the subboreal mixed coniferous forest occurs in northeastern poland and sporadically in central and eastern poland. it is known as pino-quercetum serratuletosum or calamagrostio 90 g. fiedorowicz arundinaceae-piceetum in older studies from northeastern poland (matuszkiewicz 2001). data on the mycobiota of serratulo-pinetum are available in studies by nespiak (1959), lisiewska (1991-1992) and łuszczyński (2007). the mycobiota in the serratulo-pinetum phytocoenosis is intermediate between coniferous communities and oak-hornbeam communities. species characteristic of both coniferous communities and broadleaved communities occurred there. tilio-carpinetum tracz. 1962 macromycetes of oak-hornbeam forests represented by tilio-carpinetum were observed in 3 permanent plots (nos. 4, 8, 10; tab. 1) and 23 supplementary plots in the mlp. a total of 169 observations were performed and a total of 198 macromycete species were recorded. no considerable differences in the species composition among individual permanent plots were observed. the mycobiota of tilio-carpinetum was characterised by the highest participation of exclusive species – 78 ones, that is 39.4% of the mycobiota of the association. clavulina coralloides, cylindrobasidium laeve, mycena maculata, peniophora quercinum, p. rufomarginata, phlebia radiata, phellinus robustus, xylaria longipes were the most frequently recorded species exclusive to oak-hornbeam forests in the mlp. species exclusive of the oak-hornbeam forest also included: amanita phalloides, clavariadelphus fistulosus, entoloma nidorosum, fistulina hepatica, leccinum pseudoscabrum, macrolepiota procera, pleurotus pulmonarius, tricholoma lascivum, russula risigallina, xerula pudens. the greatest similarity was observed between the mycobiota of the oak-hornbeam forest and fungi in serratulo-pinetum (76 species occurring in both; 38.4% of the mycobiota in the association). the number of species that occurred in tiliocarpinetum and other broadleaved communities was at a similar level: slightly over 20% of the mycobiota in the association. the species composition of macromycetes in tilio-carpinetum has been studied intensely in poland (e.g., nespiak 1959; ławrynowicz 1973; lisiewska 1978; gumińska 1991-1992; skirgiełło 1998; wojewoda et al. 1999; ławrynowicz et al. 2002, łuszczyński 2007). the mycobiota of the association examined in this study differentiates it well from other forest communities investigated in the mlp. the species composition of macromycetes in tilio-carpinetum is mostly consistent with fungal species occurring in oak-hornbeam phytocoenoses in other parts of poland. the analysis shows that the greatest similarity of the species composition and the number of species is observed between the local mycobiota of tilio-carpinetum in the mlp and the mycobiotas in tilio-carpinetum recorded in central poland (ławrynowicz 1973) and the skołczanka reserve (gumińska 1991-1992) and in the tuchola forests (ławrynowicz et al. 2002). fraxino-alnetum w. mat. 1952 the mycobiota of fraxino-alnetum was examined in one permanent plot (no. 7; tab. 1) and 2 supplementary plots. a total of 41 mycological observations were performed. 66 species of macromycetes were recorded (tab. 2). exclusive species (ten species) constituted 15.2% of the association mycobiota. cortinarius paleaceus, daedaleopsis confragosa, lactarius lilacinus, mutinus caninus, mycena niveipes, m. rubromarginata, paxillus rubicundulus, pluteus nanus, ramaria aurea and stereum subtomentosum occurred only in fraxino-alnetum in the mlp. fraxino-alnetum and tilio-carpinetum had the greatest number of species in common (46 species, 69.7% of the association biota). 92 g. fiedorowicz ratio was observed for saprotrophic fungi on wood which was only 11.4% for vaccinio uliginosi-pinetum and 13.9% for peucedano-pinetum in coniferous associations. their greatest values were observed for the fraxino-alnetum phytocoenosis (45.4% for fraxino-alnetum). the ratio between the number of macromycete species and the number of plants in permanent plots in the phytocoenoses examined ranges from 1:1.81 for vaccinio uliginosi-pinetum to 3.79:1 for tilio-carpinetum (tab. 3). the data are comparable with the results obtained by łuszczyński (2007). however, the ratio is 1:1.81 (506 macromycete species : 920 plant species) for the total number of macromycete species recorded in the mlp (the author’s observations and literature data) and the number of plant species (kruszelnicki 1996). however, the observations by mułenko (1998) and grzywacz (1999), revealed that the number of fungal species is much greater than the number of plant species when macromycetes are also included. summary three hundred and thirty five taxa of macromycetes were recorded in the phytocoenoses examined in the masurian landscape park; 354 macromycete species were observed during examinations conducted in the park between 1997 and 2000. the total number of species reported from the masurian landscape park, including literature data, is 506. tilio-carpinetum (198 taxa) and serratulo-pinetum (171 taxa) were the richest phytocoenoses in macromycete species. this is confirmed by the relationship between the number of macromycete species occurring in them and the diversified tree-stand (the diversity of substrate available for mycelium development). seventy three taxa of macromycetes were recorded in peucedano-pinetum. the association in the study area is differentiated by a group of exclusive species (15.5% of the mycobiota of the association) such as boletus pinophilus, cortinarius cinnamoneus, rozites caperatus, russula versicolor or tricholoma equestre as well as a group of species that it has in common with the serratulo-pinetum phytocoenosis (83% of the mycobiota of the association). the species composition of macromycetes in serratulo-pinetum is characterised by a high participation of species in common with broadleaved communities (47.9% of the mycobiota in common with tilio-carpinetum) and with coniferous forests (34.7% of the mycobiota in common with peucedano-pinetum). 32.4% of exclusive species was recorded in the continental mixed coniferous forest. a downward trend in the percentage participation of mycorrhizal species from coniferous communities (43-66% of the mycobiota of the associations) to broadleaved communities (12-27% of the mycobiota) was observed. a reverse trend was noted in the case of saprotrophic fungi on wood. the greatest participation of this group was recorded in tilio-carpinetum (ca 35%) and in fraxino-alentum (45%), whereas the smallest participation was observed in coniferous forests (11-23%). this probably results from greater diversification of the tree-stand and the richness of available substrate. macromycetes in selected forest communities 93 references abromeit j. 1905. bericht über die 43 jahresversammlung in culm am 7. october 1904. schr. physik.ökon. ges. königsberg 46: 50–83. bajkiewicz-grabowska e. 1989. charakterystyka geograficzna i klimatyczna mazurskiego parku krajobrazowego oraz podział hydrograficzny na wydzielone zlewnie jeziorne. 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(in:) j. b. faliński, w. mułenko (eds). cryptogamous plants in the forest communities of białowieża national park. general problems and taxonomic groups analysis. (project crypto 2). phytocoenosis 7 (n.s.), archiv. geobot. 4: 125–128, 133–136, 149–151, 155–157. skirgiełło a. 1998. macromycetes of oak-hornbeam forests in the białowieża national park – monitoring studies. acta mycol. 33 (2): 171–189. wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków, 812 pp. wojewoda w., heinrich z., komorowska h. 1999. macromycetes of oak-lime-hornbeam woods in the niepołomnice forest near kraków (s poland) – monitoring studies. acta mycol. 34 (2): 201–266. macromycetes in selected forest communities 95 udział macromycetes w wybranych zbiorowiskach leśnych mazurskiego parku krajobrazowergo streszczenie w latach 1997–2000 na terenie mazurskiego parku krajobrazowego prowadzono badania mikosocjologiczne, dotyczące występowania grzybów wielkoowocnikowych (macromycetes). w niniejszej pracy przedstawiono charakterystykę mikosocjologiczną 5, wybranych zespołów leśnych: peucedano-pinetum, vaccinio uliginosi-pinetum, serratulo-pinetum, tilio-carpinetum oraz fraxino-alnetum. zastosowano ogólnie przyjętą metodę prowadzania badań mikosocjologicznych. wielkość pojedynczej powierzchni wynosiła 400 m2. w prezentowanych fitocenozach stwierdzono występowanie 335 taksonów macromycetes. na terenie parku, w trakcie badań własnych, odnotowano łącznie 354 taksony macromycetes, co z danymi z literatury daje łącznie 506 taksonów, których występowanie stwierdzono na obszarze mazurskiego parku krajobrazowego. w trakcie obserwacji mikologicznych określono skład gatunkowy badanych zespołów leśnych parku. najbogatsze w gatunki macromycetes okazały się fitocenozy tilio-carpinetum (198 taksonów) oraz serratulo-pinetum (171 taksony). potwierdza to związek liczby występujących gatunków grzybów wielkoowocnikowych z bogatym drzewostanem – różnorodność dostępnego substratu dla rozwoju grzybni. prześledzono udział grzybów z poszczególnych grup bioekologicznych w badanych zbiorowiskach leśnych. gatunki mikoryzowe stanowią od 43 do 66 % mikobioty badanych zbiorowisk borowych. w zespołach lasowych udział ich sięga niespełna 27% mikobioty badanych fitocenoz. odwrotna tendencja zaznaczyła się w przypadku saprotrofów nadrewnowych. największy udział grupa ta miała w grądach (ok. 35%) i łęgach (45%), najmniejszy w borach (11-23%). potwierdza to związek liczby gatunków nadrewnowych z bogactwem dostępnego substratu (zróżnicowany drzewostan). 2014-01-01t11:48:52+0100 polish botanical society the occurrence of amanita strobiliformis (paulet ex vittad.) bertill. in szczecin and its distribution in poland stefan friedrich department of botany and plant protection, west pomeranian university of technology in szczecin ul. słowackiego 17, pl-71-434 szczecin, stefan.friedrich@zut.edu.pl friedrich s.: the occurrence of amanita strobiliformis (paulet ex vittad.) bertill. in szczecin and its distribution in poland. acta mycol. 48 (1): 123–131, 2013 results of long-term research into the occurrence of amanita strobiliformis in szczecin are presented. five new localities of the species are described and its distribution in poland is discussed. key words: amanitaceae, urban mycobiota, fungal phenology, chorology introduction amanita strobiliformis is a thermophilous and calciphilous species that forms symbiosis primarily with deciduous trees, mostly of the genera fagus, quercus, betula and tilia, less frequently with coniferous trees: pinus and picea. it is widespread and occurs in australia, east and central asia, north america, north africa and europe. in europe, it occurs more frequently in the south and its northern limit reaches southern scandinavia but does not exceed 60° latitude (krieglsteiner 2003). amanita strobiliformis was thought to be extinct in poland (wojewoda, ławrynowicz 1992) as it was initially known only from tomaszów lubelski (błoński 1896) and from elbląg (nitardy 1904). the herbarium specimen from the elbląg locality is not available and skirgiełło (1972) suggested it might have been a determination error. the species has been recorded by several authors at scattered localities throughout poland since the mid 1990s and is included as a rare species (r) on the current red list of fungi (wojewoda, ławrynowicz 2006). the occurrence of amanita strobiliformis in szczecin between 1997 and 2012 is characterized below. five new localities recorded in the years 2007-2011 are presented and the current distribution of the species in poland is discussed. acta mycologica vol. 48 (1): 123–131 2013 doi: 10.5586/am.2013.014 124 s. friedrich materials and methods amanita strobiliformis was reported from the dendrological garden in szczecin where it was first recorded in 1997 (friedrich, orzechowska 2002; friedrich 2011). it was the sixth locality of the species in poland. both the locality and the dendrological garden was systematically observed after the first record for the occurrence of mycobiota. fruit bodies of amanita strobiliformis were counted every year and the geographical position of the occurrence sites was determined in the field and mapped. a spread of amanita strobiliformis from the initial site and an increased range were recorded. fruit bodies occurred only at one site covering an area of ca. 20 m2 in the first four years (site 1). they were also recorded outside site 1 in the following years and occurred at a total of twelve field sites throughout the study. they grew at nine sites in the dendrological garden in szczecin and its immediate vicinity in the final year (2012). at present the occurrence area of amanita strobiliformis resembles a triangle 560 × 580 × 350 m and is regarded as one locality comprising twelve sites in the discussion. the farthest points from site 1 are 180 m westwards, 410 m north-eastwards and 350 m eastwards. the distance between adjacent sites of this vast locality is between 20 and 50 m. the locality in the dendrological garden in szczecin the locality: characteristic features. site 1 of the locality where amanita strobiliformis has fruited every year since 1997 is in the south-east corner of the dendrological garden and is surrounded by a small stone wall on three sides. the wall borders on an asphalt park alleyway, a street słowacki and the premises of the west pomeranian university of technology (formerly agricultural academy). an old tree stand which remains after a former cemetery stand consists of quercus robur, tilia cordata, acer pseudoplatanus, platanus × hispidus and pseudotsuga menziesii. fruit bodies of a. strobiliformis occurred between two oaks and a lime tree. the stand belongs to forest communities of the class querco-fagetea but only aegopodium podagraria, ficaria verna and the moss plagiomnium undulatum were recorded. the forest floor is not dense. as well as the above species, it mostly comprises species of synanthropic communities of the class artemisietea (alliaria petiolata, anthriscus sylvestris, chaerophyllum temulum, galium aparine, geranium robertianum, geum urbanum, viola odorata) and stelarietea (lapsana communis, veronica hederifolia) and species of meadow communities of the class molinio-arrhenatheretea (achillea millefolium, arrhenatherum elatius, bellis perennis, ceratium holosteoides, dactylis glomerata, festuca pratensis, poa pratensis, taraxacum officinale, trifolium pratense, t. repens). these species are accompanied by allium vineale, hedera helix, moechringia trinervia, plantago major, poa annua and veronica chamaedrys. the forest floor is mown, usually twice every year. this sometimes contributes to the destruction of fungal fruit bodies. the litter of varying thickness and single specimens of few species of herbaceous plants and moss species occur at other sites outside site 1 on the lawn in the west pomeranian university of technology. the occurrence of amanita strobiliformis in poland 127 more fruit bodies matured in favourable weather conditions, especially 2010-2011, and the period of their full development, from the development of the primordium to the initial phase of drying or dying, lasted from six to nine days, usually seven days. many fruit bodies dried at different stages of development and did not produce spores in hot years with a rain-free summer period, especially in 2006. the period of the full development observed in few fruit bodies lasted then even 12 days. first fruit bodies fig. 4. localities of amanita strobiliformis in the atpol grid square system: 1 – ac-41 – słowiński national park, czołpino (bujakiewicz, lisiewska 1983), 2 – ac-97 – ostrzyce (new locality), 3 – ac-89 – gdańsk (wilga 2004, 2005), 4 – ad-80 – gdańsk, ul. narutowicza (wilga 2004, 2005), 5 – ad80 – gdańsk, abrahama and siemiradzkiego (kujawa, gierczyk 2010), 6 – bd-05 – elbląg (nitardy 1904), 7 – bf-00 – węgorzewo (new locality), 8 – ba-73 – szczecin, dendrological garden (friedrich, orzechowska 2002), 9 – ca-16 – przelewice (new locality), 10 – ca-30 – bielinek (new locality), 11 – db-08 – poznań, botanical garden, adam mickiewicz university (lisiewska, mikołajczak 1998), 12 – db-08 – poznań, sołacki park (kudławiec 2008, 2009, 2010, 2011), 13 – db-48 – dezydery chłapowski landscape park (kujawa 2009), 14 – eb-61 – góry kaczawskie mts, „góra miłek” reserve (narkiewicz 2000), 15 – ee-73 – góry świętokrzyskie mts, góra malik (łuszczyński 2002, 2007, 2008), 16 – fb-36 – kotlina kłodzka basin, góra bielica (kujawa 2005), 17 – fb-36 – kotlina kłodzka basin, góra słupiec (kujawa 2005), 18 – fb-47 – kotlina kłodzka basin, bolesławów (kujawa 2005), 19 – fd-17 – giebło (new locality), 20 – fg-04 – tomaszów lubelski (błoński 1896), 21 – gf-03 – miejsce piastowe (hreczka 2010), 22 – gf-21 – olchowiec (kujawa, gierczyk 2010). 128 s. friedrich were observed on the 13th june (2007) at the earliest and the last ones on the 7th october (2008) at the latest during the study period. the fruiting peak occurred in july when 53% of fruit bodies were recorded and in august – 30% (figs 2, 3). the longest fruiting period lasting three months was observed in 2011 (from the 5th july until the 5th october) and in 2012 (from 18th june until 18th september). some fruit bodies grew in pairs. this was observed for 56 fruit bodies, i.e. ca. 16% of their total number. amanita strobiliformis is endangered by slugs feeding on its fruit bodies. a threat is also posed by people who thoughtlessly destroy many fungal species. a total of 108 fruit bodies (ca. 31%) were destroyed during the study period: 56 were eaten by slugs and 52 were destroyed by people. slugs posed a smaller risk in dry years when heavy blackening of fruit bodies by insect larvae was observed more frequently. new localities. five new localities of amanita strobiliformis scattered across poland were recorded between 2007 and 2011. three are in various beech communities while the other two are in a city park and a dendrological garden. the localities are characterized below in the order they were reported. they are positioned in relation to the nearest village or town in the atpol grid (zając 1978) modified for mycology by wojewoda (2000), in geographic mesoregions (kondracki 2002) and in voivodeships. the numbering of localities corresponds to the numbering in figure 4 which also presents localities known from the literature. przelewice (fig. 4 – locality 9). atpol ca-16, równina pyrzycko-stargardzka plain (313.31), west pomeranian voivodeship. the locality of amanita strobiliformis in przelewice was recorded during long-term systematic research into the mycobiota of the dendrological garden. a total of 14 fruit bodies were observed over three years (18.06.2007, 19.07.2007, 08.07.2009, 10.08.2009, 12.07.2010, 20.07.2011). they grew at four different sites less than a hundred meters apart. the majority of fruit bodies grew near fagus sylvatica and fraxinus exscelsior, and two near betula lutea, prunus serotina and malus sp. six fruit bodies were destroyed by people despite the monitoring in the garden. bielinek (fig. 4 locality 10). atpol ca-30, pojezierze myśliborskie lake district (314.41), west pomeranian voivodeship. amanita strobiliformis was observed only once (20.09.2008) at the locality in the bielinek xerothermic reserve, ca 2 km ne of bielinek. three fruit bodies grew under fagus sylvatica by a path along the margin of galio odorati-fagetum which covers a gentle south-facing slope. węgorzewo (fig. 4 – locality 7). atpol fb-00, kraina wielkich jezior mazurskich (great masurian lakes region, 842.83), warmia and mazury voivodeship. this locality of amanita strobiliformis was recorded during a holiday stay in węgorzewo on 27.07.2009. it is in the helwing park, near generała j. bema street and a few meters from a park alleyway. three developing fruit bodies grew on a lawn near tilia cordata, just outside the reach of the crown. giebło (fig. 4 – locality 19). atpol df-17, wyżyna częstochowska upland (341.31), silesian voivodeship. the locality of amanita strobiliformis was recorded in the las niwiski forest in giebło village near ogrodzieniec, ca. 6 km nw of the pilica river. fagus sylvatica-cruciata glabra forest community occurs at the locality. one fruit body was recorded on the soil under fagus sylvatica in september 2008 and 2009 (leg. et det. d. karasiński). one fruit body was destroyed by mushroom pickers. the occurrence of amanita strobiliformis in poland 129 ostrzyce (fig. 4 – locality 2). atpol ac-97, pojezierze kaszubskie lake district (314.51), pomeranian voivodeship. amanita strobiliformis was observed in the ostrzycki las reserve, ca. 10 km sw of kartuzy, in fagus sylvatica-cypripedium calceolus, on brown rendzina developed on a layer of calcareous mud. five fruit bodies at different stages of development grew on the soil under fagus sylvatica on 14.08.2010 (leg. et det. d. karasiński). discussion amanita strobiliformis was recorded in poland only at five localities in the 20th century. it was noted for the first time after a period spanning a few decades in 1967 (bujakiewicz, lisiewska 1983). it is not clear whether the species did not occur (did not develop fruit bodies) in poland or whether it was not observed for various reasons for such a long time in the 20th century. some 90 localities were recorded in germany (krieglsteiner 1991) in similar physiogeographic conditions (between 50° and 55° latitude) in the same period. one of the reasons for this may be that mycological research in poland was mostly conducted in natural forests, national and landscape parks, nature reserves while this species often occurs in parks and anthropogenic areas in poland. descriptions of localities of amanita strobiliformis in poland available in the literature show that half of the localities are in park-like anthropogenic tree patches, e.g. the botanical garden in poznań, dendrological gardens in szczecin and przelewice, city parks in poznań, węgorzewo and gdańsk. fruit bodies usually occur under fagus sylvatica, less frequently under quercus sp., tilia sp. and betula pendula. as fruit bodies develop in parks, in noticeable places, they are destroyed by people, as noted by authors such as wilga (2005) and kudławiec (2008-2011). at present a total of 22 localities, including the new localities described here, in 19 atpol 10x10 km squares are known in poland (fig. 4). approximately half of them have been reported by non-professionals in the last ten years. amateur fungal groups importantly contribute to the recognition of the distribution of fungi in poland, also rare and threatened species. fruit bodies were produced between june and october at the localities in poland. this was also the period when fruit bodies were observed at the locality in the dendrological garden in szczecin over a period of 16 years. a detailed seasonal rhythm of fruit body production at this locality is similar to the results of studies in germany (baden-württembergs) but the fruiting peak begins a month earlier (krieglsteiner 2003). one-time data on the occurrence of amanita strobiliformis which usually produced only single fruit bodies (1-3) are reported from the majority of polish localities. the most abundant localities where amanita strobiliformis produces annually from over ten to over fifty fruit bodies are the dendrological garden in szczecin and the park sołacki park in poznań, where a total of over 50 fruit bodies were observed over four consecutive years (kudławiec 2008, 2009, 2010, 2011). acknowledgements. the author thanks dr. dariusz karasiński for providing information on two new localities of amanita strobiliformis. 130 s. friedrich references błoński f. 1896. przyczynek do flory grzybów polski. pamiętn. fizyjogr. 4 (iii): 63-93. bujakiewicz a., lisiewska m. 1983. mikoflora zbiorowisk roślinnych słowińskiego parku narodowego. bad. fizjogr. pol. zach. seria b – bot. 34: 49-77. friedrich s. 2011. charakterystyka występowania muchomora szyszkowatego amanita strobiliformis (paulet ex vittad.) bertillon w szczecinie. konferencja naukowa. polskie tradycje użytkowania grzybów oraz ich ochrony wkładem do europejskiego dziedzictwa kultury. łódź 3-5.11.2011. wyd. uniw. łódzkiego, łódź: 45-46. friedrich s., orzechowska m. 2002. macromycetes w środowisku miejskim szczecina. bad. fizjogr. pol. zach. seria b, bot. 51: 7-30. hreczka a. 2010. amanita strobiliformis. id 169498. w: snowarski m. atlas grzybów polski. rejestr gatunków grzybów chronionych i zagrożonych. http://www.grzyby.pl./rejestr-grzybow-chronionychi-zagrozonych.htm kondracki j. 2002. geografia regionalna polski. pwn, warszawa. krieglsteiner g.j. 1991. verbreitungsatlas der grosspilze deutschlands (west). verlag eugen ulmer. krieglsteiner g.j. (hrsg.). 2003. die grosspilze baden-württembergs. b. 4. ständerpilze: blätterpilze ii. ver. eugen ulmer, stuttgart. kudławiec b. 2008. amanita strobiliformis. id 114510. w: snowarski m. atlas grzybów polski. rejestr gatunków grzybów chronionych i zagrożonych. http://www.grzyby.pl./rejestr-grzybow-chronionychi-zagrozonych.htm kudławiec b. 2009. amanita strobiliformis. id 156115. w: snowarski m. atlas grzybów polski. rejestr gatunków grzybów chronionych i zagrożonych. http://www.grzyby.pl./rejestr-grzybow-chronionychi-zagrozonych.htm kudławiec b. 2010. amanita strobiliformis. id 195538. w: snowarski m. atlas grzybów polski. rejestr gatunków grzybów chronionych i zagrożonych. http://www.grzyby.pl./rejestr-grzybow-chronionychi-zagrozonych.htm kudławiec b. 2011. amanita strobiliformis. id 195538. w: snowarski m. atlas grzybów polski. rejestr gatunków grzybów chronionych i zagrożonych. http://www.grzyby.pl./rejestr-grzybow-chronionychi-zagrozonych.htm kujawa a. 2005. „rejestr gatunków grzybów chronionych i zagrożonych” – nowa forma gromadzenia danych mikologicznych pochodzących od amatorów. podsumowanie roku 2005. przegląd przyrodniczy 16 (3-4): 17-52 kujawa a. 2009. macrofungi of wooded patches in the agricultural landscape. i. species diversity. acta mycol. 44 (1): 49-75. kujawa a., gierczyk r. 2010. rejestr gatunków grzybów chronionych i zagrożonych w polsce. iii. wykaz gatunków przyjęty do rejestru w roku 2007. przegląd przyrodniczy 21 (1): 8-53. lisiewska m., mikołajczak m. 1998. ogród botaniczny uniwersytetu im. a. mickiewicza w poznaniu jako środowisko przyrodnicze grzybów wielkoowcnikowych. bad. fizjogr. pol. zach. seria b, bot. 47: 7-44. łuszczyński j. 2002. preliminary red list of basidiomycetes in the góry świętokrzyskie mts (poland). polish bot. j. 47 (2): 183-193. łuszczyński j. 2007. diversity of basidiomycetes in various ecosystems of the góry świetokrzyskie mts. monogr. bot. 97: 1-218. łuszczyński j. 2008. basidiomycetes of the góry świetokrzyskie mts a checklist. wyd. uh-p, kielce. narkiewicz c. 2000. borowik szatański boletus satanas i muchomor szyszkowaty amanita strobiliformis w rezerwacie „góra miłek” w górach kaczawskich. przyr. sudetów zach. 3: 69-72. niedźwiecki e., meller e., malinowski r., sammel a., kruczyńska j. 2004. mniszek pospolity (taraxacum officinale) jako bioindykator zanieczyszczenia metalami ciężkimi gleb miejskich szczecina. folia univ. agric. stetin. agricultura 242 (98): 103-108. nitardy e. 1904. die kryptogamenflora des kreises elbing, hedwigia 43: 314-342. skirgiełło a. 1972. materiały do poznania rozmieszczenia geograficznego grzybów wyższych w europie. iv. acta mycol. 8 (2): 191-218. wilga m. 2004. chronione i zagrożone grzyby wielkoowocnikowe (macromycetes) trójmiejskiego parku krajobrazowego (pomorze gdańskie). przegląd przyrodniczy 15 (1-2): 3-17. the occurrence of amanita strobiliformis in poland 131 wilga m.s. 2005. muchomor szyszkowaty amanita strobiliformis (paulet ex vittad.) bertillon w gdańsku. chrońmy przyrodę ojcz. 61 (5): 93-100. wojewoda w. 2000. (ed.) atlas of the geographical of fungi in poland. 1. w. szafer institute of botany, polish academy of sciences, kraków. wojewoda w., ławrynowicz m. 1992. red list of threatened macrofungi in poland. (in:) k. zarzycki, w. wojewoda, z. heinrich (eds). list of threatened plants in poland, 2 ed. w. szafer institute of botany, polish academy of sciences, kraków: 27-56. wojewoda w., ławrynowicz m. 2006. red list of the macrofungi in poland. (in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. 3 ed. w. szafer institute of botany, polish academy of sciences, kraków: 53-70. zając a. 1978. założenia metodyczne „atlasu rozmieszczenia roślin naczyniowych w polsce”. wiad. bot. 22 (3): 145-155. występowanie muchomora szyszkowatego amanita strobiliformis (paulet ex vittad.) bertill. w szczecinie na tle jego rozprzestrzenienia w polsce streszczenie w pracy przedstawiono wyniki 16-letnich (1997-2012) badań nad występowaniem muchomora szyszkowatego amanita strobiliformis w ogrodzie dendrologicznym w szczecinie. w 12 punktach tego rozległego stanowiska wyrosło łącznie 347 owocników, z czego aż około 36% w punkcie 1, w którym owocniki obserwowano rokrocznie (fig. 1). w pozostałych punktach owocniki wyrastały najczęściej przez 3-4 lata (od 1 roku do 7 lat). najwcześniej owocniki obserwowano 13 czerwca, a najpóźniej – 7 października. szczyt owocnikowania przypadał w lipcu, kiedy stwierdzono 53% owocników i sierpniu – 30% (figs 2, 3). najdłuższy okres owocnikowania trwający 3 miesiące, wystąpił w latach 2011 i 2012. przedstawiono również 5 nowych stanowisk muchomora szyszkowatego stwierdzonych w różnych regionach kraju, w latach 2007-2011. łącznie z tymi nowymi stanowiskami, w polsce znane są obecnie 22 stanowiska zlokalizowane w 19 kwadratach atpol 10 × 10 km (ryc. 4). 2013-06-22t22:33:05+0100 polish botanical society 2014-08-25t14:56:47+0200 polish botanical society influence of some physical factors on the growth and sporulation of entomopathogenic fungi jerzy piątkowski and aneta krzyżewska institute of genetics and microbiology, university of wrocław przybyszewskiego 63/77, pl-51-148 wrocław, jurekp@microb.uni.wroc.pl p i ą t k o w s k i j., k r z y ż e w s k a a.: influence of some physical factors on the growth and sporulation of entomopathogenic fungi. acta mycol. 42 (2):255-265, 2007. the objective of the study was to examine, how some physical factors and inoculation techniques affect the growth and sporulation of three insecticidal fungal strains, hirsutella tompsonii, paecilomyces sp., and pandora sp. although the methods of inoculation, as well as such physical parameters as temperature, the osmotic value of the habitat and uv radiation, exerted an influence on the above-mentioned features of the fungi, their contribution varied from one strain to another. some of the effects, however (e.g., the production of filamentous structures by pandora, or the lethal action of glycerol on pandora), require a close examination. key words: entomopathogenic fungi, phisical factors, sporulation, growth of fungi introduction apart from unquestionable advantages, the application of insecticidal chemicals has a number of major drawbacks. these substances are toxic not only to the target insects but to the beneficial ones as well. they are also known for their harmful effect on human health. during long-term exposure, the target insects develop resistant forms to the insecticide applied. in the process of biotransformation by bacteria, a variety of new compounds may be produced, including cancerogens (r ó ż a ń s k i 1992). for these reasons, many investigators have directed their attention to the use of a natural enemy of a pest as a less harmful environmental pollutant. of those investigations, the one into the entomopathogenic fungi as manipulative tools in biological control deserves special consideration. in general, the fungi are able to grow under laboratory conditions (p a t e r s o n et al. 1987) but the production of efficient and inexpensive commercial preparations require mastering the competence for an effective and low-cost cultivation of the fungi, as well as for making them produce a great amount of spores. the efforts of the scientists have also focused on the secondary acta mycologica vol. 42 (2): 255-265 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 256 j. piątkowski and a. krzyżewska metabolites released by the fungi, which certainly play an important role in the process of infection and killing pests. all that mean, that the production of efficient preparations on the entomopathogenic fungi basis requires information about the biology of the organisms, their nutritional needs and their response to physical factors. spore formation, germination, hyphal and spore survival are influenced by temperature, humidity, light, organic pollutants, salinity, antagonists, etc. (c a r r u t h e r s , h u r a l 1990). investigations performed so far have revealed that the response of the fungi to the habitat factors depends on the species and ecotype involved and that the activity of the fungi against their host follows a seasonal pattern. thus, beauveria bassiana infects leptinotarsa decemlineata more often in the summer and paecilomyces farinosus in the autumn and winter seasons (b a j a n , k m i t o v a 1997). with entomophthora destruens and conidiobolus thromboides, the extent of spore production depends on the season of the year, or (to be more exact) probably on the length of daylight (k r e j z o v a 1988). an important factor, both inhibiting and stimulating the growth of the organisms, is temperature, which seems to affect directly the ability to infect (m i ę t k i e w s k i et al. 1994). it is a well established fact that the optimal temperature for the species of the genus entomophthora (e.g., e. ignobilis, e. obscura, e. exitialis, or e. virulenta), which provides their most dynamic growth, ranges between 24 and 27°c (h a l l , b e l l 1961). a similarly stimulating effect of this temperature range on the growth of mycelium has been reported for the genera beauveria, metarrizium, and paecilomyces (h a l s w o r t h , m a g a n 1999). further study has revealed that the optimal temperatures for hyphal survival are identical with those for the survival of spores (b r o b y n et al. 1985). the temperature factor affects the duration of sporulation; e.g., in erynia neoaphidis the temperature of 5°c inhibits sporulation completely (d r o m p f et al.1998). the spores of entomophthora apiculata, e. virulenta and e. coronata are deactivated by extreme temperatures –8°c and +36°c (ye n d o l 1968). it has been reported that hit shock proteins appear in entomopathogenic fungi and that their appearance is induced by the habitat of 45°c (x a v i e r , k h a c h a t o u r i a n s , 1996). humidity exerts a direct influence on fungi, both on vegetative cells and spores. the optimal relative humidity oscillates within a wide range, depending on the species involved. for example, entomophthora aphidis and e. thaxteriana discharge spores only when humidity ranges between 70 and 90%. under such conditions, the spores survive a few days, but in cooler air – several weeks or even months (w i l d i n g 1973). in the majority of the genera, germination is inhibited even at 90% or lower humidity, but in some entomophthora conidium may germinate and infect insects at humidity lower than 50% (u z i e l , k e n n e t 1991). in many fungi, the 24-hour rhythm of spore production is daylight-dependent. light accounts for a variety of effects. it has been found that the extent of sporulation in entomophthora virulenta is significantly higher in darkness than in light under laboratory conditions (l a t g e et al. 1978), in contrast to entomophthora spheroderma which discharges more conidium during daylight, both in vivo and in vitro. light-dependent periodicity of spore release has also been reported for erynia radi influence of physical factors 257 cans. this periodicity appeared after exposure of a culture alternately to light and dark (ya m a m o t o , a o k i 1983). there are not very many references to the influence of physical factors on entomopathogenic fungi in the available literature, although fungi are known to be strongly affected by this kind of factors. so far, the reports on these fungi have focused on the optimal growth medium for cultivation and on the basic biological properties, especially those involved in such physiological activities as sporulation, production of secondary metabolites and infection of the host insect. the objective of the experimental study reported on in the present paper was to throw new light on the variety of physical and environmental factors which affect the entomopathogenic strains belonging to different genera. for the purpose of our study, the genera paecilomyces, pandora and hirsutella were chosen. the fungi under study differ in the morphology of their mycelium, in resistance to some unfavourable external factors and in the mode of sporulation. the chosen fungi represent different unrelated genera, thus enabling the verification of whether the observed effects produced by the physical factors being tested are common to all fungi or depend on the composition and physiological properties of a particular fungal organism. materials and methods strains used in the investigations: pandora sp. isolated from the aphids protrama ranunculi; paecilomyces sp. from a larva of the family cantharidae, and hirsutella thompsonii fischer from phylocoptura oleivora. all the strains were obtained by courtesy of prof. bałazy, research centre for agricultural and forest environment, polish academy of sciences, poznan. measurement of the rate of mycelium growth: round discs of mycelium (4 mm in diameter) were cut out from the 7-day preculture on the solid ypg medium (yeast extract 1%, bacto pepton 1%, glucose 2%, bacto agar 2%). each disc was placed at the central point of the medium in a petri dish and incubated at 22°c. mycelium growth was measured every 24 hours. relation between mycelium growth and inoculation method. the mycelium discs were translocated into a tube with 0.5 ml saline (0.9%nacl), and afterwards ground against the wall of the tube to obtain sols of fragmented mycelium. the solution prepared in this way was dropped onto the central part of the solid ypg medium, spread over the whole medium surface, or streaked with a reductive streack using a loop. the mycelium discs were also placed on the solid medium without previous grinding. in order to observe the growth of the mycelium inside the solid medium, the discs were flooded (after being placed on the solid medium) with the same, still warm enough (50°c) liquid medium. effect of low temperature: the experiments were performed with the following media: saline (0.9 %nacl) – medium a; fluid medium for the storage of bacterial strains consisting of 1% trypton, 1% yeast extract and 0.5% nacl, 1000 ml of this medium were treated with 450 ml of 50% glycerol – medium b; medium for the storage of yeast cells consisting of yeast extract,10g, bacto pepton, 10g, glucose, 20g, glycerol, 250 ml and distilled water, 750 ml – medium c. 0.5 ml of the liquid media was poured into the eppendorf tubes and thereafter 4 mm discs of fungal mycelium influence of physical factors 259 the method of hyphal inoculation into the growth medium was also an important factor to which each of the three strains responded differently. pandora was growing in the form of a colony, when the suspension of the previously ground mycelium was dropped at the centre of the solid medium or spread over the whole surface of the medium by means of a spreader. pandora placed on the plate surface with a reductive streak using a loop showed a poorer growth when the cells were taken from the surface of the fungus or from the suspension of ground mycelium in saline. there was no growth at all when the pandora cells were flooded with the medium, which implies that with pandora deep inoculation is without any meaning. on the other hand, hirsutella and paecilomyces were growing quite well regardless of the inoculation method applied. only the paecilomyces sp. strain changed the morphology of its colony when plated deeply. the colony was plain but the pink colour typically occuring on the mycelium was no longer produced. in order to verify the viability of the fungi at low temperature, a saline (0.9% nacl) and a glycerol medium was used for the storage of the microorganisms at +6, -18, -72°c, the temperature for the control being 22°c (tabs 1, 2 and 3). as shown by these data, pandora sp. exhibited a very high sensitivity to the negative temperatures used. this strain was able to survive 7 weeks when kept in saline ta b l e 1 growth and sporulation of pandora sp. related to time of exposure to various temperatures for different media time 22°c 6°c -18°c -72°c medium 24 hours ++ +++ spores ─ + a 7 days ++ +++ ─ + a 7 weeks ─ ─ ─ ─ a 24 hours +++ spores +++ spores + ─ b 7 days +++ spores +++ spores ─ ─ b 7 weeks ─ ─ ─ ─ b 24 hours + +++ ─ ─ c 7 days ─ ─ ─ ─ c 7 weeks ─ ─ ─ ─ c ta b l e 2 growth and sporulation of paecilomyces sp. related to time of exposure to various temperatures for different media time 22°c 6°c -18°c -72°c medium 24 hours ++ +++coremia +++ +++ coremia a 7 days ++ ++ ++ +++ coremia a 7 weeks + +++ ++ coremia +++ coremia a 24 hours +++ +++ +++ +++ coremia b 7 days +++ + +++ coremia +++ coremia b 7 weeks ++ + ++ +++ b 24 hours +++ +++ +++ coremia +++ c 7 days ─ +++ +++ +++ c 7 weeks + ++ ++ +++ 260 j. piątkowski and a. krzyżewska at room temperature, and only 7 days when stored in the glycerol medium. after 24 hours of incubation in saline at 6°c, there was an induction of spore formation, but no sporulation occured when the time of incubation was extended to 7 days. it is interesting to note that in the glycerol medium sporulation becomes visible after 24 hours and 7 days of incubation. the paecilomyces sp. strain was found to be more resistant to the temperature used in the experiments, it survived within the temperature range of 6°c to minus 72°c. a longer exposure to the deficiency of nutrients in the habitat had an adverse effect on the ability of the strain to reconstruct a colony. furthermore, as it can be infered from table 2, at some of the temperatures and incubation times applied, the induction of an asexual reproduction structure was observed in paecilomyces. these structures (referred to as coremia) appeared after different time at minus 18°c and depended on the type of the medium used. coremia also appeared at each incubation time in saline at minus 72°c, as well as after 24 hours and 7 days of incubation in the glycerol medium at minus 72°c. these structures did not form when the strain was kept at minus 72°c in the glycerol medium for yeast storage and when the temperature of incubation was 22°c, as well as in any medium variant at 6°c. as for hirsutella thompsonii, full growth occured after keeping this fungus in whatever variant of temperature, medium or time except 7 weeks in the glycerol medium for yeast. in this medium, hirsutella survived 7 weeks, but only if temperature was drastically decreased. this finding is important to the storage of these strains. the glycerol medium for the storage of bacterial and yeast strains is used with success at temperatures even as low as minus 70°c. as expected, paecilomyces and hirsutella were able to endure such temperature even for 7 weeks. but as it can be infered from table 1, pandora did not survive under these conditions, even though the habitat was less drastic: 24 hours of incubation and minus 18°c. in order to explain the unexpected behaviour of pandora, we decided to examine the effect of different glycerol concentrations on the rate of survival. to eliminate the influence of the water crystals that form at low temperature and may account for the killing of the cells, use was made of room temperature (22°c) alone. the glycerol concentrations of choice were 30; 15; 7.5; and 3.25%. after incubation of the pandora mycelium in these variants of glycerol solutions, the discs were ta b l e 3 growth and sporulation of hirsutella tompsonii related to time of exposure to various temperatures for different media time 22°c 6°c -18°c -72°c medium 24 hours +++ +++ +++ +++ a 7 days +++ +++ +++ +++ a 7 weeks ++ ++ ++ ++ a 24 hours +++ +++ +++ +++ b 7 days +++ +++ +++ +++ b 7 weeks ++ ++ ++ ++ b 24 hours +++ +++ +++ +++ c 7 days +++ +++ +++ +++ c 7 weeks ─ ─ + ++ c 262 j. piątkowski and a. krzyżewska to fragment. the clod was surrounded by a delicate mycelium, which was easy to fragment. the adjacent zone, again, had the shape of a coiled string. the whole structure displayed a characteristic colony pattern. when the mycelia were treated with glycerol, this was parallelled by a delayed production of spores which did not appear until the ninth day after the culture on the solid full medium had been started. the inhibition effect exerted by these glycerol concentrations implies that the fungus pandora is exceptionally sensitive to water tranfer from the cells. so, there were performed tests with different concentrations of nacl, including those producing a hypertonic environment, which remove part of the water from the cells. the results of the tests are plotted in figure 3. in general, the rate of growth decreases with the increasing osmotic value of the medium in which the fungi were kept before being translocated onto the ypg medium. but surprisingly, as the time of incubation in nacl increases, so does the vitality of the mycelium translocated from nacl onto the solid full medium. after 24 hours of incubation in the hypotonic solutions (0.006m and 0.06m), the strain was growing as fast as in the control. the results of irradiating the fungal mycelium with uv rays indicate that the uv rays did not affect the dynamics of growth in any of the strains. only a change in the colour of the paecilomyces and hirsutella colony surface was noticed. with paecilomyces irradiated for 60 min., the typical pigment did not appear after 9 days of irradiation. interestingly, on the days that followed the pigment alternately appeared and disappeared. such phenomenon was not observed in any culture of the other investigated species. discussion entomopathogenic fungi form a highly inhomogeneous group of eucaryotic organisms. the comparison of the three species (differing morphologically and unrelated in the sense of not belonging to the same rod) revealed strong differences in the rate of growth. pandora grew much faster than the other species. but all of the strains grew fast enough to enable a comparative study of how the various factors affected the growth process. after 20-day observation of the increase in the diameter of hirsutella, when the mycelium attained a diameter five times as large as the initial one, it was possible to examine the influence of the environment on microorganism growth, colony morphology, sporulation process and secondary metabolites production. our study also substantiated the importance of the method by which the samples of the cells were inoculated on/in the medium. tearing off fragments of the mycelium and placing them on/in other media was very useful when starting a new culture. in some instances it is necessary to develop growth on the whole surface of a solid medium over a short period, and this is easy to achieve if we have a solution of spores. we started a new culture in order to prepare a solution of small fragmented mycelium and inoculate it afterwards. all the three fungi were found to grow fast after such inoculation. pandora showed greater dynamics of sporulation under such conditions, probably owing to the mechanical stress triggered by the tearing-off of the micelium, which induced the sporulation process. influence of physical factors 263 the fungi under study responded differently to the method of placing them on the medium. deep inoculation obtained by pouring the mycelium over a melted agar medium inhibited the growth of pandora, while hirsutella displayed no significant changes either in the morphology or the size of the colony. morphological changes were detected in paecilimyces. compared to the control its colony was plain, without the characteristic pigment. these findings have led to the conclusion that the method of inoculation plays a particular role in the investigations of fungi. an important requisite for laboratory experiments with microorganisms (also with fungi) is storing them at low temperature as museum strains. in many instances it is useful to keep them at 72°c in the glycerol medium. our results, however, show that this procedure fails to be sufficient for pandora. after 7-day storage in saline or in the glycerol medium for bacteria, the fungus survive at 6°c; it did not survive when kept in the glycerol medium for yeast over the same period. the question arises as to why pandora shows greater sensitivity when kept in the glycerol medium for yeast storage. in this medium glycerol concentration is higher than in the ypg medium and bacto pepton has been replaced with bacto trypton. this indicates that the pandora strain is very sensitive even to small changes in the composition of the medium. the paecilomyces mycelium survived in each of the media used in our study, even after 7 weeks of incubation at minus 72°c. it is interesting to note that after incubation in saline at minus 18°c we observed the appearance of the spore producing structure referred to as coremia, but this process was affected by the composition of the medium. in the medium for bacteria and in the glycerol yeast medium coremia appeared after 7 days and after 24 hours of incubation, respectively; the structures in saline did not arise after 6 weeks. these findings imply that even small changes in the composition of the medium and in the physical environment have to be considered when establishing the optimal conditions for the induction of sporulation in paecilomyces. it suffices to keep the strain at 72°c in saline, not in the glycerol medium for yeast. as for hirsutella, the strain survives when incubated in any of the medium variants used, but surprisingly not at plus 22°c in the glycerol medium for yeast over the incubation period of 7 weeks. our study of the low temperature effect suggests that glycerol might be the factor which negatively affects the viability of pandora. more precise testing methods supported this suggestion. even a 30% concentration of glycerol was able to kill the cells. glycerol concentrations below 30% decreased the rate of colony growth and induced the appearance of unique morphological mycelium structures. in order to describe the mechanism governing the impact of glycerol on pandora we should take into account the fact that one molecule of glycerol absorbes three molecules of water, and this is what explains the removal of three water molecules from the cell interior. anyway, it is difficult to accept that glycerol is transported in such amounts that enable a significant level of dehydration to be attained. of greater importance seems to be the interaction of glycerol outside the cell. in terms of molar concentrations, these were the values of 0.35m, 0.8m and 1.6m at which the rate of zoophthora growth slowed down. sodium chloride as physiological liquid is considered to be iso-osmotic to yeast cells at the concentration of 0.15 m. this means that all the inhibiting solutions of glycerol were hypertonic, and therefore capable of removing water from the pandora cells. the question is, does the fungus really have such a high sensitivity to water removal ? 264 j. piątkowski and a. krzyżewska the question has partly been answered in the course of our experiment involving various concentrations of sodium chloride. as expected, the more hypertonic was the habitat sorrunding the mycelium of pandora, the slower became the growth rate. although nacl was found to act as a growth inhibiting factor, it did not kill the fungi cells. uv radiation tests showed that exposure to uv rays did not affect the rate of growth of the mycelium. this was not surprising since the rays did not penetrate the cells in the lower parts of the colony, so they might have grown. while performing the experiments, it was interesting to observe the surface of the mycelium because the uv rays might have induced mutations changing its morphological traits. it is not unlikely that this factor could induce the production of telomorphic forms. changes in the morphology of the surface were really noticed but they all pertained solely to the colour of the mycelium. paecilomyces exposed to uv radiation for 60 min. developed the characteristic pigment only after 10 days of cultivation, but after another two days the pigment disappeared. hirsutella lost the pigment in each time variant of uv radiation. the shape of the mycelium edge became less regular. the morphology of pandora did not change as a result of exposure to uv radiation. these findings are interesting in the context of relevant literature, where the inducing effect of ionising radiation on living cells is taken into account. according to some authors (s z o t 1976), radiation energy is transferred to water molecules and afterwards translated to other compounds, producing the energization effect, which accounts for the increased activity of those compounds, including that of the enzymes. references b a j a n c., k m i t o w a k. 1997. thirty years’ studies on entomopathogenic fungi, polish ecological studies, 23: 3-4, 133–154. b r o b i n p. j., w i l d i n g n., c l a r c s. j. 1985. the persistence of infectivity of conidia of the aphid pathogen erynia neoaphidis on leaves in the field, ann. appl. biol. 107: 365–376. c a r r u t h e r s r. i., h u r a l k. 1990. fungi as naturally occurring enthomopathogens. (in:) r.r. b a k e r , p.e. d u n n (eds). new directions in biological control. alan r. liss, new york: 115–138. d r o m p h k., p e l l j. k., e i l e n b e r g j. 1998. sporulation of erynia neoaphidis from sitobion avenae cadavers, insect. pathogens and insect parasitic nematodes iobc, bulletin 21: 91–94. h a l l l. m., b e l l j. v. 1961. further studies on the effect of temperature on the growth of some entomophthorous fungi, journal of insect pathology 3: 289–296. k r e j z o v ă r. 1988. the formation and discharge of conidia in cultures of entomophthorous fungi, ćeska mycologie 42: 31–41. l a d g e l. p. 1978. growth and sporulation of entomophthora virulenta on semidefined media in liquid culture, j. invertebr. pathol. 31: 225–233. m i ę t k i e w s k i r., t k a c z u k c., z u r e k m., v a n d e r g e e s t l. p. s. 1994. temperature require-temperature requirements of four entomopathogenic fungi. acta mycol. 29 (1): 109–120. p a t e r s o n r. r., s i m m o n d s m. s., b l a n e y w. m. 1987. mycopesticidal effects of characterised extracts of penicillum isolates and purified secondary metabolites (including mycotoxins) on drosophila melanogaster and spodoptora littorali, journal of invertebrate pathology 50: 124–133. r ó ż a ń s k i l. 1992. przemiany pestycydów w organizmach żywych i środowisku. pwril, warszawa: 113–114. s z o t z. 1976. działanie promieniowania jonizującego na materię żywą. postępy techniki jądrowej, warszawa. u z i e l a., k e n n e t h r. g. 1991. survival of primary conidia and capilliconidia at different humidities in erynia (subgen. zoophthora) spp. and in neozygites freseni (zygomycotina, entomophthorales) with special emphasis on erynia radicans. j. invertebr. pathol. 58: 118–126. influence of physical factors 265 w i l d i n g n. 1973. the survival of entomophthora sp. in mummified apis at different temperatures and humidities, journal of invertebrate pathology 21: 309–311. x a v i e r i. j., k h a c h a t o u r i a n s g. b. 1996. the heat-shock response of 5 strains of entomopathogenic fungus beauveria brongniarti, canadian journal of microbiology 42: 577–585. ya m a m o t o m., a o k i j. 1983. periodicity of conidial discharge of erynia radicans. trans. mycol. soc. japan 24: 487–496. ye n d o l w. g. 1968. factors affecting germination of entomophthora conidia, journal of invertebrate pathology 10: 116–121. wpływ czynników fizycznych na wzrost i zarodnikowanie grzybów owadobójczych s t r e s z c z e n i e przetestowano wpływ wybranych czynników fizycznych oraz technik posiewu na wzrost i zarodnikowanie szczepów trzech gatunków grzybów owadobójczych: hirsutella tompsonii, paecilomyces sp. i pandora sp. zarówno technika posiewu, jak i czynniki takie jak temperatura, wartość osmotyczna środowiska, promieniowanie uv wpływają na wymienione cechy tych grzybów, chociaż wpływ ten jest różny w stosunku do poszczególnych szczepów. niektóre uzyskane efekty, np. wytwarzanie przez pandora sznurowatych struktur, czy letalny wpływ glicerolu w stosunku do pandora, powiny być poddane bardziej szczegółowym badaniom w przyszłości. 2014-01-01t11:46:39+0100 polish botanical society 2014-08-25t19:07:54+0200 polish botanical society occurrence of black truffles in poland maria ławrynowicz, tomasz krzyszczyk and marcin fałdziński department of mycology, university of łódź banacha 12/16, pl-90-237 łódź, miklaw@biol.uni.lodz.pl ławrynowicz m., krzyszczyk t., fałdziński m.: occurrence of black truffles in poland. acta mycol. 43 (2): 143–151, 2008. tuber aestivum vittad., t. mesentericum vittad. and t. bellonae quél. have been distinguished in recent collections of hypogeous fungi in poland. up till now only t. mesentericum has herbarial documentation. t. aestivum was confirmed to occur in poland. seven localities are documented with exiccates, but it is considered as a complex species. some specimens have features of t. uncinatum chat. not distinctively separated from these of t. aestivumvittad., therefore are treated as its form. the studies based on rich material support the concept of tuber bellonae as a separate species close to t. mesentericum. key words: tuber, taxonomy, ecology, distribution introduction hypogeous fungi and especially truffles have been of increasing interest in poland in the recent years. the old data on the occurrence of truffles in poland were set together by lubelska (1953). she mentions 8 species of truffles, among them tuber aestivum from 6 localities on the base of papers by: caspary (1886), błoński (1888), and alexandrowicz & błoński (1894). it was the only black truffle indicated; this taxon overlapped also t. mesentericum, but no herbarium material is now available to analyse this question. after about hundred years t. mesentericum was discovered on częstochowa upland in two localities (ławrynowicz 1988, 1990, 1999) in course of systematical searching of hypogeous fungi. these fungi produce fruit bodies only in some places called hypogeous oasas or nests, where ecological conditions are suitable. similarly, only in some years the climatic conditions unable formation of fruit bodies. the year 2007 was exceptional. the authors of this paper found independently black truffles in 13 sites. the analysis of the material permits to identify 4 taxons: tuber aestivum, t. aestivum forma uncinatum, t. mesentericum, and t. bellonae; the latter as a new species for poland. acta mycologica vol. 43 (2): 143–151 2008 144 m. ławrynowicz et al. material and methods the investigations were taken out at the calcareous hilly area, not exceeding 300 m a.s.l. on the base of type of ecosystem, density of vegetation cover, type of soil and features known as characteristic for truffle sites, the environment places for excavations were chosen. numerous collections of different hypogeous species were made during several years of field searching, but in 2007, starting from the early summer, the fruit bodies of black truffles were to be found. also in course of monitoring two above mentioned sites of tuber mesentericum two new localities of t. aestivum were discovered in july 2007. altogether more than two hundred of fruit bodies were observed by two of us (tk and mf). in some cases a dog was helping to detect the ripe carpophores. specimens were described immediately after taking out from the soil and in most cases documented at the sites concerned. fruit bodies were analysed taxonomically using the traditional procedure. the micromorphological and anatomical features of peridium cells, asci and ascospores were examined under eclypse 600 e-microscope (nikon) (200x, 400x, 1000x). materials were identified according to montecchi and sarasini (2000), riousset et al. (2001) and some other authors indicated in the “references”. the localities are indicated on three maps in the atpol system. abbreviations of collectors names: tk – tomasz krzyszczyk, mf – marcin fałdziński, mł – maria ławrynowicz. other explanations for atpol system used for hypogeous fungi are according to ławrynowicz (1989). dried specimens were deposited in the herbarium universitatis lodziensis (lod). key to species fruit body black, purpleor black-brown, with warty surface; spores reticulate 1. fruit body black becoming gray with cavity or depressions; warts small, obtuse ................................................................................................................... 2 1*. fruit body black becoming brown; warts large, acute .................................... 3 2. fruit body with distinct cavity; spores ellipsoid ..................... t. mesentericum 2*. fruit body with depressions; spores round ...................................... t. bellonae 3. gleba white becoming light yellow; spores with incomplete mesh reticulation up to 2 μm high .................................................................................. t. aestivum 3*. gleba yellowish becoming brown; spores with reticulum up to 4 μm high ....................................................................................t.aestivum forma uncinatum occurrence of black truffles 145 descriptions of the species tuber aestivum vittadini 1831 fruit bodies globose or subglobose 2-5 cm in diam.; black, becoming brown when dried. peridium strongly warty with pyramidal, irregularly polygonal warts 4-7 mm wide, truncate or depressed on top; with radial ridges and fine transversal striae. gleba fleshy, compact, at first white becoming light brown to hazel brown, at maturity marbled with white, thin veins. odour pleasant. asci 60-100 × 55-75 μm globose or pyriform, with short stalks, 2-6 spored. spores 20-45 × 15-35 μm at first hyaline becoming yellow to yellowish brown; ornamented by irregular reticulum with uncomplete alveolae, up to 2 μm high; 3-4 along the spore. remarks. t. aestivum is a heterogenous species. montecchi and sarasini (2000) mention the results of studies indicating 18 various clones or breeds of this species but none of them reached the species level. the polish collections of t. aestivum present great variability, as well. the study has just started. the collections from three localities contain fruit bodies of slightly different character, what allow considering them to be close to tuber uncinatum chat. according to riousset et al. (2001). ecology. the fruit bodies were found in association with fagus sylvatica, quercus robur, q. petraea and variety of other deciduous trees on calcareous soil. the collections come from three uplands, mostly from the lower parts of the hills. the species was found in places of heavy human impact at the forest paths and tourist trails, in fig. 1 146 m. ławrynowicz et al. the vicinity of houses or in the area facing a new arrangement of roads. it means that the occurrence of t. aestivum is seriously endangered (pl. i 1, 3). distribution. t. aestivum is considered to be the most widespread species among edible truffles in europe, occurring from mediterranean zone to south scandinavia (veden et al. 2001). lubelska (1954) gives six localities of the species in poland mainly on the basis of popular literature. recent collections of t. aestivum are indicated in the figure 1. localities de 59 – wy ż y n a pr z e d b o r s k a , przedborski landscape park, dobromierz by przedbórz, fagus sylvatica, aug. 2007, leg. tk, lod 22006. de 84 – wy ż y n a k r a k o w s k o c z ę s t o c h o w s k a , jurajski landscape park, mstów, jul. 2007, quercus petraea, fagus sylvatica, leg. mł, lod 22014. res. zielona góra, fagus sylvatica, carpinus betulus, quercus robur, jul. 2007, leg mł, lod 22015. de 95 – wy ż y n a k r a k o w s k o c z ę s t o c h o w s k a , jurajski landscape park, lud-ludwinów by złoty potok, fagus sylvatica, jul. 2008, leg. tk, lod 22008. df 49 – wy ż y n a k r a k o w s k o c z ę s t o c h o w s k a , złożeniec by smoleń, fagus sylvatica, quercus petraea, carpinus betulus, jul. 2007, leg. tk, lod 22010. ee 94 – wy ż y n a k i e l e c k o s a n d o m i e r s k a , dębina by lisów, carpinus betulus, quercus petraea, pinus sylvestris, sept., nov. 2007, leg. mf, lod 22002. tuber aestivum vittad. forma uncinatum (chatin) montecchi et borelli 1990 = t. uncinatum chatin 1887. fruit bodies similar to those of t. aestivum vittad. the differences are in smaller warts on the surface, darker colour of gleba, more intensive odour, spores ornamented with more distinct reticulum with meshes up to 4 μm high, curved in the upper part. the last feature was observed only in some spores. on the basis of three collections we concluded that t. aestivum forma uncinatum is a late autumn form of t. aestivum adapted to more humid sites (pl. i 2, 4, 5, 6). remarks. recent collections of t. aestivum present a great taxonomical variability of this species. the most distinctly separated group is that showing the features of t. aestivum forma uncinatum. continuation of collecting and taxonomical studies may bring additional arguments to verify our point of view in this question. ecology. in humus-clay soil in calcareous area. under carpinus betulus, acer pseudoplatanus, quercus petrea, corylus avellana and other trees. on hills up to 290 m a.s.l., but usually in the lower parts; autumn and winter mounts; in great quantities: more than a hundred of carpophores were collected in three localities. distribution. the localities are indicated in figure 1. the occurrence is endangered because of forest management practices and penetration by people from adjacent villages. the whole area should be protected. localities ee 94 – wy ż y n a k i e l e c k o s a n d o m i e r s k a , piotrkowice, acer pseudoplatanus, quercus petraea, carpinus betulus, betula pendula, dec. 2007, leg. mf, lod 22004. załatwie by lisów, carpinus betulus, jan. 2008, leg. mf, lod 22003. małolipie by skrzeczyce, carpinus betulus, prunus spinosa, rosa canina. jan. 2008, leg. mf, lod 22005. occurrence of black truffles 147 tuber mesentericum vittadini 1831 = tuber bituminatum berkeley et broom 1851. fruit bodies globose or subglobose with basal or side cavity 1-5 cm diam., firm, black or blackish blue when fresh, becoming blackish gray when dried. peridium warted with rather small, obtuse warts 3-4 mm across. gleba white when fresh becoming blue gray, yellowish to light brown when dried, marbled with whitish veins. odour distinct, pleasant when not too much concentrated. asci 60-100 × 55-80 μm, broadly ellipsoid, sometimes subglobose, with a short stalk, 1-5 spored. ascospores 27-53 × 20-35 μm, ellipsoid or subglobose, hyaline when young, yellowish brown at maturity, ornamented with a coarse reticulum 3-5 μm high, usually 3-5 across the width of spore (pl. ii 3, 4). remarks. tuber mesentericum has been regarded as synonym of t. aestivum by several authors (e.g. pegler et al. 1993), but the others confirmed it as separate species (knapp 1951; ceruti 1960; montecchi, sarasini 2000; riousset et al. 2001; kers 2003). t. mesentericum was distinguished from t. aestivum by having smaller fruit bodies, ornamented with smaller warts, obtuse or depressed at the apex, black colour of surface with black-blue reflex becoming black gray when dried, having cavity at the base or on side of carpophores. the spores of t. mesentericum are larger and ornamented with more complete reticulum than these of t. aestivum. examination of rich materials collected by the authors supports the separation of these two species (pl. ii 2). fig. 2 148 m. ławrynowicz et al. ecology. in humus-sandy soil, clay mixed with small calcareous stones. under fagus sylvatica, carpinus betulus, corylus avellana, quercus robur, q. petraea. t. mesentericum was found in different sites as t. aestivum. even when the same locality has been stated for both species, they do not grow together but in different sites separated by distance of at least 50 m. distribution. this is the only species of black truffles monitored in two localities since 1981 and 1997, respectively. together with recent collections, it is known from five localities indicated in figure 2. exact distribution of the species in europe is difficult to determine because t. mesentericum and t. aestivum had been treated as one taxon for a long time. localities de 84 – wy ż y n a k r a k o w s k o c z ę s t o c h o w s k a , jurajski landscape park, res. zielona góra (ławrynowicz 1988, 1990), jul.2007,leg. mł, lod 22017 wancerzów (ławrynowicz 1999), jul. 2007, leg. mł, lod 22009. df 37 − wy ż y n a k r a k o w s k o c z ę s t o c h o w s k a , jurajski landscape park, hutki-kanki by olkusz, jul. 2007, leg. tk, lod 22017. ee 72 – wy ż y n a k i e l e c k o s a n d o m i e r s k a , res. milechowy, carpinus betu�betulus, corylus avellana, quercus petraea, pinus sylvestris, cornus sp., sept. 2007, leg. mf, lod 22016. tuber bellonae quélet 1887 = tuber bituminatum berkeley et broome var. sphaerosporum ferry de la bellone. fruit bodies irregularly globose, 3-5 cm in diam., with depressions or small cavity; black on surface. peridium covered with pyramidal, obtuse warts. gleba light, becoming yellowish brown. odour intensive, not very pleasant when concentrated. asci ovoid 77-115 × 77-95 μm with 1-6 spores. spores globose or subglobose, 22.555 μm in diam.; ornamented with fine reticulum 5-7 μm high; alveoli 4-6 angled, 5-10 μm long (pl. ii 5, 6). remarks. according to some authors (eg. pacioni & fantini 1997; riousset et al. 2001) t. bellonae posesses the features intermediate between t. mesentericum and t. aestivum forma uncinatum. ecology. in humus-clay or clay soils mixed with small calcareous stones, usually covered with thin litter layer. tuber bellonae has been found under quercus petraea, corylus avellana and fagus sylvatica on calcareous soils (pl. ii 1). distribution. as the species has not been distinguished as a separate taxon by many authors for a long time, its real distribution is impossible to determine. the collections comes from three localities in poland indicated in the figure 3. localities de 59 – wy ż y n a pr z e d b o r s k a , przedborski landscape park, dobromierz by przedbórz, fagus sylvatica, aug. 2007; leg. tk, lod 22011. de 95 – wy ż y n a k r a k o w s k o c z ę s t o c h o w s k a , jurajski landscape park, ludwinów on złoty potok, fagus sylvatica, jul. 2008, leg. tk, lod 22007. ee 82 – wy ż y n a k i e l e c k o s a n d o m i e r s k a , grzywy korzeczkowskie, quercus petraea, corylus avellana, aug., nov. 2007, leg. mf, lod 22001. occurrence of black truffles 149 general remarks and conclusions increasing interest in hypogeous fungi in poland results in discovering of sev-ing interest in hypogeous fungi in poland results in discovering of several new localities and gathering big collections of carpophores, among them black truffles. this activity is of a great value for research and practice. the knowledge of occurrence of black truffles in natural habitats and of its distribution could help to better recognize the ecological requirements of particular truffle species. the results of taxonomical, ecological and chorological analyses are concluded in the identification of four taxa: tuber bellonae quél., t. mesentericum vittad., t. aestivum vittad., and t. aestivum vittad. forma uncinatum (chat.) montecchi et borelli. the collected material, consisting of more than 200 fruit bodies, came from 13 localities in calcareous uplands in the south of poland. some collections were also signalised by hilszczańska et al. (2008). in the present paper, tuber bellonae is reported from poland for the first time. t. mesentericum has been found and continuously observed since 1981; t. aestivum was mentioned in the literature in 19. century in 6 localities in different parts of poland (lubelska 1954). no exsiccates of the latter species coming from these sites were found neither in polish nor in foreign herbaria, therefore it is impossible to know its exact taxonomical interpretation. the recent collections of fresh carpophores reveal intraspecific variation of tuber aestivum, enabling us to distinguish a group of uncinatum in the rank of a form. the characteristic features of this taxon are attributed to the collections coming from the most northeastern localities and autumn as well as winter period of occurrence. fig. 3 150 m. ławrynowicz et al. through examination of tens of carpophores revealed that part of them do not reach the maturity during the whole vegetation season. on the other hand it comes clear that the production of black truffle carpophores in poland in the last years increases. moreover, it seems that fruit bodies of t. mesentericum with cavity and t. bellonae with some depressions in carpophores are better adapted to humid and cold climatic conditions. acknowledgments. many thanks to dr. izabela kałucka for improving the english version of the paper, barbara grzesiak for technical assistance, and an anonymous reviewer for helpful remarks. the study was supported by the polish state committee for scientific research grant no. p04 g 074 25. references alexandrowicz j ., błoński f. 1894 rodzaj tuber mich. trufla. rodzaj choiromyces vittad. piestrak. encyklopedia rolnicza iii: 616. berkeley m. j., broome c. e. 1851. notices of british hypogeous fungi. ann. mag. nat. hist. 2, 7, 183. błoński f. 1888. kilka słów o truflach krajowych i sposobach ich poszukiwania. wszechświat 7: 582–585. caspary r. 1886. trüffeln und trüffelähnliche pilze in preußen. schriften der physik.-ökonom. gesellschaft zu königsberg 27: 177–185. ceruti a. 1960. elaphomycetales et tuberales. (in:) j. bresadola, iconographia mycologica 28, suppl. ii. trento. chatin a. d. 1887. une nouvelle espèce de truffe: tuber uncinatum. c. r. acad. sci. paris, 104, 1132. ferry de la bellone c. 1887. nomenclature et determination des tuberacés et de quelques hypogés récoltés en provence. bsmf 3: 107–110. hilszczańska d., sierota z., palenzona m. 2008. new tuber species fund in poland. mycorrhiza 15: 223–226. kers l. e. 2003. tryfflarna tuber aestivum och t. mesentericum i sverige. svensk botanisk tidskrift 97: (3/4): 157–175. knapp a. 1951. die europäischen hypogaeen-gattungen und ihre gattungstyppen. schweiz. zeitschr. pilzk. 29 (4): 65–96. lubelska b. 1954 (1953). o występowaniu trufli (tuber mich. i choiromyces vitt.) w polsce. – the occurence of the truffles (tuber mich. and choiromyces vitt.) in poland. fragm. flor. geobot. 1: 87–96. ławrynowicz m. 1988. workowce (ascomycetes). jelaniakowe (elaphomyce-tales), truflowe (tuberales). (in:) flora polska. grzyby (mycota) 18. botanical institute, polish academy of sciences, pwn, warszawa-kraków, 161 pp., 17 pls. ławrynowicz m. 1989. chorology of the european hypogeous ascomycetes i. elaphomycetales. acta mycol. 25 (1): 3–41. ławrynowicz m. 1990. chorology of the european hypogeous ascomycetes. ii. tuberales. acta mycol. 26 (1): 7–75. ławrynowicz m. 1999. tuber mesentericum an interesting species of black truffles in poland. acta mycol. 34 (1): 169–172. montecchi a., borelli m. 1990. funghi ipogei raccolti nella localitá vittadiniane: primi risultati. riv. micol. (amb), 33 (3): 278–286. montecchi a., sarasini m. 2000. funghi ipogei d’europa. a m.b., fond centro studi micologici, torino. pacioni g., fantini p. 1997. tuber bellonae un tartufo mediterraneo del complesso tuber aestivum – tuber mesentericum. micol. e veg. mediterr. 12 (1): 15–20. pegler d.n., spooner b.m., young t.w.k. 1993. british truffles. a revision of british hypogeous fungi. royal botanic gardens, kew. quélet l. – quelques espèces critiques ou nouvelles de la flore mycologique de france. bull. de l’assoc. franc. congr. de rouen 12: 498–512. riousset l. et g., chevalier g., bardet m.c. 2001. truffes d’europe et d’chine. inra, paris. vittadini c. 1831. monographia tuberacearum. milano. weden c., ericsson l., danell e. 2001. tryffelnyheter fran gotland (research on tuber aestivum syn. t. uncinatum, and t. mesentericum reported from sweden for the first time). svensk botanisk tidskrift 95: 205–211. occurrence of black truffles 151 występowanie czarnych trufli w polsce s t r e s z c z e n i e grzyby podziemne, a wśród nich trufle są przedmiotem rosnącego zainteresowania z punktu widzenia naukowego i ekonomicznego, z uwagi na wysokie ceny jakie jadalne czarne trufle osiągają na rynkach światowych. centrum występowania czarnych trufli stanowi strefa śródziemnomorska. do strefy umiarkowanej przenikają tylko niektóre gatunki, zaś najdalej, bo do południowej skandynawii i na wyspy brytyjskie sięga trufla letnia tuber aestivum. gatunek ten wykazuje dużą zmienność cech taksonomicznych i bywa rozmaicie traktowany przez poszczególnych autorów zajmujących niekiedy skrajnie różne stanowiska. jedni autorzy (np. pegler i in. 1993) utrzymują t. aestivum jako jeden zbiorowy gatunek, inni zaś (np. riousset i in. 2001) wyróżniają kilka gatunków w grupie t. aestivum, czyli tych trufli, które mają czarne, grubo brodawkowane perydium i zarodniki z ornamentacją siateczkowatą. autorzy niniejszej pracy, dysponując bogatym materiałem w postaci świeżych owocników zebranych w roku 2007 na 13 stanowiskach, podjęli próbę taksonomicznej interpretacji występujących w polsce czarnych trufli. w wyniku analizy makroi mikromorfologicznej, a także ekologicznej i chorologicznej wyróżniono cztery taksony: tuber aestivum vittad. – trufla letnia, t. aestivum vittad. forma uncinatum (chat.) mont. et borelli – trufla letnia forma późna, t. mesentericum vittad. – trufla wgłębiona i t. bellonae quél. – trufla pośrednia. w pracy podano charakterystykę tych taksonów oraz ich rozmieszczenie w polsce. t. bellonae został podany z polski po raz pierwszy. inne gatunki grzybów podziemnych, zebrane podczas badań terenowych będą przedmiotem kolejnych prac. 2014-01-01t11:47:52+0100 polish botanical society some interesting records of cladonia species from the nizina wielkopolska lowland (w poland) daria zarabska1 and christian dolnik2 1natural history collection, faculty of biology, adam mickiewicz university umultowska 89, pl-61-614 poznań, darzarabs12@tlen.pl 2ecology centre, university of kiel, olshausen 40 d 24098 kiel, cdolnik@ecology.uni-kiel.de zarabska d., dolnik ch.: some interesting records of cladonia species from the nizina wielkopolska lowland (w poland). acta mycol. 44 (2): 223–232, 2009. the lichen genus cladonia comprises several similar species which have hardly been recognized in western poland so far. we used thin layer chromatography (tlc) as a simple technique to determine diagnostic lichen substances in morphologically similar cladonia species. during field studies in sandr nowotomyski (western nizina wielkopolska lowland), ten interesting records of cladonia species were made. cladonia novochlorophaea is reported for the first time from this region. our records supplement the knowledge about the distribution of cladonia species both in the investigated region and in poland. all the records are compared with existing literature data from regional floristic inventories and distribution maps from poland. key words: cladonia chlorophaea-group, tlc, sandr nowotomyski, poland introduction the lichen genus cladonia hill ex p. browne comprises several common and widely distributed epigeic species, well known as reindeer or cup lichens. detailed chemical investigations in the 20th century revealed that secondary lichen compounds show a high diversity in cup lichens, and several new species were described on the basis of these lichen substances (e.g., hammer 1995). recently, the importance of secondary lichen metabolites to the taxonomy and species discrimination within the genus cladonia has been confirmed in a first molecular approach by stenroos et al. (2002). though cladonia species can be found in many lichenological reports from different regions of poland (fałtynowicz 2003 and literature cited therein), the information about the species recognizable only by secondary substances is rather acta mycologica vol. 44 (2): 223–232 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 224 d. zarabska and c. dolnik sparse. microcrystallization, thin layer chromatography (tlc), or high pressure liquid chromatography (hplc) are often the only methods to determine specific secondary metabolites (orange et al. 2001), but in poland only a few studies have used these laboratory techniques for the identification of cladonia species so far (e.g., kowalewska et al. 2000, 2008; kowalewska, kukwa 2003, 2007; kowalewska, szok 2004; kukwa 2005a, b; osyczka 2006; oset et al. 2008; syrek, kukwa 2008). although several older papers report epigeic lichens from the nizina wielkopolska lowland, this part of western poland is still underrepresented in the current polish lichen inventory (fałtynowicz, pers. comm.), and therefore every new record from this region is valuable. in this region, attention was paid only to epigeic cladonia species in vegetation surveys of dry grasslands (e.g., celiński, balcerkiewicz 1973) and coniferous forests (e.g., tobolewska, wronówna 1955; długosz 1961; tobolewski 1962, 1963; zarabska 2008a). fragmentary information about epigeic cladonia species can also be found in some phytosociological papers (e.g., kaczyńska 1964; balcerkiewicz, brzeg 1993; balcerkiewicz et al. 1994; bujakiewicz, lisiewska 2003; rakowski 2003). recent lichen inventories carried out in sandr nowotomyski in the western part of wielkopolska lowland in the years 2007 and 2008 revealed several stands of rare and noteworthy cladonia species. some taxa have already been known from the central part of the area in the nowy tomyśl region (krawiec 1938; dziabaszewski 1962; nowacka-zyber 1967; żukiel 1967; bujakiewicz, lisiewska 2003; zarabska 2008a, b). we restrict our study to rare cladonia species and those which require tlc analysis for the identification. our data supplement the information about the distribution range in poland for the following species: cladonia cariosa, c. chlorophaea s.str, c. coccifera, c. crispata, c. grayi, c. merochlorophaea, c. novochlorophaea, c. pyxidata s.str., c. rei and c. subulata. study area localities of cladonia species come from sandr nowotomyski, which is part of the pojezierze poznańskie lakeland (kondracki 2000). according to the geobotanical division of the country, this area belongs to the western part of kraina wielkopolsko-kujawska range (szafer, zarzycki 1972). sandy hills are covered by vegetation of the plant communities vaccinio myrtilli-pinetum sylvestris juraszek 1927 nom. invers. (syn. leucobryo-pinetum matuszkiewicz 1962) and in some places cladinopinetum sylvestris juraszek 1927 nom. invers. (żukiel 1967; medwecka-kornaś 1972; bujakiewicz, lisiewska 2003). these habitats favor the occurrence of epigeic lichen vegetation. some records of cladonia 225 material and methods specimens collected during field work were identified with the help of nowak and tobolewski (1975), purvis and james (1992), and wirth (1995). for the analysis of morphologically similar cladonia species, thin layer chromatography was performed in solvents a and c according to the standard method of culberson and ammann (1979). nomenclature follows fałtynowicz (2003). all samples were collected by the first author, and the specimens are kept in the natural history collection uam poznań (poz). in the following list of species we give precise information about location, detected secondary compounds (if tlc was carried out), about earlier notes in the literature, and about habitats. results altogether, ten species of cladonia are presented here that are rare or have not been recorded in the wielkopolska lowland so far. cladonia cariosa (ach.) spreng.. syst. veg. 4(1): 272 (1827). basionym: lichen cariosus ach., lichenogr. suec. prodr. 198 (1799). this species is readily recognized by longitudinally fissured podetia, which are covered by corticated granules or squamules, and large, clustered, terminal brown apothecia (purvis, james 1992). tobolewski (1988) considered cladonia cariosa as rare species in poland. until now, only a few records from the nizina wielkopolska lowland have been known (krawiec 1933, 1935, 1955; dziabaszewski 1962; tobolewski 1962; nowacka-zyber 1967; kepel 1999). on the local scale it was counted as rare species (dziabaszewski 1962; nowacka-zyber 1967), and was not confirmed during comparison inventories (kepel 1996; glanc 1998). in nowy tomyśl region, it has been reported from only one locality (nowacka-zyber 1967). specimen examined. równina nowotomyska plain, miedzichowo, ca 3 km sw from village, 52°21.10’n, 15°55.55’e, alt. ca 60 m, pine forest, on soil, 24 april 2008. cladonia chlorophaea (flörke ex sommerf.) sprengel, syst. veg. 4: 273 (1827). basionym: cenomyce chlorophaea flörke ex sommerf., suppl. fl. lapp.: 130 (1826). cup lichen with brown apothecia on cup margin. cups gradually tapered to the corticated to granular stalk, inner and outer cup surface usually granular-soredious, sometimes mixed with corticated granules. cladonia chlorophaea s.str. belongs to a group of morphologically similar taxa, and is chemically characterized by the presence of the furmarprotocetraric chemosyndrome (kowalewska et al. 2008). although several localities of this taxon were reported from nizina wielkopolska lowland (krawiec 1933, 1938, 1955; kanarek-drela 1960; długosz 1961; tobolewski 1962; kaczyńska 1964; glanc 1965; nowacka-zyber 1967; żukiel 1967; glanc et al. 1971; kozłowska 1975; balcerkiewicz, brzeg 1993; kepel 1999; rakowski 2003; 226 d. zarabska and c. dolnik bujakiewicz, lisiewska 2003; zarabska 2008a, b), these findings may in fact represent different species as this name was also applied to several other species of the so-called c. chlorophaea-group (fałtynowicz 2003). substances detected by tlc: furmarprotocetraric acid (present in all specimens). specimens examined. równina nowotomyska plain, bolewice, ca 2.2 km se from village, 52°21.40’n, 16°06.54’e , alt. ca 83 m, edge of pine-oak forest near dirt road, on soil, 27 sept. 2007; jastrzębsko stare, ca 2.1 km, 52°17.09’n, 16°05.00’e, alt. ca 70 m, se from village, turf, on soil, 20 sept. 2007. cladonia coccifera (l.) willd., fl. berol. prodr. 361 (1787). basionym: lichen cocciferus l., sp. pl. 1151 (1753). this species is characterized by yellow-green podetia with more or less coarsely corticate-granular or areolate-corticate surface and broad cups, gradually tapering to the base (purvis, james 1992). though nowak and tobolewski (1975) reported cladonia coccifera as widespread in the whole country, in the nizina wielkopolska lowland it has been recorded mostly in the northern part (kozłowska 1975), and only scattered information has been published from other localities in wielkopolska (krawiec 1955; długosz 1961; tobolewski 1962; balcerkiewicz, brzeg 1993; rakowski 2003). nowacka-zyber (1967) considered the species as not so abundant in the nowy tomyśl region. the species is morphologically similar to the very rarely found cladonia borealis (osyczka 2006). the occurrence of barbatic acid in c. borealis is a primary feature to distinguish it from cladonia coccifera (purvis, james 1992). substances detected by tlc: usnic acid (present in all specimens), zeorin (present in all specimens). specimens examined. równina nowotomyska plain, nowa boruja, ca 2.2 km w from village, 52°14.38’n, 16°07.38’e, alt. ca 70 m, edge of a pine forest lined by a dirt road, on soil, 26 sept. 2007; nowy tomyśl, ca 2 km nw from town, 52°20.06’n, 16°06.29’e, alt. ca 95 m, pine forest, on soil, 25 sept. 2007. cladonia crispata (ach.) flot., in wendt, thermen warmbrunn 93 (1839). basionym: baeomyces turbinatus var. crispatus ach., meth. lich. 341 (1803). characterized by the brownish, corticated, irregular branching, podetia terminat-haracterized by the brownish, corticated, irregular branching, podetia terminating in a single perforation surrounded by short spines, and in being pd− (without fumarprotocetraric acid) and uv+ (squamtic acid) (purvis, james 1992). nowak and tobolewski (1975) considered this species as widespread in both lowland and mountainous regions. records in the nizina wielkopolska lowland are rather old and restricted to only a few localities (krawiec 1933; tobolewska, wronówna 1955; długosz 1961; tobolewski 1962, 1963). the species was known mainly from the cladino-pinetum sylvestris forest community (e.g., nowacka-zyber 1967; żukiel 1967). specimens examined. równina nowotomyska plain, błońsko, ca 1.2 km s from village, 52°10.38’n, 16°08.02’e, alt. ca 71 m, mature pine forest, on soil, 07 april 2008; sątopy, ca 1.5 km e from village, 52°19.03’n, 16°13.54’e, alt. ca 101 m, pine forest, on soil, 21 sept. 2007. cladonia grayi merrill ex sandst., clad. exs. no. 1847 (1929). cup lichen with gradually tapering, regular, trumpetor goblet-shaped scyphus, often with proliferations from cup margin. in the upper part, the podetia are covered by granular soredia, while in the lower part, the stalk is corticated, distinctly verruculose and occasionally squamulose (kowalewska et al. 2008). according to kowalewska et al. (2008), cladonia grayi is almost absent in western poland and some records of cladonia 227 partially also in eastern poland; from the nizina wielkopolska lowland it has only been reported by kubiak (2008) so far. in the investigated area the species was found in a clearing of birch and pine forests. substances detected by tlc: grayanic acid (present in all specimens), furmarprotocetraric acid (present in one of two studied specimens). specimens examined. równina nowotomyska plain, chrośnica, ca 2.6 km ne from village, 52°17.08’n, 16°00.40’e, alt. ca 71 m, edge of birch grove, on soil, 27 sept. 2007; jastrzębsko stare, ca 1.8 km nw from village, 52°18.35’n, 16°03.51’e, alt. ca 75 m, pine forest, on soil, 21 sept. 2007. cladonia merochlorophaea asahina, j. jap. bot. 16: 713 (1940). characterized by tall, brownish, greenish-grey podetia with gradually fl ar-haracterized by tall, brownish, greenish-grey podetia with gradually flaring scyphi and areolate-corticate, verruculose, or obscurely sorediate to coarsely granular surface (kowalewska et al. 2008), morphologically indistinguishable from c. novochlorophaea. several localities of c. merochlorophaea were discovered in the western poland by kowalewska et al. (2008). in wielkopolska it was previously noted only by rakowski (2003) and zarabska (2008a, b), but seems to be the most common member of the cladonia chlorophaea-group in this region. in accordance with results of kowalewska et al. (2008), c. merochlorophaea was mainly recorded on humus-rich soil. substances detected by tlc: merochlorophaeic and 4-o-methylcryptochlorophaeic acid (present in all specimens), furmarprotocetraric acid (present in four of seven studied specimens). specimens examined. równina nowotomyska plain, nowa boruja, ca 2.2 km w from village, 52°14.38’n, 16°07.38’e, alt. ca 70 m, edge of pine forest lined by a dirt road, on soil, 26 sept. 2007; chrośnica, ca 1.0 km se from village, 52°15.47’n, 16°00.51’e, alt. ca 80 m, on sunny clearing in pine forest, on soil, 27 sept. 2007; chrośnica, ca 2.6 km ne from village, 52°17.08’n, 16°00.40’e, 27 sept. 2007, alt. ca 71 m, edge of birch grove, on soil; jastrzębsko stare, ca 2.1 km se from village, 52°17.09’n, 16°05.00’e, alt. ca 70 m, turf, on soil, 20 sept. 2007; jastrzębsko stare, ca 1.8 km nw from village, 52°18.37’n, 16°02.49’e, alt. ca 75 m, pine forest, on humus, 19 sept. 2007; miedzichowo, ca 3 km sw from village, 52°21.10’n, 15°55.55’e, alt. ca 60 m, pine forest, on soil, 24 april 2008; bolewice, ca 2.7 km se from village, 52°22.48’n, 16°06.54’e, alt. ca 80 m, pine forest, on soil, 2 oct. 2007. cladonia novochlorophaea (sipman) brodo ahti, canad. j. bot. 74: 1167 (1996). basionym: cladonia merochlorophaea asahina var. novochlorophaea sipman, acta bot. neerl. 22: 496 (1973). according to kowalewska et al. (2008), this cup lichen is morphologically indistinguishable from c. merochlorophaea and was only recently discovered in poland (kowalewska, kukwa 2001). until now, it was rarely noted in the country and was recorded mainly from the northern part (kowalewska et al. 2008). it can be distinguished from c. merochlorophaea by the presence of homosekikaic and sekikaic acids. here, the species is reported for the first time for the nizina wielkopolska lowland. substances detected by tlc: homosekikaic acid and sekikaic acid (present in all specimens), furmarprotocetraric acid (present in one of two studied specimens). specimens examined. równina nowotomyska plain, jastrzębsko stare, ca 2.1 km se from village, 52°17.09’n, 16°05.00’e, alt. ca 70 m, turf, on soil, 20 sept. 2007; bolewice, ca 2.2 km se from village, 52°21.40’n, 16°06.54’e, alt. 90 m, pine-oak forest, on soil, 27 sept. 2007. 228 d. zarabska and c. dolnik cladonia pyxidata (l.) hoffm. s.str., deutschl. fl. 2: 121 (1796). basionym: lichen pyxidatus l., spec. pl. 2: 1151 (1753). syn. c. callosa delise, c. neglecta (flörke) spreng. gradually flaring from the base to the top, greenish grey to brownish podetia are covered with an irregular areolate cortex (kowalewska et al. 2008). cladonia pyxidata s.str. is mainly found in the south of poland. from the western part of the country only scattered localities are known (kowalewska et al. 2008). this species was often recorded in floristic studies of the nizina wielkopolska lowland (krawiec 1933; dziabaszewski 1962; tobolewski 1963; glanc 1965, 1969, 1998; nowackazyber 1967; żukiel 1967; kozłowska 1975; brzeg 1981; balcerkiewicz , brzeg 1993; balcerkiewicz et al. 1994; rusińska 1999; rakowski 2003), but numerous records may have been mistaken for morphologically similar species, e.g. cladonia merochlorophaea, c. chlorophaea, c. grayi, or c. monomorpha. cladonia pyxidata s.str. is mentioned only in an article of bujakiewicz, lisiewska (2003), who recorded this lichen also in the nowy tomyśl region. substances detected by tlc: furmarprotocetraric acid (present in all specimens). specimens examined. równina nowotomyska plain, jastrzębsko stare, ca 2.1 km se from village, 52°17.09’n, 16°05.00’e, alt. ca 70 m, turf, on soil, 20 sept. 2007; nowy tomyśl, ca 2 km nw from town, pine forest, on soil, alt. ca 95 m, 52°20.06’n, 16°06.29’e, 25 sept. 2007. cladonia rei schaer., lich. helv. spicil.: 34 (1823). syn. c. nemoxyna (ach.) arnold. characterized by long, granular-sorediate to farinose podetia with or without narrow cups with short proliferations, by often prominent apothecia, or by stout podetia with short anisodiametric branching patterns. this species can be confused with the morphologically similar c. glauca and c. subulata (kukwa 2005a). in poland, cladonia rei is common in the eastern part of the country and has been recorded from the wielkopolska lowland only once (syrek, kukwa 2008). lately, the taxonomical status of c. rei has been under discussion (spier, aptroot 2007; syrek, kukwa 2008); however, most recent molecular studies by us (dolnik, beck and zarabska subm.) proved the distinctness of c. rei as a separate species, which is not closely related to c. subulata. substances detected by tlc: homosekikaic acid (present in all specimens), furmarprotocetraric acid (present in two of four studied specimens). specimens examined. równina nowotomyska plain: jabłonna, ca 2.2 km n from village, 52°13.32’n, 16°12.19’e, alt. ca 85 m, pine forest, on soil, 17 sept. 2008; szarki, ca 2.5 km s from village, 52°15.30’n, 16°04.21’e, on the edge of a pine forest, on soil, alt. ca 70 m, 25 sept. 2008; chrośnica, ca 2.6 km ne from village, 52°17.08’n, 16°00.40’e, alt. ca 71 m, edge of birch grove, on soil, 27 sept. 2007; jastrzębsko stare, ca 2.1 km se from village, 52°18.35’n, 16°03.51’e, alt. ca 70 m, turf, on soil, 20 sept. 2007. cladonia subulata (l.) weber in f. h. wigg., prim. fl. holsat. 90 (1780). basionym: lichen subulatus l. sp. pl. 1153 (1753). syn. c. cornutoradiata (coem.) zopf, c. fimbriata var. cornutoradiata coem. characterized by long subulate podetia with a granular to farinose sorediate sur-haracterized by long subulate podetia with a granular to farinose sorediate surface, often with an antler-like branching pattern, and by prominent long-stalked cups with long and unequal proliferations narrowing to an acute tip. in contrast to cladonia rei, this species was often recorded in the wielkopolska area (krawiec 1930, 1933, 1938, 1955; tobolewska, wronówna 1955; dziabaszewski 1962; tobolewski some records of cladonia 229 1963; glanc 1969, 1998; glanc et al. 1971; kozłowska 1975; tobolewski, kupczyk 1977; balcerkiewicz, brzeg 1993; brzeg, pawlak 1998; kepel 1996, 1999; rakowski 2003), but misidentifications through confusion with furmarprotocetraric acid containing morphs of c. rei cannot be ruled out completely. cladonia subulata was recorded in six new localities in the western part of wielkopolska. these were also the first recordings of this species for sandr nowotomyski. substances detected by tlc: furmarprotocetraric acid (present in all specimens). specimens examined. równina nowotomyska plain, komorówko, ca 1.6 km sw from village, 52°10.42’n, 16°10.19’e, pine forest, on soil, alt. ca 78 m, 07 april 2008; jabłonna, ca 2.2 km n from village, 52°13.32’n, 16°12.19’e, alt. ca 85 m, on soil, 17 sept. 2008; chrośnica, ca 2.6 km ne from village, birch forest, 52°17.08’n, 16°00.40’e, on soil, alt. ca 71 m; jastrzębsko stare, ca 2.1 km se from village, 52°18.35’n, 16°03.51’e, alt. ca 70 m, turf, on soil, 20 sept. 2007; sękowo, ca 0.9 km nw from village, 52°18.40’n, 16°04.57’e, alt. ca 71 m, pine forest, on soil, 20 sept. 2007; sątopy, ca 1.5 km e from village, 52°19.03’n, 16°13.54’e, alt. ca 101 m, pine forest, on soil, 21 sept. 2007. conclusion western poland, especially sandy areas with inland dunes, comprises an interesting epigeic lichen flora with several cladonia species. the use of tlc for the identification of lichens collected in the wielkopolska region revealed several records of rare species, which have been absent in older sources of poland. the previously poorly recognized species cladonia merochlorophaea turned out to be the most common member of the cladonia chlorophaea-group in the region. acknowledgements. we thank karol latowski (poznań), and wiesław fałtynowicz (wrocław), and anonymous reviewers for helpful suggestions on an earlier draft of the manuscript, and aiko huckauf (kiel) for improving the language. references balcerkiewicz s., brzeg a. 1993. wrzosowiska przydrożne w kompleksie leśnym borów skwierzyńskich. bad. fizjogr. pol. zach., ser. b, 42: 105–127. balcerkiewicz s., brzeg a., kasprowicz m. 1994. szata roślinna rezerwatu „nadwarciański bór sosnowy” w wielkopolskim parku narodowego. bad. fizjogr. pol. zach., ser. b, 43: 51–83. brzeg a., pawlak g. 1998. materiały do znajomości zbiorowisk związku onopordion acanthii br.-bl. 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(eds). 1972. szata roślinna polski. pwn, warszawa. tobolewska r., wronówna i. 1955. bory chrobotkowe w okolicach wronek i sierakowa na wydmach międzyrzecza warciańsko-noteckiego. spraw. ptpn 1: 315–317. tobolewski z. 1962. materiały do flory porostów północno-zachodniej polski. fragm. florist. geobot. 8: 67–80. tobolewski z. 1963. materiały do znajomości borów chrobotkowych północno-zachodniej polski. bad. fizjogr. pol. zach. 12: 193−211. tobolewski z. 1988. porosty (lichenes). 9. atlas rozmieszczenia roślin zarodnikowych w polsce, ser. iii. pwn, warszawa-poznań. tobolewski z., kupczyk b. 1977. porosty (lichenes). 4. atlas rozmieszczenia roślin zarodnikowych w polsce, ser. iii. pwn, warszawa –poznań. wirth v. 1995. die flechten baden-w�rttembergs. verlag eugen ulmer, stuttgart. zarabska d. 2008a. potencjalne bogactwo gatunkowe porostów projektowanego parku krajobrazowego sandr nowotomyski na przykładzie naziemnych chrobotków cladonia sp. obszaru „glińskie góry”. biul. park. krajobr. wielkopolski 14 (16): 66–72. zarabska d. 2008b. porosty jako bioindykatory zanieczyszczenia powietrza w okolicach nowego tomyśla. bad. fizjogr. pol. zach., ser. b, 57: 109–121. żukiel z. 1967. bory sosnowe okolic nowego tomyśla. katedra systematyki i geografii roślin uam, poznań (mscr.). 232 d. zarabska and c. dolnik interesujące notowania cladonia spp. z niziny wielkopolskiej (polska zachodnia) streszczenie rodzaj cladonia zawiera kilka podobnych taksonomicznie gatunków, które były dość rzadko podawane z obszaru polski zachodniej. w trakcie identyfikacji gatunków wykorzystaliśmy metodę chromatografii cienkowarstwowej (tlc) pozwalającą na określenie substancji porostowych mających charakter diagnostyczny u podobnych pod względem morfologicznym gatunków rodzaju cladonia. podczas badań terenowych na sandrze nowotomyskim (zachodnia wielkopolska) odnotowano interesujące gatunki rodzaju cladonia (c. cariosa, c. chlorophaea, c. coccifera, c. crispata, c. grayi, c. merochlorophaea, c. novochlorophaea, c. pyxidata s.str., c. rei i c. subulata). cladonia novochlorophaea została stwierdzona po raz pierwszy w wielkopolsce. podane informacje pozwolą uzupełnić wiedzę na temat rozmieszczenia chrobotków zarówno w objętym inwentaryzacją regionie, jak i w polsce. przy analizach występowania wyszczególnionych taksonów wykorzystano informacje zawarte w danych literaturowych i na mapach rozmieszczenia tych gatunków w polsce. 2014-01-01t11:49:35+0100 polish botanical society natural substrata for corticioid fungi eugene o. yurchenko laboratory of mycology, v.f. kuprevich institute of experimental botany akademichnaya 27, by 220072, minsk fungi@biobel.bas net.by y u r c h e n k o e . o . : natural substrata for corticioid fungi. acta mycol. 42 (1): 113 124, 2006. the paper reviews the types of substrata inhabited by non poroid resupinate homobasidiomycetes in situ in global scale with both examples from literature sources and from observations on belarus corticioid fungi biota. the groups of organic world colonized by corticioid basidiomata and vegetative mycelium are arboreous, semi arboreous, and herbaceous vascular plants, bryophyta, epiphytic coccoid algae, lichenized and non lichenized fungi, and occasionally myxomycetes and invertebrates. the fungi occur on living, dying, and dead on all decay stages parts of organisms. besides, the fungi are known on soil, humus, stones, artificial inorganic and synthetic materials and dung. key words: bryophyta, corticiaceae, herbs, lichens, litter, woody plants introduction corticioid fungi (homobasidiomycetes) is an artificial union of life forms, the assembling of which is based mostly on basidioma morphological organization similarity. our definition of corticioid basidiomata sensu lato includes the types of fructifications ranged from totally resupinate to effuse-reflexed with rather wide effused part in most cases, and from loose arachnoid to crustose and membranaceous in consistency; the hymenophore shape varies from smooth to almost poroid (reticulate or irpicoid) and long-toothed. some mycologists still use for them rather convenient name corticiaceae s. l. according to the recent molecular phylogeny-based systems the species of corticioid fungi are distributed among the orders agaricales, boletales, cantharellales, ceratobasidiales, hymenochaetales, phallales, polyporales, russulales, stereales, thelephorales (k i r k et al. 2001) or between athelioid, bolete, cantharelloid, corticioid, euagarics, gloeophyllum, gomphoid-phalloid, hymenochaetoid, polyporoid, russuloid, thelephoroid, and trechisporoid clades sensu b i n d e r et al. (2005). corticioid fungi are a very significant group of wood decay organisms. the type of nutrition for no less than 80% of species is lignin degradation that means ultimately the transformation of dead wood into humus substances, and ca 5% of species cause acta mycologica vol. 41 (1): 113-124 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 114 e. o. yurchenko cellulose decay. besides, there are mycorrhiza-forming species and a part of species which are able to decay different soft plant debris in litter. the facts of corticiaceae s. l. growth on non-wood substrata were documented by different authors, but still the data are strongly dispersed. the term substratum according to h a w k s w o r t h et al. (1995: 445) means ‘the material on which an organism is growing or to which it is attached’. in the first case we tell about nutritive substratum, in the second – about attachment substratum. since fungal thallus contacts with great number of different particles we use the term in more strict sense – the body to which the fungus attach by its basidioma basal part or vegetative mycelium, or in which fungal hyphae penetrate. thus the cases of mechanical embracing of outer particles by upper layers of big fruitbodies are excluded (for example, we observed the embracing of living fragaria vesca l. tendrils by growing chondrostereum purpureum (pers.) pouzar, msk 6519). different substrata for corticiaceae s. l. in situ are discussed in sections below. the specimens examined are kept in v.f. kuprevich institute of experimental botany herbarium, section “fungi” (msk-f). photographs were made by olympus camedia c-5060 digital camera via olympus sz61 stereomicroscope and olympus bx51 microscope; and also by nikon coolpix 4500 digital camera directly or via nikon eclipse e200 microscope with objective magnification 4×. microscopic preparations for passing light were made in 5% koh solution. the correct names of fungi collected in belarus follow cortbase vers. 2 (p a r m a s t o et al. 2004) and k õ l j a l g (1996); in referring to publications the original names are cited, with the orthography checked via cortbase. vascular plants corticiaceae s. l. are known on all kinds of woody plants – angiosperms, gymnosperms, and arboreous ferns (tubulicium vermiculare (wakef.) boidin & gilles and t. dussii (pat.) oberw. fide d o m a ń s k i 1992). main substrata are tree trunks and branches. rather big number of species occur regularly or occasionally on processed or man-transformed wood (timbers, fences, woodwork, chips), but commonly exposed to open air. for instance, a a n d s t a d and r y v a r d e n (1987) listed 82 species on wood fences in norway. the number of species growing indoor is much less – e.g. b o n d a r t s e v (1948) reported 9 house fungi collected in big city. a number of species are known from small shrubs – arctostaphylos, chamaecytisus, cistus, duschekia, erica, genista, ledum, pentaphylloides, sarothamnus, thymus (including strictly specialized coronicium thymicola (bourdot & galzin) jülich), vaccinium (e.g. two species described only from this genus – laeticorticium efibulatum m.j. larsen & nakasone and peniophora sphaerocystidiata burds. & nakasone), and woody lianas, like actinidia and vitis (j ü l i c h 1984; d o m a ń s k i 1988, 1991, 1992; k õ l j a l g 1996). among small shrubs rather rich species collection is reported on calluna stems and twigs: acanthobasidium norvegicum (j. erikss. & ryvarden) boidin, acanthophysium apricans (bourdot) g. cunn., corticium macrosporopsis jülich, hyphodontia hastata (litsch.) j. erikss., phanerochaete ericina (bourdot) j. erikss. & ryvarden, ph. martelliana (bres.) j. erikss. & ryvarden, sistotrema dennisii malençon – on living stem bases, stereum rameale (pers.) fr., trechispora natural substrata for corticioid fungi 115 praefocata (bourdot & galzin) liberta (d o m a ń s k i 1988, 1991, 1992). peniophora halimi boidin & lanquetin and radulomyces rickii (bres.) m.p. christ. inhabit stems of shrubby halophyte atriplex halimus l. (d o m a ń s k i 1991). among our collections there are amphinema byssoides (pers.) j. erikss. (msk 5730), piloderma fallax (liberta) stalpers (msk 6098), and tomentella terrestis (berk. & broome) m.j. larsen (msk 6299) on living and dead vaccinium myrtillus l. stems, immersed in litter. some species also occur on semi-arboreous plants, i.e. those with every year dying younger sprouts. on rubus idaeus l. and r. nessensis w. hall dead stems we collected in belarus ceratobasidium cornigerum (bourdot) d.p. rogers, peniophora cinerea (pers.) cooke, p. incarnata (pers.) p. karst., phanerochaete sanguinea (fr.) pouzar, phlebiella sulphurea (pers.) ginns & lefebvre, radulomyces confluens (fr.) m.p. christ., and tomentella spp. other saprobic species known on rubus are acanthobasidium norvegicum, corticium macrosporopsis jülich, peniophora meridionalis boidin, phanerochaete tuberculata (p. karst.) parmasto, sistotrema pteriphilum k.h. larss. & hjortstam, tomentella coerulea (bres.) höhn. & litsch., and t. ellisii (sacc.) jülich & stalpers (d o m a ń s k i 1988, 1991, 1992; k õ l j a l g 1996). litschauerella clematitis (bourdot & galzin) j. erikss. & ryvarden, peniophora pseudoversicolor boidin, and raduomyces rickii were published from clematis (d o m a ń s k i 1991). partly lignified arundinaria and sasa serve as hosts for acanthobasidium phragmitis boidin & al., cyphellathelia pezizoidea (ellis & everh.) jülich, and tomentella sublilacina (ellis & holw.) wakef. (d o m a ń s k i 1988; k õ l j a l g 1996). several corticioid fungi are known from succulent perennial plants, e.g. athelia decipiens (höhn. & litsch.) j. erikss., hyphoderma fouquieriae nakasone & gilb., peniophora tamaricicola boidin & malençon, phanerochaete omnivorum (shear.) burds. & nakasone, ph. tuberculata on cacti carnegiea and opuntia, crustoderma opuntiae nakasone & gilb. and uncobasidium calongei (tellería) hjortstam & tellería described only from opuntia, and laetisaria agaves burds. & gilb. on agave leaves (n a k a s o n e , g i l b e r t s o n 1978; d o m a ń s k i 1988, 1992). the degree of specialization to vascular hosts varies for corticioid fungi species in wide limits. in belarus only eleven species are still found to be strictly specialized to single plant and regularly recorded on it: amylostereum laevigatum (fr.) boidin (on juniperus communis l.), corticium quercicola jülich (on quercus robur l.), meruliopsis taxicola (pers.) bondartsev (on pinus sylvestris l.), peniophora laeta (fr.) donk (on carpinus betulus l.), p. limitata (chaillet.) cooke (on fraxinus excelsior l.), p. pini (schleich. & dc.) boidin (on pinus sylvestris), p. polygonia (pers.) bourdot & galzin (on populus tremula l.), p. rufomarginata (pers.) litsch. (on tilia cordata mill.), phlebiella pseudotsugae (burt) k.h. larss. & hjortstam (on pinus sylvestris), punctularia strigosozonata (schwein.) p.h.b. talbot (on populus tremula), sistotremastrum suecicum litsch. (on pinus sylvestris). it shows uneven distribution of highly specialized fungi over the genera, with big per cent among peniophora species. several species are genera-specialized, e.g. peniophora erikssonii boidin recorded on alnus glutinosa (l.) gaertn. and a. incana (l.) moench, and vuilleminia alni boidin, lanquetin & gilles on the same two hosts. concerning peniophora pini mentioned above, outside of belarus it was collected on other pinus species; we examined specimens from p. kochiana klotzsch (russian caucasus, msk 6566) and p. nigra arnold (čech republic, collected by w. wojewoda, kram-f 53637). 116 e. o. yurchenko the other type of specialization is the occurrence of a fungus frequently on one host and very rarely on more 1–2 hosts – like peniophora quercina (pers.) cooke frequently on quercus robur and once on corylus avellana l. (msk 4562) in our collections. a clear distinguishing group of fungi when consider substratum specialization are inhabitants of coniferous species only. in belarus the species still recorded on picea abies (l.) karst. and p. sylvestris only are amylostereum chailletii (pers.) boidin, hyphodontia alutacea (fr.) j. erikss., phlebiopsis gigantea (fr.) jülich, pseudomerulius aureus (fr.) jülich, and serpula himantioides (fr.) p. karst. contrary, several species have very wide range of hosts, e.g. a good represented in msk-f herbarium peniophora cinerea (211 specimens from belarus) was recorded in belarus on 44 vascular hosts: acer japonicum thunb., a. platanoides l., aesculus hippocastanum l., alnus glutinosa, a. incana, amelanchier ovalis medik., betula pendula roth, b. pubescens ehrh., carpinus betulus, cerasus vulgaris mill., chamaenerion angustifolium (l.) scop., corylus avellana, fagus sylvatica l., frangula alnus mill., fraxinus excelsior, fraxinus sp., ligustrum vulgare l., lonicera xylosteum l., malus domestica borkh., m. domestica×prunifolia, m. ×prunifolia (willd.) borkh., m. sylvestris mill., padus avium mill., populus tremula, prunus domestica l., pyrus communis l., p. domestica medik., quercus borealis michx. f., q. robur, ribes nigrum l., r. rubrum l., rosa cf. subcanina (christ) dalla torre & sarnth., rosa sp., rubus nessensis, salix caprea l., s. cinerea l., sorbus aria (l.) crantz, s. aucuparia l., s. hybrida l., syringa vulgaris l., tilia cordata, plus on debris of three unidentified herbaceous species near wood. such rich list of substrata is explained also by the wide ecological amplitude in respect to biotope types. species like peniophora cinerea and p. incarnata commonly found in ornamental woody plantations and dendrological collections, which extends their opportunities to colonize exots. though, we have not collections of p. cinerea from gymnosperms. similar multi-host behavior was observed for radulomyces confluens, collected on amelanchier ovalis, armeniaca vulgaris lam., caragana arborescens lam., cerasus vulgaris, crataegus sp., fraxinus excelsior, malus domestica, m. domestica×prunifolia, m. ×prunifolia, m. sylvestris, padus avium, picea abies, prunus divaricata ledeb., p. domestica, pyrus communis, p. domestica, quercus robur, ribes nigrum, rubus idaeus, sorbus aucuparia. in respect to wood decomposition the fungi inhabit it on all decay stages from still living and starting to decay to almost transformed into humus. still not clear studied subject is the relation of fungi to living, dying, and recently dead tissues of vascular plants as substratum. not so easy to delimit the border between biotrophy, necrotrophy, and saprotrophy for wood-inhabiting fungi. our purposeful search of corticiaceae s. l. on living parts of trees has shown that the list of species is bigger than it was documented by previous workers. the most often event is basidiomata patches found on trunk wound calluses, i.e. roller-shaped excrescences around former wounds, covered by thin bark with rather chlorophyll-rich cells in cortex parenchyma. the fungi which were found in former wounds simultaneously on dead wood and on surrounding living tissues in belarus are byssomerulius corium (pers.) parmasto, coniophora puteana (schumach.) p. karst., cylindrobasidium evolvens (fr.) jülich, hymenochaete tabacina (sowerby) lév., hyphodontia crustosa (pers.) j. erikss., h. sambuci (pers.) j. erikss., hypochnicium bombycinum (sommerf.) j. erikss., lagarobasidium detriticum (bourdot) jülich, peniophora cinerea, p. incarnata (fig. 1), phanerochaete sordida (p. karst.) j. erikss. & ryvarden, phle natural substrata for corticioid fungi 117 bia aurea (fr.) nakasone, ph. tremellosa (schrad.) nakasone & burds., steccherinum fimbriatum (pers.) j. erikss., stereum hirsutum (willd.) gray. several species were also collected from bark on the borded of dead and living tissues of trunk and branches: hyphoderma mutatum (peck) donk, h. setigerum (fr.) donk, peniophora nuda (fr.) bres., phlebia radiata fr., radulomyces confluens. besides trunks and branches, corticioid fungi are found on all kinds of woody plant organs, including roots, tendrils (fig. 2), and fallen parts – bark pieces, leaf plates and petioles, fruits, cones. on fallen female cones of picea and pinus we collected common litter fungi amphinema byssoides, athelia epiphylla complex, phlebiella sulphurea, and more rarely tylospora asterophora (bonord.) donk; ceratobasidium cornigerum was found on cone peduncle. besides, tylospora fibrillosa (burt) donk was collected on fallen male cones of pinus sylvestris (fig. 3). several species are parasitic on tree leaves – ceratobasidium ramicola c.c. tu, d.a. roberts & kimbr., corticium stevensii burt, and koleroga noxia donk (d o m a ń s k i 1991). ceratobasidium cornigerum-like species, published under the name ceratobasidium aff. ramicola c.c. tu, d.a. roberts & kimbr. (yurchenko 2003), was found fructifying on living juniperus communis needles. according to j ü l i c h (1984) 20 species inhabit fallen angiosperm leaves, among them six species of athelia and three of byssocorticium; dacryobasidium lutescens (j. erikss. & ryvarden) jülich was described from decaying leaves only. d o m a ń s k i (1988, 1991, 1992) reported 20 saprobic species on angiosperms leaves and 7 species on fallen needles. this group includes trechispora gillesii (maas geest.) liberta and tubulicium capitatum (d.p. robers & boquiren) burds. & nakasone described from dead leaves only, the lattest species from palms, and leptosporomyces galzinii (bourdot) jülich sometimes inhabiting fallen leaves submerged in stream water. the list of species collected by us on fallen leaves of angiosperms includes amphinema byssoides, athelia arachnoidea (berk.) jülich, a. epiphylla complex, botryobasidium laeve (j. erikss.) parmasto, hyphodontia floccosa (bourdot & galzin) j. erikss., leptosporomyces galzinii, phanerochaete sanguinea, ph. sordida, steccherinum fimbriatum, tylospora fibrillosa. besides, several fungi were observed on fallen leaves adhered to decaying wood debris – botryobasidium subcoronatum (höhn. & litsch.) donk, chondrostereum purpureum, cylindrobasidium evolvens, peniophora cinerea (fig. 4) or to old polypore pileus – sistotrema brinkmannii (bres.) j. erikss. (fig. 5). the species collected by us on fallen coniferous needles are amphinema byssoides, athelia epiphylla complex, phanerochaete sanguinea, piloderma fallax, sistotrema brinkmannii, tomentella fuscocinerea (pers.) donk. several corticioid species are important inhabitants of plant rhizosphere in boreal forests, forming ectomycorrhiza, especially piloderma byssinum (p. karst.) jülich and p. fallax. thanatephorus pennatus currah and ypsilonidium sterigmaticum (bourdot) donk (in state of rhizoctonia) were isolated from mycorrhizal roots of orchids (d o m a ń s k i 1992). ceratobasidium cornigerum in state of rhizoctonia goodyerae-repentis auct. was isolated as endophyte from orchid roots (wa r c u p , ta l b o t 1966). we observed a complex of amphynema byssoides and tomentella vegetative hyphae associated with vaccinium myrtillus roots (fig. 6). d o m a ń s k i (1988, 1991, 1992) classifies 10 species in corticiaceae s. l. as litterinhabiting or colonizing different debris on soil. some of plant debris decay fungi 118 e. o. yurchenko have the ability to envelope by their basidiomata any meeting substratum, e.g. corticium boreoroseum boidin & lanquetin. the same author (1988, 1991, 1992) reported 18 species known from herbaceous stems and herb remains, including leaves. e.g. acanthobasidium delicatum (wakef.) oberw., epithele typhae (pers.) pat., and hypochnicium detriticum were documented for cyperaceae leaves. there are several species described in literature, specialized to inhabit some herbs, e.g. on cyperaceae (acanthobasidium delicatum), on monocotyledoneae (epithele typhae, phlebiella aurora (berk. & broome) k.h. larss. & hjortstam), on poaceae (acanthobasidium phragmitis), on saccharum (phanerochaete sacchari (burt.) burds.), on typha (hyphoderma typhicola (burt.) donk; j ü l i c h 1984; d o m a ń s k i 1988, 1991). tomentella cladii wakef. is reported by j ü l i c h (1984) on cladium mariscus (l.) pohl. only and tomentella juncicola svrček on juncus only. the species known on equisetum are athelopsis lembospora (bourdot) oberw., hypochnicium detriticum, and sistotrema pteriphilum k.h. larss. & hjortstam (d o m a ń s k i 1988, 1992). we collected fungi on dead stems of chamaenerion angustifolium (ceratobasidium aff. pseudocornigerum m.p. christ., peniophora cinerea, sistotrema octosporum (j. schröt.) hallenb.) and humulus lupulus l. (aleurodiscus cf. cerussatus (bres.) höhn. & litsch., msk 4963). chamaenerion angustifolium was mentioned as host for tomentella coerulea (kõljalg, 1996). besides, dead herbaceous stems attached to dead wood and bark were observed to be covered by fruitbodies of peniophora cinerea (msk 5287), and phanerochaete sordida (msk 6619). the colonization of dead grass blades arranged closely to wood was recorded for botryobasidium candicans j. erikss. (msk 4433), hyphoderma mutatum (msk 4098), h. setigerum (msk 4431b), peniophora cinerea (msk 4573, 5287), and phanerochaete sanguinea (4459). there are several species of ceratobasidium and thanatephorus occurring on living herbs and belonging to economically meaningful crop pathogens, though their parasitic activity is sometimes controversial. among them there are ceratobasidium oryzae-sativae p.s. gunnell & r.k. webster and c. setariae (sawada) p.s. gunnell & r.k. webster on rice and other crops (g u n n e l l , we b s t e r 1987); thanatephorus corchorus c.c. tu et al. causing damping-off of corchorus capsularis l. (tu et al. 1977); thanatephorus cucumeris (a.b. frank) donk – the most studied and known from over 200 hosts, mostly herbaceous ones, occurring mostly on stem bases and roots (d a n i e l s 1963); th. praticola (kotila) flentje growing on vegetables (d e s i l v a & wo o d 1964). besides, athelia rolfsii (curzi) c.c. tu & kimbr. in state of sclerotium rolfsii sacc. causes leaf and stem blights and fruit damage of many hosts (p u n j a et al. 1982; tu et al. 1992). other herb parasites are limonomyces roseipellis stalpers & loer. and l. culmigenus (j. webster & d.a. reid) stalpers & loer. on poaceae and cyperaceae (d o m a ń s k i 1991) and thanatephorus langlei-regis d.a. reid described from plantago lanceolata l. (r e i d 1969). thanatephorus orchidicola warcup & p.h.b. talbot is known from living orchids (wa r c u p , ta l b o t 1966), but also was found on living fern (k o t i r a n t a , s a a r e n o k s a 1993). herbaceous pteridiophyta sometimes draw special attention as substratum for corticiaceae s. l., e.g. h j o r t s t a m & l a r s s o n (1997) list 79 species on ferns. handbook by j ü l i c h (1984) reports 16 saprobic species on ferns, among them mycostigma aegeritoides (bourdot & galzin) jülich, parvobasidium cretatum (bourdot & gal natural substrata for corticioid fungi 119 zin) jülich, pteridomyces galzinii (bres.) jülich, and repetobasidiellum fusisporum j. erikss. & hjortstam known only from fern fronds, especially rhachises. h j o r t s t a m et al. (1988: 1471) indicate phlebiella filicina (bourdot) k.h. larss. & hjortstam as obligate fern inhabitant. d o m a ń s k i (1988, 1991, 1992) reported 26 species on herbaceous fern organs and fern debris, among them pteridomyces bananisporus boidin & gilles and p. capitatus boidin & gilles described from ferns only, and athelia pyriformis (m.p. christ.) jülich known on living ferns. tubulicium vermiculare was found on both herbaceous and arboreous ferns. two specimens from this ecological group were collected in belarus on dead fronds – phanerochaete sordida (msk 6619) and sistotrema sp. (msk 5564). bryophyta the substratum associations of fungi with true mosses and hepatics are rather rich, but not enough documented by corticiologists. the fungi found on living mosses can be classified into the two groups: embracing moss sprouts close to dead wood or bark, where the basidioma grows (fig. 7, 8) and occupying sprouts without clear connection of basidiomata with wood (fig. 9). evidently for a number of fungi moss sprouts serve for enlarging spore-producing surface and uplifting them above the ground. but in process of fungus-moss interaction a part of living moss organs become deformed, agglutinated and chlorophyll-less, indicating the evident negative fungus effect (fig. 7). a list of 19 species found on mosses can be extracted from j ü l i c h ’ s handbook (1984), from which tomentella brevispina is reported on moss only. ceratobasidium bicorne j. erikss. & ryvarden was published as known only from living polytrichum (e r i k s s o n , r y v a r d e n 1973). d o m a ń s k i (1988, 1991) reported 24 species on mosses including lindtneria leucobryophila (henn.) jülich, described on this substratum only, and lichenized fungi athelia phycophila jülich (known from moss only), dictyonema irpicinum mont., d. moorei (nyl.) henssen, d. pavonia (sw.) parmasto, d. sericeum (sw.) berk. bryophyta come into interaction with fungi from the earliest ontogenetic stages, e.g. we observed the association of living embryo states of unknown moss with tomentella sublilacina (ellis & holw.) wakef. hyphae on picea abies bark (msk 6570). in a previous paper (yu r c h e n k o 2001) we described the associations of 35 corticioid fungi with 11 moss species. the most common union with bryophyta forms amphinema byssoides, which hyphal strands, basidiomata patches and individual hyphae occur on living and dead lower parts of ground mosses hylocomium, pleurozium, and ptilium. the same habitat is frequently occupied by athelia epiphylla complex. frequent moss sprouts colonization is observed for fungi with actively growing hyphal strands and rhizomorphs, e.g. phanerochaete spp. and steccherinum fimbriatum. the biggest number of fungal species was collected in association with the genus brachythecium. lophocolea heterophylla (schrad.) dum. is a wood-inhabiting hepatic which living thalli most frequently overgrown by corticioid fungi, e.g. tomentella fuscocinerea, tubulicrinis subulatus (bourdot & galzin) donk (fig. 7), and tylospora fibrillosa. 120 e. o. yurchenko algae the substratum relations with algae is very poorly known phenomenon. the most prominent example is athelia arachnoidea, causing lesions in algal films covering bark of trees and bushes, and described in details by us (yu r c h e n k o , g o l u b k o v 2003). evidently other fungi are able to colonize epiphytic algae cover, but without such destructive activity, e.g. hyphodontia rimosissima (peck) gilb. which fruitbody was found growing in some areas on well discernible layer of green coccoid algae (fig. 10). lichenized fungi the inhabitation of lichens by corticiaceae s. l. is also poorly documented. the species mentioned on living and dead lichens in main handbooks are amphinema byssoides, athelia epiphylla s. l. (athelia epiphylla pers., a. salicum pers.), and sistotrema muscicola (pers.) s. lundell (e r i k s s o n , r y v a r d e n 1973; j ü l i c h 1984; d o m a ń s k i 1988, 1992). the active parasitizing of living lichens is known for us for athelia arachnoidea only. r y p a č e k (1967) discussed the antibiotic action of lichens on wood decay fungi in natural environment. he concluded that lichen metabolites, mostly lichen acids, supress mycelial growth and enzymatic activity of fungi in wood. nevertheless, we observed a number of resupinate non-poroid homobasidiomycetes coming into dense interaction with lichen thalli, e.g. botryobasidium candicans, cylindrobasidium evolvens, peniophora cinerea (fig. 11), p. nuda (fig. 12), phlebiella sulphurea, schizopora paradoxa (schrad.) donk, sistotrema brinkmannii, sistotremastrum suecicum. non-lichenized fungi and myxomycetes corticiaceae s. l. were observed on various taxonomic groups of other fungi, including ascomycetes, polyporoid and agaricoid homobasidiomycetes, and conidiomata of anamorphic fungi. pyrenomycete stromata serve as substratum e.g. for hyphoderma setigerum (on daldinia concentrica (bolton) ces. & de not., msk 6210; also yu r c h e n k o , z m i t r o v i c h 2001) and peniophora cinerea (on diatrypella favacea (fr.) ces. & de not., msk 6430). xenasma aculeatum c.e. gómez was reported on hypoxylon ascomata (d o m a ń s k i 1992). sometimes corticioid basidiomata occupy apothecia and subiculum of discomycetes, e.g. tapesia (fig. 13). corticium quercicola was reported to be regularly associated with old colpoma ascomata (j ü l i c h 1984). the inhabiting of decaying basidiomata of one fungus by another growing fungus is rather well known phenomenon for wood-inhabiting aphyllophorales in general. the highest ability to colonize other aphyllophoroid fungi we observed for sistotrema brinkmannii: it occurred on fomitopsis pinicola (sw.) p. karst. (msk 6419), ganoderma lucidum (curtis) p. karst. (msk 6212), peniophora incarnata (msk 4543), p. quercina (msk 6296), resupinate phellinus sp. (msk 5804), and stereum hirsutum (msk 5771). sometimes substratum relations between two corticioid fungi can be rather complicated, e.g. the fruitboidies grow on each other in different areas. example shown on figure 14 is hymenochaete tabacina occupying peniophora nuda basidioma, but in the center of photo a patch of p. nuda margin going on h. tabacina is visible. natural substrata for corticioid fungi 121 two fungi are known as evidently specialized parasites on other corticiaceae s. l.: galzinia forcipata pouzar on elaphocephala and laetisaria arvalis burds. hyperparasitic on thanatephorus cucumeris (d o m a ń s k i 1988, 1991). polypores are frequently occupied by resupinate non-poroid homobasidiomycetes, which was reflected in many publications, e.g. 33 species were listed by b e s l et al. (1989). j ü l i c h (1984) reported 7 species growing on old polypore fruitbodies. large polypores not rarely are used by corticioid fungi as main nutritive substratum. for instance, a fruitbody of phanerochaete laevis (pers.) j. erikss. & ryvarden totally overgrowing fomes fomentarius (l.) j.j. kickx hymenophore is shown on fig. 15. associations with agarics is more rare event. marasmius androsaceus (l.) fr. rhizomorphs and pileus-less stipes is a common component of coniferous forest litter in belarus, and e.g. hyphodontia breviseta (p. karst.) j. erikss. (msk 6222) was recorded on this substratum. colonization of armillaria spp. rhizomorphs attached to decaying wood was observed for amphinema byssoides (msk 4473) and hyphoderma praetermissum (p. karst.) j. erikss. & å. strid (fig. 16). anamorphic fungi fruitbodies as substratum for corticioid fungi are still neglected by mycologists. an example is peniophora cinerea several times observed on growing or decaying exosporium tiliae link conidiomata on dead tilia cordata branches (yu r c h e n k o 2001). peniophora incarnata was also recorded once on exosporium (fig. 17; coll. ye. rotkina). pycnidia of mitosporic fungi occurring on decaying wood are rather frequently enveloped by corticioid basidiomata. among our collections are sistotremastrum niveocremeum (höhn. & litsch.) j. erikss. covering spore-producing and destroying pycnidia of chaetodiplodia sp. (msk 4716) and tubulicrinis subulatus on diplodia-like fungus (fig. 18). very rarely corticioid fungi occur on myxomycetes. among our collections is peniophora incarnata (msk 4618) totally overgrowing a group of myxomycete sporocarps on wood. soil and humus the collective of corticioid species inhabiting soil is rather numerous, but they in general bulk are neglected in lists reviewing local mycobiotas. commonly such species also inhabit litter. their fructifications occur especially in soil interstices, e.g. mammal burrows. some species belong to the fungi regularly occurring in upper soil layer, e.g. piloderma fallax and tylospora fibrillosa. the observations of piloderma fallax in belarus permit to classify it more strictly as lower litter horizon fungus. the list of fungi growing on ground and humus extracted by us from j ü l i c h ’ s handbook (1984) includes 28 species, from which 3 species belong to the genus byssocorticium, 10 species to tomentella, and 4 species to tomentellastrum; the species tomentella fragilis (bourdot & galzin) m.j. larsen, t. nitellina bourdot & galzin, tomentellastrum fuscocinereum (pers.) svrček, t. litschaueri (svrček) m.j. larsen are reported only on soil and humus. d o m a ń s k i (1988, 1991, 1992) listed on soil and humus 27 species, including 4 species of sistotrema, 4 species of trechispora, lichenized athelia andina jülich, dictyonema pavonia, and d. sericeum, and the fungi reported only on soil or in soil: conohypha terricola (burt) jülich, echinotrema clanculare park.-rhodes, sistotrema hypogaeum warcup & p.h.b. talbot, waitea circinata warcup & p.h.b. talbot. the remarkable observations of peniophora lauta h.s. jacks. and tomentella 122 e. o. yurchenko fusca (pers.) schröt. fructifications on soil clods in cultivated fields were made by wa r c u p and ta l b o t (1963). occasionally typical wood fungi occur on soil particles close to decaying wood, as peniophora incarnata (msk 4682). mineral substrata several species were reported from stone surface in natural circumstances: athelia andina, piloderma lapillicola jülich, scopuloides hydnoides (cooke & massee) hjortstam & ryvarden, tomentella calcicola (bourdot & galzin) m.j. larsen, t. subcinerascens litsch., tomentellastrum caesiocinereum svrček (j ü l i c h 1984; d o m a ń s k i 1992). tomentella radiosa (p. karst.) rick was reported on sand (k õ l j a l g 1996). we collected tomentella bryophila (pers.) m.j. larsen underside of limestone piece in crimean forest (msk 5985). house fungi are known to be colonizing artificial stony materials, e.g. coniophora marmorata desm. (j ü l i c h 1984) and serpula lacrymans (wulfen) j. schröt. (msk 12127) were observed on concrete. growth of s. lacrymans is known on different calcium-containing mineral materials (b e c h a n d e r s e n 2005), bricks, oven clay, and even on glazed tile (b o n d a r t s e v 1948). leucogyrophana olivascens (berk. & m.a. curtis) ginns & weresub (msk 4945) was collected on mixed substratum of sand, subclay, and brick pieces in cellar. other substrata there are several exotic substrata not mentioned above. apples at store are colonized by athelia rolfsii (p u n j a 1982) and butlerelfia eustacei weresub & illman (we r e s u b , i l l m a n 1980) causing their decay. rare cases are associations of fruitbodies with chitine invertebrate debris, like hyphoderma setigerum (msk 4431b), in which thickened up to 7 mm basidioma tightly embraces, incorporates in and grows through big chitine exoskeletons of an unidentified insect. the using of living nematodes as nutritive substratum is a scarcely investigated fact, known for probable hyphoderma sp. with stephanocysts (l i o u , t z e a n 1992). single species was described from dung – dacryobasidium coprophilum (wakef.) jülich, but it can also colonize leaves and twigs (j ü l i c h 1984). two species of trechispora, t. polygonospora ryvarden and t. spinulifera jülich were found on termitaries (d o m a ń s k i 1992). hypochnicium eichleri (bres.) j. erikss. & ryvarden was collected once on earth-worm excrements (e r i k s s o n , r y v a r d e n 1976), which can be indeed classified as a type of humus substratum. should be mention that serpula lacrymans can grow on different artificial seminatural (fibre board, gypsum plates, wallpaper, textiles) and synthetic (polyurethane foam) materials (b e c h -a n d e r s e n 2005). we observed this fungus on old book (ex cwu sine no.). conclusion the data accumulated in literature and by us show that the range of substrata for corticioid fungi in situ includes all types of terrestrial plant bodies, both living and dead, other fungi, both lichenized and non-lichenized, humus, mineral bodies, and in exclusive cases invertebrates and shallow water submerged plant debris. our ob natural substrata for corticioid fungi 123 servations demonstrated that typically wood-inhabiting fungi can be found on different non-woody substrata. in most cases these are the particles attached or adjacent to decaying wood or bark. some litter fungi, especially those with rhizomorphs of hyphal cords, have the ability to cover any meeting substratum. frequently fungi colonize living mosses near wood or bark surface with outer observing negative effect on them. lichens and algae are much less documented substrata for corticiaceae s. l. than other cryptogams (mosses and ferns). colonization of different kinds of artificial mineral, semi-natural, and synthetic materials is still known for house fungi only, especially for serpula lacrymans. acknowledgements: the work was partly supported by grant b05 171 of belarusian republican fund for fundamental research. i am grateful to hbl. dr g.f. rykovsky (v.f. kuprevich institute of experimen tal botany, minsk) for identifications of some bryophyta and to dr d.i. tret’yakov (the same institution) for identifications of some vascular hosts. i am thankful to mr. a.yu. akulov (mycology and plant resist ance department, kharkiv national university, cwu myc herbarium) for giving a duplicate of serpula lacrymans growing on book. the materials of this paper were partly presented as poster at xiv congress of european mycologists. i met prof. alina skirgiełło first there and she become interested in my research into interaction of corticioid fungi with other organisms. thanking in that number to this meeting the materials become summarized and promulgated. references a a n d s t a d s., r y v a r d e n l. 1987. aphyllophorales on wooden fences in norway. windahlia 17: 49 54. b e c h a n d e r s e n j. 2005. the true dry rot fungus, serpula lacrymans. history, occurrence in nature and distribution in europe. (in:) v.g. s t o r o z h e n k o , v. i . k r u t o v (eds). problems of forest phyto pathology and mycology. proceedings of the 6th international conference. moscow, petrozavodsk: forest research institute of karelian research centre ras. p. 34 43. b e s l h., h e l f e r w., l u s c h k a n. 1989. basidiomyceten auf alter porlingsfruchtkörpern. ber. bayer. bot. ges. 60: 133 145. b i n d e r m., h i b b e t t d.s., l a r s s o n k. h., l a r s s o n e., l a n g e r e., l a n g e r g. 2005. the phy logenetic distribution of resupinate forms across the major clades of mushroom forming fungi (ho mobasidiomycetes). systematics and biodiversity 3 (2): 113 157. b o n d a r t s e v a.s. 1948. on the distribution of house fungi in leningrad over last years (1940 1946). priroda 11: 37 42. d a n i e l s j. 1963. saprophytic and parasitic activities of some isolates of corticium solani. trans. brit. mycol. soc. 46 (4): 485 502. d o m a ń s k i s. 1988. mała flora grzybów. basidiomycetes (podstawczaki). aphyllophorales (bezblasz kowce). 5. corticiaceae: acanthobasidium irpicodon. pwn, warszawa kraków, 427 pp. d o m a ń s k i s. 1991. mała flora grzybów. i. basidiomycetes (podstawczaki). aphyllophorales (bezblasz kowce). stephanosporales (stefanosporowce). 6. corticiaceae: kavinia rogersella, stephanospora ceae: lindtneria. pwn, warszawa kraków, 272 pp. domański s. 1992. mała flora grzybów. i. basidiomycetes (podstawczaki). aphyllophorales (bezblasz kowce). 7. corticiaceae: sarcodontia ypsilonidium, christiansenia and syzygospora. w. szafer insti tute of botany, polish academy of sciences, kraków, 258 pp. e r i k s s o n j., r y v a r d e n l. 1973. the corticiaceae of north europe. vol. 2: aleurodiscus conferto basidium. oslo: fungiflora. p. 60 286. e r i k s s o n j., r y v a r d e n l. 1976. the corticiaceae of north europe. vol. 4: hyphodermella mycoa cia. oslo: fungiflora. p. 547 886. g u n n e l l p.s., we b s t e r r.k. 1987. ceratobasidium oryzae sativae sp. nov., the teleomorph of rhizoc tonia oryzae sativae and ceratobasidium setariae comb. nov., the probable teleomorph of rhizoctonia fumigata comb. nov. mycologia 79 (5): 731 736. h a w k s w o r t h d.l., k i r k p.m., s u t t o n b.c., p e g l e r d.n. (eds) 1995. ainsworth and bisby’s dic tionary of the fungi. 8th ed. wallingford: cab international. 616 pp. 124 e. o. yurchenko h j o r t s t a m k., l a r s s o n k. h. 1997. corticioid fungi growing on ferns in northern europe. windahlia 22: 49 55. h j o r t s t a m k., l a r s s o n k. h., r y v a r d e n l. 1988. the corticiaceae of north europe. vol. 8: phle biella, thanatephorus ypsilonidium. oslo: fungiflora. p. 1450 1631. j ü l i c h w. 1984. die nichtblätterpilze, gallertpilze und bauchpilze. aphyllophorales, heterobasidio mycetes, gastromycetes. (in:) h. g a m s (ed.) kleine kryptogamenflora. band iib/1. basidiomyce ten. 1. teil. stuttgart, n.y.: g. fischer. 626 pp. k i r k p.m., c a n n o n p.f., d a v i d j.c., s t a l p e r s j.a. (eds) 2001. ainsworth and bisby’s dictionary of the fungi. 9th ed. egham, wallingford: cab international. 655 pp. k õ l j a l g u. 1996. tomentella (basidiomycota) and related genera in temperate eurasia (synopsis fun gorum. vol. 9). oslo: fungiflora, 1996. 213 pp. k o t i r a n t a h., s a a r e n o k s a r. 1993. rare finnish aphyllophorales (basidiomycetes) plus two new combinations in efibula. ann. bot. fennici 30: 211 249. l i o u j.y., t z e a n s.s. 1992. stephanocysts as nematode trapping and infecting propagules. mycologia 84 (5): 786 790. n a k a s o n e k.k., g i l b e r t s o n r.l. 1978. cultural and other studies of fungi that decay ocotillo in arizona. mycologia 70 (2): 266 299. p a r m a s t o e., nilsson h., larsson k. h. 2004. cortbase version 2. extensive updates of a nomencla tural database for corticioid fungi (hymenomycetes). phyloinformatics 1: 5. p u n j a z.k., g r o g a n r.g., a d a m s g.c. 1982. influence of nutrition, environment, and the isolate on basidiocarp formation, development, and structure in athelia (sclerotium) rolfsii. mycologia 74 (6): 917 926. r e i d d.a. 1969. new or interesting british plant diseases. trans. brit. mycol. soc. 52 (1): 19 38. r y p a č e k v. 1967. biologie dřevokazných hub [wood decay fungi biology. a translation of czech edi tion, expanded and supplemented by the author. translated by m. gashkova, edited by a.t. vanin]. moscow: lesnaya promyshlennost’. 276 pp. d e s i l v a r.l., w o o d r.k.s. 1964. infection of plants by corticium solani and c. praticola effect of plant exudates. trans. brit. mycol. soc. 47 (1): 15 24. tu c.c., c h e n g y.h., k i m b r o u g h j.w. a new species of thanatephorus from jute in taiwan. myco logia 69 (2): 409 413. tu c.c., h s i e h t.f., ts a i w.h., k i m b r o u g h j.w. 1992. induction of basidia and morphological comparison among isolates of athelia (sclerotium) rolfsii. mycologia 84 (5): 695 704. wa r c u p j.h., talbot p.h.b. 1963. ecology and identity of mycelia isolated from soil. ii. trans. brit. mycol. soc. 46 (4): 465 472. wa r c u p j.h., ta l b o t p.h.b. 1966. perfect states of some rhizoctonias. trans. brit. mycol. soc. 49 (3): 427 435. we r e s u b l.k., i l l m a n w.i. 1980. corticium centrifugum reisolated from fisheye rot of stored apples. can. j. bot. 58: 137 146. y u r c h e n k o e.o. corticioid fungi on mosses in belarus. mycena (1) 1: 71 91. y u r c h e n k o e.o., g o l u b k o v v.v. 2003. the morphology, biology, and geography of a necrotrophic basidiomycete athelia arachnoidea in belarus. mycological progress 2 (4): 275 284. y u r c h e n k o e.o., z m i t r o v i c h i.v. 2001. variability of hyphoderma setigerum (corticiaceae s. l., ba sidiomycetes) in belarus and northwest russia. mycotaxon 78: 423 434. naturalne substraty dla grzybów korticjoidalnych s t r e s z c z e n i e autor podaje przegląd typów substratów zasiedlanych przez grzyby nieporoidalne przytacza jąc przykłady z literatury i własnych obserwacji na białorusi. jako podłoża dla grzybów wyróżnia drzewa, krzewy i zielne rośliny naczyniowe, mszaki, glony, grzyby zlichenizowane i niezlicheni zowane, a także śluzowce i bezkręgowce. grzyby występują na żywych i obumarłych, w różnych stadiach rozkładu organizmach oraz ich częściach. ponadto grzyby znane są jako występujące na glebie, próchnicy, skałach, sztucznych, syntetycznych materiałach oraz na odchodach. 2014-01-01t11:43:50+0100 polish botanical society hebelomina neerlandica, a new species for ukraine and considerations about the genus hebelomina andré fraiture1 and vera hayova2 1jardin botanique national de belgique domaine de bouchout, b 1860 meise, fraiture@br.fgov.be 2department of mycology, m. g. kholodny institute of botany, national academy of sciences of ukraine tereshchenkivska 2, 01601 kiev, veha@ln.ua f r a i t u r e a., h a y o v a v.: hebelomina neerlandica, a new species for ukraine and considerations about the genus hebelomina. acta. mycol. 41 (2): 177 188, 2006. hebelomina neerlandica huijsman has been found near kiev (ukraine). it is an addition to the ukrainian mycoflora. the specimens are described and illustrated. a synopsis of the genus hebelomina is presented, with comments on the systematics and the distribution of its different species, which are all very rare. key words: hebelomina, hebeloma, gymnopilus, rapacea, cortinariaceae, ukraine introduction during a mycological excursion in a forest near novobilychi (kiev, ukraine), a whitish gilled mushroom has been observed. it was growing on a dead log of pinus sylvestris lying on the ground. several sporophores have been collected. their characters are typical of hebelomina neerlandica huijsman, a.o. the general habit, whitish colour, ecology, size and shape of the cheilocystidia, size and shape of the spores, which are smooth, whitish and dextrinoid. this species has never been reported from ukraine. a short description of the specimens is given below. up to now, six species have been described in the genus hebelomina, most of them are extremely rare or even known only by the type specimens. the taxonomic position of the genus and its species is still under discussion. the nomenclature is also quite complicated. a summary of the data concerning those questions is presented here as well as a list of the descriptions and illustrations published for the different species, an overview of their worldwide distribution and an identification key. acta mycologica vol. 41 (2): 177-188 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 178 a. fraiture and v. hayova description of the ukrainian collection sporophores completely whitish, drying ochraceous, sometimes with a rather dark rusty brown spot. pileus 12-22 mm broad, convex with involute margin, smooth, moist, not viscid. lamellae adnate. stipe 12-25 x 2-4.5 mm, central, cylindrical, curved, without annulus, small remnants of a cortina-like velum sometimes visible on young specimens. context whitish. no particular smell detected. taste not tested. spores (figs 1a and 2) (6.0-) 6.5-7.5 (-8.0) x (3.5-) 4.0-4.5 (-5.1) μm, generally amygdaliform, sometimes ellipsoid or ovoid, hyaline, smooth, without germ-pore, slightly thick-walled, dextrinoid (reddish-brown in melzer reagent). basidia (fig. 1b) irregularly cylindrical, (2-) 4-spored, 28-30 x 5-7 μm, with sterigmata 4-6 μm long. cheilocystidia (fig. 1c) usually narrowly lecythiform, sometimes narrowly lageniform, narrowly conical or cylindrical, 25-35 x 5.0-6.5 μm (body), x 2.0-3.0 μm (neck) and x 2.5-4.5 μm (head). pleurocystidia absent. clamp-connections present. ecology. sporophores not caespitose, growing on a fallen dead trunk of pinus sylvestris, in a forest of pinus sylvestris with numerous quercus robur, on podsolic sand. specimen described. ukraine, kiev, forest near novobilychi, 14. ix. 2004. herbarium a. fraiture 2927 (br), double in herb. v. hayova (kw 29993). fig. 1. hebelomina neerlandica: a spores, b basidia, c cheilocystidia (specimen a. fraiture 2927, br); scale bar = 10 μm. hebelomina neerlandica, a new species for ukraine 179 synopsis of the genus hebelomina genus hebelomina maire 1935, bull. soc. hist. nat. afrique du nord 26: 13. ≡hebeloma [sect. denudata] subsect. hebelomina (maire) beker, eberhardt and vesterholt 2005, in vesterholt, the genus hebeloma: 24 [note: this is probably the correct name]. type species: hebelomina domardiana maire. the taxonomic position of the genus hebelomina is not easy to circumscribe and the question has been treated by several authors. the most argumented discussion is probably the one proposed by s i n g e r (1986: 611), who finally decides to accommodate the genus in the cortinariaceae, where he assumes it is related to cortinarius and leucocortinarius or, more probably, to hebeloma. the difficulty in positioning the genus in the systematics is partly due to its very specific characters, a.o. the spores which are whitish, smooth, thick walled, devoid of a pore and dextrinoid. as it will be seen hereunder, the problem also lies in the fact that the genus hebelomina is heterogenous. h. domardiana, type species of the genus hebelomina, seems to be a white spored hebeloma. consequently, the genus hebelomina has been recently included in hebeloma (ve s t e r h o l t 2005). however, it will not be possible to transfer to hebeloma all the species described in hebelomina. molecular analysis (m o n c a l v o et al. 2002: 367, 379) has shown that hebelomina neerlandica is probably a gymnopilus. those authors extend however improperly their conclusions to the whole genus hebelomina. ? hebelomina amazonensis sing. 1979, in singer and araujo, acta amazonica 9 (1): 32 [invalid: nomen nudum]. this is a simple mention of the name, without any description or citation of a specimen. s i n g e r did not cite the name in his agaricales in modern taxonomy, ed. 4 (1986) and one may thus suppose that he did not believe anymore in the value of this taxon. hebelomina domardiana maire 1935, bull. soc. hist. nat. afrique du nord 26: 13. ≡hebeloma domardianum (maire) beker, eberhardt and vesterholt 2005, in vesterholt, the genus hebeloma: 102 [note: this is probably the correct name of the species]. descriptions: m a i r e (1935, original description), ve s t e r h o l t (2005), u r b o n a s (2005: 174). illustrations: m a i r e (1935, line drawings of the specimens and microscopic characters), ve s t e r h o l t (2005, aquarelle of the specimen and line drawings of spores and cheilocysti dia), u r b o n a s (2005, pl. 21,1: a f, aquarelle of the specimens and line drawings of micro scopic characters). m a i r e (1935), in the first lines of his paper, already says that h. domardiana is «un champignon très remarquable, ayant l’aspect extérieur d’un tricholoma, mais qui est, en réalité, un hebeloma à spores incolores». the great mycologist also points out that the genus hebelomina «est aux hebeloma ce que le genre cortinellus [ leucocortinarius] est aux cortinarius». he adds in his comments that the amygdaliform spores, with an epispore rigid and thin but looking double, becoming violaceouspurple by iodine when young, the edge of the gills entirely covered with filamentous subclaviform and very dense hairs and even the faint raddish smell and the bitterness of the flesh are characters of hebeloma. it is possible to add other characters corresponding with the genus hebeloma: big spores (11-15 x 8 μm) with a clearly papil180 a. fraiture and v. hayova late top, white cap becoming reddish brown in the centre, white stem very pruinose under the gills (“valde pruinosa”, in italics in the latin diagnosis), tricholomoid habit and terricolous ecology. g e n n a r i (2003) says “su legno di quercus suber”, but the original description gives no indication concerning lignicolous ecology. moreover, maire compares his species with hebeloma and tricholoma and tells that, at first sight, he took it for hygrophorus eburneus var. pseudodiscoideus; all of those fungi are terricolous. molecular analysis recently confirmed the intuition of maire and showed that, despite its white spores, hebelomina domardiana probably belongs to the genus hebeloma, where it occupies however a “very isolated position” (ve s t e r h o l t 2005). unfortunately, it seems that the type specimen, collected by maire, has not been preserved or is lost. therefore, vesterholt had to undertake molecular study of material collected in estonia. the species has also been reported from lithuania and latvia (u r b o n a s , k a l a m e e s , l u k i n 1986). if there is no problem to admit that the different collections from the three baltic states belong to the same taxon, it is less easy to believe that a species growing in the baltic states can also be present in a quercus suber wood in algeria, especially for a taxon which is most probably mycorrhizal. if the type material can not be found again, it would be interesting to make a new collection of the species in its original growth place or in a quercus suber wood of the mediterranean region. distribution: algeria: forêt de l’alma, 15. xii. 1933, under quercus suber (maire 1935). maire says “mauritaniae” but l’alma is situated about 40 km to the east of alger; its name is now boudouaou (p. bertea and j.f. trimbach, comm. pers.). no herbarium specimen is formally cited in the protologue. after h o r a k (1968: 266) and h u i j s m a n (1978: 487), the type specimens have been lost. estonia: surju, 28. viii. 1989, in mixed forest, specimen ve s t e r h o l t jv89 497 (ve s t e r h o l t 2005: 133). latvia: on humus in pine forests; no locality and no collecting date indicated (u r b o n a s et al. 1986: 71). g e n n a r i (2003) men tions a personal communication from kalamees, after which the collections have been lost. after n e z d o i m i n o g o (1996), those records are quite doubtful. lithuania (u r b o n a s 2005: 174): rūdininkų, šalčininkų distr., 03.viii.1974, in mixed forest with pinus, young quercus and betula. viešvilés reserve, jurbarko distr., 19.vii.2000, young picea abies forest. biržų forest, biržų distr., 16.viii.2001, in mixed forest. see also u r b o n a s et al. (1986: 71, cf. comments given here above for latvia) hebelomina maderaspatana natarajan and raman 1980, kavaka 8: 72. descriptions and illustrations: n a t a r a j a n and r a m a n (1980, original description and line drawings of the specimens, spores and basidia). the text and the illustrations, except the latin description, are reproduced in n a t a r a j a n and r a m a n (1983: 137 138 et fig. 22, e g). the classification of this species is very unclear. the limoniform and rather big spores (7-11.2 x 5.6-8.4 μm) are quite typical of hebeloma while the presence of rhizomorphs at the base of the stem indicates rather a saprotrophic species growing on litter or in connection with wood. on the other hand, the reddish-brown to orange-brown colours are rather resembling the genus gymnopilus. moreover, the lack of cheilocystidia corresponds neither with hebeloma nor with gymnopilus and therefore h. maderaspatana could belong to another genus. distribution: india: campus of the indian institute of technology, guindy (madras), 23. viii. 1978, on ground, in groups, coll. n. raman (mubl 2421, paratypus) ibid., 3.xi.1978, on lit ter, in groups, coll. n. raman (mubl 2420, holotypus) (n a t a r a j a n , r a m a n 1980, 1983). hebelomina neerlandica, a new species for ukraine 181 hebelomina mediterranea a. gennari “2002”, publ. 2003, riv. micol. 45 (4): 312. descriptions and illustrations: g e n n a r i (2003, original description and colour photo graph of the specimens, spores, cheilocystidia and epicutis; line drawings of the spores, ba sidia, cheilocystidia and epicutis). the big limoniform to subamygdaliform spores, measuring 9-11 (-12) x 6-8 (-8.5) μm, the subclavate to cylindrical cheilocystidia, the milky white cap, with a cream ochraceous centre, the stem pruinose on the apex, the tricholomatoid habit and the terricolous ecology are connecting the species with the genus hebeloma. it is very probable that its pertaining to this genus will be demonstrated. the species appears to be very close to h. domardiana. the comparison of the descriptions (g e n n a r i 2003; m a i r e 1935; u r b o n a s 2005; ve s t e r h o l t 2005) shows only a few differences between the two species. the specimen of h. mediterranea is more robust than h. domardiana (cap diameter 5-7 cm versus 2.5-4 cm) and its spores are slightly shorter: 9-11 (-12) μm instead of 11-15 or 10.7-13.4 μm. gennari also mentions a germ-pore (“poro germinativo evidente”) and represents it on his drawings. however, true germ-pores have not been described in hebeloma and the structures seen by gennari correspond probably with a callus rather than a pore (s i n g e r 1986; p e g l e r , yo u n g 1971; m e l é n d e z -h o w e l l 1967). the description of gennari is based on a single collection and can not provide any idea of the variability of the species. in conclusion, we consider as possible that h. mediterranea could be conspecific with h. domardiana. distribution: italy: civitella in val di chiana, prov. arezzo, 15.x.2002, terricolous, col lected among a mediterranean vegetation composed of quercus ilex, q. pubescens, arbutus unedo, erica scoparia, cistus monspeliensis and c. salvifolius, leg. silvia urci (mcve 669, ho lotypus) (g e n n a r i 2003). hebelomina microspora alessio and nonis 1977, micol. ital. 6 (3): 19; non h. microspora huijsman [ h. neerlandica]. descriptions and illustrations: a l e s s i o , n o n i s (1977, original description, aquarelles and photographs of the specimens). note: a l e s s i o (1981) mentions that the colours of the plate have not been accurately reproduced, that the aquarelles are actually of a very pale cream ivory ochre with which contrast much more coloured patches, reaching the «terra cot ta» colour; he also says the red colour of the photographs is too marked and diffuse. the affinities of this taxon are difficult to circumscribe. the fusoid to cylindrical cheilocystidia, the top of the stem slightly white pruinose and the creamy white cap, becoming pale ochre and finally slightly brownish with irregular pale clay-orange patches are characters of hebeloma. but, on the other hand, the ovoid to amygdaloid spores, measuring 6-8 (-9) x 4-4.5 μm, and the lignicolous ecology are rather in favour of the genus gymnopilus. hebelomina microspora “huijsman ex” alessio and nonis 1977 is probably another species than h. microspora huijsman [ h. neerlandica, see that species]. distribution: italy: parco della rimembranza augustae taurinorum, torino, prov. pie monte, 5.v.1976, on slightly emergent root of pinus strobus, alt. 500 600 m, leg. bruna nonis (herb. e. rebaudengo, cebae, holotypus) (a l e s s i o , n o n i s 1977). the authors indicate «vere autumnoque», what should signify that another observation was done in autumn. punta manara, sestri levante, prov. liguria, 27.xii.1989, at the base of a pinus pinaster trunk, leg. r. m. dameri (o r s i n o , tr a v e r s o 1990). 182 a. fraiture and v. hayova hebelomina neerlandica huijsman 1978, persoonia 9 (4): 490 [nom. nov. to replace h. microspora huijsman 1978, non h. microspora alessio and nonis 1977]. ≡hebelomina microspora huijsman 1946, rev. mycol. ns 11: 31 [invalid, art. 36.1]; hebe lomina microspora huijsman “ex huijsman” 1978, persoonia 9 (4): 485 [illeg., art. 53.1] non h. microspora alessio and nonis (1977). ≡hebelomina huijsmaniana singer 1986, agaricales in modern taxonomy (ed. 4): 612 [illeg., art. 52.1; nom. nov. to replace h. microspora huijsman 1978, non h. microspora alessio and nonis 1977]. descriptions: g a r n w e i d n e r (1996), h u i j s m a n (1946, 1978, original description), n e v i l l e , r o u x (1997), s p o o n e r (1993), vo l d e r s (1997), u r b o n a s (2005). illustrations: a n o n y m o u s (2003, colour photograph of the specimens), g a r n w e i d n e r (1996, colour photograph of the specimens and line drawings of spores and cheilocystidia), h u i j s m a n (1946, line drawings of microscopic characters of the typus; 1978, line drawings of the type specimens), n e v i l l e and r o u x (1997, colour photograph and line drawings of microscopic characters), s p o o n e r (1993, line drawings of spores and cheilocystidia), vo l d e r s (1997, line drawings of microscopic characters; the author also mentions a slide jvdm 7677 and an aquarelle o. van de kerckhove 295, br, both unpublished), u r b o n a s (2005, line drawings of microscopic characters). the publication of a valid name for this taxon has not been easy. the species is first described by h u i j s m a n (1946), under the name hebelomina microspora. unfortunately, no latin diagnosis is provided and the name is thus invalid (art. 36.1). about thirty years later, a l e s s i o and n o n i s (1977) publish a latin diagnosis in order to validate the name created by huijsman. however, they choose another type than the specimen cited by huijsman and, consequently, at a nomenclatural point of view, they create a new taxon. on the other hand, as pointed out by different authors (a.o. h u i j s m a n 1978; n e v i l l e , r o u x 1997), there are several morphological differences between the two specimens : the italian collection has a inocyboid habit, reddish brown colours on the pictures, amyloid spores and twice bigger cheilocystidia. therefore, they probably belong to two different taxa at a systematic point of view as well. that hypothesis is not accepted by a l e s s i o (1981), who believes that a single taxon is involved. we consider that conspecificity possible, while unlikely. a few months after a l e s s i o and n o n i s (1977) and h u i j s m a n (1978) also decides to validate his species by publishing a latin diagnosis. his paper is at the point to be sent to the printer when he discovers the “validation” made by alessio and nonis. he publishes nevertheless his own “validation” (which is illegitimate: later homonym, art. 53.1) but, thinking that his fungus is not the same that the one of the italian authors, he adds, at the end of his text, a note in which he proposes hebelomina neerlandica as a new name for his species. would the synonymy be proven in the future, then the species should be named h. microspora alessio and nonis and h. neerlandica would be reduced to a superfluous synonym. finally, s i n g e r (1986), reading the h u i j s m a n (1978) paper, overlooks its final note and introduces hebelomina huijsmaniana as a new name to replace h. microspora huijsman. the name published by singer is illegitimate (superfluous name, art. 52.1). the nomenclatural changes are not finished yet. the species will probably be transferred to the genus gymnopilus, since molecular analyses have shown that the species is likely a white spored gymnopilus, rather close to g. penetrans (m o n c a l v o et al. 2002: 367, 379). regarding the macroand microscopic characters of hebelomina neerlandica, a new species for ukraine 183 the species as well as its ecology on pinus wood, it seems reasonable to consider h. microspora as a whitish gymnopilus, with white and smooth spores. within that genus, the absence of a membranous ring, the relatively small size of the spores and the growth on coniferous wood correspond with the group of g. stabilis, g. sapineus, g. penetrans and g. hybridus. the status of the three last species is still uncertain and varies considerably in the recent literature: three (m o s e r 1983; o r t o n 1993) or two independent species (h o l e c 2005) or one single species (h ø i l a n d 1990). distribution: belgium: bois des manants, tilff, prov. liège, 6.x.1978, 31.x.1978 and 5.x.1979, on dead wood of pinus sylvestris (branches of various size, stumps, cones), herb. v. demoulin (lg). “de kuik”, gooreind (wuustwezel), prov. antwerpen, 9.xi.1996, in a pine wood on sandy soil, on fallen branches and litter of pinus nigra, herb. o. van de ker ckhove 499 (br), j. volders 96205 and a. de haan 96100 (volders 1997). the species has been found every year at the same place up to the year 2000 (d e h a a n 2001). france: “croix de novy”, monregard, dépt. haute loire, 23.ix.1993, on a piece of rotten coniferous wood (picea abies, abies alba or pinus sylvestris), leg. a. c h a r r e t (herb. p. neville 93.09.20.25 and p. roux 93.09.17.57) (n e v i l l e , r o u x 1997). forêt de pont calleck, dépt. morbi han, x.2002, on rotten wood of pinus, two collections leg. p. hériveau and leg. r. chalange (herb. m. chiaffi) (a n o n y m o u s 2003 and g. e y s s a r t i e r , comm. pers.). germany: schöngeising, near fürstenfeldbruck, distr. oberbayern, 13.xi.1994 and 1.x.1995, on a dead larix decidua trunk, herb. e. garnweidner (g a r n w e i d n e r 1996). lithuania: žagarés forest, joniškio distr., 20.ix.1990, on fallen rotting branches striniškų forest, vilkaviškio distr., 25.vii.1999, on fallen rotting branches (u r b o n a s 2005: 174) netherlands: near rijssen, prov. overijssel, 24.x.1943, on dead twigs of pinus sylvestris, coll. w.j. reuve camp and w.f. smits (l, holotypus) (h u i j s m a n 1946, 1978). the growth place has been de stroyed soon after 1945, when a new quarter of rijssel was built; moreover, the typus has been found in a very poor condition, badly moth eaten and mouldy (h u i j s m a n 1978). «de fonteintjes», south of rijssen, 1988, under picea abies, coll. c. bas, and 1990, leg. w. ligterink (l) (unpublished data, cited fide vo l d e r s 1997; a r n o l d s , k u y p e r and n o o r d e l o o s 1995; see also the distribution map in a n o n y m o u s 2000). ukraine: forest near novo bilychi (kiev), kiev obl., 14.ix.2004, on dead log of pinus sylvestris lying on the ground, herb. a. fraiture 2927 (br) and v. hayova (kw 29993) (this publication). united kingdom: surrey, oxshott heath, nr bog n of sandy lane, 14.x.1984, under pinus and betula on damp sandy ground, coll. l. spooner (k) ibid., 13.x.1991, on dead twig in litter, herb. kew (k) (s p o o n e r 1993). south hampshire, 1999, clustered on fallen branch of salix (bmsfrd n°480383). north hampshire, 1999, on mossy fallen branch of salix in broadleaf semi natu ral woodland (bmsfrd n° 493526) (british mycological society fungus record database). hebelomina pallida dessi and contu 1993, in contu and dessi, micol. veget. medit. 8 (2): 104. descriptions and illustrations: c o n t u , d e s s i (1993, original description, with colour photograph [the legend of which erroneously mentions (hebelomina candida) and habit sketch of the carpophores and line drawings of the spores, basidia and cheilocystidia). the authors of the species point out that h. pallida is close to h. neerlandica and they list the following characters to separate the two species: the carpophores of h. pallida have a white colour remaining nearly unchanged during their whole life, they are completely devoid of a veil even in very young stage, they have bigger and non amygdaliform spores and they grow on dead wood of eucalyptus. those differences are not much significative. h. neerlandica is also a whitish species and the colour modifications described by huijsman (becoming pale ochraceous-aluta184 a. fraiture and v. hayova ceous, often more or less mixed with incarnate) may considerably vary depending on ecological conditions. the veil of that species is fugacious and often difficult if not impossible to see. as for the spore size, the figures are indeed a bit bigger: 7.5-9.0 (-10.3) x 5.2-6.0 (-6.9) μm for h. pallida versus 6.5-7.8 x 4.2-4.6 μm for h. neerlandica (h u i j s m a n 1978), but some collections of the latter species have shown bigger spores (a.o. vo l d e r s 1997). the difference between the two species is mainly significative for the spore width. since the estimation of the spore length/width ratio (q) for h. pallida is about 1.47 when calculated on the spore size reported by c o n t u , d e s s i (1993) and about 1.77 when calculated after the drawings provided by the same authors, it seems possible that the spore size given by dessi and contu is not perfectly accurate. the habitat on eucalyptus wood is indeed unusual but not sufficient to create a new species. in conclusion, we think that h. pallida is very close to h. neerlandica and even possibly conspecific with that species. distribution: italy: serramanna, prov. cagliari, sardinia, 05.xii.1992, on dead wood of eucalyptus camaldulensis, herb. m. contu 92/269 (cag, holotypus) ibid., 06.xii.1992 and 10.i.1993, leg. p. dessi and m. contu (cag and pers. herb. p. dessi, paratypus) (c o n t u , d e s s i 1993). identification key to the species described in hebelomina 1) cheilocystidia absent. carpophores reddish brown to orange brown. india. ............................................................................................................................. h. maderaspatana 1) cheilocystidia abundant. carpophores usually much paler, often whitish. europe, algeria ............................................................................................................................................. 2 2) terricolous species. spores bigger than 9 x 6 μm, often more or less citriform or with a pap illate top. cheilocystidia mostly cylindrical or clavate, sometimes narrowly lageniform .................................................................................................................... (hebelomoid species) 3 2) lignicolous species. spores smaller than 9 x 6 μm, usually amygdaliform or ellipsoid and not citriform. cheilocystidia mostly narrowly lecythiform, except in h. microspora, where they are fusoid to cylindrical. ....................................................................... (gymnopiloid species) 4 3) cap 5 7 cm broad. spores 9 11 ( 12) μm long. italy. .................................... h. mediterranea 3) cap 2.5 4 cm broad. spores 11 15 μm long. algeria and baltic countries. ................................................................................................................................... h. domardiana 4) spores amyloid. cheilocystidia bigger than 40 x 9 μm, mostly fusoid to cylindrical. on pine wood. may and december. italy ............................................................................ h. microspora 4) spores dextrinoid. cheilocystidia smaller than 40 x 9 μm, mostly narrowly lecythiform. on coniferous wood or on eucalyptus. september january. europe ............................................ 5 5) spores (3.5 ) 4.0 5.0 ( 5.2) μm wide. carpophores whitish, becoming pale ochraceous al utaceous or incarnate. on coniferous wood, rarely on salix. (september ) october novem ber. europe. ............................................................................................................. h. neerlandica 5) spores 5.2 6.0 ( 6.9) μm wide. carpophores whitish and remaining so. on eucalyptus wood. december january. italy ................................................................................................ h. pallida hebelomina neerlandica, a new species for ukraine 185 summary and conclusions the genus hebelomina is heterogenous. it can be divided into at least two groups of species. 1) the hebelomoid species (h. domardiana and h. mediterranea). they are terricolous and probably ectomycorrhizal, with a tricholomoid or hebelomoid habit, a stem which is pruinose in its upper part, rather big spores (usually above 9 x 6 μm), more or less citriform or amygdaliform with a papillate top, cheilocystidia usually irregularly cylindrical or clavate, eventually narrowly lageniform but not lecythiform. both species have been described from mediterranean oak forests but, rather unexpectedly, the first one has also been reported from the three baltic countries, in pine forests and in mixed forests. the two species are very close to the genus hebeloma. this has been recently confirmed by molecular analysis for h. domardiana (specimen from estonia), which has consequently been transferred to that genus (ve s t e r h o l t 2005). we suggest that the two species are close to each other and even possibly conspecific. on the other hand, the baltic collections attributed to h. domardiana could belong to a separate, undescribed species of this group. 2) the gymnopiloid species (h. neerlandica, h. pallida and probably h. microspora). they are lignicolous and saprotrophic, with a gymnopiloid habit, a stem not or only slightly pruinose, medium sized to small spores (usually under 9 x 6 μm), which are amygdaliform, ellipsoid or ovoid and neither citriform nor papillate; the cheilocystidia are usually narrowly lecythiform. most of the collected specimens of h. neerlandica were growing on coniferous wood, mainly pinus sylvestris, but also p. nigra, larix decidua and picea abies; there are two reports on salix in great britain. h. microspora has been observed on wood of pinus strobus and p. pinaster. h. pallida is only known from the type collections, on wood of eucalyptus camaldulensis. by its morphological characters, h. neerlandica is very close to gymnopilus. this has been confirmed by molecular analysis (m o n c a l v o et al. 2002) and the species will probably be transferred to that genus. h. pallida is very close to h. neerlandica and could even be conspecific. we believe that h. microspora belongs to this same group; however, its taxonomic position is less easy to interpret. 3) incertae sedis (h. maderaspatana). this species exhibits characters from the two groups cited above and, besides, the lack of cheilocystidia does not fit with the genera hebelomina, hebeloma and gymnopilus. the taxonomic position of the species is thus still unclear. in conclusion, it seems probable that most of the species of the genus hebelomina will be transferred to either hebeloma or gymnopilus and that the genus hebelomina will disappear. it is nevertheless noteworthy that most of the species described in hebelomina share some original characteristics which separate them from those two genera. the carpophores are often whitish or very pale, at least when young. the spores are very particular, being whitish and smooth under the light microscope when the spores in hebeloma and gymnopilus are brown and rather coarsely ornamented. it seems that it is difficult to obtain a good spore print because the spores, while being usually produced in large amount, are remaining on the gills. when it was possible to obtain a spore deposit from a hebelomina collection, it has been observed that it was not pure white but very pale brownish. the pictures of the spores of h. neerlandica, seen by sem (fig. 2) show a kind of shallow ornamentation although it could be an 186 a. fraiture and v. hayova artefact due to insufficient reinflation. the presence of those special features in the different hebelomina species could be explained by a mutation, inducing the loss of pigmentation of the carpophores and changing the brown and ornamented spores of hebeloma and gymnopilus into whitish and smooth “hebelominoid” spores. an unpublished paper by g a s p a r i n i (pers. comm.) has drawn our attention to the genus rapacea, created by h o r a k (1999) to accommodate a single species, rapacea mariae e. horak. it is a cortinarioid species, recorded from new zealand, tasmania and papua new guinea, which seems to be another example of this “hebelominoid syndrome”, again affecting a species of the cortinariaceae family. it has whitish carpophores (see colour picture in s o o p 2005) and its spores are pale olivaceous-argillaceous, i.e. much paler than the normal colour of the spores in the genus cortinarius. they are inamyloid, but become brown in melzer’s reagent ( dextrinoid), and appear smooth under the light microscope but minutely asperulate or with low net-like ridges under sem. molecular and phylogenetic analyses have shown that rapacea was nested in the genus cortinarius (p e i n t n e r et al. 2002a) and, consequently, the species has been transferred to that genus: cortinarius mariae (e. horak) e. h o r a k et al., in p e i n t n e r et al. (2002b: 449). acknowledgements. the authors thank the mycologists who sent them information through the internet forum mycologia europaea, particularly g. eyssartier, for the unpublished de scription of a collection, and p. bertea and j. m. trendel, for the localization of the forêt de l’alma. they also thank h. beker for the interesting discussion concerning the topic, v. demoulin (lg) for providing copies of three articles and giving access to his specimens, g. adamonyte and j. kasparavicius (bilas) for sending copy of a reference and b. gasparini for the communication of an unpublished paper. they are grateful to o. van de kerckhove (br) for preparing the drawings for publication, m. verhaegen (br) for taking the pictures with the electronic microscope and d. aplin (br) for helping to solve some questions about english language. references a l e s s i o c.l., n o n i s u. 1977. una specie quasi sconosciuta: hebelomina microspora. micol. ital. 6 (3): 15 19 + pl. 18. a l e s s i o c.l. 1981. revisione di miei lavori comparsi nei primi 25 numeri di micologia italiana. micol. ital. 10 (1): 21 27. a n o n y m o u s 2000. kaartenbijlage overzicht van de paddestoelen in nederland, 2 vols. nederlandse mycologische vereniging, baarn. 349 + 329 pp. a n o n y m o u s “2003”, 2004. session de la société mycologique de france à guidel (morbihan) du 21 au 26 octobre 2002. bull. trim. soc. mycol. fr. 119 (3/4): 385 402. a r n o l d s e., k u y p e r th.w., n o o r d e l o o s m.e. 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(in:) r. wa t l i n g , n.m. g r e g o r y (eds). british fungus flora, agarics and boleti, 7 cortinariaceae p.p.: 58 72. royal botanic garden edinburgh. p e g l e r d.n., yo u n g t.w.k. 1971. basidiospore morphology in the agaricales. beih. nova hedw. 35: vi, 210 pp. + 53 pl. p e i n t n e r u., h o r a k e., m o s e r m., v i l g a l y s r. 2002a. phylogeny of rozites, cuphocybe and rapa cea inferred from its and lsu rdna sequences. mycologia 94 (4): 620 629. p e i n t n e r u., h o r a k e., m o s e r m., v i l g a l y s r. 2002b. rozites, cuphocybe and rapacea are taxo nomic synonyms of cortinarius: new combinations and new names. mycotaxon 83: 447 451. s i n g e r r., a r a u j o i.j.s. 1979. litter decomposition and ectomycorrhiza in amazonian forests, 1 a comparison of litter decomposing and ectomycorrhizal basidiomycetes in latosol terra firme rain forest and white podzol campinarana. acta amazonica 9 (1): 25 41. s i n g e r r. 1986. the agaricales in modern taxonomy (4th ed.). koeltz scientific books, koenigstein, viii, 981 p. + 88 pl. s o o p k. 2005. cortinarioid fungi of new zealand. an iconography and key (4th ed.). scientrix, mora, 100 pp. s p o o n e r b.m. 1993. hebelomina neerlandica huijsman in persoonia 9: 490 (1978). mycologist 7 (3): 108. u r b o n a s v. 2005. nuosedieciai (cortinariales). lietuvos grybai 8 (5): 288 pp. + 32 pl. u r b o n a s v., k a l a m e e s k., l u k i n v. 1986. conspectus florum agaricalium fungorum (agaricales s.l.) lithuaniae, latviae et estoniae [ed. 2]. mokslas, vilnius. 140 pp. ve s t e r h o l t j. 2005. the genus hebeloma. fungi of northern europe 3: 1 146. svampetryk, tilst, danemark. vo l d e r s j. 1997. hebelomina neerlandica huijsman, de witspoorvaalhoed, een nieuwe soort en genus voor belgië. amk meded. 97 (1): 2 5. 188 a. fraiture and v. hayova hebelomina neerlandica, gatunek nowy dla ukrainy i rozważania o rodzaju hebelomina s t r e s z c z e n i e owocniki hebelomina neerlandica huijsman zostały niedawno zebrane w okolicach kijo wa. ten nowy dla ukrainy gatunek został opisany i zilustrowany przez autorów, którzy rów nocześnie dyskutują pozycję taksonomiczną rodzaju, a w nim sześciu dotychczas znanych ga tunków. są to: h. maderaspatana, h. mediterranea, h. domardiana, h. microspora, h. pallida, h. neerlandica. praca zawiera klucz do oznaczania, analizę literatury, na podstawie której rozważana jest delimitacja taksonów, nazewnictwo, cechy taksonomiczne i rozmieszczenie. w większości są to gatunki rzadko spotykane, niektóre znane tylko z pojedynczych okazów. dokładne wyjaśnienie wielu kwestii poruszanych przez autorów wymaga dalszych zbio rów, do których zachętą może być niniejsza praca. 2014-01-01t11:44:10+0100 polish botanical society gloiodon strigosus (swartz: fr.) p. karst. (bondarzewiaceae) in poland anna bujakiewicz department of plant ecology and environment protection, adam mickiewicz university umultowska 89, pl-61-614 poznań, ascom@amu.edu.pl b u j a k i e w i c z a.: gloiodon strigosus (swartz: fr.) p. karst. (bondarzewiaceae) in poland. acta mycol. 4� �1�:69-��, ����.4� �1�:69-��, ����. gloiodon strigosus �swartz: fr.� p. karst. recognized as an extinct species in poland, has been recently found in the białowieża national park. iconography and synonyms are given and the distribution and ecology is discussed. key words: gloiodon strigosus, bondarzewiaceae, russulales, extinct species introduction gloiodon strigosus �swartz: fr.� p. karst., a representative of bondarzewiaceae family �k i r k et al. ���1�, recognized as an extinct species in the red list of macrofungi in poland �wo j e w o d a , ł a w r y n o w i c z ���6� has been recently found in the area of strict protection of the białowieża national park. fructifications were collected by the author in september ���4 during the mycological excursion devoted to the observation of fruit bodies of a rare and interesting fungus, rhodotus palmatus which occurs in that area regularly in early autumn, since ���1 �b u j a k i e w i c z ����b, ����; b u j a k i e w i c z , n i t a ���4�. the locality site is characteristic of dense thickets covering piles of fallen logs of elm (ulmus scabra� which fell a victim of the dutch elm disease. the logs are not removed and nourish many rare representatives of plants, animals �insects� and fungi �b u j a k i e w i c z ����a, b, �����. nomenclature, iconography and drawings hydnum strigosum swartz, kongl. vetensk. acad. nva handl. �1���: �5�, 181�: fr., syst. mycol. 1: 414, 18�1; gloiodon strigosus �swartz: fr.� p. karst., medd. soc. f. fl. fenn. 5: 4�. 18�9; sclerodon strigosus �swartz: fr.� karst., finl. basids. �61. 1889. n i k o l a j e v a �1961� ��5-���, fig. 15�-155, j a h n �19�9� �5, fig. 4�; j a h n and s t u r m �198��, fig. 1-1�; r y m a n and h o l m å s e n �1984�: 1�8; k o s k i -k o t i r a n t a and n i e m e l ä 1988 �198��: 61-64, fig. 1�-15. acta mycologica vol. 4� �1�: 69-�� ���� dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday �� a. bujakiewicz description of collected material fructifications of gloiodon strigosus were collected on september 18, ���4 in forest section �98 of the area of strict protection of the białowieża national park �fig. 1� along the “didactic trial”, on underneath of the log of ulmus scabra, on fig. 1. the białowieża forest �acc. to fa l i ń s k i 1986, modified� – distribution of gloiodon strigosus in the białowieża forest. 1-locality in the area of strict protection of the białowieża national park ����4�. gloiodon strigosus �1 decorticated wood in association with auricularia mesenterica. the fruit body was emerging from remnants of an older one and was mostly fan shaped �fig. ��. the forest is classified as the phytocoenosis of the fraxino-alnetum association with the elements of the tilio-carpinetum association. with regards to morphology and sporulation elements of the fruit bodies specimens collected correspond entirely with j a h n ’ s �19�9� photo �fig. �� and j a h n and s t u r m ’ s �198�� description. minutely verrucose spores of gloiodon strigosus seen under the electron microscope are presented on figure �. there are three species up to now recognized in the genus gloiodon: g. occidentale ginns growing on gymnosperms in north america and having glabrous spores, g. nigrescens �petch� maas occurring in sri lanka, having pileal surface with scattered hairs or even glabrous and g. strigosus distinguished by the occurrence on angiosperm wood, having pileal surface densly haired and smaller verrucose spores. �g i n n s 1988�. gloiodon strigosus resembles auriscalpium vulgare in many characters, both macro -and microscopical and formerly belonged to the auriscalpiaceae family (m a a s g e e s t e r a n u s 196��. now it belongs to bondarzewiaceae family and represents the order russulales �k i r k et al. ���1; wo j e w o d a �����. in the literature gloiodon strigosus is recorded on angiosperms, mostly on populus, alnus, salix and prunus, seldom is found on ulmus and betula (k o s k i -k o t i r a n t a , n i e m e l ä 1988�. n i k o l a j e v a �1961� noted some siberian fructifications collected in sajan mts on fir �abies� which may belonged to g. occidentale. gloiodon strigosus is saprotrophic, causes white rot �j a h n 19�9� and prefers humid microclimate. it is a rare and endangered species with vulnerable ecology, connected with old, well preserved forests. it’s distribution covers the northern hemisphere mainly throughout the boreal zone, both in the interior of the continents and in oceanic areas. the first find of gloiodon strigosus in central europe was announced by j a h n and s t u r m �198�� in the bavarial alps in an oroboreal montane zone �k o s k i -k o t i r a n t a , n i e m e l ä 1988�. distribution in poland: ladzka forest in the complex of the białowieża primeval forest, on logs of deciduous trees �b ł o ń s k i 1889�; area of strict protection of the białowieża national park, forest section �98. in the world: scandinavia: finland, norway, sweden �r y v a r d e n 19�1; s t r i d 19�5; i n g e l ö g et al. 1984�, estonia �j ä r v a , p a r m a s t o 198��, czech republic, france, hungary �j ü l i c h 1984�, ukraina �z e r o v a et al. 18���, siberia �n i k o l a j e v a 1961�, russia far east �l y u b a r s k i j , va s i l i e v a 19�5�, india �j a h n , s t u r m 198�� and north america in both canada �p o m e r l e a u 198�� and the usa �b a n k e r 191�; h a r r i s o n 19��� �k o s k i -k o t i r a n t a , n i e m e l ä 1988�. aknowledgements. i express my thanks and appreciation to mr marek snowarski �wrocław� for photo taken to fresh fructifications of gloiodon strigosus and to the laboratory of electron and confocal microscopy of the adam mickiewicz university in poznań for examing fungus and professional service. �� a. bujakiewicz references b a n k e r h. 191�. type studies in the hydnaceae 6. the genera creolophus, echinodontium, gloiodon and hydnodon. mycologia 5: �9�–�98. b ł o ń s k i f. 1889. spis ro�lin zarodnikowych zebranych lub zanotowanych w lecie w r. 1888 w puszf. 1889. spis ro�lin zarodnikowych zebranych lub zanotowanych w lecie w r. 1888 w pusz-puszczach: białowieskiej, świsłockiej i ladzkiej �list of cryptogamic plants collected or noted in summer of 1888 in primeval forests: puszcza białowieska, świsłocka and ladzka�. �in:� f. b ł o ń s k i , k. d r y m m e r �eds�. sprawozdanie z wycieczki botanicznej odbytej do puszczy białowieskiej, ladzkiej i świsłockiej w 1888 roku �report of the excursion to primeval forests: puszcza białowieska, ladzka and świsłocka in 1888�. pam. fizj. 9: 55–115 �in polish�. b u j a k i e w i c z a. ����a. new, rare and endangered fungi in the białowieża primeval forest �e poland�. polish bot. j. 4� ���: 11�–1�4. b u j a k i e w i c z a. ����b. rhodotus palmatus �bull.: fr.� r. maire. �in:� w. w o j e w o d a �ed.�. atlas of the geographical distribution of fungi in poland. �. w. szafer institute of botany, polish academy of sciences, kraków: 95–98. b u j a k i e w i c z a. ����. puszcza białowieska ostoją rzadkich i zagrożonych grzybów wielkoowoc-ch grzybów wielkoowocnikowych. parki nar. rez. przyr. �� ���: ���–�46. b u j a k i e w i c z a., n i t a j. ���4. żyłkowiec różowawy rhodotus palmatus �bull.: fr.� r. maire – mieszkaniec białowieskich ostępów. chrońmy przyr. ojcz. 6� �5�: 8�–85. f a l i ń s k i j. b. 1986. vegetation dynamics in temperate lowland primeval forest. ecological studies in białowieża forest. �in:� m. j. a. we r g e r �ed.�. geobotany 8, dr w. junk publishers. dordrecht. g i n n s j. 1988. new genera and species of lignicolous aphyllophorales. mycologia 8� �1�: 6�–�1. h a r r i s o n k. 19��. aphyllophorales �. hydnaceae and echinodontiaceae. �in:� g. a i n s w o r t h , f. s p a r r o w , a . s u s s m a n �eds�. the fungi, an advanced treatise 4b. a taxonomic review with keys: basidiomycetes and lower fungi. new york–london: �69–�95. i n g e l ö g t., t h o r g., g u s t a f s s o n l. 1984. floravård i skogobruket �. uddevalla, 4�8 pp. j a h n h. 19�9. pilze die an holz wachsen. herford. j a h n h., s t u r m ch. 198�. der seltene stachelpilz gloiodon strigosus �sw. ex fr.� p. karst. in den alpen gefunden. westfäl. pilzbr. 1�-11: ��9–���. j ä r v a l., p a r m a s t o e. 198�. eesti seente kondimestik. tartu, ��1 pp. j ü l i c h w. 1984. die nichtblätterpilze, gallertpilze und bauchpilze �aphyllophorales, heterobasidiomycetes, gastromycetes� �in:� h. g a m s �ed.�. kleine kryptogamenflora. ii b/1. basidiomyceten 1. fisher verl., stuttgart–new york, 6�6 pp. k i r k m. p., d a v i d p. f., s t a l p e r s j. c. ���1. ainsworth & bisby’s dictionary of the fungi. 9 th ed. cab international, wallinford, 655 pp. k o s k i -k o t i r a n t a s., n i e m e l ä t. 1988. hydnaceous fungi of the hericiaceae, auriscalpiaceae and climacodontaceae in northwestern europe. karstenia ��: 4�–��. l y u b a r s k i j l., va s i l y e v a l. 19�5. drevorazrušayuščie griby dalnego vostoka. novosibirsk,164 pp. m a a s g e e s t e r a n u s r.a. 196�. hyphal structures in hydnums ii. proc. kon. nederl. akad. wetensch. �ser. c� 66: 4�6–45�. n i k o l a j e v a t. 1961. ežovikovye griby. �in:� v. s a v i č �ed.�. flora sporovych rastenij sssr 6 �griby ��. moskva & leningrad, 44� pp. p o m e r l e a u r. 198�. flore des champions an quebéc et regions limitrophes. montréal, 65� pp. r y m a n s., h o l m å s e n i. 1984. svampar, en fälthandbok. stockholm, �18 pp. r y v a r d e n l. 19�1. studies in the aphyllophorales of firnmark, northern norway. rep. kevo. subarctic res. sta. 8: 148–154. s t r i d å. 19�5. wood – inhabiting fungi of alder forests in north-central scandinavia. 1. aphyllophorales (basidiomycetes�. taxonomy, ecology and distribution. wahlenbergia 1: 1–���. w o j e w o d a w. ����. checklist of polish larger basidiomycetes. �in:� z. m i r e k �ed.�. biodiversity of poland. �. w. szafer institute od botany, polish academy of sciences, kraków, 81� pp. w o j e w o d a w., ł a w r y n o w i c z m. ���6. red list of the macrofungi in poland. czerwona lista grzy-czerwona lista grzybów wielkoowocnikowych w polsce. �in:� z. m i r e k , k. z a r z y c k i , w. w o j e w o d a , z. s z e l ą g �eds�. red list of plants and fungi in poland. czerwona lista ro�lin i grzybów polski. w. szafer insti-red list of plants and fungi in poland. czerwona lista ro�lin i grzybów polski. w. szafer institute of botany, polish academy of sciences, kraków: 5�–��. z e r o v a m., r a d z i e v s k i j g., š e r č e n k o s. 19��. viznačnik gribiv ukrajni. 5. basidiomiceti 1, ekzobasidialni, afilofaralni, kantarelalni. kijv, �4� pp. gloiodon strigosus �� gloiodon strigosus �swartz: fr.� p. karst. �bondarzewiaceae) w polsce s t r e s z c z e n i e przedstawiono opis stanowiska, synonimikę i ikonografię gloiodon strigosus, gatunku uważanego za wymarły w polsce. podano też uwagi dotyczące ekologii i chorologii gatunku. gloiodon strigosus jest gatunkiem wskaźnikowym dobrze zachowanych borealnych lasów łęgowych. fig. 2. fructifications of gloiodon strigosus: a) hymenophore with spines; b) upper surface covered with soft hairs. phot. m. snowarski. a b fig. 3. minutely verrucose spores of gloiodon strigosus (sem x 6,78 kx) (courtesy of lab. electron & confocal microscopy, adam mickiewicz university, poznań). 2014-01-01t11:45:14+0100 polish botanical society 2014-08-25t21:39:04+0200 polish botanical society cristulariella depraedans as causal agent of leaf spots of a maple and other trees and shrubs tadeusz kowalski and czesław bartnik department of forest pathology, university of agriculture in kraków al. 29. listopada 46, pl-31-425 kraków, rltkowal@cyf-kr.edu.pl kowalski t., bartnik cz.: cristulariella depraedans as causal agent of leaf spots of maple and other trees and shrubs. acta mycol. 43 (1): 5–12, 2008. results of the first studies in poland on cristulariella depraedans, the causal agent of necroses on tree and shrub leaves, are presented. the fungus was recorded on acer platanoides, a. pseudoplatanus and eight other species of trees and shrubs, including corylus avellana, fagus sylvatica, lonicera xylosteum and padus avium that were not previously known as hosts of the fungus. the disease symptoms, which depend on the host plant species, the size and number of necrotic lesions on leaves, and the morphological features of the fungal propagules were characterized. key words: cristulariella depraedans, acer spp., leaf spots, host range introduction many and various symptoms with abiotic and biotic causes occur on leaves of maple trees, acer platanoides l. and a. pseudoplatanus l. only some of them are easily recognizable.these include symptoms caused by fungi, e.g. rhytisma acerinum (pers.) fr. and r. punctatum (pers.) fr. (butin 1996; mańka 2005) or by gall-forming insects and mites, e.g. harrisomyia vitrina (kieffer) and eriophyes pseudoplatani corti (skrzypczyńska 2004). the joint action of fungi and gall-forming insects in the development of necroses can make the diagnosis of their causal agents difficult. kowalski (2003) recorded 18 species of fungi within the necrotic areas accompanying the galls caused by drisina glutinosa giard. the most significant were colletotrichum gloeosporioides (penz.) sacc., diplodina acerina (pass.) sutton, discula campestris (pass.) arx and phomopsis platanoidis died. similar associations were reported by wulf (1990), who found 12 species of fungi on leaves of a. pseudoplatanus within the necrotic tissues of galls caused by dasineura vitrina kieffer. a great variety of fungi can infect maple leaves and recognition of symptoms and signs caused by individual potential causal agents is necessary. acta mycologica vol. 43 (1): 5–12 2008 6 t. kowalski and cz. bartnik studies on diseases of forest trees in the south of poland led to the identification of cristulariella depraedans (cooke) hoehn., a fungus that occurred commonly on maple leaves in a few locations. since the species has not been recorded in poland previously, the objective of this study was to present an analysis of symptoms and signs of infection by the pathogen on maple tree leaves. the symptoms on leaves of the other species of trees and shrubs growing in proximity to the infected maples were also analysed to indicate the pathogen’s hosts range. materials and methods the study was carried out in 1996-2006, in four localities in the south of poland (tab. 1). the majority of observations were made in ojców national park, where the trees and shrubs along a route that included the łokietek cave, ojców centre, prądnik valley, the kraków gate and again the łokietek cave were surveyed and analysed. from july to october, 1-3 times a year, 30 to 60 leaves with different disease symptoms were collected from 30 trees, mostly self-sown, of acer pseudoplatanus and a. platanoides. samples were either from young trees or from the lower parts of the crowns of older trees. on each occasion, 3-6 diseased leaves from each of 20 other species of trees and shrubs growing in the surrounding area were also collected and assessed (tab. 2). their grouping according to the type of symptoms and the presence of sporulation of c. depraedans was made in the laboratory. the number and size of necrotic lesions caused by c. depraedans on 165 maple leaves collected at random from severely affected trees were evaluated. the sizes of similar lesions on leaves of the other species of trees and shrubs and the pigmentation of the necrotic tissues in the centres of the lesions and at their borders with the living leaf tissues were also determined. the propagules formed by c. depraedans were examined microscopically and a macroand microphotographic record was made of the symptoms and signs of infection by c. depraedans. the nomenclature of host plants is given after mirek et al. (2002). results cristulariella depraedans was recorded in four localities in the south of poland (tab. 1). it was common in ojców and tatra national parks and wolski forest (tab. 3) but occurred only sporadically in świerklaniec forest district. in ojców national park, most infection was recorded in 1996-1998, when more than 80% of trees growing along the route surveyed had local necrotic lesions caused by c. depraedans on their leaves. in a few young maple trees, up to 1 m high, all leaves in the crowns were infected (fig. 1). in tatra national park, świerklaniec forest district and wolski forest, c. depraedans occurred only on leaves of a. pseudoplatanus, and in ojców national forest it occurred on both species of acer and on eight species of other trees and shrubs, viz. carpinus betulus, cornus sanguinea, corylus avellana, fagus sylvatica, lonicera xylosteum, padus avium, quercus robur and tilia cordata. there were no symptoms of infection by c. depraedans on a further 12 species of trees and shrubs growing in the area surrounding the infected maples (tab. 2). cristulariella depraedans 7 some differences were observed in the symptoms caused by c. depraedans on a. pseudoplatanus and a. platanoides, and on other plant species. up to 438 necrotic lesions per leaf, with lesion diameters up to 22 mm, occurred on a. pseudoplatanus (tabs 2, 3). single lesions were almost round, greyish-white in the centre and dark grey at the edge (tab. 2; figs 1, 2). ta b l e 1 locations and periods of study and plant species examined (e, examined; n, no symptoms and not examined) area studied period of studies number of observations in a year species of plants studied acer platanoides acer pseudoplatanus other species of tress and shrubs ojców national park 1996-2002, 2006 2 3 e e e tatra national park 1999-2002, 2005 1 n e e świerklaniec forest district 1999-2002, 2006 1 2 n e e wolski forest 2001-2002 1 n e e list of species is given in table 2 ta b l e 2 presence, size and appearance of lesions caused by cristulariella depraedans on different trees and shrubs species of plant size of necrotic spots colour of centre colour of edge (mm) a. plants with cristulariella depraedans acer platanoides l. 1-33 light grey to dark grey to light brown dark brown acer pseudoplatanus l. 1-22 greyish-white dark grey carpinus betulus l. 1-3 grey to dark grey light brown brownish-grey cornus sanguinea l. 1-3 grey violet corylus avellana l. 1-3 light brown greyish-brown fagus sylvatica l. 1-2 grey grey or greyish-green lonicera xylosteum l. 1-5 whitish-grey dark grey padus avium mill. 1-3 whitish-grey greyish-brown or greyish-green sometimes with a purplish tint quercus robur l. 1-6 light beige light brown tilia cordata mill. 1-2 light grey dark grey b. plants with no cristulariella depraedans aesculus hipocastanum l. salix fragilis l. alnus glutinosa (l.) gaertn. sambucus nigra l. betula pendula roth. sorbus aucuparia l. fraxinus excelsior l. spiraea salicifolia l. populus tremula l. symphoricarpos albus (l.) s.f. blake quercus rubra l. ulmus glabra huds. 8 t. kowalski and cz. bartnik germinating c. depreadans propagules were often seen within the initial and smallest lesions. disintegration of the necrotic tissues caused irregular perforations between the leaf veins in a few of the more advanced lesions. single, adjacent lesions sometimes merged and the severely infected leaves rolled up, died and were shed prematurely, often as early as august. up to 338 local lesions per leaf occurred on a. platanoides (tab. 3). small lesions were 2–5 mm in diameter and larger lesions reached up to 33 mm in diameter (tab. 3, fig. 3). the lesions were light grey to light brown in the centre and dark grey to dark brown at the edge. as on leaves of a. pseudoplatanus, germinating propagules of c. depraedans were often seen within the initial and smallest lesions (fig. 4). lesions caused by c. depraedans on leaves of a. pseudoplatanus at ojców national park were larger than those recorded at the same time at tatra national park (tab. 3). ta b l e 3 numbers and size ranges of lesions caused by cristulariella depraedans on leaves of acer platanoides and a. pseudoplatanus at different periods of study studied area period of studies number of leaves summary number of necroses with diam (mm) (minimal and maximal number on a leaf) < 5 6-10 > 10 acer platanoides ojców national park viii.1997 10 706 63 73 (2-308) (2-16) (2-14) ojców national park viii.2000 10 710 146 30 (32-113) (8-26) (1-6) ojców national park viii.2001 10 340 52 46 (4-91) (0-11) (1-9) acer pseudoplatanus ojców national park viii.1997 30 4435 122 11 (12-373) (0-25) (0-6) ojców national park viii.2000 30 5863 423 12 (18-399) (1-37) (0-2) ojców national park viii.2001 30 4501 172 14 (42-321) (1-14) (0-3) tatra national park viii.2000 15 557 0 0 (9-107) (0-0) (0-0) tatra national park viii.2002 15 1721 46 2 (0-219) (0-11) (0-1) wolski forest x.2001 15 442 155 57 (3-104) (4 29) (0-8) cristulariella depraedans 9 leaves of other species of plants were infected by c. depraedans to different extents. the largest numbers of necrotic lesions per leaf were recorded on cornus sanguinea (up to 582) and on quercus robur (up to 286) (figs 7, 9). lesions diameters were between 1 and 6 mm (tab. 2). the necrotic tissues showed more or less distinctive differences in pigmentation of the central and peripheral parts in all the species of plants studied (tab. 2). the near-violet pigmentation of the lesion edges on leaves of cornus sanguinea and greyish-green pigmentation on leaves of padus avium and fagus sylvatica were the most characteristic (tab. 2, fig. 9). differences in pigmentation of these tissues were not species-specific. a similar range of pigmentation was observed during these studies on leaves of c. sanguinea infected by septoria cornicola desm., of p. avium infected by stigmina carpophila (lev.) m.b. ellis, and on a few leaves of f. sylvatica in the necrotic tissue surrounding galls caused by mikiola fagi (hartig) and inhabited by apiognomonia errabunda (rob.) hoehn. the whitish propagules of c. depraedans, which were clearly discernible against the greyish background of the lesions, were observed in the necrotic areas on the undersides of leaves and sometimes also on the upper surface of the leaf blades (fig. 5). propagules were bulb-shaped, round, slightly flatten or slightly elongated 60 – 140 µm in diameter, mounted on a septate, hyaline stipe, 120 – 180 µm long and 10 – 15 µm wide (fig. 6). the bulb, which consisted of many single, round cells, 6 – 9 µm in diameter, was readily separated from the stipe. disscusion the fungus cristulariella depraedans has been known since the end of 19th century, when it was described as polyactis depraedans cooke (cooke 1885). it is not a commonly occurring species. it has been recorded previously only in the united kingdom, germany, austria and the usa (cooke 1885; saccardo 1908; sydow 1912; bowen 1930; batko 1974; butin 1981a; cech, donaubauer 1990; wulf 1994). the present studies show that it also occurs in the south of poland, where it was first recorded in ojców national park. its occurrence in abundance was associated mainly with heavy rainfall in the growing season of 1996-1997 (feliksik et al. 2002; saramak 2005). the significance of wet conditions for the development of the fungus was also noticed and emphasized by bowen (1930), butin (1981b), cech and donaubauer (1990), wulf (1994) and lang (2000). favorable moisture conditions can be also found on the leaves of susceptible young trees regenerated naturally and growing under the canopy of older stands. in ojców and tatra national parks, favourable moisture conditions occur in the numerous depressions and valleys. for more than 100 years the host range of c. depraedans has been known to include various species of acer, bucida buceras l. and aruncus dioicus (walt.) fern. (redhead 1975; lang 2000). only recently studies in germany in 1996-1999 showed that c. depraedans has a much broader potential host range than known previously. lang (2000) reported its occurrence on 21 woody and herbaceous plants other than acer species. the present studies also confirmed that the host range of c. depraedans is not limited to plants within genus of acer. in ojców national park, the fungus was observed on eight other species of plants growing in the areas around the infected a. platanoides and a. pseudoplatanus. four of them, corylus avellana, fagus 10 t. kowalski and cz. bartnik sylvatica, lonicera xylosteum and padus avium, are new additions to the host range of c. depraedans. lang (2000) showed that the size and pigmentation of necrotic lesions and their edges, next to the living leaf tissues, depend on the species of host plant and are related to differences in their susceptibility. lesions were different in size on leaves of a. pseudoplatanus, a. platanoides and the other plant species studied. this undoubtedly resulted from the interval of time between infection and observation. it should be mentioned that propagules formed within lesions were transferred to other leaves and to other parts of the leaf that had the primary infection, and caused secondary infections at different times in the same growing season. occasionally, germinating fungal propagules were observed on the initial necroses, with diameters not exceeding 1 mm, in the second half of the growing season. despite the differences in lesion sizes resulting from the period of infection, effects of host plant on the lesion dimensions were also observed. the larger lesions on leaves of a. platanoides seem to suggest that it is more susceptible than a. pseudoplatanus. the propagules of c. depraedans found in poland did not differ morphologically from those found in other countries (redhead 1975; willetts 1997; butin 1981b; wulf 1994). their presence is essential for the identification of c. depraedans because the lesions are not specific and are very similar to those caused by other fungi or insects and mites causing galls (kowalski 2003; skrzypczyńska 2004). the present studies confirm that, on the plants examined, there was no co-occurrence of a related species, cristulariella moricola (hino) redhead (syn. c. pyramidalis waterman & marshall), which has propagules that are of different shape (redhead 1975; trolinger et al. 1978). acknowledgement. the study was supported by the ministry of science and higher education, grant no. 2 p06l 036 26. references batko s. 1974. notes on new and rare fungi on forest trees in britain. bull. br. mycol. soc. 8: 19–21. bowen p. r. 1930. a maple leaf disease caused by cristulariella depraedans. conn. agric. exp. stn. bull. 316: 625–647. butin h. 1981a. aktuelle baumkrankheiten im forst. mitt. biol. bundesanst. land – forstw. berlindahlem. 203: 177–178. butin h. 1981b. die weißfleckigkeit des bergahorns – eine „neue“ blattkrankheit. allg. forstzeitschrift. 14: 327-328. butin h. 1996. krankheiten der waldund parkbäume. thieme verlag. stuttgart. cech t. l., donaubauer e. 1990. cristulariella depraedans – weissfleckenkrankheit von ahornblättern. forstschutz aktuell. 12: 3. cooke m. c. 1885. some remarkable moulds. jour. quek. micr. club. s. ii, 2: 138–143. feliksik e., wilczyński s., durło g. 2002. charakterystyka zmienno�ci opadów atmosferycznych na kop-charakterystyka zmienno�ci opadów atmosferycznych na kopciowej koło krynicy zdroju w latach 1971-2000. acta agr. et silv., ser. silv. 40: 5–16. kowalski t. 2003. endophytic fungi: v. mycobiota in living leaves of acer pseudoplatanus and in necrotic tissues around galls of drisina glutinosa. phytopath. pol. 29: 23–35. lang k. j. 2000. new hosts of cristulariella depraedans. for. path. 30: 117–120. mańka k. 2005. fitopatologia le�na. pwril. warszawa. mirek z., pięko�-mirkowa h., zając a., zając m. 2002. flowering plants and pteridophytes of poland. a checklist. w. szafer institute of botany, polish academy of sciences, kraków. redhead s. a. 1975. the genus cristulariella. can. j. bot. 53: 700–707. cristulariella depraedans 11 redhead s. a. 1979. mycological observations: 1, on cristulariella; 2, on valdensenia; 3, on neolecta. mycologia 71: 1248-1253. saccardo p. a. 1908. notae mycologicae. series x. ann. mycol. 6: 553–569. saramak a. 2005. opady atmosferyczne w gaiku-brzezowej w latach 1971-2000. (in:) e. bogdanowicz, u. kossowska-cezak, j. szkutnicki (eds). ekstremalne zjawiska hydrologiczne i meteorologiczne. polskie towarzystwo geofizyczne imgw, warszawa: 184–197. skrzypczyńska m. 2004. studies on the population frequency of insects and mites causing galls on the leaves of the sycamore maple acer pseudoplatanus l. in southern poland. j. pest. sci. 77: 49–51. sydow p. 1912. mycotheca germanica. fasc. xxii–xiii. ann. mycol. 10: 445–451. trolinger j. c., elliott e. s., young r. j. 1978. host range of cristulariella pyramidalis. plant dis. reporter 62: 710–714. willetts h. j. 1997. morphology, development and evolution of stromata/sclerotia and macroconidia of the sclerotiniaceae. mycol res. 101 (8): 939–952. wulf a. 1990. über die bedeutung von diplodina acerina (pass.) sutton und anderen blattpilzen als antagonisten der fenstergallmücke dasineura vitrina kffr. an bergahorn (acer pseudoplatanus l.). nachrichtenbl. pflanzenschutzd. 42: 97–102. wulf a. 1994. pilzbedingte blattkrankheiten an ahorn unter besonderer berücksichtigung des bergahorns (acer pseudoplatanus l.). schriften aus der forstlichen fakultät der universität göttingen und der niedersächsischen forstlichen versuchsanstalt. 116: 46–58. cristulariella depreadens jako sprawca plamisto�ci li�ci klonów oraz innych drzew i krzewów s t r e s z c z e n i e badania prowadzono w okresie 1996-2006 na terenie ojcowskiego pn, tatrzańskiego pn, nadl. świerklaniec i w lesie wolskim koło krakowa (tab. 1). w okresie od lipca do października, z 30 powstałych zwykle w wyniku samosiewu młodych drzew oraz dolnych partii koron drzew starszych acer pseudoplatanus i a. platanoides pobierano 1 – 3-krotnie 30 do 60 li�ci wykazujących różne objawy chorobowe oraz po 3 do 6 li�ci z objawami chorobowymi dwudziestu innych gatunków drzew i krzewów rosnących w sąsiedztwie badanych klonów (tab. 2). w laboratorium dokonywano analizy nekroz i oznak etiologicznych c. depraedans. c. depraedans stwierdzony został we wszystkich czterech rejonach, gdzie pobierano materiał do badań, przy czym na terenie nadle�nictwa świerklaniec c. depraedans występował tylko sporadycznie. na terenie tatrzańskiego pn, nadl. świerklaniec i w lesie wolskim c. depraedans stwierdzono tylko na li�ciach acer pseudoplatanus, natomiast w ojcowskim pn na obu badanych gatunkach klonów oraz na 8 innych gatunkach drzew i krzewów: carpinus betulus, cornus sanguinea, corylus avellana, fagus sylvatica, lonicera xylosteum, padus avium, quercus robur i tilia cordata. na 12 innych gatunkach drzew i krzewów rosnących w sąsiedztwie porażonych klonów nie stwierdzono objawów porażenia przez c. depraedans (tab. 2). występowały pewne różnice w objawach porażenia zarówno pomiędzy a. pseudoplatanus i a. platanoides, jak i pomiędzy pozostałymi gatunkami ro�lin. na jednym li�ciu a. pseudoplatanus występowało od 9 do 438 nekroz. osiągały one do 22 mm �rednicy (tab. 3). pojedyncze nekrotyczne plamy były prawie okrągłe, w czę�ci �rodkowej szarobiałe, w czę�ci przyobwodowej ciemnoszare (tab. 2; figs 1, 2). na jednym li�ciu a. platanoides występowało do 338 lokalnych nekroz (tab. 3). oprócz nekroz małych 1 do 5 mm �rednicy, obecne były nekrozy do 33 mm �rednicy (tab. 3; fig. 3). nekrotyczne plamy miały barwę jasnoszarą do jasnobrunatnej w czę�ci centralnej, oraz ciemnoszarą lub brunatnoszarą w strefie graniczącej z tkankami żywymi. li�cie pozostałych gatunków ro�lin porażone były przez c. depraedans w różnym stopniu. najwięcej nekrotycznych plam stwierdzono na li�ciach cornus sanguinea i quercus robur, odpowiednio do 582 i do 286 plam (figs 7, 9). średnica nekrotycznych plam u ro�lin spoza rodzaju acer wahała się w granicach od 1 do 6 mm (tab. 3). u wszystkich gatunków ro�lin 12 t. kowalski and cz. bartnik nekrotyczne tkanki wykazywały mniej lub bardziej wyraźne różnice w zabarwieniu pomiędzy czę�cią �rodkową i przyobwodową (tab. 3). propagule miały postać kulistej lub lekko spłaszczonej względnie nieco wydłużonej, wielokomórkowej główki o �rednicy 60 do 140 µm osadzonej na septowanym, hyalinowym trzonku o długo�ci 120–180 µm i grubo�ci 10–15 µm. figs 1-4. 1. a young sycamore (a. pseudoplatanus) with symptoms of infection by c. depraedans on all leaves in the crown; 2. a leaf of a. pseudoplatanus with numerous necrotic spots caused by c. depraedans; 3. a leaf of a. platanoides with numerous necrotic spots caused by c. depraedans; 4. a leaf of a. platanoides with the local lesions of different size resulting from different infection times by c. depraedans. 1 3 2 4 figs 5-9. 5. propagules of c. depraedans on the underside of a leaf lesion on a. platanoides; 6. propagulas of c. depraedans (magnified); 7. leaves of quercus robur with symptoms of infection by c. depraedans; 8. leaf developed on sprout of carpinus betulus with symptoms of infection by c. depraedans; 9. leaf of cornus sanguinea with symptoms of infection by c. depraedans. 5 7 8 9 6 2014-01-01t11:46:53+0100 polish botanical society remarks to the ecology of the boreo-montane polypore amylocystis lapponica based on data from the czech republic and poland jan holec1 and tomáš kučera2 1national museum, mycological department, václavské nám. 68, cz-115 79 praha 1, jan_holec@nm.cz 2institute of systems biology and ecology, academy of sciences of the czech republic na sádkách 7, cz-370 05 české budějovice, kucera@usbe.cas.cz h o l e c j., k u č e r a t.: remarks to the ecology of the boreo-montane polypore amylocystis lapponica based on data from the czech republic and poland. acta mycol. 42 (2):161-168, 2007. in the czech republic, the rare polypore amylocystis lapponica (romell) singer continuously occurs in the boubínský prales virgin forest (southern bohemia: šumava mts) where it has been documented for more than 60 years. similarly, in poland it has been known only from the puszcza białowieska virgin forest (northeastern poland) for more than 50 years. generally, it is considered a species of boreal coniferous forests of northern europe (taiga) and montane coniferous forests in central and southern europe. however, the data from the czech republic, slovakia and western ukraine show that it also grows in mixed montane forests composed mainly of fagus sylvatica, picea abies and abies alba. in poland, the locality is situated in a lowland forest consisting mostly of carpinus betulus, quercus robur and tilia cordata. in central europe, a. lapponica occurs only in virgin forest refuges with the following habitat conditions: vegetation continuity (never cut), natural tree species composition, multiaged structure, rich presence of dead wood in various stages of decay, relatively large area of the virgin forest surrounded by near-natural forests, stable, cold and humid mesoand microclimate. consequently, a. lapponica may be considered an indicator of long-term vegetation continuity and stable habitat conditions. key words: fungi, central europe, habitats, virgin forests, vegetation continuity introduction amylocystis lapponica (romell) singer (basidiomycetes, polyporales, fomitopsidaceae) is a polypore typical of northern boreal forests (taiga) (r y v a r d e n , g i l b e r t s o n 1993; d a h l b e r g , c r o n e b o r g 2003). in europe, the species has a boreo-montane distribution and is extremely rare outside boreal forests. this also relates to its occurrence in the czech republic (cr) and poland (pl). in each of these countries, a. lapponica occurs only at one large locality, representing the bestpreserved virgin (primeval) forest of the country: boubínský prales (national nature reserve) and its vicinity in the mt. boubín complex (cr) and puszcza białowieska acta mycologica vol. 42 (2): 161-168 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 162 j. holec and t. kučera virgin forest (białowieża national park, pl). the occurrence in the czech republic was summarised by k o t l a b a (1984) and k o t l a b a et al. (1995), who used data from older publications by k o t l a b a and p o u z a r (1963) and p i l á t (1965), labels of herbarium collections and field notes. more recent collections from the boubín area were published on-line by v l a s á k (2005). in poland, the occurrence of a. lapponica was recently evaluated by p i ą t e k (2005). from the conservational viewpoint, amylocystis lapponica is considered a critically endangered species, included both in the red book (k o t l a b a et al. 1995) and red list of fungi of the czech republic (h o l e c , b e r a n 2006). consequently, it is protected by law in the country (a n t o n í n , b i e b e r o v á 1995). in poland, the species is also included in the national red list as a critically endangered species (wo j e w o d a , ł a w r y n o w i c z 2006) and also protected by law (ł a w r y n o w i c z 2004; also see k a r a s i ń s k i 2007: http://grzyby.strefa.pl/chronione.html). recently, d a h l b e r g and c r o n e b o r g (2003) summarised data on habitats of amylocystis lapponica in europe. unfortunately, the habitats present in the boubínský prales and puszcza białowieska virgin forests are not mentioned. the aims of this paper are (1) to summarise older data and publish new data on the occurrence of amylocystis lapponica in the boubínský prales virgin forest, (2) to compare the habitats of a. lapponica in the czech republic and poland, and (3) to add data on habitat preferences of a. lapponica in these countries to the incomplete habitat characteristics published by d a h l b e r g and c r o n e b o r g (2003). material and methods the search for amylocystis lapponica in the boubínský prales virgin forest by j. holec was not intensive and not focused only on this species. it was carried out as part of a study of mycobiota in the šumava mts in the years 1996-2006 (see e.g. h o l e c 2000; h o l e c , b e r a n 2007). consequently, his find of a. lapponica is rather accidental. however, when combined with published and herbarium data and provided with a phytosociological analysis, it can be used for an ecological synthesis. this paper is a reply to a request from the agency for nature conservation and landscape protection of the czech republic for monitoring protected species of fungi. the habitat of the recent stand of amylocystis lapponica is characterised by a phytosociological relevé. the relevé was made according to the braun-blanquet phytosociological approach with the cover being estimated by using the extended braun-blanquet scale: r (rare), + (less then 1 % cover), 1 (2-4 %), 2m (5 %), 2a (6-15 %), 2b (16-25 %), 3 (26-50 %), 4 (51-75 %), 5 (76-100 %). nomenclature of vascular plants follows k u b á t et al. (2002). voucher specimens are deposited in prm (national museum, prague, czech republic) and brnu (masaryk university, brno, czech republic). abbreviations: bpvf: boubínský prales virgin forest, cr: czech republic, pl: poland. remarks to the ecology of amylocystis lapponica 163 results amylocystis lapponica, occurrence and ecology in the czech republic geographic position of the locality. czech republic, southern bohemia, former prachatice district, šumava mts (=bohemian forest, böhmerwald), šumava protected landscape area, 9 km nw of the town of volary, mt. boubín complex: boubínský prales virgin forest (bpvf) and adjacent slopes of basumský hřbet ridge and mt. červený vrch. habitat of boubínský prales. mixed montane forest composed of fagus sylvatica, picea abies and abies alba, with admixed acer pseudoplatanus and ulmus glabra, phytosociologically a mosaic of herb-rich beech forests, montane sycamore beech forests, acidophilous beech forests and montane spruce forests with calamagrostis villosa (a l b r e c h t et al. 2003), with some trees aged 400-500 years. the core area (46.67 ha) has never been cut nor managed by foresters. it represents a remnant of original natural vegetation, with a high amount of dead and fallen decaying trunks. it has been protected as a nature reserve since 1858. forest surrounding the core area is also natural. the total area of the reserve is 677 ha, its altitude ranges from 874 to 1362 m, the core area (enclosed by a palisade) from 930 to 1100 m. coordinates of the centre of the core area are 48° 58´ 39´́ n, 13° 48´ 41´́ e. mycological characteristics of boubínský prales. see k u b i č k a (1960, 1973), partial data are present in dozens of publications on taxonomy, biodiversity and ecology of fungi both in czech and foreign literature. records of amylocystis lapponica in the boubínský prales. for details see table 1. on mt. boubín, the species is known from several microlocalities. most of them are situated in the core area (enclosed by a palisade, see below) of the boubínský prales national nature reserve. the exact geographic position of older finds by j. herink and j. kubička is not known, but it is highly probable that they originate from the core area of bpvf which was traditionally visited by mycologists at that time. however, finds of a. lapponica are known from more microlocalities in the mt. boubín complex. the exact location of finds by v l a s á k (2005) is not clear from data published by him but it is probable that those indicated by a question mark in table 1 are from bpvf (core area enclosed by a palisade plus surrounding parts of the national nature reserve). however, vlasák mentions also two finds from mt. červený vrch which is situated in the mt. boubín complex but approximately 3.5 km s of the summit of mt. boubín and outside of the boubínský prales national nature reserve. there are also nearnatural forests on slopes of mt. červený vrch which resemble those in the bpvf. the well-documented find by holec published here (see below) is situated in the national nature reserve but outside the core area. good colour photographs of a. lapponica from boubínský prales were published by p a p o u š e k (2004: no. 237). habitat conditions of the recent find by j. holec. boubínský prales national nature reserve, approximately 100 m wsw of the boubínské (kaplické) jezírko water reservoir, 10 m s of the palisade enclosing the core area close to the forest path (nature trail) around the core area, ene slope (for details on location and substrate, see table 1). 164 j. holec and t. kučera t a b le 1 su m m ar y of r ec or ds o f a m yl oc ys tis la pp on ic a in m t. b ou bí n co m pl ex ( šu m av a m ts , c ze ch r ep ub lic ) vo uc he r sp ec im en lo ca lit y m ic ro lo ca lit y su bs tr at e pa rt al ti tu de da y m on th ye ar le g. p r m 8 07 43 0, p r m 9 30 47 0 b p v f ? ? ? ? 12 9 19 46 j. h er in k p r m 6 18 50 4 m t. b ou bí n ? p ic ea ? a bi es ? de ca ye d tr un k ? 4 7 19 54 j. k ub ič ka p r m 8 70 78 3 b p v f co re a re a (e nc lo se d by pa lis ad e) p ic ea a bi es fa lle n tr un k ? 28 9 19 67 z . p ou za r p r m 6 84 42 0 b p v f co re a re a (e nc lo se d by pa lis ad e) p ic ea a bi es fa lle n tr un k ? 28 9 19 67 z . p ou za r ? b p v f ? p ic ea a bi es ? ? 19 9 19 81 j. v la sá k he rb . v la sá k m t. b ou bí n m t. č er ve ný v rc h in m t. b ou bí n co m pl ex p ic ea a bi es ? 9 19 81 j. v la sá k he rb . v la sá k b p v f ? p ic ea a bi es ? 9 19 82 j. v la sá k he rb . v la sá k b p v f ? p ic ea a bi es ? 9 19 83 j. v la sá k ? (p ho to gr ap h) ? p ic ea a bi es lo g of a fa lle n de ca yi ng tr un k 10 50 9 10 19 88 j. v la sá k ab se nt b p v f co re a re a (e nc lo se d by pa lis ad e) ? ? ? 25 10 19 90 fo un d by f . k ot la ba , s ee n in th e fie ld b y j. h ol ec he rb . v la sá k m t. b ou bí n m t. č er ve ný v rc h in m t. b ou bí n co m pl ex co ni fe r 10 19 94 j. v la sá k he rb . v la sá k b p v f ? p ic ea a bi es 9 20 01 j. v la sá k b r n u : d vo řá k 17 0/ 03 b p v f co re a re a (e nc lo se d by p al is ad e) ; n 4 8° 58 ´2 8. 2´́ e 1 3° 49 ´0 0. 6´́ p ic ea a bi es fa lle n tr un k, w it ho ut b ar k bu t w it h ra th er h ar d w oo d ? 27 9 20 03 d . d vo řá k p r m 8 98 86 7 (j h 2 35 /2 00 4) b p v f s of c or e ar ea , cl os e to p al is ad e; n 4 8° 58 ´2 2. 44 ´́ e 1 3° 49 ´0 3. 44 ´́ p ic ea a bi es fa lle n de ca yi ng tr un k (d ia m et er 4 0 cm ), fe lle d at ba se , w it ho ut ba rk , w it h so ft w oo d 95 0 9 10 20 04 j. h ol ec remarks to the ecology of amylocystis lapponica 165 phytosociological relevé (made by t. kučera 13 june 2007): total area 400 m2 (20 x 20 m), alt. 950 m, ene slope, inclination 15 °. tree layer (e3) – total cover 50 %: picea abies 3+. shrub layer (e2) – total cover 10 %: fagus sylvatica 1+, sorbus aucuparia +, picea abies 1. herb layer (e1) – total cover 15 %: vaccinium myrtillus 1+, prenanthes purpurea 1, dryopteris dilatata 1, oxalis acetosella +, luzula luzuloides +, carex pilulifera +, polygonatum verticillatum +, athyrium filix-femina +, rubus idaeus +, senecio ovatus +, lycopodium annotinum +, gymnocarpium dryopteris r, abies alba juv. r. moss layer (e0) – total cover 5 %: polytrichum commune 1, dicranum scoparium +, d. undulatum +. species outside the relevé: actaea spicata, cardamine impatiens, carex sylvatica, chaerophyllum hirsutum, circaea alpina, dentaria enneaphyllos, galeobdolon montanum, galium odoratum, impatiens noli-tangere, myosotis nemorosa, petasites albus, stellaria nemorosa. the plant community belongs to the association dentario enneaphylli-fagetum oberdorfer ex w. et a. matuszkiewicz 1960, the neighbouring stands to the subassociation impatientetosum (h a r t m a n n , j a h n 1967) m o r a v e c 1974. it falls under n a t u r a 2000 habitat 9130, asperulo-fagetum beech forests (c h y t r ý et al. 2001). this is a potential natural vegetation type, which would be present if the site were free of human influence. in fact, the site is a high-grown (approximately 100 years old) man-influenced forest with dominance of picea abies and admixed fagus sylvatica. it is not a virgin forest (trees are of the same age, dominance of picea instead of fagus, low amount of dead wood). however, natural forest almost untouched by man (virgin forest) is situated at 10 m distance (behind the palisade enclosing the core area of the nature reserve). discussion occurrence in the czech republic. to the present date, boubínský prales virgin forest is the only locality of amylocystis lapponica in the czech republic. the species has not been found in other virgin forests of the country (e.g. žofínský prales in south-eastern bohemia, mionší, salajka and razula in north-eastern moravia) in spite of a long and intensive study of polypores in the czech republic (e.g., p i l á t 1936-1942; k o t l a b a 1984; k o t l a b a , p o u z a r , va m p o l a in h o l e c , b e r a n 2006). the data document continuous occurrence of amylocystis lapponica on mt. boubín for more than 60 years. the vitality of the local population is illustrated by the fact that the species is known from several microlocalities. the following habitat conditions (which are unique in the central european landscape) seem to be decisive: vegetation continuity (never cut), natural tree species composition, multi-aged structure, rich presence of dead wood in various stages of decay, relatively large area of the virgin forest surrounded by near-natural forest (the importance of larger reserves for maintaining sensitive species was stressed by s i i t o n e n et al. 2005), and a stable, cold and humid mesoand microclimate. as it was illustrated in another paper (h o l e c , b e r a n 2007), these ”virgin forest conditions” of bpvf enable the occurrence of other fungi preferring natural forests (e.g., phellinus pouzarii kotlaba, phellinus ferrugineofuscus (p. karst.) bourdot et galzin, pseudoplectania melaena (fr.) sacc., baeospora myriadophylla (peck) singer, rigidoporus crocatus (pat.) ry166 j. holec and t. kučera varden, multiclavula mucida (pers.) r.h. petersen; see h o l e c 2003, h o l e c , b e r a n 2006). some rare species occurring in bpvf have a boreo-montane distribution (e.g., laurilia sulcata (burt) pouzar, phellinus ferrugineofuscus (p. karst.) bourdot et galzin, perenniporia subacida (peck) donk). all these data show the uniqueness of the habitat conditions of bpvf, which is the best-preserved montane virgin forest of the czech republic. occurrence in poland. in poland, amylocystis lapponica is known only from the puszcza białowieska virgin forest in north-eastern poland (p i ą t e k 2005) where it is rather common. although p i ą t e k (2005) wrote that the last find was made in 1973, photographs of a. lapponica were taken in 2005 and 2006 (k a r a s i ń s k i 2007, s n o w a r s k i 2007), which document its continuous occurrence since its first find in 1956 (d o m a ń s k i 1959). similarly like in boubínský prales virgin forest in the czech republic, puszcza białowieska in poland is an important remnant of natural vegetation which is unique not only from the polish but also the european point of view. comparison of czech and polish habitats. the following habitats of amylocystis lapponica in the puszcza białowieska virgin forest are given by p i ą t e k (2005): vaccinio uliginosi-pinetum kleist 1929, tilio cordatae-carpinetum betuli tracz. 1962, and the grouping association querco-carpinetum medioeuropaeum tüxen 1936. in all cases, the species was found on fallen trunks or branches of picea abies. puszcza białowieska virgin forest is a lowland forest (mean altitude 170 m), where spruce grows in wet depressions, the microclimatological conditions are colder than in the surrounding. in the czech republic, the locality is surrounded by the natural montane sprucefir beech forests of the very large coniferous complex of the šumava mts (montane spruce forests in upper, and spruce cultural forests in lower position, dozens of square kilometres). in contrast, the polish locality is situated in a lowland deciduous forest complex with a higher proportion of pine and spruce forests in wet depressions near the bog. therefore both localities are especially similar in their historical management (minimal influence by man) rather than habitat. remarks to habitats and occurrence in europe. d a h l b e r g and c r o n e b o r g (2003) compiled the following data on habitats of amylocystis lapponica in europe: western taiga (natura 2000 priority habitat 9010) and fennoscandian herb-rich forests with picea abies (9050) in nordic countries and estonia, acidophilous picea forest of the montane to the alpine levels (9410) in slovenia and alpine larix decidua and/or pinus cembra forests (9420) in italy. it is clear that habitats known from the czech republic and poland are not included. these differ e.g. by a high percentage of broad-leaved trees (esp. fagus sylvatica, carpinus betulus, quercus robur, and tilia cordata) forming the habitats asperulo-fagetum beech forests (9130, boubínský prales) and galio-carpinetum oak hornbeam forests (9170, puszcza białowieska), respectively. the number of localities in various european countries is very different. while hundreds of localities in sweden and finland and dozens in norway (d a h l b e r g , c r o n e b o r g 2003) are known, in countries of central and southern europe the species is extremely rare and mostly occupies only one locality per country. it clearly shows that localities in central and southern europe represent refuges of a. lap remarks to the ecology of amylocystis lapponica 167 ponica outside the zone of boreal forests of northern europe. these refuges are formed by azonal soil or topoclimatic conditions in the temperate forest zone. in northern europe, amylocystis lapponica is considered as a good indicator of old-growth and species-rich spruce forests which are in special need of conservation (norway: b r e d e s e n et al. 1997; r ø s o k 1998; finland: s i i t o n e n et al. 2005). however, j o n s s o n and j o n s e l l (1999) showed in swedish spruce forests that if indicator species are to be used for evaluation of their biodiversity, these should be chosen from several groups of organisms (e.g. vascular plants, bryophytes, epiphytic lichens and wood-inhabiting fungi). this is certainly true but at least in central europe, a. lapponica indicates high continuity of forest vegetation which has never been cut. supporting data from the czech republic and poland are summarised here. data from slovakia (k o t l a b a , p o u z a r 1963; p i l á t 1965; k o t l a b a 1984; va r j ú 1994; š k u b l a 2003) and eastern carpathians in western ukraine (p i l á t 1936-1942, 1940; for current names of his localities, see h o l e c 2002) are almost identical. amylocystis lapponica occurs there exclusively in virgin forests (central slovakia: dobročský prales, voucher specimens in prm and herb. vlasák; eastern carpathians: altogether 19 collections from several localities, voucher specimens in prm). these forests are composed mainly of fagus sylvatica, abies alba and picea abies and have the same „virgin forest character“ as was specified in the first chapter of the discussion. acknowledgements. we thank daniel dvořák (masaryk university, brno, czech republic) for providing data on his collection of a. lapponica. jan holec was financially supported by the ministry of culture of the czech republic (mk00002327201), tomáš kučera by institutional research plan av0z60870520. references a n t o n í n v., b i e b e r o v á z. 1995. chráněné houby čr (fungi protected by law in the czech republic). ministerstvo životního prostředí čr, praha. b r e d e s e n b., h a u g a n r., a a n d e r a a r., l i n d b l a d i., ø k l a n d b., r ø s o k ø. 1997. woodinhabiting fungi as indicators on ecological continuity within spruce forests of southeastern norway. blyttia 54: 131–140. da h l b e r g a., c r o n e b o r g h. 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(in:) k. k a v i n a , a. p i l á t (eds). atlas des champignons de l’europe. 3. praha. p i l á t a. 1940. hymenomycetes carpatorum orientalium. sborn. nár. mus. v praze, řada b, přír. vědy. acta musei nationalis pragae, ser. b. 2 (3): 37–80. p i l á t a. 1965. modralka laponská – amylocystis lapponica (romell) bond. et sing. čes. mykol. 19: 9-10, tab. 56. r ø s o k ø. 1998. amylocystis lapponica in norway, an indicator species for species-rich continuity forests. blyttia 56: 154-165. r y v a r d e n l., g i l b e r t s o n r. l. 1993. european polypores. part 1. fungiflora, oslo. s i i t o n e n p., l e h t i n e n a., s i i t o n e n m. 2005. effects of forest edges on the distribution, abun-effects of forest edges on the distribution, abundance, and regional persistence of wood-rotting fungi. conserv. biol. 19 (1): 250–260. s n o w a r s k i m. 2007. atlas grzybów polski. fungi of poland. wersja 2007.05.27w. http://www.grzyby.pl/ (accessed 18 june 2007). š k u b l a p. 2003. mycoflora slovaca. mycelium, šaľa. va r j ú l. 1994. zoznam trúdnikotvarých húb nájdených v dobročskom pralese 9.-10. októbra 1993. spravodajca slovenských mykológov 2 (3): 25. v l a s á k j. 2005. polypores. collection of dr. josef vlasák, hluboká nad vltavou, czech republic. edition xi.05. http://botanika.bf.jcu.cz/mykologie/polypores/ (accessed 15 june 2007). w o j e w o d a w., ł a w r y n o w i c z m. 2006. red list of the macrofungi in poland. (in:) z. m i r e k , k. z a r z y c k i , w. w o j e w o d a , z. s z e l ą g (eds). red list of plants and fungi in poland. w. szafer institute of botany, polish academy of sciences, kraków: 53–70. 2014-01-01t11:45:56+0100 polish botanical society 2014-08-20t22:54:02+0200 polish botanical society 2014-08-20t22:46:21+0200 polish botanical society 2014-08-20t21:45:20+0200 polish botanical society 2014-08-25t21:39:29+0200 polish botanical society europe, a continent with high potential for the cultivation of the burgundy truffle (tuber aestivum/uncinatum) gérard chevalier résidence cristelle, 33 rue des vergers, f-63800 cournon d’auvergne, charlemagne33@orange.fr chevalier g.: europe, a continent with high potential for the cultivation of the burgundy truffle (tuber aestivum/uncinatum). acta mycol. 47 (2): 127–132, 2012. the burgundy truffle (tuber aestivum/uncinatum) grows, in europe, in the most varied soils from a physical and chemical viewpoint. the only common point is the presence of a minimum level of exchangeable calcium in the soil. the truffle soils in europe can be classified in two categories: the soils coming directly from the parent rock, and those formed from deposits covering the parent rock. the first group corresponds to most traditional truffle areas , france, italy and spain. it is also true for some soils from ireland to eastern europe (romania, bulgaria), from southern europe (greece, former yugoslavia) to northern europe (sweden, baltic countries). the sedimentary layers that cover most areas are from the secondary and tertiary era. the primary parent rocks are less frequent. the second group means quaternary or recent alluviums covering the parent rock on huge surfaces and at great depth sometimes. they characterize mainly countries eastern and northern europe. by bringing within reason limestone, it is possible to cultivate t. aestivum/uncinatum on sedimentary soils non-calcareous or decalcified or even on soils issued from magmatic rocks (granite) or metamorphic (schists). the possibilities of truffle cultivation in europe are therefore enormous, the limiting factors not being the soil but the climate. key words: truffles, soil, calcium, liming, truffle cultivation introduction hundreds of soil analyses carried out from the 1970’s show that the burgundy truffle (tuber aestivum/uncinatum) grows, in europe, in the most varied soils from a physical and chemical viewpoint, far more than the soils for t. melanosporum and even more for t. magnatum. the only common point is the presence of a minimum level of exchangeable calcium in the soil (chevalier 2011). the nature of the parent rock conditions the soil physico-chemical characteristics. the soils derived from calcareous parent rock, named calcimagnesic soils acta mycologica vol. 47 (2): 127–132 2012 128 g. chevalier or calcimorphic soils are “intrazonal” soils, that are linked more to the nature of the parent rock they were derived from, than the climatic zone where they reside (mathieu 2009). under humid temperate climate, the evolution of the calcareous parent rock is linked to the decarbonatation of this rock. truffle soils the truffle soils in europe can be classified in two vast categories: the soils coming directly from the parent rock, and those formed from deposits covering the parent rock (fig.1). the first group corresponds to the most traditional truffle areas, france (chevalier 2008, 2010a; chevalier, frochot 1997; chevalier, palenzona 2011; chevalier et al. 1979) (fig.2), italy (bencivenga 2008; granetti et al. 2005; gregori 1991, 2010; raglione et al. 2011; zambonelli, di munno 1992) and spain (de miguel et al. 2008; honrubia et al. 2011). it is also true for some soils in the united kingdom (french 2008) , ireland, germany (southern half) (fig.3), switzerland (groux, keller 2008), austria (urban, pla 2008), czech republic, slovakia (gazo, miko 2008, 2010), southern poland (hilszczańska, sierota 2010; lawrynowicz 2011) (fig.4), hungary (bakony mountains) (bratek 2008; bratek et al. 2010) , romania (carpathian mountains), bulgaria, greece, former yugoslavia (marjanovic 2008), sweden (isles of gotland and öland) (wedén, danell 2010; wedén et al. 2004), baltic republics. the sedimentary layers that cover most areas are from the secondary and tertiary era. the primary parent rocks are less frequent: ireland, northern united kingdom, the ardennes and eifel hills, rhenish schist massif, the harz, southwest of saxony and southeast of thuringia, bohemia, southern poland (silesia, katowice region, lysa gora), swedish isles, baltic republics. the second group means quaternary or recent alluviums covering the parent rock on huge surfaces and at great depth sometimes. they characterize the po valley, southern germany (glacial till), the austrian, slovak, hungarian and romanian plains. large areas also are found above the glacial line showing the maximal advance of the quaternary glaciers. this is valid for the northern half of the netherlands, denmark, northern germany (fig. 3), poland (fig. 4), belarus and ukraine. in this second group the alluviums which cover the parent rock are calcareous or calcic (loess) and the truffle can grow; the alluvium are free of limestone or calcium and the truffle fails to develop, even though the parent rock may be calcareous but deep. by bringing within reason limestone, it is possible to cultivate t. aestivum/uncinatum on sedimentary soils non-calcareous or decalcified or even on soils issued from magmatic tocks (granite) or metamorphic (schists) (such a cultivation in france from the 2000’s) (chevalier 2003, 2010b). the sedimentary soils of this type cover large surfaces in northern europe, north of the glacier most advance demarcation. europe ... for the cultivation of truffle 129 fig.1.geologic map of europe. the northern part is characterized by soils originated from deposits covering the parent rock (in beige and grey). the soils of the southern part are more various. southern limit of the quaternary glaciation (black line in dash). fig. 2. truffle soils of france. the truffle soils are very various. many of them are suitable for the growth of t. uncinatum. only some soils of the west (bretagne), central massif, east (vosges) and south (pyrénées and corse) are unsuitable without liming (in red, violet and purple). 130 g. chevalier fig. 3. truffle soils of germany. germany presents the two types of soils: deposits covering the parent rock in the north (in beige and grey); soils originated from breaking up of the parent rock in the south. the soils of the southern part are more suitable for truffles. fig. 4. truffle soils of poland. the great poland plain is covered by quaternary deposits from glacier or wind origin. the soils of the southern poland are quite different and closer to those of southern europe but their surface is more limited. they are originated from the breaking up of the parent rock that can be jurassic (north-west of krakow, in blue), cretaceous (beskides, south and south-east of lublin, in green) or tertiary (polish carpathians). the areas of the southern poland are much more favourable to the truffles than those of the great plain. europe ... for the cultivation of truffle 131 conclusion the possibilities of cultivation in europe are therefore enormous, the limiting factor not being the soil but the climate (considerable heat in summer, summer drought, and severe cold in winter). preempting the cold, the heat, drought is possible by adaptive cultivation methods (in depth work of the soil) and the temporary cover of the soil (mulching). the damaged caused by the drought can also be mitigated by a controlled irrigation. references bencivenga m. 2008. la coltivazione dei tartufi in italia. (in:) le causse corrézien (eds). la culture de la truffe dans le monde: 125–134. brive. bratek z. 2008. truffe et plantations à vocation truffière en hongrie. (in:) le causse corrézien (eds.). la culture de la truffe dans le monde, brive: 95–109. bratek z., merenyi z., illies z., laslo p., anton a., papp l., merkl o., garay j., vikor j., brandt s. 2010. studies on the ecophysiology of tuber aestivum populations in the carpatho-pannonian region. österr. z. pilz. 19: 221–226. chevalier g. 2003. aquitaine: des truffes dans le limousin! le trufficulteur français 43: 9. chevalier g. 2008. la trufficulture en france. (in:) le causse corrézien (eds). la culture de la truffe dans le monde, brive: 59–77. chevalier g., 2010a. la truffe d’ europe (tuber aestivum): limites géographiques, écologie et culture. österr. z. pilzk. 19: 249–259. chevalier g. 2010b. la m.r.t. (méthode raisonnée de trufficulture). (in:) office de tourisme de sarlat (eds). les nouvelles techniques de culture de la truffe: 55–67. sarlat. chevalier g. 2011. le calcium, clé de la production de tuber aestivum/uncinatum. (in:) truffe 54 lorraine (eds). proc. 3d congress of the european scientific group tuber aestivum/t. uncinatum (t.a.u.e.s.g.), nancy, nov. 7th 2011 (in press). chevalier g., frochot h. 1997. la truffe de bourgogne (tuber uncinatum chat.): histoire, biologie, écologie, culture, récolte, gastronomie. pétrarque, levallois-perret. chevalier g., palenzona m. 2011. truffes et trufficulture en europe. (in:) comunita montana dei monti martani e del serano (eds). proc. 3d intern. congr. on truffle, spoleto: 65–72. chevalier g., desmas c., riousset l.g. 1979. l’ espèce tuber aestivum vitt. ii. ecologie. mushr. sc. 10 (1): 977–993. de miguel a., reyna s., saez t., palazon c., 2008. la trufficulture en espagne. (in:) le causse corrézien (eds). la culture de la truffe dans le monde: 45–58. brive. french m.a. 2008. le développement du marché truffier en grande-bretagne. (in:). le causse corrézien (eds) : la culture de la truffe dans le monde: 79–93. brive. gažo j., miko m. 2008. history and perspectives of economical use of the burgundy truffles (tuber aestivum vitt.) in slovak republic. (in:) le causse corrézien (eds). la culture de la truffe dans le monde: brive: 173–182. gažo j., miko m. 2010. truffle habitat in western slovakia. österr. z. pilzk. 19: 245–247. granetti b., de angelis a., materozzi g. 2005. umbria, terra di tartufi. regione umbria, terni. grebenc, t., kraigher h., martin mp., piltaver a., ratosa i. 2008. research and cultivation of truffle in slovenia – current status. (in:) le causse corrézien (eds). la culture de la truffe dans le monde, brive: 183–191. gregori g. 1991. tartufi e tartificoltura nel veneto. assessorato agricoltura e foreste, padova. gregori g. 2010. l ‘espèce tuber uncinatum en italie : écologie, commercialisation et valorisation. österr. z. pilzk. 19: 265–271. groux m., keller j., 2008. la truffe en suisse. (in:) le causse corrézien (eds). la culture de la truffe dans le monde, brive: 209–217. 132 g. chevalier hilszczańska d., sierota z. 2010. first attempts towards cultivation of tuber aestivum in poland. österr. z. pilzk. 19: 209–212. honrubia m., alfaro r., fernandez a., suz l.m., morte a. 2011. location and ecological characterization of natural truffle grounds in the region of murcia (south east of spain). (in:) comunita montana dei monti martani e del serano (eds). proc. 3d intern. congr. on truffle, spoleto: 189. lawrynowicz m. 2011. hypogeous fungi in the anthropogenic sites in poland. (in:) comunita montana dei monti martani e del serano (eds). proc. 3d intern. congr. on truffle, spoleto: 166–172. marjanovic z. 2008. truffles and possibilities for their cultivation in serbiacurrent situation. (in:) le causse corrézien (eds). la culture de la truffe dans le monde, brive: 163–172. mathieu c. 2009. les principaux sols du monde . voyage à travers l’ épiderme vivant de la planète terre. tec & doc, lavoisier, paris. raglione m., lorenzini p., malgorzata o., bonifazi a. 2011. a contribution to the characterization of the soils suitable for tuber melanosporum and t. aestivum growth through mineralogical analysis of clay fraction. (in:) comunita montanadei monti martani e del serano (eds). proc. 3d intern. congr. on truffle, spoleto: 147–151. urban a., pla t. 2008. truffles and truffle cultivation in austria. (in:) le causse corrézien (eds). la culture de la truffe dans le monde, brive: 19–34. wedén c., danell e. 2010. truffle cultivation in sweden. (in:) le causse corrézien (eds). la culture de la truffe dans le monde, brive: 193–207. wedén c., chevalier g., danell e. 2004. tuber aestivum (syn.t. uncinatum) biotopes end their history on gotland, sweden. mycol. res. 108: 304–310. zambonelli a., di munno r. 1992. indagine sulla possibilità di diffusione dei rimboschimenti con specie tartufigene: aspetti technico-colturali and economici. ecoplanning. 2014-01-02t12:09:00+0100 polish botanical society 2014-08-27t16:49:04+0200 polish botanical society 2014-08-26t20:32:28+0200 polish botanical society © the author(s) 2014 published by polish botanical society leucocoprinus lilacinogranulosus (henn.) locq. in poland błażej gierczyk1 and grzegorz dubiel2 1faculty of chemistry, adam mickiewicz university in poznań umultowska 89b, pl-61-614 poznań, hanuman@amu.edu.pl 2fałata 2d/2, pl-43-360 bystra, gdbb@interia.eu gierczyk b., dubiel g.: leucocoprinus lilacinogranulosus (henn.) locq. in poland. acta mycol. 49 (1): 59–67, 2014. the paper presents the first finding of leucocoprinus lilacinogranulosus in poland, the rare, exotic fungus, sporadically found in europe. the species was found indoors, on soil in terrarium. a description based on the specimens from poland as well as photographs and microcharacter drawings are presented. key words: basidiomycota, diversity, distribution, alien species introduction the genus leucocoprinus pat. (agaricales, basidiomycota) groups the saprotrophic and terrestrial fungi, growing on soil, woodchips, wood or compost heaps. they form more or less fragile, lepiotoid to almost coprinoid basidiomata, characterized by striate pileus (at least in marginal zone), free lamellae, annulus and pale spore print. the leucocoprinus spores are thick-walled, dextrinoid. the basidia are surrounded by pseudoparaphyses. cheilocystidia are present, while the pleurocystidia do not occur. clamps are absent in all parts of basidiomata (vellinga 2001). most of the leucocoprinus species are of the tropical origin, and have been introduced to temperate zone with tropical plants (vellinga 2001; lange 2008). depending on the systematic treatment, hitherto 19 representatives of this genus have been found in europe (canduso, lanzoni 1990; ludwig 2012b). most of them occur indoors, in buildings and greenhouses, where they grow in flower-pots. some species, e.g. l. cepistipes, acclimatize and spread, so they could be found outdoors (vellinga 2001; gierczyk et al. 2011). in poland five species of leucocoprinus have been found until now: l. badhamii, l. birnbaumii, l. caepistipes, l. straminellus (= l. denudatus) and l. cygneus (= sericeomyces cygneus) (wojewoda 2003; wojewoda, karasiński 2010; gierczyk et al. 2011; kujawa 2012). the most common is l. birn­ baumii. acta mycologica vol. 49 (1): 59–67 2014 doi: 10.5586/am.2014.005 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 60 b. gierczyk and g. dubiel © the author(s) 2014 published by polish botanical society in 2011 leucocoprinus lilacinogranulosus (henn.) locq., the species new to poland, was found indoor, in terrarium. this paper presents the description of this interesting species based on the specimens found. materials and methods the basidiomata of l. lilacinogranulosus were studied according to standard methods used in the taxonomy of fungi. the microscopic structures were examined in dried material, mounted in congo red (1% in 10% ammonia solution) using bresser bino researcher microscope. the drawings of microcharacters were made from microphotographs taken with nikon coolpix 950 digital camera. all measurements were made directly through the microscope under an oil immersion objective (×100). the spore dimensions were established from measurements of 100 randomly selected, well formed spores (the deformed or atrophied ones were excluded from analysis). microscopic measurements are presented according to the method used by breitenbach, kränzlin (1991). the 95% population limits for the mean were calculated and the lower and upper values are given. for basidia and cystidia the extreme size values were presented. for these structures dimensions were obtained after measuring of 50 elements. the material collected was deposited in the private herbarium of one of the authors (bg). the literature dimensions of the anatomical structures were given after ludwig (2012a). results and discussion iconography and description leucoagaricus lilacinogranulosus hennings (1898): 144; pl. i: fig. 17 (as lepiota li­ lacinogranulosa); bon (1981): 67; smith, weber (1982): 298-299 & 306; pl. 1: fig. 4a-c & pl. 2: fig. 4a,b; babos (1985): 212-213; reid (1989): 418-421, fig. 5a-j (var. lilacinogranulosus & var. subglobisporus); canduso, lanzoni (1990): 484-487; fig. 107; bon (1993): 112 & 153; pl. 6a (as l. ianthinus); zuccherelli, migliozzi (1998): 197-203 (var. subglobisporus); blanco-dios (2001): 13-15, fig. 1 (var. subglobispo­ rus); salom, siquier (2001): 115-116; fig. 4; luque (2009): 1-4 (as l. ianthinus); birkebak (2010): 99-100, fig. 7 (as l. ianthinus); asef, muradov (2012): 4-5, fig. 2 (as l. ianthinus); ludwig (2012a): 552-554; ludwig (2012b): 189; pl. 580: fig. 113.8. leucoagaricus ianthinus: cooke (1883): 363; cooke (1888): 101; cooke (1898): pl. 944, fig. a; reid (1989): 418-419, fig. 5k-n excluded: vellinga (2001): 80-81, fig. 53 (as l. ianthinus) – description given is the sum of the characters of l. ianthinus and l. lilacinogranulosus; roux (2006): 991 (as l. ianthinus) – presented specimens has very dark fibrils on pileus, probably other species (l. heinemanni?). leucocoprinus lilacinogranulosus (henn.) locq. in poland 61 © the author(s) 2014 published by polish botanical society description of polish specimens basidiocarp small to medium size (fig. 1). pileus 10-20 mm when expanded (lit. 1550 mm), first closed, campanulate becoming broadly campanulate to flat with age, with distinct, obtuse umbo, plicate up to 2/3 of the radius, thin-fleshed. pileus covering granulose to flocculose, with distinct purplish to lilac tinge, at centre dense and felted, darker, becoming less distinct and paler in outer half. background whitish to yellowish. stipe thin and fragile, 30-40 × 1.5-2 mm (lit. 35-150 × 1.5-3 mm), with bulbous base, hollow, above annulus white to yellow, below white, with lilac fibrils at the base. lamellae moderately crowded, free, white. annulus ascending, white with a margin of the pileus covering colour, evanescent. spores thick-walled, in sideview ellipsoid to amygdaloid, sometimes indistinctly apically papillate, in frontal view ellipsoid, 8.0-11.0 × 6.2-7.5 μm (lit. 7-11 × 5.5-7.5 μm), q = 1.35-1.72, with a germ pore covered with a hyaline, domelike cap, dextrinoid, congophilous. basidia clavate, 4-spored, 20-40 × 7-12 μm, surrounded by pseudoparaphyses. lamellar edge semifertile, with basidia intermixed with subglobose, broadly fusoid to clavate cheilocystidia, 40-60 × 10-23 μm (lit. 35-65 × 10-26 μm). pleurocystidia absent. pileus covering composed from agglutinating globose, ellipsoid and broadly cylindrical sometimes diverticulate elements, forming chains, up to 25 μm in diameter. pigment intracellular. clamp connection absent (fig. 2). specimens examined: bystra village near bielsko biała city, terrarium in private apartment; atpol: df-93; 12.01.2012; leg. & det. g. dubiel, rev. b. gierczyk; few basidiocarps on peat substratum; bgf/bf/gdu/120129/0001. taxonomic remarks there are at least two very similar fungi, recognized by some mycologists as a conspecific, while by others as separate “good” species: leucocoprinus ianthinus (cooke) p. mohr and l. lilacinogranulosus (henn.) locq. the first of them was described by cooke on the basis of the specimens collected in kew gardens (great britain) as agaricus ianthinus in 1883 (cooke 1883, 1888). the author writes: “pileus rather fleshy, umbonate (...) whitish at the even margin, disc rather violet, fibrilose, rest of pileus streaked with innate radiating, violet, hair like squamules, steam (...) whitish...”. in 1898 hennings published the diagnosis of lepiota lilacinogranulosa based of findings from botanic garden in berlin (hennings 1898). this species was described as: “pileo membranaceo, centro subcarnoso (...) umbonate violaceo-brunneo granulato, radiato-straito, squamulis liliacinis tectis; stipe (...) pallido, basi bulbilloso violaceo (...) sclerotis albis tomentosis...”. cooke did not report the spore size for his collection of l. ianthinus, while according to hennings l. lilacinogranulosus specimens produced 10-13 × 7-9 μm spores. the dimensions of the spores of type specimens of the later species, determined by mohr (1994) are slightly smaller. also wasser (1993) has reported smaller spores (see table 1), however ludwig (2012b) and babos (1985) have found the scattered spores up to 14 μm long in their collections of this species. for the first time the size of spores of l. ianthinus has been given by reid (1989): 6.5-10.0 × 5.75-6.5 μm. 62 b. gierczyk and g. dubiel © the author(s) 2014 published by polish botanical society fig. 1. specimens of leucocoprinus lilacinogranulosus from poland (photo by g. dubiel; 12.01.2012). fig. 2. microcharacters of leucocoprinus lilacinogranulosus from poland: a – spores; b – cheilocystidia; c – pileus covering; d – basidia. scale bar = 10 μm. leucocoprinus lilacinogranulosus (henn.) locq. in poland 63 © the author(s) 2014 published by polish botanical society the first arguments for and against the conspecifity of both species were given by reid (1989), but the author did not present any conclusion. bon (1993) has followed this concept and synonimised them, although he speculated that the differences in spore sizes and the presence of sclerotium (in l. lilacinogranulosus) may suggest that they are separate but close related species. also the authors of some later crucial monographs and keys, e.g. flora agaricina neerlandica (vellinga 2001) or röhlinge and blätterpilze (horak 2005) have synonymised them. ludwig (2012b) has described l. lilacinogranulosus and separated it from l. ianthinus in discussion. this conception has also been accepted by migliozzi (1996). the differences between these two taxa are collected in table 1. as the molecular studies of these species have not been hitherto performed, there are no modern arguments on the thesis of their conspecifity, therefore we agree with the opinion of the mycologists, who separate this species. there is some inconsequency in the description of l. lilacinogranulosus (and/or l. ianthinus). reid (1989), knudsen (1992) and wasser (1993) have written “cheilocystida absent” while others authors of leucocoprinus and lepiotaceae monographs as well as mycological keys (e.g. ludwig 2012a or smith, weber 1982) have reported the presence of the cheilocystidia in this species. this difference may come from the shape and character of cheilocystidia – they are well formed only in young specimens and disappear with age (göger, mohr 1992) and in some collections they are similar to basidioles or not distinct (bon 1981; canduso, laznoni 1990). our collection meets better the description of l. lilacinogranulosus (henn.) locq. the basidiomata have a sulcate pileus margin and not dark violet colours of the cap covering. the spores are over 6 μm broad. the cheilocystidia are present in the specimens studied, but difficult to find in old basidiocarps. other similar species of genus leucocoprinus, which may be confused with the fungus described here is l. cepistipes (sow.) pat. which differs macroscopically mainly in the colour of pileus, which has not lilaceous or purplish tinges. the microscopic differences are more pronounced – the later species has a distinct cheilocystidia with apical excrescences and different kinds of the elements of the pileus covering (veil is composed from branched, cylindrical hyphae and contains fusoid terminal elements; the subglobose elements are absent). somewhat similar is also l. brebissonii (godey) locq. its dark coloured, contrasting with background scales may fade with age and turn to purplish-brown. hover the young basidiomata are distinctly different in colour, moreover the spores of l. brebissonii are more elongated then that of l. lilacinogranulosus and have a distinct papilla-like apex. table 1 differences between leucocoprinus lilacinogranulosus and l. ianthinus character leucocoprinus lilacinogranulosus l. ianthinus pileus membranous, with umbo and plicate margin fleshy, with umbo and even margin pileus covering pale violet-brown to lilaceous dark violet fibrils stipe whitish with violet base white to whitish spores 10-13 × 7-9 μm (hennings 1898) (7)8-10.5(11) × (5.5)6-7.5(8) μm (ludwig 2012b) 7.5-9.75 × 5-7 μm (wasser 1993) 6.5-10 × 5.75-6.5 μm (reid 1989) 64 b. gierczyk and g. dubiel © the author(s) 2014 published by polish botanical society leucocoprinus ianthinus seems to be much rarer than l. lilacinogranulosus, since only a few reports on its finding have been published (see habitat and distribution). however, due to taxonomic subtleties and different species concept, in the case of some publication it is difficult to distinguish which of the above discussed species is mentioned, especially if the authors have not given the description and photographs of the specimens found. from great britain, italy and spain a variety forming subglobse spores – l. lil­ iacinogranulosus (henn.) locq. var. subglobisporus reid has been reported (reid 1989; zuccherelli, migliozzi 1998; blanco-dios 2001). habitat and distribution in europe both species have been hitherto found only indoors, in flowerpots in buildings and greenhouses or outdoor in warmer regions, but always with cultivated plants, in gardens. l. lilacinogranulosus is widespread but everywhere rare and sporadically collected. it has been mentioned from many countries: austria (hausknecht, pidlich-aigler 2004), belgium (walleyn, vandeven 2006), czech republic (zelený 2006), france (bon 1993), germany (hennings 1898; gminder 2003), great britain (cooke 1888; reid 1989), hungary (babos 1985), italy (canduso, lanzoni 1990), liechtenstein (prongué et al. 2004), netherlands (arnolds 1984), nordic countries (knudsen 1992; ludwig 2012a), and spain (salom, siquier 2001; luque 2009). also the extraeuropean data have mentioned it from the pots and flower-beds with ornamental, exotic plants but also from semi-natural communities. l. lilacinogranulosus has been collected in contiguous united states (smith, weber 1982; birkebak 2010), hawaii (hemmes, desjardin 2002), india (kumar, manimohan 2009; the brief description given by the authors raises doubts about the correctness of species determination) and iran (asef, muradov 2012). it is recognized as an alien species in europe with category a – “alien taxon from outside europe” (desprez-loustau 2009, as l. ian­ thinus). the origin of both varieties of this species is not known, because they have been not found hitherto in nature. nominative variety of l. lilacinogranulosus as well as var. subglobisporus has not been mentioned from poland territory until now. conclusions the intensified import of ornamental exotic plants as well as different kinds of organic substrata for plant growing have caused expansion of some fungi, therefore much attention should be given to the species forming basidiomata in flower pots, in greenhouses and on flower-beds with ornamental plants. as shown (desprez-loustau 2009; wojewoda, karasiński 2010), some of them may spread out and acclimatize, moreover they may become invasive species. although leucocoprinus lilacinogranulosus is strictly stenothermal species, and there is no information about its outdoor growing in europe, the climate warming and natural selection may cause its acclimatization and penetration of natural leucocoprinus lilacinogranulosus (henn.) locq. in poland 65 © the author(s) 2014 published by polish botanical society habitats. this species, together with the others belonging to genus leucocoprinus, are often mentioned on the internet discussion boars of plant-keepers and gardeners. there are no premises that these species may disturb the growing of plants. the finding described in this article, on soil in terrarium without any plants, shows that it is a saprobic species, not a mycorhizal or parasitic one. acknowledgements. the authors thank to dr. anna kujawa (institute for agricultural and forest environment, polish academy of sciences, field station in turew) for helpful comments and to the anonymous reviewers for useful remarks on the manuscript. references arnolds e. 1984. standaardlijst van nederlandse macrofungi. coolia 26, supplement, 1-362. kumar t.k.a., manimohan p. 2009. the genera leucoagaricus and leucocoprinus (agaricales, basidio­ mycota) in kerala state, india. mycotaxon 108: 385-428. http://dx.doi.org/10.5248%2f108.385 asef m.r., muradov p. 2012. lepiotaceous fungi (agaricaceae) in the iranian part of caucasia. turk. j. bot. 36: 1-6. babos m. 1985. studies of hungarian lepiota s.l. species. vi. glasshouse species. agarica 6 (12): 197-218. birkebak j.m. 2010. the genus leucocoprinus in western washington. mycotaxon 112: 83-102. http:// dx.doi.org/10.5248%2f112.83 blanco-dios j.b. 2001. estudios sobre o xénero leucocoprinus pat. na península ibérica. i. leucocoprinus lilacinogranulosus (henn.) locq. var. subglobisporus reid. mykes 4: 13-15. bon m. 1981. clé monographique des “lépiotes” d’europe. doc. mycol. 11 (43): 1-77. bon m. 1993. les lepiotes. lepiotaceae roze. generes: cystolepiota, melanophyllum, echinoderma, le­ piota, chamaemyces, sericeomyces, leucoagaricus, leucocoprinus, macrolepiota, chlorophyllum. flora mycologique d’europe. 3: 1-153 (doc. mycologiques memoire hors serie no. 3). l’association d’ecologie et mycologie, lille. breitenbach j., kränzlin f. 1991. fungi of switzerland. 3. boletes and agarics (1st part). verlag mykologia, luzern. canduso m., lanzoni g. 1990. lepiota s.l. fungi europaei. libreria editrice giovanna biella, saronno. cooke m.c. 1883. handbook of british fungi. london cooke m.c. 1888. new british fungi. grevillea 80, 101-102. cooke m.c. 1889-1891. ilustrations of british fungi (hymenomycetes). 8. (supplement). williams & norgate, london. desprez-loustau m.-l. 2009. alien fungi of europe. (in:) daise (ed.). handbook of alien species in europe. springer, new york. gierczyk b., kujawa a., szczepkowski a., chachuła p. 2011. rare species of lepiota and related genera. acta mycol. 46 (2): 137-178. gminder a. 2003. lepiotaceae. (in:) g.j. krieglsteiner (ed.). die groβpilze baden-württembergs. 4: 48-138. verlag eugen ulmer, stuttgart. göger f., mohr p. 1992. schlüssel für in europa beobachteten faltenschirmlinge (leucocoprinus-arten). myk. mitt. bl. 35(2): 79-82. hausknecht a., pidlich-aigener h. 2004. lepiotaceae (schirmlinge) in österreich 1. die gattungen chamaemyces, chlorophyllum, leucoagaricus, leucocoprinus, macrolepiota, melanophyllum und sericeomyces. öster. z. pilzk. 13: 1-38. hemmes d.e., desjardin d.e. 2002. mushrooms of hawaii. an identification guide. ten speed press, berkeley. hennings p. 1898. die in den gewächshäusnern des berliner botanischen gartens beobachteten pilze. verhandl. bot. ver. prov. brand. 40: 109-178. horak e. 2005. röhlinge and blätterpilze in europa, 5 ed. elsevier, heidelberg. knudsen h. 1992. leucocoprinus pat. (in:) l. hansen, h. knudsen (eds). nordic macromycetes. 2: 223224. nordsvamp, copenhagen. 66 b. gierczyk and g. dubiel © the author(s) 2014 published by polish botanical society kujawa a. 2012. grzyby makroskopijne polski w literaturze mikologicznej. (in:) m. snowarski (ed.) atlas grzybów polski (http://www.grzyby.pl/grzyby-makroskopijne-polski-w-literaturze-mikologicznej. htm). version february 2012, access date 01.08.2012. lange c. 2008. leucocoprinus pat. (in:) h. knudsen, j. vesterholt (eds). funga nordica. agaricoid, boletoid and cyphelloid genera. pp. 553-554. nordsvamp, copenhagen. luque m. 2009. leucocoprinus ianthinus (cooke) p. mohr, boletus 18 (2):48 (1994). micobotánica-jaén http://www.micobotanicajaen.com/revista/articulos/mluquev/aportaciones015/leucocoprinus%20ianthinus%20%28cooke%29%20mohr.pdf ludwig e. 2012a. pilzkompendium. 3. beschreibungen. die übrigen gattungen der agaricales mit weiβem sporenpulver. fungicon-verlag, berlin. ludwig e. 2012b. pilzkompendium. 3. abbildungen. die übrigen gattungen der agaricales mit weiβem sporenpulver. fungicon-verlag, berlin. migliozzi v. 1996. un genere alla volta. 3. introduzione allo studio del genere leucocoprinus patouillard. boll. gr. micol. bres. 39 (1): 14-16 and 73-82. mohr p. 1992. anmerkungen zu einigen faltenschirmlings-arten (leucocoprinus). boletus 18 (2): 47-51. prongué j.-p., wiederin r., wolf b. 2004. die pilze des fürstentums liechtenstein. naturkundliche forschung im fürstentum liechtenstein. band 21. amtlicher lehrmittelverlag, vaduz. reid d.a. 1989. notes on leucocoprinoid fungi from britain. mycol. res. 93: 413-423. roux p. 2006. mille et un champignons. édition roux, sainte-sigolène. salom j.c., siquier j.l. 2001. contribució al coneixement de la família lepiotaceae roze a les illes baleares. ii. rev. catalana micol. 23: 109-120. smith h.v., weber n.s. 1982. selected species of leucocoprinus from the southeastern united states. contr. univ. mich. herb. 15: 297-309. vellinga e.c. 2001. leucocoprinus pat. (in:) m.e. noordeloos, t.w. kuyper, e.c. vellinga (eds). flora agaricina neerlandica. critical monographs on families of agarics and boleti occurring in the netherlands. 5: 76-84. taylor & francis group, boca raton. walleyn r., vandeven é. 2006. standaardlijst van basidiomycota en myxomycota van vlaanderen en het brussels gewest. rapport inbo.r.2006.27. instituut voor natuur – en bosonderzoek, brussel, p. 1–143 + errata. wasser s.p. 1993. tribes cystodermataceae sing. and leucocoprinaceae sing. of the cis and baltic states. libri botanici 9, eching. wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland 7. w. szafer institute of botany, polish academy of sciences, kraków. wojewoda w., karasiński d. 2010. invasive macrofungi (ascomycota and basidiomycota) in poland. biological invasions in poland 1: 3-17. zelený l. 2006. taxonomic literature on the genus lepiota s. l. in the czech republic. czech mycol. 58 (3/4): 225-265. zuccherelli a., migliozzi v. 1998. prima segnalazione di leucocoprinus lilacinogranulosus v. subglobi­ sporus per il territorio italiano. boll. gr. micol. bres. 41 (3): 197-203. leucocoprinus lilacinogranulosus (henn.) locq. in poland 67 © the author(s) 2014 published by polish botanical society leucocoprinus lilacinogranulosus (henn.) locq. w polsce streszczenie w polsce stwierdzono dotychczas występowanie pięciu gatunków z rodzaju czubnik (leuco­ coprinus): l. badhamii, l. birnbaumii, l. caepistipes, l. straminellus i l. cygneus. owocniki tych grzybów obserwowane były przeważnie w pomieszczeniach zamkniętych (szklarnie, mieszkania, centra handlowe), w donicach z roślinami ozdobnymi, jednakże część z nich pojawia się poza budynkami, na siedliskach antropogenicznych i półnaturalnych. leucocoprinus lilacinogranulosus jest gatunkiem szeroko rozpowszechnionym w europie. jest uznawany za gatunek obcy dla bioty europy, jednakże jego pochodzenie nie zostało ostatecznie ustalone. nie był on dotychczas podawany z obszaru naszego kraju. w 2012 r. zebrano owocniki tego gatunku w miejscowości bystra k. bielska białej. wyrosły one w prywatnym mieszkaniu, w terrarium, na podłożu torfowym. w pracy przedstawiono opis cech makroi mikroskopowych, opracowany na podstawie znalezionych owocników. ponadto przedyskutowano zagadnienia taksonomiczne oraz rozmieszczenie l. lilacinogranulosus. 2014-06-30t22:15:29+0200 polish botanical society 2014-08-27t17:37:48+0200 polish botanical society first records of lecanora semipallida (lichenized fungi) from romania lucyna śliwa laboratory of lichenology, w. szafer institute of botany polish academy of sciences, lubicz 46, pl-31-512 kraków, l.sliwa@botany.pl śliwa l.: first records of lecanora semipallida (lichenized fungi) from romania. acta mycol. 44 (2): 173–178, 2009. lecanora semipallida is a saxicolous member of l. dispersa-group. the first localities of the species in romania come from southern carpathians (bucegi mts.) and eastern carpathians (rodnei mts.). l. semipallida is supposed to be a common lichen species in europe. a species of similar appearance is l. dispersa s.str. a key character distinguishing l. semipallida is the presence of epithecial granules that are soluble in koh. the presence of vinetorin, often causing yellowish colour of apothecia and resulting in positive spot tests and uv reactions of the apothecial disc, is also diagnostic. apart from romania, the species is also reported from austria, bulgaria, croatia, estonia, hungary, slovakia, and ukraine. key words: lecanora dispersa-group, lichen distribution, central europe, balkan peninsula introduction lecanora semipallida h. magn. belongs to the l. dispersa-group that is characterized by an endolithic or endophloeic, rarely superficial thallus, small apothecia with mostly white thalline margins, and either with xanthones or else lacking any secondary metabolites (poelt et al. 1995; fröberg 1997; laundon 2003). lecanora semipallida was shown to be the correct name for the common, widespread member of the l. dispersa complex hitherto known as l. flotoviana (auct. non spreng.). moreover, l. xanthostoma cl. roux was shown to be conspecific with l. semipallida and therefore was relegated to synonymy (śliwa 2007a). a key character distinguishing l. semipallida is the presence of epithecial granules that are soluble in k. the presence of vinetorin, often causing yellowish colour of apothecia and resulting in positive spot tests and uv reactions of the apothecial disc, is also diagnostic. morphologically, l. semipallida is a highly variable species. the size, shape and colouration of the apothecia may vary significantly. the species acta mycologica vol. 44 (2): 173–178 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 174 l. śliwa is consistent, however, in regard to anatomy (properties of epithecial granules) and chemistry (vinetorin always present). lecanora semipallida is now one of the most widely distributed saxicolous species of the group, apart from l. dispersa (pers.) sommerf. s.str. (śliwa 2007a, b). most recently it has been recognized in some extra collections available at the lichen herbarium of the w. szafer institute of botany, polish academy of sciences in kraków. the data are presented here. material and methods the study is based on recent lichen collection from romania (by karina wilk, anna and michał ronikier) that is housed at the kram-l herbarium. some additional collections from central europe and balkan peninsula available in the herbarium as well as reference material from somf and nat. hist. mus. were also examined for comparison. for anatomical investigations, free-hand sections were made with a razor blade and mounted in water. tissue measurements were made in water, and ascospore in 25% koh (k). granulation of tissues was observed in polarized light. the solubility of granules or/and crystals was tested with ca 25% koh (k) and 65% nitric acid (n). chemical examination included colour reactions, response to ultraviolet light (uv) and thin-layer chromatography (tlc). spot test reactions of thalli, apothecial margins and discs were made with koh [20–30% aqueous solution] (k), sodium hypochlorite [commercial laundry bleach] (c) and paraphenylenediamine [solution in 95% ethyl alcohol] (pd). the tlc analyses was performed in solvent system a or c (methods followed orange et al. 2001). results and discussion lecanora semipallida h. magn., lichens from central asia: 89. 1940. forms apothecia that occurr singly, or are clustered in groups, sessile, or constricted at the base to almost raised, flat when mature or flexuose, 0.4–1.3(–1.4) mm diam. apothecial disc is plane, smooth, yellow, pale greenish yellow, or yelloworange to pale brown, epruinose, or slightly pruinose. thalline margin is prominent or level with the disc, often considerably thick, smooth or rough, entire or distinctly crenate, often with bluish pigment. thallus is immersed within the substratum, and not apparent, or crustose, indistinct to clearly visible. occasionally pycnidia are produced on a surface of thr thallus. pycnidia are black, inconspicuous; conidia falcate – consistently filiform and characteristically curved, 10–17(–18) × 1 μm. amphithecium is thick, 70–170(–270) μm, corticate; algae fill the area below the cortex. the cortex is usually distinctly delimited, uniform, or slightly thicker at the the first record of lecanora semipallida 175 base than at the sides, 30–50 μm thick laterally and 50–70 μm thick at the base. it is composed of adglutinated hyphae or prosoplectenchymatous, obscured by granules occasionally interfering into the area below the cortex (prominent in polarized light, insoluble in k, soluble in n). parathecium is usually distinct, prosoplectenchymatous, 10–30 μm wide. epithecium is hyaline or shades of yellow or brown, granular (prominent in polarized light). granules are superficial and between paraphyses tips, fine to coarse, soluble in k and insoluble in n. sometimes epipsamma is present (insoluble in k, soluble in n). hymenium is hyaline, 50–90 μm high with indistinct subhymenium. hypothecium is also hyaline or distinctly yellow to orange (becoming more intense in k). it is composed of prosoplectenchyma and clear, without granules, confluent with parathecium and similar in colour, 50–160 μm high. paraphyses are simple or dichotomously branched at tips, slender or thickened, not expanded, or slightly expanded apically, usually not pigmented and free in k. asci are clavate to broadly clavate, 4–8-spored. ascospores are hyaline, simple, broadly ellipsoid 7.5–13 × 4.5–7.5 μm. apothecial margin reacts positively with some spot tests reagents (k+ yellow, c– or c+ yellow, kc+ yellow, pd–; disc k+ yellow or orange, c+ yellow or orange, pd–). the apothecia are also uv+ yellow-orange. the chemical constituent detected by tlc is vinetorin (5-chloro-3-o-methylnorlichexanthone). in romania l. semipallida was found on natural, calcium containing rocks and on concrete. in other parts of its range it has been found on calcareous rocks (limestone, lime-rich sandstone) as well as on concrete or overgrowing other lichens, e.g. aspicilia calcarea (l.) mudd, caloplaca spp., lecanora spp., physcia spp., phaeophyscia nigricans (flörke) moberg, verrucaria spp.; occasionally also on bark, bryophytes and plant debris, also on metal. lecanora semipallida is a widespread species in europe and north america. it occurs in the arctic, alpine to boreal and temperate region with main distribution in temperate region. the species was also noted from asia, australia and new zealand (śliwa 2007a, b). the most similar and likely to be confused species is l. dispersa s.str. from which l. semipallida differs both anatomically and chemically. lecanora dispersa is currently recognized as having epithecial granules that often extend into part or all of the hymenium and that are k insoluble. the presence of pannarin in l. dispersa is also detectable in most specimens (pd+ orange, detectable especially on inner side of the apothecial margins) (śliwa 2006). worth mentioning is that both taxa seem to have identical distribution pattern (śliwa 2007b). specimens examined. romania. eastern carpathians, rodnei mts.: mt. corongiş, 47o31’36”n, 24o47’39”e, vertical limestone rocks, alt. 1827 m, 30 july 2005, leg. k. wilk 3560, 3569b (kram), 47o31’35”n, 24o47’40”e, limestone outcrops, alt. 1812 m, leg. k. wilk 3582b (kram). southern carpathians, bucegi mts.: main plateau, slopes between cabana babele and muntele caraiman, 45°24’39”n, 25°28’25”e, alt. 2200 m, on calcareous rock, 26 july 2004, leg. a. ronikier & m. ronikier (kram-l 53321); at the pass şaua sugărilor, 45°25’54”n, 25°27’34”e, alt. 2400 m, on calcareous rock, 27 july 2004, leg. a. ronikier & m. ronikier (kram-l 53322). additional specimens examined. austria. wien city, schönbrun district, near railway station, on concrete, 21 july 2005, leg. k. wilk 3520b (kram). bulgaria. ne bulgaria, the town of shoumen: along the track to visoka polyana, on calcareous rock, november 1970, leg. b. zhelezova (somf 23477), limestone ground, on other lichens, march 1960, leg. b. zhelezova (somf 24433); osogovska planina, along the railroad, in the surroundings of zemen monastery, on rock, june 1952, leg. b. zhelezova (nat. hist. mus. 176 l. śliwa 2553); pirin mts., below vihren peak, on limestone, 27 july 1954, leg. b. zhelezova (nat. hist. mus. 2580, as l. flotowiana!); prov. velingrad, western rodopes mts., beglika nature reserve, 15 km ssw of batak, alt. ca 1600 m, on calcareous rock, 27 sept. 1975, leg. j. nowak (kram-l 30322). croatia. benkovac town, ne of biograd, walls of old castle, 25 july 2007, leg. l. śliwa 3880 (kram). estonia. harjumaa, lahemaa national park, kuusalu comm., hundikangrud, 59.49735°n, 25.51195°e, on limestone, 2 aug. 2008, leg. a. suija 106 (tu, kram); tartu co., tõrvandi, aia str. 50, 58.33222°n, 26.69944°e, on concrete, 18 jan. 2009, leg. i. jüriado (tu 45029, 45030, kram). hungary. siófok, sóstó, lake balaton, baross gábor utca, 46°56’22.6”n, 18°07’44.6”e, on concrete, 10 jan. 2009, leg. l. lőkös (bp, kram). slovakia. tatry mts., tichá dolina valley, concrete fence, 10 aug. 2005, leg. l. śliwa 3512 (kram). ukraine. lviv region, zolochiv district, holohory, hora in plugowo near plugovo village, xerothermic vegetation 10 km se of zolochiv, on calcareous rock, 49°44’55”n, 25°01’32”e, alt. 291 m, 23 june 2003, leg. l. śliwa 1840, 1855 (kram). conclusion lecanora semipallida it is one of the more distinct species of the group and most widespread, especially in temperete region of europe and north america. the species was reported from many countries as l. xanthostoma, e.g., clauzade and roux (1985), nimis (1993), pišút et al. (1996), llimona and hladun (2001), bielczyk et al. (2005) and lisická (2005). most likely it was also recorded under the name l. flotov[w]iana, e.g., john (1996), diederich and sérusiaux (2000), dolnik and petrenko (2003), clerc (2004) and mayrhofer et al. (2005). in the latter case, however, any renaming of specimens should be made only after their thorough and critical re-determination since the name l. flotoviana has been found to have been applied to a range of different species. full details on the status and application of the above names are provided in śliwa (2007a). a key applicable to all species of the l. dispersa complex identification is included in the recent revision of the complex in north america (śliwa 2007a). acknowledgements. dr. karina wilk, dr. anna ronikier and dr. michał ronikier (kraków) are thanked for making their collection available for study. sincere thanks are due to dr. dimitar stoykov (sofia) for the loan of specimens from somf and nat. hist. mus., as well to inga jüriado (tartu)and dr. laszlo lőkös(budapest) for providing the reference material. i am indebted to prof. dr hab. krystyna czyżewska (łódź) for comments on a paper proposal. i am also grateful to the anonymous reviewer for valuable suggestions on the manuscript. the study was supported by the ministry of science and higher education, grant no. n304 05032 2318. references bielczyk u., bylińska e., czarnota p., czyżewska k., guzow-krzemińska b., hachułka m., kiszka j., kowalewska a., krzewicka b., kukwa m., leśniański g., śliwa l., zalewska a. 2005. contribution to the knowledge of lichens and lichenicolous fungi of western ukraine. polish bot. j. 50 (1): 39–64. clauzade g., roux c. 1985. likenoj de okcidenta e�ropo. ilustrita determinlibro. bull. soc. bot. cen-likenoj de okcidenta e�ropo. ilustrita determinlibro. bull. soc. bot. cen-bull. soc. bot. centre-ouest, numero special 7. royan, france, 893 pp. clerc ph. 2004. les champignons lichenisés de suisse. cryptog. helv. 19: 1–314. diederich p., sérusiaux e. 2000. the lichens and lichenicolous fungi of belgium and luxembourg. an annotated checklist. musée national d’histoire naturelle, luxembourg, 208 pp. the first record of lecanora semipallida 177 dolnik ch., petrenko d. e. 2003. lichens of the southern curonian spit in the baltic sea. bot. zhurn. 88 (2): 41–59. fröberg l. 1997. variation in the lecanora dispersa group in south sweden. symb. bot. upsal. 32 (1): 29–34. john v. 1996. preliminary catalogue of lichenized and lichenicolous fungi of mediterranean turkey. bocconea 6: 173–216. laundon j. r. 2003. the status of lecanora zosterae in the british isles. lichenologist 35 (2): 97–100. lisická e. 2005. the lichens of the tatry mountains. veda the publishing house of the slovak academy of sciences, bratislava, 439 pp. llimona x., hladun n. l. 2001. checklist of the lichens and lichenicolous fungi of the iberian peninsula and balearic islands. bocconea 14: 5–581. mayrhofer h., denchev c. m., stoykov d. y., nikolova s. o. 2005. catalogue of the lichenized and lichenicolous fungi in bulgaria. mycologia balcanica 2 (1): 3–61. nimis p. l. 1993. the lichens of italy. an annotated catalogue. museo regionale di scienze naturali, monografie 12, torino, 897 pp. orange a., james p. w., white f. j. 2001. microchemical methods for the identification of lichens. british lichen society, london, 101 pp. pišút i., lackovičová a., lisická e. 1996. a second checklist and bibliography of slovak lichens. biologia, bratislava 51. supplement 3: 1–79. poelt j., leuckert c., roux c. 1995. die arten der lecanora dispersa-gruppe (lichenes, lecanoraceae) auf kalkreichen gesteinen im bereich der ostalpen – eine vorstudie. biblioth. lichenol. 58: 289– 333. śliwa l. 2006. the typification of lecanora dispersa and l. albescens. mycotaxon 97: 291–297. śliwa l. 2007a. lecanora semipallida, the correct name for l. xanthostoma, and a reappraisal of l. flotoviana (lecanoraceae, ascomycotina). polish bot. j. 52 (1): 71–79. śliwa l. 2007b. a revision of lecanora dispersa complex in north america. polish bot. j. 52 (1): 1–70. pierwsze notowanie lecanora semipallida (grzyby zlichenizowane) z rumunii streszczenie lecanora semipallida h. magn. należy do naskalnych porostów z grupy l. dispersa, która charakteryzuje się plechą wgłębioną w podłoże oraz drobnymi apotecjami z białym brzeżkiem plechowym. gatunki w tej grupie nie wytwarzają substancji porostowych lub wytwarzają ksantony. l. semipallida jest właściwą nazwą dla rozpowszechnionego taksonu z tej grupy dotąd znanego jako l. flotoviana (auct. non spreng.). niedawno wykazano, że l. semipallida jest identyczna z l. xanthostoma cl. roux i dlatego ten ostatni gatunek uznano za synonim (śliwa 2007a). lecanora semipallida jest obecnie gatunkiem najczęściej wyróżnianym w swojej grupie, choć w wielu krajach nie został on dotąd podany. ostatnio stwierdzono go w kilku kolekcjach, które dostępne są w zielniku kram-l. na podstawie zbiorów porostów z rumunii zebranych przez karinę wilk oraz annę i michała ronikier w niniejszej pracy l. semipallida odnotowana jest jako nowy gatunek dla tego terenu. podano również stanowiska tego gatunku z austrii, bułgarii, chorwacji, estonii, słowacji, ukrainy i węgier. kluczowe cechy wyróżniające l. semipallida to: obecność granul w epithecium, które są rozpuszczalne w k i obecność winetoryny, która powoduje żółtawy wygląd apotecjów, barwną ich reakcję na standardowe odczynniki (c+, k+ żółte lub pomarańczowe) oraz świecenie w uv. gatunek ten występuje na skałach wapiennych, jak również na cemencie i na zaprawie murarskiej. często osiedla się także na innych porostach, np. na aspicilia calcarea (l.) mudd, 178 l. śliwa caloplaca spp., lecanora spp., physcia spp., phaeophyscia nigricans (flörke) moberg i verrucaria spp. oraz wyjątkowo również na korze drzew, mszakach i szczątkach roślin, a także na metalu. l. semipallida to gatunek rozpowszechniony na świecie. występuje często w ameryce północnej i w europie; zarówno w strefie arktycznej, alpejskiej, borealnej jak i umiarkowanej. gatunek ten najczęściej jest mylony z l. dispersa s.str. taksony te różnią się jednak anatomicznie i chemicznie. l. dispersa można rozpoznać dzięki granulom w epithecium, które często przechodzą do hymenium i są nierozpuszczalne w k. u większości okazów l. dispersa można również wykryć pannarynę, która powoduje reakcję pozytywną z pd (pd+ pomarańczowe). warto podkreślić, że oba gatunki wykazują podobny wzorzec rozmieszczenia. 2014-01-01t11:49:17+0100 polish botanical society 2014-08-28t14:43:07+0200 polish botanical society 2014-08-25t14:56:58+0200 polish botanical society endangered macrofungi of selected nature reserves in wielkopolska maria lisiewska department of plant ecology and environmental protection, adam mickiewicz university umultowska 89, pl 61 614 poznań l i s i e w s k a m .: endangered macrofungi of selected nature reserves in wielkopolska. acta mycol. 41 (2): 241 252, 2006. the article presents localities of 123 species of endangered macrofungi found between 1955 and 2005 in 25 nature reserves in the wielkopolska region, where mycocoenological studies were carried out. the distribution of these reserves is shown in the cartogram based on the atpol grid. key words: macrofungi, endangered species, distribution, nature reserves, wielkopolska region introduction several countries have published the national red lists of endangered fungi, and the provisional list of 278 species of endangered european macrofungi has been prepared as well (i n g 1993). so far, two editions of the national red list of threatened macrofungi in poland (wo j e w o d a , ł a w r y n o w i c z 1992) and recently the third edition have appeared (wo j e w o d a , ł a w r y n o w i c z 2006). three regional lists have been published as well. these regional lists, based on repeated observations of macrofungi in various types of habitats, have been elaborated for the polish carpatians (wo j e w o d a 1991), upper silesia (wo j e w o d a 1999) and the góry świętokrzyskie mts. (ł u s z c z y ń s k i 2002). single data on threatened macrofungi according to the polish red list (wo j e w o d a , ł a w r y n o w i c z 1992) in the wielkopolska region may be found in some mycocoenological publications. to the date only distribution of the strictly protected by law macrofungi in the wielkopolska region has been shown in cartograms based on the atpol grid (l i s i e w s k a , m a d e j a 2003). this paper presents a list of threatened macrofungi occurring in wielkopolska, which have found their refuges in the nature reserves, where mycocoenological studies were carried out. acta mycologica vol. 41 (2): 241-252 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 242 m. lisiewska materials and methods the basic materials for this paper come from already published literature as well as unpublished mycological master’s theses made and deposited in the department of plant ecology and environmental protection at the adam mickiewicz university in poznań. into consideration were taken papers presenting results of mycocoenological studies conducted in phytocoenoses of different plant associations in 25 nature reserves of wielkopolska in the years 1955-2005. the list of macrofungi includes species names and divided after wo j e w o d a and ł a w r y n o w i c z (1992) into five categories of threat: ex – extinct, e – endangered, v – vulnerable, r – rare and i – indeterminate. the distribution of the researched nature reserves is shown in the cartogram based on the grid square method acc. to ż u k o w s k i et al. (2001) (fig. 1), adapted to fungi (wo j e w o d a 2000, 2002; l i s i e w s k a , m a d e j a 2003). large squares are of 100x100 km, smaller are of 10 x 10 km size (fig. 2). in the list of species (ordered alphabetically within each category of endangerment), after the number of the reserve (see fig. 2), the following abbreviations for scientific names of plant associations (acc. to b r z e g , wo j t e r s k a 2001) are used: a-f – astrantio-fraxinetum aa-q – aulacomnio androgyni-quercetum roboris ( carpino-quercetum) ap – abietetum albae ( a. polonicum) ca-q – calamagrostio arundinaceae-quercetum petraeae ce-a – carici elongatae-alnetum ( ribeso nigri-alnetum) cv-p – calamagrostio villosae-pinetum df-f – deschampsio flexuosae-fagetum ( fago-quercetum, luzulo pilosae-fagetum) f-a – fraxino-alnetum ( circaeo-alnetum) g-c – galio sylvatici-carpinetum l-p – leucobryo-pinetum l-q – luzulo-quercetum petraeae l-sph – ledo-sphagnetum magellanici m-f – melico uniflorae-fagetum pa-q – potentillo albae-quercetum p-c – euonymo-prunetum spinosae ( pruno spinosae-crataegetum) qr-p – querco roboris-pinetum q-u – querco-ulmetum minoris ( fraxino-ulmetum, ficario-ulmetum) sph-a – sphagno-alnetum vo-u – violo odoratae-ulmetum (=querco ulmetum minoris) vu p – vaccinio uliginosi pinetum then it follows the square number, and the author of data. endangered macrofungi 243 fig. 1. location of the study area. fig. 2. distribution of nature reserves on the atpol square grid in the wielkopolska re gion: 1. cb 17 kuźnik, 2. cb 48 źródliska flinty, 3. cb 67 bagno chlebowo, 4. cb 73 buki nad jeziorem lutomskim, 5. cb 76 świetlista dąbrowa, 6. cb 79 buczyna, 7. cb 88 gogulec, 8. cb 98 meteoryt morasko, 9. cc 50 dębina, 10. cc 91 las liściasty w promnie, 11. cc 92 bielawy, 12. db 10 uroczysko grodziszcze, 13. db 11 kręcki łęg, 14. db 18 pusz czykowskie góry, 15. pod dziadem, 16. db 28 goździk siny w grzybnie, 17. db 44 wyspa konwaliowa, 18. db 45 jezioro trzebidzkie, 19. db 92 uroczysko obiszów, 20. dc 60 czerwona róża, 21. dc 82 dąbrowa smoszew, 22. dc 96 majówka, 23. niwa, 24. eb 09 olszyny niezgodzkie, 25. ec 16 jodły ostrzeszowskie. 244 m. lisiewska list of species and their localities ex extinct serpula himantioides (fr.) bond. ex parm. – 15, g-c with pinus, db 18 (b u j a k i e w i c z , f i e b i c h 1993). e endangered cortinarius violaceus (l.: fr.) s.f. gray – 23, aa-q, dc 96 (k o z y r a 1986). geastrum melanocephalum (czern.) staněk – 12, g-c, db 10 (j u r e k 1981). geastrum rufescens pers.: pers. – 4, g-c with fagus, cb 73 (n o w a k d r ó z d 1978); 6, m-f, cb 79 (e n d l e r 1971); 8, g-c, cb 98 (c e l k a , l i s i e w s k a 1996). lentinellus omphalodes (fr.) p. karst. – 6, m-f, cb 79 (e n d l e r 1971). lycoperdon decipiens dur. et mont. – 5, pa-q, cb 76 (a n d r z e j e w s k a 2004). mycenella margaritispora (lge.) sing. – 4, f a, cb 73 (n o w a k d r ó z d 1978); 13, f a, db 11 (k o m o r o w s k a 2005). v vulnerable amanita virosa (fr.) bert. – 4, g-c with fagus, cb 73 (n o w a k d r ó z d 1978). boletus edulis bull.: fr. – 1, ca-q, cb 17 (wr ó b l e w s k a 1993); 4, g-c with fagus, cb 73 (n o w a k d r ó z d 1978); 6, m-f, cb 79 (e n d l e r 1971); 8, g-c, l-p, cb 98 (c e l k a , l i s i e w s k a 1996); 10, g-c, cc 91 (l i s i e w s k a 1961); 20, g-c with larix, dc 60 (n o w a c k a 1978); 21, g-c, dc 82 (m o l s k a 1982); 22, caq, dc 96 (k o z y r a 1986); 23, aa-q, g-c, dc 96 (k o z y r a 1986); 25, ap, ec 16 (k u c h a r s k a 1987). coprinus picaceus (bull.: fr.) s.f. gray – 4, g-c with fagus, cb 73 (n o w a k d r ó z d 1978); 6, m-f, cb 79 (e n d l e r 1971); 9, g-c, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976; l i s i e w s k a , p o ł c z y ń s k a 1998). fistulina hepatica (schaeff.) fr. – 5, pa-q, cb 76 (a n d r z e j e w s k a 2004); 9, g-c, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976; l i s i e w s k a , p o ł c z y ń s k a 1998); 12, g-c, db 10 (j u r e k 1981); 21, g-c, dc 82 (m o l s k a 1982). gyromitra gigas (krbh.) cke. – 8, g-c, cb 98 (c e l k a , l i s i e w s k a 1996). hebeloma pumilum lge. – 2, f-a, cb 48 (wa s i e l e w s k a 1999). hericium clathroides (pallas: fr.) pers. [ h. ramosum (bull.) letell] – 4, g-c with fagus, cb 73 (n o w a k d r ó z d 1978); 18, df-f, db 45 (b r z e g et al. 2005). hygrocybe cantharellus (schw.) murrill – 2, f-a, cb 48 (wa s i e l e w s k a 1999). hypholoma elongatum (pers.: fr.) rick. – 7, sph-a, cb 88 (p a w l a k 2003). lactarius sanguifluus fr. – 8, ca-q, cb 98 (c e l k a , l i s i e w s k a 1996). morchella esculenta (l.) pers. – 4, g-c with fagus cb 73 (n o w a k d r ó z d 1978); 21, g-c, dc 82 (m o l s k a 1982); 22 ca-q, dc 96 (k o z y r a 1986). mycena megaspora kauffm. – 7, sph-a, cb 88 (p a w l a k 2003). mycena olida bres. – 14, vo-u, a-f, db 18 (s u s i c k a 1994; w i l c z y ń s k a 1994). endangered macrofungi 245 mycena pullata (berk. et cke.) sacc. – 4, f-a, cb 73 (n o w a k d r ó z d 1978); 14, q-u, vo-u, db 18 (s u s i c k a 1994). oudemansiella mucida (schrad.: fr.) höhn. – 4, g-c with fagus, cb 73 (n o w a k d r ó z d 1978); 6, m-f, cb 79 (e n d l e r 1971); 25, ap, ec 16 (k u c h a r s k a 1987). pleurotus cornucopiae (paul.: pers.) rolland – 14, q-u, vo-u, db 18 (s u s i c k a 1994). psathyrella caput-medusae (fr.) konr. et maubl. – 17, f-a, q-u, db 44 (k r z y ż a n o w s k a 1977). trametes pubescens (schum.: fr.) pil. – 25, ap, ec 16 (k u c h a r s k a 1987). tricholoma populinum lge. – 12, g-c, db 10 (j u r e k 1981). xerula pudens (pers.) sing. [ oudemansiella longipes (quél.) mos.] – 5, pa-q, cb 76 (a n d r z e j e w s k a 2004); 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998); 19, g-c, db 92 (l i s i e w s k a , s e k u ł a -wo ź n i a k 1998). r rare agaricus xanthodermus gen. – 14, q-u, db 18 (s u s i c k a 1994). amanita aspera (fr.) s.f. gray – 22, ca-q, dc 96 (k o z y r a 1986). antrodiella hoehnelii (bres. ex höhn.) niemelä – 2, f-a, cb 48 (wa s i e l e w s k a 1999); 7, sph-a, cb 88 (p a w l a k 2003); 14, q-u, db 18 (s u s i c k a 1994); 24, ce-a, eb 09 (b u j a k i e w i c z 1999). auricularia mesenterica (dicks.: fr.) pers. – 16, qr-p, db 28 (l i s i e w s k a , f i l i s i e w i c z 2006). bulgaria inquinans (pers.: fr.) fr. – 5, pa-q, cb 76 (a n d r z e j e w s k a 2004). ceriporia excelsa (lund.) parm. – 14, a-f, db 18 (w i l c z y ń s k a 1994). clitocybe angustissima (lasch: fr.) kumm. ss. lge. – 1, l-p, cb 17 (wr ó b l e w s k a 1993). cordyceps capitata (holmsk.: fr.) link – 1, ca-q, cb 17 (wr ó b l e w s k a 1993). cordyceps ophioglossoides (ehrenb.: fr.) link – 1, ca-q, cb 17 (wr ó b l e w s k a 1993). cortinarius acutus (pers.: fr.) fr. – 19, l-q, db 92 (l i s i e w s k a , s e k u ł a -wo ź n i a k 1998). cystoderma cinnabarinum (alb. & schw.) fay. – 6, m-f, cb 79 (e n d l e r 1971). cystoderma granulosum (batsch: fr.) fay. – 1, ca-q, cb 17 (wr ó b l e w s k a 1993). ganoderma lucidum (curt.: fr.) p. karst. – 9, g-c, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976; l i s i e w s k a , p o ł c z y ń s k a 1998); 12, g-c, db 10 (j u r e k 1981). gyrodon lividus (bull.: fr.) sacc. – 14, a-f, db 18 (w i l c z y ń s k a 1994). hypholoma marginatum (pers.) schroet. – 6, m-f, cb 79 (e n d l e r 1971). hypholoma udum (pers.: fr.) kühn. – 7, sph-a, cb 88 (p a w l a k 2003). inocybe calospora quél. – 6, ca-q, cb 79 (e n d l e r 1971). lactarius chrysorrheus fr. – 8, ca-q, cb 98 (c e l k a , l i s i e w s k a 1996). macrotyphula filiformis (bull.: fr.) paechnatz [ clavariadelphus junceus (alb. et schwein.: fr.) corner] – 7, sph-a, cb (p a w l a k 2003); 14, a-f, db 18 (w i l c z y ń s k a 1994). 246 m. lisiewska macrotyphula fistulosa (holmsk.: fr.) r.h. petersen [ clavariadelphus fistulosus (holmsk.: fr.) corner] – 2, f-a, cb 48 ( wa s i e l e w s k a 1999); 7, sph-a, cb 88 (p a w l a k 2003); 8, ca-q, cb 98 (c e l k a , l i s i e w s k a 1996); 21, g-c, dc 82 (m o l s k a 1982). melanophyllum haematospermum (bull.: fr.) kreisel [ m. echinatum (roth: fr.) sing.] – 4, f-a, cb 73 (n o w a k d r ó z d 1978). micromphale foetidum (sow.: fr.) sing. – 4, f-a, cb 73 (n o w a k d r ó z d 1978). mutinus caninus (huds.: pers.) fr. – 19, g-c, db 92 (l i s i e w s k a , s e k u ł a -wo ź n i a k 1998). mycena adonis (bull.: fr.) s.f. gray – 3, vu-p, l-sph, cb 67 (f i k l e w i c z s o b s t y l 1965); 20, g-c with larix, dc 60 (n o w a c k a 1978). mycena adscendens (lasch) maas – 16, qr-p, db 28 (l i s i e w s k a , f i l i s i e w i c z 2006). mycena flavescens vel. – 6, m-f, cb 79 (e n d l e r 1971). mycena supina (fr.) kumm. – 7, sph-a, cb 88 (p a w l a k 2003); 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998); 14, q-u, db 18 (s u s i c k a 1994). mycoacia uda (fr.) donk – 14, q-u, db 18 (s u s i c k a 1994). paxillus filamentosus (scop.) fr. – 4, f-a, cb 73 (n o w a k d r ó z d 1978); 7, sph-a, cb 88 (p a w l a k 2003). phaeolus schweinitzii (fr.) pat. – 11, g-c with larix, cc 92 (s z c z u d ł o 1979); 20, g-c with larix, dc 60 (n o w a c k a 1978); 22, ca-q, dc 96 (k o z y r a 1986). phaeomarasmius erinaceus (fr.) kühn. – 2, f-a, cb 48 (wa s i e l e w s k a 1999). phellinus ferruginosus (schrad.: fr.) pat. – 14, q-u, vo-u, db 18 (s u s i c k a 1994). phellinus pini (brot.: fr.) ames – 16, qr-p, db 28 (l i s i e w s k a , f i l i s i e w i c z 2006); 20, g-c with larix, pinus, dc 60 (n o w a c k a 1978). pholiotina filaris (fr.) sing. – 9, f-a, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976); 12, g-c, db 10 (j u r e k 1981). pisolithus arhizus (scop.: pers.) rauschert – 8, ca-q, cb 98 (c e l k a , l i s i e w s k a 1996). polyporus melanopus (pers.) fr. – 6, m-f, cb 79 (e n d l e r 1971). psathyrella albidula (romagn.) mos. – 9, q-u, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976). pycnoporus cinnabarinus (jacq.: fr.) p. karst. – 2, ce-a, f-a, cb 48 (wa s i e l e w s k a 1999). ramaria aurea (schaeff. fr.) quél – 23, g-c, dc 96 (k o z y r a 1986). ramaria botrytis (pers.: fr.) ricken – 6, m-f, cb 79 (e n d l e r 1971); 23, aa-q, g-c, dc 96 (k o z y r a 1986). ramaria formosa (pers.: fr.) quél – 22, ca-q, de 96 (k o z y r a 1986); 23, aa q, gc, dc 96 (k o z y r a 1986). russula olivacea (schaeff.) fr. – 8, g-c, cb 98 (c e l k a , l i s i e w s k a 1996). sparassis crispa (wulf.) fr. – 8, ca-q, cb 98 (c e l k a , l i s i e w s k a 1996), 16, df-f, db 28 (l i s i e w s k a , f i l i s i e w i c z 2006); 18, l-p, db 45 (b r z e g et al. 2005); 20, g-c with larix, pinus, dc 60 (n o w a c k a 1978); 22, ca-q, dc 96 (k o z y r a 1986). stereum subtomentosum pouz. – 2, ce-a, f-a, cb 48 (wa s i e l e w s k a 1999); 4, f-a, cb 73 (n o w a k d r ó z d 1978); 8, ca-q, cb 98 (c e l k a , l i s i e w s k a 1996); 9, endangered macrofungi 247 g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998); 23, aa-q, dc 96 (k o z y r a 1986); 24, ce-a, eb 09 (b u j a k i e w i c z 1999). tephrocybe rancida (fr.) mos. – 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998). thelephora caryophyllea (schaeff.) fr. – 16, df-f, db 28 (l i s i e w s k a , f i l i s i e w i c z 2006). i indeterminate agrocybe paludosa (lge.) kühn. et romagn. – 14, a-f, db 18 (w i l c z y ń s k a 1994). calyptella capula (holmsk.: fr.) quél. – 4, f-a, cb 73 (n o w a k d r ó z d 1978); 24, ce-a, eb 09 (b u j a k i e w i c z 1999). cantharellus cibarius fr. – 1, pa-q, ca-q, cb 17 (wr ó b l e w s k a 1993); 5, pa-q, cb 76 (a n d r z e j e w s k a 2004); 6, m-f, ca-q, cb 79 (e n d l e r 1971); 8, ca-q, cb 98 (c e l k a , l i s i e w s k a 1996); 10, g-c, cc 91 (l i s i e w s k a 1961); 20, g-c with larix, pinus, dc 60 (n o w a c k a 1978); 21, g-c, dc 82 (m o l s k a 1982); 22, caq, dc 96 (k o z y r a 1986); 23, g-c, aa-q, dc 96 (k o z y r a 1986); 25, cv-p, ec 16 (k u c h a r s k a 1987). clitocybe candicans (pers.: fr.) kühn. – 4, f-a, cb 73 (n o w a k d r ó z d 1978); 6, mf, cb 79 (e n d l e r 1971); 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998); 19, l-q, db 92 (l i s i e w s k a , s e k u ł a -wo ź n i a k 1998); 25, ap, ec 16 (k u c h a r s k a 1987). collybia putilla (fr.) sing. [ gymnopus putillus (fr.: fr.) antonin, halling & noordel.] – 16, l-p, db 28 (l i s i e w s k a , f i l i s i e w i c z 2006). cordyceps militaris (l.: fr.) link – 2, ce-a, f-a, cb 48 (wa s i e l e w s k a 1999); 11, g-c with larix, cc 92 (s z c z u d ł o 1979); 21, g-c, dc 82 (m o l s k a 1982). cortinarius incisus (pers.: fr.) fr. – 1, ca-q, l-p, cb 17 (wr ó b l e w s k a 1993). cortinarius orellanus fr. – 11, g-c with larix, cc 92 (s z c z u d ł o 1979); 23, aa-q, dc 96 (k o z y r a 1986). cortinarius rigidus (scop.) fr. – 1, ca-q, pa-q, cb 17 (wr ó b l e w s k a 1993); 11, g-c with larix, cc 92 (s z c z u d ł o 1979). cortinarius rubricosus (fr.) fr. – 5, pa-q, cb 76 (a n d r z e j e w s k a 2004). cortinarius saniosus (fr.) fr. – 9, f-a, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976). cortinarius triumphans fr. – 1, pa-q, cb 17 (wr ó b l e w s k a 1993). cortinarius uraceus fr. – 8, ca-q, cb 98 (c e l k a , l i s i e w s k a 1996). datronia mollis (sommerf.: fr.) donk – 14, q-u, a-f, db 18 (s u s i c k a 1994; w i l c z y ń s k a 1994). entoloma asprellum (fr.) mos. – 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998). entoloma juncinum (kühn. et romagn.) noordel. [ rhodophyllus junceus ss. lge.] – 5, pa-q, cb 76 (a n d r z e j e w s k a 2004); 6, m-f, cb 76 (e n d l e r 1971); 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998); 11, g-c, cc 92 (s z c z u d ł o 1979); 16, df-f, db 28 (l i s i e w s k a , f i l i s i e w i c z 2006); 19, g-c, db 92 (l i s i e w s k a , s e k u ł a -wo ź n i a k 1998); 21, g-c, dc 82 (m o l s k a 1982). entoloma prunuloides (fr.: fr.) quél. – 23, g-c, dc 96 (k o z y r a 1986). entoloma rhodocylix (lasch: fr.) mos. – 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998). entoloma rusticoides (gill.) noordel. – 7, sph-a, cb 88 (p a w l a k 2003). 248 m. lisiewska galerina badipes (fr.) kühn. – 3, vu-p, cb 67 (f i k l e w i c z s o b s t y l 1965). galerina mycenoides (fr.) kühn. – 23, g-c, dc 96 (k o z y r a 1986). galerina paludosa (fr.) kühn. – 3, l-sph, cb 67 (f i k l e w i c z s o b s t y l 1965); 7, sph-a, cb 88 (p a w l a k 2003); 25, cv-p, ec 16 (k u c h a r s k a 1987). galerina sphagnorum (pers.: fr.) kühn. – 3, l-sph, cb 67 (f i k l e w i c z s o b s t y l 1965); 7, sph-a, cb 88 (p a w l a k 2003). galerina triscopa (fr.) kühn. – 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998); 14, a-f, db 18 (w i l c z y ń s k a 1994); 17, ca-q, db 44 (k r z y ż a n o w s k a 1977). gyroporus castaneus (bull.: fr.) quél. – 5, pa-q, cb 76 (a n d r z e j e w s k a 2004); 9, g-c, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976). lactarius lilacinus (lasch: fr.) fr. – 2, ce-a, f-a, cb 48 (wa s i e l e w s k a 1999); 8, q-u, cb 98 (c e l k a , l i s i e w s k a 1996). lactarius resimus (fr.) fr. – 17, ca-q, db 44 (k r z y ż a n o w s k a 1977). lepiota setulosa lge. – 9, q-u, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976; l i s i e w s k a , p o ł c z y ń s k a 1998). lactarius sphagneti (fr. in lindbl.) neuh. ex gröger – 25, ap, ec 16 (k u c h a r s k a 1987). lepiota tomentella lge. – 6, m-f, cb 76 (e n d l e r 1971); 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998). lycoperdon echinatum pers.: pers. – 10, g-c, cc 91 (l i s i e w s k a 1961). macrolepiota procera (scop.: fr.) sing. – 4, g-c, cb 73 (n o w a k d r ó z d 1978); 5, pa-q, cb 76 (a n d r z e j e w s k a 2004); 6, m-f, cb 79 (e n d l e r 1971); 8, caq, g-c, cb 98 (c e l k a , l i s i e w s k a 1996); 9, g-c, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976; l i s i e w s k a , p o ł c z y ń s k a 1998); 11, g-c, cc 92 (s z c z u d ł o 1979); 17, ca-q, g-c, db 44 (k r z y ż a n o w s k a 1977); 19, l-q, g-c, db 92 (l i s i e w s k a , s e k u ł a -wo ź n i a k 1998); 20, g-c with larix, dc 60 (n o w a c k a 1978); 21, g-c, dc 82 (m o l s k a 1982); 25, ap, ec 16 (k u c h a r s k a 1987). macrolepiota rhacodes (vitt.) sing. – 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998); 10, g-c, cc 91 (l i s i e w s k a 1961); 14, q-u, vo-u, db 18 (s u s i c k a 1994); 17, ca-q, f-a, g-c, db 44 (k r z y ż a n o w s k a 1977); 19, l-q, g-c, db 92 (l i s i e w s k a , s e k u ł a -wo ź n i a k 1998); 22, ca-q, dc 96 (k o z y r a 1986); 23, g-c, dc 96 (k o z y r a 1986). marasmius recubans quél. – 4, f-a, cb 73 (n o w a k d r ó z d 1978); 14, a-f, db 18 (w i l c z y ń s k a 1994); 21, g-c, d c 82 (m o l s k a 1982). mycena concolor (lge.) a.h. smith – 3, l-sph, cb 67 (f i k l e w i c z s o b s t y l 1965). mycena pelianthina (fr.) quél. – 9, g-c, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976; l i s i e w s k a , p o ł c z y ń s k a 1998); 21, g-c, dc 82 (m o l s k a 1982). mycena pterigena (fr.: fr.) kumm. – 2, f-a, cb 48 (wa s i e l e w s k a 1999). peniophora limitata (chaik.: fr.) cke. – 14, q-u, db 18 (s u s i c k a 1994). phaeomarasmius carpophilus (fr.) sing. – 4, g-c with fagus, cb 73 (n o w a k d r ó z d 1978). pluteus pseudoroberti mos. et stangl – 4, f-a, cb 73 (n o w a k d r ó z d 1978); 9, g-c, cc 50 (l i s i e w s k a , b u j a k i e w i c z 1976); 21, g-c, dc 82 (m o l s k a 1982). psathyrella frustulenta (fr.) a.h. smith – 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998). endangered macrofungi 249 psathyrella nolitangere (fr.) pears. et dennis – 1, ca-q, cb 17 (wr ó b l e w s k a 1993); 4, f-a, cb 73 (n o w a k d r ó z d 1978); 9, g-c, cc 50 (l i s i e w s k a , p o ł c z y ń s k a 1998); 14, a-f, db 18 (w i l c z y ń s k a 1994). russula alutacea (pers.: fr.) fr. – 3, vu-p, cb 67 (f i k l e w i c z s o b s t y l 1965); 6, ca-q, cb 79 (e n d l e r 1971); 19, l-q, g-c, db 92 (l i s i e w s k a , s e k u ł a wo ź n i a k 1998); 23, aa-q, dc 96 (k o z y r a 1986). russula curtipes moell. & j. schff. – 6, m-f, cb 79 (e n d l e r 1971). stropharia albocyanea (desm.) quél. [ psilocybe caerulea (kreisel) noordel] – 14, vo-u, db 18 (s u s i c k a 1994); 16, l-p, db 28 (l i s i e w s k a , f i l i s i e w i c z 2006); 17, g-c, db 44 (k r z y ż a n o w s k a 1977). stropharia squarrosa (pers.: fr.) quél. – 6, m-f, cb 79 (e n d l e r 1971); 20, g-c with larix, dc 60 (n o w a c k a 1978). tephrocybe palustris (peck) donk – 7, sph-a, cb 88 (p a w l a k 2003); 25, ap, cv-p, ec 16 (k u c h a r s k a 1987). volvariella bombycina (schaeff.: fr.) sing. – 19, l-q, db 92 (l i s i e w s k a , s e k u ł a wo ź n i a k 1998). volvariella murinella (quél.) mos. – 4, g-c with fagus, cb 73 (n o w a k d r ó z d 1978); 12, g-c, db 10 (j u r e k 1981). xerocomus armeniacus (bull.: fr.) quél. – 5, pa-q, cb 76 (a n d r z e j e w s k a 2004). results and discussion as the result of the mycocoenological studies carried out in 25 nature reserves of wielkopolska 123 species of threatened macrofungi there were recorded. from this list only one species – serpula himantioides, quoted as an extinct species in the polish red list of macrofungi, was fairly abundant in the reserve “pod dziadem” in the wielkopolski national park (b u j a k i e w i c z , f i e b i c h 1993). six species are considered to be endangered with extinction (category e), among them two protected species of geastrum (g. melanocephalum and g. rufescens). to the category v – vulnerable belong 20 species. some of them are rather common in wielkopolska, e.g. boletus edulis was found in 10 reserves, mainly in the oak forest communities. three other species which are under strict protection are worthy of notice, namely hericium clathroides, morchella esculenta and fistulina hepatica. several localities of morchella esculenta and fistulina hepatica were already recorded in the wielkopolska region, but only one locality of hericium clathroides in the reserve “buki nad jeziorem lutomskim” was shown on the published cartogram (l i s i e w s k a , m a d e j a 2003). recently, a new locality of this interesting species, connected with fagus, was found in the acidophilous beech forest association deschampsio flexuosae-fagetum in the reserve “jezioro trzebidzkie” (b r z e g et al. 2005). a large number namely 46 rare species (the category r), were reported from the researched reserves, among them also protected sparassis crispa, ganoderma lucidum and mutinus caninus (dz. u. nr 168). the category of indeterminated threat (i) was the richest in species of macrofungi in the study area. among 50 noted species there are common fungi occurring numerously in wielkopolska, e.g. cantharellus cibarius, macrolepiota procera and 250 m. lisiewska m. rhacodes. these are excluded in new edition of the red list in poland (wo j e w o d a , ł a w r y n o w i c z 2006). the investigated nature reserves are located mostly in the central part of wielkopolska (fig. 2), where the majority of mycocoenological studies were carried out. as far as only in 25 nature reserves (on the whole number of about 70 floristic and forest reserves in wielkopolska) mycological observations were conducted, the presented distribution of endangered macrofungi in protected areas of wielkopolska is certainly not comprehensive and further mycocoenological investigations in the whole region should be continued. references a n d r z e j e w s k a d. 2004. udział macromycetes w zbiorowiskach leśnych rezerwatu „świetlista dąbro wa” koło obrzycka (wielkopolska). praca magisterska z zakł. ekol. rośl. i ochr. środ. uam. po znań (msc.). b r z e g a., g ą b k a m., n a g e n g a s t b. 2005. szata roślinna rezerwatu przyrody „jezioro trzebidzkie”. biul. park. krajobraz. wielkopolski 11(13): 153 196. poznań. b r z e g a., wo j t e r s k a m. 2001. zespoły roślinne wielkopolski, ich stan poznania i zagrożenie. 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(poland). polish bot. j. 47 (2): 183 193. m o l s k a s . 1982. badania nad udziałem macromycetes w grądach rezerwatu „dąbrowa smoszew” w nadleśnictwie krotoszyn. praca magisterska z zakł. ekol. rośl. i ochr. środ. uam. poznań (msc.). n o w a c k a k . 1978. grzyby wyższe rezerwatu modrzewiowego „czerwona róża” (woj. leszczyńskie). praca magisterska z zakł. ekol. rośl. i ochr. środ. uam. poznań (msc.). n o w a k d r ó z d e . 1978. udział grzybów wyższych w płatach łęgu olszowego circaeo alnetum w rezer wacie „buki lutomskie” koło sierakowa. praca magisterska z zakł. ekol. rośl. i ochr. środ. uam. poznań (msc.). p a w l a k m . 2003. udział grzybów w fitocenozach olsu torfowcowego sphagno squarrosi alnetum w re zerwacie gogulec koło poznania. praca magisterska z zakł. ekol. rośl. i ochr. środ. uam. poznań (msc.). s u s i c k a m. 1994. udział grzybów (macromycetes) w fitocenozach łęgu violo odoratae ulmetum (we evers 1940) doing 1962 i łęgu wiązowego ficario ulmetum campestris (matuszkiewicz 1982) w rezer wacie puszczykowskie góry w wielkopolskim parku narodowym. praca magisterska z zakł. ekol. rośl. i ochr. środ. uam. poznań (msc.). s z c z u d ł o k . 1979. badania nad udziałem macromycetes w grądach rezerwatu bielawy w nadleśnictwie gniezno. praca magisterska z zakł. ekol. rośl. i ochr. środ. uam. poznań (msc.). wa s i e l e w s k a e. 1999. udział grup ekologicznych grzybów (macromycetes) w fitocenozach olsu po rzeczkowego (ribo nigri alnetum) i łęgu olszowego (circaeo alnetum) na terenie rezerwatu „źródli ska flinty” w nadleśnictwie sarbia (województwo wielkopolskie). praca magisterska z zakł. ekol. rośl. i ochr. środ. uam. poznań (msc.). w i l c z y ń s k a i. 1994. udział grzybów w fitocenozie łęgu astrantio fraxinetum w rezerwacie puszczykow skie góry w wielkopolskim parku narodowym. praca magisterska z zakł. ekol. rośl. i ochr. środ. uam. poznań (msc.). wo j e w o d a w. 1991. pierwsza czerwona lista grzybów wielkoowocnikowych (macomycetes) zagrożo nych w polskich karpatach. first red list of threatened macrofungi in the polish carpathians. stud. ośrod. dokument. fizjogr. pan, 18: 239 261. kraków. wo j e w o d a w. 1999. czerwona lista grzybów wielkoowocnikowych górnego śląska. red list of up per silesian macrofungi. centrum dziedzictwa przyrody górnego śląska. raporty, opinie 4: 8 51. katowice. wo j e w o d a w. (ed.). 2000. atlas of the geographical distribution of fungi in poland, fasc. 1, pp. 61. w. szafer inst. bot. pol. acad. sci., kraków. wo j e w o d a w. (ed.). 2002. atlas of the geographical distribution of fungi in poland, fasc. 2, pp. 129. w. szafer inst. bot. pol. acad. sci., kraków. wo j e w o d a w., ł a w r y n o w i c z m. 1992. red list of threatened macrofungi in poland. (in:) k. z a r z y c k i , w. wo j e w o d a , z. h e i n r i c h (eds). list of threatened plants in poland. 2 ed.: 27 56. w. szafer inst. bot. polish acad. sci., kraków. wo j e w o d a w., ł a w r y n o w i c z m. 2006. red list of the macrofungi in poland. (in:) z. m i r e k , k. z a r z y c k i , w. wo j e w o d a , z. s z e l ą g (eds). red list of plants and fungi in poland: 53 70. w. szafer inst. bot. polish acad. sci., kraków. wr ó b l e w s k a k. 1993. macromycetes wybranych zbiorowisk leśnych rezerwatu „kuźnik” koło piły. pra ca magisterska z zakł. ekol. rośl. i ochr. środ. uam. poznań (msc.). ż u k o w s k i w., c e l k a z., c h m i e l j., j a c k o w i a k b., l a t o w s k i k., s z k u d l a r z p. 2001. rozmiesz czenie wybranych gatunków roślin ginących w wielkopolsce. prace zakł. taksonomii roślin uam w poznaniu, 12. ss. 68. bogucki wyd. nauk., poznań. 252 m. lisiewska zagrożone macromycetes wybranych rezerwatów przyrody w wielkopolsce s t r e s z c z e n i e praca zawiera stanowiska 123 gatunków zagrożonych grzybów wielkoowocnikowych, zna lezionych w wielkopolsce na terenie 25 rezerwatów przyrody, w których prowadzone były badania mikosocjologiczne w latach 1955 2005. rozmieszczenie tych rezerwatów przed stawiono na kartogramie w siatce kwadratów atpol. jako jedno stanowisko przyjmuje się wszystkie notowania w obrębie jednego małego kwadratu (10x10 km). wykaz gatunków grzybów uporządkowano według kategorii zagrożenia (wo j e w o d a , ł a w r y n o w i c z 1992). przy każdym gatunku podano numer rezerwatu (ryc. 2), skrót nazwy zespołu roślinnego, w którego fitocenozie był notowany, jak również numer kwadratu i nazwi sko autora. wśród odnotowanych w rezerwatach gatunków grzybów serpula himantioides została zaliczona do kategorii ex wymarłych i zaginionych. znaleziono 6 gatunków z kategorii e wymierających, a wśród nich objęte ścisłą ochroną geastrum melanocephalum i g. rufe scens. do kategorii v narażonych należy 20 gatunków. niektóre z nich są dość pospolite w wielkopolsce, jak np. boletus edulis podawany z 10 badanych rezerwatów. godne uwagi są ga tunki objęte ścisłą ochroną, jak hericium clathroides, morchella esculenta i fistulina hepatica. znaleziono znaczną liczbę 46 gatunków rzadkich (kategoria r), w tym również ściśle chro nione: sparassis crispa, ganoderma lucidum i mutinus caninus (dz. u. nr 168). najbogatsza była kategoria i, zawierająca gatunki macromycetes o nieokreślonym zagrożeniu. spośród 50, z tej kategorii, gatunków niektóre należą do często spotykanych na badanym terenie, jak np. cantharellus cibarius, macrolepiota procera, czy m. rhacodes. niniejsza praca może stanowić uzupełnienie wiedzy o występowaniu zagrożonych gatun ków macromycetes na obszarze wielkopolski z punktu widzenia ochrony bioróżnorodności gatunków. 2014-01-01t11:44:28+0100 polish botanical society 2014-09-06t20:24:41+0200 polish botanical society 2014-08-28t14:43:56+0200 polish botanical society preliminary studies of fungi in the biebrza national park (ne poland). i. micromycetes małgorzata ruszkiewicz-michalska1, cezary tkaczuk2, maria dynowska3, ewa sucharzewska3, jarosław szkodzik4 and marta wrzosek5 1department of mycology, faculty of biology and environmental protection university of łódź, banacha 12/16, pl-90-237 łódź, mrusz@biol.uni.lodz.pl 2department of plant protection, siedlce university of natural sciences and humanities prusa 14, pl08-110 siedlce, tkaczuk@uph.edu.pl 3department of mycology, warmia and mazury university in olsztyn oczapowskiego 1a, pl-10-957 olsztyn, dynow@uwm.edu.pl, ewko@uwm.edu.pl 4nature & ecology of łódź macroregion website, ekolodzkie.pl, jaroslaw.szkodzik@ekolodzkie.pl 5department of plants systematics and geography, university of warsaw al. ujazdowskie 4, pl-00-478 warszawa, mwrzosek@biol.uw.edu.pl ruszkiewicz-michalska m., tkaczuk c., dynowska m., sucharzewska e., szkodzik j., wrzosek m.: preliminary studies of fungi in the biebrza national park (ne poland). i. micromycetes. acta mycol. 47 (2): 213–234, 2012. this paper presents the results of the first short-term inventory of fungi species occurring in the biebrza national park, one of the biggest and best preserved protected areas of poland. the paper is focused on a survey of microfungi. fungi were collected in early autumn 2012, within the framework of a scientific project by the polish mycological society. the results are published in two parts containing microand macromycetes, respectively. an annotated list of 188 identified taxa covers true fungi including 33 zygomycetes, 130 ascomycetes (including anamorphs) and 22 basidiomycetes, as well as two chromistan and one protozoan fungal analogues. the identified fungi taxa, inhabiting diverse ecological niches, represent a wide range of trophic groups including biotrofic and necrotrophic parasites of plants, pathogens of arthropods, fungicolous fungi and saprotrophs isolated from soil and organic matter. from 188 annotated taxa, 89% (167 species) have not been recorded in the biebrza national park until now and four species are newly reported for poland (alternaria nobilis, clonostachys solani, mariannaea elegans, metasphaeria cumana). data on the species richness and taxonomic diversity of the identified fungi are briefly commented in terms of micromycetes role in managing nature conservation. key words: all-species inventory, micromycetes, plant parasites, arthropod pathogens, soil fungi, fungal ecology, protected area acta mycologica vol. 47 (2): 213–234 2012 214 m. ruszkiewicz-michalska et al. introduction a comprehensive knowledge of the organisms inhabiting national parks provides a basis for developing and improving species protection rules and range. for many hemerophobic species, national parks are a kind of lair where populations of rare species can survive and thrive and spread to adjacent areas. in general, reconnoitring of the biota of organisms occurring in a particular area is classically the first, necessary step in planning and processing their protection. this concerns fungi in particular as they are symbionts (mutualists and antagonists) and decomposers that crucially influence ecosystem functioning. fungi, often neglected in the past, are currently more often considered in conservation plans (dahlberg et al. 2011), however, it is still necessary to understand and enlighten the role of micromycetes in nature conservation strategies (e.g., denchev 2005; ingram 2002). as many micromycetes are parasites and pathogens, they are usually considered as unworthy of protection and are very rarely included in the red lists (e.g., helfer 1993; denchev 2005; minter 2007, 2011). the biebrza national park (bbnp) is relatively young (established in 1993) and is the largest (59.223ha, with a 66.824ha buffer zone) national park in poland. the park was brought to life to conserve the entire river valley, from its sources to the mouth, making it unique within europe (dyrcz, werpachowski 2005). the natural longitudinal and transversal zonation of habitats and corresponding plant communities of the river valley, as well as a large complex of fens are preserved in an almost unchanged state. peatlands are predominant among the habitats of the biebrza valley, occupying approximately 82.000ha, of which two thirds are within the area of the national park. forest communities (mainly alder and birch swamp forests) occupy only ca 26% of the bbnp area. to the total habitat and plant diversity of the bbnp greatly contribute diverse mineral ‘islands’ (called grądzik or grąd). they are characterized by high heterogenity of habitats and related plant communities (aquatic, semi-aquatic, rushes, thickets, forests, grasslands and anthropogenic ones) (werpachowski 2005). although these ‘islands’ constitute only 1% of the total area of the biebrza valley, they host about 80% of its total flora (appraised on 1015 plant species). the only group of fungi studied in relative depth in the area of the bbnp are soil mitosporic zygomycetes and ascomycetes (tyszkiewicz 2004a, b, 2012). however, works by tyszkiewicz (l.c.) present the variability of the fungi in peat profiles, rather than their diversity in different plant communities. particular attention has been paid to soiland litter-inhabiting micromycetes (mainly zygomycetes and anamorphic fungi) from biele suchowolskie fen after a severe, long-lasting fire which happened in 2002 (wilk 2009; boulahdjel 2010; budziszewska et al. 2010). in the survey by budziszewska et al. (2010), 41 zygomycetous species of soil fungi are listed from the area of the biebrza valley. the authors compared burnt and unburnt areas in terms of fungal richness and diversity. no inventory research on other groups of non-lichenized fungi has been conducted in the bbnp while the lichens have been surveyed by kolanko (2005, incl. literature cited therein). creating an inventory of fungi and slime mould biodiversity in the vast and diverse area of the bbnp is the first scientific project of the polish mycological society, micromycetes in the biebrza national park 215 officially created at the end of 2011. the results of the short-term studies on fungi are presented in two consecutive papers. this one deals with species classified as micromycetes, while the second part (kujawa et al. 2012, this issue) covers macromycetes. a few other micromycetes new to poland will be the subject of forthcoming papers. materials and methods the study was carried out on 28.08.–01.09.2012 in the central area of the bbnp and in particular in three protective units (grzędy, kapice and werykle). some observations were also conducted at the area of the osowiec fortress, located close to the park border. a map of the surveyed localities is included in the paper by kujawa et al. (2012). materials consisted of the fungi of different taxonomic and trophic groups, inhabiting diverse substrates and ecological niches, arbitrarily treated in the paper as micromycetes (in the way adopted in polish checklists, cf. mułenko et al. 2008, wojewoda 2003 and chmiel 2006). fungi were collected according to group-specific methods (cf. mułenko 2008; tkaczuk et al. 2011b) in a range of habitats, mainly in phytocoenoses of lime-oakhornbeam forest (tilio cordatae-carpinetum betuli), continental swamp/bog pine forest (vaccinio uliginosi-pinetum), as well as in patches of sand grasslands (koelerio glaucae-corynephoretea class) covering inland dunes. fungi were also gathered from coniferous, deciduous and mixed forests (without determination of their syntaxonomic affinity) and from other habitats, including forest edges, roadsides, pathways, meadows, etc. the substrate of every species was determined and listed together with locality, date of collection and collector/identifier name(s). standard methods for studying each taxonomic group were used in the species identification (cf. dynowska, ejdys 2011; keller 1987; bałazy 1993; goettel, inglis, 1997; budziszewska et al. 2010). identification was based on microscopic analyses using light microscope with ‘phase contrast’ and magnification 100-1000x and handmade sections or squash preparations mounted in water, lactophenol blue, lactophenol picric acid solution or aceto-orcein. the soil fungi were isolated by the soil dilution plate method on potato dextrose agar, the bait method, using dry shrimps and gammarids as baits, to obtain species belonging to kickxellales (bills et al. 2004; kurihara et al. 2008) and warcup soil plate technique (warcup 1950). some saprobic species were obtained directly from organic material stored in moist chambers (kurihara, degawa 2006). the following monographs and keys were used: braun (1995a, b, 1998), domsch et al. (1993), ellis, ellis (1997), gams (1977), hoffmann et al. (2007), ignatavičiutė, treigienė (1998), kochman, majewski (1970, 1973), majewski (1977, 1979), nannfeldt (1981), sałata (1985), skirgiełło, zadara (1979), vanev et al. (1997), watanabe (2002), wołczańska (2005), zheng, chen (2001) and zycha et al. (1969). host plant species were determined using the key by rutkowski (2004). taxonomic system of fungi and fungus-like organisms of kirk et al. (2008) is adopted while nomenclature follows keller (2007), mułenko et al. (2008), and index fungorum. names of plants 216 m. ruszkiewicz-michalska et al. and arthropods are given after mirek et al. (2002) and schaefer (2010), respectively, while syntaxonomic nomenclature follows matuszkiewicz (2006). the specimens documenting research are deposited in the fungal collection of the herbarium universitatis lodziensis (lod), in the herbarium of the faculty of biology, university of warsaw (wa) as well as in fungal collection of department of plant protection of siedlce university of natural sciences and humanities, and in the department of mycology of the warmia and mazury university in olsztyn. results in total, 188 fungal taxa were identified as the result of four days of field surveying. a number of interesting specimens still await determination. the recorded species belong to true fungi: 33 zygomycetes (entomophthorales, kickxellales, mortierellales, mucorales, zoopagales), 130 ascomycetes (capnodiales, dothideales, erysiphales, hypocreales, phyllachorales, pleosporales, xylariales, anamorphs) and 22 basidiomycetes (exobasidiales, pucciniales, urocystales), as well as to chromistan (peronosporales) and protozoan (dictyostelida) fungal analogues (tab. 1). the predominant group (135 species) comprises micromycetes associated with aboveground plant organs and other fungi. fungi isolated from the soil (36 species) are the next numerous group and arthropod pathogens (17) are less numerous. the following abbreviations and symbols are used in the lists of species: kpu – kapice protective unit; gpu – grzędy protective unit; wpu – werykle protective unit; t-c – tilio cordatae-carpinetum betuli; v-p – vaccinio uliginosi-pinetum; (a) – anamorphic stage; (t) – teleomorphic stage; (0, i, ii, iii) – successive stages of rust fungi; * – species new to poland. initials of the names are given to denote collecting and/or identifying person (e.g., js – jarosław szkodzik). table 1 numbers of species of fungi and fungus-like organisms collected at the study area g ro up o f f un gi s .l. an am or ph ic fu ng i1 c ap no di al es d ic ty os te lid a d ot hi de al es e nt ho m op ht ho ra le s e ry si ph al es e xo ba si di al es h yp oc re al es ( in cl . a na m or ph s) k ic kx el la le s m or ti er el la le s m uc or al es pe ro no sp or al es p hy lla ch or al es p le os po ra le s p uc ci ni al es u ro cy st al es x yl ar ia le s z oo pa ga le s in to ta l no. 79 4 1 1 6 22 1 13 2 6 14 2 1 2 20 1 8 5 188 1 species with uncertain or unknown anamorph-teleomorph connection, presumably ascomycetes micromycetes in the biebrza national park 217 micromycetes associated with plants. among 135 taxa identified, the most numerous were anamorphs of ascomycetes (71 species), while pucciniales (20) and erysiphales (22) are represented by a lower number of species. other taxa (capnodiales, dothideales, exobasidiales, hypocreales, peronosporales, phyllachorales, pleosporales, urocystales, xylariales) contribute to a lesser extent to the list. two species are new in polish mycobiota: alternaria nobilis and metasphaeria cumana. the majority of taxa enumerated below are common in poland (cf. mułenko et al. 2008), with the exception of asteromella eupatoriicola, cercospora berteroae, ectostroma iridis, fusicladium scribnerianum, exobasidium rostrupii, mycosphaerella iridis, paraconiothyrium tiliae and septogloeum carthusianum, each known from a single locality. the other six species are rarely recorded in poland. the highest species richness of micromycetes associated with plants was observed in patches of tilio-carpinetum (65 species), while sand grassland (koelerio glaucae-corynephoretea class) and vaccinio uliginosi-pinetum hosted lower number of taxa (24 and 9 species, respectively). recorded fungi species predominantly occupied leaves and only a small number occurred on stems, flowers and fruits. logs, stumps and fallen branches of deciduous trees were colonized by xylariales and hypocreales members. among them the fungi of an early stage of wood decomposition (e.g., diatrypella favacea, d. quercina, hypoxylon fuscum, nectria cinnabarina), as well as the saprotrophs of the later stages (annulohypoxylon multiforme, nemania serpens, xylaria longipes), were distinguished. in addition, 7 taxa of fungicolous fungi were found: dialonectria episphaeria on old stromata of diatrypella favacea and alternaria sp. on the mycelium of an unidentified powdery mildew, as well as hyperparasites of erysiphales (ampelomyces quisqualis, phoma glomerata) and pucciniales (ramularia coleosporii, r. uredinearum, sphaerellopsis filum). acremonium sp. s.l. on equisetum pratense ehrh., kpu, t-c, 28 aug., leg. & det. mrm. *alternaria nobilis (vize) simmons [= a. dianthi stevens & hall] on dianthus carthusianorum l., gpu, dune, sand grassland, 30 aug., leg. mrm, det. mrm & es. alternaria sp. on mycelium of an unidentified, sterile powdery mildew (on quercus petraea (matt.) liebl. x q. robur l.), kpu, t-c, glade, 28 aug., leg. & det. mrm. ampelomyces quisqualis ces. s.l. on microsphaera divaricata (on frangula alnus), gpu, v-p, 29 aug., leg. & det. mrm; on golovinomyces sordidus (on plantago intermedia), kpu, t-c, edge of forest, 28 aug., leg. & det. mrm; on neoerysiphe galeopsidis (on galeobdolon luteum), kpu, t-c, 28 aug., leg. & det. mrm. annulohypoxylon multiforme (fr.) y.m. ju, j.d. rogers & h.m. hsieh on log of deciduous tree, gpu, mixed forest, 29 aug., leg. & det. js. ascochyta stellariae fautrey on stellaria holostea l., kpu, t-c, glade, 28 aug., leg. & det. mrm. ascochyta tenerrima sacc. & roum. on viburnum opulus l., gpu, mixed forest, 29 aug., leg. mrm, det. es & mrm. asteroma padi dc. on padus avium mill., kpu, t-c, 28 aug., leg. & det. mrm. note. rare species, known from two records from southern and western part of poland (mułenko et al. 2008). 218 m. ruszkiewicz-michalska et al. asteromella corcontica (kabát & bubák) moesz ex bat. & peres on hieracium sp. sect. eu-hieracium, gpu, dune, sand grassland, 30 aug., leg. md, det. mrm & es. asteromella eupatoriicola (kabát & bubák) ruppr. on eupatorium cannabinum l., gpu, v-p, roadside, 29 aug., leg. & det. mrm. note. species know from a single locality in central poland (ruszkiewicz-michalska 2006; mułenko et al. 2008). asteromella quercifolii c. massal. on quercus robur l., gpu, deciduous forest, 29 aug., leg. & det. es. asteromella tiliicola (oudem.) arx on tilia cordata mill., kpu, t-c, 28 aug., leg. & det. mrm. asteromella trautmanniana (moesz) moesz on sorbus aucuparia l. emend. hedl , gpu, mixed forest, 29 aug., leg. & det. mrm; deciduous forest, 30 aug., leg. & det. mrm. cercospora berteroae hollós on berteroa incana (l.) dc., gpu, dune, sand grassland, 30 aug., leg. & det. md & es; same host, locality and date, leg. & det. mrm. notes. species only recently reported as new to polish biota (połeć, ruszkiewicz-michalska 2012), collected primarily on the same host in the łódź city in 2006. according to the newest molecular studies (groenewald et al. 2012), c. berteroae is a synonym of cercospora armoraciae sacc., a species common in poland (mułenko et al. 2008). cercospora mercurialis pass. on mercurialis perennis l., kpu, t-c, 28 aug., leg. & det. mrm. cladosporium sp. s.l. on acinos arvensis (lam.) dandy, kpu, glade with tall herbs, 28 aug., leg & det. mrm; on anthoxanthum odoratum l., kpu, t-c, 28 aug., leg. & det. md; on equisetum pratense ehrh., kpu, t-c, 28 aug., leg. & det. mrm; on fragaria vesca l., kpu, glade with tall herbs, 28 aug., leg. & det. mrm; on quercus robur l., gpu, deciduous forest, 29 aug., leg. & det. es. coleosporium tussilaginis (pers.) lév. (iii, iii) on campanula sp., gpu, mixed forest, 29 aug., leg. & det. mrm; on melampyrum nemorosum l., kpu, t-c, 28 aug., leg. & det. mrm; gpu, deciduous forest, glade, 29 aug., leg. & det. mrm; gpu, dune, edge of sand grassland, 30 aug., leg. es, det. md & es. colletotrichum dematium (pers.) grove on asarum europaeum l., kpu, t-c, 28 aug., leg. md, det. md & es; on maianthemum bifolium (l.) f.w. schmidt, kpu, t-c, 28 aug., leg. & det. mrm; on polygonatum odoratum (mill.) druce, gpu, mixed forest, 29 aug., leg. & det. mrm. colletotrichum gloeosporioides (penz.) penz. & sacc. on anthoxanthum odoratum l., kpu, t-c, 28 aug., leg. md, det. md & es. note. a. odoratum is a new host for the fungus in poland. coniothyrium olivaceum bonord. [= microsphaeropsis olivacea (bonord.) höhn.] on sorbus aucuparia l. emend. hedl., gpu, deciduous forest, 30 aug., leg. & det. mrm. cronartium flaccidum (alb. & schwein.) winter (iii) on vincetoxicum hirundinaria medik., gpu, mixed forest, 29 aug., leg. & det. mrm. dialonectria episphaeria (tode) cooke on old stromata of diatrypella favacea, kpu, t-c, 28 aug., leg. & det. js. micromycetes in the biebrza national park 219 diatrype stigma (hoffm.) fr. on fallen twigs of corylus avellana l., kpu, t-c, 28 aug., leg. & det. js. diatrypella favacea (fr.) ces. & de not. on fallen twigs of corylus avellana l., kpu, t-c, 28 aug., leg. & det. js; on fallen twigs of deciduous tree, gpu, 29 aug., leg. & det. js. diatrypella quercina (pers.) cooke on fallen branches of quercus robur l., kpu, t-c, 28 aug., leg. & det. js. discosia artocreas (tode) fr. on betula pubescens ehrh., gpu, mixed forest, 29 aug., leg. & det. mrm. discosia minuta ces. on polygonum mite schrank, kpu, t-c, 28 aug., leg. & det. mrm; on vaccinium myrtillus l., gpu, v-p, 29 aug., leg. & det. mrm. note. species rare in poland (wołczańska et al. 2004). discosia sp. on fallopia convolvulus (l.) a. löve, kpu, t-c, 28 aug., leg. md, det. md & es. discula betulina (westend.) arx on betula pendula roth, kpu, t-c, 28 aug., leg. & det. mrm. discula umbrinella (berk. & broome) morelet on quercus petraea (matt.) liebl. x q. robur l., kpu, t-c, glade, 28 aug., leg. & det. mrm. ectostroma iridis (ehrenb.) fr. on iris pseudoacorus l , kpu, t-c, 28 aug., leg. & det. mrm. note. species know from a single locality in north eastern poland (durska 1974; mułenko et al. 2008). epicoccum nigrum link on stellaria holostea l., kpu, t-c, 28 aug., leg. & det. md; on quercus robur l., gpu, deciduous forest, 29 aug., leg & det. es. erysiphe adunca (wallr.) fr. [= uncinula adunca (wallr.) lév.], (t), on salix caprea l., gpu, mixed forest, 29 aug., leg. es, det. md & es; on salix sp., gpu, edge of reeds’ patch, near catwalk, 29 aug., leg. & det. mrm; path near dune, 30 aug., leg. mrm, det. es; deciduous forest, 30 aug., leg. & det. mrm. erysiphe alphitoides (griffon & maubl.) u. braun & s. takam. [= microsphaera alphitoides griffon & maubl.], (t), on quercus robur l., gpu, v-p, near catwalk, 29 aug., leg. & det. mrm. erysiphe aquilegiae dc. var. ranunculi (grev.) zheng & chen, (t), on ranunculus acris l., gpu, mixed forest, pathway, 29 aug., leg. & det. mrm. erysiphe convolvuli dc., (t), on convolvulus arvensis l., gpu, dune, sand grassland, 30 aug., leg. & det. md. erysiphe euonymi dc. [= microsphaera euonymi (dc.) sacc.], (t), on euonymus europaea l., kpu, t-c, 28 aug., leg. & det. mrm; gpu, mixed forest, 29 aug., leg. & det. mrm. erysiphe heraclei dc., (a), on peucedanum oreoselinum (l.) moench., gpu, dune, sand grassland, 30 aug., leg. & det. es; on pimpinella major (l.) huds. (?), gpu, deciduous forest, 30 aug., leg. mrm, det. es. erysiphe howeana u. braun, (a), on oenothera biennis l. s.l., gpu, dune, sand grassland, 30 aug., leg. & det. mrm. 220 m. ruszkiewicz-michalska et al. erysiphe hypophylla (nevod.) u. braun & cunningt. [= microsphaera hypophylla nevod.], (t), on quercus robur l., gpu, deciduous forest, 29 aug., leg. & det. es. erysiphe ornata var. europaea (u. braun) u. braun & s. takam. [= microsphaera ornata var. europaea u. braun], (t), on betula pendula roth, kpu, t-c, 28 aug., leg. & det. mrm; on betula pubescens ehrh., gpu, v-p, near catwalk, 29 aug., leg. & det. mrm; same locality, mixed forest, 29 aug., leg. & det. mrm. erysiphe tortilis (wallr.) link [= microsphaera tortilis (wallr.) speer], (a), on cornus sanguinea l., gpu, mixed forest, roadside, 29 aug., leg. & det. md. erysiphe ulmariae pers. ex desm., (t), on filipendula ulmaria (l.) maxim., gpu, willow thickets, near catwalk, 29 aug., leg. & det. mrm. exobasidium rostrupii nannf. on oxycoccus palustris pers., gpu, v-p, near catwalk, 29 aug., leg. & det. mrm; same locality and date, leg. grażyna domian, det. mrm. note. species known in poland from a single locality in poleski national park (mułenko 1988; mułenko et al. 2008). fusarium sp. on daphne mezereum l., gpu, mixed forest, 29 aug., leg & det. mrm; on knautia arvensis (l.) coult, kpu, t-c, 28 aug., leg. & det. md; on stellaria holostea l., kpu, t-c, 28 aug., leg. & det. md. fusicladium scribnerianum (briosi & cavara) m.b. ellis on betula pendula roth, kpu, t-c, 28 aug., leg. & det. mrm. note. species known only on betula pendula roth in a single localty in central poland (ruszkiewiczmichalska, połeć 2006; mułenko et al. 2008). golovinomyces sordidus (junell) heluta [= erysiphe sordida junell], (a), on plantago intermedia gillib., kpu, t-c, edge of forest, 28 aug., leg. & det. mrm. gymnosporangium cornutum arth. ex kern (0, i) on sorbus aucuparia l. emend. hedl., gpu, mixed forest, 29 aug. leg. & det. mrm; deciduous forest, 30 aug., leg. & det. mrm. hypoxylon fuscum (pers.) fr. on wood of corylus avellana l., gpu, mixed forest, 29 aug., leg. & det. js. leptosphaeria sp. on stellaria holostea l., kpu, t-c, glade, 28 aug., leg. & det. mrm. note. the morphology of the fungus is similar to the one described by mułenko and kozłowska (2010) who repeteadly collected it in the białowieża national park. leveillula taurica (lév.) arnaud, (t), on helichrysum arenarium (l.) moench, gpu, dune, sand grassland, 30 aug., leg. & det. md. melampsora epitea thüm. (ii, iii) on salix cinerea l , kpu, meadow close to goniądz village, willow thickets, 28 aug., leg. & det. mrm; on salix sp., gpu, path near dune, 30 aug., leg. mrm, det. es; on salix viminalis l., gpu, willow thickets at a roadside, 30 aug., leg. md, det. md & es. melampsora populnea (pers.) karst. (ii, iii) on populus tremula l., gpu, mixed forest, 29 aug., leg. & det. mrm. melampsoridium betulinum (pers.) kleb. (ii, iii) on betua pendula roth, kpu, t-c, 28 aug., leg. & det. mrm; on betula pubescens ehrh., kpu, t-c, 28 aug., leg. & det. mrm; gpu, v-p, near catwalk, 29 aug., leg. & det. mrm; same locality, mixed forest, 29 aug., leg. & det. mrm. micromycetes in the biebrza national park 221 melasmia acerina lév. on acer negundo l., osowiec-twierdza, roadside, 31 aug., leg. md, det. es; on acer pseudoplatanus l., kpu, t-c, 28 aug., leg. & det. mrm; gpu, mixed forest, 29 aug., leg. & det. mrm. melasmia punctata sacc. & roum. on acer platanoides l., kpu, t-c, 28 aug., leg. & det. mrm. melasmia sp. on salix caprea l., gpu, willow thickets near pathway, 30 aug., leg. & det. es. *metasphaeria cumana (sacc. & speg.) sacc. f. macrospora fautrey on carex hirta l., kpu, mid-forest damp meadow, 28 aug., leg. & det. mrm. microsphaera divaricata (wallr.) lév. on frangula alnus mill., (a), kpu, t-c, 28 aug., leg. & det. mrm; (t), gpu, v-p, 29 aug., leg. & det. mrm. mycosphaerella epilobii-montani lobik on epilobium palustre l., gpu, mixed forest, 29 aug., leg. & det. mrm. mycosphaerella iridis (auersw.) j. schröt. on iris pseudoacorus l., kpu, t-c, 28 aug., leg. & det. mrm. note. species known in a single localty in northeastern poland (adamska 2001; mułenko et al. 2008). mycosphaerella isariphora (desm.) johanson on stellaria holostea l., kpu, t-c, glade, 28 aug., leg. & det. mrm. mycosphaerella punctiformis (pers.) starbäck on quercus petraea (matt.) liebl. x q. robur l., kpu, t-c, glade, 28 aug., leg. & det. mrm; in anamorphic stage (as phyllosticta betulina sacc.) on betula pubescens ehrh., gpu, mixed forest, 29 aug., leg. & det. mrm. nectria cinnabarina (tode) fr. on fallen twigs, kpu, t-c, 28 aug., leg. & det. mw; on wood of deciduous trees, gpu, mixed forest, 29 aug., leg. & det. mw. nemania serpens (pers.) gray on fallen branches of deciduous trees, gpu, 29 aug., leg. & det. js. neoerysiphe galeopsidis (dc.) u. braun [= erysiphe galeopsidis dc.], (a), on galeobdolon luteum huds., kpu, t-c, 28 aug., leg. & det. mrm; gpu, mixed forest, 29 aug., leg. & det. mrm; on galeopsis tetrahit l., kpu, t-c, 28 aug., leg. & det. md; gpu, deciduous forest, pathway, 30 aug., leg. & det. mrm; edge of dune, 30 aug., leg. & det. mrm. ochropsora ariae (fuckel) ramsb. (ii, iii) on sorbus aucuparia l. emend. hedl., gpu, mixed forest, 29 aug., leg. & det. mrm. paraconiothyrium tiliae (f. rudolphi) verkley & gruyter [= asteroma tiliae f. rudolphi] on tilia cordata mill., kpu, t-c, 28 aug., leg. & det. mrm. passalora campi-silii (speg.) u. braun on impatiens noli-tangere l., gpu, mixed forest, 29 aug., leg & det. mrm. passalora ferruginea (fuck.) u. braun & crous on artemisia vulgaris l., kpu, t-c, forest edge, 28 aug., leg. & det. md; same locality and date, leg. & det. mrm. passalora microsora (sacc.) u. braun on tilia cordata mill., gpu, mixed forest, 29 aug., leg. & det. mrm; gpu, deciduous forest edge, 30 aug., leg. & det. mrm. 222 m. ruszkiewicz-michalska et al. peronospora alsinearum casp., (a), on stellaria media (l.) vill., gpu, deciduous forest, glade, 30 aug., leg. & det. mrm. peronospora sordida berk., (a, t), on scrophularia nodosa l., kpu, t-c, 28 aug., leg. & det. mrm. pestalosphaeria sp. on dianthus cartusianorum l , leg. mrm, det. mrm & es. note. so far, no member of the genus has been reported in poland (mułenko et al. 2008). phaeosphaeria sp. on sorbus aucuparia l. emend. hedl., gpu, deciduous forest, 30 aug., leg. mrm, det. mrm & es. phoma glomerata (corda) wollenw. & hochapfel on neoerysiphe galeopsidis (on galeopsis tetrahit), gpu, edge of dune, 30 aug., leg. & det. mrm; on phyllactinia guttata (on betula pendula), gpu, edge of dune, 30 aug., leg. & det. mrm. phragmidium potentillae (pers.) p. karst. on potentilla sp., gpu, dune, sand grassland, 30 aug., leg. & det. es. phyllachora graminis (pers.) fuck. on poa sp., kpu, t-c, 28 aug., leg. & det. md. phyllactinia guttata (wallr.) lév., (t), on betula pendula roth, gpu, edge of dune, 30 aug., leg. & det. mrm. phyllosticta aceris saccardo on acer negundo l., osowiec-twierdza, roadside, 31 aug., leg. md, det. es. notes. a. negundo is a new host for the fungus in poland (mułenko et al. 2008). phyllosticta campestris pass. on acer negundo l., osowiec-twierdza, roadside, 31 aug., leg. md, det. es. note. a. negundo is a new host for the fungus in poland (mułenko et al. 2008). phyllosticta cruenta (kunze: fries) kickx on convallaria majalis l., gpu, t-c, 29 aug., leg. & det. md. phyllosticta erysimi west. on alliaria petiolata (bieb.) cavara & grande, kpu, t-c, 28 aug., leg. & det. md. note. species known from a single locality in central poland (ruszkiewicz-michalska 2006; mułenko et al. 2008). phyllosticta euonymella sacc. on euonymus europaea l., kpu, t-c, 28 aug., leg. & det. mrm. phyllosticta lysimachiae allesch. on lysimachia vulgaris l., kpu, t-c, 28 aug., leg. & det. mrm. phyllosticta passerinii berl. & voglino on padus avium mill., kpu, t-c, 28 aug., leg. & det. mrm. phyllosticta polygonorum sacc. on fallopia convolvulus (l.) á. löve, gpu, dune, sand grassland, 30 aug., leg. md, det. mrm & es; on polygonum mite schrank, kpu, t-c, 28 aug., leg. & det. mrm. podosphaera balsaminae (wallr.) u. braun & s. takam. [= sphaerotheca balsaminae (wallr.) kari] on impatiens noli-tangere l., (a), kpu, t-c, 28 aug., leg. es, det. md & es; (t), same locality and date, leg. & det. mrm; on impatiens parviflora dc., kpu, t-c, 28 aug., leg. & det. md. micromycetes in the biebrza national park 223 podosphaera clandestina (wallr.) lév. var. aucupariae (erikss.) u. braun, (t), on sorbus aucuparia l. emend. hedl., gpu, mixed forest, 29 aug., leg. & det. mrm. podosphaera fuliginea (schltdl.) u. braun & s. takam. [= sphaerotheca fuliginea (schltdl.) pollacci], (t), on veronica chamaedrys l., kpu, t-c, 28 aug., leg. & det. md. podosphaera myrtillina (schubert) kunze var. major juel, (t), on vaccinium uliginosum l., gpu, v-p, near catwalk, 29 aug., leg. & det. mrm. podosphaera tridactyla (wallr.) de bary, (t), on padus avium mill., kpu, t-c, 28 aug., leg. & det. mrm. podosphaera xanthii (castagne) braun & schiskoff, (t), on melampyrum nemorosum l., kpu, t-c, 28 aug., leg. mrm, det. mrm & es. puccinia arenariae (schum.) wint., (iii), on melandrium album (mill.) garcke, kpu, t-c, forest edge, 28 aug., leg. & det. mrm; on stellaria media (l.) vill., gpu, deciduous forest, glade, 30 aug., leg. & det. mrm; kpu, t-c, 28 aug., leg. & det. mrm. puccinia asarina kunze, (iii), on asarum europaeum l., kpu, t-c, 28 aug., leg. md, det. md & es; same locality and date, leg. & det. mrm; gpu, mixed forest, 29 aug., leg. & det. mrm. puccinia chaerophylli purton, (ii, iii), on chaerophyllum bulbosum l. (?), kpu, damp glade with tall herbs, 28 aug., leg. & det. mrm. puccinia cnici-oleracei pers. ex desm., (iii), on artemisia vulgaris l., gpu, dune, sand grassland, 30 aug., leg. es, det. md & es. puccinia impatientis schubad, (ii, iii), on impatiens noli-tangere l., kpu, t-c, 28 aug., leg. & det. md; same locality and date, leg. & det. mrm. puccinia lapsanae fuck., (ii, iii), on lapsana communis l., kpu, t-c, 28 aug., leg. & det. mrm. puccinia polygonii alb. et schw., (ii, iii), on fallopia convolvulus (l.) á. löve, gpu, deciduous forest, 29 aug., leg. & det. es; same locality, dune, sand grassland, 30 aug., leg. & det. mrm; on fallopia dumetorum (l.) holub, kpu, t-c, 28 aug., leg. & det. mrm. puccinia punctata link, (ii), on galium sylvaticum l., gpu, mixed forest, 29 aug., leg. & det. es; on galium verum l., kpu, glade with tall herbs, 28 aug., leg. & det. mrm. puccinia tanaceti dc., (ii, iii), on artemisia vulgaris l., osowiec-twierdza, roadside, 31 aug., leg. & det. es. puccinia violae dc., (iii), on viola sp., gpu, mixed forest, 29 aug., leg. & det. mrm. pucciniastrum agrimoniae (dietel) tranzsch., (ii), on agrimonia eupatoria l., kpu, t-c, 28 aug., leg. & det. mrm. ramularia agrimoniae sacc. on agrimonia eupatoria l., kpu, t-c, glade, 28 aug., leg. & det. mrm. note. rare species, known from two localities in eastern and southeastern part of poland (wołczańska 2005). 224 m. ruszkiewicz-michalska et al. ramularia chamaedryos (lindr.) gunnerb. on veronica chamaedrys l., kpu, glade with tall herbs, 28 aug., leg. & det. mrm. ramularia coleospori sacc. on coleosporium tussilaginis (on melampyrum nemorosum), kpu, t-c, 28 aug., leg. & det. mrm. ramularia grevilleana (tul.) jørst. [= r. arvensis sacc.] on potentilla sp., gpu, dune, sand grassland, 30 aug., leg. & det. es. ramularia inaequale (preuss) u. braun on hieracium sp. sect. eu-hieracium, gpu, dune, sand grassland, 30 aug., leg. md, det. mrm & es. ramularia lapsanae (desm.) sacc. on lapsana communis l., gpu, mixed forest, 29 aug., leg. & det. es; same locality and date, leg. & det. mrm. ramularia pratensis sacc. on rumex sp., kpu, glade with tall herbs, 28 aug., leg & det. mrm. ramularia tricheriae lindroth on knautia arvensis (l.) coult, kpu, t-c, glade, 28 aug., leg. & det. mrm; gpu, dune, sand grassland, 30 aug., leg. md, det. mrm & es. ramularia uredinearum hulea on telia of puccinia arenariae (on stellaria holostea), gpu, mixed forest, 30 aug., leg. & det. mrm. note. this is the first confirmed collection of the species as the specimens documenting its previous reports do not exsist (wołczańska 2005). ramularia urticae ces. on urtica dioica l., kpu, t-c, 28 aug., leg & det. mrm; gpu, willow thickets, near catwalk, 29 aug., leg. & det. mrm; same locality and date, leg. & det. es. ramularia variabilis fuckel on verbascum nigrum l., kpu, glade with tall herbs, 28 aug., leg. & det. mrm. seimatosporium lichenicola (corda) shoemaker & müll. on pyrus communis l , gpu, dune, sand grassland edge, near the pathway, 30 aug., leg. mrm, det. mrm & es. septogloeum carthusianum (sacc.) sacc. on euonymus europaea l., kpu, t-c, 28 aug., leg. & det. mrm. note. species very rare in poland, known from signle report from the noteć river valley in ne poland (michalski 1982; mułenko et al. 2008). septoria aegopodii desm. ex kickx on aegopodium podagraria l., gpu, mixed forest, 29 aug., leg. & det. es; same locality and date, leg. & det. mrm. septoria betulae pass. on betula pendula roth, gpu, edge of dune, 30 aug., leg. & det. mrm. septoria chelidoni (lib.) desm. on chelidonium majus l., kpu, t-c, 28 aug., leg. & det. mrm; gpu, dune, edge of sand grassland, 30 aug., leg. & det. md & es. septoria dulcamarae desm. on solanum dulcamara l., gpu, willow thickets, near catwalk,, 29 aug., leg. & det. mrm. septoria hyperici desm. on hypericum perforatum l ,kpu, glade with tall herbs, 28 aug., leg. mrm, det. es. septoria lychnidis desm. on melandrium album (mill.) garcke, gpu, dune, edge of sand grassland, 30 aug., leg. & det. mrm. micromycetes in the biebrza national park 225 septoria oenotherae west. on oenothera biennis l. s.l., gpu, dune, sand grassland, 30 aug., leg. & det. es; same locality and date, leg. & det. mrm. septoria polygonorum desm. on fallopia convolvus (l.) á. löve, gpu, dune, sand grassland, 30 aug., leg. md, det. mrm & es. septoria pyricola desm. on pyrus communis l., gpu, dune, sand grassland edge, near the pathway, 30 aug., leg. mrm, det. es & mrm. septoria sedi westend. on sedum telephium l. (?), gpu, dune, sand grassland, 30 aug., leg. mrm, det. mrm & es. septoria virgaureae (lib.) desm. on solidago virgaurea l., gpu, dune, sand grassland, 30 aug., leg. & det. md & es. sphaerellopsis filum (biv.) b. sutton on melampsora epitea (on salix cinerea), kpu, meadow close to goniądz village, willow thickets, 28 aug., leg. & det. mrm; (on salix viminalis) gpu, willow thickets at a roadside, 30 aug., leg. md, det. es; on puccinia chaerophylli (on chaerophyllum bulbosum), kpu, damp glade with tall herbs, 28 aug., leg. & det. mrm; on puccinia tanaceti (on artemisia vulgaris) osowiec-twierdza, roadside, 31 aug., leg. & det. es. sphaeropsis sp. on dianthus carthusianorum l., gpu, dune, sand grassland, 30 aug., leg. & det. mrm. triphragmium ulmariae (dc.) link (ii, iii) on filipendula ulmaria (l.) maxim., kpu, t-c, edge of damp meadow, 28 aug., leg. & det. mrm. urocystis syncocca (kirchner) lind. on hepatica nobilis schreb., gpu, mixed forest, 29 aug., leg. & det. mrm. valdensia heterodoxa peyronel on betula pubescens ehrh., gpu, mixed forest, 29 aug., leg. & det. mrm. volutella ciliata (alb. & schwein.) fr. on impatiens parviflora dc., kpu, t-c, roadside, 28 aug., leg. & det. es. note. i. parviflora is a new host for the fungus in poland (mułenko et al. 2008). xylaria longipes nitschke on wood of deciduous trees, gpu, mixed forest, 29 aug., leg. js & dominika ślusarczyk, det. js. xylaria polymorpha (pers.) grev. on wood, osowiec-twierdza, area of the fortress, 31 aug., leg. & det. mw. micromycetes associated with arthropods. in total 15 species of entomopathogenic fungi were found as pathogens of different arthropods and three species were isolated from forest soil or litter by means of the galleria bait method. six entomophthoralean species from insects and one species from a mite were identified. the most numerous were species from the genus zoophthora. especially interesting is the second record of mycoses caused by zoophthora sp. 1. in a population of whiteflies (aleyrodes sp. l.) feeding on chelidonium majus l. for the first time we found that the pathogen can infect not only the adults (tkaczuk et al. 2011a) but also the larval stage of whiteflies. another fungal species from the genus zoophthora caused heavy epizootics in flies from the family lauxanidae in kapice and grzędy protected areas. the morphology of this fungus seems to be very close to z. radicans, but in the light of recent recommendations to verify species identity by molecular markers, 226 m. ruszkiewicz-michalska et al. this assumption should be verified. in the course of the study the spider mite eotetranychus tiliae has been identified as a new host for the acaropathogenic fungus neozygites floridana. from among 11 species of anamorphic hypocreales (ascomycota) affecting arthropods in investigated parts of the biebrza national park, two have been recognized as pathogens of spiders (gibellula leiopus and g. pulchra) and two were pathogenic to mites (hirsutella danubiensis and h. kirchneri). h. danubiensis is very rare and known only from single localities in poland and austria (bałazy et al. 2008; tkaczuk et al. 2011a). isaria farinosa and i. tenuipes, representing cordycipitaceous anamorphs, were found as pathogens infecting pupae of unidentified lepidoptera in the forest litter and soil. in addition, from both aforementioned environments the entomopathogenic fungi isaria fumosorosea, metarhizium anisopliae and beauveria bassiana were isolated by means of the galleria bait method. batkoa apiculata (thaxt.) humber on unidentified plant-hopper (homoptera), on the underside of corylus avellana l. leaf, gpu, deciduous forest, 29 aug., leg. & det. ct. beauveria bassiana (bals.-criv.) vuill. on unidentified bug (heteroptera), gpu, in the litter of deciduous forest, 29 aug., leg. & det. ct. furia sciarae (l.s. olive) humber on sciaridae flies, on the underside of leaves of different deciduous trees, gpu, 29 aug., leg. & det. ct. gibellula leiopus (vuill. ex maubl.) mains on spider from the family linyphiidae, on corylus avellana l. leaf, kpu, deciduous forest, 28 aug., leg. & det. ct. gibellula pulchra cavara on spider from the family thomisidae, gpu, deciduous forest, 29 aug., leg. js, det. ct. hirsutella danubiensis tkaczuk, bałazy & wegenst. on spider mite neotetranychus rubi (träg) feeding on rubus sp., kpu, deciduous forest, 28 aug., leg. & det. ct. hirsutella kirchneri (rostrup) minter, brady & hall on eriophyid mites (abacarus sp.) feeding on grass leaves, kpu, mid-forest clearing, 28 aug., leg. & det. ct. isaria farinosa (holm) brown & smith on unidentified pupa of lepidoptera, kpu, litter of deciduous forest, 28 aug., leg. mw, det. ct. isaria fumosorosea wize, isolated from the forest litter by means of galleria bait method, gpu, 29 aug., leg. & det. ct. isaria tenuipes peck on unidentified pupa of lepidoptera, kpu, litter of deciduous forest, 28 aug., leg. kamil kędra, det. ct. lecanicillium longisporum (petch) zare & w. gams on unidentified plant-hoppers and whiteflies feeding on different herbaceous plants, kpu, deciduous forest, 28 aug., leg. & det. ct. lecanicillium muscarium (petch) zare & w. gams on unidentified aphid, on the underside of acer l. leaf, kpu, deciduous forest, 28 aug., leg. & det. ct. metarhizium anisopliae (metschn.) sorokin s.l., isolated from the forest soil by means of galleria bait method, kpu, 28 aug., leg. & det. ct. micromycetes in the biebrza national park 227 neozygites floridana (j. weiser & muma) remaud. & keller on spider mite eotetranychus tiliae (hermann) feeding on tilia cordata mill. leaves, kpu, 28 aug., leg. & det. ct. note. eotetranychus tiliae is a new host for n. floridana. zoophthora aphrophorae (rostr.) s. keller on plant-hopper from the genus aphrophora feeding on potentilla argentea l., kpu, 28 aug., leg. & det. ct. zoophthora sp. 1. on adults and larvae of whiteflies of the genus aleyrodes l. feeding on chelidonium majus l., kpu, deciduous forest, 28 aug., leg. & det. ct. note. species known in poland from one locality (tkaczuk et al. 2011a). zoophthora sp. 2. on lauxaniidae flies on the underside of leaves of different deciduous trees, kpu, 28 aug., leg. & det. ct; gpu, 29 aug., leg. & det. ct. fungi isolated from soil and organic matter. in total 36 species of saprotrophic fungi were identified as a result of 33 plate analyses. 15 of them were prepared with the warcup method, 10 with gammari and shrimps baits, six plates with the dilution technique and two plates were filled with the excrement of wild animals. only one species, mucor hiemalis, was revealed in samples from several localities. kickxella alabastrina, a very rare species in poland, was observed only once, caught on a gammarus bait. it is worth noting that usually kickxellales members develop on a single bait unit, despite the regular arrangement of baits on soil entirely filling the plate. they are absent when the plate dilution method is used. the warcup method allows their disclosure, but they grow always from separate soil particles, which could be identified as insect excrement or insect remnants (kurihara 2006). absidia caerulea bainier, obtained with warcup method, kpu, deciduous forest, 28 aug., leg. & det. mw; wpu, „carska droga”, at road edge, spruce forest, 01 sept., leg. & det. mw. absidia glauca hagem, obtained with warcup method, wpu, „carska droga”, at road edge, birch forest, 01 sept., leg. & det. mw. absidia repens tiegh. s.l., obtained with warcup method, gpu, dune, sand grassland, 30 aug., leg. & det. mw. aspergillus candidus link on roe deer excrement, kpu, t-c, 28 aug., leg. & det. mw. cladosporium cladosporioides (fresen.) de vries, obtained with warcup method, wpu, barwik, peatbog, 31 aug., leg. & det. mw. *clonostachys solani f. solani (harting) schroers & w. gams on roe deer excrement, gpu, mixed forest, 29 aug., leg. & det. mw. coemansia sp., obtained with warcup method, kpu, t-c, 28 aug., leg. & det. mw. note. the species could not be identified with use of skirgiełło and zadara’s monograph (1979), nor with papers by kurihara et al. (2000, 2001, 2004, 2006, 2008). cunninghamella elegans lendn., obtained with warcup method, gpu, dune, sand grassland, 30 aug., leg. & det. mw. dictyostelium mucoroides bref., obtained with warcup method, wpu, „carska droga”, at road edge, spruce forest, 01 sept., leg. & det. mw. kickxella alabastrina coem., obtained with bait method, isolated on gammarus, kpu, deciduous forest, 28 aug., leg. & det. mw. 228 m. ruszkiewicz-michalska et al. lentamyces parricida (renner & muskat ex hesselt. & j.j. ellis) kerst., hoffmann & k. voigt, obtained with warcup method, kpu, deciduous forest, 28 aug., leg. & det. mw. *mariannaea elegans (corda) samson on spruce twig, kpu, mixed forest, 28 aug., leg. mw, det. mateusz wilk. mortierella bainieri costantin, obtained with bait method, isolated on gammarus, wpu, „carska droga”, at road edge, spruce forest, 01 sept., leg. & det. mw. mortierella bisporalis (thaxter) björling, obtained with bait method and observed directly on soil and plant remnants, kpu, deciduous forest, 28 aug., leg. & det. mw. mortierella polycephala coem., obtained with bait method, observed directly on soil and plant remnants, wpu, mixed forest, 31 aug., leg. & det. mw. mortierella zychae linnem., obtained with bait method, isolated on gammarus and observed directly on soil and plant remnants, wpu, mixed forest, 31 aug., leg. & det. mw. mucor fragilis bainier, obtained twice with bait method, isolated on gammarus, wpu, „carska droga”, at road edge, under wooden tower and from meadow soil, 01 sept., leg. & det. mw. mucor hiemalis wehmer, obtained with bait method and plate dilution method, gpu, alder fen forest, 30 aug., leg. & det. mw; dune, 30 aug., leg. & det. mw; wpu, „długa luka” educational path, meadow soil, 01 sept., leg. & det. mw. mucor mucedo l., obtained with warcup and bait methods, isolated on gammarus, wpu, mixed forest, isolated from the forest litter, 31 aug., leg. & det. mw. mucor plumbeus (bonord) arx, obtained with warcup method, wpu, „carska droga”, at road edge, birch forest, 28 aug., leg. & det. mw. mucor saturninus hagem, obtained with warcup method, gpu, dune, 30 aug., leg. & det. mw. penicillium commune thom, obtained with warcup method, wpu, „carska droga”, road edge, twice, from birch and spruce forests, 01 sept., leg. & det. mw. pilaira anomala (ces.) j. schröt., obtained with warcup method, kpu, t-c, 28 aug., leg. & det. mw. note. the fungus grew directly from soil particles, possibly from animal excrement. pilobolus crystallinus klein. on wild boar excrement, osowiec-twierdza, 31 aug., leg. & det. mw. piptocephalis lepidula (marchal) r.k. benj., obtained with warcup method and with plate dilution method, wpu, „długa luka” educational path, meadow soil, 01 sept., leg. & det. mw. piptocephalis sp., obtained with bait method, from soil particles, wpu, “carska droga”, at road edge, spruce forest, 01 sept., leg. & det. mw. note. this species with very thin branches and moist sporocladial heads has already been found several times in bbnp (boulahdjel 2010). rhizopus oryzae went & prins. geerl., obtained with warcup method, wpu, „długa luka” educational path, meadow soil, 01 sept., leg. & det. mw. micromycetes in the biebrza national park 229 rhizopus stolonifer (ehrenb.) vuill., obtained with warcup method, wpu, peatbog, 31 aug., leg. & det. mw. stilbella sp., directly on roe deer excrement, kpu, t-c, 28 aug., leg. & det. mw. syncephalis penicillata indoh., obtained with bait method, wpu, „długa luka” educational path, meadow soil, 01 sept., leg. & det. mw. syncephalis sphaerica tiegh., obtained with warcup method and with bait method, wpu, barwik, meadow soil, 31 aug., leg. & det. mw. syncephalis tenuis thaxter, obtained with bait method, wpu, barwik, meadow soil, 31 aug., leg. & det. mw. trichoderma koningii oudem., obtained with warcup method, kpu, t-c, 28 aug., leg. & det. mw; wpu, peatbog, 31 aug., leg. & det. mw. trichoderma viride pers., obtained with warcup method, and with plate dilution method, wpu, “carska droga”, at road edge, birch forest, 01 sept., leg. & det. mw. umbelopsis isabellina (oudem.) w. gams, obtained with warcup method, gpu, mixed forest, 29 aug., leg. & det. mw. umbelopsis ramanniana (möller) gams, obtained with warcup method, kpu, t-c, 28 aug., leg. & det. mw. final remarks the first, short-term and area-limited inventory of fungi occurring in the bbnp has yielded a relatively high number of fungi taxa. the ratio infected plants/parasitic fungi observed currently is also rather high, reaching 1:0.7 (cf. analysis by mułenko 1998, tab. 2). the presented results of our studies, although preliminary, give valuable information on the microfungi of the bbnp. the species’ richness and taxonomic diversity of the determined fungi, together with a number of yet unidentified specimens, is promising for future studies in the protected area of the biebrza valley. key outcomes: • the initiation of mycological studies in the bbnp, the area that until now had very fragmented information on the fungi present, comprising mainly species belonging to macromycetes and soil-inhabiting taxa. as a result of the study, basic ecological data on 188 micromycete taxa, classified in 17 orders and a group of anamorphic fungi, occurring in the area have been gathered. • reports of 128 taxa associated with plants, 17 species of arthropod pathogens, 12 species of soil fungi and 7 species of fungicolous fungi that are new to the mycobiota of the bbnp. • four species new in polish mycobiota (e.g., alternaria nobilis, clonostachys solani, mariannaea elegans, metasphaeria cumana) and 17 taxa very rarely recorded in poland, e.g., asteromella eupatoriicola, asteroma padi, cercospora berteroae, discosia minuta, ectostroma iridis, exobasidium rostrupii, fuscicladium scribne rianum, hirsutella danubiensis, kickxella alabastrina, mycosphaerella 230 m. ruszkiewicz-michalska et al. iridis, paraconiothyrium tiliae, phyllosticta aceris, p. erysimi, ramularia agrimoniae, r. uredinearum, septogloeum carthusianum, volutella ciliata, zoophtora sp. 1. acknowledgments. we are deeply grateful to mr. andrzej grygoruk (who at the time of our stay was the acting managing director) as well as to ms. magdalena maliszewska, ms. katarzyna nowicka, mr. cezary werpachowski and mr. krzysztof frąckiel, and other employees of the biebrza national park for collecting fungi, offering their facilities, helping in the implementation of our research and creating the very friendly atmosphere during our visit. we express our sincere gratitude to prof. maria rudawska (institute of dendrology pas in kórnik) for her valuable help and funding for linguistic corrections. we also thank ms. grażyna domian (regional directorate for environmental protection in szczecin), dr. dominika ślusarczyk (university of łódź) and mr. kamil kędra (polish mycological society) for kind permission to include their specimens into the list of species. sincere thanks are also addressed to anonymous reviewers for considerable remarks on the text. the studies were partially supported by the grant of the ministry of science and higher education no n n303 548839, by the university of warmia and mazury in olsztyn (grant no 02110802) and by the university of łódź (grant no 503/824). references adamska i. 2001. microscopic fungus-like organisms and fungi of the słowiński national park (nw poland). ii. acta mycol. 36 (1): 31–65. bałazy s. 1993. flora of poland. fungi (mycota). 24: entomophthorales. ed. polish academy of sciences, w. szafer institute of botany. kraków, 356 pp. bałazy s., miętkiewski r., tkaczuk c., wegensteiner r., wrzosek m. 2008. diversity of acaropathogenic fungi in poland and other european countries. exp. appl. acarol. 46: 53-70. doi:10.1007/s10493008-9207-1, http://link.springer.com/article/10.1007%2fs10493-008-9207-1. bills g.f., christensen m., powell m., thorn g. 2004. saprobic soil fungi. 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(in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. w. szafer institute of botany, polish academy of sciences, kraków: 53–70. wołczańska a. 2005. the ramularia species in poland. monogr. bot. 95: 1–154 http://www.ib-pan.krakow.pl/pubs-pdf/monographiae%20botanicae/2005/vol95.pdf [in polish with english summary]. wołczańska a., kozłowska m., piątek m., mułenko w. 2004. survey of the genus discosia (anamorphic fungi) in poland. polish bot. j. 49 (1): 55–61. http://www.ib-pan.krakow.pl/pubs-pdf/polish%20botanical%20journal/2004/pbj49-1_s055-61.pdf. zheng r. y., chen g. q. 2001. a monograph of cunninghamella. mycotaxon 80: 1–75. zycha h., siepmann r., linnemann g. 1969. mucorales eine beschreibung aller gattungen und arten dieser pilzgruppe. verlag von j. cramer, lehre. 234 m. ruszkiewicz-michalska et al. wstępne badania grzybów w biebrzańskim parku narodowym. i. grzyby mikroskopijne streszczenie prezentowane dane są wynikiem badań prowadzonych w dniach od 28 sierpnia do 1 września 2012 w ramach sesji terenowej polskiego towarzystwa mykologicznego. celem była wstępna inwentaryzacja grzybów i śluzowców występujących na terenie biebrzańskiego parku narodowego (bbpn). w artykule krótko podsumowano uzyskane wyniki, prezentowane w postaci adnotowanego wykazu 188 taksonów mikroskopijnej wielkości grzybów właściwych (33 gatunki grzybów sprzężniowych, 130 workowych i anamorficznych, 22 podstawkowe) oraz trzech gatunków zaliczanych do organizmów grzybopodobnych (peronospora alsinearum i p. sordida z chromista oraz dictyostelium mucoroides z protozoa). ogólem zidentyfikowano 128 gatunków związanych troficznie z roślinami, 17 ze stawonogami i 7 z grzybami oraz 36 gatunków grzybów wyizolowanych z gleby i szczątków organicznych. 89% wszystkich gatunków podano z terenu parku po raz pierwszy. wśród znalezionych grzybów są gatunki bardzo rzadko notowane (znane z 1-2 stanowisk) i cztery stwierdzone w polsce po raz pierwszy: alternaria nobilis, clonostachys solani, mariannaea elegans i metasphaeria cumana. pozostałe, nieuwzględnione tu gatunki mikromycetes nowe dla bioty polski będą scharakteryzowane w odrębnych artykułach. wyniki dotyczące makromycetes prezentowane są w części drugiej opracownia (kujawa et al. 2012). badania przeprowadzone w bbpn rozpoczynają planowany wieloletni cykl inwentaryzacji w obiektach chronionych, nieposiadających danych mykologicznych. celem tych badań jest uzyskanie danych będących podstawą do poznania biologii wielu gatunków, w tym ich reakcji na presję czlowieka. wiedza ta jest niezbędna zarówno do poznania roli grzybów mikroskopijnych w funkcjonowaniu obszarów chronionych i określenia zagrożeń jakim podlegają same grzyby, jak i do uświadomienia społeczeństwu konieczności ich ochrony na równi z innymi organizmami. 2014-01-02t12:16:32+0100 polish botanical society 2014-08-25t21:39:14+0200 polish botanical society 2014-08-28t17:49:09+0200 polish botanical society 2014-08-25t21:38:50+0200 polish botanical society 2014-08-29t18:49:51+0200 polish botanical society 2014-08-20t22:54:12+0200 polish botanical society © the author(s) 2014 published by polish botanical society diversity of macromycetes in the białaczów nature reserve (central poland) agnieszka salamaga1 and dominika ślusarczyk2 1jagiellonian university, institute of botany kopernika 27, pl-31-501 kraków, asalamaga@wp.pl 2university of lodz, faculty of biology and environmental protection department of algology and mycology, banacha 12/16, pl-90-237 łódź, dominika@biol.uni.lodz.pl salamaga a., ślusarczyk d.: diversity of macromycetes in the białaczów nature reserve (central poland). acta mycol. 49 (1): 99–107, 2014. the paper contains preliminary results of mycological research conducted in the białaczów forest reserve, situated at the wzgórza opoczyńskie upland. there are only data on macromycetes at this area. the occurrence of 167 species of macrofungi has been documented on the background of variety of tree stand and habitats. in july 2011 a heavy wind storm destroyed the tree stand and devastated the reserve. now the area is in course of natural renovation. the data presented in the paper could be used as comparative fungal material concerning qualitative and quantitative changes. key words: macrofungi, mycological research, rare species, protected area introduction the topic of the study are macrofungi in białaczów forest reserve, in reference to tree stands and habitat background. mycological studies in central poland were initiated by ławrynowicz (1973) and conducted in over 40 natural reserves (ławrynowicz 2002; szkodzik 2005; adamczyk 2007). the białaczów reserve is one of those reserves from which collections were not published jet. the fact that it was mostly destroyed during a wind storm in july 2011, led to the presentation of the existing material. the obtained preliminary results, and the gathered herbarium documentation will be the basis for comparison and confrontation of the changes that occurred in the fungal diversity after the storm. acta mycologica vol. 49 (1): 99–107 2014 doi: 10.5586/am.2014.012 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 100 a. salamaga and d. ślusarczyk © the author(s) 2014 published by polish botanical society study area the białaczów forest reserve (łódź province, opoczno forestry district, n 51°18, e 20°17) covers 21,87 ha and was created on 24.05.1976, it’s main goal being the preservation of a wet-ground forest with it’s natural characteristics (rąkowski 2006). the reserve contains a whole forestry unit (101). the borders of the reserve consist of a road leading from miedzna drewniana to białaczów in the south and west, a small watercourse with swampy meadows in the north, and lands belonging to the social house in białaczów in the east (fig. 1). in according to geographical classification, the reserve is located in wzgórza opoczyńskie upland, surrounding the świętokrzyskie mountains from north west. fig. 1. study area: 1 – the białaczów reserve, 2 – roads, 3 – forest sections, 4 – forest units. diversity of macromycetes in the białaczów nature reserve 101 © the author(s) 2014 published by polish botanical society the upland consist of jurassic rock, on which lay gravel monadnocks (kondracki 2002). in the reserve two types of soil are distinguished: muck-gleyed soil in the subunit a, and both brown and poor sand-based postglacial soil in the rest of the area (operat urządzeniowy 1998). the dominant plant community in the reserve is tilio-carpinetum calamagrostietosum and small fragments of natural riparian forests: fraxino-alnetum and ficarioulmetum. tree species occurring there are: alnus glutinosa, fraxinus excelsior, ulmus campestris, acer pseudoplatanus, a. platanoides, carpinus betulus. in tilio-carpinetum calamagrostietosum apart from species tilia cordata, fagus sylvatica, quercus robur and carpinus betulus, there are also those that were introduced into the treestand: betula pendula, pinus sylvestris, robinia pseudoacacia, quercus rubra and q. palustris. hepatica nobilis, a species under strict protection have been found in the reserve, and also species under partial protection: convallaria majalis, hedera helix, and pleurozium schreberi in the moss layer. material and methods mycological studies in the reserve were conducted in years 2006-2007, using the route method, and the permanent study plots. the following monographs and keys were used in fungi identification: skirgiełło (1960, 1991, 1998), domański (1965), jülich (1984), galli (1996), nespiak (1981, 1990), moser (1978), breitenbach, kränzlin (1984, 1986, 1991, 1995, 2000), knudsen, vesterholt (2008) and noordeloos et al. (2001, 2005). standard methods of studying macrofungi, based on microscopic analyses, were used in the species identification (hand-made sections or squash preparations mounted in water, 3-5% koh, melzer’s reagent; light microscope). the nomenclature follows wojewoda (2003), and index fungorum (2013). threat categories are listed according to wojewoda and ławrynowicz (2006). the specimens are deposited in the herbarium universitatis lodziensis (lod f). results and notes presented list contains first mycological data from wzgórza opoczyńskie upland. in total, 167 taxa of macomycetes (species and forms) including 13 ascomycota and 154 basidiomycota were identified. most of the gathered species were saprotrophic fungi – 118 species, which make 70% of the studied material. this shows an adequate amount of diverse substratum (leaves, needles, cones, small branches) needed for fungal growth in this habitat. 44 of gathered species were mycorrhizal fungi, and 5 were parasites. from ecological point of view, most of the gathered fungi produce fruit bodies above ground (74) and on wood (62), wheares fungi on plant litter were represented by 31 species. the following abbreviations are used in the list of species: t-c – tilio-carpinetum, f-u – ficario-ulmetum, f-a – ficario-almetum, r – red list category. 102 a. salamaga and d. ślusarczyk © the author(s) 2014 published by polish botanical society list of species ascomycota ascocoryne sarcoides (jacq.) j.w. groves & d.e. wilson – on wood, t-c, viii-xi 2006-2007. bisporella citrina (batsch) korf & s.e. carp. – on deciduous wood, t-c, v 2007. bulgaria inquinans (pers.) fr. – on deciduous wood, t-c, ix 2007. elaphomyces granulatus fr emend. holl. – in deciduous and mixed tree stands, in soil (ławrynowicz 1989). hypoxylon deustum (hoffm.) grev. – on dead wood, t-c, ix 2007. hypoxylon fragiforme (pers.) j. kickx – on dead wood, t-c, iii-xii 2006-20087 lasiosphaeria ovina (pers.) ces. & de not. – on dead wood, t-c, vii 2007. mollisia amenticola (sacc.) rehm – on wood, t-c, xi 2007. nectria cinnabarina (tode) fr. – on wood, t-c, f-u, f-a, viii 2007. otidea onotica (pers.) fuckel – on the ground, t-c, viii 2007. pachyphloeus melanoxanthus tul. – under fagus sylvatica, in soil (ławrynowicz 1990), r. scutellinia scutellata (l.) lambotte – on wood, t-c, vi 2006-2007. trichophaea hemisphaerioides (mouton) graddon – on the ground, t-c, ix 2007. xylaria hypoxylon (l.) grev. – on dead wood, t-c, ix-xi 2007,vi-xi 2007. xylaria polymorpha (pers.) grev. – on dead wood, t-c, x-xii 2006-2007. basidiomycota agaricus silvaticus schaeff. – on the ground, t-c, x 2007. agaricus silvicola (vittad.) peck. – on the ground, t-c, ix-x 2007. amanita citrina (schaeff.) pers. – on the ground, t-c, f-u, viii-x 2007. amanita citrina var. alba (gillet) gilbert. – on the ground, t-c, x 2007. amanita muscaria (l. fr.) hook. – on the ground, t-c, ix-x 2007. amanita pantherina (dc.) krombh. – on the ground, t-c, ix-x 2007. amanita porphyria (alb & schwein.: fr) mladý. – on the ground, t-c, ix 2007. armillaria mellea (vahl.: fr.) p. kumm. s.str. – on dead wood, t-c, x 2007. auricularia auricula-judae (bull.: fr.). wettst – on a branch of a deciduous tree, t-c, f-u, iii-xi 2007. auriscalpium vulgare gray – on pine cone, t-c, ix-xi 2007. boletus edulis bull.: fr. – on the ground, t-c, ix 2007. calocera viscosa (pers.: fr.) fr. – on conifers stump, t-c, vi-vii 2007. cantharellus cibarius fr. – on the ground, t-c, vii-x 2007. chroogomphus rutilus (schaeff.: fr.) o. k. miller – on the ground, t-c, x 2007. clavariadelphus fistulosus (holmsk.) corner – on twigs of deciduous tree, t-c, xi 2007, r. clavariadelphus junceus (alb & schwein.: fr.) corner – on fallen leaves of betula pendula, t-c, x 2007, r. clitocybe candicans (pers.) p. kumm. – on the ground, t-c, x 2007. clitocybe clavipes (pers.: fr.) p. kumm. – on the ground, t-c, x 2007. clitocybe gibba (pers.: fr.) p. kumm. – on the ground, t-c, x 2007. citocybe nebularis (batsch) p. kumm. – on the ground, f-u, x-xi 2007. clitocybe odora (bull.: fr.) p. kumm. – on the ground, t-c, viii 2007. clitocybe phyllophila ( fr.) p. kumm. – on litter, t-c, f-u, x 2007. coprinus micaceus (bull.: fr.) fr. – around stumps and logs of deciduous tree, t-c, f-u, x 2007. coprinus truncorum (scop.) fr. ss. romagn. – on stumps and logs on dead wood, f-u, vii 2007. cortinarius alboviolaceus (pers.: fr.) fr. – on the ground, t-c, x 2007. cortinarius hemitrichus (pers.: fr.) fr. – on the ground, t-c, x 2007. crepidotus variabilis (pers.: fr.) p. kumm. – on dead fallen twigs of deciduous tree, t-c, f-u, ix-x 2007. crucibulum laeve (huds.) kambly. – on a branch in the litter, t-c, ix 2007. cyathus striatus (huds.)willd. – on a branch in the litter, t-c, f-u, x 2007. cystoderma amianthinum (scop.: fr.) fayod – of mosses, t-c, x 2007. diversity of macromycetes in the białaczów nature reserve 103 © the author(s) 2014 published by polish botanical society dacrymyces stillatus nees: fr. – on the branch, t-c, f-u, v-xii 2007. daedaleopsis confragosa (bolt.: fr.) j. schröt. – on a branch of a deciduous tree, t-c, ix-xi 2007. exidia glandulosa (bull.): fr. – on a branch of a deciduous tree, t-c, f-u, iii-xi 2007. exidia plana (wiggers) donk. – on dead wood, t-c, x 2007. exidia saccharina (alb. & schwein.) fr. – on wood of pinus sylvestris, t-c xi 2007. flammulina velutipes (m.a. curtis: fr) singer – on deciduous tree stump, t-c, xi-xii 2007. fomes fomentarius (l.: fr.) kickx. – on betula pendula, t-c, f-u, v-xii 2007. fomitopsis pinicola (swart: fr..) p. karst. – on coniferous tree stump, t-c, v-xii 2007. galerina pumila (pers.: fr.) singer – among grass andt mosses, t-c, x 2007. ganoderma applanatum (per.) pat. – on deciduous tree stump, t-c, xi-xii 2007. gymnopus confluens (pers.) p. kumm. – on litter, t-c, f-u, vi-viii 2007. gymnopus dryophilus (bull.) p. kumm. – on litter, t-c, x 2007. gymnopus peronatus (bolt. fr.) antonin, halling & noordel. – on litter, t-c, f-u, vii-x 2007. gyroporus cyanescens (bull.: fr.) quél. – on the ground, t-c, vii 2007, r. hapalopilus rutilans (pers.) p. karst. – on trunks and branches of deciduous tree, t-c, vi 2007. hebeloma mesophaeum (pers.) quél. – on the ground, t-c, x 2007. heterobasidion annosum (fr.) bref. sl. – on pinus sylvestris stump, t-c, xi 2007. hochenbuehelia grisea (peck) singer – on a branch of a deciduous tree, t-c, x 2007. hygrophoropsis aurantiaca (wulf.: fr.) j. schröt. – on the ground, t-c, ix-x 2007. hygrophorus hypothejus var. hypothejus (fr.: fr.) fr. – on the ground in sandy soil, t-c, xi 2007. hymenochaete rubiginosa (schrad.: fr.) lév. – on deciduous tree stump, t-c, vi 2007. hypsizygus ulmarius (bull.: fr.) redhead – on living trunk of ulmus laevis f-u, x-xi 2007, v. inonotus tomentosus (fr.) teng – on the roots of conifers, t-c, vii 2007, v. laccaria amethystina cooke – on the ground, t-c, ix-x 2007. laccaria bicolor (maire) p.d. orton – on the ground, t-c, ix-x 2007. laccaria laccata (scop.: fr.) berk. & broome – on the ground, t-c, x 2007. lactarius camphoratus fr. – on the ground, t-c, viii 2007. lactarius chrysorrheus fr. – on the ground, t-c, x 2007, r. lactarius quietus (fr.) fr. – on the ground, t-c, x 2007. lactarius thejogalus (bull.: fr.) gray ss. neuhoff. – on the ground, t-c, x 2007. lactarius torminosus (schaeft.: fr.) pers. – on the ground, t-c, ix 2007. lactarius vellereus (fr.) fr. – on the ground, t-c, viii 2007. laetiporus sulphureus (bull.: fr.) murrill – on deciduous tree, t-c, viii 2007. leccinum scabrum (bull.: fr.) gray – on the ground, t-c, ix-x 2007. lentinellus cochleatus (pers.: fr.) p. karst. – on deciduous tree stump, t-c, f-u, ix-x 2007. lepiota cristata j.e. lange – on the ground, t-c, ix 2007. lepiota perplexa knudsen – on the ground, t-c, vii 2007. lepista flaccida (sowerby: fr.) pat. – on the ground, t-c, x-xii 2007. lepista gilva (pers.: fr.) pat. – on the ground, t-c, x 2007. lepista nuda (bull.: fr.) cooke. – on the ground, t-c, f-u, ix-x 2007. lycoperdon molle pers. – on the ground, t-c, viii 2007. lycoperdon nigrescens (pers.: pers.) pers. – on the ground, t-c, f-u, ix-x 2007. lycoperdon perlatum (pers.: pers). – on the ground, t-c, f-u, viii-x 2007. macrolepiota procera (scop.: fr.) singer – on the ground, t-c, ix 2007. macrolepiota rhacodes (vittad.) singer – on the ground, t-c, ix-x 2007. marasmius alliaceus (jacq.: fr.) fr. – on the ground, t-c, f-u, ix-x 2007. marasmius rotula (scop.: fr.) fr – on twigs of carpinus betulus, t-c, vii 2007. marasmius scorodonius (fr.: fr.) fr. – on fallen twigs of pinus, t-c, x-xii 2007. melanoleuca schumacheri (fr.: fr.) singer – on the ground, t-c, x 2007. note. species recorded in poland from few sites (wojewoda 2003; flisińska 2004; bujakiewicz 2010). mycena arcangeliana bres. – on decayedtrunks and stumps on deciduous trees, t-c, xi 2007. mycena cinerella (p. karst.) p. karst. – among mosses, t-c, xi 2007. mycena epipterygia ( scop. fr.) gray – on plant debris and fragments of grass, t-c, x 2007. mycena galericulata (scop.: fr.) – dead trunks, t-c, viii 2007. mycena galopus (pers.: fr.) p. kumm. – among mosses, t-c, vii 2007. mycena inclinata (fr.) quél. – on stumps and trunks of deciduous trees, t-c, f-u, vii-xi 2007. 104 a. salamaga and d. ślusarczyk © the author(s) 2014 published by polish botanical society mycena leptocephala (pers.: fr.) millet – in liter, t-c , vii-x 2007. mycena maculata p. karst. – on rotten stumps , t-c, f-u, x-xi 2007. mycena polygramma (bull.: fr.) gray – on stumps of deciduous trees, t-c, x 2007. mycena pura (pers. fr.) p. kumm. – in litter, t-c, f-u, ix-x 2007. mycena purpurefusca (peck) sacc. – in litter, t-c, x 2007. mycena sanguinolenta (alb. & schwein.: fr.) p. kumm. – on litter, t-c, x-xi 2007. mycena stylobates (pers.: fr.) p. kumm. – of fallen leaves of quercus robur, t-c, f-u 2007. mycena viridimarginata p. karst. – in litter, t-c, x 2007. mycena vitilis (fr.) quél. – in litter, t-c, f-u, ix-x 2007. mycena vulgaris (pers.: fr. ) p. kumm. – among mosses, t-c, xi 2007. mycena zephirus (fr. fr.) p. kumm – in litter, t-c, f-u, ix-x 2007. paxillus involutus (batsch.: fr.) fr. s.l. – on the ground, t-c, x 2007. peniophora quercina (pers.: fr.) cooke – fallen branches of quercus robur, t-c, x 2007. phallus impudicus l.: pers. – on the ground, t-c vii-viii 2007. phlebia radiata fr. – on lying trunks of alnus glutinosa, t-c, viii-xi 2007. phlebia tremellosa (schrad.: fr.) nakasone & burds. – on stumps of deciduous tree, xi 2007. phleogena faginea (fr.: fr.) link – on dead of deciduous tree, t-c, xii 2007. note. species recorded in poland in several tens localities (szczepkowski, chachuła 2010; kujawa et al. 2012). pholiota lenta (pers.: fr.) singer – on the ground, t-c, ix-xi 2007. pholiota mutabilis (scop.: fr.) p. kumm. – on stumps of alnus glutinosa, t-c, f-u, x 2007. pholiota squarossa (weigel.: fr.) p. kumm. – on deciduoud wood, t-c, x 2007. piptoporus betulinus (bull.: fr.) p. karst. – on birch wood, t-c, vi-xi 2007. pluteus atricapillus (batsch) fayod – on dead wood, t-c, f-u, x 2007. polyporus brumalis (pers.) fr. – on fallen branches, t-c, f-u, x-xii 2007. psathyrella vernalis velen. – on the ground, among litter, t-c, iv-v 2007. pseudoclitocybe cyathiformis (bull.: fr.) singer – on the ground, t-c, ix-xii 2007. psilocybe aeruginosa (m.a. curtis: fr.) noordel. – on decayed wood, t-c, x 2007. psilocybe fascicularis (huds.: fr.) noordel. – on stumps, t-c, f-u, vii-x 2007. psilocybe lateritia (schaeff.: fr.) noordel. – on stumps of deciduous tree, t-c, f-u, ix-xi 2007. ramaria eumorpha (p. karst.) corner – on the ground, t-c, x 2007. ramaria stricta (pers.: fr.) quél. – on litter, t-c, ix 2007. rhodocollybia butyracea for. asema (fr.fr.) antonin, halling&&noordel. – on litter, t-c, f-u, x 2007. rickenella fibula (bull.: fr.) raith. – among mosses, t-c, vi-viii 2007. ripartites tricholoma (alb. & schwein.: fr.) p. karst. – on the ground, f-u, ix-xii 2007. russula amoenolens romagn. – on the ground, t-c, viii-ix 2007. russula atrorubens quél. – on the ground, t-c, ix 2007. russula emetica (schaeff.) pers.: fr. – on the ground, t-c, f-u, x 2007. russula claroflava grove – on the ground, t-c, x 2007. russula cyanoxantha (schaeff.) fr. – on the ground, t-c, x 2007. russula cyanoxantha var. peltereaui singer – on the ground, t-c, x 2007. russula fragilis var. fragilis (pers.: fr.) – on the ground, t-c, x 2007. russula ochroleuca (pers.) – on the ground, t-c, vii-xi 2007. russula olivascens (pers.) bres. – on the ground, t-c, x 2007. russula paludosa britzelm – on the ground, t-c, x 2007. russula pectinatoides peck – on the ground, t-c, vi 2007. russula solaris ferd. & winge – on the ground, t-c, xi 2007. russula vesca fr. – on the ground, t-c, vii-x 2007. russula viscida kudřna – on the ground, t-c, x 2007. note. (wojewoda 2003; bujakiewicz 2004; szkodzik 2005; miśkiewicz 2000a; gierczyk et al. 2009; karasiński 2009a). russula xerampelina (schaeff.) fr. – on the ground, t-c, viii 2007. schizophyllum commune fr.: fr. – on stumps of deciduous tree, t-c, xii 2007. scleroderma bovista fr. – on the ground, t-c, viii 2007. scleroderma citrinum pers. – on the ground, t-c, vii-ix 2007. scleroderma verrucosum (bull.): pers. – on the ground, t-c, vii 2007. diversity of macromycetes in the białaczów nature reserve 105 © the author(s) 2014 published by polish botanical society sparassis crispa (wulf.): pers. – on the ground, t-c, ix 2007. stereum hirsutum (willd.: fr.) gray – on stumps of deciduous tree, t-c, x-xii 2007. stereum sanguinolentum (alb. & schwein.: fr.) fr. – on branches of coniferous trees, t-c, xi 2007. strobilurus tenacellus (pers.: fr.) singer – on fallen cones of pinus sylvestris, v 2007. suillus luteus (l.: fr.) roussel – on the ground, under pinus sylvestris, t-c, x 2007. trametes gibbosa (pers.: fr.) fr. – on stumps of deciduous tree, t-c, x-xi 2007. trametes versicolor (l.: fr.) pilát – on stumps of deciduous tree, t-c, x-xi 2007. tricholoma sulphureum (bull.: fr.) p. kumm. – on the ground, under quercus robur, t-c x 2007. tubaria furfuracea (pers. fr.) gillet – on fallen twigs, t-c, v 2007. tylopilus felleus (bull.: fr.) p. karst. – on the ground, under pinus sylvestris, t-c, vii 2007. typhula setipes (grev.) berthier – on fallen alnus glutinosa leaves, f-u, f-a, xi 2007. xerocomus badius (fr.: fr.) kühner ex. gilbert – on the ground, t-c, ix-x 2007. xerocomus pascuus (pers.) krombh. – on the ground, t-c, x 2007. xerocomus subtomentosus var. subtomentosus (l.: fr.) quél. – on the ground, t-c, f-u, vii-ix 2007, ix 2008. xeromphalina cornui (quél.) j. favre – on dead wood, t-c, ix 2007. conclusions the undertaken studies in białaczów forest reserve have shown that it is an area of high mycological diversity. species composition of the reserve is influenced by factors like recreation usage of the area, the watercourse in it’s northern part, and the community of tilio-carpinetum calamagrostietosum which is dominat in the reserve and riparian forests: fraxino-alnetum and ficario-ulmetum, growing in smaller amounts. the attempts to evaluate the diversity of macromycetes of the reserve, are interesting, more so because of the wind storm that had stricken the reserve in 2011, almost utterly destroying it. the received data can be the ground to evaluate the fungal changes of białaczów reserve, and used for comparison with mycological studies in future. acknowledgements. we are grateful to the anonymous reviewer for helpful suggestions on the manuscript. the manuscript we dedicate to professor maria ławrynowicz who nearly 50 years ago initiated research on macromycetes in the central poland. references adamczyk j. 2007. grzyby wielkoowocnikowe rezerwatu babsk w województwie łódzkim na tle przemian szaty roślinnej. parki nar. i rez. przyr. 26 (2): 3-16 breitenbach j., kränzlin f.1984. pilze der schweiz i (ascomyceten). mykologia, luzern. breitenbach j., kränzlin f. 1986. pilze der schweiz ii (heterobasidiomycetes, aphyllophorales, gastromycetes). mykologia, luzern. breitenbach j., kränzlin f.1991. pilze der schweiz iii (strobilomycetaceae und boletaceae, paxillaceae, gomphidiaceae, hygrophoraceae, tricholomataceae, polyporaceae). mykologia, luzern. breitenbach j., kränzlin f.1995. pilze der schweiz iv (entolonataceae, pluteaceae, amanitaceae, agaricaceae, coprinaceae, bolbitiaceae, strophariaceae). mykologia, luzern. breitenbach j., kränzlin f. 2000. pilze der schweiz (cortinariaceae). mykologia, luzern. bujakiewicz a. 2004. grzyby wielkoowocnikowe babiogórskiego parku narodowego. 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(eds). 2001. flora agaricina neerlandica. critical monographs on families of agarics and boleti occurring in the netherlands. 5 agaricaceae. a.a. balkema publishers, rotterdam, 169 pp. noordeloos m.e., kuyper t.w., vellinga e.c. (eds). 2005. flora agaricina neerlandica. critical monographs on families of agarics and boleti occurring in the netherlands. 6. taylor & francis group, 226 pp. operat urządzeniowy lasu leśnictwa białaczów 1998. rąkowski g. (ed.). 2006. rezerwaty przyrody w polsce środkowej. instytut ochrony środowiska, warszawa, 168 pp. skirgiełło a. 1960. flora polska. grzyby (mycota) 1: podstawczaki (basidiomycetes), borowikowe (boletales) pwn, warszawa. skirgiełło a. 1991. flora polski. grzyby (mycota) 10: podstawczaki (basidiomycetes), gołąbkowe (russulales), gołąbkowate (russulaceae), gołąbek (russula). pwn, warszawa – kraków. skirgiełło a. 1998. flora polski. grzyby (mycota) 15: podstawczaki (basidiomycetes), gołąbkowe (russulales), gołąbkowate (russulaceae) ii, mleczaj (lactarius). pan, instytut botaniki im. w. szafera, kraków. szczepkowski a., chachuła p. 2010. nowe stanowiska i nowe gatunki żywicieli suchogłówki korowej phleogena faginea (fr.) link w polsce. parki nar. i rez. przyr. 29 (1): 93-121. szkodzik j. 2005. macromycetes in communities of abies alba on its range border in central poland. acta mycol. 40 (1): 113-131. wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland 7. w. szafer institute of botany, polish academy of sciences, kraków, 812 pp. diversity of macromycetes in the białaczów nature reserve 107 © the author(s) 2014 published by polish botanical society wojewoda w., ławrynowicz m. 2006. red list of the macrofungi in poland. (in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. w. szafer institute of botany, polish academy of sciences, kraków: 53-70. różnorodność gatunkowa makromycetes w rezerwacie białaczów (polska środkowa) streszczenie prezentowane dane są wynikiem badań prowadzonych w latach 2006-2007 w rezerwacie białaczów (polska środkowa). celem badań było poznanie składu gatunkowego grzybów makroskopowych i przedstawienie ich na tle drzewostanu i siedliska. łącznie zidentyfikowano 167 taksonów grzybów makroskopowych (13 gatunków ascomycota i 154 basidiomycota). w rezerwacie, dominują grzyby saprotroficzne – 118 gatunków, grzybów mikoryzowych zebrano 44 gatunki, a pasożytniczych 5 gatunków. pod względem ekologicznym przeważają grzyby tworzące owocniki na ziemi (74) oraz na drewnie (62), natomiast grzyby naściółkowe reprezentowane są przez 31 gatunków. w rezerwacie odnotowano także jedenaście gatunków z czerwonej listy grzybów wielkoowocnikowych zagrożonych w polsce: phleogena faginea (e), hypsizygus ulmarius, mycena purpureofusca, inonotus tomentosus (v), clavariadelphus fistulosus, c. junceus, gyroporus cyanescens, lactarius chrysorrheus, pachyphloeus melanoxanthus (r), russula amoenoleus, hygrophorus hypothejus var. hypothejus (i). przedstawione wyniki z jednej strony stanowią dokumentację mykologiczną, a z drugiej materiał wyjściowy do badań porównawczych w procesie zmian w następstwie zniszczenia rezerwatu przez wichurę w 2011 roku. 2014-06-30t22:57:33+0200 polish botanical society 2014-08-20t21:45:24+0200 polish botanical society 2014-08-20t22:46:44+0200 polish botanical society 2014-08-25t21:39:32+0200 polish botanical society cladonia metacorallifera, a lichen species new to the eastern carpathians piotr osyczka1 and robert kościelniak2 1department of polar research and documentation, institute of botany jagiellonian university, kopernika 27, pl-31-501 kraków, piotr.osyczka@uj.edu.pl 2department of botany, institute of biology, pedagogical university of kraków, podbrzezie 3 pl-31-054 kraków osyczka p., kościelniak r.: cladonia metacorallifera, a lichen species new to the eastern carpathians. acta mycol. 44 (2): 233–238, 2009. a rare lichen cladonia metacorallifera was found in the bieszczady mts. and this is the first report of the species from the eastern carpathians. in poland, its occurrence is probably limited to small populations scattered in high mountain situations. the data of distribution of the species in poland, ecological characterization of the new locality and some taxonomical remarks concerning the recorded specimen, as well as a photograph of its habit are provided. key words: lichens, section cocciferae, distribution, ecology, bieszczady mts. introduction cladonia metacorallifera asahina is a fruticose, cup-shaped and red-fruited member of the section cocciferae (delise) a. evans (fig. 1). the huge variability of the morphology of the species in this group causes many problems in their taxonomy and identification (e.g., stenroos 1989a; osyczka 2009). generally, the podetia of c. meta corallifera are throughout densely covered by squamules, microsquamules and/ or cortical granules, and furthermore decorticated parts are often blackened, especially towards the tips of podetia. the thallus contains usnic, didymic and squamatic (or thamnolic) acids and this chemical feature is diagnostically important because it differentiates the species from morphologically similar cup-shaped cladonia from the section (e.g., c. coccifera (l.) willd., c. diversa asperges and c. borealis s. stenroos). the significant morphological and chemical differences between c. metacorallifera and other scyphose lichens with red apothecia are discussed in several papers (e.g., stenroos 1989b; see also kowalewska, kukwa 2004; osyczka et al. 2006). acta mycologica vol. 44 (2): 233–238 2009 dedicated to professor krystyna czyżewska in honour of 40 years of her scientific activity 234 p. osyczka and r. kościelniak the bieszczady mts. are an important centre of lichen biodiversity in poland (e.g., czyżewska 2003; kościelniak 2007, 2008). the intensive lichenological field research in this region carried out in the last decade still provides new information about the occurrence and distribution of lichens in poland and the carpathians. during a critical revision of lichen materials belonging to cladonia coccifera s.l. deposited in herbarium krap-l, one specimen from the bieszczady national park turned out to be cladonia metacorallifera. this is the first report on the taxon in this mountain range, as well as in the whole area of the eastern carpathians. fig. 1. specimen of cladonia metacorallifera from the bieszczady mts. (leg. j. kiszka, r. koscielniak, 13 july 2005, krap-l). scale bar = 1 cm. cladonia metacorallifera 235 material and methods the revision of cladonia coccifera s.l. specimens housed in krap-l (pedagogical university of kraków) was done using standard microscopic techniques supported by chemical analysis. lichen substances were determined by tlc in solvent a and c (acc. to orange et al. 2001). metter-toledo seveneasy ph meter was employed for substratum ph determination. the analysis was made by suspending substrate matter in pure water. all known polish localities of cladonia metacorallifera are mapped according to the atpol grid square system (zając 1978; modified by cieśliński, fałtynowicz 1993). the grid square 1×1 km was used for showing the precise locality in the bieszczady national park. results and discussion cladonia metacorallifera asahina, j. jap. bot. 15: 612 (1939). it is a new component of the epigeic biota of mountain pasture lichens in the bieszczady mts. (fig. 2). it was recorded only on w slope of rozsypaniec massif, fig. 2. locality of cladonia metacorallifera in the bieszczady mts. cladonia metacorallifera 237 also include one situation in the karkonosze mts. (kowalewska, kukwa 2004 – fig. 3). a taxonomical revision of cladonia coccifera s.l. deposited in polish herbaria did not provide any further c. metacorallifera localities. it seems that in poland the lichen is a rare species and its occurrence is limited to small populations scattered in the highest mountainous areas along the southern country border. acknowledgements. a part of this research was supported by the ministry of science and higher education (grant no n n305 2012 35). we would like to thank an anonymous reviewer for helpful remarks and corrections of the manuscript. references asahina y. 1939. japanische arten der cocciferae (cladonia-coenomyce). j. jap. bot. 15: 602–620. cieśliński s., fałtynowicz w. (eds). 1993. atlas of the geographical distribution of lichens in poland. 1. w. szafer institute of botany, polish academy of sciences, kraków. czyżewska k. 2003. evaluations of the threat to lichens in poland. monogr. bot. 91: 241–249. kowalewska a., kukwa m. 2004. cladonia metacorallifera (lichenized ascomycota, cladoniaceae) new to poland and additional record from slovakia. biologia, bratislava 59 (4): 433–434. kościelniak r. 2007. usnea florida – threatened species of rich biotopes in the polish eastern carpathians. acta mycol. 42 (2): 281–286. kościelniak r. 2008. the importance of primeval and natural forests to preservation of species diversity of lichens in the bieszczady mts. roczniki bieszczadzkie 16: 67–76. orange a., james p.w., white f. j. 2001. microchemical methods for identification of lichens. british lichen society, london, 101 pp. osyczka p. 2004. cladonia macroceras (delise) hav. (in:) u. bielczyk, s. cieśliński, w. fałtynowicz (eds). atlas of geographical distribution of lichens in poland. 4. w. szafer institute of botany, polish academy of sciences, kraków: 29–33. osyczka p. 2006. the lichen genus cladonia (cladoniaceae, lichenized ascomycota) from spitsbergen. pol. polar res. 27 (3): 207–242. osyczka p. 2009. cladonia diversa (cladoniaceae, lichenized ascomycota) – overlooked lichen in poland. acta. soc. bot. pol. (in print). osyczka p., węgrzyn m., flakus a. 2006. two species of the genus cladonia (cladoniaceae, lichenized, ascomycota) new to the polish tatra mts. polish bot. j. 51 (2): 233–235. osyczka p., flakus a., węgrzyn m., cykowska b. 2007. cladonia crispata var. cetrariiformis (cladoniaceae, lichenized ascomycota) in the tatra mts. biologia, bratislava 62 (2): 144–147. pišút i. 1997. interessantere flechtenfunde aus der slowakei 4. bull. slov. bot. spoločn., bratislava 19: 68–71. stenroos s. 1989a. taxonomy of the cladonia coccifera group. 1. ann. bot. fennici 26: 157–168. stenroos s. 1989b. taxonomy of the cladonia coccifera group. 2. ann. bot. fennici 26: 307–317. wirth v. 1995. die flechten baden-württembergs. 1. verlag e. ulmer, stuttgart, 527 pp. zając a. 1978. atlas of distribution of vascular plants in poland (atpol). taxon 27: 481–484. 238 p. osyczka and r. kościelniak cladonia metacorallifera, nowy porost dla karpat wschodnich streszczenie cladonia metacorallifera asahina jest kieliszkowato zakończonym porostem należącym do sekcji cocciferae (delise) a. evans. duża zmienność morfologiczna taksonów z tej sekcji wciąż stwarza problemy taksonomiczne i utrudnia prawidłową identyfikację gatunków (por. stenroos 1989a; osyczka 2009). cechą charakterystyczną c. metacorallifera są liczne łuseczki na ogół gęsto rozmieszczone na całej długości podecjów oraz czerniejące fragmenty plechy widoczne w miejscach pozbawionych kory i łuseczek. cladonia metacorallifera wytwarza kwas usninowy, didymowy oraz skwamatowy (lub tamnoliowy). ta właściwość chemiczna jest ważną cechą diagnostyczną gatunku. istotne cechy taksonomiczne odróżniające c. metacorallifera od morfologicznie podobnych taksonów z sekcji cocciferae (np. c. coccifera (l.) willd., c. diversa asperges lub c. borealis s. stenroos) zawarte zostały w kilku publikacjach (np. stenroos 1989b; zobacz również: kowalewska, kukwa 2004; osyczka et al. 2006). w polsce po raz pierwszy c. metacorallifera znaleziono na jednym stanowisku w masywie śnieżki w karkonoszach (kowalewska, kukwa 2004), następnie podano go z dwóch stanowisk w polskiej części tatr wysokich (osyczka et al. 2006). w wyniku rewizji materiałów zielnikowych porostów należących do c. coccifera s.l., zdeponowanych w zielniku lichenologicznym uniwersytetu pedagogicznego w krakowie (krap-l), stwierdzono występowanie c. metacorallifera również w polskiej części bieszczadów. jest to pierwsze doniesienie o występowaniu tego gatunku w karpatach wschodnich. nowe stanowisko c. metacorallifera znajduje się w granicach bieszczadzkiego parku narodowego, na zachodnim zboczu masywu rozsypańca, na wysokości 1153 m n.p.m., ponad górną granicą lasu, w obrębie rumoszu skalnego. stanowisko to charakteryzuje się wysokim nasłonecznieniem i ekspozycją południowo-zachodnią. porost ten rósł wśród mchu na humusie (ph = 4.65). występowanie c. metacorallifera w polsce prawdopodobnie ograniczone jest do niewielkich populacji rozproszonych w obszarach wysokogórskich wzdłuż południowej granicy kraju. 2014-01-01t11:49:39+0100 polish botanical society 2014-08-29t18:51:02+0200 polish botanical society 2014-08-26t20:49:32+0200 polish botanical society 2014-08-27t16:49:25+0200 polish botanical society in memoriam doctor habilitus vincentas kazys urbonas (1934 – 2008) on february 3, 2008, the distinguished lithuanian mycologist, vincentas kazys urbonas passed away at his home in vilnius. he was well known researcher on macroscopic fungi, and a committed person in nature conservation, science popularization and administration. vincentas urbonas was born in gudeliai village (marijampolė district, southern lithuania) on june 27, 1934. after graduation from the secondary school in simnas he entered the faculty of nature and geography of vilnius pedagogical institute (now – vilnius pedagogical university), and completed the studies of biology in 1959.after graduation v. urbonas started the scientific work at the institute of botany of the lithuanian academy of sciences in vilnius. his main field of research was the diversity, taxonomy, biology and ecology of agaricoid fungi. in 1967 v. urbonas received a degree of candidate of biological sciences, based on a dissertation “naujas indėlis lietuvos tsr kepurėtiesiems (agaricales eilės) grybams pažinti” [new contribution to the knowledge of the agaricales s.l. of lithuanian ssr] under the supervision of dr. jonas mazelaitis. in 1990 he defended his doctor of science thesis in biology “agarikal’nye griby (agaricales s.l.) litovskoj ssr (vidovoj sostav, sistematika, ekologija)” [the agaricales s.l. of lithuanian ssr (species composition, systematics, ecology)] at m.g. kholodny institute of botany of the national academy of sciences of ukraine in kiev. after nostrification in 1994 he was bestowed a scientific title of doctor habilitus. during his scientific career v. urbonas published more than 110 scientific papers and 7 monographs. he had wide-ranging interests in mycology. most of his works were devoted to the diversity and distribution of lithuanian agaricoid fungi. more than ten thousand fungus specimens collected and determined by v. urbonas make basis for the agaricoid collection of the herbarium of the institute of botany (bilas). about 1260 species of agaricoid fungi recorded in lithuania were described and illustrated in five volumes of edition “lietuvos grybai – mycota lithuaniae“. in co-operation with prof. k. kalamees and dr. v. lukin two editions of “conspectus florum agaricalium fungorum (agaricales s.l.) lithuaniae, latviae et estoniae” were published. v. urbonas proposed several taxonomic novelties (new suprageneric acta mycologica vol. 43 (2): 231–236 2008 232 j. kasparavičius and e. kutorga taxa, nomenclatural combinations). v. urbonas investigated mycobiota in protected areas, studied rare and endangered species of fungi. he dealt with problems of fungi conservation, and was actively involved in making lists of fungi for the lithuanian red data book. together with colleagues he investigated biochemical properties, heavy metals and radionuclides in the fruit bodies of the agaricoid fungi. he was also interested in resources of edible mushrooms and cultivation of some mushroom species, especially agaricus bisporus. under his supervision several active lithuanian mycologists, namely antanas matelis, danutė stankevičienė and ernestas kutorga received cand. biol. degree, and jonas kasparavičius received ph. d. degree. as a consultant, a member of defence committees or as an official opponent he took part in ph.d. student research and thesis defence process of many mycologists and botanists. v. urbonas attended numerous local and international scientific conferences. he took part in almost every symposium of baltic mycologists and lichenologists and, when it was lithuanian turn to organize the event, was often a heart and soul of the organizing committees. in 1990ties when the independence of lithuania was restored, v. urbonas took initiative to renew and activate the scientific contacts between lithuanian and polish mycologists and lichenologists. this initiative resulted in bilateral visits of researches and exchange of mycological literature published in both countries. v. urbonas was active and successful administrator. from 1984 to 2001 he was a head of the laboratory of cryptogamous plants (since 1992 – laboratory of mycology), for some period served as a deputy director and a member of the council of institute of botany. he was one of initiators, at first a deputy editor-in-chief and later editor-in-chief of the many-volume edition “lietuvos grybai – mycota lithuaniae“. also he was a member of editorial board of scientific journal “botanica lithuanica“ and a member of botanical dictionary commission. vincentas urbonas worked very effectively for popularisation of mycology. he organised or actively participated in fungal exhibitions, delivered public lectures, gave interviews on tv and radio, wrote numerous popular papers. he was the author or co-author of popular books: “lietuvos grybai” [mushrooms of lithuania] (1980, together with j. mazelaitis), “grybai” [mushrooms]” (1986, 1998), “mažieji miško turtai” [lesser riches of forest] (1987, together with co-authors), “susipažinkime: grybai” [get acquainted: mushrooms] (1998), and “lietuvos grybų atlasas” [atlas of lithuanian mushrooms] (2007). he was active member of the lithuanian mycological society since its foundation in 1998. dr. habil. v. urbonas scientific work was acknowledged by the lithuanian state award for science in 1981 (together with j. mazelaitis and a. minkevičius) for the cycle of works “lietuvos tsr makromicetai” [macromycetes of lithuanian ssr] and the lithuanian state award for science in 2002 (together with m. ignatavičiūtė, s. stanevičienė, b. grigaliūnaitė and e. kutorga) for the cycle of works “lietuvos makromicetų ir augalų parazitinių mikromicetų įvairovės įvertinimas (1991–2000)” [evaluation of diversity of macromycetes and plant pathogenic micromycetes of lithuania (1991–2000)]. he was a devoted family man. with his wife jūratė they raised son raimundas and daughter živilė, also took care of beloved grandchildren. we remember him as a skilled scientist, talented writer, and patient teacher, warm and considerate colleague always ready to assist. we will miss his kind words and encouragement in which he was always so generous. bibliography of publications 233 i. monographs urbonas v., kalamees k., lukin v. 1974. konspekt izuchenija agarikovykh gribov litovskoj ssr, latvijskoj ssr, estonskoj ssr. [synopsis of agaricoid fungi of lithuanian ssr, latvian ssr, estonian ssr]. mintis, vilnius, pp. 129. (in russian). urbonas v., kalamees k., lukin v. 1986. conspectus florum agaricalium fungorum (agaricales s.l.) lithuaniae, latviae et estoniae. mokslas, vilnius, pp. 139. (in latin). urbonas v. 1997. lietuvos grybai (mycota lithuaniae), 8 (1). kempiniečiai (polyporales), žvynbaravykiečiai (strobilomycetales), baravykiečiai (boletales), guoteniečiai (hygrophorales). valstiečių laikraštis, vilnius. pp. 200. (in lithuanian). urbonas v. 1997. lietuvos grybai (mycota lithuaniae), 8 (2). baltikiečiai (tricholomatales). valstiečių laikraštis, vilnius, pp. 216. (in lithuanian). urbonas v. 1999. lietuvos grybai (mycota lithuaniae), 8 (3). agarikiečiai (agaricales), gijabudiečiai (entolomatales). valstiečių laikraštis, vilnius, pp. 296. (in lithuanian). urbonas v. 2001. lietuvos grybai (mycota lithuaniae), 8 (4). musmiriečiai (amanitales), ūmėdiečiai (russulales). valstiečių laikraštis, vilnius, pp. 224. (in lithuanian). urbonas v. 2005. lietuvos grybai (mycota lithuaniae), 8 (5). nuosėdiečiai (cortinariales). valstiečių laikraštis, vilnius, pp. 288. (in lithuanian). ii. main research papers mazelaitis j., gricius a., urbonas v. 1963. naujos lietuvos tsr florai buožiagrybių (basidiomycetes) rūšys. [new for lithuanian ssr species of basidiomycetes]. lietuvos tsr ma darbai, ser. c, 32 (3): 89-101. (in lithuanian). urbonas v. 1966. nekotorye dannye k materialu flory bazidiomicetov litovskoj ssr. [some data on flora of basidiomycetes of lithuanian ssr]. lietuvos tsr ma darbai, ser. c, 41 (3): 59-68. (in russian). urbonas v. 1966. novyje vidy bazidial‘nykh gribov v litovskoj ssr. [new basidiomycete species for lithuanian ssr]. lietuvos tsr ma darbai, ser. c, 39 (1): 23-28. (in russian). urbonas v. 1966. obzor gribov semeistva boletaceae litovskoj ssr. [survey of fungi from the family boletaceae in lithuanian ssr]. (in:) uchenye zapiski latvijskogo gosudarstvennogo universiteta imeni petra stuchki, botanika 74 (2): 134-140. (in russian). urbonas v. 1970. 22 novykh i 4 redkikh dlia flory agarikovykh gribov litovskoj ssr vida. [22 new and 4 rare species for flora of agaricoid fungi of lithuanian ssr]. lietuvos tsr ma darbai, ser. c, 52 (2): 55-64. (in russian). urbonas v. 1970. 41 novyj dlia flory agarikovykh gribov litovskoj ssr vid. [41 species new for flora of agaricoid fungi of lithuanian ssr]. lietuvos tsr ma darbai, ser. c, 52 (2): 41-54. (in russian). urbonas v. 1972. novye dannye o stropharia albocrenulata (pk.) kreisel. [new data on stropharia albocrenulata]. mikol. fitopatol. 6 (1): 74-77. (in russian). urbonas v. 1973. k sistematike i rasprostraneniji gribov semejstva strophariaceae sing. et smith v sssr (1. rod stropharia (fr.) quél). [notes on systematics and distribution of fungi from the family strophariaceae in ussr (1. the genus stropharia)]. lietuvos tsr ma darbai, ser. c, 62(2): 9-24. (in russian). urbonas v. 1973. materialy k flore i ekologii agarikovykh gribov litovskoj ssr (2. novye redkie nakhodki rozovosporovykh gribov). [materials on flora and ecology of agaricoid fungi of lithuanian ssr (2. new rare records of fungi with rose-coloured spores)]. lietuvos tsr ma darbai, ser. c, 66 (2): 21-29. (in russian). urbonas v. 1973. materialy k flore i ekologii agarikovykh gribov litovskoj ssr (3. novye vidy belosporovykh gribov). [materials on flora and ecology of agaricoid fungi of lithuanian ssr (3. new species of fungi with hyaline spores)]. lietuvos tsr ma darbai, ser. c, 67 (3): 21-25. (in russian). urbonas v. 1973. materialy k flore i ekologii agarikovykh gribov litovskoj ssr (4. novye vidy gribov rodov lepiota i mycena). [materials on flora and ecology of agaricoid fungi of lithuanian ssr (4. new species of the genera lepiota and mycena)]. lietuvos tsr ma darbai, ser. c, 67 (3): 13-19. (in russian). urbonas v. 1973. materialy k flore i ekologii agarikovykh gribov litovskoj ssr (5. novye vidy gribov rodov agrocybe, conocybe, inocybe). [materials on flora and ecology of agaricoid fungi of lithuanian ssr (5. new species of the genera agrocybe, conocybe, inocybe)]. lietuvos tsr ma darbai, ser. c, 70 (3): 15-19. (in russian). 234 j. kasparavičius and e. kutorga urbonas v. 1973. materialy k flore i ekologii agarikovykh gribov litovskoj ssr (6. novye vidy burosporovykh gribov). [materials on flora and ecology of agaricoid fungi of lithuanian ssr (6. new species of fungi with brown spores)]. lietuvos tsr ma darbai, ser. c, 83 (3): 9-24. (in russian). urbonas v. 1973. novye vidy agarikovykh gribov dlja sovetskogo sojuza. [new species of agaricoid fungi for the soviet union]. lietuvos tsr ma darbai, ser. c, 55 (25): 15-18. (in russian). urbonas v. 1974. materialy k flore i ekologii agarikovykh gribov litovskoj ssr (1. novyje vidy gribov roda rhodophyllus quél.). [materials on flora and ecology of agaricoid fungi of lithuanian ssr (1. new species of the genus rhodophyllus)]. lietuvos tsr ma darbai, ser. c, 66 (2): 21-29. (in russian). urbonas v. 1975. k sistematike i rasprostraneniju gribov semejstva strophariaceae sing. et smith v sssr (2. rod hypholoma (fr.) kummer). [notes on systematics and distribution of fungi from the family strophariaceae in ussr (2. the genus hypholoma)]. lietuvos tsr ma darbai, ser. c, 72 (4): 3-18. (in russian). urbonas v. 1978. k sistematike i rasprostraneniju gribov semejstva strophariaceae sing. et smith v sssr (3. rod psilocybe (fr.) kummer). [notes on systematics and distribution of fungi from the family strophariaceae in ussr (3. the genus psilocybe)]. lietuvos tsr ma darbai, ser. c, 81 (1): 9-18. (in russian). dušauskienė-duž r., urbonas v. 1978. estestvennaja radioaktivnost’ nekotorykh agarikovykh gribov, obuslovlennaja svincom -210. [natural radioactivity of some agaricoid fungi, determined by pb210]. mikol. fitopatol. 12 (4): 280-284. (in russian). dušauskienė-duž r., urbonas v. 1978. o migracii pb210 i sr90 v nazemnykh biogeocenozakh. [data on migration of pb210 and sr90 in land biogeocenoses]. ekologija 5: 97-100. (in russian). mazelaitis j., urbonas v. 1981. redkij i maloizvestnyj vid roda coprinus (fr.) s. f. gray. (opisanie shljapochnogo griba, obnaruzhennogo v palange). 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(in lithuanian). jonas kasparavičius and ernestas kutorga 2014-01-01t11:48:24+0100 polish botanical society wentiomyces sp. from plant litter on poor fen in northeastern poland 237this is an open access article distributed under the terms of the creative commons attribution 3.0 license (creativecommons.org/licenses/by/3.0/), which permits redistribution, commercial and non-commercial, provided that the article is properly cited. © the author(s) 2014 published by polish botanical society acta mycologica introduction the results of the extensive literature survey published by thormann and rice [1] have shown that more than 600 species of fungi were already recorded from various types of peatland habitats. peatlands are one of the major carbon-sequestrating ecosystems (e.g. [2]), and the apparent negative influence of various anthropogenic disturbances, e.g. global warming and water-level drawdown on decomposition processes, has stimulated research on many interconnected ecological aspects, including interactions with microbial decomposers, and especially fungi (e.g. [3]). original research paper acta mycol 49(2):237–247 doi: 10.5586/am.2014.021 received: 2014-04-15 accepted: 2014-10-14 published electronically: 2014-12-01 wentiomyces sp. from plant litter on poor fen in northeastern poland mateusz wilk1,2*, julia pawłowska3, marta wrzosek3 1 department of plant ecology and environmental protection, faculty of biology, university of warsaw, al. ujazdowskie 4, 00-478 warsaw, poland 2 college of inter-faculty individual studies in mathematics and natural sciences, university of warsaw, żwirki i wigury 93, 00-089 warsaw, poland 3 department of plant systematics and geography, faculty of biology, university of warsaw, al. ujazdowskie 4, 00-478 warsaw, poland abstract during research carried on peatlands in northeastern poland, small leaf-litter ascomycete identified as wentiomyces sp. was found on decaying litter of sphagnum sp. and vaccinium oxycoccos. the taxonomy of this genus remains obscure, and there are still few and incidental published records in the mycological literature. this taxon is therefore described in the present paper in more detail and a short overview on the current knowledge regarding ecology and systematics of wentiomyces spp. is provided. keywords: leaf-litter ascomycetes; peatlands; pseudoperisporiaceae; sphagnum; vaccinium oxycoccos; oxycoccus palustris * corresponding author. email: mwilk@student.uw.edu.pl handling editor: maria rudawska http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.2014.021 mailto:mwilk%40student.uw.edu.pl?subject=am.2014.016 238© the author(s) 2014 published by polish botanical society acta mycol 49(2):237–247 wilk et al. / wentiomyces sp. from plant litter fungi are described to be the most important group of decomposers in these ecosystems [4,5], and growing interest regarding different aspects of fungal diversity and role on peatlands can be observed (e.g. [6,7]). however, still surprisingly few of those studies concern the diversity and potential role of the leaf-litter microfungal communities (e.g. [8]). our knowledge and understanding of the factors involved in the decomposition processes on peatlands and the possible interactions of these with climate change and anthropogenic disturbances are therefore incomplete without more research on peatland mycobiota. during our research on leaf-litter microfungi from peatlands, interesting specimens were found, which were identified as wentiomyces sp. the concept of this genus is still little understood, therefore the aim of this paper was to provide some details concerning our findings and review the current knowledge regarding the taxonomy and biology of the genus wentiomyces. material and methods findings described in this paper originate from a research project carried in 2010–2014 on peatlands in the szeroki bór mire complex, near town of pisz, the warmińskomazurskie voivodeship, northeastern poland. the study site, where the presented fungal specimens were found, was located on open poor fen, being in fact a small lake completely overgrown by dense cover of sphagnum spp. with carex limosa l., eriophorum vaginatum l., vaccinium oxycoccos l. and drosera rotundifolia l.; dead pine trees occur on the banks. the material collected in october 2010 for preliminary screening, comprised of mixed litter of moribund and healthy sphagnum spp. fragments with addition of dead leaves and stems of vaccinium oxycoccos. the method of incubation of plant litter samples in damp chambers was employed for the study [9]. material was observed under dissecting microscope smz800, and semipermanent prep-slides were made in lactophenol-blue or glycerin, and observed under nikon eclipse e200 light microscope. all prep-slides and photographs are housed in the mycological laboratory of the department of plant systematics and geography, faculty of biology, the university of warsaw. taxonomic system and nomenclature for fungi follow mycobank [10], and for plants follow integrated taxonomic information system [11] except for the table, where they are given as in original publications. descriptions of specimens wentiomyces koorders, pseudoperisporiaceae, dothideomycetidae, dothideomycetes. wentiomyces sp., specimen a (fig. 1a). superficial spherical pseudothecium, ca. 42 μm in diameter, brown; peridium composed of polyhedral cells, often 4–6 μm in diameter; around conspicuous ostiole few characteristic, rigid, dark brown and thick walled setae, ca. 10–11 μm long, 3–4 μm wide, mostly twice dichotomously branched and blunt at the apices. setae have one septum, and are paler to hyaline towards the tips. pseudothecium grows on sparse, superficial, melanized mycelium; on the bottom part of ascomata melanized, thread-like hyphae emerge, ca. 2–5 μm in diameter. no asci or ascospores observed. habitat. on moribund sphagnum spp. leaves collected in october 2010; poor fen in szeroki bór peatland complex, 53°36'20.77"n, 21°37'56.79"e. first record from this host. 239© the author(s) 2014 published by polish botanical society acta mycol 49(2):237–247 wilk et al. / wentiomyces sp. from plant litter wentiomyces sp., specimen b (fig. 1b). pseudothecium superficial, spherical, ca. 50 μm in diameter, brown, peridium composed of polyhedral cells. ostiole conspicuous, surrounded by eight characteristic, rigid, dark brown and thick walled setae, ca. 11–14 μm long, 4–5 μm wide, with one septum, at the apex mostly twice dichotomously branched, paler to hyaline towards the tips. on the bottom part of the ascomata, melanized threadlike hyphae emerge, ca. 2–5 μm in diameter. no asci or ascospores observed. habitat. on dead leaf of vaccinium oxycoccos collected in october 2010; szeroki bór peatland complex, 53°36'20.77"n, 21°37'56.79"e. first published record from this host. discussion despite of more than a century since its discovery, the taxonomy of the genus wentiomyces remains extremely complicated. it was raised by koorders [12], to encompass a single species, w. javanicus koord., characterized by superficial, hypophyllous mycelium with superficial pseudothecia (perithecia in original description) possessing in the upper part characteristic, melanized, dichotomously branched at the apex setae, and on the lower part of the ascomata brownish, long hair-like hyphal appendages (although called setae in the original description); asci clavate with eight ascospores; ascospores hyaline, with median septa; no paraphyses. koorders himself mentions, however, that his material is sparse, and mostly immature, and that the color of mature ascospores could potentially change to brownish, although he states this was unlikely [12]. the original herbarium material of the type is unavailable [13,14], and this is the cause for several taxonomical rearrangements of this genus, of which none can still be considered final, because epitypification was never conducted. hansford [15] first mentioned the possible synonymy of the wentiomyces and dimeriella speg., but it was müller and arx [13], who finally concluded that these two genera are in fact synonymous, and have given the name wentiomyces priority over dimeriella, and eleven other generic names fig. 1 specimens of wentiomyces sp. collected on poor fen. pseudothecium with characteristic dichotomously lobed setae around ostiole; specimen from sphagnum sp. (a). specimen from vaccinium oxycoccos (b). scale bars: 20 μm. 240© the author(s) 2014 published by polish botanical society acta mycol 49(2):237–247 wilk et al. / wentiomyces sp. from plant litter considered synonymous. it has to be noted, that in their understanding of the genus, the shape of the setae was no longer considered specific, therefore wentiomyces fide müller and arx [13] included also species with simple, acute or blunt setae; in addition some taxa included have sparse paraphysoids. farr [16] however claimed wentiomyces to be a doubtful genus, which should not be used in favor of dimeriella. this was further applied and expanded by barr [17], who rejected wentiomyces, and transferred six species (including two subspecies) with dichotomously branched setae to neocoleroa petrak, and one lichenicolous species with straight, acute setae to raciborskiomyces siemaszko. to further complicate the situation, this rearrangement is not widely and uniformly accepted. in addition, etayo and sancho [18], transferred two lichenicolous taxa with straight setae to niesslia auersw. because of this chaos, it is even difficult to estimate the correct number of the species included in wentiomyces. after their treatment from 1962 [13], arx and müller [19] concluded that wentiomyces contains “about 50 species”, the large number being of course due to synonimization of dimeriella with wentiomyces, and this was later adopted by kirk et al. [20]. search in index fungorum database [21], however, brings only 21 records, with 20 species and two subspecies. of these, wentiomyces fuliginosus (woron.) e. müll. is treated as a synonym of antenulariella fuliginosa woron., wentiomyces hirtulus (speg.) e. müll. as a synonym of dimeriella hirtula speg., and wentiomyces peltigericola d. hawksw. as a synonym of niesslia peltigericola (d. hawksw.) etayo. search in the mycobank database [10] brings the same number of results, but without clear statements on the status of the abovementioned three species, suggesting 20 existing species for wentiomyces. that is due to the fact, that no formal combinations were created for the vast majority of dimeriella species to be included in wentiomyces, even in the abovementioned studies by müller and arx [13], and arx and müller [19]. this situation clearly indicates that the genus wentiomyces is in urgent need for revision, and the chaos here is corroborated by the fact, that there are no molecular sequences of any of the described species [22]. it is also worth noting, that the names wentiomyces and dimeriella are on the preliminary draft of the list of protected generic names for fungi [23], and therefore the taxonomic and phylogenetic relationships between and within them will have to be resolved. if the mycobank classification system is adopted, the genus wentiomyces comprises species, which grow as saprotrophs and/or biotrophs on plants, and as specialized lichenicolous fungi (tab. 1). most plant-associated taxa grow hypophyllously, with only few recorded also from plant stems. some authors label wentiomyces spp. as pathogens, causing leaf spot diseases ([24] for w. javanicus). it appears, that some taxa can also penetrate leaf cuticle [e.g. w. clavisetus (doidge) arx] [15]; in case of lichenicolous taxa, hansford [25] mentions that w. lichenicola (hansf.) d. hawksw. penetrates the thallus of a lichen. lichenicolous species occur on terrestrial lichens (w. peltigericola on peltigera spp.), but also on epiphyllous lichens (w. lichenicola on fellhanera and bacidina). according to the usda fungus-host database [26], there are 38 published reports on 14 species of wentiomyces from 16 countries; half of the reports are from tropical countries; these are however incomplete data, because many other reports, especially for lichenicolous species, exist (tab. 1). there is a recent finding of wentiomyces sp. from poland [27] on dead leaves of rhododendron tomentosum harmaja from poor fen. we know also on the findings on other ericaceae from raised bog in siberia (n.v. filippova, personal 241© the author(s) 2014 published by polish botanical society acta mycol 49(2):237–247 wilk et al. / wentiomyces sp. from plant litter sp ec ie s su bs tr at um lo ca lit y r ef er en ce s w en tio m yc es a lp iv ag us n og ra se k sy n. n eo co le ro a al pi va ga (n og ra se k) m .e . b ar r si le ne a ca ul is (a ); c ar ex fi rm a (b ); sa xi fr ag a ca es ia (c ); sa xi fr ag a pa ni cu la ta (d ) (a ) s w ed en ; ( b –d ) a us tr ia (a –d ) [ 32 ] w . c la vi se tu s ( d oi dg e) a rx sy n. d im er ie lla c la vi se ta d oi dg e; c ol er oa cl av is et a (d oi dg e) h an sf . a st er ac ea e, e .g . v er no ni a an ul ifo lia (a ); er la ng ea m ar gi na ta (b ); c or di a sp . ( c ); va rr on ia s p. (d ); m ik an ia s p. (e ) (a ) u ga nd a; (b ) m al aw i; (c ,d ) w es t i nd ie s, c os ta r ic a, n or th er n so ut h a m er ic a; (e ) e as t an d so ut h a fr ic a, g ha na (a ) [ 13 ,1 5] ; ( b ) [ 26 ]; (c –e ) [ 33 ] w . c lu si ae j. l. b ez er ra & p or oc a le av es o f c lu si a m el ch io ri v en ez ue la , b ra zi l [2 6, 34 ] w . d ry ad is k . h ol m & l . h ol m sy n. n eo co le ro a dr ya di s ( k . h ol m & l . h ol m ) m .e . b ar r d ea d le av es o f d ry as o ct op et al a (a ); c as si op e te tr ag on a (b ) (a ,b ) n or w ay (s va lb ar d) [3 5] w . fi m br ia tu s ( d ea rn . & h ou se ) m .e . b ar r sy n. v en tu ri a fim br ia ta d ea rn . & h ou se ; la si os te m m a fim br ia tu m (d ea rn . & h ou se ) m .e . b ar r; n eo co le ro a fim br ia ta (d ea rn . & h ou se ) m .e . b ar r o n la ng ui sh in g le av es o f a nt en na ri a sp . ( a ); a ch ill ea m ill ef ol iu m (b ); fi lip en du la u lm ar ia (c ); sa xi fr ag a ca es pi to sa (d ); si le ne a ca ul is (e ) (a ) u sa ; ( b, d ,e ) c an ad a; (c ) s w ed en (a ) [ 36 ]; (c ) [ 26 ]; (b ,d ,e ) [ 37 ,3 8] w . f ul ig in os us (w or on .) e. m ül l. sy n. a nt en nu la ri el la fu lig in os a w or on . li ve le av es o f i le x aq ui fo lia l . c au ca su s [1 3] w . h an sf or di i ( sy d. ) e . m ül l. sy n. a ca ro th al liu m h an sf or di i s yd . le av es o f r ub ia ce ae (c f. c an th iu m s p. ) u ga nd a [3 9] w . h ir tu lu s ( sp eg .) e. m ül l. sy n. d im er ie lla h ir tu la s pe g. ; a st er om yx a hi rt ul a (s pe g. ) th ei ss . & s yd . li ve le av es o f b ac ch ar is s pp . br az il, e cu ad or [1 3, 33 ,4 0– 42 ] w . i nc on sp ic uu s s po on er le af li tt er o f p oa fl ab el la ta so ut h g eo rg ia (a nt ar ct ic a) [4 3] w . i nd ic us t ila k, s .b . k al e & s .v .s . k al e le af li tt er o f i xo ra p ar vi flo ra in di a [4 4] w . j av an ic us k oo rd . sy n. d ic ha et is ja va ni ca (k oo rd .) c le m . & s he ar d ea d le av es o f f ic us e la st ic a (a ); de ad le av es of k en tia fo rs te ri an a (b ); le af s po t o n le af o f c yn od on d ac ty lo n (c ); le af s po t o n le af o f fi cu s e la st ic a (d ); le af s po t o n le af o f p er se a gr at is si m a (e ); so rg hu m v ul ga re (f ) (a ) i nd on es ia ( ja va ); (b ) f ra nc e (p ot p la nt ); (c –e ) b ur m a; (f ) p ap ua n ew g ui ne a (a ) [ 12 ,1 3] ; ( b ) [ 29 ]; (c –e ) [ 24 ]; (f ) [ 26 ] ta b. 1 su m m ar y of th e di st ri bu tio n an d su bs tr at um c ol on iz ed b y al l k no w n w en tio m yc es s pp . 242© the author(s) 2014 published by polish botanical society acta mycol 49(2):237–247 wilk et al. / wentiomyces sp. from plant litter sp ec ie s su bs tr at um lo ca lit y r ef er en ce s w . c fr . j av an ic us d ec om po si ng le av es o f p is ta ci a le nt is cu s, ph ill yr ea a ng us tif ol ia a nd c is tu s s pp . it al y [4 5] w . l ic he ni co la (h an sf .) d . h aw ks w . sy n. d im er ie lla li ch en ic ol a h an sf .; n eo co le ro a lic he ni co la (h an sf .) m .e . b ar r a st er ot hy ri um le uc op ht ha lm um o n le av es o f a ca ly ph a fr ut ic os a (a ) (a ) u ga nd a; (b ) u ni te d k in gd om (a ) [ 25 ,4 6] ; b [4 7] w . l ic he ni co la s ub sp . b ou te ill ei b ri ca ud , c l. r ou x & s ér us . sy n. n eo co le ro a lic he ni co la s ub sp . b ou te ill ei (b ri ca ud , c l. r ou x & s ér us .) m .e . b ar r fe llh an er a bo ut ei lle i o n le av es o f b ux us se m pe rv ir en s ( a ); ba ci di na s p. o n le av es o f bu xu s s em pe rv ir en s ( b ); ba ci di na c f. va sa ki i o n le av es o f b ux us se m pe rv ir en s ( c ); w oe ss ia s p. o n tr un k of s or bu s s p. (d ) (a –c ) s pa in ; ( a ) f ra nc e; (d ) l ux em bo ur g; (e ) u ni te d k in gd om ; ( f) u sa ; ( g ) a us tr ia (a –c ) [ 46 ]; (d ) [ 48 ]; (e –g ) [ 47 ] w . c f. lic he ni co la s ub sp . b ou te ill ei le ca ni a cu pr ea li th ua ni a [4 9] w . m el io lo id es (b er k. & m .a . c ur tis ) e . m ül l. sy n. a st er in a m el io lo id es b er k. & m .a . c ur tis ; d im er os po ri um m el io lo id es (b er k. & m .a . c ur tis ) g . m ar tin ; p ar od io ps is m el io lo id es (g . w in te r) m au bl .; c ha et os tig m e m el io lo id es (b er k. & m .a . c ur tis ) s yd . ba cc ha ri s h al im ifo lia (a ); le av es o f l ag en op ho ra pu m ila (b ); l. h ue ge lii (c ); l. st ip ita ta (d ); ba cc ha ri s s p. (e ); c on yz a sp . ( f) ; e ri ge ro n sp . (g ) (a ) u sa ; ( b ) n ew z ea la nd ; ( c ,d ) a us tr al ia ; (e –g ) n or th a nd s ou th a m er ic a, b er m ud a; (h ) b ra zi l (a ) [ 50 ]; (b ) [ 51 ]; (c ,d ) [ 26 ]; (e –g ) [3 3] ; h [4 7] w . m ol ar ife r sc he ue r sy n. n eo co le ro a m ol ar ife ra (s ch eu er ) m .e . b ar r d ea d le av es a nd s te m s of c ar ex d av al lia na (a ); c ar ex b al de ns is (b ) (a ) a us tr ia ; ( b ) g er m an y [5 2] w . o re op hi lu s ( sp eg .) e. m ül l. sy n. d im er os po ri um o re op hi lu m s pe g. ; v en tu ri a or eo ph ila (s pe g. ) th ei ss .; d im er ie lla o re op hi la (s pe g. ) e . m ül l. & a rx d ea d le av es o f r ho do de nd ro n fe rr ug in eu m a us tr ia , g er m an y [5 3, 54 ] w . p an de i s .k . b os e le av es o f c ra ta eg us c re nu la ta in di a [2 1] w . p el tig er ic ol a d . h aw ks w . sy n. r ac ib or sk io m yc es p el tig er ic ol a (d . h aw ks w .) m .e . b ar r; n ie ss lia p el tig er ic ol a (d . h aw ks w .) et ay o pe lti ge ra a ph th os a (a ); on d ea d po rt io ns o f th al li of p . l eu co ph le bi a (b ); p. r uf es ce ns (c ); p. br ita nn ic a (d ); pe lti ge ra s p. (e ); p. p ra et ex ta ta (f ); st er eo ca ul on d ep re ss um (g ); s. r iv ul ar is (h ); pe lti ge ra sc ab ro sa (i ); p. d id ac ty la ( j) (a ,c ,d ) n or w ay ; ( a ,d ,e ) g re en la nd ; ( a ) t he fa ro es ; ( a ,c ,h –j ) r us si a; (a ) a us tr ia ; ( a ,d ,g ) c an ad a; (a ,d ,f ) s pa in ; ( a ) s w ed en ; ( b ) g re at br ita in ; ( d ) u sa ; ( a ) i ce la nd (a ) [ 55 –7 1] ; ( b ) [7 2] ; ( c ) [ 57 ,6 3] ; ( d ) [6 5, 66 ,7 3– 75 ]; (e ) [7 3] ; ( f) [6 6] ; ( g ) [6 4] ; ( h ) [ 63 ,6 4] ; (i ,j) [6 3] ta b. 1 (c on tin ue d) 243© the author(s) 2014 published by polish botanical society acta mycol 49(2):237–247 wilk et al. / wentiomyces sp. from plant litter sp ec ie s su bs tr at um lo ca lit y r ef er en ce s w . s ib ir ic us (p et r.) e . m ül l. sy n. n eo co le ro a si bi ri ca p et r. d ry s te m s of v ac ci ni um m yr til lu s ( a ); de ad st em s of f ili pe nd ul a sp . ( b ); le av es o f l ar ix de ci du a (c ); va cc in iu m s p. (d ) (a ) r us si a (s ib er ia ); (b –d ) g re at b ri ta in (a ) [ 13 ,7 6] ; ( b, c ) [1 4] ; ( d ) [ 26 ] w . t at ja na e s. y. k on dr . sy n. n ie ss lia ta tja na e (s .y . k on dr .) et ay o pa ra sy m bi ot ic o n ps eu do cy ph el la ri a co ro na ta (a ); st ic ta c f. bo sc hi an a (b ); lo ba ri a sp . ( c ) (a ) t as m an ia , n ew z ea la nd ; ( b ) p ap ua n ew g ui ne a; (c ) n ot in di ca te d [7 7] w . v an en si s t ila k & t al de d ea d le av es o f f ic us b en gh al en si s in di a [2 1] w en tio m yc es s p. pa rm el ia c ap er at a (a ); de ad le av es o f r ho do de nd ro n to m en to su m (b ); c ro to n an tis yp hi lit ic us (c ); d ia ne lla re vo lu ta (d ); h ev ea b ra si lie ns is (e ); h ib is cu s e la tu s ( f) ; la ge no ph or a hu eg el ii (g ); pa nd an us s p. (h ); de ad le af o f e uc al yp tu s co cc ife ra (i ); de ad s te m o f c ha m er io n an gu st ifo liu m ( j) (a ) s pa in ; ( b ) p ol an d; (c ) b ra zi l; (d ,g ) a us tr al ia ; ( e) b ru ne i; (f ) v en ez ue la ; ( h ) m au ri tiu s; (i ,j) g re at b ri ta in (a ) [ 46 ]; (b ) [ 27 ]; (c –g ) [ 26 ]; (h ) [ 78 ]; (i ,j) [7 9] ta b. 1 (c on tin ue d) 244© the author(s) 2014 published by polish botanical society acta mycol 49(2):237–247 wilk et al. / wentiomyces sp. from plant litter communication, 2014). distribution of wentiomyces spp., together with substrata on which they were recorded, as compiled from the literature, is given in the tab. 1. strikingly, some species or specimens [e.g. w. alpivagus nograsek and w. fimbriatus (dearn. & house) m.e. barr, or w. fimbriatus and w. sibiricus (petr.) e. müller] were recorded from identical plant host species, raising a question about their true taxonomic status. there is an interesting suggestion about possible anamorph genus drawn by de hoog et al. [28], who have examined a herbarium material of w. javanicus deposited by joly [29]. joly [29] depicted anamorphic fungus, which he had isolated in pure culture from ascospores of his specimen of w. javanicus; de hoog et al. [28] noted the resemblance of this anamorph to rhinocladiella anceps (sacc. & ellis) s. hughes (recalled also by arzanlou et al. [30]). unequivocal link between these two morphs was however not demonstrated; moreover, according to molecular analyses species from the genus rhinocladiella and similar ramichloridium are placed predominantly within chaetothyriales and capnodiales, with possible teleomorphs for rhinocladiella in capronia spp. [30,31]. our specimens could not be identified to the species level, because we have not observed asci and ascospores, and the sole gross morphology of pseudothecia and setae, or host species, do not provide firm characters. wentiomyces oreophilus (speg.) e. müll. described from rhododendron ferrugineum l. has straight setae, and therefore cannot be considered conspecific with our finding (as was incorrectly assumed by wilk et al. [27] regarding another specimen from poor fen). wentiomyces sibiricus (petr.) e. müll., described from vaccinium myrtillus l. could be one possibility. worth noting is the fact, that the specimens presented here differ greatly from the specimen presented by wilk et al. [27], being in all respects smaller (42–50 μm versus 87.5–170 μm) and having less ramified setae. we cannot even state for sure, that both specimens reported here represent the same or different species, and therefore further research aimed specifically at habitatand hostpreferences of non-lichenicolous wentiomyces spp. is certainly needed. acknowledgments this work was partially financed by mnisw grant no. n305 052 240. funding from eu through the european social fund, contract number uda-pokl.04.01.01-00-072/09-00 to first author is also acknowledged. first author would like to express sincere thanks to b. pawlik (cracow), a. chlebicki (cracow), ch. scheuer (graz), n.v. filippova (khanty mansiysk), a. pierotti (ascofrance), b. wergen (ascofrance), m. bemmann (ascofrance), d.p. di lonardo (wageningen), m.m. thaung (los angeles) for invaluable help with obtaining important publications. two anonymous reviewers are acknowledged for invaluable comments greatly improving quality of the manuscript. authors’ contributions the following declarations about authors’ contributions to the research have been made: collected material, performed final identification, conducted literature study, prepared drawings and wrote the manuscript: mw; helped writing the manuscript: jp; designed the methods, performed initial identification, and helped writing the manuscript: mwr. references 1. thormann mn, rice av. fungi from peatlands. fungal divers. 2007;24:241–299. 2. gorham e, lehman c, dyke a, clymo d, janssens j. long-term carbon sequestration in north american peatlands. quat sci rev. 2012;58:77–82. http://dx.doi.org/10.1016/j.quascirev.2012.09.018 3. peltoniemi k, fritze h, laiho r. response of fungal and actinobacterial communities to water-level http://dx.doi.org/10.1016/j.quascirev.2012.09.018 245© the author(s) 2014 published by polish botanical society acta mycol 49(2):237–247 wilk et al. / wentiomyces sp. from plant litter drawdown in boreal peatland sites. soil biol biochem. 2009;41(9):1902–1914. http://dx.doi.org/10.1016/j. soilbio.2009.06.018 4. thormann mn. diversity and function of fungi in peatlands: a carbon cycling perspective. can j soil sci. 2006;86:281–293. http://dx.doi.org/10.4141/s05-082 5. thormann mn. the role of fungi in decomposition dynamics in peatlands. in: wieder rk, vitt dh, editors. boreal peatland ecosystems. berlin: springer; 2006. p. 101–123. 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available from: http://www.gbif.orgwww.fieldmycology.net/frdbi/frdbi.asp http://dx.doi.org/10.2307/3244200 http://dx.doi.org/10.2307/3244200 http://dx.doi.org/10.1127/0029-5035/2008/0086-0275 http://dx.doi.org/10.1016/s0007-1536(80)80167-7 http://www.gbif.orgwww.fieldmycology.net/frdbi/frdbi.asp abstract introduction material and methods descriptions of specimens discussion acknowledgments authors’ contributions references 2014-12-31t17:35:00+0000 polish botanical society 2014-08-28t14:43:36+0200 polish botanical society 2014-08-27t17:37:55+0200 polish botanical society 2014-08-25t14:57:06+0200 polish botanical society co-existence and interaction between myxomycetes and other organisms in shared niches irina o. dudka and katerina o. romanenko department of mycology, m.g. kholodny institute of botany national academy of sciences of ukraine tereshchenkivska 2, msp 1 kyiv, ua 01 001 dudka i.o., romanenko k.o. co existence and interaction between myxomycetes and other organisms in shared niches. acta mycol. 41(1): 99 112, 2006. the ecology of myxomycetes co existing with the latridiidae (coleoptera), bryophytes and ascomycetous, basidiomycetous and anamorphic fungi were studied in crimea and at different locations on the left bank of ukraine. results from the left bank indicate that the latridiidae feed on myxomycetes. colonies of the most common 13 myxomycete species (which included stemonitis axifera (bull.) macbr., s. fusca roth, s. splendens rost., fuligo septica (l.) wigg. and mucilago crustacea wigg.) were inhabited by 5 species of the latridiidae. myxomycete spores were present in guts of 19 of the 25 beetle specimens investigated. beetles latridius hirtus, enicmus rugosus and e fungicola seem to be obligate myxomycete feeders, while corticarina truncatella was clearly facultative. 13 species of myxomycetes were recorded on 9 species of moss and 3 species of liverwort developing on decaying wood or bark in the crimean nature reserve. relations between myxomycetes and bryophytes on woody substrata are spatial rather than trophic, and are possibly regulated by specific microclimatic conditions inside bryophyte thallomes. 69 species of myxomycetes were found co existing with 36 species of ascomycetes, 21 species of basidiomycetes and 9 species of anamorphic fungi in the crimean nature reserve. associations formed by myxomycetes and fungi on different woody substrata are analyzed. key words: myxomycetes, ascomycetes, basidiomycetes, anamorphic fungi, bryophytes, latridiidae beetles introduction in terrestrial ecosystems, certain patterns can be found governing the spatial distribution of living organisms. these are connected with trophic, topic and another relations. autotrophic plants are usually considered the foundation for the resulting consortia (ecological communities originating through co-development of heterogenous groups tightly joined by common activities), since they provide a prime food source, and a substratum on which other organisms can grow. myxomycetes (also known as slime moulds) often form a part of those consortia, and may also make associations with various other groups of organisms (d u d k a et al. 1976). research on acta mycologica vol. 41 (1): 99-112 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 100 i. o. dudka and k. o. romanenko the co-existence of slime moulds with beetles (coleoptera), bryophytes, ascomycetous, basidiomycetous and anamorphic fungi is briefly reviewed below. brief historical review myxomycetes are frequently associatiated with insects, especially beetles (coleoptera). s t e p h e n s o n et al. (1994) reviewed slime mould / beetle associations recorded since the end of the nineteenth century, and listed the beetle families most frequently forming these associations as follows: rhizodidae, leiodidae, staphylinidae, clambidae, eucinetidae, sphindidae, cerylonidae, latridiidae. the numerous and repeated records of these beetles on the same species of myxomycetes suggested an obligate connexion. since examination of beetle gut contents usually reveals abundant myxomycete spores, it is clear the beetles feed on them. in fact, some beetle species use not only spores but also plasmodia of slime moulds as a source of nutrition (l a w r e n c e , n e w t o n 1980; w h e e l e r 1980). the term myxomycetophagy has been proposed to describe the phenomenon of beetles deriving nutrition from slime moulds (n e w t o n 1984; n e w t o n , s t e p h e n s o n 1990). despite their rather diverse taxonomic range, slime moulds used by beetles as food share a number of morphological, developmental and ecological characters as follows: 1) abundant spores in large simple sporophores or large aethalia which comprise a group of sporangia covered with a common cortex and pillow-like; 2) a long period of sporophore formation during the growing season; 3) long-lasting sporophores; 4) abundance; 5) development on woody substrata; 6) spores with spiny, warty or net-like walls, 4 (5-9)-15 µm diam. (b l a c k w e l l 1984). arcyria incarnata (pers.) pers., stemonitis axifera (bull.) macbr., s. fusca roth, s. splendens rost., lycogala epidendrum (l.) fr., stemonaria longa (peck) nann.-brem., fuligo septica (l.) wigg., tubifera ferruginosa (batsch) gmel. and others largely match those criteria. s t e p h e n s o n et al. (1994) have shown that myxomycetes of the genus stemonitis roth are used as food by 10 beetle genera, fuligo hall. by 9, and arcyria wigg. and tubifera gmel. by 7 each. the feeding preferences of some beetles for myxomycetes explains their constant association with slime moulds. there is, however, evidence that these beetles also play some part in disseminating myxomycete spores. observation of fuligo septica spores from sphindidae beetle guts has shown that some spores pass through insect intestines without damage and can subsequently germinate, while transfer of spores of physarum straminipes lister and f. septica from the exoskeleton surface of latridiidae beetles to agar medium has also been observed, particularly when there are large numbers of such spores (b l a c k w e l l , l a m a n , g i l b e r t s o n 1 9 8 2 ). representatives of the family latridiidae (coleoptera) are small beetles, no more than 3 mm long, occurring under bark of fallen trunks, in decaying wood and in forest litter. worldwide about 750 species are known, 55 in ukraine. typically they are associated with fungi, in particular anamorphic fungi, ascomycetes and basidiomycetes, predominantly with basidiomata of wood-destroying fungi, where they feed on the spores (h o w a r d , c u r r i e 1932; n e w t o n , s t e p h e n s o n 1990; s t e p h e n s o n et al. 1994). information about associations between the latridiidae and myxomycetes has been summarized (d u d k a , tr i k h l e b , r o m a n e n k o 2002). the co-existence of myxomycetes and bryophytes has been studied much less than that of slide moulds and beetles. myxomycete sporophores at the thallome co existence and interaction between myxomycetes 101 surface of mosses and liverworts are rather widespread in nature (s t e p h e n s o n , s t e m p e n 1994; h ä r k ö n e n et al. 2002; s t o j a n o w s k a , p a n e k 2004). in most cases, however, associations of slime moulds and bryophytes arise accidentally since both components tend to occur on the same substratum – decaying and decomposing wood of deciduous and/or coniferous trees (s t e p h e n s o n , s t u d l a r 1985). it has also been surmised that myxomycetes develop more intensively and occur more frequently on fallen decaying logs overgrown with bryophytes because of their high humidity (s t o j a n o w s k a , p a n e k 2004). only a few myxomycetes, in particular barbeyella minutissima meylan, colloderma oculatum (lipp.) g. lister and lepidoderma tigrinum (schr.) rost. seem to be true bryophilous species. the first of these species is adapted to rich communities of liverworts, especially novellia curvifolia and species of the genus cephalozia which sometimes completely cover the surface of fallen decaying coniferous logs by (s c h n i t t l e r , n o v o z h i l o v 1998; s c h n i t t l e r , s t e p h e n s o n , n o v o z h i l o v 2000; n o v o z h i l o v 2005). in forest ecosystems many organisms are involved in decomposition of woody substrata. apart from slime moulds, wood-inhabiting fungi are also active at various stages of succession. some fungal species are known to develop and sporulate on the surface of myxomycete fruitbodies, using them as a feeding substratum (r o g e r s o n , s t e p h e n s o n 1993), but that is a case of fungal parasitism on slime moulds and an example of trophic relations which are probably negative for the myxomycete. other interrelations between myxomycetes and these fungi so far remain almost totally unstudied, although side-by-side co-existence of myxomycetes and fungi on the same woody substatum is rather commonly observed in nature. such relations appear to be indifferent, but that is only an assumption: exact information is absent not only about the nature of those relations but about also the slime mould and fungal species compositions on which they are based. co-existence of myxomycetes and true fungi has therefore been studied least of all (r o g e r s o n , s t e p h e n s o n 1993). objectives it is clear that the characteristics of the co-existence and of interrelations between myxomycetes and other organisms is still poorly understood. the aim of the present work has therefore been to contribute to that research by identifying myxomycetes associated with latridiidae family beetles, liverworts, mosses and wood-inhabiting anamorphic, ascomycetous and basidiomycetous fungi collected in different parts of southern ukraine. this study has also been designed to begin analysis of relations between component members in the associations. material and methods fieldwork was carried out from april to september 2000 on the left bank of ukraine within three geobotanical regions (donetsk, leftbank and starobilsk grassmeadow steppes), and in the crimean peninsula within the crimean nature reserve (montane crimea geobotanical region) during the growing seasons of 2000 and 2001. left bank myxomycete collections were predominantly from forest plantations near donetsk, and flood-plane forests around stanychno-luganske settlement in the valley of the siverski donets river (table 1) where oak (quercus robur l.), ash (fraxinus 102 i. o. dudka and k. o. romanenko excelsior l.) and aspen (populus tremula l.) are dominant tree species. beech (fagus sylvatica l.) and crimean pine (pinus pallasiana d. don) dominate other tree species in the study areas of forest communities in the crimean nature reserve. myxomycete colonies (groups of sporophores and aethalia) were sampled in the field from fallen trunks and logs of trees in various stages of wood decay, from fallen branches of various diameters and from decaying stumps. 53 field samples of myxomycetes were collected on woody substrata the left bank study areas and 450 from the crimean nature reserve. 300 samples of woody substrata from that reserve, mainly tree bark, were collected for laboratory detection of myxomycetes, using the moist chamber method (g i l b e r t , m a r t i n 1933; s t e p h e n s o n 1985; n a n n e n g a b r e m e k a m p 1991). for myxomycete identification, the main manuals on the group were used (n a n n e n g a b r e m e k a m p 1991; n o v o z h i l o v 1993; s t e p h e n s o n , s t e m p e n 1994). slime mould nomenclature follows i n g (1999). of the 53 samples collected from left bank forest plantations, 25 were examined for the presence of latridiidae in the myxomycete colonies. the biggest colonies were selected from the field samples, then divided into three more or less equal parts and placed in 90 mm sterile petri dishes with filter paper at the bottom. the dark bodies of beetles emerging from the colony were easily observed against the white surface of the paper. these individuals were then identified, and their gut contents sampled. for identification of beetles the appropriate volume of fauna hungariae was used (r ü c k e r 1983). preparations of gut contents for identification of myxomycetous spores were made in fore liquid (50 ml distilled water, 20 g chloralhydrate, 40 ml glycerine, 30 g gum-arabic). in addition to gut content studies of latridiidae specimens found on slime moulds collected in the field, living latridius hirtus were kept in the laboratory on aethalia of fuligo intermedia, to see if l. hirtus can complete its life cycle with only a single feed source present. pillow-like aethalia of f. intermedia, 0·5-4 cm long, up to 10 mm thick and undamaged by insects were collected in the field. these aethalia were checked for other insect inhabitants using a stereo microscope, then placed in sterile petri dishes and stored for two weeks to ensured absence of insects. adult latridius hirtus individuals were then added, and the dishes were regularly examined to follow development of the successive beetle stages. associations between myxomycetes and bryophytes were studied in the crimean nature reserve. special attention was paid to associations where slime moulds developed sporophores, aethalia or plasmodia directly on the surface of bryophyte thallus or inside it. associations between myxomycetes and wood-inhabiting fungi were studied in the crimean nature reserve to identify the component species and assess the influence of substratum size on species diversity. results interactions between myxomycetes and latridiidae beetles on woody substrata associations between myxomycetes and the latridiidae are shown in table 1. these associations were found to be rather widespread on the left bank of ukraine, co existence and interaction between myxomycetes 103 т а b l e 1 myxomycetes associated with latridiidae (coleoptera) at the left bank of ukraine myxomycete species, by order and family beetle species place and date of association collection beetle gut content liceales, reticulariaceae tubifera ferruginosa (batsch) gmel. reticularia lycoperdon bull. enicmus rugosus enicmus fungicola e. rugosus melanoph thalma sp. lugansk oblast, s. stan ychno luganske, 1 july 2000 donetsk area, 6 june 2000 donetsk area, 26 may 2000 lugansk oblast, s. stany chno luganske, 29 june 2000 spores of different myxomycete species including t. ferruginosa spores of r. lycoperdon spores of r. lycoperdon spores of alternaria alternata (hyphomycetes) trichiales, trichiaceae arcyria obvelata (oeder) onsberg e. rugosus lugansk oblast, s. stan ychno luganske, 30 june 2000 spores of a. obvelata stemonitales, stemonitaceae comatricha nigra (pers.) schroet. enerthenema papillatum (pers.) rost. stemonaria longa (peck) nann. brem. stemonitis axifera (bull.) macbr. stemonitis fusca roth stemonitis splendens rost. symphytocarpus amaurochaeoides nann. brem. e. rugosus e. rugosus e. rugosus e. rugosus corticarina truncatella e. rugosus c. truncatella e. rugosus latridius hirtus e. rugosus donetsk area, 18 august 2000 donetsk area, 3 september 2000 donetsk oblast, volodarski district, nature reserve kamiani mogyly, 22 july 2000 donetsk oblast, novo azovski district, azovske forestry, 3 july 2000 lugansk oblast, s. stan ychno luganske, flood plain forest, 2 july 2000 lugansk oblast, s. stan ychno luganske, flood plain forest, 2 july 2000 donetsk, 3d city pond, 8 august 2000 donetsk, 3d city pond, 8 august 2000 donetsk, 3d city pond, 8 august 2000 donetsk, putylivski forest, 18 july 2000 spores of c. nigra spores of e. papillatum spores of s. longa spores of s. axifera spores of alternaria alternata (hyphomycetes) spores of s. fusca spores of s. splendens spores of s. splendens spores of s. splendens spores of s. amaurochaetoides 104 i. o. dudka and k. o. romanenko with 5 species from 4 latridiidae genera being were found in the colonies of 13 species of myxomycetes from 10 genera. gut content analysis (from 5 of the latridiidae species encountered in colonies of the 25 investigated samples of 13 myxomycete species) revealed slime moulds spores in the guts of 19 individuals (tab. 1). in 16 of those cases, the spores found belonged to the slime mould species on which the beetle was collected. spores of tubifera ferruginosa, fuligo septica and mucilago crustacea were found in the guts of enicmus rugosus specimens not originating from colonies of those species. other spores distinct in morphology and size were also present in the guts of those enicmus rugosus specimens. these may also have been myxomycete spores, but their taxonomic position was not established. no slime mould spores were found in the guts of female melanophthalma sp. individuals found on reticularia lycoperdon or from corticarina truncatella individuals collected on stemonitis fusca; instead, the guts of those beetles were filled with spores of the anamorphic fungus alternation alternata (fr.) keisler. study of the living latridius hirtus, kept in the laboratory on aethalia of fuligo in termedia, resulted in the following sequence of observations: adults, laying eggs in the aethalia, larvae, moult, pupae, young adults emerging from pupae. co-existence of myxomycetes and bryophytes on woody substrata altogether 13 species of myxomycetes were recorded on 9 species of moss and 3 species of liverwort associated with woody substrata in the crimean nature reserve (table 2). arcyria cinerea (bull.) pers., echinostelium arboreum keller et brooks, e. minutum de bary, macbrideola cornea (g. lister et cran) alexop., perichaena vermicularis (schw.) rost., physarum cinereum (batsch) pers. were most commonly encountered in association with the following bryophytes widespread in montane crimea: hypnum cupressiforme, leucodon sciuroides and porella platyphylla. didymium physarales, physaraceae fuligo intermedia macbr. fuligo septica (l.) wigg. e. rugosus l. hirtus e. rugosus l. hirtus donetsk area, 3 september 2000 donetsk area, 3 september 2000 donetsk, putylivski forest, 18 july 2000 donetsk, valley of the river bakhmutka, 27 june 2000 spores of f. intermedia spores of f. intermedia spores of different myxomycete species including f. septica spores of f. septica physarales, didymiaceae mucilago crustacea wigg. l. rugosus l. hirtus donetsk area, 10 july 2000 donetsk area, 3 september 2000 spores of different myxomycete species including m. crustacea spores of m. crustacea tab. 1 cont. co existence and interaction between myxomycetes 105 trachysporum g. lister on ctenidium molluscum, licea minima fr. on hypnum cupressiforme, perichaena chrysosperma (currey) lister on frullania dilata, stemonitis fusca on leucodon sciuroides, symphytocarpus amaurochaetoides on pterigynandrum filiforme, s. impexus ing. et nann.-brem. on porella platyphylla and trichia varia (pers.) pers. on anomodon viticulosus were more rare in bryophytes communities of the reserve (tab. 2). augmenting information about woody substrata in table 2, echinostelium minutum was collected on bark of acer stevenii pojark., alnus glutinosa (l.) gaertn., carpinus betulus l., fagus sylvatica, pinus kochiana klotzsch., p. pallasiana, p. sylvestris l., quercus petraea (mattuschka) liebl., q. robur, sorbus torminalis (l.) crantz., macbrideola cornea was collected on bark of acer campestre l., acer stevenii, carpinus betulus, cornus mas l., crataegus sp., fagus sylvatica, fraxinus excelsior, taxus baccata l., tilia cordata mill. and perichaena vermicularis was collected on bark of acer campestre, alnus glutinosa, cornus mas, fraxinus excelsior, pyrus communis l., q. petraea, taxus baccata, tilia cordata. on the reserve didymium trachysporum was associated with ctenidium molluscum but there are additional records of it on fallen conifer needles, pieces of small bushes and herbivore dung. physarum cinereum was found on fallen leaves and decaying wood, but was most frequent in bryophyte communities as listed in table 2. ta b l e 2 myxomycetes collected in the crimean nature reserve on bryophytes species of myxomycetes species of bryophytes woody substrata arcyria cinerea (bull.) pers. frullania dilatata hypnum cupressiforme porella platyphylla absence didymium trachysporum g. lister ctenidium molluscum echinostelium arboreum keller et brooks leucodon sciuroides hypnum cupressiforme echinostelium minutum de bary hypnum cupressiforme leucodon sciuroides pterigynandrum filiforme racomitrium canescens absence licea minima fr. hypnum cupressiforme decaying wood macbrideola cornea (g. lister et cran) alexop. leucodon sciuroides porella platyphylla pterigynandrum filiforme hypnum cupressiforme metzgeria furcata absence perichaena chrysosperma (currey) lister frullania dilatata perichaena vermicularis (schw.) rost. hypnum vaucheri porella platyphylla absence physarum cinereum (batsch.) pers. anomodon viticulosus homolothecium sericeum hypnum cupressiforme porella platyphylla pseudoleskeella nervoza absence stemonitis fusca leucodon sciuroides decaying wood symphytocarpus amaurochaetoides pterigynandrum filiforme decaying wood symphytocarpus impexus ing et nann. brem. porella platyphylla trichia varia (pers.) pers. anomodon viticulosus decaying wood 106 i. o. dudka and k. o. romanenko co-existence of myxomycetes and fungi on woody substrata in the crimean nature reserve, 69 species of myxomycetes and 66 species of fungi from the ascomycota (36), basidiomycota (21) and anamorphic fungi (9) were collected on decaying woody substrata of coniferous and broadleaf trees. all fungi recorded were species usually regarded as wood-inhabiting. various complexes could be distinguished according to substratum type (tab. 3) and range of associated species (tab. 4). altogether 52 associations (tab. 3) were encountered in the crimean nature reserve between slime moulds and wood-inhabiting fungi, including 7 on large fallen trunks of trees, 34 on fallen branches of various diameters and 11 on decaying stumps. the highest numbers of myxomycete and the fungal species (from 13 to 35) occurred in associations on large fallen tree trunks. species diversity was significantly less (from 2 to 8) in associations on fallen branches (from 5 to 15 cm in diameter) and on decaying stumps. the following fungi were dominant (tab. 4) in associations on small woody remnants and on large fallen decaying tree trunks. pyrenomycetes: ceratostomella cirrhosa (pers.) sacc., lasiosphaeria hirsuta (fr.) ces. et de not., melanomma pulvis-pyrius (pers.) fuckel, nectria peziza (tode) fr., rosellinia conglobata (fr. et fuckel) sacc.; discomycetes: bisporella citrina (batsch) korf et s.e. carp., mollisia melaleuca (fr.) sacc., orbilia coccinella (sommerf.) fr. these were associated with the following myxomycetes: arcyria denudata (l.) wettst., ceratiomyxa fruticulosa (mull.) macbr., hemitrichia clavata (pers.) rost., lycogala epidendrum, metatrichia vesparium (batsch) nann-brem., physarum nutans pers., stemonitis fusca, stemonitopsis typhina (wigg.) nann-brem., trichia decipiens (pers.) macbr., t. scabra rost. and t. varia (pers.) pers. the basidiomycete calocera cornea (batsch) fr. and the anamorphic fungus brachysporium nigrum (link: fr.) s. hughes often joined those associations. fomes fomentarius (l.: fr.) fr., stereum hirsutum (willd.) fr., some species of the genus xylaria hill ex schrank [particularly x. longipes nitschke and x. polymorpha (pers.) grev.] were also found in those assocations, but only on large fallen trunks. all listed species of myxomycetes and fungi formed the main body of associations on the woody substrata in the reserve. frequency of occurrence in associations was high throughout the period of study. sometimes pyrenomycetes amphisphaeria umbrina (fr.) de not., strickeria kochii koerb., lasiosphaeria ovina (pers.) ces. et de not., discomycetes mollisia cineta b l e 3 distribution of associated complexes between myxomycetes and xylotrophic fungi on the different types of woody substrates type of woody substrate number of associated complexes between myxomycetes and wood inhabiting fungi range of species from both groups in associated complexes large fallen tree trunks 7 from 13 to 35 fallen branches of trees 34 from 2 tо 6 stumps 11 from 2 to 8 total 52 133 co existence and interaction between myxomycetes 107 rea (batsch) p. karst., trichophaeopsis bicuspis (boud.) korf et erb., basidiomycetes schizopora paradoxa (schrad.: fr.) donk, phellinus ferruginosus (schrad.: fr.) pat., fibuloporia mucida (pers.: fr.) niemelä, anamorphic fungi pleurothecium recurvatum (morgan) höhn., periconia cambrensis e.w. mason et m.b. ellis, heteroconium chaetospira (grove) m.b. ellis were also present on the numerous woody remnants ta b l e 4 some complexes of myxomycetes and wood inhabiting fungi on woody substrata differenti ated by size myxomycete species species of wood inhabiting fungi associated complexes on large fallen tree trunks arcyria denudata fuligo septica lycogala epidendrum stemonitis fusca stemonitis flavogenita jahn trichia scabra trichia varia ceratostomella cirrhosa, c. ampullasca (cooke) sacc., nectria coccinea (pers.) fr., n. peziza, lasiosphaeria hirsuta, l. ovina, melanomma pulvis pyrius, coniochaeta ligniaria (grev.) massee, rosellinia conglobata, xylaria longipes, strickeria kochii, dasyscyphus virgineus s.f. gray, bisporella citrina, mollisia melaleuca, orbilia coccinella, trichophaeopsis bicuspis, calocera cornea, fibuloporia mucida, stereum hirsutum, fomes fomentarius, marasmius alliaceus (jacq.) fr., oudemansiella sp., armillaria mellea (vahl.) p. kumm.,, pleurothecium recurvatum, pseudospiropes subuliferus (corda) m.b. ellis, acrogenospora sphaerocephala (berk. et broome) m.b. ellis, graphium calicioides (berk.) cooke et massee lycogala epidendrum metatrichia vesparium stemonitis fusca trichia varia lasiosphaeria hirsuta, nectria peziza, melomastia mastoidea (fr.) schroet., melanomma pulvis pyrius, chaetosphaeria ovoidea sacc., mollisia melaleuca, bisporella citrina, xylaria polymorpha, calocera cornea, bjerkandera adusta (willd. ex fr.) p. karst., stereum hirsutum, fomes fomentarius, pleurotus ostreatus (jacq.) p. kumm., pleurothecium recurvatum, periconia cambrensis arcyria denudata comatricha laxa rost. hemitrichia calyculata (speg.) farr hemitrichia clavata (pers.) rost. lycogala epidendrum stemonitis fusca symphytocarpus amaurochaetoides trichia scabra trichia varia melanomma pulvis pyrius rosellinia conglobata lasiosphaeria hirsuta bisporella citrina trichophaeopsis bicuspis fomes fomentarius associated complexes on fallen branches of trees trichia decipiens ceratostomella cirrhosa, mollisia melaleuca, orbilia coccinella, microthelia incrustans (el. et ev.) corlet et s. hughes as anamorph dendryphiopsis sp. arcyria cinerea (bull.) pers. nectria coccinea, n. peziza, eutypa lata (pers.) tul. et c. tul., orbilia coccinella, trichophaeopsis bicuspis hemitrichia clavata trichia decipiens rosellinia conglobata, chaetosphaeria ovoidea, mollisia melaleuca, calocera cornea associated complexes on the stumps comatricha nigra enerthenema papillatum hypocrea citrina (pers.) fr., mollisia lignicola phill., stereum hirsutum ceratiomyxa fruticulosa trichia decipiens lasiosphaeria hirsuta, amphisphaeria umbrina, mollisia cinerea lycogala epidendrum bisporella citrina, stereum hirsutum, brachysporium nigrum 108 i. o. dudka and k. o. romanenko of various sizes, but these fungi were rare in comparison with the dominant species: they were often represented by a single record in the reserve. discussion interactions between myxomycetes and latridiidae beetles on woody substrata outside ukraine, reticularia lycoperdon (liceales, reticulariaceae), various species of the genus stemonitis (stemonitales, stemonitaceae), in particular s. fusca, and more rarely fuligo septica (physarales, physaraceae) are the most prevalent slime moulds in associations with the latridiidae, while the genera enicmus and dienerella are the most wide-spread representatives of the family latridiidae to be involved in associations with slime moulds (n e w t o n , s t e p h e n s o n 1990; s t e p h e n s o n et al. 1994). within ukraine, the same myxomycete and latridiidae taxa were dominant in associations on the left bank. the myxomycete family stemonitaceae (stemonitales) was best represented, with 7 species from 5 genera, associated with 3 species of the latridiidae, enicmus rugosus being dominant (tab. 1). myxomycetes from the order physarales were present in left bank ukraine associations with the latridiidae as follows (3 species from 2 genera and 2 families): fuligio intermedia, f. septica (physaraceae) and mucilago crustacea (didymiaceae). they were associated with 2 species of the latridiidae – e. rugosus and latridius hirtus. both beetle species were associated with each of the 3 physarales species. reticularia lycoperdon (liceales) was found from 3 locations in association with enicmus fungicola, e. rugosus and melanophthalma sp. (distinguenda-group an exact species identification was not possible from the females collected). interestingly, r. lycoperdon has been widely reported as the basic myxomycete component in associations with the latridiidae in many parts of the world. among the myxomycetes connected with latridiidae beetles in our left bank sampling sites, large aethalia (0·5-20 cm long and 0·1-3 cm thick) are typical for reticularia lycoperdon, fuligo intermedia, f. septica and mucilago crustacea. sporophores closely grouped in large and dense colonies are observed in species of the genus stemonitis (especially s. fusca and s. splendens). sporophores of symphytocarpus amaurochaetoides tend to coalesce resulting in the formation of complex fruiting structures. the sporophores and aethalia of these slime moulds therefore provide sufficient spores to attract feeding by la tridiidae beetles. together with stemonaria longa, comatricha nigra and some other species, they all have a long period of sporophore formation and the sporophores so formed then tend to be long-lasting. furthermore, all of these slime mould species develop on woody substrata and have spores from 4-15 µm diam. with ornamented walls. they therefore have morphological, developmental and ecological characters which fit the criteria proposed by b l a c k w e l l (1984) for slime moulds associated with beetles. the presence of myxomycete spores in latridiidae gut contents shows that these beetles eat not only true fungi but also myxomycetes, while our observations of latridius hirtus kept alive in the laboratory in defined conditions showed that at least one species is able to complete its life cycle, apparently normally, with only aethalia of one myxomycete species, f. intermedia, as food. co existence and interaction between myxomycetes 109 there are very few reports from other parts of the world of myxomycete feeding by latridiidae beetles. a few individuals of an unknown species of enicmus were collected on colonies of reticularia lycoperdon from four locations in himachal pradesh (india): the gut contents of two were analysed and spores of r. lycoperdon found in both. a few individuals of a previously undescribed species of dienerella were discovered on colonies of stemonitis fusca from two locations in himachal pradesh: the guts content of an individual from each location were examined, and in both cases the guts were filled with spores of s. fusca (n e w t o n , s t e p h e n s o n 1990). our results on for latridiidae beetles and myxomycetes in ukraine are entirely compatible with those observations. our work therefore confirms that, in addition to the cerylonidae, leiodidae, scaphidiinae (staphylinidae), sphindidae and some other beetle families, the latridiidae also feed on myxomycetes. there is some question about whether the latridiidae are obligate or facultative feeders on myxomycetes. n e w t o n and s t e p h e n s o n (1990) considered their new species of dienerella from india to be a facultative feeder. results from the left bank of ukraine suggest that latridius hirtus, enicmus rugosus and e. fungicola may be obligate feeders: l. hirtus, in particular, has been shown to be capable of completing its life cycle with only myxomycetes as a food supply. corticarina truncatella, however, is more likely to be facultative: records on myxomycetes are rather rare, and it was usually found in forest litter, and on hay and other plant remains; furthermore gut content analysis shows this species can feed on fungal spores as well as those of slime moulds. use of myxomycetes by latridiidae beetles extends beyond feeding: they may lay their eggs in sporophores, aethalia and plasmodia; large downy colonies of myxomycetes may provide good cover for larvae and adults. the absence of myxomycete spores in guts of melanophthalma sp. distinguenda-group beetles from colonies of reticularia lycoperdon suggests that the association between these beetles and myxomycetes is based on something other than a food chain perhaps a spatial relationship although that needs further study. co-existence of myxomycetes with bryophytes on woody substrata decaying fallen trunks, logs, large branches, decomposing stumps are good substrata for simultaneous development of a diverse biota, including slime moulds, liverworts and mosses. myxomycetes often occur on wood overgrown with bryophytes (s t e p h e n s o n , s t u d l a r 1985; s t e p h e n s o n , s t e m p e n 1994; s c h n i t t l e r , n o v o z h i l o v 1998; s c h n i t t l e r , s t e p h e n s o n , n o v o z h i l o v 2000; h ä r k ö n e n et al. 2002). woody substrata covered with bryophytes provide favourable conditions for development of slime moulds, particularly on account of their high humidity (s t o j a n o w s k a , p a n e k 2004; n o v o z h i l o v 2005). all slime mould species found in the reserve associated with mosses and liverworts are usually considered lignicolous or corticolous: licea minima, stemonitis fusca, symphytocarpus amaurochaetoides and trichia varia, usually collected from decaying wood (i n g 1999), were also found on that substratum in the reserve. arcyria cinerea, echinostelium arboreum, e. minutum, macbrideola cornea, perichaena vermicularis and physarum cinereum are known as typical corticolous species (m i t c h e l l 1980). in our study area most of these corticolous species also were more often noted on 110 i. o. dudka and k. o. romanenko bark of various trees (tab. 2). echinostelium arboreum was the exception. in the reserve, every collection was made on moss, predominantly leucodon sciuroides but once hypnum cupressiforme. didymium trachysporum and physarum cinereum are well documented as species developing on plant remains in the litter (i n g 1999), but there is also a report of their occurrence in association with bryophytes (s t e p h e n s o n , s t u d l a r 1985). our results from crimea match this pattern exactly. some slime mould species (arcyria cinerea, echinostelium arboreum, e. minutum, macbrideola cornea, perichaena vermicularis and physarum cinereum) developed on more than one bryophyte species. this may be evidence that some associations between myxomycetes and bryophytes are purely accidental, arising because slime moulds and bryophytes are both well-adapted to develop on the same substrata (bark or wood). the associations may, however, alternatively arise because bryophytes provide ideal protection and other ecological conditions in which slime moulds can complete their full life cycle from plasmodia to sporophores. the relation between myxomycetes and bryophytes can be regarded as one of shared space. the bryophytes ensure temperature and humidity conditions suitable for slime mould development. fluctuations of temperature and humidity are less inside the bryophyte thallomes than outside, so that the bryophytes function like a moist chamber for the slime moulds, providing favourable conditions for their development. this function benefits not only the myxomycetes but also the bacteria, algae and protozoa on which they feed (s t e p h e n s o n , s t e m p e n 1994). the myxomycetes produce sporophores and aethalia from plasmodia in the drier areas on the bryophyte open surfaces. bryophytes associated with myxomycetes thus seem neither to benefit nor be damaged by them (s t e p h e n s o n , s t u d l a r 1985): bryophytes associated with slime moulds continue normal growth and reproduction, while the myxomycetes have a favourable biotope in the bryophytes communities. interrelations between the bryophytes and the slime moulds are thus generally positive and may be classified as commensal: unilateral use of one species by another without damage. co-existence of myxomycetes and fungi on woody substrata large fallen tree trunks might be thought to provide the richest associations of slime moulds and wood-inhabiting fungi, but in fact most associations (34) were recorded on fallen branches (tab. 3). direct contact of fallen branches with soil allows water to accumulate in the wood. shade and grass cover makes loss of humidity from fallen branches and temperature fluctuations in the lower layer of forest vegetation less than from large fallen trunks and stumps. the hydrothermal regime of the fallen branches and their location positively influenced formation of a significant number of associations between myxomycetes and wood-inhabiting fungi. such associations on woody substrata of various sizes and volumes are not directly connected and can exist independently. the biotic relations within those associations can therefore be characterized as neutral. species diversity in myxomycete / fungal associations on woody substrata can be compared according to size and volume of the substrata sampled (tab. 4). one possible explanation for the correlation between species diversity and size (diameter) co existence and interaction between myxomycetes 111 of the woody substratum may be that larger woody remnants conserve water longer thus providing both myxomycetes and wood inhabiting fungi with better conditions for development. substratum size may also determine the amount and availability of nutrients necessary for growth and formation of fruitbodies. conclusions co-existence and interaction between slime moulds, beetles, bryophytes and fungi have specific features for every pair of interacting organisms: at one end, the latridiidae may be either obligate or facultative feeders on slime moulds; at the other, relations between slime moulds and wood-inhabiting fungi are neutral. further studies are necessary to clarify the mechanisms regulating these biotic relations. acknowledgements: we would like to thank dr tetiana trikhleb from the national university in donetsk (ukraine) for identification of latridiidae, and dr svitlana nyporko from the department of lichenology and bryology, m.g. kholodny institute of botany nasu in kyiv (ukraine) who 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ridae. magyarország állatvilága (fauna hungariae) 158. budapest. 68 pp. s c h n i t t l e r m . , n o v o z h i l o v y u . k . 1998. late autumn myxomycetes of the northern ammer gauer alps. nova hedwigia. 66 (1 2): 205 222. s c h n i t t l e r m . , s t e p h e n s o n s . l . , n o v o z h i l o v y u . k . 2000. ecology and world distribu tion of barbeyella minutissima (myxomycetes). mycol. res. 104: 1518 1523. s t e p h e n s o n s . l . 1985. slime molds in the laboratory ii. moist chamber cultures. amer. biol. teach er 47: 487 489. s t e p h e n s o n s . l . , s t e m p e n h . 1994. myxomycetes: a handbook for slime molds. portland, oregon: timber press. 183 pp. s t e p h e n s o n s . l . , s t u d l a r s . m . 1985. myxomycetes fruiting upon bryophytes: coincidence or preference? j. bryol. 13: 537 548. s t e p h e n s o n s . l . , w h e e l e r q . d . , m c h u g h j . v. , f r a s s i n e t p. r . 1994. new north american associations of coleoptera with myxomycetes. j. nat. hist. 28: 921 936. s t o j a n o w s k a w. , p a n e k e . 2004. myxomycetes of the nature reserve near wałbrzych (sw po land) part ii. dependence on the substrate and seasonality. acta mycol. 39 (2): 147 159. w h e e l e r q . d . 1980. studies on neotropical slime mold/beetle relationships. part i. natural history and description of a new species of anisotoma from panama (coleoptera: leiodidae). proc. ento molog. soc. washington 82: 493 498. współistnienie i interakcje pomiędzy śluzowcami a innymi organizmami s t r e s z c z e n i e w artykule podane są wyniki badań mikoekologicznych na terenie krymu i ukrainie le wobrzeżnej. określono status taksonomiczny śluzowców, które współistnieją z owadami z ro dziny latridiidae (coleoptera), mchami, grzybami: ascomycota, basidiomycota i grzybami anamorficznymi. badania stosunków śluzowców i owadów z rodziny latridiidae w stepowych regionach ukrainy lewobrzeżnej ukazały istnienie troficznego związku owadów i śluzowców. w koloniach 13 gatunków śluzowców, wśród których przeważały: stemonitis axifera, s. fusca, s. splendens, fuligo septica i mucilago crustacea, występowało 5 gatunków owadów z rodziny latridiidae. analiza treści żołądkowej ukazała istnienie zarodników śluzowców w przewodach pokarmowych 19 z 25 przebadanych osobników. stwierdzono, że owady latridium hirtus, en cimus rugosus, e. fungicola są myxomycetofagami obligatoryjnymi, a corticarina truncatella może być uważany za myxomycetofaga fakultatywnego. 12 gatunków śluzowców podanych było z powierzchni 12 gatunków mchów, które rozwjiały się na butwiejącym drewnie lub na korze w krymskim rezerwacie przyrody. stosunki śluzowców i mchów, wspólnie rosnących w rezerwacie, na jednym drzewnym substracie są scharakteryzowane jako topiczne. są one regulowane przez specyficzne mikroklimatyczne warunki, które tworzą się w środku mchów z wysoką wylgotnością. stwierdzono wspólne występowanie 69 gatunków śluzowców z 36 ga tunkami workowców, 21 gatunkami podstawczaków, 9 gatunkami anamorficznych grzybów. analizowany skład asocjacji, które tworzą śluzowce z grzybami zależy od rozmiarów i objęto sci substratów drzewnych. 2014-01-01t11:43:46+0100 polish botanical society xylaria oxyacanthae tul. et c. tul., a new species for poland anna kujawa1 and dariusz karasiński2 1field station of research center for agricultural and forest environment polish academy of sciences turew, szkolna 4, pl-64-000 kościan, annakuja@poczta.onet.pl 2 staromostowa 4/8, pl-30-506 kraków, darek_karasinski@op.pl k u j a w a a., k a r a s i ń s k i d.: xylaria oxyacanthae tul. et c. tul., a new species for poland. acta mycol. 42 (1):75-78, 2007. the paper presents information about xylaria oxyacanthae tul. & c. tul., a new species for poland. the fungus was for the first time found in the mid-field shelterbelt near turew village and in the city park in kraków. key words: ascomycetes, xylariales, macrofungi, xylaria oxyacanthae, distribution, poland introduction xylaria oxyacanthae tul. et c. tul. belongs to xylariales (k i r k et al. 2001). for the first time it was described in 1863 from france. the species, grows on mummified fruits of crataegus monogyna therefore its distribution is closely related to occurrence of the species. in europe is rarely seen, among others in england (m i n t e r 1986), netherlands (b a s 1981; r e y n d e r s 1983), czech republic (a n t o n í n , va g n e r 1998), germany (k r i e g l s t e i n e r 1983, 1993; h e t t i c h , b e e n k e n 1997), denmark (l æ s s ø e 1994; w h a l l e y 2000), belgium (d e m e u l d e r 1984) and france (tu l a s n e , tu l a s n e 1863; l e r o y , s u r a u l t 1995). there are also some localities found in north america (s t o w e l l , r o g e r s 1983). unlike neighboring germany and czech republic (as mentioned above) in poland the species has never been recorded before. description of the species morphology like other species of the genus, xylaria oxyacanthae passes through two stages – anamorphic and teleomorphic one. the development of stromata begins in spring. the stromata develop on fallen and usually buried fruits of hawthorn that were produced in the year before. in summer, conidiophores with conidia develop on stromata (figs 1b, 1c; 2a). during the late summer and autumn the same stromata form acta mycologica vol. 42 (1): 75-78 2007 dedicated to professor alina skirgiełło on the occasion of her ninety-fifth birthday 76 a. kujawa and d. karasiński the basis for perithecia and those, when mature, produce the ascospores (figs 1a; 2c). the stromata die out during the late autumn. the macroand microscopic features of x. oxyacanthae have been described by many authors (tu l a s n e , tu l a s n e 1863; b a s 1981; s t o w e l l , r o g e r s 1983; h e t t i c h , b e e n k e n 1997; a n t o n í n , va g n e r 1998). macroscopic features. stromata which grow up from mummified fruits of hawthorn are slender, simple or branched, 10-60 mm long and 1-4 mm thick, cylindrical, lightly laterally compressed. in conidial stage conidiophores develop on upper part of stromata and produce conidia which cover stromata with white layer. apices yellowish to pink, remain sterile. lower parts of stromata remain black and sterile too (figs 1b, 1c; 2a). during anamorphic stage stromata grow thicker (3-4mm), and globose or subglobose perithecia (0.3-0.5 mm in diameter) submersed in stromata become visible. in this period stromata are easy to be overlooked as they become black and their brighter apices often come away (figs 1a; 2c). the lower parts of stromata are covered in the soil. the hawthorn fruits they grow upwards from are sometimes join together by very thin filaments of hyphae. microscopic features. conidia globose to ovoid, uncoloured, 3.1-4.3 x 2.4-3.8 μm. asci 8-spored, cylindrical. according to b a s (1981) size of asci amounts 78-94 x 4.58.5 μm, but according to other authors (s t o w e l l , r o g e r s 1983; h e t t i c h , b e e n k e n 1997; a n t o n í n , va g n e r 1998) they could be somewhat bigger: 100-140 x 6-9 μm. apical apparatus cylindrical, iodine positive. paraphyses filamentous, lucid. ascospores inaequilaterally elliptic to navicular, dark brown to black, with straight germ slit running almost trough the full length of the spore (8) 10-13 (14) x 4.5-6.0 μm. macroand microscopic features of x. oxyacanthae found in poland do not differ from those described in bibliography. the size of asci amounts to 120-135 x 6-8 μm and fit the range given by most of other authors. characteristic of the localities for the first time x. oxyacanthae was found in poland on 5th of july 2001 during the long term research on macromycetes of the general dezydery chłapowski landscape park (fig. 1). about 300 of anamorphic stromata growing on mummified hawthorn fruits were discovered in a midfield shelterbelt near turew (atpol bd 48-00).the plant community composition of this afforestation is similar to riparian forest populetum albae. the canopy is dominated by populus alba, the shrub layer by crataegus mono­ gyna with some admixture of cornus sanguinea and sambucus nigra. the herb layer is formed by mesophilous species from querco­fagetea class, such as aegopodium podagraria, brachypodium sylvaticum, ficaria verna, festuca gigantea, ranunculus lanuginosus, stachys sylvatica. the representatives of nitrophilous species from artemisietea vulgaris class are also present: alliaria petiolata, galium aparine, geum urbanum and urtica dioica. during the few consecutive years of research following numbers of x. oxyacan­ thae stromata were recorded on the site: in 2002 – 300, in 2003 – 5, in 2004 – 200, and in 2005 – 600. the hawthorn is a quite common species in the landscape park, xylaria oxyacanthae 77 but in spite of very careful search no other localities of the fungus were found in the area. the second polish locality of xylaria oxyacanthae was discovered on 21th of june 2004 in a city park called „skały twardowskiego” in krakow (atpol df-69). the anamorphic stromata of the fungus were found at the park edge close to tyniecka street, some 30 m away from the bank of vistula river (fig. 2). the site is impure by strollers and presumably no intensive green-keeping works have recently been carried out there, a part from periodical shrubs trimming along the footpath. the tree stand is diverse with predomination of fraxinus excelsior and acer platanoides, and some admixture of acer negundo, betula pubescens, malus domestica, juglans regia, prunus avium, robinia pseudacacia and tilia cordata. shrub layer is well developed, with predomination of crataegus monogyna, sambucus nigra and undergrowth of acer platanoides, a. negundo and fraxinus excelsior, additionally enriched by phila­ delphus coronarius. despite of deep shade, herb layer is very abundant covering 80% of the plot. nitrophilous species from the artemisietea vulgaris class such as cheli­ donium majus, urtica dioica, geum urbanum, galium aparine, alliaria petiolata and viola odorata are most frequent. woodland species from the querco­fagetea class have also significant share, especially ficaria verna and aegopodium podagraria. stromata of xylaria oxyacanthae grew on the verge of shallow depression under one of the several hawthorns. altogether 50 differently developed specimens (the biggest one was 27 mm tall) were found on the area of 2 m2. most of the stromata were rachitic and gracile, and only in few cases branched. the fruits of crataegus mo­ nogyna colonized by xylaria oxyacanthae were most often buried several cm under the ground and linked to stromata by thin, black filaments of hyphae. in several cases the filaments growing upwards from two or three mummified fruits were joined together to form single stromatum. only in few cases the stromata grew upwards from the fruits that were not fully buried in the substrate. one-half of the stromata was collected and the rest was left, but despite of intensive search no teleomorphic forms of stromata were found in autumn. in the spring 2005, on 17th of may, a dozen or so of new stromata were found exactly in the same place. during the next few visits it was observed that the stromata have gradually disappeared, as the result of intensive feeding of arthropods from the armadilidium vulgare species (fig. 2b), which occurred in masses. remarks some authors suggest that the species is not as rare as it could be inferred from only few localities found, because, as they underline, due to small size, dark colour of stromata and specific environmental requirements the species is easy to be overlooked (m i n t e r 1986, l æ s s ø e 1994). other authors (w h a l l e y 1987; l e r o y , s u r a u l t 1995) stress, that first of all, the specific habitat demands are the reason for rare occurrence of the species, as its distribution is absolutely dependent on buried hawthorn fruits presence. studies carried out in france (l e r o y , s u r a u l t 1995) and germany (k r i e g l s t e i n e r 1993; h e t t i c h , b e e n k e n 1997) showed that the occurrence of hawthorn per se is insufficient for x. oxyacantha occurence. additional conditions favorable for x. oxyacantha presence are either the thick layer of litter deposition, that can cover hawthorn fruits, or the presence of soil 78 a. kujawa and d. karasiński cracks into which the fruits can fall, because the fungus stromata most often grow from the fruits buried shallow under the ground surface (b a s 1981; r e y n d e r s 1983; l e r o y , s u r a u l t 1995). the other interesting feature characteristic for the species ecology are the numerous stromata which sometimes occur in as many as hundred per one m2 (r e y n d e r s 1983; d e m e u l d e r 1984; h e i t t i c h , b e e n k e n 1997). acknowledgements.we thank conservators of herbaria: kra, kram, ktcb, lbl, lod, pozm, szcz, szub, trn, wa, wrsl for their kind help in gathering information about the species as well as dr. anna ronikier, m. sc. marek halama and m. sc. wojciech pusz for their kind help with completing of bibliography and dr. krzysztof kujawa for making the photographs. we also thank anonymous reviewer of our paper. references a n t o n í n v., va g n e r a. 1998. new, rare and less known macromycetes in moravia (czech republic) – iv. acta mus. moraviae. sci. biol. 82: 29–38. b a s c. 1981. een nieuve xylaria voor ons land. coolia 24 (1): 7–10. d e m e u l d e r h. 1984. xylaria oxyacanthae tul., nieuw voor de belgische fungiflora. antwerpse mykologiske kring, mededelingen 84 (2): 33–35. h e t t i c h f., b e e n k e n l. 1997. xylaria oxyacanthae tul. & tul., die „weißdornbeeren-holzkeule”, erstmals in bayern nachgewiesen. mycologia bavarica 2: 61–64. k i r k m. p., c a n n o n p. f., david j. c. & stalpers j. a. (eds). 2001. ainsworth and bisby’s dictionary of fungi. 9-ed. cab. international, wallinford, ss.xi + 655. k r i e g l s t e i n e r g. j. 1983. über neue, seltene, kritische makromyzeten in der bundesrepublik deutschland. z. mykol. 49 (1): 73–106. k r i e g l s t e i n e r g. j. 1993. verbreitungsatlas der großpilze deutschlands (west). bd.ii: schlauchpilze. stuttgart. l æ s s ø e t. 1994. hvor almindelig er tjørne-stødsvamp?. svampe 30: 55. l e r o y p., s u r a u l t j-p. 1995. xylaria oxyacanthae tulasne 1863. observation sur plusieurs mois d’une espèce rarement décrite. doc. mycol. 25 (97): 5–11. m i n t e r d.w. 1986. notes of british species of xylaria. bulletin of the british mycological society 20: 91–93. r e y n d e r s j. 1983. xylaria oxyacanthae en x. carpophila in het amsterdams bos. coolia 26 (3): 60–61. s t o w e l l e. a., r o g e r s j. d. 1983. studies on xylaria oxyacanthae. mycotaxon 17: 433–444. tu l a s n e l. r., tu l a s n e c. 1863. selecta fungorum carpologia. vol. ii. paris. w h a l l e y a. j. s. 1987. xylaria inhabiting fallen fruits. agarica 8 (16): 68–72. w h a l l e y a. 2000. xylaria hill ex schrank. (in:) l. h a n s e n , h. k n u d s e n (eds). nordic macromyce-(in:) l. h a n s e n , h. k n u d s e n (eds). nordic macromycetes. l. ascomycetes. nordsvamp. copenhagen: 249–250. xylaria oxyacanthae tul. et c. tul., gatunek nowy dla polski s t r e s z c z e n i e autorzy prezentują dwa stanowiska nowego dla polski gatunku próchnilca – xylaria oxy­ acanthae. grzyb ten, rozwijający się na owocach crataegus monogyna, został znaleziony po raz pierwszy w zadrzewieniu śródpolnym w turwi (wielkopolska) w roku 2001. drugie stanowisko odkryto w krakowie w parku miejskim w roku 2004. materiał zielnikowy znajduje się w stacji badawczej zbśril pan w turwi i zielniku ib pan w krakowie. fig. 1. xylaria oxyacanthae – locality in turew: a) teleomorphic stage; b) anamorphic stage; c) group of anamorphic stromata. phot. k. kujawa. fig. 2. xylaria oxyacanthae – locality in kraków: a) anamorphic stage; b) anamorfa damaged by armadilidium vulgare; c) teleomorphic stage. phot. d. karasiński. 2014-01-01t11:45:17+0100 polish botanical society new records of smut fungi from venezuela: anthracoidea uleana, sporisorium panici-petrosi and ustilago schroeteriana marcin piątek department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, mpiatek@ib-pan.krakow.pl piątek m.: new records of smut fungi from venezuela: anthracoidea uleana, sporisorium panicipetrosi and ustilago schroeteriana. acta mycol. 43 (2): 153–159, 2008. three smut fungi from venezuela are described, illustrated and discussed based on newly studied collections. ustilago schroeteriana henn. is reported for the first time from the country. anthracoidea uleana (syd. & p. syd.) vánky is confirmed in venezuela on carex longii mack., which is an accessory host for this smut. sporisorium panici-petrosi (syd. & p. syd.) m. piepenbr. is reported on thrasya sp. from a second world locality in northern venezuela, its description is slightly expanded and issues concerning typification of this name and the nomenclature of the host in the type collection are clarified and corrected. key words: anthracoidea, ustilago, sporisorium, smut fungi, neotropics, south america introduction neotropical smut fungi have been recently monographed by piepenbring (2003) who listed 227 species from this part of north and south america. this number seems to be not final, and the author considers that many new species can be discovered in the area as well as numerous species can be found in particular countries as a new national records. indeed several further new species have been subsequently described from different parts of neotropics (vánky 2004, 2005, 2006). venezuela is the sixth largest country of south america, located at the northern coast of the continent, bordering with colombia, brazil, guyana and the caribbean sea. the country has very diverse, tropical landscape, vegetation and vascular plant flora. as in most of neotropical countries, the of smut fungi of venezuela are insufficiently known, and altogether 57 species have been reported from the country till now (sydow and sydow 1916; jackson 1934; dennis 1970; nannfeldt 1977; piepenbring 2003; vánky 2005, 2006, 2007). during ongoing studies focusing on biodiversity of tropical smut fungi, i examined three unidentified collections from venezuela. they belonged to three species, anthracoidea uleana (syd. & p. syd.) vánky, sporisorium panici-petrosi (syd. & p. syd.) m. piepenbr. and ustilago schroeteriana henn. of these, ustilago schroeteriana is new to venezuela, while two remaining acta mycologica vol. 43 (2): 153–159 2008 154 m. piątek species are reported for the second time from the country. these new collections are described, illustrated and discussed in the present contribution. materials and methods dried herbarium specimens were examined by light microscopy (lm) and scanning electron microscopy (sem). for lm, small pieces of sori were mounted in heated lactophenol and examined under a nikon eclipse e600 light microscope. for sem, spores were dusted onto carbon tabs and fixed to an aluminium stub with double-sided transparent tape. the stubs were sputter-coated with carbon using a cressington sputter-coater and viewed with a hitachi s-4700 scanning electron microscope, with a working distance of ca 12−13 mm. sem micrographs were taken in the laboratory of field emission scanning electron microscopy and microanalysis at the institute of geological sciences, jagiellonian university, kraków (poland). results and discussion anthracoidea uleana (syd. & p. syd.) vánky (as “uleiana”), mycotaxon 62: 144 (1997) (fig. 1.) basionym: cintractia uleana syd. & p. syd., ann. mycol. 14: 73 (1916) type: on carex cf. bonplandii kunth (as carex sp.), brazil, roraima, dec. 1909, leg. e. ule 3376 (lectotype: bpi 172260, designated by vánky 1997: 144, isotype: bpi 172261). for further synonyms see piepenbring (2003: 46). sori in ovaries, as black, globose, hard bodies around the ovaries, about 1–1.5 mm in diameter, partly hidden by the scales, composed of agglutinated spores, powdery on the surface. – spores very variable in shape, rounded, bluntly polyhedral, 12–19 × 10–17 μm, olive-brown to dark brown, with 3–5 light, thin-walled, rounded areas that are collapsed and often form characteristic mamillate depressions, which are especially well visible on the sem micrographs; wall uneven, 1–3 μm thick, thickest at the angles, internal swellings and light refractive spots absent, surface as seen by lm finely punctate to verruculose, warts not affecting the spore profile, surface as seen by sem finely verruculose. specimens examined: on carex longii mack. (det. m. piątek), venezuela, state mérida, la carbonera, finca san isidro, alt. ca. 2400 m, moist pasture, 12 jan. 1979, leg. t. ahti 37336, m. lópez figuieras & p. m. jørgensen (h); venezuela, state of lara, w of humocaro bajo, at las sabanetas above los aposentos, alt. ca. 2530 m, wet meadow, 5 feb. 1944, leg. j. a. steyermark (h.u.v. 7638 ex ny ex h). anthracoidea uleana is easy to identify because of unusual spores, which are quite small and possess 3–5 light, thin-walled, rounded areas. these areas are collapsed and often form mamillate depressions, which are especially well visible on the sem micrographs. the host plants of a. uleana belong to carex subgen. vigna sect. ovales. the only other neotropical anthracoidea bref. with similar appearance of spores is anthracoidea altiphila vánky & m. piepenbr., which however differs by some small morphological characters and host plants belonging to carex subgen. carex sect. fecundae (vánky 1994). anthracoidea uleana is one of the four species of anthracoidea known from the neotropics (piepenbring 2003). the small number of species in this floral kingdom 156 m. piątek sporisorium panici-petrosi (syd. & p. syd.) m. piepenbr., flora neotropica 86: 122 (2003) (figs 2 and 4a) basionym: ustilago panici-petrosi syd. & p. syd., ann. mycol. 14: 73 (1916) type: on thrasya petrosa (trin.) chase (=panicum petrosum trin.), venezuela, amazonas, kata, rio cuquenan, dec. 1909, leg. e. ule 3331 (lectotype: s, designated by vánky 2005: 243, erroneously as neotype, icbn art. 9.8). sori usually replacing the whole inflorescence, long, elongated, up to 5 mm wide and up to 10 mm long, sometimes partly concealed by leaf sheaths, at first covered by thick, pale brown peridium, which ruptures irregularly exposing dark brown powdery mass of spores surrounding numerous filiform columellae that are sometimes connected with each other. sori rarely restricted to spikelets, replacing flowers with dusty mass of spores, enclosed by a pale brown peridium and surrounding filiform columellae, partly concealed by glumes. – spores yellowish-brown, on one side lighter coloured, pale yellowish or almost hyaline, globose, subglobose to slightly irregular, 9–13(–15) × (7–)8.5–12 μm; wall ca. 0.5 μm thick, thinner on the lighter coloured side, surface as seen by lm finely verruculose, spore profile smooth, surface as seen by sem moderately verruculose. – sterile cells absent. specimen examined: on thrasya sp., venezuela, estado miranda, caracas, above los palos grandes, s slope of monte naiguata, 1200 m a.s.l., 24 aug. 1958, leg. r.w.g. dennis 2364 [k(m) 134340, together with ustilago schroeteriana]. sporisorium panici-petrosi was so far known only from the type locality in southern venezuela. the collection from northern venezuela reported here represents the second record of this smut (fig. 6). the presence of unusual columellae that are connected with each other as well as characteristic spores that are lighter coloured on one side makes this species easy to identify. according to the descriptions (piepenbring 2003; vánky 2005) of the type collection, the smut destroys whole inflorescences; in the new collection from northern venezuela, however, the sori are restricted only to individual spikelets in some plants while they destroy whole inflorescences in others. the smut was originally described in the genus ustilago (pers.) roussel as u. panici-petrosi syd. & p. syd., but because of the presence of the peridium and columellae, piepenbring (2003) transferred it to sporisorium ehrenb. ex link. vánky (2005) did not accept this proposal, since in his opinion the manner of spore formation in this species is rather of the ustilago-type, and the spores with a thinner wall on one side are typical fig. 5. global distribution of anthracoidea uleana (syd. & p. syd.) vánky with records on carex bonplandii kunth marked by black dots and records on carex longii mack. marked by triangles (modified from piepenbring 2003). new records of smut fungi from venezuela 157 of numerous species of ustilago. however, sporogenesis was analysed in detail only in some species of sporisorium and ustilago (e.g., langdon and fullerton 1975, 1978), and it is not clear whether the way of spore formation is a constant character of taxonomic importance. this is particularly essential in light of recent molecular analyses (stoll et al. 2005) in which sporisorium appeared to be a polyphyletic genus. therefore, the taxonomic value of spore formation in delimiting the genera ustilago and sporisorium should be checked by careful studies of ontogeny and sporogenesis in numerous species, preferably based on freshly collected specimens. in contrast to the statement of vánky (2005), the spores with a thinner wall on one side are known not only in the genus ustilago, but also in several species of sporisorium, for instance in s. clandestinum r.g. shivas, vánky & p. athipunyakom (vánky et al. 2006), s. sphacelatum vánky (vánky 2003), s. wynaadense (sundaram) vánky & r.g. shivas (vánky and shivas 2001). therefore, this character is not useful in delimiting the genera ustilago and sporisorium. in conclusion, i follow the proposal of piepenbring (2003), who placed ustilago panici-petrosi in sporisorium, also because the general appearance of this smut is similar to the species included in this genus. the host plant of sporisorium panici-petrosi was originally named panicum petrosum trin., and for that reason this smut was analysed in the monograph of smut fungi infecting panicum species (vánky 2005). however, panicum petrosum is currently placed in the genus thrasya, as t. petrosa (trin.) chase, which is phylogenetically more closely related to paspalum than to panicum (giussani et al. 2001). the new collection from northern venezuela reported here is on unidentified species of thrasya. in addition, in the same collection there are some plants infected by ustilago schroeteriana, which is a typical paspalum smut. therefore, sporisorium panicipetrosi must be excluded from panicum smuts. the holotype of ustilago panici-petrosi was destroyed in 1943 by berlin fire along with a greater part of the berlin herbarium. fortunately, a duplicate of this collection is preserved in stockholm (herb. s). piepenbring (2003) considered the latter specimen as holotype, which is, however, incorrect as the main collection of the sydows was housed in berlin. vánky (2005) designated the specimen preserved in stockholm (herb. s) as neotype. that denoting is also incorrect, because a neotype can be selected only when all the original material is lost, which is not the case here. therefore, vánky’s denoting is treated as an error (art. 9.8 of icbn), and the specimen in herb. s must be treated as the lectotype of ustilago panici-petrosi. fig. 6. global distribution of sporisorium panici-petrosi (syd. & p. syd.) m. piepenbr. with new locality in northern venezuela marked by triangle (modified from piepenbring 2003). 158 m. piątek ustilago schroeteriana henn., hedwigia 35: 215 (1896) (figs 3 and 4b) type: on paspalum sp., brazil, st. catharina, near itajahy, nov. 1885, leg. e. ule 1615 (holotype: hbg, isotypes: nhes, bpi 166243, 166244, 166245, 194460, h.u.v. 16460). for further synonyms and types see piepenbring (2003: 184) and vánky (2007: 5). sori destroying all spikeletes and peripheral axial tissues of an inflorescence, retaining the shape of affected organs, with shredded remnants of host plant tissue attached to them, spore mass dark brown, powdery. – spores yellow-brown, globose, subglobose to ovoid, variable in size, 13–17(–19) × 12–16(–17) μm; wall even, ca. 1–1.5 μm thick, surface as seen by lm finely verruculose, spore profile finely wavy, surface as seen by sem moderately densely verruculose with punctate warts in the interspaces. specimen examined: on thrasya sp., venezuela, estado miranda, caracas, above los palos grandes, s slope of monte naiguata, 1200 m a.s.l., 24 aug. 1958, leg. r.w.g. dennis 2364 [k(m) 134340, together with sporisorium panici-petrosi]. the morphology of sori and spores of the present collection matches well with those given in descriptions of ustilago schroeteriana (piepenbring 2003; vánky 2007), but the spore sizes are closer to these given by piepenbring (2003): (13–)14–17(–20) × (11–)12–15(–17) μm, than by vánky (2007): 13.5–20 × 12–18.5 μm. the former author pointed out that the spore sizes in different collections of u. schroeteriana are fairly variable, but the most measurements are about 15–17 μm. vánky (2007) in his description gives only extreme values and does not specify which measurements are most common. ustilago schroeteriana is widespread in the neotropics on different species of paspalum, being known from belize, brazil, colombia, costa rica, cuba, granada, honduras, mexico, nicaragua, panama, peru and puerto rico (piepenbring 2003; pérez and minter 2005). the present record is new to venezuela (fig. 7) on the new host plant genus thrasya. the molecular phylogenetic studies (giussani et al. 2001) showed that thrasya is closely related to paspalum and the presence of the same smut fungus on these two host genera additionally supports this close relationship. acknowledgements. i am grateful to my wife jolanta piątek (kraków, poland) for her drawings, to anna łatkiewicz (kraków, poland) for help with the sem pictures, and to the curators of h and k for the loan of specimens. this work was partly supported by the polish ministry of science and higher education (grant no. 2 p04g 019 28 for the years 2005–2008). fig. 7. distribution of ustilago schroeteriana henn. in south america (black dots) with new locality in northern venezuela marked by triangle (modified from piepenbring 2003). new records of smut fungi from venezuela 159 references dennis r. w. g. 1970. fungus flora of venezuela and adjacent countries. kew bull. add. ser. 3: 1–531. giussani l. m., cota-sánchez j. h., zuloaga f. o., kellogg e. a. 2001. a molecular phylogeny of the grass subfamily panicoideae (poaceae) shows multiple origins of c4 photosynthesis. amer. j. bot. 88 (11): 1993–2012. jackson h. s. 1934. ustilaginales. (in:) c. e. chardon, r. a. toro (eds), mycological explorations of venezuela. univ. puerto rico monogr. ser. b., 2: 256–261. langdon r. f. n., fullerton r. a. 1975. sorus ontogeny and sporogenesis in some smut fungi. austral. j. bot. 23: 915–930. langdon r. f. n., fullerton r. a. 1978. the genus sphacelotheca (ustilaginales): criteria for its delimitation and the consequences thereof. mycotaxon 6: 421–456. nannfeldt j. a. 1977. the species of anthracoidea (ustilaginales) on carex subgen. vignea with special regard to the nordic species. bot. notiser 130: 351–375. pérez j. m., minter d. w. 2005. ustilago schroeteriana. [descriptions of fungi and bacteria]. imi descriptions of fungi and bacteria, no. 164, sheet 1640. piepenbring m. 2003. smut fungi (ustilaginomycetes p.p. and microbotryales, basidiomycota). flora neotropica 86: iv+1–291. piepenbring m. 2006. neue ergebnisse zur diversität und ökologie von brandpilzen in panama. z. mykol. 72(2): 89–100. stoll m., begerow d., oberwinkler f. 2005. molecular phylogeny of ustilago, sporisorium, and related taxa based on combined analyses of rdna sequences. mycol. res. 109: 342–356. sydow h., sydow p. 1916. fungi amazonici a cl. e. ule lecti. ann. mycol. 14: 65–97. vánky k. 1994. taxonomical studies on ustilaginales. xi. mycotaxon 51: 153–174. vánky k . 1997. taxonomical studies on ustilaginales. xv. mycotaxon 62: 127–150. vánky k. 2003. taxonomical studies on ustilaginales. xxiii. mycotaxon 85: 1–65. vánky k. 2004. new smut fungi (ustilaginomycetes) from mexico, and the genus lundquistia. fungal divers. 17: 159–190. vánky k. 2005. taxonomic studies on ustilaginomycetes – 25. mycotaxon 91: 217–272. vánky k. 2006. taxonomic studies on ustilaginomycetes – 26. mycotaxon 95: 1–65. vánky k. 2007. taxonomic studies on ustilaginomycetes – 27. mycotaxon 99: 1–70. vánky k., shivas r. g. 2001. smut fungi (ustilaginomycetes) of sorghum (gramineae) with special regard to australasia. mycotaxon 80: 339–353. vánky k., shivas r. g., athipunyakom p. 2006. new smut fungi (ustilaginomycetes) from thailand. mycol. balcanica 3(2–3): 107–118. nowe stwierdzenia grzybów głowniowych w wenezueli: anthracoidea uleana, sporisorium panici-petrosi i ustilago schroeteriana s t r e s z c z e n i e w pracy zaprezentowano trzy gatunki grzybów głowniowych z wenezueli, stwierdzone podczas rutynowego oznaczania niezidentyfikowanych materiałów pochodzących z obszarów tropikalnych. ustilago schroeteriana henn. jest nowy dla tego kraju. anthracoidea uleana (syd. & p. syd.) vánky jest podany z drugiego stanowiska w wenezueli. sporisorium panici-petrosi (syd. & p. syd.) m. piepenbr. jest podany z drugiego stanowiska na świecie w północnej wenezueli. w pracy rozszerzono nieco opis tego gatunku o nowe cechy diagnostyczne i wyjaśniono lub poprawiono niektóre błędne informacje dotyczące jego typizacji i nomenklatury rośliny żywicielskiej. 2014-01-01t11:47:56+0100 polish botanical society 2014-08-28t14:43:59+0200 polish botanical society persistence of ectomycorrhizas by thelephora terrestris on outplanted scots pine seedlings dorota hilszczańska and zbigniew sierota forest research institute bitwy warszawskiej 1920 r. nr 3, pl 00 973 warszawa d.hilszczanska@ibles.waw.pl, z.sierota@ibles.waw.pl h i l s z c z a ń s k a d., s i e r o t a z.: persistence of ectomycorrhizas by thelephora terrestris on outplanted scots pine seedlings.acta mycol. 41 (2): 313 318, 2006. thelephora terrestris (erhr.) fr. is a very common ectomycorrhizal symbiont (ecm) in conifer trees, however the role of this ubiquitous fungus in nurseries and scots pine plantations is still unknown. it is described as tolerant of high nitrogen availability and therefore was taken into consideration as an important ecm partner of seedlings, particularly after replanting on post agricultural land. in laboratory the seedlings of scots pine (pinus sylvestris l.) were inoculated with t. terrestris (tt/ibl/2) or not inoculated (control) and grown in containers in two different regimes of nitrogen fertilization (4g n x kg 1 and 6 g n x kg 1). next year these seedlings were outplanted in post agricultural land and 6 months later, the number and identity of some mycorrhizas were studied. it was found, that mycorrhizal abundance was higher in the inoculated treatments than in non inoculated ones. pcr rflp analysis confirmed share of two different isolates of thelephora engaged in mycorrhizal symbiosis. part of mycorrhizas had the same pattern of rflp as the isolate used to inoculation. similar results were obtained in second year of experimental study in the field what confirmed the persistence of artificially introduced t. terrestris in post agricultural soil as an important component of the ecm community. key words: thelephora terrestris, scots pine seedlings, inoculation, ectomycorrhiza, post agricultural land, pcr, rflp introduction the root systems of most forest trees, including pines, form ectomycorrhizas (ecm) with various fungi. ecm colonization rate and species composition play an important role in the stability of forest ecosystems (h a s e l w a n d t e r , b o w e n 1996), especially on disturbed soils (m u l l i n s et al. 1989). successful afforestation requires the early capture of site resources by tree seedlings. early growth assures space, a continuing resource supply and vigour to resist pests, pathogens and climatic stress (p e r r y et al. 1987). acta mycologica vol. 41 (2): 313-318 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 314 d. hilszczańska and z. sierota in order to improve adaptation of pine seedlings destined to non-forest soils the artificial inoculation has been successfully used (b e r r y , m a r x 1978; c a s t e l l a n o 1996). according to the national afforestation programme estimated area of abandoned lands that should be afforested is about 1.5 millions ha (g r z y w a c z 2000). most frequent morphotypes identified on such sites were pisolithus tinctorius, scleroderma citrinum, thelephora terrestris, suillus sp., tuber sp., tomentella sp. (d a n i e l s o n 1991; h i l s z c z a ń s k a 2002). the natural variability makes it necessary to develop selection strategies for ecm fungi as inocula in forest nurseries. the majority of screening programmes search for isolates that increase nutrient uptake and plant growth. recent years have seen an increased awareness of the additional benefits of a fungal partner in the root system. hence, different criteria may be used to select inoculant mycorrhizal fungi unrelated to ability to increase growth rates or nutrient uptake (d o d d , t h o m s o n 1994). b o w e n (1994) claims that fungi used as inocula should be related and appropriate to soil conditions such as humidity, texture, chemistry and nutrient availability. t. terrestis is a very common ectomycorrhizal symbiont and occurs on a wide variety of soils, including non-fertilised and highly fertilised, mineral or peaty soils, and in dry or wet conditions (c o l p a e r t 1999). in presented study t. terrestris mycelium was used as inoculum for seedlings which were grown in control conditions in the laboratory. after inoculation two different nutrient supply were applied and next year later, 1-year old seedlings were outplanted on post-agricultural land. the aim of this work was to estimate the presence and share of t. terrestris mycorrhizas. materials and methods for the laboratory study, the selected seeds of p. sylvestris were taken from gene bank in kostrzyca, poland and surface sterilised for 15 min. in 30 % hydrogen peroxide. the seeds were sown to containers (root trainer, v 123 cm3) containing a previously autoclaved mixture of peat moss and vermiculite (2:1, v/v). nitrogen was the growth limiting element and consisted of 47.4 % ammonium and 52, 6 % nitrate. in one variant used, each plant was fertilized with 0, 2g n per plant, that gives 4g n x kg-1 of soil (low n), and in second variant each plant was fertilized with 0,3 g n per x kg-1 of soil plant 6g x kg-1 of soil (high n). the phkcl of soil was 4.1 the experiment was carried out during 1 year in a growth chamber with a day/night rhythm of 18/6 h and 22/15ºc at least 70% relative air humidity. inoculation of seedlings was done 28 days after sowing, with blended mycelium of thelephora terrestris (erhr.) fr. (tt/ibl/2, isolated from the fruitbody); each seedling received 5 ml of mycelium suspended in deionised water (0.025 g mycelium per plant). seedlings were grown in four treatments: 1high n and inoculation, 2low n and inoculation, 3high n without inoculation, 4low n without inoculation (h i l s z c z a ń s k a , s i e r o t a 2006). six months after inoculation, 15 seedlings from each treatment were selected randomly for determination of the degree of mycorrhizal colonisation. next year, at the beginning of spring 2004 seedlings were outplanted on post agricultural land in jabłonna forest district. within the area, mean annual precipitation is about 522.8 mm and average temperature during the growth season is ca. 8.4ºc. the land had been abandoned for over 1year prior to our study. the site is characterised by sandy soil, corresponding to vaccinio-myrtilliosa forest type. planting was carried out persistence of ectomycorrhizas 315 in the scheme of randomised block design with four treatments as previously. soil samples (cores were taken to a depth of 20 cm at 5 different location) were collected in april 2004. soil phkcl and the basic nutrients were determined (tab. 1). after the first growing season (2004) and next after the second year (2005) samples of plants (15 plants from each treatment) were taken in order to assess the presence of t. terrestris mycorrhizas and the composition of the ecm communities. prior to investigation, each root systems were excised from the stems and washed in tap water. mycorrhizal tips were counted and morphotyped as described by a g e r e r (1987-1997), i n g l e b y et al. (1990) and p a r l a d e et al. (1996). the morphotypes which did not match any of those presented in cited material were classed as unidentified. in order to confirm that morphotyping was well done, 22 mycorrhizal tips from all treatments were taken to dna analysis with pcr rflp method, conducted in the institute of biology and biochemistry. the fungal specific primer its1-f (g a r d e s , b r u n s 1993) and universal primer its 4 (w h i t e et al. 1990) have been used for amplification by pcr. if only one dna band was present per sample (confirming that all dna came from one source only), the product was used for rflp analysis. digestions with the restriction enzymes hinf i, mbo i and taq i were performed using 18 ul dna from amplifications of mycorrhizal origin. analysis of the restriction patterns was made on 2,4 agarose gels which were stained with ethidium bromide. the variability of mycorrhizas was assessed by analysis of variance (anova), significant differences of means were compared by lsd (p<0.05) (statsoft, inc. (2005). statistica (data analysis software system, version 7,1. www.statsoft.com) 1998). results after the first season, irrespective of nitrogen amount, higher number of mycorrhizas possessed the inoculated seedlings than non-inoculated ones (fig. 1a). at inoculated treatments number of mycorrhizas did not differ significantly between low n and high n: 659 and 728, respectively. at non-inoculated treatments the number of mycorrhizas was significantly lower; in low n 489 and in high n 328. the differences between inoculated and non-inoculated seedlings were also statistically significant. after the first growing season, the mycorrhizas at inoculated treatment with high amount of n were mostly formed by t. terrestris (tab. 2), while at treatment with low n mycorrhizas formed by hebeloma sp. had the highest number. moreover, the last mycorrhizas were dominant morphotype on non-inoculated seedlings, irrespecta b l e 1 mean content of minerals in the soil 20 cm deep (g x kg 1) and ph c n ca mg k p s al phkcl 8.3 0.85 1.17 0.67 0.75 0.63 0.10 5.55 4.48 316 d. hilszczańska and z. sierota tive of n level. at all treatments percentage of mycorrhizas formed by amanita sp. varied from 5 to 10 %. mycorrhizas formed by thelephora sp. at non-inoculated treatment belonged to a strain that we called as thelephora ii – indigenous in that soil. it has been confirmed by pcr rflp analysis (tab. 3). thelephora i was the same strain that had been used to inoculation. assessment of the accurate number of mycorrhizas formed by both strains was not possible due to lack of morphological differences. among all fig. 1. number of mycorrhizas on scot pine seedlings: a first growing season, b the second growing season. statistically significant differences between the treatments (1 high n and in oculation, 2 low n and inoculation, 3 high n without inoculation, 4 low n without inocula tion) are designated by different letters. error bars indicate standard error of the mean. ta b l e 2 frequency of mycorrhizal morphotypes on root tips of pinus sylvestris seedlings under different treatments in 2004 and 2005 morphotypes year 2004 year 2005 inoculated non inoculated inoculated non inoculated high n low n high n low n high n low n high n low n amanita sp. hebeloma sp. suillus sp. thelephora terrestris unidentified 10.0 10.0 75.0 5.0 5.0 60.0 30.0 5.0 4.0 80.0 10.0 6.0 8.0 75.0 15.0 2.0 10.0 30.0 60.0 25.0 5.0 70.0 10 5.0 35.0 50.0 5 25.0 5.0 65.0 ta b l e 3 approximate restriction fragment sizes (bp) of the amplified fungal its region of t. terrestris mycorrhizas fungus its hinf i mbo i tag i thelephora terrestris (pure culture) 680 315 240 103 360 225 60 295 270 thelephora (i) 663 318 242 105 365 226 58 297 270 65 thelephora (ii) 718 228 208 134 115 245 236 155 67 295 224 108 98 persistence of ectomycorrhizas 317 mycorrhizas analysed by pcr rflp method, strain thelephora i was found in 36 % of samples and strain thelephora ii in 64 % of samples. after the second growing season, irrespective of inoculation or fertilization, all seedlings possessed higher number of mycorrhizas than after the first growing season. the highest number of mycorrhizas was observed at inoculated treatment with high amount of n and the lowest one at noninoculated treatment with high amount of n (fig. 1b). mycorrhizas of t. terrestris were found on seedlings at all treatments, but the highest share was found at treatment inoculated with low n (tab. 2). in mycorrhizal structure emerged mycorrhizas of suillus sp. that had similar percentage at high level of n in case inoculated and non-inoculated treatments. the highest percentage of those mycorrhizas was noticed at non-inoculated treatment with high n. according to pcr rflp analysis (which was successful in case of 30 mycorrhizas) the percentage of mycorrhizas formed by thelephora i was about 30%, whereas by thelephora ii was 70%. discussion after two growing seasons following plantation establishment, the scots pine seedlings inoculated with t. terrestris showed significantly higher number of mycorrhizas than non-inoculated seedlings (fig. 1a and b). the morphotypes of t. terrestris were also found on non-inoculated seedlings, it was clearly confirmed, that these ones were formed by indigenous strain of t. terrestris colonising only the limited part of roots, however. the other observed mycorrhizas were similar to those commonly observed in forest nurseries (m e n k i s et al. 2005). the dominant mycorrhizas on non-inoculated seedlings and seedlings that were grown at low n fertilization belonged to hebeloma sp. after second growing season the number of these mycorrhizas in root system was rather small. according to g u i d o t et al. (2003) some fungi as h. cylindrosporum were replaced by others even after one growing season and do not necessarily contribute to next generation of mycorrhizas. similar results described m e n k i s at al. (2005).the results indicate that i) the success of mycorrhizal inoculation in the field largely depends on fungal species, both used in mycorrhization and existing in the local soil, and ii) the ecological conditions of the soil will drive mycorrhizal colonisation at a given site (m c a f e e , f o r t i n 1985). this site was suitable for t. terrestris since after second season the mycorrhizas were the dominant morphotype. even if inoculated strain was in small portion, compare to the indigenous one, it was still present. this might suggested that t. terrestris is a strong competitor or that can thrive in species-poor communities. references a g e r e r r. 1987 1997. colour atlas of ectomycorrhizae. einhorn verlag, schwabisch gmünd. b e r r y c. r., m a r x d. h. 1978. survival and growth of pine hybrid seedlings with pisolithus ectomycor rhizae on coil spoil in alabama and tennessee. j. environ. qual. 11: 709 715. b o w e n g. d. 1994. the ecology of ectomycorrhiza formation and functioning. plant and soil 159: 61 67. c a s t e l l a n o m. a. 1996. outplanting performance of mycorrhizal inoculated seedlings. (in:) k. g. m u k e r j i (ed.). concepts in mycorrhizal research. handbook of vegetation science kluwer, dor drecht 19 (2): 223 301. 318 d. hilszczańska and z. sierota c o l p a e r t j. v. 1999. thelephora. (in:) j. w.g c a i r n e y , s. m c h a m b e r s (eds). ectomycorrhizal fungi. key genera in profile. springer. d a n i e l s o n r. m. 1991. temporal changes and effects of amendments on the occurrence of sheating (ecto )mycorrhizae of conifers growing in oil sands tailings and coal spoil. agric. ecosyst. environ. 35: 261 281. d o d d j .c., t h o m s o n b.d. 1994. the screening and selection of inoculant arbuscular mycorrhizal and ectomycorrhizal fungi. plant and soil 159: 149 158. g a r d e s m., b r u n s t. 1993. its primers with enhanced specifity for basidiomycetes application to the identification of mycorrhizas and rusts. mol. ecol., 2:113 118. g r z y w a c z . a. 2000. stan i potrzeby w zakresie mikoryzacji sadzonek drzew leśnych w polsce na tle pro dukcji szkółkarskiej oraz zadań w zakresie zwiększenia lesistości kraju. (in:) m. r u d a w s k a (ed.). ektomikoryza, jej znaczenie i zastosowanie w leśnictwie, instytut dendrologii pan, kórnik. g u i d o t a., d e b a u d j. c., e f f o s s e a., m a r m e i s s e r. 2003. below ground distribution and per sistence of an ectomycorrhizal fungus. new phytol. 161: 539 547. h a s e l w a n d t e r k., b o w e n g. d. 1996. mycorrhizal relations in trees for agroforestry and land reha bilitation. for. ecol. manage. 81: 1 17. h i l s z c z a ń s k a d. 2002. mycorrhizal fungi in scots pine cultures after seedlings out planting on post agricultural lands. folia forestalia polonica, series a forestry 44: 97 102. h i l s z c z a ń s k a d., s i e r o t a z. 2006. the role of thelephora terrestris fungus in mycorrhization on scots pine (pinus sylvestris l.) seedlings i. laboratory study. sylwan 1: 40 47. i n g l e b y k., m a s o n p.a., l a s t f.t., f l e m i n g l.v. 1990. identification of ectomycorrhizas. ite re search publication no.5. institute of terrestrial ecology. london:hmso. m c a f e e b. j., f o r t i n j. a. 1985. competitive interactions of ectomycorrhizal mycobionts under field conditions. can. j. bot. 64: 848 852. m e n k i s a. 2005. root associated fungi of conifer seedlings and their role in afforestation of agricultural land. swedish university of agricultural sciences, doctoral thesis 106, uppsala. p a r l a d e j., a l v a r e z i. f., p e r a j. 1996. inoculation of containerized pseudotsuga menziesii and pinus pinaster seedlings with spores of five species of ectomycorrhizal fungi. mycorrhiza 6: 237 245. p e r r y d. a., m o l i n a r., a m a r a n t h u s m. p. 1987. mycorrhizae, mycorrhizpspheres, and reforesta tion: current knowledge and research needs. can. j. for. res. 17:929 940. w h i t e t. j., b r u n s t., l e e s., ta y l o r j. 1990. amplification and direct sequencing of fungal ribo somal rna genes for phylogenetics. (in:) m. a. i n n i s , d. h. g e fl a n d , j. s n i n s k y , t. j. w h i t e (eds). pcr protocols. a guide to methods and amplifications: 315 322. academic press. trwałość ektomikoryz thelephora terrestris u sadzonek sosny w uprawie s t r e s z c z e n i e sadzonki sosny zwyczajnej pinus sylvestris l., mikoryzowane grzybnią thelephora terrestris i nie mikoryzowane (kontrola) hodowano w laboratorium w dwóch wariantach nawożenia (4g n x kg 1 and 6 g n x kg 1) a po roku wysadzono na gruncie porolnym. w ciągu dwóch sezonów wegetacyjnych badano stan ilościowy i jakościowy mikoryz oraz określono tożsamość partnera grzybowego. wyższą ogólną liczbą mikoryz charakteryzowały się sadzonki sztucznie miko ryzowane. analiza dna grzybowego przeprowadzona metodą pcr rflp wykazała, że na badanych korzeniach mikoryzy t. terrestris tworzone były przez dwa izolaty thelephora; izolat użyty do inokulacji siewek (thelephora i) i izolat obecny w glebie (thelephora ii). mikoryzy tworzone z udziałem inokulowanej grzybni obecne były na korzeniach jeszcze w dwa lata po wysadzeniu sadzonek na gruncie porolnym i stanowiły 30% udziału, podczas gdy mikoryzy autochtoniczne thelephora ii 70%. 2014-01-01t11:44:49+0100 polish botanical society 2014-08-28t17:48:52+0200 polish botanical society 266 polish mycological society the polish mycological society (pms) was established on 5 november 2011, in łódź. the aims of the society are to develop, coordinate and support mycological studies, and to popularize mycology in its widest sense among the people in poland, in cooperation with mycologists and mycological societies worldwide. as required by polish law, the society was registered (entry 00004 22681) on 23 june 2012 in the national court register of the city of warsaw district court. contact information for the society is as follows. website: www.ptmyk.pl e-mail: polskietowarzystwomykologiczne@gmail.com. postal address (under its polish name): polskie towarzystwo mykologiczne (ptmyk), al. ujazdowskie 4, pl-00-478 warszawa, poland. the background for establishing the pms was both general and local. historically, mycology was represented only through sections of other societies including the polish botanical society, polish hygienic society, polish dermatological society, and polish society of building mycology. interest in scientific study and practical use of fungi increased after recognized them as a separate kingdom within eukaryota. as a result the influence of mycology, and its connection with and presence in many areas of knowledge became more evident. this led mycologists to realize that their science faced structural issues: coverage of mycology lacked focus. the increase in scientific mycology had a social counterpart with the remarkable rise in numbers of amateurs interested in fungi, particularly rare and little known species (the website www.grzyby.pl is excellent example of their work). their research, observations and documented collections suggested that enhanced contact and collaboration between amateurs and professionals would be good for mycology. there was a clear need to integrate these different elements into a single forum for exchanging ideas and information. external conditions also became more favourable, with the appearance of new scientific infrastructure at continental and global levels. in 1985 the european council for conservation of fungi was established, and that was followed in 2003 by formation of the european mycological association. polish mycologists participating in these initiatives became aware that they were among a minority of europeans who lacked their own national mycological society. the gestation period of the new mycological society was long: discussions about the need for a national mycological society and the form it should take lasted several decades. it all started when the mycological section of the polish botanical society was set up in 1957. this gathered together experimental mycologists and researchers in poland, and was headed by professor alina skirgiełło for 50 years. in september 2001, at a forum of this mycological section of the polish botanical society, a proposal to establish a polish mycological society was submitted by maria ławrynowicz, on behalf of the board of that section. the resulting discussions supporting and motivating the idea lasted ten years and spanned several meetings (2004, 2007, 2009, 2010, 2011). they resulted in preparation of a constitution and, finally, in the formation of the pms on 5 november 2011, during a special nationwide polish mycological convention held under the auspices of the european mycological association in the presence of david minter, its president and stephanos diamandis, its past vice-president. all the necessary documentation was unanimously accepted by that meeting, and the establishment of the pms enthusiastically approved, by the 34 founder members (fig. 1). this historic event was the culmination of long efforts by three generations of polish mycologists. the meeting appointed an initial executive committee as follows: dr marta wrzosek as president, dr hab. janusz łuszczyński as vice-president, dr małgorzata ruszkiewicz-michalska as secretary, dr julia (budziszewska) pawłowska as treasurer. maria ławrynowicz and marta wrzosek 2014-01-02t12:17:46+0100 polish botanical society 2014-08-25t21:38:54+0200 polish botanical society 2014-08-25t14:56:23+0200 polish botanical society 2014-08-20t22:12:35+0200 polish botanical society saccharomyces cerevisiae in the respiratory system, digestive system and on the skin in humans maria dynowska, małgorzata rosłan1 and katarzyna góralska department of mycology, university of warmia and mazury in olsztyn oczapowskiego 1a, pl 10 957 olsztyn, dynow@uwm.edu.pl 1laboratory of bacteriology of independent public centre for tuberculosis and lung diseases jagiellońska 78, pl 10 357 olsztyn d y n o w s k a m . , r o s ł a n m . , g ó r a l s k a k .: saccharomyces cerevisiae in the respiratory system, digestive system and on the skin in humans. acta mycol. 41(1): 139 144, 2006. the results of observations on the occurrence of s. cerevisiae in the respiratory and digestive systems and on the skin in hospitalised individuals, mostly belonging to so called risk groups of fungal infections, conducted over a period of five years (1999 2004), are described and discussed. the participation of s. cerevisiae in the mycocoenoses studied was 11.04% of all the recorded fungi. a significant increase in its prevalence, especially on the skin, was noticed. the fungus was mostly isolated with fungi of the genera candida and trichosporon, as well as separately in the last two years of the study. key words: s. cerevisiae, ontocoenoses, prevalence introduction true yeasts of the genus saccharomyces are some of the numerous fungi that may colonise the oral cavity and the digestive tract in humans commensally and asymptomatically (k u r n a t o w s k a 1995). s. cerevisiae, one of the many species of this genus, is mostly associated with fermentation processes or the production of some vitamins (b1, b2, pp, pantothenic acid) and less frequently with pathogenic processes. biologists know that yeasts are an excellent object of study in cytology, enzymology, growth and development physiology, dimorphism and genetics (d y n o w s k a 1995). relatively little, however, is known about yeasts as causal agents of mycoses despite an increasing number of reports from medical mycologists on the expanding range of aetiological factors of diseases caused by yeasts (d y n o w s k a , b i e d u n k i e w i c z 2001). only few list s. cerevisiae as a potential anthropopathogen belonging to opportunistic fungi. long-term observations on human-related fungi, both in healthy and compromised individuals, based on routine mycological diagnostics using clinical material, show that the occurrence frequency of s. cerevisiae in various human ontocoenoses acta mycologica vol. 41 (1): 139-144 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 140 m. dynowska et al. deserves attention (d y n o w s k a , e j d y s , k i s i c k a 2004). therefore, its occurrence in positive mycological results of the laboratory of bacteriology at the independent public centre for tuberculosis and lung diseases, olsztyn, was analysed over a period of five years. material and methods smears from various body sites, including bedsores and post-operative wounds, smears from the oral cavity, throat, large intestine and anus as well as sputum and bronchoscopic fluid constituted the study material. fungi were incubated according to general guidelines recommended for diagnostic laboratories: macrocultures – sabouraud agar, microcultures – nickerson agar, incubation at 37˚c for 48-72 hours (k u r n a t o w s k a 1995; p a w l i k , m a c u r a 1998). a total of 2 880 samples in which any fungi were recorded were examined over a period of five years (1999-2004). samples were collected from 1 420 men and 1 460 women who constituted a group of oncological patients and patients suffering from respiratory system disorders, including suspected tuberculosis. yeasts mostly accompanied numerous yeast-like fungi. samples containing s. cerevisiae were additionally tested in sabouraud broth (microscopic sediment analysis) and fermentation and sugar assimilation abilities were assessed. the production of asci and pseudohyphae was observed on gorodkow agar and a cornmeal substrate, respectively (k u r n a t o w s k a 1995). fungi were determined using the following keys: b a r n e t t , p a y n e and ya r r o w (1990) and k r e g e r v a n r i j (1984). results a total of 318 isolates of s. cerevisiae, 145 in men and 173 in women, were obtained from the patients’ respiratory and digestive systems and skin, which constitutes 11.04% of all recorded fungi (tab. 1). yeasts accompanied fungi of the genera candida and trichosporon in the vast majority of cases. s. cerevisiae was observed separately in the last two years of the study (2003 and 2004). in these cases, growth was abundant, and the entire field of vision in direct microscopic slides was covered by vegetative cells and intensively budding cells. the digestive system, followed by the respiratory system, was the ontocoenosis colonised by s. cerevisiae most often. the fungus was recorded on the skin very rarely between 1999 and 2002, and its frequency was comparable to that in other ontocoenoses between 2003 and 2004. while yeasts occurred in the respiratory and digestive systems in men and women with a similar frequency (with a slight shift towards women), they were recorded almost twice as often on the skin in women (tab. 1). they were usually isolated in summer and autumn, and rarely in spring and winter. a significant increase in the prevalence of the species studied was observed in the study period and it went from 1.90% in 1999 to 3.53% in 2005. saccharomyces cerevisiae in the respiratory 141 ta b l e 1 number of s. cerevisiae isolates in the study material in women (w) and men (m) in different seasons of the year (1999 2004) year season ontocoenosis total total % (in relation to the total number, i.e. 2880) respiratory system digestive system skin w m w m w m w m 1999/2000 sp 1 3 2 2 6 2 45 1.90 su 2 1 6 4 4 2 12 7 a 1 1 6 4 1 1 8 6 2000/2001 w 1 2 1 3 1 37 1.21 sp 2 1 2 2 4 3 su 3 2 4 2 2 2 9 6 a 3 2 3 1 1 1 7 4 2001/2002 w 1 1 1 1 3 1 60 2.08 sp 2 1 4 3 2 8 4 su 5 3 8 5 1 14 8 a 3 2 7 5 2 12 7 2002/2003 w 1 1 2 2 1 4 3 74 2.22 sp 2 2 3 3 2 7 5 su 4 6 (2) 5 6 (2) 4 (1) 4 (1) 13 16 a 4 (2) 4 (2) 5 (2) 6 (3) 4 (1) 3 (1) 13 13 2003/2004 w 2 1 2 2 4 3 102 3.53 sp 2 4 3 3 3 (1) 1 8 8 su 8 (3) 10 (2) 6 (2) 6 (2) 4 (2) 4 (2) 18 20 a 8 (2) 10 (2) 3 6 (1) 4 (2) 4 (2) 15 20 w 2 (2) 5 (2) 1 2 (1) 2 1 5 8 total 57 56 76 66 40 43 173 145 318 11.04 113 142 63 318 % (in relation to the total number, i.e. 2880) 3.89 4.97 2.18 11.04 (1) number of s. cerevisiae isolates occurring separately; sp spring, su summer, a autumn, w winter 142 m. dynowska et al. discussion s. cerevisiae is a typical representative of f. saccharomycetaceae, o. endomycetales which combines features of zygomycetes and ascomycetes. dicaryophase, ascocarps and ascogenous hyphae are absent in yeasts, and asci are produced directly from the zygote. s. cerevisiae cells are characterised by a great size differentiation depending on the strain: 4.5 – 10.5 x 7.0 – 21.0 μm; 3.5 – 8.0 x 5.0 – 11.5 (17.5) μm; 2.5 – 7.0 x 11.0 – 18.5 μm. the diameter of single cells of some strains was up to 30μm. the chemical composition and the organisation of the structure of the cell wall in yeasts diverge from those of most fungi. it consists of cell dry mass in 1920%; polysaccharides (mostly glucans and mannans) constitute 80–90% while chitin only ca. 10%. the thallus of s. cerevisiae is small and reproduces by budding, leaving scars, with blastospores. it may be haploid or diploid in the vegetative stage. it may elongate into pseudohyphae, sometimes breaking down into blastospores, in microaerophilous conditions or when trophic conditions are disturbed. however, budding is mostly the only form of growth in s. cerevisiae: intensive multiplication and an increase in the number of vegetative cells take place as the biomass increases. it is of great significance for the rate of medium colonisation, including human tissue (d y n o w s k a 1995). yeasts of two genera, saccharomyces and hansenula, have been isolated from the human body, and only saccharomyces occurred asymptomatically. apart from s. cerevisiae, s. italicus casteli and s. rouxii boutroux have been observed. they have mostly been recorded in the contents of the oral cavity and in faeces, less frequently in other biological material (k u r n a t o w s k a 1995). both genera can cause systemic yeast infections. saccharomycoses have been recorded in the oral cavity, stomach, intestines, reproductive organs, respiratory system (bronchi and lungs), in the external ear and on the skin. hansenula infections have been recorded in the blood, central nervous system, endocardium and lymph nodes (k u r n a t o w s k a 1995). a relatively low percentage of s. cerevisiae in the study material should not divert attention from the species’ pathogenic abilities, especially because the number of its records has been growing and the fungus is beginning to occur on its own. its presence on the skin, indicative of the acquisition of keratinophilous properties, is particularly dangerous. trichosporon beigelii is an example of species that began colonisation of human tissues from the skin and its products (wa r n o c k , r i c h a r d s o n 1991). it initially entered the urinary and genitourinary systems and then the respiratory system (d y n o w s k a 1995). apart from candida albicans, it is currently a frequent aetiological factor of mycoses of various human ontocoenoses (d y n o w s k a 1996). while isolating s. cerevisiae from the oral cavity, throat or the digestive system, especially in summer when large amounts of fruit are consumed, does not have to indicate initial stages of mycosis, recording isolates in other biological material raises certain suspicions (d y n o w s k a , h o ł o w e ń c z a k , f i e d o r o w i c z 1997). the presence of yeasts in bronchoscopic material and in the large intestine shows the state of potential threat. yeasts usually accompanied yeast-like fungi characterised by high pathogenicity: c. albicans, c. glabrata, c. tropicalis, t. beigelii. these species were most probably the direct cause of pathogenic changes while the yeasts constituted a secondary additional factor, which does not exclude the reverse situation. this remark seems saccharomyces cerevisiae in the respiratory 143 particularly significant as a gradual decrease in immunity to infections caused by various fungi accompanying man and an increase in the expansive nature of a number of opportunistic forms, sometimes believed to be accidental contamination of clinical material or, even worse, a physiological element, have been observed (s e n e t , r o b e r t 1995). due to high enzymatic activity, yeasts that had been ingested or inhaled could begin destructive processes and paved the way for yeast-like fungi. the danger posed by s. cerevisiae is mostly caused by its ability to grow and develop at the temperature of the human body, at the decreased oxidation and reduction potential that prevails in diseased tissue. it ecophysiology is within the definition of fungi potentially pathogenic to people (r i c h a r d s o n , wa r n o c k 1995). yeasts are very common saprotrophes, and they naturally occur on fruit surfaces, flower nectaries, cracked tree bark and plant secretions rich in carbohydrates, eutrophied water reservoirs abundant in organic matter, and soil (d y n o w s k a 1995). all fungi colonising the human body should be carefully monitored, especially as susceptibility to fungal infections is common and affects individuals of any age, both healthy and compromised. since the range of factors predisposing to fungal infections is expanding, it is impossible to determine when a symbiotic commensal transforms into a parasite. conclusions s. cerevisiae belongs to fungi that have increasingly been colonising various human ontocoenoses, including the skin. the results of clinical studies in which s. cerevisiae occurs should be analysed in depth as its enzymatic activity and expansive behaviour in relation to human tissues is growing. references b a r n e t t j . a . , p a y n e r . w. , ya r r o w d . 1990. yeast: characteristic and identification. cam brige univ. press. d y n o w s k a m . 1995. drożdże i grzyby drożdżopodobne jako czynniki patogenne i bioindykatory eko systemów wodnych. studia i materiały wsp olsztyn 77: 1 83. d y n o w s k a m . 1996. trichosporon species isolated from human respiratory system. acta mycol. 31 (2): 137 141. d y n o w s k a m . , b i e d u n k i e w i c z a . 2001. grzyby chorobotwórcze o wzrastającej ekspansywności. wiad. parazytol. 47 (4): 609 613. d y n o w s k a m . , e j d y s e . , k i s i c k a i . 2004. susceptibility to antifungal agents of yeast like fungi and yeast isolated from people with multilocal infections. mikol. lek. 11 (1): 15 19. d y n o w s k a m . , h o ł o w e ń c z a k a . , f i e d o r o w i c z g . 1997. drożdże i grzyby drożdżopodobne izolowane z wybranych owoców i nasion jako mało znane czynniki patogenne. (in:) konferencja naukowa promocyjna „lepsza żywność” (iv). art. olsztyn kortowo: 52 55. k u r n a t o w s k a a . 1995. wybrane zagadnienia z mikologii medycznej. wyd. ii. promedi, łódź. k r e g e r v a n r i j n . j . w. 1984. the yeasts: a taxonomic study. third revision and enlarged edition. els. sci. publ. amsterdam. p a w l i k b . , m a c u r a a . b . 1998. diagnostyka laboratoryjna w mikologii. (in:) e. b a r a n (ed.). zarys mikologii lekarskiej. volumen wrocław: 541 572. r i c h a r d s o n m . d . , wa r n o c k d . w. 1995. grzybice rozpoznanie i leczenie. springer. pwn, warszawa. s e n e t j . m . , r o b e r t r . 1995. physiopathologie des candidoses. j. mycol. med. 5: 145 166. wa r n o c k d . w. , r i c h a r d s o n m . d . 1991. fungal infection in the compromised patients. chi chester john viley. 144 m. dynowska et al. saccharomyces cerevisiae w układzie oddechowym, pokarmowym i na skórze człowieka s t r e s z c z e n i e przeanalizowano (1999 2004) udział s. cerevisiae w dodatnich wynikach mikologicznych (2880 prób) pracowni bakteriologii spzg i chp w olsztynie. grzyby pochodziły z różnych odcinków ontocenozy układu oddechowego, pokarmowego oraz ze skóry pacjentów onkolo gicznych i z przewlekłymi chorobami układu oddechowego, w tym podejrzanych o gruźlicę. ogólnie uzyskano 317 izolatów s. cerevisiae, co stanowi 11,04% wszystkich stwierdzonych grzybów gatunek ten najczęściej towarzyszył licznym grzybom drożdżopodobnym (candida, trichosporon). ontocenozą najczęściej zasiedlaną przez s. cerevisiae był układ pokarmowy, a następnie oddechowy. na skórze notowano go głównie w ostatnich dwóch latach badań. o ile w układzie oddechowym i pokarmowym drożdże pojawiały się z podobną częstością u kobiet i mężczyzn (z niewielkim przesunięciem w kierunku kobiet), to na skórze kobiet notowano je prawie dwukrotnie częściej. z reguły izolowano je latem i jesienią, rzadko wiosną i zimą. we wszystkich ontocenozach zauważono wyraźny wzrost prewalencji badanego gatunku, co może świadczyć o wzroście jego ekspansywności w stosunku do organizmu człowieka. 2014-01-01t11:43:57+0100 polish botanical society 2014-08-20t22:53:35+0200 polish botanical society 2014-08-25t21:39:21+0200 polish botanical society 2014-08-29t18:50:04+0200 polish botanical society 2014-08-28t17:49:16+0200 polish botanical society 2014-08-29t18:51:27+0200 polish botanical society 2014-08-29t18:51:06+0200 polish botanical society 2014-08-31t21:53:40+0200 polish botanical society 2014-08-27t16:49:31+0200 polish botanical society some lichens from the vicinity of ribeiro frio (madeira, macaronesia) ivan pišút institute of botany, slovak academy of sciences, dúbravská cesta 14 sk845 23 bratislava, ivan.pisut@savba.sk pišút i.: some lichens from the vicinity of ribeiro frio (madeira, macaronesia). acta mycol. 44 (2): 179–184, 2009. a list of 49 lichens recently collected in the vicinity of ribeiro frio in madeira is given. the species ramalina nodosa and peltigera neckeri are probably new to the island. collections of cladonia stereoclada, hypogymnia madeirensis, megalospora maderensis, peltigera degenii and phlyctis agelaea are interesting as well. key words: lichenized fungi, madeira, portugal introduction madeira is a portuguese island in the atlantic ocean, 900 km sw of portugal and 660 km nnw of the canary islands (fig. 1). in winter 2002 and 2003 i had an opportunity to study lichens at some localities during my short holidays in madeira (pišút 2004). in 2008 and 2009 i visited the island again and studied the lichens of the laurel forests near the habitation of ribeiro frio situated below poiso, a mountain transition in the northern part of the island on the road between funchal and santana. in 2008 i studied the lichens along the levada between ribeiro frio and portello (1), in 2009 on the levada between ribeiro frio and balcões (2) and in the laurel forest above the habitation (3). the material is preserved in my herbarium and in lichen collections of the institute of botany, slovak academy of sciences, bratislava (sav). acta mycologica vol. 44 (2): 179–184 2009 dedicated with pleasure to professor krystyna czyżewska not only for her significant contribution to the knowledge and conservation of polish lichens, but also for her activities in cooperation networks of central european lichenologists 180 i. pišút results and discussion data on 49 lichen species are presented. the species ramalina nodosa and peltigera neckeri are probably new to the island; other findings, e.g., cladonia stereoclada, hypogymnia madeirensis and megalospora maderensis, are also interesting. byssoloma leucoblepharum (nyl.) vain. − on leaves of laurus azorica above ribeiro frio, alt. ca 920 m (3). b. subdiscordans (nyl.) p. jameson − on leaves of laurus azorica 2 km east of ribeiro frio, alt. ca 830 m (1); on leaves of laurus azorica and ilex canariensis above ribeiro frio, alt. ca 900 m (3). cetrelia olivetorum (nyl.) w. l. culb. & ch. f. culb. − on tree bark (lauraceae) ca 1 km east of ribeiro frio, alt. 850 m (1). cladonia rangiformis hoffm. − among mosses on basaltic rocks, ca 2 km eastwards, alt. 830 m, and near ribeiro frio, alt. 880 m, 2008 (1); c. stereoclada des abb. − on mossy basaltic rocks ca l km, alt. ca 850 m, and on earth ca 2 km, alt. ca 830 m, east of ribeiro frio (1); on mossy basaltic rocks above ribeiro frio, alt. ca 900 and 920 m (3), sparse. this macaronesian endemic species was described by des abbayes (1946) from the faial island in the archipelago of the azores. he characterized it as follows: “cette intéressante espèce est incontestament voisine de cl. furcata, notamment par ses réactions et ses conidanges. mais elle en est distincte par ses podétions beacoup plus ténus, semi-pellucides, et surtout par sa couche chondroïde formant un axe plein, et lorsque´elle est creuse, toujours très épaisse par suite de l´exigüité de la cavité centrale”. c. stereoclada was recorded in madeira by tavares (1952) for the first time (4 localities on mossy earth at alt. 828−1000 m) and in the canaries by østhagen and krog (1976). fig. 1. sketch map of madeira. areas above 1000 m altitude within the internal line. adapted from arvidsson and wall (1985). some lichens from ribeiro 181 degelia plumbea (lightf.) p. m. jørg. & p. james − on humid basaltic rocks and on tree bark (lauraceae) 2 km east of ribeiro frio, alt. 830 m (1). fellhanera bouteillei (desm.) vězda − on leaves of laurus azorica 2 km eastwards, alt. ca 830 m (1), and above ribeiro frio, alt. ca 920 m (3). heterodermia leucomelos (l.) poelt − on branches and the bark of various laurels and on mossy basaltic rocks, alt. 850-880 m (1), on the bark of various lauraceae, alt. 900 m; abundant (3). hypogymnia madeirensis (tav.) hawks. − on dry branches of a deciduous tree (lauraceae) ca 2 km east of ribeiro frio, alt. ca 830 m (1). my specimen lacks soredia, contains a few stalked apothecia and its medulla is pd−. h. madeirensis shows some superficial similarity to h. tavaresii, but the latter is larger and its medulla reacts pd+ bright yellow (hawksworth 1973). tavares (1952) recorded this species (as parmelia madeirensis) from its locus classicus (“inter queimadas et calderario verde, ad ramos ericae circ. 900 m s. m. 9−viii−1951”). hypotrachyna endochlora (leight.) hale − on ocotea foetens bark ca 1 km east of ribeiro frio, alt. ca 850 m (1). from madeira for the first time recorded by arvidsson and wall (1985) in ribeiro frio; h. laevigata (sm.) hale on mossy basaltic rocks ca 2 km east of ribeiro frio, alt ca 830 m (1). from madeira for the first time recorded as parmelia laevigata by alstrup (1991) in ribeiro frio; h. pulvinata (fée) hale − among mosses on basaltic rocks 2 km, alt. ca 830 m and 1 km, alt. ca 850 m, east of ribeiro frio (1). probably already recorded by tavares (1964, as h. taylorensis). h. pulvinata is absent in madeira in hafellner´s checklist (1995), but it occurs there according to schumm (2008). the only difference between h. pulvinata and h. taylorensis is the upper surface of older specimens of the latter with occasional irregular white abraded areas. they are probably symptoms of altering according to schumm (l.c.). remark. if both species are identical, the binom parmelia pulvinata fée 1824 has priority against parmelia taylorensis m. e. mitsch. 1961; h. rockii (zahlbr.) hale − on the bark of laurus azorica, 2 km east of ribeiro frio, alt. ca 830 m (1). for the first time recorded from madeira in the laurel forest (several collections) by arvidsson and wall (1985). leptogium brebissonii mont. − on mossy basaltic rock above ribeiro frio, alt. ca 900 m (3); l. cochleatum (dick.) p. m. jørg. & p. james − on mossy basaltic rocks and on the bark of laurus azorica 2 km east of ribeiro frio, alt. 830 m (1); on cedar bark near ribeiro frio, alt. 880 m (1); on mossy basalt rock above ribeiro frio, alt. ca 900 m (3); l. cyanescens (rabh.) körb. on cedar bark near ribeiro frio, alt. ca 880 m (1). lobaria immixta vain. − on branches of ocotea foetens 2 km east of ribeiro frio alt. ca 830 m (1); on branches of ocotea foetens and laurus azorica above ribeiro frio, alt. ca 900 m (3); l. sublaevis (nyl.) tav. − on the bark of a deciduous tree (lauraceae) 2 km east of ribeiro frio, alt. 830 m (1); on fallen tree branches (lauraceae) between ribeiro frio and balcões, alt. 850 m (2); on the bark of laurus azorica above ribeiro frio, alt. ca 900 m (3). megalospora maderensis (krempelh.) sipman − on the bark of ocotea foetens ca 1 km east of ribeiro frio, alt. ca 850 m (1). this rare endemic species in madeira is characterized with by bacillar spores, the presence of clearly thickened edges around the spore septa and the presence of 182 i. pišút usnic acid (sipman 1983). it occurs at elevations from 850 to 1000 m on bark and rock (tavares 1952, as bombyliospora amplificans var. maderensis). nephroma foliolatum p. james & f. j. white − on fallen branches of a deciduous tree (lauraceae), alt. ca 850 m (2); among mosses on the bark of laurus azorica above ribeiro frio, alt. ca 900 m (3). according to james and white (1987), nephroma foliolatum is confined to the laurel forest of madeira. it is also known from a single unlocalized gathering collection from the canary islands; n. laevigatum ach. − on a wooden fence near ribeiro frio, alt. ca 880 m (3). normandina pulchella (borrer) nyl. − on the thallus of degelia plumbea growing on tree bark (lauraceae) above ribeiro frio, alt. ca 900 m (3); as a facultative lichen growing on lichens. ochrolechia pallescens (l.) a. massal. − on fallen branches of deciduous trees (lauraceae) ca 2 km east of ribeiro frio, alt. 830 m, and near ribeiro frio, alt. 880 m (1). pannaria rubiginosa (ach.) bory − among mosses on the bark of laurus azorica, alt. ca 850 m (2); on the bark of ocotea foetens above ribeiro frio, alt. ca 900 m (3). parmelia saxatilis (l.) ach. − on mossy basaltic rock above ribeiro frio, alt. ca 900 m (3). parmotrema arnoldii (du rietz) hale − on tree bark (lauraceae), ca 2 km east of ribeiro frio, alt. 830 m (1); p. perlatum (huds.) m. choisy − on branches of a deciduous tree (lauraceae) near ribeiro frio, alt. 880 m, (1); on mossy basaltic rock above ribeiro frio, alt. ca 900 m (3); p. robustum (degel.) hale − on tree bark (lauraceae), ca 1 km east of ribeiro frio, alt. ca 850 m (1). peltigera degenii gyeln. − among mosses on basaltic rock 1 km east of ribeiro frio, alt. 850 m (1). in madeira so far recorded only on mossy stones along the road in cruzinhas, at alt. ca 550 m (alstrup 1991); p. membranacea (ach.) nyl. − on mosses on basaltic rock above ribeiro frio, alt. ca 900 m (3); p. neckeri hepp ex müll. arg. − on mossy basaltic rocks near ribeiro frio, alt. 880 m (1). pertusaria leucostoma (bernh.) a. massal. − on fallen branches of a deciduous tree (lauraceae), ca 1 km east of ribeiro frio, alt. ca 850 m (1); p. ophthalmiza (nyl.) nyl. − on branches of laurus azorica, 2 km east of ribeiro frio, alt. ca 830 m (1); on the bark of ocotea foetens between ribeiro frio and balcões, alt. ca 850 m (2). from the second site probably already recorded by tavares (1952) on introduced platanus (as p. multipuncta). phlyctis agelaea (ach.) flot. − on dry braches of ocotea foetens ca 2 km east from ribeiro frio, alt. ca 830 m (1). for the first time recorded on flakes of eucalyptus between camacha and carreiras by arvidsson and wall (1985). porina leptosperma müll. arg. − on leaves (laurus azorica), alt. ca 920 m (3). according to sérusiaux (1996), this otherwise pantropical species is common on the island. pseudocyphellaria aurata (ach.) vain. − on the bark of a deciduous tree (lauraceae) 2 km east of ribeiro frio, alt. 830 m (1); on the bark of ocotea foetens above ribeiro frio, alt. ca 900 m (3); p. crocata (l.) vain. − on dry branches of a deciduous shrub above ribeiro frio, alt. ca 900 m (3); p. intricata (delise) vain. − on the bark of laurus azorica above ribeiro frio, alt. ca 900 m (3). some lichens from ribeiro 183 ramalina farinacea (l.) ach. − on fallen tree branches (lauraceae), alt. ca 900 m (3); r. nodosa krog & østhagen − on almost vertical basaltic slopes ca 2 km east of ribeiro frio, alt. ca 830 m (1). probably new to madeira, according to krog and østhagen (1980) and aptroot and schumm (2008); until now known only from the canary islands (fuerteventura, tenerife and gomera). ramalina nodosa is characterised by delicate, thin (0.1−1mm), richly branched subterete to terete laciniae without soredia and pseudocyphellae, with numerous nodules; r. subgeniculata nyl. − on fallen tree branches (lauraceae), 1 km east of ribeiro frio, alt. 850 m (1); in the levada between ribeiro frio and balcões, alt. 850 m (2) and above ribeiro frio, alt. ca 900 m (3). rimelia reticulata (taylor) hale & a. fletscher − on a deciduous tree (lauraceae) ca 1 km east of ribeiro frio alt. 850 m (1); on the bark of ocotea foetens between ribeiro frio and balcões alt. ca 850 m (2). stereocaulon azoreum (schaer.) nyl. − on basaltic rocks l km eastwards, alt. 850 m (1) and above ribeiro frio, alt. ca 900 m (3). sticta canariensis bory ex del. − on tree branches (lauraceae) l km east of ribeiro frio, alt. 850 m (l) and abundantly on mossy rocks above ribeiro frio, alt. ca 900 m (3). tapellaria epiphylla (müll. arg.) r. sant. − very frequent, e.g., on leaves of laurus azorica 2 km east of ribeiro frio, alt. ca 830 m (1); on leaves of ocotea foetens, ilex canariensis, ilex perado, persea indica and introduced taxus baccata above ribeiro frio, alt. ca 900 m (3). thelotrema lepadinum (ach.) ach. − on the bark of ocotea foetens between ribeiro frio and balcões, alt. ca 850 m (2). usnea articulata hoffm. − on fallen tree branches (lauraceae) above ribeiro frio, alt. 900 m (3). usnea rubicunda stirton − on fallen tree branches (lauraceae) 2 km, 830 m and l km east of ribeiro frio, alt. ca 850 m (1); on fallen tree branches (lauraceae) above ribeiro frio, alt. 900 m (3). woessia apiahica (müll. arg.) sérus. − on tree leaves (lauraceae) above ribeiro frio, alt. ca 900 m (3). in madeira already mentioned by santesson (1952) as bacidia apiahica. acknowledgement. i thank my son ing. p. pišút, phd for his help in the field and for his comments on the manuscript. 184 i. pišút references abbayes, des h. 1946. les cladonia (lichens) des iles açorés. port. acta biologica 1 (3/4): 243–254. alstrup v. 1991. some lichens from madeira. graphis scripta 3 (3): 108–109. aptroot a. f., schumm f. 2008. key to ramalina species known from atlantic islands, with two new species from the azores [bestimmungsschlüssel für ramalina arten von den atlantischen inseln, mit zwei neuen arten aus den azoren]. sauteria 15: 21–57. arvidsson l., wall s. 1985. contribution to the lichen flora of madeira. lichenologist 17: 39–49. hafellner j. 1995. a new checklist of lichens and lichenicolous fungi of insular laurimacaronesia including a lichenological bibliography for the area. fritschiana 5: 1–132. hawksworth d. l. 1973. two new species of hypogymnia (nyl.) nyl. lichenologist 5: 452–456. james p. w., white f. j. 1987. studies on the genus nephroma i. european and macaronesian species. lichenologist 19: 215–268. krog h., østhagen h. 1980. the genus ramalina in the canary islands. norw. j. bot. 27: 255–296. østhagen h., krog h. 1976. contribution to the lichen flora of the canary islands. norw. j. bot. 23: 221–242. pišút i. 2004. a few lichens from madeira. acta rer. natur. mus. nat. slov. 50: 14–17. santesson r. 1952. foliicolous lichens i. a revision of the obligately foliicolous, lichenized fungi. symbolae bot. upsalienses 12 (1): 1–500. schumm f. 2008. flechten madeiras, der kanaren und azoren. ed. beck, ottg süssen, 294 pp. sérusiaux e. 1996. foliicolous lichens from madeira with the description of a new genus and two new species and a world-wide key of foliicolous fellhanera. lichenologist 28 (3): 197–227. sipman h. m. 1983. a monograph of the lichen family megalosporaceae. biblioth. lichenol. 18: 1–241 and tables. tavares c. n. 1952. contributions to the lichen flora of macaronesia i – lichens from madeira. port. acta biol. sér. b, sist. 3 (3): 308–391. tavares c. n. 1964. contributions to the lichen flora of macaronesia iii – new or interesting taxa. revista de biologia 4 (1/2): 131–144. o niektórych porostach poznanych w okolicach ribeiro frio (madera, makaronezja) streszczenie podano listę 49 gatunków porostów zebranych w latach 2008 i 2009 w okolicy siedziby ribeiro frio na wyspie madera. gatunki ramalina nodosa i peltigera neckeri są prawdopodobnie nowe dla tej wyspy; interesujące są równieź notowania gatunków cladonia stereoclada, hypogymnia madeirensis, megalospora maderensis, peltigera degenii i phlyctis agelaea. 2014-01-01t11:49:21+0100 polish botanical society 2014-08-26t20:31:27+0200 polish botanical society 2014-08-28t14:43:39+0200 polish botanical society 2014-08-31t21:54:47+0200 polish botanical society 2014-08-31t21:55:36+0200 polish botanical society yeast-like fungi isolated from indoor air in school buildings and the surrounding outdoor air elżbieta ejdys, joanna michalak and katarzyna monika szewczyk department of mycology, university of warmia and mazury in olsztyn oczapowskiego 1a, pl-10-719 olsztyn-kortowo, elzbieta.ejdys@uwm.edu.pl ejdys e., michalak j., szewczyk k. m.: yeast-like fungi isolated from indoor air in school buildings and the surrounding outdoor air. acta mycol. 44 (1): 97–107, 2009. a total of 111 isolates of yeast-like fungi and yeasts belonging to 40 species of 19 genera were identified in indoor air and outdoor air. only one species, kluyveromyces marxianus, was recorded in both types of air and seasons (spring and autumn). kluyveromyces lactis and yarrowia lipolytica, a species having the greatest symbiotic abilities, dominated in indoor air and outdoor air, respectively. intensely used rooms, especially those with limited access of air, have the broadest range of species of yeast-like fungi. a comparison of both habitats shows that school rooms pose a greater epidemiological risk of yeast-like infections than outdoor air. the indoor as well as outdoor mycobiota undergoes phenological changes although it is determined by other biotic and abiotic factors. key words: yeast-like fungi, school, indoor air introduction fungal spores and mycelium fragments constitute the greatest part of bioaerosols in indoor air (krzysztofik 1992). they use air as a vector between living organisms and water, soil and abiotic ecosystem components (dynowska, ejdys 1999). similarly to outdoor air, indoor air aerosol contains biological particles. fungal spores in indoor air are consistent with the seasonal development of individual species in nature (twarużek 2006). the accumulation of cellulose and lignin sources: construction wood, furniture (zyska 2001), plywood, paper, fireboard and chipboard (miklaszewska, grajewski 2006), as well as other sources: plaster, paint (zyska 2001; grajewski, twarużek 2004), provides favourable conditions for the development of indoor fungi. although the taxonomical range of the mycobiota of different types of air is very broad, moulds dominate in it (zyska 2001; ejdys 2007; topbas et al. 2006). however, the acta mycologica vol. 44 (1): 97–107 2009 98 e. ejdys et al. prevalence of yeast-like fungi does not exceed 10% in indoor air in buildings having normal humidity and temperature parameters (ejdys 2007) and can reach a maximum of 20% in particularly damp rooms (meklin et al. 2002). this is surprising given special enzymatic abilities and a high rate of proliferation and growth of yeastlike fungi. they have been isolated from biotic and abiotic ecosystem components in all climatic zones (dynowska 1995; kurnatowska 1998; bogacka, matkowski 2001; arias et al. 2002; nagornaya et al. 2003; biegańska 2006). a large group of potentially pathogenic species mostly of the genera candida, kluyveromyces, debaryomyces, rhodotorula, pichia, trichosporon, sacharomycopsis (dynowska 1995) and recently also saccharomyces (dynowska et al. 2006) or zygoascus (brandt et al. 2004) has been identified within yeast-like fungi. they are usually an aetiological factor in mycoses and, less frequently, in mycoallergies. as they can produce toxins (e.g. candotoxins), the species can cause mycotoxycoses and increase micro-organism sensitivity to some bacterial infections (dynowska 1995). age is the main physiological factor of the school period that predisposes to fungal infections. the risk of mycoses increases as other epidemiological factors occur, e.g. a child spends time in a large same-age group in the same rooms for several hours. children spend between 5 and 9 hours on the school premises daily and it is of mycological importance how clean school buildings and their surroundings are. heavier than mould spores, spores of yeast-like fungi are located in the lower part of the air column (stach et al. 2004). they can rise up to a height of >1.5m as a result of air currents related to human movement (kurnatowska 1999). the two most important infection portals, the nose and the mouth, are located at this height in primary school children. therefore, a high accumulation of yeast-like fungi in the air can pose a serious threat to the health of children inhaling this air (kurnatowska 1999; meklin et al. 2002). material and methods schools in olsztyn (165 000 inhabitants) and dobre miasto (11 000 inhabitants) were selected. both olszytn and dobre miasto are situated in the pojerzezie olsztyńskie lakeland. lower temperatures and a shorter vegetation period in comparison with other parts of poland are observed in the area (kondracki 2002). yeast-like fungi occurring temporarily in outdoor and indoor air were examined. fungi were collected in may (central heating switched off) and in november (heating operating). koch sedimentation method was used to obtain the material. petri dishes with both types of sabouraud’s medium (chloramphenicol and rose-bengal) were exposed at each site in three replicates. thirteen sites were established around the school building and the gymnasium in olsztyn (school a) and twelve in dobre miasto (school b). dishes were placed at building corners, at half the wall length, 1.5 m away from the wall. samples were also taken from the internal patio in school a. the exposure time was 10 min. a total of 13 different areas inside the school were selected on the ground floor (classroom, women’s toilet, men’s toilet, corridor and yeast-like fungi 99 sports changing room) and in the basement (cloakroom) as well as in the newly refurbished common room for pe teachers (school a only). samples containing both media were placed directly on the floor diagonally in two corners of each room. the exposure time was 30 min. humidity and temperature were measured at each site. samples were incubated at 37°c for the first three days and transferred to room temperature for 72 hours. isolates of yeast-like fungi and yeasts were inoculated on sabouraud’s medium without antibiotics. media containing antibiotics that could affect typical fungal properties were not used (dynowska 1991-1992). after pure bacteria-free strains were obtained, microcultures were established on nickerson agar. fungi were inoculated on glass slides covered with a thin layer of the medium (ca. 2 mm). two-three drops of broth with serum (1:1) were placed on the inoculation site. microcultures were incubated at 37°c for a period from 48 to 72 hours. macroscopic features (size, colour, shape, texture, smell of the colony), microscopic features (shape and size of budding cells, blastospores and chlamydospores, the diameter of pseudohyphae and hyphae) and biochemical features obtained on bio–mérieux api-tests (api 20c, api 20c aux) were used for identification. chromagar (bio–mérieux) was used to differentiate individual species of the genus candida. chromagar was treated as an auxiliary test as it does not yield accurate results in mixed-species isolations (bouchara et al. 1996). photographic documentation was made throughout the process. yeast-like fungi and yeasts were determined using the following keys: kreger van rij (1984), barnett, payne and yarrow (1990), and kurtzmann and fell (2000). results a total of 111 isolates of yeast-like fungi and yeasts belonging to 40 species (+ 1 undetermined) of 19 genera were obtained (tab. 1). the greatest numbers of representatives were recorded for the genera kluyveromyces and pichia (6 species), followed by candida (5), debaryomyces (4) and rhodosporidium (2). other genera occurred incidentally. kluyveromyces was one of the most frequently recorded genera (24 isolates) as regards the quantity; fungi of the genera debaryomyces and pichia were recorded slightly less frequently (18 and 15 isolates, respectively). kluyveromyces lactis dominated in both schools. it was, however, isolated only from indoor air when central heating was on. d. polymorphus was also recorded only in autumn, but it was present indoors and outdoors. only one species, kluyveromyces marxianus, was recorded in both types of the air and in both seasons. three groups of survival in air were distinguished: i – species isolated only from indoor air, ii – species isolated only from outdoor air, and iii (tab. 1) – species that survived in both types of air aerosol. group i was the most numerous one (20 of the 40 taxa identified in the study). group iii was only 20% and comprised the following fungi: aureobasidium pullulans, debaryomyces occidentalis, d. polymorphus, kluyveromyces marxianus, oosporidium margaritiferum, pichia membranifaciens, saccharomycopsis capsularis, yarrowia lipolytica. although it was recorded six times 100 e. ejdys et al. table 1 yeast-like fungi in may and november in indoor air and outdoor air no species number of isolates indor air outdoor air school a school b around a around b may* nov. may nov. may nov. may nov. 1 arxula adeninivarans van der walt, smith & yamada 1iii** 2 a. terrestris van der wald, smith &yamada 1 3 aureobasidium pullulans arn 2ii,v 2 3 4 candida albicans berkhout 1 5 c. glabrata meyer & yarrow 1v 1ii 6 c. rhagii jurzitza, kühlwein & kreger-van rij 1ii 7 c. solani lodder & kreger-van rij 1iii 8 candida utilis lodder & kreger-van rij 1ii 9 cystofilobasidium informominiatum hamamoto, sugiyama & komagata 3iv,v 1vii 10 debaryomyces hansenii lodder & kregervan rij 1v 1i 3ii,v 11 d. occidentalis kurtzman & robnett 1iv 1 12 d.polymorphus price & phaff 1ii 4i,iii,vi 2 3 13 d. venrijiae abadie, pignal & jacob 1ii 14 geotrichum clavatum de hoog, smith & gueho 2 15 kluyveromyces blattae henninger & windisch 1 16 k. lactis van der walt 6iii,iv,v,vi 6ii,iii,iv,v 17 k. marxianus van der walt 3i,iii 1 1 1 2 18 k. thermotolerans yarrow 1i 1iii 19 k. wickerhamii van der walt 1v 1v 20 k. yarrowii van der walt, johannsen, opperman & halland 1ii 21 kluyveromyces sp. 1 22 leucosporidium scottii fell, statzell, hunter & phaff 1 23 lipomyces starkeyi lodder & kreger-van rij 1v 24 mrakia frigida yamada & komagata 1iv 1ii 25 nadsonia commutata golubev 2v 26 oosporidium margaritiferum stautz 1vi 1 27 pichia anomala kurtzman 1ii 28 p.burtonii boidin, pignal, lehodey, vey & abadie 1 29 p.delftensis bebech 1 30 p.farinosa hansen 2iv,v 2iv,vi 3ii,v,vi 31 p.guilliermondii wickerham 2 32 p.membranifaciens hansen 1vi 1i 1 33 rhodosporidium dacryoideum fell, hunter & tallman 1iii 34 r. toruloides banno 1iv 35 rhodotorula glutinis harrison 1v 36 saccharomyces bayanus saccardo 1 37 saccharomycopsis capsularis schiönning 1vi 1 38 s.vini van der walt & scott 2 39 yarrowia lipolytica van der walt & von arx 2ii,vi 6 40 zygoascus hellenicus smith 1 2 41 zygosaccharomyces rouxii yarrow 3 11 13 12 30 6 15 8 16 total 66 45 abbreviations: *isolation month; ** number of isolates indoors: i-classroom, ii-women’s toilet, iii-men’s toilet, iv-corridor, v-cloakroom, vi-sports changing room, vii-common room (pe teachers). 102 e. ejdys et al. the broadest taxonomical range of fungi in indoor air was recorded in the cloakroom (14 species) and in the women’s toilet (13 species). fungi isolated in the cloakroom did not occur in outdoor air (tab. 1). fewer species were recorded in the air in the corridor, men’s toilet and sports changing room. the abundance of fungi in the classroom was almost three times smaller (five species): d. hansenii, d. polymorphus, k. marxianus, k. thermotolerans and p. membranifaciens. only one species, cystophilobasidium informominiatum, was identified in the pe teachers’ common room. kluyveromyces lactis was recorded in the air of five different rooms. pichia farinosa was isolated slightly less frequently (4 rooms). a certain relationship between the occurrence of yeast-like fungi and the site’s microgeographic location was observed (tab. 3). the greatest specific range was observed at eastern sites. a total of 22 isolated were recorded in the east, north-east and southeast. no fungi examined in the study were recorded in the samples at northern sites. the amplitude of indoor air was the same in spring and in autumn when central heating was on (2.2°c) and ranged from 19.4°c to 21.6°c and from 18.4°c do 20.6°c, respectively. relative air humidity ranged from 48% and 60% regardless of the season. temperature and humidity of outdoor air ranged considerably depending on site location. temperature ranged from 21.2 to 33.6°c in olsztyn and from 18.2 to 22.7°c in dobre miasto in spring, and was (14.2:22.7) and (8.3:14.2) in autumn. air humidity was 33-67% (olsztyn) and 42-50% (dobre miasto) in may. it was higher in november and was 46-65% in olsztyn and 62-92% in dobre miasto. table 3 species diversity and the directions of the world north-west north north-east species school, isolation month species school, isolation month species school, isolation month d. polymorphus z. hellenicus a. pullulans s-1,nov.* s-1,may s-1,may a. terrestris d. polymorphus k. blattae s. bayanus z. hellenicus c. albicans d. polymorphus s. capsularis s. vini y. lipolytica s-1,nov. s-1,nov. s-1,nov. s-1,nov. s-1,nov. s-2,nov. s-2,nov. s-2,nov. s-2,nov. s-2,nov. west patio east k. marxianus p. guilliermondii p. membranifaciens a. pullulans d. occidentalis k. marxianus o. margaritiferum s-1,nov. s-1,nov. s-1,nov. s-2,may s-2,may s-2,may s-2,may a. pullulans g. clavatum s-2,may s-2,may p. burtonii p. guilliermondii z. rouxii a. pullulans l. scottii p. delftensis d. polymorphus k. marxianus y. lipolytica s-1,nov. s-1,nov. s-1,nov. s-1,may s-1,may s-1,may s-2,nov. s-2,nov. s-2,nov. south-west south south-east g. clavatum kluyveromyces sp. y. lipolytica s-2,may s-2,nov. s-2,nov. z. rouxii k. marxianus s-1,nov. s-1,may a. pullulans s. vini y. lipolytica s-1,may s-2,nov. abbreviations: *s-1 school 1, s-2 school 2. yeast-like fungi 103 discussion current studies usually focus on threats posed by yeast-like fungi to human and animal health. studies in applied mycology related to human economy are numerous (hahn et al. 2006; golubev et al. 2003; angerbauer et al. 2008). however, up-to-date reports on basic research into natural habitats of yeast-like fungi that concentrates on humans are scarce. it is therefore difficult to determine whether 40 species of yeast-like fungi isolated in the study in urban conditions in central europe are an exceptional figure or if they are typical of this environment. the lack of reports on the occurrence of yeast-like fungi in air samples may be connected with methodological problems. cultures should not be interfered with in the initial phase of incubation to avoid sample contamination and to identify the full mycobiota range. it is often possible to isolate yeast-like fungi only at the beginning of culture as fast-growing moulds destroy them. the use of substrates suitable for the development of moulds (czapek-dox) recommended in the polish standard (pn 89/z04111/03) additionally promotes them. incubation temperature (26°c) also promotes the majority of moulds and not only slightly more thermophilous fungi that also include some yeast-like fungi. for instance, arxula adeninivorans, which can grow up to a temperature of 48°c (wartman, kunze 2000; kunze 2006), was obtained in the study by using an incubation temperature higher (37°c) than that recommended for the examination of air mycobiota. a broad temperature range seems to be a key factor in the ecological success of kluyveromyces marxianus, which was recorded in both types of air in spring and autumn. it can survive in different temperature ranges and can therefore function in water, on plants or in human or animal bodies (dynowska 1995; kurtzmann, fell 2000). broad enzymatic abilities allow it to ferment available sugars and different sources of carbon of both plant and animal origin. its ability to colonise different ecological niches may also be attributed to the type of its reproduction. kluyveromyces marxianus combines rapid reproduction by budding in favourable conditions with alterations of genetic information during unfavourable environmental changes by producing ascospores. another ascomycete, kluyveromyces lactis, occurs mostly in milk and milk-derived products (kurzman, fell 2000). its relatively rare ability to ferment and assimilate lactose allows the species to dominate in environments where it is available. small cartons of short-lifetime milk were distributed at school and the species was prevalent in the study. food remains may also have been a source of and/or substrate for debaryomyces occidentalis, d. hansenii or mrakia frigida. although they are used in biotechnology and the food industry (mazurkiewicz-zapałowicz 2006), they can also contaminate ready products (moreira et al. 2001). aureobasidium pullulans and pichia farinose, species dominating in school b in spring, may have come from outdoors. soil, cereal crops and their products are natural environments of occurrence of these fungi (twarużek 2006; kurzman, fell 2000). pupils or the staff may also have been a source of them as these fungi have been isolated from people (salkin et al. 1986; bolignano, criseo 2003; garcia-martos et al. 1996). the presence of a. pullulans in indoor air can intensify asthma symptoms (niedorzytko et al. 2007) or lead to the development of pulmonary hypersensitivity to its spores (temprano et al. 2007). 104 e. ejdys et al. conditions favourable for the occurrence and existence of different fungi are identified in intensely used rooms with a high through-put of people (cloakroom, walkways or passages). a broad range of fungi belonging to different systematic and ecological groups vegetates on organic matter from occupants’ bodies (dead epidermis, saliva, hair) or clothes, shoes or functional items. as the occurrence of moulds having especially high toxinogenic abilities in such rooms shows, increased competition is also associated with mycobiota richness (ejdys 2008). organic compounds formed from lysis of dead fungal and bacterial cells accumulate in indoor aerosol with an abundant mycobiota. they can provide a particularly suitable substrate for the development of yeast-like fungi. fungi can increase survival potential in difficult environmental conditions by expanding their enzymatic range and entering symbiosis with other organisms, even other fungi. yarrowia lipolytica occurred exclusively with other fungi in the study. co-occurring species have a broader range of fermentation and assimilation abilities than y. lipolytica, which does not ferment and its carbon sources are limited only to glucose and partly galactose. the occurrence of organisms in the same habitat may show similar or overlapping parameters of their ecological niches. fungi such as cystofilobasidium infirmominiatum, rhodotorula glutinis and debaryomyces hansenii were isolated from air in the cloakroom in the study. this seems to be connected with the fact that these fungi prefer similar organic compounds as the same set of fungi has been isolated from poultry meat (deak 2001) and from human clinical material (kurzman, fell 2000; desnos-ollivieri et al. 2008; ejdys 2008). although the species have been isolated from plant material (kurzman, fell 2000; golubev et al. 2003), their presence in school buildings is connected with the human body and its excretions or secretions rather than with plant tissues and can pose a potential threat to the health of building occupants. relatively constant temperature and humidity conditions in relation to outdoor sites are observed indoors. moreover, light access is limited even on sunny days in the cloakrooms which are located in the basement. this may explain the indoor occurrence of yeast-like fungi that were not recorded in outdoor air. they may belong to “domestic fungi”, such as aspergillus or penicillium, especially as permanent exposure of the cloakroom to outdoor air did not considerably influence the composition of the mycobiota of yeast-like fungi as shown by the absence of species that occurred in outdoor air at the same time. yeast-like fungi that occurred only in outdoor air samples may be treated as “field fungi”. these fungi or only some of their strains are less sensitive to high daily temperature and humidity amplitudes, and, primarily, a large amount of ultraviolet radiation. they found the most favourable growth conditions at eastern localities where, while there is much light in autumn, temperature is higher than that at northern sites. some fungi preferred western sites in spring. phenological fluctuations are observed in the occurrence of fungi in outdoor air. the air is mycologically cleaner in winter than in summer. a greater number of spores are recorded in air in spring and autumn than in winter and summer (kruczalak et al. 2002). however, the greatest number of fungi in air is observed in autumn when all spring and autumn months are examined (shelton et al. 2002; daccaro et al. 2003). the results confirm this also in indoor air. there were fewer yeast-like yeast-like fungi 105 fungi indoors in spring than in autumn and the species were not the same as those when the heating was on. a decrease in building insulation by airing did not affect considerably the composition of the mycobiota as a small percentage of fungi occurred both indoors and outdoors. phenological changes of the indoor mycobiota were most probably caused not as much by changes of abiotic factors (temperature, radiation, humidity) as is the case with the external environment but by occupantrelated biotic factors. pupils and the staff adapt their clothing and nutrition to the season; their immune systems also change. changes in habitat parameters determine reorganisation of the species composition of all microorganisms, including fungi. conclusions indoor air in school buildings having normal humidity and temperature param-1. eters can abound in yeast-like fungi: as many as 28 species of yeast-like fungi belonging to 15 genera were recorded. a total of 20 species belonging to 14 genera were identified in outdoor air. kluyveromyces lactis2. dominated in indoor air while yarrowia lipolytica, a species characterised by the greatest symbiotic abilities, dominated in outdoor air. a comparison of both habitats shows that a greater epidemiological risk of infec-3. tion by yeast-like fungi is observed indoors rather than outdoors. intensely used rooms, especially where light access is limited, have the broadest 4. spectrum of species of yeast-like fungi and therefore pose the greatest potential risk to the health of children and the school staff. indoor and outdoor air mycobiota changes phenologically although it is deter-5. mined by other biotic and abiotic factors. species of yeast-like fungi that occur only indoors may be classified as “domes-6. tic fungi” while species recorded in outdoor air may be considered to be “field fungi”. references angebauer c., siebenhofer m., mittelbach m., guebitz g.m. 2008. conversion of sewage sludge into lipids by lypomyces for biodisel production. bioresource technology 99 (8): 3051–3056. arias c.d., burns j.k., friedrich l.m., goodrich r.m., parish m.e. 2002. yeast species assiociated with orange juice: evaluation of different identitication methods. appl. and environ. microbiol. 68 (4): 1955–1961. barnett j.,a., payne r., yarrow d. 1990. yeasts : characteristics and identification. cambridge univ. press. biegańska m.j. 2006. drożdżaki z rodzaju candida – nie zawsze patogenne, ale... mikol. lek. 13 (4): 318–321. bogacka e., matkowski k. 2001. wpływ grzybów na zdrowie ludzi. mikol. lek. 8 (3/4): 175–178. bouchara j.p., declerck p., cimon b., planchenault c., de gentile l., chabasse d. 1996. routine use of chrom agar candida medium for presumptive identification of candida yeast species and detection of mixed fungial populations. clin microbiol. infect. 2 (3): 202–208. brandt m.e., kauffman c.a., pappas p.g., iqbal n., arthington-skagga b.a., lee-yang w., smith m. 2004. fungemia caused by zygoascus hellenicus in an allogeneic stem cell transplant recipient. j.clin microbiol. 42 (7): 3363–3365. 106 e. ejdys et al. dacarro c., picco a.m., grisoli p., rodolfi m. 2003. determination of aerial microbiological contamination in scholastic sports environments. j. app. microbiol. 95: 904–912. deak t. 2001. identification of yeasts isolated from poutry meat. acta biol. hungar. 52 (2/3): 195–200. desnos-ollivier m, ragon m, robert v, raoux d, gantier jc, dromer f. 2008. debaryomyces hansenii (candida famata), a rare human fungal pathogen often misidentified as pichia guilliermondii (candida guilliermondii). j clin microbiol. 46 (10): 3237–3242. dynowska m. 1991-1992. the influence of antibiotics on the morphology of candida albicans and candida stellatoidea. acta mycol. 27 (1): 205–211. dynowska m. 1995. drożdże i grzyby drożdżopodobne jako czynniki patogenne oraz bioindykatory ekosystemów wodnych. rozpr. hab. studia i materiały wsp, olsztyn: 77: 1–83. dynowska m., ejdys e. 1999. grzyby chorobotwórcze dla człowieka. biologia w szkole 1: 21–26. dynowska m., rosłan m., góralska k. 2006. sacharomyces cerevisiae in respiratory system, digestive system and on the skin in humans. acta mycol. 41(1): 139–144. ejdys e. 2007. fungi isolated in school buildings. acta mycol. 42 (2): 245–254. ejdys e. 2008. zasady biobezpieczeństwa i higieny pracy w laboratorium badawczym a przeżywalność grzybów potencjalnie chorobotwórczych. 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(in:) j. grajewski (ed.). mikotoksyny i grzyby pleśniowe, zagrożenia dla człowieka i zwierząt. bydgoszcz: 17–34. wartman t, kunze g. 2000. genetic transformation and biotrchnological application of the yeast arxula adeninivorans. appl. microbiol. biotechnol. 54 (5): 619–624. zyska b. 2001. grzyby powietrza wewnętrznego w krajach europejskich. mikol. lek. 8 (3/4): 127–140. grzyby drożdżopodobne izolowane z powietrza budynków szkolnych i powietrza atmosferycznego wokół nich streszczenie podobnie jak w powietrzu wewnętrznym, największą część bioaerozolu powietrza atmosferycznego stanowią elementy grzybni i zarodniki grzybów z różnych grup systematycznych. duże zagęszczenie grzybów drożdżopodobnych w powietrzu może stanowić poważne zagrożenie dla zdrowia oddychająch nim dzieci. dlatego też postanowiono zbadać występowanie omawianych grzybów w powietrzu budynków szkolnych i wokół nich. materiałem do badań były grzyby drożdżopodobne występujące przejściowo w powietrzu wokół budynków szkolnych oraz w ich wnętrzu. grzyby pobierano w maju (ogrzewanie wyłączone) i listopadzie (okres grzewczy). materiał uzyskano metodą sedymentacyjną kocha, eksponując płytki petriego z podłożem sabourauda z chloramfenikolem oraz z różem bengalskim, w trzykrotnym powtórzeniu na każdym stanowisku. ogółem uzyskano 111 izolatów grzybów drożdżopodobnych i drożdży właściwych, należących do 40 gatunków zgrupowanych w 19 rodzajach. w obu porach roku i typach badanego powietrza notowano tylko jeden gatunek –kluyveromyces marxianus. stwierdzono, że powietrze budynków szkolnych, o prawidłowych parametrach wilgotności i temperatury, może obfitować w grzyby drożdżopodobne – 28 gatunków drożdżaków z 15 rodzajów. w powietrzu atmosferycznym zidentyfikowano 20 gatunków z 14 rodzajów. w powietrzu wewnętrznym dominantem był kluyveromyces lactis, natomiast w powietrzu zewnętrznym yarrowia lipolytica, gatunek o największych zdolnościach symbiotycznych. porównując badane siedliska zauważono, że w sensie epidemiologicznym, pomieszczenia szkolne stwarzają większe możliwości zakażeń grzybami drożdżopodonymi niż powietrze na zewnątrz budynków. pomieszczenia intensywnie użytkowane, zwłaszcza z limitowanym dostępem światła, mają najszersze spektrum gatunkowe drożdżaków, a tym samym stanowią największe potencjalne zagrożenie dla zdrowia dzieci i pracowników szkoły. mikobiota powietrza wewnątrz pomieszczeń i na zewnątrz ich podlega zmianom fenologicznym, chociaż jest determinowana przez inne czynniki biotyczne i abiotyczne. do „grzybów domowych” można zaliczyć gatunki drożdżaków występujące tylko w pomieszczeniach, natomiast gatunki pochodzące z powierza atmosferycznego można uznać za „grzyby polowe”. 2014-01-01t11:48:56+0100 polish botanical society 2014-08-26t20:50:01+0200 polish botanical society 2014-08-28t14:44:06+0200 polish botanical society geophilic dermatophytes and other keratinophilic fungi in the nests of wetland birds teresa korniłłowicz-kowalska1, ignacy kitowski2 and helena iglik1 1department of environmental microbiology, mycological laboratory university of life sciences in lublin leszczyńskiego 7, pl-20-069 lublin, teresa.kornilowicz@up.lublin.pl 2department of zoology, university of life sciences in lublin, akademicka 13 pl-20-950 lublin, ignacyk@autograf.pl korniłłowicz-kowalska t., kitowski i., iglik h.: geophilic dermatophytes and other keratinophilic fungi in the nests of wetland birds. acta mycol. 46 (1): 83–107, 2011. the frequency and species diversity of keratinophilic fungi in 38 nests of nine species of wetland birds were examined. nine species of geophilic dermatophytes and 13 chrysosporium species were recorded. ch. keratinophilum, which together with its teleomorph (aphanoascus fulvescens) represented 53% of the keratinolytic mycobiota of the nests, was the most frequently observed species. chrysosporium tropicum, trichophyton terrestre and microsporum gypseum populations were less widespread. the distribution of individual populations was not uniform and depended on physical and chemical properties of the nests (humidity, ph). key words: ascomycota, mitosporic fungi, chrysosporium, occurrence, distribution introduction geophilic dermatophytes and species representing the chrysosporium group (an arbitrary term) related to them are ecologically classified as keratinophilic fungi. keratinophilic fungi colonise keratin matter (feathers, hair, etc., animal remains) in the soil, on soil surface and in other natural environments. they are keratinolytic fungi physiologically specialised in decomposing native keratin. they fully solubilise native keratin (chicken feathers) used as the only source of carbon and energy in liquid cultures after 70 to 126 days of growth (20°c) (korniłłowicz-kowalska 1997). fungi other than dermatophytes and chrysosporium decompose only 30%-33% of native feather keratin in the same period (korniłłowicz-kowalska l.c.). according to kunert (2000), fungi are weakly keratinolytic if they decompose no more than 40% of keratin in liquid cultures after eight weeks and non-keratinolytic if they decompose acta mycologica vol. 46 (1): 83–107 2011 84 t. korniłłowicz-kowalska et al. less than 20%. native keratin substrates contain both keratin as well as simpler compounds, e.g., non-keratin proteins, amino acids, urea, which constitute up to 10% of the substrate’s dry weight (mercer 1958). this allows a range of other fungi that are de facto non-keratinolytic to grow on native keratin (korniłłowicz 1992). geophilic dermatophytes are represented by the genera trichophyton malmsten and microsporum gruby (anamorphs) and by their respective teleomorphs: arthroderma berk. and nannizzia stocklade. the chrysosporium group comprises two keratinolytic genera: chrysosporium corda and myceliophthora cost (anamorphs). their teleomorphs are classified in the genera arthroderma berk., aphanoascus zukal and ctenomyces eidam. or remain unknown (van oorschot 1980; currah 1985). representatives of both groups of fungi in the holomorphic stage are included in onygenales (ascomycota) and in the anamorphic stage in hyphomycetales (mitosporic fungi) (currah 1985; korniłłowicz-kowalska, wojdyło-kotwica 2008). some species of ubiquistic moulds (polyphages), and especially aspergillus fumigatus and scopulariopsis brevicaulis, also show keratinolytic abilities according to some authors (santos et al. 1996; filipello-marchisio et al. 2000) keratinophilic fungi are potentially pathogenic saprotrophs described as opportunistic pathogens (rippon 1982). pathogenic strains of these fungi, in particular species such as microsporum gypseum, m. cookei, chrysosporium keratynophilum, cause dermatomycoses in humans and animals. ubiquistic moulds with keratinolytic abilities are causal agents of opportunistic mycoses such as systemic mycoses (a. fumigatus) or superficial mycoses, e.g., nail mycoses (s. brevicaulis) (dvořak, otčenašek 1969). the biodegradation of native keratin, a protein resistant to the attack of ordinary proteolytic enzymes, works as the enzymatic lysis combined with a mechanical destruction aided by the eroding mycelium complex (english 1963, 1965; kunert 1989, 2000). in dermatophytes, it consists of the so-called frond mycelium, which erodes the substrate surface, and multicellular perforating organs penetrating the substrate and secreting keratinolytic enzymes (english 1963, 1965; kunert 2000). keratinolytic chrysosporium species produce simpler penetrative structures that are single, apically swollen hyphae known as boring hyphae (english 1963, 1965). ch. keratinophilum, which produces penetrative hyphae resembling multicellular organs in dermatophytes, is an exception (english 1969). keratinolytic chrysosporium species usually decompose native keratin more slowly than geophilic dermatophytes (korniłłowicz-kowalska 1997; filipello-marchisio 2000). ubiquistic moulds produce only thin and simple boring hyphae when growing on native keratin substrates (english 1965). the process of fungal keratinolysis consists of three stages: deamination, sulphitolysis and proteolysis (kunert 2000). deamination leads to the release of ammonia conditioned by a high nitrogen level in native keratin: from 14.72% in feathers (korniłłowicz-kowalska 1997) to 16% in hair (kunert 2000), and a narrow c:n ratio in these substrates, such as 3:1 for feathers (korniłłowicz-kowalska l.c.). n-nh4 + accumulation causes environment alkalisation necessary for enzymatic disruption of greatly numerous keratin disulphide bridges responsible for its resistance to the activity of proteolytic enzymes. sulphitolysis, that is the process of the disruption of s-s bonds, occurs with the participation of inorganic sulphite produced by the fungus (kunert 1973, 1976). this leads to keratin denaturation and, consequently, geophilic dermatophytes 85 makes proteolysis with alkaline or neutral proteases of these fungi possible (kunert 2000). during saprotrophic growth on native keratin, keratinolytic fungi oxidize 70% of carbon to co2, release 70-80% of nitrogen as ammonia and transform 30-50% of sulphur into sulphates (korniłłowicz-kowalska 1997). this allows keratinolytic fungi to play an important role in the recycling of carbon, nitrogen, sulphur of animal remains containing keratin. the occurrence of keratinophilic fungi in natural environments is conditioned primarily by their “animialisation” related to an inflow of keratin matter (montovani et al. 1982). keratin remnants are not only a nutritive source for the fungi but also a specific habitat enabling their survival and defence from other competitive microorganisms (garetta, piontelli 1975). the species diversity of keratinomycetes also depends on various physical and chemical properties of the environment, mostly ph, humidity and temperature (bőhme, ziegler 1969; chmel et al. 1972; chmel, vláčiliková 1975; garg et al. 1985; kushwaha 2000; korniłłowicz-kowalska, bohacz 2002). birds’ nests which usually contain considerable amounts of keratin matter (feathers, hair, pellets, prey remains) and have different levels of humidity and ph are therefore interesting microhabitats in this regard (pugh, evans 1970; hubalek 1974, 2000; korniłłowicz-kowalska, kitowski 2009). nests of terrestrial birds, in particular passerine passeriformes, were mostly examined in studies (conducted chiefly between 1960 and 1980) on the occurrence and distribution of geophilic dermatophytes and species of chrysosporium group related to them (pugh 1966; otčenašek et al. 1967; pugh, evans 1970; hubalek et al. 1973; hubalek 1974, 1976; hubalek, balat 1974; tokatori, hasegawa 1981). investigations on the keratinolytic mycobiota of nests of birds associated with aquatic habitats are fragmentary (hubalek 1974; korniłłowicz-kowalska, kitowski 2009). as the participation of wetland birds in the distribution of various pathogenic micro-organisms, including opportunistic pathogens causing mycoses in birds, mammals and humans has been on the increase in recent years (hubalek 2004), there is a need to expand studies on the occurrence and distribution of keratinophilic fungi in breeding and feeding biotopes of these birds. such research will contribute to a better knowledge of the role of these habitats in the survival and, partly, transmission of potentially pathogenic keratinophilic fungi. the aim of this study was to identify the species composition and the frequency of geophilic dermatophytes and chrysosporium representatives in the nests of different species of wetland birds in connection with some physical and chemical properties of those nests. material and methods nests: location, building material and structure. nests of nine bird species in southeast poland (the lublin region) were examined. a total of 38 nests were studied. the nests were collected in the period between 2006 and 2008 after they had been 86 t. korniłłowicz-kowalska et al. abandoned by birds. only nests of grey herons (ardea cinerea) were situated in the trees; the nests of other birds were in reeds and rushes of water bodies (ponds, a lake) in other aquatic vegetation or were floating nests (tab. 1). the nests were built from plant material and contained various amounts of nonplant material which was partly used to line the nest (feathers, hair) and was partly secondary (shed feathers, food remains, etc.) (tab. 2). nests of marsh harriers (circus aeruginosus) were 50-90 cm in diameter and were partly above the water surface (ca. 50 cm under water and ca. 70 cm above water). the nests were stable, non-floating, situated in reed beds of the common reed (phragmites australis) or rushes of the broadleaf cattail (typha latifolia). the base of the nests was built from twigs of black alder (alnus glutinosa), downy birch (betula pubescens) and other birches (betula sp.) as well as willows (salix sp.). nest edges were sometimes supported with stems of burdock (arctium sp.) and creeping thistle (cirsium arvense). the lining, easily distinguishable from the rest of the nest, was built from common reeds and broadleaf cattails, supplemented with sedges carex sp. and poacae grasses difficult to identify due to rotting. the lining also contained great nettle (urtica dioica), great bulrush (schoenoplectus lacustris), sometimes rhizomes of weed grass agropyron sp. and other unidentified small roots. material of animal origin constituted much of the lining: pellets of adult and young birds containing mammal hair, bird feathers, other undigested parts of the prey and prey bones; remains of uneaten prey containing hair of small mammals micromammalia, table 1 bird species, nest location and collection date no bird species systematic classification (order, family) number of nests nesting site collection place and date 1 marsh harrier circus aeruginosus l. falconiformes accipitridae 3 ponds zalesie kraszeńskie (1) 11.07.2006 nieledew (2) 04.07.06 12.07.06 2 grey heron ardea cinerea l. ciconiformes ardeidae 6 heronries small forest chodlik (2) 2007 dołhobrody (4) 2007 3 mute swan cygnus olor gmel. anseriformes anatidae 5 ponds święcica (1) 02.08.07 garbów (1) 18.07.07 piaski (1) 02.08.07 stary brus (2) 17.07.07 4 coot fulica atra l. gruiformes rallidae 5 ponds garbów 17.07.07 (3) 18.0707 (2) 5 great crested grebe podiceps cristatus l. podicipediformes podicipedidae 5 ponds garbów (1) 11.07.08 samoklęski (2) 31.07.08 stańków (1) 19.07.07 żółtańce (1) 22.07.07 6 black-headed gull larus ridibundus l. charadriformes laridae 7 ponds setters garbów (1) 17.07.07 garbów (3) 11.07.08 garbów (3) 11.07.08 7 common gull larus canus l. charadriformes laridae 1 lake j. wytyckie (1) 15.08.08 8 common tern sterna hirundo l. charadriformes sternidae 5 lake j. wytyckie (5) 15.08.08 9 black tern chlidonias niger l. charadriformes sternidae 1 pond stańków (1) 18.07.07 geophilic dermatophytes 87 lagomorphs lagomorpha, skin, tails and limbs of lizards lacertilia, birds’ wings and feathers (included poultry); chick down; adult birds’ feathers (females moulting during incubation period); chick excrements (the marsh harrier is an altricial bird); less often wings of orthopterans orthoptera or fish scales (dead carp cyprinus carpio). both plant and non-plant components of the nests’ building material were highly compressed because of the birds’ presence in the nests and formed a fixed structure. nests of grey herons (ardea cinerea) were in a heronry in the crowns of tall trees. the base of the nests (a loose cone) was built from thicker sticks; the inner part was built from flexible twigs of deciduous trees: birch (betula sp.), willow (salix sp.) and alder (alnus sp.). the nest lining was sparse, mostly composed of grasses (poacae), various unidentified small roots as well as hair, feathers and fish scales. the lining was absent in some nests. the nest structure was covered with a high amount of white excrements on the inside and the outside as chicks (altricial birds) defecate inside the nest. nests of mute swans (cygnus olor) were recorded on the edge of reed beds of the common reed (phragmites australis) and rushes of the broadleaf cattail (typha latifolia). the nests were stable, large, spherical, 95-161 cm in diameter, 60-140 cm high, at ca. 60 cm immersed in water, wet. the lining was indistinguishable from the rest of the nest due to rotting. the nests were built mostly from rotting parts of the common reed (phragmites australis) and cattails typha sp. as well as willow twigs (salix sp). non-plant material consisted of high amounts of excrements of chicks and adult birds as well as chick eider as chicks spend much time in the nest (although the mute swan is a precocial bird), feathers of adult birds, shells and membranes of hatched eggs (birds do not remove eggshells from the nests). nests of coots (fulica atra) similary to the nests of mute swans were recorded in reed beds of the common reed (phragmites australis) and rushes of the broadleaf cattail (typha latifolia) similarly to the nests of mute swans. they were not very big: table 2 animal matter ratio in the nest structure (in relation to the nest mass) no bird species feathers romains of animal food pellets excrements 1 marsh harrier circus aeruginosus l. h h h h 2 grey heron ardea cinerea l. s s a h 3 mute swan cygnus olor gmel. h a a h 4 coot fulica atra l. s a a s 5 great crested grebe podiceps cristatus l. s, a s, a a s 6 black-headed gull larus ridibundus l. s s a a 7 common gull larus canus l. s s a s 8 common tern sterna hirundo l. s s a s 9 black tern chlidonias niger l. s a a a abbreviation: h – high; s – small; a – absent 88 t. korniłłowicz-kowalska et al. 24-31 cm, spherical, equipped with a type of “pier” providing easier access to water. they were mostly built from stems of broadleaf cattail (typha latifolia) with an addition of sweet flag (acorus calamus), fennel pondweed (potamogeton pectinatus), sporadically containing twigs of black alder (alnus glutinosa) and poplar (populus sp.). the inside of the nests was lined with leaves of common reeds (phragmites australis) and grasses (poacae) poorly distinguishable from the rest of the nest. very small amounts of feathers and faeces in comparison with those recorded in the nests of mute swans were observed. nests of great crested grebes (podiceps cristatus) were recorded on the edge of a phragmites australis reed bed growing on pond banks. they were unstable, floating, quite large, 40-68 cm in diameter, 10-19 cm high, mount-like in shape. most of the nest structure was immersed in water; only a layer of 4-7 cm was above the water surface. the lining was hardly distinguishable from the rest of the nest. the nests were built from rotten plant material in which perennial dicots, a small amount of canadian waterweed (elodea canadensis), rushes (typha latifolia) and fennel pondweed (potamogeton pectinatus) were identified. more permanent plant material consisting of common reeds (phragmites australis) formed the base of the nest. animal material was rarely found in the nest structure: fish scales and bird feathers. excrements were not found (a precocial bird). nests of black-headed gulls (larus ridibundus) nesting in colonies and in setters were recorded in the centre of broadleaf cattail rushes (typha latifolia). the nests were stable, mound-like, quite dry, 27-55 cm in diameter, 7-24 cm high. they were mostly built from remains of aquatic plants, usually broadleaf cattail typha sp. and common reed (phragmites australis). the base was made of twigs of willow salix sp., black alder alnus glutinosa, downy birch betula pubescens, other birches (betula sp.) and european black elderberry sambucus nigra. rhizomes of agropyron sp., reed leaves and water horsetail (equisetum fluviatile) were recorded in the lining which was not always well distinguished from the rest of the nest. very small amounts of feathers and fish scales were found in the nest structure. the nest of common gulls (larus canus) nesting on a lake was recorded on a floating island built from rotten vegetation. the nest was mound-shaped, ca. 30 cm in diameter, consisting of rotten, unidentifiable vegetation. it contained very few feathers and little excrements. nests of common terns (sterna hirundo) were recorded on floating vegetation, mostly water lilies (nymphaea sp. and nuphar sp.). they were also mound-shaped, ca. 20 cm in diameter. they consisted of rotten, unidentifiable vegetation. they contained very few feathers and slightly more excrements than the gull’s nest. the nest of black tern (chlidonias niger) nesting in rushes of typha sp. the nest was a small mound (ca. 20 cm in diameter) made of broken stems of reeds and rushes. the nest did not have any excrements and had very few feathers. isolation and identification of fungi. keratinophilic fungi were isolated with the keratin baiting method using white chicken feathers as the substrate. a total of 390 plates were made. plates were filled with the nest material broken into smaller pieces to ½ and sterile feathers were placed on top. feathers were sterilised using the method of ethylene oxide gassing as described in a study by korniłłowicz (1994). ten plates were prepared from each nest with the exception of nest 2 (circus aeruginosus) from which 20 plates were prepared. nest material ranging from 200 to 500 g was selected geophilic dermatophytes 89 randomly from ten different sites in the nest when the brood chamber was poorly defined or not evident or from its three layers (nest 2, circus aeruginosus) comprising the lining, the outer layer and the layer in between (middle layer) (pugh 1966). plates with the nest material were placed in a humidity chamber and incubated at 26°c for tree to four weeks. the forming mycelium layers were plated onto plates with sabouraud glucose agar with actidion and chloramphenicol obtaining clean fungal cultures by passage. the genus and the species of the fungi were identified using macroscopic characters on plates or microscopic characters in microcultures. preparations of the mycelium developed on feathers were made in a drop of water to identify teleomorphs, which was particularly important for heterotallic species. the fungi were determined using systematic studies by: ellis (1971); domsch et al. (1980); van oorschot (1980); currah (1985); peberdy (1987). determination of physical and chemical properties of the nests. water content in the nest material was determined with the weight method at 105°c. ph in h2o and kcl were measured potentiometrically. total carbon and total sulfur content was determined with an elemental analysis by combustion analysis and in a thermal conductivity detector, c organic content by thiurin method. the content of total n, total p, k, ca, mg was determined after sample mineralisation using the wet assay method in a mixture of concentrated h2so4 and perhydrol using flow spectrophotometry (n-tot., p-tot.) and with the atomic absorption spectroscopy method (k, ca, mg). results assessment. the number of plates (samples) with the nest material showing growth of keratinophilic and non-keratinophilic fungi (an arbitrary term) was used for the general assessment of the occurrence frequency of fungi. it was accepted that one plate can be colonised by only one strain of a fungal species. the species diversity of fungi based on the number of isolates of fungi representing individual species was analysed by calculating simpson’s index (krebs 1994) according to the formula: where pi is the share of isolates (strains) of species „i” in a fungal community and is the quotient of the number of strains of the species and the number of isolates of all fungi obtained on an isolation medium. values of simpson’s index range from 0 to 1-1/s, where s is the number of species in a community of fungi. the species dominance (trojan 1975) was determined using the formula d = 100 . (sa : s) where sa – the sum of isolates of species a, s – the sum of isolates of the group. the group dominance (geophilic dermatophytes and chrysosporium) was determined in a similar way, where sa – the sum of isolates in a group, s – the sum of isolates of all fungi. the following scale was used to assess the frequency of species and groups of keratinophilic fungi: < 1% sporadically; 1-5% rarely; 6-25% frequently; 26-50% very frequently; >50% mass occurrance. correlation coefficients (r) were calculated to define the relationship between the frequency of dominant fungal species and some physical and chemical properties of the nests. 90 t. korniłłowicz-kowalska et al. results physical and chemical properties of the nests. the analysis shows (tab. 3) that the level of humidity was very high and exceeded 80% in the majority of the nests (30 out of 38). lower humidity was recorded only in the nests of grey heron and marsh harrier. a probably secondary increase of humidity was observed in the majority of the nests of grey heron (no 4-8) collected from the ground surface where they had fallen after a storm. the ph of the nests was close to neutral or slightly alkaline table 3 humidity level (in % of dry weight) and ph level in the nests nest no bird species ph humidity h2o kcl 1 marsh harrier 6.95 7.44 70.54 2-i 2-ii 2-iii 7.20 6.70 5.89 7.44 6.81 6.34 76.91 61.42 49.58 3 7.23 6.24 49.56 4 grey heron 7.41 6.49 58.19 5 6.83 5.52 62.3 6 6.47 5.26 50.98 7 5.99 5.46 64.76 8 7.49 6.82 45.67 9 7.76 6.90 18.78 10 mute swan 6.54 6.44 87.72 11 7.10 7.08 82.57 12 6.92 6.90 90.61 13 7.02 6.23 80.72 14 6.55 5.64 78.44 15 coot 6.82 6.86 88.31 16 7.14 7.34 86.71 17 6.70 5.95 84.57 18 7.04 6.99 84.17 19 6.90 6.74 88.47 20 black-headed gull 7.57 7.32 44.09 26 7.15 7.14 85.67 27 7.25 7.04 87.33 28 7.20 7.08 85.54 29 6.84 7.22 86.55 30 6.87 6.78 78.68 31 6.72 6.63 76.21 36 common gull 6.84 6.76 85.26 21 great crested grebe 7.23 6.95 85.34 22 7.10 7.36 86.79 23 7.20 6.85 88.06 24 7.47 6.95 90.11 32 6.90 6.87 79.64 25 common tern 6.47 7.17 82.16 33 black tern 7.15 6.89 85.84 34 7.34 7.05 86.08 35 7.30 7.08 85.02 37 7.10 6.80 85.60 38 6.90 6.63 85.85 abbreviations: 1 – the outer layer of the nest; 2 – the intermediate layer of the nest; 3 – the inner layer (lining) of the nest geophilic dermatophytes 91 (ph in h2o 6.55-7.76) with the exception of one nest of grey heron (no 7) where a weakly acidic ph was recorded (ph in h2o 5.99) (tab. 3). the level of total carbon and organic carbon recorded in the nest material varied (tab. 4). c organic content (in % of dry weight) ranged from 24.1% to 47.9%. total n level was high or sometimes very high ranging from 1.28% d.w. to 5.38% d.w. a high content of total n was particularly high in the nests of grey heron: 2.29%5.38%, which should be attributed to the accumulation of excrements from young birds. different levels of phosphorus and calcium were recorded in the nests. a very high phosphorus content (7.2% d.w.) was observed only in some nests of grey heron. a high level of calcium, as high as 6.92% d.w., was recorded in some nets of mute swan, great crested grebes, black-headed gull and common tern. total s content table 4 the content of some macroelements (in % of dry weight) in the nest material nest number bird species macroelement content (% of the nest dry weight) c total c organic n total s total p total k ca mg 1 marsh harrier 45.68 43.95 2.17 0.28 0.15 0.19 0.70 0.043 2 46.92 42.27 2.27 0.31 0.23 0.40 0.59 0.129 3 47.24 42.44 2.17 0.40 0.09 0.10 0.47 0.030 4 grey heron 36.84 32.30 3.47 0.61 7.20 0.89 6.10 0.309 5 43.94 37.48 2.35 0.40 4.98 0.40 4.08 0.236 6 43.40 35.19 3.06 0.46 6.07 0.36 4.83 0.219 7 41.00 36.02 2.89 0.48 3.50 0.29 5.81 0.187 8 49.10 40.74 2.29 0.34 0.83 0.55 2.03 0.141 9 47.46 39.23 5.38 0.47 0.78 1.19 1.49 0.106 10 mute swan 47.53 43.88 2.72 0.47 0.21 0.53 2.15 0.216 11 37.62 33.18 1.87 0.31 0.20 0.11 6.92 0.105 12 44.93 40.95 2.02 0.33 0.21 0.12 3.32 0.055 13 48.23 45.16 1.79 0.25 0.13 0.37 1.18 0.064 14 41.70 40.19 2.07 0.26 0.11 0.14 0.43 0.040 15 coot 47.66 44.75 1.60 0.28 0.16 0.28 1.27 0.177 16 47.75 43.85 2.53 0.37 0.25 0.40 1.94 0.256 17 45.14 39.92 2.57 0.82 0.18 0.10 1.32 0.249 18 44.82 40.37 2.05 0.31 0.26 1.46 2.13 0.291 19 48.69 45.28 2.19 0.34 0.12 0.11 1.23 0.119 20 blackheaded gull 39.18 34.04 2.55 0.36 0.63 0.47 2.11 0.305 26 43.97 39.92 2.33 0.86 0.20 0.09 2.80 0.212 27 45.36 40.88 2.83 0.91 0.22 0.06 1.98 0.200 28 44.10 40.00 2.80 0.96 0.21 0.09 1.85 0.243 29 46.69 41.72 1.84 0.48 0.15 0.05 1.64 0.169 30 35.09 31.43 2.38 0.42 0.56 0.31 1.57 0.201 31 25.71 24.05 1.39 0.23 0.23 0.52 0.74 0.211 36 common gull 48.88 47.88 2.02 0.29 0.18 0.28 1.99 0.052 21 great crested grebe 42.39 38.25 2.00 0.55 0.10 0.15 3.94 0.060 22 37.47 33.18 2.67 0.52 0.43 0.56 6.42 0.436 23 39.46 34.81 2.00 0.52 0.24 0.23 3.79 0.147 24 41.41 37.11 1.83 0.62 0.17 0.11 1.91 0.111 32 39.55 36.69 1.97 0.29 0.25 0.15 1.08 0.149 25 common tern 46.87 42.52 1.28 0.22 0.15 0.68 1.85 0.185 33 black tern 42.49 40.49 2.83 1.32 0.13 0.07 2.96 0.089 34 44.78 42.40 2.74 1.07 0.11 0.04 2.54 0.073 35 42.46 40.52 2.82 1.20 0.16 0.07 2.88 0.089 37 44.43 41.49 2.62 1.05 0.13 0.05 2.67 0.072 38 41.90 40.57 3.04 1.36 0.16 0.08 3.38 0.092 geophilic dermatophytes 93 the data given in tab. 5 shows that non-dermatophytic fungi representing the chrysosporium group were the dominant group. a total of 273 strains were recorded, which corresponded to 78% of all keratinomycetes identified in the study. the remainder (22%) of the community of keratinophilic fungi was represented by geophilic dermatophytes (75 strains). generic and species diversity of keratinophilic fungi. of the nine genera isolated from the nests, four (microsporum, trichophyton and their teleomorphs nannizzia and arthroderma) represented geophilic dermatophytes and five represented non-dermatophytic fungi: chrysosporium and myceliophthora together with teleomorphs arthroderma, aphanoascus and ctenomyces (tabs 6-8, fig. 2). the genus table 5 numbers of isolated genera, species and isolated of geophilic dermatophytes (gd) and the chrysosporium group (ch) no bird species genus species strain total number of isolates total gd ch total gd ch total gd ch 1 marsh harrier 4 2 2 7 2 5 26 6 20 92 2 grey heron 4 2 2 6 2 5 66 25 41 122 3 mute swan 5 4 1 11 4 7 61 27 34 483 4 coot 7 2 5 10 4 6 52 9 43 282 5 great crested grebe 4 2 2 5 2 3 41 4 37 366 6 black-headed gull 3 1 2 5 1 4 49 4 45 376 7 common gull 8 common tern 2 0 2 3 0 3 53 0 53 472 9 black tern isolated in total 9 4 5 22 9 13 348 75 273 2193 table 6 a list of geophilic dermatophytes and chrysosporium isolated from the nests of wetland birds no species of fungus anamorph teleomorph 1 aphanoascus fulvescens (cooke) apinis + 2 arthroderma cifferii varsavsky et ajello + 3 a. cuniculi dawson + 4 a. curreyi berk. + 5 a. insingulare padhye et carm. + 6 a. quadrifidum dawson et gentles + 7 a. uncinatum dawson et gentles + 8 chrysosporium keratinophilum frey ex carm. + 9 ch. pannicola (corda) van oorschot et stalpers + 10 ch. quenslandicum apinis et rees + 11 ch. tropicum carm. + 12 myceliophthora an. arthroderma tuberculatum kuehn + 13 chrysosporium an. a. curreyi berk. + 14 chrysosporium an. renispora flavissima sigler et al. + 15 ctenomyces serratus eidam + 16 microsporum cookei ajello + 17 m. fulvum uriburu + 18 m. gypseum (bodin) guiart et grigoriakis + 19 myceliophthora an. ct. serratus + 20 nannizzia gypsea (nannizzi) stockdale + 21 trichophyton ajelloi (vanbr.) ajello + 22 t. terrestre durie et frey + abbreviations: (+) yes; (-) no geophilic dermatophytes 95 the distribution of keratinophilic fungi and nest properties. data on the occurrence of individual species of geophilic dermatophytes and chrysosporium in each of the 38 nests are presented in tables 7 and 8. they show a non-uniform distribution of the populations of keratinophilic fungi in the microhabitat: their occurrence was observed in some nests while they were absent in others. this corresponded to table 7 the frequency and distribution of individual species of geophilic dermatophytes in the nests of wetland birds nest number bird species fungal species totala.i. a. q. a. u. m. c. m. f. m. g. n.g. t. a. t. t. 1 marsh harrier 0 2 0 3 4* 2 6 4 grey heron 7 11 5 0 6 0 7 0 8 5 7 12 9 6 6 10 mute swan 0 11 1 4 5 12 1 1 2 13 7 3 10 14 10 10 15 coot 2 1 3 6 16 1 1 17 1 1 2 18 0 19 0 20 blackheaded gull 4 4 26 0 27 0 28 0 29 0 30 0 31 0 36 common gull 0 21 great crested grebe 0 22 3 3 23 1 1 24 0 32 0 25 common tern 0 33 black tern 0 34 0 35 0 37 0 38 0 in total 5 5 3 2 1 22 3 1 33 75 abbreviations: a.i. – arthroderma insingulare; a.q. – a. quadrifidum; a.u. – a. uncinatum; m.c. – microsporum cookei; m.f. – m. fulvum; m.g. – m. gypseum (complex); n.g. – nannizzia gypsea; t.a. – trichophyton ajelloi; t.t. – t. terrestre (complex); * – number of strains 96 t. korniłłowicz-kowalska et al. different physical and chemical properties of the nests, primarily the humidity level and ph. in the group of geophilic dermatophytes, trichophyton terrestre together with the perfect stages: arthroderma insingulare and a. quadrifidum, mostly colonised the nests of grey herons and single nests of marsh harriers and black-headed gulls (tab. 7). their humidity was lower than that observed in other nests (tab. 3) and ranged table 8 the frequency and distribution of species of the chrysosporium group in the nests of wetland birds nest number bird species fungal species a.f. a. cif. a. cun. a. cur. ch. k. ch. p. ch. q. ch. trop. ch. tub. ch. cur. ch. fl. ch. sp. m. ser. c. ser. total 1 marsh harrier 6* 8 3 17 2 0 3 2 1 3 4 grey heron 5 7 1 1 14 5 10 10 6 4 5 9 7 4 4 8 1 1 9 2 1 3 10 mute swan 2 1 1 3 3 10 11 1 1 12 5 3 5 1 14 13 4 4 14 4 1 5 15 coot 3 1 3 7 16 4 7 11 17 2 4 2 1 1 10 18 4 1 1 3 3 12 19 1 2 3 20 blackheaded gull 2 1 1 4 26 8 8 27 2 5 1 6 28 5 5 29 0 30 10 10 31 0 36 common gull 10 10 21 great crested grebe 10 10 22 9 1 9 23 2 2 4 24 2 4 6 32 7 7 25 common tern 8 1 9 33 black tern 8 8 34 7 7 35 7 7 37 3 10 10 38 9 9 in total 25 9 2 6 159 8 7 40 1 1 3 2 5 5 273 abbreviations: a.f. – aphanoascus fulvescens; a.cif. – arthroderma cifferii; a.cun. – a. cuniculi; a.cur. – a. curreyi; ch.k .– chrysosporium keratinophilum; ch.p. – ch. pannicola; ch.q. – ch. queenslandicum; ch.trop. – ch. tropicum; ch.tub. – ch. tuberculatum; ch. cur. – ch. curreyi an arthoderma curreyi; ch.fl. – chrysosporium an. renispora flavissima; ch.sp.– chrysosporium sp.; m.ser. – myceliophthora an. ctenomyces serratus; c.ser. – ctenomyces serratus; * – number of strains geophilic dermatophytes 97 from 18.78% to 62.30%, and their ph was alkaline (ph in h2o 7.32-7.76). another dermatophyte species recorded more frequently, microsporum gypseum together with the teleomorph nannizzia gypsea, colonised only nests of mute swans (three of the five studied) and coots (two of the three studied). these nests had a neutral ph: ph in h2o 6.55-7.10 (tab. 3). the most numerous species within both communities of keratinophilic fungi, chrysosporium keratinophilum together with its teleomorph aphanoascus fulvescens, showed preferences for habitats characterised by a very high humidity, which was recorded in the case of coot’s nests, great crested grebe’s nests as well as the nests of both species of gulls and terns (tab. 8). apart from the above species, chrysosporium tropicum, one of more frequent species, mostly colonised grey heron’s nests while its occurred rarely or sporadically or did not occur at all in others (tab. 8). it was shown that the frequency of t. terrestre (together with the teleomorph) is negatively correlated with the nest’s humidity level and that of ch. keratinophilum is positively correlated with it. an even stronger and positive correlation was observed between the frequency of occurrence of t. terrestre and the nest’s ph and phosphorus content. the frequency of both species: ch. keratinophilum (together with the teleomorph) and t. terrestre (together with the teleomorph), was also significantly positively correlated with the calcium content in the nests although the correlation coefficients were lower than those for ph and phosphorus content (tab. 10). the colonisation rate of the nests and the species composition of so-called nonkeratinophilic fungi growing on feathers. the colonisation rate of the nest material by ubiquistic fungi (polyphages), arbitrarily called non-keratinophilic, varied greatly and ranged from 12% to 95%. the greatest number of ubiquistic fungi able table 10 correlation coefficients between the frequency of selected species of keratinophilic fungi (together with the teleomorph) and some physical and chemical properties of the nest material (p=0.05) no property chrysosporium keratinophilum + teleomorph trichophyton terrestre + teleomorph microsporum gypseum + teleomorph 1 humidity 0.62 0.61 0.28 2 ph h2o 0.28 0.81 0.41 3 total nitrogen content 0.324 0.819 -0.37 4 phosphorus content 0.08 0.97 0.22 5 calcium content 0.63 0.54 0.03 table 9 simpson’s index of species diversity (d) for communities of keratinophilic fungi in the nests no bird species simpson’s index 1 marsh harrier 0.8018 2 grey heron 0.6942 3 mute swan 0.8400 4 coot 0.8300 5 great crested grebe 0.4400 6 gull (black-headed + common) 0.3900 7 tern (common + black) 0.1400 98 t. korniłłowicz-kowalska et al. table11 a list of non-keratinophilic fungi isolated from the nests of wetland birds with the keratin baiting method no fungal species 1 2 3 4 5 6 7 8 9 total 1 acremonium chrysogenum (thirum and sukap) w. gams 5* 5 2 a. rutilum w.gams 1 1 3 acremonium sp. 1 1 4 alternaria alternata (fr.) keissler 4 1 5 5 aspergillus flavus link ex gray 5 5 6 a. fumigatus fres. 7 9 2 1 4 3 2 1 29 7 a. niger van tieghem 1 1 8 candida sp. 1 1 9 chaetomium globosum kunze ex fr. 10 10 10 doratomyces microsporus (sacc.) morton & g.sm. 6 1 7 11 d. nanus (ehrenb. ex link) morton & smith 1 1 12 fusarium culmorum (schwabe) snyd. et hansen 1 1 13 f. equiseti (corda) sacc. gordon 1 1 14 f. solani (mart.) sacc. 1 1 15 geotrichum candidum link ex leman 1 1 16 geotrichum sp. 1 1 2 17 gliocladium catenulatum gilm. & abbott 1 2 10 13 18 g. roseum bain. 1 1 2 19 mariannea elegans (corda) samson 2 2 20 monocillium indicum saksena 1 1 21 paeciliomyces farinosus (holm ex gray) a.h.s. brown & g. sm. 1 1 22 p. lilacinus (thom) samson 1 1 1 3 23 p. variotii bain. 1 1 24 papulaspora immersa hotson 1 1 25 penicillium chrysogenum thom 2 2 26 p. expansum link ex gray 1 1 1 3 27 p. purpurogenum stoll 4 2 6 28 p. verrucosum dierckx 5 2 7 29 penicillium sp. 1 3 1 5 30 scedosporum apiospermum (sacc.) sacc. ex castell & chalmes 1 1 31 scopulariopsis acremonium delacr. vuill. 1 1 32 s. brevicaulis (sacc.) bain. 6 10 2 1 1 6 1 27 33 torula herbarum pers. ex gray 1 1 34 trichoderma viride pers. ex gray 1 1 3 1 6 35 trichothecium reseum (pers.) link ex gray 1 1 36 verticillium chlamydosporum goddard 1 1 geophilic dermatophytes 99 to colonise feathers occurred in the nests of grey herons and marsh harriers, and the smallest number was recorded in the nests of great crested grebes and mute swans (fig. 1). unlike keratinophilic fungi, the colonisation rate of native keratin by ubiquistic fungi corresponded to the richness and frequency of their species (fig. 1, tab. 11). the greatest richness and species diversity was observed in the case of non-keratinophilic fungi colonising the nests of grey herons and marsh harriers: 19 and 11 species and 50 and 39 strains, respectively (tab. 11). the smallest species differentiation of the biota of ubiquistic species colonising feathers was observed in the nests of both tern species: common tern and black tern, 5 and 3, respectively, represented by single strains (tab. 11). as regards the species composition, ubiquistic fungi recorded in the nests and colonising native feathers were represented by 34 species belonging to 20 genera (the species of ten isolates was not determined). the most frequently isolated genera were aspergillus, gliocladium, paeciliomyces, penicillium and scopulariopsis (tab. 11). similarly to keratinophilic species, individual species of non-keratinophilic fungi showed preferences for nests of specific bird species. scopulariopsis brevicaulis was most frequently isolated from the nests of grey herons and marsh harriers: 15% and 20%, respectively, and aspergillus fumigatus: an 18%-share within nonkeratinophilic fungi colonising these nests. additionally, doratomyces microsporus was frequently isolated from marsh harrier’s nests on feathers (15% respectively). among other ubiquistic species, two polyphagous species: gliocladium catenulatum and verticillium lecani, occurred as co-dominant species in black-headed gull’s nests: the colonisation rate of feathers was 28% and 25%, respectively. on the other hand, chaetomium globosum, which represented ca. 63% of total non-keratinophilic fungi, was an accompanying species of feather colonisation by typically keratinophilic fungi in mute swan’s nests (tab. 11). discussion the present study shows that keratinophilic fungi colonised 86.8% of the nests of wetland birds. a slightly higher (ca. 5%) occurrence frequency of keratinomycetes was recorded only in nest boxes (hubalek et al. 1973). the occurrence frequency of keratinophilic fungi, however, was higher in comparison with open-cup nests of land fungi. hubalek et al. (1973) demonstrated the presence of keratinophilic fungi in 72.7% of such nests, mostly belonging to passeriformes. 37 v. lecani (zimm.) viegas 3 1 5 9 1 19 38 v. psalliotae terschow 4 4 1 1 10 39 verticillium sp. 2 2 40 yeasts 2 1 1 4 total 39 50 16 27 14 35 0 6 5 192 abbreviations: 1 – marsh harrier; 2 – grey heron; 3 – mute swan; 4 – coot; 5 – great crested grebe; 6 – black-headed gull; 7 – common gull; 8 – common tern; 9 – black tern; ( )* – number of strains 100 t. korniłłowicz-kowalska et al. the investigations also showed a high (76% on average) colonisation rate of the nest material by keratinophilic fungi. the nest material of marsh harriers (49%) was the least colonised and that of both tern species (90%) was the most strongly colonised material. the widespread distribution of keratinophilic fungi in the nests of wetland birds was conditioned by the presence of the birds (breeding) and keratin matter, mostly feathers and less frequently hair, animal food remains, excrements and pellets. a considerable accumulation of total nitrogen as well as phosphorus and calcium indicated nest contamination with remains of animal origin. both the nutrient factor (keratin) and high humidity as well as neutral to alkaline ph (ph 6.5-7.8) of the nest material were favourable for the development of keratinophilic fungi in the nests (tab. 2). as previous investigations show (korniłłowicz-kowalska 1997), keratinophilic fungi grow well on surfaces of feathers which are a non-wettable substrate when such substrate is in contact with water. the process is intensive when the substrate’s ph ranges between 6.5 and 7.8 (korniłłowicz-kowalska, bohacz 2002), which is connected with the optimum of extracellular keratinolytic proteases of these fungi (korniłłowicz-kowalska 1999). similar observations were made by kunert (2000) in relation to biodegradation of hair by keratinolytic fungi. on the whole, a high richness of kertinomycete species was observed in the nests of wetland birds examined: altogether 22 species belonging to nine genera were recorded. a total of no more than 15 species of keratinomycetes is regularly isolated from natural environments such as the soil (gueho, villard and guinet 1985; korniłłowicz-kowalska, bohacz 2002). however, a high differentiation of the composition and the frequency of keratinomycete species colonising the nests was observed in the investigations depending on the species of the nesting bird. the greatest number of fungal species and their diversity, was observed in the nests of mute swans and coots and, further, of march harriers and grey herons. the smallest number of species and the lowest simpson’s indices were recorded in the nests of both tern species and black-headed gulls. great differences in the species composition of keratinophilic fungi in nests depending on the bird’s species were previously demonstrated by hubalek (1974) in his analysis of terrestrial birds, mostly passeriformes. it is interesting that the nests in which the greatest richness and diversity of keratinomycete species were observed (mute swans and coots) differed considerably by keratin matter content (feathers). high amounts of feathers and excrements were observed in mute swan’s nests while small amounts were noted in the coot’s nests or they were absent (tab. 2). on the other hand, the two birds’ species had similar breeding biotopes and feeding grounds. both colonized fertile reservoirs (ponds), built nests in reed beds and broadleaf cattail rushes, and mostly fed in the littoral zone (mute swans also in the middle of the ponds), feeding on vegetation and small invertebrates (snails, insects) occurring on plants and the bottom slime of shallow waters. a high rate of contamination of the nests by geophilic keratinophilic fungi may also have been connected with the contamination of the feeding grounds of these birds by the fungi. a high accumulation of geophilic keratinophilic fungi is observed in bottom sediments and reservoir waters affected by strong anthropopressure such as ponds (korniłłowicz 1993; ulfig 1986, 1987; ulfig, ulfig 1990; ulfig et al. 1996). allochthonic organic substances, including keratin remains (feathers, hair, geophilic dermatophytes 101 etc.), are a source of these fungi (korniłłowicz 1993; ulfig et al. 1996). it is highly probable that the fungi may have been mechanically transferred on the plumage or collected by the birds with the food. the fact that the frequency and the distribution of keratinophilic fungi on the surface of birds’ bodies depends on their feeding habitats was previously reported by pugh (1965, 1966). bird excrements may also have been a source of keratinophilic fungi in the nests of many bird species. faeces contamination was observed especially in the case of grey herons, mute swans and marsh harriers (tab. 2). the occurrence of keratinophilic fungi in excrements and their spread by excretion with faeces have been reported by, e.g., dominik, majchrowicz (1970); nooruddin, singh (1987); garetta et al. (1992). prey remains were also an important contribution to the keratinolytic mycobiota in the nests of marsh harriers and grey herons: birds, small mammals (marsh harriers), hair (grey herons), and pellets (both species). bird plumage, mammal hair, skin scales and pellets are obviously colonised by different species of geophilic dermatophytes and chrysosporium (pugh 1966; rees 1967; pugh, evans 1970; hubalek et al. 1973; hubalek 1974; sur, ghos 1980; korniłłowicz-kowalska, kitowski 2009). the thesis that feeding habitats and “animalisation” (enrichment in keratin remains) are mostly a source of keratinophilic fungi in the nests of wetland birds is also corroborated by the observations of breeding biotopes and feeding grounds of other bird species examined in the study. few species of keratinophilic fungi were recorded in the nests of these birds (great crested grebe, black-headed gull, common tern and black tern). bird feathers or excrements were also observed in them sporadically. apart from black terns, these birds are piscivorous, search for fish in the water, diving into it (great crested grebe) or catching fish from the air (blackheaded gull, common tern). black terns, on the other hand, are insectivores and catch insects in flight, over the water surface and fields. this manner of feeding and the types of feeding sites (in the water, air above the reservoir) are not favourable for the acquisition by birds of geophilic keratinophilic fungi that can occur in these environments only accidentally. the majority of bird species examined did not come into contact with the soil (an environment believed to be a major reservoir of geophilic keratinophilic fungi) or such contact was rare. marsh harriers which often hunt outside the breeding site, in meadows and fields, catching small mammals, lagomorphs, sometimes poultry, and grey herons which supplement their diet with voles outside the breeding season, were the only exceptions. little importance of the soil as a source of contamination of the plumage and nests of the majority of fungi was previously reported by pugh (1966) and rees (1967). our examinations show that non-dermatophytic keratinophilic fungi of the chrysosporium group are a dominant group in the nests of wetland birds. they represented ca. 78% of the keratinophilic mycobiota of the nests (273 nests), while the genus chrysosporium itself constituted ca. 63%. the dominance of chrysosporium in the nests of land birds was reported by hubalek (1974) and hubalek et al. (1973) several times. in their analysis of the nests of passerines, mostly eurasian tree sparrow passer montanus, hubalek et al. (1973) showed that “chrysosporia” constituted over 90% of keratinomycete populations in the nests. dermatophytes ranged only from ca. 2% to ca. 9% of the fungi (hubalek et al. 1973). the share of geophilic 102 t. korniłłowicz-kowalska et al. dermatophytes was between 0% (both species of terns and gulls) and 44.3% (mute swan) in the nests of wetland birds studied. a generally higher frequency of chrysosporium in comparison with geophilic dermatophytes in birds’ nests may be connected with their higher occurrence in the plumage and on birds’ feathers (hubalek 2000) and a lower keratinolytic activity (korniłłowicz-kowalska 1997; kunert 2000). due to the latter, these fungi grow better in environments containing more accessible keratin sources, such as feather keratin rather than, for instance, hair keratin. moreover, nest ph (ph in h2o 6.5-7.8) was a factor favourable for a high frequency of chrysosporium in the nests of wetland birds examined. the majority of chrysosporium species prefer environments with a higher ph and are alkalotolerant (kushwaha 2000). ch. keratinophilum was the most frequently isolated species from the nests of wetland birds. together with its telefomorph (aphanoascus fulvescens), it colonised the nests of all the birds and its share in the community of keratinophilic fungi was 53% on average. ch. tropicum (11.5% of total keratinomycetes), isolated mostly from the nests of grey herons, was less widespread. according to hubalek (1974), a. fulvescens mostly colonises nests of wetland birds and ch. tropicum is a frequent coloniser of these nests. trichophyton terrestre, which together with its teleomorphs (arthroderma quadrifidum and a. insingulare) constituted 12.5% of all isolated fungi, and microsporum gypseum and its telefomorph (nannizzia gypsea), which constituted 7.1%, had the highest frequency among geophilic dermatophytes. populations of t. terrestre mostly colonised the nests of marsh harriers and grey herons while m. gypseum colonised the nests of mute swans. the frequency and distribution of individual keratinomycete populations in the nests of wetland birds was conditioned primarily by the differences in the humidity and ph level of the nests. a similar phenomenon was observed in a study on the frequency of keratinomycetes in the soil by chmel et al. (1972) as well as by korniłłowicz (1993) and korniłłowicz-kowalska, bohacz (2002). soil ph was the most important selection factor in the populations of these fungi (korniłłowicz 1993; korniłłowicz-kowalska, bohacz 2002). the present investigations show that a high humidity of the nest material was the reason for the accumulation of ch. keratinophilum in the nests of wetland birds. the occurrence frequency of the fungus increased as water content increased (r=0.62, p=0.05). the colonisation of nests with a high humidity level (ok. 62%) by ch. keratinophilum was also observed by hubalek et al. (1973), who reported that ch. keratinophilum (as a teleomorph) is isolated more frequently from the plumage of water birds than land birds. ch. keratinophilum’s preference for environments with a high level of humidity results from its hygrophilous (hydrotolerant) nature, which is related to a high demand for water (garg et al. 1985; hubalek 2000). ch. keratinophilum is also an alkalotolerant species. a reverse relationship with the humidity level in the nests was observed in the population of trichophyton terrestre, a species belonging to xerophyles (garg et al. 1985). the frequency of occurrence of this dermatophyte decreased together with an increase in the water content in the nest material (r=-0.61, p=0.05). t. terrestre’s preference for dry environments was also observed by chmel et al. (1972) and chmel & vláčiliková (1975). geophilic dermatophytes 103 the present investigations also confirm growth stimulation of t. terrestre in alkaline environments previously observed by other authors (chmel et al. 1972; ulfig et al. 1996). the frequency of this dermatophyte increased as ph increased (r=0.81, p=0.05) reaching its maximum in the nests of grey herons in which ph in h2o was ca. 7.4-7.8. in the case of t. terrestre, a high content of n-total, phosphorus and calcium was also a factor significantly conditioning its frequency of occurrence in the nests. it may be supposed that a high level of these elements contributed primarily to a ph increase in the nests. nest alkalisation was caused by the release of ammonia produced during the ammonification of uric acid contained in bird faeces (accumulated in very high amounts in the nests of grey herons) and calcium ions and phosphates from the digestion of animal food (fish) and excreted in faeces. the relationship between the frequency of t. terrestre and the calcium level in the environment (soil) was reported by chmel et al. (1972). microsporum gypseum is also interesting in the group of other, more frequently isolated species of keratinophilic fungi. although no significant correlations between its frequency of occurrence (the number of samples with fungal growth was too small) and the physico-chemical properties of the nests were observed, its occurrence limited mostly to the nests of mute swans and coots may suggest preferences for environments with neutral ph and a relationship with biotopes polluted with organic matter. m. gypseum is a dominant dermatophyte species in bottom sediments of waters strongly polluted by communal waste waters delivering considerable amounts of keratin matter (ulfig 2000). similar observations were made on the occurrence of m. gypseum in soils polluted with waste waters (ali-stayech, jamous 2000). previous analyses (korniłłowicz-kowalska, bohacz 2002) of the occurrence and the distribution of geophilic dermatophytes and chrysosporium in soils with different physico-chemical properties showed that m. gypseum (as well as m. cookei) colonises exclusively soils characterised by a considerable “animalisation” and neutral ph. it should be stressed that the majority of keratinomycete species recorded more frequently in the nests of wetland birds were thermotolerant fungi such as ch. keratinophilum, ch. tropicum, m. gypseum. they grow well at a temperature of 37°c and maximum growth temperatures are 40-41°c (garg et al. 1985). this is consistent with nest temperatures during incubation reaching a maximum of 40-41°c (pinowski et al. 1999). on the other hand, species considered to be typical soil species such as trichophyton ajelloi and ctenomyces serratus (domsch et al. 1980) were rare in the nests. neutral or alkaline ph of the nests of wetland birds did not encourage the occurrence of acidophilic species such as t. ajelloi, arthroderma uncinatum, a. curreyi (garg et al. 1985). the exception was chrysosporium tropicum, which is thought to be acidophilic according to garg et al. (1985). it occurred relatively frequently in the nests of grey herons where ph (h2o) was from 5.99 to 7.76. hubalek et al. (1973) also reported the occurrence of ch. tropicum in nests with ph ranging from acidic of slightly alkaline (ph 5.5-7.5). it is possible that the species colonises biotopes with a broad ph range, adapted mostly to high temperatures (van oorschot 1980) and resistant to light. extreme conditions are observed in grey heron’s nests during breeding: low humidity, sun exposure and additional nest heating related to it. this allows only species of keratinomycetes, such as t. terrestre and ch. tropicum, that are most resistant to the lack of water and insolation, to survive. 104 t. korniłłowicz-kowalska et al. of the species of keratinophilic fungi recorded in the nests of wetland birds, a widespread occurrence and a high frequency of ch. keratinophilum in many nests and the accumulation of m. gypseum in the nests of mute swans may raise greatest concerns. ch. keratinophilum and its teleomorph aphanoascus fulvescens are opportunistic non-dermatophitic causal agents of mycoses in humans and animals (gueho et al. 1985). microsporum gypseum is the most virulent geophilic dermatophyte causing inflammatory tinea corporis and tinea capitis in humans (hayashi, toshitani 1983; offidani et al. 1998). it also causes mycoses in animals (garetta et al. 1992). the results obtained in this study also confirmed the occurrence of populations of potentially pathogenic ubiquistic moulds, aspergillus fumigatus and scopulariopsis brevicaulis, in the nests, observed in a previous study (korniłłowicz-kowalska, kitowski 2009). the species are thermotolerant and alkalotolerant, and show keratinolytic abilities (kozakiewicz, smith 1994; santos et al. 1996; filipello-marchisio et al. 2000). pathogenic strains of these fungi cause lung aspergillosis (a. fumigatus) and onychomycosis (s. brevicaulis) (dvořak, otčenašek 1969). aspergillus fumigatus is very frequently isolated from nests of birds, their ontocoenoses, plumage and pellets (hubalek 1974; kruszewicz et al. 1995; shin et al. 1996; korniłłowicz-kowalska, kitowski 2009). it is the most frequent causal agents of mycoses, mostly of the lungs and air sacs in wetland birds (mikaelian et al. 1997). the present investigations show that nests are a potential source of pathogenic infections with a. fumigatus in wetland birds. they are also some of reservoirs of geophilic fungi causing dermatomycoses and systemic mycoses in humans and mammals. these fungi can penetrate the water body from the nest and can be transferred by birds over considerable distances during migration. acknowledgement. this work was supported by the polish committee of scientific researches, grants no. 2p04g03330. references ali-stayech, jamous r.m.f. 2000. keratinophilic fungi and related dermatophytes in polluted soil and water habitats. 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(japan) 22: 347–352. trojan p. 1975. ekologia ogólna. wyd. pwn, warszawa. ulfig k. 1986. dermatofity w osadach dennych i osadach ściekowych. post. dermatol. 3: 385–389. ulfig k. 1987/1990. grzyby keratynofilne w osadach dennych wód powierzchniowych. acta mycol. 23: 3–11. ulfig k. 2000. the occurrence of keratinolytic fungi in waste and waste-contaminated habitats. (in:) r.k.s., kushwaha, j. guarro (eds). biology of dermatophytes and other keratinophilic fungi. rev. iber. micol. 17: 44–50. ulfig k., lukasik w., plaza g. 1996. further statistical evaluation of the occurrence of keratinolytic fungi in dam sediments. intern. j. environ. health res. 6: 39–47. ulfig k., ulfig a. 1990. keratinophilic fungi in bottom sediments of surface waters. j. med. vet. mycol. 28: 419–422. geophilic dermatophytes 107 dermatofity geofilne oraz inne grzyby keratynofilne w gniazdach ptaków wodno-błotnych streszczenie w prezentowanej pracy zbadano częstość występowania oraz skład gatunkowy grzybów keratynofilnych w 38 gniazdach 9 gatunków ptaków wodno-błotnych. gniazda zbierano w latach 2006-2009 na terenie lubelszczyzny (płd.-wsch. polska), po opuszczeniu przez ptaki. posługując się metodą przynęty keratynowej przebadano 390 próbek materiału gniazdowego, uzyskując 348 szczepy keratinomycetes. obecność grzybów keratynofilnych stwierdzono w 86,8% badanych gniazd. były one reprezentowane przez 9 gatunków dermatofitów geofilnych oraz 13 gatunków chrysosporium. 78% wyosobnionych szczepów stanowiły grzyby z grupy chrysosporium, 22% należało do dermatofitów geofilnych. gatunkiem dominującym był ch. keratinophilum, który wraz z teleomorfą (aphanoascus fulvescens) reprezentował 53% keratynolitycznej mykobioty badanych gniazd. do mniej licznych ale częstych należały populacje: ch. tropicum (11,5%), trichophyton terrestre wraz z teleomorfą (12,5%) oraz microsporum gypseum włącznie z teleomorfą (7,1%). wyselekcjonowane populacje reprezentowały głównie grzyby termotolerancyjne, preferujące środowiska o odczynie obojętnym i alkalicznym (ch. keratinophilum, a. fulvescens, t. terrestre, m. gypseum) oraz o dużej wilgotności (ch. keratinophilum). przeprowadzone badania wykazały, że gniazda ptaków wodno-błotnych cechują się wysoką frekwencją takich gatunków grzybów potencjalnie chorobotwórczych dla ptaków, ssaków oraz człowieka jak m. gypseum, ch. keratinophilum i aspergillus fumigatus. 2014-01-01t11:52:01+0100 polish botanical society 2014-09-06t20:24:34+0200 polish botanical society 2014-08-28t17:48:59+0200 polish botanical society 2014-08-31t21:55:04+0200 polish botanical society 2014-08-25t14:57:10+0200 polish botanical society © the author(s) 2014 published by polish botanical society leccinum variicolor (basidiomycota, boletales) in poland małgorzata stasińska and zofia sotek department of botany and nature conservation, university of szczecin felczaka 3c, pl-71-412 szczecin; stasinsk@univ.szczecin.pl stasińska m., sotek z.: leccinum variicolor (basidiomycota, boletales) in poland. acta mycol. 49 (1): 69–78, 2014. leccinum variicolor watling is widespread but it is a rare species in poland and some other countries, and is either protected or red-listed. this paper describes the morphology and ecology of l. variicolor as well as its distribution in poland. key words: macrofungi, threatened fungi, distribution, ecology introduction the genus leccinum gray of the family boletaceae chevall. and the order boletales e.-j. gilbert is represented by ca. 75 species (kirk et al. 2008). fourteen of them are known from poland (wojewoda 2003; łuszczyński 2008). almost all species of this ectomycorrhizal genus are generally highly host-tree specific (den bakker et al. 2004). leccinum variicolor is widespread, but not common in europe, and occurs in the northern hemisphere, in the temperate, boreal, subalpine and arctic zones (e.g. den bakker, noordeloos 2005; knudsen, taylor 2012; noordeloos (online)). it is a rare species in poland although it is known from various regions in the country, e.g. from the lublin region (flisińska 2004), the wielkopolska region (ślusarczyk 2007) and the góry świętokrzyskie mts (łuszczyński 2007, 2008). leccinum variicolor is included on the red list of macrofungi in upper silesia (wojewoda 1999) and red list of basidiomycetes in the góry świętokrzyskie mts (łuszczyński 2002, 2008). this paper describes the morphology and ecology of leccinum variicolor as well as its distribution in poland. material and methods the description of the basidiocarp morphology and habitats is based on original specimens and their localities, supplemented by information from the literature. acta mycologica vol. 49 (1): 69–78 2014 doi: 10.5586/am.2014.006 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 70 m. stasińska and z. sotek © the author(s) 2014 published by polish botanical society microscopic features were measured by standard light microscopy (lm). basidia and spore measurements in the descriptions of the species are based on 30 measurements per specimen from 18 collections. dimensions of the basidia and the spores are given as follows: (minimum value-) 1st decile – 9th decile (-maximum value). spore length to width ratios are reported as q. micrographs were taken with a scanning electron microscope zeiss evo ls10 (sem) in the centre for molecular biology and biotechnology, environmental testing laboratory university of szczecin (poland). the cartogram map (fig. 1) is based on our investigations and all available published data and unpublished (e.g. herbarium) records. the distribution was mapped using the grid square system following the “atlas of the geographical distribution of fungi in poland” (wojewoda 2000). geographical regions are given after kondracki (2002). the nomenclature of vascular plants follows mirek et al. (2002) and that of plant communities follows matuszkiewicz (2001). the specimens were collected by the authors in pomerania during 1998-2011 and were deposited in the herbarium of the department of botany and nature conservation, szczecin university (szub), poland. results leccinum variicolor watling, notes r. bot. gdn edinb. 24: 268 (1969) – boletaceae, boletales, agaricomycetidae, agaricomycetes, agaricomycotina, basidiomycota, fungi (kirk et al. 2008). fig. 1. distribution of leccinum variicolor in poland, in atpol grid square system (wojewoda 2000): 1 – data published, 2 – data unpublished. leccinum variicolor (basidiomycota, boletales) in poland 71 © the author(s) 2014 published by polish botanical society syn.: boletus variicolor (watling) hlaváček; krombholziella variicolor (watling) šutara; leccinum oxydabile (singer) singer ss. kreisel, pilát & dermek, singer p.p.; leccinum variicolor var. bertauxii lannoy & estadès; for other synonymies see index fungorum. pileus 30-120 mm in diameter, convex to plano-convex, with entire margin, dark brown to blackish brown, greyish brown or mouse grey, usually with lighter ochraceous or yellowish grey spots, tomentous; tubes ventricose, 7-18 mm long, white when young, later grey or cream, vinaceous to brown when bruised; pores ca. 0.5 mm in diameter, white or cream, ochraceous when bruised; stipe 70-180 x 10-35 mm, solid, cylindrical to clavate, white, with small grey to blackish brown squamules, often discolouring greenish blue at base; context white, gradually or quickly staining pink to coral red in pileus and apex of upper half of stipe, often intensely green blue in the lower half of stipe after 1-5 hours (fig. 2); spores (10) 13.5-17.5 (20.0) x 5.06.5 (7.0) µm, q = 2.4-3.1, smooth, fusiform with or without a suprahilar depression (fig. 3a, b); basidia 23.0-32.0 (34.2) x 8.5-11.0 (13.0) µm, mostly with 4 sterigmata. habitat and distribution. leccinum variicolor (mottled bolete) is a mycorrhizal species, closely associated with tree species of the genus betula spp. (den bakker et al. 2004, 2007; legon et al. 2009). it mainly grows on humid soil, peat or among sphagnum spp., often in waterlogged sites. it occurs in mesic deciduous, coniferous and mixed forests, in open sites and in peatlands (e.g. siller et al. 2005; legon et al. 2009; chinan 2011; knudsen, taylor 2012; noordeloos (online)). in poland, it has usually been noted in raised transitional bogs and their margins (e.g. ślusarczyk 2007; kujawa, gierczyk 2011; stasińska 2011). to date it has been recorded in associations such as sphagno squarrosi-alnetum (halama, romański 2010), vaccinio uliginosi-betuletum pubescentis and vaccinio uliginosi-pinetum (flisińska 1987 (1988), 2004; stasińska 2011) and communities such as abies alba-sphagnum girgensohnii (łuszczyński 2007) and betula pendula-betula pubescens (ślusarczyk 2007). it has recently been recorded in erico-sphagnetum medii, myrico-salicetum auritae and the community eriophorum vaginatum-sphagnum fallax (grzesiak 2010, unpublished). leccinum variicolor is widespread and its range is probably circumboreal (knudsen, taylor 2012; noordeloos (online)). it is known in north america (canada, usa), asia (china, japan) and europe from, e.g., austria, estonia, france, germany, great britain, macedonia, hungary, the netherlands, romania, switzerland, and scandinavian countries (krieglsteiner 1991; lannoy, estades 1995; den bakker, noordeloos 2005; siller et al. 2005; fu et al. 2006; kalamees, saar 2006; den bakker et al. 2007; karadelew et al. 2007; legon et al. 2009; chinan 2011; knudsen, taylor 2012). in poland leccinum variicolor is known from 30 localities. published data are available for twelve sites (flisińska 1987 (1988); sałata 1988; ślusarczyk 2007; łuszczyński 2008; gierczyk et al. 2009; halama, romański 2010; kujawa, gierczyk 2011, 2013; stasińska 2011). the majority of localities are in southern poland, chiefly in the niziny środkowopolskie plains and in the wyżyna małopolska upland. it has been recorded sporadically in other parts of poland (fig. 1). localities in poland. specimens indicated by asterisk (*) have not been examined by the authors. ac-89 – pobrzeża południowobałtyckie littoral regions: gdańsk-owczarnia (trójmiejski landscape park); peat bog, among sphagnum, 03.10.2009, leg. et det. m. wantoch-rekowski (unpublished, grej 2013)*. 72 m. stasińska and z. sotek © the author(s) 2014 published by polish botanical society fig. 2. fruit-bodies of leccinum variicolor recorded in roby reserve (photo by m. stasińska). leccinum variicolor (basidiomycota, boletales) in poland 73 © the author(s) 2014 published by polish botanical society fig. 3. leccinum variicolor: a – spores seen by lm, b – spores seen by sem. 74 m. stasińska and z. sotek © the author(s) 2014 published by polish botanical society ba-08 – pobrzeża południowobałtyckie littoral region: roby reserve; raised bog, vaccinio uliginosi-betuletum pubescentis, on peat and among sphagnum, under betula, 26.09.2008, leg. et det. m. stasińska, szub (stasińska 2011); myrico-salicetum auritae, on peat under betula, 19.09.2011, leg. z. sotek, det. m. stasińska, szub (unpublished). bb-01 – pobrzeża południowobałtyckie littoral regions: stramniczka reserve; raised bog, vaccinio uliginosi-betuletum pubescentis, on peat and among sphagnum, under betula, 27.09.2008, leg. et det. m. stasińska, szub (stasińska 2011); raised bog, vaccinio uliginosi-pinetum and erico-sphagnetum medii, among sphagnum, 20.09.2011, leg. z. sotek et m. stasińska, det. m. stasińska, szub (unpublished). bb-40 – pojezierza południowobałtyckie lakelands: near starea dobrzyca reserve; raised bog, vaccinio uliginosi-betuletum pubescentis, among sphagnum under betula, 18.10.2006, leg. et det. m. stasińska, szub (stasińska 2011). bc-07 – pojezierza południowobałtyckie lakelands: ostrzycki las-pierszczewko reserve; marginal part of peat bog, in mixed forest (e.g. fagus, alnus and betula), 06.09.2009, leg. m. wentoch-rekowska, det. m. wantoch-rekowski (unpublished, grej 2013)*. bg-10 – pojezierza wschdniobałtyckie lakelands: wigry national park; sphagno squarrosi-alnetum, on soil (halama, romański 2010)*. cb-91 – pojezierza południowobałtyckie lakelands: pszczew; peat bog, in mixed forest, among sphagnum, 20.09.2009, leg. m. wantoch-rekowski, det. t. ślusarczyk (unpublished, grej 2013)*. cc-74 – pojezierza południowobałtyckie lakelands: gościeszynek (1.7 km sw), n of shore of sykule duże lake; on soil, under betula, 11.10.2012, leg. et det. b. kudławiec (unpublished, grej 2013)*. cf-80 – niziny środkowopolskie lowlands: lasy łochowskie forests near łazy; vaccinio myrtilli-pinetum, under betula, 06.08.1985, leg. et det. z. domański; herbarium of z. domański, kram-f 52147 (unpublished). cg-55 – wysoczyzny podlasko-białoruskie high plains: białowieski national park; in forest, on soil, under betula, 06.10.1989, leg. a. m. słomczyńska and p. słomczyński, det. w. wojewoda, kram-f 30858 (unpublished). db-01 – pojezierza południowobałtyckie lakelands: rybojady reserve; betula pendula-betula pubescens community, on soil and among sphagnum, 10.07.2005, leg. et det. t. ślusarczyk (ślusarczyk 2007). dg-90 – polesie region: jezioro czarne sosnowickie reserve; vaccinio uliginosipinetum, among calluna under betula, 24.07.1984, leg. z. flisińska, lblm (flisińska 1987 (1988)). ec-28 – niziny środkowopolskie lowlands: siedliska (1.3 km sw), przedborów forest district, forest section no. 798; in wet mixed forest (e.g. alnus, betula and picea), on soil, 25.09.2012, leg. et det. p. zawada (unpublished, grej 2013)*. ec-29 – niziny środkowopolskie lowlands: łęki duże (0.6 km nw), przedborów forest district, forest section no. 795; in wet mixed forest (e.g. betula, carpinus and picea), on soil, 18.09.2012, leg. et det. p. zawada (unpublished, grej 2013)*. ec-37 – niziny środkowopolskie lowlands: leśniczówka koziołek (0.9 km ne), przedborów forest district, forest section no. 993; in wet mixed forest (e.g. alnus, betula, picea and pinus), on soil, 12.10.2012, leg. et det. p. zawada (unpublished, grej 2013)*. leccinum variicolor (basidiomycota, boletales) in poland 75 © the author(s) 2014 published by polish botanical society ed-02 – niziny środkowopolskie lowlands: korzeń reserve (złoczew forest district, forest section no. 359); transitional bog, on slope between eriophoro angustifolium-sphagnetum recurvi and leucobryo-pinetum, under betula, 30.09.2011, leg. et det. b. grzesiak, lod (unpublished). ed-13 – niziny środkowopolskie lowlands: uroczysko święte ługi (bełchatów forest district); eriophorum vaginatum-sphagnum fallax community, under betula, 19.09.2010, leg. et det. b. grzesiak, lod (unpublished). ed-47 – niziny środkowopolskie lowlands: kotków (1 km w); in humid broadleaved forest (e.g. alnus, betula and carpinus), on soil, 17.09.20011, leg. et det. j. nowicki (unpublished, grej 2013)*. ed-55 – wyżyna małopolska upland: (1) radziechowice i (2 km s); in broadleaved forest (e.g. alnus and betula), on soil; 02.09.2006, leg. et det. j. nowicki (unpublished, grej 2013)*; in scots pine and birch forest, on soil, 06.10.2011, leg. et det. j. nowicki, zbśril pan, 3/jn/20.12.11 (unpublished, grej 2013); (2) szczepocice prywatne (2 km e), in humid birch forest, on soil, 13.09.2009; leg. et det. j. nowicki, zbśril pan, 17/jn/15.10.09 (kujawa, gierczyk 2011); (3) łęg (0.75 km se); in wet scots pine and birch forest, on soil, 24.09.2011, leg. et det. j. nowicki (unpublished, grej 2013)*. ed-56 – wyżyna małopolska upland: leśniczówka strzałków (0.3 km s); in mixed forest (e.g. abies, and betula), on soil, 26.10.2008, 19.09.2010, leg. et det. j. nowicki, zbśril pan, 10/jn/10.12.10 (grej 2013*; kujawa, gierczyk 2013). ed-65 – wyżyna małopolska upland: jamrozowizna (1 km e); in mixed forest (e.g. pinus and betula), on soil, 10.09.2011, leg. et det. j. nowicki, zbśrilpan, 1/ jn/20.12.11 (unpublished, grej 2013). ed-67 – wyżyna małopolska upland: kotfin (1.7 km e); on the marginal part of peat bog in scots pine forest, under betula, on soil, 04.10.2009; leg. et det. j. nowicki, zbśrilpan, 16/jn/15.10.09 (kujawa, gierczyk 2011). ee-65 – wyżyna małopolska upland: czostkowa mt., klonów, abies alba-sphagnum girgensohnii community, among sphagnum under betula; 15.09.1999, leg. et det. j. łuszczyński, ktcb 1254 (łuszczyński 2007, 2008). eg-13 – polesie region: bachus reserve near chełm (sałata 1988; flisińska 2004)*. fd-22 – wyżyna śląsko-krakowska upland: świerklaniec (park); between forest and moist meadow, under betula pendula, tilia cordata and quercus robur, 14.09.1979, leg. et det. m. szczepka, kram-f 17997 (unpublished). fd-69 – wyżyna śląsko-krakowska upland: budzyń near mydlniczka, ca. 12 km ne of kraków; peat bog, 20.08.2006, leg. j. & m. piątek, det. m. piątek, kram-f 54998 (unpublished). fe-65 – północne podkarpacie region: łęki dolne, ca. 18 km e of tarnów, in humid birch forest, 13.08.1999, leg. anonymous, det. m. piątek, kram-f 39520 (unpublished). gg-61 – beskidy wschodnie mts: bieszczadzki national park, tarnawa reserve; raised bog, among sphagnum under betula, 18.08.2008, leg. et det. t. ślusarczyk; 18/ tś/bdpn/180808 (gierczyk et al. 2009). 76 m. stasińska and z. sotek © the author(s) 2014 published by polish botanical society discussion and conclusions in europe, leccinum variicolor is regarded as a rare species, more frequent only in northern europe (knudsen, taylor 2012; noordeloos, online). it usually occurs in different forest communities, in tree stands where birch, which is its mycorrhizal partner, is either a dominant species or an admixture (den bakker et al. 2004, 2007; legon et al. 2009). this would indicate that the species should be considerably more frequent. sometimes considerable morphological similarity is observed between leccinum variicolor and leccinum scabrum, and these two species may not be distinguished in the field. they are distinguished by the flesh at the stem base which turns blue-green in l. variicolor but the colouration may be visible after a few hours. both taxa also differ by habitat requirements. l. scabrum mainly grows in dry places (e.g. wojewoda 2003) while l. variicolor occurs in moist sites, wetlands and peatlands (chinan 2011; stasińska 2011; knudsen, taylor 2012). although it is widespread, leccinum variicolor is a rare and protected species in many european countries (e.g. siller et al. 2005) or it is red-listed, e.g., in romania (tănase, pob 2005; treated as vulnerable vu), in the czech republic (holec, beran 2006; treated as near threatened nt) and in hungary (siller, vasas 1995; treated as endangered, vulnerable and rare – category 2). in poland, the species is included on regional red lists of fungi only in upper silesia (category i – indeterminate, wojewoda 1999) and the góry świętokrzyskie mts (category r – rare, łuszczyński 2002; category nt – near threatened, łuszczyński 2008). it may be recommended to include the species on the national red list of macrofungi (wojewoda, ławrynowicz 2006) as nearly half of all its records in poland to date are from peatlands and its margins (cf. the list of localities). according to krieglsteiner (2000), wetland draining, occurring worldwide for many years, is a threat to l. variicolor. the distribution of l. variicolor in poland is not recognized well and the number of its sites may be greater than it is believed. based on the available published and unpublished data and our observations, the species is probably rare and threatened in poland. acknowledgements. the authors would like to thank the curators of all the herbaria for the loan of specimens. we are grateful to dr. barbara grzesiak (university of łódź) for making her unpublished material available for publication in the present paper. we would also like to express our gratitude to dr. bożena białecka and dr. magdalena bihun (centre for molecular biology and biotechnology, environmental testing laboratory, university of szczecin (poland)) for taking sem pictures. we are indebted to the anonymous reviewer for helpful suggestions on the manuscript. references chinan v. 2011. macrofungi from „grădiniţa“ peat bog (eastern carpatians, romania). biologie vegetală 57 (1): 35-40. den bakker h.c., noordeloos m.e. 2005. a revision of european species of leccinum gray and notes on extralimital species. persoonia 18: 511-587. den bakker h.c., zuccarello g.c., kuyper th.w., noordeloos m.e. 2004. evolution and host specificity in the ectomycorrhizal genus leccinum. new phytologist 163: 201-215. leccinum variicolor (basidiomycota, boletales) in poland 77 © the author(s) 2014 published by polish botanical society den bakker h.c., zuccarell, g.c., kuyper th.w., noordeloos m.e. 2007. phylogeographic patterns in leccinum sect. scabra and the status of the arctic-alpine species l. rotundifoliae. mycological research 3: 663-672. flisińska z. 1987 (1988). macromycetes zbiorowisk leśnych i torfowiskowych pojezierza łęczyńsko włodawskiego. acta mycol. 23 (1): 19-92. flisińska z. 2004. grzyby lubelszczyzny. i, ii. lubelskie tow. nauk., merpol s.c., lublin. fu s.-z., wang q.-b., yao y.-j. 2006. an annotated checklist of leccinum in china. mycotaxon 96: 47-50. gierczyk b., chachuła p., karasiński d., kujawa a., kujawa k., pachlewski t., snowarski m., szczepkowski a., ślusarczyk t., wójtowski m. 2009. grzyby wielkoowocnikowe polskich bieszczadów. i. parki nar.: rez. przyr. 28 (3): 3-100. grej 2013. leccinum variicolor watling. grej – rejestr gatunków grzybów chronionych i zagrożonych w polsce. http://www.grzyby.pl/gatunki/leccinum_variicolor.htm halama m., romański m. 2010. grzyby makroskopijne (macromycetes). 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(ed.). 2000. atlas of the geographical distribution of fungi in poland. 1. w. szafer institute of botany, polish academy of sciences, kraków. wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków. wojewoda w., ławrynowicz m. 2006. red list of the macrofungi in poland. (in:) z. mirek, k. zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. 3 rd. ed. w. szafer institute of botany, polish academy of sciences, kraków: 53-70. leccinum variicolor (basidiomycota, boletales) w polsce streszczenie leccinum variicolor watling (koźlarz różnobarwny), jest gatunkiem szeroko rozprzestrzenionym, prawdopodobnie o zasięgu cyrkumborealnym. najczęściej zajmuje wilgotne siedliska, rośnie w różnego typu lasach oraz w miejscach otwartych i na torfowiskach. w pracy zamieszczono szczegółowy opis i ilustracje najważniejszych cech makroi mikromorfologicznych owocników koźlarza różnobarwnego. przedstawiono również informacje na temat taksonomii, ekologii i rozmieszczenia na świecie, oraz aktualną mapę jego rozmieszczenia w polsce. autorzy podkreślają, że na obszarze polski jest to gatunek rzadki i prawdopodobnie zagrożony oraz proponują ujęcie tego taksonu na czerwonej liście grzybów wielkoowocnikowych w polsce. 2014-06-30t22:21:55+0200 polish botanical society 2014-08-25t21:38:57+0200 polish botanical society 2014-08-29t18:50:08+0200 polish botanical society 2014-08-25t21:39:25+0200 polish botanical society 2014-08-28t20:05:15+0200 polish botanical society 2014-08-31t21:53:51+0200 polish botanical society 2014-08-25t14:56:26+0200 polish botanical society 2014-08-20t22:12:58+0200 polish botanical society 2014-08-20t22:53:53+0200 polish botanical society 2014-08-26t20:31:45+0200 polish botanical society 2014-08-31t21:56:39+0200 polish botanical society 2014-08-31t21:53:48+0200 polish botanical society 2014-08-28t14:43:43+0200 polish botanical society 2014-08-31t21:54:51+0200 polish botanical society 2014-08-31t21:55:39+0200 polish botanical society 2014-08-27t16:49:35+0200 polish botanical society 2014-08-29t18:50:43+0200 polish botanical society 2014-09-06t20:24:11+0200 polish botanical society 2014-08-28t17:48:15+0200 polish botanical society 2014-08-27t16:35:58+0200 polish botanical society 2014-08-31t21:55:42+0200 polish botanical society 2014-08-27t17:37:58+0200 polish botanical society three species of the genus agaricus new to poland janusz łuszczyński department of botany, institute of biology, jan kochanowski university świętokrzyska 15, pl-25-406 kielce, jluszcz@ujk.kielce.pl łuszczyński j.: three species of the genus agaricus new to poland. acta mycol. 43 (1): 161–165, 2008. the paper presents informations on agaricus excellens (f. h. møller) f. h. møller, a. maleolens f. h. møller and a. romagnesii wasser, three new species for poland. key words: agaricus excellens, a. maleolens, a. romagnesii, góry świętokrzyskie mts., kielce, poland introduction fruitbodies of fungi belonging to the genus agaricus are characterised by a white or yellow pileus and free gills with a regular trama when young, later becoming irregular. spores are globose, oval or elliptical and the spore print is purple-brown to black-brown. the spore wall is smooth, often thick and indistinctly pseudoamyloid. cheilocystidia either occur or are absent. the shape, size and attachment of the ring are important diagnostic features of the genus. much attention is also paid to the reaction of the flesh in aniline, concentrated hno3, strong naoh or koh and to the discolouration of the cuticle or the flesh on exposure to air when the fruitbody is rubbed or broken. despite large fruitbodies, few fungi of the genus agaricus are known in poland and were until recently represented merely by 28 species (wojewoda 2003). in contrast, ca. 55 species are known in germany (krieglsteiner 1991), poland’s neighbour where climatic and habitat conditions are similar. nearly all species of agaricus known to grow in europe occur there (horak 2005). in poland, only four species: a. arvensis, a. campestris, a. silvaticus and a. silvicola, can be regarded as common, and further six: a. augustus, a. bitorquis, a. comptulus, a. niveolutescens, a. semotus and a. xanthodermus as quite frequent with the number of known localities ranging from 10 and 20. the other 18 species are known only from few or single localities. often cosmopolitan, species of this genus occur in a variety of habitats: natural (forests), seminatural non-forest environments (meadows, shrubs and pastures), acta mycologica vol. 43 (2): 161–165 2008 162 j. łuszczyński as well as segetal (fields, gardens) and ruderal (parks, lawns); usually, however, in sites rich in humus. three species of the genus agaricus, a. excellens, a. maleolens and a. romagnesii, new to the polish mycobiota were found during mycocoenobiotic studies in the góry świętokrzyskie mts. (łuszczyński 2007, 2008). the collected material was deposited in the herbarium of the department of botany, institute of biology, jan kochanowski university, kielce. species description agaricus excellens (f. h. møller) f. h. møller, friesia 4: 204 (1952). syn.: psalliota augusta fries sensu ricken, die blätterpilze, p. 235 (1915). – agaricus urinascens (jul. schäff. & f.h. møller) singer, lilloa 22: 431 (1951). – psalliota excellens f.h. møller, friesia 4: 178 (1952). – agaricus macrosporus subsp. excellens (f.h. møller) bohus, ann. hist.-nat. mus. natl. hung. 70: 105 (1978). – agaricus macrosporus var. excellens (f.h. møller) vasas, ann. hist.-nat. mus. natl. hung. 82: 41 (1990). – agaricus urinascens var. excellens (f.h. møller) nauta, persoonia 17(3): 462 (2001) [2000]. iconography: møller 1952. friesia 4, fig. 29, pl. xxxiii. – essette. 1964. les psalliotes, tab. 44. – wasser 1980. flora fungorum rss ucrainicae, figs. 68-69. – phillips 1981. mushrooms, p. 167. – cappelli 1984. agaricus l.: fr. ss. karsten (psalliota fr.), text fig. 29 + icon. fig. 41. – michael et al. 1985. handbuch für pilzfreunde 4, tab. 15. – wasser 1985. agarikovyje griby, tab. 4(6). – hagara 1987. atlas húb, p. 324. description. species of the section flavescentes. pileus spherical convex, at first pure white, finely squamulose along the margin in young fruitbodies. cuticle discolours yellow, chrome-yellow when rubbed. pileus ochre-yellow in older fruitbodies, up to 15 cm. in diam, densely covered in fine squamules. stipe white, clavate, up to 12 cm long, 3 cm in diam., squamulose in lower part, glabrous in upper part. ring strong, pendant, squamulose on the outside. flesh white, becoming pink on cutting, smell slightly amygdaline. spores 9.45-10.8 × 5.4-6.7 �m. chei�m. cheim. cheilocystidia numerous, oval, clavate, 15-30 × 5.4-14.8 �m (fig. 1). localities. in forests: peucedanopinetum and tilio-carpinetum, on ground, september-october, localifig. 1. agaricus excellens: a – spores, b – cystidia. fig. 2. localities of agaricus excellens, a. maleolens and a. romagnesii in square of the atpol net. three species of the genus agaricus 163 ties: kielce (mt. biesak, bukówka, mt. telegraf) and dyminy. all localities in the atpol square ee 74 (fig. 2). distribution. the fungus occurs in many european countries, for instance in austria, belgium, bulgaria, the czech republic, denmark, france, germany, great britain, hungary, italy, the netherlands, norway, portugal, slovakia, slovenia, switzerland, ukraine (bohus 1978; hansen, knudsen 1992; hernández-crespo 2006; kreisel 1987; krieglsteiner 1991; lacheva 2006; wasser 1980, 1985). it is also known from eastern asia (krasnoyarsk krai, wasser 1985) and africa from morocco (malençon, bertault 1970). agaricus maleolens f. h. møller, friesia 4: 203 (1952). syn.: agaricus campestris subsp. bernardii (quél.) konrad & maubl., icones 6 (texte): 60 (1937). – agaricus ingratus (f.h. møller) pilát, acta musei nationalis pragae, 7 b, 1 (1951). (nom. inval.). – pratella bernardii (quél.) quél., fl. mycol. france (paris): 73 (1888). – psalliota bernardii (quél.) quél. [as ‘bernardi’], bull. soc. bot. france 25: 288 (1879) [1878]. – psalliota ingrata f.h. møller, friesia 4: 17 (1950), non agaricus ingratus fries (= gymnopus confluens). iconography. møller 1952. friesia 4, fig. 5, pl. viii. – wasser 1980. flora fungorum rss ucraini-friesia 4, fig. 5, pl. viii. – wasser 1980. flora fungorum rss ucrainicae, fig. 62, tab. ix. – michael et al. 1985. handbuch für pilzfreunde 4, tab. 9. – cappelli 1984. agaricus l.: fr. ss. karsten (psalliota fr.), text fig. 3 + icon. fig. 3. – wasser 1985. agarikovyje griby, tab. 4(1). description. pileus hemispherical convex, glabrous, 5-7 cm in diam., white-ochre. margin draped with the remains of the partial veil. gills thin, crowded, chocolatecoloured. stipe cylindrical, evenly thick, concolourous with pileus, 6-8 x 1.8-2 cm. ring thin, spreading, falls down pulling away from the stipe. flesh ochrepink on cutting, darkening, darkening particularly quickly in the stipe and becoming brunneous-ochre with a distinctive pink tint. smell of fresh fish, stronger and unpleasant fishy smell when drying. spores broadly elliptical, 5.4-6.6 x 4.2-5 �m, redbrown, mostly with a drop of fat (fig. 3). cheilocystidia hyaline, cylindrical, with a blunt hemispherical tip, erect or arching ascendant, 6-8 �m in diam., on gill edges. locality. in urban area, on lawn, under populus sp., may, locality: kielce (szydłówek), atpol square ee 74. distribution. rare throughout europe; known from germany, switzerland (krieglsteiner 1991), denmark, finland, sweden (hansen, knudsen 1992), bulgaria (lacheva 2006), hungary, slovakia, the czech republic, spain, portugal, italy, the netherlands, france, ukraine (wasser 1985). also known from africa from morocco (malençon, bertault 1970). agaricus romagnesii wasser, ukrainian botanical journal 34: 305 (1977). syn.: agaricus radicatus (vittad.) romagnesi, bull. soc. mycol. france, 53, p. 129 (1937) (nom. inval.), non agaricus radicatus rehl.: fries 1821. – psalliota radicata (vittad.) essette, les psalliotes, tab. 22 (1964) (nom. inval.). – agaricus bresadolanus bohus, ann. hist.-nat. mus. natl. hung. 61: 154 (1969). – agaricus bresadolanus bohus sensu reid, fungorum rariorum icones coloratae 6, p. 6 (1972). – psalliota infida alessio, micol. ital. 4(2): 21 (1975) (nom. inval.). – agaricus infidus (alessio) bon, doc. mycol. 11(44): 28 (1981) (nom. inval.), non agaricus infidus peck 1900. iconography. romagnesii 1937. bull. soc. mycol. france, 53, p. 130. – essette 1964. les psalliotes, tab. 22. – bohus 1971. ann. hist.-nat. mus. natl. hung. 63, p. 79. – reid 1972. fungorum rariorum fig. 3. agaricus maleolens: a – spores, b – cystidia. 164 j. łuszczyński icones coloratae 6, pl. 42. – rinaldi et al. 1974. l’atlante dei funghi, fig. 4. – alessio 1975. micol. ital. 4(2), tab. 11. – wasser 1977. ukrainian botanical journal 34(3): 85. – wasser 1980. flora fungorum rss ucrainicae, tab. i(1). – blatto 1982. atlante fotografico dei funghi, tab. 31. – cappelli 1984. agaricus l.: fr. ss. karsten (psalliota fr.), text fig. 40 + icon. fig. 70. – wasser 1985. agarikovyje griby, tab. 1 (3). description. pileus at first white, later darker, beige, covered in light-brown, brown squamules and fibres; spherically convex when young, flattened, centrally depressed, up to ca. 8 cm in diam. in older specimens. stipe white, later rusty-yellowing, cylindrical, slightly thickened at the base, contracting into quite thick rhizomorphic strands, one or more. ring thin, delicate, not persistent. gill edges sterile, with quite numerous hyaline, clavate cheilocystidia, 30-40 x 8.1-10.8 �m. spores broadly elliptical, 6.5-8 x 3.5-5 �m (fig. 4). locality. in pastures: lolio-cynosuretum, on ground, july, locality: kielce (nowy folwark), atpol square ee 74. distribution. in southern and central europe; known from slovenia, germany, france, bulgaria, the czech republic, slovakia, ukraine, hungary, great britain, italy, spain, portugal (hernández-crespo 2006; lacheva 2006; parra 2003). also known from asia, israel, uzbekistan, turkmenistan, tadzhikistan (wasser 1985), turkey (kaşik et al. 2003), georgia [http://www.cybertruffle.org.uk/ gruzmaps/a/0020960_.htm]. references alessio c. l. 1975. psalliota radicata vitt. sensu bres. e p. radicata sensu essette: nome nuovo per la seconda: psalliota infida (= agaricus infidus) sp.n. micol. ital. 4(2): 16–22. blatto l. 1982. atlante fotografico dei funghi. hoepli. milano. bohus g. 1971. agaricus studies. iii. ann. hist.-nat. mus. natl. hung. 70: 105–110. bohus g. 1978. agaricus studies. viii. ann. hist.-nat. mus. natl. hung. 63: 77–82. bohus g. 1990. agaricus studies. xi (basidiomycetes, agaricaceae). monographical key. ann. hist.-nat. mus. natl. hung. 82: 39–59. bon m. 1981. novitates. combinaisons et noms nouveaux. doc. mycol. 11 (44): 1–28. cappelli a. 1984. agaricus l.: fr. ss. karsten (psalliota fr.). fungi europaei. libreria editrice biella giovanna. saronno. essette h. 1964. les psalliotes. lechevalier. paris. hagara l. 1987. atlas húb. vydavateľstvo osveta. martin. hansen l., knudsen h. 1992. (eds). 1992. nordic macromycetes. 2. polyporales, boletales, agaricales, russulales. nordsvamp. copenhagen. hernández-crespo j. c. 2006. s i m i l. an on line information system of the iberian fungal diversity. royal botanic garden madrid, c.s.i.c. research project flora micológica ibérica i-vi (1990-2008). spanish ministry of education and science. [http://www.rjb.csic.es/fmi/sim.php]. horak e. 2005. röhrlinge und blätterpilze in europa. elsevier, spektrum akademischer verl. münchen. kaşik g., türkoğlu a., doğan h. h., őstürk c. 2006. macrofungi of yahjalı (kayseri) province. turkish journal of botany 27: 453–462. konrad p., maublanc a. 1937. icones selectae fungorum, 6. lechevalier. paris. fig. 4. agaricus romagnesii: spores. three species of the genus agaricus 165 kreisel h. (ed.). 1987. pilzflora der deutschen demokratischen republik. basidiomycetes (gallert-, hut und bauchpilze). veb g. fischer verl. jena. krieglsteiner g. j. 1991. verbreitungsatlas der großpilze deutschlands (west). 1. teil b. verl. eugen ulmer. stuttgart. lacheva m. n. 2006. genus agaricus l.: fr. emend. p. karst. (mushroom) in bulgaria – taxonomy, ecology, chorology and economical importance. ph. d. thesis. agrar. univ. plovdiv [http://botanica.hit. bg/doc/agaricusphdthesis.pdf]. łuszczyński j. 2007. diversity of basidiomycetes in various ecosystems in góry świętokrzyskie mts. monogr. bot. 97: 1–218. łuszczyński j. 2008. basidiomycetes of the góry świętokrzyskie mts. a checklist. wyd. uniwersytetu humanistyczno-przyrodniczego jana kochanowskiego w kielcach, kielce (in print). malençon g., bertault r. 1970. flore des champignons superieurs du maroc. 1. faculte des science. rabat. michael e., hennig b., kreisel h. 1985. handbuch für pilzfreunde, 4. veb g. fischer verl. jena. møller f. h. 1952. danish psalliota species, i-ii. friesia 4: 135–220. nauta m. m. 2001 [2000]. notulae ad floram agaricinam neerlandicam. xxxvii. notes on agaricus section arvenses. persoonia 17 (3): 457-463. parra l. a. 2003. números 2070-2123. (in:) j. c. hernández (ed.). bases corológicas de flora micológica ibérica. números 2070-2178 caudernos de trabajo de flora micológica ibérica 19: 19–104. pilát a. 1951. the bohemian species of the genus agaricus. acta musei nationalis pragae, 7 b, 1: 1–142. phillips r. 1981. mushrooms. pan books ltd, londra. quélet l. 1878. quelques espèces nouvelles de champignons (6. suppl.). bull. soc. bot. france 25: 288. quélet l. 1888. flore mycolgique de la france et des pays limitrophes. octave doin, paris. reid d. a. 1972. fungorum rariorum icones coloratae, 6. j. cramer, vaduz. ricken a. 1915. die blätterpilze. leipzig. rinaldi a., tyndalo v., pace g. 1974. l’atlante dei funghi. mondadori. romagnesi h. 1937. liste des champignons supérieurs recueillis à paris. bull. soc. mycol. france 53: 128–131. singer r. 1951. the “agaricales” (mushrooms) in modern taxonomy, lilloa 22: 1–832. wasser s. p. 1977. new and rare species of agaricaceae cohn. family. ukrainian botanical journal 34 (3): 305–307. wasser s. p. 1980. agaricaceae cohn. flora fungorum rss ucrainicae. naukova dumka.kiev. wasser s. p. 1985. agarikovyje griby sssr. an ussr. naukova dumka. kiev. wojewoda w. 2003. checklist of polish larger basidiomycetes. (in:) z. mirek (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków. gatunki z rodzaju agaricus nowe dla polski s t r e s z c z e n i e rodzaj agaricus reprezentowany jest w polsce przez 28 gatunków. autor przedstawia trzy nowe dla polski gatunki z tego rodzaju: a. excellens, a. maleolens i a. romagnesii zebrane podczas długoterminowych badań mikocenologicznych w górach świętokrzyskich. agaricus excellens został odnaleziony w zbiorowisku peucedano-pinetum i tilio-carpinetum na ziemi, a. maleolens na ziemi pod populus sp. w strefie miejskiej kielc, a a. romagnesii w zbiorowisku lolio-cynosuretum. autor podaje szczegółowe opisy morfologiczne owocników, siedlisk występowania i zbioru oraz rozmieszczenie geograficzne w polsce i europie. 2014-01-01t11:47:59+0100 polish botanical society 2014-08-28t17:49:02+0200 polish botanical society 2014-08-31t21:55:07+0200 polish botanical society 2014-08-25t19:07:36+0200 polish botanical society 2014-09-06t20:24:14+0200 polish botanical society 2014-08-29t18:50:16+0200 polish botanical society 2014-09-06t20:24:30+0200 polish botanical society 2014-08-28t20:05:26+0200 polish botanical society 2014-08-31t21:54:27+0200 polish botanical society 2014-08-31t21:57:39+0200 polish botanical society 2014-08-31t21:57:31+0200 polish botanical society 2014-08-31t21:56:36+0200 polish botanical society 2014-08-31t21:55:16+0200 polish botanical society 2014-09-02t20:09:15+0200 polish botanical society 2014-08-26t20:32:11+0200 polish botanical society 2014-09-06t20:24:47+0200 polish botanical society lichens of red oak quercus rubra in the forest environment in the olsztyn lake district (ne poland) dariusz kubiak department of mycology, university of warmia and mazury in olsztyn oczapowskiego 1a, pl 10 957 olsztyn, darkub@uwm.edu.pl k u b i a k d.: lichens of red oak quercus rubra in the forest environment in the olsztyn lake district (ne poland). acta mycol. 41 (2): 319 328, 2006. a list of 63 species of lichens and 4 species of lichenicolous fungi recorded on the bark of red oak (quercus rubra l.) in poland is given. literature data and the results of field studies conducted in the forest in the olsztyn lake district between 1999 and 2005 are used in the report. fifty five taxa, including lichens rare in poland, for instance lecanora albella, lecidella subviridis, ochrolechia turneri, were recorded. key words: lichens (lichenized fungi), lichenicolous fungi, red oak (quercus rubra), poland introduction rich lichen biotas, usually comprising numerous specific species, are associated with the genus quercus l. both in poland (c i e ś l i ń s k i , to b o l e w s k i 1988; r u t k o w s k i 1995) and in other parts of europe (a l v a r e z , c a r b a l l a l 2000; z e d d a 2002; e n g e l et al. 2003). many rare or very rare lichen species in poland and taxa considered to be extinct have been found to colonise oak bark in poland (c i e ś l i ń s k i , to b o l e w s k i 1988; r u t k o w s k i 1995; r u t k o w s k i , k u k w a 2000; fa ł t y n o w i c z 1991). its diversified texture provides niches suitable for different lichen species while the phorophyte’s longevity influences the richness and diversity of its lichen biota. the occurrence of native oak species (quercus robur, q. petraea) in various forest communities is also significant (m a t u s z k i e w i c z 2001; m o d r z y ń s k i et al. 2006; d a n i e l e w i c z , p a w l a c z y k 2006). however, despite abundant data on the occurrence of lichens on oak bark, the lichen biota of this phorophyte has not been studied thoroughly and extensively. in poland, the genus quercus is represented by three native species (q. robur, q. petraea, q. pubescens) and several alien, mostly north american, species. quercus rubra l. is the only taxon of many introduced species to have acclimatised in natural forest habitats in poland (k r ó l 1967; h e r e ź n i a k 1992). at the same time, the number of introduction sites and its total cultivation area exceed those of all other acta mycologica vol. 41 (2): 319-328 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 320 d. kubiak deciduous species introduced in polish forests (d a n i e l e w i c z , p a w l a c z y k 2006). quercus rubra is native to eastern parts of north america. it usually grows in lowland and submontane areas, and reaches 1600 m above sea level in the mountains (b r o w i c z 1953). its native range covers the area from nova scotia in the north to alabama and texas in the south, reaching the belt of central american prairies in the west, and it generally corresponds to the eastern (appalachian) forest region where deciduous forests that lose their leaves for winter are the basic type of vegetation (appalachian oak forest region) (p o d b i e l k o w s k i 1995). in the native range, quercus rubra grows on a variety of soils and in topographically diversified areas, often forming pure stands. species of the genus quercus, both native and acclimatised, have usually been treated inclusively in lichenological research in poland. although some authors draw attention to the interesting lichen biota of red oak (z a l e w s k a et al. 2004), only few provide extensive relevant information (ty s z k i e w i c z 1935; d z i a b a s z e w s k i 1962; g l a n c 1969; g l a n c , k a p u ś c i ń s k i , k r ó l 1971; k u k w a 2002; k u b i a k 2005). the lack of detailed studies on species diversity, particularly important in the case of q. rubra, the most widespread of alien species, encouraged the present author to examine the lichen biota of this phorophyte in depth. study area quercus rubra sites located in the olsztyn lake district in the municipal forests of the city of olsztyn, in the las warmiński reserve and the kamienna góra reserve as well as in forest sections 51, 104 and 523 of the nowe ramuki forest district were examined. the sites are usually mid-forest cultivations covering a total surface area of 0.10-0.05 ha. the tree forms avenues, and occurs along section lines or forest roads less frequently. single trees, probably self-sown, are also noted. the oldest sites of quercus rubra in the area date back to the 1880s and are mostly the remains of experimental forest areas established by german foresters (cf. tu m i ł o w i c z 1965, 1967). the majority of the study sites come from this period. the olsztyn lake district belongs to the western part of the masurian lake district (k o n d r a c k i 1998). the region is characterised by significant terrain diversification. rolling uplands of the clayey ground moraine constitute the northern part of the olsztyn lake district, while hilly uplands and kettles in outwash plains, built from sands and gravels covered by expansive forests called the nidzica forest (z a r ę b a 1978), abundant in lakes, form the southern part. the altitudes of moraine hills along the peripheries of the mesoregion slightly exceed 200 m above sea level and are significantly lower in the central part. a greatly diversified plant cover is a feature of the olsztyn lake district, located in the transitional zone where elements of the oceanic and continental climates clash. occurrence ranges of numerous plants, including forest-forming trees, characteristic both of central and western europe (atlantic flora) and of northern europe (subboreal and boreal species), run and cross in the area. durmast oak quercus petraea (b o r a t y ń s k a 1979), sycamore acer pseudoplatanus (b o r a t y ń s k i 1979) and beech fagus sylvatica (to k a r z 1971) reach their north-eastern limits of occurrence ranges in the olsztyn lake district. norway spruce picea abies (j u t r z e n k a -tr z e b i a t o w s k i , f e n y k 2001) reaches the south-western limit of the boreal lichens of red oak 321 range. the ranges of main forest-forming species determine the shape and character of entire forest communities. associations characterised by the continental type of range: peucedano-pinetum, seratulo-pinetum and tilio-carpinetum (subboreal variety), reach their western limits in the area while associations characterised by the subatlantic range or similar to it: fago-quercetum, luzulo pilosae-fagetum, stellariocarpinetum, galio odorati-fagetum or leucobryo-pinetum, reach their eastern limits (m a t u s z k i e w i c z 2001). material and methods detailed field studies were conducted in 2005. data collected by the author in the period between 1999 and 2005 were additionally used. in total, 16 querqus rubra sites located within forest complexes previously studied lichenologically were analysed. critical taxa (sterile sorediate lichens, species of the genus usnea) were determined using chromatography methods (tlc), performed according to the procedures specified by o r a n g e , j a m e s and w h i t e (2001), in the department of mycology, university of warmia and mazury in olsztyn, and the department of plant taxonomy and nature protection, university of gdańsk. names of lichens and lichnicolous fungi are given after fa ł t y n o w i c z (2003) and k u k w a and d i e d e r i c h (2005), red list categories of lichens after c i e ś l i ń s k i , c z y ż e w s k a and fa b i s z e w s k i (2003) and c i e ś l i ń s k i (2003b). the collected herbarium material is deposited in the herbarium of the department of mycology, university of warmia and mazury in olsztyn (oltc-l). the taxa are listed alphabetically. source studies are specified in the case of taxa reported in earlier publications. the number of sites and their general location is given for the taxa recorded in the present study (tab. 1). results a total of 55 species of lichens and 4 species of lichenicolous fungi were recorded in the study sites (tab. 1). between 11 and 25 species were recorded in individual sites, and 19 species on average. taxa with crustose thalli dominate (39 species); lichens with foliose thalli (8 species) or with fruticose thalii (5 species) and dimorphic species (cladonia spp.; 4 species) are less numerous. the following species were recorded most frequently: hypogymnia physodes, cladonia coniocraea, platismatia glauca, lepraria incana, mycoblastus fucatus, pertusaria amara, parmelia sulcata, phlyctis argena, buellia griseovirens, lecanora pulicaris, melanelia fuliginosa, parmeliopsis ambiqua and ochrolechia androgyna. except ochrolechia androgyna, these thalli are frequent or common, moderately or significantly ubiquitous. this group is responsible for the formation of crustose or crusote-foliose epiphytic communities, typical of this phorophyte, where taxa with fruticose thalli are recorded only sporadically. twelve taxa (23.2% of the biota) were recorded in single sites, mostly on single trees. these are: amandinea punctata, anisomerydium polyporii, arthonia spadicea, bacidina arnoldiana, chaenotheca ferruginea, chaenothecopsis pusilla, lecanora albella, l. carpinea, lecidella subvirdis, ochrolechia turneri, placynthiella icmalea, scoliciosporum chlorococcum and xanthoria polycarpa. both common species, characterised by a broad ecological scale, and rare or very rare species 322 d. kubiak ta b l e 1 lichens and lichenicolous fungi recorded on quercus rubra in poland species literature data olsztyn lake district number of sites site location lichenized fungi (lichens) amandinea punctata (hoffm.) coppins et scheid. 1 nr anisomerydium polypori (m.b. ellis et everh.) m.e. barr 1 ol arthonia radiata (pers.) ach. ty s z k i e w i c z (1935), as a. vulgaris schaer f. astroidea ach. arthonia spadicea leight. 1 ol bacidina arnoldiana (körb.) v. wirth et vězda 1 ol bacidina phacodes (körb.) vězda k u b i a k (2005) 4 nr, ol biatora efflorescens (hedl.) erichsen 2 ol buellia griseovirens (turner et borrer ex sm.) almb. 11 lw, nr, ol chaenotheca chrysocephala (ach.) th. fr. 4 ol chaenotheca ferruginea (turner ex sm.) mig. 1 ol cladonia coniocraea auct. 16 kg, lw, nr, ol cladonia digitata (l.) hoffm. 1 ol cladonia fimbriata (l.) fr. 6 kg, lw, ol dimerella pineti (ach.) vězda 9 lw, ol evernia prunastri (l.) ach. ty s z k i e w i c z (1935), g l a n c (1969) 9 lw, nr, ol fuscidea arboricola coppins et tønsberg 8 nr c.ap., ol fuscidea pusilla tønsberg 2 ol graphis scripta (l.) ach. ty s z k i e w i c z (1935) 4 lw, ol hypocenomyce scalaris (ach.) choisy 9 kg, lw, nr, ol hypogymnia physodes (l.) nyl. ty s z k i e w i c z (1935), as parmelia physodes f. labrosa (l.) ach.; kukwa (2003) 16 kg, lw, nr, ol lecanora albella (pers.) ach. ty s z k i e w i c z (1935), as l. albella (pers.) ach. var. subalbella nyl., k u b i a k (2005) 1 ol lecanora argentata (ach.) malme 1 lw lecanora carpinea (l.) vain. 1 lw lecanora conizaeoides nyl. ex crombie g l a n c et al. (1971), as l. conizea (ach.) nyl. 3 kg, ol lecanora expallens ach. k u k w a (2002) 3 nr, ol lecanora pulcaris (pers.) ach g l a n c et al. (1971), as l. chlarona 10 kg, lw, ol lecidella subviridis tønsberg 1 ol lecidella eleaochroma (ach.) choisy ty s z k i e w i c z (1935), as lecidea parasema ach. lepraria elobata tønsberg 5 lw, ol lepraria incana (l.) ach. 13 lw, nr, ol lepraria jackii tønsberg 3 kg, ol lepraria lobificans nyl. 4 ol lepraria rigidula (b. de lesd.) tønsberg 4 kg, lw, nr, ol melanelia fuliginosa (fr. ex duby) essl. ty s z k i e w i c z (1935), as parmelia fuliginosa (e.fr.) nl.; d z i a b a s z e w s k i (1962), as parmelia fuliginosa (fr.) nyl. 11 lw, kg, ol lichens of red oak 323 melanelia olivacea (l.) essl. ty s z k i e w i c z (1935), as parmelia olivacea (l.) nyl. melanelia subarifera (nyl.) essl. g l a n c (1969), as parmelia subarifera nyl. micarea prasina fr. g l a n c (1969), as catillaria prasina (fr.) th.fr.; k u k w a (2002) 9 lw, nr, ol mycoblastus fucatus (stirt.) zahlbr. 14 kg, lw, nr, ol ochrolechia androgyna (hoffm.) arnold 10 nr, lw, ol ochrolechia microstictoides räs. 3 nr, ol ochrolechia turneri (sm.) hasselrot 1 lw parmelia saxatilis (l.) ach. 3 lw, ol parmelia sulcata taylor ty s z k i e w i c z (1935), k u k w a (2002) 12 lw c.ap., ol parmeliopsis ambigua (wulfen ex jacq.) nyl. d z i a b a s z e w s k i (1962), k u k w a (2002) 1 ol pertusaria amara (ach.) nyl. d z i a b a s z e w s k i (1962), k u k w a (2002) 14 lw, nr, ol pertusaria coccodes (ach.) nyl. 7 kg, lw, nr, ol pertusaria pertusa (weigel.) tuck. var. pertusa ty s z k i e w i c z (1935), as p. communis d.c. phlyctis argena (ach.) flot. k u k w a (2002) 12 lw, nr, ol physcia adscendens (fr.) h. olivier ty s z k i e w i c z (1935) physconia distorta (with.) j.r. laundon ty s z k i e w i c z (1935), as physcia pulverulenta (hoffm.) nyl. placynthiella icmalea (ach.) coppins et p. james 1 kg platismatia glauca (l.) w.l. culb. et c.f. culb. k u k w a (2002) 15 lw, nr, ol pseudevernia furfuracea (l.) zopf. 3 kg, lw, nr pseudosageda aenea (wallr.) hafellner et kalb 2 ol ramalina farinacea (l.) ach. ty s z k i e w i c z (1935) 2 ol ramalina fraxinea (l.) ach. ty s z k i e w i c z (1935) ramalina pollinaria (westr.) ach. ty s z k i e w i c z (1935), g l a n c (1969) ropalospora viridis (tønsberg) tønsberg 7 lw, ol scoliciosporum chlorococcum (graeve ex stenh.) vězda g l a n c (1969), as bacidia chlorococca (graeve) lett. 1 kg usnea filipendula stirt. 2 lw, ol usnea subfloridana stirt. 3 lw, ol vulpicidia pinastri (scop.) j. e. mattson et m.j. lai 2 lw, ol xanthoria polycarpa (hoffm.) rieber 1 kg lichenicolous fungi chaenothecopsis pusilla (ach.) a. f.w. schmidt (on wood) 1 ol clypeococcum hypocenomycis d. hawksw. (on hypocenomyce scalaris) 1 kg lichenoconium erodens m.s. christ. et d. hawksw. (on lecanora conizaeoides and hypogymnia physodes) 1 kg monodictys epilepraria kukwa et diederich (on lepraria sp.) k u k w a (2004) 1 ol abbreviations: kg kamienna góra reserve; lw las warmiński reserve; ol olsztyn city forests; nr nowe ramuki forest district; c. ap. cum apotheciae 324 d. kubiak whose ecological preferences are poorly known belong to this group. one of the first records of lecidella subviridis in lowland poland was also identified (cf. c z a r n o t a , k u k w a 2003; k u k w a 2006). lichens were observed not only on the bark of tree trunks but also on exposed dead trunk wood and branches fallen on the ground. the following taxa were recorded on dead wood: anisomerydium polyporii, chaenotheca chrysocephala, chaenothecopsis pusilla, hypocenomyce scalaris, lepraria incana and cladonia sp. it was the only occurrence substrate of two species: anisomerydium polyporii and chaenothecopsis pusilla. the following species occurred on fallen branches: hypogymnia physodes, lecanora conizaeoides, l. pulicaris, lichenoconium erodens (on a thallus of lecanora conizaeoides), pseudevernia furfuracea, scoliciosporum chlorococcum and xanthoria polycarpa. two of them, lichenoconium erodens and xanthoria polycarpa, were recorded exclusively on this substrate. the recorded biota includes 11 species threatened with extinction in poland (19.6%), including two endangered species (en): lecanora albella and usnea subfloridana, four vulnerable species (vu): biatora efflorescens, ochrolechia androgyna, ramalina farinacea and usnea filipendula, and five near threatened species (nt): bacidina arnoldiana, evernia prunatri, graphis scripta, pertusaria coccodes and vulpicidia pinastri. four of them are threatened in north-eastern poland, including one endangered species: lecanora albella, two vulnerable species: bacidina arnoldiana and biatora efflorescens, and one data deficient lichen (dd): fuscidea arboricola (fig. 4). ten (17.9%) of the species given are protected in poland, including one partially protected (evernia prunastri) and 9 fully protected (melanelia fuliginosa, parmelia saxatilis, parmeliopsis ambiqua, platismatia glauca, pseudevernia furfuracea, ramalina farinacea, usnea filipendula, usnea subfloridana, vulpicidia pinastri). discussion quercus rubra is mostly a large tree with a thick, straight, regular trunk and a wide, spreading crown. outer bark in young red oaks, up to 40 years old, is smooth, steel-grey or dark grey, and furrowed or with flat-topped ridges in old trees. quercus rubra differs from native oaks (q. robur, q. petraea) both by the habit as well as the structure and thickness of outer bark. the trunk of quercus robur, which tends to branch out low above the ground (massive lateral boughs), is covered with thick, deeply furrowed, dark brown outer bark. both these features as well as other factors influence species composition of lichens associated with this phorophyte. these differences are emphasised by b a r k m a n (1969), who classifies quercus rubra along such porophytes as fagus sylvatica and young specimens of quercus robur and q. petraea. chemical properties of fagus sylvatica periderm (low ph) are similar to those of quercus robur and q. petraea; it differs from them, however, by the bark relief and the crown shape. these differences are not as pronounced in the case of red oak and beech. the lichen biota of both oak and beech has not been studied extensively in poland. r u t k o w s k i and k u k w a (2000) list 88 taxa, including 51 beech epiphytes and 77 oak epiphytes, in their analysis of the lichen biota of beech trees (fagus sylvatica) and native oaks (quercus robus and q. petraea) in 24 sites in northern poland. the list comprises 40 taxa common for both trees (45.4%). lichens of red oak 325 as these findings show, the lichen biota of red oak seems to be quite diversified and is comparable with native oaks, and especially beech. it should be stressed, however, that the study by r u t k o w s k i and k u k w a (2000) does not fully reflect species differentiation of its epiphytes in northern poland, particularly oak. it only includes a small number of sites in north-eastern poland, where, according to c i e ś l i ń s k i (2003a), 234 lichens of this phorophyte were recorded. c i e ś l i ń s k i (2003a), however, does not differentiate oak species, and the findings may concern various taxa. therefore, the information provided by r u t k o w s k i & k u k w a (2000) is of greater value in the assessment of lichen biotas of native oak species and beech and red oak. the similarity of lichen biotas (jaccard’s coefficient) of native oaks and beech, given in the study by r u t k o w s k i and k u k w a (2000), and the lichen biota of red oak is 32% for red oak and native oaks, and is slightly higher and equals 34% for red oak and beech. the total, aggregated number of epiphytes of oak and beech equals 109, including 22 taxa recorded exclusively on the bark of quercus rubra. the provided data show that the phorophytic profile of quercus rubra significantly differs from that of other, native oak species. the species has its own specific lichen biota which consists of many interesting taxa. as a result of the review of the polish lichenological literature, additional 19 lichen species reported from the bark of red oak were recorded. of those, 11 taxa were not observed in the olsztyn lake district. the list of lichens of red oak in poland comprises in total 63 lichen species and 4 species of lichenicolous fungi. this biota should be considered to be rich, especially as its differentiation appears to be similar to that recorded in the areas of the native range of this phorophyte. the number of taxa colonising the bark of quercus rubra in its native occurrence range is small although the fact that its trunks are often abundantly covered by lichens is stressed (cf. s t u b b s 1989; m a y 2001). h a l e (1955) reports 45 lichen species from the southern part of wisconsin, and c u l b e r s o n (1955) reports 41 species from the northern part of the state. s t u b b s (1989) presents 32 taxa (15 fruticose and foliose and 17 crustose) from the sites in main. the number of lichens of this important phorophyte in this region usually does not exceed 10 in numerous local studies on the north-western part of north america (h y e r c z y k 1996, 1998; m a y 2001). as the surface area covered by deciduous forests of the class querco-fagetea, associated with the richest lichen biotas in lowland regions, has been declining (c z y ż e w s k a 2003), quercus rubra, a phorophyte characterised by a broad ecological scale and potentially rich lichen biota, may play an important role in the preservation of lichen species diversity in forest phytocoenoses. the biocoenotic function of red oak, underestimated so far, may be of certain use in cultivated forests located outside or on the limit of the native occurrence of beech. special attention should be paid to the occurrence of lecanora albella, a species belonging to a group of lowland old-growth forest indicators, on the bark of red oak (cf. c i e ś l i ń s k i , c z y ż e w s k a , fa b i s z e w s k i 2003). the phorophyte has been reported on the bark of red oak both in the olsztyn lake district and gdańsk pomerania (ty s z k i e w i c z 1935). according to fa ł t y n o w i c z (1991), lecanora albella is in particular associated with beech bark in western pomerania. it occurs on a variety of deciduous species in north-eastern poland (c i e ś l i ń s k i 2003a) and exemplifies ecological vicarism (cf. b a r k m a n 1969; fa ł t y n o w i c z 1991; z a l e w s k a 2000). in the olsztyn lake 326 d. kubiak district, the species was recorded in the city forest in olsztyn where many other lichen lowland old-growth forest indicators also occur (k u b i a k 2005). conclusions 1. the epiphytic biota of quercus rubra as a phorophyte is characteristic, diversified and rich, and it comprises 63 lichen species and 4 species of lichenicolous fungi. 2. species of rare lichens, both threatened with extinction (lecanora albella, melanelia olivacea, ochrolechia androgyna, usnea subfloridana) and differentiated very rarely (fuscidea arboricola, f. pusilla, lecidella subviridis, ropalospora viridis), were recorded on the bark of red oak in poland. 3. as well as having many biocoenotic functions, red oak may play an important role in preserving lichen biodiversity in the forest environment in poland. acknowledgements: the author would like to thank dr. martin kukwa, department of plant taxonomy and nature protection, university of gdańsk, for his kind help in determining selected lichen specimens. he would also like to thank the anonymous reviewer for valuable comments and remarks. references a l v a r e z j., c a r b a l l a l r. 2000. flora liquenica sobre quercus robur l. en galicia (n w espana). cryptogamie, mycol. 21 (2): 103 117. b a r k m a n j. 1969. phytosociology and ecology of cryptogamic epiphytes. van gorcum, assen. b o r a t y ń s k a k . 1979. dąb bezszypułkowy (quercus petraea (mat.) liebl.) w północno wschodniej polsce. arboretum kórnickie 24: 69 85. b o r a t y ń s k i a. 1979. występowanie jaworu (acer pseudoplatanus l.) w polsce. arboretum kórnickie 24:19 67. b r o w i c z k . 1953. dęby uprawiane w polsce. rocznik sek. dendrol. 9: 86 88. b u g a ł a w. 1991. drzewa i krzewy dla terenów zieleni. pwril, warszawa. c i e ś l i ń s k i s. 2003 a. atlas rozmieszczenia porostów (lichenes) w polsce północno wschodniej. phy tocoenosis 15 (n.s.). suppl. cartogr. geobot. 15. c i e ś l i ń s k i s. 2003 b. czerwona lista porostów zagrożonych w polsce północno wschodniej. monogr. bot. 91: 91 106. c i e ś l i ń s k i s., c z y ż e w s k a k., f a b i s z e w s k i j. 2003. czerwona lista porostów wymarłych i za grożonych w polsce. monogr. bot. 91: 13 49. c i e ś l i ń s k i s., to b o l e w s k i z . 1988. porosty (lichenes) puszczy białowieskiej i jej zachodniego przedpola. phytocoenosis 1 (n. s.), suppl. cartogr. geobot. 1: 1 216. c o u l b e r s o n w. l. 1955. the corticolous communities of lichens and bryophytes in the upland forest of northern wisconsin. ecoll. monogr. 25 (1): 215 231. c z a r n o t a p., k u k w a m . 2003. some sorediate lichens and lichenicolous fungi new to poland. gra phis scripta 15: 24 32. c z y ż e w s k a k. 2003. ocena zagrożenia bioty porostów polski. monogr. bot. 91: 241 249. d a n i e l e w i c z w., p a w l a c z y k p. 2006. rola dębów w strukturze i funkcjonowaniu fitocenoz. 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(in:) m. ł a w r y n o w i c z , u . wa r c h o l i ń s k a (eds). rośliny pochodzenia amerykańskiego zadomowione w polsce. łódzkie tow. nauk., łódź: 97 150. h y e r c z y k r. d. 1996. the lichen flora of putnam county, illinois. transactions of the illinois state academy of science 89 (3): 143 156. h y e r c z y k r. d. 1998. the lichen flora of the st. charles park district natural areas. transactions of the illinois state academy of science 91 (4): 123 133. j u t r z e n k a tr z e b i a t o w s k i a., f e n y k m. a. 2001. wpływ klimatycznych czynników borealnych na kształtowanie się zbiorowisk leśnych polski północno wschodniej. acta bot. warmiae et masuriae 1: 25 49. k o n d r a c k i j . 1998. geografia regionalna polski. pwn, warszawa. k r ó l s. 1967. dąb czerwony quercus rubra l. w warunkach środowiska leśnego zachodniej polski. ptpn, prace kom. nauk roln. i kom. nauk leśn. 21: 419 482. k u b i a k d. 2005. lichens and lichenicolous fungi of olsztyn town (ne poland). acta mycol. 40 (2): 293 332. k u k w a m . 2006. nowe stanowiska rzadkich i interesujących porostów na pomorzu gdańskim. iii. acta bot. cassub. 6 (in press). k u k w a m., d i e d e r i c h p. 2005. monodictys epilepraria, a new species of lichenicolous hyphomycetes on lepraria. lichenologist 37: 217 220. m a t u s z k i e w i c z j. m. 2001. zespoły leśne polski. pwn, warszawa. m a y p. f. 2001. lichen survey of mount everett summit, southwest berkshire county, massachusetts. htttp://www.mounteverett.org/studies/lichen.pdf m o d r z y ń s k i j., r o b a k o w s k i p., z i e n t a r s k i j. 2006. zarys ekologii. (in:) w. b u g a ł a (ed.). dęby. nasze drzewa leśne 11. instytut dendrologii pan, poznań kórnik: 411 474. o r a n g e a., j a m e s p. w., w h i t e f. j. 2001. microchemical methods for the identification of lichens. british lichen society, london. p o d b i e l k o w s k i z. 1995. fitogeografia części świata. ameryka, australia, oceania, antarktyda. pwn, warszawa r u t k o w s k i p. 1995. flora porostów na dębach w polsce w świetle dotychczasowych doniesień literatu rowych. materiały konferencji i sympozjów 50 zjazdu ptb, kraków: 336. r u t k o w s k i p., k u k w a m . 2000. materiały do znajomości flory epifitycznych porostów dębów i bu ków w północnej polsce. bad. fizjogr. pol. zach. 49: 207 215. s t u b b s c. s. 1989. patterns of distribution and abundance of corticolous lichens and their invertebrates associates in quercus rubra in main. bryologist 92: 453 460. s z y m a n o w s k i t. 1959. zagadnienia aklimatyzacji obcych drzew w polsce. ochr. przyr. 26: 261 319. to k a r z h. 1971. zbiorowiska leśne z udziałem buka (fagus sylvatica) w obszarze północno wschodniej granicy jego zasięgu. 1. acta biol. med. soc. sc. gedan. 15 (3): 227 274. tu m i ł o w i c z j. 1965. abies balsamea mill. i abies concolor lindl. et gord. w lasach pomorza wschod niego. rocz. sek. dendr. ptb 19: 151 159. tu m i ł o w i c z j. 1967. ocena warunków wprowadzenia niektórych obcych gatunków drzew w lasach krainy mazursko podlaskiej. 1. rocz. sek. dendr. ptb 21: 135 169. ty s z k i e w i c z j. 1935. badania nad występowaniem porostów nadrzewnych w lasach północno wschod niej części wyżyny kielecko sandomierskiej. planta polonica iii: 1 119. z a l e w s k a a. 2000. ekologia porostów puszczy boreckiej i jej obrzeży. studium bioróżnorodności. praca doktorska (msc.). uniwersytet warmińsko mazurski w olsztynie, olsztyn. z a l e w s k a a., fa ł t y n o w i c z w., k r z y s z t o fi a k a., k r z y s z t o fi a k l., p i c i ń s k a fa ł t y n o w i c z j. 2004. porosty puszczy rominckiej. stowarzyszenie „człowiek i przyroda”, suwałki. z a r ę b a r. 1978. puszcze, lasy i bory polski. pwril, warszawa. 328 d. kubiak z e d d a l. 2002. the epiphytic lichens on quercus in sardinia (italy) and their value as ecological indica tors. englera 24: 1 468. porosty dębu czerwonego quercus rubra w środowisku leśnym pojezierza olsztyńskiego s t r e s z c z e n i e w wyniku przeprowadzonych badań własnych oraz przeglądu krajowej literatury licheno logicznej odnotowano 63 gatunki porostów oraz 4 gatunki grzybów naporostowych występują cych na korze quercus rubra. podczas szczegółowych badań nad porostami epifitycznymi tego forofitu, przeprowadzonych na 16 stanowiskach w lasach pojezierza olsztyńskiego, stwierdzo no 56 taksonów, w tym szereg gatunków rzadkich. wyróżniona na obszarze pojezierza biota liczy 11 gatunków umieszczonych na krajowej „czerwonej liście” oraz 14 porostów objętych ochroną gatunkową. do szczególnie interesujących taksonów zaliczyć należy: fuscidea arbori cola, lecanora albella, lecidella subviridis, ochrolechia androgyna i o. turneri. 2014-01-01t11:44:52+0100 polish botanical society © the author(s) 2014 published by polish botanical society flammulina ononidis – first record in poland janusz łuszczyński,, bożena łuszczyńska, agnieszka tomaszewska, kinga sobaś, małgorzata kostrzewa and katarzyna grudzień department of botany, institute of biology, jan kochanowski university świętokrzyska 15, pl-25-406 kielce, jluszcz@ujk.kielce.pl łuszczyński j., łuszczyńska b., tomaszewska a., sobaś k., kostrzewa m., grudzień k.: flammulina ononidis – first record in poland. acta mycol. 49 (1): 79–85, 2014. the authors present a first record of flammulina ononidis in poland. this species was characterized in respect of macroand micromorphological features, which were illustrated with the original figures and photographs. the habitat conditions of the recorded site were also described. a short discussion concerning the similar species and the ecology and threats of presented fungus species is provided. key words: fungi, basidiomycota, nida basin, ononis spinosa, xerothermic grasslands introduction the genus flammulina numbers about 15 species, of which in europe according to horak (2005) five, petersen et al. (on-line) seven species, but when pérez-butrón and fernández-vicente (2007) described f. cephalariae the number of species is currently eight. in poland, up to now, were known only two species – f. velutipes and f. fennae (wojewoda 2003). in 2012 in poland the first site of another species of this genus namely f. ononidis was recorded. this taxon, mostly known from europe, was the first time described by arnolds (1977), in germany. the aim of this article is the presentation and description of f. ononidis first site in poland, with a detailed description of the macroand micromorphological features, ecology, distribution and threat. material and methods in november and december 2012 more than 50 flammulina ononidis fruitbodies were collected, and the preparations were made from both the fresh and dry material. the acta mycologica vol. 49 (1): 79–85 2014 doi: 10.5586/am.2014.007 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 80 j. łuszczyński et al. © the author(s) 2014 published by polish botanical society fruitbodies were dried for 24 hours in the oven in temperature 39°c. the microscopic features were observed with the light microscope using the standard chemical reagents, 10% koh and floxine. the spores were measured in the sample of 50 spores obtained from 5 fruitbodies. the drawings of the spores and hyphae, pileocystidia and ixohyphidia were made based on the original photographs taken during the microscopic observations. the dried fruitbodies were deposited in the fungarium of the faculty of mathematics and science, jan kochanowski university in kielce. results description of the fruitbody. the fruitbody cap in the mature phase is flat-convex or flat, 15–45 mm in diameter, shiny, in a fresh state viscid, hygrophanous, at the cap edge the gills can be seen through, and in the old fruitbodies the cap is distinctly striped. the edges are from yellow-brown to golden-brown, changing in the direction of top, firstly into bright orange, and later on the top into bright brown, cinnamon-brown, dark carmel to rust-brown (figs 1–4). the caps of young fruitbodies are semispherical, slightly convex, golden-brown, bright brown-orange, elastic. the stem measures 30–85 x 2.5–3.5 mm, cylindrical, ending at the bottom part root like or spindle-shaped. the surface velvety. in the upper part, towards the gills, almost white or from creamy to bright yellow. the stipe colour is changing downwards, from bright lemon-yellow, through bright brown, and chocolate brown at the base, particularly in the mature fruitbodies. the stem inside empty. the gills are bright, white-creamy, but near to the edge the cap with an orange tint, 3–5 mm wide, of different length, adnexed, moderately thicken, with a straight and complete edge. the flesh elastic, in the cap pale yellow, in the upper part of stem whitish, in the bottom part particularly at the base wax-yellow, the taste mild, the smell indistinct. the basidia narrowly club like, four-spores, 28–34 x 6.5–7 μm. the spore mass white, cream-white. the spores from wide eliptical to almost almond like with two large or numerous different size drops of the fat, colourless and smooth, (9.45)10.8– 13.5 x 4.7–6 μm in size (fig. 5). pleurocystidia thin-walled 40–65 x 11.5–17.5 μm, of different shape, club like elongated, eliptical, inversely club like. cheilocystidia numerous, similar to the pleurocystidia. pileipellis made of the ixohyphidia and pileocystidia. the ixohyphidia thick-walled, branched either tree like or coral like or not branched, measuring 30–75 x 3.5–6.7 μm (fig. 6). the pileocystidia narrowly lageniform, 68–135 x 8.1–12 μm, thick-walled, at the base sometimes bent like letter “s”. the stem surface covered with the caulocystidia and cylindrical thin-walled hyphae. the caulocystidia are similar to the pileocystidia but they are slightly wider and more swelled. the thin-walled hyphae are moniliform and slightly constricted towards the top, often with short terminal cells. the hyphae in the stem upper part thin-walled and colourless, in the bottom part thick-walled with brown pigment. the site description. the fruitbodies of flammulina ononidis were found in the nida basin, 1 km west of the pińczów town (square atpol fe 13), in the phytocenosis of the alliance cirsio-brachypodion, in inuletum ensifoliae and thalictrosalvietum, which cover the southern slope of the limestone hills referred to as the “góry pińczowskie”. the fruitbodies were growing singly and in small groups on the flammulina ononidis – first record in poland 81 © the author(s) 2014 published by polish botanical society figs 1–4. flammulina ononidis on xerothermic grassland (photos 1–3 by k. sobaś, photo 4 by a. tomaszewska). 1 3 2 4 82 j. łuszczyński et al. © the author(s) 2014 published by polish botanical society soil, in the immediate neighbourhood of ononis spinosa. the role of ononis spinosa in the mentioned grasslands is particularly significant considering its quantitative participation. these are the phytocenosis where ononis spinosa covers the patches in the density from 20% to 30%. the areas of these communities up to not long ago were still grazed by the household cattle. presently the grazing since five years was completely ceased. the composition and the plant species participation in the investigated communities are illustrated by the phytosociological relevés. inuletum ensifoliae, ch.ass.: carlina onopordifolia +.2, inula ensifolia 5.5, linum hirsutum +; others: achillea pannonica +, adonis vernalis +, asperula cynanchica +, avenastrum pratense +, brachypodium pinnatum 1.1, carlina vulgaris +, centaurea scabiosa +, carex humilis 2.2, euphorbia cyparissias +, festuca rupicola +.2, galium verum 1.2, melampyrum arvense 1.1, ononis spinosa 2.2, peucedanum cervaria 2.2, ranunuculus bulbosus +, salvia pratensis +, sanguisorba minor +, seseli annuum +, thesium linophyllon +, thymus pannonicus +.2, th. marschallianus 1.2. thalictro-salvietum: elymus hispidus subsp. hispidus 2.2, adonis vernalis 1.1, agrimonia eupatoria 1.2, allium oleraceum +, anemone sylvestris 1.2, brachypodium pinnatum 2.3, carex humilis +.2, coronilla varia +, erigeron acer +, euphorbia cyparissias +.2, festuca rupicola +.2, fragaria viridis +.2, galium verum 2.3, geranium sanguineum +, gypsophila fastigiata 1.2, medicago falcata +, ononis spinosa 3.3, plantago lanceolata +, salvia pratensis 3.3, salvia verticillata 1.2, seseli annuum +, thalictrum minus 2.2, thesium linophyllon 1.2. the fruitbodies were observed and collected from november 2012 till january 2013. it is interesting that the mycelium was still producing new fruitbodies in january, despite of earlier frosts up to -150c, at the turn of november and december 2012. general distribution. flammulina ononidis is known first of all from the european countries. it was recorded up to now from: austria (krisai-greilhuber 1999), croatia (tkalčec et al. 2005), czech republic (antonín 2006), belgium (walleyn, vandeven 2006), denmark (petersen, vesterholt 2003), france (anonymus 1, 2, online), germany (arnolds 1977; krieglsteiner 1978), hungary (babos 1968; cetto 1994), estonia (urbonas et al. 1986), italy (petersen et al., on-line), russia (petersen et al., on-line), slovakia (ripková et al. 2008) and turkey (pekşen, karaca 2003). fig. 5. flammulina ononidis: spores. scale bar = 10 μm. fig. 6. flammulina ononidis: a – pileocystidia, b – ixohyphidia. scale bar = 10 μm. flammulina ononidis – first record in poland 83 © the author(s) 2014 published by polish botanical society discussion flammulina ononidis is the species mainly connected with ononis spinosa. the majority of until now published data refer to this plant species. however, it is interesting that, urbonas et al. (1986) collected the fruitbodies of f. ononidis on trifolium pratense. perhaps it is possible to collect f. ononidis fruitbodies on the other fabaceae family plants. it is curious that redhead and petersen (1999) in california, found f. velutipes var. lupinicola on lupinus arboreus. whereas pérez-butrón and fernández-vicente (2007) described from the northern spain a new species f. cephalariae of which the mycelium were developing on the roots of cephalaria leucantha (dipsacaceae). these examples provide an interesting perspective on the ecology of some species of the genus flammulina and their connection with the host plants. there is a striking morphological similarity of some flammulina species and this can pose a quite serious problem when identifying species in the field. distinctive differences between the species are marked at the level of the microscopic structures, in particular of the features referring to the structure of the cap skin and the size of spores. in table 1 are presented the spores sizes for the selected taxa of flammulina genus. the fruitbodies of these taxa morphologically are very similar. the spores size of flammulina ononidis collected in pińczów are almost precisely the same as those presented by arnolds (1977) and bas (1983). information referring to the ecology of f. ononidis are also concurrent with our observations referring to the investigated site in pińczów. according to antonín (2006), bas (1995), hagara et al. (2005), klán (1978), (kotlaba 1995), ripková et al. (2008) it is a species which prefers the habitats on the limestone soils in the plant communities belonging to the alliances bromion erecti and cirsio-brachypodion pinnati from festuco-brometea class. a very important element referring to the habitat demands is ononis spinosa as the host plant. this plant prefers dry, sufficiently table 1 comparison of spore size of the similar species from the flammulina genus species authors size of spores q av. q plant host f. ononidis arnolds (1977) (7.5) 8.5–13 (14) x (4) 4.5–6 . . ononis spinosa f. ononidis bas (1983) (7.5) 8.5–12.5 (14) x (4) 4.5–5.5 (6) (1.6) 1.7–2.45 1.9–2.3 ononis spinosa f. ononidis horak (2005) 8.5-13 x 4.5-5 . . ononis spinosa f. ononidis ripková (2008) (8.6) 9.4–10.6 (11.2) x (4.1) 4.3–5.2 (6) (1.76) 1.94–2.28 (2.43) 2.11 ononis spinosa f. velutipes var. lupinicola redhead and petersen (1999) 7–14.8 x 3.7–6.6 av. 4.5–5.4 . . lupinus arboreus f. cephalariae pérez-butrón and fernándezvicente (2007) (9.2) 12–16.8 (17) x (5) 5.6–7.6 (8) 2 cephalaria leucantha f. velutipes petersen et al. (on-line) 6–9.5 x 3–4 . 2–2.3 hardwood trees, mainly salix, populus, alnus f. elastica petersen et al. (on-line) 8–11.5 x 3–4 . 2.5–3 hardwood trees, mainly salix 84 j. łuszczyński et al. © the author(s) 2014 published by polish botanical society sunny habitats, growing on different soils from acid and rather loose sands to the soils more heavy and rich in caco3. in poland it is rather rare species, mostly connected with the xerothermic habitats. according to the cited authors f. ononidis was also found on the limestone sands of dunes and mentioned from the calcareous dunes and river dikes. endangerments. flammulina ononidis belongs to the endangered fungi in austria (krisai-greilhuber 1999), croatia (tkalčec et al. 2005), czech republic (holec, beran 2006) and germany (benkert et al. 1996). in poland the endangered problem for f. ononidis is similar to that in the above mentioned countries. this results from the discontinuation of traditional forms of exploitation of the xerothermic grasslands. as a result of this the grasslands are subjected to succession and are overgrown by the shady shrubby brushwoods. an effective form of both the fungus and hose plant protection is an active protection of the habitats and the types of communities, which are conducive for development of f. ononidis and ononis spinosa. the effective protective interventions should be rather intensive grazing with the home animals and cutting out the appearing brushwoods. references anonymus 1 (on-line). la modernisation des zones naturelles d’intérêt ecologique, faunistique et floristique en région nord pas-de-calais. http://www.nord-pas-de-calais.ecologie.gouv.fr/img/pdf/informations_generales-2.pdf anonymus 2 (on-line). flammulina ononidis arnolds. http://www.pharmanatur.com/mycologie/flammulina%20ononidis.htm antonín v. 2006. flammulina ononidis arnolds. 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(in:) h. niklfeld (ed.). rote listen gefährdeter pflanzen österreichs, bundesministerium für umwelt, jugend und familie, p. 229-266, graz. pérez-butrón j.l., fernández-vicente j. 2007. una nueva especie de flammulina p. karsten, f. cephalariae (agaricales) encontrada en españa. revista catalana de micologia 29: 81-91. flammulina ononidis – first record in poland 85 © the author(s) 2014 published by polish botanical society pekşen a., karaca g. 2003. macrofungi of samsun province. turkish j. bot. 27: 173-184. petersen r.h., hughes k.w., redhead s.a. (on-line). the genus flammulina, a tennessee tutorial. http:// www.bio.utk.edu/mycology/flammulina/default.html petersen j.h, vesterholt j. 2003. de danske svampenavne – en kommenteret navneliste. 76 p. ebeltoft. http://www.mycokey.com/mycokeydk/dkkeyspdfs/dedanskesvampenavne.pdf redhead s.a., petersen r.h. 1999. new species, varieties and combinations in the genus flammulina. mycotaxon 71: 285-294. ripková s., adamčík s., kučera v. 2008. flammulina ononidis – a new species for slovakia. czech mycol. 60 (2): 221-230. tkalčec z., mešić a., matočec n. 2005. crveni popis gljiva hr. http://www.dzzp.hr/dokumenti_upload/20100414/dzzp201004141250262.pdf urbonas v., kalamees k., lukin v. 1986. conspectus florum agaricalium fungorum (agaricales s. l.) lithuaniae, latviae et estoniae (materies 1778–1984 annorum). vilnius, 138 p. walleyn r., vandeven e. (eds). 2006. standaardlijst van basidiomycota en myxomycota van vlaanderen en het brussels gewest. 143 p. brussel. www.inbo.be/docupload/3062.pdf flammulina ononidis – pierwsze stanowisko w polsce artykuł prezentuje nowy dla polski gatunek grzyba flammulina ononidis. gatunek ten został scharakteryzowany pod względem jego cech makroi mikroskopowych, a także warunków siedliskowych, ze szczególnym uwzględnieniem składu fitocenoz w których wyrastał. nowe stanowisko grzyba zostało znalezione na wzgórzach kserotermicznych koło pińczowa w płatach inuletum ensifoliae i thalictro-salvietum (klasa festuco-brometea), gdzie wyrastał na korzeniach ononis spinosa. przedstawiono krótką dyskusję dotyczącą gatunków podobnych oraz ekologii i zagrożeń prezentowanego gatunku grzyba. 2014-06-30t22:29:49+0200 polish botanical society behaviour in cultures and habitat requirements of species within the genera loreleia and rickenella (agaricales) andreas bresinsky and angelika schötz institute of botany, regensburg university d 93040 regensburg, abresinsky@t online.de b r e s i n s k y a . , s c h ö t z a . : behaviour in cultures and habitat requirements of species within the genera loreleia and rickenella (agaricales). acta mycol. 41 (2): 189 208, 2006. the term eco geogram is introduced for surveying (in logical order) ecological and geographical data connected with fungal species. the database pilzoek was established for that purpose. eco geograms are provided in this paper as an example for data retrieval from pilzoek concerning the agarics loreleia marchantiae, l. postii, rickenella swartzii and r. fibula. the potential degree of endangerment is discussed in regard to habitat requirements. european species of loreleia are not regarded to be endangered in central europe, although the risk to get threatened, because of low frequency of fruit body occurrence and quite a narrow substrate specialization, could be higher than in case of rickenella fibula and r. swartzii. cultures of rickenella fibula, r. swartzii, loreleia marchantiae (= gerronema daamsii) and l. postii were investigated in regard to pigment accumulation, chlamydospore formation and some other characters. key words: eco geogram, rickenella, loreleia, habitats, substrates, cultures, endangerment introduction the genera loreleia and rickenella emerged taxonomically from the genus gerronema in its broader sense, as it was originally interpreted by s i n g e r 1986. various contributions to the generic concept brought evidence to the assumption that gerronema in the sense of s i n g e r (1986) represents an artificial assemblage of species. gerronema s. l. includes species with quite different requirements in regard to habitats and substrata. some species are bryicolous (being transferred to loreleia and rickenella), some are lignicolous (being transferred to chrysobostrychodes), some are lichenized (being transferred to lichenomphalia), some are humicolous and finally some imply unknown or badly known habitat relations. for understanding the evolution and for being aware of the existence and of the endangerment of fungi, as well, it seems to be necessary to investigate substrate and habitat requirements in correlation to the phylogenetic position of the species in focus. in this contribution habitat requirements of selected fungal species will be arranged following a scheme acta mycologica vol. 41 (2): 189-208 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 190 a. bresinsky and a. schötz which has been designed for an ecological and geographical database for fungi (pilzoek). a quite interesting question is, to what degree the endangerment of species is correlated with specialized habitat requirements. the chance for survival of a species depends largely on the availability of suitable habitats and substrates for growth and fructification. if such environmental prerequisites are restricted in any sense and if they are at the same time quite specialized, the risk of extinction might be much higher than in case of ambiguous ecological demands. that means that the endangerment of fungal species is largely to be understood in respect to their habitat relationships. in case of the macromycetes this fact is most important, since their predominating mycelial condition in the field is more or less invisible to the observer’s eyes. the risk of endangerment is much higher in stenoecious species, which are restricted on very special and maybe vulnerable habitats, than it is in euryoecious species, which are able to survive under a variety of different environmental conditions, at least if some of them turn out to be well distributed, stable and not decreasing (b r e s i n s k y et al. 2005). the possible correlations between the existing or not existing degree of endangerment on one side, and habitat specialization on the other side, will be discussed in this paper along the example of four bryicolous gerronema s.l. – species assigned nowadays to the genera loreleia and rickenella, respectively. methods the culture experiments with species of loreleia and rickenella have been performed by angelika a c h h a m m e r ( s c h ö t z ) (1986) in the lab of the senior author. the strains of fungi which have been cultivated list as follows: gerronema (chrysobostrychodes) chrysophyllum (fr.) singer, obtained from fresh material, germany, bavaria, bayerischer wald, mauther wald, 05.09.1984, leg. a. bresinsky. gerronema daamsii marxmüller & clémençon: culture-number regensburg455, kortenhoef, nl. submitted by j. daams, the netherlands. gerronema strombodes (berk. et mont.) singer , obtained from fresh material, germany, bavaria, weiherholz, ascholding, 18.08.1984, leg. a. bresinsky et a. einhellinger. loreleia (gerronema) marchantiae (singer et clémençon) redhead et al.: culture-number regensburg-442. obtained from the culture collection in hannover, germany, number 7826. loreleia (gerronema) postii (fr.) redhead et al., culture obtained from fresh material (spores): germany, bavaria, valley of weiße laber, e of matzenhof, swamp with polemonium caeruleum, 13.05.1984, leg. o. mergenthaler. rickenella fibula (bull.: fr.) raithelh., cultures obtained from fresh material (spores): germany, bavaria, berchtesgaden, hintersee, on aulacomnium palustre, 06.06.1984, leg. a. bresinsky & w. schmid-heckel. germany, bavaria, geisental, n regenstauf, on calliergonella cuspidata, 06.08.1984, leg. n. luschka. germany, bavaria, paintner forst, lkr. kelheim, rothenbügl, on dicranella heteromalla, 10.10.1984, leg. g. laaser & b. meixner. rickenella swartzii (fr.) kuyper, culture obtained from fresh material (spores): germany, bavaria, deusmauer, lengenfeld, alder swamp forest, on dicranella heteromalla, plagiothecium latebricola, 03.06.1984, leg. w. paulus. behavior in cultures and habitat 191 culture media. moser-b-medium modified (m o s e r 1960; p r i l l i n g e r , s i x 1983). medium for cellulose-test (ta n s e y 1971): moser-b-medium without maltose, however, with glucose (4g / 10g) and cellobiose (10 g / 4 g). addition of 5.0 g swelled cellulose in form of a diluted suspension. water was added as much as necessary to obtain a total volume of 1 liter; ph 6. no agar was added for preparation of a liquid medium. casein-mb-medium: a) moser-b-medium. water was added as much as necessary to obtain a volume of 700 ml; ph 6. b) 30 g powdered skim milk was diluted in 300 ml aqua dest. a) and b) were autoclaved separately and then, at a temperature of 45o c, united (a n d e r s o n 1962; we y l a n d 1970). gujacol-mb-medium: 0.05 g gujacol was added to moser-b-medium (b o i d i n 1951; l y r 1958). test for cellulase activity. the medium for the cellulase test looks originally dull because of the nature of swelled cellulose; cellulase activity of inoculated fungus gets visible by a clarified zone around the inoculum. cellulase activity was evaluated semi-quantitatively by measuring the diameter of the clarified zone. confrontation tests with marchantia. thalli of marchantia, or parts of it (clones), were washed and sterilized with ethanol (96%) and then placed on a medium (see above “culture media”) in petri dishes. the rhizoids were exposed to the surface of the medium which had been inoculated with the fungus in advance. later, inocula were also placed close to or on the thallus of the liverwort. then the petri dishes (containing the liverwort and the fungus) were kept in a humid chamber. in a modified arrangement marchantia was grown in flower pods and then directly inoculated with the fungus. the inoculated pods with marchantia were kept in a humid chamber which was, once a day for one hour, ventilated. data base pilzoek. the ecological data connected with the species treated here have been collected by means of the data base pilzoek (b r e s i n s k y , d ü r i n g 2001). the input of data is at its preliminary stage. so far it includes, as a matter of fact, only a restricted number of the available data from germany and its neighbouring countries. the database will be completed steadily by the senior author of this paper. most valuable data came from publications on mycocoenological interrelationships which had been elaborated through mycologists in poland, in the tradition of intensively and long lasting efforts in that country (going back to s k i r g i e ł ł o , n e s p i a k and other workers; for references see wo j e w o d a 2003 ł a w r y n o w i c z et al. 2004, a most valuable survey of mycocoenological literature from poland). at present about 11.000 sets of data are incorporated in the database pilzoek. each data set represents a linkage of fungal species with ecological / geographical factors (one fungus species with many factors or, respectively, one factor with many fungal species). the current over-all-number of connections established in the database is estimated to a rate of 160.000. the data, which are included and stored in the database, can be retrieved by different options. it is possible to start inquiries for ecological and geographical factors linked with a selected species in focus (i. e. pre-selection of one fungal species results in a list of ecological and geographical factors connected with that species). another possibility is to ask for lists of fungal species connected with a selected ecological or geographical factor (i. e. pre-selection of an ecological or a geographical factor results in a list of fungal species connected with that factor). the database is accessible free of charge for everyone via internet (http://www.pilzoek.de; for details see b r e s i n s k y , d ü r i n g and a h l m e r 2005). 192 a. bresinsky and a. schötz the final design of the database pilzoek and its presentation in the internet was made possible through a grant of the german ministry of education and research in the context of the biolog-program (biolog biodiversity and global change). results the available ecological data in regard to the species loreleia marchantiae, loreleia posti, rickenella swartzii and rickenella setipes are listed and discussed. geographical data will be mentioned on the european and world-wide scale only; regional distributional patterns are neglected in this paper. the survey of speciesbound ecological characters according to a scheme of order will be named ecogram (in analogy to the terms program, telegram, etc.) and for the survey of geographical (distributional) traits the term geogram is introduced here; for both together, the term eco-geogram is applied. the listed code numbers are identical with those which are implemented in the data base pilzoek. the hierarchy of code numbers reflect a given scheme of order for the presentation of data (survey in b r e s i n s k y , k r e i s e l and p r i m a s 1995) which makes additions of new records or a search for special information easier. references on mentioned factors (f. e. in the discussion) may be given in short form by code numbers only. the data will be presented in different sections (a-i) which are part of the data presentation program of the data base pilzoek; in the context of the species treated here, some of the sections will be found omitted (f. e. e, h). the sources of the data presented in the eco-geograms are indicated by the names of authors and contributors. the full citations of sources (of incorporated data) are given in the database pilzoek and are accessible through the above mentioned internet address; therefore, these will be omitted here. abbreviations used for countries / regions: a = austria, b = belgium, ch = switzerland, cro = croatia, d = germany, dk = den mark, est = estonia, far = faeröer, f = france, gb = great britain, h = hungary, i = italy, isl = island, lat = latvia, lit = lithuania, n = norway, nl = netherlands, p = portugal, pl = poland, ru = russia; s = sweden, sc = sicily, sd = sardinia, sf = finnland, slq = slovakia, sln = slovenia, spi = spitsbergen / svalbard, tch = tschecho slovakia, tq = turkey, ukr = ukrainia. indications within brackets like < > mean: not commonly realized in nature or even ques tionable. loreleia marchantiae (singer and clémençon) redhead et al. syn.: gerronema marchantiae singer et clémençon, omphalina marchantiae (singer, clé mençon) norvell, gerronema daamsii marxmüller et clémençon. observations. mycelial cultures of loreleia marchantiae and l. postii, as well, can be established rather easily from fresh material (a c h h a m m e r 1986; see methods). cultures of both species exhibit quite a high cellulase activity which can be interpreted as necessary in order to penetrate the cells of the bryophytes and to live as parasites. if mycelia of l. marchantiae are confronted with thalli of marchantia polymorpha, the hyphae of the fungus penetrate into the host and are found then predominantly within the tuberculate rhizoids (fig. 1b); later partly also within the tissue of the thallus (fig. 1c). the hyphae of l. marchantiae develop on ( on rh), or, respectively, within the rhizoids ( in rh) of marchantia different structures: appressoria (on rh), penetration hyphae (in rh), vesicular haustoria (in rh), chlamy behavior in cultures and habitat 193 fig. 1: a c) loreleia marchantiae (= daamsii) and d) loreleia postii (cf. marchantiae) in confrontation with marchantia: a) hyphae within rhizoids of marchantia; b) appressoria (a), haustoria (h), penetrating hyphae (p) formed in contact with tuberculate rhizoids of march antia; c) haustorium (h) in a cell of marchantia; d) mycelium penetrating a rhizoid (left side) and within a rhizoid (right side) of marchantia. 194 a. bresinsky and a. schötz dospores (in rh) and oidia (in rh). since no damage or restricted growth of the infected thallus can normally be observed under natural conditions, some kind of symbiotic interrelationship between both organisms might exist, in analogy to (endo-) mycorrhizal interactions in higher plants. in fresh material which had been collected in the botanical garden in cracow, poland, g u m i ń s k a and m i e r z e ń s k a (1992) observed an association of l. marchantiae not only with smooth and tuberculate rhizoids of marchantia but also with coenobia of cyanobacteria belonging to the genera anabaena and nostoc. these cyanobacteria were not found in uninfected thalli of marchantia. in thalli infected by the fungus they were observed in the lower parts of the thallus chambers, between the rhizoids and at the base of the stipe. the authors conclude that the fungus mediates between cyanobacteria and liverwort through hyphal connections making accessible nitrogen compounds produced by the former. parasitism has to be excluded in the view of these authors. another interpretation is possible in the sense that l. marchantiae behaves as a weak parasite or as a saprobiont and, indeed, a record made by the senior author exhibited a necrotic area of marchantia polymorpha around the attachment of the stipe of the fungus (notes on the record in s c h m i d h e c k e l 1985). eco-geogramm for loreleia marchantiae (using database pilzoek): sources: gumińska & mierzeńska (1992); klán (1992); kühner & lamoure (1986); legon & henrici 2005; lüderitz 2003; schmid-heckel (1985); wöldecke (1998). for references see http://www.pilzoek.de a. nutrition: 0.1 bryicolous parasite (?), 0.32 bryicolous endotrophic mycorrhiza (?), 0.4 lichenized (?), 0.5 bryicolous saprobiont (?) b. substrates: 5.1 liverworts. 5.72 (living, dying, dead): conocephalum, lunularia, marchantia polymorpha, m. alpestris, m. paleacea! [d, gb, pl, tch]. 5.41 associated with cyanobacteria: anabaena, nostoc [pl]. c. habitats: 11.1 humid woods with maple (aceri-fagenion). 14.14 ash-alderand alder-ash-wods along brooks. 18.3 paths in woods. 19.2 park woodlands / parks. – 19.5 botanical gardens; outdoor areas. 21.0 paludal areas around springs / vegetation around headwaters. 22.3 transitional bogs. 29.2 shores / alluvial plant communities [nl]. 30.7 gardens. 31.1 ruderal sites / weed vegetation. 32.3 field-paths / meadow-paths. d. soil characters:45.2.7 rather humid, not wet. g. phenology, persistence and sociability of fruit bodies: < 52.1 spring >, 52.2 summer, 52.3 autumn. 53.2 short living fruit bodies. i. distribution: 59.11 europe, north: n, dk, sf. west: gb, f, nl, b. central: ch, d, a, tch, slq, pl, dk. 59.13 north america (alaska). 59.16 greenland. k. distribution and endangerment in central europe: 62.1 planar, 62.2 colline, 62.3 montane (1300 m; d), 62.6 alpine (2300 m; f). 65.0 endangerment: 65.4 rarity, latent endangered? comments. fruit bodies of loreleia marchantiae are observed only seldom. nevertheless the species might be distributed quite well and shall be expected at any place where thallose liverworts are forming expanded mats. the specialization on few genera of liverworts for fructification and the presumed interrelationship with cyanobacteria (g u m i ń s k a , m i e r z e ń s k a 1992) could be the reason for a potential endangerment of the fungus. on the other side, the occurrence of marchantia is behavior in cultures and habitat 195 favoured by human activities (greenhouse culturing, fire places, nitrification, application of herbicides) and it might be quite likely that l. marchantiae is able to follow its host everywhere. the fungus is “growing in disturbed habitats, e. g. soil treated with herbicides (l i s i e w s k a , b a l c e r k i e w i c z 1991), on mud along ditches, between bricks along roads etc.” (b a s et al. 1995). it has to be considered, that the fungus might be much more frequent and distributed in its mycelial stage than it is indicated by its rare fructification, thus being mostly not apparent to the observer. generally, rhizoids and thalli of liverworts have been found to be quite regular infected by fungal hyphae (n i c o l a s 1967). in case of l. marchantiae a higher humidity is apparently afforded to enable fructification of the hosted fungus. thus, fructification in countries and areas with humid climatic conditions is more likely to be expected (oceanic climatic regime) than under more dry conditions of a continental climate. maybe, that this is the reason why this species has been recorded as being not so rare in western europe (b a s et al. 1995) and why it is so far missing or rare in the eastern european countries. loreleia postii (fr.) redhead et al. syn.: gerronema postii (fr.) singer, omphalina postii (fr.) singer, l. marchantiae (singer et clémençon) redhead et al. pr. p. observations. the species in its broader sense has been reported to be bound to thallose liverworts (f. e. marchantia), peat mosses (sphagnum) and mosses (ceratodon, funaria, polytrichum), as well. the distinction of loreleia postii s. str. from l. marchantiae, in the sense of c l é m e n ç o n 1982, on behalf of the association with thallose liverworts on one side (in case of l. marchantiae) and with mosses on the other side (in case of l. postii s. str.) seems to be quite hypothetical at the moment. the discriminating morphological characters between both species are minute and intergrading (d e r b s c h 1977); moreover, it is not clear if any kind of correlation does exist between the choice of hosts (liverworts versus mosses) and genetic or morphological characters. in recent studies (k l á n 1992; b a s et al. 1995) the separation of both species is not based on different hosts rather than exclusively on morphological characters: the pileus in l. marchantiae is claimed to be not striate, in l. postii it is striate. other differing features are mentioned for the margin (crenulated in l. marchantiae; entire in l. postii), for the gills (distant and forked in l. marchantiae; close and not forked in l. postii) and for the spores with slightly differing, however, overlapping sizes. all these differences could simply reflect different developmental stages of one and the same species. the pigmentation and morphological appearance of cultures (l. marchantiae and l. postii) were very similar to each other. confrontation of dicaryotic mycelia of both species did not result in any kind of barrage neither in a marked confrontation zone. instead of such expected behaviour, the mycelia of both species intermingled with each other. in the area of first interspecific hyphal contacts quite a number of anastomoses between intermingling hyphae could be observed, much more than near inocula of both species. however, it could actually not be decided if the anastomoses were inter-specific or intra-specific. from these observations it might be suggested that l. marchantiae and l. postii are very close to each other, if not even conspecific. evaluating the experimental observations, one should be aware of the fact, that the results are as good, or as bad, as the strains being used were correctly identified 196 a. bresinsky and a. schötz ore not. it is possible that the strains which were cultivated as “postii” and “marchantiae” represented only one species, eventually only l. postii. such an identity would explain the observation that the mycelial culture of g. daamsii ( l. marchantiae) behaved quite different from those labeled as “postii” and “marchantiae”. another possibility explaining the different behaviour of g. daamsii (as compared to l. marchantiae and l. postii) could be a contamination of its culture with another fungal species. in this case, however, it is surprising enough that in confrontation tests the hyphae of g. daamsii were found to penetrate into the rhizoids of marchantia as one would expect it for true g. daamsii and l. marchantiae. at the time being, l. postii should be interpreted as a taxon which is bound to mosses (sphagnum, funaria etc.) and l. marchantiae as a taxon restricted on liverworts (this is in accordance with the interpretations of c l é m e n ç o n 1982 and h o r a k 2005). experimental studies, including dna-analysis, should make it possible, to reveal whether this provisional concept holds true or not in the future. eco-geogramm for loreleia postii (using database pilzoek): sources: horak 2005; klán 1992; kreisel 1987; krieglsteiner 1991; legon & henrici 2005; schroeter 1889; singer 1964; wöldecke 1998. for references see http://www.pilzoek.de a. nutrition: 0.1 bryicolous parasite (?), 0.32 bryicolous endotrophic mycorrhiza (?), 0.5 bryicolous saprobiont (?). b. substrates: < 5.1 liverworts [gb]; cf. loreleia. marchantiae? >, 5.2 mosses!, 5.3 peat mosses! [gb]. 5.72 (living): < lunularia, marchantia? >, ceratodon purpureus, funaria hygrometrica(!), polytrichum, sphagnum (!) [gb]. c. habitats: 13.0 ombrogenous bogs and associated plant communities. 16.12 spruce forests: 16.42 rich on bryophytes. 18.0 clearances and open areas within woods. 19.2 park woodlands / parks. 30.91 flower pods [gb]. 31.7 pits (sand-. loam-, clay-). 31.8 outdoor fire places [gb]. 34.1 greenhouses [nl]. 37.5.2.1.0 funarietum hygrometricae. d. soil characters: 40.4 sand, 40.9 peat [gb]. 44.3 rich on nutrients (nitrogen). 45.2 humid, mesic; 45.3 wet. i. distribution: 59.11 europe, north: n, dk, s, sf, lit, est. -west: gb, f, nl. central: d, tch, pl, dk. east: lit. south: i. 59.13 north america. k. distribution and endangerment in central europe: 62.2 colline, 62.3 montane, 62.5 subalpine, 62.6 alpine. 65.0 endangerment: 65.4 rarity, latent endangered? comments. the data concerning distribution of the species belonging to loreleia in europe are probably not always correct since separation between l. postii s. str. and l. marchantiae (including g. daamsii) is still quite obscure, depending on different interpretations of taxa. l. postii s. str. seems to be rather rare, however, eventually not endangered, since it is reported to occur also in man made habitats like burnt places (31.8) and on flower pods (30.91). however, it is not absolutely clear if all of these indications apply only on l. postii, and not partially also on l. marchantiae. low frequency of distribution and specialization on few habitats may cause indeed a higher risk of endangerment. if l. postii should exclusively be restricted to mosses as substrate, then the grade of endangerment might appear to be higher. on the other hand, some of the mentioned host species among the mosses are widely distributed and common: funaria hygrometrica is growing everywhere on fire places (31.8) and ceratodon purpureus is by no means rare. behavior in cultures and habitat 197 rickenella swartzii (fr.) kuyper syn.: r. setipes (fr.) raithelh., gerronema setipes (fr.) singer observations. spores of this species and of rickenella fibula, respectively, germinated on suitable media, but not reliable and not always to a high percentage (as it has been already stated by l a m o u r e 1979). the cultures of rickenella swartzii and r. fibula showed identical or very similar characters. the readiness of spores to germinate depended on the season. if spores were collected in early summer (july) or in late fall (october), then the readiness to germinate and the speed of growth of arising hyphae was much higher than in the case that spores had been obtained in the time between these months. the regular pigmentation of cultures was whitish, ochraceous, or brownish, varying in intensity and colour from strain to strain. in some cultures pigments similar to those of the fruit-bodies accumulated if the cultures were grown in the spring and kept in day light. cultures of r. swartzii (03-06-84) on casein-mb-medium exhibited violet pigments, and on gujacol-mb-medium violet and orange pigments. in other words, both components of pigmentation were evident which are typical for the carpophores. a transfer of the cultivated mycelia from liquid culture medium to a solid culture medium (in this case prepared by adding an extract of hypnum cupressiforme to the standard ingredients) increased the intensity of pigmentation sometimes. eco-geogramm for rickenella swartzii (using database pilzoek): sources: achhammer 1986; antonin, noordeloos 2004; arnolds 1977, 1981; babos 1989; bas et al. 1995, besl et al. 1982; bresinsky 2000; bujakiewicz 1973, 1979, 1982, 1986; courtecuisse 1994; derbsch, schmitt 1987; einhellinger 1964, 1976, 1977, 1982; gulden, torkelsen 1996; gumińska 1976; hallgrimsson 1981; hansen, knudsen 1992; haeupler et al. 1981; horak 1963; jahn et al. 1967; kalamees, vaasma 1981; kost 1984; kreisel 1957, 1970, 1987; krieglsteiner 1991, 2000; krieglsteiner l. 1999, 2001; kühner, lamoure 1986; lambinon et al. 1977; legon, henrici 2005; luschka 1993; malençon, bertault 1975; miller, farr 1975; neuhoff 1949; nuß 1999; paulus 1981; ricek 1989; schmid-heckel 1985; sedlmeir 1985; skirgiełło 1998; šmarda 1973; sonneborn, stangl, sedlmeir and geh 1987; urbonas, kalamees and lukin 1986; watling, rotheroe 1989; winterhoff 1993, 1993b, 1994, 2001b, 2002; wojewoda 2003; wöldecke kn. 1990; wöldecke 1998; wöldecke, wöldecke 1988; wöldecke 2001. for references see http://www. pilzoek.de a. nutrition: 0.1 bryicolous parasite (?), 0.32 bryicolous endotrophic mycorrhiza (?), 0.5 bryicolous saprobiont (?). b. substrates: 5.0 interrelationship with bryophytes as substrates: 2.33 bryophyte covered wood on the ground (abies, fagus). 5.2 mosses. 5.71/5.72 (living): atrichum undulatum, aulacomnium palustre, brachythecium albicans, brachythecium rutabulum, brachythecium starkei [pl], calliergonella cuspidata, campylopus, ceratodon purpureus, climacium dendroides, dicranella heteromalla, dicranum bonjeanii, dicranum scoparium, drepanocladus vernicosus, eurhynchium angustirete [pl], eurhynchium praelongum, eurhynchium striatum, hylocomium splendens, hypnum cupressiforme, isopterygiopsis, mnium hornum, orthodicranum montanum, philonotis, plagiomnium affine, plagiomnium cuspidatum, plagiomnium elatum, plagiomnium rostratum [pl], plagiomnium undulatum, plagiothecium latebricola, pleurozium 198 a. bresinsky and a. schötz schreberi, polytrichum formosum, polytrichum juniperinum, polytrichum piliferum, rhizomnium punctatum, rhytidiadelphus squarrosus, scleropodium purum, thuidium tamariscinum. c habitats: woods / copses: 10.0 beech woods, mixed beech forests (fagion silvaticae pr. m. p.): 10.11.01 galio odorati-fagetum, 10.11.02 hordelymo-fagetum. 10.12 beech woods with share of white wood fir (abies; dentario glandulosaefagenion) [pl]. 10.3.01 luzulo-fagetum. 11.0 mixed frondose woods without (dominating) beech: 11.1 humid mixed woods with maple or with ash: 11.1.03 sorbo ariae-aceretum pseudoplatani [pl]. 11.1.04 fraxino-aceretum pseudoplatani. 11.3 oak-hornbeam woods (carpinion betuli): 11.3.01 galio sylvatici-carpinetum, 11.3.02 stellario holosteae-carpinetum betuli. 11.5 mixed oak woods on acid soils (genisto tinctoriae-quercenion robori-petraeae): 11.5.01 holco mollis-quercetum, 11.5.03 genisto tinctoriae-quercetum petraeae. 12.0 conifer dominated woods (natural): 12.2 spruce woods outside bogs (piceion abietis): 12.2.03 homogyno-piceetum [ch]. 12.51.06 pyrolo-pinetum sylvestris . 13.0 ombrogenous bogs and associated plant communities: 13.32 woods on bogs with scots pine, 13.34 woods on bogs with birch: 13.34.01 vaccinio uliginosi-betuletum pubescentis. 14.0 woods of river lowlands and swamps: 14.1 flood-plain woods: 14.12.01 alnetum incanae. 14.13 softwood riparian forests, willow beds (salicion albae). 14.14 mixed woods with ash and alder along brooks: 14.14.03 carici remotae-fraxinetum, 14.14.04 stellario nemorum-alnetum glutinosae, 14.14.06 pruno-fraxinetum. 14.15 hard-wood floot plain forests: 14.15.01 querco-ulmetum minoris. 14.2 paludal forests: 14.21 alder swamp forests (alnion glutinosae): 14.21.01 carici elongatae-alnetum glutinosae, 14.21.03 caltha palustris-alnus glutinosa-community [pl]. 14.22 grey willow swamps: 14.22.03 salicetum auritae, 14.22.04 salicetum cinereae. – 15.0 mantles of woods / bushes / hedges: 15.1.03 pruno-ligustretum. 15.33 green alder bushes (adenostylion alliariae): 15.33.01 alnetum viridis. 16.0 man made forests / nonnatural wood communities: 16.1 coniferous forests: 16.11 pine forests: 16.111 scots pine forests, 16.11.01 pure scots pine forests. 16.12 spruce forests: 16.12.01 typical oxalis-spruce forests [tch], 16.12.12 nude spruce forests without herbaceous layer [tch], 16.12.25 spruce forests on basic (lime stone) soils. 16.14 larch forest plantations. 16.2 deciduous forests. 18.0 clearances and open areas within woods: 18.3 paths through forests, 18.31.16 calluno-sarothamnetum. 19.4 garden copses. outside woods and copses: 21.0 paludal areas around springs / vegetation around headwaters [a]. – 22.0 soligenous bogs / transitional bogs: 22.2 acidophilous, non-calcareous reed marshes with dwarf sedges (juncion acutiflori): 22.2.05 juncetum acutiflori [nl]. 24.0 dunes / dry grassland on sand: 24.13 grey dunes. 24.2 inland dunes and associated grassland communities (thero-airion, corynephorion canescentis): 24.2.05 airo caryophylleae-festucetum ovinae [nl], 24.2.06 thymo-festucetum ovinae [nl], 24.2.13 diantho deltoidis-armerietum elongatae. 25.1 subalpine / alpine grasslands on limestone. 25.4 snow pockets: 25.42.01 salicetum herbaceae. 25.54 calcareous fens (caricion davallianae). 27.0 grassland / drifts: 27.1 dry and medium dry grasslands: 27.12.03 gentiano-koelerietum. 27.23 matt-grass areas in lower (colline) elevations: nardo-callunetea [nl], juncion squarrosi [nl]. 28.0 meadows / pastures: arrhenatherion elatioris: 28.11.01 arrhenatheretum elatioris. 28.12.02 geranio-trisetetum. 28.13 rich behavior in cultures and habitat 199 pastures / trampling resistant grassland: 28.13.03 festuco-cynosuretum, 28.13.04 lolio-cynosuretum, 28.13.14 prunella vulgaris-plantago major-community. 28.14 oligotrophic grassland. 28.15 park lawns. 28.16 garden lawns . 28.21 moist and wet meadows (calthion): 28.21.02 angelico-cirsietum oleracei, 28.21.06 scirpetum sylvatici [nl], 28.21.10 bromo-senecionetum aquaticae [nl]. – 28.22 litter meadows (molinion caeruleae): 28.22.01 molinietum caeruleae, 28.22.03 cirsio tuberosimolinietum arundinaceae [nl]. – 29.2 shores / alluvial plant communities. 30.7 gardens. 30.91 flower pods. 32.3 field-paths / meadow-paths: plantaginetea majoris [nl]. fungal communities: 38.42.41 geastro (quadrifidi)-agaricetum semotae [tch] d. soil characters: 42.1 calcareous soils. 42.22 sand soils, siliceous sands. 43.0 acidity of soil: 43.1 acid, ph 4.1-4.8 [tch]; 43.2 neutral, ph 5.7-6.5; 43.3 basic, ph 6.6-7.5. 45.2 humid, mesic; 45.3.9 wet, water soaked, with poor aeriation. 45.1 dry, xeric. f. temperature and light at the stand: 50.2.3 cool, 50.4.7 warm. 51.1 shadow, 51.7 half light, 51.8 light. g. phenology, persistence and sociability of fruit bodies: < 52.1 spring >, 52.2 summer, 52.3 autumn, < 52.4 winter >. 53.2 short living fruit bodies. 54.1 single, 54.2 in groups. i. distribution: 59.11 europe, north isl, spi, n. dk, s, sf, lit, est. west: gb, f, b, nl. central: ch, d, a, tch, slq, pl, dk. east: h, ukr, lit. south: sd, i, cro. – 59.12 asia (kamchatka). 59.13 north america.59.15 north africa: marocco. k. distribution and endangerment in central europe: 61.3 alpid, 61.5 temperate, central european, 61.6 boreal. 62.1 planar, 62.2 colline-submontane, 62.3 montane (600-1000 m; d, pl, tch), 62.4 high-montane (1040-1100 m; d, pl), 62.6 alpine (2500 m; f). 65.0 endangerment: predominantly not threatened. comments. in case of rickenella swartzii the frequency of distribution in terms of observed carpophores in the field is much lower than in r. fibula. nevertheless, the species is most likely not threatened, because it grows and fruits in numerous and various habitats which are to a greater extent (so far) not endangered. the occurrence in man made habitats which sometimes are rich in nutrients demonstrates that the species has quite a high capability to survive the decrease of vulnerable, nutrient poor habitats. the readiness of spores to germinate and to establish mycelia under not absolutely fixed biotic and abiotic conditions is also in favour for quite a high probability to survive and to fruit in sufficient frequency. these conditions altogether contributed apparently to the evaluation of a not threatened species; it is f. e. not included in red data lists of poland (wo j e w o d a , ł a w r y n o w i c z 1 9 9 2 ) or of germany (b e n k e r t et al. 1996). in regard to host selection its interrelationship to genera and species of bryophytes (see 5.71 / 5.72) appears not very much specialized. the occurrence on few genera of bryophytes only would result in a higher risk for its existence. according to our present knowledge r. swartzii does not occur on sphagnum and not on foliose or thallose liverworts. the host selection is somewhat narrower than in case of r. fibula, however, much broader than in case of loreleia-species. 200 a. bresinsky and a. schötz rickenella fibula (bull.: fr.) raithelh. syn.: gerronema fibula (bull.: fr.) singer, rickenella aulacomniophila kost observations. mycelial cultures produce in accordance with the colour of the fruit bodies an orange pigment, at least under lucky circumstances. in all other cultural characters r. fibula is similar to r. swartzii. in addition to the observations of l a m o u r e (1979) on dispersed growing colonies some microscopic features could be observed which are characteristically for colonies of rickenella growing in more or less dense clusters. in such a situation the dicaryotic hyphae are composed of rather short cells and clamps are often missing at the septa (fig. 2a). hyphal branches show often chains of chlamydospores. these germinate sometimes in situ with clamped hyphae (fig. 2e). it seems that chlamydospore formation is induced by the existence of many zones of inhibition created by dense growth of numerous colonies on restricted space. in contrast to this crowded situation, typical chlamydospores are not developed from isolated growing mycelia. many weeks later, peziza-like structures occasionally appeared between the densely growing dicaryotic colonies. these peziza-like structures are in terms of their hyphae similar to dicaryotic hyphae, however, clamp-less and with uninucleate instead of binucleate cells (fig 2f). in freshly isolated cultures the mycelial growth exceeded with diameters of colonies up to 25 mm (in our experiments) that of older strains with diameters between 4 and 16 mm after 6 weeks (l a m o u r e 1979). the cellulase activity appeared in rickenella to be lower than in l. marchantiae and l. postii [and of course lower than in the wood inhabiting species gerronema (chrysobostrychodes) chrysophyllum and g. strombodes]. this observation is in agreement with the assumption that rickenella-species do not cause any damage to their bryophyte hosts by virulent attack of the cell walls of living cells. rickenella-species are not strong parasites rather than either saprobionts (living from the death parts of their hosts which in this case are not severely influenced by the hosted fungus) or even symbionts (establishing some kind of endomycorrhizal interrelationship as assumed by k o s t 1984). the latter assumption is supported by field observations: rickenella fruited year by year on the same bryophyte thallus without setting a remarkable harm to its host. fruiting began according to our observations on hylocomium splendens in the mid of august when fresh green branches arised from the cauloids of the preceding year; then the fruit bodies of rickenella fibula were fixed just underneath the fresh green sections on those parts of the cauloids which were bearing dead leaflets only. eco-geogramm for rickenella fibula (using database pilzoek): sources: achhammer 1986; albers, grauwinkel 2005; antonin, noordeloos 2004; arnolds 1977, 1981; bas et al. 1985; besl et al. 1982; bresinsky 1997a, 1998, 1999, 2000, 2001; bresinsky, einhellinger 1987; bujakiewicz 1973, 1979,1986, 1993; bujakiewicz, fiebich 1991/92; courtecuisse 1994; dehnert 2002; derbsch, schmitt 1987; dyląg, gumińska 1997; einhellinger 1973, 1976, 1977, 1981, 1982; favre 1955; garrido 1985; grauwinkel 1987; gulden, torkelsen 1996; gumińska 1976, 1992; haeupler et al. 1981; hallgrimsson 1981; hansen, knudsen 1992; hauck 1993; helfer 2001; imazeki, hongo 1984; jahn et al. 1967; jelik, tortic 1973; kalamees, vaasma 1981; karasch 2001; kost 1984; kreisel 1957, 1970, 1987; krieglsteiner 1977, 2000; krieglsteiner l. 1999, 2001; krieglsteiner,. luschka 2000; krisai 1987; kühner, lamoure 1986; kummer 1997; lange 1957; ławrynowicz, szkodzik 1998; legon, henrici 2005; lisiewska behavior in cultures and habitat 201 fig. 2: rickenella fibula in mycelial culture: a) dicaryotic mycelium; some clamps are visible at the septa of hyphae. in some hyphae a transition to short, rather broad and clamp less hyphae occurred; some of them form thick walled elliptic chlamydospores (chl); b) dicaryotic, clamped hyphae (cl); some of them inflated and vacuolized. terminal elements sometimes similar to cystidia (cy); c) monocaryotic hyphal segments formed by germinating chlamydospores (chl); d) dicaryotic hy phae formed by germinating chlamydospores; e) chains of chlamydospores partially germinating with dicaryotic hyphae; f) chlamydospores and their hyphal outgrowths; taken from a culture with peziza like structures. 202 a. bresinsky and a. schötz 1987; lisiewska, reszel 2000; luschka 1993; łuszczyński 1998; malençon, bertault 1975; miller, farr 1975; möller 1945; monthoux, röllin 1993; neuhoff 1949; nuss 1999; oertel, fuchs 2001; paulus 1991; pearson 1950; ricek 1989; rimóczi 1994; rücker 1990; rücker, wittmann and peer 1989; sedlmeir 1985; schmid-heckel 1985, 1988, 1989; senn-irlet 1987c; skirgiełło 1998; šmarda 1972, 1973; sonneborn, stangl 1970; stangl, sedlmeir and geh 1987; stasińska, sotek 2004; tejera 1980; urbonas, kalamees and lukin 1986; vaasma, kalamees and raitviir 1986; watling, gregory 1977; watling, rotheroe 1989; winterhoff 1980, 1993, 1993b, 1994, 1995, 2001, 2002; wojewoda 2003; wöldecke kl. 2001; wöldecke 1990, 1998; wöldecke, wöldecke 1988, 1992; zehfuß, ostrow 2004. for references see http://www.pilzoek.de a. nutrition: 0.1 bryicolous parasite (?), 0.32 bryicolous endotrophic mycorrhiza (?), 0.5 bryicolous saprobiont (?). b. substrates: 2.12 bryophyte covered wood of conifers: dead. 2.32 bryophyte covered wood on soil surface (alnus). < 2.6 carbophilous, on or near charcoal > [pl]. 2.711 standing stem wood. 2.72 fallen stem wood, fallen knot wood. 3.4 between litter from grasses and herbals [a]. 5.0 interrelationship with bryophytes as substrates: < 5.1 liverworts >, 5.2 mosses, 5.3 peat mosses. 5.71/5.72 (connected to living bryophytes): atrichum undulatum, aulacomnium palustre, barbula convoluta, brachythecium albicans, brachythecium rutabulum, bryum capillare, bryum pseudotriquetrum, calliergonella cuspidata, campylopus, ceratodon purpureus, cirriphyllum piliferum, climacium dendroides, dicranella heteromalla, dicranoweisia cirrata, dicranum bonjeanii, dicranum majus, dicranum scoparium, distichum capillaceum [pl], eurhynchium angustirete, eurhynchium praelongum, eurhynchium striatum, homalothecium lutescens, hylocomium splendens, hypnum cupressiforme, hypnum cupressiforme var. lacunosum, hypnum jutlandicum, hypnum lindbergii, leucobryum glaucum, < lophocolea bidentata > ,< ? marchantia polymorpha [pl]; cf. loreleia marchantiae as possible fungus in this context >, mnium hornum, philonotis fontana [pl], plagiomnium affine, plagiomnium undulatum, pleurozium schreberi, pogonatum urnigerum, pohlia nutans, polytrichum formosum, polytrichum juniperinum, polytrichum piliferum,< ptilidium ciliare >, rhytidiadelphus squarrosus, rhytidiadelphus triquetrus, rhytidium rugosum, scleropodium purum, sphagnum compactum, sphagnum contortum, sphagnum inundatum, sphagnum nemoreum, sphagnum papillosum, sphagnum rubellum,sphagnum warnstorfii, thuidium tamariscinum. c. habitats: woods / copses: 10.0 beech woods, mixed beech forests, (fagion sylvaticae pr. m. p.): 10.1 beech woods on (moderately) rich brown earth: 10.11 galio odorati-fagenion: 10.11.01 galio odorati-fagetum, 10.11.02 hordelymo-fagetum. 10.12 beech woods with share of white wood fir (abies; lonicero alpigenae-fagenion): 10.12.01 dentario enneaphylli-fagetum [tch, pl], 10.12.04 lonicero alpigenae-fagetum. 10.2 beech woods on rendzina and pararendzina soils (cephalanthero-fagenion): 10.2.01 carici-fagetum. 10.3 beech woods and mixed woods with oak and beech on acid soils (luzulo-fagenion): 10.3.01 luzulo-fagetum, 10.3.02 luzula pilosa-fagus sylvatica-community [pl]. 11.0 mixed frondose woods without (dominating) beech: 11.1 humid woods with maple (aceri-fagenion): 11.1.04 fraxino-aceretum pseudoplatani. 11.2 mixed lime-tree woods [pl], 11.3 oak-hornbeam woods (carpinion betuli): 11.3.01 galio sylvatici-carpinetum, 11.3.02 stellario holosteae-carpinetum betuli. 11.4 termophilic mixed oak woods (quercetalia pubescentis; quercion pubescenti-petraeae [h], potentillo albae-quercion petraeae): 11.4.02 quercetum pubescenti-petraeae [h], 11.4.05 behavior in cultures and habitat 203 potentillo albae-quercetum petraeae. 11.5 mixed oak woods on acid soils (quercenion robori-petraeae [pl], genisto tinctoriae-quercenion robori-petraeae): 11.5.01 holco mollis-quercetum, 11.5.02 betulo-quercetum petraeae, 11.5.03 genisto tinctoriae-quercetum petraeae. 12.0 conifer dominated woods (natural): 12.1 woods with high share of silver fir (vaccinio-abietenion): 12.11.01 galio-abietetum [pl], 12.12.01 vaccinioabietetum. 12.2 spruce woods outside bogs: 12.2.01 bazzanio-piceetum, 12.2.02 calamagrostio villosae-piceetum. 12.5 scots pine woods outside bogs: 12.51.02 moliniopinetum, 12.51.03 calamagrostio-pinetum, 12.51.06 pyrolo-pinetum sylvestris . 12.52 dicrano-pinion: 12.52.01 leucobryo-pinetum. 13.0 ombrogenous bogs and associated plant communities (oxycocco-sphagnetea; rhynchosporion albae): 13.1.03 sphagnum cuspidatum-eriophorum angustifolium-community [pl]. 13.2.03 sphagnetum magellanici, 13.2.04 eriophoro-trichophoretum cespitosi. 13.32 woods on bogs with scots pine: [pl]: 13.32.01 vaccinio uliginosi-pinetum sylvestris. 13.33 woods on bogs with spruce. 13.34 woods on bogs with birch [pl]: 13.34.01 vaccinio uliginosi-betuletum pubescentis, 13.34.03 betula pubescens-sorbus aucuparia-community. 14.0 woods of river lowlands and swamps: 14.1 flood-plain woods: 14.11 salicion elaeagni: 14.11.03 salici-hippophaëtum rhamnoidis [ch]. 14.14 mixed woods with ash and alder along brooks: 14.14.03 carici remotae-fraxinetum, 14.14.04 stellario nemorum-alnetum glutinosae, 14.14.06 pruno-fraxinetum. 14.15 hard-wood floot plain forests: 14.15.01 querco-ulmetum minoris. 14.2 paludal forests: 14.21 alder swamp forests (alnenion glutinoso-incanae, alnion glutinosae): 14.21.01 carici elongatae-alnetum glutinosae, 14.21.03 caltha palustris-alnus glutinosa-community. 14.22 downy birch swamps and grey willow swamps: 14.22.03 salicetum auritae, 14.22.04 salicetum cinereae. 15.0 mantles of woods / bushes / hedges incl. herbal fringes: 15.22.07 urtico-aegopodium podagrarariae. 16.0 man made forests / non-natural wood communities: 16.1 coniferous forests: 16.11 pine forests: 16.11.01 pure scots pine forests: 16.11.05 scots pine forests with empertum [pl]. 16.111.15 scots pine forests on lime stone soils. 16.112 plantations of austrian pine (pinus nigra). 16.12 spruce forests: 16.12.01 typical oxalis-spruce forests [tch], 16.12.12 nude spruce forests without herbaceous layer [tch], 16.12.25 spruce forests on basic (lime stone) soils. 16.14 larch forest plantations. 16.2 deciduous forests: 16.24 birch forests, 16.25 robinia forests, 16.26 poplar forests / plantations, 16.27 plantations of willows. 18.0 open areas within or near woods: 18.1 forest clearances: 18.12.01 rubetum idaei. 18.2 margins of forests [a], 18.3 paths through forests; 18.31.16 calluno-sarothamnetum. 19.4 garden copses. outside woods and copses: 21.0 paludal areas around springs / vegetation around headwaters (cratoneurion commutati): 21.2 tuff beds. 22.0 soligenous bogs / transitional bogs (scheuchzerio-caricetea fuscae, caricion davallianae): 22.1.01 orchioschoenetum nigricantis, 22.1.02 primulo-schoenetum ferruginei. 22.2 acidophilous, non-calcareous reed marshes with dwarf sedges (juncion acutiflori): 22.2.05 juncetum acutiflori [nl]. 22.3 transitional bogs [a]: 22.3.01 caricetum lasiocarpae [a, pl]. 24.0 dunes / dry grassland on sand: 24.12 ammophiletae arenariae.24.13 grey dunes, 24.15 bush communities on dunes. 24.2 inland dunes and associated grassland communities (thero-airion, corynephorion canescentis): 24.2.01 spergulo vernalis-corynephoretum canescentis, 24.2.05 airo caryophylleae-festucetum ovinae [nl], 24.2.06 thymo-festucetum ovinae [nl], 24.2.13 diantho deltoidis-armerietum elongatae, 24.2.14 armerio-festucetum trachyphyllae. 25.4 snow pockets: 25.42.01 salicetum herbaceae. 25.54 calcareous fens (caricion davallianae). 27.0 grasslands / drifts: ses204 a. bresinsky and a. schötz lerion albicantis [pl]. 27.1 dry and medium dry grasslands: 27.11 continental steppe grasslands (festucion valesiacae): 27.11.04 adonido-brachypodietum pinnati. 27.12 submediterranean steppe grassland (mesobromion, xerobromion): 27.12.03 gentianokoelerietum, 27.12.06 viscario-festucetum heteropachyos, 27.12.09 trinio-caricetum humilis, 27.12.10 pulsatillo-caricetum humilis. 27.2 dwarf shrub heathes, drifts, mattgrass areas on acid soils (genistion pilosae, nardion): 27.21.02 genisto pilosae-callunetum [nl], 27.21.09 aveno-genistetum sagittalis. 27.23 matt-grass areas on lower (colline) elevations (violion, juncion squarrosi): 27.23.01 polygalo-nardetum, 27.23.03 juncetum squarrosi [nl]. 28.0 meadows / pastures: 28.1 rich meadows / pastures / cultivated grassland (arrhenatherion elatioris): 28.11.01 arrhenatheretum elatioris. 28.12.02 geranio-trisetetum. 28.13 rich pastures / trampling resistant grassland: 28.13.03 festuco-cynosuretum, 28.13.04 lolio-cynosuretum, 28.13.14 prunella vulgarisplantago major-community. 28.14 oligotrophic grassland. 28.16 garden lawns. 28.2 moist and wet meadows (calthion): 28.21.01 juncetum subnodulosi, 28.21.02 angelicocirsietum oleracei, 28.21.06 scirpetum sylvatici [nl], 28.21.08 epilobio-juncetum effusi. 28.22 litter meadows / meadows under condition of changing moisture (molinion caeruleae, cnidion dubii): 28.22.01 molinietum caeruleae, 28.22.03 cirsio tuberosi-molinietum arundinaceae [nl]. 29.0 river marshes / river banks / alluvial meadows: 29.21 reeds (magnocaricion, caricion elatae): 29.21.18 caricetum paniculatae, 29.21.19 caricetum rostratae [a]. 29.22 filipendulion: 29.22.02 filipendulo-geranietum palustris. 30.0 cultivated land / waste land / depositions: 30.7 gardens. 31.1 ruderal sites / weed vegetation, 31.4 slag-dumbs / coal-dumbs, 31.7 pits (sand-. loam-, clay-), < 31.8 outdoor fire places [pl]; cf. loreleia postii; l. marchantiae >. 32.3 field-paths / meadow-paths: plantaginetea majoris [nl], agropyretea intermedio-repentis. fungal communities: 38.42.11 boleto (aerei)-russuletum luteotactae [tch]. 38.42.41 geastro (quadrifidi)-agaricetum semotae [tch], 38.42.42 clitocybe (brumali)-phellodonetum nigri [tch] d. soil characters: 40.1.1 basalt [tch], 40.3 loess [tch], 40.4 sand, 40.8 coal / coal stacks: 40.8.1 brown coal [pl]. 41.1 mull, 41.5 peat. 42.1 calcareous soils; 42.22 sand soils, siliceous sands; 42.3 brown soils, 42.61 alluvial soils. 43.0 acidity of soils: 43.1 acid, ph 3.0-5.6; 43.1.2 ph 3.4-4.0 [tch]; 43.1.3 acid, ph 4.1-4.8; 43.1.5 moderately acid, ph 4.9-5.6 [pl]. 43.2 neutral, ph 5.7-6.5 [tch]: 43.2.7 weakly acid / weakly basic, ph 5.7-6.5 [h]. 43.3 basic, ph 6.6-7.5 and higher [tch]. – 44.2 moderately rich on nutrients, 44.3 rich on nutrients (nitrogen). 45.1 dry; 45.2 humid, mesic; 45.3 wet. f. temperature and light at the stand: 50.2.3 cool, 50.4.7 warm. 51.1 shadow [a], 51.7 half light, 51.8 light. g. phenology, persistence and sociability of fruit bodies: < 52.1 spring >, 52.2 summer, 52.3 autumn, < 52.4 winter >. 53.2 short living fruit bodies. 54.1 single, 54.2 in groups, 54.3 in herds. i. distribution: 59.11 europe, north: isl, far, spi, n, dk, s, sf, lit, lat, est, rus. west: gb, f, b, nl. central: ch, d, a, tch, pl, h, dk. east: h, ukr, rus, lit, lat. south: p, sd, i, sc, cro, tq. 59.12 asia: caucasus, kamchatka, japan. 59.13 north america. 59.15 north africa: marroco, tunisia. 59.16 oceanic islands north of the equator: canaries, greenland. 59.2 southern hemisphere: 59.22 south america, outside tropics: chile; 59.23 australia, new zealand? 59.3 tropics (south america, venezuela). behavior in cultures and habitat 205 k. distribution and endangerment in central europe: 61.3 alpid, 61.4 arcticalpid, 61.5 temperate-central european, 61.6 boreal, 61.7 arctic.62.1 planar, 62.2 colline incl. submontane (330-350 m), 62.3 montane (575-650 m), 62.4 high-montane, 62.5 subalpine, 62.6 alpine (ch, f: 2500 m; pl: 1340-1520 m). – 65.0: endangerment: not threatened. comments. fruit bodies of rickenella fibula appear on all major groups of mosses including peat mosses (sphagnum) and even few representatives of foliose hepaticae (plagiochila, ptilidium). on the latter group of bryophytes the fungus occurs only exceptionally and it might remain doubtful if foliose hepaticae really are taken as a substratum or if the mycelium of the fungus is attached to them loosely and then by chance only. besides many species and genera of mosses also the liverwort marchantia polymorpha has been listed once to be associated with rickenella fibula (“among specimens of marchantia polymorpha”; d y l ą g , g u m i ń s k a 1997). it is uncertain if in this case marchantia really served as substrate. especially if the fungus is found on burnt places (31.8) in association with marchantia, it is not unlikely that rickenella fibula might be mistaken for loreleia marchantiae. attention should also be paid to r. pseudogrisella (a.h. smith) gulden. this species is bound to the liverwort blasia pusilla and was found in arctic-montane habitats of scandinavia (n, s, sf), of island and of greenland (h a n s e n , k n u d s e n 1992; l u d w i g 2001); it has not been recorded so far from poland neither from germany where it could, however, be expected in the alpine zone. records resembling r. fibula, but exhibiting more dull ochraceous instead of vivid orange colours of fruit bodies, should be identified as r. mellea (singer et clémençon) lamoure. this taxon has been observed among mosses of the genera bryum and philonotis; it has been found recently in germany (l u d w i g 2001) and may occur in poland too. attempts to distinguish other taxa on the species level bound to special host species of bryophytes (f. e. rickenella aulacomniophila kost on aulacomnium palustre) failed. the discriminating characters like different size and shape of cystidia turned out to be extremely variable within a rather broad range of continuous variation; this might also apply on differences in spore sizes in r. mellea as compared to r. fibula. in bavaria the highest frequency of fruit bodies of rickenella fibula had been observed by us 1985 on hypnum cupressiforme, dicranella heteromalla, rhytidiadelphus squarrosus, brachythecium rutabulum, plagiomnium affine, hylocomium splendens and pleurozium schreberi. on a single patch of moss, measuring approx. 10 cm in diameter, 10 or even more fruit bodies have been counted in case of hypnum cupressiforme, dicranella heteromalla or plagiomnium affine. in different months of the season different species of bryophytes were preferably carrying fruit bodies. in the area of regensburg, bavaria (125 records evaluated), hylocomium splendens was observed to bear fruit bodies mainly in august, plagiomnium affine and pleurozium schreberi mainly in september and dicranella heteromalla mainly in october at the end of the mushroom season (then being the most important host to carry carpophores of the fungus). the fungus can be attached to different parts of the host: main axis and lateral branches of cauloids, rhizoids and leaflets. in central and western european countries rickenella fibula shows a high overall frequency of occurrence. it is far from being endangered because of its adaptation to a great variety of quite different habitats [dry (27.11, 27.12) or wet (13.0, 14.2, 206 a. bresinsky and a. schötz 21.0, 22.0), poor or rich in nutrients (24.2, 27.2 versus 28.1, 30.7, 31.1, 32.3) on acid or on neutral or even basic soils (27.23 versus 27.11), in man made sites (31.4, 31.7), etc.] and because of its association with numerous bryophytes serving as substrates and growing under many different circumstances (5.71 / 5.72: f. e. the ubiquitous and nitrotolerant rhytidiadelphus squarrosus). discussion the species of the genera loreleia and rickenella are not regarded to be endangered in general. this applies to some extent even on species which are quite rare or only occasionally observed. rickenella swartzii with a low grade of frequency as compared to r. fibula is nevertheless not threatened. its low frequency of records may be caused by a lower rate of readiness for fruiting. this does not necessarily mean endangerment since a high potential is inherent in rickenella (and loreleia) to disperse by asexual germ cells (oidia, chlamydospores). the habitat requirements are broad enough to include sites with higher contents of nutrients in the soil (28.11.01, 28.13, 30.7, 30.91), and the association to bryophytes as substrates is wide enough to permit the fungus to be present on bryophytes which are not sensitive against man made environmental changes and restrictions (5.71 / 5.72: f. e. hypnum cupressiforme, rhytidiadelphus squarrosus). the estimated rarity of the loreleia species may be the result of inadequate observation. in case of l. marchantiae the host plants, mats of thallose liverworts, are not checked intensively and regular enough by mycologists over a greater period throughout the whole year. thus the fructification of l. marchantiae, if it occurs for a short time, may be overlooked quite often. one has to consider that the fungus might be well distributed in its mycelial stage. it is, however, seemingly rare since observations are based on fruit bodies only. it is hard to believe that a fungus which is easily growing in cultures and which is provided by different, readily germinating propagules should not be fit enough to find spontaneously its hosts in order to develop mycelia and asexual diaspores. the low range of hosts and the specialization on genera with only few species (as in marchantia and in lunularia) does apparently not afford a higher rate of genetic variation which would be warranted by sexual reproduction in fruiting bodies. once being adapted to its few (in terms of evolution more or less stabilized) hosts, such fungal species may be not dependent on sexual reproduction in fruit bodies for their survival. this might be one reason for rare fructification. again the rating of being not endangered is based on the evaluation of habitats and substrates. these (i. e. the substrates for loreleia marchantiae) are not recorded to decrease or being threatened; in other words, the species within marchantia and lunularia, serving as substrates, are not listed as being endangered. lunularia cruciata is quite often found in man made habitats like parks, green houses, flower pods etc. it shows an increasing tendency to expand and to invade also natural habitats in central europe. it is, however, susceptible to freezing temperatures and may die back in a cold winter; its distribution is much more oceanic than continental. within marchantia polymorpha it is ssp. ruderalis which is the most distributed infraspecific taxon, whereas ssp. polymorpha is more restricted in its distribution. it is not likely that loreleia marchantiae does prefer one of the both subspecies as a host. behavior in cultures and habitat 207 wide availability of suitable substrates is warranted also in this case of substrate interrelationship, because of a sufficient wide range of appropriate habitats and because of the synanthropic and nitrotolerant character of the host species (especially marchantia polymorpha ssp. ruderalis). the evaluation of endangerment in case of l. marchantiae gets a new dimension in the light of a possible interrelationship with two partners. the dependence from cyanobacteria in connection with liverworts complicates the existence of the fungus in a sense that the chain of nutrition is not easily established and if so, then it is more vulnerable. such complex interdependence causes a higher degree of possible endangerment. finally it has to be taken into account that unclear taxonomic concepts on the species level complicate the evaluation of the endangerment of species. a narrow species concept within rickenella (fibula) would distinguish separate micro-species on special bryophytes serving as substrates (f. e. r. aulacomniophila on aulacomnium). some of the taxa to be distinguished in a narrower sense could fulfil the criteria of being threatened because of their more restricted range of substrates and habitat conditions. the transfer of r. aulacomniophila into the synonymy of r. fibula, as it has been advocated in this paper, includes the risk of getting sightless against the situation of actually threatened organisms. in the context of loreleia marchantiae the risk of endangerment would be lower if this species would fall into synonymy with l. postii. in this case the range of substrates, including mosses and thallose liverworts as well, and the habitat requirements would be quite wider than it is supposed in the view of a narrow species concept. references a c h h a m m e r ( s c h ö t z ) a. 1986. kulturmerkmale und ökologie einiger omphaloider tricholo mataceen (gattungen chrysomphalina, gerronema, rickenella). ls für botanik, univ. regensburg (msc.). a n d e r s o n j.j.w. 1962. studies on micrococci isolated from the north sea. j. appl. bact. 25: 362 368. b a s c., k u y p e r t h . w., n o o r d e l o o s m.e., ve l l i n g a e.c. 1995. flora agaricina neerlandica 3. rotterdam. b e n k e r t d. et al. 1996. rote liste der großpilze deutschlands. schr. r. f. vegetationskunde 28: 377 426. b o i d i n j. 1951: recherche de la tyrosinase et de la laccase chez les basidiomycetes en culture pure. revue mycol. 16: 173. b r e s i n s k y a., d ü r i n g ch. 2001. pilzoek, ein erfassungsprogramm für daten zur ökologie und chorologie von pilzen in mitteleuropa. z. mykol. 67: 157 168. b r e s i n s k y a., d ü r i n g ch., a h l m e r w. 2005. datenbank pilzoek jetzt über internet zugänglich (http://www.pilzoek.de). z. mykol. 71: 201 209. b r e s i n s k y a., k r e i s e l h., p r i m a s a. 1995. mykologische standortkunde. regensburger mykol. schr. 5: 1 304. c l é m e n ç o n h. 1982. kompendium der blätterpilze. europäische omphalinoide tricholomataceae. z. mykol. 48: 195 237. d e r b s c h h. 1977. seltene agaricales aus dem saarland 16: gerronema postii. z. pilzk. 43: 180 181. d y l ą g e., g u m i ń s k a b. 1997. postfire macromycetes from deciduous wood in the chrzanów forest inspectorate (s poland). acta mycol. 32: 173 187. g u m i ń s k a b., m i e r z e ń s k a m . 1990: gerronema marchantiae sing. et clem. a fungus associated with marchantia polymorpha l. and nostoc sp. zesz. nauk. uniw. jagiell. prace bot. 24: 171 177. h a n s e n l., k n u d s e n h. 1992. nordic macromycetes 2. copenhagen. h o r a k e. 2005. röhrlinge und blätterpilze in europa. spektrum. heidelberg. 208 a. bresinsky and a. schötz k l á n j. 1992. two gerronema species growing on marchantia thalli. 1. gerronema marchantiae sing. et clém., 2. g. postii (fr.) sing. česká mykol. 46: 121 125. k o s t g. 1984. moosbewohnende basidiomyceten 1: morphologie, anatomie und ökologie von arten der gattung rickenella raithelh.: rickenella fibula (bull.: fr.) raithelh., r. aulacomniophila nov. spec., r. swartzii (fr.: fr.) kuyp. z. mycol. 50: 215 240. l a m o u r e d. 1979. charactères morphologiques, caryologiques et culturaux des mycéliums de trois espèces de rickenella (agaricales). sydowia beih. 8. ann. mycol. ser. ii: 251 254. l i s i e w s k a m., b a l c e r k i e w i c z s. 1991. macrofungi in orchards treated with herbicides. boletus 15 (2): 45 56. l u d w i g e. 2001. pilzkompendium. eching. l y r h . 1958: über den nachweis von oxydasen und peroxydasen bei höheren pilzen und die bedeutung dieser enzyme für die bavendamm reaktion. planta 50: 359 370. ł a w r y n o w i c z m., b u j a k i e w i c z a., m u ł e n k o w. 2004. mycocoenological studies in poland (1952 2004). monogr. bot. 93: 1 102. m o s e r m. 1960. die gattung phlegmacium. bad heilbrunn. n e s p i a k a.: see references in ł a w r y n o w i c z et al. (2004). n i c o l a s g. 1967. association des bryophytes avec d’autres organismes. (in:) ve r d o o r n (ed.). manual of bryology. amsterdam. p r i l l i n g e r h., s i x w. 1983. genetische untersuchungen zur fruchtkörper und artbildung bei ba sidiomyceten: genetische kontrolle der fruchtkörperbildung bei polyporus ciliatus. pl. syst. evol. 141: 341 371. s c h m i d h e c k e l h. 1985. zur kenntnis der pilze in den nördlichen kalkalpen. nationalpark berch tesgaden. forschungsberichte 8: 1 201. s i n g e r r. 1986. agaricales in modern taxonomy. 4th ed. koenigstein. s k i r g i e ł ł o a.: see references in ł a w r y n o w i c z et al. (2004). ta n s e y m. r. 1971. agar diffusion assay of cellulolytic ability of thermophilic fungi. arch. microbiol. 77: 1 11. we y l a n d h. 1970. zur isolierung und identifizierung mariner bakterien. verh. inst. meeresforsch. bremerhaven 12: 289 296 w o j e w o d a w. 2003. checklist of polish larger basidiomycetes. (in:) z. m i r e k (ed.). biodiversity of poland 7. w. szafer institute polish academy of sciences, kraków. w o j e w o d a w., ł a w r y n o w i c z m. 1992. red list of threatened macrofungi in poland. (in:) k. z a r z y c k i , w. w o j e w o d a , z. h e i n r i c h (eds). list of threatened plants in poland. 2 ed.: 27 56. w. szafer institute of botany, polish academy of sciences, kraków. rozwój w kulturach i wymagania siedliskowe gatunków z rodajów loreleia i rickenella (agaricales) s t r e s z c z e n i e termin eko geogram został wprowadzony dla uwidocznienia danych ekologicznych i geo graficznych w powiązaniu z poszczególnymi gatunkami grzybów. w tym celu opracowano bazę danych pilzoek. w pracy zaprezentowane są przykłady możliwosci odtworzenia eko geogramów za pomocą bazy danych pilzoek dla agarikoidalnych grzybów: loreleia marchantiae, l. postii, rickenel la swartzii i r. fibula. w odniesieniu do wymagań siedliskowych dyskutowany jest stopień po tencjalnego zagrożenia gatunku. europejskich gatunków loreleia nie uważa się za zagrożone w środkowej europie, jednak istnieje ryzyko pojawienia się zagrożenia z uwagi na niski stopień frekwencji owocników i wąską specjalizację w odniesieniu do substratu. kultury rickenella fi bula, r. swartzii, loreleia marchantiae (= gerronema daamsii) and l. postii były badane pod względem akumulacji pigmentu, tworzenia się chlamydospor i niektórych innych cech. 2014-01-01t11:44:14+0100 polish botanical society lichens of neglected habitats in eastern and east-central european lowlands jurga motiejūnaitė laboratory of mycology, institute of botany, žaliųjų ežerų str. 49, lt 08406 vilnius 21, mikojm@botanika.lt m o t i e j ū n a i t ė j . : lichens of neglected habitats in eastern and east central european lowlands. acta mycol. 41 (1): 145 154, 2006. situation of lichens of aquatic and transient habitats in eastern and east-central european lowlands is discussed basing on example of several selected species: leptogium biatorinum, sarcosagium campestre, steinia geophana, verrucaria aquatilis, v. hydrela, v. praetermissa, v. xyloxena. both habitat types are generally very much neglected in the region and all species show large spatial gaps in recording, which makes it difficult to judge both about their true distribution limits and spreading dynamics. on the other hand, targeted search through the suitable habitats and abundance of such indicate that many of these lichens are probably not uncommon in the region. key words: lichens, biogeography, aquatic habitats, transient habitats introduction rare lichens often make significant parts of national floras and provide the basis for red lists. however, “rare” might comprise both declining species and stable or spreading ones. declining species are usually well known, more or less historically documented and confined to the habitats and substrates that are traditionally investigated by lichenologists. meanwhile situation of many species defined as “rare” in national floras often remain obscure. especially this is true for eastern and eastcentral (eec) europe. the reasons for some lichen to be defined as rare can be several: 1. the species are “new” – relatively recently described. 2. the species are taxonomically “difficult”, they are often reported as complexes or not separated from similar species. 3. the species are connected to particular habitats or niches that are neglected by lichenologists. acta mycologica vol. 41 (1): 145-154 2006 it is a great pleasure and honour to dedicate this paper to professor alina skirgiełło, who always cared for lithuania and its mycology 146 j. motiejūnaitė “new”, recently described species comprise lichens, which have been described during the last two decades. presence of such species in the national lists generally indicates degree of recent lichenological activity (see e.g. m o t i e j ū n a i t ė 2005). taxonomically „difficult“ species are either scarce or totally excluded from the floras or inventory lists of eastern europe (e.g., most of sterile crustose lichens) or poorly distinguished, e.g. see notes for acrocordia cavata in m o t i e j ū n a i t ė (2005) or anisomeridium polypori in c z y ż e w s k a et al. (2005). species associated with habitats neglected by lichenologists make a weird group, indicating, to certain extent, activities of lichenologists, the variability of fieldwork and nature of lichenological coverage. species of this group are often mentioned in discussions about lichen flora dynamics, as most of them are considered to be rare in many european countries; a sudden boost in locality numbers after specific habitat studies might result in a discussion as to whether the species is spreading or not, whether the species is a neophyte or a long-present member of the national flora. typical examples of neglected habitats are lowland freshwater bodies and transient habitats, both of which receive only sporadic attention from lichenologists. the purpose of this paper was to review situation of lichens associated with both these habitats in eec europe and to attract attention to the importance of study of these habitats. for the aims of the paper several species should be characteristic to aquatic and transient habitats were selected. their known distribution and situation in eec europe were analysed mainly on the basis of all available literature sources and to some extent of herbarium collections (mainly lithuanian). territory under discussion for the review temperate and cool temperate forest zone of lowland eec europe (between ca. 20o and 55o longitude and 53o and 63o latitude) was chosen. though the area is biogeographically complex, it still holds certain homogeneity pertaining conditions for lichen distribution. there are no large natural obstacles for lichen dispersal in this area, the only limiting factors for their spread being climatic and edaphic. though land-use history and intensity vary from country to country, there are still many similarities connected with long periods of similar administrative history. many traditional rural activities persist, such as use of local building materials resulting in sand-, graveland loam-pits, and of untreated timber for buildings (although the latter tendency is decreasing lately at least in part of the territory). administratively the territory includes polish lowlands (p), baltic countries – lithuania (li), latvia (la), estonia (e), belarus, and the forest zones of the ukrainian lowlands (u) and of european russia (r). results and discussion lichens of aquatic habitats. aquatic habitats, though locally well studied in western and central europe (see e.g. g i l b e r t 1 9 9 6 ; g i l b e r t , g i a v a r i n i 1 9 9 7 ; m o l i t o r , d i e d e r i c h 1 9 9 7 ; k e l l e r 2 0 0 0 ; t h ü s 2 0 0 2 ) are very much under-recorded in eec europe, especially in its lowland parts. both numbers of recorded species are low and the records themselves are sparse, though when well studied, aquatic environs show to be rich in lichens. crustose lichens are espe lichens of neglected habitats 147 cially understudied, among which perithecioid species prevail. verrucaria aquatilis, v. hydrela and v. praetermissa are the most common freshwater aquatic perithecioid lichens in the region, their distribution and status reflects situation of whole group. distribution of these lichens in eec europe reflects solely regional activity of lichenologists and gives no clear pattern of geographical ranges. most of them (except v. hydrela) are attributed to suboceanic species, occurring mainly in temperate zone. generally, these lichens are quite well known in western and central europe, but are very much under-recorded in the east. aquatic verrucaria species are found almost exclusively in southeastern and eastern region of the baltic sea (figs. 1-3). though there is no shortage of suitable habitats in eastern europe, aquatic lichens, especially crustose species are almost utterly omitted from lichen inventories. judging by the known distribution and tendencies of recording, aquatic verrucaria species should be found also further eastwards in pristine streams of forested areas, as it is demonstrated by a record of v. hydrela outside of the discussed area, in southern part of the baikal lake region (u r b a n a v i c h e n e , u r b a n a v i c h u s 1999). besides, these lichens are source of taxonomic confusion, which is discernible even among scarse records (e.g., the record of v. praetermissa in kaliningrad region, russia (a n d r e e v 2002) suggests v. elaeina (see o r a n g e 2000). frequencies of these lichens in the discussed region are as hypothethical, as their distribution ranges. in most cases they range from very rare to rather rare in all countries (tab. 1), though it is admitted that overlooking is the main reason for the record scarcity (e.g. for verrucaria hydrela in northern poland (j a n d o , k u k w a 2003). dynamics of these species as well as conservational value are even less clear. generally, aquatic lichens are connected with vulnerable habitats, as most of them inhabit pristine streams, little influenced by eutrophication (see g i l b e r t 1996; g i l b e r t , g i a v a r i n i 1997; t h ü s 2002; m o t i e j ū n a i t ė 2003). this is well expressed by the fact that aquatic verrucaria species are red-listed in west-european areas of intensive urban and agricultural land use: v. aquatilis and v. hydrela – in denmark (s ø c h t i n g 1997) and lowland lands in germany (variable categories, w i r t h et al. 1996); v. praetermissa – in german lowland lands (critically endangered to rare). thus, they could be attributed to habitat-demanding and decreasing species. in eec europe, aquatic verrucaria species are red-listed only in poland (tab. 1). fig. 1. distribution of verrucaria aquatilis in eastern and east central europe � records more than 50 year old, � modern records fig. 2. distribution of verrucaria hydrela in eastern and east central europe � records more than 50 year old, � modern records 148 j. motiejūnaitė fig. 3. distribution of verrucaria praetermissa in eastern and east central europe � modern records lichens of transient habitats. lichens of transient habitats are ephemeral, generally inconspicuous and rarely recorded, therefore are presumed to be nationally rare in almost all european countries. most often such habitats are man-made, like bare soil of roadside scarps, edges of dirtroads, gravel-, sandor loam-pits. such places bear diverse albeit very much understudied lichen flora (see e.g. e r n s t 1993, 1995). leptogium biatorinum, sarcosagium campestre, steinia geophana and verrucaria xyloxena represent typical members of pioneer lichen communities inhabiting transient man-made habitats in eec european lowlands. all these species are probably widely distributed both regarding climatic zones and oceanic gradient. distribution of these lichens in eec europe even more strongly reflects regional activity of lichenologists, as these species are most often recorded during targeted studies of specific habitats. to date, the widest distribution in west-east direction is demonstrated by sarcosagium campestre (fig. 4), verrucaria xyloxena (fig. 5), and leptogium biatorinum (fig. 6), two latter species being recorded also outside the discussed region of eec europe, in the steppe zone in russia (ve d e n e e v 1999; m u c h n i k 2001; s h u s t o v 2002). steinia geophana is more widely distributed in a south-north direction (fig. 7): outside the discussed area its range reaches kola peninsula in russia (bilas herbarium, specimens no. 5304 and 5339). however, not all of this data is totally reliable, as at least two species can be source of taxonomic confusion. in some references (western european as well) l. biatorinum is not differentiated from l. byssinum (j ø r g e n s e n 1994). in earlier russian and ukrainian references (o k s n e r 1956; i n a s h v i l i 1975), the description of l. byssinum fig. 4. distribution of sarcosagium campestre in eastern and east central europe � modern records fig. 5. distribution of verrucaria xyloxena in eastern and east central europe � records more than 50 year old, � modern records lichens of neglected habitats 149 ta b l e 1 distribution, frequency and conservation status of the selected lichen species in eastern and east central european lowlands frequency abbreviations: vr (very rare), r (rare), rr (rather rare), rf (rather frequent), f (frequent), vf (very frequent). frequency evaluations for lithuania are given according to m o t i e j ū n a i t ė (2002a), for estonia according to r a n d l a n e and s a a g (1999), for latvia according to p i t e r ā n s ( 2 0 0 2 ) ( i n p a r t ) . in some cases parts of countries are indicated (e.g., ne north eastern, c central). conservation status abbreviations: vu (vulnerable), nt (near threatened), dd (data deficient). references for conservation information: c i e ś l i ń s k i (2003b), c i e ś l i ń s k i et al. (2003). for country abbreviations see chapter “territory under discussion”. species countries, frequency references conservation status leptogium biatorinum (nyl.) leight. p (rr), li (rr), lv (vr), r (vr) l e t t a u 1912; c e y n o w a g i e ł d o n 2001; c i e ś l i ń s k i 2003a; m o t i e j ū n a i t ė et al. 2006 sarcosagium campestre (fr.) poetsch & schied. p (rr), li (rr), lv (vr), u(vr), e(vr) l i p n i c k i 1991; f a ł t y n o w i c z 1992, 2003; k o n d r a t y u k et al. 1998; c e y n o w a g i e ł d o n 1999; r u t k o w s k i , s ł o w i k 1999; c i e ś l i ń s k i 2003a; s p a r r i u s 2003; s u i j a et al. 2005; m o t i e j ū n a i t ė et al. 2006; k u k w a , pers. com. steinia geophana (nyl.) stein p (r), li (rr), e (r), r (vr) f a ł t y n o w i c z 1993; 2003; f a d e e v a et al. 1997; c e y n o w a g i e ł d o n 1998, 1999; z a v a r z i n et al. 1999; h a l o n e n et al. 2000; k u z n e t s o v a , h i m e l b r a n t 2002; c i e ś l i ń s k i 2003a; a p t r o o t et al. 2005 p (nt in ne) verrucaria aquatilis mudd p (vr), li (rr), la (vr), r (vr) k o p a c h e v s k a y a 1977; f a ł t y n o w i c z 1992; r u t k o w s k i 1993; f a d e e v a et al. 1997; f a ł t y n o w i c z et al. 2000; k o w a l e w s k a et al. 2000; k u k w a 2000; c i e ś l i ń s k i 2003a; m o t i e j ū n a i t ė et al. 2006 p (vu, dd in ne) verrucaria hydrela ach. p (rr), li (f), la (r), e (vr), r (vr) l e t t a u 1912; p i t e r ā n s 1982; f a d e e v a et al. 1997; f a ł t y n o w i c z et al. 2000; h a l o n e n et al. 2000; k o w a l e w s k a et al. 2000; k u k w a 2000; f a ł t y n o w i c z , k r ó l a k 2001; c i e ś l i ń s k i 2003a; j a n d o , k u k w a 2003; m o t i e j ū n a i t ė et al. 2006 p (vu, not red listed in ne) verrucaria praetermissa (trevis.) anzi p (vr), li (rf), la (vr), e (vr), r (vr) h a l o n e n et al. 2000; a n d r e e v 2002; c i e ś l i ń s k i 2003a; m o t i e j ū n a i t ė et al. 2006 p (nt, dd in ne) verrucaria xyloxena norman p (rf, common in c), li (rr), e (vr), r (vr) o r a n g e 1991; c e y n o w a g i e ł d o n 1998; r a n d l a n e , s a a g 1999; c i e ś l i ń s k i 2003a; k u z n e t s o v a , pers. com. 150 j. motiejūnaitė can accomodate both species. in estonia only l. byssinum is recorded (r a n d l a n e , s a a g 1999) basing only on literature data, thus the identity of the species is questionable. illustrations and descriptions in polish references (n o w a k , to b o l e w s k i 1975; c e y n o w a g i e ł d o n 1999) define both lichens very well. l e t t a u (1912) also circumscribed these two species. part of the reports of v. xyloxena southeastwards of the discussed area (voronezh, volgograd, samara, saratov, uljanovsk regions of russia) might be doubtful – the species is reported as saxicolous (m u c h n i k 2001; s h u s t o v 2002). frequencies of transient habitat species in eastern europe reflect general recording – locally they are reported as quite common, but data is totally absent from most of the territory. in most countries they are reported to be rare or very rare (ta b . 1). regardless record scarcity, it can be assumed that they are much more frequent, as traditionally there are numerous habitats suitable for these lichens, such as sand-, graveland loam-pits which are present in almost every village, dirtroads, road scarps, etc. and recording does not show any tendencies of decrease. even a cursory search through such habitats can result in a variety of finds (e.g. m o t i e j ū n a i t ė 2002b; s p a r r i u s 2003), and intensive targeted search can totally change frequency patterns (c e y n o w a g i e ł d o n 1998, 1999, 2001). there are different opinions concerning the dynamics of these species in europe. in j ø r g e n s e n (1994) and j ø r g e n s e n and m o t i e j ū n a i t ė (2005) species of transitional habitats are presumed to be decreasing due to modern agricultural methods. wo o d s and c o p p i n s (2003) express opinion that species of transient habitats should be excluded from the red-lists, though they evaluate verrucaria xyloxena as critically endangered in the same reference. however, in some countries lichens of transient habitats are given threat status: leptogium biatorinum is red listed in denmark, finland (v i t i k a i n e n et al. 1997) and in lowland lands of germany (categories range from extinct to poorly known); sarcosagium campestre and steinia geophana – in denmark and in lowland lands of germany (variable categories); verrucaria xyloxena is red-listed in germany as data deficient (not recorded in lowland lands). in eec europe, lichens of transient habitats can hardly be qualified as threatened though steinia geophana is red listed in ne poland. on the contrary, part of these lichens might belong to spreading species, as they occupy niches that are naturally scarce in the discussed region and appear mainly in connection with human activities. fig. 6. distribution of leptogium biatorinum in eastern and east central europe � records more than 50 year old, � modern records fig. 7. distribution of steinia geophana in eastern and east central europe � records more than 50 year old, � modern records lichens of neglected habitats 151 conclusions as the examples in this paper have shown, there are large spatial gaps in recording lichens of aquatic and transitional habitats in eec europe, making these lichens nationally rare in almost all countries in the discussed region. also there exists certain degree of taxonomic confusion pertaining these species. lichen floras, resulting from species inventories make base for environmental evaluation and monitoring programmes, land manegement policy making, etc. disregard of certain habitats or species, or on the contrary, detailed study of all habitat types and all species present might lead to very different results of lichen inventories (see e.g. l õ h m u s et al. 2003) and subsequently, to erroneous interpretations of monitoring evaluation results. in wider scale, both historical and present understudy of some habitats leads to the dearth of knowledge on true lichen distribution areas and tendencies of their decrease or spreading and often disable biodiversity monitoring projects in larger geographical scale. acknowledgements: i would like to express my sincere thanks to my colleagues prof. k. czyżewska (łódż), dr. m. kukwa (gdańsk), dr. v. golubkov (grodno), mr. d. himelbrant and ms. e. kuznetsova (st. peterburg) for the data they supplied and allowed to use in the paper and their valuable comments on the manuscript. references a n d r e e v m . p. 2002. dopolnenije k likhenoflore kaliningradskoi oblasti rossii: lishainiki kamen istykh substratov. novosti sist. nizsh. rast. 36: 68 72. a p t r o o t a . , c z a r n o t a p. , j ü r i a d o i . , k o c o u r k o v á j . , k u k w a m . , l õ h m u s p. , p a l i c e z . , r a n d l a n e t. , s a a g l . , s é r u s i a u x e . , s i p m a n h . , s p a r r i u 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(in:) n . b . b a l a s h o v a , a . a . z a v a r z i n (eds.), biodiversity of the leningrad region (algae. fungi. lichens. bryophytes. invertebrates. fishes and pisciformes): 205 260. st. petersburg university press, st. petersburg. 154 j. motiejūnaitė porosty słabo poznanych siedlisk na niżu europy środkowo-wschodniej i wschodniej s t r e s z c z e n i e w pracy przedstawiono rozmieszczenie, frekwencję i stopień zbadania porostów sie dlisk wodnych i krótkotrwałych w obrębie strefy leśnej niżu europy środkowo wschodniej i wschodniej: polski, litwy, łotwy, estonii i białorusi oraz ukrainy i europejskiej części rosji. oba typy siedlisk są bardzo słabo rozpoznane w tej części europy. zagadnienie to omówio no na przykładzie wybranych gatunków efemerycznych: leptogium biatorinum, sarcosagium campestre, steinia geophana i verrucaria xyloxena oraz wodnych: verrucaria aquatilis, v. hydrela i v. praetermissa. w skali wielkoprzestrzennej widoczne są luki w rozmieszczeniu badanych gatunków (figs 1 7). ustalenie ich rzeczywistego rozmieszczenia i dynamiki rozprzestrzenienia jest, jak na razie, trudne. celowe poszukiwanie odpowiednich siedlisk i charakterystycznych im gatunków sygnalizuje, że mogą one być nawet częste w regionie. 2014-01-01t11:44:00+0100 polish botanical society the therapeutic potential of truffle fungi: a patent survey 305this is an open access article distributed under the terms of the creative commons attribution 3.0 license (creativecommons.org/licenses/by/3.0/), which permits redistribution, commercial and non-commercial, provided that the article is properly cited. © the author(s) 2014 published by polish botanical society acta mycologica review acta mycol 49(2):305–318 doi: 10.5586/am.2014.022 received: 2014-05-21 accepted: 2014-11-24 published electronically: 2014-12-31 the therapeutic potential of truffle fungi: a patent survey małgorzata gajos1*, florian ryszka2, joerg geistlinger3 1 department of biomedical computer systems, university of silesia, będzińska 39, 41-200 sosnowiec, poland 2 pharmaceutical research and production plant “biochefa”, kasztanowa 3, 41-205 sosnowiec, poland 3 department of agriculture, ecotrophology and landscape development, anhalt university of applied sciences, strenzfelder allee 28, 06406 bernburg, germany abstract the purpose of this article is to research and retrieve patent information regarding the therapeutic use of truffles. truffles have a unique value as a foodstuff and impact positively on human health and well-being. they are applied in such industries as the pharmaceutical industry and the cosmetic industry. patent documentation available in the espacenet network and the patentscope service were analyzed by key word and patent specifications were examined to describe state of the art and to identify scientific research trends in therapeutic applications of truffles. medicinal properties of truffles such as the anticancer or cardiovascular effect, a reduction in blood lipids, immunological resistance and increased energy were identified. other therapeutic benefits include sedative action, prevention of hormonal imbalances in women, pre-menopause symptom relief, senile urethritis and prostate disorders, sleep disorders and increased absorption of calcium from milk. truffles can also be used to alleviate symptoms of milk intolerance such as diarrhoea or bloating, to ease rheumatic pains and to treat and prevent further development or recurrence of senile cataract. keywords: truffle fungi; tuber; therapeutic potential of truffles; patent survey introduction the number of patents granted annually to inventions is an important indicator of technological development [1]. in order to obtain patent protection, each invention should * corresponding author. email: malgorzata.gajos@us.edu.pl handling editor: maria rudawska http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.2014.022 mailto:malgorzata.gajos%40us.edu.pl?subject=am.2014.022 306© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles be characterized by patentability, determined by novelty, inventive step and industrial application. this article reviews patent research into truffle (tuber)-related inventions. it addresses the important role of truffles in the food industry and documents their positive effects on human health and well-being as well as their usefulness in the pharmaceutical industry. however, truffles are some of the most expensive organic products in the world due to limited resources and difficulties in obtaining them. patent research is a set of activities consisting of searching and analyzing patent information. it comprises searches of patent literature and documentation that are further analyzed for relevant information. conclusions based on patent research can be successfully used to evaluate practicability of applying for patent protection for a specific solution, to provide solutions of international technical standard permitting patent protection, to select scientific research directions, to secure presumption that a technical solution does not infringe on patent rights of third persons and to use known solutions when implementing research objectives within legally permissible imitation. a recent publication by gajos and hilszczańska [2] surveys the state of the art and research interests regarding truffles. the purpose of the present article was to analyze truffle patent information. patent documentation was researched through patent databases and patent specifications. patented and patent-applied developments as well as specific thematic scopes of implemented invention proposals were explored, with the scientific and technical information being studied. such patent research primarily evaluates the state of the art in truffle research. there are more than 100 truffle species belonging to the tuberales worldwide. the most economically significant truffles belong to the genus tuber and grow, e.g., in southern europe, particularly in northern spain, italy and greece, in southern france and croatia. truffles contain a variety of biochemical components, including steroids, pheromones, flavonoide, anthocyanine, carotenoide, oligosaccharide and volatile organic compounds (vocs) such as dimethyl-sulfide and -bisulfide, butanedione, butyrate, hexenone, ethyl-, methyland propylphenole and methylthiomethane. to date they are widely unexplored and unexploited resources that have a therapeutic potential as they contain antioxidant, antimicrobial, antiviral, antimutagenic, anticarcinogenic, immune-modulating, hepatoprotective, antidepressant and sedative bioactive substances [3]. truffles have protective effects due to their antioxidant [4] and antiradical properties [5]. they contain a variety of phenolic substances, which are very efficient scavengers of peroxyl radicals [4]. a previous report showed the expression of some 120 antioxidant enzymes [6]. correspondingly, truffles show strong inhibition of lipid oxidation, which indicates effective antioxidant activity [7]. the antioxidant-rich aqueous extract of desert truffle terfezia claveryi was demonstrated to have a very powerful hepatoprotective activity when evaluated in rats using the potent hepatotoxin carbon tetrachloride [8]. a few studies discuss the antimicrobial importance of truffles [9, 10]. activity against gram(−) and gram(+) bacteria has been reported [11]. truffle extracts showed in-vitro activity against chlamydia trachomatis, staphylococcus aureus and pseudomonas aeruginosa. antiviral activity was reported by hussan and al-ruqaie [12]. antimutagenicity was studied with tuber aestivum (summer truffle) extracts and irradiation of bacteria with the application of the mutagens nitroflurene and sodium azide [13]. anti-inflammatory effects were determined as inhibitory effects on cyclooxygenase-2 307© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles (cox2), an enzyme that plays an important role in the inflammatory process [14]. intracellular polysaccharides isolated from the fruiting body of tuber sinense have been shown to exhibit immuno-modulation and antitumour activities [15]. the influence on tumour cell growth was detected by ferreira et al. [16] and ng and wong [17]. possible bioactive substances were classified as quinines, isoflavones, steroids, ceramides and catechols. a new polyhydroxy sterol glycoside named tuberoside has been isolated from fruiting bodies of tuber indicum. these compounds are assumed to be structural constituents of cellular membranes and precursors of steroid hormones that antagonise anxiety and act as sedatives by positively modulating gaba receptors [18]. a genetic analysis of tuber progressed substantially with the publication of the total genome sequence of tuber melanosporum (black perigord truffle) [19]. it is by far the largest genome sequence (125 mb) ever obtained from an ascomycete fungus but, in contrast, only 7500 protein-coding genes were found, with large gene families almost completely missing. moreover, simple sequence repeats are highly abundant in this genome [20], which offers an excellent opportunity for the development of molecular markers that can be applied in further genetic analyses of various species or genotypes. information on the transcriptome exists from tuber borchii (bianchetto truffle) regarding differentially expressed genes in vegetative mycelium and fruiting bodies [21]. genomic information regarding the genus tuber is clearly quite abundant and genes for biochemical pathways involved in the synthesis of bioactive molecules (anti-inflammatory, antioxidant, etc.) and vocs that are responsible for the different tastes, odors, scents and aromas of tuber could be identified based on it. truffles can be commercialized as food additives or flavors as well as fragrance in cosmetics industries [22]. furthermore, it has been shown that isolated tuber genes can be expressed in heterologous expression systems like escherichia coli and yeast [23,24]. this is a prerequisite for the biotechnological production of bioactive compounds independent of natural truffle grounds. due to a decline in natural tuber populations, caused by intensive collection of fruiting bodies for cooking, attempts have been made to cultivate tuber species as fermentation mycelia. it has been shown that mycelia grown in-vitro had elevated sterol content as compared to fruiting bodies collected from the wild [25]. material and methods espacenet and patentscope as well as patent specifications for submitted and patented truffle-related developments were used in this study. in october 1998, the european patent office (epo) launched esp@cenet, a network of free patent databases containing patent documents (patents and patent applications) from europe, japan, the u.s. and the world intellectual property organization (wipo) [26]. patentscope (wipo) is a free search service offered by the world intellectual property organization. it includes many national patent collections, including some of rare coverage. patent specifications are basic sources of scientific-technical information. they are carriers of technical, legal, economic, sociological and organizational information as well as personal information regarding inventors. a patent specification is a document that consists of a bibliographical part, a description of the invention, patent claims and illustrations [27]. 308© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles codes found in patent descriptions in brackets or circles are designed to facilitate their drafting, information searches and patent research performance. international unification of the bibliographical description is independent of the language in which the bibliographical description is formulated as well as of the composition and sequence of factors, which determine a country-specific patent system. associated standards st. recommendations and guidelines as elaborated by the wipo are published in the “handbook on industrial property information and documentation” (e.g. st.3 two-letter codes for the representation of states, other entities and organizations, e.g. cn – china, ep – european patent office, wo world intellectual property organization; st.9 bibliographical data on and relating to patents; st.16 identification of different kinds of patent documents, e.g., a first publication level patent application, dependence on national system could also be e.g. a1), b – second publication level patent specification, dependence on national system could also be e.g. b1) [28]. patenting solutions for truffles can be recognized based on analyses of inventions, patent claims and drawings. the classification code in the bibliographical part, which is also known as the international patent classification (ipc), consists of the symbol of a section (a–h). each section is subdivided into classes (each class symbol consists of the section symbol followed by a two-digit number). each class comprises one or more subclasses (each subclass symbol consists of the class symbol followed by a capital letter). each class and subclass has a title. each subclass is broken down into subdivisions referred to as “groups” which are either main groups or subgroups [29]. bibliographical data studies of patent specifications can help set research directions, e.g. to classify objects according to the ipc, and establish countries, companies and investors leading in the field of truffles or names of internationally renowned experts who can be patent creators. patent research is mainly conducted by patent offices (in connection with the consideration of patent applications) as well as patent attorneys and invention services (in connection with technical innovations being applied for patenting and implementation of new technology and service products). in order to recognize the state of the art and to delineate scientific research directions, patent research can also be performed by scientific practitioners, especially when there is access to internet patent databases. the advantage of electronic databases is that they offer easy access to thematic information by keyword search. patent research includes: (i) basic research comprising patent discernment (stateof-the-art research), presumption that a given technical solution does not infringe on patent rights of third persons, patentability research and comprehensive patent research; (ii) special research for technology development planning and forecasting, standardisation papers, foreign trade and innovations development purposes. the article presents research into the state-of-the-art in relation to truffles. such studies should precede any scientific research of technical nature. the objective of patent research is to present developments in a specific domain of technology and technical advances. the time scope is based on the development rate of a specific discipline: five to six years in rapidly growing domains and slightly more for domains which grow more slowly. since this article is meant to recognize the broad status of truffle-related developments, no time constraints were introduced and espacenet and patentscope bases were searched in their entirety. 309© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles the following patent research methods are distinguished: object (by classification or keyword), subject (by company name or inventor’s personal name), patents obtained for the same invention in different countries (by priority dates), mixed (using elements of the above methods). this article uses the object method; research object is classified according to the ipc and keyword search was used for a research topic. an analysis was run for identified patent specifications and the research status was determined. next the subject method was applied. the country of origin was established for leading institutions in the research domain covered by this article. finally, possible research directions were set for truffle research. results and discussion the international patent classification represents the whole body of knowledge, which may be regarded as proper to the field of patents for invention. the following sections and classes were investigated in this article: (i) a (human necessities) – a01 (agriculture, forestry, animal husbandry, hunting, trapping, fishing); a23 (foods or foodstuffs, their treatment); a61 (medical or veterinary science, hygiene); (ii) c (chemistry; metallurgy) – c07 (organic chemistry); c11 (animal or vegetable oils, fats, fatty substances or waxes, fatty acids thereof, detergents, candles); c12 (biochemistry, beer, spiritus, wine, vinegar, microbiology, enzymology, mutation or genetic engineering); (iii) g (physics) – g01 (measuring, testing). as of december 2012, a total of 29 documents regarding truffle inventions (patent applications and patent specifications or only patent applications) concerning medicine, health, pharmaceuticals and diet supplements were found. patent application (a, a1) follows formal and legal research, patent specification (b) follows formal, legal and substantive research (patentability). results are presented in a table (tab. 1). patent information regarding all twenty nine inventions belongs to the state of the art but only six inventions were granted a patent (with five patents belonging to chinese investors and one belonging to an italian investor); the procedure has not been closed for one patent yet and the remaining inventions were not granted a patent. titles of inventions and sources of patent applications or patent specifications are given in a table (tab. 2). anticancer properties (invention 5) one of the many benefits of truffles used in the pharmaceutical industry and medicine is their anticancer action as reflected in the inhibition of tumor cell growth. medical properties of wild truffles are stronger as compared to those found in other species of fungi such as in ganoderma lucidum (lingzhi mushroom) and cordyceps militaris (scarlet caterpillar club mushroom). the process of preparing a natural product from wild truffles, black truffle (t. melanosporum) and brown truffle, comprises adding the aforementioned fungal species to distilled water, inactivation at 121–125° c under a pressure of 1 atm. for 2 to 3 hours followed by addition of alcohol and uniform mixing. the solution is kept 310© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles in ve nt io n nu m be r c ou nt ry c od e fo r pu bl is hi ng , nu m be r an d ty pe c od e of do cu m en t (p at en t sp ec ifi ca ti on ) d at e th e pa te nt is is su ed c ou nt ry c od e fo r pu bl is hi ng , nu m be r an d ty pe c od e of do cu m en t (p at en t ap pl ic at io n) p ub lic at io n da te of th e in ve nt io n do cu m en ta ti on (a pp lic at io n) p ri or it y da te in te rn at io na l pa te nt c la ss ifi ca ti on ip c v er si on 8 in ve nt or medicine and health – pharmaceuticals, diet supplements (29) 1 cn102224872 (a) 2011-10-26 2011-04-28 a23f5/24 a23f5/38 a23f5/40 cn 2 cn102145019 (b) 2012-10-10 cn102145019 (a) 2011-08-10 2011-03-29 a23c9/152 a23l1/29 a23l2/38(+31) cn 3 cn101965972 (a) 2011-02-09 2010-10-12 a23l1/20 a23l1/28 a23l1/29 cn 4 cn101911989 (a) 2010-12-15 2010-08-03 a23f3/14 cn 5 cn101732362 (b) 2011-06-15 cn101732362 (a) 2010-06-16 2008-11-21 a61k36/06 a61p35/00 a61p37/04 cn 6 cn101313920 (a) 2008-12-03 2008-06-25 a23l1/30 a61k36/06 a61p13/02(+3) cn 7 cn101268831 (a) 2008-09-24 2008-04-08 a23l1/28 a23l1/29 a61k36/06(+3) cn 8 it1190435 (b) 1988-02-16 1985-12-13 a23l it 9 cn102551066 (a) 2012-07-11 2012-03-26 a23l1/28 a23l1/29 cn 10 cn102334549 (a) 2012-02-01 2011-06-08 a23c9/12 cn 11 cn102250709 (b) 2012-12-12 cn102250709 (a) 2011-11-23 2011-06-08 c12g1/022 c12r1/645 cn 12 cn102224853 (a) 2011-10-26 2011-06-08 a23d9/007 a23d9/04 a23l1/226 cn 13 cn102090455 (a) 2011-06-15 2010-09-25 a23c9/152 a23c9/18 cn 14 cn102771860 (a) 2012-11-14 2012-07-20 a01g1/04 a23l1/28 a23l1/29 a23l2/38 cn 15 cn102771773 (a) 2012-11-14 2012-07-20 a01g1/04 a23l1/28 a23l1/30 cn 16 cn102771857 (a) 2012-11-14 2012-07-20 a23l1/28 a23l2/38 a23l2/52 cn 17 cn102771855 (a) 2012-11-14 2012-07-20 a01g1/04 a23l1/29 a23l2/38 cn 18 cn102771762 (a) 2012-11-14 2012-07-20 a01g1/04 a23l1/28 a23l1/29 cn 19 cn102640820 (a) 2012-08-22 2012-04-25 a23f3/14 cn 20 cn102599514 (a) 2012-07-25 2012-03-26 a23l1/29 cn 21 cn102224873 (b) 2012-12-05 cn102224873 (a) 2011-10-26 2011-05-06 a23f5/40 a23f5/42 cn 22 cn102224925 (b) 2012-12-12 cn102224925 (a) 2011-10-26 2011-05-06 a23l1/29 c12n1/14 cn 23 cn102199547 (a) 2011-09-28 2011-04-28 c12n1/14 c12r1/645 cn tab. 1 patent research about truffles in espacenet and patentscope database. 311© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles overnight at 4 to 10°c, then subjected to centrifugation 1000 to 1200 rpm, 10–15 min, and the resulting supernatant is lyophilized. cardiovascular effect (invention 4, 19) another example of the operation of active substances in fungi is their effect on the cardiovascular system. tea extracts composed of truffles, lingzhi mushrooms and black beans can be applied to prevent and treat cardiovascular and cerebrovascular diseases. the preparation method includes: weighing 15–20 parts of sun-dried tea, 2–6 part truffles, 2–5 part lingzhi mushrooms and 2–5 parts of black beans, followed by ethanol extraction of fungi and black beans, washing the extract with water, combining the washed extracts with tea and fermentation (invention 4). pu-erh tea compositions containing arctium lappa (greater burdock 40–60%) and truffles (20–40%) are used to lower blood pressure. these teas are brewed for 3 minutes in 250 ml of water at 93°c. one bag may be brewed from 3 to 5 times (invention 19). reduction in blood fats (invention 12, 19, 24) pu-erh tea compositions containing greater burdock and truffles also demonstrate an effect of lowering blood lipid levels (invention 19). a similar behavior is demonstrated by edible oil containing vegetable oil or animal fats and truffle-derived oil. the volume ratio of the above-mentioned fats/oils in the raw material is 1:0.1–1 (invention 12). a preparation containing red tea and truffles lowers the level of blood fats (invention 24) and can also be used as edible oil the composition of which is based on vegetable or animal fats and fat derived from truffles. the volume ratio of the above-mentioned fats in the raw material is 1:0.1–1 (invention 12). a preparation containing red tea and truffles has also been proven to have this effect. raw material of this preparation contains 1–20 parts (by in ve nt io n nu m be r c ou nt ry c od e fo r pu bl is hi ng , nu m be r an d ty pe c od e of do cu m en t (p at en t sp ec ifi ca ti on ) d at e th e pa te nt is is su ed c ou nt ry c od e fo r pu bl is hi ng , nu m be r an d ty pe c od e of do cu m en t (p at en t ap pl ic at io n) p ub lic at io n da te of th e in ve nt io n do cu m en ta ti on (a pp lic at io n) p ri or it y da te in te rn at io na l pa te nt c la ss ifi ca ti on ip c v er si on 8 in ve nt or 24 cn102058728 (a) 2011-05-18 2010-11-16 a23f3/14 a23l1/29 a61k35/64(+9) cn 25 cn101856370 (a) 2010-10-13 2010-07-07 a23l1/29 a61k36/068 a61k36/28(+7) cn 26 cn101892143 (a) 2010-11-24 2009-05-20 c12g3/04 cn 27 jp2002249438 (a) 2002-09-06 2001-02-22 a23l1/30 a61q1/2 jp 28 jp2002173441 (a) 2002-06-21 2000-09-27 a23l1/30 a61q 19/10 jp 29 us20120039927 a1 wo2012021466 2012-02-16 a61p27/12 a61k 36/062 us tab. 1 (continued) source: the author’s eleboration based on espacenet and patentscope databases. 312© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles inv. no. title of invention source of patent information regarding invention a/b medicine and health – pharmaceuticals, diet supplements (29) 1 health food with truffles and coffee beans as raw materials http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0111026&cc=cn&nr=102224872a&kc=a a 2 medicinal composition containing truffles, vitamins and mineral substances http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0110810&cc=cn&nr=102145019a&kc=a b 3 health-care food made of truffles and beans http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0110209&cc=cn&nr=101965972a&kc=a a 4 truffle health-care tea and preparation method thereof http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0101215&cc=cn&nr=101911989a&kc=a a 5 application of wild truffle in preparing medicine with anticancer and immunological functions http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0100616&cc=cn&nr=101732362a&kc=a b 6 application of truffle extract in preparing foods, medicine for treating endocrine dyscrasia diseases http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0081203&cc=cn&nr=101313920a&kc=a a 7 truffle hydrophilic solvent extract and application of the same in preparing food, health food, and medicament for treating prostate gland disease etc. http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0080924&cc=cn&nr=101268831a&kc=a a 8 solid truffle product http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=1 9880216&cc=it&nr=1190435b&kc=b b 9 health-care food containing maca, cordyceps and truffle http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0120711&cc=cn&nr=102551066a&kc=a a 10 dairy product with raw materials containing truffle http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0120201&cc=cn&nr=102334549a&kc=a a 11 beverage prepared from raw material containing truffle http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0111123&cc=cn&nr=102250709a&kc=a b 12 edible oil with raw material containing truffle oil http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0111026&cc=cn&nr=102224853a&kc=a a 13 truffle-containing milk http://worldwide.espacenet.com/publicationdetails/ biblio?adjacent=true&locale=en_ep&ft=d&date=2 0110615&cc=cn&nr=102090455a&kc=a a 14 truffle health drink prepared by liquid-submerged fermentation and preparation method of truffle health drink http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=5&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20121 114&cc=cn&nr=102771860a&kc=a a tab. 2 titles of truffle-related inventions based on espacenet and patentscope databases. http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20111026&cc=cn&nr=102224872a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20111026&cc=cn&nr=102224872a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20111026&cc=cn&nr=102224872a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20110810&cc=cn&nr=102145019a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20110810&cc=cn&nr=102145019a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20110810&cc=cn&nr=102145019a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20110209&cc=cn&nr=101965972a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20110209&cc=cn&nr=101965972a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20110209&cc=cn&nr=101965972a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20101215&cc=cn&nr=101911989a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20101215&cc=cn&nr=101911989a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20101215&cc=cn&nr=101911989a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20100616&cc=cn&nr=101732362a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20100616&cc=cn&nr=101732362a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20100616&cc=cn&nr=101732362a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20081203&cc=cn&nr=101313920a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20081203&cc=cn&nr=101313920a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20081203&cc=cn&nr=101313920a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20080924&cc=cn&nr=101268831a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20080924&cc=cn&nr=101268831a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20080924&cc=cn&nr=101268831a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=19880216&cc=it&nr=1190435b&kc=b http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=19880216&cc=it&nr=1190435b&kc=b http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=19880216&cc=it&nr=1190435b&kc=b http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20120711&cc=cn&nr=102551066a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20120711&cc=cn&nr=102551066a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20120711&cc=cn&nr=102551066a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20120201&cc=cn&nr=102334549a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20120201&cc=cn&nr=102334549a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20120201&cc=cn&nr=102334549a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20111123&cc=cn&nr=102250709a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20111123&cc=cn&nr=102250709a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20111123&cc=cn&nr=102250709a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20111026&cc=cn&nr=102224872a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20111026&cc=cn&nr=102224872a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20111026&cc=cn&nr=102224872a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20110615&cc=cn&nr=102090455a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20110615&cc=cn&nr=102090455a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?adjacent=true&locale=en_ep&ft=d&date=20110615&cc=cn&nr=102090455a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=5&nd=3&adjacent=true&locale=en_ep&ft=d&date=20121114&cc=cn&nr=102771860a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=5&nd=3&adjacent=true&locale=en_ep&ft=d&date=20121114&cc=cn&nr=102771860a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=5&nd=3&adjacent=true&locale=en_ep&ft=d&date=20121114&cc=cn&nr=102771860a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=5&nd=3&adjacent=true&locale=en_ep&ft=d&date=20121114&cc=cn&nr=102771860a&kc=a 313© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles inv. no. title of invention source of patent information regarding invention a/b 15 truffle health-care product containing traditional chinese medicine extractive and preparation method thereof http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=6&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20121 114&cc=cn&nr=102771773a&kc=a a 16 truffle health-care drink prepared through liquid submerged fermentation and preparation method of truffle health-care drink prepared through liquid submerged fermentation http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=7&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20121 114&cc=cn&nr=102771857a&kc=a a 17 truffle health care product and preparation method thereof http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=8&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20121 114&cc=cn&nr=102771855a&kc=a a 18 truffle health-care product prepared through liquid submerged fermentation and preparation method of truffle health-care product prepared through liquid submerged fermentation http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=9&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20121 114&cc=cn&nr=102771762a&kc=a a 19 formula of health tea with truffle and burdock and production method of health tea with truffle and burdock http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=14&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20120 822&cc=cn&nr=102640820a&kc=a a 20 health food containing maca and truffle http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=15&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20120 725&cc=cn&nr=102599514a&kc=a a 21 method for preparing food from raw materials comprising coffee beans http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=21&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20111 026&cc=cn&nr=102224873a&kc=a b 22 method for preparing food from truffle and coffee beans as raw materials http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=22&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20111 026&cc=cn&nr=102224925a&kc=a b 23 truffle fermentation culture medium and preparation method of truffle http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=23&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20110 928&cc=cn&nr=102199547a&kc=a a 24 medicament with raw materials containing red tea and truffle http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=33&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20110 518&cc=cn&nr=102058728a&kc=a a 25 medicine combination capable of regulating immunity http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=38&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20101 013&cc=cn&nr=101856370a&kc=a a tab. 2 (continued) http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=6&nd=3&adjacent=true&locale=en_ep&ft=d&date=20121114&cc=cn&nr=102771773a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=6&nd=3&adjacent=true&locale=en_ep&ft=d&date=20121114&cc=cn&nr=102771773a&kc=a 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http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=33&nd=3&adjacent=true&locale=en_ep&ft=d&date=20110518&cc=cn&nr=102058728a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=33&nd=3&adjacent=true&locale=en_ep&ft=d&date=20110518&cc=cn&nr=102058728a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=38&nd=3&adjacent=true&locale=en_ep&ft=d&date=20101013&cc=cn&nr=101856370a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=38&nd=3&adjacent=true&locale=en_ep&ft=d&date=20101013&cc=cn&nr=101856370a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=38&nd=3&adjacent=true&locale=en_ep&ft=d&date=20101013&cc=cn&nr=101856370a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=38&nd=3&adjacent=true&locale=en_ep&ft=d&date=20101013&cc=cn&nr=101856370a&kc=a 314© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles weight) of red tea, 5–50 parts (by weight) of truffles and 1–30 parts (by weight) of one or more of the following: mistletoe, medlar, chrysanthemum, honeysuckle flower, honey, ginseng, the stem and root of dwarf big blue lilyturf (invention 24). immunological resistance (invention 1–5, 7, 10–11, 19, 21–22, 24–26) pharmaceutical research has shown that a diet, which includes truffles, helps to increase resistance and fights fatigue. foodstuffs most commonly rely on a combination of truffles and coffee beans (invention 1, 3, 21–22) or [common] beans (invention 3). in both cases, the weight ratio of the abovementioned components in the raw material should be 5–50 parts truffles and 10–100 of coffee beans or [common] beans (invention 1, 3). dairy products and beverages containing grapefruit juice provide further examples of applications of substances present in fungi. milk products are based on milk and truffle fermentation wort or broth where both the volume and weight ratios of the raw material are (1–10) : (1–20). in beverages, the grapefruit juice volume ratio to the truffle fermentation broth in the raw material is (1–10) : (1–20) while the weight ratio to the truffle extract equals (1–10) : (1–20). it has been proven that both of these products have an anti-aging effect and increase somatic resistance (invention 10–11). the aforementioned tea varieties containing truffle, lingzhi mushroom and black bean extracts or greater burdock and truffles also have a positive impact on immunological resistance (invention 4, 19). an effect on immunological resistance growth is also exhibited by pharmaceutical preparations, which contain a truffle extract alone, or in combination with other substances. exemplary preparations can consist of truffles, vitamins and minerals in a weight ratio equal to 0.1–10 parts, 0.005–3 parts and 0.05–3 parts, respectively, as well inv. no. title of invention source of patent information regarding invention a/b 26 method for preparing medicinal liquor http://worldwide.espacenet.com/publicationdetails/ biblio?db=worldwide.espacenet.com&ii=43&nd=3 &adjacent=true&locale=en_ep&ft=d&date=20101 124&cc=cn&nr=101892143a&kc=a a 27 age retardant for living organism http://patentscope.wipo.int/search/en/detail.jsf ?doci d=jp67349238&recnum=34&office=&querystring= alltxt%3a%28truffle%29&prevfilter=&sortoptio n=relevance&maxrec=385 a 28 agent for suppressing or preventing oxidation of organism http://patentscope.wipo.int/search/en/detail.jsf ?doci d=jp67273260&recnum=37&office=&querystring= alltxt%3a%28truffle%29&prevfilter=&sortoptio n=relevance&maxrec=385 a 29 use of extracts of pezizaceae in the prevention and/or treatment of senile cataracts http://patentscope.wipo.int/search/en/detail.jsf ?doci d=us73487155&recnum=105&office=&querystring =alltxt%3a%28truffle%29&prevfilter=&sortopt ion=relevance&maxrec=385 a1 tab. 2 (continued) source: the author’s eleboration based on espacenet and patentscope databases. inv. no. – invention number; a – application; b – patent. http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=43&nd=3&adjacent=true&locale=en_ep&ft=d&date=20101124&cc=cn&nr=101892143a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=43&nd=3&adjacent=true&locale=en_ep&ft=d&date=20101124&cc=cn&nr=101892143a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=43&nd=3&adjacent=true&locale=en_ep&ft=d&date=20101124&cc=cn&nr=101892143a&kc=a http://worldwide.espacenet.com/publicationdetails/biblio?db=worldwide.espacenet.com&ii=43&nd=3&adjacent=true&locale=en_ep&ft=d&date=20101124&cc=cn&nr=101892143a&kc=a http://patentscope.wipo.int/search/en/detail.jsf?docid=jp67349238&recnum=34&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=jp67349238&recnum=34&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=jp67349238&recnum=34&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=jp67349238&recnum=34&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=jp67273260&recnum=37&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=jp67273260&recnum=37&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=jp67273260&recnum=37&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=jp67273260&recnum=37&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=us73487155&recnum=105&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=us73487155&recnum=105&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=us73487155&recnum=105&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 http://patentscope.wipo.int/search/en/detail.jsf?docid=us73487155&recnum=105&office=&querystring=alltxt%3a%28truffle%29&prevfilter=&sortoption=relevance&maxrec=385 315© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles as combinations of truffles and cordyceps sinensis (caterpillar mushrooms) in a weight ratio equal to 0.1–5 parts and 1–10 parts, respectively (invention 2, 25). vigour growth (invention 1, 9, 20, 25) it has been shown that dietary supplements containing lepidium meyenii (maca), truffles and, optionally, scarlet caterpillar club mushroom influence the growth and strength of the body. the weight percentage of these ingredients should be 15–70%, 5–35% and 5–35%, respectively. the supplement is prepared as follows: alcohol extraction of the active ingredient of dry or fresh maca root powder and preparation of truffle and scarlet caterpillar club fungus powder in accordance with generally accepted procedures. purified powder derived from the fruit of phyllanthus emblica (indian gooseberry) or starch is auxiliary materials used in the making of the preparation (invention 9, 20). anti-aging benefits (invention 10, 11, 27) pharmacological studies have found that both milk products and beverages containing grapefruit juice in combination with truffles have anti-aging properties in addition to the impact on immunological resistance. in these studies, dairy products are based on milk and truffle fermentation wort while beverages are based on grapefruit juice and truffle fermentation wort or truffle extract (invention 10–11). other in addition, fungi-derived substances manifest a range of applications not mentioned above. these include but are not limited to: (i) sedative action (truffles, lingzhi mushroom; invention 4); (ii) prevention and treatment of hormonal imbalances in women, premenopausal symptoms, senile urethritis and sleep disorders (truffles; invention 6); (iii) treatment of male sexual dysfunctions and prostate disorders (truffles; invention 7); (iv) alleviation of intolerance symptoms to milk such as diarrhoea or bloating and increased absorption of calcium from milk (truffles; invention 13), (v) easing of rheumatic pains (truffles; invention 19), (vi) treatment and prevention of further development or recurrence of senile cataract (truffles; invention 29). conclusions the largest group of patents involves immunological resistance. it has been demonstrated that a diet containing truffles increases resistance and fights fatigue. there are a variety of applied combinations of truffles with coffee (invention 1, 3, 21–22), beans (invention 3), dairy products and beverages with a grapefruit-juice base (invention 10–11). a mixture of truffles and grapefruit juice also has anti-aging properties. immunological resistance is strengthened by compositions of tea varieties, which contain extracts of truffles, lingzhi mushroom and black beans or greater burdock and truffles (invention 4, 19). truffle extract is also effective when combined with vitamins or minerals; a combination of truffles and caterpillar mushrooms has beneficial applications (invention 2, 26). 316© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles the next group includes patents relating to increased vigour and vitality. these are dietary supplements containing maca, truffles and scarlet caterpillar club mushrooms, which are thought to affect bodily growth and strength. the formulation is also added to powdered fruit of nepalese/indian gooseberry or starch (invention 9, 20). anticancer properties are shown by aqueous extracts of black truffle (t. melanosporum) and brown truffle (invention 5). extracts containing truffles, lingzhi mushrooms and black beans are used in the treatment of cardiovascular and cerebrovascular diseases (invention 4). a preparation containing greater burdock and truffles reduces both blood pressure and the level of blood fats (invention 19). a similar action is demonstrated by edible oils containing vegetable oils or animal fats as well as truffle-derived oils. the volume ratio of the abovementioned fats/oils in the raw material is 1:0.1–1 (invention 12). a preparation containing red tea and truffles also lowers blood fat levels (invention 24). of the above patented solutions, those involving cancers and tumors (patent 5), cardiovascular disease prevention and treatment (invention 4, 19) and immuno-resistance enhancement (invention 1–5, 7, 10–11, 19, 21–22, 24–26) should be highlighted for their applicability merits (practicability in use). the solutions are technologically quite simple and are easy to implement. a substantial disadvantage is that the raw material is expensive and, thus, highly reduces chances for implementation unless biotechnological processes are rolled out and put to use. acknowledgments this work was partially financed by mnisw under the initiative of “grants for grants” – support for polish coordinators in the research programs of the european union on the preparation of the project: “technology utilization in bio-geo environmental research: exemplary for innovative truffle biotechnology, domestication, economy and remote sensing (tuber)”, agreement no. 2515/gg 7. pr ue/2012/0 of 31.10.2012. authors would like to express sincere thanks to slobodan bajagić (croatia) for general idea for tuber project. first author would like to thanks mariusz grzesiczak (poland) for help in searching patent databases. authors’ contributions the following declarations about authors’ contributions to the research have been made: patent databases research and analysis, draft of the manuscript: mg; analysis of therapeutic potential of truffles inventions: fr; characterization of truffles properties: jg; final writing of the manuscript: mg, fr, jg. references 1. intellectual property statistics data center [internet]. 2014 [cited 2014 feb 20]; available from: http:// ipstatsdb.wipo.org/ipstatv2/ipstats/patentssearch 2. gajos m, hilszczańska d. research on truffles: scientific journals analysis. 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http://dx.doi.org/10.3923/pjbs.1999.510.514 http://dx.doi.org/10.1016/j.foodchem.2006.04.014 http://dx.doi.org/10.1002/ptr.2698 http://dx.doi.org/10.1016/j.biortech.2008.02.006 http://dx.doi.org/10.2174/1871520611009050424 http://dx.doi.org/10.2174/0929866511320040007 http://dx.doi.org/10.1016/s0039-128x(01)00105-2 http://dx.doi.org/10.1038/nature08867 http://dx.doi.org/10.1038/nature08867 http://dx.doi.org/10.1016/j.fgb.2010.10.007 http://dx.doi.org/10.1016/j.fgb.2010.11.003 http://dx.doi.org/10.1016/j.fgb.2010.11.003 http://dx.doi.org/10.1094/mpmi-07-11-0190 http://dx.doi.org/10.1016/j.foodchem.2011.11.077 318© the author(s) 2014 published by polish botanical society acta mycol 49(2):305–318 gajos et al. / therapeutic potential of truffles 26. white mj. esp@cenet europe’s network of patent databases. issues sci technol lib. 2006;47. http:// dx.doi.org/10.5062/f4f47m2h 27. gajos m. opis patenowy jako źródło informacji. katowice: university of silesia; 2000. 28. handbook on industrial property information and documentation [internet]. 2013 [cited 2013 dec 19]; available from: http://www.wipo.int/standards/en/part_03_standards.html 29. international patent classification – version 2013. guide [internet]. 2013 [cited 2013 dec 15]; available at: http://www.iponz.govt.nz/cms/iponz/latest-news/expired-items/international-patent-classification-2013 http://dx.doi.org/10.5062/f4f47m2h http://dx.doi.org/10.5062/f4f47m2h http://www.wipo.int/standards/en/part_03_standards.html http://www.iponz.govt.nz/cms/iponz/latest-news/expired-items/international-patent-classification-2013 abstract introduction material and methods results and discussion anticancer properties (invention 5) cardiovascular effect (invention 4, 19) reduction in blood fats (invention 12, 19, 24) immunological resistance (invention 1-5, 7, 10-11, 19, 21-22, 24-26) vigour growth (invention 1, 9, 20, 25) anti-aging benefits (invention 10, 11, 27) other conclusions acknowledgments authors’ contributions references 2014-12-31t17:48:52+0000 polish botanical society 2014-09-03t19:46:17+0200 polish botanical society 2014-08-28t17:49:05+0200 polish botanical society 2014-08-31t21:56:24+0200 polish botanical society 2014-08-31t21:55:48+0200 polish botanical society 2014-08-27t16:36:43+0200 polish botanical society 2014-08-27t17:38:15+0200 polish botanical society 2014-08-28t14:43:46+0200 polish botanical society 2014-09-02t20:10:40+0200 polish botanical society 2014-08-31t21:54:38+0200 polish botanical society 2014-08-27t16:49:45+0200 polish botanical society 2014-08-29t18:50:27+0200 polish botanical society 2014-09-02t20:09:46+0200 polish botanical society 2014-08-28t17:48:42+0200 polish botanical society 2014-09-02t20:10:59+0200 polish botanical society 2014-08-29t18:50:57+0200 polish botanical society 2014-08-31t21:57:05+0200 polish botanical society 2014-08-29t18:49:47+0200 polish botanical society 2014-08-31t21:56:10+0200 polish botanical society 2014-08-31t21:53:02+0200 polish botanical society 2014-09-06t20:24:58+0200 polish botanical society 2014-08-31t21:55:19+0200 polish botanical society 2014-09-02t20:10:16+0200 polish botanical society 2014-09-03t19:46:38+0200 polish botanical society 2014-08-31t21:54:58+0200 polish botanical society © the author(s) 2014 published by polish botanical society annotated checklist of fungi in cyprus island. 1. larger basidiomycota miguel torrejón la estrella, 18-1, e-12410, altura torrejon.miguel@gmail.com torrejón m.: annotated checklist of fungi in cyprus island. 1. larger basidiomycota. acta mycol. 49 (1): 109–134, 2014. an annotated checklist of wild fungi living in cyprus island has been compiled broughting together all the information collected from the different works dealing with fungi in this area throughout the three centuries of mycology in cyprus. this part contains 363 taxa of macroscopic basidiomycota. key words: macromycetes, inventory, ecology, distribution introduction in the first record, chatin (1897) dealt with terfezia aphroditis. nattrass (1937) in the most important work ever done in cyprus, were documented 92 species of anamorphic fungi, 104 species of ascomycetes, 148 species of basidiomycetes and 22 species of zygomycetes; nattrass & papaioannou (1938) added 20 species more. the same year dearness (1938), in a note, dealt with the species from the work of nattrass (1937), but there were no news. altson (1956), in a research into the cause of a rootrot illness on vicia faba, contributed with one species. the following year, 44 samples more were studied in georghiou & papadopoullos (1957). sixteen years later, all the previous information was compiled in the work of zyngas (1973) in which, 102 new references to the island were reported by the author. ten years later willoughby (1983) dealt with rhizophlyctis rosea. in the new millennium, winey (2005) in an illustrated book about macromycetes described 10 ascomycetes and 180 basidiomycetes to the island. in the next years, most authors dealt with single species originating from cyprus: tsopelas & nicolau (2005) reported heterobasidion annosum for the first time and tsopelas et al. (2008) researched seiridium cardinale and neophytou & ioannou (2009) recorded graphiola phoenicis. loizides (2008) dealt with macrofungi: five species of ascomycetes and twenty-seven basidiomycetes. momany & gücel (2009) in an illustrated guide gave information about 105 different species acta mycologica vol. 49 (1): 109–134 2014 doi: 10.5586/am.2014.011 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 110 m. torrejón © the author(s) 2014 published by polish botanical society of macromycetes. the same year in momany et al. (2009) nine species of agaricus were reported, but there is no new information in relation to the preceding work. also, edible and toxic fungi of cyprus, by loizides et al. (2011) is an important contribution improving the knowledge of macromycetes, in which eleven species of ascomycetes and eighty eight species of basidiomycetes were treated. besides, loizides (2008), loizides & kyriakou (2011) and loizides et al. (2012) concentrated on macromycetes. finally, torrejón (2013) added 25 species of anamorphic fungi, 28 species of ascomycetes, and 26 species of basidiomycetes to the island as you can see in table 1. table 1 published data on fungi in cyprus island authors’ names of the works dealing with fungi in cyprus number of anamorphic number of ascomycetes number of basidiomycetes number of zygomycetes chatin (1897) 1 nattrass (1937) 92 104 148 22 nattrass & papaioannou (1938) 2 5 8 5 altson (1956) 1 georghiou & papadopoullos (1957) 4 14 24 2 zyngas (1973) 26 62 4 10 willoughby (1983) 1 winey (2005) 10 180 tspelas & nicolau (2005) 1 tspelas et al. (2008) 1 loizides (2008) 5 27 neophytou & ioannou (2009) 1 momany & gücel (2009) 2 103 loizides et al. (2011) 11 88 loizides & kyriakou (2011) 1 14 loizides et al. (2012) 4 torrejón (2013) 25 28 26 total no. species 151 248 624 39 materials and methods all the previous information dealing with fungi in cyprus island were compiled. the different collections were disposed chronologically in each single species including the name of the species and the authors’ epithets, substrate/habitat, references and notes if necessary. duplicated species from two or more authors were grouped in a single one. results and discussion 138 species of anamorphs, 195 species of ascomycetes, 543 species of basidomycetes and 33 species of zygomycetes have been arranged alphabetically in chronological order. for further information see table 2. annotated checklist of fungi in cyprus island 111 © the author(s) 2014 published by polish botanical society table 2 numbers of fungal species listed in cyprus island number of anamorphic number of ascomycetes number of basidiomycetes number of zygomycetes total species from authors’ works dealing with fungi in cyprus 151 248 624 39 number of the same species from two or more works 13 53 81 6 total species in cyprus island 138 195 543 33 the summarized list contains 909 taxa and this causes the necessity of publishing the major part in the next issue(s). the present part contains the survey of larger basidiomycota (363 taxa). species have been arranged alphabetically and latin names and authors’ epithets according to index fungorum. agaricus aridicola geml, geiser & royse [=gyrophragmium dunalii (fr.) zeller] on dunes, girne, jan. 2005.wynei (2005). agaricus augustus fr. in black and calabrian pine forest, early autumn. loizides et al. (2008, 2011). uncommon. agaricus bernardii quél. on dunes and roadsides in land, alevkaya, autumn and winter. momany & gücel (2009). agaricus bisporus (j.e. lange) imbach in pastures, roadsides and cypress groves, autumn and winter. loizides et al. (2011). locally abundant. agaricus bitorquis (quél.) sacc. found in a stone quarry path, beşparmark, mar. 2000. in gardens, fields, parks and roadsides, autumn. wynei (2005); loizides et al. (2011). rare. agaricus bresadolanus bohus among grass in woodland or gardens, alevkaya, mar. momany & gücel (2009). agaricus campestris l. on grassy fields and newly dug garden soil, karaman, feb. 1997. on litter and pasture, alevkaya, winter and spring. in filds, pastures and cultivated land, winter and spring. wynei (2005); loizides (2008); momany & gücel (2009); loizides et al. (2011). frequent, but its numbers seem to be diminishing, probably due to the loss of its habitat. agaricus cappellianus hlaváček (as agaricus vaporarius (vittad.) m. m. moser) on pasture and lawns and in deciduous woods, alevkaya, autumn. momany & gücel (2009). agaricus gennadii (chatin & boud.) p. d. orton under eucalyptus sp., köprülü reservoir, dec. 2000. wynei (2005). agaricus impudicus (rea) pilát in all kind of forests, autumn. loizides et al. (2011). common. agaricus iodosmus heinem. in wood edges, parks and gardens, early in the autumn. loizides et al. (2011). rare. 112 m. torrejón © the author(s) 2014 published by polish botanical society agaricus moelleri wasser in all kind of forests, autumn and winter. loizides et al. (2011). occasional. agaricus placomyces peck on pastures and in mixed woodland on calcareous soils, alevkaya, winter. momany & gücel (2009). agaricus porphyrizon p. d. orton at the edge of a pinewood, beşparmak, dec. 2001. wynei (2005). agaricus porphyrocephalus f. h. møller on pastures and in mixed woodland, alevkaya, winter. momany & gücel (2009). agaricus pseudopratensis (bohus) waser in filds, roadsides, wood edges and cypress groves, autumn and winter. loizides et al. (2011). frequent. agaricus spp. without data. loizides (2008). agaricus sylvaticus schaeff. in mixed forests under pine trees, alevkaya, autumn. in all kinds of forests, autumn and spring. momany & gücel (2009). agaricus sylvicola (vittad.) peck. amongst grass in mixed forests, alevkaya, autumn. momany & gücel (2009); loizides et al. (2011.) agaricus xanthodermus genev. in gardens, parks, wood edges, roadsides and cypress groves, between autumn and spring. loizides et al. (2011). appearing sparingly. agaricus aff. macrocarpus on turf and pine litter, yaila, jan. 2001. wynei 2005. the only reason for qualifying the name with “aff.” is that the original description of the species indicates of coarse thick scales on the underside of the ring, which were no apparent in this collection. agaricus aff. sylvaticus without data. wynei (2005). agrocybe cylindracea (dc.) maire on broadleaved logs and trunks (without date). loizides et al. (2011). rare. agrocybe praecox (pers.) fayod amongst grasses in mixed forests, alevkaya, spring. momany & gücel (2009). amanita cistetorum contu & pacioni it grows exclusively with cistus, mainly c. salviifolius and c. monspeliensis, often very deeply rooted in the soil. loizides & kyriakou (2011). amanita codinae (maire) bertault on sandy coastal soil, the cengizköy golf-course, jan. 2001. wynei (2005). amanita ovoidea (bull.) link under pinus halepensis, ktima, dec. 1932. under or near pines, kalkanli, nov. 1999. in black and calabrian pine forests, as well as aleppo oak stands, autumn and winter. nattrass (1937); wynei (2005). loizides et al. (2011). as amanita ovoidea bull. in the first reference. amanita pantherina (dc.) krombh. in black pine forests, in early autumn. loizides et al. (2011). rare. amanita phalloides (vaill. ex fr.) link under the bark of populus nigra, lefkara, sep. 1933. wynei (2005); loizides (2008). annotated checklist of fungi in cyprus island 113 © the author(s) 2014 published by polish botanical society amanita proxima dumée on a gravelly roadside, alevkaya, nov. 1997. in black and calabrian pine forests, as well as aleppo oak stands, autumn and winter. wynei (2005); loizides et al. (2008, 2011). amanita torrendii justo [=torrendia pulchella bres.] under cistus as well as other mediterranean maquis (without date). loizides & kyriakou (2011). rare. amanita verna (bull.) lam. alevkaya forests, spring. momany & gücel (2009). amanita virosa (fr.) bertill. on soil surface in mixed or deciduous woodland, under a stone near boğaz water pump, apr. loizides (2008); momany & gücel (2009). amanita aff. malleata under holly-leaved oak, nov. 2000. wynei (2005). ampulloclitocybe clavipes (pers.) redhead, lutzoni, moncalvo & vilgalys [=clitocybe clavipes (pers.) p. kumm.] in mixed forests especially with coniferous trees, alevkaya, autumn. momany & gücel (2009). uncommon. antrodia serialis (fr.) donk it spreads in vertical patches along the fissures of pine bark, karaman, dec. 1999. wynei (2005). armillaria mellea (vahl) p. kumm. [=armillariella mellea (vahl) p. karst.] on roots and collar of citrus sinensis, famagusta; on morus alba, lapithos; on salix alba, kythrea. without data. on corylus avellana (without date and site). in a garden, growing from the underground stump of a falled tree, probly a plum, karaman, dec. 1999. on various broadleaved trees, autumn and winter. nattrass (1937); zyngas (1973 from georghiou & papadopoulos 1957); wynei (2005); loizides et al. (2011). frequent. as armillariella mellea (vahl) p. karst. in the second reference. armillaria obscura (schaeff.) herink [=armillaria polymyces (pers.) singer & clémençon] under ficus trees, alevkaya-aldalya, spring. momany & gücel (2009). astraeus hygrometricus (pers.) morgan on soil surface in mixed forests, alevkaya, mar. in cistus maquis, winter and spring. momany & gücel (2009); loizides & kyriakou (2011). auricularia auricula-judae (bull.) quél. on dead wood of citrus limonium, kythrea, oct. 1933; famagusta, oct. 1933; on dead wood of juglans regia, ktima, oct. 1933. on a falled mulberry, karaman, feb. 1998. on branches and logs of various broadleaved trees, from autumn to spring. nattrass (1937); wynei (2005); loizides et al. (2011). in the first references as auricularia auricula-judae (linn.) schröt. common. battarrea guicciardiniana ces. under eucalyptus sp., nicosia, nov. 1933. nattrass (1937). battarrea phalloides (dicks.) pers. amongst lawns and under tree forests, alevkaya, spring. momany & gücel (2009). battarrea stevenii (libosch.) fr. under olives, beyköy, (without date). wynei (2005). abundant in this habitat. 114 m. torrejón © the author(s) 2014 published by polish botanical society bjerkandera adusta (willd.) p. karst. on the concrete walls of a manhole, iskele, (without date). wynei (2005). boletus aereus bull. in black and calabrian pine forests, golden oak forests and mediterranean maquis, autumn. loizides et al. (2011). rare. boletus boudieri quél. under pinus halepensis, kornos, oct. 1931. nattrass (1937). boletus legaliae pilát under oak, early in autumn. loizides et al. (2011). boletus reticulatus schaeff. (as boletus reticulatus boud.) in black and calabrian pine forests, autumn. loizides et al. (2011). rare. boletus rhodopurpureus smotl. in a coniferous area, under an isolated pair of broadleaved trees (carob and hollyleaved oak), esentepe, nov. 1997. wynei (2005). boletus rhodoxanthus (krombh.) kallenb. in black and calabrian pine forests, as well as golden oak maquis, autumn. loizides et al. (2011). common in some years. boletus subtomentosus l. [=xerocomus subtomentosus (l.) quél.] in black and calabrian pine forests, autumn. loizides et al. (2011). bovista nigrescens pers. on the turf at the golf course, cengizköy, 2001. wynei (2005). bovista plumbea pers. on the turf at the folg course, cengizköy, 2001. on lawns amongst short grass and pasture, karşiyaka village, mar. wynei (2005); momany & gücel (2009). byssomerulius corium (pers.) parmasto on cut wood and bark of carob, (without data). wynei (2005). cantharellus cibarius fr. in pine forest, alevkaya, autumn. momany & gücel (2009). cantharellus subpruinosus eyssart. & buyck in black and calabrian pine forests, in autumn and winter. loizides et al. (2011). chalciporus amarellus (quél.) bataille under pines, alevkaya, jan. 1998. wynei (2005). chlorophyllum molybdites (g. mey.) massee in gardens, parks, greenhouses, roadsides and fields. loizides et al. (2011). rare. chondrostereum purpureum (pers.) pouzar parasitic or saprotrophic on many trees, alevkaya, in the whole year. momany & gücel (2009). chroogomphus rutilus (schaeff.) o. k. mill. under pines, growing on short turf, alevkaya, jan. 1998. in pine forest, alevkaya, autumn. in black and calabrian pine forests, from autumn to spring. wynei (2005); momany & gücel (2009); loizides et al. (2011). common. clathrus ruber p. micheli ex pers. under quercus infectoria, platres, jun. 1931. under cistus creticus, alevkaya, spring. nattrass (1937); momany & gücel (2009). clavaria incarnata weinm. growing out of cypress debris, yayla, jan. 2001. wynei (2005). annotated checklist of fungi in cyprus island 115 © the author(s) 2014 published by polish botanical society clavulina coralloides (l.) j. schröt. [=clavulina cristata (holmsk.) j. schröt.] on pine litter, alevkaya, jan. 1998. wynei (2005). cleistocybe carneogrisea (malençon) wizzini [=hygrophorus carneogriseus malençon] only on cypress litter, alevkaya, jan. 2001. wynei (2005). clitocybe alexandri (gillet) gillet under coniferous or broadleaved trees, essentepe, dec. 1998. in calabrian pine forests, in late autumn and spring. wynei (2005); loizides et al. (2011). common. clitocybe augeana (mont.) sacc. growing on manure (without date). wynei (2005). clitocybe candicans (pers.) p. kumm. girne, feb. 2000 (without data of substrate). wynei (2005). clitocybe costata kühner & romagn. under pines, alevkaya-yayla, dec. 2000. wynei (2005). clitocybe diatreta (fr.) p. kumm. under pines, esenteppe, nov. 2000. wynei (2005). clitocybe ditopa (fr.) gillet growing in a little cluster under pines, alevkaya, nov. 2000. wynei (2005). clitocybe fragrans (with.) p. kumm. amongst grasses, mosses or leaf litter beneath deciduous trees and pine forest, alevkaya, mar. momany & gücel (2009). clitocybe houghtonii (w. phillips) dennis on grass in mixed woodland forest, alevkaya, mar. momany & gücel (2009). clitocybe obsoleta (batsch) quél. growing in “fairy rings” in moss under cistus bushes, alsancak, dec. 2001. wynei (2005). clitocybe odora (bull.) p. kumm. in calabrian pine forests, alluvial broadleaved forests and aleppo oak stands, autumn and winter. loizides et al. (2011). occasional. clitocybe rivulosa (pers.) p. kumm. in gardens, parks, roadsides and wood edges, autumn and winter. loizides et al. (2011). uncommon. clitocybe umbilicata (schaeff.) p. kumm. on moss under pines, greçitköy reserve, dec. 2001. wynei (2005). clitocybe vermicularis (fr.) quél. on pine litter, alevkaya, mar. 1997. wynei (2005). clitocybe spp. without data. loizides (2008). clitopilus prunulus (scop.) p. kumm. on grass near eucalyptus, kanliköy, dec. 1998. wynei (2005). coltricia perennis (l.) murrill amongst grass on heath and woodland, alevkaya, spring. momany & gücel (2009). conferticium ochraceum (fr.) hallenb. on decaying wood of pinus nigra subsp. palliata, troodos, 17 nov. 2011. torrejón (2013). conocybe apala (fr.) arnolds [=bolbitius tener (gray) berk.] on mature-rich grass beside eucalyptus sp., kanliköy, dec. 1998. wynei (2005). 116 m. torrejón © the author(s) 2014 published by polish botanical society conocybe filaris (fr.) kühner [=pholiotina filaris (fr.) singer] on moss in a sandstone valley, kalkanli, feb. 2000. wynei (2005). conocybe pulchella (velen.) hauskn. & svrček [=conocybe pseudopilosella kühner ex kühner & romagn.] among grass and lawns in woodland, alevkaya, spring. momany & gücel (2009). conocybe subovalis kühner & watling [=galera ovalis (fr.) gillet] on ground, nicosia, (without data). nattrass (1937). coprinellus aff. flocculosus [=coprinus aff. flocculosus] on the side of a tiny brook, kalavaç, apr. 2000. wynei (2005). coprinellus domesticus (bolton) vilgalys, hopple & jacq. johnson [=coprinus domesticus (bolton) gray] on buried timber, nov. 1997. wynei (2005). coprinopsis atramentaria (bull.) redhead, vilgalys & moncalvo [=coprinus atramentarius (bull.) fr.] on ground, nicosia, mar. 1932. nattrass (1937). coprinopsis cinerea (schaeff.) redhead, vilgalys & moncalvo [=coprinus cinereus (schaeff.) gray] growing in a large numbers mixed with another smaller fungus on a sheep midden. wynei (2005). coprinopsis cothurnata (godey) redhead, vilgalys & moncalvo [=coprinus cothurnatus godey] in a midden-heap on a sheep dropping, yeniceköy, jan. 1998. wynei (2005). coprinopsis picacea (bull.) redhead, vilgalys & moncalvo [=coprinus picaceus (bull.) gray] mixed woods, alevkaya, jan. 2000; under pistacia terebinthus, küçükerenköy, jan. 2000. on soil surface in mixed forests, karşiyaka village, apr. wynei (2005); momany & gücel (2009). coprinopsis romagnesiana (singer) redhead, vilgalys & moncalvo on or around rotten wood (without date and site). loizides et al. (2011). uncommon. coprinus comatus (o. f. müll.) pers. on a rubbish dump, esentepe, dec. 2000; on ploughed soil, at ağillar, feb. 2001. without data. in grass by roadsides, rubbish heaps or lawns and under trees on remnants of fallen leaves, girne-alsancak, feb. in parks, fields, wood edges and road edges, often on disturbed ground, autumn and spring. wynei (2005); loizides et al. (2008, 2011); momany & gücel (2009). as coprinus ovatus (schaeff.) fr. in the second habitat from the first reference. coprinus disseminatus (pers.) j. e. lange on stumps of broadleaved trees or on soil, karaoğlanoğlu, mar. momany & gücel (2009). coprinus spp. without data. loizides (2008). coriolellus albidus (fr.) bondartsev on fallen or dead branches of deciduous trees, alevkaya, may. momany & gücel (2009). annotated checklist of fungi in cyprus island 117 © the author(s) 2014 published by polish botanical society coriolopsis gallica (fr.) ryvarden [=trametes hispida bagl.] on branch of prunus armeniaca, ay. amvrosios, kyrenia, jun.1931. on live walnut and on cut timber, (without site and date). nattrass (1937); wynei (2005). coriolus hirsutus (wulfen) quél. on dead wood and fallen trunks of deciduous trees, alevkaya, may. momany & gücel (2009). cortinarius caligatus malençon in calabrian pine forests and mediterranean maquis in association with rockrose cistus sp. in cistus maquis, (without date). loizides et al. (2011); loizides & kyriakou (2011). cortinarius collinitus (pers.) fr. on soil in mixed forests and thorny shrubs, more rarely in deciduous, gecitköy, mar. momany & gücel (2009). cortinarius dionysae rob. henry in cistus garigue, beşparmak, dec. 2000. wynei (2005). cortinarius juranus remaux in calabrian pine and golden oak forests, autumn and winter. loizides et al. (2011). rare. cortinarius mucosus (bull.) j. kickx in conifer heath, usually with pine, alevkaya, autumn. momany & gücel (2009). cortinarius splendens rob. henry in calabrian pine and golden oak forests, autumn and winter. loizides et al. (2011). cortinarius sp. on burn ground in cistus and lentisk garigue, buffavento track, jan. 2001. wynei (2005). crepidotus cesatii (rabenh.) sacc. on dead twigs of dittrichia viscosa, özhan, dec. 1998. wynei (2005). crepidotus mollis (schaeff.) staude on a carob, tazik kiran woods, dec. 1998. wynei (2005). crepidotus variabilis (pers.) p. kumm. on terebinth twigs, jan. 1998. wynei (2005). crinipellis scabella (alb. & schwein.) murrill on dead grass stems, gönyeli, nov. 1997. wynei (2005). cryptosporiopsis corticola (edgerton) nannf. [=mysosporium corticolum edgerton] on dead twigs and cankers on branch of pyrus malus, prodhromos, oct. 1931. nattrass (1937). cyathus olla (batsch) pers. on a heap of plant remains, kanliköy, dec. 1998. wynei (2005). cystoderma terreyi (berk. & broome) harmaja between alevkaya and yayla, dec. 1998. wynei (2005). cystoderna amianthinun (scop.) fayod in pineland, alevkaya, jan. 1998. wynei (2005). frequent. cystolepiota adulterina (f. h. møller) bon [=lepiota adulterina f. h. møller] amongst broadleaved woods as nettle and scrub, alevkaya, autumn. momany & gücel (2009). cystolepiota cystophora (malençon) bon on stony ground under carobs, alevkaya, ja. 1999. wynei (2005). 118 m. torrejón © the author(s) 2014 published by polish botanical society cystolepiota moelleri knudsen [=lepiota rosea rea] on damp ground under deciduous trees, alevkaya, autumn. momany & gücel (2009). dacrymyces variisporus mcnaab on wood from branches of pinus nigra subsp. palliata, troodos, 17 nov. 2011. torrejón (2013). dacryobolus karstenii (bres.) oberw. ex parmasto [=stereum karstenii bres.] on wood of pinus halepensis, troödos, aug. 1931. nattrass (1937). entoloma hirtipes (schumach.) m. m. moser [=nolanea hirtipes (schumach.) p. kumm.] on grass or on rich humus in pine forests, alevkaya, mar. momany & gücel (2009). entoloma jubatum (fr.) p. karst. in pinewoods, alevkaya, jan. 1998. wynei (2005). frequent. entoloma lividoalbum (kühner & romagn.) kubička in calabrian pine and golden oak forests, autumn and winter. loizides et al. (2011). occasional. entoloma phaeocyathus noordel. in the cengizköy golf-course, dec. 2001. wynei (2005). entoloma aff. hirtipes in pinewoods, alevkaya (without date). wynei (2005). entoloma aff. undatum on mossy ground under pines, st. hilarion, dec. 1998. wynei (2005). exidia plana donk on dead deciduous wood, alevkaya, dec. 1998. wynei (2005). exidiopsis calcea (pers.) k.wells on wood from a branch of pinus brutia, trail to calydonian falls, 19 nov. 2011. torrejón (2013). exobasidium vaccinii (fuckel) woronin on leaves of arbutus andrachne troodos, jun. 1935. nattrass (1937). fomes fomentarius (l.) fr. on trunk of populus nigra, saitta, feb. 1932; of platanus orientalis, stavros, jan. 1936. on birch and on almond, alevkaya and pedoulas, mar. nattrass (1937); momany & gücel (2009). fomitiporia punctata (pilát) murrill [=poria friesiana bres.] [=phellinus punctatus pilat] on citrus sinensis, famafusta, jul. 1933. on olive stumps and roots, karaman, (without date). nattrass (1937); wynei (2005). fuscoporia torulosa (pers.) t. wagner & m. fisch. [=fomes torulosus (pers.) lloid] [=phellinus torulosus (pers.) bourdot & galzin] on trunk of ceratonia siliqua, lefkara, apr. 1931. attached to carob and almond trees, (without site and date). on tree stumps, carob tree stem, pomegranate, kasyake, lapta, alevkaya, mar. nattrass (1937); wynei (2005); momany & gücel (2009). galerina marginata (batsch) kühner beside a grassy firebreak, alevkaya, jan. 2002. on dead (sometimes buried) wood, autumn and winter. wynei (2005); loizides et al. (2008, 2011). frequent. annotated checklist of fungi in cyprus island 119 © the author(s) 2014 published by polish botanical society galerina pumila (pers.) m. lange [=galerina mycenopsis (fr.) kühner] on soil amongst moss in grassland, tepebaşi altkisimlari, mar. momany & gücel (2009). galerina vittiformis (fr.) singer on mossy soil among cistus, under pines, dec. 1999, (without site). wynei (2005). ganoderma adspersum (schulzer) donk round a fig stem and on the buried remains of a dead almond, karaman, (without date). wynei (2005); momany & gücel (2009). ganoderma applanatum (pers.) pat. on trunk of ceratonia siliqua, ay. theodhoros, larnaca, jun. 1931; on root of alnus orientalis, marathasa, mar. 1936. on citrus limonium, (without date and site). nattrass (1937); zyngas (1973 from nattrass & papaioannou 1938). ganoderma lucidum (curtis) p. karst. on ground, agros, oct. 1934. nattrass (1937). ganoderma resinaceum boud. on stumps of carobs, alevkaya, (without date). momany & gücel (2009). ganoderma sp. (as ganoderma solani (mart.) sacc.) on stem of ceratonia siliqua, akanthou, 1967. zyngas (1973).there is not a fungus with the name used by zyngas. it might be fuscoporia torulosa, the most common species living in this habitat in all mediterranean coastal area. geastrum berkeleyi massee on pine litter, mt. yayla, dec. 2001. wynei (2005). geastrum fimbriatum fr. [=geastrum sessile (sowerby) pouzar] on rich humus or pine needles in mixed forest, alevkaya, mar. momany & gücel (2009). geastrum rufescens pers. on soil surface in deciduous and coniferous woodland, alevkaya, spring. momany & gücel (2009). geastrum triplex jungh. on leaf litter in deciduous wood, parks and gardens and mainly under pine trees, alevkaya, feb. momany & gücel (2009); wynei (2005). geastrum pseudolimbatum hollós on a bare patch of sandy soil, cengizköy golf course, dec. 2011. wynei (2005). gloeophyllum abietinum (bull.) p. karst. [=lenzites abietina (bull.) fr.] on worked timber, nicosia, may 1931. on the sides and cut surfaces of falled cypresses and pines (without site and date). on wood tree stumps or durned forests, alevkaya (without data). nattrass (1937); wynei (2005); momany & gücel (2009). as lenzites abietina (bull.) fr. in the first reference. gloeophyllum trabeum (pers.) murrill [=lenzites trabea (pers.) fr.] on trunk of pinus halepensis, stavros, sep. 1935. on dead conifers and sawn wood, (without site and data). nattrass (1937); wynei (2005). grifola frondosa (dicks.) gray on an indeterminate tree, yenierenköy, (without date). wynei (2005). gymnopilus flavus (bres.) singer on soil with wood chippings, near cengizköy paper mill, dec. 2001. wynei (2005). gymnopilus junonius (fr.) p.d. orton on dead wood, autumn and winter. loizides et al. (2011). rare. 120 m. torrejón © the author(s) 2014 published by polish botanical society gymnopus dryophilus (bull.) murrill [=collybia dryophila (bull.) p. kumm.] on moss or grass under trees, from oct., (without site). wynei (2005). gymnopus peronatus (bolton) gray [=collybia peronata (bolton) p. kumm.] on leaf litter in mixed woodland forests, alevkaya, spring. momany & gücel (2009). hebeloma cistophilum maire beneath cistus salvifolius, c. albidus, c. monspeliensis and c. laurifolius, autumn and winter. loizides & kyriakou (2011). hebeloma sinapizans (paulet) gillet (as hebeloma sinapizans (pers.) fr.) in black and calabrian pine forests as well as golden oak forests, autumn and winter. loizides et al. (2011). common. heterobasidion annosum (fr.) bref. on stumps of pinus nigra ssp. pallasiana,troodos, jun. 2003. tsopelas & nikolaou (2005). hohembuehelia geogenia (dc. ex fr.) singer on buried wood from olive debris, alevkaya, jan. 2001. wynei (2005). hydnum rufescens pers. in black and calabrian pine forests, golden oak forests and alluvial broadleaved forests. loizides et al. (2011). rare. hygrocybe acutoconica (clemen.) singer [=hygrocybe langei kühner] amongs lawns and pasture, alevkaya, autumn. momany & gücel (2009). hygrocybe chlorophana (fr.) wünsche on grassy patches, esentepe and bahçeli, 1999. wynei (2005). hygrocybe conica (schaeff.) p. kumm. on lawn, alevkaya, feb. 1997; lefkoça, jan. 2001. wynei (2005). hygrocybe virginea (wulfen) p.d. orton & watling [=cuphophyllus virgineus (wulfen) kovalenko] in shady places under pines, esentepe, (without date). wynei (2005). hygrophorus chrysodon (batsch) fr. in cistus maquis, (without date). loizides & kyriakou (2011). hygrophorus cossus (sowerby) fr. amongst grass in mixed forests, alevkaya, autumn. momany & gücel (2009). hygrophorus hypothejus (fr.) fr. in calabrian pine forests, winter. loizides et al. (2011). common. hygrophorus latitabundus britzelm. in a pinewood, esentepe, dec. 2000. in black pine forests, autumn. wynei (2005); loizides et al. (2011). locally abundant, but not very widespread. hygrophorus pseudodiscoideus var. cistophilus bon & g. riousset under cistus salviifolius and c. monspeliensis, (without date). loizides & kyriakou (2011). hygrophorus pustulatus (pers.) fr. amongst grass in mixed forest especially with pine, alevkaya, spring. momany & gücel (2009). hygrophorus spp. without data. loizides (2008). hyphoderma incrustatum k. h. larss. on decaying wood from a log of pinus nigra subsp. palliata, troodos, 17 nov. 2011. torrejón (2013). annotated checklist of fungi in cyprus island 121 © the author(s) 2014 published by polish botanical society hyphoderma nemorale k. h. larss. on decaying wood from a branch of quercus alnifolia, trail to calydonian falls, 19 nov. 2011.torrejón (2013). hyphodontia sambuci (pers.) j. erikss. on dead deciduous wood, alevkaya, mar. momany & gücel (2009). hypholoma capnoides (fr.) p. kumm. on stumps of unknown trees, alevkaya, spring. momany & gücel (2009). hypholoma fasciculare (huds.) p. kumm. on dead or buried wood, autumn and spring. loizides et al. (2011). hypholoma marginatum j. schröt. on soil under conifers, karaoğlanoğlu, mar. momany & gücel (2009). infundibulicybe geotropa (bull.) harmaja in calabrian pine forest, aleppo oak stands and alluvial broadleaved forests, autumn and winter. loizides et al. (2011). inocybe bongardii var. pisciodora (donadini & riousset) kuyper [=inocybe pisciodora donadini & riousset] on turf under cistus, tarzik viran, feb. 2000. wynei (2005). numerous in this habitat. inocybe cincinnata (fr.) quél. amongst grass in deciduous and coniferous woodlands, alevkaya, spring. momany & gücel (2009). inocybe dulcamara (pers.) p. kumm. small troops on turf under cistus and at wood edges, karaman, may 2000. on dunes or lawns, alevkaya, winter. wynei (2005); momany & gücel (2009). inocybe flocculosa (berk.) sacc. [=inocybe flocculosa var. crocifolia (herink) kuiper] on a sandstone valley soil, under cistus and inula shrublets, kalkanli, feb. 2000; under pines, alevkaya, jan. 2002. on soil surface in mixed forests, akdeniz drmanlari, mar. inside or outside forests, often on calcareous soil, autum, and winter and spring. in cistus maquis, (without date). wynei (2005); momany & gücel (2009); loizides et al. (2011); loizides & kyriakou (2011). common. as inocybe flocculosa var. crocifolia in the first reference. inocybe fraudans (britzelm.) sacc. in black and calabrian pine forests, autumn and winter. loizides et al. (2011). inocybe geophylla (fr.) p. kumm. in black and calabrian pine forests, cedar forests and alluvial broadleaved forests, autumn, winter and occasionally in spring. loizides et al. (2011). common. inocybe godeyi gillet in deciduous woodland and mixed forests, alevkaya, autumn and spring. momany & gücel (2009). inocybe griseolilacina j. e. lange deciduous trees and roadsides, alevkaya, winter. momany & gücel (2009). inocybe griseovelata kühner in pinewoods, esentepe, dec. 1999. wynei (2005). inocybe hirtella bres. in parks and deciduous forests, particularly under beech and lime trees on calcareous humus-rich soils, alevkaya, mar. momany & gücel (2009). 122 m. torrejón © the author(s) 2014 published by polish botanical society inocybe lacera (fr.) p. kumm. in pine woods, alevkaya, feb. 1998. under pine trees, alevkaya, winter. wynei (2005); momany & gücel (2009). inocybe obscurobadia (j. favre) grund & d. e. stuntz under pines, anthos, camlibel, jan. 2000. wynei (2005). inocybe petiginosa (fr.) gillet deciduous woodland, tepebasi-altkisimlari, mar. momany & gücel (2009). inocybe rimosa (bull.) p. kumm. mainly in black pine forests, less often in calabrian pine forests, early in the autumn. loizides et al. (2011). very common. inocybe aff. bongardii on pine litter, alevkaya, feb. 1999. wynei (2005). inocybe aff. nitidiuscula on moss under pines, alevkaya, feb. 1997. wynei (2005). inocybe aff. sindonia on pine litter, alevkaya, jan. 2002. wynei (2005). inocybe spp. without data. loizides (2008). inonotus hispidus (bull.) p. karst. [=polyporus hispidus (bull.) fr.] on trunk of populus nigra, nicosia, nov. 1931. nattrass (1937). lacrymaria lacrymabunda (bull.) pat. [=psathyrella lacrymabunda (bull.) m. m. moser] on soil and pine debris in a shaded courtyard, karaman, dec. 2000. wynei (2005). lactarius chrysorrheus fr. under golden oak, autumn and winter. loizides et al. (2011). common. lactarius deliciosus (l.) gray under pinus halepensis, troodos, (without date). under pines and among terevinth shrubs, between alevkaya and the beşparmak hills, (without date). amongst grass under pines or spruce trees, alevkaya, autumn and spring. in black and calabrian pine forests, autumn and winter. nattrass (1937); wynei (2005); loizides (2008); loizides et al. (2011); momany & gücel (2009). as lactarius deliciosus (linn.) fr. in the first reference. frequent. lactarius hepaticus plowr. on litter in mixed forests especially with pine, alevkaya, spring. momany & gücel (2009). lactarius hysginus (fr.) fr. in mixed forests, alevkaya, winter. momany & gücel (2009). lactarius mammosus fr. in mixed forests especially under pine trees, gecitköy, mar. momany & gücel (2009). lactarius sanguifluus (paulet) fr. mainly in black pine forests in altitudes over 1.200 m. loizides et al. (2011). common. lactarius semisanguifluus r. heim & leclair in calabrian pine forests, often in calcareous soil, autumn and winter. loizides et al. (2011). common. annotated checklist of fungi in cyprus island 123 © the author(s) 2014 published by polish botanical society lactarius tesquorum malençon in symbiosis with rockrose cistus spp., autumn, winter and spring. in cistus maquis, (without date). loizides et al. (2011); loizides & kyriakou (2011). common. one of the commonest cistus-specific species. lactarius volemus (fr.) fr. under trees in mixed forests, alevkaya, winter. momany & gücel (2009). lactarius spp. without data. loizides (2008). laetiporus sulfureus (bull.) murrill [=polyporus sulphureus (bull.) fr.] on ceratonia siliqua (without date and site). kicked down on melia azedarach, girne, nov. 1999. on dead or alive broadleaved trees, autumn and spring. zyngas (1973 from nattrass & papaioannou 1938); wynei (2005); loizides et al. (2011). as polyporus sulphureus (bull.) fr. in the first reference. leccinellum corsicum (rolland) bresinsky & manfr. binder associated with cistus monspeliensis, c. ladanifer, c. incanus and c. salviifolius, (without date). loizides & kyriakou (2011). leccinellum lepidum (h. bouchet ex essette) bresinsky & manfr. binder under golden oak and kermes oak, autumn and winter. loizides et al. (2011). frequent in some years. lepiota aspera (pers.) quél. in alluvial, broadleaved forests, parks, scrubland and roadsides, autumn. loizides et al. (2011). uncommon. lepiota brunneoincarnata chodat & c. martín without data. in forests, parks, fields, roadsides and gardens, autumn and winter. loizides (2008); loizides et al. (2011). occasional. lepiota castanea quél. in calabrian pine forests, alluvial broadleaved forests, wood edges and scrubland, autumn and winter. wynei (2005); loizides et al. (2011). frequent. lepiota pseudolilacea huijsman in calabrian pine forests, roadsides, cypress groves, wood edges and scrubland, autumn and winter. loizides et al. (2011). occasional. lepiota subincarnata j. e. lange in calabrian pine forests, alluvial broadleaved forests, wood edges, paarks and gardens, often among grass, autumn and winter. loizides et al. (2011). occasional. lepiota aff. bruneoincarnata on grassy bank, gecitköy, dec. 2001. wynei (2005). lepiota aff. clypeolaria on mossy turf, alevkaya, jan. 2002.wynei (2005). lepista flaccida (sowerby) pat. [=clitocybe inversa (scop.) quél.] on moss under pines, geçitköy reserve, dec. 2001. wynei (2005). lepista nuda (bull.) cooke around girne and alevkaya, jan. and feb. (without year). in all kind of forests, autumn and winter. wynei (2005); loizides (2008); loizides et al. (2011).quite frequent. lepista sordida (schumach.) singer without data. loizides (2008). 124 m. torrejón © the author(s) 2014 published by polish botanical society leucoagaricus leucothites (vittad.) wasser numerous on turf, buffavento road, nov. 2000. in parks, gardens, road edges and round-about among grass, autumn and winter. wynei (2005); loizides et al. (2011). leucoagaricus serenus (fr.) bon & boiffard [=sericeomyces serenus (fr.) heinem.] under pines, cengizköy, nov. 1998. wynei (2005). limacella illinita (fr.) maire in pine woods, st. hilarion, dec. 2000. wynei (2005). limacella subfurfuracea contu in pinewoods, alevkaya and akdeniz, dec. 1998, 2000. wynei (2005). lycoperdon excipuliforme (scop.) pers. in black and calabrian pine forests, as well as golden oak maquis, autumn and spring. loizides et al. (2011). frequent. lycoperdon nigrescens pers. [=lycoperdon foetidum bonord.] on soil in coniferous or mixed woodland, alevkaya, mar. momany & gücel (2009). lycoperdon perlatum pers. on soil under pines, alevkaya (without date). on soil surface in mixed woodland, alevkaya and prodromos forests, mar. in black and calabrian pine forests, autumn. wynei (2005); momany & gücel (2009); loizides et al. (2011). common. lycoperdon pratense pers. [=vascellum pratense (pers.) kreisel] on turf, dengizköy golf course, dec. 2001. on soil surface in lawns and pasture, karşiyaka village, apr. wynei (2005); momany & gücel (2009). lycoperdon pyriforme schaeff. on rotten logs or stumps or on soil surface but attached to buried wood by a mycelial cords, karşiyaka village, mar. momany & gücel (2009). lyophyllum connatum (schumach.) singer in black and calabrian pine and golden oak forests. loizides et al. (2011). occasional. lyophyllum decastes (fr.) singer in a garrigue, buffavento dirt road, jan. 2001. wynei (2005). lyophyllum fumosum (pers.) p. d. orton in black and calabrian pine and golden oak forests, autumn and winter. loizides et al. (2011). frequent in some years. lyophyllum sp. on the cengizköy golf course, dec. 2001. wynei (2005). macrolepiota excoriata (schaeff.) wasser in wood edges, fields, gardens and road edges, autumn and winter. loizides et al. (2011). macrolepiota konradii (huijsman ex p. d. orton) m. m. moser in a pinewood, alevkaya, nov. 2000. wynei (2005). marasmiellus ramealis (bull.) singer [=marasmius ramealis (bull.) fr.] on old wood material, tepebasi-altkisimlari, mar. momany & gücel (2009). marasmius anomalus peck on turf, gönyeli, jan. 1998. wynei (2005). marasmius rotula (scop.) fr. on turf, armenian monastery, nov. 2000. wynei (2005). marasmius wynneae berk. & broome (as marasmius wynnei) under pines, alevkaya, dec. 1998. wynei (2005). annotated checklist of fungi in cyprus island 125 © the author(s) 2014 published by polish botanical society melanoleuca brevipes (bull.) pat. on floor of gravel quarry, ridge road, (without date). wynei (2005). melanoleuca excissa (fr.) singer amongs clover on sandy soil, cengizköy golf course, dec. 2004. wynei (2005). melanoleuca paedida (fr.) künher & maire on moss under cistus bushes, arapköy, feb. 2001. wynei (2005). melanoleuca rasilis var. rasilis (fr.) singer alongside a broken log on dunes, girne, dec. 2004. wynei (2005). melanoleuca strictipes (p. karst.) jul. schäff. in woodland amongst grass or pasture, alevkaya, autumn. momany & gücel (2009). melanoleuca aff. excissa on bare soil, alevkaya, feb. 2000. wynei (2005). micromphale brassicolens (romafn.) p. d. orton on pine-litter, alevkaya, nov. 2000. wynei (2005). montagnea haussknechtii rabenh. on sand dunes, karpaz, may 1997. wynei (2005). mycena arcangeliana bres. [=mycena oortiana hora] on a dead mossy carob, tazik kiran, feb. 1999. on tree stumps and branches of deciduous trees, alevkaya, winter. wynei (2005); momany & gücel (2009). mycena amicta (fr.) quél. widespread on pine cones, küçükerenköy, jan. 2000. wynei (2005). mycena crocata (schrad.) p. kumm. on leaf litter in mixed forests, beşparmak, winter. momany & gücel (2009). mycena galericulata (scop.) gray on fallen branches and stumps of broadleaved trees, alevkaya, autumn. momany & gücel (2009). mycena galopus (pers.) p. kumm. on cistus debris and pine litter, (without site and date). amongst leaf litter in woodlands, alevkaya, spring. wynei (2005); momany & gücel (2009). fairly common. mycena megaspora kauffman [=mycena uracea a. pearson] on heath in grassland and mixed forests, akdeniz ormanlari, mar. momany & gücel (2009). mycena pelianthina (fr.) quél. on litter in mixed forests, alevkaya, spring.momany & gücel (2009). mycena pura (pers.) p. kumm. in pine woods, kozan, dec. 1998. in black, calabrian pines and cedar forests, as well as in alluvial broadleaved forests, autumn. wynei (2005). loizides et al. (2011). common. mycena sanguinolenta (alb. & schwein.) p. kumm. growing from a cracked carob stump, alevkaya, dec. 2001. wynei (2005). it was observed on soil and truf too. mycena seynesii quél. on pine cones on the ground or buried, dec.-jan. (without site). on pine cones and coniferous debris, alevkaya, winter. wynei (2005); momany & gücel (2009). common in pinewoods. mycena spp. without data. loizides (2008). 126 m. torrejón © the author(s) 2014 published by polish botanical society myriostoma coliforme (dicks.) corda two basidiomata, on soil very rich in humus beneath quercus alnifolia and platanus orientalis, close to cyclamen cyprius, cedar valley, 22 nov. 2011. torrejón (2013). rare. included by the european council for the conservation of fungi as a candidate to the list of threatened fungi in europe. myxomphalia maura (fr.) hora often in troops on burnt ground, as in garrigue, buffavento dirt road, jan. 2001. wynei (2005). omphallina sp. on a mossy boulder, karaman, dec. 1999. wynei (2005). omphaltus olearius (dc.) singer on olive trees, (without date and site). on olive and other broadleaved trees, sometimes at the base of trunks appearing terrestrial, but always associated with wood, (without date). wynei (2005); loizides (2008); loizides et al. (2011). panaeolina foenisecii (pers.) maire in grassland and on loans and roadsides, tepebasi altkisimlari, mar. momany & gücel (2009). panaeolus aff. olivaceus on burnt ground, karaman, dec. 1999. wynei (2005). this collection almost matches the descriptions of this rare european species which has somewhat broarder spores; hence the qualification “aff.”. panaeolus papilionaceus (bull.) quél. on a wayside grass, atifonitis monastery, dec. 2000. wynei (2005). panus conchatus (bull.) fr. [=lentinus conchatus (bull.) j. schröt] on soil between rocks, but perhaps attached to olive or carob roots, karşiyaka, jan. 2001. wynei (2005). parasola megasperma (p. d. orton) redhead, vilgalys & hopple [=coprinus megaspermus p. d. orton] on the ground, lapta, dec. 2004. wynei (2005). parasola plicatilis (curtis) redhead, vilgalys & hopple [=coprinus plicatilis (curtis) fr.] on turf and shady wood sides, alevkaya, jan. 2001. wynei (2005). paxillus rubicundulus p.d. orton in mycorrhizal symbiosis with alder, autumn and spring. loizides et al. (2011). peniophora cinerea (pers.) cooke on a decaying branch of ceratonia siliqua, asgata i, 24 nov. 2011. torrejón (2013). peniophora lycii (pers.) höhn. & litsch. [=peniophora caesia bres.] on branch opf ceratonia siliqua, lefkara, jun. 1931. on a decaying branch of cistus creticus, asgata ii, 24 nov. 2011. nattrass (1937); torrejón (2013). as peniophora caesia bres. in the first reference. peniophora meridionalis boidin on a decaying branch of quercus infectoria, agia paraskevi, 23 nov. 2011. torrejón (2013). peniophora quercina (pers.) cooke on a decaying branch of quercus infectoria, agia paraskevi, 23 nov. 2011. torrejón (2013). annotated checklist of fungi in cyprus island 127 © the author(s) 2014 published by polish botanical society peniophora sp. on corylus avellana (without date and site). zyngas (1973 from georghiou & papadopoulos 1957). phaeolus schweinitzii (fr.) pat. [=polyporus schweinitzii fr.] on trunk of pinus halepensis, stavros, jan. 1936. nattrass (1937). phallus impudicus l. under cistus sp. karaman, jan. 2001. wynei (2005). phellinus igniarius (l.) quél. on tamarisk, ortaköy, jan. 2001. wynei (2005). phellinus pomaceus (pers.) maire [=fomes pomaceus (pers.) lloyd] on trunk of prunus persica, pedhoulas, jun. 1931; on branch of prunus cerasus, prodhromos, oct. 1933; on branch of prunus amygdalus, agros, jan. 1936; on branch of prunus domestica, ay. amvrosios, kyrenia, feb. 1936; on trunk and branch of prunus armeniaca, limassol, feb. 1936. nattrass (1937). phellinus rimosus (berk.) pilát on a tree-size terebinth, karaağaç. wynei (2005). phellodon tomentosus (l.) banker coniferous and mixed forests, alevkaya, autumn and spring. momany & gücel (2009). phoeomana speirea (fr.) redhead [=mycena speirea (fr.) gillet] on wood and twigs, alevkaya, mar. momany & gücel (2009). pholiota highlandensis (peck) quadr. & lunghini [=pholiota carbonaria (fr.) singer] on burnt ground, beşparmak peaks, dec.-jan. 1995-1996. on soil surface in burned forests, alevkaya, autumn. wynei (2005); momany & gücel (2009). many groups seen after the 1995 fire. pholiota mixta (fr.) kuyper & tjall.-beuk. on buried conifer wood, buffavento, dirt road, jan. 2001. wynei (2005). phylloporia ribis (schumach.) ryvarden [=fomes ribis (schumach.) gillet] on collar of rosa sp., nicosia, jul. 1931. parasitic on a branch of cistus sp., kayalar, dec. 2001. on rosa sp. (without date and site). nattrass (1937); wynei (2005); zyngas (1973 from nattrass & papaioannou 1938). as fomes ribis (schumach.) gillet in the first reference, and as phellinus ribis (schumach.) quél. in the last one. pisolithus arrhizus (scop.) rauschert under eucalyptus sp., vadili (without date). wynei (2005). widely scattered elsewhere under other trees. pleurotus eryngii (dc.) quél. on rich humus on the ground amongst moss, taşkent, pigades-hisarköy, geçitköy and tepebasi altkisivolari, mar. momany & gücel (2009). pleurotus eryngii var. ferulae (lanzi) sacc. [=pleurotus fuscus var. ferulae (lanzi) bres.] among and probably in association with ferula communis, salamis, 1931. dec.-apr., (without a specific site data). growing saprotrophically or parasitically on ferula communis, (without date). on giant fennel plants or remains, between autumn and spring. nattrass (1937); wynei (2005); loizides (2008); loizides et al. (2011). a much esteemed comestible and frequently offered for sale. as pleurotus fuscus var. ferulae (lanzi) bres. in the first reference. 128 m. torrejón © the author(s) 2014 published by polish botanical society pleurotus ostreatus (jacq.) p. kumm. on various broadleaved trees (whithout date). loizides et al. (2011). rare. pleurotus pulmonarius (fr.) quél. on deciduous wood or buried roots, tepebaşi, apr. momany & gücel (2009). pluteus aff. cinereofuscus in a damp site, karaman, nov. 1998. wynei (2005). polyporus meridionalis (a. david) h. jahn [=polyporus meridionalis (a. david) tellería] on fallen twigs of deciduous trees, karaoğlanoğlu, mar. on fallen cistus twigs and debris of other sclerophyllous plants, growing throughout the year. wynei (2005); momany & gücel (2009); loizides & kyriakou (2011). quite common on roots of pine trees and garigue shrubs. as polyporus meridionalis (a. david) tellería in the second reference. polysaccum crassipes dc. under eucalyptus sp., oct. 1932. nattrass (1937). porodaedalea pini (brot.) murrill [=trametes pini (brot.) fr.] on trunk of pinus halepensis, kyrenia, jun. 1931. nattrass (1937). psathyrella artemisiae (pass.) konrad & maubl. (as psathyrella squamosa (p. karst.) m. m. moser) in mixed woodland, alevkaya, autumn and spring. momany & gücel (2009). psathyrella bipellis (quél.) a. h. sm. on turf among dittrichia sp. shrublets in a damp valley, kalkanli, dec. 1999. wynei (2005). psathyrella candolleana (fr.) maire on turf around pinewoods, atifonitis monastery, feb. 2002. near deciduous trees, stumps or cut trees, girne-alsancak and karaoğlanoğlu, feb. wynei (2005); momany & gücel (2009). rather common. psathyrella sp.1. several troops on turf in olive valley, kalkanli, jan. 2001. wynei (2005). psathyrella sp.2. in troops on grassy bank, mounty palace, mar. 2001. wynei (2005). psathyrella spadiceogrisea (schaeff.) maire on mixed tree-litter, alevkaya, jan. 2001. wynei (2005). psilocybe crobula (fr.) singer on fallen pine-bark, st. hilarion, dec. 2000. on wood chips and plant remanins, rarely also on manure, autumn and winter. wynei (2005); loizides et al. (2011). uncommon. ramaria formosa (pers.) quél. in black pine forest, under golden oak (without date). loizides et al. (2011). rare. rectipilus cistophilus esteve-rav. & vila on a decaying twig of cistus creticus, agia paraskevi, 23 nov. 2011. torrejón (2013). rhizopogon luteolus fr. et nordholm under cistus spp. kormakiti forest, (without date). in light soils, (without site and date). in mixed forest especially under coniferous, alevkaya, autumn. nattrass (1937); wynei (2005); momany & gücel (2009). fairly common. rhizopogon roseolus (corda) th. [=rhizopogon vulgaris (vittad.) m. lange] on bare soil, karraagaç, nov. 1997. wynei (2005). annotated checklist of fungi in cyprus island 129 © the author(s) 2014 published by polish botanical society rhodocybe gemina (paulet) kuyper & noordel. in pinewoods, beside gecitköy lake and e of akdeniz, dec. 2001. wynei (2005). rhodocybe popinalis (fr.) singer on turf among pines, near ermeni evi below alevkaya, jan. 2001. wynei (2005). rigidoporus sanguinolentus (alb. & schwein.) donk on decaying wood of pinus nigra subsp. palliata, troodos, 17 nov. 2011. torrejón (2013). rigidoporus ulmarius (sowerby) imazeki [=fomes ulmarius (sowerby) fr.] on trunk of populus nigra. ay. nikolaos, paphos, 1933. at the trunk base of deciduous trees, usually carob, alevkaya, mar. nattrass (1937); momany & gücel (2009). as fomes ulmarius (sowerby) fr. in the first reference. ripartites tricholoma (alb. & schwein.) p. karst. on soil under pines, alevkaya, 1998. wynei (2005). occasional. rugosomyces carneus (bull.) bon [=calocybe carnea (bull.) kühner] amongst grass, alevkaya, winter. momany & gücel (2009). rugosomyces chrysenteron (bull.) bon under pinus sp., alrvkaya, dec. 1998. wynei (2005). rugosomyces onychinus (fr.) raithelh. in pinewoods, alevkaya, dec. 1998. wynei (2005). russula acrifolia romagn. in black pine forests, early in autumn. loizides et al. (2011). common. russula aurea pers. in black pine forests, early in autumn in high temperatures. loizides et al. (2011). frequent. russula chloroides (krombh.) bres. without data. in black and calabrian pine forests, as well as golden oak forests, early in autumn. loizides (2008); loizides et al. (2011). common. russula delica fr. without data. on pine-litter, yayla, nov. 2000. in all kinds of forests, early in autumn. wynei (2005); loizides et al. (2008, 2011). one of the commonest species in cyprus. russula insignis quél. associated with quercus and cistus, in early autumn. loizides & kyriakou (2011). russula luteotacta rea under oak an carob isolated among pines, antiphonitis, nov. 1997. in black and calabrian pine forests, as well as alluvial broadleaved forests, autumn. wynei (2005); loizides et al. (2011). common. russula medullata romagn. in black pine forests, very early in autumn in high temperatures. loizides et al. (2011). frequent. russula sanguinea (bull.) fr. in pinewood, tazik kiran and mt. yayla, dec. 1998. amongst grass in mixed forests with mostly coniferous trees, alevkaya, autumn. wynei (2005); momany & gücel (2009). russula torulosa bres. under pine (without site), dec. 2004. wynei (2005). 130 m. torrejón © the author(s) 2014 published by polish botanical society russula spp. without data. loizides (2008). sarcodon sp. under pine and near arbutus, opposite the picnic spot on the ridge road to kantara, dec. 2004. wynei (2005). schizophyllum commune fr. a common saprotroph on dead wood, (without date). on a willow stump, girne, (without date). on dead wood of deciduous trees and stumps, alevkaya, apr. nattrass (1937); wynei (2005); momany & gücel (2009). schizopora paradoxa (schrad.) donk on dead wood especially deciduous trees, alevkaya, apr. momany & gücel (2009). scleroderma verrucosum (bull.) pers. on soil beside a concrete step, karaman, nov. 1997. in heath, parks and woodland, alevkaya, autumn. in calabrian pine and alluvial forests, cultivated land and mediterranean maquis. wynei (2005); momany & gücel (2009). loizides et al. (2011). occasional. skeletocutis nivea (jungh.) jean keller on a dead pine branch, alevkaya, (without date). wynei (2005). common in the woods near alevkaya. sparassis crispa (wulfen) fr. on stumps and roots of various coniferous trees, autumn and winter. loizides et al. (2011). rare. stereum hirsutum (willd.) pers. a common saprotroph on dead wood, (without date). on olive trunk, (without site and date). on fallen branches and stumps of deciduous trees, selvilitepe, apr. nattrass (1937); wynei (2005); momany & gücel (2009). stereum ochraceoflavum (schwein.) sacc. on wood from decaying branches of quercus alnifolia, psilo dendra, 17 nov. 2011. torrejón (2013). stropharia aeruginosa (curtis) quél. found hidden among carobs, lapta, dec. 2000. wynei (2005). stropharia coronilla (bull.) quél. among reed-beds, özhan, dec. 1998. among lawns, parks and pasture, alevkaya, winter. wynei (2005); momany & gücel (2009). suillus luteus (l.) roussel in black and calabrian pine forests, autumn and winter. loizides et al. (2011). common. suillus bellinii (inzenga) watling in pinewoods under isolated carob and oak, esentepe, nov. 1999; jan. 2000. in black and calabrian pine forests, autumn and winter. wynei (2005); loizides et al. (2011). common. suillus bovinus (l.) roussel under coniferous trees, at the campus of the near east university, winter. momany & gücel (2009). annotated checklist of fungi in cyprus island 131 © the author(s) 2014 published by polish botanical society suillus collinitus (fr.) kuntze [=suillus fluryi huijsman] in mixed woodland forests, alevkaya, autumn. in black and calabrian pine forests, autumn and winter. momany & gücel (2009); loizides et al. (2011). common. as suillus fluryi huijsman in the first reference. suillus granulatus (l.) roussel on pine litter, alevkaya, jan. 2001. mycorrhizal in association with pine roots, at the campus of near east university; under pine trees, nicosia, dec. to jan. wynei (2005); momany & gücel (2009). suillus luteus (l.) roussel without data. loizides (2008). suillus mediterraneensis jacquet. & j. blum) redeuilh on pine litter, alevkaya, jan. 2001. wynei (2005). suillus tridentinus (bres.) singer under coniferous trees, alevkay, autumn. momany & gücel (2009). suillus spp. without data. loizides (2008). tapinella panuoides (batsch) e.-j. gilbert [=paxillus panuoides (fr.) fr.] on conifer stumps, karaman 2000; on ground debris probably attached to hidden roots, cengizköy 1998. on conifer woods stumps, viran kilise, (without date). wynei (2005); momany & gücel (2009). thanatephorus cucumeris (a. b. frank) donk [=corticium solani (prill. & declacr.) bourdot & galzin] on dianthus caryophyllus, (without date and site). zyngas( 1973 from georghiou & papadopoulos 1957). thanatephorus sterigmaticus (bourdot) p. h. b. talbot on a non decorticated dead stem of cistus creticus, foini, 21 nov. 2011. torrejón (2013). rare. thelephora caryophyllea (schaeff.) pers. under cistus, autumn, winter and spring. loizides & kyriakou (2011). tomentella asperula (p. karst.) höhn. & litsch. on a decaying acorn of quercus alnifolia, trail to calydonian falls, 19 nov. 2011. torrejón (2013). trametes pubescens (schumach.) pilát on an apple stump, lefke, feb. 1997. wynei (2005). trametes versicolor (l.) lloyd [=polystictus versicolor (l.) fr.], [=coriolus versicolor (l.) quél.] on dead wood of casuarina equisitifolia, nicosia, feb. 1935. nattrass (1937). on a carob stump, kozam, dec. 1998. on deciduous wood, alevkaya, during the hole year. wynei (2005); momany & gücel (2009). as coriolus versicolor (l.) quél. in the second reference. tremella mesenterica retz. on the cut surface of a pine, alevkaya, dec. 2001. wynei (2005). tricholoma albobrunneum (pers.) p. kumm. under pinus halepensis, troodos, feb. 1932. nattrass (1937). occasionally offered for sale in markets. 132 m. torrejón © the author(s) 2014 published by polish botanical society tricholoma caligatum (viv.) ricken without data. in black and calabrian pine, as well as golden oak forests, autumn and winter. loizides et al. (2008, 2011). frequent. tricholoma equestre (l.) p. kumm. in black and calabrian pine forests, autumn and winter. loizides et al. (2011). frequent. tricholoma fracticum (britzelm.) kreisel on pine litter, mt. yayla (without date). in calabrian pine forests, autumn and winter. wynei (2005); loizides et al. (2011). common. tricholoma portentosum (fr.) quél. deeply buried in pine litter, kozan, jan. 2001. wynei (2005). tricholoma spp. without data. loizides (2008). tricholoma terreum (schaeff.) p. kumm. in pinewoods, mid-winter, (whithout site). in black and calabrian pine forests, cedar forests, wood edges and mediterranean maquis, autumn and winter. under pine needles in mixed forests, karşiyaka, mar. wynei (2005); loizides et al. (2011); momany & gücel (2009). common. tubaria conspersa (pers.) fayod on soil and woody debris in a garden under oxalis pes-caprae, karaman nov. 1999. wynei (2005). tubaria dispersa (l.) singer [tubaria autochtona (berk & broome) sacc.] on soil, karaman, dec. 1998; under zizyphus bush, perhaps growing on its buried fruits, kanliköy. wynei (2005). tubaria furfuracea (pers.) gillet in manure grass among dittichia viscosa, özhan, dec. 1998; under zizyphus, kalavaç. wynei (2005). tulostoma squamosum pers. in turf around the ruined church, alevkaya, jan. 1999. wynei (2005). volvariella gloiocephala (dc.) boekhout & enderle in sheltered grassy place, especially if manure, alevkaya and karaman, dec. 1998. in fields, pastures and road edges. wynei (2005); loizides et al. (2011). frequent. vuilleminia comedens (nees) maire on a decaying branch of quercus alnifolia and on decaying branch of arbutus andrachne, psilo dendra, 17 nov. 2011. torrejón (2013). vuilleminia macrospora (bres.) horstam on wood from branches of cistus creticus, foini, 21 nov. 2011. torrejón (2013). vuilleminia megalospora bres. on a decaying branch of quercus alnifolia, trail to calydonian falls, 19 nov. 2011. torrejón (2013). rare. xerocomellus chrysenteron (bull.) šutara [=xerocomus chrysenteron (bull.) quél.] in black and calabrian pine forests, autumn. loizides et al. (2011). xerocomellus aff. chrysenteron [=xerocomus aff. chrysenteron] on the roadside pine litter, esentepe, nov. 2000. wynei (2005). xeromphalina cauticinalis (with.) kühner & maire [=xeromphalina fellea maire & malençon] under conifers, alevkaya, jan. 1999. wynei (2005). annotated checklist of fungi in cyprus island 133 © the author(s) 2014 published by polish botanical society conclusions at the time the author was looking for references and information dealing with fungi in cyprus island; the oldest documented information collected was chatin (1897). for this reason, it is supposed that the history of the mycology in cyprus started with this data. despite the fact that cyprus island is reported to be an area with rather scarce surveys and mycological literature, a good bunch of works dealing with fungi in this area is aported in this report. the most important contribution of the current paper is the compiled information broughting together different works dealing with fungi in this area throughout the three centuries of mycology in cyprus. several macrofungi labelled as rare in the notes of this checklist such as amanita torrendii, chlorophyllum molybdites, conferticium ochraceum, hyphoderma nemorale, myriostoma coliforme, rectipilus cistophilus, sparassis crispa, thanatephorus sterigmaticus, tomentella asperula and vuilleminia megalospora, should be considered under protection by local authorities. nevertheless, battarrea stevenii and boletus rhodoxanthus, wich are catalogued in many european countries as endangered species, appear to be very common in cyprus. nowadays, only a small group of local amateur mycologists are dealing with macrofungi in cyprus, and no one with microfungi. we are far away to know the mycobiota of the island, so it will be very important to organize new forays similar to british mycological association’s overseas meeting 2011 in order to improve the knowledge of fungi in this area. acknowledgements. the author would like to express his thanks to mr michael loizides, dr. salih gücel and dr. stephanos diamandis for providing cypriot bibliography. references altson r.a. 1956. report on an investigation into the cause of a root-rot of broad beens (vicia faba) in cyprus. department of agriculture. the government of cyprus, nicosia, cyprus. chatin m.a. 1897. un nouveau terfas (terfezia aphroditis) de l’ille de chypre. bulletin de la société botanique de france 44: 290. georghiou g.p., papadopoullos c. 1957. a second list of cyprus fungi. department of agriculture. the government of cyprus, nicosia, cyprus. loizides m. 2008. a secret world: the fungi of cyprus. field mycology 9 (3): 107-109. loizides m., kyriakou t. 2011. fungi of cistus maquis. field mycology 12 (1): 14-22. loizides m., kyriakou t., tziakouris a. 2011. edible and toxic fungi of cyprus. manitari, greece. loizides m., hobart c., konstandinides g., yiangou y. 2012. desert truffles: the mysterious jewels of antiquity. field mycology 13 (1): 17-21. momany a., gücel s. 2009. mushrooms of north cyprus, ecology, distribution, classification and toxicity. near east university, nicosia, north cyprus. momany a.t., mohamad a., gücel s. 2009. a comprehensive study on agaricus species of north cyprus. world journal of agricultural sciences 5 (2): 195-200. nattrass r.m. 1937. a first list of cyprus fungi. department of agriculture, the government of cyprus, nicosia, cyprus. nattrass r.m., papaioannou p. 1938. aditions to “a first list of cyprus fungi”. department of agriculture, the government of cyprus, nicosia, cyprus. neophytou g., ioannou n. 2009. first report of false smut disease on date palms in cyprus. journal of plant pathology 91 (1): 240. 134 m. torrejón © the author(s) 2014 published by polish botanical society torrejón m. 2013. fungi of cyprus: new data on microand macrofungi. acta mycol. 48 (2): 207-218. tsopelas p., nikolau k. 2005. first report of heterobasidion annosum in cyprus. plant pathol. 54 (4): 583. doi: 10.1111/j.1365-3059.2005.01202.x tsopelas p., angelopoulos a., nikolau k. 2008. seiridium cardinale is a new threat to cypress trees in cyprus. plant pathol. 57 (4): 784. doi: 10.1111/j.1365-3059.2007.01812.x willoughby l.g. 1983. the bacterial antagonist of karlingia rosea; further observations from spain and cyprus. nova hedwigia 38: 113-128. winey d.e. 2005. an illustrated introduction to the larger fungi of north cyprus. richmond publishing company, uk. zyngas j.p. 1973. the cyprus fungi. department of agriculture, ministry of agriculture and natural resources, nicosia, cyprus. 2014-06-30t23:05:57+0200 polish botanical society 2014-09-03t19:46:21+0200 polish botanical society 2014-09-03t19:47:51+0200 polish botanical society 2014-09-03t19:46:48+0200 polish botanical society 2014-08-31t21:56:30+0200 polish botanical society 2014-08-31t21:55:50+0200 polish botanical society 2014-09-05t16:23:27+0200 polish botanical society 2014-09-03t19:47:10+0200 polish botanical society 2014-09-06t20:25:17+0200 polish botanical society 2014-08-29t18:50:30+0200 polish botanical society 2014-09-05t16:23:47+0200 polish botanical society 2014-08-31t21:54:41+0200 polish botanical society note on the distribution of some lichenized and lichenicolous fungi of the tatra national park adam flakus laboratory of lichenology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl 31 512 kraków, ibflakus@ib pan.krakow.pl f l a k u s a .: note on the distribution of some lichenized and lichenicolous fungi of the tatra national park. acta mycol. 41(2): 329 342, 2006. new data about the occurrence of 25 species of rare lichens and 3 lichenicolous fungi in the tatra national park (western carpathians) are provided. of these species, fellhaneropsis vezdae is recorded for the first time from the whole tatra mts. and vezdaea stipitata is new to the polish tatra mts. the distribution of the species in the tatra national park is indicated. key words: lichens, lichenicolous fungi, biogeography, tatra mts., carpathians, poland introduction the tatra mts. are the greatest natural peculiarity of the west carpathians. these mountains are one of centre of biodiversity and occurrence of rare arcticalpine species in europe. at present, 1250 species of lichens and 87 species of allied fungi (lichenicolous fungi and saprotrophs) are known from the tatra mts. (western carpathians) (l i s i c k á 2005). however, the knowledge of the lichen biota of the once is still not complete and require more accurate research. especially, distribution of majority of species is in bad need of future attention. the present paper reports new information on the occurrence of 25 species of rare lichens and 3 lichenicolous fungi in the tatra national park. the currently known sites of the species in the area of the tatra national park are indicated in the maps (see figs 1–9). of presented species, fellhaneropsis vezdae is recorded for the first time from the whole tatra mts. and vezdaea stipitata is new to the polish tatra mts.. the results are based on material collected during research of the lichen biota in the tatra national park in the period of 2002–2005. herbarium specimens of the kram-l collection (kraków) were also taken into consideration. the specimens are deposited in the lichen herbarium of the w. szafer institute of botany, polish academy of sciences (kram-l) in kraków. the lichenicolous fungi are marked with an asterisk „*”. acta mycologica vol. 41 (2): 329-342 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 336 a. flakus leg. l. śliwa 3075; high tatra mts.: dolina za mnichem valley, alt. 1950 m, on granite stone, 21 august 2002, leg. a. flakus 104. normandina pulchella (borrer) nyl. in the polish tatra mts. the species has been recorded from dolina strążyska valley, on bryophytes over bark of fagus sylvatica, alt. ca 950 m by s u z a (1928) and fa ł t y n o w i c z (1999); dolina białego valley, alt. 960 m, on bark of acer pseudoplatanus by to b o l e w s k i (1956, 1957) and fa ł t y n o w i c z (1999); mt. łysanki, e slope above dolina strążyska valley, alt. ca 1050 m, on fagus sylvatica by to b o l e w s k i (1960); dolina olczyska valley, alt. 930 m, on fagus sylvatica by to b o l e w s k i (1962) and dolina za bramką valley, near mouth of a stream by fa ł t y n o w i c z (1999). the location of currently known sites of the species is indicated on the map (fig. 7). specimens examined. west tatra mts.: below mt. jastrzębia turnia near dolina nad capkami valley, alt. 1060 m, on bark of fagus sylvatica and corticolous mosses, 30 september 2004, leg. a. flakus 3545, on corticolous mosses on bark of acer pseudoplatanus, 30 september 2004, leg. a. flakus 3546. parmelina tiliacea (hoffm.) hale from the polish tatra mts. the species was reported by a l s t r u p and o l e c h (1992b), but the authors do not cite collecting sites (specimens not seen). the location of currently known sites of the species is indicated on the map (fig. 7). specimen examined. west tatra mts.: dolina chochołowska valley near siwa polana glade, alt. 925 m, on bark of acer pseudoplatanus, 3 may 2004, leg. a. flakus 2236. pertusaria flavicans lamy in the polish tatra mts. the species has been recorded from mt. skrajna turnia, alt. 1850 m, on vertical granite rock by a l s t r u p and o l e c h (1988). the location of currently known sites of the species is indicated on the map (fig. 7). fig. 7. distribution of ● normandina pulchella (borrer) nyl., ■ parmelina tiliacea (hoffm.) hale and ▲ pertusaria flavicans lamy in the tatra national park. 338 a. flakus contains calcium, 30 july 2005, leg. a. flakus 5125; dolina mułowa valley, lower part of mt. krzesanica, alt. 1900 m, on limestone rock, 9 july 2004, leg. a. flakus 2242. pseudosagedia guentheri (flot.) hafellner et kalb in the tatra national park the species has been recorded only from dolina rybiego potoku valley, w of czarny staw pod rysami, on overhanging granite, alt. 1610 m by a l s t r u p and o l e c h (1988). the location of currently known sites of the species is indicated on the map (fig. 1). specimen examined. high tatra mts.: dolina pięciu stawów polskich valley, by zadni staw lake, 49°12’39’’ n, 20°00’48’’ e, alt. 1895 m, on granite boulder among scree, in moist and shaded place, 18 august 2004, leg. a. flakus 3300. ramalina obtusata (arnold) bitter the species has been recorded from the polish tatra mts. by m o t y k a (1962), but the author do not mention stations, and by b i e l c z y k (2003) from dolina kościeliska valley, brama kantaka gate by potok kościeliski stream, on bark of picea abies, 6 july 1955, leg. j. nowak (kram-l 8266). the location of currently known sites of the species is indicated on the map (fig. 9). specimens examined. west tatra mts.: dolina kościeliska valley near the trail to wąwóz kraków gully, alt. 1090 m, on bark of picea abies, 22 july 2005, leg. a. flakus 5007, alt. 1100 m, on bark of picea abies, 22 july 2005, leg. a. flakus 5001. *thelocarpon epibolum nyl. in the tatra national park the species has been recorded from dolina chochołowska wyżnia valley, alt. ca 1180 m, mt. kobyła above dolina suchej wody valley, alt. ca 1180 m, brzeziny village by chowańcówka stream, alt. ca 1020 m, on rotten wood of picea abies by n o w a k (1974), mt. skupniów upłaz, alt. 1480 m, on aneura sp. on flat ground by a l s t r u p and o l e c h (1990) and siwa przełęcz pass, alt. 1815 m, on thallus of lichenomphalia hudsoniana by c y k o w s k a and f l a k u s (2005). the location of currently known sites of the species is indicated on the map (fig. 9). specimen examined. high tatra mts.: dwoisty żleb gully, 49°12’03’’ n, 20°04’44’’ e, alt. 1680 m, on thallus of baeomyces rufus, 6 august 2004, leg. a. flakus 2987/2. toninia athallina (hepp) timdal in the tatra national park the species has been recorded from n slope of mt. gładkie jaworzyńskie, on limestone by m o t y k a (1927). the location of currently known sites of the species is indicated on the map (fig. 2). specimen examined. west tatra mts.: dolina mułowa valley, lower part of mt. krzesanica, alt. 1900 m, on limestone rock, 9 july 2004, leg. a. flakus 2246. trapelia involuta (taylor) hertel distribution of the species in the polish tatra mts. has been characterized by f l a k u s (2004). the location of currently known sites of the species is indicated on the map (fig. 9). specimens examined. high tatra mts.: dolina pięciu stawów polskich valley, by zadni staw lake, 49°12’39’’ n, 20°00’48’’ e, alt. 1895 m, on small granite stones, 18 august 2004, leg. b. cykowska (kram-l 49750); sub-tatra trough: pańszczykowa polana glade near małe ciche village, 49°17’36’’ n, 20°03’36’’ e, alt. 920 m, on stones, 340 a. flakus b i e l c z y k u. 1997. materiały do flory porostów tatr ze zbiorów muzeum tatrzańskiego. fragm. flor. geobot. ser. polonica 4: 329 343. b i e l c z y k u . 1999. materiały do geograficznego rozmieszczenia porostów (lichenes) w polsce. 1. poro sty tatr. fragm. flor. geobot. ser. polonica 6: 245 253. b i e l c z y k u. 2003. the lichens and allied fungi of the polish western carpathians. (in:) u. b i e l c z y k (ed.). the lichens and allied fungi of the polish carpathians an annotated checklist: 23 232. c y k o w s k a b ., f l a k u s a . 2005. epigloea medioincrassata (epigloeaceae, non lichenized ascomycota), a species new to poland. polish bot. j. 50 (2): 233 234. c z a r n o t a p. 1998. some interesting lichens from gorce mts. (western beskidy mts.) new to poland. graphis scripta 9 (2): 59 61. c z a r n o t a p. 2002. caloplaca herbidella (hue) h. magn. (in:) u. b i e l c z y k , s. c i e ś l i ń s k i , w. f a ł t y n o w i c z (eds). atlas of the geographical distribution of lichens in poland. 3. w. szafer inst. bot., pol. acad. sci., kraków: 25 28. c z a r n o t a p. , k u k w a m . 2004. some sorediate lichens and lichenicolous fungi new to poland. gra phis scripta 15: 24 32. f a ł t y n o w i c z w. 1999. normandina pulchella (borrer) nyl. (in:) s. cieśliński, w. fałtynowicz (eds). atlas of the geographical distribution of lichens in poland. 2. w. szafer inst. bot., pol. acad. sci., kraków: 39 45. f a ł t y n o w i c z w. 2003. the lichens, lichenicolous and allied fungi of poland. an annotated checklist. w. szafer inst. bot., pol. acad. sci., kraków, 435 pp. f l a k u s a . 2004. new and rare lichen species of the polish tatra mountains. polish bot. j. 49 (1): 79 91. k i s z k a j . 2003. nowe dla pienin gatunki porostów. cz. iii. fragm. flor. geobot. polonica 10: 297 299. k u k w a m . 2004. porosty z rodzaju lepraria w tatrzańskim parku narodowym. parki nar. rez. przyr. 23: 3 12. l i s i c k á e . 2005. the lichens of the tatry mountains. veda, slovak academy of sciences, bratislava, 439 pp. m o t y k a j . 1926. die pflanzenassoziationen des tatra gebirges. vi teil: studien über epilitischen flechtengesellschaften. bull. int. acad. polon. sci., cl. sci. math., ser. b, sci. nat. 3 4: 189 227. m o t y k a j . 1927. materiały do flory porostów tatr. część ii. spraw. komis. fizyogr. akad. umiejętn. 61: 1 16. m o t y k a j . 1962. porosty (lichenes). 5 (2). (in:) flora polska. rośliny zarodnikowe polski i ziem ościennych. państwowe wydawnictwo naukowe, warszawa, 355 pp. n o w a k j . 1974. materiały do flory porostów tatr polskich. fragm. flor. geobot. 20 (1): 89 102. n o w a k j . , to b o l e w s k i z. 1975. porosty polskie. opisy i klucze do oznaczania porostów w polsce dotychczas stwierdzonych lub prawdopodobnych. państwowe wydawnictwo naukowe, warszawa kraków, 1177 pp. s u z a j. 1928. przyczynek do znajomości flory porostów polski. acta soc. bot. pol. 5 (2): 213 219. ś l i w a l. 2006. additions to the lichen flora of the tatra national park and its surroundings (polish carpathians). (in:) a. l a c k o v i č o v á , a . g u t t o v á , e. l i s i c k á , p. l i z o ň (eds). central eu ropean lichens diversity and threat. mycotaxon ltd, ithaca: 305 313. to b o l e w s k i z . 1956. lichenotheca polonica. fasc. vii. no 126 150. lichenes tatrenses. academia scientiarum poloniae, poznań, 9 pp. to b o l e w s k i z . 1957. materiały do flory porostów tatr ii. pozn. tow. przyj. nauk, wydz. mat. przyr. prace kom. biol. 17 (4): 1 22 + phot. 1 2. to b o l e w s k i z . 1960. materiały do flory porostów tatr iv. pozn. tow. przyj. nauk, wydz. mat. przyr. prace kom. biol. 21 (5): 1 31. to b o l e w s k i z . 1962. materiały do flory porostów tatr v. pozn. tow. przyj. nauk, wydz. mat. przyr. prace kom. biol. 24 (2): 21 30. wę g r z y n m . 2006. porosty apofityczne (apoporosty) w dolinie suchej kasprowej w tatrzańskim parku narodowym. parki nar. rez. przyr., 25 (2): 3 10. note on the distribution of some lichenized fungi 341 materiały do rozmieszczenia porostów i grzybów naporostowych tatrzańskiego parku narodowego s t r e s z c z e n i e tatry stanową największą osobliwość przyrodniczą karpat zachodnich. są jednym z cen trów różnorodności gatunkowej, jak również miejscem występowania rzadkich arktyczno al pejskich gatunków w europie. obecnie znanych jest z tego pasma 1250 gatunków porostów i 87 gatunków związanych z nimi grzybów naporostowych oraz saprobiontów (l i s i c k á 2005). mimo to, wiedza dotycząca lichenobioty tatr jest wciąż niekompletna i wymaga dalszych syste matycznych badań. w pracy przedstawiono nowe stanowiska 25 rzadkich gatunków porostów i 3 grzybów naporostowych z tatrzańskiego parku narodowego wraz z ich rozmieszczeniem na tym obszarze (figs 1 9). fellhaneropsis vezdae jest gatunkiem nowym dla polskiej części karpat i jednocześnie dla całego pasma tatr, a vezdaea stipitata dla tatr polskich. 2014-01-01t11:44:56+0100 polish botanical society 2014-09-02t20:11:03+0200 polish botanical society 2014-09-05t16:23:02+0200 polish botanical society 2014-09-06t20:25:13+0200 polish botanical society 2014-09-08t11:11:46+0200 polish botanical society 2014-08-31t21:56:53+0200 polish botanical society 2014-09-05t16:24:35+0200 polish botanical society 2014-08-31t21:55:21+0200 polish botanical society 2014-09-02t20:09:25+0200 polish botanical society 2014-08-31t21:57:23+0200 polish botanical society 2014-08-31t21:53:28+0200 polish botanical society 2014-09-03t19:47:41+0200 polish botanical society 2014-09-02t20:10:20+0200 polish botanical society 2014-08-31t21:55:01+0200 polish botanical society the development of erysiphe alphitoides and e. hypophylla in the urban environment ewa sucharzewska department of mycology, university of warmia and mazury in olsztyn oczapowskiego 1a, pl-10-719 olsztyn-kortowo, ewko@uwm.edu.pl sucharzewska e.: the development of erysiphe alphitoides and e. hypophylla in the urban environment. acta mycol. 44 (1): 109–123, 2009. differentiated responses of erysiphe alphitoides and e. hypophylla in urban conditions are described. the influence of transport pollution on the morphology of the mycelium, chasmotecium development and individual stages of the developmental cycle is discussed. key words: erysiphales, developmental cycle, transport pollution, olsztyn introduction erysiphe alphitoides griff et maubl. and e. hypophylla nevod. are common obligate parasites of the order erysiphales. they infect representatives of the genera quercus and fagus as well as sporadically aesculus and castanea. paeonia lutea has been identified as a new host of erysiphe hypophylla (takamatsu et al. 2006). the two phytopathogens are widespread in entire europe, asia, america, australia and new zealand. the occurrence of erysiphe alphitoides has been documented in poland since 1909, initially in the conidial stage only (oidium quercinum thüm). the sexual stage of e. alphitoides was first observed in france in 1911 and recorded in poland in 1922 (braun 1995; sałata 1985). it has since been recorded in the teleomorphic stage as microsphaera alphitoides very often (adamska et al. 1999; czerniawska 2001; dynowska et al. 1999; kalinowska‑kucharska, kadłubowska 1993; kućmierz 1967, 1971, 1973; majewski 1970, 1971; mułenko 1981, 1988; mułenko, wojdyło 2002; ruszkiewicz‑michalska 2006; sałata et al. 1993). erysiphe alphitoides is a dangerous oak pathogen. it attacks trees of various age classes causing a reduction in annual growth (minkievič et al. 1993). acta mycologica vol. 44 (1): 109–123 2009 110 e. sucharzewska erysiphe hypophylla was first observed in poland in 1952 (sałata 1985). it has been recorded sporadically in different parts of poland (dynowska et al. 1999; ma‑ jewski 1971; mułenko 1988; sałata, majewski 1976). in contrast to erysiphe alphitoides, the systematic position of e. hypophylla was not clear until recently (braun, takamatsu 2000). the species was considered by many mycologists to be a synonym of erysiphe alphitoides (microsphaera alphitoides) based on biometric studies. braun et al. (2003) finally distinguished two separate species using molecular studies: erysiphe alphitoides and erysiphe hypophylla. the two fungi, mostly erysiphe alphitoides and less frequently e. hypophylla, has been reported in studies investigating their presence in floristically or phytosocio‑ logically interesting communities and ecosystems characterised by a low level of an‑ thropopressure. as they were often inventories or phytopathological examinations, these studies usually provided systematic and geographical analyses of species rich‑ ness or an assessment of taxonomic differentiation in relation to plant associations in a given area (adamska et al. 1999; czerniawska 2001; majewski 1971; mułenko 1996, 1998; kućmierz 1971, 1973). however, the ecology erysiphe alphitoides and e. hypophylla and their life strategies in environments exposed to strong anthropopres‑ sure have been discussed in few studies. the aim of this study was to analyse the occurrence of erysiphe alphitoides and e. hypophylla and to examine basic life reactions of the parasites in urban conditions. material and methods examinations (2000‑2002) were conducted in the city of olsztyn and its vicinity at 63 sites (17 sites with young plants and 46 sites with mature quercus robur l. plants) situated at a range of distances from main transport routes: up to 50 m, up to 100 m, up to 300 m and >300 m. distances were selected using studies by lorenc‑plucińska and byczyńska (1997) which show that the greatest value of exhaust gas concentra‑ tion is recorded at a distance of 30‑50 m away from the road and is at a constant level of 30%. the exhaust gas level drops to 10% at 200 m away from transport routes. sites located >300 m were used as controls. material was obtained from the middle of the vegetative period when first chas‑ mothecia began to occur on the powdery mycelium. one sample was defined as a total of 25 leaves collected randomly from each host plant. the infection rate of host plants, mycelium development and morphology as well as chasmothecium development were examined in the macro‑ and microscopic analysis of the material. 1. the disease rating was calculated for each sample on a 5°‑scale using mckin‑ ney’s formula. it shows the degree of infection of host plants and is a criterion of pathogen sensitivity (dynowska 1993, 1994) to changes in the optimum environment conditions: σ (a x b) x 100% r = ------------------- n x 4 erysiphe alphitoides and e. hypophylla 111 r – disease coefficient in percent (index); σ (a x b) x 100% – the sum of the products obtained by multiplying the number of plant organs (a) by the degree of infection (b); n – total number of plants (alternatively leaves, fruits) examined in the study; 4 – the highest degree of infection in a five‑grade scale (0 – no infection; 1 – up to 10%; 2‑11 – 25%; 3‑26 – 50%; 4‑51 – 100%). the final value r used in the results analysis was calculated based on the arithmetic mean for each fungal species on a specific host plant = mean degree of infection. 2. the developmental stage of the parasite was identified: asexual stage, sexual stage. 3. the number of chasmothecia, both mature and immature, per 1 cm2 of the sur‑ face of each infected leaf was established. ten randomly collected morphologically mature chasmothecia were selected and analysed under a microscope. the following were assessed: a) developmental degree of appendages in a three‑grade scale: 0 – chasmothecia without appendages, i – chasmothecia with appendages not fully developed, ii – chasmothecia with fully developed appendages; b) maturity of chasmothecia in a three‑grade scale: 0 – chasmothecia without asci and spores; i – chasmothecia with asci without formed ascospores; ii – chasmothecia with asci containing spores; c) morphological variability of chasmothecia, the size and diameter of chasmoth‑ ecia, appendages. the number of chasmothecia discussed in the results was calculated based on the arithmetic mean for each fugal species. fungi were determined using keys by braun (1987, 1995) and sałata (1985). spe‑ cies were identified using morphological traits of chasmothecia (size and diameter, number of appendages, appendages branching). the nomenclature was accepted after braun and takamatsu (2000) and braun et al. (2003). host plants were determined using studies by szafer et al. (1988) and rutkowski (1998). statistical calculations were conducted using the data analysis software system statistica (statsoft, inc.) version 6 (2003) by variance analysis. significant differ‑ ences among the means were declared at a significance level of p=0.05. the means were then classified in homogenous groups for selected factors using duncan’s test. letter symbols (a, b, c…) were used and values to which the same letters were as‑ signed do not differ significantly at p=0.05. results erysiphe alphitoides and e. hypophylla were observed on both mature and young quercus robur plants in all the years of the study period: at 40 sites in 2000 (63%), at 47 sites in 2001 (75%) and at 48 sites in 2002 (77%) (fig.1). the analysis of the degree of infection of the host plant in relation to the distance from transport routes shows noticeable, statistically significant differences. the high‑ est mean disease index was recorded at sites located up to 100 m and 50 m (52% and 112 e. sucharzewska 44%, respectively). the lowest mean infection was observed at sites located up to 300 m and >300 m, which is confirmed by the statistical analysis (fig. 2). the two parasites either occurred separately or co‑occurred on the same plant and even on the same leaf. the upper leaf surface was usually colonised by e. alphitoides and the lower leaf surface by e. hypophylla. chasmothecia of e. alphitoides were recorded beside chasmothecia of e. hypophylla on the lower leaf surface in a few cases. a relatively high participation of the species’ co‑occurrence on the same leaves was recorded in each study year. changes in the prevalence of the two fungi were also observed. e. alphitoides increased its occurrence at the distances examined in the study year by year. the highest participation of the species was recorded at sites located up to 50 m. e. hypophylla occurred more frequently at sites located up to 300m and >300m (fig. 4). two developmental stages of e. alphitoides and e. hypophylla were observed throughout the study period: the asexual stage when conidial spores occurred and the sexual stage when chasmothecia were produced. samples containing the perfect stage of the two fungi constituted the highest percentage regardless of the distance (>70%). fig. 1. percentage participation of e. alphitoides and e. hypophylla in samples on q. robur. fig. 2. mean degree of infection of q. robur by e. alphitoides and e. hypophylla in relation to the distance from transport routes in the study period (letters a and b indicate sta‑ tistically significantly different val‑ ues at p=0.05). fig. 3. mean number of e. alphitoides chasmothecia per 1cm2 of leaf surface throughout the study period. 118 e. sucharzewska e. alphitoides in areas strongly influenced by pollution has also been observed by boczoń (1998) and domański et al. (1987). divergent results of research into the occurrence of e. alphitoides is explained by domański (1976). he attributes epiphy‑ toses to a specific system of ecological factors that favourably influence the devel‑ opment as well as aggressiveness and pathogenecity of the disease agent. a set of factors co‑interacting in different combinations most probably occurred in each of the above cases and may have influenced divergent study results. this was shown in a study by grzebyta et al. (2005), who analysed the influence of high temperature and fluoride on the development of e. alphitodes on q. robur: higher infection of leaves in oaks growing in an area containing fluoride than in a clean area was recorded in previous observations. this indicated a low sensitivity of the pathogen to pollution with fluoride compounds. more detailed examinations, however, showed that fluo‑ ride which acted synergistically with high temperature caused the opposite effect and led to low infection of leaves. erysiphe hypophylla, which mostly colonised the lower leaf surface similarly to e. alphitoides, developed in all the distance zones. its strongest development was observed at sites up to 300 m and at control sites. interestingly, the species co‑oc‑ curred on the same leaves and the percentage of the occurrence of this type was high in the distance zones in the three study years. information on the occurrence of biology of e. hypophylla is scarce in the available literature. it was recorded in the białowieża national park by majewski (1971) and at a few localities in poland by sałata & majewski (1976) and dynowska et al. (1999). the species seems to be common and the present study shows that it often occurs along with e. alphitoides. however, the presence of the latter is recorded in many polish studies (adamska et al. 1999; czerniawska 2001; kalinowska‑kucharska, kadłubowska 1993; mułenko, wojdyło 2002). these studies are conducted mostly in environments similar to natu‑ ral ones where the degree of anthropogenecity is low. these results, however, cannot be used to define the occurrence of e. hypophylla in urbanised and natural areas. di‑ vergent opinions on the separate position of the two parasites and their treatment as one species, erysiphe (microsphaera) alphitoides, may have also contributed to this. weather conditions also influenced the development of the two species in the study period. erysiphales can infect plants in a broad range of temperatures and humidity. due to a considerable amount of water in the cell, spore germination in the majority of the representatives of the order takes places even in low relative air humidity. the highest degree of infection in all the species analysed in the study was observed in 2002 when the vegetative period was very warm and dry. july, au‑ gust and september were hot while precipitation between may and september was lower than usual. similar results were obtained by durska (1974), who recorded an increased development of erysiphales during a dry and hot summer. parasites aim to close the developmental cycle in order to produce structures having the best possible resistance genes that condition the survival of a species. therefore, a strategy aimed at increasing and improving reproduction is a char‑ acteristic trait of parasites. organisms whose features are better adapted to their environments achieve a greater reproduction success. the sexual process may be disturbed in unfavourable conditions, such as pollution, which may result in the ab‑ sence of or a reduction in the number of ascomata or degeneration. this was ob‑ served in lophodermium pinastrii by grzywacz (1976) as well as by kowalski and erysiphe alphitoides and e. hypophylla 119 budnik (1976). the parasite could not reach a full developmental cycle due to high concentrations of toxic compounds and altered chemism of infected needles, and did not produce ascomata. a full developmental cycle of the two powdery mildews that ended in the produc‑ tion of chasmothecia, also called cleistothecia, characteristic of this fungal group, was observed in the present study. chasmothecia in the order erysiphales are scle‑ rotia and their initiation occurs in unfavourable environmental conditions accord‑ ing to some researchers (dynowska 1996a; füzi 2001). literature data show that the majority of powdery mildews enter the reproduction stage towards the end of the vegetative period, between august and november (füzi 2001; majewski 1971; mmbaga 2002; sałata 1985; turnau, czerwonka 1986). present results corresponds with literature reports. young chasmothecia in erysiphe alphitoides and erysiphe hypophylla were observed at most sites at the beginning of august. they began to ma‑ ture in the second half of august and mature chasmothecia were recorded in mid september. similar results were obtained by kadłubowska, kalinowska‑kucharska (1989) and minkievič et al. (1993). however, the parasites did not always enter the generative stage. the fact that the oidium form was recorded towards the end of the study periods suggests that the sexual stage is not obligatory in the species and the parasite also overwinters in the anamorphic form. majewski (1971) also ob‑ served that not all powdery mildews enter the teleomorphic stage in his studies in the białowieża national park. statistically significant differences in the number of chasmothecia between the zone influenced by automotive exhaust gases and the control zone were observed in erysiphe alphitoides. parasites formed chasmothecia more numerously at a distance up to 50 m away from transport routes and their smallest number was recorded >300 m. this may result from the differences in the degree of infection of the host plant. a similar opinion was expressed by füzi (2001), who observed a strict correlation between the number of chasmothecia and the degree of infection: both variables were mutually dependent. mmbaga (2002) disagrees and reports that neither the degree of infection of the host (leaves with a low infection degree often contained a high number of chasmothecia) nor the plant age influences the number of cha‑ smothecia. significant disturbances in chasmothecium development at sites located by main transport routes were not observed in the two powdery mildews. it is interesting that differences in appendage development were observed be‑ tween the two mildews infecting q. robur. a considerably greater number of chas‑ mothecia without appendages (0 developmental stage) and having appendages not fully developed (i developmental stage) were recorded in e. alphitoides. chasmo‑ thecia having appendages in i and ii developmental stage dominated in e. hypophylla. the lower leaf surface may have been more favourable for the development of chas‑ mothecia. this is reflected in the maturity of asci and spores. a high participation of chasmothecia without developed asci and spores was recorded in e. alphitoides while chasmothecia with asci containing spores constituted a considerably higher percentage in e. hypophylla. untypical chasmothecia whose size was considerably smaller and which did not have developed asci and spores but had fully developed appendages and chasmothecia with untypical appendages were observed in erysiphe alphitoides. these chasmoth‑ ecia were recorded in the zone up to 50 m in 2002. empty chasmothecia have been 120 e. sucharzewska described in epichlöe typhina in athropogenic populations of pucciniella distans and attributed to disturbances in the genetic control of the developmental cycle of e. typhina caused by an environmental factor (falińska 2002). degenerated chasmothecia of a considerably smaller size were observed in lophodermium pinastrii in the zone of the strongest influence of pollution by benben, sierota (1976) and grzywacz (1976). empty chasmothecia as well as untypical appendages in polluted sites recorded in the present study may indicate developmental disturbances. however, such chasmoth‑ ecia constituted a very low percentage. their occurrence may thus be caused natural disturbances resulting from the formation of a great number of chasmothecia or the influence of higher temperatures as suggested by moore‑landecker (1992). ecological research into the parasites of the order erysiphales conducted in the urban environment does not unambiguously show the influence of individual factors on the development of the fungi examined in this study. however, the two species have a broad ecological amplitude. this is connected with adaptation processes con‑ ditioned by genetic mechanisms that are activated at times of greater and more long‑ term amplitudes of environmental factors that are characteristic of urbinocenoses (stearns 1992). conclusion the absence of major disturbances in the life cycles of e. alphitoides and e. hypophylla, the occurrence of all developmental stages with the predominance of the 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(strain td50) has been tested in vivo against the main phytopathogens of rape: sclerotinia sclerotiorum (lib.) de bary, botrytis cinerea pers., alternaria spp. and fusarium spp. in greenhouse at the laboratory of mycology and plant pathology, biology faculty, university of bucharest – romania and in the field at the agricultural experimental research-development station caracal (aerds), olt district. the t. viride (strain td50) bioproduct formulated as a powder for the seed treatment has been effective in the protection of rape plantlets against the above mentioned phytopathogens. key words: rape, seedand soil-borne fungi, antagonistic fungi, biocontrol, trichoderma, phytopathogens, sclerotinia sclerotiorum, botrytis cinerea, alternaria spp., fusarium spp., bioproduct from t. viride (strain td50), romania introduction in the frame of integrated protection of rape crop, aspects concerning chemical control of pathogens, pests and weeds have been published in romania by different authors (baicu, săvescu 1986; iliescu et al. 1987; oancea 1998; bîlteanu 2001; diaconu, mateiaş 2004; hălmăjan 2006; mantu 2007; popov, raranciuc 2007). also, some steps for diversification of the protection means have been recorded, among them the biological non-polluting ones using antagonistic fungi (şesan 1992, 2001, 2005; şesan, groza 2007; şesan 2008). the objective of this approach was the evaluation of potential antagonistic ability of some trichoderma viride isolates against the main pathogens of rape crop: sclerotinia sclerotiorum (lib.) de bary, botrytis cinerea pers., alternaria spp., fusarium spp. acta mycologica vol. 49 (1): 87–92 2014 doi: 10.5586/am.2014.008 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 88 t.-e. şesan © the author(s) 2014 published by polish botanical society material and methods the investigations for evaluation of potential antagonistic ability of some t. viride isolates against the main rape phytopathogens have been conducted in vitro and in vivo (greenhouse, experimental field). in vitro method of double cultures on the same medium in petri plates it was used, after jouan et al. (1964), presented in the figure 1 the antagonistic fungal ability has been expressed by the coefficient x, calculated after the formula x = ia/ib x eb/ea. in this formula the coefficient x is the value of the quotient of inner radius (i) and outer radius (e) of the test-fungus (a) and the antagonistic fungus (b). in case of x = 1, no influence has been expressed between the two tested fungi; when x < 1, the antagonism is stronger when the coefficient x value is lower or close to 0 (zero); when x > 1, the tested isolates prove no antagonism against the checked phytopathogens. for in vitro experiments, fungal isolates of sclerotinia sclerotiorum, botrytis cinerea, fusarium spp., alternaria spp. and trichoderma spp. from rape seeds have been used. as antagonistic fungi, 5 strains of trichoderma viride (td5, td35, td45,td49, td50), isolated by the author, have been tested. the bioproduct from trichoderma viride isolatetd50 (şesan, oancea 2010; şesan et al. 2012) has been also, tested in greenhouse at the university of bucharest, laboratory of mycology and plant pathology for the treatment of rape seeds in comparaison with an untreated control and with a chemical standard – procymidone 50. the experiment has been performed in sterile soil, in petri plates of 12 cm diameter, each variant having 5 replicates. a number of 10 rape seeds hydromel cv. (from aerds caracal, olt district, 2007) have been used for each replicate. the application dose for biological seed treatment was 1g powder of t. viride bioproduct per kg seeds. in the field at aerds caracal, olt district, the experimental variants were as follows: 1) seeds treated with trichosemin 25 pts – 1 g/kg seed; 2) seeds treated with a chemical standard (procymidone 50) – 1 g/kg seed and 3) untreated control. the manitoba cv. was cultivated. fig. 1. method of evaluation of the relationships between potential antagonistic fungi and fungal phytopathogens (jouan et al. 1964). mycobiota of rape seeds in romania. ii. 89 © the author(s) 2014 published by polish botanical society in this experiment, evaluation of number and percent of emerging healthy plantlets in comparaison with the same parameter in the standard variant (chemical fungicide procymidone 50) and in the untreated control was performed. results and discussion in vitro. among the tested isolates (tab. 1), the strain td35 proved the strongest antagonism against tested phytopathogens, the value of x coefficient ranging between 0.22 and 0.44 (fig. 2). table 1 evaluation of the antagonistic activity of trichoderma viride strains based on x coefficient (after jouan et al. 1964) trichoderma viride fusarium sp. alternaria alternata botrytis cinerea sclerotinia sclerotiorum td35 0.25 0.40 0.44 0.22 td45 0.78 0.86 0.70 0.48 td49 0.28 0.90 0.62 0.76 td50 0.30 0.42 0.35 0.54 td5 (control) 0.55 0.54 0.89 0.45 in the decreasing order, a good antagonistic ability have been proved for strains td50 (x = 0.30-0.70) and td49 (x = 0.30-0.89). the lowest antagonistic activity has been noticed for the isolates td45 , with x coefficient between 0.48 and 0.86 and td5 (control), with x coefficient between 0.45-0. 89. the antagonistic behaviour of the tested isolates is shown by the following decreasing order: td35>td49>td50>td45>td5. in vivo. the experiments performed under greenhouse conditions (tab. 2, fig. 3) proved that the percent of emerging healthy plantlets, in comparison with the untreated control, was higher by 47%. fig. 2. double cultures obtained by jouan et al. (1964) method, with phytopathogens botrytis cinerea (a), fusarium sp. (b), sclerotinia sclerotiorum (c), as test-fungi and trichoderma viride, strain td50, as antagonistic fungus. a b c 90 t.-e. şesan © the author(s) 2014 published by polish botanical society table 2 testing the trichoderma viride (td50) bioproduct for protecting rape crop cv. hydromel against soiland seed-borne pathogenic fungi in greenhouse (2007 yield) variant emergence of healthy plants after 8 days difference in comparaison with control (%) no. seedlings % seedlings %seedlings 1. t. viride (td50) bioproduct – 1 g/kg seed 100 147 +47 2. chemical standard (procymidone 50) – 1g/kg seed 90 130 +30 3. control (untreated) 70 100 in the standard variant (procymidone 50), the value of the emerging and healthy rape seedlings was higher by 30% in comparaison with the untreated control. under the field conditions, in the variant with biological seed treatment, the percentage of emerging helthy plantlets was higher by 26% in comparaison with the untreated control and similar to the variant of the chemical standard (tab. 3). however, after 14 days (29th august 2008), no differences have been registered between the biological and chemical treatments. these results obtained in the rape crop were similar to other experimental results obtained by us for other industrial oilcrops (sunflower, soybean) and annual pulses (bean, soybean, cickpea) (baicu, săvescu 1986; şesan et al. 1997 a, b) and to our previous results in the rape crop (galani et al. 2008). also, these results confirm our tests in vivo proving the efficacy of our bioproduct with t. viride (td50), patented in 2012 (şesan et al. 2012) for protecting oilseed plants among them rape (şesan, oancea 2010). in order to obtain a healthy, non-polluted and productive rape crop, the instructions for the application of biological control of rape seeds with the bioproduct based on t. viride were prepared. these instructions consist of: 1) seed treatment: dry; 2) dose of treatment: 1-2 g/kg seeds; 3) time of treatment application: 1-2 days before sowing; 4) storage conditions for the bioproduct: in the dry, wellaired spaces, with a good ventilation, at low temperatures, in the shadow, avoiding untreated control bioproduct from trichoderma viride (td50) 1 g/kg seed chemical standard (procymidone 50) 1 g/kg seed fig. 3. testing the protective activity trichoderma viride bioproduct (td50). mycobiota of rape seeds in romania. ii. 91 © the author(s) 2014 published by polish botanical society the direct; 5) proper conditions for bioproduct transport, protected against high temperatures and humidity. these instructions are very important for the agricultural practice. conclusions 1. the antagonistic ability of 5 trichoderma viride strains (td5, td35, td45,td49, td50) against 4 species of phytopathogens isolated from rape seeds (fusarium sp., botrytis cinerea, alternaria alternata, sclerotinia sclerotiorum) was evaluated in vitro. the antagonistic ability of strains was evaluated in decreasing order as: td35 >td49 > td50 > td45 > td5. 2. bioproduct with trichoderma viride – td50 strain – was efficient in the rape protection against the seedand soil-borne pathogens (sclerotinia sclerotiorum, botrytis cinerea, fusarium spp., alternaria spp.) in the greenhouse (hydromel cv.) and in the field (manitoba cv.), too. applied as seed treatment at a rate of 1-2 g/kg seeds, t. viride bioproduct stimulated the emergence of plantlets and their health status. 3. the instructions for the application of biological control of rape seeds with the bioproduct based on t. viride were proposed: 1) seed treatment: dry; 2) dose of treatment: 1-2 g/kg seeds; 3) time of treatment application: 1-2 days before sowing; 4) storage conditions for the bioproduct: in the dry, well-aired spaces, at low temperatures, in the shadow, avoiding the direct; 5) proper conditions for bioproduct transport, protected against high temperatures and humidity. these instructions are very important for the agricultural practice for obtaining a healthy, productive, nonpolluted with chemicals rape crop. acknoledgments. the author is very grateful for the financial support of this research, in the frame of the project ceex 75 agral (2006-2008), coordinated by the national council of scientific research in romania. also, she thanks to the senior scientist dr. n. vilău, from the aerds caracal – olt district, for the field test presented in table 3. table 3 testing trichoderma viride (td50) bioproduct for protecting rape plants (manitoba cv.) against seedand soil-borne pathogens under the field conditions aerds caracal – olt district 2008 variant emergence of healthy plants after 8 days difference to the control (%) no. plantlets % no. % 1. t. viride bioproduct – 2 g/kg seed 86 126 +18 +26 2. procymidone 50 (chemical standard) – 2 g/kg seed 88 129 + 20 + 29 3. untreated (control) 68 100 92 t.-e. şesan © the author(s) 2014 published by polish botanical society references baicu t., săvescu a. 1986. systems of integrated control against crop diseases and pests [in romanian], ed. ceres, buc.: 264 pp. bîlteanu gh. 2001. phytotechny [in romanian]. vol. 2, ed. ceres, buc.: 90-108. diaconu p., mateiaş m.c. 2004. agricultural technologies – rape and mustard crops [in romanian], ed. geea, buc., 25 pp., 27 color figs. galani g., şesan t.-e., trotuş e., vilău n. 2008. elements of biological protection of rape crop [in romanian]. annual scientific session of the research-development institute for plant protection bucharest, romania "methods of integrated protection of agricultural crops – important factor for the food security and safety", academy of agricultural sciences and forestry, 29th may 2008, programme and book of abstracts: 21-22. hălmăjan v. (ed.). 2006. the guide of the rape producer [in romanian], ed. agris, red. rev. agr. buc.: 203 pp. iliescu h., iordache emilia, ioniţă alina, jinga v., 1987, main rape diseases and their control [in romanian]. buc., ma, asas, ccpp, booklet: 10 pp. jouan b., lemaire j.m., arnoux j. 1964. éléments d’appréciation des intéractions entre champignons cultivés in vitro, phytiatrie-phytopharmacie 13: 185-195. mantu i. 2007. new sunflower and rape genotypes registered by istis in 2007 [in romanian]. sănătatea plantelor 110 (7): 7. oancea i. 1998. treatise of agricultural technologies [in romanian, ed. ceres, buc.: 182-185. popov c., raranciuc s. 2007. protection of autumn rape crop [in romanian]. sănătatea plantelor 111 (8): 8. şesan t. 1992. bibliography of romanian contributions in the field of biological control of plant mycoses [in romanian]. probl. prot. plant. xx(1-2): 85-96. şesan t.-e. 2000/ publ. 2001. biological control, advantages, disadvantages, integration [in romanian]. symposium dedicated to the academician gh. ionescu-şişeşti on the occasion of his 115 years since he was born, 26.10.2000, asas, volume "priorities of the scientifical researches in the field crops" [in romanian], ed. ceres, buc.: 143-150. şesan t.-.e. 2005. romanian bibliography in the field of biological control of plant mycoses [in romanian]. sănătatea plantelor, special edition – august 2005: 15-22. şesan t.-e. 2008. evaluation of fungal diversity of rape seeds in the south of romania, book of abstracts, mp-171, the xiith international congress of mycology iums turkey, istanbul, 5-9th august 2008: 36 . şesan t.-e., csép n., oancea f., procopovici e., raranciuc s., ivancia v. 1997a. biological protection of sunflower against white ro (sclertotinia sclerotiorum) [in romanian, probleme de protecţia plantelor, xxv (1): 27-44. şesan t.-e., csép n., oancea f., procopovici e., guran m. 1997b. biological protection means of annual pulses against seedand soil-borne pathogens [in romanian], probleme de protecţia plantelor, xxv (2): 245-256. şesan t.-e., groza o. 2007. mycobiota associated with rape seeds in romania. abstracts of the xvth congress of the european mycological association (ema), 16-22nd september 2007, saint petersburg (russia): 264. şesan t.-e., oancea f. 2010. trichoderma viride pers. – experimental model for biological and biotechnological investigations of mycromyceta with importance in obtaining plant protection bioproducts. journal of plant development 17: 49-62. şesan t.-e., oancea f., ştefan a.-l., lupu c., iliescu h. 2012. antagonistic trichoderma viride strain and the procedure for obtaining an antifungic bioproduct based on this [in romanian], patent nr. 126125/ 28.12.2012: 6 pp. 2014-06-30t22:38:45+0200 polish botanical society three lichen species of micarea (pilocarpaceae) new to belarus 249this is an open access article distributed under the terms of the creative commons attribution 3.0 license (creativecommons.org/licenses/by/3.0/), which permits redistribution, commercial and non-commercial, provided that the article is properly cited. © the author(s) 2014 published by polish botanical society acta mycologica introduction the genus micarea was described by fries [1] to accommodate a crustose species with a green granular thallus and convex immarginate apothecia named m. prasina fr. currently the genus comprises ca. 100 species [2,3] and is considered to be polyphyletic combining a large number of phenotypically variable species, as well as a heterogeneity of infrageneric characters [4,5]. further studies will probably provide stronger support for the delimitation of at least some distinct phylogenetic lineages obtained by andersen and ekman [5] and based on mtssu sequences, will legitimate descriptions of several new genera. some steps towards a new nomenclature of these species have recently started, with harris [6] transferring micarea erratica (körb.) hertel, rambold & pietschm. into the new genus leimonis r.c. harris, and ekman and svensson [7] introducing the genus brianaria s. ekman & m. svensson for species of the former micarea sylvicola group. nearly 60 species of micarea s.lat. are currently known from europe (e.g. [2,8–13]), of which only 10 have been reported from belarus, namely m. cinerea (schaer.) hedl., original research paper acta mycol 49(2):249–253 doi: 10.5586/am.2014.018 received: 2014-05-24 accepted: 2014-10-03 published electronically: 2014-12-02 three lichen species of micarea (pilocarpaceae) new to belarus andrei tsurykau1*, paweł czarnota2 1 department of biology, f. skorina gomel state university, sovetskaja 104, 246019 gomel, belarus 2 department of agroecology, university of rzeszów, ćwiklińskiej 2, 35-601 rzeszów, poland abstract micarea elachista, m. micrococca and m. misella are reported for the first time from belarus. their phenotypic characters, distribution and ecological preferences are given. keywords: ascomycota; lichenized fungi; crustose lichens; biodiversity; pine forest; gomel region * corresponding author. email: tsurykau@gmail.com handling editor: maria rudawska http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.2014.018 mailto:tsurykau%40gmail.com?subject=am.2014.018 250© the author(s) 2014 published by polish botanical society acta mycol 49(2):249–253 tsurykau and czarnota / three lichen species of micarea new to belarus m. denigrata (fr.) hedl., m. erratica, m. lynceola (th. fr.) palice, m. melaena (nyl.) hedl., m. nitschkeana (lahm ex rabenh.) harm., m. peliocarpa (anzi) coppins & r. sant., m. prasina, m. sylvicola (flot.) vězda & v. wirth, m. tuberculata (sommerf.) r.a. anderson [14–16]. however, m. lynceola was reported at the beginning of the 20th century [17] and never confirmed. furthermore, since the specimens of m. prasina were neither tested by tlc nor critically re-examined, this species should be widely treated in the country as m. prasina s.lat. hence, our knowledge of micarea in belarus is still far from complete. during a recent fieldwork in the gomel region in southeastern belarus, three micarea species new to the country were discovered and details of these records are presented here. material and methods the lichen biota of 14 study plots established in 2011–2013 in different types of scots pine forests were investigated; these were selected to represent: cladonia-type, calluna vulgaris-type, vaccinium vitis-idaea-type, pleurozium schreberi-type, pteridium aquilinumtype, vaccinium myrtillus-type, oxalis acetosella-type, polytrichum commune-type and ledum palustre-type stands. the size of each plot was limited by forest sub-blocks and occupied 2–12 ha. in each plot, 10 trees were randomly selected and all lichens were registered within them. secondary chemistry of the sorediate crustose lichens was analyzed by thin-layer chromatography (tlc) in solvent c according to the methods of orange et al. [18]. voucher specimens are deposited in the belarusian polesye scientific herbarium of francisk skorina gomel state university (gsu) and the collection of micarea elachista in the herbarium of the gorce national park (gpn). results and discussion micarea elachista (körb.) coppins & r. sant., bull. br. mus. nat. hist., bot. 11(2): 131 (1983) thallus composed of corticate small warts, pale greenish-grey with numerous immersed or emergent, widely opened pycnidia which are a slightly darker than the thallus. mesoconidia ellipsoid, 3.5–4.5 × 1.2–1.5 µm. apothecia very rare, globose, brown, immarginate with elachista-brown pigment (k+ dissolving and fading into solution) in upper part of the hymenium. ascospores 1-septate, oblong-fusiform, slightly curved, 10–15 × 2–3 µm. no substances detected by tlc. habitat. micarea elachista was found in old-growth vaccinium myrthillus-type pine forest on the bark of two pinus sylvestris l. trees. the sample plot contained some boggy areas with ledum palustre and sphagnum spp. and was untypical of “pinetum myrtillosum” in terms of the degree of humidity. micarea elachista occurred together with chaenotheca spp., chaenothecopsis pusilla (ach.) a.f.w. schmidt, cladonia spp., hypocenomyce scalaris (ach. ex lilj.) m. choisy (often supporting clypeococcum hypocenomycis d. hawksw.), hypogymnia physodes (l.) nyl., lecanora compallens van herk & aptroot, lecidea nylanderi (anzi) th. fr., lepraria spp., micarea denigrata (fr.) hedl., m. melaena (nyl.) hedl. and m. micrococca (körb.) gams ex coppins. 251© the author(s) 2014 published by polish botanical society acta mycol 49(2):249–253 tsurykau and czarnota / three lichen species of micarea new to belarus coppins [19] showed m. elachista to be a predominantly lignicolous species in europe colonizing decorticate trunks or large stumps of old trees, but czarnota [10] has also reported it frequently on the bark of pinus sylvestris in ne poland. general distribution. the lichen is distributed in boreal and temperate europe, occurring in countries neighboring belarus – lithuania [20], poland [10] and adjacent regions of russia [21], as well as in north america [2] and asia [21,22]. specimen examined. belarus, gomel region, gomel district: pribor forest, 1.5 km sw of pribor village, 52°22'n, 30°45'e, 9 oct. 2012, leg. a. tsurykau (gpn/7650). micarea micrococca (körb.) gams ex coppins, checklist of lichens of great britain and ireland: 86 (2002) thallus composed of bright to dull green granules (goniocysts) with numerous immersed, small, white pycnidia producing microconidia 5–7 × 0.8–1 µm. apothecia usually present, whitish, translucent when wet, immarginate, up to 0.3 mm in diam. apothecial section colorless. excipulum strongly reduced. ascospores 1-septate, ovoid to oblong, 8–12 × 3–4 µm. substance detected by tlc: methoxymicareic acid. note. according to phylogenetic analysis, m. micrococca is a polyphyletic taxon. for almost 20 years prior to 2002, this group of taxa was included in m. prasina [23] despite their different chemistry [19]. however, the chemistry appeared to be a sufficiently diagnostic character in further studies and today the presence of methoxymicareic acid is the main feature distinguishing the m. micrococca complex from other members of the m. prasina group. visual identification of m. micrococca s.str. is possible when it is fertile, since its apothecia are white or cream while other representatives of the m. micrococca complex have at least some greyish apothecia due to a trace of sedifolia-grey pigment which turns k± violet and c± violet. the darkest morphs containing methoxymicareic acid may also belong to the recently separated m. byssacea (th. fr.) czarnota, guzow-krzemińska & coppins which probably also occurs in belarus since it is found in neighboring regions [11] (and czarnota, unpublished data). in belarus, both m. micrococca s.str. and m. micrococca s.lat. represent two distinct lineages of the complex [11], but the description, habitat and list of collections below are only given for m. micrococca s.str. habitat. the species was found exclusively on bark of p. sylvestris in three pine forest types, all of which were well-lit with a sufficient amount of moisture, ranging from the wet polytrichum-type to the medium humid pleurozium schreberi-type. general distribution. it is difficult to evaluate the true world distribution of m. micrococca s.str. as representatives of the m. micrococca complex were critically revised only recently [11], but it has been reported so far from the czech republic, estonia, finland, germany, lithuania, poland, slovenia and switzerland [11,24]. however, this corticolous lichen-forming fungus inhabiting acidic bark appears to be frequent in europe especially within large more or less managed coniferous woodlands. specimens examined. belarus, gomel region, gomel district: kalinino forest, 1.5 km e of tereshkovichi village, 52°15'n, 30°59'e, 3 aug. 2011, leg. a. tsurykau (gsu/1867); same forest, 1.3 km ne of tereshkovichi village, 52°15'n, 30°58'e, 2 oct. 2012, leg. a. tsurykau (gsu/1868); pribor forest, 0.5 km s of pribor village, 52°23'n, 30°47'e, 10 oct. 2012, leg. a. tsurykau (gsu/1869); same forest, 1.5 km sw of pribor village, 52°22'n, 30°45'e, 10 oct. 2012, leg. a. tsurykau (gsu/1870). 252© the author(s) 2014 published by polish botanical society acta mycol 49(2):249–253 tsurykau and czarnota / three lichen species of micarea new to belarus micarea misella (nyl.) hedl., bih. k. svenska vetensk akad. handl. 18(3): 78 (1892) thallus inconspicuous, partially on pine bark as a thin algal film, and partially on the thallus of m. denigrata. pycnidia numerous, black, stalked, containing sedifolia-grey pigment, k+ violet, c+ violet. mesoconidia simple, cylindrical, 5.0–5.5 × 1.4–1.6(–1.9). apothecia absent in belarusian material. no substances detected by tlc. habitat. the species was collected in young vaccinium vitis-idaea-type pine forest growing on the edges of bark plates in bark fissures of scots pine. kotlov [25] noted that micarea misella is exclusively lignicolous in russia, and coppins [26] noted that it is almost always lignicolous and rarely grows on the bark of old trees. general distribution. micarea misella is reported from europe and north america, as well as from south america [2] and the asian part of russia [21]. amongst neighboring countries, it occurs in lithuania [20], ukraine [27] and in adjacent regions of russia [21], and in poland [10] it is one of the commonest representatives of the genus. specimen examined. belarus, gomel region, gomel district: staro-djatlovichskoje forest, 2 km sw of staryje djatlovichi village, 52°13'n, 30°49'e, 1 aug. 2013, leg. a. tsurykau (gsu/1763). acknowledgments we are very grateful to the two anonymous reviewers for valuable comments on the manuscript, and especially to professor mark seaward (bradford) for his linguistic corrections and other improvements. authors’ contributions the following declarations about authors’ contributions to the research have been made: field research and draft of the manuscript: at; species identification and critical revising: pc; final writing of the manuscript: at, pc. references 1. fries em. systema orbis vegetabilis. lund: 1825. 2. coppins bj. micarea fr. 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(vol 2 pt 3). http://dx.doi.org/10.1639/0007-2745-113.2.333 http://dx.doi.org/10.1017/s0024282911000338 http://dx.doi.org/10.5248/116.61 http://dx.doi.org/10.13158/heia.26.1.2013.201 http://dx.doi.org/10.1017/s0024282999000730 http://dx.doi.org/10.1017/s0024282999000730 abstract introduction material and methods results and discussion micarea elachista (körb.) coppins & r. sant., in coppins, bull. br. mus. nat. hist., bot. 11(2): 131 micarea micrococca (körb.) gams ex coppins, in coppins, checklist of lichens of great britain and ir micarea misella (nyl.) hedl., bih. k. svenska vetensk akad. handl. 18(3): 78 (1892) acknowledgments authors’ contributions references 2014-12-31t17:37:13+0000 polish botanical society 2014-09-03t19:46:24+0200 polish botanical society 2014-09-03t19:46:59+0200 polish botanical society 2014-09-06t20:25:20+0200 polish botanical society 2014-08-31t21:55:53+0200 polish botanical society 2014-09-03t19:47:13+0200 polish botanical society 2014-09-02t20:10:44+0200 polish botanical society 2014-09-05t16:23:30+0200 polish botanical society 2014-09-06t20:22:38+0200 polish botanical society 2014-09-08t11:11:17+0200 polish botanical society 2014-08-31t21:54:44+0200 polish botanical society 2014-09-02t20:11:06+0200 polish botanical society 2014-09-05t16:23:09+0200 polish botanical society 2014-09-05t16:24:22+0200 polish botanical society 2014-09-02t20:09:50+0200 polish botanical society 2014-08-31t21:56:56+0200 polish botanical society 2014-08-31t21:56:27+0200 polish botanical society 2014-09-05t16:24:39+0200 polish botanical society 2014-09-06t20:22:53+0200 polish botanical society 2014-09-06t20:25:02+0200 polish botanical society microfungi of carpinus betulus from poland i. annotated list of microfungi andrzej chlebicki1 and maria a. chmiel2 1department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl 31 512 kraków, ibchlebicki@ib pan.krakow.pl 2department of botany and mycology, maria curie skłodowska university akademicka 19, pl 20 033 lublin c h l e b i c k i a., c h m i e l m. a.: microfungi of carpinus betulus from poland. i. annotated list of microfungi. acta mycol. 41 (2): 253 278, 2006. the compiled microfungi list comprises 115 taxons noted in poland, of them, 28 parasitic. 10 species of microfungi were host specialized (exclusive, or partially exclusive for hornbeam). key words: hornbeam fungi, list of species, mycogeography, poland introduction the majority of hornbeam species was reported from eastern asia. thus the distribution centrum of the genus carpinus l. is located in central china (b o r a t y ń s k a 1993; c h e n et al. 1999). it is difficult to establish a precise number of all hornbeam species. until recently over 80 separated species have been reported. according to b o r a t y ń s k a (1993) only 21 species can be referred to the genus. the residual taxa are questionable. at present from europe are known two species: carpinus betulus l. and c. orientalis mill. the hornbeam species are distributed in three regions of the northern hemisphaere: north america (1 species), europe and asia minor (2 species) and eastern asia (18 species). the genus carpinus supports many microfungi, some of them are confined to the genus or related genera. there are some articles devoted fungi on carpinus betulus e.g. fa k i r o v a (1993), and two other hornbeam species such as c. virginiana (marshall) sudw. (fa r r et al. 1989; b i l l s , p o l i s h o o k 1991) and c. cordata blume (va s i l y e v a 1998). these plants are partially representative for the three mentioned disjunctive regions. however mycological research of the most interesting region in central china is largely neglected. we found only single report of melampsoridium carpini (nees) dietel on carpinus fargesiana h. winkler from sichuan in china (z h a n g et al. 1997). finds on carpinus cordata from russian far east (va s i l y e v a 1998) showed that we can expect much more exclusive species of microfungi from this region. microfungi noted on carpi acta mycologica vol. 41 (2): 253-278 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 254 a. chlebicki and m. a. chmiel nus virginiana and c. cordata have only little biogeographical significance, because of fragmentary knowledge of their spatial distribution pattern. mycological data of the hornbeam tree in polish territoty are fragmentary. wo j e w o d a (2003) noted 129 macromycetes collected by various authors on the host plant, among them 27 parasitic fungi and 102 species of saprotrophs. under the tree has been noted 82 species of fungi growing in the soil (wo j e w o d a 2003). microfungi are insuficiently investigated. only białowieża n. p. and kampinos n. p. are more detaily studied. other areas posses mostly scarcely data. the listed below microfungi have been collected by many mycologists. in poland these fungi were collected by s c h r o e t e r (1908), e i c h l e r (1902, 1904, 1907), r o u p p e r t (1909), n a m y s ł o w s k i (1909), s t e c k i (1910), wa ś n i e w s k i (1911), s i e m a s z k o (1923), tr z e b i ń s k i e t a l . (1916), wr ó b l e w s k i (1918, 1925), d o m i n i k (1936), s k i r g i e ł ł o (1960), tr u s z k o w s k a (1959, 1960, 1965), g o ł ą b (1978), b o r o w s k a (1966, 1986, 1987, 1989), s a ł a t a (1972, 1975), we b e r c z e r w i ń s k a (1974), tr u s z k o w s k a & c h l e b i c k i (1983), c h l e b i c k i (1986, 1991, 2002, 2005a, 2005b, 2006), m u ł e n k o (1996), b u j a k i e w i c z et al. (1997), m a ń k a et al. (2002) and p i ą t e k (2004). the compiled list of microfungi comprises 115 taxons, of them, 28 parasitic species. 10 species of microfungi were host specialized (exclusive species, or partially exclusive species for hornbeam). gnomoniae fimbriata (pers.) fuckel appear the most common hornbeam ascomycete in poland. there are also collected very rare species such as camarops plana pouzar, camarops polysperma (mont.) j. h. mill., lasiosphaeria hirsuta (fr.) ces. et de not., lophiostoma curreyi sacc., nemania atropurpurea (fr.) pouzar, oidium carpini foitzik and others. presence of following exclusive species was noted in polish territory: camarops plana, diaporthe carpini (pers.) fuckel, encoelia carpini (rehm) boud., gnomonia fimbriata, massaria carpinicola tul. et c. tul., melampsoridium carpini, melanconis spodiaea tul. et c. tul., oidium carpini, pezicula carpinea (pers.) tul. ex fuckel and phyllosticta carpini schulzer et sacc. list of species abbreviations: coll. collected; distr. district; var. variety; subsp. subspecies; n. p. national park; r.p. regional park; unpubl. unpublished; synonymes and doubtful taxa are designated by italic letters; parasitic species are marked by ♦. alysidium resinae (fr.) m. b. ellis, anamorphic fungus, botryobasidium, basidiomycetes. locality: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead wood of betula pendula, carpinus betulus, quercus robur (b o r o w s k a l. c.). ♦ apiosporopsis carpinea (fr.) traverso, syn.: sphaerognomonia carpinea (fr.) potiebnia, sphaeria carpinea fr.: fr., laestadia carpinea (fr.: fr.) sacc. guignardia carpinea (fr.) j. schroet., gnomoniella carpinea (fr.) monod, anamorph: monostichella robergei (desm.) hoehn., diaporthales. microfungi of carpinus betulus 255 localities: teleomorph: zielona góra, wołów, trzebnica, wrocław (s c h r o e t e r 1908), kampinos n. p., dębina reserve (b o r o w s k a 1966), białowieża n. p. (c h l e b i c k i et al. 1996); anamorph: białowieża n. p. (m u ł e n k o 1996). host: on living leaves (teleomorph on overwintered leaves) of acer, alnus, betula, carpinus betulus, c. caroliniana, castanea, corylus and ostrya and quercus (c o n n e r s 1967; b a r r 1978; fa r r et al. 1989). the species was noted in europe and north america (c o n n e r s l. c.; fa r r et al. l. c.; fa k i r o v a 1993). barrmaelia oxyacanthae (mont.) rappaz, syn.: anthostomella melanotes (berk. et br.) p. m. d. martin, xylariales. locality: lower silesia, środa śląska near wrocław (s c h r o e t e r 1908). host: on dead wood of ribes petreum, populus tremula, carpinus betulus (s c h r o e t e r l. c.), salix cinerea (c h l e b i c k i , unpubl. data). earlier its localities were referred to anthostomella melanotes (berk. et br.) p. m. d. martin. bertia moriformis (tode) de not., sordariales. localities: białowieża n. p. (c h l e b i c k i et al. 1996). host: on decorticated wood of stumps and branches of alnus, fagus, picea, tilia, acer and pinus (c h l e b i c k i 1991; b u j a k i e w i c z et al. 1997). both varieties of the species: bertia moriformis var moriformis as well as b. moriformis var. latispora were found on hornbeam wood in białowieża n. p. (c h l e b i c k i l. c.). biscogniauxia repanda (fr.) kuntze, xylariales. localities: byk near międzyrzec, on carpinus betulus (e i c h l e r 1907). host: on dead branches of sorbus aucuparia, sorbus aria (p o u z a r 1986a). c h l e b i c k i and b u j a k i e w i c z (1994) and c h l e b i c k i (2005b) cited some finds from poland on sorbus aucuparia. g r a n m o et al. (1989) cited also betula, prunus, malus, quercus, alnus and tilia as rare host plants. p o u z a r (l. c.) considered sorbus aucuparia as the most important host. j u et al. (1998) mentioned two finds from ussuriysky reserve on malus mandshurica, however, va s i l y e v a (1998) described it as separate taxon, biscogniauxia mandshurica which differs from b. repanda by narrower ascospores and umbilicate ostioles. e i c h l e r (l. c.) reported the species as nummularia repanda (fr.) nitschke with ascospores 10-12 x 5-6 um. (b. repanda: 10-15 x 4-6 um; b. nummularia: 10-16 x 6-10 um). va s i l y e v a (1988) noted related species biscogniauxia succenturiata (tode: fr.) o. kuntze on carpinus cordata from russian far east. j u et al. (l. c.) revised the type material of sphaeria succenturiata tode from ups and considered that it is lopadostoma gastrinum, but the descriptions of persoon, fries and nitschke suggest a biscogniauxia taxon. b. repanda is known from europe, asia and north america (g r a n m o et al. 1989; j u et al. l. c.; va s i l y e v a l. c.). bispora betulina (corda) s. hughes, anamorphic fungus, incertae sedis, ascomycetes. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead wood of carpinus betulus, quercus robur, betula pendula, tilia cordata (b o r o w s k a 1987), carpinus betulus, betula pendula, fagus orientalis, pyrus sp., populus sp., quercus sp. (m e l n i k 2000). it is common species, known from europe and north america (e l l i s 1971). 256 a. chlebicki and m. a. chmiel bolinia tubulina sacc. camarops tubulina. note of this species (tr u s z k o w s k a 1965) refers to camarops polysperma. brachysporium nigrum (link) s. hughes, anamorphic fungus, incertae sedis, ascomycetes. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum (b o r o w s k a 1987); jura krakowska, prądnik (s c h e u e r , c h l e b i c k i 1997). host: on rotting wood of carpinus betulus (belorussia), betula, corylus, populus tremula (m e l n i k 2000). cacumisporium capitulatum (corda) s. hughes, anamorphic fungus, incerte sedis. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead wood of carpinus betulus. camarops tubulina (alb. et schw.: fr.) shear, xylariales. the material from białowieża n. p. reported as bolinia tubulina sacc. (tr u s z k o w s k a 1965) is erroneously determined and in fact belongs to camarps polysperma (n a n n f e l d t 1972). camarops tubulina was noted only on wood of picea abies, abies alba (n a n n f e l d t l. c.; h i l b e r , h i l b e r 1980) and occasionally on fagus sylvatica (h o l e c 2005). camarops plana pouzar, boliniales, ascomycetes (fig. 1). locality: poland: białowieża n. p., tilio-carpinetum, on trunk of carpinus betulus, lying in ditch, august 1961, coll.: w. truszkowska. host: on dead wood of trunk of carpinus betulus (p o u z a r 1986). the species has been described by p o u z a r (1986b) on the basis of material from slovakia. tr u s z k o w s k a (1965) reported the species from białowieża n. p. as camarops tubulina (alb. et schwein.) shear, see c h l e b i c k i (2006). fig. 1. section of stroma of camarops plana pouzar; according to drawing of ewa szumińska. microfungi of carpinus betulus 257 camarops polysperma (mont.) j. h. mill., boliniales. locality: białowieża n. p. (tr u s z k o w s k a 1965). host: noted on dead branches and trunks of alnus glutinosa, a. incana, acer sp., fagus sylvatica, carpinus betulus (n a n n f e l d t 1972), citrus sp., magnolis sp., quercus sp., scordophloeus zenkeri (h i l b e r , h i l b e r 1980), and fraxinus excelsior (c h l e b i c k i unpubl. data). it is a rare species, restricted to the old forests in north poland and carpathians (c h l e b i c k i , b u j a k i e w i c z 1994), known from northern hemisphere (fa r r et al. 1989), argentina (m e r c u r i 1972), europe, north america, southern america, africa (h i l b e r , h i l b e r l. c.) and east china (a b e , l i u 1995). some other species of the genus camarops have been noted on hornbeam. camarops pugillus (schw.: fr.) shear was noted in czech and sweden (l u n d q v i s t 1987) as well in austria (s c h e u e r 1997). camarops microspora (p. karst.) shear, mostly noted on alnus, was noted on hornbeam branches in czech (p o u z a r 1986b) and germany (n a n n f e l d t l. c.). cenangium carpini rehm encoelia carpini. ceratostomella ampullasca (cooke) sacc. endoxyla cirrhosa. chaetopsis grisea (ehrenb.) sacc., anamorphic fungus, incertae sedis, ascomycetes. localities: dębina reserve, kampinos n. p. (b o r o w s k a 1966), białowieża n. p. (b o r o w s k a 1986). host: noted on dead bark and wood, rarely on old leaves of acer, alnus, carpinus betulus, fagus, laurus, quercus, salix, ulmus (b o r o w s k a l. c.). it is species known from europe. chaetosphaeria ovoidea (fr.) constant., k. holm et l. holm, syn.: chaetosphaeria glauca hol.-jech., anamorph: menispora glauca pers., trichosphaeriales. localities: anamorph is a common fungus in all parts of poland (b o r o w s k a 1986). host: noted on bark and wood of acer, carpinus betulus, betula, quercus, fagus, populus (in poland). teleomorph was found on wood of fagus sylvatica and quercus sp. in czech republic and on populus tremula in lithuania (tr e i g i e n ë , m a r k o v s k a y a 2003). chaetosphaeria inaequalis (grove ex berl. et voglino) w. gams et hol.-jech., anamorph: gonytrichum caesium c. g. nees et t. f. l. nees var. caesium, trichosphaeriales. localities: kampinos n. p., mostly noted on carpinus betulus and quercus sp. (b o r o w s k a 1986), known from europe and north america (b o r o w s k a l. c.). host: on dead wood and bark of abies, acer, alnus, betula, buxus, carpinus betulus, corylus, robinia, salix, taxus, ulmus, ilex. chaetosphaeria innumera berk. et broome ex tul. et c. tul., anamorph: chloridium botryoideum (corda) s. hughes var. botryoideum, trichosphaeriales. localities: kampinos n. p. (b o r o w s k a 1986). host: on bark of dead plant of carpinus betulus, fagus, alnus, betula, quercus, salix, sorbus, ulmus, tilia (b o r o w s k a l. c.). the species was noted in germany, sweden, hungary, great britain and canada (b o r o w s k a l. c.). chaetospheria lentomita w. gams et hol.-jech., anamorph: chloridium pachytrachelum w. gams et hol.-jech., trichosphaeriales. 258 a. chlebicki and m. a. chmiel locality: świętokrzyski n. p. (b o r o w s k a 1986). host: on dead wood of.fagus, carpinus, quercus, pinus (b o r o w s k a l. c.). chaetosphaeria myriocarpa (fr.) c. booth, anamorph: chloridium clavaeforme (preuss) w. gams et hol.-jech., trichosphaeriales. localities: anamorph: common in all part of poland, kampinos n. p., krzywa góra, sieraków, zamczysko. tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1986, 1987); teleomorph: jura krakowska, prądnik (s c h e u e r , c h l e b i c k i 1997). host: on rotting wood of various trees also on carpinus betulus (d e n n i s 1968). anamorph has been noted on acer, alnus, amygdalus, betula, carpinus, castanea, cedrus, corylus, crataegus, fagus, fraxinus, pinus, sambucus, sorbus, quercus and ulmus (b o r o w s k a l. c.). the species is known from europe, north america, asia and new zeland (b o r o w s k a l. c.). chaetosphaeria preussii w. gams et hol. -jech., anamorph: chloridium preussii w. gams et hol.-jech., trichosphaeriales. localities: anamorph: kampinos n. p. (b o r o w s k a 1987), known also from europe and canada (b o r o w s k a 1986). host: on dead wood of acer, carpinus betulus, fraxinus, quercus (b o r o w s k a l. c.). chalara breviclavata nag raj et w. b. kendr., anamorphic fungus, incertae sedis, ascomycetes. localities: białowieża n. p., kampinos n. p. (b o r o w s k a 1986). host: on dead wood of betula, carpinus betulus, quercus and tilia (b o r o w s k a l. c.), fraxinus (n a g r a j , k e n d r i c k 1975). the species is known from europe and canada (b o r o w s k a l. c.). chalara cylindrosperma (corda) s. hughes, anamorphic fungus, incertae sedis, ascomycetes. localities: białowieża n. p. (b o r o w s k a 1986). host: on dead stems and fruits of aconitum, aesculus, agathis, betula, fagus, gleditschia, ilex, picea, pinus, podocarpus, tilia, carpinus (b o r o w s k a l. c.). the species was noted in europe, canada, india and new zeland (b o r o w s k a l. c.). chalara insignis (sacc., m. rousseau. et e. bommer) s. hughes, hyphomycetes. localities: warsaw (b o r o w s k a 1986). host: on dead wood and bark of corylus, picea, carpinus, poria and quercus (b o r o w s k a l. c.). it was earlier reported from great britain and canada. chloridium clavaeforme vide teleomorph chaetosphaeria myriocarpa. chloridium preussii w. gams et hol.-jech., vide teleomorph chaetosphaeria preussii. cheirospora botryospora s. hughes, syn.: thyrsidium botryosporum sacc., anamorphic fungus, ascomycetes. localities: białowieża n. p. (tr u s z k o w s k a 1965). host: plurivorous species, reported on dead twigs of cornus alba, hedera helix, fagus grandifolia, quercus robur in u. k., austria and u.s.a. (s u t t o n 1980), alnus glutinosa and betula pubescens ssp. carpatica in poland (c h l e b i c k i et al. 1996), on carpinus orientalis in caucasus, georgia (m e l n i k 2000) and carpinus betulus in lithuania (i g n a t a v i c i û t ë , tr e i g i e n ë 1998). microfungi of carpinus betulus 259 chloridium virescens var. caudigerum (höhn.) w. gams et hol. jech., syn.: cirrhomyces caudigerus höhn., anamorphic fungus, teleomorph: chaetosphaeria, sordariales. localities: b o r o w s k a (1986) mentioned this species on wood of hornbeam from poland. it occurs in europe, north america, africa and sri lanka (b o r o w s k a l. c.). host: on dead wood of acer, alnus, betula, carpinus betulus, fagus, picea, quercus, populus. chloridium virescens var. chlamydosporum (j. f. h. beyma) w. gams et hol. jech., anamorphic fungus, teleomorph: chaetosphaeria, sordariales. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead bark and wood and overwintered leaves of aesculus, alnus, carpinus, cedrus, fagus, fraxinus, populus, quercus, salix, sorbus, spirea, dead rots of picea and caragana, dead stems of urtica and juncus (b o r o w s k a 1986). it is the most common species of chloridium in poland (b o r o w s k a l. c.). the species was noted in europe, asia, north america, africa, australia and new guinea (b o r o w s k a l. c.). cladosporium cladosporioides (fresen) g. a. de vries, anamorphic fungus, teleomorph: mycosphaerella, mycosphaerellales. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-quercetum (b o r o w s k a 1987). host: it is cosmopolitan species, known on dead wood of vide variety of plants. coniochaeta ligniaria (grev.) massee, sordariales. localities: białowieża n. p. (tr u s z k o w s k a 1965), known from europe (a r x , m ü l l e r 1954), temperate northern hemisphere (fa r r et al. 1989). host: on corticated and decorticated branches as well as on herbaceous stems of abies, acer, alnus, ceanothus, cercocarpus, crataegus, larix, oplopanax, ostrya, physocarpus, pinus, populus, sambucus, tsuga (fa r r et al. l. c.). cordana pauciseptata preuss, anamorphic fungus, incertae sedis, ascomycetes. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead wood of decorticated twigs of alnus incana, carpinus betulus (in lithuania), fagus orientalis, rosa spp., populus sp. (m e l n i k 2000). b o r o w s k a (l. c.) noted it on carpinus betulus, quercus robur and betula pendula in poland. ♦ cryphonectria gyrosa (berk. et broome) sacc. et d. sacc, syn.: endothia gyrosa (schwein.: fr.) fr., diaporthales. locality: strzelińskie hills (tr u s z k o w s k a , c h l e b i c k i 1983). host: on twigs of acer, castanea, corylus, fagus, ilex, liquidambar, quercus, ulmus, vitis (fa r r et al. 1989). it was reported from north america and australia (fa r r et al. l. c.). cryptosporella aurea fuckel melanconis xanthostroma. cryptosporiopsis fasciculata vide teleomorph: pezicula carpinea. ♦ cylindrocarpon magnusianum (sacc.) wollenw, teleomorph: nectria, sordariales. locality: wronczyn near poznań (m a ń k a et al. 2002). host: on living roots of carpinus betulus. 260 a. chlebicki and m. a. chmiel cylindrotrichum oligospermum (corda) bonord., anamorphic fungus, teleomorph: chaetosphaeria, sordariales. locality: białowieża n. p. (b o r o w s k a 1986). host: on dead wood and stems of betula, carpinus, ulmus, umbelliferae (b o r o w s k a l. c.). it is the most common species of cylindrotrichum. ?cytospora decipiens sacc. (tr u s z k o w s k a , c h l e b i c k i 1983), it is probably cytospora leucosperma (pers.) fr. cytospora decorticans sacc., białowieża n. p. (tr u s z k o w s k a 1959). dasyscyphus fuscescens (pers.) gray lachnum fuscescens var. fuscescens. diaporthe betuli (pers.) winter diaporthe carpini. ♦ diaporthe carpini (pers.) fuckel, syn.: diaporthe betuli (pers.) winter, anamorph: discosporina deplanata (höhn.) höhn., diaporthales. localities: zielona góra, legnica, wrocław, niemcza, wałbrzych (s c h r o e t e r 1908), warszawa (k o c h m a n 1964), białowieża n. p. (tr u s z k o w s k a 1965), strzelińskie hills (tr u s z k o w s k a , c h l e b i c k i 1983). host: on senescent twigs of carpinus betulus (m u n k 1957), rarely corylus avellana (we h m e y e r 1933) and c. mandshurica (va s i l y e v a 1998). it was noted in europe (fa k i r o v a 1993) and russian far east (va s i l y e v a 1989). ♦ diaporthe eres nitschke syn.: diaporthe sordida nitschke, diaporthales. locality: zielona góra (s c h r o e t e r 1908). host: on senescent and dead twigs and branches, plurivorous species, noted on 65 host plant species (we h m e y e r 1933; fa r r et al. 1989) from temperate areas of northern hemisphaere. diaporthe bitorulosa (berk. et broome) sacc. wusteneia xanthostroma. diaporthe hyperopta nitschke f. major wehm. melanconis chrysostroma. diaporthe sordida nitschke = diaporthe eres. ♦ diatrype decorticata (pers.) rappaz, xylariales. localities: białowieża n. p. (c h l e b i c k i et al. 1996), puszcza augustowska forest, choszczewo (c h l e b i c k i unpubl. data). the species was noted in europe (r a p p a z 1987). host: on dead twigs of carpinus, fagus, tilia, crataegus, prunus, sali (r a p p a z l. c.). diatrype flavovirens (pers.) fr., xylariales. localities: międzyrzec (e i c h l e r 1907), białowieża n. p. (c h l e b i c k i et al. 1996), puszcza augustowska forest, dworczysko, near czarna hańcza river (c h l e b i c k i , unpubl. data). the species was reported from europe, north america, africa, asia (r a p p a z 1987). host: on dead twigs and branches of carpinus, prunus spinosa, sambucus callicarpa, s. racemosa, acer pseudoplatanus, quercus pubescens, hedera helix, viburnum opulus, cornus sp., corylus avellana, fagus sylvatica, fraxinus excelsior, cydonia vulgaris, prunus armeniaca, salix sp. and tilia sp. (r a p p a z l. c.). ♦ diatrype stigma (hoffm.) fr., xylariales. localities: chirkowa near świecie (h e n n i n g s 1892), białowieża n. p. (tr u s z k o w s k a 1959, 1965, c h l e b i c k i et al. 1996), puszcza borecka forest, boczki reserve (c h l e b i c k i , k r z y ż a n o w s k a 1995), puszcza augustowska forest, czerwone bagno reserve, bagno ławki (c h l e b i c k i , unpubl. data). host: on dead twigs and branches of quercus, rosaceae, betulaceae (r a p p a z 1987), quercus microfungi of carpinus betulus 261 acutissima (a b e 1986). it is known from europe and north america (r a p p a z l. c.; fa k i r o v a 1993) and japan (a b e 1986). ♦ diatrype subaffixa var. rappazii chleb., xylariales (fig. 2). localities: białowieża n. p., puszcza romincka forest, boczki reserve puszcza borecka forest, czerwony dwór (c h l e b i c k i , k r z y ż a n o w s k a 1995; c h l e b i c k i et al. 1996; c h l e b i c k i , unpubl. data). host: on dead wigs and branches of carpinus betulus (c h l e b i c k i , k r z y ż a n o w s k a l. c.). diatrype subaffixa (schwin.) cooke is known from type locality in new jersey (usa) on rosaceae (r a p p a z 1987; c h l e b i c k i , k r z y ż a n o w s k a l. c.). fig. 2. section of stroma of diatrype subaffixa var. rappazii chlebicki: a pulvinate stroma kramf41521; b undulate stroma kramf41577; c ascospores, some with budding apex. 262 a. chlebicki and m. a. chmiel ♦ diatrypella favacea (fr.) ces. et de not., syn.: d. verrucaeformis (fr.) nitschke, xylariales. localities: strzelińskie hills (tr u s z k o w s k a , c h l e b i c k i 1983), białowieża n. p. (tr u s z k o w s k a 1959, 1965; c h l e b i c k i 1986; c h l e b i c k i et al. 1996), puszcza romincka forest, boczki reserve (c h l e b i c k i 2005a). host: on senescent twigs and branches of alnus, betula, corylus, carpinus (c r o x a l l 1950; c h l e b i c k i l. c.). c r o x a l l (1950) and c h l e b i c k i (l. c.) considered d. verrucaeformis as synonyme of d. favacea. discosporina deplanata (höhn.) höhn., vide teleomorph: diaporthe carpini. enchnoa infernalis (kunze) fuckel, sordariales. locality: lower silesia, świdnica (s c h r o e t e r 1908). host: on dead twigs and branches of quercus (m u n k 1957; d e n n i s 1968), carpinus betulus (s c h r o e t e r l. c.), salix caprea, s. myrsinifolia (m a t h i a s s e n 1993). the fungus was reported from great britain (d e n n i s l. c.), danmark (m u n k l. c.), norway (m a t h i a s s e n 1989, l. c.) and sweden (e r i k s s o n 1992). similar species, enchnoa subcorticalis (peck) barr was noted on branches of carpinus caroliniana (fa r r et al. 1989). ♦ encoelia carpini (rehm) boud., syn.: cenangium carpini rehm, helotiales. localities: międzyrzec (e i c h l e r 1902), białowieża n. p. (a n o n i m 1968; k o t l a b a , l a z e b n i č e k 1967). host: on senescent twigs of carpinus betulus. related species, encoelia furfuracea (roth ex pers.) p. karst. inhabit genera from botyloideae and coryloideae: alnus and corylus in great britain (d e n n i s 1968). k u t o r g a (1989) found e. furfuracea on decorticated wood and bark of alnus in lithuania. these relationships can indicate that fungi from the genus encoelia coevolved with various genera of betulaceae from tertiary. endothia gyrosa (schwein.: fr.) fr. cryphonectria gyrosa. endoxyla cirrhosa (pers.) e. müll. et arx, syn.: ceratostomella ampullasca (cooke) sacc., xylariales. locality: jura krakowska, prądnik (s c h e u e r , c h l e b i c k i 1997). host: noted on rotting wood of acer, carpinus betulus, quercus (d e n n i s 1968; s c h e u e r , c h l e b i c k i l. c.). eutypa lata (pers.) tul. et c. tul., diatrypales, noted on decorticated wood. localities: puszcza romincka forest, boczki reserve (c h l e b i c k i , unpubl. data), noted also on acer platanoides, near hańcza lake. host: it is plurivorous and common species (r a p p a z 1987). eutypa lejoplaca (fr.) cooke, xylariales. locality: białowieża n. p. (c h l e b i c k i et al. 1996). host: on dead branches of acer monspessulanus, a. platanoides, a. pseudoplatanus (r a p p a z 1987), carpinus betulus (c h l e b i c k i et al. l. c.). the species was noted in europe (r a p p a z l. c.). eutypella cerviculata (fr.) sacc., xylariales. localities: lower silesia, borowa oleśnicka coll.: r. guziak, 1991, kram 41372; turew near poznań, coll.: w. truszkowska, noted also in białowieża n. p. (tr u s z k o w s k a 1959, c h l e b i c k i et al. 1996). host: on dead twigs and branches of alnus sp., alnus incana, a. tenuifolia, carpinus betulus, carpinus americana, betula nigra, betula sp. (r a p p a z 1987), amelnachier, corylus, ostrya, prunus and quercus (fa r r et al. 1989). it is known in temperate areas of norhtern hemispahere (fa r r et microfungi of carpinus betulus 263 al. l. c.). va s i l y e v a (1998) reported eutypella corynostomoides (rehm) rappaz on branches of carpinus cordata from russian far east. ♦ gibberella baccata (wallr.) sacc., hypocreales. localities: oława, lower silesia (s c h r o e t e r 1908). it is cosmopolitan species. host: cankering of woody plants such as carpinus, citrus, cotoneaster, ficus, juglans, malus, morus, prunus, pyrus, robinia (fa r r et al. 1989), s c h r o e t e r (l. c.) noted it on robinia pseudoacacia, viburnum opulus and carpinus betulus. glonium lineare (fr.) de not., dothideales. localities: lower silesia: dobrogoszcz,, wrocław, zyrowa near strzelce opolskie, st. anna mt. (s c h r o e t e r 1908). host: plurivorous species, noted on dead wood and branches, also on carpinus, liquidambar, fagus, quercus and vitis (s c h r o e t e r l. c., fa r r et al. 1989). gnomonia fimbriata (pers.) fuckel syn.: mamiania fimbriata (pers.: fr.) ces. et de not., anamorph asteroma carpini (libb.) sutton, diaporthales. localities: klęczany (n a m y s ł o w s k i 1909), międzyrzec (e i c h l e r 1907), ludwikowo near poznań, zagórzek near gdynia, nieporuszewo near grodzisk, miradz near strzelno (d o m i n i k 1936) czerwieńsk, złotoryja, jawor, wrocław, brzeg, strzegom, krasków near świdnica, niemcza, strzelin, ziębice, kamieniec near ząbkowice, dzierżoniów, nowa ruda, opole, goszczowiec near niemodlin, zakrzów near strzelce opolskie (s c h r o e t e r 1908), kuczek near ciechocinek (r u p p e r t 1909), łysa góra (wa ś n i e w s k i 1911), białowieża n. p. (s i e m a s z k o 1923; tr u s z k o w s k a 1959, 1965; m u ł e n k o l. c.), sikornik near kraków (wr ó b l e w s k i 1925), wołtuszowa near rymanów zdrój (s t e c k i 1910), anamorph. locality: anamorph: białowieża n. p. (m u ł e n k o 1996). host: on leaves of carpinus betulus, carpinus cordata (va s i l y e v a 1998), ostrya virginiana (b a r r 1978). the fungus is known from europe, russian far east (va s i l y e v a l. c.) and north america (b a r r l. c.). gnomonia setacea (pers.) ces. et de not., diaporthales. localities: chełm dolny, środa śląska, lower silesia (s c h r o e t e r 1908). host: on overwintered leaves of various species (b a r r 1978), in poland noted on fagus sylvatica, quercus robur, betula pendula, b. pubescens, alnus glutinosa, corylus avellana (s c h r o e t e r l. c.), cupules of quercus and twigs of alnus (c h l e b i c k i et al. 1996). it is cosmopolitan species, b a r r (l. c.) noted this fungus on betula, alnus, quercus, castanea and occasionally on lycopodium. gnomoniella carpinea (fr.) monod sphaerognomonia carpinea. guignardia carpinea (fr.) j. schroet. sphaerognomonia carpinea. gonytrichum caesium var. chloridioides w. gams et hol.-jech., teleomorph: chaetosphaeria, sordariales. locality: kampinos n. p. (b o r o w s k a 1986). host: on dead wood of betula, carpinus, fagus, quercus and tilia and rarely isolated from the soil (b o r o w s k a l. c.). it was noted in france, niderland, great britain and czechoslovakia (b o r o w s k a l. c.). ♦ hapalocystis bicaudata fuckel, syn.: melanconis berkeleyi tul., anamorph: stilbospora macrosperma pers. ex mérat., diaporthales. 264 a. chlebicki and m. a. chmiel localities: czerwieńsk in zielona góra region, wrocław, as pseudovalsa macrosperma tulasne (s c h r o e t e r 1908), białowieża n. p. (tr u s z k o w s k a 1965; c h l e b i c k i 1991), wierzchlas (we b e r c z e r w i ń s k a 1974), śnieżnik mt. in sudety mts. (c h l e b i c k i l. c.). host: on senescent twigs and branches of trees. g r o v e (1937) and s u t t o n (1980) gave as substratum carpinus betulus, fagus sylvatica, ulmus sp. and cornus sp. c h l e b i c k i (l. c.) found it on carpinus betulus and ulmus montana. helicoon richonis (boud.) linder, incerte sedis, ascomycetes. locality: kampinos n. p., sieraków (b o r o w s k a 1989), reported from europe and canada (b o r o w s k a l. c.). host: on rooten wood of alnus glutinosa, betula sp., pinus sylvestris, carpinus betulus, and cone of picea abies as well as decaying wood of quercus, populus, salix, fallen leaves and fruits of these plants (b o r o w s k a l. c.). helicoon sessile morgan, incerte sedis, ascomycetes. localities: kampinos n. p., sieraków (borowska 1989), known from great britain, poland and usa (b o r o w s k a 1989). host: on rotten wood of acer, quercus, fagus, carpinus betulus and coniferous wood (e l l i s , e l l i s 1986; b o r o w s k a l. c.). helicosporium vegetum nees, see tubeufia cerea. helotium phyllophilum (desm.) karst. hymenoscyphus phyllophilus. helotium fructigenum(bull.: merat) karsten hymenoscyphus fructigenus. hymenoscyphus epiphyllus (pers.) rehm ex kauffman, helotiales. localities: dębina reserve (b o r o w s k a 1966), łęczna-włodawa lake district, lake długie (c h m i e l 1987, 1990), roztoczański n. p. (s a ł a t a 1972). host: on fallen leaves of various plants,it is common and plurivorous species (d e n n i s 1968). hymenoscyphus fructigenus (bull.) fr., syn.: helotium fructigenum (bull.: merat) p. karst., helotiales. localities: trzebnica, wrocław (s c h r o e t e r 1908), wielkopolska province: pniewy, węgrowiec, osowa góra, promno (l i s i e w s k a 1965), łęczna-włodawa lake district, lake długie (c h m i e l 1987), kazimierz landscape park (c h m i e l 1991), roztoczański n. p. (s a ł a t a 1972), bachus reserve (s a ł a t a 1988). host: on fallen nuts and acorns of corylus avellana, fagus sylvatica, quercus and prunus (d e n n i s 1968), carpinus betulus (l i s i e w s k a 1965; c h m i e l 1987, 1991; s a ł a t a 1972, 1988). hymenoscyphus phyllophilus (desm.) kuntze, syn.: helotium phyllophilum (desm.) p. karst., helotiales. locality: anielki near międzyrzec (e i c h l e r 1904). host: on dead branches of fagus (fa r r et al. 1989), carpinus betulus (e i c h l e r l. c.). c h m i e l (unpubl. data) collected this species in poland on leaves of betula, quercus and populus. hypocrea chionea ellis et everh., hypocreales. locality: liski near międzyrzec (e i c h l e r 1907). host: on dead branches of carpinus betulus and salix caprea (e i c h l e r l. c.), pinus (fa r r et al. 1989). the species was noted in europe (e i c h l e r l. c.) and north america (fa r r et al. l. c.). va s i l y e v a (1998) found hypocrea subpachybasioides doi on dead twigs of carpinus cordata in russian far east. microfungi of carpinus betulus 265 hypoxylon atropurpureum (fr.) fr. nemania atropurpurea. hypoxylon fragiforme (pers.) j. kickx. f., xylariales. locality: białowieża n. p. (tr u s z k o w s k a 1965; b u j a k i e w i c z et al. 1997). host: on dead branches and trunks of fagus, corylus, carpinus. it was mostly found on fagus (g r a n m o et al. 1989) and occasionally on other trees. e n d e r l e (1982) noted it on quercus and carpinus betulus. hypoxylon fuscopurpureum (schwein.) m. a. curtis, syn.: hypoxylon vogesiacum (pers.) sacc. var. microsporum j. h. miller, xylariales. locality: białowieża n. p. (c h l e b i c k i et al. 1996). host: on dead wood of carpinus betulus, known also on quercus sp. in north america (fa r r et al. 1989). the species is known from europe and asia on acer pseudoplatanus and quercus sp. (j u , r o g e r s 1996). hypoxylon fuscum (pers.) fr., xylariales. localities: mazurian lake district (tr u s z k o w s k a 1960), białowieża n. p. (tr u s z k o w s k a 1965; c h l e b i c k i et al. 1996), puszcza augustowska forest, czerwone bagno (c h l e b i c k i , unpubl. data). host: on corticated and rarely decorticated branches and stumps of trees, mostly on betulaceae, acer, alnus, betula,carya,castanea, corylus, fagus, fraxinus, ilex, nyssa, ostrya, prunus, quercus, sorbus, tilia (m u n k 1957; d e n n i s 1968; a b e 1986; g r a n m o et al. 1989; fa r r et al. 1989), carpinus cordata (va s i l y e v a 1998). alnus and corylus are recognized as dominant hosts (g r a n m o et al. 1989). the species was noted in northern hemisphere (a b e 1986; g r a n m o et al. l. c.; fa r r et al. l. c.). a very close species hypoxylon macrocarpum pouzar was noted in czech (p o u z a r 1978) and austria (k a h r et al. 1996). hypoxylon howeanum peck, xylariales. localities: międzyrzec (e i c h l e r 1907), białowieża n. p. (c h l e b i c k i et al. 1996), puszcza augustowska forest, czerwone bagno reserve (c h l e b i c k i , unpubl. data), carpathians, beskid niski mts., rzeszówka valley in magura n. p., (c h l e b i c k i unpubl. data). host: on dead, corticated branches and trunks of quercus serrata, q. acutissima, (a b e 1986), acer, betula, castanea, fagus, liriodendron, malus, ostrya, oxydendrum, populus, querus, tilia (fa r r et al. 1989), carpinus cordata and betula mandshurica (va s i l y e v a 1998), acer platanoides, betula pendula, carpinus betulus, corylus avellana, fagus sylvatica, tilia cordata (c h l e b i c k i 1989; b u j a k i e w i c z et al. 1997). corylus, carpinus and fagus were mentioned as dominant hosts in europe (p e t r i n i , m ü l l e r 1986), and quercus in japan. it is common species in poland, noted in europe, asia, japan, north america (d e n n i s 1968; a b e 1986; fa r r et al. 1989; va s i l y e v a l. c.). hypoxylon multiforme (fr.) fr., xylariales. localities: strzelińskie hills (tr u s z k o w s k a , c h l e b i c k i 1983), białowieski n. p. (c h l e b i c k i et al. 1996), ojców nad prądnikiem (wr ó b l e w s k i 1925). host: on dead branches and wood of various frondose trees, especially on betula (m u n k 1957), carpinus betulus (fa k i r o v a 1993), alnus and rarely fagus sylvatica. hypoxylon rubiginosum (pers.) fr., xylariales. localities: strzelińskie hills (tr u s z k o w s k a , c h l e b i c k i 1983), białowieża n. p. (tr u s z k o w s k a 1965; c h l e b i c k i et al. 1996); puszcza romincka forest, 266 a. chlebicki and m. a. chmiel boczki reserve and carpathians, beski niski mts., rzeszówka valley in magura n. p. (c h l e b i c k i , unbupl. data). host: on dead wood and branches of trees, known on 54 host plants (fa r r et al. 1989), noted in poland on corylus avellana, populus tremula, fraxinus excelsior, carpinus betulus and betula pubescens subsp. carpatica (c h l e b i c k i 2002). it is cosmopolitan species (fa r r et al. l. c.). hypoxylon serpens (pers.: fr.) kickx nemania serpens. hypoxylon vogesiacum (pers.) sacc. var. microsporum j. h. miller h. fuscopurpureum. hysterium pulicare pers., hysteriales. locality: białowieża n. p. (c h l e b i c k i et al. 1996). host: on dead wood of betula, cornus, eucalyptus, malus, quercus (fa r r et al. 1989). this species is very common on quercus, betula and alnus in lithuania. kirschsteiniothelia aetiops (berk. et m. a. curtis) d. hawksw., dothideales. locality: białowieża n. p. (c h l e b i c k i et al. 1996). host: on dead wood of populus in north america (fa r r et al. 1989), quercus robur and corylus avellana in sweden (e r i k s s o n 1992) and carpinus betulus in austria (s c h e u e r 1997). the species was noted in temperate regions (fa r r et al. l. c.). laestadia carpinea (fr.) schroet. sphaerognomonia carpinea. ♦ kretzschmaria deusta (hoffm.) p. m. d. martin, xylariales. localities: botanical garden, kraków, skały panieńskie (n a m y s ł o w s k i 1909), białowieża n. p. (c h l e b i c k i et al.. 1996). host: on dead and senescent trunks and stumps of acer, aleurites, alnus, citrus, fagus, ilex, liriodendron, nyssa, poncirus, quercus, tilia, ulmus (fa r r et al. 1989), carpinus betulus and alnus (c h l e b i c k i et al. l. c.). it is cosmopolitan species. lachnum fuscescens var. fuscescens (pers.) p. karst, syn.: dasyscyphus fuscescens (pers.) gray, helotiales. localities: dębina reserve, kampinos n. p. (b o r o w k s a 1966), kazimierz landscape park, parchatka (c h m i e l 1991), białowieża n. p. (c h m i e l 1997), chełm environs., (c h m i e l , unpubl. data). host: noted on fallen leaves of quercus, fagus (d e n n i s 1968) and carpinus betulus, (c h m i e l 1991), roztoczański n. p. on fagus sylvatica (c h m i e l , unpubl. data). lasiosphaeria canescens (pers. ) p. karst., sordariales. locality: puszcza romincka forest, ‘boczki’ reserve (c h l e b i c k i , unpubl. data). host: on decorticated branches of fagus silvatica, quercus (e r i k s s o n 1992) and carpinus betulus. lasiosphaeria hirsuta (fr.) ces. et de not., sordariales. locality: białowieża n. p. (tr u s z k o w s k a 1965). host: on decaying wood of various trees, also carpinus betulus and tilia cordata in białowieża n. p. (tr u s z k o w s k a 1965). lasiosphaeria hispida (tode) fuckel, sordariales. locality: białowieża n. p. (tr u s z k o w s k a 1965). host: on dead twigs of betula pubescens, carpinus betulus (tr u s z k o w s k a l. c.). lasiosphaeria ovina (pers.) ces. et de not., sordariales. microfungi of carpinus betulus 267 locality: białowieża n. p. (tr u s z k o w s k a 1965). host: on decaying wood of various trees, also on betula, carpinus and tilia in białowieża n. p. (tr u s z k o w s k a l. c.). lasiosphaeria spermoides (hoffm.) ces. et de not., sordariales. locality: białowieża n. p. (c h l e b i c k i et al. 1996). host: on dead wood of old stumps, reported mostly on fagus sylvatica, occasionally on other trees (m u n k 1957). lauriomyces catenatus (preuss) r. f. castañeda et w.b. kendr., teleomorph: damatioscypha, helotiales. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead wood of carpinus betulus (b o r o w s k a l. c.). lophiostoma curreyi sacc syn.: lophiostoma hysterioides (schwein.) sacc., dothideales. locality: puszcza romincka forest, żytkiejmska struga reserve (c h l e b i c k i 2005a). host: on decorticated branches of various host plants (fa r r et al. 1989). mamiania fimbriata (pers.: fr.) ces. et de not. gnomonia fimbriata. ♦ massaria carpinicola tul. et c. tul., anamorph: hendersonia carpinicola sacc., pyrenulales. localities: wołów, opole (s c h r o e t e r 1908), białowieża n. p. (tr u s z k o w s k a 1959). massaria urceolata (wallr.) sacc. is also known on carpinus. host: on twigs of carpinus. ♦ melampsoridium carpini (nees) dietel, uredinales. locality: zwierzyniec tenczyński (tęczyński) near chrzanów (r a c i b o r s k i 1888; m a j e w s k i 1977). the species is known in europe, north america, russian far east, china and japan (m a j e w s k i l. c.; z h a n g et al. 1997). host: on leaves of carpinus betulus, c. orientalis, corylus avellana, some asiatic species of carpinus and ostrya, also american species ostrya virginiana (m a j e w s k i l. c.) and carpinus fargesiana (z h a n g et al. 1997). ♦ melanconis chrysostroma (fr. ) tul. et c. tul., syn.: valsa chrysostroma (fr.) tul. et c. tul., diaporthe hyperopta nitschke in g. otth, melanconis hyperopta (nitschke) wehmeyer, melanconis carpinigera (berk. et m. a. curtis) petrak, anamorph: melanconium microsporum nees (e r i k s s o n 1992), diaporthales. localities: puławy (k o c h m a n 1964), upper and lower silesia (tr u s z k o w s k a 1976), strzelińskie hills (tr u s z k o w s k a , c h l e b i c k i 1983), białowieża n. p. (tr u s z k o w s k a 1959, 1965, l. c.; c h l e b i c k i et al. 1996). host: on senescent twigs and branches of hornbeam. the species was noted on carpinus and rarely on quercus, known from europe and north america (we h m e y e r 1941; fa k i r o v a 1993). melanconis chrysostroma var. ellisii (rehm) wehm. is known only from carpinus caroliniana. c o n n e r s (1967) suggested that melanconis chrysostroma var. carpinigera (berk.) wehm. belongs to m. chrysostroma var. ellisii. va s i l y e v a (1987) pointed out that such species as melanconis chrysostroma (fr.) tulasne which inhabits carpinus, melanconis stilbostoma (fr.) tulasne noted on betula, melanconis alni tulasne noted on alnus and melanconis ostryae (dearn.) wehm. noted on ostrya, are very 268 a. chlebicki and m. a. chmiel similar to one another and should be included in one species. also melanconis flavovirens (otth) wehm. on corylus avellana is strictly related to this group of species. ♦ melanconis spodiaea tul et c. tul, syn.: melanconiella spodiaea (tul. et c. tul.) sacc., anamorph: melanconium ramulorum corda (eriksson 1992), diaporthales. localities: niemodlin (s c h r o e t e r 1908). it is uncommon species, reported by m u n k (1957) from danmark, by e r i k s s o n (l. c.) from sweden and noted by d e n n i s (1968) in england. host: on senescent branches of hornbeam. it is peculiar fungus restricted to the carpinus ssp. the ascospores of m. spodiaea are pale brown at full maturity. s a c c a r d o (1882) separated and considered it as a type species of a new genus melanconiella, but we h m e y e r (1941) included it again in melanconis. melanconis xanthostroma (mont.) j. schröt., syn.: wuestneia xanthostroma (mont.) reid et booth, cryptosporella aurea (fuckel) sacc., diaporthe bitorulosa (berk. et broome) sacc., diaporthales. localities: trzebnica, niemcza, strzegom, ziębice, opole (s c h r o e t e r 1908), białowieża n. p. (tr u s z k o w s k a 1959; c h l e b i c k i et al. 1996), noted in europe (r e i d , b o o t h 1989) and usa (fa r r et al. 1989). host: on dead twigs of castanea sativa, carpinus betulus, corylus avellana, ostrya carpinifolia, fagus sylvatica (r e i d , b o o t h l. c.) and liquidambar (fa r r et al. l. c.). melanomma pulvis-pyrius (pers.) fuckel, melanommatales. localities: międzyrzec (e i c h l e r 1907), strzelińskie hills (tr u s z k o w s k a , c h l e b i c k i 1983), białowieża n. p. (tr u s z k o w s k a 1965; c h l e b i c k i et al. 1996); carpathians, beski niski mts., rzeszówka valley in magura n. p. (c h l e b i c k i unpubl. data). host: on dead wood of various plants. it is very common species. melogramma campylosporum fr., syn.: melogramma bulliardi tul. et c. tul., diaporthales. localities: międzyrzec (e i c h l e r 1907), wola justowska near kraków (wr ó b l e w s k i 1925), białowieża n. p. (tr u s z k o w s k a 1959, 1965; c h l e b i c k i et al. 1996). host: on dead branches of carpinus betulus, corylus avellana. va s i l y e v a (1998) described a new species melogramma corylina lar. vass. from russian far east on corylus heterophylla which has somewhat longer spores (45-56 um long). metasphaeria nigrovelata feltg. = sagedia carpinea (pers.) hoehnel, lichenized fungus! monostichella robergei (desm.) hoehn., vide teleomorph: sphaerognomonia carpini. mycosphaerella maculiformis pers.: fr. mycosphaerella punctiformis. menispora ciliata corda, anamorphic fungus, teleomorph: chaetosphaeria, sordariales. localities: kampinos n. p., krzywa góra, zamczysko, tilio-carpinetum, pinocarpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead bark, wood and stems of quercus robur, carpinus betulus, betula pendula, fagus sylvatica, prunus, picea, tilia cordata, calamagrostis, phragmites and tanacetum (b o r o w s k a 1986, l. c.). the fungus is known in europe, canada and new zeland. menispora glauca pers., see chaetosphaeria ovoidea. mollisia ventosa p. karst., helotiales. microfungi of carpinus betulus 269 locality: kruklany in puszcza borecka forest, pino-quercetum and quercocarpinetum communities (g i n k o 1986). host: on dead twigs of pinus sylvestris, salix sp. (s a c c a r d o 1889), carpinus betulus (g i n k o l. c.). the fungus was reported from finland, poland and great britain (s a c c a r d o l. c.; g i n k o l. c.). mycosphaerella punctiformis (pers.) starbäck, mycosphaerellales. localities: liski near międzyrzec (e i c h l e r 1907) lwówek, jawor, ścinawa, syców, opole, henryków, boguszyn near kłodzko (s c h r o e t e r 1908). host: on overwintered leaves, it is plurivorous species, noted on carpinus betulus in bulgaria (fa k i r o v a 1993). ♦ nectria cinnabarina (tode) fr., anamorph: tubercularia vulgaris tode: fr., hypocreales. locality: białowieża n. p. (tr u s z k o w s k a 1959; c h l e b i c k i et al. 1996). host: on living twigs of various plants. it is very common species. nectria coccinea (pers.) fr., hypocreales. locality: białowieża n. p., on dead branch (c h l e b i c k i , unpubl. data). host: on ascomata of various pyrenomycete fungi and wood of trees such as acer, betula, carpinus, castanea, corylus, fagus, fraxinus, hedera, ilex, morus, populus, quercus, sambucus, taxus, tilia, ulmus (b o o t h 1959), ailanthus, albizia, ceanothus, cornus, malus, melia, umbellularia (fa r r et al. 1989). it is cosmopolitan species. nectria episphaeria (tode) fr., syn.: nectria sanguinea (sibth: fr.) fr., hypocreales. localities: białowieża n. p. (tr u s z k o w s k a 1965; c h l e b i c k i , s k i r g i e ł ł o 1995), puszcza augustowska forest (c h l e b i c k i , unpubl. data). host: on old stromata of diatrype stigma, (tr u s z k o w s k a l. c.) as well as on old stromata of diatrype sp., diatrype decorticata and d. stigma (c h l e b i c k i , s k i r g i e ł ł o l. c.). ♦ nectria radicola gerlach et l. nilsson, anamorph: cylindrocarpon destructans (zinss.) scholten, sordariales. locality: wronczyn near poznań (m a ń k a et al. 2002). host: on living roots of carpinus betulus. nemania atropurpurea (fr.) pouzar, syn.: hypoxylon atropurpureum (fr.) fr., xylariales. locality: białowieża n. p. (p o u z a r 1985a). the species is known mainly from temperate regions, taiwan and new guinea (j u , r o g e r s 1999). host: on dead branches and trunks of fagus sylvatica, populus nigra, tilia platyphyllos, ulmus carpinifolia, carpinus betulus, fomes pinicola (p o u z a r l. c.). nemania serpens (pers.) gray, syn.: hypoxylon serpens (pers.) kickx, xylariales. localities: białowieża n. p. (c h l e b i c k i et al. 1996); puszcza romincka forest, boczki reserve (c h l e b i c k i , unpubl. data); the carpathians, beskid niski mts., rzeszówka valley in magura n. p. (c h l e b i c k i , unpubl. data). the species was known in temperate and subtropical regions (fa r r et al. 1989), east china (a b e , l i u 1995). host: on decayinjg wood of various deciduous trees (p o u z a r 1985b), also on carpinus betulus. it was noted in poland on populus tremula, fagus sylvatica, quercus robur (c h l e b i c k i , unpubl. data). ♦ oidium carpini foitzik, teleomorph erysiphe, erysiphales. 270 a. chlebicki and m. a. chmiel localities: kraków, tarnów (p i ą t e k 2004). host: on living leaves of carpinus betulus. it is rare fungus in poland. orbilia leucostigma (fr.) fr., incertae sedis, ascomycetes. locality: kruklany in puszcza borecka forest, pino-quercetum and quercocarpinetum communities (g i n k o 1986). it is a rare species (d e n n i s 1968). host: on dead wood of trees, noted also on carpinus betulus (g i n k o l. c.). ♦ pezicula carpinea (pers.) tul. ex fuckel, anamorph: cryptosporiopsis fasciculata (tode ex tul.) petrak, helotiales. localities: teleomorph: międzyrzec, liski (e i c h l e r 1902), zielona góra, zgorzelec, lwówek śląski, syców, brynica (s c h r o e t e r 1908), warszawa environs, pyry (k o c h m a n 1971), las piwnicki reserve (h o ł o w n i a 1977); anamorph: białowieża n. p., (c h l e b i c k i , unpubl. data). host: on twigs of carpinus betulus, c. caroliniana, fagus, castanea and populus (d e n n i s 1968; c o n n e r s 1967; tr u s z k o w s k a , c h l e b i c k i 1983). anamorph was earlier reported from strzeliñskie hills (tr u s z k o w s k a , c h l e b i c k i l. c.) on populus tremula. g r z y w a c z (1993) reported it from poland as dermatea carpinea (pers.) rehm. c o n n e r s (l. c.) reported anamorph on carpinus caroliniana from ontario (canada). phaeostalagmus cyclosporus (grove) w. gams, anamorphic fungus, incertae sedis, ascomycetes. locality: kampinos n. p. (b o r o w s k a 1987). host: on dead bark and wood of acer, alnus, betula, castanea, fagus, fraxinus, picea, prunus, ulmus, quercus, corylus, ilex, rhododendron, sambucus, ulex, carex, rumex (b o r o w s k a 1986). phaeostalagmus tenuissimus (corda) w. gams et hol.-jeh., anamorphic fungus, incertae sedis, ascomycetes. locality: białowieża n. p., kampinos n. p. (b o r o w s k a 1987). host: on rotten wood and fallen fruits of castanea, fagus, ilex, picea, pinus, quercus, rubus, salix, carpinus betulus. the fungus was reported in europe, canada and kazakhstan (b o r o w s k a 1986). ♦ phyllactinia guttata (wallr.) lév., erysiphales. localities: miradz near strzelno, lipno, zalesie near piaseczno, górzez near jawor, wałbrzych, szczytów near dzierżoniów, bochotnica naer kazimierz dolny, włostowice near puławy (s c h r o e t e r 1908; s a ł a t a 1985). host: on living leaves, plurivorous species. it occurs in poland on acer negundo, alnus glutinosa, a. incana, betula obscura, b. pendula, b. pubescens subsp. pubeacens and subsp. carpatica, corylus avellana, c. sanguinea, c. tubulosa, crataegus oxyacantha, fagus sylvatica, fraxinus excelsior, quercus robur, syringa vulgaris and ulmus minor (h e n n i n g s 1892; s c h r o e t e r l. c., s a ł a t a l. c.). this species is rare in lithuania on betulaceae (g r i g a l i û n a i t ë 1997). besides it occurs on 76 species of host plants in usa (fa r r r et al. 1989). ♦ phyllosticta carpini schulzer et sacc., anamoorphic fungus, teleomorph: guignardia, dothideales. localities: siedlce (tr z e b i ń s k i et al. 1916), malta near poznań, as phyllosticta carpinea sacc. (d o m i n i k 1936). the fungus was reported from europe and north america (fa r r et al. 1989). host: on living leaves of carpinus betulus, alnus (tr z e b i ń s k i et al. l. c.; fa r r et al. l. c.). microfungi of carpinus betulus 271 pleurophragmium rousselianum (mont.) s. hughes pseudospiropes rousselianus. polydesmia pruinosa (gerd ex berk. et broome) boud., helotiales. locality: kruklanki in puszcza borecka forest, circaeo-alnetum (g i n k o 1986). it is very common fungus (d e n n i s 1968). host: on dead twigs and branches of various trees, also on carpinus betulus (g i n k o l. c.), dead twigs and ascocarps as well as on ostioles of perithecia of other fungi (d e n n i s l. c.). propolis farinosa (pers.) fr., syn.: propolomyces farinosus (pers.) sherwood, propolomyces versicolor (fr.: fr.) dennis, propolis faginea (schrad.) p. karst., rhytismatales. localities: czerwieńsk near zielona góra (s c h r o e t e r 1908). host: mostly on dead wood of various trees, noted in great britain on acer, chamaenerion, corylus, fagus, fraxinus, ilex, larix, lonicera, malus, pinus, populus, prunus, quercus, rhamnus, salix, sambucus, sorbus and tilia (d e n n i s 1968). it is a very common species, noted also on branches, pine cones, woody stems (d e n n i s l. c.), on subfossil oak quercus robur and cutted by beavers trunks (c h l e b i c k i unpubl. data). pseudospiropes rousselianus (mont.) m. b. ellis, syn.: pleurophragmium rousselianum (mont.) s. hughes, anamorphic fungus, incertae sedis, ascomycetes. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead wood of carpinus betulus (b o r o w s k a l. c.). rhinocladiella atrovirens nannf., anamorphic fungus, teleomorph: capronia, chaetothyriales. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead wood of carpinus betulus (b o r o w s k a l. c.). rosellinia corticium (schw.) sacc, xylariales. locality: białowieża n. p. (c h l e b i c k i et al. 1996; c h l e b i c k i 2006). host: on dead bark and wood of various trees (m u n k 1957). it is one of the most common species of rosellinia in europe. according to p e t r i n i (1993) many collections identified as rosellinia aquila (fr.) de not., syn.: r. byssiseda (tode) schroeter, in fact are not this fungus. thus it is also possible that r. corticium was earlier included in r. aquila. host: on dead bark and wood of various trees, on acer pseudoplatanus, eucalyptus sp., ilex aquifolium, salix sp., salvia officinalis and vitis vinifera. fa k i r o v a (1993) noted it in bulgaria on carpinus betulus, va s i l y e v a (1998) noted the same fungus on branches of carpinus cordata in russian far east. rosellinia mammiformis (pers.) ces. et de not., xylariales. locality: białowieża n. p. (c h l e b i c k i et al. 1996). host: known on dead wood, stems and branches of various herbs and trees. rutstroemia bolaris (batsch.) rehm, helotiales. localities: międzyrzec, liski (e i c h l e r 1902), wrocław (s c h r o e t e r 1908), białowieża n. p. (s k i r g i e ł ł o 1960), roztoczański n. p. (s a ł a t a 1972), wołwinów near chełm (s a ł a t a , b e d n a r c z y k 1977), kazimierz landscape park (c h m i e l 1991). host: on fallen twigs of carpinus betulus, c. caroliniana, corylus and casta272 a. chlebicki and m. a. chmiel nea (s c h r o e t e r l. c.; c o n n e r s 1967; fa r r et al. 1989). it is known from north america and europe. ♦ sclerotinia sclerotiorum (lib.) de bary, helotiales. localities: wronczyn near poznań (mańka et al. 2002). host: in living roots of carpinus betulus. sphaerognomonia carpinea (fr.) potiebnia apiosporopsis carpinea. ♦ splanchnonema carpini (fuckel) m. e. barr, syn.: pleomassaria carpini (fuckel) sacc., anamorph: hendersonia carpini sacc., pleosporales. localities: zielona góra, wołów, ziębice, opole (s c h r o e t e r 1908), bielany near kraków (wr ó b l e w s k i 1918), białowieża n. p. (tr u s z k o w s k a 1965). host: on senescent twigs of carpinus betulus, carpinus caroliniana (fa r r et al 1989; b a r r 1990). the species was noted in europe and north america (fa r r et al. l. c.). stilbospora macrosperma pers. ex mérat teleomorph: hapalocystis bicaudata. taeniolella exilis (p. karst.) s. hughes, anamorphic fungus, teleomorph: glyphium, hysteriales. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead wood and bark of betula (e l l i s 1971). the species is know from europe and north america (e l l i s l. c.). taeniolina scripta (p. karst.) m. e. kirk, syn.: taeniolella scripta (p. karst.) s. hughes, anamorphic fungus, teleomorph: glyphium, hysteriales. localities: białowieża n. p. (c h l e b i c k i unpubl. data). host: on dead wood of betula, corylus, fagus and sorbus aucuparia (e l l i s 1971). the species is known from europe. ♦ taphrina carpini (rostr.) johansson, taphrinales. localities: s a ł a t a (1975) cited many localities of this fungus gathered by m a g n u s (1895), s c h r o e t e r (1908), s z u l c z e w s k i (1938), r o u p p e r t (1912), z a l e s k i and g l a s e r (1953), tr z e b i ń s k i (1916). it has been noted in strzelińskie hills (s c h r o e t e r 1908), pomorze lake district, wielkopolska and małopolska upland, the sudetes and the west carpathians (s a ł a t a 1974). host: it is witches’ brooms on twigs and branches noted in europe on carpinus betulus and c. orientalis (s a ł a t a 1974) whereas in north america taphrina australis (atk.) giesenh. occurs on carpinus caroliniana (c o n n e r s 1967; fa r r et al. 1989). a related species, taphrina virginica sadebeck is known from ostrya in north america (fa r r et al. 1989). moreover taphrina coerulescens (desmaz. et mont) tul., leaf blister occasionally was noted on ostrya (fa r r et al. l. c.). trematosphaeria pertusa (pers.) fuckel, pleosporales. locality: białowieża n. p. (tr u s z k o w s k a 1965). host: on decorticated branches of carpinus betulus, corylus avellana, populus tremulus, tilia cordata (tr u s z k o w s k a l. c.). trimmatostroma betulinum (corda) s. hughes, anamorphic fungus, incertae sedis, ascomycetes. locality: kampinos n. p., krzywa góra, tilio-carpinetum (b o r o w s k a 1987). the species is known from europe (e l l i s 1971). host: on dead twigs of betula. e l l i s (l. c.); b o r o w s k a (l. c.) noted this fungus on betula pendula, quercus robur and carpinus betulus. microfungi of carpinus betulus 273 tubeufia cerea (berk. et m. a. curtis) höhn, helicosporium vegetum nees, anamorphic fungus, pleosporales. localities: kampinos n. p., krzywa góra, sieraków, zamczysko, tilio-carpinetum, pino-carpinetum, pino-quercetum (b o r o w s k a 1987). host: on dead wood of carpinus betulus, quercus robur, tilia cordata (b o r o w s k a l. c.). ♦ valsa ambiens (pers.) fr., anamorph: cytospora leucosperma (fr.) fuckel, diaporthales. localities: czerwieńsk (zielona góra region), wrocław (s c h r o e t e r 1908), białowieża n. p. (c h l e b i c k i et al. 1996). host: on senescent twigs, noted on over 40 species in north america (fa r r et al. 1989). wuestneia xanthostroma (mont.) reid. et booth melanconis xanthostroma. xylaria corniformis (fr.) fr., xylariales. localities: białowieża n. p. (l e s s ø e 1987; c h l e b i c k i et al. 1996), puszcza romincka forest, boczki reserve (c h l e b i c k i , unpubl. data). host: on branches and decorticated wood of alnus, fagus, olea, populus (tr a v e r s o 1906), carpinus, fagus, quercus (fa r r et al. 1989) fagus, carpinus betulus (l a e s s ø e 1987). it was noted in poland on alnus glutinosa and carpinus betulus (c h l e b i c k i , unpubl. data). the species was reported from europe, north africa, asia and north america (l a e s s ø e l. c.; fa r r et al. l. c.). xylaria hypoxylon (l.) grev., xylariales. localities: dolina szwajcarka valley near ciechocinek (r o u p p e r t 1909), białowieża n. p. (c h l e b i c k i et al. 1996). host: on dead branches and stumps, it is plurivorous species known also on carpinus betulus. xylaria longipes nitschke, xylariales. locality: białowieża n. p. (c h l e b i c k i et al. 1996). host: on dead stumps and branches, mostly reported on acer. zignoëlla fallax (sacc.) sacc., sordariales. locality: białowieża n. p. 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(in:) z. m i r e k (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków, 812 pp. w r ó b l e w s k i a. 1918. przyczynek do znajomości grzybów galicji zachodniej. spraw. kom. fizjogr. 52: 122 127. w r ó b l e w s k i a. 1925. champignons recueillis par m. raciborski dans les environs de cracovie et dans le tatra en 1883 et 1890. acta soc. bot. pol. 3: 29 41. z a l e s k i k., g l a s e r t. 1953. parasitic and saprophytic fungi (exept agaricaceae a. boletaceae) of state forests wolsztyn (province poznań, poland) collected in 1949 1950. acta soc. bot. pol. 22: 633 652. z h a n g n i n g , z h u a n g y i o u y u n , we i s h u x i a 1997. fungal flora of the dabe mountains: uredinales. mycotaxon 61: 49 79. grzyby mikroskopijne grabu carpinus betulus w polsce s t r e s z c z e n i e praca zawiera krytyczną listę grzybów mikroskopijnych zebranych na grabie w polsce. ogółem odnotowano 115 taksonów, w tym 28 gatunków pasożytniczych. najliczniejszym ga tunkiem okazał się grzyb gnomonia fimbriata, odnotowano także bardzo rzadkie gatunki jak camarops plana, c. polysperma, lasiosphaeria hirsuta, lophiostoma curreyi, nemania atropur purea, oidium carpini i inne. stwierdzono także obecność 10 gatunków wyłącznych dla grabu (wyspecjalizowanych troficznie): camarops plana, diaporthe carpini, encoelia carpini, gno monia fimbriata, massaria carpinicola, melampsoridium carpini, melanconis spodiaea, oidium carpini, pezicula carpinea i phyllosticta carpini. 2014-01-01t11:44:31+0100 polish botanical society 2014-08-31t21:57:25+0200 polish botanical society 2014-09-03t19:47:44+0200 polish botanical society 2014-09-02t20:10:23+0200 polish botanical society 2014-09-06t20:23:54+0200 polish botanical society 2014-09-06t20:22:19+0200 polish botanical society 2014-09-08t11:10:58+0200 polish botanical society 2014-09-03t19:46:27+0200 polish botanical society 2014-09-03t19:47:54+0200 polish botanical society a contribution to the lichen biota of belarus vladimir v. golubkov1 and martin kukwa2 1department of botany, ya kupala hrodna state university azheshka 22, by 230023 hrodna, vgolubkov@grsu.by 2department of plant taxonomy and nature protection, university of gdańsk al. legionów 9, pl 80 441 gdańsk, dokmak@univ.gda.pl g o l u b k o v v.v., k u k w a m.: a contribution to the lichen biota of belarus. acta mycol. 42 (1): 155 164, 2006. details of 11 lichen and 1 lichenicolous fungus taxa occurring in belarus are presented, of which 6 lichens (cliostomum leprosum, fellhanera gyrophorica, lecanora thysanophora, lepraria eburnea, l.jackii, l.neglecta) and 1 lichenicolous fungus (tremella hypogymniae) are newly reported for the country; new localities of recently reported lepraria elobata and l. incana s.str. are presented and the occurrence of chrysothrix candelaris, loxospora elatina and pertusaria coronata is confirmed. the taxonomy of chrysothrix candelaris is briefly discussed and localities of cliostomum leprosum from poland are reported. key words: sorediate lichens, lichenicolous fungus, new species, eastern europe introduction the lichen biota in central and east-central europe is reasonably well studied; however, in many regions some groups of species are still very poorly known, particularly the sorediate crustose lichens and lichenicolous fungi which have been neglected in the past. other groups have also been under-recorded, as well as many taxa not recognized until recently, even though some were common constituents of the biota. this paper contributes to our knowledge of the belarussian lichen biota by reporting some new or noteworthy taxa belonging mainly to the commonly or constantly sterile sorediate lichens, as well as the lichenicolous fungus, tremella hypogymniae. material and methods the lichen collection of v.v. golubkov has been studied, including chemical analyses using thin layer chromatography (tlc) according to the methods proposed by orange et al. (2001). for each taxon, notes are provided on their characteristics acta mycologica vol. 41 (1): 155-164 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 156 v. v. golubkov and m. kukwa and similar species are discussed; general distribution details are provided for taxa new to belarus. the lichenicolous fungus is marked with an asterisk (*). list of species chrysothrix candelaris (l.) j.r. laundon this species is rather widespread in europe, but its chemistry has only been studied in some countries (l a u n d o n 1981; tø n s b e r g 1992), and in central and central-eastern europe the chemical variability has never been studied. our specimens contain pinastric acid and according to tø n s b e r g (1992) belong to chemotype 1. however, k a l b (2001), who revised the concept of c. candelaris, recognized tropical entities with pinastric acid as a separate species, c. xanthina (vain.) kalb. this taxon had been considered to be only a synonym of c. candelaris (laundon 1981), but kalb found, in addition to chemistry, some differences in anatomy, distribution and ecology, and pointed out that the pinastric acid containing samples of c. candelaris s.l. from temperate regions may represent an as yet undescribed taxon. as there are no differences between specimens containing calycin (c. candelaris s.str.) and pinastric acid (suggested undescribed species) in temperate regions, we propose here to include them in c. candelaris, at least temporarily. however, this problem needs further studies based on molecular analyses. specimens examined: belarus, brestskaya oblast’, kamenetskiï region, belovezhskaya pushcha (forest), forest section no 741, oak forest, on trunk of oak, 20.07.1973, leg. d.i. tret’yakov; brestskaya oblast’, pruzhanskiï region, belovezhskaya pushcha, forest section no 589, vicinity of pererevo, oak forest with hornbeam near river, on trunk of maple, 22.06.1983, leg. v.v. golubkov. cliostomum leprosum (räsänen) holien & tønsberg this species is new to the country. it is a rare lichen taxon in europe and has been reported from finland (s a n t e s s o n et al. 2004), norway, sweden (tø n s b e r g 1992; s a n t e s s o n et al. 2004), poland (fa ł t y n o w i c z 2003) and switzerland (d i e t r i c h , s c h e i d e g g e r 1996). details of polish localities have never been published until know (see additional specimens examined). c. leprosum is characterised by the sorediate whitish-grey thallus producing atranorin and caperatic acid, very rarely accompanied by an unidentified fatty acid (tø n s b e r g 1992). all specimens from belarus and poland contained atranorin and caperatic acid, but the additional fatty acid was detected in specimen from ‘mechacz’ nature reserve. the species can be mistaken for some ochrolechia species with a whitish thallus and not delimited soralia, especially o. microstictoides räsänen. however only c. leprosum produces atranorin with caperatic acid (tø n s b e r g 1992). specimens examined: belarus, minskaya oblast’, myadel’skiï region, exact locality unknown, old railway, pine forest, on bark of pine, 14.06.1984, leg. v.v. golubkov. additional specimens examined: poland, romincka forest, n part of the ‘mechacz’ nature reserve, atpol grid square af85, on bark of pine, 17.05.1985, leg. s. cieśliński & z. tobolewski (ktc, duplicate in bg and ugda-l); bielska plain, białowieża forest, ‘michnówka’ nature reserve, forest section no 572; atpol grid square cg64, on bark of pine in peat bog pine forest, 11.08.2002, leg. p. czarnota a contribution to the lichen biota of belarus 157 (gpn 2984, duplicate in ugda-l); augustowska plain, augustowska forest, wigierski national park, forest section no 159n; atpol grid square bf29, on bark of pine in peat bog pine forest, 08.11.1997, leg. w. fałtynowicz (wrsl, duplicate in ugda-l 9329). fellhanera gyrophorica sérus., coppins, diederich & scheid. this species is new to the country. only recently it has been described from austria, lithuania, luxembourg, poland, switzerland and ukraine (s é r u s i a u x et al. 2001). additionally it has been found in estonia (m o t i e j ū n a i t ė , p r i g o d i n a l u k o š i e n e 2002) and lithuania (m o t i e j ū n a i t ė et al. 2003), and further localities were discovered in poland (c z y ż e w s k a et al. 2001, 2002, 2005; s p a r r i u s 2002). when described, f. gyrophorica was known only from sterile collections with pycnidia (s é r u s i a u x et al. 2001). apothecia were discovered in the same year in material collected in poland (s p a r r i u s 2001) and fertile material has also been reported from lithuania (m o t i e j ū n a i t ė et al. 2003). the specimen from belarus possesses only pycnidia. f. gyrophorica is usually sterile and easily recognisable by its rather large brown pycnidia reacting c+ red and pyriform conidia. some bacidia s.lat. and micarea species can look similar, but they can be easily separated by the pycnidial characters (s é r u s i a u x et al. 2001). specimen examined: belarus, brestskaya oblast’, pruzhanskiï region, 3 km w of babinec village, belovezhskaya pushcha, forest section no 560a, on bark of black alder, 26.07.1983, leg. v.v. golubkov. lecanora thysanophora r.c. harris this species is new to belarus. it was described from north america (h a r r i s et al. 2000), and than reported from many european countries (tø n s b e r g 1999; p r i n t z e n et al. 2002; k o w a l e w s k a , k u k w a 2003; m o t i e j ū n a i t ė et al. 2003; g u t t o v á , palice 2004; m r a k et al. 2004; c z y ż e w s k a et al. 2005; k u k w a 2005). l. thysanophora is a sorediate lichen producing a characteristic fibrillose prothallus which is well developed in at least some part(s) of the thallus. it always contains atranorin, usnic acid, zeorin and terpenoids called ‘thysanophora unknowns’ (h a r r i s et al. 2000). some north american specimens also produce porphyrillic acid (h a r r i s et al. 2000); however, no porphyrillic acid has been detected in european specimens to date (k u k w a , unpubl.). the species can be very easily mistaken for haematomma ochrolecum (neck.) j.r. laundon; however, they can be separated by their chemistry since l. thysanophora produces ‘thysanophora unknowns’ which are not present in h. ochrolecum (h a r r i s et al. 1999). when the fibrillose prothallus is not well developed, l. thysanophora can remind one of l. expallens ach., but l. thysanophora contains atranorin and ‘thysanophora unknowns’, whereas l. expallens contains xanthones in addition to usnic acid and zeorin (tø n s b e r g 1992; h a r r i s et al. 1999). specimens examined: belarus, gomel’ckaya oblast’, mozyrskiï region, vicinity of proftyuki village, oak forest, on oak, 26.08.1977, leg. v.v. golubkov; gomel’ckaya oblast’, mozyrskiï region, vicinity of simonovichskiï mlynok village, pripyatskiï national park, spruce forest, on bark of black alder, 16.08.1982, leg. v.v. golubkov; grodnenskaya oblast’, novogrudskiï region, forest section no. 56, oak forest with 158 v. v. golubkov and m. kukwa spruce and hornbeam, on bark of hornbeam, 24.06.1987, leg. v.v. golubkov; grodnenskaya oblast’, svislochskiï region, belovezhskaya pushcha, forest section no 82, lime tree forest, on bark of lime tree, 17.06.1983, leg. v.v. golubkov; brestskaya oblast’, pruzhanskiï region, 3 km w of babinec village, belovezhskaya pushcha, forest section no 560a, on black alder, 26.07.1983, leg. v.v. golubkov. lepraria eburnea j.r. laundon this species is new to belarus. it is rather widespread in europe (e.g. laundon 1992; tø n s b e r g 1992; orange 1997; k u k w a 2002a; b a y e r o v á , k u k w a 2003), and rarely reported in north america (l a u n d o n 1992; o r a n g e 1997), asia (g ü v e n ç , o z t ü r k 1999; c o b a n o g l u , a k d e m i r 2000) and oceania (new zealand) (w i r t h 1997). l. eburnea is well characterised by its leprose thallus consisting of soredia producing some projecting hyphae and the presence of alectorialic acid (usually barbatic acid is also detectable). commonly alectorialic acid is accompanied by protocetraric acid (chemotype i), very rarely with psoromic acid (chemotype iii; known only from great britain), but also quite commonly it occurs alone (chemotype ii) (o r a n g e 1997; see also l a u n d o n 1992 and tø n s b e r g 1992). sometimes it is very difficult to determine the chemotype, since the additional substances may occur in very low concentrations. the belarusian specimen probably belongs to chemotype i, as the habitat is more suitable for it (see k u k w a 2002a); however, further studies with more sensitive methods should be carried out to solve its identity. chemically, l. eburnea can be mistaken only for l. neglecta (nyl.) erichsen, as both produce alectorialic acid. however, the morphology and habitat requirements are different: l. eburnea has a leprose thallus and occurs on rather shaded habitats protected from direct rain, whereas l. neglecta has a granular course thallus and grows in places exposed to sun and rain (e.g. l a u n d o n 1992; tø n s b e r g 1992; kukwa 2002a). some species, such as l. incana (l.) ach. and l. lobificans nyl., have a similar morphology, but they do not produce alectorialic acid (cf. l a u n d o n 1992; tønsberg 1992). specimen examined: belarus, brestskaya oblast’, kamenetskiï region, vicinity of kamenyuki village, belovezhskaya pushcha, forest section no 778, oak forest, on decaying oak, 17.07.1983, leg. v.v. golubkov. lepraria elobata tønsberg this species has been only recently reported from belarus by c z y ż e w s k a & k u k w a (2005). although known from only four collections, it is probably a common species, as it is in neighbouring poland (k u k w a 2002a, b). the species is characterised by the predominantly grey thallus, the absence of a medulla, soredia with a well developed wall and lacking projecting hyphae, and the presence of atranorin, zeorin and stictic acid complex (tø n s b e r g 1992). l. lobificans is identical in its chemistry, but differs by its usually greenish woolly thallus and airy soredia, in many cases with highly visible long projecting hyphae (tø n s b e r g 1992). l. elobata can be mistaken for l. incana, but it is easily separated by the chemistry, as it produces divaricatic acid and zeorin (tø n s b e r g 1992). specimens examined: belarus, minskaya oblast’, minskiï region, 1.5 km n slabodshchina village, on bark of pine, 25.04.1976, leg. g.n. antonov & o.m. maclovskiï; minskaya oblast’, minskiï region, 2.3 km n of slabodshchina village, on bark of pine, a contribution to the lichen biota of belarus 159 25.04.1976, leg. o.m. maclovskiï; vitebskaya oblast’, berezinckiï nature reserve, 0.1 km ne of ‘gurba’ range, on bark of oak, 16.08.1973, leg. luk’yanov. lepraria incana (l.) ach. the first belarusian record was recently confirmed by tlc (c z y ż e w s k a , k u k w a 2005). the species is probably very common, as it is, for example, in neighbouring poland (k u k w a 2002a). l. incana is well characterized by the production of divaricatic acid and zeorin (e.g. tø n s b e r g 1992). the only other european taxon with divaricatic acid and zeorin, l. crassissima (hue) lettau, differs in its production of large amounts of nordivaricatic acid, which is only present in traces in l. incana (b o o m et al. 1994). specimens examined: belarus, grodnenskaya oblast’, novogrudskiï region, 2 km e of vcelyubci, on mossy decaying log, 23.08.1981, leg. o.m. maclovskiï; brestskaya oblast’, belovezhskaya pushcha, forest section no 824, on bark of black alder, 16.06.1983, leg. v.v. golubkov; brestskaya oblast’, belovezhskaya pushcha, forest section no 98, spruce forest, on bark of spruce, 16.06.1983, leg. v.v. golubkov; brestskaya oblast’, belovezhskaya pushcha, forest section no 824, peat bog pine forest, on decaying wood, 10.08.1986, leg. v.v. golubkov; brestskaya oblast’, kamenetskiï region, vicinity of kamenyuki village, belovezhskaya pushcha, forest section 829, oak forest with hornbeam, on oak, 17.07.1983, leg. v.v. golubkov; brestskaya oblast’, kamenetskiï region, 5 km e of pashuki village, belovezhskaya pushcha, forest section no 890b, oak forest, in the crevices of tree bark 28.07.1983, leg. v.v. golubkov; grodnenskaya oblast’, svislochskiï region, belovezhskaya pushcha, forest section no 82, lime tree forest, on decaying spruce log, 17.08.1982, leg. v.v. golubkov; gomel’ckaya oblast’, mozyrskiï region, vicinity of cumonovichckiï mlynok village, pripyatskiï national park, spruce forest, on bark of spruce, 15.08.1982, leg. v.v. golubkov. lepraria jackii tønsberg this species is new to belarus. it is rather widespread in europe (e.g. k ü m m e r l i n g et al. 1995; k u k w a 2002a; b a y e r o v á , k u k w a 2003), and is also reported from asia, australia and north america (k ü m m e r l i n g et al. 1995). l. jackii prefers bark of coniferous trees, and according to k ü m m e r l i n g et al. (1995) it has widened its distribution due to the preference of such trees in forest management. it also commonly occurs on birches, and can be more widely distributed because of the popularity of planting such trees (kukwa, unpubl.). l. jackii is characterised by the powdery, green-grey thallus containing atranorin, jackinic (very rarely absent) and roccellic (usually present) acids (tø n s b e r g 1992; l e u c k e r t et al. 1995). the most similar chemistry is known from l. borealis lohtander & tønsberg, but this species has a granular thallus and prefers exposed to rain habitats, whereas l. jackii has a powdery thallus and grows on substrata sheltered from direct rain (e.g. l o h t a n d e r 1994). l. rigidula (de lesd.) tønsberg and l. toensbergiana bayerová & kukwa also produce atranorin and fatty acids, but they also produce nephrosteranic and toensbergianic acids, respectively (l e u c k e r t et al. 1995; b a y e r o v á et al. 2005) specimens examined: brestskaya oblast’, kamenetskiï region, vicinity of kamenyuki village, belovezhskaya pushcha, forest section no 582/601, on bark of pine, 08.08.1983, leg. v.v. golubkov; grodnenskaya oblast’, novogrudskiï region, vicinity of novogrudok village, on bark of spruce, 06.06.1989, leg. v.v. golubkov; minskaya 160 v. v. golubkov and m. kukwa oblast’, minskiï region, 2 km n of slabodshchina village, pine forest, on bark of pine, 03.10.1976, leg. g.n. antonov & o.m. maclovskiï; vitebskaya oblast’, polotskiï region, near lake glubokoe, pine forest, on deacying wood, 20.07.1985, leg. v.v. golubkov. lepraria neglecta (nyl.) erichsen this species is new to belarus. it is a cosmopolitan lichen known from europe (e.g. l a u n d o n 1992; tø n s b e r g 1992; k ü m m e r l i n g et al. 1993; m o t i e j ū n a i t ė 1999; k u k w a 2002a), asia (k ü m m e r l i n g et al. 1993; aptroot and seaward 1999), australia (kümmerling et al. 1993), north america (k ü m m e r l i n g et al. 1993) and antarctica (o l e c h 2001). morphologically l. neglecta resembles other species of the so-called l. neglecta group: l. borealis lohtander & tønsberg, l. cacuminum (a. massal.) lohtander and l. caesioalba (de lesd.) j.r. laundon. however, it can be very easily distinguished by the chemistry, as it is the only species producing alectorialic acid as a major compound (e.g. tø n s b e r g 1992; l o h t a n d e r 1994; l e u c k e r t et al 1995). chemically l. neglecta is very similar to l. eburnea, but the morphology easily separates these two taxa (for differences, see under that species). specimen examined: belarus, vitebskaya oblast’, glubokskiï region, border of lake dolgoe, on stone, 11.05.1985, leg. v.v. golubkov. loxospora elatina (ach.) a. massal. this species has been reported from belarus by to m i n (1956) and g o r b a c h (1973), but these identifications were based only on spot test reactions. the present record is the first one confirmed by tlc. l. elatina is characterized by its greyish areolate thallus with at least at the beginning discrete soralia. it produces thamnolic acid and ± elatinic acid (s c h r e i n e r and h a f e l l n e r 1992; tø n s b e r g 1992). it can be mistaken for some ochrolechia species, but it differs in its chemical constituents. specimen examined: belarus, vitebskaya oblast’, verkhodvinskiï region, no detailed locality, on birch, 20.06.1986, leg. v.v. golubkov. pertusaria coronata (ach.) th. fr. this species has been reported from belarus by to m i n (1956), g o r b a c h (1973) and g o l u b k o v (1987), but the record below is the first one confirmed by tlc. there are only two other isidiate pertusaria species which can be mistaken for p. coronata, namely p. coccodes (ach.) nyl. and p. flavida (dc.) j.r. laundon. however, these species have very distinctive chemistry. p. coronata produces coronaton and stictic acid complex which gives a p+ orange reaction of the thallus, p. coccodes has norstictic acid which gives a k+ yellow changing to red reaction, and p. flavida posses xanthones, which gives a c+ orange reaction (cf. tø n s b e r g 1992). specimen examined: belarus, grodnenskaya oblast’, svislochskiï region, belovezhskaya pushcha, forest section no 82, lime tree forest, on lime tree, 17.06.1983, leg. v.v. golubkov. *tremella hypogymniae diederich & m.s. christ. this lichenicolous fungus is new to belarus. it is rather widespread in europe (d i e d e r i c h 1996; c z y ż e w s k a et al. 2001, 2002; k u k w a et al. 2002; c z y ż e w s k a 2003; k u k w a 2004), and also known from north america (d i e d e r i c h 1996). a contribution to the lichen biota of belarus 161 this species, belonging to the heterobasidiomycetes, induces the formation of convex, pale to pinkish, sometimes dark or black, galls on the thalli of hypogymnia physodes (l.) nyl. (d i e d e r i c h 1996). specimen examined: belarus, grodnenskaya oblast’, svislochskiï region, belovezhskaya pushcha, forest section no 82, lime tree forest, on thallus of hypogymnia physodes growing on lime tree, 17.06.1983, leg. v.v. golubkov. acknowledgements. we are very grateful to professor krystyna czyżewska (university of łódź) for her valuable help during the preparation of the paper and professor mark r.d. seaward (university of brad ford) for checking the english. we would like to thank also professor stanisław cieśliński (świętokrzyska academy), dr. paweł czarnota (gorce national park) and professor wiesław fałtynowicz (university of wrocław) for making available their specimens of cliostomum leprosum. references a p t r o o t a . , s e a w a r d m . r . d . 1999. annotated checklist of hongkong lichens. trop. bryology 17: 57 101. b a y e r o v á š . , k u k w a m. 2003. new records of leprarioid lichens in the czech republic. biologia, bratislava 59: 19 23. b a y e r o v á š . , k u k w a m . , f e h r e r j. 2005. a new species of lepraria (lichenized ascomycetes) from europe. bryologist 108: 131 138. b o o m p. p. g . v a n d e n , b r a n d m . , d i e d e r i c h p. , a p t r o o t a . , s é r u s i a u x e. 1994. re port of lichenological field meeting in luxembourg. bull. socc. nat. luxemb. 95: 145 176. c o b a n o g l u g . , a k d e m i r b . 2000. new records for turkish lichen flora. in: the fourth ial sym posium progress and problems in lichenology at the turn of the millenium, barcelona, 3 8 septem ber 2000. book of abstracts, p. 92. c z y ż e w s k a k . 2003. distribution of some lichenicolous fungi in poland. acta mycol. 38 (1/2): 111 122. c z y ż e w s k a k . , c i e ś l i ń s k i s . , m o t i e j ū n a i t ė j . , k o l a n k o k . 2002. the budzisk nature reserve as a biocentre of lichen diversity in knyszyńska large forest (ne poland). acta mycol. 37 (1/2): 77 92. c z y ż e w s k a k . , k u k w a m . 2005. notes on two species of lepraria from belarus. graphis scripta 17: 20 21. c z y ż e w s k a k., m o t i e j ū n a i t ė j., c i e ś l i ń s k i s. 2001. species of lichenized and allied fungi new to bialowieża large forest (ne poland). acta mycol. 36 (1): 13 19. c z y ż e w s k a k . , m o t i e j ū n a i t ė j . , c i e ś l i ń s k i s . 2005. new and noteworthy species of lichens and allied fungi from north eastern poland. acta mycol. 40 (2): 277 291. d i e d e r i c h p. 1996. the lichenicolous heterobasidiomycetes. biblioth. lichenol. 61: 1 198. d i e t r i c h m . , s c h e i d e g g e r c . 1996. the importance of sorediate crustose lichens in the epiphytic lichen flora of the swiss plateau and the pre alps. lichenologist 28: 245 256. f a ł t y n o w i c z w. 2003. the lichens, lichenicolous and allied fungi of poland an annotated checklist. 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[lichens of muránska planina national park iii cigánka]. reussia 1, suppl. 1: 11 47. g ü v e n ç ş . , o z t ü r k ş . 1999. lichens in the north east regions of cyprus. feddes repert. 110: 455 463. h a r r i s r . c . , b r o d o i . m . , tø n s b e r g t. 2000 (2001). lecanora thysanophora, a common leprose lichen in north america. bryologist 103: 790 793. k a l b k . 2001. new or otherwise interesting lichens. i. biblioth. lichenol. 78: 141 167. 162 v. v. golubkov and m. kukwa k o w a l e w s k a a . , k u k w a m . 2003. additions to the polish lichen flora. graphis scripta 14: 11 17. k u k w a m . 2002a. lepraria ach. and leproloma cromb. in poland. biblioth. lichenol. 82: 67 76. k u k w a m . 2002b. porosty z rodzajów lepraria i leproloma w puszczy białowieskiej. parki nar. rez. przyr. 21: 253 262. k u k w a m . 2004. new or interesting records of lichenicolous fungi from poland ii. species mainly from northern poland. herzogia 17: 67 75. k u k w a m . 2005. lecanora thysanophora (lecanoraceae, zlichenizowane ascomycota) w polsce. fragm. flor. geobot. polonica 12: 385 391. k u k w a m . , m o t i e j ū n a i t ė , j . , r u t k o w s k i , p. , z a l e w s k a , a . 2002. new or interesting records of lichenicolous fungi from poland i. herzogia 15: 129 139. k ü m m e r l i n g h . , l e u c k e r t c . , w i r t h v. 1991. chemische flechtenanalysen vi. lepraria in cana (l.) ach. nova hedwigia 53: 507 517. k ü m m e r l i n g h . , tr i e b e l d . , r a m b o l d g . 1993. lepraria neglecta and its lichenicolous fungi. bilbioth. lichenol. 53: 147 160. k ü m m e r l i n g h . , l e u c k e r t c . , w i r t h v. 1995. chemische flechtenanalysen xi. lepraria jackii tønsberg. nova hedwigia 60: 457 465. l a u n d o n j . r . 1981. the species of chrysothrix. lichenologist 13: 101 121. l a u n d o n j . r . 1992. lepraria in the british isles. lichenologist 24: 315 350. l e u c k e r t c . , k ü m m e r l i n g h . , w i r t h v. 1995. chemotaxonomy of lepraria ach. and leprolo ma nyl ex. crombie, with particular reference to central europe. biblioth. lichenol. 58: 245 259. l o h t a n d e r k . 1994. the genus lepraria in finland. ann. bot. fenn. 31: 223 231. m o t i e j ū n a i t ė j . 1999. checklist of lichens and allied fungi of lithuania. botanica lithuanica 5: 251 269. m o t i e j ū n a i t ė j . , p r i g o d i n a l u k o s i e n e i . 2002. chaenothecopsis rubescens new to lithuania and fellhanera gyrophorica new to estonia. graphis scripta 13: 43 44. m o t i e j ū n a i t ė j . , k u k w a m . , c z a r n o t a p. , p r i g o d i n a l u k o š i e n ė i . , h i m e l b r a n t d . , k u z n e t s o v a e . , k o w a l e w s k a a . 2003. lichens and allied fungi collected during the 15th symposium of baltic mycologists and lichenologists in birštonas, lithuania. botanica lithu anica 9: 109 119. m r a k t. , m a y r h o f e r h . , b a t i č f. 2004. contributions to the lichen flora of slovenia xi. lichens from the vicinity of lake bohinj (julian alps). herzogia 17: 107 127. o l e c h m . 2001. annotated checklist of antarctic lichens and lichenicolous fungi. institute of botany of jagiellonian university, kraków, 145 pp. o r a n g e a . 1997. chemical variation in lepraria eburnea. lichenologist 29: 9 13. p r i n t z e n c . , h a l d a j . , p a l i c e z . , tø n s b e r g t. 2002. new and interesting lichen records from old growth forest stands in the german national park bayerischer wald. nova hedwigia 74: 25 49. s a n t e s s o n r . , m o b e r g r . , n o r d i n a . , tø n s b e r g t. , v i t i k a i n e n o . 2004. lichen form ing and lichenicolous fungi of fennoscandia. museum of evolution, uppsala university, uppsala, 359 pp. s c h r e i n e r e . , h a f e l l n e r j . 1992. sorediöse, corticole krustenflechten im ostalpenrum. i. die flechtenstoffe und die gesicherte verbreitung der besser bekannten arten. biblioth. lichenol. 45: 1 291. s é r u s i a u x e . , c o p p i n s b . j . , d i e d e r i c h p. , s c h e i d e g g e r c . 2001. fellhanera gyrophorica, a new europaean species with conspicuous pycnidia. lichenologist 33: 285 289. s p a r r i u s l . b . 2002. discovery of apothecia confirms generic position of fellhanera gyrophorica. li chenologist 34: 86. to m i n m . p. 1956. opredelitel’ korkovych lishaïnikov evropeïskoï chasti sssr. izdatel’stvo akademii nauk belaruskoї ssr, minsk, 532 pp. tø n s b e r g t. 1992. the sorediate and isidiate, corticolous, crustose lichens in norway. sommerfeltia 14: 1 331. tø n s b e r g t. 1999. lichenes isidiosi et sorediosi crustacei exsiccati. schedae to fascicle 2 (nos 26 50): 1 10. bergen, norway. tø n s b e r g t. , t ü r k r . , h o f m a n n p. 2001. notes on the lichen flora of tyrol (austria). nova hedwigia 72: 487 497. a contribution to the lichen biota of belarus 163 w i r t h v. 1997. additional lichen records from new zealand 21. candelariella coralliza, lepraria ebur nea, racodium rupestre, rinodina olivaceobrunnea, rinodina pyrina and trapeliopsis flexuosa. aus tralasian lichenological newsletter 40: 11 13. materiały do bioty porostów białorusi s t r e s z c z e n i e biota porostów europy środkowej i środkowo wschodniej jest stosunkowo dobrze po znana, choć pewne jej grupy są ciągle słabo zbadane. dotyczy to zwłaszcza sorediowanych, zwykle płonnych porostów skorupiastych oraz grzybów naporostowych. w wyniku tego wiele gatunków częstych, a nawet pospolitych w innych obszarach europy, nie zostało dotychczas odnotowanych w poszczególnych krajach tej części kontynentu. celem niniejszej pracy jest uzupełnienie danych o porostach sorediowanych białorusi. spośród 12. prezentowanych taksonów, sześć gatunków porostów (cliostomum leprosum, fel lhanera gyrophorica, lecanora thysanophora, lepraria eburnea, l. jackii, l. neglecta) oraz je den grzyb naporostowy (tremella hypogymniae) zostały podane po raz pierwszy dla białorusi. ponadto wykazano nowe stanowiska dla niedawno odnotowanych lepraria elobata i l. incana, potwierdzonych za pomocą chromatografii cienkowarstwowej (por. c z y ż e w s k a i k u k w a 2005). gatunki te, jak i pozostali przedstawiciele rodzaju lepraria są najprawdopodobniej bardzo częstymi składnikami ekosystemów na terenie białorusi. ponadto potwierdzono za pomocą analizy wtórnych metabolitów porostowych występowanie chrysothrix candelaris, lo xospora elatina i pertusaria coronata. okazy ch. candelaris reprezentują chemotyp 1 (stwier dzono kwas pinastrowy). według niektórych badaczy (por. k a l b 2001) ta rasa chemiczna może reprezentować gatunek jeszcze nieopisany dla nauki; wymaga to jednak dalszych badań, w tym także molekularnych. dla cliostomum leprosum podano także stanowiska z polski, któ re nie były nigdy wcześniej publikowane, pomimo odnotowania tego taksonu w kraju (por. f a ł t y n o w i c z 2003). 2014-01-01t11:44:03+0100 polish botanical society 2014-09-03t19:47:17+0200 polish botanical society 2014-09-02t20:10:50+0200 polish botanical society 2014-09-05t16:23:33+0200 polish botanical society 2014-09-05t16:23:54+0200 polish botanical society © the author(s) 2014 published by polish botanical society grazing preference of ceratophysella sp. 1 (collembola) for pseudoperonospora cubensis – fungal pathogen of cucumis sativus a. chlebicki1 and j.m. radwański2 1department of mycology, w. szafer institute of botany, polish academy of sciences lubicz 46, pl-31-512 kraków, a.chlebicki@botany.pl 2institute of systematics and evolution of animals, polish academy of science sławkowska 17, pl-31-016 kraków, jmr1@o2.pl chlebicki a., radwański j.m.: grazing preference of ceratophysella sp. 1 (collembola) for pseudoperonospora cubensis – fungal pathogen of cucumis sativus. acta mycol. 49 (1): 93–97, 2014. grazing preferences of collembolans ceratophysella sp. 1 were noted. the collembolans feed on the sporangia of pseudoperonospora cubensis growing on plants cucumis sativus in greenhouses in esfahan province in iran. sporangia of pseudoperonospora cubensis were the preferred food of all fungal species noted in the alimental tract of the springtails. all digestive tracts of the springtails contained sporangia of p. cubensis. key words: cucurbit downy mildew, pseudoperonospora cubensis, springtails, feeding preferences introduction there are many interactions between insects, fungi and plants, i.e. botanophila, epichloë and dactylis/puccinellia (górzyńska et al. 2010, 2011) or phalacrus, anthracoidea and carex (steiner 1984; chlebicki 2007, 2011). many microarthropods are mycophagous. some species of collembolans feed on the mycelium of pathogenic fungi (sabatini, innocenti, 2000, lootsma, scholte, 1997). also grazing preferences of collembolans for endomycorrhizal fungi are well known (timm, larink 1995). there is single article devoted collembolan species sinella curviseta which feeding of mycelium of fusarium oxysporum f. sp. cucumerinum parasitizing cucumber seedlings (nakamura et al. 1992). we found the next collembolan species feeding on plant pathogen pseudoperonospora cubensis (berk. & m.a. curtis) rostovzev growing on leaves of cucumis sativus. the collected springtails belong to the unknown species of the genus ceratophysella from c. denticulata group that might be a new species to the world (nematollahi et al. 2009). acta mycologica vol. 49 (1): 93–97 2014 doi: 10.5586/am.2014.009 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 94 a. chlebicki and j.m. radwański © the author(s) 2014 published by polish botanical society material and methods collembolans were found feeding on cucumber seedlings in greenhouses in esfahan province during 2005-2006. juvenile individuals was preserved on 96% ethanol. then, they were prepared as fallow: 18 hours in lactophenol (lactic acid 100 ml, glycerol 200 ml, distilled water 100 ml, phenol 100 g), 10-40 minutes in 10% koh, few seconds of heating in lactophenol. at the end of the specimen was closed on a slide in the medium marc andré ii (distilled water 50 ml, chloral hydrate 200 gram, glycerol 30 gram, gum arabic 20 gram). 70 specimens of collembolans were checked up for fungal sporangia. 30 sporangia of p. cubensis were measured. the morphological characters of the fungi and collembolan body were examined using light microscopy (nikon smz 1500, nikon eclipse 80i). microphotographs were taken with these microscopes equipped with a digital camera nikon ds r1. morphological terminology of the mildew was used after savory et al. (2011). results and discussion pseudoperonospora cubensis (berk. & m.a. curtis) rostovzev, annals inst. agron. moscow 9: 47, 1903. sporangia elliptical, lemon-shaped, grey-purple 22-27(33) x 17-25 µm. sporangiophores slender, branches in the dichotomous manner, branches and tips straight, tips gradually tapering to a point (fig. 1a). all digestive tracts of the springtails contain sporangia of p. cubensis (fig. 2). moreover spores of other species were found. only three dark spores of acremonium group (gliomastix, pseudogliomastix) together with spores of p. cubensis were found (fig. 1b). the spores (fig. 1b) were similar to these of plate 52 a (seifert et al. 2011). in the same figure a single ornamented spore is presented. in the next picture (fig. 1c) are showed pollen grains of unidentified plant. some hyphae of dematiaceous fungus are illustrated in the figure 1d. the cucurbit downy mildew (pseudoperonospora cubensis) is currently the most destructive disease of cucumber which have been responsible for annual yield losses of up to 80% (lebeda, urban 2007; savory et al. 2011). it has wide host range and occurs approximately on 20 different genera of cucurbitaceae (lebeda, urban 2007). sporangia of pseudoperonospora cubensis became the preferred food of all fungal species noted in the alimental tract of the springtails. sporangia lying in alimentary tract of collembolans are mostly disfigured. it suggest that ceratophysella sp. 1 not disperse sporangia of the fungus via gut passage. nematollahi et al. (2009) suggest that in the infested plants secondary roots were devoured completely by collembolans and the plants could be easily infected by pathogens. but our observations indicate that collembolans reduced number of fungal sporangia. however, the fungal sporangia can be transported by the springtails on their body and this way contributing to the spread of the disease. amount of sporangia transported on springtail bodies is distinctly smaller than its amount in alimental tract. on the basis of our investigation it is difficult to precise determination of influence of the collembolan species on cucumber cultivation in greenhouse. ceratophysella sp. 1 is probably a rare species with occurrence grazing preference of ceratophysella for pseudoperonospora cubensis 95 © the author(s) 2014 published by polish botanical society fig. 1. a – sporangia and sporangiospores of pseudoperonospora cubensis, b – 3 dark spores of acremonium group fungus and single hyaline, ornamented spore, c – plant grains, d –dematiaceous hyphae. fig. 2. springtail ceratophysella sp. 1 juv. with spores in the alimental track. 96 a. chlebicki and j.m. radwański © the author(s) 2014 published by polish botanical society restricted to the near east. other species of the c. denticulata group have similar food preference (babenko et al. 1994). next investigation are necessary for recognition of influence of ceratophysella sp. 1 on pathogenic fungi of greenhouse plants. it is possible that these collembolans can be used for protection against pathogenic fungi (biological control agent – bca) as in the case of proisotoma minuta (isotomidae) used for suppression of rhizoctonia solani on cotton in a greenhouse environment (lartey et al. 1994, 2008). acknowledgements. we thank dr. marcin piątek for suggestions and two reviewers for comments. financial support was provided by the w. szafer institute of botany of the polish academy of sciences through its statutory fund. references babenko a.b., chernova n.m., potapov m.b., stebaeva s.k. 1994. opredelitel kollembol fauny rossii i soprezhelnykh stran: semeistvo hypogastruridae. moskva, nauka. chlebicki a. 2007. notes on the distribution and ecology of fungi of the genus anthracoidea (ustilaginomycetes) in poland. polish bot. j. 52 (2): 151-158. chlebicki a. 2011. anthropogenic origin of the smut fungus anthracoidea caricis population in the gorce mts (poland). polish bot. j. 56 (2): 333-337. górzyńska k., lembicz m., olszanowski z., leuchtmann a. 2010. an unusual botanophila-epichloë association in a population of orchardgrass (dactylis glomerata) in poland. journal of natural history 44: 2817-2824. górzyńska k., lembicz m., olszanowski z., leuchtmann a. 2011. botanophila-epichloë interaction in a wild grass, puccinellia distans lacks dependence on the fly vector. annals of the entomological society of america 104 (4): 841-846. lartey r.t., curl e.a., peterson c.m. 1994. interactions of mycophagous collembolan and biological control fungi in the suppression of rhizoctonia solani. soil biology and biochemistry 26(1):81-88. http://dx.doi.org/10.1016%2f0038-0717%2894%2990198-8 lartey r.t., curl e.a., peterson c.m., williams j.l. 2008. control of rhizoctonia solani and cotton seedling disease by laetisaria nivalis and mycophagous insect proisotoma minuta (collembola). journal of phytopatology 133 (2): 89-98. lebeda a., urban j. 2007. temporal changes in pathogenicity and fungicide resistance in pseudoperonospora cubensis populations. acta horticulturae 731: 327-336. lootsma m., scholte k. 1997. effect of soil moisture content on the suppression of rhizoctonia stem canker on potato by the nematode aphelenchus avenae and the springtail folsomia fimetaria. plant pathology 46 (2): 209-215. http://dx.doi.org/10.1046%2fj.1365-3059.1997.d01-229.x nakamura y., matsuzaki i., itakura j. 1992. effect of grazing by sinella curviseta (collembola) on fusarium oxysporum f. sp. cucumerinum causing cucumber disease. pedobiologia 36 (3): 168-171. nematollahi m., bagheri m., radwanski j.m. 2009. new reports of collembola for iran with surveying of the importance in the greenhouses of esfahan province, iran. plant protection journal 1 (3): 327-335. sabatini m.a., innocenti g. 2000. soil-borne plant pathogenic fungi in relation to some collembolan species under laboratory conditions. mycological research 104 (10): 1197-1201. http://dx.doi. org/10.1017%2fs0953756200003026 savory e.a., granke l.l., quesada-ocampo l.m., varbanova m., hausbeck m.k., day b. 2011. the cucurbit downy mildew pathogen pseudoperonospora cubensis. molecular plant pathology 12 (3): 217-226. http://dx.doi.org/10.1111%2fj.1364-3703.2010.00670.x seifert k. morgan-jones g., gams w., kendrick b. 2011. the genera of hyphomycetes. cbs-knaw fungal biodiversity centre, utrecht. steiner w.e. 1984. a review of the biology of phalacrid beetles (coleoptera). (in): q. wheeler and blackwell m. (eds). fungus-insect relationships. new york: columbia university press: 424-445. timm t., larink o. 1995. grazing preferences of some collembolan for endomycorrhizal fungi. biology and fertility of soils 19: 266-268. http://dx.doi.org/10.1007%2fbf00336171 grazing preference of ceratophysella for pseudoperonospora cubensis 97 © the author(s) 2014 published by polish botanical society preferencje pokarmowe skoczogonka z rodzaju ceratophysella do grzyba pseudoperonospora cubensis – grzybowego patogena ogórków cucumis sativus streszczenie preferencje pokarmowe skoczogonków z rodzaju ceratophysella sp. 1 (nowy gatunek do opisania) zostały przedstawione. skoczogonki spożywały sporangia grzyba pseudoperonospora cubensis, który zaatakował rośliny cucumis sativus rosnące w szklarni w prowincji esfahan w iranie. sporangia grzyba pseudoperonospora cubensis były notowane w układzie pokarmowym wszystkich badanych skoczogonków. 2014-06-30t22:45:21+0200 polish botanical society 2014-09-05t16:23:13+0200 polish botanical society 2014-09-03t19:46:10+0200 polish botanical society 2014-09-05t16:24:25+0200 polish botanical society 2014-09-02t20:09:53+0200 polish botanical society 2014-11-20t23:11:10+0000 polish botanical society 2014-08-31t21:56:07+0200 polish botanical society 2014-09-06t20:25:24+0200 polish botanical society 2014-09-02t20:09:12+0200 polish botanical society 2014-08-31t21:57:28+0200 polish botanical society observations on the mycorrhizal status of polygonum viviparum in the polish tatra mts. (western carpathians) michał ronikier1 and piotr mleczko2 1w. szafer institute of botany, polish academy of sciences, lubicz 46, pl 31 512 kraków, michal.ronikier@ib pan.krakow.pl 2institute of botany, jagiellonian university, lubicz 46, pl 31 512 kraków, ubmleczk@cyf kr.edu.pl r o n i k i e r m . , m l e c z k o p. : observations on the mycorrhizal status of polygonum viviparum in the polish tatra mts. (western carpathians). acta mycol. 41 (2): 209 222, 2006. polygonum viviparum is one of very few herbaceous plants known to form ectomycorrhiza; in the tatra mts. it is one of dominants in the alpine zone, but also descends down to the feet of the massif. specimens of this plant were collected from 5 sites at the altitude range 900 2150 m, from granite and limestone. it allowed an estimation of the ectomycorrhizal diversity as well as preliminary ecological observations. roots were also stained in order to check potential presence of arbuscular mycorrhizal colonization. ectomycorrhizae were present in all specimens (with 2 5 morphotypes observed on single plants). in total, 17 morphotypes were observed and briefly described. the most widespread were the mycorrhiza of cenococcum geophilum and a brightly coloured morphotype resembling the ectomycorrhizae of russula sp. no important differences in ectomycorrhizal colonization between low and high localities were found. observed general differences in abundance and diversity of mycorrhiza in p. viviparum between sites could most probably be connected with plant community composition (presence/absence of ectomycorrhizal shrubs maintaining ectomycorrhizal fungi), although mycorrhizae were present also in sites devoid of other ectomycorrhizal plants. structures associated to arbuscular colonization (vesicles, hyphal coils) were occassionally observed, but without formation of arbuscules. key words: polygonum viviparum, ectomycorrhiza, arbuscular mycorrhiza, arctic alpine ecology, cenococcum geophilum, rhizosphere introduction polygonum viviparum l. (polygonaceae) is a widely distributed arctic-alpine species (p a w ł o w s k a 1972), commonly found in the arctic tundra and high mountain regions of the northern hemisphere. in the tatra mts. (the highest massif of the carpathians, with maximum altitude of 2663 m) it is one of dominant species in the alpine and subnival zones (up to 2480 m a.s.l. in the lodowy massif; p a w ł o w s k i acta mycologica vol. 41 (2): 209-222 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 210 m. ronikier and p. mleczko 1956), but also descends down to the meadows at feet of the massif (900 m), which makes a very large altitudinal range. it is one of very few herbaceous plants known to form ectomycorrhiza (reviewed by g a r d e s and d a h l b e r g 1996). h e s s e l m a n (1900) first reported the presence of ectomycorrhizal tips with a hartig net on the root system of p. viviparum. this observation was then confirmed for plants from different sites as central alps (c o n s t a n t i n and m a g r o u 1926; p e y r o n e l 1930, 1937; f o n t a n a 1977), eastern alps (h a s e l w a n d t e r and r e a d 1980; r e a d and h a s e l w a n d t e r 1981; b l a s c h k e 1991a, 1991b), tatra mts. (d o m i n i k et al. 1954), rocky mts. (l e s i c a , a n t i b u s 1986; m a s s i c o t t e et al. 1998), alaska (tr e u et al. 1996). however, some authors reported lack of ectomycorrhizae in their observations in both arctic (b l e d s o e et al. 1990; v ä r e et al. 1992) and mountain (n e s p i a k 1953) sites, which suggests the existence of some constraints in the formation of symbiotic relationship of this type. most literature data are limited to the statement of the presence/absence of ectomycorrhizal structures, while there is only limited information on the diversity of p. viviparum ectomycorrhiza. f o n t a n a (1977) gave account of the diversity of ectomycorrhizae of p. viviparum from italian alps, reporting presence of 16 morphotypes. tr e u et al. (1996) mentioned different ectomycorrhizal tips morphology. m a s s i c o t t e et al. (1998) described anatomical features of two herbaceous plants’ ectomycorrhizae: p. viviparum and kobresia myosuroides. some data suggest presence of two types of mycorrhizal colonization in p. viviparum. the paralell colonization of root system by arbuscular mycorrhizal fungi and ectomycorrhizal fungi was reported by s t a h l (1900) from arctic site and by b l a s c h k e (1991a, 1991b) from the bavarian alps. n e s p i a k (1953) also mentioned the amf colonization of roots of p. viviparum in the tatra mts. the aim of the present work was to describe the mycorrhizal status of p. viviparum in the polish tatra mts. (western carpathians) and to contribute to the study of the diversity of mycorrhiza of this interesting arctic-alpine species. it is thought as a pilot study initiating more complex analyses of ectomycorrhizae in alpine habitats and their role in building the arctic-alpine macromycete diversity in the tatra mts. materials and methods description of the study sites and sampling of plant specimens. five sampling sites were chosen on the stations of polygonum viviparum growing on granitic and calcareous bedrock in polish tatra mts., in different parts of the altitudinal range of the species (tab. 1). samples of plant root systems were collected twice during the vegetation period on three sites and once on two others (tab. 1); 4–5 plants were collected from each site. plants were taken together with the embedding soil (about 20×20×15 cm), transported in plastic bags and stored in a fridge (when quickly analyzed) or frozen at –20° c. samples were soaked in water, then roots of p. viviparum were washed and carefully separated from roots of other plants. only roots connected to rhizome were considered. ectomycorrhizal colonization was analyzed and afterwards the roots were stained for checking the endophytic colonization. em observations – light and scanning electron microscopy. ectomycorrhizal tips were observed submerged in water, using dissecting microscope. morphotypes were distinguished according to methods described by a g e r e r (1986, 1987–2002, 1991), observations on the mycorrhizal status 211 using mantle “scrapings” for plan views. in some cases (abundant morphotypes) basic macrochemical stainings (koh, feso4, sulphovanilin, melzer’s reagent) were also made. non-identified morphotypes were named following rules proposed by a g e r e r (1996). additionally, ectomycorrhizal structure on longitudinal and cross sections were observed. ectomycorrhizal tips for sections were embedded in the synthetic resin (historesin embedding kit – leica, germany; prepared according to manufacturer’s instructions) and cut on the microtome (leica rm 2135, germany; 6−7 μm). slides were observed using microscope with differential interference contrast (dic). ectomycorrhizal tips were stored in faa solution (g e r l a c h 1972). for observations in scanning electron microscope, fresh ectomycorrhizal tips were fixed in 2% glutaraldehyde solution in cacodylate buffer and dehydrated in the increasing acetone and ethanol concentration series (m a s s i c o t t e et al. 1987). em quantitative comparison. quantitative comparison of ectomycorrhizal colonization was very difficult to estimate due to the tight mixture of roots in case of alpine grassland samples and thus only approximate data could be obtained. ectomycorrhizal tips were counted and related to the length of the roots, as follows: m= number of mycorrhizal tips length of roots [cm] each site was characterized by a mean m value from all specimens (4–5). additionally, the ratio of alive and dead mycorrhizal tips (ml/d) was calculated. am colonization. for estimation of endophytic colonization, roots of p. viviparum were stained according to the modified method of p h i l i p s and h a y m a n (1970). roots were softened using 7% koh solution, washed with water and bleached with h2o2 containing nh3 (10:1 v/v) for a few minutes. the material was then acidified in 5% lactic acid solution and stained with 0,01% cotton blue (anilin blue, methyl blue) ta b l e 1 description of sampling sites (all within the western carpathians, the tatra mts.). in the “sampling” column: 1 sampling in may 1997; 2 sampling in september 1997 no site location sampling other em plants in the vicinitydescription alt. [m] coordinates 1 n slopes of the nosal peak (1205 m); fresh meadow at forest edge 910 n 49°16’55” e 19°59’10” 12 picea abies 2 sw slopes of the przełęcz między kopami pass (1550 m); clearings among dwarf pine shrubs 1545 n 49°15’07” e 20°00’13” 12 pinus mugo 3 sw slopes of the kasprowy wierch peak (1985 m); alpine grassland on granite 1960 n 49°13’56” e 19°58’50” 12 [distant stands of salix herbacea] 4 w crest of the szpiglasowy wierch peak (2172 m); alpine grassland on granite 2150 n 49°11’53” e 20°02’28” 2 5 n slopes of the małołączniak peak (2069 m); alpine grassland on limestone 2070 n 49°14’12” e 19°55’15” 2 dryas octopetala, salix reticulata 212 m. ronikier and p. mleczko solution in lactic acid. all steps were conducted in room temperature and lasted (apart from bleaching) 24 h each. stained material was stored in pure lactic acid. prior to staining the roots were kept in 50% ethanol solution. results diversity of ectomycorrhizae on polygonum viviparum. seventeen morphotypes of ectomycorrhizae were isolated from collected samples (cf. fig. 1a–d). the highest number of morphotypes was found on the sites 1, 5 and 2 (10, 8 and 7 morphotypes, respectively) (tab. 2). the highest morphotype number per sample (plant) was 5–6 on three mentioned sites, whereas on the sites 3 and 4 it did not exceed 2. one morphotype was identified as formed by cenococcum geophilum fr. based on presence of clamps, six morphotypes were identified as members of basidiomycota (“p. lanata”, “p. vulpina”, “p. aurata”, “p. tuberoidea”, “p. aspera”, “p. tenua”). ectomycorrhizae were formed mainly on delicate, secondary roots, the side branches of dark roots growing from the rhizome. all ectomycorrhizae were simple and did not form any ramifications. in some tips traces of renewed growth were visible in the form of slight segmentation (beaded mycorrhiza; a g e r e r 1991). morphotypes differed significantly in tips length. some were short (ca. 1 mm), and often of big diameter (e.g. “polygonirhiza epidermoidea”, “p. lacteocinerea”), while some others were elongated (up to 3 mm). a high diversity of fungal mantle structures was also observed. several morphotypes had a primitive, plectenchymatous mantle (e.g. “p. vulpina”), or the mantle of an intermediate character between plectenchymatous and pseudoparenchymatous, with hyphae distinguishable yet considerably thickened (e.g. “p. maculata”, “p. salebrosa” and “p. fusca”). pseudoparenchymatous mantles of different types were represented by “polygonirhiza epidermoidea”, “p. lacteocinerea”, “p. aurata” and “p. tuberoidea”. morphotypes varied also in respect of extramatrical structures. some of them produced very abundant extramatrical mycelium, forming ta b l e 2 synopsis table of the morphotype occurrence in sampling sites (numbers of sites refer to tab. 1) site morphotype 1 2 3 4 5 cenococcum geophilum ■ ■ ■ ■ ■ “p. arenaria” ■ “p. aspera” ■ ■ “p. aurata” ■ “p. epidermoidea” ■ “p. fusca” ■ ■ ■ “p. granulosa” ■ “p. lacteocinerea” ■ ■ ■ ■ ■ “p. lanata” ■ ■ “p. maculata” ■ “p. radiata” ■ “p. rufocystidiata” ■ “p. salebrosa” ■ “p. tenua” ■ “p. terrea” ■ “p. tuberoidea” ■ “p. vulpina” ■ ■ ■ observations on the mycorrhizal status 213 even woolly clusters, as in “p. lanata”, only few emanating hyphae were produced by e.g. “p. terrea” and “p. vulpina”, a smooth mantle surface was observed e.g. in “p. epidermoidea”. abundant, very fine emanating hyphae of “p. arenaria” were covered with a secretion causing sticking of sand particles. cystidia were present in three morphotypes: “p. aurata”, “p. tuberoidea” and “p. rufocystidiata”. hartig net was of paraepidermal type in all examined ectomycorrhizae (a g e r e r 1991), fungal colonization was limited to anticlinal walls of rhizodermal cells (fig. 1f). the hyphae of hartig net were ramified and closely cohering together forming “palmetti” structures. root cells of hartig net zone were strongly elongated transversally (ccq 0.32, comp. a g e r e r 1987-2002). key for the determination of ectomycorrhizae. the dichotomous key is presented below in order to facilitate the identification of morphotypes described in this paper. both macroscopical features (colour, surface structure) and microscopical characteristics of fungal mantle and extramatrical structures in “plan view” were included. 1. mycorrhiza brownish-black or black ....................................................................... 2 1*. mycorrhiza of other colour .................................................................................... 7 2. surface rough and shiny, with long, rigid and dark emanating hyphae. star-like mantle structure. mantle homogenously black (fig. 1a), thick and rigid, densely plectenchymatous, outer layer formed by thick-walled, strongly pigmented cells, forming distinctive star-like arrangements (fig. 1e) (in their centers the “cells” are small, centripetally elongated). emanating hyphae thick-walled, with regularly distributed septa. clamps lacking. inner mantle layer formed by hyphae with weaker stained, thinner walls. mycorrhiza rarely > 1 mm long, usually of a big diameter. .................................................................................................... cenococcum geophilum 2*. mycorrhiza with other features .............................................................................. 3 3. abundant orange-brown cystidia present, well visible under dissecting microscope. mycorrhiza very dark brown, blackish, mantle surface slightly rough (fig. 1b). cystidia elongated, thick-walled with small diameter, narrowing to a sharp top, without clamps at base (fig. 2g). outer mantle layer pseudoparenchymatous, cells rounded, of different sizes. deeper layer dense, intermediate between plectenchymatous and pseudoparenchymatous ............................ “polygonirhiza rufocystidiata” 3*. mycorrhiza without orange-brown cystidia .............................................................. 4 4. mantle (plectenchymatous or pseudoparenchymatous) with visible star-like structure (but never with long, rigid emanating hyphae; cf. 2 − cenococcum geophilum) ........... 5 4*. mantle pseudoparenchymatous, epidermoid or angular, without star-like pattern ... 6 5. mantle pseudoparenchymatous, built by angular cells. thick (> 5 μm in diameter), dark brown emanating hyphae with grainy surface and clamps. mycorrhiza very dark brown with a rigid, cloddish surface. outer mantle layer built of big, strongly stained polygonal cells forming a characteristic „rosette” pattern (fig. 2b) (cells dimensions diminish centripetally). inner layer pseudoparenchymatous, cells thiner-walled and without pattern. emanating hyphae frequently ramified and interconnected. intrahyphal hyphae present .......................................................... “polygonirhiza aspera” 214 m. ronikier and p. mleczko 5*. mantle plectenchymatous. extramatrical structures not observed. mycorrhiza brownish-black. mantle surface slightly rough. outer mantle layer a dense plectenchyma built by long, irregular hyphae converging radially and forming a „rosette” pattern. inner layer pseudoparenchymatous, formed by round cells. ..................................................................................................... “polygonirhiza radiata” 6. outer mantle pseudoparenchymatous with angular hyphal cells. inner mantle layer intermediate between plectenchyma and pseudoparenchyma. emanating hyphae fine, hyalin without clamps, with rare, fine septa and slightly incrusted cell walls. mycorrhiza black, surface slightly cloddish. ....................... “polygonirhiza salebrosa” 6*. outer mantle pseudoparenchymatous with epidermoid hyphal cells. inner mantle intermediate between plectenchymatous and pseudoparenchymatous. towards the inside of the mantle hyphae become thiner and less stained. extramatrical structures not observed mycorrhiza dark brown to blackish. mantle surface very rough and slightly shiny. ........................................................................................................ “polygonirhiza fusca” 7. light or dark brown coloured mycorrhiza .............................................................. 8 7*. mycorrhiza golden-greenish, orange-brown or orange ..................................... 11 7**. mycorrhiza whitish, grey, yellowish or pinkish ................................................. 13 8. light-brown coloured mycorrhiza with distinctly rough, cloddish mantle surface. mantle thick and rigid, plectenchymatous, hyphae brown and thick-walled, often triangular in shape, usually arranged in a distinct „rosette-like” arrangment (fig. 2c); responsible for the cloddish macroscopic mantle character. extramatrical structures not observed .......................................................................... “polygonirhiza granulosa” 8*. mycorrhiza with other features .............................................................................. 9 9. mycorrhiza brown. mantle thin, intermediate between plectenchymatous and pseudoparenchymatous, formed by thin-walled hyphae without clamps. emanating hyphae rare, hyaline, ramified, with clamps and opened anastomoses. ........................................................................................................ “polygonirhiza tenua” 9*. mycorrhiza unhomogeneously brown (with irregular, darker spots). mantle surface smooth and shiny ................................................................................................. 10 10. mycorrhiza dark brown with greyish shade, with distinct darker spots. mantle surface smooth and shiny. mantle thin, plectenchymatous. in the top part of mycorrhizal tip fine, thin-walled emanating hyphae without clamps ........................................................................................................ “polygonirhiza terrea” 10*. light brown mycorrhiza with slight olivaceous shade and distinct small darker spots (fig. 1d). mantle surface smooth, shiny. mantle intermediate between plectenchymatous and pseudoparenchymatous (fig. 2d), hyphae immersed in matrix material. extramatrical structures not observed ................ “polygonirhiza maculata” 11. mycorrhiza orange-brown. outer mantle layer plectenchymatous, formed by a dense net of hyphae, with characteristic groups of paralell hyphae. inner layer formed by larger hyphae, closer to irregular pseudoparenchyma. emanating hyphae rare, fine, hyalin, thin-walled, with clamps. cystidia lacking. mycelium without colour reaction to koh, feso4 and sulphovanilin ..................................................................................................... “polygonirhiza vulpina” observations on the mycorrhizal status 215 11*. mycorrhiza orange or goldenish, with pseudoparenchymatous mantle. numerous hyalin, elongated cystidia, often with clamps ..................................................... 12 12. mycorrhiza short spiny, yellowish-brown with golden-green glaze (fig. 1c). mantle surface smooth and shiny, with hyalin cystidia. outer mantle layer a very loose hyphal net, growing on intermediate, pseudoparenchymatous layer with angular, thin-walled cells. inner mantle layer formed by small, irregular cells. cystidia growing from angular cells, short and obtuse (top rounded), most with a clamp in 1/3 or 1/2 of their lenght. cystidium wall much thicker in the proximal part and thinner above the clamp .......................................................................... “polygonirhiza aurata” 12*. mycorrhiza short spiny, orange. mantle surface smooth or slightly fibrous with numerous hyaline cystidia. outer mantle layer a loose net of thick hyphae without clamps, with rare anastomoses. intermediate layer pseudoparenchymatous with angular, thick-walled cells; the structure of inner layers less regular. cystidia very numerous, obtuse, thick-walled, often with clamps at septa (fig. 1f). this morphotype resembles macroscopically those formed by tuber sp. (although such identity is excluded by presence of clamps). although macroscopically very different from “p. aurata” , microscopical structure and cystidia resemble that morphotype. it cannot be excluded that “p. tuberoidea” and “p. aurata” are formed by very close (or even the same) taxa of fungi ............................... “polygonirhiza tuberoidea” 13. mycorrhiza with abundant emanating hyphae ................................................... 14 13*. mycorrhiza with smooth surface, no emanating hyphae observed ................. 15 14. small, yellowish mycorrhiza with very fine emanating hyphae. mantle surface covered with sand particles. mantle thin, plectenchymatous .................................................................................................... “polygonirhiza arenaria” 14*. mycorrhiza whitish-grey (older parts often pinkish-brown), big – reaching length of 3 mm. mantle plectenchymatous, formed by a dense hyphal net with matrix material. hyphae becoming thicker towards the inner layers of the mantle, without clamps. emanating hyphae abundant, often forming cottony concentrations, hyalin, thin-walled, with clamps, t-shaped branching, local inflations and frequent anastomoses. no colour reaction to koh, feso4 and melzer’s reagent ....................................................................................................... “polygonirhiza lanata” 15. mycorrhiza with whitish-creme colour. mantle thin (cortical cells visible) with smooth and shiny surface. outer mantle covered by a loose net formed by branched hyphae without clamps. mantle beneath net pseudoparenchymatous, epidermoid (fig. 2e). inner layer plectenchymatous ...................... “polygonirhiza epidermoidea” 15*. mycorrhiza whitish-grey (fig. 1a). mantle surface smooth and shiny, in older mycorrhizae cortical root cells visible. mantle thick, outer layer pseudoparenchymatous composed of roundish hyphal cells (fig. 2e), in inner layer hyphal segments of smaller diameter, less regular, and elongated. small mycorrhiza with a considerably big diameter and rounded top. mycelium not stained by koh, feso4 and sulphovanilin ................... “polygonirhiza lacteocinerea” quantitative aspects of ectomycorrhizal colonization. ectomycorrhiza was present in all analyzed samples. number of mycorrhizal tips differed strongly between sites (tab. 3). the richest specimens were characterized by m value near 0.2 (sites 1, 2, 5), whereas the quotient did not exceed 0.01 for plants from the site 3. 216 m. ronikier and p. mleczko average density of mycorrhizal tips for all analysed plants was almost equal in spring and autumn: 0.084 and 0.082 respectively. the ratio: alive vs dead mycorrhizae, was estimated for all sites. in all spring samples, number of living mycorrhizae was considerably lower than dead (ml/d < 1). this quotient was the lowest on the site 3 (ml/d < 0.16). the share of morphotypes in the total number of mycorrhizal tips was very differentiated. cenococcum geophilum and “polygonirhiza lacteocinerea” were clearly dominant, constituting appr. 23 % of all mycorrhizal tips each. “polygonirhiza vulpina”, “p. aspera”, “p. arenaria” and “p. fusca” represented 8–9 % of total number of ectomycorrhizae each. other morphotypes were less numerous, and sometimes limited only to few tips (“p. granulosa”, “p. aurata”, “p. terrea”, “p. maculata”). domination of cenococcum geophilum, “polygonirhiza lacteocinerea” and “p. vulpina” was correlated with their presence in all (in the case of two first) or most (3 – in case of the third) sites; to the contrary, “p. tuberoidea” and “p. arenaria” were limited to single sites only. some differences were observed in presence of morphotypes in samples collected in spring and autumn (sites 1, 2 and 3). five morphotypes on the site 1 (“p. aspera”, “p. rufocystidiata”, “p. radiata”, “p. salebrosa”, “p. tenua”) and two on the site 2 (“p. tuberoidea” and “p. aspera”) were present only in spring (in the case of “p. tuberoidea” it was connected with a clear domination of this morphotype in the samples). on the other hand, cenococcum geophilum was almost absent in spring, while in autumn it was very frequent in all sites. this was also true for “p. arenaria” on the site 1 (only 4 tips observed in spring and very abundant occurrence in autumn). these single observations are too scarce, however, to formulate any general conclusions. there were no clear differences between sites in the average number of mycorrhizae in relation to altitude. although the m value for the high mountain site 3 was low, the data for two other high-altitude stands (4 and 5) were higher and comparable with data for the lower located sites (1 and 2). endomycorrhiza in polygonum viviparum. prevailing part of roots did not manifest any traces of amf colonization, however, several roots taken from site 2 in spring contained intraradical structures resembling those formed by endomycorrhizal fungi (members of glomeromycota) – massive hyphae, coils and vesicles. no arbuscules, however, were observed. ta b l e 3 quantity of living and dead mycorrhizae (as m values with corresponding standard devia tions in brackets) and average numbers of morphotypes per plant on study sites site sampling period living mycorrhizae(m) dead mycorrhizae (m) average number of morphotypes on plant 1 spring 0.112 (0.068) 0.252 (0.102) 3.5 autumn 0.075 (0.034) 0.073 (0.023) 3.3 2 spring 0.100 (0.081) 0.130 (0.084) 3.0 autumn 0.086 (0.020) 0.045 (0.017) 4.0 3 spring 0.015 (0.008) 0.062 (0.032) 1.0 autumn 0.028 (0.025) 0.091 (0.068) 1.6 4 autumn 0.071 (0.014) 0.032 (0.017) 1.7 5 autumn 0.117 (0.063) 0.050 (0.027) 3.8 observations on the mycorrhizal status 217 discussion presence and diversity of ectomycorrhizae on polygonum viviparum in the tatra mts. seventeen ectomycorrhizal morphotypes were described in the samples from the tatra mts. comparable number was reported in the observations from italian alps (f o n t a n a 1977). the diversity of morphotypes on single specimens of p. viviparum reported from the alps also corresponds well with the situation in the tatra mts. f o n t a n a (1977) observed 2–3 morphotypes on average on a single plant, with maximum of 5 different mycorrhizae. the plants from denali national park (alaska) had at least 3 morphotypes each (tr e u et al. 1996). also in the material from rocky mts. “several morphotypes” were mentioned (m a s s i c o t t e et al. 1998). the universal presence of ectomycorrhizal colonization in p. viviparum on sites in whole altitudinal range of tatra mts. is in agreement with majority of observations. however, it does not correspond with some data, especially that of n e s p i a k (1953), who found specimens without any ectomycorrhizal colonization in two alpine sites in high tatra. however, comparison of data (from literature and present observations) for plants growing on different sites reveal rather small direct role of the position above sea level in the detected number of ectomycorrhizae, even though mycorrhizal colonization generally decreases with altitude (rewieved in k ö r n e r 1999). more probably, it could be suspected that the composition of the plant communities might have a strong influence on the ectomycorrhizal population of p. viviparum. a distinct, positive relationship was observed between the diversity of ectomycorrhizae of p. viviparum, and the presence of other ectomycorrhizal plants in its vicinity (cf. tab. 1). on plants growing near picea abies (l.) h. karst. (site 1), pinus mugo turra (site 2) or salix reticulata l. and dryas octopetala l. (site 5), the mycorrhizal colonization and/or the number of morphotypes was considerably higher than on specimens originating from the sites where p. viviparum was the only ectomycorrhizal plant (sites 3 and 4). similarly, in the paper by d o m i n i k et al. (1954) an abundant ectomycorrhizal colonization was reported for specimens from kominiarski wierch (1829 m) in the tatra mts., where p. viviparum grew together with pinus mugo, while lack of ectomycorrhiza in the study by n e s p i a k (1953) from the przełęcz białczańska pass (2080 m) in a patch of alpine grassland oreochloo distichae-juncetum trifidi could have resulted from lack of ectomycorrhizal plants. the presence of perennial, obligatorily mycorrhizal dwarf shrubs could play an important role in the creation and maintenance of the bank of ectomycorrhizal fungal inoculum (v ä r e et al. 1992). importance of this factor increases with the environmental stress at high altitude locations, diminishing the capability of fungi to grow and form fruitbodies. nevertheless, a quite high level of mycorrhizal colonization on the site 4 (although with only 3 morphotypes, two of them common for all the investigated sites), also devoid of ectomycorrhizal shrubs, suggests a notable autonomy of p. viviparum in the formation and maintenance of ectomycorrhiza. the presence of alive ectomycorrhizae in samples collected in spring and in autumn, together with a similar average number of mycorrhizal tips, suggest that the colonization is generally stable throughout the year. the absence of several morphotypes in spring could be the result of weaker ability to recolonize the roots after winter dormancy, however other reasons (eg. patchy distribution of mycelium) cannot be excluded. 218 m. ronikier and p. mleczko morphotypes recorded on polygonum viviparum. a comparison of ectomycorrhizae found in the tatra mts. with those from other sites is difficult, as very few descriptions are available. d o m i n i k et al. (1954) described an „ectotrophic mycorrhiza of the a type“; this type comprises a simple mycorrhiza without important ramifications nor growth modifications, with a primitive (plectenchymatous), loose mantle and hartig net of different depth (d o m i n i k 1961). the paper includes some general remarks on the mycorrhiza of p. viviparum, but does not allow comparison of morphotypes. one of dominant morphotypes in tatra mts. was formed by an ascomycete cenococcum geophilum. as a result of its commonness and a very characteristic appearance, the presence of this mycorrhiza is reported in most investigations from the whole distribution area of p. viviparum (e.g. f o n t a n a 1977; r e a d and h a s e l w a n d t e r 1981; tr e u et al. 1996; m a s s i c o t t e et al. 1998). it is not surprising, as the mycorrhiza of this fungus was described from many plant hosts (m a i a et al. 1996), and it was found on several other arctic-alpine species, as dryas octopetala, d. integrifolia, salix spp., kobresia belliardi (f o n t a n a 1963; tr a p p e 1964; r e a d , h a s e l w a n d t e r 1981; m a s s i c o t t e 1998). in lowlands cenococcum often dominates in dry environments (tr a p p e 1964). it is not the case in the mountains, characterized by relatively high falls and long snow depositions. as suggested by r e a d & h a s e l w a n d t e r (1981), the commonness of this fungus in such areas could be connected with an efficient spread strategy, that is the production of very resistant and abundant sclerotia, rather than any special symbiotic features. the high resistance of cenococcum to frost, experimentally demonstrated by c o r b e r y and l e ta c o n (1997) can also be important factor. the species from the genera amanita, inocybe and russula were also reported to form mycorrhiza with p. viviparum (rewieved in g a r d e s and d a h l b e r g 1996); an ectomycorrhiza of alnicola cholea and p. viviparum was also recently described (m o r e a u et al. 2006). a morphotype called “polygonirhiza lacteocinerea” strongly resembles an alpine mycorrhiza formed by russula nana ( russula emetica fr. var. alpestris boud.), described by f o n t a n a (1977). the presence of this mycorrhiza in tatra mts. is probable since this fungus is very common in the alpine belt there (n e s p i a k 1960; m . r o n i k i e r , pers. obs.). in the case of some morphotypes, many characteristics link them to the mycorrhizae described on trees. “polygonirhiza epidermoidea” share mantle features with e.g. the mycorrhizae of some russula spp. (epidermoid mantle structure with inner plectenchymatous layer, scarce extramatrical hyphae). a similar mycorrhiza is formed by russula firmula on pinus mugho (tr e u 1990). the morphotype “polygonirhiza aurata” seems to be close to the ectomycorrhiza of tomentella galzini described on quercus (j a k u c s et al. 1997 as „quercirhiza fibulocystidiata”; k õ l j a l g et al. 2001). both have an olive-green colour, pseudoparenchymatous, angular mantle structure and very characteristic cystidia with single clamps and thick walls below them. the only difference between these two morphotypes seems to be the lack of ramified emanating hyphae in the mycorrhiza of p. viviparum. also the features of the group of „black” mycorrhizae found on p. viviparum lead to suppose their possible relationships with several tree mycorrhizae, e.g. “piceirhiza nigra” (g r o n b a c h 1988), identified as formed by a member of thelephoraceae (a g e r e r et al. 1995). these morphotypes, having a pseudoparenchymatous mantle structure (“polygonirhiza aspera”, “p. rufocystidiata”, “p. salebrosa”, “p. fusca”), could be formed by this group of fungi. representatives of thelephoraceae forming dark mycorrhizae are mostly species observations on the mycorrhizal status 219 producing resupinate fruit-bodies on wood; interestingly, such morphotypes clearly dominated in sites in the vicinity of picea abies or pinus mugo (cf. tab. 1, 2), so they could be formed not by alpine fungi but species related with trees – tomentella spp. it is not possible, however, to refer such an assumption to available mycological data from the tatra mts. as this group of fungi has not been studied in this area so far. a very characteristic star-like hyphae arrangment and the presence of dark, incrusted emanating hyphae in “polygonirhiza aspera” resembles strongly “fagirhiza setifera” (b r a n d 1991), however the fagus mycorrhiza has abundant cystidia, lacking in the p. viviparum mycorrhiza. the general morphological aspects of ectomycorrhiza formed by p. viviparum in the tatra mts. fit well descriptions by b l a s c h k e (1991a), tr e u et al. (1996) and m a s s i c o t t e et al. (1998). it may be concluded that this species forms exclusively simple, cylindrical or club-shaped mycorrhizae. the anatomical features of these mycorrhizae, particularly the characteristic growth modification of epidermal cells (m a s s i c o t t e et al. 1998), are similar to mycorrhizae of other angiosperms, including trees, e.g. beech (a g e r e r 1991; s m i t h , r e a d 1997). presence of arbuscular mycorrhiza in polygonum viviparum. traces of probable arbuscular mycorrhizal colonization were sporadically observed. the roots contained some characteristic structures typically associated with the arbuscular mycorrhiza, as hyphae, coils and vesicles, but they were not accompanied by arbuscules. most observations of p. viviparum reported lack of endomycorrhizal colonization, although it was incidentally found in arctic sites (s t a h l 1900) as well as in the mountains (n e s p i a k 1953; b l a s c h k e 1991a, 1991b). n e s p i a k (1953) noticed exclusively endomycorrhizal colonization without ectomycorrhizae in the same root system, while b l a s c h k e (1991a, 1991b) found both kinds of symbiosis occurring together. none of these authors, however, mentioned the formation of arbuscules in the roots. the infection of non-host roots by am fungi, including formation of vesicles, was reported in some cases; the main signal which controls the development of functional symbiosis probably acts by trigerring fungal genes responsible for change of hyphal growth and physiology during arbuscule formation (review in g i o v a n n e t t i and s b r a n a 1998). considering the presence of some am structures in p. viviparum roots, the capacity of this plant to form this kind of symbiosis seems to be probable even if not important ecologically. lack of arbuscules could possibly be also due to their short-lived appearance during the vegetation period, as it was reported in the study of the high-mountain ranunculus adoneus colonization by glomus tenuis (m u l l e n , s c h m i d t 1993). regular phenological study or controlled cultures would be necessary to verify potential factors responsible for establishment of arbuscular mycorrhiza in p. viviparum. the present study is the first contribution focused on the mycorrhiza of polygonum viviparum in the tatra mts. and the carpathians. the results showing that ectomycorrhizal colonization is a regular situation in this species, but affected by several factors, should be the starting point for future studies employing rigorous morphological/anatomical descriptions of morphotypes, regular survey of carpophores on permanent plots and employing dna comparisons of mycorrhizae and carpophores. including comparative analysis of diversity of mycorrhizae in neighbouring ectomycorrhizal plants in plant communities with p. viviparum would al220 m. ronikier and p. mleczko low a direct estimation of share/independence of the ectomycorrhizal diversity of p. viviparum. acknowledgments: this paper is based on an msc project carried out by m. ronikier under supervision of prof. katarzyna turnau (jagiellonian university, kraków). the authors thank prof. k turnau for help ful discussions and comments, and dr. anna ronikier for help in gathering plant samples and comments on the manuscript. references a g e r e r r. 1986. studies on ectomycorrhizae ii. introducing remarks on characterization and identification. mycotaxon 26: 473 492. a g e r e r r. 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(in:) w. s z a f e r , k. z a r z y c k i (eds). szata roślinna polski. 1. pwn, warszawa: 129 206. p a w ł o w s k i b. 1956. flora tatr. rośliny naczyniowe. 1. pwn, warszawa: 204 205. p e y r o n e l b. 1930. simbiosi micorrizica tra piante alpine e basidiomiceti. nuovo giorn. bot. ital. 37: 655 663. p e y r o n e l b. 1937. osservazioni e considerazioni sul fenomeno della micorriza al piccolo san bernardo. nuovo giorn. bot. ital. 44: 587 594. p h i l l i p s j., h a y m a n d. s. 1970. improved procedures for clearing roots and staining parasitic and vesicu lar arbuscular mycorrhizal fungi for rapid assessment of infection. trans. br. mycol. soc. 55: 158 161. r e a d d. j., h a s e l w a n d t e r k. 1981. observations on the mycorrhizal status of some alpine plant communi ties. new phytol. 88: 341 352. s m i t h s. e., r e a d d. j. 1997. mycorrhizal symbiosis. academic press. s t a h l e. 1900. der sinn der mycorrhizenbildung. eine vergleichend biologische studie. jahrb. wissenschaft. bot. 34: 539 668. t r a p p e j. m. 1964. mycorrhizal hosts and distribution of cenococcum graniforme. lloydia 27: 100 106. t r e u r. 1990. charakterisierung und identifizierung von ectomycorrhizen aus dem nationalpark berchtesga den. bibl. mycol. 134: 80 84. t r e u r., l a u r s e n g. a., s t e p h e n s o n s. l., l a n d o l t j. c., d e n s m o r e r. 1996. mycorrhizae from denali national park and preserve, alaska. mycorrhiza 6: 21 29. v ä r e h., ve s t b e r g m., e u r o l a s. 1992. mycorrhiza and root associated fungi in spitsbergen. mycor rhiza 1: 93 104. obserwacje statusu mikoryzowego polygonum viviparum w polskich tatrach (karpaty zachodnie) s t r e s z c z e n i e polygonum viviparum jest jednym z nielicznych gatunków roślin zielnych, które tworzą ektomikoryzę. w tatrach rdest żyworodny należy do gatunków dominujących w piętrze alpej skim, występuje również niżej sięgając do podnóży masywu. celem badań była wstępna ana liza różnorodności ektomikoryz tworzonych przez ten gatunek w tatrach oraz ogólna analiza jej zależności od warunków ekologicznych takich jak wysokość nad poziom morza oraz skład zbiorowisk roślinnych. korzenie p. viviparum były również dodatkowo badane pod kątem obecności kolonizacji endomikoryzowej. 222 m. ronikier and p. mleczko próby korzeni zebrano z 5 stanowisk na podłożu granitowym i wapiennym, rozmieszczo nych w przedziale wysokości 900 2150 m n.p.m. ektomikoryzy były obecne na wszystkich badanych okazach polygonum viviparum; na pojedynczych roślinach obserwowano 2 5 mor fotypów. w sumie zaobserwowano i krótko scharakteryzowano 17 morfotypów ektomikoryz. najbardziej rozpowszechnione we wszystkich próbach były mikoryza cenococcum geophilum oraz niezidentyfikowany, jasno zabarwiony morfotyp przypominający mikoryzy russula sp. nie stwierdzono znaczących różnic w poziomie kolonizacji ektomikoryzowej pomiędzy stano wiskami różniącymi się położeniem nad poziomem morza. zaobserwowane różnice w liczeb ności i różnorodności mikoryz p. viviparum na poszczególnych stanowiskach wiązać można najprawdopodobniej ze składem gatunkowym zbiorowisk roślinnych obecnością krzewinek ektomikoryzowych spełniających zasadniczą rolę w utrzymywaniu populacji grzybów ektomi koryzowych. należy jednak podkreślić, że ektomikoryzy obserwowano również na stanowi skach, gdzie p. viviparum było jedynym potencjalnym symbiontem ektomikoryzowym. regularnie obserwowano kolonizację korzeni polygonum przez grzyby endofityczne. w kilku korzeniach odnotowano obecność struktur charakterystycznych dla mikoryzy arbu skularnej (pęcherzyki, peletony), jednak nie towarzyszyły im wykształcone arbuskule. 2014-01-01t11:44:17+0100 polish botanical society 2014-09-02t20:10:26+0200 polish botanical society agaricus chionodermus pilát, a species new to bulgaria 199this is an open access article distributed under the terms of the creative commons attribution 3.0 license (creativecommons.org/licenses/by/3.0/), which permits redistribution, commercial and non-commercial, provided that the article is properly cited. © the author(s) 2014 published by polish botanical society acta mycologica introduction the genus agaricus comprises a diverse, cosmopolitan saprotrophic agarics occurring in a variety of ecosystems. most of them, however, are confined to grasslands [1–4]. there are ca. 200 described agaricus species worldwide and approximately 60 of them occurring in europe [4–8]. there have been reported 54 species of the genus from bulgaria so far [9–17]. five of the agarics are included in the “red list of fungi in bulgaria” [18]. in the course of studies of the diversity of genus agaricus in bulgaria, a. chionodermus was found by the author for the first time. it is presumably a rare species for many countries of europe and yet not well known. therefore in this paper a detailed description and illustrations are presented including features that distinguish a. chionodermus from similar species. original research paper acta mycol 49(2):199–205 doi: 10.5586/am.2014.019 received: 2014-02-21 accepted: 2014-10-16 published electronically: 2014-12-11 agaricus chionodermus pilát, a species new to bulgaria maria lacheva* department of botany and agrometeorology, agricultural university-plovdiv 12, mendeleev, 4000 plovdiv, bulgaria abstract the article presents the first record of agaricus chionoderma pilát in bulgaria. basidiomata of the species were found on june 2004, september 2010, and september 2011 in the thracian lowland floristic region. the article brings closer taxonomic profile, ecological requirements and distribution pattern of the species. differences from similar species are discussed. it also describes macroscopic and microscopic characteristics of the discovered specimens and presents the specification of habitat the fungus concerned. the species are described and illustrated by photographs of macroscopic and microscopic features on the basis of bulgarian specimens. keywords: agaricomycetes; agaricus sect. arvenses; bulgarian mycobiota; fungal diversity; taxonomy * email: agaricus@abv.bg handling editor: maria rudawska http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.2014.019 mailto:agaricus%40abv.bg?subject=am.2014.019 200© the author(s) 2014 published by polish botanical society acta mycol 49(2):199–205 lacheva / agaricus chionodermus pilát a species new to bulgaria this work adds a new species to the bulgarian agarics, namely a. chionodermus pilát, one of the about seventy “european” species which, according to cappelli [4], belong to agaricus sect. arvenses. material and methods air-dried studied specimens of the fungus are kept in the mycological collection of the agricultural university-plovdiv (soa). basidiomata of species were photographed with sony cyber-shot. description of morphological characters of basidiomata is based on fresh and dried specimens. micromorphological features were observed and measured in fresh and dry fragments of tissues dehydrated in water, under an amplival ml light microscope, with magnification ×1000. microphotographs were taken on amplival ml. drawings were made with the aid of a drawing tube under an oil-immersion objective. spores were examined in melzer’s reagent and were taken from the spore print for measurements. calculated values are based on 50 measurements. the obtained data for length and width of microstructures were examined by standard statistic methods [19,20] and are presented in the text in the following way: (min–) mean ±1σ (–max), n = 50, (l/wmin–max); of the basidia and cheilocystidia: min–max. schäeffer reaction was tested by aniline and 65% hno3 acid on dried samples [21]. the abbreviations of the author names of fungal species are given according to kirk and ansell [22]. nomenclature of species follows kirk et al. [8]. the taxonomic and nomenclature decisions in the article have been made in conformity with the most appropriate taxonomic monographs of the agaricus genera [4,23,24]. the chorological map of the occurrence of the a. chionodermus in the country, have been depicted using the program software dsoa [25]. results and disscusion agaricus chionodermus pilát has never been previously found in bulgaria [12,26]. the occurrence of first a. chionodermus basidiomata was discovered on 7th june 2004 in thracian lowland, plovdiv distr., above stryama village (fig. 1). the area where the fungus occurred is situated along the watercourse of stryama river and is surrounded by farmland, as well as meadows and pastures. the dominant species of the tree layer at the plant community where basidiomata of a. chionodermus has been found were: acer campestre, fraxinus angustifolia, populus canescens and tilia cordata. the shrub layer includes mainly: populus tremula, salix caprea, corylus avellana and sambucus nigra. the herb layer was dominated by cereal grasses, e.g. carex caryophyllea, poa pratensis, etc. in the course of the field trial only three specimens of a. chionodermus were found. the basidiomata of the agarics were revealed in grassy vegetation, close to p. canescens (ait.) sm. the next finding of basidiomata a. chionodermus was done in the same place in september 2010 and september 2011. the location of the species has been carefully marked and will be monitored in the future. 201© the author(s) 2014 published by polish botanical society acta mycol 49(2):199–205 lacheva / agaricus chionodermus pilát a species new to bulgaria species description agaricus chionodermus pilát (fig. 2, fig. 3), acta mus. nat. prag., vii b, 1: 134, 1951. icones: pilat (1951: fig. 60; tab. 3, tab. 14–16), pilat (1953: fig. 22), cappelli (1984: тab. 48). macroscopic and microscopic features. pileus up to 7.5–12(–15) cm in diameter, thick-fleshed, initially hemispherical, subsequently convex to plano-convex, seldom flat or slightly depressed, silky finely fibrillose, sometimes cracked dry in the center, snow-white, white, sometimes greyish white in the center, slightly yellowish towards the margin. margin undulate, initially involute, thin, with whitish silky fibrillose scales, 5–7 mm thick, finally with remnants of the partial veil. pileipellis consisting of whitish thick-walled cylindrical hyphae, with clamps, 5–10 μm in diameter. gills free, thin, crowded, narrowing toward the margin of pileus, long, with an even edge, whitish-pink, subsequently light pink, gray-pink to dark-brown, with light, sterile edge. hymenophoral trama in young basidiomata initially regular, latter irregular, consisting of cylindrical, thin-walled hyphae, 5–12 μm in diam. stipe up to 8–15 × 1.5–3 cm, central, cylindrical, with or without slightly bulbous base, initially white, subsequently ± white, smooth to silky-fibrillose, yellowish at touch. flesh whitish, on cutting becoming lemon-yellow in the base of the stipe. ring wide apical, fleecy, free standing, whitish, with touch upper layer yellowish tinge and thick, delicate, sericeous, bottom layer. context in pileus and stipe fragile, whitish. smell non-distinctive, very faint, non of almonds. taste non-distinctive. spores 7.5–(4.2 ±0.02)–10 × 4.5–(4.1 ±0.02)–6 μm, (n = 50), ellipsoid to ovoid, brown, not ornamented, with fluorescent spots, with an apical germ pore. spore print dark brown. basidia 20–28 × 8.5–10.5 μm, clavate, hyaline, 4-sterigmate. sterigmata 2–3 μm long. cheilocystidia 17–30 × 6.5–10.5 μm, crowded, clavate to cylindrical, lageniform, thinwalled, hyaline. macrochemical reactions: cross reaction with schaeffer’s reagent: positive. fig. 1 distribution of agaricus chionodermus in bulgaria. 202© the author(s) 2014 published by polish botanical society acta mycol 49(2):199–205 lacheva / agaricus chionodermus pilát a species new to bulgaria specimens examined. bulgaria, thracian lowland, plovdiv distr., above stryama village, right slope, alt. 240 m a.s.l., on mown meadow close to populus canescens (ait.) sm. and salix caprea l., along the watercourse of stryama river, near by the bridge to the river, on basic soil, 7 june 2004, coll. & det. m. lacheva (soa 60 00292); ibid., on sandy soil, 30 september 2010 (soa 60 00293), 14 september 2011, coll. & det. m. lacheva (soa 60 00367). taxonomical remarks. nearly identical basidiomata produces a. arvensis schäeffer known to be common in europe, which can be separated with certainty from a. chionodermus by the cap being generally snow-white, wide sericeous ring and the slightly larger spores [1,4,23], but mainly on the base of microscopic characters. as emphasized by cappelli [4], a. chionodermus in contrast to, a. arvensis has round and conspicuous cheilocystidia. moreover, there is an ecological difference between the two species, i.e. a. chionodermus grows on periphery of deciduous and coniferous forest, or forest meadows, on calcareous and sandy soils, while a. arvensis occurs on humus rich soil, mainly among grasses. agaricus nivescens (f.h. møller) f.h. møller also resembles a. chionodermus, from which it mainly differs by habitat (terrestrial fungus occurs mainly in grasslands on calcareous soil), almonds and fungoid smell, indeterminate taste, and smaller size of spores and basidiomata [4,27]. metric and macroscopic data of bulgarian specimens of a. chionodermus agree with the descriptions of [4] and [23]. according to this investigation, a. chionodermus should be considered as a species developing in grassy vegetation on forest meadows at lower elevation, close to poplar, possibly also under other broadleaved tree species. the observations are in accordance with the literature data that a. chionodermus should be considered as species developing mostly on basic to neutral or sandy soil [4]. habitat, ecology, phenology, edibility status. basidiomata of a. chionodermus almost exclusively appears in summer to early fall season (may–october), solitary, in periphery of deciduous forest or forest meadows and grassy vegetation, 200–700 m alt. humus saprotroph, mostly on basic to neutral or sandy soil. there are some information that this species is edible without gastrointestinal problems [4,28]. there fig. 2 basidiomata of agaricus chionodermus at different stages of development in situ from bulgaria (photos: m. lacheva). 203© the author(s) 2014 published by polish botanical society acta mycol 49(2):199–205 lacheva / agaricus chionodermus pilát a species new to bulgaria is so far no indication in bulgaria that its edibility is known and presently the fungus is unlikely to be collected for food, beside if not be confused with other similar species. general distribution. agaricus chionodermus has a relatively wide distribution. the species has been recorded in many european countries, but in some regions it is regarded as rare. in europe, a. chionodermus is reported from czech republic [23,29], france [27,30], italy [4,31], spain [32,33] and ukraine [1]. in asia the species has been recorded in turkey [34–36]. conclusion agaricus chionodermus should be especially sought after in plains and lowlands regions of the country, but its presence in other parts of bulgaria cannot be excluded. basidiomata of this fungus might have been misidentified because of its resemblance to other similar agaricus species from section arvenses, especially if a spore print was not available. this new finding contributes to the diversity of bulgarian mycobiota, by adding new agaricus records and might be useful for creating a database of the bulgarian sabulicolous agaricus species. fig. 3 microscopic features of agaricus chionodermus from bulgaria: basidiospores (a), basidia (b), cheilocystidia (c), generative hyphae of the pileipellis with terminal elements (d). photos and drawings: m. lacheva. scale bars: 2.5 µm (a), 5 μm (b–d). acknowledgments the author expresses gratitude to assoc. prof. melania gyosheva (department of plant and fungal diversity and resources, institute of biodiversity and ecosystem research, bulgarian academy of sciences, sofia, bulgaria) for critically reading the manuscript. the author thanks two anonymous reviewers for useful comments. references 1. wasser sp. flora fungorum rss ucrainicae. kiev: naukova dumka; 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(agaricales, basidiomycotina) of the iberian peninsula and balearic islands. cuad trab flora micolόgica ibérica. 2005;21:1–101. 25. stoyanov k. documentation system in herbarium of agricultural university of plovdiv, bulgaria. j balk ecol. 2003;6(1):28–34. 26. denchev cm, assyov b. checklist of the larger basidiomycetes in bulgaria. mycotaxon. 2010;111(1):279– 282. http://dx.doi.org/10.5248/111.279 27. gerault a. florule evolutive des basidiomycotina du finistere. homobasidiomycetes. agaricales. version 2.1; 2005. 28. gminder a. ständerpilze: blätterpilze iii. in: krieglsteiner gj, gminder a, editors. die großpilze badenwürttembergs. stuttgart: eugen ulmer kg; 2010. p. 671. 29. pilát a. hymenomycetes novi vel minus cogniti cechoslovakiae: 2. acta mus nat prag. 1953;9b(2):1–140. 30. heinemann p. essai d’une clé de determination des genres agaricus et micropsalliota. sydowia. 1977;30(1–6):6–34. 31. cappelli a. il genere agaricus l. ex fr. s. karsten. sezione “rubescentes” dei bischi. boll grup micol g bresadola trento. 1983;26(1–2):4–38. http://dx.doi.org/10.1128/aem.66.2.728-734.2000 http://dx.doi.org/10.1128/aem.66.2.728-734.2000 http://dx.doi.org/10.5248/111.279 205© the author(s) 2014 published by polish botanical society acta mycol 49(2):199–205 lacheva / agaricus chionodermus pilát a species new to bulgaria 32. fernández jmr. guía micológica: tomo 3: género agáricus en españa. bilbao: jmr fernández; 1997. 33. moreno a, remondo j. flora micolόgica de la rioja. zubía. 1999;17:11–43. 34. solak mh, kalmıs e, isıloğlu m. new records for the fungi flora of turkey. bio-sci res bull. 2001;17:99–103. 35. solak mh, isıloğlu m, kalmıs e, allı h. macrofungi of turkey. checklist. bornova: universiteliler ofset; 2007. (vol 1). 36. sesli e, denchev cm. checklists of the myxomycetes, larger ascomycetes, and larger basidiomycetes in turkey. mycotaxon. 2008;106:65–68. abstract introduction material and methods results and disscusion species description conclusion references 2014-12-31t17:26:38+0000 polish botanical society 2014-11-20t23:09:20+0000 polish botanical society 2014-11-20t23:10:20+0000 polish botanical society 2014-11-20t23:10:43+0000 polish botanical society 2014-09-03t19:46:31+0200 polish botanical society 2014-09-03t19:47:06+0200 polish botanical society 2014-09-03t19:46:44+0200 polish botanical society 2014-09-06t20:22:27+0200 polish botanical society 2014-09-03t19:47:23+0200 polish botanical society 2014-09-02t20:10:53+0200 polish botanical society leucopaxillus lepistoides, a new steppe fungus in poland janusz łuszczyński department of botany, institute of biology, świętokrzyska academy świętokrzyska 15, pl 25 406 kielce, jluszcz@pu.kielce.pl ł u s z c z y ń s k i j .: leucopaxillus lepistoides, a new steppe fungus in poland. acta mycol. 41 (2): 279 284, 2006. the paper presents information on leucopaxillus lepistoides (maire) singer, a new species for poland. this fungus was found in two localities: the neighbourhood of busko zdrój and chęciny (little polish upland, s poland). both localities were in the xerothermic grasslands belonging to the cirsio brachypodion order, festuco brometea class. key words: leucopaxillus lepistoides, xerothermic grasslands, steppe fungi, thermophilous fungi introduction the climatic-habitat conditions occurring in poland, are generally adverse for developing of thermophilous, steppe sensu stricto, and southern-european species. only some species with above mentioned characters find appropriate conditions to grow, and only on relatively small areas of southern slopes limestone or gypsum hills where existing microclimate simulates a warm and dry mediterranean or continental climate. occurrence of steppe fungi among polish macrofungi biota is relatively rare. some informations about fungi of this ecological group derive mainly from wyżyna małopolska (little polish upland), and pomerania, occupied by xerothermic grasslands (b u j a k i e w i c z 1979; ł u s z c z y ń s k i , ł u s z c z y ń s k a 1991(1992), 2006 (in press); s t a s i ń s k a 2003; s t a s i ń s k a , p r a j s 2002; š m a r d a 1957; wo j e w o d a 1975). a new site of leucopaxillus lepistoides, a typically steppe fungus on the northen limit of the steppe plants range and xerothermic grasslands is worth mentioning. acta mycologica vol. 41 (2): 279-284 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 280 j. łuszczyński species description leucopaxillus lepistoides (maire) singer z. pilzk., 17: 14. 1939 – tricholomataceae, agaricales, agaricomycetidae, basidiomycetes, basidiomycota, fungi (k i r k et al. 2001). syn.: tricholoma lepistoides r. maire this species consists of two varieties: leucopaxillus lepistoides var. lepistoides, and leucopaxillus lepistoides var. pannonicus bohus. l. lepistoides var. lepistoides has smooth spores, whereas l. lepistoides var. pannonicus is characterized by rough spores. morphology. fruit-body consists of pileus and stipe. cap 15-25 cm diam, when young hemisphaere convex with involuted margin, white or creme, when old flattened, in the middle grayish, gray-brownish, cracked on areolas. white, indented lamellae adnated to stipe, or slightly decurrent. stipe cylindric, in the lower part slightly widen, relatively short, massive, 5-9 x 4-6 cm, the same colour as cap, mature with blue-greenish shade, when rubbed and/or damaged changing to intensive bluegreenish colour. odour when fresh unspecific, sometimes floury, but in dry specimens strongly magghi. spores in powder creme, under the microscope colourless, broadly elipsoid, smooth, and weakly amyloid 8,1-10,4 x 5-6,5 μm (fig. 1). the collected material was deposited in the herbarium of the department of botany, institute of biology, świętokrzyska academy, kielce (ktc 3860, 3861). localities in poland. until now leucopaxillus lepistoides was found in poland only on two (fig. 2). the first one: the nida basin, wola zagojska village, 6 km sw of busko zdrój, and ca. 50 km s of kielce, atpol square fe 24, in the xerothermic community of adonido-brachypodietum. the patch where the fruit-bodies were found (09.07.1991, coll. j. łuszczyński), was situated on the 25o inclined slope with the sw exposition, on the gypsum rendzinas. the floristic composition of this phytocoenosis was as follows: cover of herb layer 100%, brachypodium pinnatum 2.3, campanula sibirica +, plantago media +.2, seseli annuum +, astragalus danicus +.2, teucryum chamaedrys 5.5, festuca valesiaca +.2, euphorbia cyparissias 2.1, achillea pannonica 1.1, asperula cynanchica +.2, carex humilis 1.2, festuca rupicola 1.2, thymus kosteleckyanus 1.2, agropyron intermedium +, galium verum +, medicago falcata +.2, coronilla varia +, poa angustifolia +, plantago lanceolata +, inula ensifolia +, agrimonia eupatoria +. fig. 1. leucopaxillus lepistoides: spores; bar = 10 μm. leucopaxillus lepistoides, a new steppe fungus 281 the second site, the polichno village, 8 km wwn of chęciny, and ca. 17 km sww of kielce, atpol square ee 82, in the xerothermic grassland of thalictrosalvietum (27.06.2004, coll. j. jaworska). fruit-bodies were found, on the 30o inclined slope with the s exposition, on the rendzinas orginating from jurassic limestone. the floristic composition of this phytocoenosis is as follows: cover of herb layer 100%, moss layer 15%: brachypodium pinnatum 4.4, plantago media 1.1, prunella grandiflora 2.2, anthericum ramosum +, agropyron intermedium ssp. trichophorum +, achillea pannonica 1.1, campanula sibirica 1.1, potentilla arenaria 2.2, scabiosa ochroleuca 2.2, asperula cynanchica 2.2, seseli annuum 1.1, anthyllis vulneraria 1.1, euphorbia cyparissias 1.1, dianthus carthusianorum +, helianthemum nummularium ssp. obscurum 2.2, pimpinella saxifraga 1.1, salvia pratensis 1.1, abietinella abietina d 2.2, veronica spicata 1.1, carex caryophyllea +, galium verum 1.1, galium album 1.1, silenie otites +, hieracium pilosella 1.1, hypericum perforatum 1.1, festuca rupicola 1.1, knautia arvensis +, medicago lupulina 1.1, trifolium pratense 1.1. distribution. moser (1973) defines leucopaxillus lepistoides as a typically steppe species. it is known mainly from southern europe: bulgaria, czech republic, germany, greece, france, hungary, italy, romania, slovakia, spain (b o h u s 1966; c a m b o n i , m i g l i o z z i 2001; g a l l i 1994; h i n k o v a , s t o i č e v 1983; l i z o ň 2001; m i s k y et al. 2003; www.manitari.gr/manitaria/list/l.htm), but also from africa (libya, mauretania), middle america (costa rica), and asia (china; http://info. kib.ac.cn/soft/2286.htm). fig. 2. distribution of leucopaxillus lepistoides in poland. 282 j. łuszczyński discussion leucopaxillus lepistoides although reported from several european countries, is considered to be a very rare fungus. in poland the bigest threat for this species may arise from the secondary succesion of xerothermic grasslands and changing relations among many ecological factors, like light, temperature, humidity, and changing of substratum. it should be included in the polish red list of threatened fungi, in category en. in a few countries this species is on the red list data book or/and on list of fungi to be protected by law, for example in slovakia (l i z o ň 2001), and czech republic (f e l l n e r 2005). acknowledgments. the author would like to express great thanks to miss justyna jaworska for allowing to use her collection of leucopaxillus lepistoides. references b o h u s g . 1966. leucopaxillus arten in ungarn. (a leucopaxillus nemzetség magyarországi fajai). fragm. bot. mus. hist. nat. hung. 4: 33 40. b u j a k i e w i c z a . 1997. macromycetes occurring in the violo odoratae ulmetum campestris in the bielinek reserve on the odra river. acta mycol. 32 (2): 189 206. c a m b o n i m., m i g l i o z z i v. 2001. la micoflora del litorale romano. 6^o contributo. alcune specie comuni. hygrophorus roseodiscoideus, leucopaxillus lepistoides, rhodocybe gemina e rhodocybe ge mina var. subvermicularis. micologia italiana 30 (3): 60 72. f e l l n e r r . 2005. conservation of fungi in the czech republic: situation to the year 2005. european council for the conservation of fungi. newsletter 14: 7 10. g a l l i r . 1994. una specie rara mediterranea: leucopaxillus lepistoides. micologia e vegetazione me diterranea 9 (1): 20 24. h i n k o v a c . , s t o i č e v g . 1983. leucopaxillus lepistoides (maire) sing. in bulgaria. third nac. conf. of botany, с., ban: 39 42. l i z o ň p. 2001. red list of slovak fungi. (http://www.wsl.ch/eccf/slovakia.pdf). ł u s z c z y ń s k i j . , ł u s z c z y ń s k a b . 2006. bioróżnorodność grzybów basidiomycetes w fitocenozach kserotermicznych okręgu chęcińskiego i problemy ich ochrony (in press). ł u s z c z y ń s k i j . , ł u s z c z y ń s k a b . 1991 (1992). nowe stanowiska gasteromycetes w okolicy buska zdroju. acta mycol. 27 (2): 221 223. m i s k y m . , k o v á c s j . , a l b e r t l . , b r a t e k z . 2003. knowledge of fungi in surroundings of cristuru secuiesc i. macrofungi. moeszia. (http://www.ngo.ro/lkg/moeszia.html) m o s e r m . 1983. die röhlinge und blätterpilze (polyporales, boletales, agaricales, russulales). kleine kryptogamenflora ii b/2, basidiomyceten 2. veb g. fischer verl., jena. 532 pp. š m a r d a j . 1957. přispěvek k poznání gasteromycetů v polsce. acta soc. bot. pol. 26: 319 324. s t a s i ń s k a m . , p r a j s b . 2002. new record of montagnea arenaria (fungi, agaricales) and its distri bution in poland. polish bot. j. 47 (2): 211 213. s t a s i ń s k a m . 2003. różnorodność grzybów (macromycetes) w warunkach naturalnej sukcesji mu raw stepowych: 31 34. (in:) s. m. r o g a l s k a , j. d o m a g a ł a (ed.). człowiek i środowisko przy rodnicze pomorza zachodniego. i środowisko biotyczne. uniw. szczecin. wyd. oficyna in plus. szczecin. w o j e w o d a w. 1975. macromycetes ojcowskiego parku narodowego. ii. charakterystyka socjologicz no ekologiczno geograficzna. acta mycol. 11 (2): 163 212. leucopaxillus lepistoides, a new steppe fungus 283 leucopaxillus lepistoides, nowy grzyb stepowy w polsce s t r e s z c z e n i e ogólne uwarunkowania klimatyczno siedliskowe panujące w polsce mało sprzyjają roz wojowi stepowych i południowo europejskich gatunków. tylko nieliczne gatunki o takim charakterze ekologicznym i geograficznym mogą rozwijać się na szczególnie eksponowanych siedliskach, wapiennych i gipsowych wzgórz. na ciepłych i suchych siedliskach kserotermicz nych występują różne gatunki grzybów zdolne znieść specyficzne warunki takich siedlisk ale grzybów o charakterze stepowym sensu stricto w polsce mamy nie wiele. spotykane bywają tylko na pojedynczych stanowiskach na wyżynie lubelskiej, małopolskiej, pomorzu zachod nim i środkowym. w okolicach buska zdroju, we wsi wola zagojska (niecka nidziańska) i w okolicach chęcin, we wsi polichno (góry świętokrzyskie), w murawach kserotermicznych należących do zespołów adonido brachypodietum i thalictro salvietum, znaleziono dwa sta nowiska nowego dla polski grzyba leucopaxillus lepistoides. gatunek ten interesujący jest również z uwagi na jego wymagania ekologiczne, gdyż uważany jest za typowy element ste powy (m o s e r 1973). nietrwały, w naszej szerokości geograficznej, charakter muraw ksero termicznych i możliwość zarastania ich przez zarośla krzewiaste stanowią poważne zagrożenie dla tego grzyba. w związku z powyższym proponuje się włączyć go do czerwonej listy grzybów zagrożonych w polsce w kategorii wymierające (en). 2014-01-01t11:44:35+0100 polish botanical society 2014-11-20t23:12:46+0000 polish botanical society 2014-09-08t11:11:13+0200 polish botanical society 2014-09-05t16:22:26+0200 polish botanical society 2014-09-05t16:24:08+0200 polish botanical society © the author(s) 2014 published by polish botanical society the preservation status of the lichen biota in the designed special area of conservation natura 2000 „middle łyna river valley – smolajny” dariusz kubiak1, paweł czarnota2, anna zduńczyk3, maria dynowska1, grzegorz leśniański4, aleksandra cygańska5, sandra olszewska6, anna sadowska-deś7 and paweł wojdal8 1department of mycology, warmia and mazury university in olsztyn oczapowskiego 1a, pl-10-719 olsztyn, darkub@uwm.edu.pl 2department of agroecology, faculty of biology and agriculture university of rzeszów, ćwiklińskiej 2, pl-35-601 rzeszów, pawczarnota@poczta.onet.pl 3department of botany and ecotoxycology, institute of biology and environmental protection pomeranian university in słupsk, arciszewskiego 22b, pl-76-200 słupsk 4laboratory of lichenology e3lichlab, w.m. bartla 19c/61, pl-30-389 kraków 5inter-faculty studies in environmental protection, university of warsaw żwirki i wigury 93, pl-02-089 warsaw, grabowskaaz@wp.pl 6department of plant taxonomy and nature conservation, gdańsk university wita stwosza 59, pl-80-308 gdańsk, sandra_strahl@wp.pl 7biodiversity and climate research centre bik-f, senckenberganlage 25 d-60325 frankfurt am main, anna-d-sadowska@o2.pl 8laboratory of nature protection, faculty of biology, university of warsaw miecznikowa 1, pl-02-096 warsaw, bocian.freestyle@gmail.com kubiak d., czarnota p., zduńczyk a., dynowska m., leśniański g., cygańska a., olszewska s., sadowska-deś a., wojdal p.: the preservation status of the lichen biota in the designed special area of conservation natura 2000 „middle łyna river valley – smolajny”. acta mycol. 49 (1): 135–146, 2014. the paper presents the list of 159 taxa, including 151 lichens and 8 saprotrophic or parasitic (lichenicolous) fungi, recorded in the designed special area of conservation natura 2000 „middle łyna river valley – smolajny” (the forest division of wichrowo). the analysed area (2953 ha) covers mostly forest communities, with natural character, associated with the valley of the łyna river (hillside lime-oak-hornbeam forests, streamside alder-ash forest, riparian black alder forest). key words: lichens, lichenized fungi, species diversity, forest acta mycologica vol. 49 (1): 135–146 2014 doi: 10.5586/am.2014.010 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 136 d. kubiak et al. © the author(s) 2014 published by polish botanical society introduction nature conservation is one of the state policy aspects defined in the nature conservation act (dziennik ustaw 2004) and in the related executive regulations. one of the nature conservation objectives, within the meaning of the aforementioned act, is to preserve the biodiversity of the country. any measures aimed at active or passive protection of species diversity require comprehensive knowledge of habitat requirements and distribution of species from different taxonomic groups. furthermore, measures of this kind are a prerequisite for the assessment of potential threats to individual taxa. it is estimated that over 65% of biological resources in poland are present in forests (jaroszewicz 2007). therefore, in view of efficient management of the polish fauna, flora and fungi resources, the knowledge about nature resources of forest ecosystems appears to be extremely important. the state forests are obliged by the forest act (ustawa 1991) to initiate, coordinate and conduct periodic assessments of the broadly defined forest resources. in practice, this type of comprehensive assessment is usually not possible due to the lack of professionals adequately trained. the paper presents the results of lichenological research conducted within the framework of field sessions accompanying the 25th convention of polish lichenologists „lichens in the geographic, natural and cultural space”, which was held on 6-8 september 2011 in wichrowo forests (n poland, the forest division of wichrowo). due to the limited time of field work, the research focused on the most valuable and representative habitats in the study area, which include the approved or designed special areas of conservation natura 2000. lichens and lichenicolous fungi of wichrowo forests have never been investigated, and the geographical location of this forest complex and high heterogeneity of forest plant communities make the site particularly interesting in lichenological terms. study area wichrowo forests (the forest division of wichrowo) cover an area of 17645 ha and consist of two dense parts (complexes) corresponding to two subdivisions: obręb łaniewo and obręb wichrowo (fig. 1). each of them is situated in a different natural forest region: the forest complex of łaniewo in the dzielnica elbląsko-warmińska district of the baltic region, whereas the complex of wichrowo in the dzielnica mazursko-podlaska region in the pojezierze mazurskie lakeland (trampler et al. 1990). according to physical and geographical division proposed by kondracki (2001), the forest division of wichrowo is located in the area of four mesoregions: pojezierze olsztyńskie lakeland, nizina sępopolska lowland, równina ornecka plain and wzniesienia górowskie hills. the land relief of wichrowo forests is very diverse and includes a plateau associated with the vistulian (baltic) glaciation. the receding glacier left behind many moraine ramparts of different directions and a dense network of dead-ice depressions transformed into peat bogs. the highest elevation in this area reaches 172 m a.s.l. (kraszewo wilderness), the lowest one – 36 m a.s.l. (łaniewo village). the the preservation status of the lichen biota 137 © the author(s) 2014 published by polish botanical society forest division area is cut through by several rivers flowing in eroded channels, which in some places developed as gorges with a relative altitude above 60 m, and this gives the landscape typical mountain features. wichrowo forests come mostly from afforestation and planting (73.1%), and to a small extent – from natural regeneration (2.61%) or regrowth (3.6%) (plan urządzenia 2009). scots pine (pinus sylvestris) is the dominant species and it covers 60.3% of the forest division, whereas deciduous species cover only 27.6%. the paper presents the results of the research conducted in the designed special area of conservation (sac) natura 2000 „middle łyna river valley – smolajny”. the idea of creating sac in wichrowo forests came up as a result of nature inventory carried out in the state forests in 2006 and 2007. the area of „middle łyna river valley – smolajny” sac includes 2953.7 ha with the łyna valley as a central axis (ca. 30 km of the river gap section between the town of dobre miasto and lidzbark warmiński) together with the estuary section of the kirsna river flowing into łyna (pakulnicka et al. 2009). forest cover 79% of the area and they are abundant in plant communities listed in the habitat directive, including riparian black alder forest, ash-alder forest, as well as subcontinental and hillside lime-oak-hornbeam forests. the remaining part are open areas with extensively used lowland meadows and many oxbow lakes in different developmental stages. fig. 1. location of the analysed area in the wichrowo forest (the forest division of wichrowo): 1 – forest areas, 2 – boundary of the designed special area of conservation natura 2000 „middle łyna river valley – smolajny” 138 d. kubiak et al. © the author(s) 2014 published by polish botanical society material and methods the search for species was conducted in september 2011 in two working groups at a total of 30 research sites (tab. 1). at most sites, a complete inventory of species was made, including all ecological groups of lichens. in few cases, only rare or otherwise interesting species were found. species identified in the field were listed without collection of herbarium specimens. in other cases – specimens were collected for further detailed taxonomic (anatomical or biochemical) analysis in a laboratory. the collected herbarium material was deposited in the following herbaria: oltc, sltc and gpn (gorce national park). the species nomenclature follows fałtynowicz (2003) and kirk (2013), except the genus varicellaria (schmitt et al. 2012). threat categories of lichens were quoted after cieśliński et al. (2006), and the list of lichensindicators of lowland old-growth forests − after czyżewska and cieśliński (2003). results altogether 159 species were found in the study area, including 151 lichen species (lichenized fungi) and 8 species of saprotrophic or parasitic (lichenicolous) fungi (tab. 2). epiphytes dominate in the biota of lichens. most of the species were found on oak (79 species), hornbeam (71), maple (60) and linden trees (44). the largest number of species excluded for a given phorophyte was recorded for hornbeam, oak and maple – 12 taxa each. other ecological groups were represented by much smaller numbers: epixylic lichens – 22 species (including 12 growing exclusively on wood) and epigeic species – only 4 species. the identified lichen biota is represented by 19 taxa legally protected in poland, including 18 strictly protected species and 1 partially protected species. threatened lichens (facing a risk of extinction in poland) are represented by 57 species (tab. 2). they represent the following threat categories: critically endangered (cr) – 3 species, endangered (en, high risk of extinction) – 12, vulnerable (vu) – 21, least concerned (lc) – 3, near threatened (nt) – 16, data deficient (dd) – 2. the biota of lichens in the study area is represented by many species that have been previously known from only very few sites in the country, and their distribution, habitat preferences and potential threats are not well researched. the group includes: bacidia hemipolia f. pallida, biatora chrysantha, b. pontica, biatoridium monasteriense, fellhaneropsis vezdae, and lecanora farinaria. discussion the number of lichen species reported from the study area is considered to be relatively high. it is higher than the total number of species recorded in important but small refugia of forest lichens in the southern part of the pojezierze olsztyńskie lakeland, including the nature reserves of „dęby napiwodzkie” (100 species) and koniuszanka ii” (80 species) (kubiak 2011), but smaller than the number of species reported from the much larger forest nature reserve „las warmiński” – ca. 220 the preservation status of the lichen biota 139 © the author(s) 2014 published by polish botanical society table 1 major characteristics of the studied sites no. location (forest section; geographical coordinates) forest stand type (dominated tree/age) 1 403f; 54°02’04.2’’n/20°23’36.9’’e oak-hornbeam forest (q 92) 2 346l; 54°02’44.5’’n/20°24’29.0’’e old-growth oak forest 3 310h; 54°03’05.7’’n/20°26’18.0’’e lime-oak-hornbeam forest (q 137) 4 366b; 54°02’41.7’’/20°24’25.7’’e lime-oak-hornbeam forest (q 192) 5 366b; 54°02’36.4’’n/20°24’15.1’’e lime-oak-hornbeam forest (q 192) 6 347i; 54°02’44.2’’n/20°24’24.3’’e group of old maple trees 7 346h; 54°02’478’’n/20°24’35.5’’e group of trees at the crossroads 8 346l; 54°02’45.7’’n/20°24’45.9’’e lime-oak-hornbeam old-growth forest, hillside lime-oak-hornbeam forests (q 242) 9 291f; 54°03’40.0’’n/20°27’00.2’’e oak forest (q 132) 10 342c (/343a); 54°03’03.5’’n/20°26’07.2’’e lime-oak-hornbeam forest (q 127) 11 343f; 54°02’57.9’’/20°26’02.3’’e elm-ash forest (a 52) 12 314j; 54°03’09.6’’n/20°25’34.6’’e mixed pine-oak forest 13 344d; 54°02’57.8’’n/20°25’32.3’’e degenerated oak-hornbeam forest (dominance of hornbeam) 14 363m; 54°02’30.6’’n/20°25’16.2’’e lime-hornbeam forest (t 75) 15 363o; 54°02’30.4’’n/20°25’06.8’’e alder forest (a 62) 16 363k; 54°02’30.6’’n/20°25’10.2’’e spruce forest (pa 21) 17 363j; 54°02’37.0’’n/20°25’00.5’’e oak-hornbeam forest (q102) 18 589c; 53°59’43.5’’n/20°26’40.0’’e lime-oak-hornbeam forest (q107) 19 560g; 53°59’’59.8’’n/20°26’’22.3’’e spruce managed forest (q 52) 20 561a; 54°00’’11.0’’n/20°26’’01.9’’e mixed spruce-oak forest (pa 97) 21 557h; 54°00’16’’n/20°25’43.5’’e lime-oak-hornbeam forest (q 42) 22 528f; 54°00’255’’n/20°25’187’’e mixed spruce-pine-oak forest (ps 127) 23 560k; 53°59’57’’n/20°26’215’’e lime-oak-hornbeam forest (q 107) 24 589a; 53°59’44’’n/20°26’38’’e spruce-pine forest 25 560j; 53°59’589’’n/20°26’255’’e riparian black alder forest 26 197b; 54°05’59.4’’n/20°29’53.9’’e black alder forest in deep stream valley, surrounded by pine forest (ps 72) 27 196g; 54°06’11.1’’n/20°29’57.6’’e mixed oak-pine forest (ps 102) 28 185c; 54°06’47.5’’n/20°30’51.6’’e ash-alder forest 29 179k; 54°06’54.9’’n/20°31’24.0’’e oak forest (q 99) 30 309h; 54°03’18.5’’n/20°26’34.0’’e lime-oak-hornbeam forest abbreviations: a – alnus, ps – pinus, pa – picea q – quercus, t – tilia table 2 list of taxa recorded in the designed special area of conservation natura 2000 „middle łyna river valley – smolajny” species localities substrat status of the species absconditella lignicola vězda & pišút 14 epx acrocordia gemmata (ach.) a. massal. 8, 11, 17, 18, 30 cb, q, pt, ug vu agonimia allobata (stizenb.) p. james 1 q amandinea punctata (hoffm.) coppins & scheid. 2, 4 ap, cb, q anisomeridium polypori (ellis & everh.) m.e. barr 1, 2, 8, 11, 21, 26 ap, pt, q, s, ug alyxoria varia (pers.) ertz & tehler 4, 8 ap nt arthonia byssacea (weigel) almq. 28 fe en a. didyma körb. 4 cb en 140 d. kubiak et al. © the author(s) 2014 published by polish botanical society a. mediella nyl. 1 q vu a. radiata (pers.) ach. 1, 3, 9, 10, 14, 27, 30 cb, q, sa a. spadicea leight. 1, 3, 8, 10, 11, 18, 30 ap, ag, cb, q a. vinosa leight. 1, 11 ag, cb, q nt arthothelium ruanum (a. massal.) körb. 4, 5, 8, 11, 28 cb, fe, tc nt *arthrorhaphis aeruginosa r. sant. & tønsberg 25 cladonia coniocraea/ ag bacidia arceutina (ach.) rehm. & arnold 4, 16, 21, 23, 30 cb, pt en b. fraxinea lönnr. 2, 21, 30 cb, pt dd b. hemipolia f. pallida czarnota & coppins 1, 9, 20 q b. rubella (hoffm.) a. massal. 4, 9, 28, 30 ap, fe, pt vu b. subincompta (nyl.) arnold 1, 2, 4-6, 17, 18 ap, cb, q en *bacidina chloroticula (nyl.) vězda & poelt 6 dead thallus of parmelia sulcata/ap b. phacodes (körb.) vězda 16 pt b. sulphurella (samp.) m. hauck & v. wirth 1, 3, 4, 6, 10, 11, 26 ag, ap, cb, pa, q, epx biatora chrysantha (zahlbr.) printzen 12 q b. efflorescens (hedl.) räsänen 1-6, 8, 18, 27, 29, 30 ap, cb, q, tc vu b. globulosa (flörke) fr. 1, 8, 12, 28 ap, fe, q b. ocelliformis (nyl.) arnold 3 cb vu b. pontica printzen & tønsberg 5 tc biatoridium monasteriense j. lahm. ex körb. 11 ug nt buellia griseovirens (turner & borrer ex sm.) almb. 1-4, 6, 8-11, 16, 30 ap, cb, q, tc calicium adspersum pers. 2, 4 q en c. salicinum pers. 2, 4 q vu c. viride pers. 2, 4, 5, 7 ap, q vu candelariella efflorescens r.c. harris & w.r. buck 1 q c. xanthostigma (pers ex ach.) lettau 4 ap catillaria nigroclavata (nyl.) j. steiner 16 pt chaenotheca chlorella (ach.) müll. arg. 28 fe cr ch. chrysocephala (ach.) th.fr. 1-3, 5, 7-9, 27, 28, 29 ap, ps, q ch. ferruginea (turner ex sm.) mig. 1-5, 8, 9, 11,12, 16, 28, 29 ap, cb, pa, ps, q ch. furfuracea (l.) tibell 11, 21 ag, pa nt ch. stemonea (ach.) müll. arg. 2-4, 29 pa, q en ch. trichialis (ach.) hellb. 2-5, 26, 28, 29 ag, ap, pa, q nt *chaenothecopsis savonica (räsänen) tibell 4 cb (exposed dead wood) chrysothrix candelaris (l.) j.r. laundon 4, 5, 7, 8, 28, 29 fe, q sp, cr cladonia chlorophaea (flörke ex sommerf.) spreng. 29 q c. coniocraea (flörke) spreng. 1-5, 6, 9-12, 16, 26, 27, 29 ag, bp, cb, pa, ps, q, tc, epx, ter c. digitata (l.) hoffm. 4, 11, 27 ps, epx c. fimbriata (l.) fr. 1, 9, 16, 26 ps, q, tc, s c. furcata (huds.) schrad. 16 ter cliostomum corrugatum (ach.) fr. 5 q cr c. griffithi (sm.) coppins 1 q vu *clypeococcum hypocenomycis d. hawksw. 1, 2, 27 hypocenomyce scalaris/pa, ps, q coenogonium pineti (schrad.) lücking & lumbsch 1-6, 9, 10, 12, 16, 26, 27, 29, 30 ag, ap, cb; pa, ps, q, tc, epx table 2 – cont. the preservation status of the lichen biota 141 © the author(s) 2014 published by polish botanical society evernia prunastri (l.) ach. 1-10, 12,16, 28, 29, 30 ap, cb, ee, fe, pav, pas, p, q, tc, epx pp, nt fellhanera gyrophorica sérus., coppins, diederich & scheid. 1, 2, 9, 10, 18, 20, 29, 30 cb, q lc fellhaneropsis myrtillicola (erichsen) sérus. & coppins 20 pa f. vezdae (coppins & p. james) sérus. & coppins 3 cb lc flavoparmelia caperata (l.) hale 8 q sp, en fuscidea arboricola coppins & tønsberg 1-3, 15, 25 ag, cb, q f. pusilla tønsberg 1, 2, 15, 25, 27 ag, cb, ps, q, tc, epx graphis scripta (l.) ach. s.l. 1-3, 5, 6, 8-11, 30 ap, cb, ca, tc nt hypocenomyce scalaris (ach. ex lijl.) m. choisy 1, 2, 4, 5, 7, 9, 12, 27 cb, pa, ps, q sp., tc hypogymnia physodes (l.) nyl. 1-9, 11, 12, 15, 16, 26, 27, 29, 30 ag, ap, cb, ee, pa, ps, p, q, tc, s, sa, epx *ilosporiopsis christiansenii (b. l. brady & d. hawksw.) d. hawksw. 6 physcia adscendens/ ap imshaugia aleurites (ach.) s.l.f. mey. 7, 9 ps, q sp lecania cyrtella (ach.) th. fr. 6 ap l. naegelii (hepp) diederich & van den boom 6 ap lecanora albella (pers.) ach. 4 ap en l. argentata (ach.) malme 4-6, 10, 30 ap, cb, q l. carpinea (l.) vain. 1, 2, 5, 8, 16, 28, 30 cb, fe, pt, q l. chlarotera nyl. 1-5, 8-10, 16, 28, 30 ap, cb, p, q, tc l. compallens herk & aptroot 7, 11, 16 ag, cb, fe, p, q, tc l. conizaeoides nyl. ex cromb. 1, 2, 9 cb, pa, ps, tc l. expallens ach. 1-11, 14-16, 26, 27, 28, 29 ap, ag, cb, ca, fe, pa, q, ug, sa, tc l. farinaria borrer 1 cb l. glabrata (ach.) malme 5, 8, 30 cb l. pulicaris (pers.) ach. 1, 2, 4, 5, 9, 15, 16 ag, cb, q, tc l. rugosella zahlbr. 16, 30 q, pt l. sarcopidoides (a. massal.) hedl. 24 ps nt l. thysanophora r.c. harris 2, 3, 5, 10, 13, 30 cb, q lecidea nylanderii (anzi) th.fr. 2 cb lecidella elaeochroma (ach.) m. choisy 4-6, 8-10, 16, 26, 28, 30 ap, cb, fe, q, pt, s l. flavosoreditata (vězda) hertel & leuckert 4 ap l. subviridis tønsberg 2, 4, 5, 16, 17, 18 ag, cb, tc lepraria elobata tønsberg 1-6, 8, 9,12, 13, 16, 26, 27 ag, cb, q, ps, tc l. incana (l.) ach. 1-14, 16, 26, 27, 28, 29 ap, ag, bp, cb, fe, pa, ps, q, tc l. jackii tønsberg 1, 2, 27 cb, pa, ps, tc l. lobificans nyl. 1-13, 26, 28, 29, 30 ap, bp, cb, fe, q, ug, tc, pt, s l. rigidula (b. de lesd.) tønsberg 1, 6, 26, 28 ag, ap, fe, tc l. vouauxii (hue) r.c. harris 9 ap lichenomphalia umbellifera (l.) redhead et al. 19 epx nt melanohalea exasperatula (nyl.) o. blanco et al. 2, 6 ap sp melanelixia glabratula (lamy) sandler & arup 1-11, 28, 30 ap, cb, pav, q, tc, pt sp m. subaurifera (nyl.) o. blanco et al. 8 pav sp micarea byssacea (th. fr.) czarnota, guz.-krzemiń. & coppins 14, 23 pt, tc m. melaena (nyl.) hedl. 15 epk nt table 2 – cont. 142 d. kubiak et al. © the author(s) 2014 published by polish botanical society m. micrococca (körb.) gams ex coppins 4, 8, 15, 16, 21 fe, ps-st c, pt, q, epx m. misella (nyl.) hedl. 22 epx m. nitschkeana (j. lahm. ex rabenh.) harm. 16 pt m. viridileprosa coppins & van den boom 22 ps (*)microcalicium dissemnatum (ach.) vain. 4 q *monodictis epilepraria kukwa & diederich 1, 8 lepraria sp./q and ap mycobilimbia epixanthoides (nyl.) vitik., ahti, kuusinen, lommi & t. ulvinen 1, 6, 9, 10, 17, 18, 30 ap, cb, q *mycocalicium subtile (pers.) szatala 8 q (exposed dead wood) *nectriopsis rubefaciens (ellis & everh.) m.s. cole & d. hawksw. 15 parmelia sulcata/ wood ochrolechia bahusiensis h. magn. 2, 6-10 ap, cb, q, tc vu1 o. microstictoides räsänen 2, 5, 8, 30 ap, cb, q, tc o. turneri (sm.) hasselrot 4, 7-9 ap, epx opegrapha rufescens pers. 14 tc vu o. vermicellifera (kunze) j.r. laundon 5, 8 cb, tc en o. vulgata (ach.) ach. 18 cb vu pachyphiale fagicola (arnold) zwackh 26, 28 ap, s vu parmelia saxatilis (l.) ach. 4, 6, 8 ap, q sp p. sulcata taylor 1-4, 6, 8, 9, 11, 16, 26, 27, 28, 30 ag, ap, cb, fe, q, s, tc parmeliopsis ambiqua (wulfen) nyl. 9 ps sp peltigera didactyla (with.) j.r. laudon 12 ter sp p. hymenina (ach.) delise 9 epx sp, dd p. praetextata (flörke ex sommerf.) zopf 8, 17, 18 ap, q, epx, ter sp, vu pertusaria albescens (huds.) m. choisy & werner 3-6, 9, 30 ap, cb, q, tc p. amara (ach.) nyl. 1-3, 5, 7-10, 12, 13, 18, 27, 28, 29 ap, cb, fe, q p. coccodes (ach.) nyl. 1-4, 6, 8-10, 12, 13, 16, 30 ap, cb, q, tc nt p. coronata (ach.) th.fr. 4, 5, 8, 16 cb, q, tc vu p. flavida (dc.) j.r. laundon 4 cb en p. leioplaca dc. 3-5, 8-10, 14, 30 ap, cb, tc nt p. pertusa (l.) tuck. 3, 10 cb vu p. pupillaris (nyl.) th.fr. 18 cb nt phlyctis argena (ach.) flot. 1-9, 11-13, 16, 18, 26, 27, 28, 30 ag, ap, cb, ca, fe, p, pt, q, s, tc physcia adscendens (fr.) h. olivier 6 ap p. tenella (scop.) dc. 4, 6 ap, q physconia enteroxantha (nyl.) poelt 1, 4, 6, 9 ap, q p. grisea (lam.) poelt ap placynthiella dasaea (stirt.) tønsberg 2, 3 epx p. icmalea (ach.) coppins & p. james 2, 7, 15 epx platismatia glauca (l.) w.l. culb. & c.f culb. 2, 3, 5-9, 11, 26, 27, 29 ag, ap, cb, pa, q, tc sp porina aenea (wallr.) zahlbr. 1-5, 8, 10, 11, 16, 30 cb, tc, q pseudevernia furfuracea (l.) zopf 9, 29 ee, ps sp psilolechia clavulifera (nyl.) coppins 12 pa (exposed roots) nt pycnora sorophora (vain.) hafellner 9 ps pyrenula nitida (weigel) ach. 5, 8 cb vu p. nitidella (flörke ex schaer.) müll. arg. 5, 8, 10 cb, fe en table 2 – cont. the preservation status of the lichen biota 143 © the author(s) 2014 published by polish botanical society species (kubiak 2012). so far pine forest habitats with the sites of terricolous lichens and non-forest habitats – e.g., roadside woodlots, have been investigated to a small extent only. furthermore, the data on lichenicolous fungi should be regarded as a contribution to the knowledge about the group. it appears that the species diversity of lichens in the most valuable fragments of the study area, including mesoand eutrophic communities of oak-hornbeam forests, riverine forests and alder swamp woods, is well evidenced by the conducted research. mixed lime-oak-hornbeam forests are one of the most valuable plant communities in terms of lichen biota heterogeneity. many stenotopic lichens occur at these sites, in particular epiphytes and epixylites (cieśliński et al. 1995; czyżewska 2003). although there are relatively many thematically diverse papers on lime-oakhornbeam forests, the contribution of lichens in these communities is not thoroughly explored and lichens are basically overlooked in phytosociological studies. biodiversity of forest ecosystems largely depends on their natural conditions (jaroszewicz 2007) and in the case of organisms like lichens, this is a determining factor (cieśliński, tobolewski 1988; cieśliński et al. 1996; czyżewska, cieśliński 2003; kubiak, sucharzewska 2012). species that are natural components of forest biocenoses dominate in the lichen biota of the study area. stenotopic lichens are well ramalina farinacea (l.) ach. 1-3, 5, 6, 8-10, 12, 16, 28, 29, 30 ap, cb, fe, p, q, tc, pt sp, vu r. pollinaria (westr.) ach. 2, 4 ap, cb, q, tc sp, vu reichlingia leopoldii diederich & scheid. 3, 11, 12 ag, cb, q, ug rinodina degeliana coppins 1 q r. efflorescens malme 1, 2, 4, 6 ap, cb, q, tc ropalospora viridis (tønsberg) tønsberg 2, 3, 5, 8-13, 21, 23, 26, 30 ag, cb, fe, pt, q, tc scoliciosporum chlorococcum (graewe ex stenh.) vězda 2, 8 cb, epx s. sarothamni (vain.) vězda 2, 4, 6, 15 ag, ap, tc *taeniolella punctata m.s. christ. & d. hawksw. 8 graphis scripta/cb thelocarpon epibolum nyl. 15 epx lc trapeliopsis flexuosa (fr.) coppins & p. james 3, 7 epx t. granulosa (hoffm.) lumbsch 2, 4, 9 epx usnea filipendula stirt. 7, 29 q sp, vu u. subfloridana stirt. 9, 29 cb, q sp, en varicellaria hemisphaerica (florke) schmitt & lumbsch 3, 5, 6, 7, 10 ap, q, tc vu violella fucata (stirt.) t. sprib. 1, 2, 4-6, 10, 15, 27 ap, ag, cb, tc, sa vulpicida pinastri (scop.) j.-e. mattsson 15 ag sp, nt xanthoria parietina (l.) beltr. 6 ap x. polycarpa (hoffm.) rieber 6 ap zwackhia viridis (ach.) poetsch & schied. 2-6, 8-11 ap, ag, cb, q vu abbreviations: * – saprotrophic or parasitic (lichenicolous) fungus; ag – alnus glutinosa, ap – acer platanoides, bp – betula pendula, ca – corylus avellana, cb –carpinus betulus, ee – euonymus europaea, fe – fraxinus excelsior, pa – picea abies, pav – padus avium, ps – pinus sylvestris, p – populus sp., pt – populus tremula, q – quercus sp., s – salix sp., sa – sorbus auccuparia, tc – tilia cordata; ter – terricolous, epx – epixylic; sp – strictly protected species, pp – partially protected species; cr – critically endangered, en – endangered, vu – vulnerable, nt – near threatened, lc – least concern, dd – data deficient; 1 – as o. androgyna s.l. table 2 – cont. 144 d. kubiak et al. © the author(s) 2014 published by polish botanical society represented − they are less and less frequent outside the area of extensive and well preserved forests. when comparing the species composition of lichens recorded in the łyna valley with the results obtained in the strongly deforested, agricultural area of the równina warmińska plain (szymczyk, zalewska 2008), the number of lichen species growing in wichrowo forests is much larger (równina warmińska plain – 96 species, the łyna valley – 152). there were only 44 lichen species common for both compared areas – mostly widespread, ubiquitous and hemerophilous. although the number of protected species recorded in the równina warmińska plain is similar – 14 species (the łyna valley – 19), threatened species were much fewer – 18 species (the łyna valley – 57). the relatively large number of lichens attached to the bark of old oak trees, rare or even very seldom species, threatened with extinction in poland, is one of the most valuable components of the lichen biota, and as it appears – differential for the studied fragment of wichrowo forests. the group includes: calicium adspersum, c. viride, chrysothrix candelaris, cliostomum corrugatum i microcalicium disseminatum. in poland, these taxa are regarded as indicators of lowland old-growth forests (czyżewska, cieśliński 2003). a total of 21 species with this status were found in the study area (tab. 2). this number is relatively large compared to previously researched and described, similar refugia of forest lichens (czyżewska, cieśliński 2003). some of the listed species occur in the study area with a relatively high frequency (eg., chrysothrix candelaris, fellhanera gyrophorica, varicellaria hemisphaerica, zwackhia viridis) compared to forests outside the nature reserves. the occurrence of many lichen species in forests is determined by high species and age diversity of forest stands, owing to which individual taxa make use of the evolutionary consolidated ecological amplitude in relation to substrate (tree species), and consequently, may exist at a given site (see cieśliński 2008). oak is of particular significance in this respect, because of the longevity and diverse structure of periderm varying with age (cieśliński 2008; kubiak, sucharzewska 2012). a total of 79 species of lichens and saprotrophic fungi were found on the bark of this phorophyte in the study area, which is a relatively large number (kubiak 2011) and is higher than the number of species recorded on the bark of oak trees in the forest reserves „dęby napiwodzkie” and „koniuszanka ii” (71 species) with unique oldgrowth oak forest (kubiak 2011). many authors emphasize the importance of old oak trees for the preservation of species richness epiphytic lichens in forests, both in quantitative and qualitative terms (cieśliński 2008; johansson et al. 2009; kubiak, zalewska 2009; kubiak, sucharzewska 2012). summary as evidenced by the results presented and compared to other forests in warmia and masuria not included yet in the nature reserve conservation programme, the analysed fragment of wichrowo forests is characterised by above-average lichenological values and deserve protection. due to the unique geographical location and land relief, the area functions as an ecological corridor and is potentially very valuable in the nature conservation system of the warmia-masuria province. although the preservation status of the lichen biota 145 © the author(s) 2014 published by polish botanical society after verification, the area was ultimately removed from the list of potential sites of community importance (sci), its natural values are high and fully deserve detailed exploration and protection. due to the limited scope of the research conducted, further lichenological research in wichrowo forests should be continued, both in other types of forest plant communities and in non-forest habitats directly connected with a forest. acknowledgements. the authors are very grateful to mr. andrzej reguła (the forest division manager) and other employees of the wichrowo forest inspectorate for their comprehensive assistance during the field work, and to the anonymous reviewers for helpful remarks on the manuscript. references cieśliński s. 2003. atlas rozmieszczenia porostów (lichenes) w polsce północno-wschodniej. phytocoenosis (n.s.) 15, suppl. cartogr. geobot. 15: 1-430. cieśliński s. 2008. znaczenie ochrony rezerwatowej dla zachowania bioty porostów (ascomycota lichenisati) w puszczy kozienickiej. stud. i mat. cepl, rogów 10, 3 (19): 99-109. cieśliński s., czyżewska k. 1992. problemy zagrożenia porostów w polsce. wiad. bot. 36 (1–2): 5-17. cieśliński s., czyżewska k. 2003. czerwona lista porostów wymarłych i zagrożonych w polsce. monogr. bot. 91: 13-49. cieśliński s., czyżewska k., fabiszewski j. 2006. red list of the lichens in poland. (in:) z. mirek, k. za-zarzycki, w. wojewoda, z. szeląg (eds). red list of plants and fungi in poland. w. szafer institute of botany, polish academy of sciences, kraków: 71-89. cieśliński s., czyżewska k., faliński j. b., klama h., mułenko w., żarnowiec j. 1996. relicts of the primeval (virgin) forest. relict phenomena. (in:) j.b. faliński, w. mułenko (eds). cryptogamous plants in the forest communities of białowieża national park (project crypto 3). phytocoenosis 8 (n.s.), archivum geobot. 6: 197-216. cieśliński s., czyżewska k., glanc k. 1995. lichenes. (in:) j.b. faliński, w. mułenko (eds). cryptogamous plants in the forests communities of białowieża national park. general problems and taxonomic groups analysis (project crypto). phytocoenosis 7 (n.s.), archiv. geobot. 4: 75-86. cieśliński s., fałtynowicz w. 1993. note from editors. (in:) s. cieśliński, w. fałtynowicz (eds). atlas of the geographical distribution of lichens in poland 1: 7-8. w. szafer institute of botany, polish academy of sciences, kraków. cieśliński s., tobolewski z. 1988. porosty (lichenes) puszczy białowieskiej i jej zachodniego przedpola. phytocoenosis 1 (n.s.), suppl., cartograph. geobot. 1: 3-216. czyżewska k. 2003. ocena zagrożenia bioty porostów polski. monogr. bot. 91: 241-249. czyżewska k., cieśliński s. 2003. porosty – wskaźniki niżowych lasów puszczańskich w polsce. monogr. bot. 91: 223-239. fałtynowicz w. 2003. the lichens, lichenicolous and allied fungi of poland. an annotated checklist. w. szafer institute of botany, polish academy of sciences, kraków. jaroszewicz b. 2007. różnorodność biologiczna lasów polskich. wszechświat 108 (46): 216-221. johansson v., bergman k., o., lättman h., milberg p. 2009. tree and site quality references of six epiphytic lichens growing on oaks in southeastern sweden. ann. bot. fenn. 46: 496-506. kirk p.m. 2013. species fungorum. digital resources at www.speciesfungorum.org/names/names.asp. accessed 15 april 2013. kondracki j. 2001. geografia regionalna polski. pwn, warszawa. kubiak d. 2011. stan zachowania bioty porostów w rezerwatach „dęby napiwodzkie” i „koniuszanka ii” na pojezierzu olsztyńskim. parki nar. rez. przyr. 30 (3–4): 27-39. kubiak d. 2012. protected and threatened lichen species of the nidzica primeval forest (n poland). (in:) l. lipnicki (ed.). lichen protection – protected lichen species. . sonar literacki, gorzów wlkp.: 263-276. kubiak d., sucharzewska e. 2012. porosty – wskaźniki niżowych lasów puszczańskich w zespołach leśnych rezerwatu „las warmiński” (nadleśnictwo nowe ramuki). sylwan 156 (8): 627-636. 146 d. kubiak et al. © the author(s) 2014 published by polish botanical society kubiak d., zalewska a. 2009. notes on caloplaca lucifuga (teloschistales, ascomycota) in poland. acta mycol. 44 (2): 239-248. pakulnicka j., górski a., lewandowski k., kruszelnicki j. 2009. dolina środkowej łyny – smolajny. (in:) w: hołdyński c., krupa m. (eds). obszary natura 2000 w województwie warmińsko-mazurskim: 137-140. wydawnictwo mantis, olsztyn. plan urządzenia lasu na lata 2009-2018. nadleśnictwo wichrowo, obręb wichrowo. program ochrony przyrody. stan na 1.01.2009 r. biuro urządzania lasu i geodezji leśnej oddział w olsztynie. schmitt i., otte j., parnmen s., sadowska-deś a.d., lucking r., lumbsch h.t. 2012. a new circumscription of the genus varicellaria (pertusariales, ascomycota). mycokeys 4: 23-36. szymczyk r., zalewska a. lichens in the rural landscape of the warmia plain. acta mycol. 43 (2): 215230. trampler t., kliczkowska a., dmytreko e., sierpińska a. 1990. regionalizacja przyrodniczo-leśna na podstawach ekologiczno-fizjograficznych. pwril, warszawa. ustawa z dnia 28 września 1991 r. o lasach dz.u. 1991 nr 101 poz. 444 (z późniejszymi nowelizacjami). ustawa z dnia 16 kwietnia 2004 r. o ochronie przyrody dz.u. 2004 nr 92 poz. 880. stan zachowania bioty porostów projektowanego specjalnego obszaru ochrony siedlisk natura 2000 „dolina środkowej łyny – smolajny” streszczenie praca przedstawia wyniki badań lichenologicznych przeprowadzonych w dniach 6-8 września 2011 r. w ramach sesji terenowej towarzyszącej 25 zjazdowi lichenologów polskich, który odbywał się w medynach koło lidzbarka warmińskiego. celem sesji było poznanie zasobów gatunkowych porostów położonych na terenie lasów wichrowskich (nadleśnictwo wichrowo), istniejących lub projektowanych, specjalnych obszarów ochrony siedlisk (soos) natura 2000. w tej pracy przedstawiono wyniki inwentaryzacji przeprowadzonej na obszarze zaproponowanego do utworzenia soos „dolina środkowej łyny – smolajny”. obszar o powierzchni 2953 ha obejmuje głównie zbiorowiska leśne, związane z przełomowym odcinkiem rzeki łyny (grądy typowe, grądy zboczowe, łęgi, olsy). na analizowanym terenie znaleziono 159 taksonów, w tym 151 gatunków porostów (grzybów zlichenizowanych) oraz 8 gatunków grzybów saprotroficznych lub pasożytniczych (naporostowych). biota ta obejmuje 19 taksonów objętych w polsce ochroną oraz 57 zagrożonych w skali kraju wymarciem. ponadto, stwierdzono gatunki znane dotychczas w kraju z bardzo nielicznych stanowisk, do których można zaliczyć: bacidia hemipolia f. pallida, biatora chrysantha, b. pontica, biatoridium monasteriense, fellhaneropsis vezdae, lecanora farinaria. na podstawie uzyskanych wyników można stwierdzić, że analizowany fragment lasów wichrowskich, na tle innych kompleksów leśnych warmii i mazur nie objętych dotychczas ochroną rezerwatową, wykazuje ponadprzeciętne wartości lichenologiczne, które zasługują na ochronę. 2014-06-30t23:14:27+0200 polish botanical society 2014-09-03t19:46:14+0200 polish botanical society 2014-09-02t20:09:56+0200 polish botanical society 2014-11-20t23:13:07+0000 polish botanical society 2014-11-20t23:12:08+0000 polish botanical society congratulations 149 a congratulatory message to prof. maria ławrynowicz from the european mycological association the volume of acta mycologica celebrates the jubilee of prof. maria ławrynowicz, and the european mycological association is proud to contribute a message of congratulations. prof. ławrynowicz is a leading and enormously influential figure in european mycology. as a scientist, she is best known for her meticulous and highly respected work on hypogeous fungi, having contributed extensively through high-quality publications to the scientific knowledge of the taxonomy, biology and geographical distribution of these interesting and enigmatic species. for humans, who are not blessed with the ability of other mammals to detect scents, hypogeous fungi can seem secretive and notoriously difficult to find. watching prof. ławrynowicz locate their fruitbodies with apparent ease has frequently evoked admiration and amazement in equal quantities from fellow mycologists, i asked her once how she did it, and she explained, perhaps only half joking, that maybe in some previous incarnation she had been a mouse. her humour is like that: gentle and always thought-provoking. polish mycologists have always had a high profile at the congresses of european mycologists, and prof. ławrynowicz must now have attended more of these meetings then any other living mycologist, her predecessor, colleague, teacher and friend, the late prof. skirgiełło being the only other person who had a similar record of participation. years ago, in the 1980s and 1990s, these two ladies were a regular sight at the congresses, and what a formidable pair they made! poland was well represented. as prof. skirgiełło faded, it became more challenging for her to attend these events, and european mycologists will remember how she was helped and supported by prof. ławrynowicz, so much so that some younger scientists may have even got the impression that the one was the nurse of the other. by her devoted care of an elderly colleague, often to the detriment of her own participation, prof. ławrynowicz won universal admiration and respect from her european mycologist friends. 150 if i were to pick a defining moment to illustrate her influence, it would be her role at the european congress in oslo in 1985, when she, with a small group of other far-sighted mycologists, set up the european council for conservation of fungi. this was the first grouping anywhere in the world to address the need for fungi to be protected. prof. ławrynowicz was truly a pioneer in fungal conservation. the council continues to function, and inspired the establishment of similar bodies in other continents and, eventually, the foundation of the international society for fungal conservation of which, not surprisingly, she was also a founder member. that same preoccupation with infrastructure – something which has chronically been lacking in mycology – led her to become a founder member also of the european mycological association, which is today sending her congratulations. but it was also reflected in her work in poland. her influence on polish mycology is very far-reaching. as editor-in-chief, she has been a guiding influence on the very positive development of acta mycologica, the journal in which her jubilee is being celebrated, and as a patient but effective force for change, she has played a great role in the recent establishment of the polish mycological society. she is well aware, however, that infrastructure is nothing without motivated people who can use it to promote mycology, and it is that her skills as a teacher shine most brightly. over the years she has inspired many students who have subsequently gone out into the world with an understanding that fungi are important – that we live on this planet only thanks to the essential role as nature’s recyclers. some of those students have become respected and well-known mycologists in their own right. it would be unfair to name some and not others, and i won’t even try, but the fact that they exist is perhaps the greatest achievement of the lady whom we celebrate in this special volume. the european mycological association sends her congratulations, and warm wishes for many more years of productive work with the fungi. david minter president european mycological association 151 maria ławrynowicz’s jubilee hypogeous fungi specialists are rare because the study of these mycetes is difficult, indeed, their underground habit means that they are unnoticed and their discovery is often fortuitous, luckily examining herbariums can uproot a mine of information. dr. maria ławrynowicz is one of these specialists such as hawker in the united kingdom, hesse and gross in germany, knapp and schwärzel in switzerland, hollós and szemere in hungary. in 1988, she publishes her first studies on the elaphomycetales and the tuberales in the series “polish flora”. she becomes a specialist for the elaphomycetales and european tuberales, and publishes the chorological distribution respectively in 1989 and 1990. in 1992, in an article on the truffle geographical distribution in northern europe, from the analytical study of samples held in 44 herbariums, she shows the presence of hypogeous fungi, just like epigeous mushrooms and plants vary much more from the south to the north direction than west to east. the presence of tuber magnatum (italian white truffle), t. melanosporum (perigord truffle), t. brumale (winter truffle), t. aestivum (summer truffle), t. mesentericum, and a few species get fewer towards the north. of those with commercial interest and the northernmost is t. borchii which is present in the whole of europe up to the limits of the temperate zone. the hypogeous mushrooms depend for their development and geographical distribution on a number of factors such as climate, edaphic, biotic, historical etc. aspects. these factors act in unison and sometimes compensate each other. if tuber aestivum was first identified in poland at the end of the xixth century (1886), and its presence confirmed since 1970, t. mesentericum was only discovered in 1981. whilst only one species was known up to the fifties (t. aestivum), maria ławrynowicz’s research and that of others have identified 13 species of tuberales: t. aestivum vitt., t. aestivum vitt. forma uncinatum (chatin) montecchi and borelli, t. bellonae quél., t. borchii vitt., t. dryophilum vitt., t. excavatum vitt., t. ferrugineum vitt., t. fulgens quél., t. maculatum vitt., t. mesentericum vitt., t. puberulum berk. and br., t. repaeodorum tul., and t. rufum pico. truffles are much more than expensive goods reserved to the privileged, but a produce that is part of our culture and our european identity that we must preserve. since 2000 maria ławrynowicz has taken on board this mission through an active campaign for its culture. since 2008 truffle orchards with t. aestivum f. uncinatum (burgundy truffle) have been planted, and more recently with t. melanosporum. it is certain that in the near future, the prestigious perigord truffle will appear in poland. gerard chevalier the tuber aestivum/uncinatum scientific european group 152 a congratulatory message dear professor maria ławrynowicz, ukrainian mycologists from the department of mycology at the m.g. kholodny institute of botany in kiev send you warmest congratulations on the occasion of your anniversary, the 45th year of your scientific activity. we were delighted to invite and to see you here in ukraine, in crimea, during the xiv congress of european mycologists in 2003. we also keep good memories of our other meetings at various international conferences. your contribution to mycology is well known and highly appreciated. on behalf of all colleagues from our department of mycology, let us wish you good health, prosperity and further successful activity for the welfare of mycological science. with kind regards, prof. irina dudka head of the department of mycology prof. vasyl heluta principal scientist department of mycology m.g. kholodny institute of botany national academy of sciences of ukraine 153 the european council on conservation of fungi (eccf) congratulates professor maria ławrynowicz on the occasion of her 45th year of scientific activity prof. maria ławrynowicz, at present one of the leading experts in fungal conservation, was one of the initiators and founder members of the eccf since this organization was established in 1985. being from the beginning its active member, she became the eccf chair and made important contribution in development of this european network. as a well-known mycologist, she has significantly contributed to the knowledge of hypogeous and various other groups of fungi, always emphasizing rare and endangered species and necessity of their protection. on behalf of the eccf members from all european countries, we send to maria ławrynowicz our warmest anniversary greetings and wish her further great success in promoting fungal conservation. sincerely, beatrice senn-irlet eccf president vera hayova eccf secretary 154 13.8.1988 1. in nineteen hundred eighty eight there came from west and east to poland from eleven states sixteen mycologists. maria made conditions best and no comfort was missed. maria is a really good mushroom protectionist. 2. we came to speak and to discuss decline of fungal growth we went by foot and went by bus to pine and beech woods both. maria made us talkative on that red data list. maria is an excellent mushroom protectionist. 3. tv and press told what we did concluded, wrote and sealed, to have, five hundred years ahead; still mushrooms in the field. to have bolets and hygrocybes and truffles and bovists maria did the best for us mushroom protectionists. 4. we worked in an excellent wood, we did not fail to sing, but mainly serious work was done, which some results will bring. maria and her charming staff should thousanddfold be kissed thank you, maria, wonderful mushroom protectionist. hanns kreisel 155 first meeting of the european committee for protection of fungi, august 11-13.1988 university of lodz meeting participants. front row (from right to left): eef arnolds (wijster) – president of ecpf, maria ławrynowicz (łódź), anna elise jansen (wageningen), kurt wöldecke (hannover), wanda lasota (łódź), alina skirgiełło (warszawa), kazi-kazimierz mamos (łódź), jolanta adamczyk (łódź). back row (from right to left): johannes a. schmitt (saarbrücken), andré fraiture (meise), heikki kotiranta (oulu), bruce ing (chester), yves-l. delamadeleine (neuchâtel), hanns kreisel (greifswald), dieter benkert (berlin), jean keller (neuchâtel), rostislav fellner (praha), johan nitare (uppsala), edward tranda (łódź). photo: wojciech maliński note. the european committee for protection of fungi (ecpf) had been created in 1985 during the 9th congress of european mycologists in oslo, and renamed as the european council on the conservation of fungi (eccf) in 1989 during the 10th congress of european mycologists in tallin. 2014-09-08t17:36:52+0200 polish botanical society 2014-09-03t19:47:47+0200 polish botanical society 2014-11-20t23:12:02+0000 polish botanical society 2014-11-20t23:11:41+0000 polish botanical society 2014-11-20t23:12:35+0000 polish botanical society 2014-11-20t23:09:22+0000 polish botanical society 2014-11-20t23:13:49+0000 polish botanical society 2014-11-20t23:10:12+0000 polish botanical society 2014-11-20t23:10:35+0000 polish botanical society 2014-11-20t23:13:22+0000 polish botanical society 2014-09-06t20:22:31+0200 polish botanical society 2014-09-08t11:11:09+0200 polish botanical society 2014-09-06t20:25:31+0200 polish botanical society characteristic of tuber spp. localities in natural stands with emphasis on plant species composition 267this is an open access article distributed under the terms of the creative commons attribution 3.0 license (creativecommons.org/licenses/by/3.0/), which permits redistribution, commercial and non-commercial, provided that the article is properly cited. © the author(s) 2014 published by polish botanical society acta mycologica introduction truffles are prized fungi due to their taste and aroma. they are hypogeous and belong to pezizales, a large group of ectomycorrhizal fungi growing in symbiosis with the roots of several vascular plant species (angiosperms and gymnosperms). among the different original research paper acta mycol 49(2):267–277 doi: 10.5586/am.2014.024 received: 2014-10-22 accepted: 2014-12-05 published electronically: 2014-12-31 characteristic of tuber spp. localities in natural stands with emphasis on plant species composition dorota hilszczańska1*, aleksandra rosa-gruszecka2, hanna szmidla2 1 department of forest ecology, forest research institute, braci leśnej 3, sękocin stary, 05-090 raszyn, poland 2 department of forest protection, forest research institute, braci leśnej 3, sękocin stary, 05-090 raszyn, poland abstract fungi belonging to the genus tuber establish ectomycorrhizal symbioses with shrubs, trees and some herbaceous plants. some tuber species, for example, t. melanosporum, t. magnatum, t. aestivum are economically important because they produce edible fruiting bodies with a distinctive taste and flavor. our concept of truffle ecophysiology is dominated by the symbiosis with deciduous hosts, such as: quercus spp., fagus sylvatica, castanea sativa, corylus spp., carpinus betulus, ostrya carpinifolia, betula verrucosa, and tilia spp., whereas the real range of hosts in nature seems to be much wider. moreover, interactions between tuber mycelium and plant community could be more complex than just forming the ectomycorrhizal symbiosis. here we show our inventory of plants and soils at six truffle’ sites in the southern part of poland (nida basin and przedbórz upland). the aim of this study was to widen our understanding of ecological factors affecting tuber spp., in the context of pioneering stage of research on truffles in poland. we hope our findings will have a practical application and will help to choose suitable soils for truffle orchards. keywords: hypogeous fungi; truffles; forest stands, plant community; soil parameters * corresponding author. email: d.hilszczanska@ibles.waw.pl handling editor: maria rudawska http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.2014.024 mailto:d.hilszczanska%40ibles.waw.pl?subject=am.2014.024 268© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands species of truffles, tuber magnatum (white truffle), tuber melanosporum (black truffle), tuber aestivum (summer truffle) and tuber macrosporum vittad. (the smooth black truffle) are most valued and expensive. the first two species mentioned earlier are mainly confined to mediterranean and some south europe countries (bulgaria and serbia) while t. aestivum is widely distributed throughout europe [1,2]. the fungus forms ectomycorrhizal symbioses with many different species, including corylus avellana, quercus robur, fagus sylvatica, tilia cordata and pinus nigra. this truffle species prefers calcareous soils with ph levels near or above 7–8, although it occurs in beech woods on lime-deficient soils in the united kingdom [3]. according to czerniecki [4] some species of truffles were eaten by polish nobility in the author’s time, so it seems likely that t. aestivum sites have existed in poland for at least 300 years. some reports of attempts to cultivate truffles are present in late 19th century literature [5–7]. in the last decade, new distributional data for t. aestivum and other truffle species have been reported from poland [8,9] and neighboring countries, slovakia [10], the czech republic [11] and germany [12]. tuber macrosporum is another truffle species highly appreciated as delicacies, which has been found recently in poland [9]. this species is common in central italy while it has been reported as very rare in the czech republic, france, hungary, romania, serbia, switzerland, ukraine and the united kingdom [13]. two years ago the species was found in germany where they were considered extinct [12]. tuber macrosporum forms ectomycorrhizae with c. avellana, c. colurna and quercus species, for example: q. petraea, q. pubescens, q. robur and q. suber [15]. coniferous species, especially pinus sylvestris can also form ectomycorrhizae with t. macrosporum [15]. currently, due to growing demand for truffles, the establishment of truffle orchards in poland is in progress. some areas are especially conducive to truffle cultivation due to similarities of soil and/or climatic conditions to that of french and italian localities. truffle productivity and phenology is driven by interrelated biotic and abiotic factors, such as: climate, physiology, pedology and vegetation [14]. the data presented here have a practical application for those who are interested in truffle cultivation. the knowledge of truffles’ soil requirements and the plant species sharing the same ecological niche should be useful in choosing a suitable location for truffle orchards. material and methods the study was conducted in nida basin and przedbórz upland, the southern part of poland. the environmental characteristic of the natural forest stands is given in tab. 1. the truffle localities at the stands were found using the trained truffle dogs in collaboration with researchers from agricultural university in nitra. inventories were made in 2012–2014. yield of truffles is given in tab. 2. species of truffles were identified on the basis of microscopic features and compared to the criteria by granetti et al. [15]. samples of fruiting bodies were also taken for molecular identification. the dna sequences are deposited in genbank ncbi (accession numbers to their sequences are as follow: t. maculatum-kj524540.1, t. excavatum-kj524535.1, t. macrosporum-kj524532.1, t. aestivum-kj524527.1, t. rufum-kc330221). 269© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands over three years of study, the mean annual precipitation was 600 mm and the annual mean temperature for the same period was 8.0°c. at each of the six localities the sampling site (100 m2) was established. the vegetation at each site was surveyed in order to determine whether there was any indicator species for t. aestivum and/or co-occurring truffle species. the host-plants as well as the plants of forest floor whose biotopic preferences coincided with those of tuber spp. are summarized in tab. 3. the soil was sampled at the central part of each site by removing the litter and vegetation layers and then collecting approximately 0.5 kg of soil down to a depth of 0–30 cm, depending on the rockiness of the soil. the soil analyses were performed by the polish centre for accreditation (no. ab740). the soil ph in water (tab. 3) and essential nutrient contents were measured according to iso 10390 [17] and pb-14ed.2 of 1 january 2010 [using inductively coupled argonplasma spectrometry following mineralization in chloric (vii) acid], respectively. the percentages of n and c were analyzed according to iso13878 [17] and iso 10694 [18]. the soil texture was evaluated on the basis of occurrence of three particle size fractions: <2 μm (clay), 2–63 μm (silt), and 63–2000 μm (sand) [19]. plot no. site location bedrock altitude (m) vegetation tuber species 1 m nida basin marlstone 250–252 broadleaved forest with quercus robur, tilia cordata, carpinus betulus, fagus sylvatica and corylus avellana t. aestivum t. excavatum t. maculatum t. rufum 2 sa nida basin marlstone 311–314 oak forest with quercus petraea, carpinus betulus, cerasus avium and corylus avellana t. aestivum t. excavatum t. rufum 3 wr nida basin marlstone 290–296 broadleaved forest with quercus petraea, acer pseudoplatanus, carpinus betulus and corylus avellana t. aestivum t. excavatum 4 gr nida basin gypsium 254–261 oak forest with quercus robur and carpinus betulus t. aestivum t. excavatum 5 pr przedbórz upland jurassic limestone 312–325 beech forest with fagus sylvatica t. aestivum 6 nw nida basin marly limestone 227–228 thicket with carpinus betulus, acer campestre and populus tremula t. aestivum t. macrosporum t. maculatum t. excavatum tab. 1 environmental characteristic of tuber spp. sites. 270© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands   si te /y ea r m sa w r g r p r n w tu be r sp ec ie s 20 12 20 13 20 14 20 12 20 13 20 14 20 12 20 13 20 14 20 12 20 13 20 14 20 12 20 13 20 14 20 12 20 13 20 14 t. a es tiv um 17 74 41 17 21 11 1 15 30 3 0 0 32 7 16 6 36 8 0 14 14 7 t. e xc av at um 11 6 26 5 15 2 0 22 13 12 0 97 67 0 29 20 0 0 0 0 14 46 t. m ac ro sp or um 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 18 0 t. m ac ul at um 2 1 2 0 0 0 0 0 0 0 0 0 0 0 0 0 1 9 t. r uf um 1 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 to ta l 13 6 34 0 19 5 17 43 24 12 1 11 0 37 2 0 29 52 7 16 6 36 8 1 47 20 2 ta b. 2 y ie ld (n um be r of fr ui tb od ie s) o f t ub er s pp . w ith in th re e ye ar s at in ve st ig at ed s ite s. 271© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands results our inventory revealed five species from tuber genus: t. aestivum, t. macrosporum, t. rufum, t. excavatum and t. maculatum. the geographical names of the site where the species were found will be available for further research, but not for publication. publishing site names could lead to reckless prospecting for truffles, resulting in damage to the surrounding flora. at five out of six sites t. aestivum was accompanied by other truffle species (tab. 1, tab. 2). the stands are mixed broadleaved forest, with diverse plants of forest floor. the stand pr is rather poor in forest floor vegetation, and the only hostspecies is fagus sylvatica. only species of t. aestivum were found there. soils from the investigated localities are of rendzic type. their chemical properties are given in tab. 3. according to the atlas of forest soils of poland [21], the bedrock of the regions consists of cretaceous marlstone, limestone, gypsum and of miocene clays and sands. various quaternary deposits cover more than three quarters of the region. the analyzed soils varied from “heavy” (up to 40.23 % clay) to “light” (up to 63.41% sand) – tab. 3. the highest diversity of trees and shrubs at investigated sites were in the stand depicted as wr. this stand was the richest in regard to herbaceous plants and mosses. the lowest richness of forest floor plants and mosses was observed at the stand nw. plants associated with tuber species are given in tab. 4. number of tree and shrub host-species differ from two to six in all stands. coniferous species were represented by pinus sylvestris and its presence was noted only at stand wr. one hundred and five species of forest floor plants were identified, among them, three orchids’ species – putative host-species of truffles. they were present in all but one stand (nw). at this site the lowest number of plants of the forest floor was observed. in the stand pr no species representing mosses were noted. site measured parameter m sa wr gr pr nw mean ±sd range soil particle size fractions (%) clay 33.10 40.23 23.30 17.11 22.69 14.42 25.14 ±9.80 14.42–40.23 silt 55.83 47.03 61.70 37.32 28.33 22.17 42.06 ±15.53 22.17–61.70 sand 11.07 12.74 15.00 45.57 48.98 63.41 32.80 ±22.60 11.07–63.41 chemical characteristics ph (h2o) 7.2 7.1 7.0 7.3 7.2 7.5 7.22 ±0.17 7.00–7.5 caco3 total (%) 6.02 1.17 31.52 1.81 6.04 4.04 8.43 ±11.49 1.17–31.52 ca (%) 34.44 15.64 42.80 14.99 19.77 25.53 ±12.45 14.99–42.80 c total (%) 10.09 5.165 5.625 5.38 5.98 2.108 5.72 ±2.56 2.11–10.09 c organic (%) 9.364 5.024 4.791 5.16 5.25 1.623 5.20 ±2.46 1.62–9.36 n total (%) 0.764 0.356 0.41 0.46 0.371 0.138 0.42 ±0.20 0.14–0.76 c/n 13.2 14.5 13.7 11.7 16.12 15.30 14.09 ±1.58 11.70–16.12 tab. 3 overview of the soil properties at the tuber aestivum sites. 272© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands tree and shrubs m sa wr gr pr nw abies alba + acer campestre + acer platanoides + + acer pseudoplatanus + + + carpinus betulus + + + + + cerasus avium + + + + cornus sanguinea + + + + + corylus avellana + + + + crataegus monogyna + + + + + + daphne mezereum + euonymus europeus + + + + euonymus verrucosus + + + fagus sylvatica + + + + frangula alnus + fraxinus excelsior + + + + juniperus comunis + ligustrum vulgare + lonicera xylosteum + + + malus sylvestris + + padus avium + padus serotina + pinus sylvestris + populus tremula + prunus spinosa + + + pyrus communis + + + quercus petraea + + + + quercus robur + + + + + quercus rubra + rosa canina + + + + sambucus nigra + + + + sorbus aucuparia + + tilia cordata + ulmus glabra + ulmus minor + viburnum opulus + + + + + + host species / all species 6 / 17 4 / 18 6 / 20 5 / 16 1 / 9 3 / 14 plants of the forest floor m sa wr gr pr nw herbaceous plants actaea spicata + adoxa moschatellina + + aegopodium podagraria + + + + agrostis capillaris + + + + + + tab. 4 plant species recorded on the studied plots of tuber spp. sites known (in bold font) and potential host plants are distinguished in boxes. 273© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands ajuga reptans + + + + anemone nemorosa + + + + + asarum europaeum + + + + astragalus glycyphyllos + + + + + athyrium filix-femina + brachypodium sylvaticum + + + + calamagrostis epigejos + campanula persicifolia + campanula rapunculoides + + + campanula rotundifolia + campanula trachelium + + carex digitata + + + carex echinata + + carex hirta + carex sylvatica + + + + carex umbrosa + cephalanthera damasonium + + + + chaerophyllum aromaticum + + + chamaecytisus ratisbonensis + cimicifuga europaea + + + cirsium arvense + clinopodium vulgare + convallaria majalis + + + + cruciata glabra + + + + cypripedium calceolus + dactylis polygama + dryopteris carthusiana + dryopteris filix-mas + epipactis helleborine + fallopia dumetorum + festuca gigantea + festuca heterophylla + fragaria vesca + + galeobdolon luteum + + galeopsis pubescens + + + + galium aparine + galium boreale + + galium mollugo + galium odoratum + + + + + galium schultesii + + galium sylvaticum + + + geranium robertianium + + + geum urbanum + + + + + + tab. 4 (continued) 274© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands glechoma hederacea + hepatica nobilis + hieracium murorum + + holcus mollis + impatiens parviflora + lapsana communis + + laserpitium latifolium + lathyrus nigier + + lathyrus vernus + + + + + lilium martagon + + + luzula pilosa + lysimachia nemorum + maianthemum bifolium + + + + melica nutans + + melampyrum nemorosum + + melittis melissophyllum + + + + milium effusum + + + + moehringia trinervia + + mycelis muralis + + oxalis acetosella + paris quadrifolia + pimpinella saxifraga + plantago major + polygonatum multiflorum + + + + primula elatior + + prunella vulgaris + pulmonaria obscura + + + + + + ranunculus cassubicus + + + ranunculus lanuginosus + + rubus idaeus + rubus saxatilis + rubus nessensis + + + rubus pedemontanus + sanicula europaea + + + + + + solidago virgaurea + sonchus oleraceus + stachys sylvatica + stellaria holostea + taraxacum officinale + + + + torilis japonica + urtica dioica + veronica chamaedrys + vicia sepium + vincetoxicum hirundinaria + viola mirabilis + + + + tab. 4 (continued) 275© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands discussion truffles mainly depend on mutual relationships with angiosperm hosts including quercus spp., fagus sylvatica and corylus avellana [21,22]. however some species such as tuber puberulum berk. & broome and tuber borchii vittad. prefer gymnosperm hosts [23]. host specificity plays an important role but is not clearly defined for most wildlife associations [24]. our previous inventory suggested that, in poland, the deciduous host species are highly important to the occurrence of tuber species [9]. for example, the greatest abundance of t. aestivum fruiting bodies was found in forests where species such as: quercus robur, corylus avellana, fagus sylvatica, carpinus betulus and tilia cordata occur together [8]. our findings presented in this work changed this opinion slightly, since at pr site the only host–species tree is beech and, moreover, we obtained here the great yield of t. aestivum within the last three years. fructification of t. astivum at plots where p. sylvestris and/or p. tremula is present, are confirmation of findings by stobbe et al. [12]. the authors mentioned picea abies, abies alba, ulmus spp. and populus spp. as potential hosts. some of the above-cited studies reported the reduction of vegetation due to the presence of tuber spp. according to gryndler and co-authors [25] this could be the result of negative effect of the tuber mycelium on the growth of some of the non-host plants. there can be also direct effects (both positive and negative) of the non-host plants on the tuber mycelium. thus, the interaction between tuber mycelium and plant community could be more complex than just forming the ectomycorrhizal symbiosis. physico-chemical analysis of soils on our sites revealed that the soil texture is moderately varied, ranging from silty-clay to clayey-silt and more rarely silty, silty-sandy or clayey. the analyzed soils varied from sandy clay (up to 40.23% clay) to sandy loam up to 63.41% sand. the latter soils are generally thought to be less favorable for truffle fructification, however even excessively sandy soils can support truffle development viola reichenbachiana + + + + + + mosses atrichum undulatum + brachythecium velutinum + + eurhynchium angustiret + eurhynchium striatum + + fissidens taxifolius + hypnum cupressiforme + mnium undulatum + mnium elatum + oxyrrhynchium hians + + + pseudoscleropodium purum + rhytidiadelphus triquetrus + host species / all species 2 / 38 1 / 25 2 / 66 0 / 43 1 / 37 0 / 17 tab. 4 (continued) 276© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands if they are sufficiently rich in calcium [26,27]. our results showed that soils with the higher content of calcium, in the form of ca cations and caco3, were conducive to the higher fructification of t. aestivum. all investigated soils were poor in readily degradable nitrogen, a c/n ratio was above ten and varied from 13.2 to 16.12. this means that the development of mycorrhizae could not have been limited. development of truffles depends on ph as well. according to chevalier and frochot [28] truffles can grow in soils with ph range 6.8 to 8.0. however it cannot be claimed that ph 7.5 is the optimal level to maintain the highest level of mycorrhization on host plants roots [29]. although we did not investigate the mycorrhiza structure, our observation made on truffles’ fructification showed that t. aestivum can form fruitbodies even in soils with ph 7. the results might be really important for truffle growers, especially in countries where the area of calcareous soils is limited. the role of soil properties other than ph and carbonate content for truffle fructification is still poorly understood, so further work is needed. establishing of truffle orchards in our country has been started in a previous decade and is still in progress. we have to take into account the numerous, interrelated biotic and abiotic factors driving truffle productivity and phenology from a holistic perspective [14]. otherwise, we can derive some contradicted conclusion, for example, the greatest abundance of t. aestivum fruiting bodies was found in forests where all of host-species occurred together [8]. however, at the site indicated as pr, where the only host-species tree is beech, we found a great abundance of t. aestivum fruit bodies within the last three years (2012–2014). therefore we have to be cautious with the statement that a greater diversity of host-species occurs with a higher yield of truffles. presently, we would rather be of the opinion that each of the investigated sites is unique. as long as we do not have the climatic, soil and vegetation data combined together our conclusions will remain preliminary. acknowledgments the authors thank two anonymous reviewers for useful comments improving the manuscript. this work was due to grant financed by state forest holding no. or-2717/19/11. authors’ contributions the following declarations about authors’ contributions to the research have been made: design of the study: dh, arg; study data interpretation, manuscript preparation, literature review: dh, arg, hs. references 1. pacioni g, comandini o. tuber. in: cairney jwg, chambers sm, editors. ectomycorrhizal fungi. key genera in profile. berlin: springer; 1999. p. 163–186. http://dx.doi.org/10.1007/978-3-662-06827-4 2. chevalier g. the truffle of europe (tuber aestivum): geographic limits, ecology and possibility of cultivation. osterr z pilzkd. 2010;19:249–259. 3. pegler dn, spooner bm, young twk. british truffles. a revision of british hypogeous fungi. kew: royal botanical gardens; 1993. 4. czerniecki s. compedium ferculorum albo zebranie potraw. polish first cookbook. kraków: collegium columbinum, szedlowie jerzy i mikołaj; 1682. 5. aleksandrowicz j, błoński f. encyklopedia rolnicza. warszawa; 1894. 6. gawarecki z. trufle i ich sztuczne pielęgnowanie. lwów: red. “bartnika postępowego”; 1895. 7. spausta w. trufle. sylwan. 1897;15(6,7):161–167,201–208. http://dx.doi.org/10.1007/978-3-662-06827-4 277© the author(s) 2014 published by polish botanical society acta mycol 49(2):267–277 hilszczańska et al. / truffles in natural stands 8. hilszczańska d, sierota z, palenzona m. new tuber species found in poland. mycorrhiza. 2008;18(4):223– 226. http://dx.doi.org/10.1007/s00572-008-0175-4 9. hilszczańska, d, rosa-gruszecka a, sikora k, szmidla h. first report of tuber macrosporum occurrence in poland. scientific research and essays. 2013;7(23):1096–1099. http://dx.doi.org/10.5897/sre2013.5529 10. gazo j, miko m, chevalier g. first results of inventory research on economically important species of truffles (tuber) in the tribec mountains. acta fytotechnica et zootechnica. 2005;8(3):66–71. 11. streiblova e, gryndlerova h, valda s, gryndler m. tuber aestivum – hypogeous fungus neglected in the czech republic. a review. czech mycol. 2010;61(2):163–173. 12. stobbe u, büntgen u, sproll l, tegel w, egli s, fink s. spatial distribution and ecological variation of re-discovered german truffle habitats. fungal ecol. 2012;5(5):591–99. http://dx.doi.org/10.1016/j. funeco.2012.02.001 13. hall i, brown g, zambonelli a. taming the truffle. the history, lore, and science of the ultimate mushroom. portland, or: timber press; 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(soil biology, vol 34). http://dx.doi.org/10.1007/978-3-642-33823-6_10 28. chevalier g, frochot h. la truffe de bourgogne. paris: pétrarque; 1997. 29. thomas pw. the role of ph in tuber aestivum syn. uncinatum mycorrhiza development within commercial orchards. acta mycol. 2012;47(2):161–167. http://dx.doi.org/10.5586/am.2012.019 http://dx.doi.org/10.1007/s00572-008-0175-4 http://dx.doi.org/10.5897/sre2013.5529 http://dx.doi.org/10.1016/j.funeco.2012.02.001 http://dx.doi.org/10.1016/j.funeco.2012.02.001 http://dx.doi.org/10.1038/nclimate1733 http://dx.doi.org/10.1016/0167-8809(90)90016-7 http://dx.doi.org/10.1111/j.1365-294x.2010.04855.x http://dx.doi.org/10.1111/j.1365-294x.2010.04855.x http://dx.doi.org/10.1093/icb/42.2.352 http://dx.doi.org/10.1007/s00572-014-0580-9 http://dx.doi.org/10.5586/am.2012.014 http://dx.doi.org/10.1007/978-3-642-33823-6_10 http://dx.doi.org/10.5586/am.2012.019 abstract introduction material and methods results discussion acknowledgments authors’ contributions references 2014-12-31t17:41:55+0000 polish botanical society 2014-11-20t23:14:21+0000 polish botanical society the genus echinostelium (myxomycetes) in lithuania gražina adamonytė laboratory of mycology, institute of botany žaliųjų ežerų 49, lt 08406 vilnius, grazina@botanika.lt a d a m o n y t ė g.: the genus echinostelium (myxomycetes) in lithuania. acta mycol. 41 (2): 169 176, 2006. seven species of the genus echinostelium apitectum, e. arboreum, e. brooksii, e. colliculosum, e. corynophorum, e. aff. elachiston, e. minutum are reported from lithuania. their morphological peculiarities are discussed; a key to the species, pictures and distribution maps are given. key words: myxomycetes, acellular slime molds, moist chamber cultures, morphology, ecology, distribution introduction the genus echinostelium de bary was erected in r o s t a f i ń s k i (1873) for myxomycetes with very pale spores and stipitate minute sporocarps that rarely exceed 500 μm. myxomycetes of the genus mostly inhabit bark of living trees and shrubs but can also be occasionaly found on litter including tiny branchlets or coniferous needles. their sporocarps are especially fragile and usually short-living, therefore they are seldom collected in the field. thus, for investigations of this group simple but highly efficient techniques of substrate incubation in the laboratory is very useful. so far 16 species have been described within the genus. one of them, e. vanderpoellii nann.-bremek., d. w. mitch., t. n. lakh. et r. k. chopra, later was proposed to reduce to a synonym of e. apitectum k. d. whitney (p a n d o 1997). another one, echinostelium roseum ing, was excluded because it appeared to be not a myxomycete (i n g 1984). in lithuania the first records of echinostelium species were obtained in late 1990‘ies; up to now 7 species are known from the country: echinostelium apitectum, e. arboreum, e. brooksii, e. colliculosum, e. corynophorum, e. aff. elachiston, e. minutum. all these species except for e. minutum are reported here for the first time for lithuania. the paper also presents morphological, ecological and geographical data on lithuanian echinostelium species. acta mycologica vol. 41 (2): 169-176 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 170 g. adamonytė material and methods virtually all echinostelium specimens described here were obtained from moist chamber cultures. w h i t n e y (1980) proposed a special protocol for revealing echinostelium species; it includes substrate soaking for 1–3 hours, and further incubation in the dark at 12–15°c. in the present research the cultures were processed following h ä r k ö n e n (1977) because i aimed to reveal not only echinostelium, but all myxomycetes which might inhabit a particular substrate. so, bark pieces cut from a living tree/shrub trunk or main branches were placed in one layer into petri dishes lined with filter paper. the dishes were filled with distilled water and left closed for 24 hrs at room temperature in a natural light regime, then excess water was poured out. the dishes closed with covers were further kept in room temperature in a natural light regime and regularly checked for myxomycete sporocarps – on the first incubation week daily, later on once a week. emerged sporocarps were allowed to dry slowly by slightly opening a lid and leaving for a night. echinostelium species usually developed within first few days, but sometimes additional mass sporifications of e. minutum were observed after a considerable time. microscopic examination was carried out in fresh preparations in 3% koh. micrographs of sporocarps stained with cotton blue were made with a pentax *istds camera mounted on a biolam–i microscope. scanning electron micrographs were made from air-fresh material with hitachi s2500 sem at the natural history museum, london. voucher specimens of the species are kept in the herbarium of the institute of botany, vilnius (bilas). bark ph was measured with iq–150 ph-meter with thermorussel flat-head electrode kdcef11 on the second day after water was removed. nomenclature of myxomycetes follows l a d o (2001). standard forms of authors’ names are according to b r u m m i t t and p o w e l l (1992). species descriptions and discussion echinostelium apitectum k. d. whitney, mycologia 72 (5): 954 (1980), fig. 1, 2, 3, 4. sporocarps gregarious, rosy when fresh, later turning whitish, 120-300 μm high; stalk hyaline in transmitted light (tl), partly filled with a refuse material; sporotheca 40-70 μm diam., spores closely packed together; peridium persisting as a basal collar covering up to 1/3 of a spore-like body; spore–like body 8-14 μm diam.; columella mostly reduced or inconspicuous; capillitium absent, when present reduced to a short single or forking thread; spores whitish in mass, hyaline in tl, smooth or minutely warted, 6-9.5 μm diam. substrates. bark of fraxinus excelsior, picea abies, pinus sylvestris, populus sp., quercus robur, ulmus sp. substrate ph ranges from 3.4 to 6.9. distribution. kėdainiai, pasvalys, ukmergė, jonava, prienai distr., vilnius city (fig. 5). frequent, more than 80 records. notes. echinostelium apitectum is rather variable species ranging from well-developed columella (bearing threads of capillitium) to strongly reduced or even absent columella. l a d o and p a n d o (1997) distinguish two forms of e. apitectum: one with large (10-12 μm diam.) and the second with small (6-9 μm diam.) spores, sporocarps of the latter also being taller and more slender. but the authors admit that the genus echinostelium (myxomycetes) in lithuania 171 both forms merge, and for their taxonomical recognition further evidence would be needed. in the lithuanian material of e. apitectum two groups can be distinguished, too. one group included stouter sporocarps with no apparent columella, spore-like body reaching 11-14 μm diam., and spores approx. 6-7.5 μm diam., appearing warted under transmitted light (tl, oil immersion). the other one covered higher and more slender sporocarps with a smaller spore-like body (8-10 μm diam.), discernible columella, and slightly larger spores (7.5-8 μm diam.). but, similarly to l a d o and p a n d o (1997) experience, there were also specimens transitional between both groups in the lithuanian material, therefore all they were ascribed to e. apitectum. in lithuania e. apitectum was most frequently found on acid substrates: the highest number of collections was obtained from pinus sylvestris bark which ph ranged from 3.7 to 4.6. the myxomycete was also found – albeit only sporadically – on bark of deciduous trees with higher ph (up to 6.3); one collection was found on bark with nearly neutral ph reaching 6.9. this experience rather supports results obtained by wr i g l e y d e b a s a n t a (2004): in her model experiments of acid rain simulation e. apitectum sporulated on bark with lower ph values after treating it with solutions of ph 3 and 4. however, some authors report that e. apitectum was frequently collected from bark of juniperus thurifera (l a d o 1993) and olea europaea (p a n d o 1989, l. c. wr i g l e y d e b a s a n t a 2000) whose ph is significantly higher – 5.5-6.5. echinostelium arboreum h. w. keller et t. e. brooks, mycologia 68: 1207 (1977), fig. 6. sporocarps scattered, sitting on leaf tips of mosses, yellow, 130-150 μm high; stalk yellowish in tl, partly filled with a refuse material; sporotheca 70-80 μm diam.; peridium persistent, shining, when evanescent remains as a colar at the base of columella; capillitium well developed, branching dichotomously up to 3 times, not forming a periferial net; spores hyaline in tl, varted, 8.5-9 μm diam. substrates. bark of fraxinus excelsior overgrown with epiphytic mosses neckera complanata (biržai distr.) and leucodon sciuroides (ukmergė distr.). ph 5.8. distribution. biržai, ukmergė distr. (fig. 7). rare, 3 records. notes. species is easily recognizable by bright-yellow short-stalked sporocarps with shining peridium and abundant capillitium. in both localities, bark for moist chamber cultures was collected in biologically rich forests. in biržai district it was collected in the botanical reserve of biržai forfig. 5. localities of e. apitectum (■), e. colliculosum (●), and e. aff. elachiston (♦) in lithuania. 172 g. adamonytė est, in ukmergė district bark was taken in a broad-leaved forest with quercus robur ca 150 years-old. echinostelium brooksii k. d. whitney, mycologia 72 (5): 957 (1980), fig. 8. sporocarps gregarious, rosy when fresh, turning pale brown, 100-150 μm high; stalk hyaline in tl, partly filled with a refuse material; sporotheca 40-60 μm diam., spores loosely packed in the sporotheca; peridium evanescent, remaining as a small colar at the base of columella; columella hemispherical on a short stalk, brown, 4.56.5 μm diam.; spores rosy in mass, pale rosy in tl, appearing smooth, with a thinner germination area, 10.5-14 μm diam. substrates. bark of picea abies, pinus sylvestris, occasionally fraxinus excelsior. substrate ph ranges from 3.4 to 5.7. distribution. jonava, kėdainiai, prienai, trakai distr. (fig. 7). frequent, more than 75 records. notes. echinostelium brooksii is close to e. corynophorum; for differences columella and spores should be examined (see comments under e. corynophorum). e. brooksii most frequently occurred on bark of pinus sylvestris, together with echinostelium apitectum and e. minutum. e. brooksii sporulated with the highest frequency on bark whose ph range was the same as for e. apitectum – from 3.7 to 4.6, but its general ph range was narrower: only a few collections were obtained from bark which ph was more than 5.0. so, this species appears to be confined to the most acid substrates among species of the g. echinostelium. echinostelium colliculosum k. d. whitney et h. w. keller, mycologia 72: 641 (1980), fig. 9, 10, 11. sporocarps gregarious, whitish, 70-120 μm high; stalk hyaline in tl, partly filled with a refuse material; sporotheca 30-40 μm diam.; peridium persisting as a colar; spore-like body with thickened areas, 8.5-9 μm diam.; spores hyaline in tl, minutely warted, bearing circular thickened areas, 8-9.5 μm diam. substrate. bark of fraxinus excelsior. ph 6.6–7.5. distribution. akmenė distr., vilnius city (fig. 5). rare, 4 records. notes. echinostelium colliculosum is characterized by small sporocarps and thickened articular areas on the spore wall. from a very closely related species e. coelocephalum t. e. brooks et h. w. keller (which have not been registered in lithuania, so far) it is said to differ in larger spores with less pronounced thickened areas, as fig. 7. localities of e. arboreum (■), e. brooksii (●), and e. corynophorum (♦) in lithuania. the genus echinostelium (myxomycetes) in lithuania 173 well as in the colar form (w h i t n e y , k e l l e r 1980). thus, in e. colliculosum collar is larger and its margins adhere to the spore-like body, while in e. coelocephalum collar margins appear to stay free. in specimens which are described here the colar was large, and its margins were attached closely to the spore-like body. but even under oil-immersion it was difficult to discern whether thickened areas on a spore-like body and spore walls were of the uniform thickeness (e. coelocephalum) or tapering towards edges (e. colliculosum). as the critical drying point technique was not applied while preparing material for sem examination, these thickenings were not distinct in sem photographs, too. in lithuania e. colliculosum was observed on bark of trees growing along roadsides; ph of the bark cultures was close to neutral. bearing in mind that in western kazakhstan steppe e. colliculosum was also collected from windbreak-forming trees with bark ph as high as 7.2–8.4 (unpublished data), it appears that this species prefers substrata with neutral to slightly alkaline reaction. echinostelium corynophorum k. d. whitney, mycologia 72: 963 (1980), fig. 12. sporocarps gregarious, white, up to 100 μm high; stalk hyaline in tl, partly filled with a refuse material; sporotheca ca. 30 μm diam.; peridium remaining as a small colar at the base of columella; spore-like body absent; columella subglobose, on a short stalk, light brown, 3-3.5 μm diam., 3.5-4 μm high; spores hyaline in tl, with thickened areas, 11.5-12 μm diam. substrate. alnus glutinosa female cones; ph 6.1. distribution. tauragė distr. (fig. 7). rare, 1 record. notes. as noted by w h i t n e y (1980) echinostelium corynophorum is closely related to e. brooksii. the author points at the following differences: columella in e. corynophorum is hyaline to pale yellow while in e. brooksii it is always deeply dark; spores of e. corynophorum bear thickenings and are white, meanwhile spores of e. brooksii are smooth and rosy. for distinguishing these two species l a d o and p a n d o (1997) suggest one more particular trait: the thinnest part of e. corynophorum stalk is in a short distance below the collar, and the thinnest section of e. brooksii stalk is right below the colar. in the only specimen from lithuania which is described here the thinnest area of the stalk was not well distinguished, the size of sporotheca and columella were on the smaller end of the scale for the species, but spores bore distinct thickened areas. the shape of columellae of e. brooksii and e. corynophorum collected in lithuania differed markedly: the first was hemispherical, or horizontally lenticular, and the second was subglobose. echinostelium aff. elachiston alexop., mycologia 50: 52 (1958), fig. 13. sporocarps gregarious, whitish, shining, 100-110 μm high; stalk yellowish in tl, partly filled with a refuse material; sporotheca 30-35 μm diam; peridium hyaline, after evanescing leaving a large collar (ca 15 μm) on the top of stalk; spore–like body absent; columella indiscernible; spores appearing warted (oil-immersion), 8-9.5 μm diam. substrate. bark of fraxinus excelsior. distribution. biržai distr. (fig. 5). rare, 2 records. notes. echinostelium elachiston is characterized by small, yellow tinted sporocarps, a wide collar on the tip of stalk, scanty to absent capillitium, and spores of 6.5-8 μm diam. m a r t i n and a l e x o p o u l o s (1969) state that spores of this species 174 g. adamonytė are smooth with well-marked thickened circular areas on the wall, while w h i t n e y (1980) specifies that they are minutely roughened and lacking circular thickenings. spores of specimens from spain described by l a d o and p a n d o (1997) also are said to have smooth wall of uniform thickness, but their measurements reach up to 11 μm diam. warts on spore wall of both available lithuanian specimens were very conspicuous, particularly when stained with cotton blue, and spores were in general larger than it is noted in the species protologue. all other characteristics of these specimens rather well agreed with the concept of e. elachiston. substrate ph was not measured for available specimens of e. aff. elachiston, but data show that ph of bark of fraxinus excelsior growing in natural conditions is close to 5.5-6 (unpublished data). echinostelium minutum de bary in rostaf., śluzowce monogr.: 215 (1874), fig. 14. sporocarps gregarious, white or pale rosy, 250-500 μm high; stalk hyaline in tl, partly filled with a refuse material; sporotheca 50 μm diam.; peridium evanescent, remaining as a small colar at the base of columella; spore-like body absent; columella light brown, ca 4 μm high; capillitium well developed, never forming a net, consisting of a few threads, usually one or two of them being long and dichotomously branched; spores hyaline or pale rosy, 6.5-14 μm diam. substrates. bark of alnus glutinosa, betula sp., fraxinus excelsior, juniperus communis, picea abies, pinus sylvestris, populus tremula, quercus robur; occasionally litter: female cones of alnus glutinosa, mixed litter of leaves, fine branchlets and needles; once excrements of herbivores (moose). substrate ph ranges from 3.4 to 6.9. distribution. biržai, jonava, kėdainiai, lazdijai, prienai, radviliškis, šalčininkai, tauragė, trakai, ukmergė, varėna distr., neringa city (fig. 15). common, more than 120 records. notes. small sporocarps of echinostelium minutum with scanty capillitium can resemble e. apitectum, however the latter species has a spore-like body. e. minutum is the most common species of the genus recorded in almost all regions of lithuania where myxomycetes were investigated. its sporocarps readily appeared in moist chamber cultures on a great variety of substrata with a wide range of ph from highly acidic to nearly neutral. but most frequently its sporification was observed at ph 3.4–6.1. if the cultures were kept for sufficiently long time, additional waves of e. minutum sporification occured. e. g, in a culture of moose dung sporocarps of this species were noted 3 months after setting the culture, then the fig. 15. localities of echinostelium minu tum (■) in lithuania. the genus echinostelium (myxomycetes) in lithuania 175 next sporification occurred three and a half months after the first sporification. one more sporification took place 10 months after setting the culture, but sporocarps were scanty. this phenomenon was not observed for other echinostelium species, although it was noted for physarum viride (bull.) pers. var. aurantium (bull.) lister, arcyria cinerea (bull.) pers., and paradiacheopsis fimbriata (g. lister et cran) hertel (d v o ř á k o v á 2002). key to the genus echinostelium lithuania 1. capillitium present ...................................................................................................... 2 – capillitium absent . ....................................................................................................... 4 2. capillitium well developed, spore-like body absent ................................................... 3 – capillitium scanty, spore-like body present ............................................... e. apitectum 3. sporocarps long-stalked, white or rosy ....................................................... e. minutum 3. sporocarps short-stalked, yellow .............................................................. e. arboreum 4. spore-like body present .............................................................................................. 5 – spore-like body absent ................................................................................................. 6 5. spore wall with circular thickenings .................................................... e. colliculosum – spore wall without circular thickenings .................................................... e. apitectum 6. columella present ....................................................................................................... 7 – columella absent ....................................................................................... e. elachiston 7. columella dark, spore wall without circular thickenings ............................ e. brooksii – columela pale, spore wall with circular thickenings ......................... e. corynophorum acknowledgements: this paper is dedicated to an outstanding polish mycologist professor alina skir giełło. i much appreciate help of dr. carlos lado at determination of a specimen of echinostelium apitec tum. thanks are due to dr. ernestas kutorga for discussions during preparation of the paper, and to dr. ilona jukonienė for identification of mosses. this study was supported in part by the lithuanian studies and science foundation (grants no g 139, g 169, t 79/05). access to sem and photomicrography and light microscopy facilities at the natural history museum, london, was granted by the ec funded ihp programme sys resource. many thanks are extended to mr. chris jones for introducing me sem techniques. references b r u m m i t t r . k . , p o w e l l c . e . 1992. authors of plant names. kew. d v o ř á k o v á r. 2002. myxomycetes in bohemian karst and hřebeny mts. czech mycol. 53 (4): 319 349. h ä r k ö n e n m . 1977. corticolous myxomycetes in three different habitats in southern finland. karstenia 17: 19 32. i n g b . 1984. on the identity of echinostelium roseum b. ing (myxomycetes). trans. br. mycol. soc. 82: 173. l a d o c . 1993. myxomycetes of mediterranean woodlands. (in:) d . n . p e g l e r , l . b o d d y , b . i n g , p. m . k i r k (eds). fungi of europe: investigation, recording and conservation. royal bo tanical gardens, kew: 93 114. l a d o c . 2001. nomenmyx. a nomenclatural taxabase of myxomycetes. real jardín botánico (csic), madrid. l a d o c . , p a n d o f. 1997. myxomycetes, i. ceratiomyxales, echinosteliales, liceales, trichiales. real jardín botánico (csic) & j. cramer, madrid, berlin, stuttgart. m a r t i n g . w. , a l a x o p o u l o s c . j . 1969. the myxomycetes. university of iowa press, iowa city. 176 g. adamonytė p a n d o f. 1997. a new species and a synonymy in echinostelium (myxomycetes). mycotaxon 64: 343 348. r o s t a fi ń s k i j . t. 1873. versuch eines systems der mycetozoen. strassburg. w h i t n e y k . d . 1980. the myxomycete genus echinostelium. mycologia 72: 950 987. w h i t n e y k . d . , k e l l e r h . w. 1980. a new species of echinostelium. mycologia 72: 640 643. w r i g l e y d e b a s a n t a d . 2000. acid deposition in madrid and corticolous myxomycetes. stapfia 73, zugleich kataloge des oö. landesmuseums neue folge 155: 113 120. w r i g l e y d e b a s a n t a d . 2004. the effect of simulated acid rain on corticolous myxomycetes. syst. geogr. pl. 74: 175 181. rodzaj echinostelium (myxomycetes) na litwie s t r e s z c z e n i e na litwie stwierdzono dotychczas występowanie siedmiu gatunków z rodzaju echinoste lium: e. apitectum, e. arboreum, e. brooksii, e. colliculosum, e. corynophorum, e. aff. elachi ston i e. minutum. praca zawiera klucz do oznaczania gatunków, krytyczną analizę cech mor fologicznych oraz dane o substracie i rozmieszczeniu poszczególnych gatunków z wykazaniem stanowisk na mapie litwy. 2014-01-01t11:44:07+0100 polish botanical society armillaria ectypa, a vulnerable indicator of mires esteri ohenoja botanical museum, university of oulu p.o. box 3000, fi 90014, esteri.ohenoja@oulu.fi o h e n o j a e.: armillaria ectypa, a vulnerable indicator of mires. acta mycol. 41 (2): 223 228, 2006. a boreal montane basidiomycete, armillaria ectypa, occurs as occasional in northern finland, mainly in the aapa mire area. according to the iucn criteria it has been classified as a vulnerable fungus in finland. its ecology is in some way connected e.g. to the carices of wet mesotrophic mires. key words: armillaria ectypa, agaricales, marasmiaceae, tricholomataceae, ecology, threat introduction the genus armillaria belongs to the basidiomycete family marasmiaceae (tricholomataceae), agaricales. there occur all four species of the genus armillaria in finland (a. borealis, a. cepistipes, a. ectypa, a. ostoyae). a. borealis is common in finland, rarer in the northernmost lapland, a. cepistipes is occasional, and a. ostoyae is rare occurring in southern and central finland. a. ectypa has, on the contrary, a northern distribution (o h e n o j a 1996). description of the species armillaria ectypa has (fig. 1) 5-10 cm high fruit bodies, cap 2-5 cm in diameter, hygrophan, somewhat glossy or fatty, pale ochraceous, brownish when dry, centre covered by faint dark scales or striae, margin later translucently striated. gills decurrent, rather narrow, whitish first, later ochraceous. stem slender, 5-10 x 0,5-1,5 cm, lower part often thicker (depending on the consistence of substrate), fibrillose, of the same colour as the cap, base felty. flesh watery, whitish, taste mild, smell fungous or slightly anisate. basidia 4-spored, 32-40 x 6-9 μm in size, sterigmata 2-4 μm long. spore print white or creamish, spores subglobose or ovoid, 7.0-9.0 x 5.5-6.5 μm. clamps not seen. acta mycologica vol. 41 (2): 223-228 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 224 e. ohenoja ecology and distribution armillaria ectypa has been found in finland mainly in the northern parts of the country (fig. 2), its occurrence concentrating to the aapa mire zone. typical habitats are mesotrophic, thin-turfy, often rocky or stony pine bogs or fens and alluviated shores of ponds (fig. 3). it often grows on wet substrate, on tufts of decaying carices, as carex rostrata, c. lasiocarpa and c. aquatilis. the vascular plants, mosses and fungi found in the closest neighbourhood of a. ectypa are listed in tab. 1. the southernmost known localities of armillaria ectypa are in the southern coast of finland, but those two habitats at vantaa (tab. 2) have not been checked since the species was found in 1959 and 1966. from janakkala it has been collected last time in the year 1982. this mire is still in good condition and protected, but the occurrence of a. ectypa has not been annually monitored. some habitats in the northern mires are spoiled in some amount because of draining bogs or constructing reservoirs, but very few of them have been studied later. at least one of them is totally flooded into a reservoir. about one half of the localities in finland are situated in nature reserves. 357,7535 s muonio rahtusenjärvi 360,7457 a kolari sieppijänkkä 377,7171 a raahe rytilampi 381,6685 s vantaa kaivoksela 381,6766 a janakkala suurisuo 383,6686 s vantaa sillböle 388,7447 s kolari pasmajärvi 389,7440 s kolari kaakkuririipilampi 390,7350 a tornio sorvasvaara 409,7348 s tervola ruuttulampi 410,7450 s rovaniemi lohiniva 412,7418 s rovaniemi marrasjärvi 414,7281 s kuivaniemi ihanalampi 416,7071 s haapajärvi kurikkalampi 425,7240 a haukipudas syvä susijärvi 435,7262 a ii rytisuo 452,7235 s haukipudas kallioselkä 453,7222 a kiiminki kourilampi 493,7353 a ranua tyyräsenjärvi 499,7228 s pudasjärvi koirapuro 500,7388 a kemijärvi sammalvaara 503,7734 s utsjoki kenesjärvi 508,7451 a pelkosenniemi ahma aapa 510,7451 g pelkosenniemi korpela 524,7544 a sodankylä posoaapa 538,7200 a puolanka vantaslahti 539,7265 s taivalkoski tutulampi 539,7446 a savukoski kätkäaapa 543,7231 s pudasjärvi huosiosoja 569,7349 a posio riisitunturi 576,7461 a savukoski sattoaapa 583,7418 a salla aatsinginhauta 603,7367 a kuusamo rytilampi 604,7364 a kuusamo hiidenlammet 607,7239 a suomussalmi takkosenlampi 609,7340 a kuusamo vaimojärvi 715,6949 a ilomantsi kuikkalampi fig. 2. distribution of armillaria ectypa in finland. (grid 27°e) found after the year 1980 (a) found before the year 1980 (s) extinct (g) armillaria ectypa 225 ta b l e 1 vascular plants, mosses and fungi found in the closest neighbourhood of armillaria ectypa in finland vascular plants carex aquatilis carex chordorrhiza carex echinata carex lasiocarpa carex limosa carex magellanica carex panicea carex rostrata carex rotundata phragmites australis molinia caerulea eriophorum gracile eriophorum latifolium trichophrum alpinum trichophorum cespitosum dactylorhiza incarnata salix lapponum betula nana betula pubescens ranunculus hyperboreus menyanthes trifoliata drosera anglica andromeda polifolia vaccinium oxycoccus pedicularis palustris equisetum fluviatile selaginella selaginoides mosses sphagnum angustifolium sphagnum lindbergii sphagnum majus sphagnum platyphyllum sphagnum recurvatum sphagnum subsecundum aulacomnium palustre cinclidium subrotundum loeskypnum badium paludella squarrosa pseudocalliergon trifarium scorpidium revolvens scorpidium scorpioides straminergon stramineum warnstorfia exannulata warnstorfia procera warnstorfia sarmentosa fungi laccaria proxima lactarius pubescens lyophyllum palustre ta b l e 2 the localities of armillaria ectypa in finland abrreviations: u inl biological provinces; h herbarium of the university of helsinki, joe her barium of the university of joensuu, oulu herbarium of the university of oulu, tur a herbarium of the åbo akademi . uusimaa (u). vantaa 30.8.1959 (h), 18.9.1966 (h) etelä-häme (eh). janakkala 16.9.1957 (h), 4.9.1971 (h), 2.9.1982 (h), 7.9.1982 (h) pohjois-karjala (pk). ilomantsi 27.8.1996 (h, joe) keski-pohjanmaa (kp). haapajärvi 22.7.1970 (oulu), raahe 15.9.1983 (oulu). kainuu (kn). puolanka 2.8.1989 (oulu), 25.7.1990 (oulu) oulun pohjanmaa (op). haukipudas 3.8.1968 (oulu), 17.8.1988 (oulu), kuivaniemi 29.7.1970 (oulu), pudasjärvi 24.7.1975 two finds (oulu), 9.8.1977 (oulu), ii 6.8.1985 (oulu), kiiminki 1.9.1999 (oulu). perä-pohjanmaa (pep). rovaniemi rural commune 23.7.1976 (oulu), 24.7.1976 (oulu), tervola 29.7.1978 (oulu), ranua 30.7.1992 (oulu), kemijärvi 26.7.1993 two finds (oulu), tornio 20.9.1998 (oulu). koillismaa (ks). taivalkoski 6.8.1979 (oulu), kuusamo 25.7.1986 (oulu), 20.8.2005 (oulu), 21.8.2005 (oulu), 24.8.2005 (tur-a), 1.9.2005 (oulu), posio 23.8.2005 (oulu), salla 20.7.1994 (oulu). kittilän lappi (kil). muonio 7.8.1970 (arch. oulu), kolari 31.7.1970 two finds (oulu), 29.8.1997 (arch. oulu) sompion lappi (sol). pelkosenniemi 3.8.1984 two finds (oulu), 23.7.1995 (oulu), savukoski 23.7.1995 two finds (oulu), 10.8.2001 (oulu). inarin lappi (inl). utsjoki 14.9.1972 (oulu). 226 e. ohenoja the 44 samples from finland are from 30 years between 1959 and 2005, mostly one find per season, but there are 3-5 finds from the years 1970, 1995 and 2005. a special study has not been done and the fungus has been found usually in connection of the flora and vegetation inventories. as for the phenology of the species its finds have been made in finland in july – september (20th july – 18th september), the main fruiting time being in the southern half of the country somewhat later than in the north. armillaria ectypa is, according to the criteria of iucn, a vulnerable fungus in finland (r a s s i et al. 2001) and a good indicator of the mesotrophic mire habitats. armillaria ectypa was found in 12 european countries (d a h l b e r g , c r o n e b o r g 2003), but it is rare everywhere, also in finland where it has, however, more localities than elsewhere in europe. in sweden the threat class is nt, near threatened, in denmark en, endangered. it has never been seen in norway neither in estonia nor iceland. it is considered extinct in switzerland and poland, and in mecklenburgvorpommern it had been seen last time in the year 1959 (k r e i s e l 1992). it is one of the 33 fungus species proposed to the bern convention (d a h l b e r g , c r o n e b o r g 2003). discussion distribution of armillaria ectypa seems to be boreal-montane and maybe also continental. the ecology of it is different from the other species of the genus, and very poorly known, thus far. it´s mycelium grows in very wet substrate and can survive in low oxygen conditions. some species of armillaria can oxygenate its rhizomorphs in very wet conditions (a i n s w o r t h 2003), but no rhizomorphs have been detected in a. ectypa. there grows in canada a. sinapina except on wood also in wet fens (t h o r m a n n et al. 2001), where its mycelium has been isolated from the rhizomes and decomposing leaves of carex aquatilis, and also from decomposing salix planifolia leaves. some species of armillaria can form a mutualistic relationship with the rhizomes of e.g. orchids. t h o r m a n n et al. (2001) think that a possible relationship can also exist between a. ectypa and carex aquatilis. if this kind of mycorrhizal connection really exists, it is very likely that also c. rostrata and some other carices could be suitable symbionts. there are in the bases of stems in our samples often roots, stems and leaves of carices and salices, but no proper study has been done in the field. such study is needed, for which the habitats in northern finland should suit well. except for the ecology, the homothallism of the species also is its special characteristic deviating from the other species of the genus. c h i l l a l i et al. (1998) suppose that armillaria ectypa separated, as a. tabescens, too, earlier in the evolution than the other species of the genus. a question arises if this homothallic mode of reproduction is limiting its ecological capacity so that its impact to changing environment were weaker than that of the heterothallic species? on the contrary the genesis of new mycelia could in theory be easier for these kinds of fungi. armillaria ectypa 227 references a i n s w o r t h a.m. 2003. report on the marsh honey fungus, armillaria ectypa, a uk bap species. english nature 540: 1 23. c h i l l a l i m., w i p f d., g u i l l a u m i n j. j., m o h a m m e d c., b o t t o n b. 1998. delineation of the european armillaria species based on the sequences of the internal transcribed spacer (its) of ribosomal dna. the new phytologist 138: 553 561. c r o n e b o r g h., d a h l b e r g a., h a l l i n g b ä c k t. 2002. sällsynta svampari i sverige. arter som kan komma att bli rödlistade år 2005. artdatabanken, slu. 72 pp. d a h l b e r g a., c r o n e b o r g h. 2003. 33 threatened fungi in europe. complementary and revised information on candidates for listing in appendix i of the bern convention. 82 pp. k r e i s e l h. 1992. rote liste der gefährdeten grosspilze mecklenburg vorpommerns. die umweltmi nisterin des landes mecklenburg vorpommern. 46 pp. o h e n o j a e. 1996. nevamesisieni, armillaria ectypa, suomessa. (summary: armillaria ectypa in fin land.). sienilehti 48 (3): 88 90. r a s s i p., a l a n e n a., k a n e r v a t., m a n n e r k o s k i i. (eds). 2001. suomen lajien uhanalaisuus 2000. suomen ympäristö 495: 1 432. helsinki. t h o r m a n n m.n., m y r h o l m c.l., m a l l e t t k.i. 2001. armillaria sinapina in herbaceous plant material from a peatland in alberta, canada. can. j. bot. 79 (5): 643 647. armillaria ectypa, zagrożony gatunek wskaźnikowy torfowisk s t r e s z c z e n i e armillaria ectypa, gatunek borealno górski, znany jest z licznych stanowisk w północnej finlandii. zajmuje wilgotne siedliska, często spotykany jest w darniach, na obumarłych pę dach turzyc, głównie carex rostrata i c. aquatilis. w pracy zestawione są rośliny naczyniowe, mszaki i grzyby towarzyszące a. ectypa, jak również rozmieszczenie stanowisk tego gatunku na terenie finlandii. 2014-01-01t11:44:21+0100 polish botanical society error 404 error 404 you have been redirected to this page because something went wrong... it is possible that the page you are looking for is temporarily unavailable. you may want to visit again: polish botanical society polish botanical society journals or report a problem to the webmaster at admin@pbsociety.org.pl photo: hisks   micromycetes on climbing roses leaves (rosa l.) in the botanic garden of the jagiellonian university in cracow 1 of 9published by polish botanical society acta mycologica original research paper micromycetes on climbing roses leaves (rosa l.) in the botanic garden of the jagiellonian university in cracow maria kowalik*, klaudia duda-franiak department of plant protection, faculty of biotechnology and horticulture, university of agriculture in krakow, al. 29 listopada 54, 31-425 cracow, poland * corresponding author. email: m.kowalik@ogr.ur.krakow.pl abstract micromycetes inhabiting the leaves of 20 cultivars of climbing roses (rosa l.), grown in botanic garden of the jagiellonian university in cracow was investigated in the three successive years of research. sixty-five taxa of of micromycetes was recorded with a few species dominating: alternaria alternata, epicoccum nigrum, pestalotia rosae, penicillium brevicompactum and sordaria fimicola, accompanied by various other microfungi. a high abundance of rose black spot caused by diplocarpon rosae was also observed. the affected leaves revealed advancing necrosis, substantially enhancing at the end of the growing season. defoliation took place from june to october. micromycetes inhabiting the leaves of climbing roses in botanic garden of the jagiellonian university in cracow considerably deteriorated the decorative aspect of the plants. keywords microfungi; necrosis of leaves; rose black spot; defoliation this issue of acta mycologica is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. introduction botanic gardens form a special place within a city, established to conserve plants around the world and to support teaching, research and recreation. the aesthetic experience offered by the plant collections in botanic gardens contribute to raising public awareness about the threat posed by microorganisms that cause various diseases and thus reduce the decorative aspects of plants. climbing roses make an elegant, beautiful covering for nearly any horizontal or vertical structure. the climbing roses found in the botanic garden of the jagiellonian university in cracow, represent a collection of old and rare cultivars, most of which, are no longer in cultivation. collection found in green spaces, with their low biological stability and continuous and intensive pest pressure, are highly susceptible to infections caused by mycobiota, among which micromycetes play an important role [1–3]. micromycetes inhabiting climbing roses cause deformities of above-ground parts of plants, dieback of shoots and flowers and dieback and premature leaf fall. climbing roses deprived of lush foliage or with visible symptoms of disease on the leaves lose their ornamental features and become useless as covers for unsightly places in gardens [4,5]. up to now there is no data about the occurrence of micromycetes on climbing roses in botanic garden of the jagiellonian university in cracow. this is why this study was undertaken. doi: 10.5586/am.1054 publication history received: 2014-11-06 accepted: 2015-07-02 published: 2015-08-05 handling editor maria rudawska, institute of dendrology of the polish academy of sciences, poland authors’ contributions mk: concept of the study; kdf, mk: determination of the specimens; mk, kdf: writing the manuscript funding research supported by the polish ministry of science and higher education as part of the statutory activities of the department of plant protection, university of agriculture in krakow. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation kowalik m, duda-franiak k. micromycetes on climbing roses leaves (rosa l.) in the botanic garden of the jagiellonian university in cracow. acta mycol. 2015;50(1):1054. http:// dx.doi.org/10.5586/am.1054 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:m.kowalik%40ogr.ur.krakow.pl?subject=micromycetes%20on%20climbing%20roses%20leaves%20%28rosa%20l.%29%20in%20the%20botanic%20garden%20of%20the%20jagiellonian%20university%20in%20cracow http://dx.doi.org/10.5586/am.1054 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1054 http://dx.doi.org/10.5586/am.1054 2 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1054 kowalik and duda-franiak / micromycetes on leaves of climbing roses material and methods the mycological research was carried out using the collection of climbing roses from the botanic garden of the jagiellonian university in cracow. the source of the research material were leaves of 55 climbing roses bushes, represented by 20 cultivars collected in june, august and october 2011–2013. micromycetes were isolated from 1350 climbing rose leaves. leaf fragments were cut from the border of healthy and necrotic tissues from single spots and surface sterilized in 70% ethanol for one minute, then thoroughly rinsed three times for one minute in sterile water before being placed on a petri dish with a 2% pda medium. the petri dishes were incubated for 7 days at 21–22°c. micromycetes isolation and culture were carried out according to the standard methods practiced in mycology [6]. the microscope used for observation was a delta optical microscope, model evolution 300. the following keys was used in micromycetes taxonomic identification: guba [7], domsch et al. [8], sutton [9], sivanesan [10] and ellis and ellis [11]. the basis for the classification was the system of kirk et al. [12] and the authors’ epithets by fungal species names were verified according to index fungorum 2014 [13]. on the basis of micromycetes specification and considering the participation of individual species in the total fungal community, particular taxa were classified to the group of dominants (constituting of >5% of the entire community), influents (1–5%) and accessory fungi (<1%), according to kowalik [14]. similarity coefficient of the micromycetes communities on rose leaves was calculated including terms and years of research according to the formula given by błaszkowski et al. [15]: s = w / a + b − w, where: s – similarity of compared communities (range of coefficient variation 0–1), a, b – number of species in communities a, b, w – number of shared species in both communities. results community of micromycetes inhabiting leaves of climbing roses from the botanic garden of the jagiellonian university, cracow in 2011–2013 produced differentiated number of colonies and revealed diverse species composition. among one thousand two hundred micromycetes colonies isolated from the infected fragments of climbing roses leaves 65 species have been identified. the number of fungal colonies in the years 2011, 2012 and 2013 amounted 320, 412 and 468 and 27, 45 and 37 taxa has been identified each year respectively. in june, august and october 36 to 181 colonies were isolated, which comprised 8 to 28 species (tab. 1). the dominant (with abundance above 5% of all recovered colonies) micromycetes isolated from leaves of climbing roses comprised the following species (in the descending order): alternaria alternata, epicoccum nigrum, diplocarpon rosae, penicillium brevicompactum and pestalotia rosae. the group of influents (abundance 1–5% of all recovered colonies), included: arthrinium sphaerospermum, aspergillus flavus, a. niger, chaetomium funicola, cladosporium cladosporioides, c. herbarum, humicola grisea, ilyonectria radicicola, mortierella parvispora, penicillim expansum, p. glabrum, p. herquei, pestalotiopsis sydowiana, phialophora cyclaminis, phoma leveillei, ph. medicaginis, pleurostomophora richardsiae and sordaria fimicola. the accessory group, amounting less than 1% of the isolated colonies, included species from the following genera: acremonium, coelophoma, gliomastix, isaria, mucor, nectria, oidiodendron, paraphoma, rhizopus, stemphylium, thanathephorus, trichoderma, trichothecium, umbelopsis and others. aspergillus chevalieri, boeremia exigua, chaetomim cochlioides, ch. piluliferum, humicola fuscoatra, nectria inventa, mucor hiemalis, penicillium citrinum, rhizopus stolonifer, stemphylium vesicarium, trichothecium roseum were occasionally found in the phyllosphere of roses as single or double colonies. alternaria alternata and c. cladosporioides constantly inhabited leaves with necrosis symptoms, although with a different frequency. the pathogen d. rosae appeared continuously on the leaves (except year 2011), while p. rosae was found from august to october, with a greater number of colonies recorded in october. the 3 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1054 kowalik and duda-franiak / micromycetes on leaves of climbing roses ta b. 1 m ic ro m yc et es is ol at ed fr om th e in fe ct ed le av es o f c lim bi ng ro se s (r os a l. ). fu ng us fu ng i f re qu en cy o n le av es in : to ta l % 20 11 20 12 20 13 ju ne a ug us t o ct ob er ju ne a ug us t o ct ob er ju ne a ug us t o ct ob er a cr em on iu m r ut ilu m w . g am s 1 2 1 1 5 0. 42 a lte rn ar ia a lte rn at a (f r.) k ei ss l. 16 20 30 24 62 64 62 17 40 33 5 27 .9 2 a lte rn ar ia b ot ry tis (p re us s) w ou de nb er g & c ro us 2 2 0. 17 a rt hr in iu m sp ha er os pe rm um f uc ke l 7 9 16 1. 33 a sp er gi llu s c he va lie ri th om & c hu rc h 1 1 0. 08 a sp er gi llu s fl av us l in k 3 1 5 5 14 1. 17 a sp er gi llu s n ig er th ie gh . 1 8 7 16 1. 33 a sp er gi llu s s yd ow ii (b ai ni er & s ar to ry ) th om & c hu rc h 4 1 1 6 0. 50 a sp er gi llu s v er si co lo r (v ui ll. ) t ir ab . 2 2 2 6 0. 50 bo er em ia e xi gu a (d es m .) a ve sk am p, g ru yt er & v er kl ey 1 1 0. 08 bo tr yt is c in er ea p er s. 5 4 1 10 0. 83 c ha et om iu m c oc hl io de s p al lis er 1 1 0. 08 c ha et om iu m fu ni co la c oo ke 4 3 2 2 2 13 1. 08 c ha et om iu m g lo bo su m k un ze 2 1 2 5 0. 42 c ha et om iu m p ilu lif er um j. d an ie ls 2 2 0. 17 c la do sp or iu m c la do sp or io id es (f re se n. ) g .a d e v ri es 1 1 2 5 14 6 4 5 5 43 3. 58 c la do sp or iu m h er ba ru m (p er s. ) l in k 2 5 1 4 9 5 3 29 2. 42 c la do sp or iu m sp ha er os pe rm um p en z. 3 2 5 0. 42 c ol eo ph om a em pe tr i ( r os tr .) d ie d. 1 3 4 0. 33 d ip lo ca rp on ro sa e f. a . w ol f 10 21 3 1 2 1 21 3 62 5. 17 ep ic oc cu m n ig ru m l in k 4 10 49 1 4 1 9 78 6. 50 4 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1054 kowalik and duda-franiak / micromycetes on leaves of climbing roses ta b. 1 c on tin ue d fu ng us fu ng i f re qu en cy o n le av es in : to ta l % 20 11 20 12 20 13 ju ne a ug us t o ct ob er ju ne a ug us t o ct ob er ju ne a ug us t o ct ob er g lio m as tix lu zu la e (f uc ke l) e .w . m as on e x s. h ug he s 2 3 1 6 0. 50 h um ic ol a fu sc oa tr a tr aa en 1 1 0. 08 h um ic ol a gr is ea t ra ae n 1 1 3 4 7 16 1. 33 ily on ec tr ia ra di ci co la (g er la ch & l . n ils on ) p. c ha ve rr i & c . s al ga do 8 10 10 28 2. 33 is ar ia fu m os or os ea w iz e 7 1 8 0. 67 k hu sk ia o ry za e h .j. h ud s. 1 4 5 0. 42 m or tie re lla a lp in a pe yr on el 2 1 2 5 0. 42 m or tie re lla h ya lin a (h ar z) w . g am s 9 9 0. 75 m or tie re lla p ar vi sp or a li nn em . 3 5 5 2 8 1 4 28 2. 33 m uc or h ie m al is w eh m er 1 1 0. 08 m yr ot he ci um v er ru ca ri a (a lb . & s ch w ei n. ) d itm ar 3 3 0. 25 n ec tr ia in ve nt a pe th yb r. 1 1 0. 08 o id io de nd ro n ec hi nu la tu m g .l . b ar ro n 7 7 0. 58 pa ra co ni ot hy ri um m in ita ns (w .a . c am pb .) v er kl ey 6 2 8 0. 67 pa ra ph om a ch ry sa nt he m ic ol a (h ol ló s) g ru yt er , a ve sk am p & v er kl ey 2 2 4 0. 33 pe ni ci liu m d od ge i p itt 3 3 0. 25 pe ni ci lli um b re vi co m pa ct um d ie rc kx 13 29 19 61 5. 08 pe ni ci lli um c itr in um th om 1 1 0. 08 pe ni ci lli um e xp an su m l in k 10 9 19 1. 58 pe ni ci lli um g la br um (w eh m er ) w es tli ng 2 9 16 17 44 3. 67 5 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1054 kowalik and duda-franiak / micromycetes on leaves of climbing roses ta b. 1 c on tin ue d fu ng us fu ng i f re qu en cy o n le av es in : to ta l % 20 11 20 12 20 13 ju ne a ug us t o ct ob er ju ne a ug us t o ct ob er ju ne a ug us t o ct ob er pe ni ci lli um h er qu ei b ai ni er & s ar to ry 6 15 8 29 2. 42 pe ni ci lli um ja va ni cu m j. f. h . b ey m a 1 1 0. 08 pe ni ci lli um je ns en ii k .m . z al es sk y 1 3 4 0. 33 pe ni ci lli um m ic zy ns ki k .m . z al es sk y 2 1 3 0. 25 pe ni ci lli um re st ri ct um j. c . g ilm an & e .v . a bb ot t 1 7 8 0. 67 pe ri co ni a m ac ro sp in os a le fe bv re & jo hn so n 4 4 0. 33 pe st al ot ia ro sa e w es te nd . 8 12 7 20 2 2 10 61 5. 08 pe st al ot io ps is sy do w ia na (b re s. ) b . s ut to n 13 1 14 1. 17 ph ia lo ph or a cy cl am in is j. f. h . b ey m a 10 4 14 1. 17 ph om a co m pl an at a (t od e) d es m . 3 3 0. 25 ph om a eu py re na s ac c. 2 4 1 7 0. 58 ph om a le ve ill ei b oe re m a & g .j. b ol le n 3 5 5 2 4 19 38 3. 17 ph om a m ed ic ag in is m al br & r ou m . 8 3 1 12 1. 00 ph om a pu ta m in um h ol ló s 2 2 0. 17 pl eu ro st om op ho ra r ic ha rd si ae (n an nf .) l. m os re rt , w . g am s & c ro us 4 11 5 20 1. 67 r hi zo pu s s to lo ni fe r (e hr en b. ) v ui ll. 1 1 0. 08 so rd ar ia fi m ic ol a (r ob er ge e x d es m .) c es . & d e n ot 9 11 12 4 6 6 48 4. 00 st em ph yl iu m v es ic ar iu m (w al lr. ) e .g . s im m on s 1 1 0. 08 th an at ep ho ru s c uc um er is (a .b . f ra nk ) d on k 2 2 0. 17 tr ic ho de rm a vi ri de p er s. 5 1 6 0. 50 tr ic ho th ec iu m ro se um (p er s. ) l in k 1 1 2 0. 17 6 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1054 kowalik and duda-franiak / micromycetes on leaves of climbing roses necrotroph e. nigrum colonized massively the leaves of climbing roses in october 2011. species of cladosporium and phoma, including c. cladosporioides, c. herbarum, c. sphaerospermum, ph. companata, ph. eupyrena, ph. leveillei, ph. medicaginis and ph. putaminum, inhabited roses leaves in large numbers. the similarity between two of the compared communities (within three terms for each year of the study and between the same terms) ranged from 0.13 to 0.60 (tab. 2). in the growing season 2011 the similarity indices ranged from 0.29 to 0.38, in 2012 from 0.21 to 0.31, and in 2013 from 0.44 to 0.6. the lowest similarity coefficient, calculated for the micromycetes communities on the leaves of climbing roses was found in june 2011 and august 2012, and the highest was for the community found in june and august 2013. eleven compared communities were characterized by very low similarity coefficients (in the range 0.1 to 0.2), 15 communities were characterized by low coefficient (0.21–0.3), seven communities as middle coefficient (0.31–0.4), two as high coefficient (0.41–0.5), and only a single community as the highest (0.51–0.60). discussion the phyllosphere of climbing roses was dominated by fungus a. alternata for three consecutive years. this polyphagous species has been mentioned in many studies as the cause of extensive necrosis on leaves [6,14,16,17]. scheffer [18] speculates that this may be the most widespread species in the world, both in temperate and tropical climates. it is considered to be a very persistent species because its dried spores are able to survive for a years [8]. dominance of a. alternata in community structure of micromycetes on the leaves of climbing roses proved to be particularly harmful and as a consequence caused numerous necrosis on leaves. this fungus can also colonize the dead tissue of rose leaves that was previously necrotized by pathogens. therefore in this study we collected fragments from the edges of the living and dead leaf tissue of roses. such approach excludes the isolation of endophytes because they inhabit the asymptomatic tissue of their hosts. the high frequency of a. alternata on the leaves of roses, confirmed by the isolation of 335 colonies, favored a significant level of infectivity and pathogenicity of this necrotroph. the proximity of many host plants also created favorable conditions for its spread, causing increased colonization and infection, as documented in earlier studies [2,14,17]. the production of host specific toxins had an impact on dieback and premature leaf fall, showing its toxic properties in relation to host plants, which is emphasized by wakuliński [19]. previously duda and bonio [16] and kowalik et al. [17,20,21] have documented the high frequency of a. alternata and other potentially pathogenic micromycetes in the botanic garden of the jagiellonian university in cracow. the most frequently isolated species from the leaves of lamiaceae family herbs, azalea pontica and saucer ta b. 1 c on tin ue d fu ng us fu ng i f re qu en cy o n le av es in : to ta l % 20 11 20 12 20 13 ju ne a ug us t o ct ob er ju ne a ug us t o ct ob er ju ne a ug us t o ct ob er u lo cl ad iu m c on so rt ia le (th üm ) e .g . s im m on s 2 2 0. 17 u m be lo ps is is ab el lin a (o ud em .) w . g am s 1 3 1 5 0. 42 u m be lo ps is v in ac ea (d ix on -s te w .) a rx 1 1 0. 08 to ta l c ol on ie s 36 11 9 16 5 87 14 4 18 1 15 5 14 9 16 4 12 00 10 0. 0 to ta l s pe ci es 8 19 17 20 18 28 25 23 24 65 to ta l c ol on ie s in y ea r 32 0 41 2 46 8 12 00 to ta l s pe ci es in y ea r 27 45 37 65 7 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1054 kowalik and duda-franiak / micromycetes on leaves of climbing roses magnolia were a. alternata, e. nigrum, s. fimicola, p. expansum and pestalotiopsis sydowiana. the colonization of leaves of roses by fungal species found on the other plant species grown in the vicinity of rosaceous plants indicates on the strong liaison of pathogenic fungi and saprobes in phyllospheres of different plants. according to jędryczka [22], the alternaria and cladosporium genera are particularly frequent in the air, and the aspergillus and penicillium genera are often listed as aerosols [23]. bonio and duda [23], evaluating the number of colony forming units in the botanic garden of the jagiellonian university in cracow and the rogów arboretum of the warsaw university of life sciences, reported that a. alternata was the dominant fungus in the air near these two locations. ogórek et al. [24] indicated that a. alternata is a ubiquitous fungus with a very high concentration of spores in the air of mountain areas. cladosporium cladosporioides, c. herbarum and c. sphaerospermum inhabit the leaves of climbing roses quite frequently. jędryczka [22] states that their presence in the atmospheric air in the form of bioaerosols favors their infectious potential, causing necrosis on colonized plant tissue [25,26]. the weather and local climate can cause an increase density of alternaria, cladosporium and epicoccum spores in the air [23]. these studies may suggest that fungi mentioned above colonize the leaves of plants under conditions of slight moisture. a somewhat higher colonization of leaves of roses in october than in the summer months by fungi from the genus mortierella is probably related to their preferences in terms of higher humidity. more than 60 colonies of p. rosae were isolated in the present study. this micromycete is monophagous and was found on roses in the vicinity of ghent and described for the first time in 1859 [7]. in polish literature this fungus has not been described up to now. according to mycological analyses, the most frequent disease occurring on climbing roses leaves was rose black spot caused by d. rosae, which resulted in dieback and drooping of leaves in june for three consecutive years of the study (particularly evident in 2011). symptoms of grey mould caused by b. cinerea were visible on the leaves until june 2013, which confirms the opinion that the pathogen is not dangerous for strong, wellfed shrubs [2]. the number of isolated micromycetes colonies was almost five times higher in october 2011 than in june of 2011, twice as high in october 2012 than in june 2012 and similar in both terms of 2013. this reflects the increasing infestation towards the end of the growing season that appears as spots and leaf necrosis, resulting in defoliation [5]. the increase of the number of colonies and species of micromycetes obtained in the third year of the study may indicate on the accumulation of micromycetes propagules on tested climbing roses. tab. 2 similarity coefficient between micromycetes communities isolated from the leaves of climbing roses (rosa l.). year 2011 2012 2013 month vi viii x vi viii x vi viii x 20 11 vi 0.29 0.32 0.27 0.13 0.20 0.14 0.19 0.19 viii 0.29 0.38 0.30 0.23 0.34 0.19 0.20 0.23 x 0.32 0.38 0.32 0.25 0.36 0.24 0.29 0.28 20 12 vi 0.27 0.30 0.32 0.31 0.26 0.18 0.16 0.19 viii 0.13 0.23 0.25 0.31 0.21 0.26 0.24 0.17 x 0.20 0.34 0.36 0.26 0.21 0.33 0.28 0.30 20 13 vi 0.14 0.19 0.24 0.18 0.26 0.33 0.60 0.44 viii 0.19 0.20 0.29 0.16 0.24 0.28 0.60 0.47 x 0.19 0.23 0.28 0.19 0.17 0.30 0.44 0.47 8 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1054 kowalik and duda-franiak / micromycetes on leaves of climbing roses conclusions (i) communities of micromycetes inhabiting the leaves of climbing roses in the botanic garden of the jagiellonian university in cracow were characterized by different species composition in the three successive years of research. (ii) the dieback of climbing rose leaves was caused by numerous micromycetes. the dominant role was played by the pathogen d. rosae and necrotrophs a. alternata, e. nigrum, p. brevicompactum and s. fimicola. references 1. wiśniewska-grzeszkiewicz h. róże w ogrodzie. warszawa: multico; 2009. 2. kowalik m, pokora b, chimowska b. róże w chorzowskim rosarium, ich patogeny a człowiek. in: wiech k, kołoczek h, kaszycki p, editors. ochrona środowiska naturalnego w xxi wieku – nowe wyzwania i zagrożenia. kraków: fundacja na rzecz wspierania badań naukowych wydziału ogrodniczego akademii rolniczej im. h. kołłątaja w krakowie; 2005. p. 86–94. 3. wojdyła a, kamińska m, łabanowski g, orlikowski l. ochrona róż. kraków: wydawnictwo plantpress, sp. z o.o.; 2002. 4. pusz w. podstawy ochrony roślin w ogrodach przydomowych i na działkach. warszawa: oficyna wydawnicza “hoża” spółdzielnia pracy; 2011. 5. pusz w. plamistości liści. mój piękny ogród. 2012;10:41. 6. kowalik m. fungi and fungi-like oomycetes isolated from affected leaves of rhododendron. acta mycol. 2008;43(1):21–27. http://dx.doi.org/10.5586/am.2008.003 7. guba ef. monograph of monochaetia and pestalotia. cambridge, ma: harvard university press; 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[1] there are 980 aphyllophoroid species found in finland. aphyllophoroid fungi are important in decomposition of dead wood, and, thus they play an important role in woody ecosystems. in addition to wood decayers, a part of aphyllophoroid fungi are litter decayers, ectomycorrhizal, parasitic fungi and mycoparasites [1]. original research paper acta mycol 49(2):221–235 doi: 10.5586/am.2014.017 received: 2014-06-01 accepted: 2014-10-15 published electronically: 2014-12-31 noteworthy records of aphyllophoroid fungi in finland (basidiomycota) panu kunttu1*, jorma pennanen2, tapio kekki3, matti kulju4, mai suominen1 1 school of forest sciences, university of eastern finland, p.o. box 111, 80101 joensuu, finland 2 pentbyntie 1 a 2, 10300 karjaa, finland 3 kiirunankatu 8, 96400 rovaniemi, finland 4 ylispuuntie 13, 90420 oulu, finland abstract we present new records of noteworthy aphyllophoroid fungi, mainly polypores and corticioids in finland. the following 19 rare or infrequently collected species are presented with notes on their substrates: amylocorticium subsulphureum, antrodiella parasitica, ceraceomyces sulphurinus, clavaria atroumbrina, clavaria rosea, gloeophyllum carbonarium, hyphodontia flavipora, junghuhnia fimbriatella, lindtneria chordulata, odonticium septocystidia, peniophorella guttulifera, perenniporia tenuis, postia immitis, repetobasidium vile, resinicium pinicola, sidera vulgaris, tomentella coerulea, trechispora laevis and xylodon pruni. we also list 41 aphyllophoroid fungi as new to some sections of the boreal vegetation zone in finland. keywords: biogeography; boreal forest; corticioid; distribution; polypore; rare species * corresponding author. email: panu.kunttu@uef.fi handling editor: tomasz leski http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.2014.017 mailto:panu.kunttu%40uef.fi?subject=am.2014.017 222© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) so far, there are only a few intensive inventories done in finland that cover all aphyllophorales. especially other aphyllophoroid fungi than polypores are time consuming to sample: for example basidiocarps of corticioids are small in size and often they grow underneath a trunk. therefore they are demanding to find, and most of these species need to be identified with a microscope. because of low sampling effort, most of these species are poorly known. the first finnish checklist of aphyllophoroid fungi was published in 2009 [1]. after this, plenty of biogeographical knowledge has accumulated especially about polypores and corticioids, and new records have been published for example by kunttu et al. [2–6], kotiranta and larsson [7], kotiranta and shiryaev [8] and spirin et al. [9]. in this article we present new records of aphyllophoroid fungi mainly from two larger field works that were conducted in finland in the southern archipelago and in northern lapland, but also some sporadic records is included. we consider two kinds of records: rare or little collected species with five earlier records in finland and species that are new to some section (subzone) of boreal vegetation zone in finland. material and methods the records presented in this article come from several inventories in different parts of finland. records are mainly made during a period from 2008 to 2013, but some older records are also included due to change of the taxonomical status. these studies are mostly connected to inventories of species assemblages in certain areas or ecological research of polypores and corticioids. most of the records come from inventories made by metsähallitus, natural heritage services. these records are from nature conservation areas where the purpose of the inventory has been to map species living in these areas. the inventory methods have been various. records are also made in an ecological and biogeographical study that concentrates on polypores and corticioids in the southwestern archipelago of finland (p. kunttu, doctoral dissertation in preparation). some records are made during xxi nordic mycological congress in rovaniemi in september 2013. additional records are also made on field trips by authors during their leisure time. we use finnish national uniform grid system (27°e) according to heikinheimo and raatikainen [10] for coordinates and the abbreviation ucs refers to uniform coordinate system. biogeographical provinces are according to knudsen and vesterholt [11]. boreal and hemiboreal vegetation zones consist of 11 sections (subzones), which are represented in this article (fig. 1) [12,13]. voucher specimens are deposited in the herbaria of the universities of helsinki (h), turku (tur), oulu (oulu), slovak national museum (bra) and/or in the reference herbarium of jorma pennanen (j.p.), kaisa junninen (k.j.), matti kulju (m.k.) or heikki kotiranta (h.kta). nomenclature is mainly according to kotiranta et al. [1], but names of some species follow miettinen and larsson [14] and bernicchia and gorjón [15]. nomenclature of the genus hyphodontia sensu lato follows hjortstam and ryvarden [16]. the finnish national red-listing evaluation of iucn standard is according to kotiranta et al. [17]. the decay stage classification of trunks (1–5) is according to renvall [18]. diameter of a 223© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) trunk is given at the breast height if the trunk has been unbroken and at the base if the trunk has been broken. material has been collected, identified or confirmed by several researches that are mentioned in the collecting information. at least five times are mentioned the following researchers: tk = tapio kekki, hk = heikki kotiranta, mk = matti kulju, pk = panu kunttu, jp = jorma pennanen. collector is also the identifier if not otherwise stated. number code after the collectors name or abbreviation of the collector is a personal sampling number of the specimen. fig. 1 boreal and hemiboreal vegetation zones and their sections in finland. 224© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) results we present new records of following 19 rare or little collected species and give notes of their substrates. these species are amylocorticium subsulphureum, antrodiella parasitica, ceraceomyces sulphurinus, clavaria atroumbrina, clavaria rosea, gloeophyllum carbonarium, hyphodontia flavipora, junghuhnia fimbriatella, lindtneria chordulata, odonticium septocystidia, peniophorella guttulifera, perenniporia tenuis, postia immitis, repetobasidium vile, resinicium pinicola, sidera vulgaris, tomentella coerulea, trechispora laevis and xylodon pruni. we also list 41 aphyllophoroid fungi as new to some section (subzone) of the boreal vegetation zone in finland. species are presented in an alphabetic order. the number of new species to some section (subzone) of boreal vegetation zone are as follows: hemiboreal, oak zone (1b) 2; southern boreal, southwestern finland and southern ostrobothnia (2a) 2; southern boreal, lake district (2b) 2; middle boreal, ostrobothnia (3a) 1; middle boreal, northern carelia – kainuu (3b) 4; middle boreal, southwestern lapland (3c) 11; northern boreal, north ostrobothnia (4b) 11; northern boreal, forest lapland (4c) 8. there are 1 critically endangered, 4 endangered, 3 vulnerable, 4 near threatened and 3 data deficient species among the taxa presented here. list of species amylocorticium subsulphureum (p. karst.) pouzar specimen examined: savonia borealis, heinävesi, kermajärvi, ucs 6929959:3587206, 17.ix.2008, jp ss42/2 (h + j.p.), on picea abies, whole fallen trunk, diam. 30 cm, decay stage 3. near threatened. new to southern boreal, lake district (2b). this is the fourth record in finland [1]. anomoloma myceliosum (peck) niemelä & k.h. larss. specimen examined: karelia borealis, lieksa, möhkyrinkangas, ucs 7033531:3685196, autumn 2011, mari e. niemi 143 (tur), clear cut area with 50 m3/ha dead wood, advanced picea abies trunk, diam. 25 cm. near threatened. new to middle boreal, northern carelia – kainuu (3b). antrodia sitchensis (baxter) gilb. & ryvarden specimens examined: savonia borealis, heinävesi, kermajärvi, ucs 6924419:3591868, 24.ix.2008, jp hs158/4 (h + j.p.), on picea abies, whole fallen trunk, diam. 45 cm, decay stage 3. oxalis-myrtillus forest type; ostrobottnia ultima, rovaniemi, hyypiökivalo ucs 7358676:3487201, 11.viii.2009, juha kinnunen 5194 (h), on picea abies, whole fallen trunk, diam. 40 cm, decay stage 2; savonia borealis, savonranta, kakonsalo, haukiniemi, ucs 6910310:3605339, 15.ix.2009, jp 964 (h + j.p.), on pinus sylvestris, broken fallen trunk, diam. 45 cm, decay stage 3, myrtillus type forest.; ostrobottnia kajanensis, sotkamo, talvivaara, ucs 7092700:3556644, 25.viii.2010, anni markkanen 2718 (jyv), on picea abies, whole fallen trunk, diam. 20 cm, decay stage 3; karelia borealis, lieksa, ukonsärkkä, ucs 7032840:3689321, 22.ix.2010, kaisa junninen 7805 (h), on picea abies, whole fallen trunk, diam. 35 cm, decay stage 3. endangered. new to northern boreal, north ostrobothnia (4b). 225© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) antrodiella parasitica vampola specimens examined: nylandia, sipoo, rörstrand, ucs 6706981:3400127, 15.x.2008, keijo savola h/151008, det. mari e. niemi (h), on picea abies, whole fallen trunk, diam. 28 cm, decay stage 3; nylandia, sipoo, rörstrand, ucs 6706479: 3400459, 15.x.2008, keijo savola u/151008, det. mari e. niemi (h), on picea abies, whole fallen trunk, diam. 37 cm, decay stage 4. vulnerable. this is the sixth occurrence in finland [1]. bjerkandera fumosa (pers. : fr.) p. karst. specimen examined: karelia borealis, lieksa, haapahaasianvaara, ucs 7031313:3687422, 19.ix.2008, kaisa junninen 7619, det. tuomo niemelä (k.j.), on populus tremula, whole fallen trunk, diam. 37 cm, decay stage 2. new to middle boreal, northern carelia – kainuu (3b). bulbillomyces farinosus (bres.) jülich specimen examined: lapponia kittilensis, muonio, pallas-ounastunturi national park, pallastunturi s, ucs 7553415:3376871, 27. viii. 2013, jp 2565 (h, j.p.), on betula sp. fallen trunk, diam. 15 cm decay stage 2, old hylocomium-myrtillus type forest. new to northern boreal, forest lapland (4c). calocera cornea (batsch : fr.) fr. specimen examined: ostrobottnia ultima, rovaniemi, pisajärvi nature reserve, ucs 7362840:3418095, 2.ix.2011, tk 297 (h), fallen trunk of betula sp. in herb rich forest. new to middle boreal, southwestern lapland (3c). ceraceomyces sulphurinus (p. karst.) j. erikss. & ryvarden specimen examined: tavastia borealis, savonranta, kakonsalo, aluslamminmäki, ucs 6908677:3601099, 10.ix.2009, jp 910 (h, j.p), on populus tremula, fallen trunk, diam 50 cm, decay stage 4. fruitbody grew on the bark and its length was few meters. habitat was an old lakeside mixed forest. vulnerable. this is the third record in finland. earlier records are from punkaharju 1960 (savonia australis) and jalasjärvi 1859 (ostrobottnia australis) [1]. in europe c. sulphurinus has been found in germany, netherlands, belgium, italy, portugal, spain and belarus [15]. ceraceomyces violascens (fr.: fr.) jülich specimen examined: lapponia enontekiensis, enontekiö, pallas-ounastunturi national park, ounastunturi s, ucs 7575152:3374884, 15.viii.2013, jp 2459, conf. mk (h, j.p.), on betula sp., fallen trunk, diam. 15 cm, decay stage 4, mixed birch dominated forest. new to northern boreal, forest lapland (4c). ceriporiopsis aneirina (sommerf.) domanski specimen examined: lapponia kittilensis, kolari, pallas-ounastunturi national park, pyhätunturi nw, ucs 7513940:3383478, 2. ix. 2013, jp 2623 (h, j.p.), on populus tremula fallen trunk, diam. 30 cm, decay stage 3, mixed spruce dominated forest. near threatened. new to northern boreal, north ostrobothnia (4b). 226© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) clavaria asterospora pat. sensu lato specimen examined: ostrobottnia ultima, tervola, kätkävaara, ucs 7348244:3406879, 2.ix.2012, tk 580 (tur), on track in calciferous picea abies dominated forest. this is distinct from clavaria falcata with ellipsoid spores. new to middle boreal, southwestern lapland (3c). clavaria atroumbrina corner specimen examined: ostrobottnia ultima, keminmaa, kallinkangas, ucs 7304:3387, 5.ix.2013, viktor kučera (bra cr19576), det. i. kautmanová, on calciferous ground in graveyard. new to middle boreal, southwestern lapland (3c). one record of clavaria cf. pullei donk has been made in kuhmoinen (tavastia australis), which is most probably a synonym for c. atroumbrina. so this would make it the second record in finland [1]. clavaria fragilis holmsk. : fr. specimen examined: ostrobottnia ultima, rovaniemi, kiiruna, ucs 737876:344484, 10.viii.2010, tk 31 (tur), in sandy yard with antennaria dioica. new to northern boreal, north ostrobothnia (4b). clavaria rosea dalman specimen examined: ostrobottnia media, kokkola, halkokari, ucs 708918:331036, 19.ix.2011, terho taarna (oulu) conf. esteri ohenoja, lawn in garden; savonia australis, kerimäki, ucs 686949:362045, 25.vii.2012, tk 554 (tur), lawn in garden; savonia australis, kerimäki, louhi, ucs 687186:360643, 17.viii.2012, mauri lahti 28/12 (tur), lawn near lime quarry. new to middle boreal, ostrobothnia (3a). there are three earlier records made in finland: two from saari (karelia ladogensis) and one from kuusamo (regio kuusamoënsis) [1]. clavicorona taxophila (thom) doty specimen examined: ostrobottnia ultima, rovaniemi, pisajärvi, ucs 7362858:3418110, 2.ix.2011, tk 298 (tur), mesic herb-rich forest. new to middle boreal, southwestern lapland (3c). clavulina rugosa (bull. : fr.) j. schröt. sensu lato specimen examined: ostrobottnia ultima, rovaniemi, savioja, ucs 734902:342663, 23.ix.2011, tk 498 (tur), moist herb-rich forest. new to middle boreal, southwestern lapland (3c). gloeophyllum carbonarium (berk & m.a. curtis) ryvarden specimens examined: karelia borealis, ilomantsi, patvinsuo, ucs 6999276:3690969, 2.ix.2008, kaisa junninen 7492 (h), on pinus sylvestris, fallen broken trunk, charred, diam. 17 cm, decay stage 2; tavastia borealis, savonranta, kakonsalo, raatelamminsalo, ucs 6908478:3602133, 24.ix.2009, jp 1039 (h, j.p.), on pinus sylvestris, charred rootstalk, diam. 30 cm, decay stage 3. endangered. according to kotiranta et al. [1] there are five earlier records in finland but there are several unpublished records made in ostrobottnia kajanensis (t. helo, personal communication, 2014). 227© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) hypochnicium bombycinum (sommerf. & fr.) j. erikss. specimen examined: lapponia enontekiensis, enontekiö, pallas-ounastunturi national park, röyninkuru, ucs 7565306:3378545, 28.viii.2013, jp 2574, conf. mk (h, j.p.), on living salix sp., diam. 10 cm, old and moist, mixed spruce dominated forest. new to northern boreal, forest lapland (4c). hyphodontia flavipora (cooke) sheng h. wu specimen examined: regio aboënsis, kemiönsaari, dragsfjärd, vänö, ucs 664:323, 15.xi.2008, timo kosonen 737-2008, det. jp (tur), on alnus glutinosa, fallen trunk, diam. 6 cm, decay stage 2. data deficient. there is one earlier record made in finland: naantali (regio aboënsis) in 1997 [1]. fruitbody grew on betula sp. junghuhnia fimbriatella (peck) ryvarden savonia borealis, tavastia borealis, savonranta, kakonsalo, aluslamminmäki, ucs 6908721:3601130, 10.ix.2009, jp 912 (h, j.p.), on populus tremula, fallen whole trunk, diam. 50 cm, decay stage 3. j. fimbriatella grew on dead fruitbody of ganoderma applanatum. this is the second record in finland. according to niemelä [19] the first one was found from joensuu in 2006 (karelia borealis). in europe j. fimbriatella has been found in germany, poland, switzerland, austria, czech republic, former yugoslavia area and russia. in addition to these there are records from kamchatka peninsula, canada and united states. the substrates have been populus, fagus and fraxinus [15]. kneiffiella alienata (s. lundell) jülich & stalpers specimen examined: lapponia kittilensis, muonio, pallas-ounastunturi national park, pyhäkero e, ucs 7551549:3381223, 20.viii.2013, jp 2490, conf. hk (h, h.kta, j.p.), on picea abies fallen trunk, diam. 20 cm, decay stage 4, moist herb-rich forest. new to northern boreal, forest lapland (4c). kneiffiella barba-jovis (bull.) p. karst. specimen examined: lapponia enontekiensis, enontekiö, pallas-ounastunturi national park, ounastunturi s, ucs 7577820:3375140, 13.viii.2013, jp 2433 (h, j.p.), on betula sp., fallen branch, diam. 5 cm, decay stage 3, pine dominated sub-xeric heath forest. new to northern boreal, forest lapland (4c). kneiffiella subalutacea (p. karst.) jülich & stalpers specimen examined: lapponia kittilensis, kolari, pallas-ounastunturi national park, pyhätunturi nw, ucs 7514455:3382142, 2.ix.2013, jp 2615 (h, j.p.), on pinus sylvestris, fallen trunk, diam. 25 cm, decay stage 3, mixed pine dominated forest. new to northern boreal, north ostrobothnia (4b). lentaria afflata (lagger) corner specimen examined: ostrobottnia ultima, keminmaa, kallinkangas nature reserve, ucs 7303990:3386811, 12.ix.2012, tk 691 (tur), on fallen trunk of populus tremula, calciferous herb-rich heath forest. new to middle boreal, southwestern lapland (3c). 228© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) lindtneria chordulata (d.p. rogers) hjortstam specimens examined: regio aboënsis, kemiönsaari, dragsfjärd, yxskär, ucs 6650562:3223065, 5.viii.2010, pk 6494, det. mk (tur), on populus tremula, fallen broken trunk, diam. 27, decay stage 4; regio aboënsis, kemiönsaari, dragsfjärd, storlandet, apelholmen, ucs 6653:3247, 18.viii.2010, pk 6709, det. mk (tur), on populus tremula, fallen whole trunk, diam. 22 cm, decay stage 1. vulnerable. six records are made in finland before: helsinki in four places (nylandia) on salix caprea, salix sp., coniferous board, pinus timber and syringa vulgaris (cult.) [1]; lam-mi (tavastia australis) on corylus [1], parainen, nauvo on populus tremula (regio aboënsis) [5]. macrotyphyla fistulosa (holmsk. : fr.) r.h. petersen specimen examined: ostrobottnia ultima, rovaniemi, marrasjärvi, ucs 741979:342391, 16.x.2011, tk 531 (tur), on fallen branch of betula pendula, diam. 1, xeric heath forest. specimen was m. fistulosa var. contorta (holmsk.) nannf. & l. holm. new to northern boreal, north ostrobothnia (4b). mucronella bresadolae (quél) corner ostrobottnia ultima, rovaniemi, pisavaara strict nature reserve, ucs 7359279:3416987, 12.ix.2013, mk 12/13 & pekka helo (oulu), on pinus sylvestris. new to middle boreal, southwestern lapland (3c). odonticium septocystidia (burt) zmitr. & spirin (fig. 2) fig. 2 odonticium septocystidia, regio aboënsis, salo, kisko, pappilanniemi, 11.x.2013, jp 2745. photo: jorma pennanen. 229© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) specimens examined: regio aboënsis, salo, kisko, pappilanniemi nature reserve, ucs 6687518:3303916, 11.x.2013, jp 2740, conf. hk (h, h.kta, j.p.), on decorticated populus tremula fallen branch, diam. 20 cm, decay stage 2, herb-rich heath forest; regio aboënsis, salo, kisko, pappilanniemi nature reserve, ucs 6687621:3304145, 11.x.2013, jp 2745, conf. hk (h, h.kta, j.p., m.k.), on decorticated populus tremula fallen trunk, diam. 12 cm, decay stage 3, herb-rich heath forest. endangered. new to hemiboreal, oak zone (1b). this is the second occurrence (on two trunks near each other) in finland, the first was found from tavastia australis, lammi, kotinen nature reserve in 2001 [1]. also in this case the fruiting body grew on decorticated populus tremula [1]. it seems to be rare and in europe the distribution covers germany, estonia, france, belgium, united kingdom, denmark, norway, finland, switzerland and caucasus [15]. in addition to these records it has been found from north america [20]. peniophorella guttulifera (p. karst.) k.h. larss. specimen examined: regio aboënsis, parainen, korppoo, wattkast, söderviken, ucs 6687234:3202197, pk 8179, det. mk (tur), 9.x.2013, betula sp. fallen trunk, diam. 9 cm, decay stage 2. near threatened. according to kotiranta et al. [1] there are records from helsinki 1988–1990 and vantaa 1993 (nylandia), siikainen 1939 (satakunta), tammela 1888 (tavastia australis) and pieksänmaa 2006 (savonia borealis). perenniporia tenuis (schwein.) ryvarden specimen examined: karelia borealis, lieksa, jeremianvaara, ucs 7026980:3686433, october 2011, olli manninen (tur), clear cut area with 50 m3/ha partly fallen retention trees, advanced decayed trunk of populus tremula, diam. 23 cm. critically endangered. there are five earlier records in finland [1]. phanerochaete calotricha (p. karst.) j. erikss. & ryvarden specimen examined: lapponia enontekiensis, enontekiö, pallas-ounastunturi national park, röyninkuru, ucs 7565649:3378816, jp 2580 (h, j.p.), conf. hk, 28.viii.2013, on sorbus aucuparia fallen trunk, diam. 5 cm, decay stage 3, old and moist, mixed spruce dominated forest. new to northern boreal, forest lapland (4c). piloderma olivaceum (parmasto) hjortstam specimen examined: regio aboënsis, perniö, arpalahti, kaapinmäki, ucs 669254:328513, 13.x.2003, maija-liisa & pekka heinonen 850-2003, det. mk (tur), on pinus sylvestris; satakunta, säkylä, iso-säkylä, ucs 6778:3255, 16.x.2001, maija-liisa & pekka heinonen 842-2001, det. mk (tur), on pinus sylvestris; tavastia australis, somero, palikainen, ucs 672402:331858, 26.x.1998 maija-liisa & pekka heinonen 952-1998, det. mk (tur), on pinus sylvestris; tavastia australis, ruovesi, susimäki nature reserve, ucs 686426:335466, 14.ix.1999 maija-liisa & pekka heinonen 685-1999, det. mk (tur), on pinus sylvestris; karelia borealis, lieksa, säynäsemä, ucs 7038:3683, 23.viii.1999 maarit similä & mari niemi 501/1999, det. mk (tur), on pinus sylvestris; karelia borealis, lieksa, vilponkangas, ucs 7024:3691, 16.viii.1999 maarit similä & mari niemi 150/1999, det. mk (tur), on pinus sylvestris; ostrobottnia ultima, rovaniemi, pisavaara strict nature reserve, ucs 7359276:3416918, 12.ix.2013, mk 15/13 & pekka helo (oulu), on pinus sylvestris. 230© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) new to hemiboreal, oak zone (1b), southern boreal, southwestern finland and southern ostrobothnia (2a), southern boreal, lake district (2b), middle boreal, northern carelia – kainuu (3b), middle boreal, southwestern lapland (3c). kotiranta et al. [1] presents only one record from finland, but after that many records p. olivaceum have emerged (h. kotiranta, personal communication, 2014). piloderma sphaerosporum jülich specimen examined: karelia borealis, lieksa, kitsi i, 7023:3689, 19.viii.1999 maarit similä & mari niemi 325/1999, det. mk (tur), on pinus sylvestris; karelia borealis, lieksa, säynäsemä, 7038:3683, 23.viii.1999 maarit similä & mari niemi 517/1999, det. mk (tur), on pinus sylvestris; ostrobottnia ultima, rovaniemi, pisavaara strict nature reserve, ucs 7359311:3416972, 12.ix.2013, mk 11b/13 & pekka helo (oulu), on pinus sylvestris. new to middle boreal, northern carelia – kainuu (3b) and middle boreal, southwestern lapland (3c). polyporus squamosus (huds. : fr.) fr. specimen examined: ostrobottnia ultima, rovaniemi, kukanniemi, ucs 737736:344429, 18.vi.2013, tk 899 (tur), on stumps of salix caprea at riverbank. in finland there are two different ecological forms. this belongs to the rarer one, that is found on betula and salix stumps at sea, lake and river shores. new to northern boreal, north ostrobothnia (4b). postia immitis (peck) niemelä specimen examined: regio aboënsis, kemiönsaari, dragsfjärd, kuggskär, ucs 6638:3240, 30.ix.2008, pk 3991, det. jp, (tur), on alnus glutinosa, dead standing trunk, diam. 23 cm, decay stage 1. data deficient. there are five earlier records in finland: jomala (alandia), three sites in helsinki (nylandia) and kirkkonummi (nylandia). subtrates have been fraxinus, betula, alnus incana and juglans ailanthifolia. postia perdelicata (murrill) m.j. larsen & lombard specimen examined: tavastia australis, pirkkala, kaitalankulma, saukkolampi, ucs 68192:33248, 22.viii.2013, unto söderholm 4608, det. viacheslav spirin (oulu), on pinus sylvestris. endangered. new to southwestern finland and southern ostrobothnia (2a). postia tephroleuca (fr.) jülich specimen examined: lapponia enontekiensis, enontekiö, pallas-ounastunturi national park, suastunturi e, ucs 7573150:3376710, 25.viii.2013, jp 2546 (h, j.p.), on pinus sylvestris fallen trunk, diam. 35 cm, decay 2, mixed pine dominated heath forest. new to northern boreal, forest lapland (4c). pseudotomentella nigra (höhn. & litsch.) svrček specimen examined: lapponia kittilensis, kolari, pallas-ounastunturi national park, pyhätunturi s, ucs 7510511:3383718, 9.viii.2013, jp 2415 (h, j.p.), on populus tremula fallen trunk, diam. 12 cm, decay stage 4, mixed pine dominated forest. new to northern boreal, north ostrobothnia (4b). 231© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) radulomyces confluens (fr. : fr.) m.p. christ. specimen examined: lapponia kittilensis, kolari, pallas-ounastunturi national park, kukastunturinlehto, ucs 7507610:3382076, 8.viii.2013, jp 2405, det. mk, (h, j.p.), on unknown deciduous tree, diam. 10 cm, decay stage 3, moist mixed spruce dominated forest. new to northern boreal, north ostrobothnia (4b). ramariopsis crocea (pers. : fr.) corner specimen examined: ostrobottnia ultima, rovaniemi, sinettä, ucs 7399580:3429269, 2.ix.2013, tk 1030 (tur), on ground in herb-rich picea forest. new to northern boreal, north ostrobothnia (4b). ramariopsis tenuiramosa corner specimen examined: ostrobottnia ultima, tervola, raemäki nature reserve, ucs 734369:341733, 2.ix.2012, tk 579 (tur), on ground in calciferous herb-rich forest with dryopteris sp. new to middle boreal, southwestern lapland (3c). repetobasidium vile (bourdot & galzin) j. erikss. specimen examined: regio aboënsis, parainen, korppoo, västra tvigölpan, ucs 6674:3198, 21.vii.2010, pk 6122a, det. mk, conf. hk (tur), on alnus glutinosa, fallen broken trunk, diam. 17 cm, decay stage 4. this is the third record in finland. earlier records from tammisaari (regio aboënsis) and jyväskylä (tavastia borealis) [1]. resinicium pinicola (j. erikss.) j. erikss. & hjortstam specimen examined: regio aboënsis, parainen, nauvo, fårö, ucs 665:320, 25.–27. vii.2010, pk 6249, det. mk, conf. hk (tur), pinus sylvestris, fallen whole trunk, diam. 20 cm, decay stage 1; regio aboënsis, kemiönsaari, örö, ucs 68428:32380, 28.ix.2013, coll. & det. unto söderholm 4622, conf. hk (tur/h), on fallen trunk of pinus silvestris. there is only one earlier record in finland: parainen (regio aboënsis) in 2000–2001 and fruitbody grew there on pinus sylvestris [1]. sidera vulgaris (fr.) miettinen specimen examined: nylandia, kirkkonummi, kuokkamaa nature reserve, ucs 6670640:3367597, 28.x.2009, keijo savola s281009, det. juha kinnunen, on picea abies, whole fallen trunk, diam. 7 cm, decay stage 4. this is the second record in finland. earlier it has been found only from nauvo (regio aboënsis) 1997 where it grew on picea abies [21]. sistotrema dennisii malencon specimen examined: lapponia sompiensis, pelkosenniemi, pyhä-luosto national park, huttujärvi s, ucs 7435172:3500868, 10.ix.2013, jp 2669 (h, j.p.), on burned pinus sylvestris fallen trunk, diam. 30 cm decay stage 3, burned pine dominated sub-xeric heath forest. data deficient. new to northern boreal, north ostrobothnia (4b). 232© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) tomentella coerulea (bres.) höhn. & litsch. specimen examined: lapponia enontekiensis, enontekiö, pallas-ounastunturi national park, hetta, onnasjärvet, ucs 7588466:3366002, 29.viii.2013, jp 2588 (h, j.p.), on salix caprea fallen branch, diam. 10, decay stage 3, mixed pine dominated forest. new to northern boreal, forest lapland (4c). this is the second record in finland. the first record is from pisavaara strict nature reserve in rovaniemi (ostrobottnia ultima) and it was made in 1980 [1]. tomentella stuposa (link) stalpers specimen examined: ostrobottnia ultima, rovaniemi, pisavaara strict nature reserve, sorvanulkki, ucs 7358592:3416681, 3.x.2013, mk 52/13 (oulu), on picea abies. new to middle boreal, southwestern lapland (3c). trechispora kavinioides de vries specimens examined: ostrobottnia ultima, tervola, pisavaara strict nature reserve, liljalaki, ucs 7353795:3414058, 19.ix.2013, mk 37/13 & anna-liisa ruotsalainen (oulu), on picea abies; lapponia kittilensis, kolari, pallas-ounastunturi national park, pyhätunturi nw, ucs 7512609:3382953, 3.ix.2013, jp 2639 (h, j.p.), on pinus sylvestris, fallen trunk., diam. 15 cm, decay stage 3, old and moist, mixed pine dominated forest. new to middle boreal, southwestern lapland (3c) and northern boreal, north ostrobothnia (4b). trechispora laevis k.h. larss. specimen examined: ostrobottnia ultima, rovaniemi, pisavaara strict nature reserve, sorvanulkki, ucs 7358390:3416346, 3.x.2013, mk 49/13 (oulu), on coniferous tree. new to middle boreal, southwestern lapland (3c). there are seven earlier records from finland, from the hemiboreal or south boreal zones parainen (3 records), helsinki, lammi, padasjoki and suomussalmi [1,2,5]. xylodon pruni (lasch) hjortstam & ryvarden specimen examined: regio aboënsis parainen, korppoo, wattkast, nystu, ucs 6686910:3202985, 10.x.2013, pk 8206, det. mk, conf. hk (tur), on juniperus communis, stump diam. 18 cm, decay stage 2. there are three earlier records in finland: finström (alandia), karjalohja and tammisaari (regio aboënsis). fruitbodies have been found on decorticated ulmus glabra and twig of thuja plicata [1]. discussion the new records of aphyllophoroid fungi presented in this paper are mainly from expected regions, located near to the earlier known area of their distribution. however, the records of some species are far away from the earlier findings: clavulina rugosa and trechispora laevis were found from lapland in northern finland but earlier they were known only from southern finland. this means that the distance between the new and the old localities of these fungi is hundreds of kilometers. the most remarkable records in this paper are odonticium septocystidia and junghuhnia fimbriatella, the second records in finland. both species are considered to be rare 233© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) worldwide, despite that these have been found in many european countries, russia and north america [15]. also ceraceomyces sulphurinus seems to be rare in reality despite the fact that its colorful basidiocarp is easy to notice. only three records have been made in finland, and occurrences exist in seven other countries in europe [15]. in addition to these rarely found species there is only one earlier record in finland for the following species: clavaria atroumbrina, hyphodontia flavipora, resinicium pinicola, sidera vulgaris and tomentella coerulea. resinicium pinicola seems to occur in maritime areas, all three records in finland are from the archipelago of the baltic sea. sidera vulgaris is a common species in estonia, so it is possible that in the future it will become more common in finland as well. it looks like that pseudotomentella nigra is a northern species in finland: all the finnish records are from lapland. the high amount of new information has accumulated after the publication of the finnish checklist of aphyllophorales [1]. this indicates that the aphyllophoroid fungi are still poorly known in finland. in general, for many aphyllophoroid fungi species the few records are from distant geographic locations and it is difficult to imagine that their distributions would be so scattered in reality. most likely there are numerous sites where many of these little collected species occur between the scattered current occurrences. to establish which species are truly rare, more effort should be put on inventories. occurrence of aphyllophorales, especially corticioids and clavarioids are poorly known in all parts of finland, especially in åland islands, ostrobothnia and lapland. the archipelago area and lapland are biogeographically inadequately known parts of finland because of their somewhat remote location. these areas are also hard to access by researchers. for example, there are 423 species of aphyllophorales found in the åland islands [1,5]. comparing this amount of species to numbers in other biogeographic provinces in southern finland, it is obvious that dozens of species can still be found there. traditionally species inventories have been mainly done in protected areas, but for example in southern finland only 2.3% of forests are strictly protected [22], and therefore very large areas are not ever inventoried. also our records are mainly from protected areas. accumulation of knowledge of other aphyllophorales than polypores is slow and quite sporadic because there are only few researchers working with aphyllophorales in finland. particularly comparing this effort to vast forest area of 22.8 million hectares in finland [22]. of course certain portion of aphyllophoroid fungi are rare in reality, for example on the basis of specialization in certain habitat or substrate. the fungal community occupying the smallest dead wood fractions seems to be especially poorly known [23]. the fruiting bodies of many corticioids are so small and inconspicuous that these are hard to find. this is one reason why accumulation of records of some genera is slow. also taxonomical problems and changes are confusing, and this is not a tempting point of view for amateur mycologists. acknowledgments we are grateful to ph.d. kaisa junninen in metsähallitus natural heritage services who gave us results of many polypore inventories made in the state owned forests, ph.d. heikki kotiranta in finnish environment centre who gave valuable help with identifications of some specimens and sanna-mari kunttu who drew a map. the following persons have made collections or identified specimens: maija-liisa heinonen, pekka heinonen, pekka helo, kaisa junninen, juha kinnunen, viktor kučera, mauri lahti, olli manninen, anni markkanen, 234© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) tuomo niemelä, mari niemi, esteri ohenoja, keijo savola, maarit similä, viacheslav spirin, unto söderholm and terho taarna. pk is grateful to nordenskiöld-samfundet i finland r.f. for financial support. authors’ contributions the following declarations about authors’ contributions to the research have been made: gave an original idea, wrote the first version of the manuscript and finished it, made collections: pk; made collections and commented the manuscript: jp; tk; made collections and identifications of specimens and commented the manuscript: mk; made collections (here noted with her girl’s name mari e. niemi) and commented, corrected and finished the manuscript: ms. references 1. kotiranta h, saarenoksa r, kytövuori, i. aphyllophoroid fungi of finland. a check-list with ecology, distribution, and threat categories. norrlinia. 2009;19:1–223. 2. kunttu p, kosonen t, kulju m, kotiranta h. phlebia cremeoalutacea new to finland and new records of rare corticioid fungi (basidiomycota). karstenia. 2009;49:69–71. 3. kunttu p, kulju m, kotiranta h. rare corticioid fungi in finland – records of new and little collected species (basidiomycota). karstenia. 2010;50:35–44. 4. kunttu p, kulju m, pennanen j, kotiranta h, halme p. additions to the finnish aphyllophoroid fungi. folia cryptogam est. 2011;48:25–30. 5. kunttu p, kulju m, kotiranta h. new national and regional biological records for finland. 2. contributions to the finnish aphyllophoroid funga (basidiomycota). memoranda soc fauna flora fennica. 2012;88:61–66. 6. kunttu p, pennanen j, helo t, kulju m, söderholm u. new national and regional biological records for finland. 4. additions to the knowledge of finnish aphyllophoroid funga (basidiomycota). memoranda soc fauna flora fennica. 2013;89:119-124. 7. kotiranta h, larsson kh. sistotrema luteoviride sp. nov. (cantharellales, basidiomycota) from finland. acta mycol. 2013;48:219–222. http://dx.doi.org/10.5586/am.2013.023 8. kotiranta h, shiryaev a. notes on aphyllophoroid fungi (basidiomycota) in kevo, collected in 2009. kevo notes. 2013;14:1–22. 9. spirin v, miettinen o, pennanen j, kotiranta h, niemelä t. antrodia hyalina, a new polypore from russia, and a. leucaena, new to europe. mycol prog. 2013;12:53‒61. http://dx.doi.org/10.1007/s11557-012-0815-0 10. heikinheimo o, raatikainen m. grid references and names of localities in the recording of biological finds in finland. notulae entomologicae. 1981;61:133–154. 11. knudsen h, vesterholt j, editors. funga nordica. agaricoid, boletoid and cyphelloid genera. copenhagen: nordsvamp; 2008. 12. ahti t, hämet-ahti l, jalas j. vegetation zones and their sections in northwestern europe. ann bot fenn. 1968;5:169–211. 13. hämet-ahti l. the boreal zone and its biotic subdivision. fennia. 1981;159(1):69–75. 14. miettinen o, larsson kh. sidera, a new genus in hymenochaetales with poroid and hydnoid species. mycol prog. 2011;10:131–141. http://dx.doi.org/10.1007/s11557-010-0682-5 15. bernicchia a, gorjón sp. corticiaceae s.l. italy: candusso edizioni; 2010. (fungi europaei; vol 12). 16. hjortstam k, ryvarden l. a checklist of names in hyphodontia sensu stricto – sensu lato and schizopora with new combinations in lagarobasidium, lyomyces, kneiffiella, schizopora and xylodon. synopsis fungorum. 2009;26:33–55. 17. kotiranta h, junninen k, saarenoksa r, kinnunen j, kytövuori i. aphyllophorales & heterobasidiomycetes. in: rassi, p, hyvärinen, e, juslén, a, mannerkoski, i, editors. the 2010 red list of finnish species. helsinki: ympäristöministeriö & suomen ympäristökeskus; 2010. p. 249–263. 18. renvall p. community structure and dynamics of wood-rotting basidiomycetes on decomposing conifer trunks in northern finland. karstenia.1995;35:1–51. 19. niemelä t. guide to the polypores of finland. botanical bulletins of the university of helsinki. 2011;192: 1–114. http://dx.doi.org/10.5586/am.2013.023 http://dx.doi.org/10.1007/s11557-012-0815-0 http://dx.doi.org/10.1007/s11557-010-0682-5 235© the author(s) 2014 published by polish botanical society acta mycol 49(2):221–235 kunttu et al. / noteworthy records of aphyllophoroid fungi in finland (basidiomycota) 20. eriksson j, hjortstam k, ryvarden l. mycoaciella – phanerochaete. oslo: fungiflora; 1978. (the corticiaceae of north europe; vol 5). 21. vauras j. macrofungi of the southwestern archipelago national park. metsähallituksen luonnonsuojelujulkaisuja a. 2000;112:1–91. 22. finnish forest research institute. finnish statistical yearbook of forestry. sastamala: finnish forest research institute; 2012. 23. juutilainen k, halme p, kotiranta h, mönkkönen m. size matters in studies of dead wood and woodinhabiting fungi. fungal ecol. 2011:4:342–349. abstract introduction material and methods results list of species discussion acknowledgments authors’ contributions references 2014-12-31t17:31:08+0000 polish botanical society 2014-09-05t16:22:55+0200 polish botanical society 2014-09-05t16:23:37+0200 polish botanical society 2014-11-20t23:12:52+0000 polish botanical society 2014-11-20t23:12:55+0000 polish botanical society 2014-11-20t23:12:10+0000 polish botanical society 2014-11-20t23:15:12+0000 polish botanical society 2014-09-05t16:24:42+0200 polish botanical society 2014-11-20t23:11:32+0000 polish botanical society 2014-11-20t23:12:26+0000 polish botanical society 2014-11-20t23:13:41+0000 polish botanical society 2014-09-05t16:24:18+0200 polish botanical society 2014-11-20t23:15:51+0000 polish botanical society characteristics and diversity of rhizoctonia spp. population in soil of selected forest bare-root nurseries in poland 279this is an open access article distributed under the terms of the creative commons attribution 3.0 license (creativecommons.org/licenses/by/3.0/), which permits redistribution, commercial and non-commercial, provided that the article is properly cited. © the author(s) 2014 published by polish botanical society acta mycologica introduction coniferous species cover 69.9% of the total forest area in poland, with scots pine growing on 59.5% of the forest area of all forms of ownership [1]. the large percentage of pine stands in polish forests requires constant renewal and therefore continuous production of large quantities of high-quality planting stock for reforestation is necessary. scots pine seedlings, both in nurseries and in natural stands, are vulnerable to attack by various pathogens [2–4].the most prevalent and severe disease of conifer seedlings is damping-off, the more frequently occurring, the longer a nursery is being operated, which is associated with accumulation of inoculum in soil [3,4]. original research paper acta mycol 49(2):279–290 doi: 10.5586/am.2014.025 received: 2014-10-05 accepted: 2014-12-21 published electronically: 2014-12-31 characteristics and diversity of rhizoctonia spp. population in soil of selected forest bare-root nurseries in poland marta bełka*, małgorzata mańka department of forest pathology, faculty of forestry, poznań university of life sciences, wojska polskiego 71c, 60-625 poznań, poland abstract fourty three rhizoctonia isolates obtained from four forest nurseries situated in the wielkopolska region (central-western poland) has been proved as multinucleate (anamorph – r. solani). they represented four anastomosis groups (ag): ag1-ic, ag-5, ag4-hg2 and ag2-1. three ags were found in jarocin nursery (ag-5, ag4-hg2 and ag2-1), two in łopuchówko (ag-5 and ag4-hg2) and one in konstantynowo (ag1-ic) and pniewy (ag-5). all isolates were highly pathogenic to scots pine (pinus sylvestris) seedlings and pose a large damping-off threat to the seedlings in the nurseries with single ag and in those where more ags exists. keywords: rhizoctonia spp.; anastomosis group; pathogenicity; forest nursery; pinus sylvestris; damping-off * corresponding author. email: marticz@poczta.onet.pl handling editor: maria rudawska http://creativecommons.org/licenses/by/3.0/ mailto:marticz%40poczta.onet.pl?subject=am.2014.025 280© the author(s) 2014 published by polish botanical society acta mycol 49(2):279–290 bełka and mańka / rhizoctonia spp. in forest nurseries the most severe pathogens causing damping-off in polish forest nurseries include species from genera: fusarium, rhizoctonia, pythium, alternaria and cylindrocarpon [3,5–8]. fungi belonging to the rhizoctonia genus represent a wide range of pathogenic, non-pathogenic and mutualistic species [9]. most studies on this group of fungi, known to attack more than 200 different species of plants, relate to agricultural plants, while only few researchers have studied this group of organisms in forest nurseries [4,6,10–22]. fungi classified as rhizoctonia spp. are a collection of fungi, in which the differences relate to the stage of anamorph, teleomorph, size and shape of sclerotia, and color of the mycelium [23–30]. the first reports on anastomosis reactions between isolates of r. solani date back to 1920 and 1930 [31,32]. however, only since the emergence of the anastomosis groups (ags) concept within rhizoctonia spp. [33], it has become an important tool in understanding the genetic diversity of this complex of fungi, considering the fact that isolates subscribed to different anastomosis groups can differ in pathogenicity to their host plants [34]. although rhizoctonia spp. are an important problem in a number of forest nurseries in poland but until recently, there has been little research in this area. the fact that hyphal anastomosis groups in rhizoctonia spp. are not always host specific is well known and most isolates from different anastomosis groups are capable of causing infection on a variety of plant species [24]. because virulence and host range of ags is differentiated, therefore knowledge about affinity of isolates of r. solani to particular anastomosis group is very important, in view of protection of scots pine seedlings not only in wielkopolska region but in the entire country. the aim of this study was to investigate the interand intra-species diversity within rhizoctonia spp. isolated from soil of four selected forest nurseries in the poznań region (central-western poland) in connection with their pathogenicity diversity and the resulting threat to scots pine seedlings in polish nurseries. material and methods collecting isolates soil samples were harvested from the top 15 cm of the soil profile from nurseries of forest districts: jarocin, łopuchówko, konstantynowo and pniewy. the soil was collected in early spring, before any chemicals were used. samples were placed in sterile cotton bags and stored at 5°c for no longer than two days. rhizoctonia isolates were obtained from soil samples using two trapping methods. the first method based on the use of pine seeds which were sown (25 seeds per pot) to a volume of 700 ml of unified soil from a nursery. then, after emergence of the first seedlings observation was carried out in order to capture the very first symptoms of damping-off. the symptomatic seedlings were disinfected with 0.5% sodium hypochlorite for 5 minutes, 70% ethanol (1 minute) and washed three times in distilled sterilized water, for five minutes jointly. next, the symptomatic parts of seedlings (up to 2 cm) were transferred onto martin-johnson agar medium, supplemented with antibiotics (streptomycin 0.05 g/l and aureomycin 0.05 g/l) to prevent the growth of bacteria. all isolates growing out of them were transferred onto potato dextrose agar (pda medium). 281© the author(s) 2014 published by polish botanical society acta mycol 49(2):279–290 bełka and mańka / rhizoctonia spp. in forest nurseries the second method used in the study was a modified method described by paulitz and schroeder [35] where wooden toothpicks were used. toothpics (10 per 700 ml of soil in pot) were inserted into the soil to a depth of 5 cm, evenly spaced in the pot. after 48 hours, toothpicks were removed and placed on petri plates with pda, supplemented with antibiotics (as described above). the plates with two toothpics per plate were incubated for 24 hours at room temperature. all mycelia growing from tothpicks were transferred onto pda plates in order to identify rhizoctonia isolates. rhizoctonia identification microscopic examination by cell nuclei staining. determining the number of nuclei in the cells of the rhizoctonia spp. is an important part of the process of assigning isolates to this genus [28]. the number of nuclei per cell was determined with the method described by bandoni [36]. the average number of nuclei was calculated for 50 cells of each isolate. molecular identification. dna isolation. in order to isolate dna, isolates were grown on liquid broth for 6–8 days. after that time the mycelium was placed on sterile filter paper and the excessive medium was removed. the dried mycelium was put into eppendorf tubes (1.2 ml), and placed for 24 hours at −20°c. the next step was freeze-drying of the frozen mycelia in a lyophilizer (christ alpha 1-2 ld) at −55°c and a pressure of 10−2 bar. after at least 18 hours, the mycelium was crushed with metal rods, then the cap was secured with parafilm, and the tubes were stored at −20°c until the dna extraction. dna extraction was carried out using a dneasy plant mini kit (qiagen) according to manufacturer’s recommendations. pcr-rflp analysis of rdna-its region. isolates obtained in the study have been first separated based on morphological differences and the number of nuclei per cell. in order to assign isolates to the anastomosis subgroups the restriction enzymes digestion of rflp of rdna-its regions was performed. primers its4 (tcctccgcttattgatatgc) and its5 (ggaagtaaaagtcgtaacaagg) were used for amplification of the nuclear rdna-its region [37]. the pcr amplification reactions were conducted by adding 2 μl genomic dna to 23 μl of pcr-mix, containing 2.5 μl 10× pcr buffer (qiagen), 5 μl q-solution (qiagen), 0.5 μl dntps (10 mm, fermentas gmbh), 1.75 μl of each primer (10 μm), 0.15 μl taq polymerase (fermentas gmbh) and 11.35 μl sterile highly purified water. the denaturation step at 94°c for 10 min was followed by 35 cycles of 1 min at 94°c, 1 min at 60°c and 1 min at 72°c. cycling ended with a final extension step at 72°c for 10 min (biometra, t-gradient thermoblock). after the dna extraction and restriction enzymes digestion of rflp of rdna-its regions the results of electrophoresis were checked under the uv light, and the resulting patterns were compared with those presented in the work of guillemaut et al. [38]. four restriction enzymes (msei, avaii, hincii, and muni) were used in the study. sequence analysis of its-rdna region. sequencing of its-rdna region and previous cleaning of the dna has been carried out in the laboratory of medical genetics, cb dna (mickiewicza 31, 60-835 poznań). the sequencing results were then used for further work. the sequences of rdna-its region were compared with sequences available in the genbank (http:// www.ncbi.nlm.nih.gov/genbank) to check and compare if the results of those carried with guillemault et al. [38] method were the same. characteristics of rhizoctonia isolates. growth rate measurement. fungal colonies were grown at 21°c, according to the work of tewoldemedhin et al. [39]. http://www.ncbi.nlm.nih.gov/genbank http://www.ncbi.nlm.nih.gov/genbank 282© the author(s) 2014 published by polish botanical society acta mycol 49(2):279–290 bełka and mańka / rhizoctonia spp. in forest nurseries the mycelial disc (0.5 cm diameter) of each isolate (4 reps) was placed in the center of petri dishes with pda. the petri dishes were incubated in the dark. the growth rate of individual isolates on pda was measured every 24 hours, in two directions on each plate. pathogenicity of rhizoctonia isolates. plastic pots (500 ml volume) were filled up to 2/3 with sterilized soil (120°c for 2 hours, for three days in a row). on the soil surface pda medium overgrown with seven-day-old mycelium removed from a petri dish was placed and covered with a further layer of soil (about 1 cm) into which the scots pine seeds were sown (25 seeds per pot). seeds were previously surface sterilized for 10 minutes in 30% h2o2, then rinsed with distilled water three times. thus prepared pots, and the control pots (with sterile pda medium) were watered with tap water as needed. after the first seedlings were observed emerging from the soil, the pots were checked daily and infested plants were counted. for each isolate four reps were carried out. results collected isolates of all the soil samples collected from the surveyed nurseries 43 isolates (tab. 1) classified as rhizoctonia were obtained, 24 isolates from forest nursery belonging to jarocin forest district, 8 from łopuchówko, 7 from pniewy and 4 from konstantynowo. all the isolates were multinucleate, that is representing r. solani anamorph. no binucleate or uninucleate rhizoctonia were found in the surveyed nurseries. the lowest average number of nuclei was observed in ag1-ic isolates obtained from konstantynowo (tab. 2). the highest number of nuclei per cell was observed in ag-5 isolates obtained from jarocin, with the average number of 9.5 nuclei per cell (ag-5 isolates from łopuchówko and pniewy average 8.853 and 8.637 nuclei per cell, accordingly). rhizoctonia ag2-1 was characterized by the average number of 6.02 nuclei per cell while ag4-hg2 by 5.5 and 7.268 nuclei per cell (jarocin and łopuchówko, accordingly). characteristics of rhizoctonia isolates mycelium growth rate. on average, isolates belonging to ag-5 were characterized by the fastest growth within all isolates (13.9 mm/24 h), and the slowest growing were those assigned to ag4-hg2 (8.91 mm/day). the greatest differences in growth were observed within ag-5, and the least within ag4-hg2 (fig. 1). pathogenicity of rhizoctonia isolates. differentiation within anastomosis groups in terms of pathogenicity was statistically significant for all nurseries (at a confidence level of 0.001; fig. 2). all isolates, regardless of the place of origin of the individual anastomosis groups, were characterized by high pathogenicity to pine seedlings. the most pathogenic isolates proved those belonging to ag-5 isolated from łopuchówko (100% mortality). the least pathogenic ones were found to be ag4-hgii isolates, wherein the average pathogenicity of all the isolates tested was 92.81% (fig. 2). anastomosis groups. rhizoctonia isolates obtained in the study have been first separated based on morphological differences and the number of nuclei per cell. after the dna extraction and restriction enzymes digestion of rflp of rdna-its regions the results of electrophoresis were checked under the uv light, and the resulting patterns were compared with those presented in the work of guillemaut et al. [38] (fig. 2). 283© the author(s) 2014 published by polish botanical society acta mycol 49(2):279–290 bełka and mańka / rhizoctonia spp. in forest nurseries no. isolate anamorph teleomorph anastomosis group/subset accession no. percentage sequence simillarity konstantynowo 1 140712 r. solani t. cucumeris ag1-ic fj746941.1 99 2 140711 r. solani t. cucumeris ag1-ic fj746941.1 99 3 140707 r. solani t. cucumeris ag1-ic fj746941.1 99 4 140729 r. solani t. cucumeris ag1-ic fj746941.1 99 jarocin 5 130750 r. solani t. cucumeris ag-5 gu055587.1 99 6 130711 r. solani t. cucumeris ag-5 gu055587.1 99 7 130724 r. solani t. practicola ag4-hg2 ab000032.1 99 8 130725 r. solani t. practicola ag4-hg2 ab000032.1 99 9 130728 r. solani t. practicola ag4-hg2 ab000032.1 99 10 130714 r. solani t. practicola ag4-hg2 ab000032.1 99 11 130704 r. solani t. practicola ag4-hg2 ab000034.1 99 12 130703 r. solani t. practicola ag4-hg2 ab000034.1 99 13 130733 r. solani t. cucumeris ag2-1 ay154317.1 99 14 130710 r. solani t. cucumeris ag2-1 ay154317.1 99 15 130734 r. solani t. cucumeris ag2-1 ay154317.1 99 16 130712 r. solani t. cucumeris ag2-1 ay154317.1 99 17 130707 r. solani t. cucumeris ag2-1 fj492102.3 99 18 130745 r. solani t. cucumeris ag2-1 ay154317.1 99 19 130752 r. solani t. cucumeris ag2-1 fj492102.3 99 20 130718 r. solani t. cucumeris ag2-1 fj492102.3 99 21 130744 r. solani t. cucumeris ag2-1 ab054853.1 99 22 130713 r. solani t. cucumeris ag2-1 ay154317.1 99 23 130723 r. solani t. cucumeris ag2-1 ay154317.1 99 24 130729 r. solani t. cucumeris ag2-1 ay154317.1 99 25 130772 r. solani t. cucumeris ag2-1 ay154317.1 99 26 130764 r. solani t. cucumeris ag2-1 ay154317.1 99 27 130765 r. solani t. cucumeris ag2-1 fj492102.3 99 28 130794 r. solani t. cucumeris ag2-1 fj492102.3 99 łopuchówko 29 150711 r. solani t. practicola ag4-hg2 ab000020.1 99 30 150708 r. solani t. practicola ag4-hg2 ab000020.1 99 31 150703 r. solani t. practicola ag4-hg2 hq629864.1 99 32 150710 r. solani t. practicola ag4-hg2 ab000020.1 99 33 150712 r. solani t. practicola ag4-hg2 hq629864.1 99 34 150702 r. solani t. cucumeris ag-5 eu244843.1 99 35 150707 r. solani t. cucumeris ag-5 eu244843.1 99 36 150709 r. solani t. cucumeris ag-5 eu244843.1 99 tab. 1 anastomosis groups and species of rhizoctonia isolates studied. 284© the author(s) 2014 published by polish botanical society acta mycol 49(2):279–290 bełka and mańka / rhizoctonia spp. in forest nurseries on this basis, the isolates were assigned to particular anastomosis groups. it has also been verified by comparing the its-rdna sequences of all isolates with sequences deposited in genbank (tab. 1). isolates assigned to ag1-ic have only been found in konstantynowo. they were characterized by the largest similarity to the isolates deposited in the genbank database under fj746941.1 accession number. the isolate deposited in genbank has been isolated from zoysiagrass (zoysia sp.) where it has been a causal agent of large patch disease. jarocin’s isolates assigned to ag-5 showed a high homology (99%) with isolate gu055587.1, obtained from agricultural soil from lower austria. ag-5 isolate obtained from łopuchówko was similar in 99% to those obtained from switzerland (eu244843.1), whereas those obtained from pniewy showed high homology with isolates hq898767.1, associated with potato tubers in france, and isolate hq629863.1 affecting pisum sativum in north dakota. no. isolate anamorph teleomorph anastomosis group/subset accession no. percentage sequence simillarity pniewy 37 160710 r. solani t. cucumeris ag-5 hq898767.1 99 38 160702 r. solani t. cucumeris ag-5 hq629863.1 99 39 160704 r. solani t. cucumeris ag-5 hq629863.1 99 40 160712 r. solani t. cucumeris ag-5 hq898767.1 99 41 160703 r. solani t. cucumeris ag-5 hq898767.1 99 42 160729 r. solani t. cucumeris ag-5 hq898767.1 99 43 160705 r. solani t. cucumeris ag-5 hq898767.1 99 tab. 1 (continued) anastomosis group jarocin konstantynowo łopuchówko pniewy mean s.d. mean s.d. mean s.d. mean s.d. ag-5 9.50a 0.707     8.853a 1.263 8.637 0.989 ag1-ic 4.700 0.294 ag2-1 6.02b 0.830 ag4-hg2 5.50b 0.775 7.268a 1.345 nir0.05 1.69 2.36 p > f <0.001       0.151       tab. 2 the average number of nuclei per cell in different anastomosis groups (mean value and standard deviation – s.d.; univariate analysis followed by nir test was carried out only for two nurseries). 285© the author(s) 2014 published by polish botanical society acta mycol 49(2):279–290 bełka and mańka / rhizoctonia spp. in forest nurseries fig. 1 growth rate of different anastomosis groups of rhizoctonia spp. used in the study (boxand-whisker plot: upper whisk – highest value; upper quartile; median; lower quartile; lower whisk – lowest value). fig. 2 pathogenicity of isolates of rhizoctonia spp. belonging to different anastomosis groups (box-and-whisker plot: upper whisk – highest value; upper quartile; median; lower quartile; lower whisk – lowest value). 286© the author(s) 2014 published by polish botanical society acta mycol 49(2):279–290 bełka and mańka / rhizoctonia spp. in forest nurseries isolates described in this work as ag4-hg2 were found in two nurseries – jarocin and łopuchówko. from among ag4-hg2 isolates from jarocin four were highly similar to ab000032.1, whereas two were more similar to ab000034.1. both sequences deposited in genbank were obtained from inner mongolia. isolates from łopuchówko were most similar to hq629864.1 isolated from pisum sativum in north dakota (isolates 150703 and 150712) and ab000020.1 isolated from potato in inner mongolia (isolates 150708, 150710, 150711). the biggest number of isolates (16) obtained in this study belonged to ag2-1. all of them were isolated from jarocin. they were highly homological with ay154317.1, fj492102.3 and ab054853.1 deposited in genbank. unfortunately, in the genbank database there is no information from what plant isolate ay154317.1 has been obtained. the isolate fj492102,3 was isolated from sugar beet in the usa. isolate deposited in the genbank under the number ab054853.1 has been obtained in italy from nicotiana tabacum. discussion in this study, 43 rhizoctonia isolates were obtained from four forest nurseries and their anastomosis groups (ags) and anastomosis subgroups were determined. they all proved to be r. solani, a species widely distributed in polish forest nurseries. our understanding concerning the occurrence of rhizoctonia anastomosis groups in polish forest nurseries is very scarce. knowledge of rhizoctonia spp. ags occurring in the nursery could facilitate the selection of plant protection methods and products to protect the seedlings against fig. 3 example of rflp analysis of the its rdna region for isolates of 150707, 150709 and 160705 (ag-5) of rhizoctonia solani amplified with primers its4 and its5 and digested with muni. 287© the author(s) 2014 published by polish botanical society acta mycol 49(2):279–290 bełka and mańka / rhizoctonia spp. in forest nurseries these pathogens. the results of some studies [40–42] point to different sensitivity to fungicides of isolates belonging to different ags. on the other hand, the works of herr [43] and sharon et al. [44] suggest the possibility of using non-pathogenic strains as biological control agents against highly pathogenic strains of rhizoctonia. pathogenicity tests of different ags to scots pine and norway spruce (picea abies) carried out on uninucleate isolates confirm that they are the causing agents of dampingoff in many finnish nurseries [45–47]. at the same time from norwegian and finnish forest nurseries uninucleate and binucleate strains of rhizoctonia have been isolated from healthy seedlings of scots pine and norway spruce [48–51]. so far, no uninucleate isolates were obtained from polish forest nurseries. in one of the few works on rhizoctonia spp. in forest nurseries camporota and perrin [52] have demonstrated the predominant role of r. solani as the primary pathogen causing damping-off of pine seedlings. this is also confirmed by our study where all the obtained isolates proved to be multinucleate. so far, 14 anastomosis groups (ag-1 to ag-13 and ag-b1) have been described within multinucleate rhizoctonia [53]. eight groups (ag-1 to ag-6, ag-8 and ag-9) were further subdivided into subgroups. from among those stępniewska-jarosz et al. [6] found five different anastomosis groups in polish forest nurseries. the most frequent was ag-5 (37% of isolates), followed by ag 2-1 (30%) and 27% of the isolates were identified as ag-4. groups ag 1-ib and ag 2-2 were only represented by single isolates. in our study one of the nurseries (forest nursery jarocin) examined earlier by stępniewska-jarosz et al. [6], was surveyed for rhizoctonia spp. for the second time and no binucleate isolates were found. two ags found in the previous study were also isolated: ag-5 (2 isolates) and ag 2-1 (16 isolates), while the ag4-hg2 (6 isolates) has not been previously observed. the difference may result from possible bridging of some isolates belonging to certain ags with isolates belonging to the other [24] in the meantime and/or from the fact that different techniques were used to assign isolates to certain ags. while stępniewska-jarosz et al. [6] used classic techniques, with culture of isolates in vitro and subsequent observation of their growth under microscope, our study was fully based on molecular methods. though the anastomosis groups differed significantly from each other in terms of pathogenicity, all the isolates studied proved to be highly pathogenic to scots pine seedlings. this means that the ags in question may contribute to considerable damping-off threat to the seedlings in all the nurseries. as most of the research on rhizoctonia in poland has been conducted on agricultural plants, it is highly desirable to further screen forest nurseries for the pathogen as well, both for better recognition of the situation and for the hope of finding non-pathogenic or mutualistic species, which could possibly contribute to decreasing the infection threat. acknowledgments the authors thank two anonymous reviewers for useful comments improving the manuscript. this work was due to grant financed by the ministry of science and higher education no. n n309 134335. authors’ contributions the following declarations about authors’ contributions to the research have been made: collected material, laboratory works, photographs, statistics, bibliography, draft of the manuscript: mb; 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http://dx.doi.org/10.1016/s0929-1393(98)00040-7 53. carling de, baird re, gitaitis rd, brainard ka, kuninaga s. characterization of ag-13, a newly reported anastomosis group of rhizoctonia solani. phytopathology. 2002;92:893–899. http://dx.doi.org/10.1094/ phyto.2002.92.8.893 http://dx.doi.org/10.1094/phyto-09-10-0247 http://dx.doi.org/10.1094/phyto-09-10-0247 http://dx.doi.org/10.1046/j.0028-646x.2001.00265.x http://dx.doi.org/10.1046/j.0028-646x.2001.00265.x http://dx.doi.org/10.1046/j.1439-0329.2002.00292.x http://dx.doi.org/10.1046/j.1439-0329.2002.00292.x http://dx.doi.org/10.1080/02827580601019662 http://dx.doi.org/10.1080/02827589209382746 http://dx.doi.org/10.1111/j.1439-0329.1994.tb00984.x http://dx.doi.org/10.1111/j.1439-0329.1994.tb00984.x http://dx.doi.org/10.1016/s0953-7562(09)80431-0 http://dx.doi.org/10.1016/s0953-7562(09)80431-0 http://dx.doi.org/10.1111/j.1439-0329.1995.tb00328.x http://dx.doi.org/10.1111/j.1439-0329.1995.tb00328.x http://dx.doi.org/10.1016/s0929-1393(98)00040-7 http://dx.doi.org/10.1094/phyto.2002.92.8.893 http://dx.doi.org/10.1094/phyto.2002.92.8.893 abstract introduction material and methods collecting isolates rhizoctonia identification results collected isolates characteristics of rhizoctonia isolates discussion acknowledgments authors’ contributions references 2014-12-31t17:44:22+0000 polish botanical society 2014-11-20t23:10:18+0000 polish botanical society 2014-11-20t23:10:32+0000 polish botanical society 2014-11-20t23:10:06+0000 polish botanical society 2014-09-06t20:22:35+0200 polish botanical society 2014-11-20t23:16:26+0000 polish botanical society 2014-11-20t23:11:49+0000 polish botanical society 2014-11-20t23:14:55+0000 polish botanical society 2014-11-20t23:17:39+0000 polish botanical society 2014-09-06t20:22:42+0200 polish botanical society 2014-11-20t23:13:24+0000 polish botanical society 2014-09-05t16:24:28+0200 polish botanical society 2014-11-20t23:18:21+0000 polish botanical society 2014-09-06t20:23:13+0200 polish botanical society 2014-11-20t23:16:46+0000 polish botanical society 2014-11-20t23:19:50+0000 polish botanical society 2014-11-20t23:12:43+0000 polish botanical society 2014-11-20t23:14:16+0000 polish botanical society 2014-11-20t23:12:58+0000 polish botanical society the genus fusicladium (hyphomycetes) in poland małgorzata ruszkiewicz michalska and ewa połeć 1 department of algology and mycology, university of łódź, banacha 12/16, pl 90 237 łódź mrusz@biol.uni.lodz.pl; ewa_polec@op.pl r u s z k i e w i c z m i c h a l s k a m., p o ł e ć e . : the genus fusicladium (hyphomycetes) in poland. acta mycol. 41 (2): 285 298, 2006. the paper presents new and historical data on the genus fusicladium verified on the base of the recently published critical monograph. fifteen species recorded in poland under the name fusicladium and synonymous pollaccia and spilocaea are reported; 5 are documented by authors’ materials from central poland while the other taxa are supported with literature data only, including three species belonging currently to fusicladiella and passalora. three species, reported here for the first time in poland: fusicladium convolvularum ondřej, f. scribnerianum (cavara) m. b. ellis and f. virgaureae ondřej, are known from a few localities in the world. all the species are provided with the distribution maps and the newly reported ones are illustrated with ink drawings. key words: parasitic fungi, anamorphic fungi, deuteromycotina, distribution, poland introduction worldwide 57 fungal taxa belong to the anamorphic genus fusicladium bonord. em. schubert, ritschel et u. braun. they are phytopathologically relevant pathogens, causing leaf spots, necroses, scab diseases as well as leaf and fruit deformations of members of at least 52 angiospermous plant genera (s c h u b e r t , r i t s c h e l , b r a u n 2003). the fungi are host specific, mostly confined to a single host genus or allied host genera in a single family, e.g. fusicladium pomi parasitizing the members of rosaceae. the teleomorphic stages belonging to venturiaceae (ascomycota) develop after the mycelium overwinters in plant organs. the name fusicladium was traditionally used for venturia anamorphs having sympodial (denticulate) or percurrent (annellate) conidiogenous cells. three genera were later distinguished depending on the proliferation mode: fusicladium with sympodial proliferation, pollaccia with monoblastic, determinate to percurrent conidiogenous cells (with few rather inconspicuous annellations) and spilocaea with percurrent proliferation and numerous, conspicuous annellations acta mycologica vol. 41 (2): 285-298 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 286 m. ruszkiewicz michalska and e. połeć (s c h u b e r t e t a l . 2003). the division has been accepted in many literature reports (e.g. e l l i s 1976; b r a n d e n b u r g e r 1985; r e v a y 1998). due to the number of taxa with mixed types of conidiogenous cells, these features are insufficient for the generic separation and it was not maintained by the authors of the first monograph of fusicladium (s c h u b e r t et al. 2003). they reduced names pollaccia e. bald. et cif. and spilocaea fr. to synonymy with fusicladium bonord., postulating to preserve the name instead of the older spilocaea. the decision to merge these genera is supported by light and electron microscopy research and molecular data which show that features such as the type and growth of the mycelium, arrangement of conidiophores, structure of conidiogenous loci and shape, size and formation of conidia may not be used to define various genera of anamorphs belonging to venturiaceae. these features are, however, useful for distinguishing species. properties that enable the differentiation of fusicladium from other hyphomycetous genera are as follows: mycelium and stromata usually subcuticular to intraepidermal, hyphae often radiating, forming hyphal plates, conidiophores usually erumpent through the cuticle, pigmented and with equally thickened wall, conidiogenous loci inconspicuous with always unthickened wall, conidia pigmented, mostly olivaceous, solitary or catenate, ameroto phragmosporous, 0-2(-4)-septate. materials and methods available literature data were critically analysed and used to prepare a list of fusicladium species recorded in poland and maps of their distribution. phytopathological reports of economically important species were omitted for the sake of clarity and mainly ‘floristic’ data were used below. the original material consisting of plant organs affected by fusicladium species was collected in central poland: in the region of łódź city and in the wyżyna częstochowska upland. for microscopic examination, specimens were made in lactic acid and heated with an alcohol burner. nikon smz 10 a and nikon e 400 microscopes were used to identify fungal species which were determined according to s c h u b e r t et al. (2003). features of these specimens correspond to those given there. the nomenclature of host plants is given after m i r e k et al. (2002). the study material is deposited in the herbarium universitatis lodziensis (lod) in the collection of parasitic fungi labelled as pf. results according to s c h u b e r t et al. (2003) 7 taxa are known to occur in poland, namely fusicladium betulae, f. radiosum var. populi-albae, f. radiosum var. radiosum, f. saliciperdum, fusicladium sp. and common in europe – f. pomi and f. pyrorum. a thorough review of the polish mycological literature has revealed that 6 other species have also been recorded in poland under the names fusicladium, pollaccia and spilocaea. three of these species are currently classified in different genera: 2 belong to passalora and 1 is synonymized with fusicladiella melaena. the list of species occurring in poland is supplemented with 3 species new to the country: fusicladium convolvularum, f. scribnerianum and f. virgaureae. 288 m. ruszkiewicz michalska and e. połeć fusicladium convolvularum ondřej, česká mycol. 25 (3): 171 (1971), teleomorph unknown. leaf spots amphigenous, 1-3 mm wide, brown, later with greyish brown centre and brown margin. conidiophores solitary or in small groups, arising from stromatic cells, straight, or curved at the apex, subcylindrical to geniculate-sinuous, 29.4-44.1 x 4.2-4.9 μm, 0-1(-3)-septate, pale to medium brown, smooth. conidiogenous cells terminal with a single or several conidiogenous loci, proliferation sympodial, loci unthickened, not or often somewhat darkened-refractive. conidia solitary or occasionally in unbranched or branched chains, ellipsoid-ovoid, fusiform, subcylindrical, 17.1-22.3 x 4.9-5.2 μm, 0-1-septate, mostly constricted at the septa, pale olivaceous, smooth, hila 1.9-2.5 μm wide, flat, not or somewhat darkened-refractive (fig. 2). specimens examined. on convolvulus arvensis: central poland, wyżyna częstochowska upland, at ne base of the góra lipówki bliskie hill near olsztyn, roadside, 21 aug. 1998, leg. m. ruszkiewicz-michalska, lod 885; łódź, botanical garden, sect. of medicinally and industrially important plants, weed overgrowing convallaria majalis, 21 aug. 2004, leg. e. połeć, lod 2261; łódź, “sielanka” park, near pabianicka str., weed overgrowing kerria japonica, 27 june 2005, leg. m. ruszkiewicz-michalska et m. jakiel, lod 2229 (fig. 1). notes. the fungus parasitizes members of the genera calystegia and convolvulus. it has been reported on calystegia soldanella from great britain, on calystegia sepium and convolvulus arvensis from the czech republic, and on the latter host also from new zealand (fa r r et al. 2006). the species has been observed in poland for the first time. fig. 2. fusicladium convolvularum on convolvulus arvensis: a leaf with spots, b conidio phores, c conidia; scale bars a 10 mm, b c 10 μm. 290 m. ruszkiewicz michalska and e. połeć fusicladium pomi (fr.) lind, dan. fung.: 521 (1913), ≡ f. dendriticum (wallr.) fuckel, f. orbiculatum (desm.) thüm., spilocaea pomi fr.: fr., teleomorph: venturia inaequalis (cooke) g. winter. distribution. as f. dendriticum: on malus pumila, szczecin (z a l e s k i , m a d e j 1964); on malus sylvestris, czasław, rabka, prądnik czerwony and czarna wieś in kraków (n a m y s ł o w s k i 1914), puławy, włostowice (k o n o p a c k a 1924), skierniewice (z w e i g b a u m ó w n a 1925), botanical garden in kraków (wr ó b l e w s k i 1925), near czerniawa zdrój (l a u b e r t 1931), poznań, puszczykowo, węgierki, włocławek (d o m i n i k 1936); on pyrus baccata, trzcianka (d o m i n i k 1935, 1936); as f. orbiculatum: on sorbus aucuparia, zwierzyniec (z w e i g b a u m ó w n a 1925); on sorbus latifolia (d o m i n i k 1963); as s. pomi: on malus domestica, m. pumila, m. × purpurea and malus sp.: przelewice, szczecin (m a d e j 1971, 1974); on malus spp.: kazimierski landscape park (r o m a s z e w s k a s a ł a t a et al. 1991-1992); on malus sylvestris: mielnik (r o m a s z e w s k a s a ł a t a , m u ł e n k o 1983) (fig. 3). notes. this cosmopolitan, polyphagous and economically important species has so far been noted on 12 host genera of the rosaceae family, but it mainly infects representatives of malus: m. domestica, m. prunifolia and m. sylvestris (tr i e b e l 1999; s c h u b e r t et al. 2003). fusicladium pyrorum (lib.) fuckel, jahrb. nassauischen vereins naturk. 23-24: 357 ‘1869’ (1870), as “f. pyrinum”, teleomorph: venturia pyrina aderh. specimen examined. on pyrus communis, central poland, łódź, “sielanka” park, near pabianicka str., ornamental, 27 june 2005, leg. m. ruszkiewicz-michalska et m. jakiel, lod 2206. distribution. as f. pirinum: on pyrus communis, without precised locality (c h e ł c h o w s k i 1902), czarna wieś and prądnik czerwony in kraków (n a m y s ł o w s k i 1914), puławy, kazimierz (k o n o p a c k a 1924), skierniewice (z w e i g b a u m ó w n a 1925), near czerniawa zdrój (l a u b e r t 1931), włocławek (d o m i n i k 1935, 1936), szczecin (z a l e s k i , m a d e j 1964), łeba, cetniewo (m i c h a l s k i 1967), neple, gnojno in the bug river valley (d a n i l k i e w i c z 1987); as f. pyrorum: on pyrus domestica, przelewice and szczecin (m a d e j 1974) (fig. 4). notes. the fungus infects mainly pyrus communis, but it may also be found on the members of 4 other genera belonging to the pomoideae, including malus species, which are above all parasitized by f. pomi ( s c h u b e r t et al. 2003). the latter differs from f. pyrorum by the proliferation type of conidiogenous cells (percurrent vs. sympodial) and width of loci (4-5 μm vs. 1-3 μm). fusicladium radiosum (lib.) lind var. populi-albae (m. morelet) ritschel et u. braun, ann. mycol. 3: 340 (1905), ≡ pollaccia radiosa (lib.) e. bald. et cif., teleomorph: venturia tremulae aderh. specimen examined. on populus alba: central poland, chechło ii near łódź, near the railroad, 13 sept. 2000, leg. m. ruszkiewicz, lod 2501. the dimensions of conidia (1-septate, 8.6-9.1 μm wide) correspond to the range given for this variety in the fusicladium monograph (s c h u b e r t et al. 2003). distribution. for the list of polish localities of the fungus on populus alba, p. nigra and p. tremula see m u ł e n k o (1996) (fig. 4). according to the data of s c h u b e r t et al. (2003) two varieties of this species are known from poland: var. radiosum (on p. tremula) and var. populi-albae (on p. alba). the genus fusicladium 293 arising as lateral branches of brown hyphae, conidiogenous cells polyblastic, sympodial, terminal or intercalary, with numerous conidiogenous loci and conidia in unbranched chains, 5-8 μm wide (s c h u b e r t et al. 2003). v. chlorospora is known on 7 salix species from poland (fa r r et al. 2006). fusicladium virgaureae ondřej, česká mycol. 25 (3): 170 (1971), teleomorph unknown. leaf spots amphigenous, shape and size variable, irregular, yellowish, olivaceous to brown. conidiophores solitary or in small, loose fascicles, erect, straight to somewhat flexuous, subcylindrical to slightly sinuous, unbranched or rarely branched, 36.8-63.7 x 4.9-5.4 μm, septate, yellowish or olivaceous-brown to medium brown, smooth. conidiogenous cells terminal with a single or up to four loci, proliferation sympodial. conidia in unbranched chains, cylindrical to fusiform or obclavate, straight, 13.5-15.9 x 4.4-5.2 μm, 0-1-septate, yellowish or olivaceous-brown, smooth, hila neither thickened nor darkened (fig. 7). specimen examined. on solidago serotina: central poland, wyżyna częstochowska upland, złoty potok near częstochowa, manor park, roadside, 25 aug. 1998, leg. m. ruszkiewicz, lod 886 (fig. 5). notes. f. virgaureae is the only species of this genus parasitizing plants belonging to the asteraceae. it infects only the members of the genus solidago and is known from 3 countries. occurrence of the fungus has been reported on s. gigantea from austria and on s. virgaurea from the czech republic and slovakia (s c h u b e r t et al. 2003; fa r r et al. 2006). the species is new to the polish mycobiota. fig. 6. fusicladium scribnerianum on betula pendula: a leaf with spot, b conidiophores, c conidia; scale bars: a 10 mm, b c 10 μm. 294 m. ruszkiewicz michalska and e. połeć excluded species fusicladium aronici sacc., michelia 2: 171 (1880). distribution. on doronicum austriacum and d. clusii: numerous localities in the tatra mts. (s t a r m a c h o w a 1963; m u ł e n k o et al. 2004). notes. this species currently belongs to fusicladiella melaena (fuckel) s. hughes (s c h u b e r t et al. l. c.). the genus fusicladiella differs from fusicladium by forming membranaceous mycelium and stromata, having older conidiophores usually curved and conidiogenous loci thickened and darkened. fusicladium depressum (berk. et broome) roum., fungi gall. exs. 86 (1879). distribution. for the list of host species and polish localities see m u ł e n k o (1996). notes. according to s c h u b e r t et al. (2003) and c r o u s & b r a u n (2003) this name is now synonymized with passalora depressa (berk. et broome) sacc. fusicladium heterosporum höhn., ann. mycol. 3: 337 (1905). distribution. on chamaenerion angustifolium: włoszczowa (m o e s z 1926). notes. the species is currently considered to be passalora heterospora (höhn.) höhn. (s c h u b e r t et al. l.c.; c r o u s , b r a u n l.c.). fig. 7. fusicladium virgaureae on solidago serotina: a leaf with spots, b conidiophores, c conidia; scale bars a 10 mm, b c 10 μm. the genus fusicladium 295 final remarks the first polish records of taxa currently belonging to fusicladium are dated back to the beginning of 20th century (c h e ł c h o w s k i 1902). the most of the older findings come from floristic or phytopathological investigations (n a m y s ł o w s k i 1914; l a u b e r t 1921, 1931; k o n o p a c k a 1924; wr ó b l e w s k i 1925; z w e i g b a u m ó w n a 1925; d o m i n i k 1936, 1963; s t a r m a c h o w a 1964, 1966; z a l e s k i , m a d e j 1964; m i c h a l s k i 1967; m a d e j 1971, 1974). the more recent data are provided by mycocoenological studies carried out in a few polish regions of a special floristic value: białowieża national park (m u ł e n k o 1994, 1996), tatra mts. (s a ł a t a et al. 1993) and wyżyna lubelska upland (r o m a s z e w s k a s a ł a t a , m u ł e n k o 1983; d a n i l k i e w i c z 1987; r o m a s z e w s k a s a ł a t a et al. 1991-1992). the best documented is occurrence of common, economically important species – fusicladium pomi and f. pyrorum (compare figs 3, 4). the other species are known from a few localities in poland, mainly from more intensively investigated areas. many other regions of the country, are still less studied and therefore the listed localities of these species do not present their comprehensive distribution. the 3 newly recorded taxa are known from a few countries only, on different continents, e.g. fusicladium scribnerianum has been reported from europe, asia and australia. this distribution implies that actually they are not very rare or restricted to certain areas. the assumption is supported with our findings, especially that of fusicladium convolvularum, which has been repeatedly collected in central poland since its discovery on the wyżyna częstochowska upland in 1998. acknowledgement: authors are grateful to dr. uwe braun (halle/saale, germany) for providing the es sential literature. the studies were partially supported by the university of łódź (grant no 505/396 and no 505/397). references b r a n d e n b u r g e r w. 1985. parasitische pilze an gefäßpflanzen in europa. g. fischer verlag, stutt gart new york, 1248 pp. c h e ł c h o w s k i s. 1902. spostrzeżenia grzyboznawcze (observationes mycologicae polonicae). pam. fizjogr. 17: 1 38. c r o u s p. w., b r a u n u. 2003. mycosphaerella and its anamorphs: 1. names published in cercospora and passalora. cbs biodiversity series 1: 1 571. d a n i l k i e w i c z m. 1987. grzyby pasożytnicze lewobrzeżnej doliny środkowego bugu (parasitic fungi of river bug valley). acta mycol. 23 (2): 37 80. d o m i n i k t. 1935. grzyby pasożytnicze zebrane w okolicy włocławka w sierpniu 1934 roku. acta soc. bot. pol. 12 (2): 201 205. d o m i n i k t. 1936. materiały do flory grzybów mikroskopowych zachodniej polski (beiträge zur kennt nis der mikroskopischen pilzflora westpolens). spraw. kom. fizjogr. 70: 1 72. d o m i n i k t. 1963. notatki mikologiczne z lat 1945 1960. zesz. nauk wsr szczecin 10: 47 77. d u g a n f. m., s c h u b e r t k., b r a u n u. 2004. check list of cladosporium names. schlechtendalia 11: 1 103. e l l i s m. b. 1976. more dematiaceous hyphomycetes. cab international mycological institute, kew, surrey, 507 pp. f a l i ń s k i j. b., m u ł e n k o w. 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choroby grzybowe drzew i krzewów owocowych, warzyw i roślin ozdob nych w ogrodach działkowych miasta szczecina w roku 1958 (fungus diseases of fruit trees and bu shes, vegetables and ornamental plants in allotment gardnes in the city of szczecin in 1958). roczn. wsr, poznań 19: 209 232. z w e i g b a u m ó w n a z. 1925. grzyby okolic skierniewic (les champignons des environs de skierniewi ce). acta soc. bot. pol. 2: 275 301. the genus fusicladium 297 przegląd krajowych gatunków z rodzaju fusicladium s t r e s z c z e n i e lista stwierdzonych dotąd w polsce przedstawicieli rodzaju fusicladium [= pollaccia, spilocaea] obejmuje 12 gatunków, z których 3 należą obecnie do passalora i fusicladiella. dane dotyczące 7 gatunków fusicladium pochodzą wyłącznie ze źródeł publikowanych. wy kaz uzupełniono o 3 gatunki nowe dla kraju: fusicladium convolvularum, f. scribnerianum i f. virgaureae oraz nowe stanowiska 2 taksonów notowanych uprzednio. dla wszystkich gatunków sporządzono mapy rozmieszczenia w polsce, a dla gatunków nowo stwierdzonych również opisy i rysunki struktur morfologicznych. 2014-01-01t11:44:38+0100 polish botanical society 2014-09-06t20:21:36+0200 polish botanical society 2014-11-20t23:18:51+0000 polish botanical society 2014-11-20t23:13:55+0000 polish botanical society 2014-11-20t23:13:44+0000 polish botanical society 2014-11-20t23:12:13+0000 polish botanical society 2014-11-20t23:12:16+0000 polish botanical society 2014-11-20t23:15:42+0000 polish botanical society 2014-09-06t20:22:58+0200 polish botanical society 2014-11-20t23:20:15+0000 polish botanical society 2014-11-20t23:15:15+0000 polish botanical society 2014-09-06t20:23:51+0200 polish botanical society 2014-11-20t23:09:54+0000 polish botanical society 2014-11-20t23:10:23+0000 polish botanical society 2014-11-20t23:11:54+0000 polish botanical society 2014-11-20t23:14:46+0000 polish botanical society 2014-11-20t23:10:29+0000 polish botanical society interactive physiological response of potato (solanum tuberosum l.) plants to fungal colonization and potato virus y (pvy) infection 291this is an open access article distributed under the terms of the creative commons attribution 3.0 license (creativecommons.org/licenses/by/3.0/), which permits redistribution, commercial and non-commercial, provided that the article is properly cited. © the author(s) 2014 published by polish botanical society acta mycologica original research paper acta mycol 49(2):291–303 doi: 10.5586/am.2014.015 received: 2014-07-01 accepted: 2014-09-04 published electronically: 2014-11-29 interactive physiological response of potato (solanum tuberosum l.) plants to fungal colonization and potato virus y (pvy) infection dominika thiem1, adriana szmidt-jaworska2, christel baum3, katja muders4, katarzyna niedojadło5, katarzyna hrynkiewicz1* 1 department of microbiology, faculty of biology and environment protection, n. copernicus university, lwowska 1, 87-100 toruń, poland 2 chair of plant physiology and biotechnology, faculty of biology and environment protection, n. copernicus university, lwowska 1, 87-100 toruń, poland 3 soil science, university of rostock, justus-von-liebig-weg 6, d-18059 rostock, germany 4 norika – nordring kartoffelzuchtund vermehrungs gmbh, parkweg 4, d-18190 groß lüsewitz, germany 5 department of cell biology, faculty of biology and environment protection, n. copernicus university, lwowska 1, 87-100 toruń, poland abstract potato plants can be colonized by various viruses and by symbiotic, saprophytic and pathogenic fungi. however, the significance of interactions of viral infection and fungal colonization is hardly known. this work presents a model experiment in which the influence of three different types of fungal associations on the growth and physiology of the potato variety pirol was tested individually or in combination with infection by pvy. it was hypothesized that simultaneous viral and fungal infections increase the biotic stress of the host plant, but mutualistic plant-fungal associations can mask the impact of viral infection. in the present study, a symbiotic arbsucular mycorrhizal fungus, glomus intraradices, significantly stimulated the growth of plants infected with pvy. in contrast, two saprophytic trichoderma spp. strains either did not influence or even inhibited the growth of pvy-infected plants. also, inoculation of pvy-infected potato plants with a pathogenic strain of colletotrichum coccodes did not inhibit the plant growth. growth of the pvy-free potato plants was not promoted by the symbiotic fungus, whereas t. viride, t. harzianum and c. coccodes had an evident inhibitory effect. the strongest growth inhibition and highest concentration of h2o2, as an indicator of biotic stress, was observed in pvy-free potato plants inoculated with t. harzianum and c. coccodes strains. surprisingly, ultrastructural analysis of pvy-infected plant roots colonized by g. intraradices showed virus-like structures in the arbuscules. this pointed to the possibility of mycorrhizal-mediated transmission of virus particles and has to be further * corresponding author. email: hrynk@umk.pl handling editor: tomasz leski http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.2014.015 mailto:hrynk%40umk.pl?subject=am.2014.015 292© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization introduction potato virus y (pvy) represents the most important taxonomic group of viral pathogens infecting potato (solanum tuberosum l.). it can reduce the quality of the crop and may cause yield losses up to 80% [1]. several pvy strain groups are distinguished, each of them causing different symptoms, such as mosaicism, veinal necrosis, leaf spots and systemic necrosis. some strains can also evoke tuber necrosis, particularly in sensitive potato varieties [2]. high infectivity of pvy is attributed both to its high genetic variability and cultivation of susceptible plant cultivars. transmission of the virus between host plants takes place mainly by aphids, arthropods, nematodes, fungi (mostly obligate parasites), or plasmodiophorid vectors [1,3,4], although incidentally it may be also transmitted mechanically. plants grown under high infection pressure can only be protected from infection by a consequent use of insecticides [5]. meanwhile, a number of plant-associated microorganisms can reduce the negative impact of plant pathogens [6–8]. these include arbuscular fungi, symbionts of many crops, which can stimulate the absorption of water and nutrients from the soil – mainly phosphorus (p) and nitrogen (n), and increase tolerance to abiotic and resistance to biotic stress factors, e.g., pathogen infections [9]. similar properties have also been reported for strains of some saprotrophic trichoderma spp. [10]. some strains of this genus have the ability to synthesize a number of secondary metabolites, such as plant hormones and vitamins, and can increase the availability of nitrogen (n) and phosphorus (p) or other plant nutrients. as a result, they can stimulate plant growth and productivity in either the presence or absence of other microorganisms and compete with pathogenic soil microorganisms [11]. some trichoderma strains exhibit antibiosis and mycoparasitism activities, which may lead to the reduction of plant pathogen populations [6]. it has been shown that saprophytic trichoderma spp. releases elicitors during colonization of plant roots, which activate defense pathways of the host plant, thus protecting it against pathogen infection [6]. however, antiviral effects of saprophytic fungi have not been elucidated so far. apart from the fungi promoting plant growth, there is also a large group of pathogenic fungi that colonize potato plants. these fungi can synthesize enzymes and toxins that damage plant cells or interrupt the hormonal regulation of plant growth. among them, colletotrichum coccodes, the causal agent of potato black dot, produces characteristic symptoms by formation of microsclerotia that prevail in host tissues in the late growing season [12]. pathogenic microorganisms may increase the level of reactive oxygen species (ros) in plant cells, which interferes with the metabolic processes and activates plant defense responses. such a plant response to pathogen infection was observed both for fungi and viruses [13]. examined by testing with immunoassays and real transmission to uninfected plants. in conclusion, although mycorrhiza formation might decrease the impact of pvy infection on plants, a possible role of mycorrhizal fungi as virus vectors is discussed. keywords: biotic stress; potato virus y (pvy); glomus intraradices; trichoderma viride; t. harzianum; colletotrichum coccodes 293© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization in the present work, the effects of fungal colonization of pvy-infected (pvy+) and non-pvy-infected (pvy−) potato plants are tested in a model experiment. the tested fungi represent three ecological groups: symbiotic – glomus intraradices, saprotrophic – trichoderma viride and t. harzianum, and pathogenic colletotrichum coccodes. the level of h2o2 in the leaves served as an indicator of the stress level for pvy+ and pvy− plants. additionally, fungal hyphae were examined microscopically for the intracellular presence of virus particles. material and methods plant and microbial material tubers of potato cv. pirol (norika gmbh, sanitz, germany) were used in the experiments, which was selected as model plant, since it is described to be medium resistant to infection by pvy and fungal pathogens. pvy infection of each tuber as well as the absence of the virus in the control material (pvy− tubers) were confirmed by elisa. sprouting tubers were inoculated individually by four fungal strains, differing in the impact on growth and development of potato plants: symbiotic – g. intraradices in the form of mycorrhizal inoculum (inoq, gmbh, institut für pflanzenkultur, germany), saprophytic – t. viride dar5 (collection of the university of rostock, germany) and t. harzianum k116 (collection of the n. copernicus university, toruń, poland), and pathogenic – c. coccodes k116 (collection of the norika gmbh, germany). pot experiment potato tubers were placed in polyethylene pots filled with sterile mixture of soil and vermiculite (1:1, v:v, 1 kg mixture in each pot). in the trial, 15 tubers infected with pvy and 15 control tubers (non-pvy-infected) were used. the experiment was conducted in a growth chamber under a sodium lighting system [100 μmol m−2 s−1 par (photosynthetically active radiation), temp. 24°c, 16 h light and 8 h darkness]. inoculum samples of t. viride dar5, t. harzianum and c. coccodes k116 strains were prepared in petri dishes on pda medium (potato dextrose agar, difco). strains were cultivated for 7 days at 23°c. afterwards, each fungal inoculum with an adjacent small amount of agar was transferred with a sterile scalpel onto the sprouting tubers. a sample of the mycorrhizal g. intraradices fungus (spores and hyphae) was introduced in a mixture with sand and vermiculite used in the recommended by the manufacturer amount – 20 ml/pot. inoculation of plants was repeated after a period of two weeks to enhance the effect of fungal inoculation. for each plant treatment, the following inoculation variants were performed: non-inoculated control, inoculated with: g. intraradices, t. viride dar5, t. harzianum and c. coccodes (3 replicates for each variant). this pattern of inoculation was used both for pvy-infected and non-pvy-infected plants (30 plants in total). the plants were watered with sterile distilled water. after a 10-week growth period, the plants were harvested and divided into roots, stems and leaves. leaves and roots (100 mg per each treatment) were frozen in liquid nitrogen for subsequent determination of the hydrogen peroxide (h2o2) content. the rest of the plant biomass (roots, stems and leaves) was dried at 65°c until constant weight was achieved and weighed. 294© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization determination of hydrogen peroxide (h 2 o 2 ) level analysis of h2o2 level in plant tissues (leaves, roots) was performed using the peroxidetecttm kit for determination of aqueous and lipid hydroperoxides (sigma, saint louis, missouri 63103 usa), according to the manufacturer’s protocol. the level of hydrogen peroxide was expressed as nmol/g of tissue according to the following formula: nmol of peroxide / g of plant tissue = [(a560 − a560 control sample) × dilution] / [a560 (1 nmol peroxide) × volume of sample]. electron microscopy analysis small pieces of plant root tissue inoculated with g. intraradices were fixed overnight at 4°c with 2.5% glutaraldehyde in 0.1 m phosphate-buffered saline (pbs, ph 7.2). the first hour of fixation was carried out in a vacuum pump at 4°c [12]. after washing in pbs (ph 7.2), the sample was postfixed with 1% oso4 in pbs (ph 7.2) for 2 hours at 4°c. samples were washed, dehydrated in graded concentrations of ethanol up to 100% and embedded in lr gold (sigma) with 1% benzoyl peroxide as a polymerization accelerator and polymerized at 4°c for 24 h days. ultrathin longitudinal sections were cut using a leica utc ultramicrotome, collected on nickel grids coated with 0.3% formvar (sigma) and stained with 1% phosphotungstic acid and 5% uranyl acetate solutions. sections were examined using a joel em 1010 transmission electron microscope. statistical analysis results of the pot experiment were statistically analyzed using the student’s t-test to evaluate the differences in biomass production (leaves, stems and roots) between control plants (non-inoculated) and plants inoculated with four different fungal strains. two-factor anova and newman–keuls multiple range test (p ≤ 0.05; for comparison of means) were used to compare the effects of inoculation on biomass production of pvy+ and pvy− plants. all statistical analyses were performed using statistica for windows, version 7.0. results impact of mycorrhizal, saprotrophic and pathogenic fungi on the growth of pvy-infected and non-pvy-infected potato cv. pirol plants fungal strains used for inoculation differed in their effects on pvy-infected and nonpvy-invected plants. only the mycorrhizal fungus g. intraradices increased the growth of pvy(+) plants compared to the uninoculated control plants. significant differences were recorded for the dry weight of leaves and stems (fig. 1a–c). the other fungal strains either inhibited (t. viride; leaves and roots) or did not affect (t. harzianum and c. coccodes) the growth parameters of pvy(+) potato plants. remarkably, no growth promotion of pvy(−) plants by the mycorrhizal treatment was observed. the non-mycorrhizal t. viride, t. harzianum and c. coccodes fungal strains decreased the growth of pvy(−) potato plants, especially the development of leaves and stems. however, the two-way anova statistical analysis (tab. 1) only showed a significant effect of pvy infection in respect to the dry weight of stems. dry weight of leaves and stems, but not of roots, was significantly affected in plants inoculated with all selected fungal strains. however, 295© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization statistically significant reduction of dry weight of leaves and stems was only observed for plants treated with saprophytic strains t. viride or t. harzianum. fig. 1 impact of glomus intraradices, trichoderma viride, t. harzianum and colletotrichum coccodes inoculation on the dry weight of leaves (a), stems (b) and roots (c) of pvy-infected (pvy+) or virus-free (pvy−) plants of potato cv. pirol (student test, n = 3, ±sd). s – stimulation; i – inhibition. leaves shoots roots ms effect f p ms effect f p ms effect f p 1. virus 0.0886 2.8084 0.1093 0.1658 4.9079 0.0385* 0.2253 0.3071 0.5856 2. inoculation 0.2698 8.5557 0.0003* 0.1556 4.6073 0.0084* 1.1529 1.5712 0.2206 3. interaction 1 × 2 0.0789 2.5029 0.0749 0.0633 1.8745 0.1544 0.6170 0.8409 0.5155 newman–keuls test 1. virus pvy(−) 0.4470 a 0.5057 a 1.2470 a pvy(+) 0.5557 a 0.6543 b 1.4203 a 2. inoculation control 0.6692 b 0.5458 ab 1.1025 a g. intraradices 0.6950 b 0.7633 b 1.8433 a t. viride 0.1917 a 0.4717 ab 0.7350 a t. harzianum 0.3833 ab 0.3917 a 1.6450 a c. coccodes 0.5675 b 0.7275 b 1.3425 a tab. 1 the effects of interaction between two factors: infection of plants with virus particles [(+pvy) and (−pvy)] and inoculation of plants with fungi (glomus intraradices, trichoderma viride, t. harzianum, colletotrichum coccodes) on the biomass production (leaves, stems and roots) in plants of potato cv. pirol (anova2; p ≤ 0.05). 296© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization analysis of the stress level after pvy infection and fungal association – determination of hydrogen peroxide (h 2 o 2 ) level after fungal inoculation, pvy(+) and pvy(−) plants differed in the h2o2 concentrations when compared with control plants. the highest level of hydrogen peroxide was recorded for leaves of plants inoculated with t. viride and c. coccodes (fig. 2a). for both above types of inoculation, leaves of pvy(+) plants revealed 1.4–2.4-fold higher amounts of h2o2 than leaves of pvy(−) plants. the elevated h2o2 level in leaves of t. viride-inoculated plants was correlated with a significant decrease in the leaf biomass (fig. 1a and tab. 1). such a relationship was not observed after inoculation of plants with c. coccodes. the plants inoculated with g. intraradices and t. harzianum only showed a slightly higher h2o2 level compared to leaves of non-inoculated control plants. the h2o2 profile in the roots differed from that observed in the leaves (fig. 2b). both pvy(+) and pvy(−) plant roots inoculated with g. intraradices exhibited similar h2o2 concentrations compared to the non-inoculated variant. inoculation with t. harzianum resulted in a decrease in h2o2 level in the roots of pvy(−) plants, whereas the pvy(+) plants showed the same level of this compound as did the control plants. inoculations with t. viride and c. coccodes resulted in a similar way, increasing the level of h2o2 in pvy(−) plant roots and decreasing the level of this compound in pvy(+) plant roots (fig. 2b). fig. 2 impact of glomus intraradices, trichoderma viride, t. harzianum and colletotrichum coccodes inoculation on the production of hydrogen peroxide in leaves (a) and roots (b) of pvy(+) and pvy(−) plants of potato cv. pirol. the level of hydrogen peroxide is presented as a percentage of stimulation or inhibition in relation to the control (100%; mean value ±sd). 297© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization correlation of plant growth parameters and h 2 o 2 level growth parameters (dry weight of leaves, stems and roots) of pvy(+) cv. pirol plants were analyzed in relation to the h2o2 levels in leaves and roots. a significant negative correlation was found between the dry weight of roots and h2o2 level in the leaves (r = −0.94, p = 0.02; fig. 3a). a similar relationship was observed for the dry weight and h2o2 level in leaves, but it was not statistically significant (data not shown). in contrast, a significant positive correlation between dry weight and h2o2 level in the roots of pvy(−) plants was found (fig. 3b). remarkably, the level of h2o2 in leaves or roots had no impact on the biomass of stems. this observation suggests that stems are not involved in the reaction of plants. perhaps, this organ plays a role as a transmitter of signals between the aboveand below-ground parts of the plants. observation of pvy particles in hyphae of g. intraradices the results of ultrastructural analysis confirmed the presence of hyphae of g. intraradices penetrating the intracellular spaces of potato roots (fig. 4a–c), thus confirming an effective colonization process. this symbiotic fungus formed unique structures, such as arbuscules and vesicles in the roots. in addition, numerous bacterial cells were observed inside the plant cells (fig. 4c,d). they were mostly concentrated around the fungal coils, or (rarely) distributed along the cell wall of the host plant. fig. 4a and fig. 4b depict structures similar to virus particles visible inside the hyphae forming arbuscules. fig. 4d shows the concentration of described above virus-like particles around bacterial cell clusters. discussion mutual relationships between two (or more) pathogens and/or symbionts in the same host plant are hardly to assess in their total diversity and significance. therefore the present study is a pilot model to indicate general processes in this interaction by a selected combination of one potato variety with one virus and four fungal strains only. we demonstrate significant stimulation of growth of pvy-infected potato plants inoculated with the mycorrhizal g. intraradices fungus. our results are opposite to the observations of sipahioglu et al. [14] who revealed significantly greater reduction in plant height and fig. 3 correlation of growth parameters of pvy(+) and pvy(−) plants of potato cv. pirol with the h2o2 level in leaves and roots. 298© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization root development in pvy-infected potato plants inoculated with g. intraradices. it is known that potato plants are colonized by arbuscular fungi can promote plant growth (e.g., bayrami et al. [15]). however, in the present study, a significant effect of mycorrhization on the growth of healthy potato plants was only observed in the roots. it is well known that mycorrhiza formation can lead to growth promotion by increased supply of water and nutrients [16]. furthermore, promotion of plant growth might favors virus multiplication. increased multiplication of plant viruses by mycorrhizal colonization of the host plants was revealed previously [17]. yet, only one report [18] showed the enhanced disease severity compared with non-mycorrhizal controls. based on investigations involving three viruses (tomato acuba mosaic virus, potato virus x, arabis mosaic virus) in three different hosts (tomato, petunia, strawberry), daft and okusanya [17] revealed that the amount of extractable virus particles is higher for mycorrhizal than non-mycorrhizal plants. they suggested that this is due to the high phosphate level in mycorrhizal plants. enhanced virus multiplication was also observed for non-mycorrhizal plants grown under increased supply of soluble phosphate [17]. besides, colonization of roots by mycorrhizal fungi was reported to increase the ability of plants to activate defense processes during pathogen infection [9]. it has been shown that physical contact of arbuscular fungi with fig. 4 electron microscope (tem) micrographs of the roots of potato cv. pirol infected with pvy particles and inoculated with glomus intraradices. am – mycelium of arbuscular fungus; cw – cell wal; v – vesicules; b – bacteria. viral-like particles are marked with arrows. bar: 500 nm. 299© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization plant roots can affect the expression of genes involved in the defense system [7,9]. during the infection process, cells of mycorrhized plants were determined to exhibit an increased level of antioxidant enzymes and pathogenesis-related (pr) proteins [9]. furthermore, mycorrhized plants were found to express increased resistance against pathogens, e.g., capsicum annuum against infection with phytophthora capsici [19], or medicago truncatula against the bacterial pathogen xanthomonas campestris [7]. meanwhile, the number of reports supporting the existence of a protective effect of mycorrhizal fungi on plants infected with viruses is still scarce. beside mycorrhizal fungi, several saprotrophic fungal species like trichoderma spp. show plant growth promoting capacities, such as synthesis of plant hormones and vitamins as well as increased intensity of nutrient uptake from soil (e.g., n and p) [6]. however, in our experiments both t. harzianum and t. viride strains significantly inhibited the growth of pvy(+) and pvy(−) control plants. the observed growth inhibition of potato plants by trichoderma strains might have been caused by temporal nutrient competition or plant species-specific effects. such differences, due to colonization with trichoderma strains, have been reported earlier for tomato and cucumber [10]. so far there is no knowledge on the resistance of potato to black dot caused by the c. coccodes fungus. here, the influence of this pathogen on virus-infected potato plants was investigated for the first time. in our study, c. coccodes significantly decreased the dry weight of leaves of pvy(−) plants, but had no significant effect on pvy(+) plants. as c. coccodes infection starts in below-ground plant organs and spreads into the above-ground stems [20], we suggest that pvy infection might delay the dissemination of the fungus into the upper plant parts, e.g. by stronger or earlier activation of defense mechanisms in the plant. in the present study, slightly increased concentrations of foliar hydrogen peroxide (h2o2) were noted in mycorrhized plants compared to the non-mycorrhizal control. the mechanism by which the mycorrhizal fungi induce resistance in plants is still unclear [19]. it is known that some of the genes responsible for the formation of mycorrhizal symbiosis are activated during pathogen infection [7,9]. in both cases, changes in the synthesis of ros were observed [17]. vasse et al. [21] revealed that ros are generated in response to all plant-invading microorganisms (also symbiotic), and react with the defensive response as long as the symbiont is not recognized as such [21]. moreover, it was observed that development of the mutualistic association between the fungal endophyte – epichloë festucae, and its host plant (lolium perenne) requires production of superoxide or hydrogen peroxide by a fungal nadph oxidase, while inactivation of this gene changes the fungi-host interaction from mutualistic to antagonistic [22]. the level of h2o2 in mycorrhized plants was not affected by pvy infection, although various types of abiotic and biotic stress can lead to an uncontrolled increase of ros formation, leading both to damage of biomolecules and activation of defense mechanisms [13]. in the cells of mycorrhized plants, the h2o2 level after inoculation with a pathogen may decrease compared to non-mycorrhized plants [19]. literature data shows that the highest h2o2 level in mycorrhized plants infected with pathogens can be observed already after 6 h, while in non-mycorrhized plants after 12 h [19]. this indicates a faster induction of defense mechanisms in mycorrhized plants. since in our study the level of h2o2 was tested 10 weeks after plant inoculation, any potential factors contributing to induction of resistance would have already acted. the effect of mycorrhization of solanum lycopersicum 300© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization plants on the infection process with tomato spotted wilt virus (tswv) suggests that mycorrhizae may cause suppression of plant response to viral infection [23]. mycorrhizal symbiosis was shown to decrease expression of genes activated during viral infection and reduce visual symptoms in long-term [23]. similarly, in our study growth promotion of virus-infected plants in mycorrhizal symbiosis was observed. beside the mycorrhizal effects, it was determined that inoculation of potato plants with t. viride significantly stimulated the formation of reactive oxygen species; yet this was not correlated with plant growth promotion. participation of trichoderma sp. in enhancement of plant resistance against pathogens by activating the defence response was earlier shown by nawrocka et al. [10]. trichoderma spp. can affect other soil microorganisms, e.g. by antibiosis [4]. some antibiotics synthesized by microorganisms may be active against viruses [6,24]. as was shown previously, some peptide antibiotics synthesized by trichoderma sp. can significantly reduce the development of tmv in tobacco plants (nicotiana tabacum) [24]. it was suggested that an increased level of reactive oxygen species, like we observed in our study, might indicate that substances synthesized by fungi induce immunity in the infected plants. infection of s. tuberosum l. with the pathogenic c. coccodes significantly increased the h2o2 level in the leaves of both pvy(+) and pvy(−) plants compared to the non-inoculated control. this effect is suggested to be due to the activation of the ros synthesis in response to pathogen infection [25]. specifically, this could be caused by secretion of ammonium (as the source of deamination of amino acids), which in turn can lead to alkalization of the environment from ph 4.2 to 7.9, during all growth stages of colletotrichum spp. [26]. ammonium secreted by c. coccodes can enhance ros accumulation, promoting cell death and fungal virulence [26]. moreover, it was shown that ammonium indirectly activates the transport of various host solutes, modulates concentrations of host cytosolic protons [27], and alters the membrane flux processes [28]. as the level of h2o2 in our experiment was tested 10 weeks after plant inoculation, it could be that the highest accumulation of h2o2 in plants appeared much earlier. literature data reports that increased levels of h2o2 in plants after contact with fungi, can initiate adequate defense mechanisms, called “fast reaction” [29]. in the absence of an appropriate fungal signal, a prolonged infection process and consistently high h2o2 concentrations induce a permanent stress reaction. in such a situation, a microorganism is recognized as incompatible [30]. based on the gathered data, we speculate that the specific features of each fungal strain rather than colonization per se play a key role in the plant response related to h2o2. moreover, it seems that the antioxidant systems lead to different colonization strategies depending on the fungal strain involved. a constantly high h2o2 concentration is probably the main toxic factor during stress conditions that contributes to the decreased biomass production [31]. results of our study confirm this supposition. high h2o2 levels in pvy(+) compared to virus-free plants suggest a permanent stress reaction induced by the viral particles in plant tissues. in such plants, the virus can move to its upper parts, e.g., leaves. in effect, high h2o2 levels that were noted may have been responsible for the reduction of root biomass. in the absence of pvy particles in plant tissues, fungi remained the only infectious agent, which had no detectable influence on plant growth and biomass production. the observed significant correlation of increased biomass with increased h2o2 levels in the roots might be a consequence of intensive cell divisions in this plant organ [29]. 301© the author(s) 2014 published by polish botanical society acta mycol 49(2):291–303 thiem et al. / response of pvy infected potato to fungal colonization beside the observed physiological effects in plants, ultrastructural analysis suggests that mycorrhizal fungi may be involved in the transfer of virus particles within the plant. xavier and boyetchko [32] imply that mycorrhizal fungi do not play a role as viral vectors and do not interact with viruses. however, arbuscular mycorrhizal (am) symbioses were associated with enhanced susceptibility to viruses in earlier reports (reviewed by whipps [33]). sipahioglu et al. [14] revealed that the inoculation of pvy-infected potato plants with g. intraradices reduced vegetative development but increased viral activity considerably. transmission of certain viruses by fungal vectors has been previously suggested by hollings and stone [34]. however, no sufficient data supporting this hypothesis has been provided so far. the current insufficient knowledge about the epidemiology of diseases caused by plant viruses are in part a consequence of research activities directed solely on the plant-virus interactions, omitting the presence of other plant-associated microorganisms. our study suggests the presence of pvy particles in arbuscules formed in the roots of s. tuberosum inoculated with g. intraradices. however, future immunodetection analysis has to confirm this observation, focusing on this specific aspect of the model system. we suppose that beside the growth promotion of virus-infected host plants, the mycorrhizal g. intraradices might be a potential vector for plant viruses, which would be highly significant for potato breeding systems and seed potato production, but has to be tested in a subsequent investigation focusing this aspect. glomus intraradices might protect s. tuberosum l. also indirectly by accompanying beneficial bacteria (fig. 4c). fig. 4d presents the concentration of bacteria in the vicinity of virus-like particles. it is known that endophytic bacteria may induce immunity against some pathogens [8]. their specific significance in pvy(+) potato plants also still needs further investigation. it is widely known that studies on multi-component interactions are difficult and laborious, and do not always yield unequivocal results. in our experiment mycorrhizal fungus g. intraradices significantly stimulated the growth of pvy-infected potato plants, whereas two saprophytic trichoderma fungi and a pathogenic strain of c. coccodes had either no effect or inhibitory effect on virus-infected plants. the highest level of h2o2, an indicator of biotic stress, was recorded in plants inoculated with t. harzianum and c. coccodes, which caused the highest inhibition of plant growth. we are aware that the plant material (one cultivar of potato only) is not sufficient to draw generalized conclusions. however, the present model experiment revealed the potential transfer function of fungal hyphae for both pathogens (like viruses) and plant growth promoting endophytes in potato plants. the verification of these projections is the future challenge of the ongoing research. acknowledgments this investigation was done in frame of cost action fa1103. authors’ contributions the following declarations about authors’ contributions to the research have been made: performed pot experiment and biomass analysis: dt; performed biochemical experiments and participated in the manuscript preparation: asj, cb, km; prepared tem analysis: kn; supervised the research design and wrote the paper: kh. references 1. syller j, kaliciak a. rośliny 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(vol 2). 33. whipps jm. prospects and limitations for mycorrhizas in biocontrol of root pathogens. can j bot. 2004;82(8):1198–1227. http://dx.doi.org/10.1139/b04-082 34. hecht ei, bateman df. non-specific acquired resistance to pathogens resulting from localised infections by thielaviopsis basicola on viruses in tobacco leaves. phytopathology. 1964;54:523–530. http://dx.doi.org/10.1046/j.1365-313x.1993.04030555.x http://dx.doi.org/10.1046/j.1365-313x.1993.04030555.x http://dx.doi.org/10.1016/j.plantsci.2010.10.002 http://dx.doi.org/10.1094/mpmi-05-11-0116 http://dx.doi.org/10.1111/j.1574-6968.2010.02135.x http://dx.doi.org/10.1111/j.1574-6968.2010.02135.x http://dx.doi.org/10.1016/j.micron.2009.12.004 http://dx.doi.org/10.1016/j.micron.2009.12.004 http://dx.doi.org/10.1094/mpmi-22-12-1484 http://dx.doi.org/10.1078/0176-1617-0774 http://dx.doi.org/10.1104/pp.106.079467 http://dx.doi.org/10.1094/mpmi.2001.14.1.86 http://dx.doi.org/10.1094/mpmi.2001.14.1.86 http://dx.doi.org/10.1104/pp.106.079079 http://dx.doi.org/10.1139/b04-082 abstract introduction material and methods plant and microbial material pot experiment determination of hydrogen peroxide (h2o2) level electron microscopy analysis statistical analysis results impact of mycorrhizal, saprotrophic and pathogenic fungi on the growth of pvy-infected and non-pvy-i analysis of the stress level after pvy infection and fungal association determination of hydrogen correlation of plant growth parameters and h2o2 level observation of pvy particles in hyphae of g. intraradices discussion acknowledgments authors’ contributions references 2014-12-31t17:46:25+0000 polish botanical society 2014-09-06t20:22:46+0200 polish botanical society 2014-11-20t23:16:29+0000 polish botanical society 2014-11-20t23:20:01+0000 polish botanical society 2014-11-20t23:13:10+0000 polish botanical society 2014-11-20t23:17:58+0000 polish botanical society 2014-11-20t23:18:23+0000 polish botanical society 2014-11-20t23:17:20+0000 polish botanical society threatened and protected macromycetes in the wkrzańska forest stefan friedrich department of botany and nature protection, university of agriculture słowackiego 17, pl 71 434 szczecin; sfriedrich@agro.ar.szczecin.pl f r i e d r i c h s.: threatened and protected macromycetes in the wkrzańska forest. acta mycol. 41(2): 229 240, 2006. eightysix interesting species of macromycetes, including fungi red listed in poland (84 species) and fungi protected in poland (11 species), observed in the wkrzańska forest are described. a total of 460 species of macromycetes were recorded during mycosociological studies conducted in the period between 2003 and 2005. key words: macromycetes, wkrzańska forest, threatened species, protected species, red list of fungi introduction the wkrzańska forest is a vast complex of ca. 1 550 km2 divided between poland and germany (fig. 1). the polish part, covering 340 km2 (z a r ę b a 1986), comprises the wkrzańska plain and the northern part of the szczecińskie hills (k o n d r a c k i 1998). leucobryo-pinetum and querco roboris-pinetum phytocoenoses of various age dominate in the central and northern plain parts of the wkrzańska forest. broadleaved and mixed tree-stands of the luzulo pilosae-fagetum and fago-quercetum petraeae associations occur in its western and southern parts on moraine hills. small sections, scattered over the study area, are overgrown with the following associations: ribeso nigri-alnetum, betulo pendulae-quercetum roboris, fraxino-alnetum, vaccinio uliginosibetuletum pubescentis. the forests are administered by the trzebież forest district. macromycetes in the wkrzańska forest had not been examined before. only 5 species of macromycetes common in poland collected during studies on poplar mycotrophy have been reported from the area (d o m i n i k , i h n a t o w i c z 1979): laccaria laccata, laetiporus sulphureus, polyporus squamosus, trametes versicolor and vascellum pratense. on the other hand, the mycobiota of the adjacent regions has been investigated in relative depth: the goleniowska forest to the east of the study area (f r i e d r i c h 1984, 1985a, 1985b, 1997) and the city of szczecin to the south (f r i e d r i c h 1987; f r i e d r i c h , o r z e c h o w s k a 2002). systematic mycological studies in the wkrzańska forest commenced in 2002. the occurrence and acta mycologica vol. 41 (2): 229-240 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 230 s. friedrich fig. 1. localization of macromycetes observation plots in wkrzańska primeval forest: 1 observation plots (sites), 2 numbers of the grid atpol squares, 3 forests boundary, 4 state boundary. threatened and protected macromycetes 231 the distribution of threatened and protected macromycetes are presented below. the participation and the role of macromycetes in the forest communities of the wkrzańska forest will be discussed separately. analysis results will be used to compile a regional list of threatened fungi in pomerania. methods systematic mycosociological research was conducted at 21 permanent observations plots, 400 m2 each, set up in forest communities. community sociology was determined by the present author. supplementary studies, that is individual mycological and phytosociological observations at additional 64 plots, also 400 m2 each, were carried out throughout the periods of increased fungal fruiting. the sites were situated in the atpol grid squares (tab. 1). fruiting abundance was evaluated in the m o s e r ’ s scale (1949). the growth manner of the fruit-bodies on the substrate was also identified. mycological observations were additionally performed outside the forest areas (roadsides, forest tracts, glades, etc.) using the itinerary method, fungal species were recorded, and occurrence conditions were noted. studies were continued over 3-4 years in permanent plots which were examined 5 times per year on average. fungal taxa of red-listed threatened species (wo j e w o d a , ł a w r y n o w i c z 1992) and protected species (dz. u. 2004.168.1765) were included in the study. results and discussion the occurrence of 460 fungal species belonging to all ecological groups were recorded during studies conducted at 85 sites between 2002 and 2005. a high percentage of mycorrhizal fungi which constitute 38% of the recorded macromycetes should be stressed. the participation of other groups is as follows: lignicolous fungi – over 30%, litter-inhabiting and bryophilous fungi – over 23%, humicolous saprobes – over 6%, plant and fungal parasites – less than 3%. many of these taxa are threatened or rare not only in poland. the occurrence of 86 interesting species collected between 2002 and 2005 is described below. a total of 84 red-listed species threatened in poland and 11 species protected in poland were recorded in the mycobiota of the wkrzańska forest. one species is classified as extinct (ex), 4 species are classified as endangered (e), 16 species are classified as vulnerable (v), 31 species are classified as rare (r), and 32 species are classified as indeterminate (i). some threatened species are protected, and only 2 protected species (langermannia gigantea, meripilus giganteus) are not red-listed. the nomenclature and systematics of the orders follow k i r k et al. (2001), and the nomenclature of the species follows many sources cited by wo j e w o d a (2003), mostly: a n t o n í n & n o o r d e l o o s (1997), h a n s e n & k n u d s e n (1992, 1997, 2000), k r e i s e l (1987), k r i e g l s t e i n e r g . j . (1991), k r i e g l s t e i n e r l . (1999). species within orders are reported alphabetically. threat category and protection category as well as the substrate type are given for each species, and the plant community in which the species occurs is specified. the following data are also provided: quantity range of the occurrence of fruit-bod232 s. friedrich ta b l e 1 sites of macromycetes in the wkrzańska forest no atpol forest district forest division plant association no atpol forest district forest division plant association 1 ba73 tanowo 813c lp 39 ba53 pienice 110i qrp 2* ba73 tanowo 771g lp 40 ba53 pienice 148b pine greenwood 3 ba72 tanowo 772 fqp 41 ba52 pienice 115a lp 4* ba63 tanowo 718g lp 42 ba52 pienice 83g lpf 5 ba63 tanowo 718f qrp 43 ba52 pienice 82l qrp 6 ba63 tanowo 718b pine greenwood 44 ba62 tatynia 666d lpf 7 ba62 tanowo 669d lp 45* ba62 tatynia 667b lpf 8 ba62 tanowo 670j pine greenwood 50 ba62 tatynia 577g lp 9 ba63 tusznica 589a lp 51 ba62 tatynia 531c qrp 10 ba63 tusznica 564b lp 52 ba62 zalesie 532b qrp 11 ba63 tusznica 516l lpf 53 ba62 podymin 440h lp 12 ba63 tusznica 542f lpf 55* ba62 podymin 444f lpf 14 ba62 tusznica 518g lp 58 ba62 zalesie 490a lpf 16 ba52 myślibórz 260d lpf 59 ba52 podymin 384f lp 17 ba52 myślibórz 174f qrp 63 ba52 myślibórz 299i lpf 18* ba52 nowe warpno 66j lp 64 ba52 myślibórz 257b lp 19 ba52 nowe warpno 66h lp 65 ba52 myślibórz 293b lp 20 ba52 nowe warpno 39g pine greenwood 66 ba52 mazańczyce 197c lp 21 ba52 nowe warpno 23d lp 67* ba73 siedlice 827c lpf 22 ba52 nowe warpno 23f qrp 68* ba73 siedlice 826d fqp 23 ba52 nowe warpno 61ab bpqr 69 ba73 siedlice 807j lpf 24* ba52 pienice 89d lpf 71 ba73 siedlice 801g qrp 25 ba52 pienice 89c fqp 72* ba63 siedlice 801c fqp 26* ba52 pienice 86h fqp 77 ba72 tanowo 751m lp 27 ba52 pienice 119d lpf 78 ba51 podymin 342d lpf 28 ba73 tanowo 792n qrp 79 ba62 dobra 693d lpf 29 ba63 tusznica 565a qrp 81 ba72 tanowo 797a lp 30* ba63 tusznica 541c rna 82 ba52 drogoradz 360c raised bog 35 ba53 mazańczyce 270g lpf 83 ba42 nowe warpno 4d bpqr 36* ba53 mazańczyce 271d bpqr 84 ba62 zalesie 491d pine greenwood 37* ba53 mazańczyce 226d qrp 85 ba63 tanowo 720b pine greenwood 38* ba53 mazańczyce 226b rna threatened and protected macromycetes 233 ies in the m o s e r ’ s scale (1949), record date and a list of localities recorded between 2002 and 2005 preceded by their number in parentheses. the quantity is not given for species producing permanent fruit-bodies. the latin plant nomenclature follows m i r e k et al. (2002) and that of communities follows m a t u s z k i e w i c z (2001). abbreviations: threat categories in poland (w o j e w o d a , ł a w r y n o w i c z 1992): pl ex extinct, pl e endangered, pl v vulnerable, pl r rare, pl i indeterminate; §§ fully protected species, § partially protected species (regulation of the minister of the environment of 9th july 2004); rna ribeso nigri alnetum, lp leucobryo pinetum, qrp querco roboris pinetum, bpqr betulo pendulae quercetum roboris, fqp fago quercetum petraeae, lpf luzulo pilosae fagetum; + 5 quantity range of fruit bodies according to m o s e r (1949); jan. january, aug. august, sept. september, oct. october, nov. november, dec. december; * permanent observation plot. ascomycetes helotiales ascotremella faginea (peck) seaver pl-v; on a fallen branch of fagus sylvatica; lpf; 1; july 2004; site (1): 69, (fig. 2). bulgaria inquinans (pers.: fr.) fr. pl-r; on logs and fallen branches of quercus petraea and q. robur; bpqr, qrp; 3; oct. 2004, nov. 2005; sites (2): 36*, 51. chlorencoelia versiformis (pers.: fr.) dixon pl-i; on a log of quercus robur; bpqr; 1; oct. 2004; site (1): 83. basidiomycetes dacrymycetales calocera furcata (batsch.: fr.) fr. pl-i; on logs and fallen branches of pinus sylvestris; lp, qrp; 2-4; july-nov. 2002-2005; sites (12): 7, 8, 9, 12, 19, 39, 40, 43, 53, 69, 84, 85. tremellales tremella foliacea pers. pl-i; on trunks and logs of betula pendula; lp; +-1; july 2004, oct. 2005; sites (3): 4*, 14, 64. tremella globospora d. a. reid pl-i; on old stromata of pyrenomycetous fungi growing on woods; pine greenwood; 2; oct. 2002; site (1): 8, (fig. 3). agaricales amanita citrina var. alba (gillet) gilbert pl-i; on the ground under pinus sylvestris; lp; +-1; oct. 2005; sites (2): 2*, 65. amanita virosa (fr.) bertillon pl-v; on the ground under pinus sylvestris; lp; 1; sept. 2003; site (1): 2*. 234 s. friedrich arrhenia retiruga (bull.: fr.) redhead pl-i; on mnium hornum moss; rna: 2.2; nov. 2004; site (1): 38*, (fig. 4). arrhenia spathulata (fr.: fr.) redhead pl-i; on tortula ruralis moss; roadside; aug. 2004; site (1): near karszno. clitocybe candicans (pers.: fr.) p. kumm. pl-i; on the litter; rna, qrp, fqp, lpf; 2.2; sept.-nov. 2003-2005; sites (4): 3, 24*, 29, 38*. clitocybe hydrogramma (bull.: fr.) p. kumm. pl-r; on the broadleaved litter; lpf; nov. 2004; site (1): 16. coprinus picaceus (bull.: fr.) gray pl-v; on the ground; lpf; +-1; sept.-oct. 2004, 2005; sites (2): 16, 67*. cortinarius armeniacus (schaeff.: fr.) fr. pl-i; on the ground; lp; 1; sept. 2004; sites (2): 41, 81. cortinarius orellanus fr. pl-i; on the ground; qrp; 1-2; july 2004; site (1): 17. cystodermella granulosum (batsch: fr.) harmaja pl-r; on the ground; qrp, +-1; sept.-oct. 2004; sites (2): 17, 28. entoloma juncinum (kühner. & romagn.) noordel. pl-i; on the litter; fqp; 1; july 2004; site (1): 25. entoloma papillatum (bres.) dennis pl-i; on the ground; qrp; 1; oct. 2005; site (1): 51. fayodia maura (fr.) singer pl-i; on a charred branch of pinus sylvestris; lp; 1; oct. 2002; site (1): near 85. fistulina hepatica (schaeff.): fr. pl-v, §§; on a trunk of quercus robur; 1; sept. 2005; site (1): near 45*. galerina paludosa (fr.) kühn. pl-i; among sphagnum spp.; raised bog on lake piaszynko; 2; july 2004; site (1): 82. galerina sphagnorum (pers.: fr) kühner pl-i; among sphagnum spp.; raised bog on lake piaszynko; 2; july 2004; site (1): 82. galerina triscopa (fr.) kühner pl-i; on a log of fagus sylvatica; lpf; 1; oct. 2004; site (1): 27. gymnopus putillus (fr.: fr.) antonín, halling & noordel. pl-i; on the litter; qrp; 1; aug. 2004, sept. 2005; site (1): 22. inocybe acuta boud. pl-r; on the ground; qrp; 1; july 2004; site (1): 43. inocybe calospora quél. pl-r; on the ground; rna; july 2004; site (1): 30*. langermannia gigantea (batsch: pers.) rostk. – §§; on the ground; july 2004, sept. 2005; site (1): the świdwie ornithological station, (fig. 5). lepiota tomentella j. e. lange pl-i; on the ground; lp; 1; oct. 2003; site (1): 66. lyophyllum palustris (peck) singer pl-i; among sphagnum spp.; raised bog on lake piaszynko; 2; sept. 2004; site (1): 82. macrolepiota procera (scop.: fr.) singer pl-i; on the ground; qrp, lpf, pine greenwood, roadside; +-1; sept.-nov. 2002-2005; sites (5): 5, near 6, 20, 51, 64. macrolepiota rhacodes (vittad.) singer pl-i; on the ground; qrp, forest roadside; 1; july 2004; sites (2): near 28, 71. marasmiellus foetidus (sowerby: fr.) antonín, halling & noordel. pl-r; on fallen twigs of fagus sylvatica; lpf; 2; nov. 2005; site (1): 42. mycena adonis (bull.: fr.) gray pl-r; on the litter and on fallen twigs of pinus sylvestris; lp; 2; oct. 2004; site (1): 81. threatened and protected macromycetes 235 mycena atroalba (bolt.: fr.) gillet pl-r; on a log of pinus sylvestris, 2; sept. 2003; site (1): 28. mycena belliae (johnst.) p. d. orton pl-r; on culms of phragmites australis; phragmitetum australis; 2-3; sept.-nov. 1999-2004 (leg. k. olszanowski); site (1): lake świdwie. mycena crocata (schrad.: fr.) p. kumm. pl-r; on fallen twigs of fagus sylvatica; lpf; 2; nov. 2005; sites (2): 58, 67*. mycena olivaceomarginata (massee) massee pl-r; on the coniferous litter; qrp; 2; sept. 2004; site (1): 71. mycena pelianthina (fr.) quél. pl-i; on the broadleaved litter; lpf; 1-2; july 2004, nov. 2005; sites (3): 24*, 45*, 63. mycena purpureofusca (peck) sacc. pl-v; on a stump of pinus sylvestris; lpf; 2; oct. 2004; site (1): 45*. mycena septentrionalis maas geest. pl-r; on the litter; lp; 1; sept. 2004; site (1): 59. mycena viridimarginata p. karst. pl-v; on a stump of fagus sylvatica; lpf; july 2004; site (1): 44. omphaliaster asterosporus (j. e. lange) lamoure pl-e; on the litter among mosses; lp; 1-2; nov. 2005; sites (3): 7, 9, 64. oudemansiella mucida (schrad.: fr.) höhn. pl-v; on trunks, logs and fallen branches of fagus sylvatica; lpf; 2-3; july-nov. 2003-2005; sites (6): 24*, 35, 45*, 55*, 67*, 79, (fig. 6). pleurotus dryinus (pers.: fr.) p. kumm. pl-r; on stumps of alnus glutinosa; rna; 1; nov. 2003; site (1): 30*. pluteus atromarginatus (singer) kühner pl-v; on stumps of pinus sylvestris; lp; +1; july 2004, sept.-oct. 2005; sites (3): 37*, 50, 64. psathyrella gracilis (fr.: fr.) quél. pl-i; on the ground; lpf; 1; oct. 2003; site (1): 35. psilocybe elongata (pers.: fr.) j. e. lange pl-v; among sphagnum spp.; raised bog on lake piaszynko; 1; oct. 2004; site (1): 82. psilocybe polytrichi (fr.: fr.) pears. & dennis pl-r; on the litter among mosses; lp; oct. 2002, nov. 2004, sept. 2005; sites (4): 14, 19, 64, 77. psilocybe uda (pers.: fr.) gillet pl-r; among sphagnum spp.; raised bog on lake piaszynko; 2; oct. 2005; site (1): 82. rhodocollybia prolixa (hornem.: fr.) antonín & noordel. var. distorta (fr.) antonín, halling et noordel. pl-i; on the litter; lp, qrp; 1-2; sept. 2004, oct. 2005; sites (2): 21, 29. tricholoma equestre (l.: fr.) p. kumm. ss. lato pl-i; on the ground under pinus sylvestris; lp, pine greenwood; 1-2; oct. 2003, nov. 2005; sites (2): 2*, 66. tricholoma pessundatum (fr.: fr.) quél. pl-i; on the ground under pinus sylvestris; lp; 1; sept. 2004; site (1): 77. tricholoma sejunctum (sowerby: fr.) quél. pl-i; on the ground under fagus sylvatica and quercus petraea; fqp; 1; oct. 2005; site (1): 54. 236 s. friedrich boletales boletus edulis bull.: fr. pl-v; on the ground under: fagus sylvatica, pinus sylvestris, quercus robur; qrp, lpf, july, oct. 2004, 2005; +-1; sites (4): 5, 51, 55*, 58. boletus pinophilus pilát & dermek – pl-i; on the ground under pinus sylvestris; lp; +-1; july 2004, sept. 2005; site (1): 18*. boletus suspectus krombh. pl-e; on the ground under fagus sylvatica; lpf; 1; nov. 2005; site (1): 24*. gomphidius roseus (fr.) fr. pl-r; on the ground; lp; +: oct. 2004, 2005; sites (2): 7, 64, (fig. 7). gyrodon lividus (bull.: fr.) sacc. pl-r; on the ground; rna; 1; july 2004; site (1): 38*, (fig. 8). cantharellales cantharellus cibarius fr. pl-i; on the ground under pinus sylvestris; lp; 1-2; julyoct. 2003-2005; sites (5): 2*, 19, 41, 66, 77. clavulina rugosa (bull.: fr.) j. schröt. pl-r; on the ground; lp, pine greenwood; 1-2; sept. 2004; sites (2): 8, 19. hymenochaetales inonotus obliquus (pers.: fr.) pilát pl-r, §; on a trunk of betula pendula; lp; jan.dec. 2004-2005; sites (2): 10, 18*. phallales clavariadelphus fistulosus (holmsk.: fr.) corner pl-r; on fallen twigs of betula pendula; lp; 1-2; oct. 2002; sites (2): 2*, 4*, (fig. 9). geastrum fimbriatum fr. pl-r, §§; on the ground; fqp; 2; july 2004; site (1): 68*. geastrum rufescens pers.: pers. pl-e, §§; on the ground; fqp; 1; oct. 2003; site (1): 26*. phallus duplicatus bosc pl-e; on the ground; lpf; +; july 2004; site (1): 55*. polyporales datronia mollis (sommerf.: fr.) donk pl-i; on stumps and logs of fagus sylvatica; lpf; jan.-dec. 2004-2005; sites (3): 11, 16, 45*. ganoderma lucidum (m. a. curtis: fr.) p. karst. pl-r, §§; on a stump of fagus sylvatica; lpf; sept. 2004; site (1): 24*. ischnoderma benzoinum (wahlenb.: fr.) p. karst. pl-r; on a log of pinus sylvestris; lp; 1; sept. 2003, oct. 2005; site (1): 2*. ischnoderma resinosum (fr.) p. karst. pl-v; on a log of fagus sylvatica; lpf; 2; oct. 2005; site (1): 45*, (fig. 10). threatened and protected macromycetes 237 lentinus tigrinus (bull.: fr.) fr. pl-i; on a log of alnus glutinosa and on a fallen branch of betula pendula; rna, bpqr; 1-2; sept. 2003, oct. 2005; sites (2): 23, 38*. meripilus giganteus (pers.: fr.) p. karst. – §§; at the bases of stumps and living trunks of fagus sylvatica; lpf, fqp; july-aug. 2004, 2005; sites (7): 35, 45*, 58, 63, 67*, 72*, 78, (fig. 11). oligoporus ptychogaster (ludwig) r. & o. falck – pl-r; on a log and on the coniferous litter of pinus sylvestris; lp; 2; sept. 2004; sites (2): 2*, 64, (fig. 12). phaeolus schweinitzii (fr.: fr.) pat. pl-r; on trunks of pinus sylvestris; lp, qrp; nov. 2004; sites (2): 21, 37*. pycnoporus cinnabarinus (jacq.: fr.) p. karst. pl-r; on logs and fallen branches of betula pendula and fagus sylvatica; lp, fqp; jan.-dec. 2002-2004; sites (4): 25, 41, 50, 66, (fig. 13). sparassis brevipes krombh. pl-v, §§; on a fallen branch of fagus sylvatica; lpf; +; nov. 2005; site (1): 24*, (fig. 14). sparassis crispa (wulf.): fr. pl-r, §§; on roots of pinus sylvestris; lp, qrp; +-1; sept.-oct. 2003-2005; sites (6): 1, 9, 22, 37*, 51, 77, 83. russulales hericium coralloides (scop.: fr.) pers. pl-v, §§; on logs of fagus sylvatica; lpf; +1; sept.oct. 2005; sites (3): 16, 67*, 79 (fig. 15). lactarius chrysorrheus fr. pl-r; on the ground under quercus petraea and q. robur; fqp, qrp; 1-2; sept.-oct. 2004, oct. 2005; sites (3): 25, 52, 68*. lactarius deliciosus (l.: fr.) gray pl-v; on the ground under pinus sylvestris; lp; 1-2; oct. 2004, sept. 2005; site (1): 21. russula alutacea (pers.: fr.) fr. – pl-i; on the ground under fagus sylvatica; lpf, qrp; +-1; oct. 2003, sept. 2004; sites (3): 11, 52, 69. russula coerulea (pers.) fr. pl-ex; on the ground under pinus sylvestris; lp; 1; oct. 2004; site (1): 65. russula olivacea (schaeff.) fr. pl-r; on the ground under fagus sylvatica; lpf, fqp; 1-2; oct. 2003, sept. 2004, oct. 2005; sites (3): 27, 72*, 79. stereum subtomenosum pouzar pl-r; on logs of quercus petraea and fagus sylvatica; lpf, fqp; jan.-dec. 2004-2005; sites (3): 35, 44, 68*. thelephorales phellodon tomentosus (l.: fr.) banker pl-v; on the ground in the litter; lp; 1; oct. 2003; site (1): 19. sarcodon imbricatus (l.: fr.) p. karst. pl-v, §§; on the ground; lp; 1; oct. 2004; site (1): 2*. thelephora caryophyllea (schaeff.): fr. pl-r; on the ground; lp; 1; sept. 2004; site (1): 19. 238 s. friedrich conclusions as many as 50 species of the threatened and protected taxa are very rare and occur only at 1 locality in the wkrzańska forest. twenty-six species were recorded at 2-3 localities, 6 species – at 4-5 localities, and 3 species – at 6-7 localities (sparassis crispa, oudemansiella mucida, meripilus giganteus). among the taxa studied, calocera furcata, recorded at 12 localities, is the most common one in the wkrzańska forest. species previously not reported from pomerania occur in the wkrzańska forest. these are for instance: arrhenia retiruga, a. spathulata, ascotremella faginea, ischnoderma benzoinum, i. resinosum, omphaliaster asterosporus, phallus duplicatus, sparassis brevipes, tremella globospora. species very rare in poland, including taxa previously known from 1-3 localities in poland, occur in the studied mycobiota. these are: phallus duplicatus, known only from 1 site, arrhenia retiruga, known from 2 localities, and ascotremella faginea, russula coerulea and tremella globospora, known from 3 localities. it should be stressed that the two former species were previously recorded 70-90 years ago. it should be noticed that many species of macromycetes rare in poland, occurring at a few localities, are frequent or even common in germany, especially in its southern part (wo j e w o d a 2003; k r e i s e l 1987; k r i e g l s t e i n e r g . j . 1991). phallus duplicatus, whose locality in the wkrzańska forest is the second site in poland, is the best example. the taxon occurs at ca. 50 localities in germany, including the lands adjacent to the study area (meklenburg – western pomerania, brandenburg). other species in this group include: boletus suspectus, entoloma papillatum, inocybe acuta, ischnoderma resinosum, marasmiellus foetidus, russula coerulea and sparassis brevipes. as russula coerulea is recorded at the fourth locality in poland, the species should be reclassified from extinct (ex) to endangered (e), as suggested by wojewoda (2003). over 52% threatened and protected species were recorded only in sporadic plots and during itinerary observations while only nearly 20% were recorded exclusively in permanent plots. the results confirm the significance of observations conducted outside permanent plots for the investigation of the biodiversity of macromycetes in the study area. references a n t o n í n v. , n o o r d e l o o s e . 1997. a monograph of marasmius, collybia and related genera in europe. part 2: collybia, gymnopus, rhodocollybia, crinipellis, chaetocalathus, and additions to marasmiellus. libri bot. 17. ihw verlag, münchen. d o m i n i k t. , i h n a t o w i c z a . 1979. badania mikotrofizmu topól w uprawach doświadczalnych insty tutu badawczego leśnictwa na terenie woj. szczecińskiego. prace ibl. 544: 67 112. dz.u.2004.168.1765. rozporządzenie ministra środowiska z dnia 9 lipca 2004 r. w sprawie gatunków dziko występujących grzybów objętych ochroną. f r i e d r i c h s . 1984. mikoflora puszczy goleniowskiej. acta mycol. 20 (2): 173 208. f r i e d r i c h s . 1985 a. macromycetes na tle zespołów leśnych puszczy goleniowskiej. acta mycol. 21 (1): 43 76. f r i e d r i c h s . 1985 b (1987). charakterystyka ekologiczno fenologiczna macromycetes puszczy gole niowskiej. acta mycol. 21 (2): 143 164. f r i e d r i c h s . 1987. macromycetes szczecina. bad. fizjogr. nad pol. zach. 38, ser. b. 5 26. threatened and protected macromycetes 239 f r i e d r i c h s . 1997. macromycetes of the proposed nature reserve wilcze uroczysko olszanka in the odra estuary. acta mycol. 32: 239 255. f r i e d r i c h s ., 2004. wstępne wyniki badań macromycetes puszczy wkrzańskiej. (in:) e . j e n d r z e j c z a k (ed.). przyroda polski w europejskim dziedzictwie dóbr natury. materiały 53 zjazdu ptb toruń bydgoszcz: 133 134. f r i e d r i c h s . , o r z e c h o w s k a m . 2002. macromycetes w środowisku miejskim szczecina. bad. fizjogr. nad pol. zach. ser. b. 51: 7 30. h a n s e n l., k n u d s e n h. 1992 (eds). nordic macromycetes. 2. polyporales, boletales, agaricales, russulales. nordsvamp, copenhagen. h a n s e n l., k n u d s e n h. 1997. (eds). nordic macromycetes. 3. heterobasidioid, aphyllophoroid and gasteromycetoid. basidiomycetes. nordsvamp, copenhagen. h a n s e n l., k n u d s e n h. 2000. (eds). nordic macromycetes. 1. ascomycetes. nordsvamp, copenha gen. k i r k m . p. , c a n n o n p. f. , d a v i d j . c., s t a l p e r s j . a . 2001. ainsworth & bisby’s dictionary of the fungi. ed. 9th. cab international, wallingford. k o n d r a c k i j . 1998. geografia regionalna polski. pwn, warszawa. k r e i s e l h. 1987. pilzflora der deutschen demokratischen republik. basidiomycetes (gallert , hut und bauchpilze). veb gustav fischer verlag jena. k r i e g l s t e i n e r g . j . 1991. verbreitungsatlas der groβpilze deutschlands (west). band 1: ständer pilze. teil a: nichtblätterpilze, teil b: blätterpilze. verlag eugen ulmer, stuttgart. k r i e g l s t e i n e r l . 1999. pilze im naturraum mainfränkische platten und ihre einbindung in die ve getation. regensb. mycol. schr. 9: 1 906. m a t u s z k i e w i c z w. 2001. przewodnik do oznaczania zbiorowisk roślinnych polski. pwn, warszawa. m i r e k z . , p i ę k o ś m i r k o w a h . , z a j ą c a . , z a j ą c m . 2002. flowering plants and pterido phytes of poland a checklist. (in:) z. m i r e k (ed.). biodiversity of poland. 1. w. szafer institute of botany, polish academy of sciences, kraków. m o s e r m . 1949. untersuchungen über den einfluss von waldbränden auf die pilzvegetation i. sydo wia, ann. mycol. 3 (1/6): 336 383. w o j e w o d a w., ł a w r y n o w i c z m. 1992. red list of threatened macrofungi in poland. (in:) k. z a r z y c k i , w. w o j e w o d a , z. h e i n r i c h (eds). list of threatened plants in poland. 2 ed.: 27 56. w. szafer institute of botany, polish academy of sciences, kraków. w o j e w o d a w. 2003. checklist of polish larger basidiomycetes. (in:) z. m i r e k (ed.). biodiversity of poland. 7. w. szafer institute of botany, polish academy of sciences, kraków. z a r ę b a r. 1986. puszcze, bory i lasy polski. pwril, warszawa. zagrożone i chronione macromycetes puszczy wkrzańskiej s t r e s z c z e n i e w pracy przedstawiono charakterystykę występowania wybranych taksonów grzybów wiel koowocnikowych puszczy wkrzańskiej. jest to część wyników systematycznych badań miko socjologicznych prowadzonych na 21 stałych i 64 sporadycznych powierzchniach obserwacyj nych, w latach 2002 2005. w wyniku tych badań stwierdzono 460 gatunków macromycetes, spośród których w niniejszej pracy zaprezentowano 86 gatunków. są to taksony znajdujące się na czerwonej liście grzybów zagrożonych w polsce (84 gatunki) oraz grzyby chronione w polsce (11 gatunków). charakterystyka poszczególnych gatunków obejmuje: kategorię za grożenia i formę ochrony, podłoże, na którym rozwijały się owocniki, zbiorowisko roślinne, ilościowość wytwarzania owocników, okres występowania owocników oraz liczbę i wykaz sta nowisk. 2014-01-01t11:44:24+0100 polish botanical society 2014-09-06t20:23:21+0200 polish botanical society 2014-11-20t23:12:29+0000 polish botanical society 2014-11-20t23:19:15+0000 polish botanical society 2014-11-20t23:14:13+0000 polish botanical society 2014-11-20t23:21:33+0000 polish botanical society 2014-11-20t23:13:01+0000 polish botanical society 2014-11-20t23:13:47+0000 polish botanical society 2014-11-20t23:11:57+0000 polish botanical society 2014-11-20t23:12:21+0000 polish botanical society 2014-09-06t20:23:05+0200 polish botanical society 2014-11-20t23:15:34+0000 polish botanical society 2014-11-20t23:20:18+0000 polish botanical society 2014-11-20t23:16:09+0000 polish botanical society 2014-11-20t23:13:58+0000 polish botanical society 2014-11-20t23:19:53+0000 polish botanical society 2014-11-20t23:22:08+0000 polish botanical society 2014-11-20t23:10:40+0000 polish botanical society 2014-11-20t23:11:21+0000 polish botanical society 2014-11-20t23:11:43+0000 polish botanical society 2014-09-06t20:22:49+0200 polish botanical society 2014-08-20t21:44:53+0200 polish botanical society 2014-11-20t23:14:49+0000 polish botanical society 2014-11-20t23:16:23+0000 polish botanical society 2014-11-20t23:14:37+0000 polish botanical society 2014-11-20t23:20:58+0000 polish botanical society 2014-11-20t23:18:31+0000 polish botanical society 2014-11-20t23:17:23+0000 polish botanical society 2014-09-06t20:23:30+0200 polish botanical society 2014-11-20t23:19:18+0000 polish botanical society 2014-11-20t23:12:32+0000 polish botanical society 2014-11-20t23:14:10+0000 polish botanical society 2014-11-20t23:21:36+0000 polish botanical society 2014-11-20t23:12:05+0000 polish botanical society tubakia dryina, symptoms and pathogenicity to quercus robur tadeusz kowalski department of forest pathology, university of agriculture al. 29 listopada 46, pl 31 425 kraków rltkowal@cyf kr.edu.pl k o w a l s k i t.: tubakia dryina, symptoms and pathogenicity to quercus robur. acta mycol. 41 (2): 299 304, 2006. in 1999 disease symptoms on leaves of quercus robur necrosis, deformation and blackening of leaf petiole, followed by premature leaf fall were observed. tubakia dryina was isolated from necrotic tissues and its pathogenicity to oak proved in infection experiments. the fungus caused necrosis of shoots and leaves. dying leaves displayed also blackening of leaf petiole. key words: quercus, tubakia dryina, pathogenicity, disease symptoms introduction infection of quercus robur l. leaves by several fungi results in easily recognizable disease symptoms. microsphaera alphitoides griff. et maubl. causes white powdery mildew on leaf surface (m a ń k a 2005), taphrina caerulescens (desm. et mont.) tul. – discoloured spots, raised above and conclave below the blade (e l l i s , e l l i s 1985). apiognomonia quercina (kleb.) hoehn. is causative agent of anthracnose, with blade necroses developed along veins (n e l l y , h i m e l i c k 1967). there are, however, numerous diseases causing spots on leaves. their causing agents are difficult to identify due to lack of etiological symptoms, as it is in the case of septoria quercicola (desm.) sacc., cryptocline cinerescens (bubak) arx or mycosphaerella spp. (g i l m a n , wa d l e y 1952; b u t i n 1996). some symptoms, in particular, are not specific as they occur on leaves previously damaged by gall-making insects (b u t i n 1992). various symptoms of unknown etiology are common on oak leaves. the conditions favouring their occurrence are not known yet (b u t i n 1996; k o w a l s k i , d u r a k 2000). a few years ago not specific symptoms of this kind were observed on q. robur leaves, leading to premature defoliation in spring. the symptoms are characterized in the work and results of investigation on the disease etiology are presented. acta mycologica vol. 41 (2): 299-304 2006 dedicated to professor alina skirgiełło on the occasion of her ninety fifth birthday 300 t. kowalski materials and methods leaves of q. robur with disease symptoms were collected in june 1999 near opole (south-west poland). the symptoms were described in detail and isolations of fungi from necrotic tissues were performed. leaf disinfection followed the method used for beech leaves by s i e b e r and h u g e n t o b l e r (1987). leaf bases with petioles were dipped in 96% ethanol (1 min), then in sodium hypochlorite containing ca 4% of active chlorine (3 min) and again in 96% ethanol (0.5 min). after drying in sterile filter paper, from leaf petioles with symptoms 3-mm-long inocula were taken and placed onto 2% malt extract agar (mea, difco) in petri dishes. incubation for 4 weeks was performed in dark, at room temperature. the isolation from necrotic tissues yielded almost exclusively tubakia dryina. four cultures of the species were grown on mea and potato dextrose agar (pda, difco) at 20o c, to describe their morphology and to examine and measure their conidiospores (macroand microconidia). infection experiment on leaves was performed on q. robur trees (ca 10-year-old) in the vicinity of the ojców national park of nature. isolate no. hmipc 16 617 was used for inoculation. developing oak leaves (n 20) were covered on may 2, 2005, and may 19, 2005, with spore suspension (ca 102 viable conidia per ml) obtained from 6-8-week-old cultures grown on mea. besides, at the same time, young shoots (n 6) were wounded with sterile lancet. in the wounds (3-mm-long) going along the shoot an inoculum of ca 5mm2 of mea grown with t. dryina mycelium was placed and the shoot covered with parafilm. control consisted of wounded shoots with sterile medium. four and eight weeks after inoculation the disease symptoms were evaluated and the fungi isolated from 210 fragments of necrotic leaf tissue, leaf petioles and shoots, as described above. results the disease symptoms on q. robur leaves observed in 1999 near opole, were mainly visible on leaf petioles. necrosis, blackening and deformation by bending down or twisting were extremely characteristic. in some cases the necrosis spread to blade base (fig. 1). such leaves fell down prematurely, in spring. from dead leaf petioles almost entirely one fungal species was isolated – tubakia dryina. sporadically also fungi representing the genera of alternaria, coniothyrium and epicoccum were isolated. tubakia dryina colony on mea was woolly to floccose, reaching after 21 days diameter of 86 mm (on average), whitish at first, then olive-grey (fig. 2). on pda the growth was slower (72 mm after 21 days, on average), the structure was more compact, with concentric rings more distinct cloudy, with deep bay-like cuts at the circumference (fig. 3). black sporodochia formed concentric rings on the colony surface and produced numerous conidia of two types. macroconidia were 1-celled, hyaline to light olive-brown, elliptical, sometimes truncate at the basis, 12-15 x 7-8 μm (fig. 4). beside macroconidia also 1-celled hyaline, rod-shaped to oblong elliptical microconidia measuring 6-8 x 2 μm (fig. 4) were formed. over a half of artificially inoculated leaves (65.0%) and all inoculated shoots displayed disease symptoms after 4-8 weeks. tissue along vein in the upper part of tubakia dryina 301 leaves was necrotic (fig. 5), and some leaves died (fig. 6). wounds in the inoculated shoots did not heal, and the tissues around – particularly above wounds, were necrotic and discoloured brown-black (fig. 7). in the leaves occurring there, leaf petiole was necrotized and black which resulted in dieback of entire leaves (figs 7, 8). the necrotized leaves were not properly developed and displayed atrophy (fig. 6). in the necrotic tissue t. dryina only sporadically produced conidiomata of pycnothyrium type. out of 210 samples of tissues from inoculated leaves and shoots which were necrotized, 59.6% yielded microorganisms. tubakia dryina was found in all plant parts in question (leaves, leaf petioles, shoots) and isolated from 28.6% of tissue samples. the following species were isolated beside t. dryina: alternaria alternata (fr.) keissler (0.5%), apiognomonia quercina (5.7%), aureobasidium pullulans (de bary) arn. (3.8%), coniothyrium olivaceum sacc. (2.4%), fusarium sp. (10.0%), hormonema sp. (1.0%) and lecytophora sp. (0.5%). from 7.1% of tissue samples bacterial colonies were obtained, which displayed in vitro antagonistic effect on t. dryina colony. after 2 weeks of common growth (in two organism cultures) on mea (inoculum distance 25 mm), an inhibition zone was produced of 3-15 mm, and the t. dryina colony radius towards the bacterial colony was reduced 3-5 times (fig. 9). discussion tubakia dryina [syn. actinopelte dryina (sacc.) hoehn.] occurs in north america, europe and asia, while its teleomorph, described as dicarpella dryina belisario, was described only in italy (h o l d e n r i e d e r , k o w a l s k i 1989; p r o f f e r 1990; b e l i s a r i o 1991; b u t i n 1996; k a n e k o , k a n e k o 2004). it is found mainly on oaks, and rarely on other plants of acer, castanea, liquidambar, photinia, sasafras and ulmus genera (p r o f f e r 1990). it was described in poland only once – by p r z y b y ł (1995). in plants the species is able to occur as endophyte, causing no disease symptoms (b o d y , r a y n e r 1984; p r z y b y ł 1995; c o h e n 1999; g e n n a r o et al. 2003; k a n e k o , k a n e k o 2004). it is also being connected with disease symptoms on leaves and, rather seldom, on shoots (h o l d e n r i e d e r , k o w a l s k i 1989; b u t i n 1996). on quercus spp. leaves the symptoms are circular, tan to dark reddish brown spots, 1-15 mm in diameter. small spots may coalesce to form large irregular blotches (h e p t i n g 1971; m u n k v o l d , n e e l y 1990; p r o f f e r 1990). on castanea leaves brown necrotic spots reach up to 9 mm in diameter and the lesions are surrounded by a prominent chlorotic halo, up to 6 mm wide (e l g h o l l et al. 1996). pathogenicity tests with t. dryina yielded inconsistent results. artificial inoculation of q. phillyraeoides, q. glauca and castanea pubinervis performed by yo k o y a m a and tu b a k i (1971) revealed no pathogenicity. the t. dryina isolate examined by h o l d e n r i e d e r and k o w a l s k i (1989) did not display any pathogenicity towards shoots and very low pathogenic potential on leaves of q. robur. on the other hand, e l g h o l l et al. (1996), k i m and wa g n e r (1997), and m u n k v o l d and n e l l y (1990) confirmed pathogenicity of the species. the t. dryina isolate tested by the author proved also pathogenic. despite the fact that it was not morphologically different from the isolates investigated by other authors (yo k o y a m a , tu b a k i 1971; h o l d e n r i e d e r , k o w a l s k i 1989; p r o f f e r 1990), it caused different type of symptoms on leaves – a very specific necrosis and blackening of leaf petioles, and necrosis of leaf blade 302 t. kowalski – at first along veins and than of the entire leaf. the necrosis spread so quickly that the leaves did not grow to normal size. the symptoms, very different from those observed by other authors (m u n k v o l d , n e l l y 1990; p r o f f e r 1990; e l g h o l l et al. 1996), could have resulted from infection of very young developing leaves of q. robur. that early artificial inoculation was aimed at finding out if t. dryina could cause necrosis together with malformation and blackening of leaf petioles, leading – according to the 1999 observations – to premature leaf dropping. the results confirmed the hypothesis, which is of interest as the symptoms described were never observed in connection with t. dryina infection. they also pointed to the possible effect of bacteria inhabiting q. robur leaves on development of t. dryina. acknowledgements: the author thanks mrs mgr j. michalik for help in laboratory work. these investiga tions were carried out under the project no.2 p06l 036 26. references b e l i s a r i o a. 1991. dicarpella dryina sp.nov., teleomorph of tubakia dryina. mycotaxon 41 (1): 147 155. b o d d y l., r a y n e r a. d. m. 1984. fungi inhabiting oak twigs before and at fall. transactions of the british mycological society 82: 501 505. b u t i n h. 1992. effect of endophytic fungi from oak (quercus robur l.) on mortality of leaf inhabiting gall insects. eur. j. for. path. 22: 237 246. b u t i n h . 1996. . krankheiten der wald und parkbaeume . georg thieme verlag. stuttgart. c o h e n s. d. 1999. technique for large scale isolation of discula umbrinella and other foliar endophytic fungi from quercus species. mycologia 91 (5): 917 922. e l g h o l l n. e., s c h u b e r t t. s., p e a c o c k m. e. 1996. tubakia leaf spot of chestnut. plant pathol ogy circular. 375: 1 3. e l l i s m. b., e l l i s j. p. 1985. microfungi on land plants. london, sydney: croom helm. g e n n a r o m . , g o n t h i e r p. , n i c o l o t t i g . , 2003. fungal endophytic communities in healthy and declining quercus robur l. and q. cerris l. trees in northern italy. j. phytopathol. 151: 529 534. g i l m a n j. c., wa d l e y b. n., 1952, the ascigerous stage of septoria querceti thuem. mycologia 44: 216 220. h e p t i n g g. h. 1971. diseases of forest and shade trees of the united states. usda forest service. agricultural handbook no. 386. 658 pp. h o l d e n r i e d e r o . , k o w a l s k i t. 1989. pycnidial formation and pathogenicity in tubakia dryina. mycol. res. 92 (2): 166 169. k a n e k o r., k a n e k o s. 2004. the effect of bagging branches on levels of endophytic fungal infection in japanese beech leaves. for. path. 34: 65 78. k i m s. h., wa g n e r v. m. 1977. parasitism of actinopelte dryina on oak leaves (abstr.) proceedings of american phytopathological society 4: 125 126. k o w a l s k i t., d u r a k e. 2000. grzyby endofityczne iv. mykobiota w żywych liściach quercus robur oraz w nekrotycznych tkankach wokół wyrośli andricus ostrea. zesz. nauk. ar kraków 376: 25 36. m a ń k a k . 2005. fitopatologia leśna. pwril. warszawa. m u n k v o l d g. p., n e e l e y d. 1990. pathogenicity of tubakia dryina. plant disease 74: 518 522. n e e l y d., h i m e l i c k e. b. 1967. characteristick and nomenclature of the oak antracnose fungus. phytopathology 57: 1230 1236. p r o f f e r t. j. 1990. tubakia leaf spot. florida department of agriculture and consumer services, divi sion of plant industry, gainesville. plant pathology circular no. 337. 2 p. p r z y b y ł k. 1995. zamieranie dębów w polsce. idee ekologiczne. wyd. sorus, poznań, 8 (4): 1 85. s i e b e r t., h u g e n t o b l e r c. 1987. endophytische pilze in blättern und ästen gesunder und ge schädigter buchen (fagus sylvatica l.). eur. j. for. path. 17: 411 425. yo k o y a m a t., tu b a k i k. 1971. cultural and taxonomical studies on the genus actinopelte. research communications, institute for fermentation, osaka 5: 43 77. tubakia dryina 303 tubakia dryina – patogeniczność i symptomy na quercus robur s t r e s z c z e n i e w pracy przedstawiono wyniki badań nad objawami chorobowymi obserwowanymi w 1999 r. w rejonie opola u quercus robur w postaci nekrozy, deformacji i czernienia ogon ka liściowego, co prowadziło do przedwczesnego opadu liści. z martwych tkanek izolowano prawie wyłącznie kolonie jednego gatunku grzyba, który zidentyfikowano jako tubakia dry ina. gatunek ten wytwarzał in vitro zarówno makro jak i mikrokonidia, wykazując nieco szybszy wzrost na pożywce agarowomaltozowej niż na pożywce glukozowoziemniaczanej. po przez sztuczną inokulację liści i pędów q. robur wykazano patogeniczne właściwości bada nego szczepu t. dryina. scharakteryzowano różne typy objawów chorobowych uzyskanych w trakcie tego eksperymentu. zwrócono uwagę na ich odmienność w porównaniu z objawami obserwowanymi w innych krajach. 2014-01-01t11:44:42+0100 polish botanical society 2014-11-20t23:12:00+0000 polish botanical society 2014-11-20t23:13:36+0000 polish botanical society 2014-11-20t23:18:00+0000 polish botanical society 2014-09-06t20:23:09+0200 polish botanical society 2014-11-20t23:15:26+0000 polish botanical society 2014-11-20t23:20:10+0000 polish botanical society 2014-11-20t23:14:01+0000 polish botanical society 2014-11-20t23:16:03+0000 polish botanical society 2014-11-20t23:20:35+0000 polish botanical society 2014-11-20t23:21:50+0000 polish botanical society © the author(s) 2014 published by polish botanical society microfungi of the tatra mts. 6. fungus-like organisms: albuginales, peronosporales and pythiales wiesław mułenko1, monika kozłowska1*, kamila bacigálová2, urszula świderska-burek1 and agata wołczańska1 1department of botany and mycology, institute of biology and biochemistry, maria curie-skłodowska university, akademicka 19, pl-20-033 lublin, *corresponding author: monika@poczta.umcs.lublin.pl 2institute of botany, slovak academy of sciences, dúbravská cesta 9, sl-845 23 bratislava 4 mułenko w., kozłowska m., bacigálová k., świderska-burek u., wołczańska a.: microfungi of the tatra mts. 6. fungus-like organisms: albuginales, peronosporales and pythiales. acta mycol. 49 (1): 3–21, 2014. a list and the distribution of oomycota species in the tatra mts (western carpathian mts) are presented. revised herbarium vouchers and literature data were used for analysis. thirty two species of oomycetes on fifty seven plant species were noted in the area, including two species of the order albuginales (genera: albugo and pustula, on nine plant species), 29 species of the order peronosporales (genera: bremia, hyaloperonospora, peronospora and plasmopara, on 49 plant species), and one species of the order pythiales (genus: myzocytium, on one species of algae). twenty nine species were collected on the polish side of the tatra mts and ten species were collected on the slovak side. the oomycetes were collected at 185 localities. key words: microfungi, distribution, oomycota, western carpathians, poland, slovakia introduction the phylum oomycota is a small group of approximately 1 000 species (kirk et al. 2008), including aquatic and terrestrial, parasitic and saprobic fungus-like organisms which cause numerous plant diseases. peronosporales, which mainly consists of obligatory parasites of vascular plants, is the largest order in the phylum. three families were previously distinguished in this order: the albuginaceae, peronosporaceae and pythiaceae (kochman, majewski 1970). however, modern taxonomy classifies them as three separate orders: albuginales, peronosporales and pythiales (kirk et al. 2008). species of these orders that are economically important as causative agents of considerable losses in cultivated crops are well recognized and described. many studies on these organisms are important taxonomic monographs comprising the acta mycologica vol. 49 (1): 3–21 2014 doi: 10.5586/am.2014.001 dedicated to professor maria ławrynowicz on the occasion of the 45th anniversary of her scientific activity 4 w. mułenko et al. © the author(s) 2014 published by polish botanical society entire order peronosporales (in the former classification) (kochman, majewski 1974) or smaller taxonomic units [e.g. plaats-niterink (1981); saharan et al. (1997)]. much less is known about the diversity and abundance of these species in natural conditions, on plants growing in the wild. only a few systematic studies of this group are identified in the polish mycosociological literature spanning a period of over 50 years of research (ławrynowicz et al. 2004). downy mildews in alpine conditions were either not included in these studies or were investigated only sporadically. the massif of the tatra mts is one of the most interesting alpine regions in central europe and is almost entirely contained within national parks in poland (tpn, tatrzański park narodowy) and in slovakia (tanap, tatranský národný park). it was declared a unesco biosphere reserve in 1992. while floristic, ecological, geological, hydrological and other research has been successfully conducted in the tatra mts (vološčuk 1994; mirek et al. 1996), detailed mycological investigations have not been carried out in the region. the geobotanical differentiation and the richness of plant species that are potential hosts of fungi in the region are high, and it can be expected that many interesting species of fungi and oomycetes, can occur in the area. results of studies to date indicate that microfungi in the tatra mts are poorly recognized. in-depth information on their ecology and distribution is not available and data are sometimes random. consequently, information concerning the actual spread of fungi is highly differentiated and the data from individual areas of the tatra mts are considerably deficient. the majority of studies published so far are simple species lists or brief notes on a particularly interesting or rare species. due to the lack of easy access to the area, analytical studies are undertaken infrequently (e.g. urban 1952; bacigálová et al. 2005; mułenko et al. 2006, 2008). a similar problem is observed in other mountainous regions in europe and worldwide. the aim of our study was to summarize the existing literature data on the oomycota occurring throughout the tatra massif within the two national parks. more extensive research can be based on our results in the future. a similar study was carried out for representatives of the order taphrinales in the mid-2000s (bacigálová et al. 2005). the present paper is part of the series named parasitic microfungi of the tatra mts. five studies have been published to date: representatives of the order taphrinales (bacigálová et al. 2005), pseudocercosporella tetrensis (mułenko, bacigálová 2005), spermosporina gymnadeniae (wołczańska et al. 2008), melampsoridium hiratsukanum (mułenko et al. 2005) and the plasmopara spp. on the members of the genus geranium (mułenko et al. 2008). as only biotrophic species infect living plant organs and a considerable part of the collection are fungi occurring on decaying or dead organs (facultative parasites, facultative saprophytes), the word “parasitic” will now be omitted in the series title. although this paper does not concern true fungi we left the name “microfungi” in the title of the series, because oomycetes are only a small part of all microorganisms which are published in this series. microfungi of the tatra mts. 6. 5 © the author(s) 2014 published by polish botanical society history of the research: an outline research into microfungi in the tatra mts goes back to the end of the 19th century when hazslinszky (1864) published a list of fungi collected in this area. the first information on downy mildews can be found in a study by krupa (1886) and refers to albugo candida on arabis alpina on the polish side of the tatra mts. in 1888 krupa expanded the list of hosts by further four species: arabis soyeri subsp. subcoriacea, cardaminopsis halleri, hutchinsia alpina and kernera saxatilis. data on oomycetes were published at the beginning of the 20th century by rouppert (1912), wróblewski (1918, 1920, 1922a, 1922b, 1925) and szulczewski (1930). the above-mentioned works included lists of species occurring on the polish side of tatra mts. however, wróblewski collected fungi on both sides of the border and was the first to publish information on microfungi in the slovak part of the tatra mts. further research in this region was carried out by moesz (1930) and picbauer (1933). mainly the oomycota occurring on the polish side of the tatra mts were discussed in the literature in the second half of the 20th century: starmachowa (1963), kućmierz (1968), kochman & majewski (1970), sałata et al. (1984), romaszewskasałata et al. (1986), and mułenko et al. (1995). a species parasitizing algae, myzocytium megastomum on closterium lunula, was reported in 1999 (kadłubowska 1999). only two studies investigating the slovak side of the tatra mts were published in that period (müller 1980, skalický 1983). investigations in the tatra national park (tpn, tanap) intensified in the late 20th and early 21st centuries. results of studies conducted jointly with the slovak academy of science (sav) in the entire region of the tatra mts were first reported. for example, a detailed analysis of plasmopara spp. on the members of geranium was conducted by mułenko et al. (2008). specimens from the tatra mts were also examined during research into plasmopara praetermissa, a species new to science (voglmayr et al. 2006). both original results and summative studies have been reported in the literature. the first list of fungi of the tatra mts dates from the early 20th century and can be found in a study on the fungi and slime moulds of two historical regions: galicia and bukovina (namysłowski 1914). however, a monograph by starmachowa (1963) is the first synthesis of microfungi. it comprises the entire tatra massif, i.e. both the polish and the slovak sides. the monograph was the main source of microfungirelated information for over thirty years until the late 20th century. an extensive discussion of the results of mycological research conducted in the polish tatra mts was given in a monograph on the tatra national park (tpn, pl) (sałata, mułenko 1996). however, a list of species was not included due to both popular and scientific focus of the study. a full list of fungi and fungus-like organisms, considerably extended in comparison with the study by starmachowa (1963), was published in 2004 (mułenko et al.). a similar study for the slovak part of the national park was completed by bacigálová (1999). 6 w. mułenko et al. © the author(s) 2014 published by polish botanical society material and methods the list of species was based on literature reports published in the last 128 years. the oldest publications did not always contain detailed data. some records were given in very general terms and only information that a species had been found in the tatra mts was reported while the location, site or study area were not indicated and the polish or slovakian part of the region was not specified. some of the specimens listed in this work have not survived. the majority, however, are deposited in the herbaria in poland (kra and kram in kraków, wauf in warsaw, lbl in lublin) and in slovakia (sav in bratislava) (acronyms according to holmgren et al. 1990). in some cases information from exsiccatum labels was used, which was not published previously. wherever possible, we provided the habitat of the oomycetes listed. results since 1886 a total of 32 species belonging to the oomycota have been collected in the tatra national park. they are representatives of three orders: albuginales (two species), peronosporales (29 species) and pythiales (one species). the fungus-like organisms were collected on 56 species of vascular plants and on one species of algae (tab. 1). the number of reported species is not very high and oomycetes are underexplored in the tatra mts. mycosociological research in poland to date shows that the abundance of the species of the oomycota is greater, not only in considerably smaller regions but also in less floristically diversified areas. majewski (1967) collected 58 species in the puszcza kampinoska forest, kućmierz (1973) collected 59 species in table 1 general distribution of the oomycete in the tatra mts according to number of localities for each species species host plant slovakia number of localities poland number of localities total number zt vt bt wl tz tw wl albuginales albugo candida arabis alpina 1 2 7 10 arabis soyeri subsp. subcoriacea 1 1 biscutella laevigata 1 3 4 capsella bursa-pastoris 1 1 cardaminopsis arenosa 1 3 4 cardaminopsis halleri 3 3 hutchinsia alpina 2 2 kernera saxatilis 1 1 pustula tragopogonis cirsium palustre 1 1 1 4 21 1 27 microfungi of the tatra mts. 6. 7 © the author(s) 2014 published by polish botanical society peronosporales bremia lactucae carlina acaulis 3 3 cicerbita alpina 1 1 cirsium erisithales 1 1 crepis paludosa 1 1 hieracium aurantiacum 1 1 hieracium murorum 1 1 hieracium vulgatum 1 1 6 8 senecio subalpinus 5 5 senecio vulgaris 1 1 taraxacum officinale 1 1 hyaloperonospora arabidisalpinae arabis alpina 1 1 hyaloperonospora cardaminopsidis cardaminopsis arenosa 1 1 hyaloperonospora lunariae lunaria rediviva 3 3 hyaloperonospora parasitica cardamine amara 1 1 cardamine trifolia 1 1 peronospora alpicola ranunculus platanifolius 3 3 peronospora alta plantago major 7 7 peronospora boni-henrici chenopodium bonushenricus 3 3 peronospora conferta cerastium fontanum 2 2 cerastium holosteoides 2 2 cerastium tatrae 1 1 peronospora debaryi urtica urens 1 1 peronospora erysimi erysimum hieraciifolium 1 1 peronospora galii galium anisophyllon 1 1 galium schultesii 3 3 peronospora grisea veronica beccabunga 1 1 veronica serpyllifolia 1 1 peronospora knautiae scabiosa lucida 3 3 peronospora mayorii vicia cracca 1 1 peronospora myosotidis myosotis palustris 1 1 peronospora phyteumatis phyteuma spicatum 2 2 peronospora potentillae potentilla aurea 2 8 10 peronospora ranunculi ranunculus oreophilus 1 1 ranunculus repens 3 3 peronospora rumicis rumex acetosa 2 2 rumex acetosella 1 1 rumex alpestris 1 1 peronospora septentrionalis cerastium cerastoides 1 1 peronospora sordida scrophularia scopolii 3 3 peronospora trifoliorum lathyrus pratensis 1 1 trifolium repens 1 1 peronospora violae viola biflora 1 1 plasmopara densa rhinanthus serotinus 1 1 plasmopara geranii-silvatici geranium palustre 1 1 geranium pratense 1 1 2 geranium sylvaticum 11 6 5 19 41 geranium sp. 1 1 2 table 1 – cont. 8 w. mułenko et al. © the author(s) 2014 published by polish botanical society the ojców national park and 73 species in the pieniny mts (kućmierz 1977). similar investigations were carried out in the bug river valley and in the polesie national park, from which 85 species (danilkiewicz 1987) and 55 species (mułenko 1988 a, b) were reported respectively. in other areas, however, a smaller number of oomycetes was noted. for instance, 36 species were recorded in the białowieża national park (majewski 1971), 31 species in the słowiński national park (adamska 2001), and 36 species in the jura krakowsko-częstochowska upland (ruszkiewicz-michalska 2006). as an analysis of the list of downy mildews in the tatra mts shows, the current state of knowledge of them varies considerably in poland and slovakia. the majority of species were reported from the polish side of the tatra mts (29) while only ten were noted on the slovak side (tab. 2). the numbers of species found in individual parts of the tatra mts are highly divergent. the greatest numbers of fungus-like organisms were collected in the western tatras – 26 species. all of them (26 species) were recorded on the polish side and only two on the slovak side. thirteen species were reported in the high tatra mts, including nine in slovakia and six in poland. two of them (plasmopara nivea and plasmopara pratermissa) are common to both countries. four species were recorded in the belianské tatry mts, including one (peronospora septentrionalis) that is known only from this region. the spread of oomycetes, as measured by the number of existing localities, varies considerably. in total, fungus-like organisms were recorded at 185 localities. as plasmopara nivea aegopodium podagraria 2 2 anthriscus sylvestris 1 1 mutellina purpurea 1 1 2 plasmopara praetermissa geranium sylvaticum 2 2 2 1 7 geranium sp. 1 1 plasmopara pusilla geranium pratense 5 1 6 geranium sylvaticum 1 2 3 14 20 10 1 104 8 157 pythiales myzocytium megastomum closterium lunula 1 1 1 1 number of species number of host plants total number of localities 32 57 14 21 14 1 126 8 1 185 denotations: zt – západné tatry mts (sk); vt – vysoké tatry mts (sk); bt – belianské tatry mts (sk); tz – tatry zachodnie mts (pl); tw – tatry wysokie mts (pl); wl – without precise localization table 2 number of oomycete species in particular regions of tatra mts (denotations see table 1) slovakia poland zt vt bt wl tz tw wl 2 9 4 1 26 6 1 total number of species: 10 total number of species: 29 table 1 – cont. microfungi of the tatra mts. 6. 9 © the author(s) 2014 published by polish botanical society many as 135 records (73%) were noted on the polish side of the tatra mts and 126 (68%) were found in the western tatra mts. oomycetes were noted at 50 localities on the slovak side (27%), mainly in the vysoké tatry mts (21 records). the number of localities is the same in the remaining regions (západné tatry mts and belianské tatry mts; 14 in each) (tab. 1). polyphagous species were recorded more frequently. they occurred sporadically on individual hosts. the most frequent combinations were: albugo candida/arabis alpina and peronospora potentillae/potentilla aurea (10 records each), bremia lactucae/hieracium vulgatum (8 records) and peronospora alta/plantago major (7 records). of the above-mentioned species, only species of the genus plasmopara occurring on geranium spp. have been examined in greater depth in the last few years and the research data have been published (mułenko et al. 2008). the species were found at 63 localities: 26 in the polish and 37 in the slovak part of the tatra mts. the study provides the only relatively objective results on the research into the spread of fungus-like organisms in the region available to date. there is a great need for further intensive and systematic research in the area. list of oomycota arrangement: the list of species is organized alphabetically by order name (albuginales, peronosporales, pythiales) and by species name within the orders. host plant species are listed alphabetically. names of species of oomycetes are in bold. names of doubtful species are not given in bold (see below). notes: explanations are placed in two ways. if a note refers to a particular plant species (or an oomycete on this plant), it is placed at the end of the paragraph relating to the plant. if a note refers to a particular oomycete species (regardless of the host), it is placed as an independent paragraph below the list of host plants. doubtful species: some species of oomycetes and of plants may be doubtful or may have been reported incorrectly. to retain the information given in the literature, names of such species are given in full but a note or a reference is added. neither the name of an oomycete species nor the name of the plant is given in bold in such instances. names of oomycetes: names of oomycetes are given after kochman & majewski (1970), constantinescu (1991), thines & spring (2005), and index fungorum (http://www.indexfungorum.org/). old names of oomycetes are given for some species, that is the names under which they were previously published, while some of them are not synonymous. only the names cited in studies regarding this region are given in the synonyms. names of plants: names of host plants are given after mirek et al. (2002) and marhold & hindak (1998). in some plant species their synonyms or their old names, under which they were published previously, are given. only the names cited in studies referring to the tatra mts were used. division of the tatra mts: the entire tatra mts system is divided into regions differentiated on both sides of the tatras: western tatra mts – tatry zachodnie (pl) and západné tatry mts (sk). eastern tatra mts – tatry wysokie (pl), vysoké tatry mts (sk) and belianské tatry mts (sk). the respective regions of the tatra mts follow local names used on tatra mts tourist maps no. 112 and no. 113 (1:50.000), published by vojensky kartograficky ustav (harmanec, slovakia, 1992-1996) and “wielka encyklopedia tatrzańska” (radwańska-paryska & paryski 2004). abbreviations: tnp – tatra national park; pl – polish side of the tatra mts; sk – slovak side of the tatra mts; v-x – collection months. literature sources and citations: not all the authors mentioned in the section “history of the research: an outline” provided data on their own collections. many authors cited previously published information. this method is a common practice, particularly in the preparation of taxonomic or synthetic studies. however, the list of authors may be wrongly identified with a number of known locations in this way. thus the original data source is given for each species in this study, whereas citations are provided separately and placed as supplementary information below the description of the locality. 10 w. mułenko et al. © the author(s) 2014 published by polish botanical society oomycota albuginales albugo candida (pers.) roussel [cystopus candidus (pers.) lév.] – arabis alpina l. tnp: pl – tatry zachodnie mts: dolina strążyska valley; viiviii (krupa 1886, 1888; rouppert 1912; starmachowa 1963). dolina kościeliska valley; vii-ix (krupa 1888; wróblewski 1918, 1922b). dolina mała łąka valley; viii-ix (krupa 1888; rouppert 1912). dolina białego valley; mała dolinka pod giewontem valley; vii-viii (wróblewski 1922b). przełęcz iwaniacka pass; vii (kućmierz 1968). cited by namysłowski (1914), wróblewski (1920, 1925), starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). sk – vysoké tatry mts: belanská kopa mt. at alt. 1690 m (moesz 1930). belianské tatry mts: kopské sedlo pass; tatranská javorina village (picbauer 1933). cited by starmachowa (1963) and bacigálová (1999). – arabis soyeri reut. & a. huet subsp. subcoriacea (gren.) breistr. (arabis bellidifolia jacq. non crantz). tnp: pl – tatry zachodnie mts: hruby regiel mt. (krupa 1888). cited by namysłowski (1914), starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). – biscutella laevigata l. tnp: pl – tatry zachodnie mts: dolina strążyska valley; mały giewont mt.; vi-vii (wróblewski 1922b). mały giewont mt. at alt. 1650 m, limestone bedrock; viii (sałata et al. 1984). cited by starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). sk – belianské tatry mts: dolina siedmich prameňov valley (picbauer 1933). cited by starmachowa (1963) and bacigálová (1999). – capsella bursa-pastoris (l.) medik. tnp: pl – tatry zachodnie mts: dolina kościeliska valley; vii, ix (wróblewski 1918). cited by starmachowa (1963) and mułenko et al. (2004). – cardaminopsis arenosa (l.) hayek [arabis arenosa (l.) scop.]. tnp: pl – tatry zachodnie mts: dolina ku dziurze valley (szulczewski 1930). dolina białego valley at alt. 1000 m, calcareous plants; mały giewont mt. at alt. 1650 m, limestone bedrock; viii (sałata et al. 1984). cited by starmachowa (1963) and mułenko et al. (2004). sk – belianské tatry mts: without localization at alt. 1320 m (?bujačí vrch mt.) (moesz 1930). cited by starmachowa (1963) and bacigálová [1999, incorrectly on arabis hirsuta (l.) scop.]. – cardaminopsis halleri (l.) hayek (arabis halleri l.). tnp: pl – tatry zachodnie mts: kuźnice village (krupa 1888). dolina strążyska valley; dolina kościeliska valley; vi, viii (wróblewski 1922b). cited by wróblewski (1920), starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). – hutchinsia alpina (l.) r. br. (lepidium alpinum l.). tnp: pl – tatry zachodnie mts: dolina za bramką valley (krupa 1888). mała dolinka pod giewontem valley; vii (wróblewski 1922b). cited by namysłowski (1914), starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). – kernera saxatilis (l.) rchb. tnp: pl – tatry zachodnie mts: dolina strążyska valley (krupa 1888). cited by namysłowski (1914), starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). pustula tragopogonis (pers.) thines [albugo tragopogonis (dc.) gray, cystopus tragopogonis schröt.] microfungi of the tatra mts. 6. 11 © the author(s) 2014 published by polish botanical society – cirsium palustre (l.) scop. tnp: pl – tatra mts: without localization; viii (wróblewski 1922a, leg. j. krupa, 1886). cited by starmachowa (1963) and kochman & majewski (1970). notes: the herbarium collections need revision because this oomycete was treated as a collective species occurring on representatives of the family asteraceae according to kochman & majewski (1970). this fungus-like organism can be divided into two species: pustula tragopogonis and pustula spinulosa. peronosporales bremia lactucae regel s.l. – carduus personata (l.) jacq. notes: reported by kochman & majewski (1970) from tatra mts (without localization), however the collection was located in zakopane town (wauf, 01 aug 1961, leg. j. kochman). – carlina acaulis l. tnp: pl – tatry zachodnie mts: dolina białego valley; dolina strążyska valley; dolina za bramką valley; vi (wróblewski 1922b). cited by starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). – carlina vulgaris l. notes: starmachowa (1963) and later mułenko et al. (2004) wrongly reported presence of this oomycete at toporowe stawy lakes (pl) after rouppert (1912). however, rouppert (1912) did not mention this species. similarly, bacigálová (1999) reported the oomycete on this plant from vysoké tatry mts (sk) after picbauer (1933), however the latter author did not mention this information. – cicerbita alpina (l.) wallr. tnp: pl – tatry zachodnie mts: ścieżka nad reglami tourist route at alt. 1270 m, herbaceous plants; viii (romaszewska-sałata et al. 1986). cited by mułenko et al. (2004). – cirsium erisithales (jacq.) scop. tnp: pl – tatry zachodnie mts: dolina spa-spadowiec valley at alt. 910 m, herbaceous plants; viii (sałata et al. 1984). cited by mułenko et al. (2004). – crepis paludosa (l.) moench. tnp: pl – tatry zachodnie mts: dolina ku dziu-dziurze valley at alt. 910 m, spruce forest with abies alba; viii (sałata et al. 1984). cited by mułenko et al. (2004). – hieracium aurantiacum l. tnp: pl – tatry zachodnie mts: niżnia świstówka mt. at alt. 1500 m, grassland; ix (mułenko et al. 1995). cited by mułenko et al. (2004). – hieracium murorum l. tnp: pl – tatry zachodnie mts: dolina białego valley at alt. 960 m, dentario glandulosae-fagetum; viii (sałata et al. 1984). cited by mułenko et al. (2004). – hieracium vulgatum fr. tnp: pl – tatry zachodnie mts: dolina olczyska valley at alt. 962 m; dolina ku dziurze valley at alt. 970 m; dolina strążyska valley at alt. 1020 m; boczań mt. at alt. 1150 m; igła mt. at alt. 1150 m; czoła jaworzyńskie hill at alt. 1160 m, spruce forest for all localities; viii (sałata et al. 1984). cited by mułenko et al. 2004. sk – tatra mts: without localization (kochman & majewski 1970). vysoké tatry mts: štrbské pleso lake (picbauer 1933). cited by starmachowa (1963) and bacigálová (1999). notes: the name of hieracium vulgatum fr. has not been changed because according to mirek et 12 w. mułenko et al. © the author(s) 2014 published by polish botanical society al. (2002) it can be a synonym for 2 different species: hieracium lachenalii c.c. gmel and hieracium laevicaule jord. – senecio subalpinus w. d. j. koch. tnp: pl – tatry zachodnie mts: dolina kościeliska valley [wauf, 26 july 1961, leg. t. majewski; reported by kochman & majewski (1970), without localization], dolina strążyska valley at alt. 900 m; dolina ku dziurze valley at alt. 920 m; dolina kościeliska valley (stare kościeliska) at alt. 940 m; igła mt. at alt. 1025 m, herbaceous plants for all localities; viii (sałata et al. 1984). cited by mułenko et al. (2004). – senecio vulgaris l. tnp: pl – tatry zachodnie mts: toporowe stawy lakes; ix (wróblewski 1918). – taraxacum officinale f. h. wigg. tnp: pl – tatry zachodnie mts: dolina kościeliska valley (kram, 10 sept. 1966, leg. t. majewski). reported by kochman & majewski (1970) on taraxacum sp., without localization. cited by mułenko et al. (2004). hyaloperonospora arabidis-alpinae (gäum.) göker, riethm.; voglmayr, weiss & oberw. [peronospora arabidis-alpinae gäum.] – arabis alpina l. tnp: pl – tatry zachodnie mts: ścieżka nad reglami tourist route (near zameczki rocks) at alt. 1200 m, damp alpine grassland (mułenko et al. 1995). cited by sałata & mułenko (1996) and mułenko et al. (2004). hyaloperonospora cardaminopsidis (a. gustavsson) göker, riethm. voglmayr, weiss & oberw. [peronospora cardaminopsidis a. gustavson] – cardaminopsis arenosa (l.) hayek. tnp: pl – tatry zachodnie mts: mały giewont mt. at alt. 1650 m, limestone bedrock; viii (sałata et al. 1984). cited by mułenko et al. (2004. hyaloperonospora lunariae (gäum.) constant. [peronospora lunariae gäum.] – lunaria rediviva l. tnp: pl – tatry zachodnie mts: wąwóz kraków ravine at alt. 1030 m, herbaceous plants on limestone bedrock; viii (sałata et al. 1984) and 1000 m, scrub; vi-viii (mułenko et al. 1995). dolina kościeliska valley at alt. 980 m, dentario glandulosae-fagetum; vi-viii (mułenko et al. 1995). cited by sałata & mułenko (1996) and mułenko et al. (2004). hyaloperonospora parasitica (pers.) constant. [peronospora dentariae rabenh., peronospora parasitica (pers.) fr.] – cardamine amara l. tnp: pl – tatry wysokie mts: włosienica glade at alt. 1320 m, streamside; vi (mułenko et al. 1995). cited by mułenko et al. (2004). – cardamine trifolia l. tnp: pl – tatry zachodnie mts: igła mt. at alt. 1080 m, spruce forest; viii (sałata et al. 1984). cited by sałata & mułenko (1996) and mułenko et al. (2004). notes: the oomycete was also reported by wróblewski (1922b) on hutchinsia alpina (l.) r. br. (lepidium alpinum l.) and later cited by starmachowa (1963). see notes on peronospora lepidii (belove). peronospora alpicola gäum. [peronospora ficariae auct. p.p.] – ranunculus platanifolius l. tnp: pl – tatry wysokie mts: near wodogrzmoty mickiewicza waterfall; vii (wróblewski 1922b). morskie oko lake at alt. 1405 m, herbaceous plants; surroundings of czarny staw gąsienicowy lake at alt. 1610 m, herbaceous plants in pinetum mugo carpaticum; viii (sałata et al. 1984). cited by starmachowa (1963), kochman & majewski (1970), sałata & mułenko (1996) and mułenko et al. (2004). microfungi of the tatra mts. 6. 13 © the author(s) 2014 published by polish botanical society ? peronospora alsinearum casp. [peronospora media gäuman]. notes: the oomycete was reported on stellaria nemorum l. from dolina mała łąka valley (tnp, pl) by wróblewski (1922b), and later cited by starmachowa (1963). however, kochman & majewski (1970) did not find it in the herbarium specimen. peronospora alta fuckel – plantago major l. tnp: pl – tatry zachodnie mts: dolina chochołowska valley; vii (kućmierz 1968). dolina ku dziurze valley at alt. 895 m; dolina białego valley at alt. 920 m; dolina kościeliska valley (stare kościeliska) at alt. 940 m; dolina olczyska valley at alt. 980 m; kuźnice village at alt. 1051 m; boczań mt. at alt. 1150 m, roadsides for all localities; viii (sałata et al. 1984). cited by sałata & mułenko (1996) and mułenko et al. (2004). peronospora boni-henrici gäum. [peronospora effusa auct. p.p.] – chenopodium bonus-henricus l. tnp: pl – tatry zachodnie mts: kuźnice village; vi (wróblewski 1922b). and at alt. 1053 m, roadside; viii (sałata et al. 1984). dolina chochołowska valley; vii (kućmierz 1968).cited by starmachowa (1963), kućmierz (1968), kochman & majewski (1970) and mułenko et al. (2004). peronospora conferta (unger) gäum. [peronospora alsinearum auct. p.p.] – cerastium fontanum baumg. tnp: pl – tatry zachodnie mts: kopa królowa mała mt. at alt. 1455 m, herbaceous plants in pinetum mugo carpaticum; giewont mt. at alt. 1850 m, calcareous grassland; viii (sałata et al. 1984). cited by mułenko et al. (2004). – cerastium holosteoides fr. emend. hyl. [cerastium fontanum baumg. subsp. triviale (link) jalas, cerastium vulgatum l.]. tnp: pl – tatry zachodnie mts: dolina jaworzynka valley at alt. 1080 m, grassland; przełęcz między kopami pass at alt. 1510 m, roadside; vi (mułenko et al. 1995). – cerastium tatrae borbás (cerastium raciborskii zapał.). tnp: pl – tatry zachodnie mts: mała dolinka pod giewontem valley; vii (wróblewski 1922b). cited by starmachowa (1963), kochman & majewski (1970), sałata & mułenko (1996) and mułenko et al. (2004). peronospora debaryi e. s. salmon & ware [peronospora urticae (lib.) de bary] – urtica urens l. tnp: pl – tatry zachodnie mts: kuźnice village (starmachowa 1963). cited by mułenko et al. (2004). peronospora erysimi gaüm. – erysimum hieraciifolium l. tnp: sk – vysoké tatry mts: between popradské pleso lake and popradské pleso railway station at alt. 1400 m; viii (müller 1980). cited by bacigálová (1999, on erysimum marschallianum andrz. ex m. bieb.). notes: according to marhold and hindák (1998) erysimum hieraciifolium l. is treated as a synonym of erysimum marschallianum andrz. ex bieb., however in the polish literature (mirek et al. 2002) the two species are treated separately. we cite the original record in this case. peronospora galii fuckel – galium anisophyllon vill. tnp: pl – tatry zachodnie mts: mały giewont mt. at alt. 1650 m, rock crevasse; viii (mułenko et al. 1995). cited by sałata & mułenko (1996) and mułenko et al. (2004). – galium schultesii vest. tnp: pl – tatry zachodnie mts: wąwóz kraków ravine at alt. 1018 m, spruce forest with abies alba; grzybowiec mt. at alt. 1175 m, spruce 14 w. mułenko et al. © the author(s) 2014 published by polish botanical society forest with abies alba; viii (sałata et al. 1984). dolina białego valley at alt. 970 m, herbaceous plants; vi (mułenko et al. 1995). cited by mułenko et al. (2004). peronospora grisea (unger) de bary – veronica beccabunga l. tnp: pl – tatry zachodnie mts: przełęcz iwaniacka pass; vii (kućmierz 1968). cited by mułenko et al. (2004). – veronica serpyllifolia l. tnp: pl – tatry wysokie mts: dolina roztoki valley; vii (wróblewski 1922b). cited by starmachowa (1963, as peronospora verna gäum.) and mułenko et al. (2004). ? peronospora hiemalis gäum. notes: the species was noted on ranunculus lanuginosus l. from dolina kościeliska valley (pl) by wróblewski (1922b), and later cited by starmachowa (1963) and mułenko et al. (2004). however, kochman & majewski (1970) did not find it in the herbarium specimen. peronospora knautiae fuckel – scabiosa lucida vill. tnp: pl – tatry zachodnie mts: giewont mt. – suchy żleb gully under wrótka pass at alt. 1380 m, calcareous grassland; viii (sałata et al. 1984). hala mała łąka mountain pasture (niżnia świstówka – below przełęcz kondracka pass) at alt. 1500 m and 1600 m, alpine grassland; ix (mułenko et al. 1995). cited by sałata & mułenko (1996) and mułenko et al. (2004). ? peronospora lepidii (mcalp.) g. w. wils. notes: the species was reported by wróblewski (1922b, as peronospora parasitica) on lepidium alpinum l. [hutchinsia alpina (l.) r. br.] from mała dolinka pod giewontem valley (pl). however, kochman & majewski (1970) did not find it in the herbarium specimen. peronospora mayorii gäum. – vicia cracca l. tnp: sk – vysoké tatry mts: popradské pleso railway station at alt. 1280 m; viii (müller 1980). cited by bacigálová (1999). peronospora myosotidis de bary – myosotis palustris (l.) l. emend. rchb. (myosotis scorpioides l.). tnp: pl – tatry wysokie mts: hala gąsienicowa mountain pasture at alt. 1530 m, herbaceous plants; vi (mułenko et al. 1995). cited by mułenko et al. (2004). peronospora phyteumatis fuckel – phyteuma spicatum l. tnp: pl – tatry zachodnie mts: dolina białego valley at alt. 910 m, spruce forest with abies alba; dolina spadowiec valley at alt. 915 m, spruce forest with abies alba; viii (sałata et al. 1984). cited by mułenko et al. (2004). peronospora potentillae de bary – potentilla aurea l. tnp: pl – tatry zachodnie mts: dolina chochołowska valley; vii (kućmierz 1968). dolina olczyska valley at alt. 950 m; stara polana glade at alt. 1060 m; dolina bystrej valley at alt. 1124 m; czoła jaworzyńskie hill at alt. 1195 m; czerwona przełęcz pass at alt. 1303 m; giewont mt. (piekło) at alt. 1405 m; kopa magury mt. at alt. 1560 m, grasslands for all localities; viii (sałata et al. 1984). cited by sałata & mułenko (1996) and mułenko et al. (2004). sk – vysoké tatry mts: mlynická dolina valley (near štrbské pleso lake) at alt. 1350 m, spruce forest; viii (müller 1980). bielovodská dolina valley; vii (skalický 1983). cited by bacigálová (1999). peronospora ranunculi gäum. [peronospora ficariae auct. p.p.] – ranunculus flammula l. notes: reported by kochman & majewski (1970) from tatra mts after wróblewski (1922b), however in the last paper it was located in zakopane town. microfungi of the tatra mts. 6. 15 © the author(s) 2014 published by polish botanical society – ranunculus lanuginosus → see notes on peronospora hiemalis. – ranunculus oreophilus m. bieb. tnp: pl – tatry zachodnie mts: nosal mt. at alt. 1150 m, alpine grassland; vi (mułenko et al. 1995). cited by mułenko et al. (2004). – ranunculus platanifolius → see peronospora alpicola. – ranunculus repens l. tnp: pl – tatry zachodnie mts: dolina kościeliska valley; viii (wróblewski 1922b). kuźnice village; giewont mt. – suchy żleb gully under wrótka pass (starmachowa 1963). cited by starmachowa (1963) and mułenko et al. (2004). peronospora rumicis corda – rumex acetosa l. tnp: pl – tatry zachodnie mts: dolina mała łąka valley; vi (wróblewski 1922b). dolina chochołowska valley; vii (kućmierz 1968). cited by starmachowa (1963) and mułenko et al. (2004). – rumex acetosella l. tnp: pl – tatry zachodnie mts: polana chochołowska glade; vii (kućmierz 1968). cited by mułenko et al. (2004, incorrectly reported after starmachowa 1963 instead of kućmierz l.c.). – rumex alpestris jacq. (rumex arifolius all.). tnp: pl – tatry zachodnie mts: mała dolinka pod giewontem valley; viii (wróblewski 1922b). cited by starmachowa (1963), kochman & majewski (1970), sałata & mułenko (1996) and mułenko et al. (2004). peronospora septentrionalis gäum. – cerastium cerastoides (l.) britton (cerastium trigynum vill.). tnp: sk – belianské tatry mts: kopské sedlo pass (picbauer 1933). cited by starmachowa (1963) and bacigálová [1999, on dichodon cerastoides (l.) rchb.]. peronospora sordida berk. – scrophularia scopolii hoppe. tnp: pl – tatry zachodnie mts: dolina strążyska valley; mała dolinka pod giewontem valley; dolina kościeliska valley; vi-vii (wróblewski 1922b). cited by starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). peronospora trifoliorum de bary [peronospora fulva syd.] – lathyrus pratensis l. tnp: pl – tatry zachodnie mts: dolina kościeliska valley (stare kościeliska) at alt. 960 m, meadow; vi (mułenko et al. 1995). cited by mułenko et al. (2004). – trifolium repens l. tnp: pl – tatry zachodnie mts: kuźnice village at alt. 1051 m, roadside; viii (sałata et al. 1984). cited by mułenko et al. (2004). peronospora violae de bary – viola biflora l. tnp: pl – tatry zachodnie mts: dolina strążyska valley; vi (wróblewski 1922b). cited by starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). plasmopara densa (rabenh.) j. schröt. – rhinanthus serotinus (schönh.) oborný [alectorolophus glaber (lam.) beck]. tnp: pl – tatry zachodnie mts: polana kalatówki mountain pasture (starmachowa 1963). cited by mułenko et al. (2004). plasmopara geranii-silvatici săvul. & o. săvul. – geranium palustre l. tnp: sk – západné tatry mts: jalovská dolina valley at alt. 740 m, roadside in spruce forest; vii (mułenko et al. 2008). 16 w. mułenko et al. © the author(s) 2014 published by polish botanical society – geranium pratense l. tnp: pl – tatry zachodnie mts: polana olczyska glade at alt. 1050 m, meadow; vi (mułenko et al. 1995, as plasmopara pusilla). cited by mułenko et al. (2008). sk – belianské tatry mts: dolina siedmich prameňov valley at alt. 900 m, roadside in spruce forest; vii (mułenko et al. 2008). – geranium sylvaticum l. tnp: pl – tatry zachodnie mts: polana kalatówki mountain pasture at alt. 1225 m, herbaceous plants; viii (sałata et al. 1984, as plasmopara pusilla). dolina olczyska valley (polana olczyska glade) at alt. 1059 m and 1115 m, meadow; dolina jaworzynka valley at alt. 1100 m, meadow; dolina miętusia valley at alt. 1120 m and 1180 m; ścieżka nad reglami tourist route (between dolina mała łąka valley and przysłop miętusi pass) at alt. 1158 m, meadow; przysłop miętusi pass at alt. 1165 m, meadow; dolina kościeliska valley (between ornak shelter house and iwaniacka przełęcz pass) at alt. 1220 m, herbaceous plants; tourist route between przysłop miętusi pass and małołączniak mt. at alt. 1342 m, 1362 m and 1394 m, spruce forest; giewont mt. – suchy żleb gully under wrótka pass at alt. 1350 m, herbaceous plants on screen slope; dolina tomanowa valley at alt. 1351 m, spruce forest; hala miętusia mountain pasture at alt. 1410 m and 1470 m; dolina mała łąka valley (between hala mała łąka mountain pasture and przełęcz kondracka pass) at alt. 1450 m, herbaceous plants; iwaniacka przełęcz pass at alt. 1460 m, herbaceous plants; vi-viii (mułenko et al. 2008). cited by mułenko et al. (2008). sk – západné tatry mts: javorová dolina valley (near tichá dolina valley) at alt. 1110 m, meadow; viii (voglmayr et al. 2006; mułenko et al. 2008). javorový žľab gully (from the side of tomanovská dolina valley) at alt. 1120 m; roháčska dolina valley (blue tourist route to spálena dolina valley) at alt. 1200 m; kamenistá dolina valley at alt. 1200 m and 1340 m; tichá dolina valley at alt. 1260 m; tomanovská dolina valley at alt. 1260 m; žiarska dolina valley (near žiarska chata shelter house) at alt. 1289 m and 1320 m, herbaceous plants; between žiarska chata shelter house and malé závraty cirque at alt. 1540 m, pinetum mughi carpaticum; red tourist route between pálenica and sivý vrch mt. at alt. 1640 m; viii (mułenko et al. 2008). vysoké tatry mts: tatranská lomnica town at alt. 860 m; dolina kežmarskej bielej vody valley at alt. 920 m, roadside in spruce forest and 1090 m, spruce forest; dolina bielych plies valley at alt. 1526 m, 1600 m and 1671 m, pinetum mughi carpaticum; vii-ix (mułenko et al. 2008). belianské tatry mts: dolina siedmich prameňov valley (below chata plesnivec shelter house) at alt. 1140 m and 1150 m, herbaceous plants; near chata plesnivec shelter house at alt. 1270 m; zadné meďodoly valley at alt. 1400 m and 1410 m, herbaceous plants; vii-ix (mułenko et al. 2008). – geranium sp. tnp: pl – tatry zachodnie mts: ścieżka nad reglami tourist route (between dolina mała łąka valley and dolina strążyska valley) at alt. 1240 m; ix (mułenko et al. 2008). sk – belianské tatry mts: monkova dolina valley at alt. 1650 m, mountain pasture, herbaceous plants; viii (mułenko et al. 2008). plasmopara nivea (unger) j. schröt. [plasmopara aegopodii (casp.) trotter, plasmopara umbelliferarum (casp.) schröt. ex wartenw.] – aegopodium podagraria l. tnp: pl – tatry zachodnie mts: dolina olczyska valley (starmachowa 1963). dolina chochołowska valley at alt. 1100 m, meadow; vii (mułenko et al. 1995). cited by mułenko et al. (2004). microfungi of the tatra mts. 6. 17 © the author(s) 2014 published by polish botanical society – anthriscus sylvestris (l.) hoffm. tnp: pl – tatry zachodnie mts: polana olczy-olczyska glade at alt. 1050 m, meadow; vi (mułenko et al. 1995). cited by mułenko et al. (2004). – mutellina purpurea (poir.) thell. [ligusticum mutellina (l.) crantz., meum mutellina (l.) gaertn.]. tnp: pl – tatry wysokie mts: morskie oko lake; viii (stec-rouppertowa 1936). cited by starmachowa (1963), kochman & majewski (1970) and mułenko et al. (2004). sk – vysoké tatry mts: temnosmrečinská dolina valley; viii (wróblewski 1922b). cited by starmachowa (1963) and bacigálová (1999, incorrectly from západné tatry mts). note: information on the species was also cited in a paper by sałata and mułenko (1996). plasmopara praetermissa voglmayr, faethi & constant. – geranium sylvaticum l. tnp: pl – tatry wysokie mts: tourist route between morskie oko lake and szpiglasowy wierch mt. at alt. 1600 m, pinetum mughi carpaticum; ix (mułenko et al. 2008). sk – západné tatry mts: zadná tichá dolina valley at alt. 1520 m, pinetum mughi carpaticum; vii (voglmayr et al. 2006; mułenko et al. 2008). tichá dolina valley (tourist route to kasprov vrch mt.) at alt. 1265 m, meadow; viii (mułenko et al. 2008). vysoké tatry mts: dolina bielych plies valley at alt. 1600 m, pinetum mughi carpaticum; zlomisková dolina valley at alt. 1640 m, herbaceous plants in pinetum mughi carpaticum; viii (mułenko et al. 2008). belianské tatry mts: dolina siedmich prameňov valley at alt. 1100 m; zadné meďodoly valley at alt. 1350 m, meadow; vii-viii (mułenko et al. 2008). – geranium sp. tnp: pl – tatry zachodnie mts: wielka polana małołącka glade at alt. 1161 m, meadow; vi (mułenko et al. 2008). plasmopara pusilla (de bary) j. schröt. – geranium pratense l. tnp: sk – vysoké tatry mts: tatranská lomnica town – centre at alt. 860 m and park near hotel morava at alt. 880 m and 900 m, roadside; stará lesná village – road between academia hotel and tatranská lomnica town at alt. 800 m, roadside; dolina kežmarskej bielej vody valley at alt. 1100 m; vii-viii (mułenko et al. 2008). – geranium sylvaticum l. tnp: pl – tatry zachodnie mts: giewont mt. – suchy żleb gully under wrótka pass at alt. 1385 m, herbaceous plants; viii (sałata et al. 1984). cited by mułenko et al. (2004). sk – vysoké tatry mts: between popradské pleso lake and popradské pleso railway station at alt. 1380 m, spruce forest; viii (müller 1980). cited by bacigálová (1999). notes: the herbarium specimen collected by müller was not reidentified. the plant may have been infected by plasmopara geranii-sylvatici. the data by sałata et al. (1984: suchy żleb gully, see above) are doubtful. the specimen was not preserved. it is likely to be plasmopara geranii-sylvatici sãvul. & o. sãvul. (mułenko et al. 2008). this species was also noted on geranium pratense l. (mułenko et al. 1995) and on geranium sylvaticum l. (sałata et al. 1984), and later cited by mułenko et al. (2004). the specimens were reidentified and included in plasmopara geranii-sylvatici. 18 w. mułenko et al. © the author(s) 2014 published by polish botanical society pythiales myzocytium megastomum de wild. – closterium lunula (müller) nitzsch. (algae). tnp: pl – tatry zachodnie mts: dolina gąsienicowa valley – toporowe stawy lakes; ix (kadłubowska 1999). acknowledgements. the study is part of two grants of the polish ministry of science and higher education (pl): no 2/04c/089/27 and no n/n304/172436, and also by vega grant agency (sk): no. 2/4032/04. references adamska i. 2001. microscopic fungus-like organisms and fungi of the słowiński national park (nw poland). ii. acta mycol. 36 (1): 31-65. bacigálová k. 1999. microscopic phytopathogenic fungi in high tatra mts. i. štúdie o tatranskom národnom parku 4 (37): 4170 (in slovak with english summary). bacigálová k., mułenko w., wołczańska a. 2005. parasitic microfungi of the tatra mts. 1. taphrinales. pol. bot. j. 50 (2): 185-207. choi y.j., thines m., piątek m., shin h.d. 2012. morphological evidence supports the existence of multiple species in pustula (albuginaceae, oomycota). nova hedwigia 94: 181-192. http://dx.doi. org/10.1127/0029-5035/2012/0094-0181 constantinescu m. 1991. an annotated list of peronospora names. thunbergia 15:1-110. danilkiewicz m. 1987. parasitic fungi of river bug valley. acta mycol. 23 (2): 37-80 (in polish with english abstract and summary). hazslinszky f. 1864. beiträge zur kenntnis der karpathenflora ix. brandpilze. verh. zool.-bot. ges. wien 14: 169-190. holmgren p. k., holmgren n. h., barnett l. c. 1990. index herbariorum. part i: the herbaria of the world. ed. 8. regnum veg. 120: 1-693. index fungorum, retrieved 2013-04-17 from http://www.indexfungorum.org/. kadłubowska j. 1999. rare species of fungi parasiting on algae. ii. parasites of desmidiaceae. acta mycol. 34 (1): 51-54. kirk p.m., cannon p.f., minter d.w., stalpers j.a. 2008. ainsworth, bisby’s dictionary of the fungi. 10th edition. cabi international. kochman j., majewski t. 1970. glonowce (phycomycetes), wroślikowe (peronosporales). 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(eds). 1998. zoznam nižších a vyšších rastlín slovenska, pp. 687. veda, bratislava, 687 pp. microfungi of the tatra mts. 6. 19 © the author(s) 2014 published by polish botanical society mirek z., głowaciński z., klimek k., piękoś-mirkowa h. (eds) 1996. nature of the tatra national park. tatry i podtatrze 3: 787 pp. wyd. tatrzański park narodowy, kraków-zakopane (in polish with english summaries). mirek z., piękoś-mirkowa h., zając a., zając m. 2002. flowering plants and pteridophytes of poland. a checklist. w. szafer institute of botany, polish academy of sciences, kraków. moesz g. 1930. pilze aus dem norden ungarns. folia cryptog. 1 (7): 795-816. müller j. 1980. contribution towards the knowlwdge of rusts, smuts and downy mildew in high tatras. biologia (bratislava) 35 (7): 497-504 (in czech with russian summary and english abstract). mułenko w. 1988a. the microscopic pathogenic fungi of the łęczna-włodawa lake district. i. the occurrence of pathogenic fungi in plant communities and theirs phenology. acta mycol. 24 (1): 3-49 (in polish with english summary). mułenko w. 1988b. the microscopic pathogenic fungi of the łęczna-włodawa lake district. ii. acta mycol. 24 (2): 125-171 (in polish with english summary). mułenko w., bacigálová k. 2005. parasitic microfungi of the tatra mts. 2. pseudocercosporella tatrensis sp. nov. on aconitum firmum subsp. firmum from poland. pol. bot. j. 50 (2): 209-212. mułenko w., bacigálová k., kozłowska m. 2005. parasitic microfungi of the tatra mts. 4. melampsoridium hiratsukanum (urediniomycetes). pol. bot. stud. 22: 399-405. mułenko w., bacigálová k., wołczańska a., świderska u., mamczarz m. 2008. parasitic microfungi of the tatra mts. 5. plasmopara representatives on the species of geranium. biologia (bratislava) 63(3): 302-306. mułenko w., kozłowska m., sałata b. 2004. microfungi of the tatra national park. a checklist. biodiversity of the tatra national park. vol. 1: pp. 68. w. szafer institute of botany, polish academy of sciences, kraków. mułenko w., sałata b., wołczańska a. 1995. mycological notes from the tatra national park. ii. acta mycol. 30 (1): 65-79. mułenko w., wołczańska a., bacigálová k. 2006. taphrinales (ascomycota) in the tatra mountains. analysis of occurrence. (in:) z. mirek, b. godzik (eds). przyroda tatrzańskiego parku narodowego a człowiek. 2005. tatrzański park narodowy na tle innych górskich terenów chronionych. vol. ii. nauki biologiczne. kraków-zakopane 2006: 29-33. wyd. tatrzańskiego parku narodowego (in polish with english summary). namysłowski b. 1914. śluzowce i grzyby galicyi i bukowiny. pam. fizjogr. 22: 1-151. radwańska-paryska z., paryski w. h. 2004. wielka encyklopedia tatrzańska. wydawnictwo górskie, poronin, 1555 pp. picbauer r. 1933. addenda ad floram čechoslovakiae mycologicam. vii. práce morav. přír. společn. 8 (8): 1-20. plaats-nitterink, a.j.van der.1981. monograph of the genus pythium. stud. mycol., 21: 1-244. centraalbureau voor schimmelcultures, utrecht, nederlands. romaszewska-sałata j., sałata b., mułenko w. 1986. on some interesting representatives of peronosporales and erysiphales collected recently in poland. folia soc. sci. lubl., biologia 28 (1): 11-18 (in polish with english and russian summaries). rouppert k. 1912. pilze aus der tatra und den westlichen beskiden. spraw. kom. fizjogr. 46: 80-100 (in polish with german summary). ruszkiewicz-michalska m. 2006. phytoparasitic micromycetes in plant communities of the wyżyna częstochowska upland. monogr. bot. 96: 1-140 (in polish with english summary). sałata b., mułenko w. 1996. microscopic phytopathogenic fungi. (in:) z. mirek, z. głowaciński, k. klimek, h. piękoś-mirkowa (eds). nature of the tatra national park. tatry i podtatrze 3: 393-404. tatrzański park narodowy, zakopane-kraków (in polish with english summary). sałata b., romaszewska-sałata j., mułenko w. 1984. mycological notes from the polish tatra national park. acta mycol. 20(1): 13-21 (in polish with english abstract). saharan g. s., verna p. r. noshaat n. j. 1997. monograph on downy mildew of crucifers. technical bulletin – agriculture and agri-food, canada, 208 pp. skalický v. 1983. the revision of species of the genus peronospora on host plant of the family rosaceae with respect to central european species. folia geobot. phytotax. 18: 71-101. starmachowa b. 1963. les champignons parasitaires des tatras. monogr. bot. 15: 153-294 (in polish with french summary). stec-rouppertowa w. 1936. zapiski mikologiczne. spraw. kom. fizjogr. 70: 149-172. 20 w. mułenko et al. © the author(s) 2014 published by polish botanical society szulczewski j. w. 1930. die gallen des polnischen tatragebriges. spraw. kom. fizjogr. 64: 1-11 (in polish with german summary). thines m., spring o. 2005. a revision of albugo (chromista, peronosporomycetes). mycotaxon 92: 443458. urban z. 1952. hrdze a sneti v temnosmrečinovej doline vo vysokých tatrách. slovenská akadémie vied a umeni, bratislava (in czech with russian summary). voglmayr h., fatehi j., constantinescu o. 2006. revision of plasmopara (chromista, peronosporales) parasitic on geraniaceae. mycol. res. 110: 633-645. http://dx.doi.org/10.1016/j.mycres.2006.03.005 vološčuk i. (ed.). 1994. tatranský národný park. biosférická rezervácia. vyd. gradus, martin. wołczańska a., mułenko w., oklejewicz k., bacigálová k. 2008. parasitic microfungi of the tatra mts. 3. spermosporina gymnadeniae – new species for slovakia and new records in poland. biologia 63(1): 50-52. wróblewski a. 1918. beitrag zur kenntnis der pilzflora westgaliziens. spraw. kom. fizjogr. 52: 122-127 (in polish with german summary). wróblewski a. 1920. grzyby zbioru józefa krupy. spraw. kom. fizjogr. 53-54: 83-94. wróblewski a. 1922a. les champignons recueillés par j. krupa. kosmos 47 (1-3): 39-51 (in polish with french summary). wróblewski a. 1922b. wykaz grzybów zebranych w latach 1913-1918 z tatr, pienin, beskidów wschodnich, podkarpacia, podola, roztocza i innych miejscowości. i. phycomycetes, ustilaginaceae, uredinales i basidiomycetes. spraw. kom. fizjogr. 55-56: 1-50. wróblewski a. 1925. champignons recueillis par m. raciborski dans les environs de cracovie et dans le tatra en 1883 et 1890. acta soc. bot. pol. 3 (1): 29-41 (in polish with french summary). grzyby mikroskopijne tatr. 6. organizmy grzybopodobne: albuginales, peronosporales i pythiales streszczenie masyw tatr, jeden z najciekawszych pod względem przyrodniczym regionów europy środkowej, został dość dobrze poznany pod wieloma względami, np. geologicznym, hydrologicznym, geobotanicznym, florystycznym. w dalszym ciągu jednak nie jest wystarczająco poznany pod względem mykologicznym, zwłaszcza w zakresie grzybów mikroskopijnych oraz organizmów grzybopodobnych. trwające od kilku lat prace mające na celu podsumowanie dotychczasowego stanu zbadania tej grupy przyczyniły się do opublikowania kilku prac syntetyczno-analitycznych poświęconych kolejnym jednostkom taksonomicznym – rzędom, rodzajom czy gatunkom. w obecnej pracy przygotowano wykaz organizmów grzybopodobnych należących do trzech rzędów w obrębie typu oomycota – albuginales, peronosporales oraz pythiales, podawanych dotychczas w granicach parków narodowych obu części tatr – polskiej i słowackiej. zamieszczono w nim gatunki znane przede wszystkim z literatury, uzupełniając część informacji o dane szczegółowe pochodzące z kolekcji zielnikowych. pierwsze dane o organizmach grzybopodobnych tatr pojawiły się pod koniec xix wieku w pracach krupy (1886, 1888). do chwili obecnej, a więc przez okres ponad 120 lat, publikacje dotyczące oomycota były jednak nieliczne, a informacje na temat ich występowania bardzo ogólne. na całym obszarze tatr stwierdzono dotychczas zaledwie 32 gatunki na 57 gatunkach roślin, w tym dwa gatunki z rzędu albuginales (rodzaje: albugo i pustula, na 9 gatunkach roślin), 29 gatunków z rzędu peronosporales (rodzaje: bremia, hyaloperonospora, peronospora oraz plasmopara, na 49 gatunkach roślin), a także jeden gatunek z rzędu pythiales (rodzaj: myzocytium, na jednym gatunku glonu). po polskiej stronie tatr zebrano 29 gatunków, przy microfungi of the tatra mts. 6. 21 © the author(s) 2014 published by polish botanical society czym aż 26 w tatrach zachodnich. po słowackiej stronie tatr zanotowano zaledwie 10 gatunków, z czego najwięcej – 9 w tatrach wysokich (vysoké tatry). wszystkie zebrano na 185 stanowiskach, przy czym 135 przypada na tatry polskie (73 %), w tym aż 126 na polskie tatry zachodnie. z kolei po stronie słowackiej tatr najwięcej danych pochodzi z tatr wysokich (21 stanowisk). stan znajomości oomycota w tatrach jest więc bardzo słaby, a informacje o ich rozmieszczeniu rozproszone, wręcz przypadkowe. w przyszłości niezbędne są badania znacznie bardziej intensywne, przede wszystkim systematyczne, a obecny syntetyczny wykaz może być punktem wyjścia do ich rozpoczęcia. 2014-06-30t21:04:51+0200 polish botanical society 2014-11-20t23:11:46+0000 polish botanical society 2014-11-20t23:15:45+0000 polish botanical society 2014-11-20t23:16:18+0000 polish botanical society 2014-11-20t23:20:49+0000 polish botanical society 2014-11-20t23:14:35+0000 polish botanical society 2014-11-20t23:17:00+0000 polish botanical society 2014-11-20t23:18:40+0000 polish botanical society 2014-11-20t23:19:29+0000 polish botanical society new records of parmelia ernstiae and p. serrana (ascomycota, parmeliaceae) in poland 1 of 6published by polish botanical society acta mycologica short communication new records of parmelia ernstiae and p. serrana (ascomycota, parmeliaceae) in poland emilia ossowska1*, katarzyna szczepańska2, maria kossowska3 1 department of plant taxonomy and nature conservation, university of gdańsk, wita stwosza 59, 80-308 gdańsk, poland 2 department of botany and plant ecology, wrocław university of environmental and life sciences, pl. grunwaldzki 24a, 50-363 wrocław, poland 3 laboratory of lichenology, department of botany, institute of environmental biology, university of wrocław, kanonia 6/8, 50-328 wrocław, poland * corresponding author. email: emilia.ossowska@biol.ug.edu.pl abstract the paper reports new localities of lichens parmelia ernstiae and p. serrana in poland. both species are reported for the first time from the polish part of the sudety mountains (southwestern part of the country). the new localities were found in the area of the sowie mountains (central part of the sudety mts) as well as śnieżnik massif and bialskie mountains (eastern part of the sudety mts). most of the recorded specimens were associated with the bark of acer platanoides and a. pseudoplatanus. parmelia ernstiae and p. serrana are still poorly known species in poland and often confused with similar taxon p. saxatilis. therefore the morphological characteristics, chemical properties, ecology and distribution of these species are presented and briefly discussed. keywords lichenized fungi; parmelioid lichens; biodiversity; distribution; sudety mts introduction the genus parmelia belongs to parmeliaceae, the largest family of lichenized fungi [1,2]. generally, members of this genus are characterized by foliose, loosely to closely adnate thalli. upper surface is grey, whitish grey to greyish brown, smooth to foveolate, sometimes pruinose, with white, linear pseudocyphellae. lower surface is black with simple or branched rhizines. soredia and/or isidia are developed on the upper surface in most species [3–5]. in terms of chemistry, atranorin constitute the main cortical secondary metabolite. the species produce also salazinic, consalazinic, lobaric, protocetraric and galbanic acids as well as fatty acids (e.g., lichesterinic acid) in various combinations [3–5]. seven species of parmelia have been reported from poland so far, i.e., parmelia discordans nyl., p. ernstiae feurer & a. thell, p. omphalodes (l.) ach., p. saxatilis (l.) ach., p. serrana a. crespo, m.c. molina & d. hawksw., p. submontana nádv. ex hale, and p. sulcata taylor. [6–11]. they differ primarily in terms of chemical properties (e.g., production of fatty acids) and also some morphological features, such as pruinosity of upper surface and the shape of rhizines. parmelia ernstiae and p. serrana have been recognized in poland relatively recently, in 2012 and 2014, respectively [9,11]. therefore, their detail distribution and frequency of occurrence in poland is still poorly known. historical and contemporary reports on lichens from the sudety mountains are quite numerous (e.g., [12–24]), but the presence of these two parmelia species in the polish part of this mountain range was not documented until now. doi: 10.5586/am.1065 publication history received: 2015-07-21 accepted: 2015-11-30 published: 2016-01-29 handling editor tomasz leski, institute of dendrology of the polish academy of sciences, poland authors’ contributions eo, ks: field and herbarium research; eo, ks: determination of specimens; eo, ks: chromatographic analyses; eo, ks, mk: writing the manuscript; eo, mk: prepartation of the distribution map funding the research was supported by the polish ministry of science and higher education, project no. 2012/07/n/nz8/00061 granted to eo. competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation ossowska e, szczepańska k, kossowska m. new records of parmelia ernstiae and p. serrana (ascomycota, parmeliaceae) in poland. acta mycol. 2015;50(2):1065. http://dx.doi. org/10.5586/am.1065 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:emilia.ossowska%40biol.ug.edu.pl?subject=new%20records%20of%20parmelia%20ernstiae%20and%20p.%20serrana%20%28ascomycota%2c%20parmeliaceae%29%20in%20poland http://dx.doi.org/10.5586/am.1065 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1065 http://dx.doi.org/10.5586/am.1065 2 of 6© the author(s) 2016 published by polish botanical society acta mycol 50(2):1065 ossowska et al. / new records of parmelia ernstiae and p. serrana this paper bridge the gap in their distribution in poland and presents new localities, which have been recently discovered in the sudety mts. additionally notes about morphology, chemistry, distribution and habitat requirements of these species are also provided. material and methods the present study is based on material which is available in polish lichen herbaria: lod, ugda, wrsl and herb. szczepańska. the morphology of the specimens were examined under a stereomicroscope for: color and structure (pruinoisity or epruinoisity) on the upper surface, shape of lobes, shape of rhizines, location of pseudocyphellae, shape and location of isidia. brief descriptions of the species provided in this paper are based on personal observations. the lichen substances were examined by thin-layer chromatography (tlc) in solvent a and c; the methodology follows orange et al. [25] and kubiak and kukwa [26]. localities of examined specimens are mapped according to the atpol grid square system [27], modified by cieśliński and fałtynowicz [28]. results and discussion parmelia ernstiae feurer & a. thell mitt. inst. allg. bot. hamburg 30–32: 52. 2002. morphology. parmelia ernstiae is distinguished by the strong pruniose upper surface, isidia and short, broad and not overlapping lobes. pseudocyphellae are short, white, linear and narrow. in the central parts of the thalli cylindrical isidia are developed. additionally, small lobulae are often present on the upper surface. lower surface is black with simple rhizines (see also [4,5,29,30]). chemistry. the species is characterized by the production of atranorin, salazinic, consalazinic, protocetraric and lobaric acids as well as fatty acids (e.g., lichesterinic and protolichesterinic) [5,29,30]. in examined specimens all substances, except protocetraric, isonephrosterinic and nephrosterinic acids (probably due to their low quantity) were detected (see also [9,10]). notes. parmelia ernstiae is distinguished from morphologically similar taxa by the strong pruniose upper surface and isidia. parmelia saxatilis is softly pruinose whereas p. serrana has epruinose upper surface. parmelia ernstiae can be also easily separated from p. saxatilis and p. serrana based on the chemical properties of the species. it produces lobaric acid (absent in p. serrana) and the fatty acids (absent in p. saxatilis) [5,29,30]. habitat requirements. parmelia ernstiae is typically corticolous lichen. in the sudety mts it was found on the bark of deciduous trees: acer pseudoplatanus and a. platanoides. general distribution. generally, parmelia ernstiae is known from europe, asia and africa. it has been reported from: austria, belgium, bosnia and herzegovina, bulgaria, the czech republic, denmark, estonia, finland, france, germany, greece, great britain, ireland, lithuania, luxemburg, the netherlands, slovenia, sweden, spain [5,8,29–35]. outside europe it has been reported from nw part of russia, algeria and the canary islands [8,31]. distribution in poland. the species was reported firstly from the drawskie lakeland [9] and later from the elbląska high plain and the sławieńska plain [10]. eight 3 of 6© the author(s) 2016 published by polish botanical society acta mycol 50(2):1065 ossowska et al. / new records of parmelia ernstiae and p. serrana localities of p. ernstiae in the sudety mts are presented in this paper. the localities are situated in the sowie mountains as well as in the śnieżnik massif and bialskie mountains (fig. 1). specimens examined. eb-95: sudety mts, sowie mts, near kamionki residential, on acer sp., 24 mar. 2015, leg. e. ossowska, k. szczepańska, a. szczepański (ugda-l); sudety mts, sowie mts, on acer sp., 24 mar. 2015, leg. e. ossowska, k. szczepańska, a. szczepański (ugda-l). fb-38: sudety mts, bialskie mts, nowy gierałtów village, on acer platanoides, 27 jul. 2003, leg. k. szczepańska (wrsl-512). fb-47: sudety mts, śnieżnik massif, wilczka stream valley, on acer pseudoplatanus, 10 jul. 2003, leg. k. szczepańska (wrsl-2573). fb-48: sudety mts, bialskie mts, śnieżna białka nature reserve, on tree, 23 aug. 1993, leg. e. kozioł (wrsl); sudety mts, bialskie mts, puszcza jaworowa nature reserve, on acer pseudoplatanus, 8 jul. 2003, leg. k. szczepańska (wrsl-1030); sudety mts, bialskie mts, młynkówka stream valley, on acer platanoides, 6 aug. 2003, leg. k. szczepańska (wrsl-352); sudety mts, śnieżnik massif, road below leje, alt. ca. 760 m, on acer pseudoplatanus, 27 may 2004, leg. k. szczepańska 121 (herb. szczepańska). fb-56: sudety mts, śnieżnik massif, road between pisary and potoczek villages, on acer platanoides, 21 aug. 2003, leg. k. szczepańska (wrsl-1129). parmelia serrana a. crespo, m.c. molina & d. hawksw. lichenologist 36(1): 48 (2004). morphology. parmelia serrana is distinguished by epruinose upper surface (occasionally the lobes are softly pruinose) with short, broad and sometimes overlapping lobes. white, linear or irregular pseudocyphellae are developed on the upper surface and also on the margins of lobes. cylindrical isidia are concentrated along ridges of the thalli. lower surface is black with simple or branched rhizines (see also [4,5,33]). chemistry. the species is characterized by the production of atranorin, salazinic, consalazinic and protocetraric acids as well as fatty acids, e.g., lichesterinic and protolichesterinic. in examined material from sudety mts protocetraric, nephrosterinic and isonephrosterinic acids were not confirmed, probably due to their low quantity (see also [11,33]). notes. parmelia serrana can be distinguished from p. ernstiae by epruinose upper surface and isidia concentrated to the ridges. the species can be also confused with p. saxatilis which has softly pruinose upper surface and sublinear lobes. moreover, those three species are differ in terms of chemical properties. parmelia serrana produces fatty acids (absent in p. saxatilis) and does not produce lobaric acid (present in p. saxatilis and p. ernstiae) [5,29,33]. habitat requirements. parmelia serrana is corticolous lichen and all specimens found in the sudety mts were growing on the bark of acer pseudoplatanus and fraxinus excelsior. fig. 1 distribution of parmelia ernstiae in poland. 1 – localities reported by kukwa et al. [9] and ossowska, kukwa [10]; 2 – new localities. 4 of 6© the author(s) 2016 published by polish botanical society acta mycol 50(2):1065 ossowska et al. / new records of parmelia ernstiae and p. serrana general distribution. parmelia serrana has been reported from austria, germany, finland, spain, sweden and ukraine [5,9,33,35]. the species is known also from nw part of russia and the canary islands [33,35]. distribution in poland. the species was known only from the southeastern and northern part of poland until now [11] and six localities from southwestern poland presented here are in addition to its distribution in the country (fig. 2). specimens examined. ea-78: sudety mts, karkonosze mts, szklarska poręba town, kilińskiego street, on fraxinus excelsior, 3 may 1998, leg. m. ratajczak (lod10429). eb-95: sudety mts, sowie mts, near kamionki residential, on acer sp., 24 mar. 2015, leg. e. ossowska, k. szczepańska, a. szczepański (ugda-l); sudety mts, sowie mts, near kamionki residential, on acer sp., 24 mar. 2015, leg. e. ossowska, k. szczepańska, a. szczepański (ugdal); sudety mts, sowie mts, by the road to walim village, on branch ot tree, 24 mar. 2015, leg. e. ossowska, k. szczepańska, a. szczepański (ugda-l). fb-47: sudety mts, śnieżnik massif, road under leje, on acer pseudoplatanus, 27 may 2004, leg. k. szczepańska (wrsl-440). fb-48: sudety mts, bialskie mts, rude krzyże mt, mixed forest, on acer pseudoplatanus, 28 may 2004, leg. k. szczepańska (wrsl-1599). acknowledgments we are grateful to the all curators of polish herbaria for the loan of the specimens for the study and two anonymous reviewers for comments on the manuscript. references 1. crespo a, kauff f, divakar pk, del prado r, pérez-ortega s, amo de paz g, et al. phylogenetic generic classification of parmelioid lichens (parmeliaceae, ascomycota) based on molecular, morphological and chemical evidence. taxon. 2010;59(6):1735–1753. 2. thell a, crespo a, divakar pk, kärnefelt i, leavitt sd, lumbsch ht, et al. a review of the lichen family parmeliaceae – history, phylogeny and current taxonomy. nord j bot. 2012;30(6):641–664. http://dx.doi.org/10.1111/j.1756-1051.2012.00008.x 3. hale me. a monograph of the lichen genus parmelia acharius sensu stricto (ascomycotina: parmeliaceae). washington, dc: smithsonian institution press; 1987. (smithsonian contributions to botany; vol 66). 4. louwhoff shjj, purvis ow, james pw. parmelia ach. (1803). in: smith cw, aptroot a, cooppis bj, fletcher a, gilbert ol, james pw, et al., editors. the lichens of great britain and ireland. london: british lichen society; 2009. p. 651–654. 5. thell a, thor g, ahti t. parmelia ach. in: thell a, moberg r, editors. nordic lichen flora. 4. uddevalla: nordic lichen society; 2011. p. 83–90. 6. nowak j, tobolewski z. porosty polskie. warszawa: polska akademia nauk, instytut botaniki; 1975. 7. fałtynowicz w. the lichens, lichenicolous and allied fungi of poland – an annotated checklist. cracow: w. szafer institute of botany, polish academy of sciences; 2003. fig. 2 distribution of parmelia serrana in poland. 1 – localities reported by ossowska et al. [11]; 2 – new localities. http://dx.doi.org/10.1111/j.1756-1051.2012.00008.x 5 of 6© the author(s) 2016 published by polish botanical society acta mycol 50(2):1065 ossowska et al. / new records of parmelia ernstiae and p. serrana 8. hawksworth dl, blanco o, divakar pk, ahti t, crespo a. a first checklist of parmelioid and similar lichens in europe and some adjacent territories, adopting revised generic circumscriptions and with indications of species distributions. lichenologist. 2008;40(1):1– 21. http://dx.doi.org/10.1017/s0024282908007329 9. kukwa m, łubek a, szymczyk r, zalewska a. seven lichen species new to poland. mycotaxon. 2012;120(1):105–118. http://dx.doi.org/10.5248/120.105 10. ossowska e, kukwa m. nowe stanowiska porostu parmelia ernstiae feuerer & a. thell w polsce. acta bot cassub. 2012;11:195–199. 11. ossowska e, szymczyk r, bohdan a, kukwa m, the lichen family parmeliaceae in poland. iii. parmelia serrana, new to poland. acta soc bot pol. 2014;83(1):81–84. http://dx.doi. org/10.5586/asbp.2014.006 12. flotow j. lichenes florae silesiae i. jahresbericht der schlesischen ge-sellschaft für vaterländische kultur. 1850;27:98–135. 13. flotow j. lichenes florae silesiae ii. jahresbericht der schlesischen ge-sellschaft für vaterländische kultur. 1851;28:115–143. 14. körber g. systema lichenum germaniae. die flechten deutschlands (insbesondere schlesiens). breslau: trevendt & granier; 1855. 15. körber g. parerga lichenologica. ergänzungen zum systema lichenum germaniae. breslau: e. trevendt & granier; 1865. 16. stein b. flechten. breslau: j.u. kern; 1897. [cohn’s kryptogamen-flora von schlesien; vol 2(2)]. 17. stein b. nachträge zur flechtenflora schlesien. jahresbericht der schlesischen gesellschaft für vaterländische kultur. 1889;66:142–149. 18. eitner e. nachträge zur flechtenflora schlesiens. jahresbericht der schlesischen gesellschaft für vaterländische kultur. 1896;73:2–26. 19. eitner e. ii nachtrag zur schlesischen flechtenflora. jahresbericht der schlesischen gesellschaft für vaterländische kultur. 1901;78:5–27. 20. eitner e. dritten nachtrag zur schlesischen flechtenflora. jahresbericht der schlesischen ge-sellschaft für vaterländische kultur. 1911;88(1):20–60. 21. tobolewski z. porosty gór stołowych. poznań: państwowe wydawnictwo naukowe; 1955. (prace komisji biologicznej; vol 16). 22. fabiszewski j. porosty śnieżnika kłodzkiego i gór bialskich. warszawa: pwn; 1968. (monographiae botanicae; vol 26). 23. kossowska m. lichens growing on calcareous rocks in the polish part of the sudety mountains. wrocław: zakład bioróżnorodności i ochrony szaty roślinnej, instytut bilogii roślin uniwersytetu wrocławskiego; 2008. (acta botanica silesiaca, monographiae; vol 3). 24. szczepańska k. new lichens and lichenicolous fungi of the polish sudety mountains. pol bot j. 2007;52(2):165–170. 25. orange a, james pw, white fj. microchemical methods for the identification of lichens. london: british lichen society; 2001. 26. kubiak d, kukwa m. chromatografia cienkowarstwowa (tlc) w lichenologii. in: dynowska m, ejdys e, editors. mikologia laboratoryjna. przygotowanie materiału badawczego i diagnostyka. olsztyn: wydawnictwo uwm; 2011. p. 176–190. 27. zając a. atlas of distribution of vascular plants in poland (atpol). taxon. 1978;27(5– 6):481–484. http://dx.doi.org/10.2307/1219899 28. cieśliński s, fałtynowicz w. note from editors. in: cieśliński s, fałtynowicz w, editors. atlas of the geographical distribution of lichens in poland. cracow: w. szafer institute of botany, polish academy of sciences; 1993. p. 7–8. (vol 1). 29. feuerer t, thell a. parmelia ernstiae – a new macrolichen from germany. mitt inst allg bot hamb. 2002;30–32:49–60. 30. thell a, hansene es, kärnefelt i, feuerer t. the distribution of parmelia ernstiae in denmark. bibl lichenol. 2007;96:244–248. 31. serusiaux e, diedrich p, ertz d, van den boom p. new or interesting lichens and lichenicolous fungi from belgium, luxembourg and northern france. ix. lejeunia. 2003;173:1–48. 32. thell a. parmelia ernstiae – new to the nordic lichen flora. graph scr. 2003;14:10. 33. molina mc, crespo a, blanco o, lumbsch ht, hawksworth dl. phylogenetic http://dx.doi.org/10.1017/s0024282908007329 http://dx.doi.org/10.5248/120.105 http://dx.doi.org/10.5586/asbp.2014.006 http://dx.doi.org/10.5586/asbp.2014.006 http://dx.doi.org/10.2307/1219899 6 of 6© the author(s) 2016 published by polish botanical society acta mycol 50(2):1065 ossowska et al. / new records of parmelia ernstiae and p. serrana relationships and species concepts in parmelia s. str. (parmeliaceae) inferred from nuclear its rdna and β-tubulin sequences. lichenologist. 2004;36(1):37–54. http://dx.doi. org/10.1017/s0024282904013933 34. otte v. noteworthy lichen records for bulgaria. abh ber naturkundemus görlitz. 2005;77(1):77–86. 35. thell a, elix ja, feuerer t, hansene e, kärnefelt i, schuler n, et al. notes on the systematics, chemistry and distribution of european parmelia and punctelia species (lichenized ascomycetes). sauteria. 2008;15:545–559. http://dx.doi.org/10.1017/s0024282904013933 http://dx.doi.org/10.1017/s0024282904013933 abstract introduction material and methods results and discussion parmelia ernstiae feurer & a. thell parmelia serrana a. crespo, m.c. molina & d. hawksw. acknowledgments references 2016-01-25t16:55:02+0000 piotr otręba 2014-09-06t20:22:16+0200 polish botanical society non-lipophilic mycobiota of human skin 1 of 5published by polish botanical society acta mycologica original research paper non-lipophilic mycobiota of human skin katarzyna talaga, paweł krzyściak* department of mycology, chair of microbiology, jagiellonian university medical college, czysta 18, 31-121 cracow, poland * corresponding author. email: pawel.krzysciak@uj.edu.pl abstract the human skin is inhabited by many species of bacteria and fungi, which are its natural microbiota. fungi colonizing the skin, including those causing disease, characterized by great variety and variability, can be influenced by various factors. the purpose of this study was to investigate the composition of the non-lipiddependent fungal microbiota of skin, including the presence of species potentially pathogenic for humans. fifty-six volunteers of both sexes aged 22–78 were subjected to the study. swabs were taken from the face, chest, back and interdigital spaces of hands. mycobiota isolated proved to vary both in terms of the location of occurrence and gender of patients. interdigital spaces of hands, dominated by yeasts, constitute a location on human skin most contaminated with fungi. molds were more often isolated from the face and chest. the back was the least contaminated location. there was no difference in fungal incidence in relation to sex. keywords mycobiota; yeasts; microbial diversity; molds introduction the human skin is inhabited by many species of bacteria and fungi, which are its natural microbiota. these organisms perform many beneficial functions. among others, through competition, they do not allow pathogenic species to colonize the skin [1,2]. mycobiota of skin is dominated mainly by lipophilic fungi of the genus malassezia. additionally, other species of yeast, mainly of the genus candida and cryptococcus, may exist on the skin. species composition of skin mycobiota is characterized by great diversity and variability, which can be influenced by various factors such as age, occupation, and the climate of the place of residence [3–5]. monitoring of currently dominant fungal species belonging to the natural microbiota as well as the frequency and location of infection caused by these fungi is important for epidemiological studies. collecting these data allows drawing conclusions about ongoing changes and forecasting changes to mycobiota in the near future. the aim of this study was to determine settlement of human’s skin by non-lipophilic fungi. material and methods the study involved 56 generally healthy (no responding symptoms of fungal infection of skin and antifungal therapy) volunteers (41 women and 15 men) aged over 18 years. the mean age was 43.7 for women ±13.5 years (range 22–78 years, median = 42 years), and for men 48.9 ±15.7 years (range 17–75, median = 50). doi: 10.5586/am.1068 publication history received: 2015-09-22 accepted: 2016-01-08 published: 2016-01-29 handling editor tomasz leski, institute of dendrology of the polish academy of sciences, poland authors’ contributions pk contributed to the idea of research and collected the samples; both authors reviewed other studies and analyzed the data; both authors were involved in drafting and revising the manuscript; both authors approved the final version of the manuscript funding this study was founded from subsidies for the development of young scientist awarded by the polish ministry of science and higher education for jagiellonian university medical college (internal designation k/ dsc/001394). competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation talaga k, krzyściak p. nonlipophilic mycobiota of human skin. acta mycol. 2015;50(2):1068. http://dx.doi. org/10.5586/am.1068 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:pawel.krzysciak%40uj.edu.pl?subject=non-lipophilic%20mycobiota%20of%20human%20skin http://dx.doi.org/10.5586/am.1068 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1068 http://dx.doi.org/10.5586/am.1068 2 of 5© the author(s) 2016 published by polish botanical society acta mycol 50(2):1068 talaga and krzyściak / non-lipophilic mycobiota of skin the study was conducted in 2012–2014. swabs were taken from nasolabial folds, the area of the breastbone, the back and interdigital spaces of hands for every study participant. then the swabs were immediately cultured on sabouraud glucose agar (sga) with chloramphenicol and incubated at 27°c for 2–3 weeks and periodically checked. the growing colonies were subcultured on appropriate media (rice agar for yeasts and sporulation enhancement medium for molds, i.e., czapek–dox agar). the fungal identification to the genus level was based on identification keys according to atlas of clinical fungi (2nd ed.) [6]. for yeasts, in some cases, species identification was confirmed by api 20c aux strips (biomérieux) according to manufacturer’s procedure. for the mold colony, we performed adhesive tape slides in lactophenol cotton blue. in cases in which this method was not enough to id genera, we performed slide cultures. for molds colony we perform scotch type slides in lactophenol cotton blue in cases that this method was not enough to id genera we perform slide culture. the study was approved by the bioethical committee of jagiellonian university (no. kbet/164/b/2012). statistical analysis was performed using r language and environment for statistical computing software [7]. the significance level for all statistical tests was set at alpha (p) ≤ 0.05. results a positive culture from at least one location was obtained from 38 participants (n = 27; 65.8% of women and n = 11; 73.3% of men). there were no differences in the overall incidence of colonization between the sexes (fisher’s exact test, p = 0.7508). altogether, 72 (22.6%) positive cultures were obtained from 223 swabs (36 strains of yeast-like fungi and 30 of molds; 10 cultures showed mixed mycobiota – yeasts and molds). no fungi were isolated from any location in 18 participants. in 19 patients, a few species of fungi were isolated simultaneously from a single location. further results are given in tab. 1. the most frequently cultured yeasts in all locations were representatives of the genus candida. among the molds, the most popular kinds were aspergillus, which was isolated from swabs from the face and back, and penicillium isolated from the chest and smears from interdigital spaces of hands. one participant was found to be colonized by one species (candida lusitaniae) at all test sites on the skin. fungi were usually isolated from the interdigital spaces of fingers (n = 26, 43%), and least often from the back (n = 12, 18% of participants; fig. 1). there were no differences in the frequency of isolation of fungi on the grounds of sex tab. 1 frequency of isolation of fungi from different locations on the skin depending on the gender of the test participants. gender group of fungi face chest back hands n % n % n % n % f lack of fungi 25 62.50 26 63.41 31 75.61 23 63.41 yeasts 6 15.00 6 14.63 5 12.20 13 14.63 molds 9 22.50 7 17.07 3 7.32 1 2.44 mixed 0 0.00 2 4.88 2 4.88 4 9.76 m lack of fungi 11 73.33 11 73.33 13 86.67 7 46.67 yeasts 3 20.00 0 0.00 1 6.67 2 13.33 molds 1 6.67 4 26.67 1 6.67 4 26.67 mixed 0 0.00 0 0.00 0 0.00 2 13.33 f+m lack of fungi 36 65.45 37 66.07 44 78.57 30 53.57 yeasts 9 16.36 6 10.71 6 10.71 15 26.79 molds 10 18.18 11 19.64 4 7.14 5 8.93 mixed 0 0.00 2 3.57 2 3.57 6 10.71 f – female; m – male; n – number of samples. one participant refused to take a swab from her face because of her makeup. 3 of 5© the author(s) 2016 published by polish botanical society acta mycol 50(2):1068 talaga and krzyściak / non-lipophilic mycobiota of skin in relation to isolation location (cochran–mantel–haenszel test, p = 0.8371). however, after grouping the results for both sexes, it was found that the incidence of fungi depending on location is significantly different (chi-square test, p = 0.01191). in order to find a relationship between the location and the isolated fungi, correspondence analysis was carried out, the results of which are shown in fig. 2. therefore, it can be concluded that the occurrence of yeasts is associated with interdigital spaces, while the face and chest are associated with a higher incidence of molds. the back turned out to be the location from which fungi were isolated least frequently. face chest back interdigital spaces 0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% 36 37 44 30 9 6 6 15 10 11 4 5 2 2 6 prevalence of fungal colonization on the skin surface mixed molds yeasts absence body sites p e rc e n ta g e fig. 1 the incidence of fungal colonization depending on the skin location. molds mixed yeasts absence face chest back interdigital fig. 2 correspondence analysis: the presence of fungi and location. the blue indicates the location; the red indicates fungal group. 4 of 5© the author(s) 2016 published by polish botanical society acta mycol 50(2):1068 talaga and krzyściak / non-lipophilic mycobiota of skin discussion there are no effective methods to assess the actual presence of fungi on the skin. culture methods have their limitations associated with the lack of providing all essential conditions for growth for each species of mycobiota in vitro. it appears that the solution to this problem could be the application of molecular techniques. however, interpretation of the occurrence of species based on the isolation and sequencing of dna in projects investigating the human microbiota is also problematic. the mere discovery of fungal dna does not mean that an entire cell is present. and even if a fungal cell is present, it is not possible to assume that it is alive. another very important problem is widespread contamination of laboratory equipment such as reagents, test tubes, swabs, etc. with microbial dna [8]. furthermore, interpretation of the results of cultures and the results of molecular studies causes a number of problems mainly due to the fact that occasional isolation or finding the presence of fungal dna do not mean that a particular species colonizes the skin, it could be transient. fungi on the skin may occur accidentally as a result of contamination with soil, plant remnants, or after exposure to water or air. many fungi, including molds, are anemochorous; therefore, their spores are commonly airborne and may passively fall on the skin. this is confirmed by our findings indicating a lower incidence of fungi on the covered skin of the back and the highest prevalence in interdigital spaces which are exposed not only to the atmosphere but also to tap water and all kinds of contamination being the origin or the breeding ground for fungi. research of the mycobiome, independent of culture, showed that, apart from the dna of malassezia, the most common was the dna of penicillium and aspergillus molds [3]. also in the present study, based on culturing methods, these two kinds of fungi were predominant. most fungi were isolated from interdigital swabs. in the report entitled environmental and hand hygiene of the polish people, published by dettol and children’s memorial health institute [2], fungi were isolated in women from 17 (15 yeasts and 2 molds) out of 636 samples and in men from 10 (9 yeasts and 1 mold) out of 550 samples. the study suggests that there are no differences in the incidence of yeasts between the sexes (chi-squared test with yates’ correction for continuity, p = 0.5004, own calculations). our study also failed to demonstrate a more frequent colonization of interdigital spaces by yeasts in women than in men; it was approximately 14% for each group (fisher’s exact test, p = 0.3064). in this respect, it is curious that there is a higher incidence of candidiasis of interdigital spaces of hands and candidiasis of fingernails in women [9]. saprotrophic yeasts have similar life requirements to the requirements of the pathogen called candida albicans. it is known that candida albicans is usually found on the skin. other candida species that are predominant there are c. tropicalis, c. parapsilosis and c. orthopsilosis [3]. following deliberate inoculation of hands, after 45 minutes, candida parapsilosis showed greater survivability (c. albicans was also cultured in an amount of around 2.5–5% initial inoculum) [10]. the question arises of whether an earlier colonization by other species of yeasts is a safeguard against or predisposes people to infections with c. albicans. the presence of c. albicans on the skin of hands is usually associated with poor hygiene. these fungi live in the gastrointestinal tract and mucous membranes of the reproductive organs, from where they can be easily transferred to one’s hands, particularly when using the restroom. as reported in studies of the american society for microbiology, women wash their hands more often (75–93%) than men (58–77%) after using the restroom. in great britain, this is only 32% of men and 64% of women. these figures are somehow contrary to a higher incidence of mycosis in women. it is possible that mere washing without the use of disinfectants could also give rise to colonization. as was shown by biedunkiewicz and schulz’s research, 66% of tap water samples contained fungi, of which the most numerous were yeasts (81.3% of the isolated fungi) [1]. moreover, it is known that detergents damage the protective lipid layer of the skin [11], which, together with maintaining skin’s moisture (also through creams and emollients), may favor colonization and, consequently, infection. the problem of the presence of fungi on healthy skin and associating colonization with potential development of infection needs further investigation. one must 5 of 5© the author(s) 2016 published by polish botanical society acta mycol 50(2):1068 talaga and krzyściak / non-lipophilic mycobiota of skin remember that the promoted molecular testing techniques do not allow us to assess the presence and viability of cells and merely finding the presence of a particular species, whether with the use of molecular methods or culturing, does not mean that the given species is a permanent component of skin mycobiota. conclusions ■ human skin locations most contaminated with fungi are interdigital spaces of hands. ■ interdigital spaces of hands are dominated by yeasts, whereas molds are prevalent on the face and chest. however, there were usually no fungi isolations from the back. ■ there is no difference in the incidence of fungi depending on gender. references 1. biedunkiewicz a, schulz ł. fungi of the genus exophiala in tap water. mikologia lekarska. 2012;19:23–26. 2. raport marki dettol i centrum zdrowia dziecka. higiena rąk i otoczenia polaków [internet]. 2007 [cited 2016 jan 21]. available from: http://pis.lodz.pl/data/other/hr_polacy.pdf 3. findley k, oh j, yang j, conlan s, deming c, meyer ja, et al., topographic diversity of fungal and bacterial communities in human skin. nature. 2013;498(7454):367–370. http:// dx.doi.org/10.1038/nature12171 4. hannigan gd, grice ea. microbial ecology of the skin in the era of metagenomics and molecular microbiology. cold spring harb perspect med. 2013;3(12):a015362. http:// dx.doi.org/10.1101/cshperspect.a015362 5. pawłowicz a, adamski z. flora dermatofitowa i oportunistyczna w zmianach grzybiczych dłoni, stóp oraz paznokci u pacjentów kliniki dermatologii am w poznaniu w latach 1984-1994. mikologia lekarska. 1994;2:95–100. 6. de hoog gs, guarrao j, gene j, figueras mj. atlas of clinical fungi. 2nd ed. centraalbureau voor schimmelcultures utreht. reus: universitat rovira i virgili; 2000. 7. r core team. a language and environment for statistical computing [internet]. vienna: r foundation for statistical computing; 2013 [cited 2016 jan 21]. available from: http:// www.r-project.org 8. salter s, cox mj, turek em, calus st, cookson ws, moffatt mf, et al. reagent and laboratory contamination can critically impact sequence-based microbiome analyses. bmc biol. 2014;12:87 http://dx.doi.org/10.1186/s12915-014-0087-z 9. kwaśniewska j, szefer e, jaskółowska a. niektóre aspekty grzybic skóry i paznokci na podstawie danych pacjentów zakładu diagnostyki i leczenia chorób pasożytniczych i grzybic uniwersytetu medycznego w łodzi. mikologia lekarska. 2012;19(1):17–22. 10. rangel-frausto ms, houston ak, bale mj, fu c. wenzel rp. an experimental model for study of candida survival and transmission in human volunteers. eur j clin microbiol infect dis. 1994;13(7):590–595. http://dx.doi.org/10.1007/bf01971311 11. okuda m, yoshiike t, ogawa h. detergent-induced epidermal barrier dysfunction and its prevention. j dermatol sci. 2002;30(3):173–179. http://dx.doi.org/10.1016/ s0923-1811(02)00106-8 http://pis.lodz.pl/data/other/hr_polacy.pdf http://dx.doi.org/10.1038/nature12171 http://dx.doi.org/10.1038/nature12171 http://dx.doi.org/10.1101/cshperspect.a015362 http://dx.doi.org/10.1101/cshperspect.a015362 http://www.r-project.org http://www.r-project.org http://dx.doi.org/10.1186/s12915-014-0087-z http://dx.doi.org/10.1007/bf01971311 http://dx.doi.org/10.1016/s0923-1811(02)00106-8 http://dx.doi.org/10.1016/s0923-1811(02)00106-8 abstract introduction material and methods results discussion conclusions references 2016-01-26t13:40:10+0000 piotr otręba 2014-11-20t23:15:07+0000 polish botanical society 2014-11-20t23:21:21+0000 polish botanical society 2014-11-20t23:18:12+0000 polish botanical society 2014-11-20t23:15:21+0000 polish botanical society 2014-11-20t23:20:04+0000 polish botanical society 2014-11-20t23:16:37+0000 polish botanical society 18th congress of european mycologists: conference report acta mycologica article id: 55214 doi: 10.5586/am.55214 publication history received: 2020-11-15 accepted: 2020-11-16 published: 2021-03-01 handling editor wojciech pusz; wrocław university of environmental and life sciences, poland; https://orcid.org/0000-00031531-2739 authors, contributions all authors contributed to the writing of this report. funding the event was cofinanced by the polish ministry of science and higher education under its objective to disseminate science (contract no. 964/p-dun/2018) and by the following sponsors: the european mycological association, cybertruffle, analityk, biomaxima, eurx molecular biology products, genomed and precoptic companies. competing interests no competing interests have been declared. copyright notice © the author(s) 2020. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. report 18th congress of european mycologists: conference report julia pawłowska 1,2*, magdalena frąc 1,3, izabela kałucka 1,4, małgorzata ruszkiewicz-michalska 1,4,5, sylwia różalska 1,4, marta wrzosek 1,6 1polish mycological society, poland 2institute of evolutionary biology, faculty of biology, biological and chemical research center, university of warsaw, poland 3institute of agrophysics, polish academy of sciences, poland 4faculty of biology and environmental protection, university of lodz, poland 5institute for agricultural and forest environment, polish academy of sciences, poland 6botanic garden, faculty of biology, university of warsaw, poland *to whom correspondence should be addressed. email: julia.z.pawlowska@uw.edu.pl the 18th congress of european mycologists was held between september 16 and 21, 2019 in warsaw and białowieża, poland. it gathered more than 250 participants from 52 countries from all over the world who presented 77 talks and 176 posters (hawksworth, 2019; pawłowska et al., 2019). during the first 3 days, mycologists were meeting in old library at the university of warsaw and they could participate in two offered and six thematic sessions: “from genome to function” (led by ekaterina shelest, germany), “taxonomy and systematics” (led by jos houbraken, the netherlands), “fungi in biotechnology” (led by katarzyna turnau, poland), “fungal interactions” (led by martin bidartondo, united kingdom), “medical mycology” (led by michaela lackner, austria), and “fungal diversity” (led by carrie andrew, sweden). four thematic sessions were organized in białowieża: “fungi in primeval forests and other natural habitats” (led by anders dahlberg, sweden), “hypogeous mycorrhizal fungi” (led by giovanni pacioni, italy), “fungal conservation” (led by susana c. gonçalves, portugal), and “data session” (led by dmitry schigel, denmark). additionally, two workshops were organized in the białowieża part of the congress: “global fungal red-listing” (by david minter, united kingdom) and “biology of polypores” (by dmitry schigel, denmark). an acta mycologica / 2020 / volume 55 / issue 2 / article 55214 publisher: polish botanical society 1 https://doi.org/10.5586/am.55214 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0003-4914-5182 https://orcid.org/0000-0001-9437-3139 https://orcid.org/0000-0002-7264-5735 https://orcid.org/0000-0001-8901-0552 https://orcid.org/0000-0003-1695-5154 https://orcid.org/0000-0002-5871-5020 mailto:julia.z.pawlowska@uw.edu.pl pawłowska et al. / 18th cem report open european council for the conservation of fungi (eccf) discussion forum entitled “fungal conservation across borders in europe (and beyond) for the next 4 years – what role for the eccf?” was also held. the scientific program was also honored by keynote lectures of eminent mycologists: annegret kohler (france), david hawksworth (united kingdom), geoffrey gadd (united kingdom), duur k. aanen (the netherlands), dominik begerow (germany), marc-andré selosse (france), bogdan jaroszewicz (poland), and lynne boddy (united kingdom). at a general meeting of the european mycological association, a new board was elected, with izabela kałucka (poland) as president, mitko karadelev (macedonia) as vice-president, katerina rusevska (macedonia) as secretary, eske de crop (belgium) as treasurer, vijai kumar gupta (estonia) as membership secretary, tatiana semenova-nelsen (usa) as publicity & social media officer, paulo de oliveira (portugal) as ema webmaster, and susana c. gonçalves (portugal) as conservation officer and chair of the eccf. the outgoing president, david minter, was thanked for all he had done for the ema during his tenure. during the closing ceremony, it was agreed that the next, the 19th congress of european mycologists will be organized in turin, italy, in 2023. as the motto of the congress was “fungi in nature and culture,” a rich program of mycology related, artistic events was proposed, e.g., a photo exhibition “fungi – master sculptures of nature,” “dystopia” – spatial composition by hélène soulier and ewa rudnicka, “anomalium” – graphics exhibition of agnieszka zdziabek, “broken links” – performance of maria subczyńska. additionally, two mushroom exhibitions in warsaw and the 25th exhibition of fungi of the białowieża forest were also organized. on tuesday evening, the public lecture “mycology: a recent weapon in the forensic armory” was given in the copernicus science centre by patricia wiltshire (united kingdom). finally, common mushroom hunting and inaturalist.org bioblitz project were also organized, as a follow up of the gbif presentation stressing the importance of sharing data on fungal occurrence. in less than 2 weeks, 750 observations and 217 species of fungi from białowieża forest (poland) were recorded. some collections were also taken for further identification. all these data will be processed and prepared for publication in the acta mycologica journal. this will be the first paper opening a new chapter in the journal’s history. acta mycologica, as one of the world’s first mycological journals, has decided to join the gbif initiative to share data on fungal distribution. we hope that such a decision will significantly increase the representativeness of polish mycological data in international databases and will become a measurable and globally visible effect of the recent congress. acknowledgments the event was organized jointly by the polish mycological society and the european mycological association and it was co-organized by the faculty of biology and botanical garden – university of warsaw; faculty of biology and environmental protection – university of lodz; institute of agrophysics, polish academy of sciences; forest research institute; białowieża national park; institute for agricultural and forest environment, polish academy of sciences; faculty of forestry of the białystok university of technology in hajnówka; and warsaw convention bureau. the congress was held under honorary patronages of the minister of science and higher education of the republic of poland, marshal of the mazowieckie voivodeship, marshal of the podlaskie voivodeship, mayor of warsaw, director of białowieża national park, rector of the university of warsaw, president of the polish academy of sciences, and general director of the state forests. references hawksworth, d. l. (2019). myconews 2019: editorials, news, reports, awards, personalia, book news, and correspondence. ima fungus, 10(1), article 23. https://doi.org/10.1186/s43008-019-0024-4 pawłowska, j., frąc, m., kałucka, i., ruszkiewicz-michalska, m., różalska, s., & wrzosek, m. (2019). the xviii congress of european mycologists: conference report. journal of fungi, 5, article 110. https://doi.org/10.3390/jof5040110 acta mycologica / 2020 / volume 55 / issue 2 / article 55214 publisher: polish botanical society 2 https://doi.org/10.1186/s43008-019-0024-4 https://doi.org/10.3390/jof5040110 acknowledgement references new and interesting lichen records from northeastern poland 1 of 5published by polish botanical society acta mycologica short communication new and interesting lichen records from northeastern poland dariusz kubiak*, ewa sucharzewska department of mycology, university of warmia and mazury in olsztyn, oczapowskiego 1a, 10-957 olsztyn, poland * corresponding author. email: darkub@uwm.edu.pl abstract details are given of the occurrence of three rarely reported and poorly known lichen species from poland area. brief taxonomic, distributional, and ecological notes of agonimia flabelliformis, bacidia pycnidiata, and vezdaea aestivalis have been provided. agonimia flabelliformis have been reported for the first time from the northeastern part of poland and b. pycnidiata from the northern part of the country. vezdaea aestivalis has been rediscovered in northeastern poland, nearly 150 years after its first and only recording in the region. keywords lichenized fungi; rare species, distribution; new localities; ne poland introduction the northeastern part of poland is a relatively well-preserved natural environment with a cultural landscape. compared to other lowland areas of the country, this region is characterized by high species diversity of different groups of organisms, including lichens [1]. at the supra-regional scale, very important refuges for the diversity of lichen species are the large forest complexes of this region. they provide important habitats for a diverse group of stenotopic forest lichens, rarely observed or even absent in other parts of the country [2–5]. however, the state of scientific knowledge on diversity of lichen species in this region is still unsatisfactory. over the last 10 years, studies on the lichen biota in forest ecosystems in this area has been intensified, and resulted in finding of many interesting taxa, including some that are new to the country [6–8]. in our paper, new localities of three noteworthy lichen species, occurring in poland in very limited amounts, are presented. our data will contribute to a better understanding of the habitat requirements and population dynamics of these species. material and methods lichen specimens described in the present paper was collected in years 2010–2016 as a result of different research projects, realized in the best preserved forest complexes of the pojezierze mazurskie lakeland [9]. the collected specimens were evaluated using standard methods [9]. identification of sterile specimens was supported by tlc analyses of secondary metabolites [10]. the distribution of the taxa examined is given in the atpol grid square system [11], modified by cieśliński and fałtynowicz [12]. the collected material is deposited in the herbarium of the department of mycology uwm in olsztyn (oltc). abbreviations: fd – forest division, fs – forest section, nr – nature reserve. doi: 10.5586/am.1073 publication history received: 2016-04-25 accepted: 2016-06-16 published: 2016-06-24 handling editor tomasz leski, institute of dendrology, polish academy of sciences, poland authors’ contributions dk, es: field research, writing the manuscript; dk: idea of the study, determination of the specimens, preparation of the distribution maps funding the study was funded by the faculty of biology and biotechnology, university of warmia and mazury in olsztyn. competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation kubiak d, sucharzewska e. new and interesting lichen records from northeastern poland. acta mycol. 2016;51(1):1073. http:// dx.doi.org/10.5586/am.1073 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:darkub%40uwm.edu.pl?subject=new%20and%20interesting%20lichen%20records%20from%20northeastern%20poland http://dx.doi.org/10.5586/am.1073 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1073 http://dx.doi.org/10.5586/am.1073 2 of 5© the author(s) 2016 published by polish botanical society acta mycol 51(1):1073 kubiak and sucharzewska / new and interesting lichen records results and discussion agonimia flabelliformis halda, czarnota & guzow-krzemińska this recently described species [13] is distinguished by distinctly raised, coralloid to palmate (flabelliform) thallus (bright green to pale brown-green) and globose, smooth perithecia (pale brown to dull grey-brown) with eight-spored asci. agonimia flabelliformis is most similar to a. allobata due to comparable perithecia, however, the latter species has usually warted to subsquamulouse thallus [13]. agonimia flabelliformis prefers humid, shaded, mossy places within deciduous forest ecosystems, where it usually grows epibryophytically over bark (at the base of trees and their roots) or occasionally on soil, rocks, wood, or plant debris [13–17]. at the new collection sites, the species was growing on corticolous bryophytes (hypnum cupressiforme) and directly on bark at the base of mature oak trees. so far a. flabelliformis is known only from europe, where it has a wide range of occurrence. it has been found in great britain, germany, the czech republic, slovakia, poland, lithuania, and russia [13,14,18,19]. in poland, it has been noted in only three localities (fig. 1), in southern [14], central [15], and northwestern parts of the country [20]. the specimens examined. be-22: pojezierze olsztyńskie lakeland, wichrowo fd, fs no. 403h, 54°02'04.2" n, 20°23'36.9" e, on quercus robur, 20 may 2010, leg. d. kubiak (oltc l-3567). be-62: pojezierze olsztyńskie lakeland, nowe ramuki fd, fs no. 234b, 53°40'35.0" n, 20°25'54.6" e, on q. robur, 4 oct. 2014, leg. d. kubiak (oltc l-3624). bacidia pycnidiata czarnota & coppins bacidia pycnidiata is a very characteristic member of the genus due to its long-necked whitish or cream pycnidia (usually immersed within green, minutely granular thallus), and more or less straight macroconidia [21]. very similar pycnidia appear occasionally in fellhanera subtilis (vězda) diederich & sérus., however, conidia in the latter species are short and pyriform, and its thallus is smooth to scurfy. the species shows preference for moderately shaded, old-growth or undisturbed broad-leaved forests, where it grows on the mossy bark of deciduous trees, and very rarely on mossy soil or limestone [22]. in poland, it has been noted in forests on the trunks of fraxinus excelsior and quercus sp. [23,24] as well as on the moribund thallus of peltigera didactyla in dry sandy habitats [25]. some data from strongly industrialized regions of the czech republic and southern poland suggest that b. pycnidiata could be a synanthropic species. the species usually occurs there in anthropogenic habitats, where prefers humid niches [21,25,26]. at the new collection site, the species was growing in an anamorphic state only on corticolous bryophytes at the base of a mature oak tree within old-growth oak forest. bacidia pycnidiata is found mainly in central europe. it has been reported in belgium, the czech republic, slovakia, poland, lithuania, estonia, finland, ukraine, and russia [22,27]. in poland, it has so far been reported at five localities (fig. 2) in the uplands and the mountainous regions in the southern part of the country [21,23,25]. the specimen examined. be-63: pojezierze olsztyńskie lakeland, nowe ramuki fd, fs no. 101f, 53°40'37" n, 20°31'48"e, on q. robur, 12 mar. 2016, leg. d. kubiak (oltcl-3627). a b c d e f g a b c d e f g o 52 o 54 o 50 o 16 o 20 o 24 1 2 fig. 1 distribution of agonimia flabelliformis in poland. 1 – known localities; 2 – new localities. 3 of 5© the author(s) 2016 published by polish botanical society acta mycol 51(1):1073 kubiak and sucharzewska / new and interesting lichen records vezdaea aestivalis (ohlert) tscherm.-woess & poelt this is the largest and most conspicuous species of the genus vezdaea [28], distinguished from other species by relatively large apothecia (up to 1 mm in diam.) and certain anatomical characteristics [e.g., paraphyses closely clasp the asci, 1(–3)-septate and verrucose spores, goniocysts with short conical spines] [29]. vezdaea aestivalis superficially resembles the micarea species, but shortlived convex tomentose apothecia without exciple and hypothecium are diagnostic characteristics [30]. the species has been found in europe (belarus, the british isles, the czech republic, denmark, estonia, germany, lithuania, poland, romania, russia, slovakia, spain) and australia [28,31,32]. the first report of v. aestivalis within polish territory (as lecidea aestivalis ohlert) comes from the nineteenth century [33], but for more than one hundred years there were no new sightings recorded. since the last report from alstrup and olech [34], many new localities with it have been published. vezdaea aestivalis is currently known to be in dozens of localities in the southern part of the country (upland and mountainous areas), but only one report has come from the northern part [35–38]. the species has been studied in detail and mapped in poland by czarnota and kiszka [35] and czarnota and kukwa [29]. vezdaea astivalis has a huge ecological plasticity. it is found on mosses, terricolous lichens and plant remains amongst or on rocks (mainly calcareous), in limestone grassland, on walls, rubble, building ruins, and waste ground. it has also been found in shaded or moist sites, and amongst mosses on trees with base-rich bark [35]. in poland, the species has been noted both in habitats strongly influenced by human activities, e.g., abandoned zinc-lead mines and piles of old artificial fertilizers [37,38], and undisturbed areas, e.g., natural forests [35]. the specimen presented in this paper is an example of the latter situation; it was found at the base of a mature oak between corticolous bryophytes (pylaisia polyantha), in an oak-hornbeam forest. the specimen examined. be-62: pojezierze olsztyńskie lakeland, nowe ramuki fd, fs no. 784g, 53°36'49" n, 20°27'36" e, on a trunk of q. robur, 3 oct. 2015, leg. d. kubiak (oltc l-3625). acknowledgments the authors are very grateful to prof. jakub sawicki (university of warmia and mazury in olsztyn, poland) for the determination of moss species, and to the editor and anonymous reviewers for their useful comments and advices. references 1. cieśliński s. atlas rozmieszczenia porostów (lichenes) w polsce północno-wschodniej. warszawa: białowieska stacja geobotaniczna uw; 2003. (phytocoenosis – supplementum cartographiae geobotanicae; vol 15). a b c d e f g a b c d e f g o 52 o 54 o 50 o 16 o 20 o 24 1 2 fig. 2 distribution of bacidia pycnidiata in poland. 1 – known localities; 2 – new locality. a b c d e f g a b c d e f g o 52 o 54 o 50 o 16 o 20 o 24 1 2 fig. 3 distribution of vezdaea aestivalis in poland. 1 – known localities; 2 – new locality. 4 of 5© the author(s) 2016 published by polish botanical society acta mycol 51(1):1073 kubiak and sucharzewska / new and interesting lichen records 2. czyżewska k, cieśliński s. porosty – wskaźniki niżowych lasów puszczańskich w polsce. łódź: polskie towarzystwo botaniczne; 2003. 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2004. p. 107–110. 36. czyżewska k, hachułka m, łubek a, zaniewski p. distribution of some lichenicolous fungi in poland. ii. acta mycol. 2008:3(2):193–206. http://dx.doi.org/10.5586/am.2008.024 37. bielczyk u, jędrzejczyk-korycińska m, kiszka j. lichens of abandoned zinc-lead mines. acta mycol. 2009;44(2):139–149. http://dx.doi.org/10.5586/am.2009.012 38. kościelniak r, betleja l. porosty na nietypowych podłożach antropogenicznych w bieszcza dzkim parku narodowym. roczniki bieszczadzkie. 2013;21:35–41. http://dx.doi.org/10.2478/v10111-012-0003-7 http://dx.doi.org/10.2478/botcro-2013-0025 http://dx.doi.org/10.13158/heia.27.2.2014.257 http://dx.doi.org/10.13158/heia.27.2.2014.257 http://dx.doi.org/10.1017/s0024282987000136 http://dx.doi.org/10.1515/botlit-2015-0007 http://dx.doi.org/10.5586/am.2008.024 http://dx.doi.org/10.5586/am.2009.012 abstract introduction material and methods results and discussion agonimia flabelliformis halda, czarnota & guzow-krzemińska bacidia pycnidiata czarnota & coppins vezdaea aestivalis (ohlert) tscherm.-woess & poelt acknowledgments references 2016-06-24t10:01:58+0100 piotr otręba notes on some phytopythium and pythium species occurring in oak forests in southern poland 1 of 10published by polish botanical society acta mycologica original research paper notes on some phytopythium and pythium species occurring in oak forests in southern poland robert jankowiak1*, hanna stępniewska1, piotr bilański2 1 department of forest pathology, mycology and tree physiology, university of agriculture in krakow, al. 29 listopada 46, 31-425 cracow, poland 2 department of forest protection, entomology and forest climatology, university of agriculture in krakow, al. 29 listopada 46, 31-425 cracow, poland * corresponding author. email: rljankow@cyf-kr.edu.pl abstract phytopythium and pythium species are known to be soil-born oomycete pathogens of forest trees in europe. little is known, however, about the presence of these micro-organisms in polish oak forests. during the period 2007–2009 a comprehensive study of phytophthora species in soils of oak forests in southern poland was conducted using baiting technique. in this study, baits were also colonized by oomycete resembling pythium species. based on morphological characteristics and the its sequences comparisons, 10 species of phytopythium and pythium were isolated from the soil-root samples, including three putative new species belonging to the genus of phytopythium. the most commonly encountered pythium species was pythium anandrum. the present study demonstrates for the first time that phytopythium citrinum and pythium diclinum can also act as soil-borne organisms in oak forests. in addition, these species were reported for the first time in poland. keywords quercus robur; oomycetes; soil; phytopythium; pythium this issue of acta mycologica is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. introduction the genus pythium pringsh. including “fungus-like organisms” or “pseudo-fungi” is placed in the kingdom chromista [1] or kingdom straminipila [2]. the species of pythium are cosmopolitan, widely distributed throughout the world, and occupy several diverse ecological niches. some species of pythium are known as pathogens of various plants, including forest and fruit trees; they lead to rot of fruit, rot of roots, and stems, and preor postemergence damping-off. for example pythium undulatum h.e. petersen, causes root rot of abies procera rehder and pseudotsuga menziesii (mirb.) franco in northern germany [3]. the genus pythium is characterized by filamentous or globose sporangia with zoospores develop in a vesicle, oospores formed in smooth or ornamented oogonia with paragynous or hypogynous antheridia [4]. many reports have shown that pythium is composed of few morphological groups, whereas recent molecular analyses have shown that the genus pythium is a polyphyletic group that includes several monophyletic groups [5,6]. in a recent study, uzuhashi et al. [7] restricted the genus pythium to those species with filamentous sporangia and created four new genera to accommodate species with non filamentous sporangia: ovatisporangium, elongisporangium, globisporangium and pilasporangium. in the same year bala [8] proposed a new genus doi: 10.5586/am.1052 publication history received: 2014-11-28 accepted: 2015-05-18 published: 2015-08-05 handling editor maria rudawska, institute of dendrology of the polish academy of sciences, poland authors’ contributions rj: writing the paper, identification of isolates and analyzing the data; hs: collecting of samples and isolates, identification of isolates; pb: analyzing the data funding research was carried out under the theme no. ds 3414/ zflm i fd funded by a grant to study awarded by the polish ministry of science and higher education. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation jankowiak r, stępniewska h, bilański p. notes on some phytopythium and pythium species occurring in oak forests in southern poland. acta mycol. 2015;50(1):1052. http://dx.doi. org/10.5586/am.1052 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:rljankow%40cyf-kr.edu.pl?subject=notes%20on%20some%20phytopythium%20and%20pythium%20species%20occurring%20in%20oak%20forests%20in%20southern%20poland http://dx.doi.org/10.5586/am.1052 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1052 http://dx.doi.org/10.5586/am.1052 2 of 10© the author(s) 2015 published by polish botanical society acta mycol 50(1):1052 jankowiak et al. / pythium species occurring in oak forests phytopythium for those species with globose to ovoid, often papillate and internally proliferating sporangia. recently, de cock et al. [9] provided molecular-based evidence that members of pythium clade k as described by lévesque and de cock [5] belong to the phytopythium genus. while recognizing the genus status of remaining species of pythium clades (a–j) is still unclear we prefer to use to the definition of pythium sensu lévesque and de cook [5]. little is known about the occurrence of phytopythium and pythium species in europe, particularly in forest soils. there were only four reports about isolation of pythium spp. from soils under oak forests [10–13]. these studies reported occurrence of p. undulatum, p. anandrum drechsler, p. aphanidermatum (edson) fitzp., p. irregulare buisman, p. middletonii sparrow, p. rostratum butler and p. intermedium de bary in oak stands in austria, germany, turkey and sweden. of all pythium species recorded in these studies, p. anandrum and p. undulatum are the most commonly reported and are known as fine root pathogens which may affect the health of oak trees in europe [9–13]. recently, pythium sterilum, p. spiculum b. paul belbahri & lefort, and two phytopythium species, phytopythium citrinum-like (b. paul) abad, de cock, bala, robideau, lodhi & lévesque and phytopythium mercuriale (belbahri, b. paul & lefort) abad, de cock, bala, robideau, lodhi & lévesque have been associated with soil in declining oak stands in poland [14]. however, information about the oak-associated pythium and phytopythium species is still very limited. during the period 2007–2009 a comprehensive study of phytophthora species in soil samples collected from oak forests in southern poland was conducted using baiting (oak leaves) and selective agar medium technique [15]. in this study, baits were also colonized by other oomycetes, particularly by pythium species. for this reason, our objective was to identify the pythium isolates obtained during phytophthora detecting in oak forests in southern poland. material and methods study sites the survey was conducted during may–june and september–october 2007–2009 in 29 pedunculate oak (quercus robur l.) stands in the southern part of poland. sampling sites were selected from areas characterized by different tree health status and site conditions. the characteristics of the study sites are given in tab. 1. sampling and isolation methods in each stand 6 mature trees (tree age >50 years), which were considered to be representative for the health status of the stand, were chosen. the crown status of these trees was assessed according to balcì and halmschlager [11]. three soil samples were taken with fine roots (soil-root monoliths without the organic part ca. 25 × 25 × 25 cm), 100–150 cm from the trunk and spaced in three directions around the stem base of each tree. soil from three monoliths was mixed and a sub-sample of this mixture was put in a plastic bag and transported to the laboratory. isolation was performed using the oak leaf baiting method described by jung et al. [10,16]. each sample was mixed thoroughly, then two 200 ml subsamples were flooded with 400 ml of distilled water in a plastic boxes (18 × 11 × 7 cm), and baited by floating 2to 6-day-old q. robur leaves (10–15 leaves per each box) at room temperature. after 3–6 days, discolored leaves were taken to phytophthora spp. isolation. for this purpose, leaves were washed under tap water, dried on filter paper, cut into small pieces (2 × 2 mm) and placed on parpnh medium (v8 juice agar amended with 10 mg/l pimaricin, 200 mg/l ampicillin, 10 mg/l rifampicin, 25 mg/l pcnb, 50 mg/l nystatin and 50 mg/l hymexazol). emerging cultures were purified by transferring small pieces of mycelium from individual colonies to fresh v8 juice agar (100 ml/l vega’s juice (tymbark®, poland), 900 ml/l distilled water, 15 g/l agar, 3 g/l caco3). 3 of 10© the author(s) 2015 published by polish botanical society acta mycol 50(1):1052 jankowiak et al. / pythium species occurring in oak forests tab. 1 characteristics of the study sites (in bold the sites where phytopythium/pythium spp. were found). geographic coordinates sites no. site longitude latitude forest type1 tree age soil texture2 soil moisture3 soil ph4 cacl2 crown status5 1 ispina 1 20.362721° 50.096364° q 130 silty clay loam m 3.69 sd 2 ispina 2 20.359564° 50.081254° d 165 silty clay loam m 3.75 sd 3 wola batorska 1 20.269651° 50.032460° p 120 sand m 2.83 sd 4 wola batorska 2 20.249682° 50.027453° q 195 silty clay loam m 3.57 d 5 stanisławice 1 20.335946° 49.991910° p 65 sand mm 3.55 sd 6 stanisławice 2 20.324853° 49.932567° p 125 sand m 2.92 sd 7 krzyszkowice 20.016746° 50.002342° d 150 silt loam mm 3.75 d 8 ostrowy tuszowskie 21.643837° 50.326383° q 135 sand mm 3.61 sd 9 przyborów 1 15°45'06" 51°47'27" q 150 sandy loam pfwt 4.82 d 10 przyborów 2 15°45'07" 51°46'58" q 140 sandy loam pfwt 5.39 d 11 babice 18°16'03" 50°08'01" q 120 sandy loam mm 3.77 sd 12 maleniska 22°19'59" 50°16'00" d 70 sand mm 3.32 h 13 przedbórz 19°54'17" 51°05'41" p 60 sand md 3.46 sd 14 rączna 19°44'52" 50°00'26" d 70 sand md 3.51 h 15 radymno 23°04'00" 49°56'60" p 115 loamy sand mm 4.22 d 16 pomorsko 15°28'47" 52°02'44" q 150 sand pfwt 5.20 h 17 piskorowice 22°34'40" 50°14'04" p 80 loamy sand mm 3.75 sd 18 pociękarb 18°04'44" 50°18'58" q 150 sandy loam mm 3.14 sd 19 bierdzany 18°09'10" 50°48'19" d 80 loamy sand mm 3.33 h 20 rzędzów 18°08'22" 50°44'18" d 50 sand mm 3.52 sd 21 chełmiec 16°04'58" 51°02'13" q 100 silt loam mm 3.6 h 22 moszna 17°46'25" 50°25'31" d 90 loamy sand mm 3.32 sd 23 leśniki 15°50'22" 52°07'39" p 105 sand md 5.94 sd 24 laski 15°48'32" 52°10'27" d 135 sandy loam mm 3.29 d 25 kręcko 15°04'01" 52°10'27" d 125 sand mm 5.18 sd 26 zwierzyniec 18°45'31" 50°55'30" q 140 sandy loam mm 3.99 h 27 głubczyce 17°38'39" 50°09'50" q 135 silt loam m 3.68 h 28 mikolin 17°40'48" 50°47'41" q 120 silt loam m 3.89 sd 29 czyżowice 18°22'39" 49°59'33" q 90 silty clay loam mm 3.58 sd 1 q – pure quercus robur stand; p – mixture with pinus sylvestris; d – mixture with other deciduous species. 2 soils texture according to usda (united states department of agriculture). 3 m – moist; mm – moderately moist; d – dry; md – moderately dry; pfwt – periodically fluctuating water table. 4 the ph was measured with a glass electrode in a 0.01 m cacl2 suspension and in deionized h2o. 5 h – healthy (class 1 according to balcì and hamschlager [10]); sd – slightly damaged (class 2); d – declining (class 3 and 4). 4 of 10© the author(s) 2015 published by polish botanical society acta mycol 50(1):1052 jankowiak et al. / pythium species occurring in oak forests identification of pythium and phylogenetic analysis all pythium cultures were grouped according to morphological characters using a nikon eclipse 50i microscope (nicon® corporation, tokyo, japan) and an invenio 5s digital camera (deltapix®, maalov, denmark) with coolview 1.6.0 software (precoptic®, warsaw, poland). pythium structures and colony characteristics were compared with the descriptions of species given in the literature [17]. the morphology of colonies was described from 7-day-old colonies growing on carrot (ca), cornmeal agar (cma), potato-dextrose agar (pda), malt agar (mea) and v8 agar medium. sporangia were obtained by flooding agar discs taken from growing margins of 7-dayold colonies with unsterile soil extract (ratio 1:10). from each morphological group, isolates were selected for dna sequencing and were deposited in the fungal culture collection of the department of forest pathology, mycology and tree physiology, hugo kołłątaj university of agriculture, cracow, poland. genomic dna from 18 strains (tab. 2) was isolated using archivepure dna yeast/gram-positive bacteria kit (5 prime, inc. gaithersburg, md) with modified time of incubation with lytic enzyme solution (2 h, 37°c) and cell lysis solution (4 h, 64°c). nuclear its rdna region internal transcribed spacers (its1 and its2) and 5.8s subunits were amplified with primer set its5/its4 [18]. the reaction mixtures and conventional pcr protocols were the same as in hubka and kolarik [19]. custom purification of pcr amplicons and sequencing was conducted at macrogen inc. (seoul, south korea) using the same primers. tab. 2 cultures used in this study and genbank accession numbers for sequences. taxon isolate number1 site accession no. closest match in blast accession of match identity (%) phytopythium citrinum 121hr09 zwierzyniec kc602480 phytopythium citrinum hq 643379 100.0 143hr09 zwierzyniec kc602481 phytopythium citrinum hq 643379 100.0 35hr09 zwierzyniec kc602482 phytopythium citrinum hq 643379 100.0 48hr09 zwierzyniec kc602483 phytopythium citrinum hq 643379 100.0 99hr09 głubczyce kc602484 phytopythium citrinum hq 643379 100.0 phytopythium cf. citrinum a 444hr08 przyborów 2 kc602485 pythium sp. gd33b ef152505 99.7 447hr08 przyborów 2 kc602486 pythium sp. gd33b ef152505 99.7 phytopythium cf. citrinum b 428hr08 przyborów 2 kc602487 pythium sp. gd33b ef152505 100.0 520hr08 przyborów 1 kc602488 pythium sp. gd33b ef152505 100.0 phytopythium sp. 1 68hr09 zwierzyniec kc602492 pythium sp. b57 jn863966 95.9 phytopythium sp. 2 451hr08 przyborów 2 kc602493 pythium sp. pv so7 eu669081.1 94.0 phytopythium sp. 3 474hr08 przyborów 1 kc602494 phytopythium sp. mab-2011e ab690623.1 97.5 pythium anandrum 304hr07 ispina 2 kc602489 pythium anandrum hq643435 100.0 305hr07 ispina 2 kc602490 pythium anandrum hq643435 100.0 pythium diclinum 27hr09 laski kc602491 pythium diclinum jq898459 100.0 pythium intermedium 2hr09 kręcko kc602495 pythium intermedium ay083936 100.0 pythium undulatum 719hr08 radymno kc602496 pythium undulatum eu240049 100.0 725hr08 radymno kc602497 pythium undulatum eu240049 100.0 1 kfl: fungal culture collection, department of forest pathology, mycology and tree physiology, hugo kołłątaj university of agriculture, cracow, poland. 5 of 10© the author(s) 2015 published by polish botanical society acta mycol 50(1):1052 jankowiak et al. / pythium species occurring in oak forests sequences were compared with the data from genbank using a blast search. all sequences were aligned online with mafft v6 [20], using the e-ins-i option with a gap-opening penalty of 1.53 and an offset value of 0.00. datasets were analyzed using maximum likelihood (ml) and bayesian inference (bi). for the ml and bayesian analyses, the best-fit substitution models for each data set were established using the corrected akaike information criterion (aicc) in jmodeltest 0.1.1 [21]. the selected model for the its was gtr+i+g. ml searches were conducted in phyml 3.0 [22], via the montpelier online server (http://www.atgc-montpellier.fr/phyml/) with 1000 bootstrap replicates. bi analyses based on a markov chain monte carlo (mcmc) were performed with mrbayes v3.1.2 [23]. the mcmc chains were run for 10 million generations using the best fitting model. trees were sampled every 100 generations, resulting in 100 000 trees from both runs. the burn-in value for each dataset was determined in tracer v1.4.1 [24]. all sequences generated in this study were deposited in the ncbi genbank (tab. 2) and are presented in the phylogenetic tree (fig. 1, fig. 2). pythium clades were designated according to lévesque and de cock [5]. results morphological investigation showed that ten groups producing sporangia and sexual structures in culture were collected. the its data confirmed that these groups represented taxa belonging to the phylogenetically related genera, phytopythium and pythium. groups with affinity to the genus phytopythium represented one known species (phytopythium citrinum) and five unknown species named here as phytopythium cf. citrinum a, phytopythium cf. citrinum b, phytopythium sp. 1, phytopythium sp. 2 and phytopythium sp. 3. in turn, the genus pythium was represented by four species including pythium anandrum, p. diclinum tokun., p. intermedium and p. undulatum (fig. 1, fig. 2). polygenetic analyses of the its sequences of pythium isolates placed them in three clades (b, f, h) within pythium (in lévesque, de cock [5]). phytopythium cf. citrinum a and phytopythium cf. citrinum b in phylogenetic analyses were most closely related to isolate pythium sp. gd33b, while the three unknown species were most closely related to different species in the genus phytopythium (fig. 2, tab. 2). their identity and morphological features are still under investigation and descriptions of these taxa will be provided in a later publication. ten different phytopythium and pythium species were isolated from rhizosphere soil in 8 of the 29 oak stands. among them, pythium anandrum was most frequently recorded (4.0%) and showed the widest geographical distribution (4 stands). phytopythium citrinum, phytopythium cf. citrinum b, phytopythium sp. 1 and phytopythium sp. 3 were found in two stands. on average, the isolation frequency of phytopythium citrinum, phytopythium cf. citrinum b, phytopythium sp. 1 and phytopythium sp. 3, in the oaks forests was 2.3%, 1.7%, 2.3% and 2.9%, respectively. other species were sporadically isolated from rhizosphere soil (tab. 3). among the six identified soil textures, phytopythium spp. and pythium spp. were most frequently isolated from soil developed from sandy loam. these organisms occurred on sites with a mean soil ph range (cacl2) ranging from 3.29 to 5.39, and were most often isolated from soil samples taken from declining trees (14 out of 36) and from pure q. robur stands (4 out of 29; tab. 1). discussion this is the first extensive report demonstrating the presence of phytopythium and pythium species in oak forests in poland. based on morphological characteristics and its rdna sequence analysis, 10 species of phytopythium spp. and pythium spp. were isolated from the soil-root samples, including three putative new species. the most commonly encountered pythium species was pythium anandrum. the present study demonstrates for the first time that phytopythium citrinum and pythium diclinum can http://www.atgc-montpellier.fr/phyml/ 6 of 10© the author(s) 2015 published by polish botanical society acta mycol 50(1):1052 jankowiak et al. / pythium species occurring in oak forests also act as soil-borne oomycete species in oak forests. in addition, these species were reported for the first time in poland. this study has shown that the assemblage of phytopythium/pythium spp. occurring in the rhizosphere soil of oak forests in poland was quite diverse. the occurrence of phytophthora ramorum hq643339 phytophthora lateralis hq875390 phytopythium carbonicum hq643373 phytopythium montanum hq643391 pythium violae hq643968 pythium splendens jn585826 pythium ultimum jx191931 pythium nagaii jn630511 pythium okanoganense ab468817 pythium paddicum hq643728 pythium attrantheridium jq898456 pythium intermedium kc602495 pythium intermedium ay083936 pythium sylvaticum ab499723 pythium spinosum ab499701 pythium cylindrosporum hq643516 pythium irregulare jq898464 pythium mamillatum hq643687 pythium spiculum hq643790 pythium debaryanum hq643519 pythium macrosporum ab507408 pythium middletonii hq643694 pythium echinulatum hq643531 pythium radiosum hq643756 pythium mastophorum hq643692 pythium polymastum hq643752 pythium nunn hq643711 pythium perplexum hq643744 pythium undulatum eu240049 pythium dimorphum ay598651 pythium undulatum kc602496 pythium undulatum kc602497 pythium helicandrum ay598653 pythium prolatum hq643754 pythium anandrum hq643435 pythium anandrum kc602489 pythium anandrum kc602490 pythium senticosum hq643773 pythium amasculinum hq643434 pythium hydnosporum hq643564 pythium aphanidermatum jx462954 pythium deliense hq643522 pythium conidiophorum hq643509 pythium dissimile hq643526 pythium scleroteichum hq643771 pythium catenulatum hq643494 pythium torulosum hq643859 pythium aristosporum hq643447 pythium volutum hq643971 pythium graminicola hq643545 pythium inflatum hq643566 pythium monospermum hq643697 pythium pachycaule ef583439 pythium flevoense hq643538 pythium aquatile hq643445 pythium oopapillum hq643717 pythium coloratum hq643506 pythium lutarium jq898467 pythium diclinum jq898459 pythium diclinum kc602491 pythium dissotocum jn863965 pythium marinum ay598689 100/100 100/100 88/99 100/100 100/100 77/98 100/ 100 100/100 98/100 88/98 */75 100/100 99/100 100/100 99/100 100/100 100/10096/100 95/100 95/100 96/100 */83 100/100 100/100 100/100 */96 */98 100/100 97/100 */96 */89 87/100 92/100 99/100 100/100 */100 100/100 0.2 f h b fig. 1 phylogram obtained from the analyses of its sequence data, revealing the identity of pythium spp. isolated from soil in oak stands in the southern part of poland. sequences obtained during this study are presented in bold type. the phylogram was obtained from maximum likelihood (ml) analyses. the bootstrap values (>75%) for ml and posterior probabilities (>75%) that were obtained from bayesian (bi) analyses are presented at nodes as follows: ml/bi. * bootstrap values <75%. 7 of 10© the author(s) 2015 published by polish botanical society acta mycol 50(1):1052 jankowiak et al. / pythium species occurring in oak forests pythium species in oak stands is in accordance with results from previous studies [10,11,13,14]. of all the pythium species recorded in association with the rhizosphere soil of oak stands in europe, p. anandrum and p. undulatum are the most commonly reported. consistent with jung et al. [10], balcì and halmshlager [11] and jönsson et al. [13], we isolated p. anandrum most frequently. however, in contrast to these studies but similar to cordier et al. [14], we additionally relatively often isolated species resembling phytopythium citrinum. the level of pythium diversity found in our survey was certainly underestimated because the typically selective medium for pythiumlutariumjq898467 pythiumdiclinumjq898459 phytophthoraramorumhq643339 phytophthoralateralishq875390 phytopythiummercuriale dq916350 phytopythiumkandeliae kj399961 phytopythiummirpurense kj831614 phytopythiumostracodes hq643395 phytopythiummegacarpumhq643388 phytopythiumboreale hq643372 phytopythiumoedochilumhq643394 phytopythiumsp. 3 kc602494 phytopythiumsp. ab690623 phytopythium sp. 2 kc602493 phytopythiummontanumhq643391 phytopythiumcarbonicumhq643373 pythium sp. eu669081 phytopythiumlitorale jq898465 pythiumsterilum eu240096 phytopythiumcitrinum kc602483 pythiumsp. dq403787 phytopythiumcitrinum kc602484 phytopythiumcitrinumhq643375 phytopythiumcitrinum kc602480 phytopythiumcitrinum kc602482 phytopythiumcitrinum kc602481 phytopythiumcf. citrinum a kc602486 phytopythiumcf. citrinum a kc602485 phytopythium cf. citrinum b kc602487 phytopythium cf. citrinum b kc602488 pythiumsp. ef152506 pythiumsp. ef152505 phytopythiumdelawarense eu339312 phytopythiumhelicoides hq643382 phytopythiumchamaehyphon hq643374 phytopythium sp. 1 kc602492 pythiumsp. jn863966 phytopythiumvexans jx074739 100/100 100/ 100 100/100 */92 75/99 */87 83/91 78/84 82/100 100/100 96/100 */91 100/100 98/100 100/100 */81 */95 100/100 0.5 100/100 100/ 100 100/ 100 fig. 2 phylogram obtained from the analyses of its sequence data, revealing the identity of phytopythium spp. isolated from soil in oak stands in the southern part of poland. sequences obtained during this study are presented in bold type. the phylogram was obtained from maximum likelihood (ml) analyses. the bootstrap values (>75%) for ml and posterior probabilities (>75%) that were obtained from bayesian (bi) analyses are presented at nodes as follows: ml/bi. * bootstrap values <75%. 8 of 10© the author(s) 2015 published by polish botanical society acta mycol 50(1):1052 jankowiak et al. / pythium species occurring in oak forests pythium has not been used. therefore further studies will be needed to fully characterize the oak-associated pythium species. among the species of the genus pythium, only p. undulatum is known to be pathogenic to oak [16]. weber et al. [3] showed also the aggressiveness of p. undulatum on roots of a. procera and p. menziesii. in addition, shafizadeh and kavanagh [25] have also shown the pathogenicity of p. undulatum on picea sitchensis (bong.) carr., picea abies (l.) karst. and pinus contorta dougl. ex loud. in the present study, this root pathogen has been recorded only on one site suggesting that rather does not play important role in the destruction of root systems of oaks. however, the majority of phytopythium spp. and pythium spp. isolates have been obtained from declining stands indicating a possible association between the presence of these organisms and health status of trees. similar relationships have been revealed for soil-borne phytophthora spp. in several european countries [10–12,26,27]. it was shown that phytopythium and pythium species are widespread on a range of different soil textures with a mean soil ph (cacl2) between 3.29 and 5.39. our results resembled those of jung et al. [10] and balcì and halmshlager [11] who mentioned that pythium species in germany and austria occurred in similar site conditions like phytophthora species in oak forests. tab. 3 isolation frequencies of phytopythium spp. and pythium spp. from soil samples in oak stands in poland and soil ph ranges. taxon ph range (cacl2) number of positive soil samples frequency of isolation (%) percent of positive stands site number phytopythium citrinum 3.68–3.99 4 2.3 6.9 26, 27 phytopythium cf. citrinum a 5.39 1 0.6 4.2 10 phytopythium cf. citrinum b 4.82–5.39 3 1.7 6.9 9,10 pythium anandrum 3.75–4.22 7 4.0 13.8 2, 15, 24, 26 pythium diclinum 3.29 1 0.6 4.2 24 pythium intermedium 5.18 1 0.6 4.2 25 pythium undulatum 4.22 1 0.6 4.2 15 phytopythium sp. 1 3.99–5.39 4 2.3 6.9 10, 26 phytopythium sp. 2 4.82 2 1.1 4.2 9 phytopythium sp. 3 4.82–5.39 5 2.9 6.9 9, 10 total 23 13.2 27.6 2, 9, 10, 15, 24, 25, 26, 27 references 1. kirk pm, cannon pf, minter dw, staplers ja. ainsworth & bisby’s dictionary of the fungi. 10th ed. wallingford: cab international; 2008. 2. webster j, weber rws. introduction to fungi. 3rd ed. cambridge: cambridge university press; 2007. http://dx.doi.org/10.1017/cbo9780511809026 3. weber rws, sulzer fl, haarhaus m. pythium undulatum, cause of root rot of abies procera christmas trees and pseudotsuga menziesii in northern germany. mycol prog. 2004;3(3):179–188. http://dx.doi.org/10.1007/s11557-006-0087-7 4. van der plaats-niterink aj. monograph of the genus pythium. baarn: centraalbureau voor schimmelcultures; 1981. 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forest research institute, poland; https://orcid.org/0000-00024363-704x funding the study was carried out as part of the tasks financed from the subsidy of the ministry of science and higher education for the department of algology and mycology, faculty of biology and environmental protection, university of lodz. competing interests no competing interests have been declared. copyright notice © the author(s) 2021. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. short communication in mycology first record of slime molds in biebrza national park (ne poland) dominika małgorzata ślusarczyk * department of algology and mycology, faculty of biology and environmental protection, university of lodz, poland * to whom correspondence should be addressed. email: dominika.slusarczyk@biol.uni.lodz.pl abstract is paper provides the first recorded data of slime molds in biebrza national park (ne poland). in total, 16 species of myxomycetes belonging to nine genera were observed. keywords myxomycetes; ecology; protected area; ne poland 1. introduction myxomycetes are a small group of eukaryotic organisms that includes approximately 1,000 species worldwide. ey are characterized by a complex life cycle that is distinguished by the presence of a plasmodium stage and the formation of sporocarps (baba & sevindik, 2018; lado, 2005–2021; stephenson & rojas, 2017). poland has approximately 250 species of myxomycetes (drozdowicz et al., 2003), the information on which remains fragmented. however, slime molds have been studied in several national parks (drozdowicz, 1997, 2004, 2009, 2014; komorowska & drozdowicz, 1996; magiera & drozdowicz, 2004; narkiewicz et al., 2013; panek & romański, 2010; salamaga et al., 2016). e ecology of northeast (ne) poland is rich and varied, with many species of plants, animals, and fungi. ey are protected in four national parks: białowieża national park, narew national park, biebrza national park (bbnp), and wigry national park. bbnp was established in 1993; it is the largest national park in poland, covering 59.223 ha with a 66.824 ha buffer zone. e park is unique within europe in that it encompasses an entire river valley, from its sources to its mouth. e river valley itself comprises a number of habitats preserved in an almost unchanged state, organized according to natural longitudinal and transversal zones with their corresponding plant communities as well as a large complex of fens. some of the most predominant habitats in biebrza valley are peatlands with swamp forests comprised mainly of alder and birch (dyrcz & werpachowski, 2005). e first data on slime molds in the ne poland region were presented by krzemieniewska (1957, 1960). e samples were collected from the białowieża forest as part of a project for obtaining herbarium materials (i.e., myxomycetes) from different regions in poland. en, drozdowicz (2014) conducted research there, where she assigned 103 species of slime molds. a field study by panek and romański (2010) in wigry national park (ne poland) yielded an interesting and rich collection of slime molds, including seven species new to the country. e present data on bbnp were obtained as part of a mycological study undertaken by members of the polish mycological society. e results of the mycological study were previously presented by kujawa et al. (2012, 2015) and ruszkiewicz-michalska et al. (2012, 2015). acta mycologica / 2021 / volume 56 / article 564 publisher: polish botanical society 1 https://doi.org/10.5586/am.564 https://orcid.org/0000-0002-4363-704x https://orcid.org/0000-0002-4363-704x https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0002-9357-4497 mailto:dominika.slusarczyk@biol.uni.lodz.pl ślusarczyk / first record of slime molds in biebrza national park figure 1 localization on observation sites in the biebrza national park (according to kujawa et al., 2012, modified). 2. material and methods e study was carried out in the central area of bbnp (figure 1) from august 28 to september 1, 2012, and from august 24 to august 29, 2013. e collections of slime molds were primarily acquired from two protected areas: grzędy and kapice. tilio-carpinetum forest associations were identified within these sites. e multispecies stand included carpinus betulus, tilia cordata, acer platanoides, and betula pendula. e field layer was composed of anemone nemorosa, stellaria holostea, galeobdolon luteum, aegopodium podagraria, asarum europaeum, pulmonaria obscura, and lilium martagon (dyrcz & werpachowski, 2005). macromorphological and micromorphological analyses were performed. e specimens were observed with a nikon smz-10 binocular microscope and a nikon eclipse e-200 light microscope. e identification of species was carried out according to specialistic literature, e.g., neubert, nowotny, and baumann (1993), neubert, nowotny, baumann, and marx (1995, 2000), ing (1999), and nannenga-bremekamp (1991). e nomenclature followed those accepted by lado (2005–2021). e specimens documented in the study were preserved at the fungal collection of the herbarium universitatis lodziensis. 3. list of species a total of 16 species belonging to nine genera were identified. is investigation broadly contributes to research in this area by providing the first recorded list of taxa in the bbnp. ese results show that even short-term investigations in areas of unique natural value can contribute to the development of knowledge on the diversity and distribution of slime mold species in poland. arcyria cinerea (bull.) pers., on decaying fallen twigs, kapice district, 2012-08-28; leg. & det. dś acta mycologica / 2021 / volume 56 / article 564 publisher: polish botanical society 2 ślusarczyk / first record of slime molds in biebrza national park a. denudata (l.) wettst., on logs, kapice and grzędy districts, 2012-08-28–2012-08-29, leg. & det. dś ceratiomyxa fruticulosa (o. f. müll.) t. macbr., on a strongly decayed trunk, grzędy district, 2013-08-27, leg. m. ruszkiewicz-michalska, det. dś diachea leucopodia (bull.) rostaf., on fallen small twigs, kapice district, 2012-08-28, leg. & det. dś fuligo leviderma h. neubert, nowotny & k. baumann, on fallen branches of betula sp., kapice district, 2012-08-28, leg. & det. dś f. luteonitens l. g. krieglst. & nowotny, on fallen branches of betula sp., grzędy district, 2013-08-27, leg. j. szkodzik, det. dś hemitrichia serpula (scop.) rostaf. ex lister, on decaying trunks, grzędy district, 2012-08-29, leg. m. wrzosek, det. dś lycogala conicum pers., on decaying wood, kapice district, 2012-08-28, leg. & det. dś l. epidendrum (l.) fr., on coniferous wood, kapice district, 2012-08-28, leg. j. szkodzik, det. dś metatrichia vesparia (batsch) nann.-bremek. ex g. w. martin & alexop., on strongly decayed wood of deciduous tree, kapice district, 2012-08-28, leg. & det. dś mucilago crustacea p. micheli ex f. h. wigg., stem of convallaria majalis, kapice district, 2012-08-28, leg., m. wrzosek, det. dś physarum bivalve pers., on fallen leaves, grzędy district, 2013-08-27, leg. j. szkodzik, det. dś stemonitis fusca roth, on coniferous wood, grzędy district, 2012-08-29, leg. & det. dś s. pallida wingate, on deciduous wood, grzędy district, 2012-08-29, leg. & det. dś trichia favoginea (batsch) pers., on fallen branches, kapice district, 2012-08-28, leg. & det. dś; grzędy district, 2013-08-27, leg. j. szkodzik, det. dś tubifera ferruginosa (batsch) j. f. gmel., on decayed wood, grzędy district, 2013-08-27, leg. m. ruszkiewicz-michalska, det. dś acknowledgments e author would like to thank marta wrzosek, małgorzata ruszkiewicz-michalska, and jarosław szkodzik, who kindly provided their myxomycetes vouchers, as well as two anonymous reviewers for their constructive comments on an earlier version of this manuscript. references baba, h., & sevindik, m. (2018). e roles of myxomycetes in ecosystems. journal of bacteriology & mycology: open access, 6(3), 165–166. https://doi.org/10.15406/jbmoa.2018.06.00197 drozdowicz, a. (1997). studies on myxomycetes in the pieniny national park. i. new species for the pnp. acta mycologica, 32(2), 287–291. https://doi.org/10.5586/am.1997.025 drozdowicz, a. (2004). materiały do chorologii śluzowców w bieszczadzkim parku narodowym [materials for myxomycetes chorology in the bieszczady national park]. roczniki bieszczadzkie, 13, 261–276. drozdowicz, a. (2009). śluzowce [myxomycetes]. in a. górecki, & b. zemanek (eds.), magurski park narodowy. monografia przyrodnicza [magura national park] (pp. 91–96). magurski park narodowy; uniwersytet jagielloński. drozdowicz, a. (2014). myxomycetes of the białowieża forest. białowieski park narodowy. drozdowicz, a., ronikier, a., stojanowska, w., & panek, e. (2003). myxomycetes of poland – a checklist. w. szafer institute of botany, polish academy of sciences. dyrcz, a., & werpachowski, c. (2005). przyroda biebrzańskiego parku narodowego. monografia [nature of the biebrza national park. a monograph]. biebrzański park narodowy. acta mycologica / 2021 / volume 56 / article 564 publisher: polish botanical society 3 https://doi.org/10.15406/jbmoa.2018.06.00197 https://doi.org/10.5586/am.1997.025 ślusarczyk / first record of slime molds in biebrza national park ing, b. (1999). e myxomycetes of britain and ireland. an identification book. e richmond publishing. komorowska, h., & drozdowicz, a. (1996). śluzowce [slime molds]. in z. mirek (ed.), przyroda tatrzańskiego parku narodowego [e nature of the tatra national park] (pp. 405–412). tatrzański park narodowy. krzemieniewska, h. (1957). spis śluzowców zebranych w latach 1955–1956 [list of slime molds collected in 1955–1956]. acta societatis botanicorum poloniae, 26(4), 785–811. https://doi.org/10.5586/asbp.1957.041 krzemieniewska, h. (1960). śluzowce polski na tle flory śluzowców europejskich [polish slime molds against the background of european slime mold flora]. pwn. kujawa, a., gierczyk, b., domian, g., wrzosek, m., stasińska, m., szkodzik, j., leski, t., karliński, l., pietras, m., dynowska, m., henel, a., ślusarczyk, d., & kubiak, d. (2015). preliminary studies of fungi in the biebrza national park. part iv. macromycetes – new data and the synthesis. acta mycologica, 50(2), article 1070. https://doi.org/10.5586/am.1070 kujawa, a., wrzosek, m., domian, g., kędra, k., szkodzik, j., rudawska, m., leski, t., karliński, l., pietras, m., gierczyk, b., dynowska, m., ślusarczyk, d., kałucka, i., & ławrynowicz, m. (2012). preliminary studies of fungi in the biebrza national park. ii. macromycetes. acta mycologica, 47(2), 235–264. https://doi.org/10.5586/am.2012.026 lado, c. (2005–2021). an on-line nomenclatural information system of eumycetozoa. retrieved september 25, 2020, from https://eumycetozoa.com/ magiera, a., & drozdowicz, a. (2004). śluzowce (myxomycetes) babiogórskiego parku narodowego [slime molds of the babia góra national park]. in b. w. wołoszyn, a. jaworski, & j. szwagrzyk (eds.), babiogórski park narodowy. monografia przyrodnicza [babia góra national park. nature monograph] (pp. 315–332). w. szafer institute of botany, polish academy of sciences. nannenga-bremekamp, n. e. (1991). a guide to temperate myxomycetes. biopress. narkiewicz, c., pusz, w., kita, w., & panek, e. (2013). grzyby i śluzowce [fungi and slime molds]. in p. knapik, r. migoń, & a. raj (eds.), przyroda karkonoskiego parku narodowego [nature of the karkonosze national park] (pp. 339–358). karkonoski park narodowy. neubert, h., nowotny, w., & baumann, k. (1993). die myxomyceten deutschlands und des angrenzenden alpenraumes unter besonderer berücksichtigung österreichs [myxomycetes in germany and the alpine region with special focus on austria] (vol. 1). karlheinz baumann verlag. neubert, h., nowotny, w., baumann, k., & marx, h. (1995). die myxomyceten deutschlands und des angrenzenden alpenraumes unter besonderer berücksichtigung österreichs [myxomycetes in germany and the alpine region with special focus on austria] (vol. 2). karlheinz baumann verlag. neubert, h., nowotny, w., baumann, k., & marx, h. (2000). die myxomyceten deutschlands und des angrenzenden alpenraumes unter besonderer berücksichtigung österreichs [myxomycetes in germany and the alpine region with special focus on austria] (vol. 3). karlheinz baumann verlag. panek, e., & romański, m. (2010). śluzowce myxomycetes [slime molds]. in l. krzysztofiak (ed.), śluzowce myxomycetes, grzyby fungi i mszaki bryophyta wigierskiego parku narodowego [slime molds, fungi and bryophytes of the wigry national park] (pp. 9–84). stowarzyszenie “człowiek i przyroda”. ruszkiewicz-michalska, m., bałazy, s., chełkowski, j., dynowska, m., pawłowska, j., & sucharzewska, e. (2015). preliminary studies of fungi in the biebrza national park (ne poland). part iii. micromycetes – new data. acta mycologica, 50(2), article 1067. https://doi.org/10.5586/am.1067 ruszkiewicz-michalska, m., tkaczuk, c., dynowska, m., sucharzewska, e., szkodzik, j., & wrzosek, m. (2012). preliminary studies of fungi in the biebrza national park (ne poland). i. micromycetes. acta mycologica, 47(2), 213–234. https://doi.org/10.5586/am.2012.026 salamaga, a., grzesiak, b., wolski, g. j., kochanowska, m., & kochanowski, j. (2016). preliminary investigations into the slime moulds (myxogastria) in the “bory tucholskie” national park. acta mycologica, 51(1), article 1077. https://doi.org/10.5586/am.1077 stephenson, s. l., & rojas, c. (2017). introduction. in s. l. stephenson, & c. rojas (eds.), myxomycetes: biology, systematics, biogeography, and ecology (pp. 17–20). academic press. https://doi.org/10.1016/b978-0-12-805089-7.00017-2 acta mycologica / 2021 / volume 56 / article 564 publisher: polish botanical society 4 https://doi.org/10.5586/asbp.1957.041 https://doi.org/10.5586/am.1070 https://doi.org/10.5586/am.2012.026 https://eumycetozoa.com/ https://doi.org/10.5586/am.1067 https://doi.org/10.5586/am.2012.026 https://doi.org/10.5586/am.1077 https://doi.org/10.1016/b978-0-12-805089-7.00017-2 first record of slime molds in biebrza national park (ne poland) 1 introduction figure 1 2 material and methods 3 list of species acknowledgments references 2014-11-20t23:15:48+0000 polish botanical society 2014-11-20t23:20:52+0000 polish botanical society 2014-11-20t23:18:48+0000 polish botanical society 2014-11-20t23:17:03+0000 polish botanical society 2014-11-20t23:19:47+0000 polish botanical society 2014-11-20t23:21:24+0000 polish botanical society additions to the lichen biota of the sudety mountains. i. records from the karkonosze mountains 1 of 9published by polish botanical society acta mycologica original research paper additions to the lichen biota of the sudety mountains. i. records from the karkonosze mountains maria kossowska1*, wiesław fałtynowicz1, monika dimos-zych1, hanna fałtynowicz2, katarzyna patejuk3, amelia piegdoń4, magdalena buksakowska1, paweł jarema1 1 department of botany, institute of environmental biology, university of wrocław, kanonia 6/8, 50-328 wrocław, poland 2 division of fuels chemistry and technology, faculty of chemistry, wrocław university of technology, gdańska 7/9, 50-344 wrocław, poland 3 faculty of life sciences and technology, wrocław university of environmental and life sciences, pl. grunwaldzki 24a, 50-363 wrocław, poland 4 jan grodek state vocational academy in sanok, mickiewicza 21, 38-500 sanok, poland * corresponding author. email: maria.kossowska@uwr.edu.pl abstract records of 10 rare and noteworthy lichen species in poland have been presented. four species, japewia subaurea, myriolecis persimilis, palicella filamentosa, and scoliciosporum sarothamni are new to the polish part of the sudetes. anisomeridium polypori and pyrenula coryli are new species to the karkonosze mountains. keywords lichenized fungi; lichen diversity; epiphytic lichens; lower silesia introduction the sudety mountains are one of regions with the richest and the best-known lichen biota in poland, with more than 1,000 species recorded to date (kossowska, unpublished). nevertheless, detailed studies still provide new data about the occurrence and distribution of lichen species (e.g., [1–3]). in addition, there are still some sudetian mountain ranges where this group of organisms is poorly recognized. the aim of this new series of publications is to expand our knowledge and to provide the most up-todate information about interesting and rare lichen taxa in poland. in this paper, 10 noteworthy lichen species have been presented. all these species were found in the highest sudetian range, the karkonosze mountains. four of them, japewia subaurea, myriolecis persimilis, palicella filamentosa, and scolicosporum sarothamni have been reported for the first time from the polish part of the sudetes. two others, anisomeridium polypori and pyrenula coryli, are new to the karkonosze mountains. material and methods the species were recorded mainly during the third edition of the monitoring of epiphytic lichens on permanent sample plots in the karkonoski national park, carried out in 2016. the methodology of the monitoring used has been described in detail in [4]. collected specimens were identified using standard techniques, with an aid of a nikon smz-u stereoscope and a nikon eclipse e600 light microscope. anatomical doi: 10.5586/am.1113 publication history received: 2018-05-30 accepted: 2018-10-17 published: 2018-12-17 handling editor piotr zaniewski, faculty of forestry, warsaw university of life sciences – sggw, poland authors’ contributions mk: identification of species, writing the manuscript; wf: identification of species; mdz, hf: field works, tlc analyses; kp, ap, mb, pj: field works funding the study was supported by polish state forests (fundusz leśny), project “examination of epiphytic lichens on permanent circular sample plots in forest ecosystems of the karkonoski national park – the third edition of monitoring”. competing interests no competing interests have been declared. copyright notice © the author(s) 2018. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation kossowska m, fałtynowicz w, dimos-zych m, fałtynowicz h, patejuk k, piegdoń a, et al. additions to the lichen biota of the sudety mountains. i. records from the karkonosze mountains. acta mycol. 2018;53(2):1113. https://doi. org/10.5586/am.1113 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:maria.kossowska%40uwr.edu.pl?subject=additions%20to%20the%20lichen%20biota%20of%20the%20sudety%20mountains.%20i.%20records%20from%20the%20karkonosze%20mountains https://doi.org/10.5586/am.1113 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1113 https://doi.org/10.5586/am.1113 2 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(2):1113 kossowska et al. / additions to the sudetian lichen biota i characters were measured from hand-cut sections. brief descriptions of the species are based on own observations and measurements. chemical properties of the thalli were examined using standard reagents: 10% potassium dioxide (k), sodium hypochlorite (c), p-phenylenediamine in ethanol (pd), and lugol’s iodine (i). a content of secondary metabolites was determined by thin layer chromatography (tlc) in solvent a [5]. localities of each species are arranged according to atpol grid square system [6]. collected material has been deposited in the lichen herbarium of the first author. names of the taxa are given according to [7]. results anisomeridium polypori (m. b. ellis & everh.) m. e. barr. thallus very thin and inconspicuous, greenish. ascomata perithecia, not seen in collected material. specimen identified on the basis of characteristic, conical pycnidia containing macroconidia. pycnidia numerous, black, 0.1–0.15 mm diam., sessile, with gelatinous, white tendril containing elipsoid conidia ejected from the ostiole. anisomeridium polypori is one of the species with rapidly increasing number of records in poland. in the twentieth century, it was reported only from a northern part of the country [8]; now this species has been identified at many sites, including central and southern poland [9–13]. it is difficult to confirm if this species is actually spreading or the increasing number of localities is the result of a more thorough examination of the lichen biota. the reported site of anisomeridium polypori is the first one in the polish part of the karkonosze mountains. specimen examined. eb 80 – eastern karkonosze mts, ne slope of the kopa mt, between łomniczka and wilczy potok streams, 50°45'29.3" n / 15°44'53.7" e, on the bark of betula pendula, july 6, 2016, leg. a. piegdoń & h. fałtynowicz. cladonia caespiticia (pers.) flörke thallus composed of irregularly incised, erect squamules forming cushions. podetia very short, without algal layer, translucent when wet. apothecia pale brown to brown, either on podetia or sessile directly on squamules. brown pycnidia also present on the upper surface and the edges of squamules. thallus turns red with pd because of the presence of fumaroprotocetraric acid. although this species is widespread in poland, it is rather rare and considered endangered (category en in polish red list of lichens [14]). contemporarily, it was recorded in several mountain ranges belonging to the sudetes, namely in the masyw śnieżnika (śnieżnik massif ) and bialskie mountains [15], pogórze kaczawskie (kaczawskie foothills) [16], kotlina jeleniogórska (jelenia góra basin) [17], and stołowe mountains [13]. the new findings presented here are the first ones in the karkonosze mountains since the beginning of the twentieth century [18,19]. specimens examined. ea 79 – pogórze karkonoskie (karkonosze foothills), dolina choińca (choiniec valley), 50°49'51.7" n / 15°39' 11.1" e, on the base of picea abies, july 31, 2016, leg. a. piedgoń & h. fałtynowicz; e slope of żar mt, 50°49'45.1" n / 15°38'55.9" e, on the base of fagus sylvatica, july 31, 2016, leg. a. piedgoń & h. fałtynowicz. japewia subaurifera muhr & tønsberg thallus almost entirely sorediate, yellowish brown. soralia initially discrete, but soon coalescing to form thick, leprose crust. soredia granular, externally brown, internally yellow (visible in abraded soralia), with characteristic oil droplets. brown pigment of soredia became k+ fuscous. apothecia are not present in the collected specimen. 3 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(2):1113 kossowska et al. / additions to the sudetian lichen biota i japewia subaurifera was reported in poland for the first time from an upper montane belt in the tatra mountains [20]. until now, it was known in the country to be present only in montane spruce forests in the carpathians [21]; however, in the czech republic, this species was recorded also in the sudety mountains [22]. although in scandinavian boreal forests this species grows on various deciduous trees, like betula spp., sorbus aucuparia and alnus incana [23], in poland and the czech republic it was collected only from conifers. the new finding extends the range of japewia subaurifera on the polish part of the sudetes; it is also the first polish record of this species on a deciduous phorophyte. specimen examined. ea 79 – western karkonosze mts, slope above jagniątków, 50°48'32.6" n / 15°36'36.9" e, on the bark of betula pendula, july 22, 2016, leg. k. patejuk & p. jarema. mycoblastus sanguinarius (l.) norman thallus crustose, very thick, with irregular, verrucose surface, pale grey. apothecia numerous, big (up to 1 mm diam.), black and shining, at first plane but soon becoming convex. easily distinguished by characteristic carmine-red pitts visible when thallus or apothecia are damaged. the same red pigment is also present in the hypothecium. hymenium aeruginose throughout in examined specimen. spores large, 90–95 µm, with thick walls. thallus reacts k+ and pd+ faintly yellow. mycoblastus sanguinarius is a rather rare boreal lichen, occurring in the northern and southern, mountainous regions of poland [8]. contemporarily, in the sudetes, it was found only in the karkonosze mountains [24] and the masyw śnieżnika [25,26]. specimen examined. ea 78 – western karkonosze mts, upper part of the dolina kamieńczyka (kamieńczyk valley), 50°47'50.1" n / 15°30'02.4" e, on the bark of picea abies, july 28, 2016, leg. m. dimos-zych & m. buksakowska. myriolecis persimilis (th. fr.) śliwa, zhao xin & lumbsch thallus almost inapparent, membranaceous, grey. apothecia partly single, partly clustered in groups of two–three, widely attached to the substratum. thalline margin olive-brown, slightly pruinose at the edge. discs pale brown to chocolate, glossy. hymenium ca. 50 µm, with olive brown epithecium. spores elipsoid, 10–12 × 5–6 µm. myriolecis persimilis is very often reported in recent lichenological literature and seems to be common in poland. probably it has been overlooked earlier because of growing predominantly on twigs and small branches [27]. however, all polish records to date were from northern and central part of the country [28–34]. the new finding is the first from the mountainous areas in the south of poland and the species is new in the polish sudetes. specimen examined. ea 78 – western karkonosze mts, slope gawry below the mały śnieżny kocioł (mały śnieżny cirque), 50°47'13.2" n / 15°33'37.7" e, on the bark of sorbus aucuparia, july 30, 2016, leg. m. dimos-zych & m. buksakowska. palicella filamentosa (stirt.) rodr. flakus & printzen thallus continuous or cracked-areolate, white to pale greenish-grey. apothecia numerous, usually crowded, variable in color, from pale brown to almost black. thalline exciple present in early stages, but soon excluded and only true exciple visible. hymenium ca. 50 µm, with brown epithecium. hypothecium and exciple colorless. spores single, ellipsoid, 12–14 × 4–5 µm. thallus reacts k+ and pd+ yellow because of the presence of atranorin. this is a member of newly described genus palicella [35], in former polish literature known mainly as lecidea ramulicola (h. magn.) hillman or lecanora ramulicola (h. 4 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(2):1113 kossowska et al. / additions to the sudetian lichen biota i magn.) printzen & p. f. may. the species has been considered rare in poland. however, recent detailed studies [36] showed that it was quite frequent in the country and only had been misidentified or generally misunderstood. to date, the known distribution of palicella filamentosa in poland included areas from the baltic coast to the western carpathians; the species was recorded also in the czech part of the sudety mountains near the polish border [37]. here, it is reported as new to the polish sudetes. the specimens of p. filamentosa were collected on 19 dispersed sites in the karkonosze mountains; the species seems to be widespread and frequent on spruce in upper montane belt of this mountain range. probably it had been collected earlier but not properly identified because of nomenclatural and taxonomical confusion concerning this taxon in the polish literature [36]. selected specimens examined. ea 78 – western karkonosze mts, “zielony klin” – n slope of mumlawski wierch mt, 50°48'02.1" n / 15°28'12.9" e, on the bark of picea abies, july 30, 2016, leg. a. piegdoń & h. fałtynowicz; n slope of czeskie kamienie mt, 50°46'54.9" n / 15°35'25.4" e, on the bark of picea abies, july 19, 2016, leg. a. piegdoń & h. fałtynowicz.; ea 89 – western karkonosze mts, n slope of ptasi kamień mt, 50°46'04.5" n / 15°37'44.4" e, on the bark of picea abies, july 20, 2016 leg. m. dimoszych, a. piegdoń & h. fałtynowicz; eastern karkonosze mts, valley of pląsawa stream above polana, 50°45'46.8" n / 15°42'04.8" e, on the bark of picea abies, july 15, 2016, leg. k. patejuk & h. fałtynowicz; eb 80 – eastern karkonosze mts, sowia przełęcz (sowia pass), 50°44'51.3" n / 15°47'12.3" e, on the bark of picea abies, july 8, 2016, leg. a. piegdoń & h. fałtynowicz; kowarski grzbiet (kowary ridge), 50°45'17.9" n / 15°48'27.8" e, on the bark of picea abies, july 7, 2016, leg. k. patejuk & p. jarema. porina leptalea (durieu & mont.) a. l. sm. thallus very thin, smooth, dark olive green. ascomata perithecia, one half immersed in the substrate, reddish brown and glossy, 0.2–0.3 mm in diam. involucrellum orange in section, containing algal cells (trentepohlia sp.). excipulum colorless, ca. 200 µm wide. spores one–three-septate, fusiform, 20 × 4–5 µm. easily distinguished from the other corticolous species of the genus, p. aenea, by the color of perithecia. porina leptalea is considered rare and endangered in poland (category en on the polish red list of lichens [11]). all the known localities of this species are concentrated in mountainous parts of the country [38–42]. in the sudety mountains, it is known from several contemporary localities in masyw śnieżnika [43], pogórze kaczawskie [16], and karkonosze mountains [44]. new record suggests that this species is probably widely distributed, at least in a southern part of poland, but may have been overlooked. specimen examined. ea 89 – eastern karkonosze mts, kocioł łomniczki (łomniczka cirque), 50°44'30.6" n / 15°44'08.8" e, on the base of picea abies, intermixed with micarea botryoides, july 11, 2016, leg. m. dimos-zych & m. buksakowska. pyrenula coryli a. massal. “thallus” inconspicuous, greyish, apparently not lichenized. perithecia numerous, ca. 0.2 mm diam, with colorless excipulum and laterally extended involucrellum. spores 15–17 × 5 µm, three-septate, with strongly thickened walls and lenticular cells. no pycnidia visible in the collected material. this species is often considered nonlichenized and named mycopyrenula coryli (a. massal.) vain. however, scattered cells of the photobiont are sometimes present in the “thallus” [45,46]. pyrenula coryli is rarely reported in the lichenological literature; in poland, only historical data from the nineteenth and early half of twentieth centuries are available [8]. the new record is the first one from the karkonosze mountains. specimen examined. ea 78 – western karkonosze mts, slope gawry below mały śnieżny kocioł, 50°47'13.2" n / 15°33'37.7" e, on the bark of sorbus aucuparia, july 30, 2016, leg. m. dimos-zych & m. buksakowska. 5 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(2):1113 kossowska et al. / additions to the sudetian lichen biota i pyrenula nitida (weigel) ach. thallus continuous or cracked, partly immersed in the substrate, olive, with dot-like pseudocyphellae. perithecia big, up to 0.8 mm diam., black and shiny. outer part of hymenium with orange-brown mass of anthraquinones, k+ purple-red. spores threeseptate, with strongly thickened walls and lenticular cells, pale brown, 20–24 × 6–7 µm. pycnidia numerous, with simple, curved conidia. thallus k+ orange. pyrenula nitida is widespread in poland [8] but has not been found frequently. cieśliński et al. [14] considered it endangered in the country (cat. vu). in the sudety mountains, this species used to be common [18], but in the twenty-first century, it was reported only from several sites in the masyw śnieżnika [15] and stołowe mountains [13]. the localities reported here are the first from the polish part of the karkonosze mountains. specimens examined. ea 79 western karkonosze mts, dolina czerwienia (czerwień valley), on bark of fagus sylvatica, july 2011, leg. k. kobylnik; eb 80 – eastern karkonosze mts, valley of łomniczka stream above karpacz-wilcza poręba, 50°45'42.7" n / 15°45'39.3" e, on the bark of betula pendula, july 7, 2016, leg. m. dimos-zych, m. buksakowska, k. patejuk, p. jarema, a. piegdoń & h. fałtynowicz. scoliciosporum sarothamni (vain.) vězda thallus thin and warted, olive brown, sorediate. soralia initially discrete, punctiform, 0.1 mm wide, later coalescing and becoming contiguous. soredia minutely granular, yellowish green, c+ faintly red. apothecia numerous in examined material, pale brown to almost black, 0.2–0.3 mm diam. hymenium 50–60 µm, brown in the upper part. hypothecium colorless. spores curved to s-shaped, spirally arranged in the ascus, mostly three-septate, 20–35 × 2–3 µm. distinguished from the common s. chlorococcum by the presence of soralia reacting with c, and twisted spores [47]. this species was reported from poland for the first time by kowalewska and kukwa [48], who recorded it on several sites in the northern part of the country. since then s. sarothamni has been found in many regions of poland [32,49–52]. it seems to be common but was probably overlooked because of its inapparent habitus. here, it is reported as new to the polish part of the sudetes. specimen examined. eb 80 – eastern karkonosze mts, kowarski grzbiet, n slope of the czoło mt, 50°45'37.5" n / 15°48'58.6" e, on twigs of betula pendula, together with lecanora conizaeoides, july 7, 2016, leg. a. piegdoń & h. fałtynowicz. discussion presented results of the lichenological investigations raised the number of lichen species known from the polish part of the karkonosze mts to 631 [53,54]. the lichen biota of this mountain range is one of the richest and the best known in the mountainous part of poland; in number of recorded species, it yields only to the tatras [55–57]. however, in the latter half of the twentieth century, the karkonosze mts were affected by an ecological disaster. most of the epiphytic lichens disappeared due to the catastrophic level of air pollution. only recently, a slow regeneration of the lichen biota has been observed. this process has been manifested by the increasing number of localities of species sensitive to air pollution, as well as appearance of new taxa and reappearance of regionally extinct ones [58]. presented results seem to confirm this optimistic tendency. almost all new and interesting lichens reported here produce inconspicuous, crustose thalli, easy to overlook during the routine lichenological research. therefore, detailed field investigations, conducted throughout the karkonosze mts as a part of the monitoring program, provided a better opportunity to record them. it is worth emphasizing that almost all reported taxa were recorded on single localities. the only widely distributed new species was palicella filamentosa. 6 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(2):1113 kossowska et al. / additions to the sudetian lichen biota i four of six species reported here as new to the karkonosze mountains, anisomeridium polypori, japewia subaurifera, pyrenula nitida, and scoliciosporum sarothamni, were collected on birches, i.e., trees that are only admixed in mountain forests dominated by spruce. birch is a very interesting phorophyte, offering various microhabitats suitable for different lichen species (young bark is thin and smooth, then starts to peel, finally it is thick and cracked, with various chemical properties [59]). in the karkonosze mts, birch is one of the phorophytes with the most rapidly increasing number of recorded lichen species [60]. in the future, special attention should be paid to trees of this species as potential sources of lichen propagules and centers of recolonization. although relatively well recognized, the lichen biota of the karkonosze mountains, as well as the sudetes as a whole, still requires careful and thorough studies. further detailed researches, especially in the areas and habitats investigated in a lesser extent, may result in more discoveries. because of the general improvement of aerosanitary conditions and the great return of lichens to their former localities, which is observed in various parts of europe [61–63], in the future we can also expect to find species of large, foliose and fruticose macrolichens, known now only from the historical records [60]. acknowledgments we would like to express our gratitude to prof. paweł czarnota (rzeszów, poland) for the identification of palicella filamentosa and helpful information about this species. references 1. kossowska m. new, rare and noteworthy lichens in the giant mountains. biologia. 2011;66(5):755–761. https://doi.org/10.2478/s11756-011-0084-4 2. ossowska e, szczepańska k, kossowska m. new records of parmelia ernstiae and p. serrana (ascomycota, parmeliaceae) in poland. acta mycol. 2015;50(2):1065. https://doi.org/10.5586/am.1065 3. szczepańska k. new records of rare lichenicolous and lichenforming fungi from volcanic rocks in sw poland. acta mycol. 2015;50(1):1056. https://doi.org/10.5586/am.1056 4. kossowska m, szczepańska k, fałtynowicz w, jando k, kowalewska a, dimos m. różnorodność gatunkowa porostów epifitycznych na stałych powierzchniach monitoringowych w karkonoskim parku narodowym. parki narodowe i rezerwaty przyrody. 2007;26(1):3–16. 5. orange a, james pw, white fj. microchemical methods for the identification of lichens. london: british lichen society; 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of the epiphytic lichen flora following air quality improvement in south-west sweden. water air soil pollut. 2004;154:203–211. https://doi.org/10.5586/am.2003.015 https://doi.org/10.5586/am.2012.025 https://doi.org/10.12775/v10090-010-0003-2 9 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(2):1113 kossowska et al. / additions to the sudetian lichen biota i https://doi.org/10.1023/b:wate.0000022967.35036.ca 63. lisowska m. lichen recolonisation in an urban-industrial area of southern poland as a result of air quality improvement. environ monit assess. 2011;179:177–190. https://doi.org/10.1007/s10661-010-1727-6 https://doi.org/10.1023/b:wate.0000022967.35036.ca https://doi.org/10.1007/s10661-010-1727-6 abstract introduction material and methods results anisomeridium polypori (m. b. ellis & everh.) m. e. barr. cladonia caespiticia (pers.) flörke japewia subaurifera muhr & tønsberg mycoblastus sanguinarius (l.) norman myriolecis persimilis (th. fr.) śliwa, zhao xin & lumbsch palicella filamentosa (stirt.) rodr. flakus & printzen porina leptalea (durieu & mont.) a. l. sm. pyrenula coryli a. massal. pyrenula nitida (weigel) ach. scoliciosporum sarothamni (vain.) vězda discussion acknowledgments references 2018-12-17t22:54:09+0000 piotr otręba 2014-11-20t23:18:15+0000 polish botanical society initiation and development of erysiphe necator chasmothecia and their role in the epidemiology of grapevine powdery mildew in southern syria 1 of 11published by polish botanical society acta mycologica original research paper initiation and development of erysiphe necator chasmothecia and their role in the epidemiology of grapevine powdery mildew in southern syria nujoud alimad1, walid naffaa2*, fawaz azmeh3 1 faculty of agriculture, damascus university, sweida branch, sweida, syria 2 department of plant protection, faculty of agriculture, damascus university, 30621 damascus, syria 3 national commission for biotechnology, 30621 damascus, syria * corresponding author. email: walid1851966@yahoo.com abstract powdery mildew caused by erysiphe necator is the most important fungal disease of grapevine in southern syria. the purpose of this study was to determine the development of chasmothecia and their role as a primary inoculum in spring. leaves and/or branches were examined by a stereo binocular from july to december 2014 and 2015. the number of chasmothecia was estimated on both surfaces of the leaves, and their viability was estimated by microscopic examination. during 2 years of survey chasmothecia were detected in 45.5% of vineyards. the initial development of chasmothecia on infected leaves was observed in the second half of july. their numbers increased from july to october, and the sudden reduction at the beginning of november was noted. chasmothecia were formed on 38.7% of infected leaves, with 12.5%, 18.4%, and 7.5% on the upper, under and on both surfaces of infected leaves respectively. chasmothecia were more frequent on the leaf under side (0.6 / leaf ) than on the leaf upper side (0.4 / leaf ), but their occurrence on both sides together was relatively low (0.2 / leaf ), and their numbers were highly variable between vineyards and years. microscopic examination showed that chasmothecia contained 1–5 (usually three) asci with 1–4 (usually three) ascospores in each asci, and 65.6% of chasmothecia were empty. their viability decreased between december and february, with an average viability of 1.2% and 0.2% in march and april, respectively. chasmothecia were not detected on bark and ascospores were not trapped at the beginning of the season. these results indicate that the ascospores have no or little role in the initiation of spring infection. to the best of our knowledge, this is the first report of e. necator chasmothecia development and their role in the initiating infection on grapevine in syria. keywords grapevine; powdery mildew; chasmothecia; primary inoculum; erysiphe necator; syria introduction powdery mildew, caused by the fungus erysiphe necator schwein., is one of the most economically important grapevine diseases, and it causes heavy yield losses as well as a reduction in the quality of the produced wine [1]. in syria, because of the presence of favorable environmental conditions for the development of grapevine powdery mildew, an intense application of fungicides from bud break to the end of season is necessary to control this disease (naffaa, unpublished data). doi: 10.5586/am.1088 publication history received: 2016-08-10 accepted: 2016-12-22 published: 2016-12-28 handling editor tomasz leski, institute of dendrology, polish academy of sciences, poland authors’ contributions na and wn: field survey, samples collection; na: binocular and microscopic examination; na and fa: ascospores and conidia trap funding the authors are grateful to damascus university and its sweida branch for funding this research. competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation alimad n, naffaa w, azmeh f. initiation and development of erysiphe necator chasmothecia and their role in the epidemiology of grapevine powdery mildew in southern syria. acta mycol. 2016;51(2):1088. http://dx.doi. org/10.5586/am.1088 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:walid1851966%40yahoo.com?subject=initiation%20and%20development%20of%20erysiphe%20necator%20chasmothecia%20and%20their%20role%20in%20the%20epidemiology%20of%20grapevine%20powdery%20mildew%20in%20southern%20syria http://dx.doi.org/10.5586/am.1088 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1088 http://dx.doi.org/10.5586/am.1088 2 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator erysiphe necator produces sexual fruiting bodies (chasmothecia) only on the surface of heavily infected tissues of grapevine [2,3]. chasmothecia are produced mainly on leaves, but also on shoots, branches, and berries [4]. the formation of chasmothecia begins within 48 hours when uninucleate hyphae of two compatible mating types come in contact [5,6]. in the early stages of development, chasmothecia are white translucent, turn to light yellow or cream, and finally to dark brown or black at maturation [7]. during this time, the ascocarp also increases in size. the principal sources of primary inoculum of powdery mildew on grapevine are mycelium within dormant buds, and/or ascospores borne in chasmothecia [8]. during winter, e. necator survives as mycelium in dormant buds of grapevine until the following season, where shoots emerged from infected buds are covered with white to grayish mycelium with abundant sporulation, and appear stunted with wrinkled and deformed leaves [9]. these infected shoots are known as “flag shoots”, and carry abundant conidia that cause secondary infections [10]. in most viticultural areas, chasmothecia are the main source of primary inoculum of powdery mildew [4], while they serve as an additional source of primary inoculum when the flag shoots are common [11]. in syria, the first study conducted about the biology of e. necator showed that the flag shoots were the main source of primary inoculum (unpublished data), but the role of chasmothecia in initiating the infection remained unclear. so, the objective of this research was to study the formation, development and occurrence of e. necator chasmothecia, and to highlight their potential contribution to the onset of the disease in southern syria. material and methods chasmothecia on leaves and branches during the period from june until the leaf fall in 2014 and 2015, leaves, branches, and berries showing powdery mildew symptoms were collected and placed in plastic bags from 11 vineyards located in five different sites in sweida (southern syria), where several local grape varieties are grown (tab. 1). in spring, just before bud break, 50 samples of branches were collected, brought to the laboratory, and examined for the chasmothecia formation on the bark [10]. leaves and branches were examined in a stereo binocular for the chasmothecia observation, and the number of chasmothecia was estimated on the upper and under surfaces of the leaves. samples of 30 g of bark were placed in 2-l erlenmeyer flasks containing 1.5 l of water. the flasks were shaken vigorously by hand for 3 min and the suspension was filtered through 120and 150-mesh cobb sieves. the content of the 150-mesh sieve was recovered in 25 ml of water, and the suspension was examined under a light microscope [12]. in fall, senescent leaves still on vines or fallen to the ground were collected, placed in 50 × 50 × 50 cm iron cages covered with polyethylene, and kept in the vineyard during winter to study the development and maturation of chasmothecia and their viability. to evaluate viability, 50 chasmothecia collected from infected leaves of each of five surveyed vineyards, where chasmothecia were observed, were examined in light microscope. chasmothecia were considered viable when they contained at least one viable ascospore. viability of ascospores was evaluated using fluorescein diacetate staining [13]. trapping the released fungal spores in this experiment, for the determination of primary and secondary infection sources, in each vineyard, a 125 × 10 × 2.5 cm wooden stand was excavated at 25, 50, 75, and 100 cm from the soil surface by length of 75 mm (glass slide length) and depth of 2.5 mm. at the beginning of march, three stands were placed between the rows in 3 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator each of four selected locations (kanawat 1st, daher aljabel/1, daher aljabel/2, and daher aljabel/albassah). then, four microscopic slides covered with vaseline were horizontally fixed on every level of each stand for spore trapping. slides were carried to the laboratory every 7 days. then, 192 slides were stained with cotton blue – lactophenol solution, and the number of trapped spores were determined and recorded every week using a light microscope. statistical analysis one-way analysis of variance and tukey’s test were done using spss15 statistical software at (p ≤ 0.05) to compare differences in chasmothecia formation during the period of observation in the same year and between years, in the same vineyard and between vineyards, on the upper, under and both sides of the leaves. results leaves and/or branches were examined for chasmothecia formation from july to december in 2014 and 2015. chasmothecia were detected in 45.5% of vineyards in the 2 years of survey. observation of 110 branches of ‘balady’ and ‘black’ cultivars, severely infected by powdery mildew in 2015, showed the presence of e. necator mycelium and conidia on the surface of infected branches, but chasmothecia were not detected on 1and 2-year old branches. at the time of bud break, in march–april 2014 and 2015, chasmothecia were not found on vine bark, nor ascospores were trapped by the slide traps at the beginning of the season. development of e. necator chasmothecia almost identical results were obtained during the two years of the survey. therefore, only data collected in 2015 are presented here. the initial development of chasmothecia on powdery mildew infected leaves was observed on 20 july 2015 in four vineyards tab. 1 grapevine varieties and vineyard’s location area where the study was conducted. vineyard no. location cultivar training system observed area (m2) altitude 1 kanawat 1st balady earth-trellised “jui” system 4000 1270 2 kanawat 1st black earth-trellised “jui” system 2000 1270 3 kanawat 2nd balady arbour vineyard 2000 1250 4 kanawat 2nd black arbour vineyard 1000 1250 5 kanawat 2nd helwani arbour vineyard 1000 1250 6 kanawat 2nd salty arbour vineyard 400 1250 7 daher aljabel (research center)/1 black lateral cordon 5000 1500 8 daher aljabel (research center)/2 balady head-trained vine 5000 1500 9 daher aljabel (research center)/3 black earth-trellised “jui” system 100 1500 10 daher aljabel (albassa) salty earth-trellised “jui” system 300 1450 11 salkhad balady arbour vineyard 200 1350 4 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator ta b. 2 n um be r of g ra pe vi ne le av es o n w hi ch c ha sm ot he ci a w er e fo rm ed fr om ju ly to d ec em be r 20 15 . d at e n o. o f e xa m in ed le av es n o. o f l ea ve s w it h ch as m oth ec ia o nl y on up pe r su rf ac e % o f l ea ve s w it h ch as m ot he ci a on ly o n up pe r su rf ac e n o. o f l ea ve s w it h ch as m oth ec ia o nl y on un de r su rf ac e % o f l ea ve s w it h ch as m ot he ci a on ly o n un de r su rf ac e n o. o f l ea ve s w it h ch as m oth ec ia o n bo th su rf ac es % o f l ea ve s w it h ch as m ot he ci a on b ot h su rf ac es to ta l n um be r of le av es w it h ch as m ot he ci a % o f l ea ve s w it h ch as m ot he ci a 8/ 7/ 20 15 20 0 0. 0 0 0. 0 0 0. 0 0 0. 0 20 /7 /2 01 5 10 1a 10 .0 1a 10 .0 2b 20 .0 4 40 .0 30 /7 /2 01 5 50 10 a 20 .0 15 b 30 .0 5c 10 .0 30 60 .0 6/ 8/ 20 15 89 10 a 11 .2 16 b 18 .0 15 b 16 .1 41 46 .1 18 /8 /2 01 5 99 4a 4. 0 21 b 21 .2 1a 1. 0 26 26 .3 30 /8 /2 01 5 81 16 a 19 .8 13 a 16 .0 6b 7. 4 35 43 .2 9/ 9/ 20 15 57 11 a 19 .3 8b 14 .0 3c 5. 3 22 38 .6 25 /9 /2 01 5 21 1a 4. 8 11 b 52 .4 1a 4. 8 13 61 .9 20 /1 0/ 20 15 30 7a 23 .3 0b 0. 0 1b 3. 3 8 26 .7 11 /1 1/ 20 15 50 1a 2. 0 2a 4. 0 0a 0. 0 3 6. 0 9/ 12 /2 01 5 30 0 0. 0 0 0. 0 0 0. 0 0 0. 0 d at a w ith s im ila r le tt er s ar e no t s ig ni fic an tly d iff er en t ( p ≤ 0. 05 ) b et w ee n th e nu m be r of le av es w ith c ha sm ot he ci a on u pp er , u nd er a nd b ot h si de s fo r ea ch d at e. 5 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator at kanawat 1 and kanawat 2, 2 months after the first observation of the symptoms on the leaves. chasmothecia were formed on the upper or/and the under leaf surfaces. the highest percentage of infected leaves, on which chasmothecia were formed, reached 60% and 61.9 on 30 july and 25 september, respectively, with average of 38.7% from 20 july to 11 november. chasmothecia were formed on the upper surface in 12.5% of infected leaves, with highest occurrence of 23.3% on 20 august, and they were formed on the under surface in 18.4% of infected leaves, with highest occurrence of 52.4% on 25 september (tab. 2). in contrast, it was noted that the percentage of leaves, on which chasmothecia were formed on both sides, was relatively low with an average of 7.5% (fig. 1). variation in number of chasmothecia the number of chasmothecia formed on the leaves at the end of the season was estimated just in vineyard no. 1 (kanawat 1st) in 2014 and 2015, and vineyard no. 4 (kanawat 2nd) in 2015, but the numbers of chasmothecia on both sides of infected leaves were also estimated only in 2015. tab. 3 shows that formation of chasmothecia was more frequent on the leaf under side with an average of 0.6 / leaf than on the leaf upper side (0.4 / leaf ). the number of chasmothecia formed on both sides together was relatively low with an average of 0.2 / leaf. chasmothecia were produced on leaves from july to november. their number increased slowly between july and september, and it highly increased from september to the end of october, then it had a sudden reduction at the beginning of november . no new formation of chasmothecia was observed in december (fig. 2). the results showed also that the number of chasmothecia was variable between vineyards and years. in 2015, their number ranged between 0.2 (vineyard no. 4) to 2.6 (vineyard no. 1) per leaf (fig. 3).the first detection of chasmothecia on leaves was at the beginning of july in two surveyed vineyards and in both years of survey, but the number of chasmothecia was highly variable between years in the same vineyard (kanawat 1st). it reached 390 / 100 leaves in october 2014, but it was about 260 in october 2015. fig. 4 shows the chasmothecia average number in each month. the initial development of chasmothecia was observed on powdery mildew infected leaves on 20 july. most of them were immature (61.1%) with cream to yellow color, and 38.9% were semi-mature with brown color (tab. 4). yellow and brown chasmothecia had been observed until the beginning of october. their number quickly increased from mid-july to mid-august, while the amount of dark, mature chasmothecia began to increase rapidly after the middle of october (fig. 5). number of asci and ascospores in chasmothecia microscopic examination showed that chasmothecia were 85–180 μm in diameter. appendages, 7–12 in number, hyaline and sometimes brown at the basal half, sometimes not coiled, 140–300 μm in length, 2–3 μm in width. asci 1–5 (usually three) in an ascoma, 30–35 × 45–55 µm, and 65.6% of them were empty. ascospores 1–4 (usually three) in an ascus, 10–25 × 10–15 μm (tab. 5). during winter, the number of chasmothecia on the infected leaves decreased slowly, and their viability decreased suddenly between december and february (fig. 6). in march, at the time of bud break, chasmothecia were still present in small numbers on the leaves, but they had an average viability of 1.2% and 0.17% in april. fig. 1 number of grapevine leaves on which chasmothecia were formed from july to december 2015. 6 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator ta b. 3 v ar ia tio n in n um be r of c ha sm ot he ci a on u pp er o r/ a nd u nd er le af s ur fa ce s at k an aw at 1 st fr om ju ly to d ec em be r 20 15 . d at e n um be r of e xam in ed le av es n o. o f c ha sm oth ec ia fo rm ed on ly o n th e up pe r su rf ac e a ve ra ge o f ch as m ot he ci a n o. / le af u pp er su rf ac e n o. o f c ha sm oth ec ia fo rm ed on ly o n th e un de r su rf ac e a ve ra ge o f ch as m ot he ci a n o. / le af u nd er su rf ac e n o. o f c ha sm oth ec ia fo rm ed on b ot h su rf ac es a ve ra ge o f c ha sm ot he ci a n o. / bo th le af s id es to ta l n um be r of p ro du ce d ch as m ot he ci a a ve ra ge o f ch as m ot he ci a n o. / le af 8/ 7/ 20 15 20 0 0. 00 0 0. 00 0 0. 00 0 0. 00 20 /7 /2 01 5 10 6a 0. 60 3b 0. 30 9c 0. 90 18 1. 80 30 /7 /2 01 5 50 24 a 0. 48 20 ab 0. 40 15 b 0. 30 59 1. 18 6/ 8/ 20 15 89 40 a 0. 45 35 a 0. 39 18 b 0. 20 93 1. 04 18 /8 /2 01 5 99 39 a 0. 39 53 a 0. 54 14 b 0. 14 10 6 1. 07 30 /8 /2 01 5 81 32 a 0. 40 35 a 0. 43 25 a 0. 31 92 1. 14 9/ 9/ 20 15 57 36 a 0. 63 19 b 0. 33 11 c 0. 19 66 1. 16 25 /9 /2 01 5 21 8a 0. 38 43 b 2. 05 2c 0. 10 53 2. 52 20 /1 0/ 20 15 30 24 a 0. 80 51 b 1. 70 3c 0. 10 78 2. 60 11 /1 1/ 20 15 50 1a 0. 02 1a 0. 02 0a 0. 00 2 0. 04 9/ 12 /2 01 5 30 0 0. 00 0 0. 00 0 0. 00 0 0. 00 a ve ra ge 19 .0 9a 0. 38 23 .6 4a 0. 56 8. 82 b 0. 20 51 .5 5 1. 14 d at a w ith s im ila r le tt er s ar e no t s ig ni fic an tly d iff er en t ( p ≤ 0. 05 ) b et w ee n th e nu m be r of c ha sm ot he ci a on u pp er , u nd er a nd b ot h si de s fo r ea ch d at e. 7 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator fig. 2 variation in number of chasmothecia on upper or/and under leaf surfaces at kanawat 1st from july to december 2015. data with similar letters are not significantly different (p ≤ 0.05) between the number of chasmothecia on upper, under and both sides of leaves at the end of the growing season (october to november). fig. 3 number of chasmothecia observed on 100 leaves in vineyards no. 1 (kanawat 1st) and no. 4 (kanawat 2) from july to december 2015. numbers with similar letters are not significantly different (p ≤ 0.05) between observations in the same vineyard and between vineyards. fig. 4 the number of chasmothecia observed on 100 leaves in vineyard no. 1 (kanawat 1st) from july to november of 2014 and 2015. the number of chasmothecia was calculated as the average of observations made in each month. data with similar letters are not significantly different (p ≤ 0.05) between months in the same year and between years. 8 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator tab. 4 development of chasmothecia of erysiphe necator on leaves in kanawat 1st site in 2015. the percentages of young (cream or yellow), semi-mature (brown), and mature (dark brown to black) chasmothecia on infected leaves were determined. date chasmothecia color (%) appendage (%) cream/beige or yellow orange/brown black/dark brown absent initiation present 8/7/2015 0.0 0.0 0.0 0.0 0.0 0.0 20/7/2015 61.1 38.9 0.0 100.0 0.0 0.0 30/7/2015 81.4 13.6 5.0 55.9 0.0 44.1 6/8/2015 42.8 48.4 8.8 78.1 8.1 13.8 18/8/2015 30.2 55.7 14.1 76.5 13.2 10.4 30/8/2015 42.1 35.0 22.9 0.8 80.4 13.1 9/9/2015 39.3 40.8 19.9 66.7 24.2 9.1 25/9/2015 39.0 21.9 39.1 37.7 5.7 56.6 20/10/2015 18.5 7.8 73.6 57.1 14.3 28.6 11/11/2015 0.0 0.5 99.6 0.0 0.0 100.0 fig. 5 the percentages of young (cream or yellow), semi-mature (brown), and mature (dark brown to black) chasmothecia observed on infected leaves from july to november 2015. tab. 5 number of asci and ascospores in chasmothecia of erysiphe necator. location number of examined leaves number of examined chasmothecia number of asci / ascocarp number of ascospores/ascus 1 2 3 4 5 empty 1 2 3 4 kanawat 1st 100 90 1 2 20 6 2 59 0 6 15 4 salakad 78 58 1 10 15 2 2 28 2 12 28 2 9 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator discussion powdery mildew caused by e. necator is an important disease of grapevine in swieda, southern syria. the initial development of chasmothecia on infected leaves was observed at the second half of july in 45.5% of studied vineyards. this result is in accordance with those of many previous studies. in new york state, chasmothecia are produced from late july [4]. in germany, the first yellow fruiting bodies were detected as early as mid-july to mid-august, and the ascocarp formation stops at the beginning of october [2]. in contrast, in australia chasmothecia are formed during late april and early may [14]. in south africa, chasmothecia were also observed during april to may on severely infected leaves [15]. chasmothecia were formed on 38.7% of leaves in average, and they occurred in small number on leaves (1.1 / leaf ), and most of them were immature. these results are in according with those of halleen and holz [15] who showed that the number of chasmothecia formed on leaves were 1–10 per leaf. no ascospores were trapped by the spore traps, nor found on the bark in spring at the beginning of the season, although they have been observed on the leaves in fall. in contrast, holb and füzi [16] showed that ascospores were trapped from early april until end june, where 6.6% of the total ascospores were caught between the initiation of sampling in april and bud break, 62.2% from bud break to bloom, and 31.2% between bloom and the conclusion of sampling at the end of june. in winter and at the beginning of spring, chasmothecia were still present on leaves at a small number and with a very low viability. it seems that the viability of chasmothecia is strongly dependent on the environmental conditions. corties et al. [10] showed that the favorable climate of southern italy allowed the survival of the leaf litter until bloom, and leaves seemed to be a potential efficient substrate for overwintering chasmothecia. it seems also that the high temperatures have detrimental effects on the viability of chasmothecia [12]. according to these results, the low viability of chasmothecia may be explained by the high temperature at the beginning of season in southern syria. all these results confirm those of our previous study that the causal agent of grapevine powdery mildew (e. necator) overwinters as mycelium in dormant buds of the grapevines, and the ascospores did not had any role in the initiation of the disease in spring (unpublished data). the number of chasmothecia formed on leaves was highly variable between vineyards and years, these results are in according to those of hajjeh et al. [12]. the occurrence of e. necator chasmothecia were lower in 2015 than they were in 2014. previous study showed that the flag shoots and the disease severity were also more frequently in 2014 than they were in 2015 because the vineyards had been affected by frost in early spring 2015 when the temperature reached −16°c, which led to the death of most of the branches. spotts and chen [17] demonstrated that infected buds infected by powdery mildew were more susceptible to freezing injury and death than healthy buds. when temperatures dropped below −22°c, infected buds were likely to die whereas healthy buds could survive to temperatures dropping to −26°c. through the above, it seems that there is a very high correlation between the severity of powdery mildew infections on leaves in the vineyards and the number of chasmothecia on leaf area [5,7,18–20]. conclusion although e. necator chasmothecia are formed on the infected leaves, it seems that they may not have a role as a primary inoculum of powdery mildew of grapevine at the beginning of season in southern syria where this disease is very important, especially on ‘balady’ and ‘black’ grapevine cultivars. fig. 6 viability of chasmothecia on leaves during winter 2015. numbers indicate the surveyed vineyards. 10 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator acknowledgments the authors thank the team of plant protection laboratory (ministry of agriculture) in sweida for their help. references 1. gadoury dm, seem rc, pearson rc, wilcox wf. effects of powdery mildew on vine growth, yield, and quality of concord grapes. plant dis. 2001;85:137–140. http://dx.doi.org/10.1094/pdis.2001.85.2.137 2. hil gk, baumberger i, spies s. studies on the occurrence of the chasmothecia of uncinula necator (schw.) burr. in two vine growing areas of germany. viticulture and enology science. 1995;50:3–8. 3. fathi h, karbalaei h. study of biology and epidemiology of uncinula necator caused powdery mildew disease. technical journal of engineering and applied sciences. 2012;2:56–61. 4. moyer mm, gadoury dm, wilcox wf, seem rc. release of erysiphe necator ascospores and impact of early season disease pressure on vitis vinifera fruit infection. am j enol vitic. 2014;65:315–324. http://dx.doi.org/10.5344/ajev.2014.13111 5. gadoury dm, pearson rc. initiation, development, dispersal, and survival of chasmothecia of uncinula necator in new york vineyards. phytopathology. 1988;78:1413–1421. http://dx.doi.org/10.1094/phyto-78-1413 6. gordon cc. a reinterpretation of the ontogeny of the ascocarp of species of the erysiphaceae. am j bot. 1966;53:652–662. http://dx.doi.org/10.2307/2439741 7. legler se, caffi t, rossi v. a model for the development of erysiphe necator chasmothecia in vineyards. plant pathol. 2014;63(4):911–921. http://dx.doi.org/10.1111/ppa.12145 8. gadoury dm, cadle-davidson l, wilcox wf, dry ib, seem rc, milgroom mg. grapevine powdery mildew (erysiphe necator): a fascinating system for the study of the biology, ecology, and epidemiology of an obligate biotroph. mol plant pathol. 2012;13:1– 16. http://dx.doi.org/10.1111/j.1364-3703.2011.00728.x 9. pearson rc, gartel w. occurrence of hyphae of uncinula necator in buds of grapevine. plant dis. 1985;69:149–151. http://dx.doi.org/10.1094/pd-69-149 10. cortesi p, bisiach m, ricciolini m, gadoury dm. chasmothecia of uncinula necator – an additional source of inoculum in italian vineyards. plant dis. 1997;81:922–926. http://dx.doi.org/10.1094/pdis.1997.81.8.922 11. magarey pm, gadoury dm, emmett rw, biggins lt, clarke k, wachtel mp, et al. chasmothecia of uncinula necator in australia. viticulture and enology science. 1997;52:210–218. 12. hajjeh h, miazzi m, faretra f. overwintering of erysiphe necator schw. in southern italy. j plant pathol. 2008;90:323–330. 13. widholm jm. the use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. stain technol. 1972;47:189–193. http://dx.doi.org/10.3109/10520297209116483 14. wicks tj, magarey p, emmett rw. first report of uncinula necator cleistotbecia on grapevines in australia. plant dis. 1985;69:727. 15. halleen p, holz g. chasmothecia and flag shoots: sources of primary inoculum for grape powdery mildew in the western cape province, south africa. south african journal for enology and viticulture. 2000;21:66–70. 16. holb ij, füzi i. monitoring of ascospore density of erysiphe necator in the air in relation to weather factors and powdery mildew development. eur j plant pathol. 2016;144:751– 762. http://dx.doi.org/10.1007/s10658-015-0823-4 17. spotts ra, chen pm. cold hardiness and temperature responses of healthy and mildew infected terminal buds of apple during dormancy. phytopathology. 1984;7:542–544. http://dx.doi.org/10.1094/phyto-74-542 18. angeli d, pellegrini e, pertot i. occurrence of erysiphe necator chasmothecia and their natural parasitism by ampelomyces squisqualis. phytopathology. 2009;99:704–710. http://dx.doi.org/10.1094/phyto-99-6-0704 19. pearson re. current research on grape fungal diseases and their control in new york. australian and new zealand wine industry. 1990;5:206–209. http://dx.doi.org/10.1094/pdis.2001.85.2.137 http://dx.doi.org/10.5344/ajev.2014.13111 http://dx.doi.org/10.1094/phyto-78-1413 http://dx.doi.org/10.2307/2439741 http://dx.doi.org/10.1111/ppa.12145 http://dx.doi.org/10.1111/j.1364-3703.2011.00728.x http://dx.doi.org/10.1094/pd-69-149 http://dx.doi.org/10.1094/pdis.1997.81.8.922 http://dx.doi.org/10.3109/10520297209116483 http://dx.doi.org/10.1007/s10658-015-0823-4 http://dx.doi.org/10.1094/phyto-74-542 http://dx.doi.org/10.1094/phyto-99-6-0704 11 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1088 alimad et al. / chasmothecia development of erysiphe necator 20. cortesi p, gadoury dm, seem rc, pearson rc. distribution and retention of chasmothecia of uncinula necator on the bark of grapevines. plant dis. 1995;79:15–19. http://dx.doi.org/10.1094/pd-79-0015 http://dx.doi.org/10.1094/pd-79-0015 abstract introduction material and methods chasmothecia on leaves and branches trapping the released fungal spores statistical analysis results development of e. necator chasmothecia variation in number of chasmothecia number of asci and ascospores in chasmothecia discussion conclusion 2016-12-28t12:51:39+0000 piotr otręba 2014-11-20t23:19:58+0000 polish botanical society 2014-11-20t23:21:42+0000 polish botanical society 2014-11-20t23:20:24+0000 polish botanical society new and interesting species of lichens from xerothermic habitats in nw poland 1 of 12published by polish botanical society acta mycologica original research paper new and interesting species of lichens from xerothermic habitats in nw poland anetta wieczorek1,2*, andrzej łysko3, jurga motiejŭnaite4 1 department of ecology, university of szczecin, wąska 13, 71-412 szczecin, poland 2 center for molecular biology and biotechnology, faculty of biology, university of szczecin, wąska 13, 71-415 szczecin, poland 3 department of environmental protection and management, western pomeranian university of technology, słowackiego 17, 71-43 szczecin, poland 4 laboratory of mycology, nature research center, institute of botany, žaliųjų ežerų 49, 2021 vilnius, lithuania * corresponding author. email: anetta.wieczorek@usz.edu.pl abstract this paper presents data on the occurrence of lichens in xerothermic grasslands, representing a great mycological peculiarity of the nw part of pomerania, poland. the 12 examined specimens of six species originated from fieldwork carried out in 2011–2014 in the nature reserves brodogóry, stary przylep, bielinek, wrzosowiska cedyńskie, prof. adam wodziczko nature reserve in the wolin national park, and an old chalk excavation site on wolin island. within the study sites, four lichen species were recorded as a new to western pomerania: agonimia gelatinosa, collema cristatum, dermatocarpon luridum, and leptogium subtile. the other two species, collema auriforme and c. flaccidum, are rarely observed in the studied region. keywords biodiversity; lichens; nature reserves; nw poland introduction the great peculiarity of nw pomerania are a xerothermic grasslands. they develop in specific climatic conditions only, particularly in the areas where high soil and nearsurface air layer temperatures occur periodically. soils overgrown with xerothermic plants must be reasonably fertile and rich in calcium carbonate. such conditions create favorable habitats for development of xerothermophilous and calciphilous lichens [1]. in poland, calciphilous lichens usually occur in the mountains and uplands in limestone areas [2,3]. in lowlands, this group of lichens is rarely recorded in natural ranges of xerothermic grasslands of the lower vistula, odra, and warta rivers [4–7]. xerothermic grasslands of nw poland are found in morainic areas, characterized by varied relief. the vegetation is not dense, composed mostly of patches of the feather-grass community potentillo-stipetum capillatae. because of its loose structure and specific soil conditions, the community also includes many plants typical of sandy grasslands of the class koelerio glaucae-corynephoretea canescentis as well as thermophilous and heliophilous species of the class festuco-brometea [8,9] (and also unpublished documentation by dr tadeusz głazek from 1971 “rezerwat florystyczno-dydaktyczny ‘stary przylep’ ”). many lichens are adapted to adverse environmental conditions. they may colonize both natural and man-made substrata (such as concrete and bricks), and are often found in anthropogenic habitats, like wooden constructions, mechanically disturbed soils, worked stone surfaces and quarries. xerothermic lichens in northern poland are reported with increasing frequency in anthropogenic habitats, such as gravel pits, mine workings or quarries [10–12]. such moderate anthropogenic pressure creates new habitats for lichens and may significantly increase their diversity. this paper presents new localities of lichens that were recorded in xerothermic habitats in nw poland, developed partly as a result of human activity. doi: 10.5586/am.1097 publication history received: 2016-04-27 accepted: 2017-01-13 published: 2017-07-28 this article was originally submitted to and processed by the acta agrobotanica (another journal of the polish botanical society) editors. upon consent of the authors and the editors of both journals, it was eventually published in acta mycologica. handling editor piotr sugier (acta agrobotanica), faculty of biology and biotechnology, maria curieskłodowska university in lublin, poland authors’ contributions aw: research idea, coordination, and conduct, manuscript preparation; jm: manuscript preparation, reviewed drafts of the paper; ał: prepared figures and tables, reviewed drafts of the paper funding the research has been conducted on the authors own expenses. competing interests no competing interests have been declared. copyright notice © the author(s) 2017. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation wieczorek a, łysko a, motiejŭnaite j. new and interesting species of lichens from xerothermic habitats in nw poland. acta mycol. 2017;52(1):1097. https://doi. org/10.5586/am.1097 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:anetta.wieczorek%40usz.edu.pl?subject=new%20and%20interesting%20species%20of%20lichens%20from%20xerothermic%20habitats%20in%20nw%20poland https://doi.org/10.5586/am.1097 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1097 https://doi.org/10.5586/am.1097 2 of 12© the author(s) 2017 published by polish botanical society acta mycol 52(1):1097 wieczorek et al. / interesting lichens in nw poland material and methods the presented list of records is derived from the material collected during fieldwork in 2011–2014. the study included the following study sites: brodogóry nature reserve (b), stary przylep nature reserve (sp), prof. adam wodziczko nature reserve (aw) in the wolin national park, an old chalk (oc) excavation site on wolin island, bielinek nature reserve (bi), and wrzosowiska cedyńskie nature reserve (wc) (fig. 1). in total, 12 specimens of six species were collected, now deposited in the lichen herbarium (szub-l) of the department of ecology and environmental protection at the university of szczecin. morphological and anatomical observations were made using standard microscopic techniques, under an inverted fluorescence microscope (zeiss axio observer a1) and a light microscope (zeiss axio scope a1). fig. 1 distribution map of the study sites, with numbers of the listed species found. aw – prof. adam wodziczko nature reserve in the wolin national park; oc – old chalk excavation site on wolin island; b – brodogóry nature reserve; sp – stary przylep nature reserve; wc – wrzosowiska cedyńskie nature reserve; bi – bielinek nature reserve; 1 – agonima gelatinosa; 2 – collema auriforme; 3 – c. cristatum; 4 – c. flacidium; 5 – dermatocarpon luridum; 6 – leptogium subtile. 3 of 12© the author(s) 2017 published by polish botanical society acta mycol 52(1):1097 wieczorek et al. / interesting lichens in nw poland results: list of recorded species agonima gelatinosa (ach.) brand & diederich (fig. 2a) terricolous (found on calcareous soil), muscicolous and also on plant debris. new to nw poland. this rare species in poland is known from the gorce mts [13,14], kujawy region [7,15,16], przemyśl town [17], sudety mts [18], and tatry mts [19]. specimens examined. old chalk excavation site on wolin island (n 53°53'08.1" / e 14°27'11.7") on soil, leg. anetta wieczorek, andrzej łysko, szub-l 3008; wrzosowis ka cedyńskie nature reserve (n 52°51'16.1" / e 14°10'27.3"), on soil, leg. anetta wieczorek, andrzej łysko, szub-l 3009; bielinek nature reserve (n 52°55'24.7" / e 14°10'18.1"), on soil, leg. anetta wieczorek, andrzej łysko, szub-l 3007; brodogóry nature reserve (n 53°21'32.7" / e 14°93'08"), on soil, leg. anetta wieczorek, andrzej łysko, szub-l 3010. collema auriforme (with.) coppins & j. r. laundon (fig. 2b) in pomerania recorded earlier at one site in bory tucholskie forest [20,21]. it is also reported from southern poland, e.g., the beskid mały mts [22], beskid sądecki mts [23], beskid śląski mts [24], beskid wyspowy mts and beskid żywiecki mts [25], stołowe mts [26], kielce town [27], kraków city [28], pieniny mts [29], spiskie foothills [30], rożnowskie foothills [31], sudety mts [32–34], tatry mts [35–38], tarnów town [39], near warsaw [40], and kraków-częstochowa upland [41]. specimens examined. old chalk excavation site on wolin island (n 53°53'08.1" / e 14°27'11.7"), found on mosses, calcareous soil, in moist and shaded conditions, in anthropogenic habitats. it develops characteristic isidia in brownish-green lobes, leg. anetta wieczorek, andrzej łysko, szub-l 3006; bielinek nature reserve (n 52°55'24.7" / e 14°10'18.1"), on calcareous soil in natural habitats, leg. anetta wieczorek, andrzej łysko, szub-l 3004; stary przylep nature reserve (n 53°19'08" / e 14°99'41"), on calcareous soil, leg. anetta wieczorek, andrzej łysko, szub-l 2996. collema cristatum (l.) weber in f. h. wigg. (fig. 2c) within poland reported previously only from the southern part of the country, e.g., the beskidy zachodnie mts [42], beskid sądecki mts [23], vistula valley [43], świętokrzyskie mts [44], kielce town [27], kraków city [28], chęciny region [45], pieniny mts [3,29], śląsk opolski town [46], tatry mts [37,47,48], kraków-częstochowa upland [41], and wieluńska upland [49]. specimens examined. prof. adam wodziczko nature reserve in the wolin national park (n 53°51'50.0" / e 14°27'06.0"), on calcareous soil on sun-exposed sites in natural habitats, leg. anetta wieczorek, andrzej łysko, szub-l 3003; old chalk excavation site on wolin island (n 53°53'08.1" / e 14°27'11.7"), on calcareous soil, in exposed conditions, in artificial (excavation) habitats, leg. anetta wieczorek, andrzej łysko, szub-l 3002. collema flaccidum (ach.) ach. (fig. 2d) in poland earlier known from several localities in the north of the country and several in the south [50]. reported in the north of kaszubskie coastland [20,51,52], suwałki landscape park [53], and near białowieża national park [54]. in the south of the country reported from the bieszczady mts [55,56], beskid niski mts [57], beskid mały mts [22], beskid sądecki mts [23,58,59], beskid średni mts [60], beskid wyspowy mts and beskid żywiecki mts [61], gorce mts [62], stołowe mts [26], sudety mts [63], and tatry mts [35,37,64,65]. 4 of 12© the author(s) 2017 published by polish botanical society acta mycol 52(1):1097 wieczorek et al. / interesting lichens in nw poland fig. 2 thalli of xerothermic lichens. 1 – agonima gelatinosa; 2 – collema auriforme; 3 – c. cristatum; 4 – c. flacidium; 5 – dermatocarpon luridum; 6 – leptogium subtile. 5 of 12© the author(s) 2017 published by polish botanical society acta mycol 52(1):1097 wieczorek et al. / interesting lichens in nw poland specimens examined. wrzosowiska cedyńskie nature reserve (n 52°51'16.1" / e 14°10'27.3"), on damp rock, among mosses in open habitats, on the edge of the reserve, leg. anetta wieczorek, andrzej łysko, szub-l 3001. dermatocarpon luridum (with.) j. r. laundon (fig. 2e) the record from the old chalk excavation site is the first record from northern poland. previously reported from only five localities in southern poland, e.g., the bieszczady mts [55,56], beskid wyspowy mts and beskid żywiecki mts [61], lower silesia [66], sudety mts [63], and tatry mts [37,38,65,67–70]. specimens examined. old chalk excavation site on wolin island (n 53°53'08.1" / e 14°27'11.7"), grows on exposed calcareous soil around the margins of water bodies, leg. anetta wieczorek, andrzej łysko, szub-l 3011. leptogium subtile (schrad.) torss. (fig. 2f ) new to northern poland. previously reported from only two localities in southern poland: bielska plain [71] and the gorce mts [62]. specimens examined. old chalk excavation site on wolin island (n 53°53'08.1" / e 14°27'11.7"), on debarked wood and mosses in humid places, leg. anetta wieczorek, andrzej łysko, szub-l 2998. discussion in the studied localities, xerothermic epigeic lichens were found on calcareous soil, humus, and decaying remains of plants as well as on rock-waste. calciphilous lichens were not homogeneous with respect to specific habitat requirements. here, a common factor was only the chemical nature of substratum (calcium carbonate). other factors differed, e.g., the degree of sun exposure or substratum moisture [72]. in total, six new species of lichenized fungi were observed: one in the prof. adam wodziczko nature reserve in the wolin national park, five in the old chalk excavation site on wolin island, one in the brodogóry nature reserve, one in the stary przylep nature reserve, two in the wrzosowiska cedyńskie nature reserve, and two in the bielinek nature reserve (fig. 1). the species are either new or rare in western pomerania. all the collected lichens were found in isolated localities, in small populations. their thalli were healthy, with well-developed reproductive organs, and did not show any symptoms of decline. four of the recorded lichens were new to nw pomerania: agonimia gelatinosa, collema cristatum, dermatocarpon luridum, and leptogium subtile (fig. 2). they were found together in the old chalk excavation site located in the buffer zone of the wolin national park. lichens of xerothermic habitats are usually inconspicuous and rarely observed. they are described chiefly during specialized lichenological explorations. currently, their distribution in europe is well-studied. they are recorded in several climatic zones, from the boreal to the mediterranean zone [73], but most frequently in se and s parts of europe with a continental climate. in other parts of the continent, with a more humid climate, isolated patches of steppe vegetation are found on particularly warm and dry sites [74]. the patches occupy calcium-rich, steep riverbanks, ravines, or calcareous rocks. in central europe, xerothermic lichens are rare or very rare, particularly in the lowlands [75–78] (tab. 1). the small size of the patches and the increasing eutrophication of adjacent areas are important threats to those populations. the presented records are some of the northernmost localities of xerothermic lichens, which are particularly important for the dynamics of those taxa. 6 of 12© the author(s) 2017 published by polish botanical society acta mycol 52(1):1097 wieczorek et al. / interesting lichens in nw poland tab. 1 other european records of the lichen species reported in this study. species countries agonima gelatinosa (ach.) m. brand & diederich estonia [79], netherlands [78], poland [80] collema auriforme (with.) coppins & j. r. laundon albania [81], austria [82], bosnia and herzegovina [83], bulgaria [84], croatia [85], france [86], germany [87], greece [88,89], iceland [90], macedonia [91], netherlands [78], poland [80], slovakia [92], slovenia [93,94], spain [95,96], switzerland [97–101], turkey [102–106] collema cristatum (l.) f. h. wigg. austria [82], bosnia and herzegovina [83], bulgaria [84], croatia [85], estonia [79], germany [87], italy [107,108], ireland [109], lithuania [110], poland [80], russia [111], slovenia [94,107], spain [96,112], switzerland [101,113,114], turkey [115–123] collema flaccidum (ach.) ach. austria [82,124,125], bosnia and herzegovina [83], bulgaria [84], france [126], germany [88,127], greece [89], latvia [128], macedonia [91], poland [80], romania [129], serbia [130], switzerland [98,101] dermatocarpon luridum (with.) j. r. laundon bosnia and herzegovina [83], bulgaria [84], lithuania [131], estonia [132], germany [87], greece [89], poland [80], portugal [133], spain [134], 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(sensu stricto) and a. ostoyae (romagn.) herink) have fruiting bodies with a ring on stem and constitute armillaria mellea complex. the remaining ones: a. ectypa (fr.) lamoure and a. tabescens (scop.) emel are characterized by the lack of such a ring [3,4]. geographical distribution of the species of the genus armillaria in poland is quite diverse. for example, a. cepistipes, a. gallica and a. ostoyae, are encountered all over the country, whereas a. borealis was found only in its northern and central parts [3,5]. among the species extremely rarely noted in poland is marsh honey fungus – armillaria ectypa, which has been under strict protection since 2014 [6]. armillaria ectypa is ranked as threatened and rare not only in poland but also in many other european countries [7,8] it is one of 33 fungi proposed for inclusion into the bern convention [9,10]. the purpose of this paper is to present the basidiocarp morphology, ecological notes and distribution of a. ectypa in poland. material and methods the description of macroscopic and microscopic features is based on fresh and dried specimens, collected in żuromino (the kaszuby lakeland). the microscopic structures doi: 10.5586/am.1064 publication history received: 2015-07-23 accepted: 2015-07-28 published: 2015-08-05 handling editor tomasz leski, institute of dendrology of the polish academy of sciences, poland funding the study was financed by the university of szczecin as part of individual research grants. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation stasińska m. armillaria ectypa, a rare fungus of mire in poland. acta mycol. 2015;50(1):1064. http://dx.doi.org/10.5586/ am.1064 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:stasinsk%40univ.szczecin.pl?subject=armillaria%20ectypa%2c%20a%20rare%20fungus%20of%20mire%20in%20poland http://dx.doi.org/10.5586/am.1064 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1064 http://dx.doi.org/10.5586/am.1064 2 of 6© the author(s) 2015 published by polish botanical society acta mycol 50(1):1064 stasińska / armillaria ectypa, a rare fungus of mire in poland were observed and measured using olympus bx53 light microscope (lm) equipped with a olympus dp26 digital camera. basidia and spore measurements in the descriptions are based on 30–40 measurements. dimensions of the basidia and the spores are given as follows: (minimum value–)1st decile–9th decile(–maximum value). spore length to width ratios are reported as q. micrographs taken with a scanning electron microscope zeiss evo ls10 (sem) in the center for molecular biology and biotechnology, environmental testing laboratory, university of szczecin (poland). the specimens were identified by examining their macroscopic and microscopic features, using references by termorshuizen [11] and vesterholt [12]. distribution of species in poland (fig. 1) was mapped according to the atpol grid square system as used by wojewoda [13]. the general distribution of species and ecological notes are based on original material, supplemented by information from the literature. the fungal nomenclature and its synonyms are given according to index fungorum database [14]. the nomenclature of vascular plants follows mirek et al. [15] and that of plant communities follows matuszkiewicz [16]. the collection studied are deposited in the herbarium of the department of botany and nature conservation, szczecin university (szub), poland. results species description armillaria ectypa (fr.) lamoure, compt. rend. hebd. séanc. acad. sci., paris 260: 4562 (1965) – physalacriaceae, agaricales, agaricomycetidae, agaricomycetes, agaricomycotina, basidiomycota, fungi [1]. syn.: agaricus ectypus fr., armillaria ectypa (fr.) herink; armillariella ectypa (fr.) singer; clitocybe ectypa (fr.) gillet; omphalia ectypa (fr.) quél.; for other synonymies see index fungorum [14]. pileus 25–60(100) mm in diameter, convex to plano-convex, later explanate, sometime with small obtuse umbo or depressed center with age, with slightly curved or waved margin, hygrophanous, brown, yellowish brown or ochraceous brown to pale cream-brown at margin, with dark brown scales at center, margin translucently striated (fig. 2); lamellae decurrent, rather narrow, first whitish then pale cream, ochraceous to pinkish; stipe 50–100 × 5–11(13) mm, cylindrical to subclavate, slightly thickened at base, without ring, fibrillose, of the same color as the pileus; context thin, white; without rhizomorphs; spore print whitish or creamish; smell fungoid; taste mild. spores (6.5)7.0–8.5(9.0) × (5.0)5.5–6.5(7.0) µm, q = 1.2–1.5, broadly ellipsoid to ellipsoid; basidia 30–40 × 7.0–8.0(9.0) µm, clavate, mostly with 4 sterigmata (fig. 3a,b). fig. 1 distribution of armillaria ectypa in poland, in atpol grid square systems [13]. fig. 2 fruit body of armillaria ectypa recorded in żuromino (photo: m. stasińska). 3 of 6© the author(s) 2015 published by polish botanical society acta mycol 50(1):1064 stasińska / armillaria ectypa, a rare fungus of mire in poland habitat and distribution armillaria ectypa is a saprotrophic species, growing on wet ground, peat, usually among sphagnum spp. and herbaceous plants (e.g., carex spp., drosera spp., eriophorum spp., phragmites australis) as well as on tufts of decaying sedges, e.g., carex rostrata. it occurs on waterlogged habitats, active raised bogs and alkaline fens [9] and aapa mires, transitional and low [11,12,17–19]. marsh honey fungus is an eurasian species of boreal-mountain range and possibly continental [19]. it is known in 14 european countries, i.e., austria, switzerland [10], the czech republic [8], denmark, sweden [12], finland [19], france [17], germany [20], great britain [18,21–23], the netherlands [11], russia [24] and slovakia [25]. in addition, it was reported from china [26] (cited in [27]), japan [28] and turkey [29]. in poland, a. ectypa was recorded first in leucobryo-pinetum w. mat. (1962) 1973, in the laski forest district [30]. the second site, discovered in september 2007, is located within the kaszuby lakeland, in the vicinity of żuromino [31]. armillaria ectypa was found on a small transitional bog, surrounded with a lagg zone filled with water, situated in a land depression among arable fields. fruiting bodies of marsh honey fungus were observed at a few places, singly or in small clusters, mainly on the margin of the peat bog, among sphagnum spp. (e.g., sph. cuspidatum and sph. fallax) and vascular plants (e.g., andromeda polifolia, carex lasiocarpa, c. rostrata, comarum palustre, drosera rotundifolia, eriophorum angustifolium, menyanthes trifoliata, oxycoccus palustris and viola palustris). the third stand of armillaria ectypa, is located in the wigry national park [32]. fruiting bodies were found in the community scheuchzerietalia palustris nordh. 1937, among sphagnum spp. and on the remnants of sphagnum mosses. localities in poland specimens indicated by asterisk (“*”) have not been examined by the author. bc-06 – południowobałtyckie lakelands: żuromino; transitional bog, among sphagnum spp. (e.g., sph. cuspidatum and sph. fallax) and vascular plants (e.g., carex rostrata, comarum palustre, drosera rotundifolia, eriophorum angustifolium and menyanthes trifoliata), 20 september 2007, leg. et det. m. stasińska, szub [31]. bg-10 – wschodniobałtyckie lakelands: wigry national park; in community of the order scheuchzerietalia palustris, rests of sphagnum mosses, among sphagnum spp., 30 september 2009, leg. et det. m. halama, mh-2009-0120 [32]*. ec-37 – środkowopolskie lowlands: laski (dobrygość) forest district, forest section no. 29h; leucobryo-pinetum molinietosum [30]*. fig. 3 armillaria ectypa. a,b spores and basidia seen by sem. 4 of 6© the author(s) 2015 published by polish botanical society acta mycol 50(1):1064 stasińska / armillaria ectypa, a rare fungus of mire in poland acknowledgments the author expresses gratitude to dr. marek halama (university of wrocław) for making his unpublished material available for publication in the present paper and dr. bożena białecka [center for molecular biology and biotechnology, environmental testing laboratory, university of szczecin (poland)] for taking sem pictures. the author thanks two anonymous reviewers for helpful suggestions on the manuscript. references 1. kirk pm, cannon pf, minter dw, stalpers ja, editors. dictionary of the fungi. 10th ed. wallingford: cabi; 2008. 2. wojewoda w. checklist of polish larger basidiomycetes. cracow: w. szafer institute of botany, polish academy of sciences; 2003. (biodiversity of poland; vol 7). 3. żółciak a. rozmieszczenie grzybów z rodzaju armillaria w polsce oraz ich rośliny żywicielskie. prace instytutu badawczego leśnictwa, seria a. 2003;3(956):7–22. 4. żółciak a. taksonomia i nomenklatura rodzaju armillaria (fr.: fr.) staude. prace instytutu badawczego leśnictwa, seria a. 2003;2(952):5–21. 5. żółciak a. opieńki. warszawa: centrum informacyjne lasów państwowych; 2005. 6. rozporządzenie ministra środowiska z dnia 9 października 2014 r. w sprawie ochrony gatunkowej grzybów. dz. u. z dnia 16 października 2014 r., poz. 1408. 7. wind p, pihl s, editors. the danish red list [internet]. the national environmental discussion and conclusions armillaria ectypa, has been recorded in poland only on three stands, although one of them, in leucobryo-pinetum [30], seems to be doubtful. this species is first of all encountered in very wet places, mostly on different kinds of peatland [11,12,17–19,22], therefore, it is unlikely to occur on such distinctly different stand as pine forest [30]. the other two stands, where it was recorded, are on transitional bogs, the habitats typical of this species [31,32]. according to ohenoja [19] a. ectypa is a good indicator of the mesotrophic mire habitats. apart from a high water availability and fertile microhabitat, a low nitrogen content in the soil is probably an essential factor determining the occurrence of a. ectypa [33]. this is probably why fruiting bodies of a. ectypa has not been found on the transitional bog near żuromino (despite the search in 2010 and 2012) strongly influenced by the great amount of biogenic compounds, including nitrogen from the surrounding arable lands. observed for many years all over the world, drainage and degradation of waterlogged areas, swamps and peatland [34–36] cause, that a. ectypa is a threatened species all over europe [9], and probably critically endangered since in some countries it has not been recorded for many years, e.g., in germany [37,38] and switzerland [39]. in many countries it is a rare species, recorded on a single or few stands [9]. only in finland it was found on over 30 stands [19]. in 11 out of 17 countries, where a. ectypa is known, it was included into the red list of fungi, e.g., in the czech republic (critically endangered cr [40]), denmark (endangered en [7]), great britain (endangered b [41]), germany [category 1 (critically endangered cr) [38]] and russia – tula region (category 2, [24]). the species is also included in the preliminary european fungal red list [33]. in poland, a. ectypa belongs to an extremely rare species as it can be seen from the number of recorded stands. moreover, it is a threatened taxon due to its occurrence in the habitats strongly saturated with water such as different peatlands, which have been undergoing fast and unfavorable alterations for many years [36]. therefore, taking into account the above reasons, marsh honey like some other taxons [42] is warrant being placed on the red list of macrofungi [43]. 5 of 6© the author(s) 2015 published by polish botanical society acta mycol 50(1):1064 stasińska / armillaria ectypa, a rare fungus of mire in poland research institute, aarhus university. 2004 [cited 2015 mar 30]; available from: http:// redlist.dmu.dk 8. antonín v, dvořák d. new, rare and lesser-known macromycetes in moravia (czech republic) – ix. acta musei moraviae, scientiae biologicae (brno). 2010;95(1):143–162. 9. dahlberg a, croneborg h, editors. the 33 threatened fungi in europe: complementary and revised information on candidates for listing in appendix 1 of the bern convention. uppsala: swedish species information center; 2003. 10. dahlberg a, croneborg h, editors. the 33 threatened fungi in europe: complementary and revised information on candidates for listing in appendix 1 of the bern convention. nature and environment. 2006;136:1–131. 11. termorshuizen aj. genus armillaria (fr.: fr.) staude. in: bas c, kuyper tw, noordeloos me, vellinga ec, editors. flora agaricina neerlandica: critical monographs on families of agarics and boleti occurring in the netherlands. rotterdam: balkema; 1995. p. 34–39. 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(biodiversity of poland; vol 1). 16. matuszkiewicz w. przewodnik do oznaczania zbiorowisk roślinnych polski. warszawa: pwn; 2001. 17. żółciak a, bouteville rj, tourvieille j, roeckel-drevet p, nicolas p, guillaumin jj. occurrence of armillaria ectypa (fr.) lamoure in peat bogs of the auvergne – the reproduction system of the species. cryptogam mycol. 1997;18(4):299–313. 18. ainsworth am. report on the marsh honey fungus armillaria ectypa, a uk bap species. english nature research reports. 2003;540:1–23. 19. ohenoja e. armillaria ectypa, a vulnerable indicator of mires. acta mycol. 2006;41(2):223– 228. http://dx.doi.org/10.5586/am.2006.024 20. krieglsteiner gj. verbreitungsatlas der grosspilze deutschlands (west). 1. ständerpilze. teil b: blätterpilze. stuttgart: e. ulmer gmbh & co.; 1991. 21. ainsworth am. searching for luminous mushrooms of the marsh fungus armillaria ectypa. field mycology. 2004;5(4):141–144. http://dx.doi.org/10.1016/s1468-1641(10)60279-4 22. wright m. update on armillaria ectypa at garron plateau, northern ireland. field mycology. 2007;8(2):41–43. http://dx.doi.org/10.1016/s1468-1641(10)60450-1 23. legon nw, henrici a, roberts pj, spooner bm, watling r. checklist of the british and irish basidiomycota, 4th update of the printed version published 2005 [internet]. 2009 [cited 2015 mar 30]; available from: http://www.basidiochecklist.info 24. shcherbakov av, editor. red data book of tula region: plants and fungi [internet]. 2010 [cited 2015 mar 30]; available from: http://www.wsl.ch/eccf/russian-redbook-fungi_ jan_2011.xls 25. škubla p. mycoflora slovaca. number of the copy 10. bratislava: mycelium edition; 2003. 26. zhang m. the fungi in the region of hengduan mountains. beijing: science press; 1996. 27. qin gf, zhao j, korhonen k. a study on intersterility groups of armillaria in china. mycologia. 2007;3:430–441. http://dx.doi.org/10.3852/mycologia.99.3.430 28. kudo s, nagasawa e. armillaria ectypa rediscovered in aomori prefecture, northern japan. reports of the tottori mycological institute. 2003;41:26–34. 29. sesli e, denchev cm. checklist of the myxomycetes, larger ascomycetes, and larger basidiomycetes in turkey. mycotaxon. 2008;106:65–67. 30. kowalski s. zbiorowiska grzybów leśnego środowiska glebowego wybranych drzewostanów sosnowych. prace komisji nauk rolniczych i komisji nauk leśnych. 1974;38:123–165. 31. stasińska m. macrofungi of raised and transitional bogs of pomerania. łódź: polish http://redlist.dmu.dk http://redlist.dmu.dk http://www.indexfungorum.org http://www.indexfungorum.org http://dx.doi.org/10.5586/am.2006.024 http://dx.doi.org/10.1016/s1468-1641(10)60279-4 http://dx.doi.org/10.1016/s1468-1641(10)60450-1 http://www.basidiochecklist.info http://www.wsl.ch/eccf/russian-redbook-fungi_jan_2011.xls http://www.wsl.ch/eccf/russian-redbook-fungi_jan_2011.xls http://dx.doi.org/10.3852/mycologia.99.3.430 6 of 6© the author(s) 2015 published by polish botanical society acta mycol 50(1):1064 stasińska / armillaria ectypa, a rare fungus of mire in poland botanical society; 2011. (monographiae botanicae; vol 101). http://dx.doi.org/10.5586/ mb.2011.001 32. halama m, romański m, krzysztofiak l, krzysztofiak a. chronione grzyby makroskopijne (macromycetes) wigierskiego parku narodowego. wigry. 2015 (in press). 33. the global fungal red list initiative [internet]. 2015 [cited 2015 mar 30]; available from: http://iucn.ekoo.se/iucn/species_view/326450 34. aapala k, lindholm t, heikkilä r. protected mires in finland. gunneria. 1995;70:205–220. 35. moen a. introduction: regionality and conservation of mires. gunneria. 1995;70:11–22. 36. herbich j, herbichowa m. szata roślinna torfowisk polski. in: ilnicki p, editor. torfowiska i torf. poznań: wyd. ar w poznaniu; 2002. p. 179–203. 37. kreisel h. rote liste der gefährdeten grosspilze mecklenburg-vorpommerns. die umweltministerin des landes mecklenburg-vorpommern. schwerin: goldschmidt druck gmbh; 1992. 38. karasch p, hahn c. rote liste gefahrdeter grospilze bayerns [internet]. 2009 [cited 2015 mar 30]; available from: http://www.lfu.bayern.de/natur/rote_liste_pilze/doc/roteliste_ grosspilze.pdf 39. senn-irlet b, bieri g, egli s. rote liste der gefahrdeten grosspilze der schweiz. umweltvollzug nr. 0718. bern: bafu, wsl; 2007. 40. holec j, beran m, editors. červený seznam hub (makromycetů) české republiky. praha: příroda; 2006. (vol 24). 41. evans s, henrici a, ing b. the red data list of threatened british fungi [internet]. 2006 [cited 2015 mar 30]; available from: http://www.britmycolsoc.org.uk/mycology/conservation/ red-data-list/ 42. friedrich s. the occurrence of amanita strobiliformis (paulet ex vittad.) bertill. in szczecin and its distribution in poland. acta mycol. 2013;48(1):123–131. http://dx.doi.org/10.5586/ am.2013.014 43. wojewoda w, ławrynowicz m. red list of the macrofungi in poland. in: mirek z, zarzycki k, wojewoda w, szeląg z, editors, red list of plants and fungi in poland. cracow: w. szafer institute of botany, polish academy of sciences; 2006. p. 53–70. http://dx.doi.org/10.5586/mb.2011.001 http://dx.doi.org/10.5586/mb.2011.001 http://iucn.ekoo.se/iucn/species_view/326450 http://www.lfu.bayern.de/natur/rote_liste_pilze/doc/roteliste_grosspilze.pdf http://www.lfu.bayern.de/natur/rote_liste_pilze/doc/roteliste_grosspilze.pdf http://www.britmycolsoc.org.uk/mycology/conservation/red-data-list/ http://www.britmycolsoc.org.uk/mycology/conservation/red-data-list/ http://dx.doi.org/10.5586/am.2013.014 http://dx.doi.org/10.5586/am.2013.014 abstract introduction material and methods results species description habitat and distribution localities in poland discussion and conclusions acknowledgments references 2015-08-03t19:35:13+0100 piotr otręba professor andrzej piotr grzywacz dr. h. c. mult. acta mycologica article id: 5611 doi: 10.5586/am.5611 publication history received: 2021-09-30 accepted: 2021-10-15 published: 2021-11-30 handling editor wojciech pusz; wojciech pusz; wrocław university of environmental and life sciences, poland; https://orcid.org/0000-00031531-2739 funding this work did not involve any funding. competing interests as is associate editor of acta mycologica. copyright notice © the author(s) 2021. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. memories and scientists professor andrzej piotr grzywacz dr. h. c. mult. andrzej szczepkowski * institute of forest sciences, warsaw university of life sciences – sggw, nowoursynowska 159, 02-776 warsaw, poland * to whom correspondence should be addressed. email: andrzej_szczepkowski@sggw.edu.pl professor andrzej p. grzywacz was born on july 10, 1943, in toruń, where he completed primary and secondary schooling. during the years 1961–1966, he studied at the faculty of forestry of the warsaw university of life sciences (sggw). his interest in mushrooms was evident since then through his choice of specialization and the title of his master’s thesis grzyby nadrzewne w leśnictwach wiartel i zielone (puszcza piska) [arboreal fungi in the wiartel and zielone forest subdistricts (puszcza piska forest)], which he carried out under the supervision of prof. jerzy ważny. aerwards, he worked in the state forests in the forest districts smolniki (now iława) and olek (now toruń). in 1972 he completed his doctoral studies (1969–1972) under the supervision of prof. jerzy ważny at the faculty of forestry, warsaw university of life sciences (sggw), and he defended his doctoral dissertation wpływ przemysłowych zanieczyszczeń powietrza na niektóre patogeniczne grzyby drzew leśnych [e influence of industrial air pollution on selected pathogenic fungi of forest trees]. in 1979, andrzej grzywacz was awarded the degree of doctor habilitatus in forest sciences on the basis of his output and the monograph entitled mechanizm odporności nasion pinus sylvestris l. na pasożytniczą zgorzel przedwschodową [mechanism of resistance of pinus sylvestris l. seeds to parasitic pre-emergence damping-off]. he started his journey as an associate professor in 1988 and was awarded the title of full professor in 1995. in 1998, he was elected as a corresponding member, and in 2007, as a full member of the polish academy of sciences. professor grzywacz was conferred the title of doctor honoris causa by the august cieszkowski university of agriculture (now poznań university of life sciences) (2008), the university of agriculture in kraków (2014), and the wrocław university of environmental and life sciences (2017). from 1969 until his retirement, prof. a. grzywacz worked at the faculty of forestry of the warsaw university of life sciences. from 1980 to 2012, he served as the head of the laboratory and later as the head of the department of mycology and forest phytopathology of the sggw, which was established on his initiative. he was vice acta mycologica / 2021 / volume 56 / article 5611 publisher: polish botanical society 1 https://doi.org/10.5586/am.5611 https://orcid.org/0000-0003-1531-2739 https://orcid.org/0000-0003-1531-2739 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0002-9778-9567 mailto:andrzej_szczepkowski@sggw.edu.pl szczepkowski / professor andrzej piotr grzywacz dr. h. c. mult. dean for teaching (1981–1984), dean of the faculty of forestry (1990–1993), and a vice rector for teaching and education at sggw (1984–1990). during the course of his extensive professional activity and thematically diversified scientific achievements, several areas that were of interest to him are as follows: forest pathology and mycology, nature protection, policy of the state within the fields of ecology and forestry, development of forest sciences, reform and improvement of forestry studies, education of the society within the field of forestry, and history of forestry. in the fields of phytopathology and mycology, he particularly dealt with the influence of industrial air pollution on forest tree pathogenic fungi, chemical control of seedling damping-off and other diseases in forest nurseries, the mechanism of seed resistance to parasitic pre-emergence damping-off, pathogenic fungi of transplanting stress in older trees, diseases of introduced conifers, provenance resistance to rhabdocline pseudotsugae of douglas fir, health of ash trees in urban conditions, ectomycorrhizal fungi for controlled mycorrhization of seedlings in container cultivation, biology and ecology of wood-decaying fungi in the forest and outside, decline of beech and alder stands, chemical wood protection, fungitoxicity of fungicides, legal protection of macrofungi species, monitoring of macrofungi, species diversity of fungi in forest ecosystems, health status of trees, and natural monuments. prof. a. grzywacz has about 500 publications, including 135 original papers published in prestigious polish (e.g., acta societatis botanicorum poloniae, acta mycologica, folia forestalia polonica, sylwan) and international (e.g., european journal of forest pathology) journals, scientific monographs and chapters in monographs, 20 books, articles in proceedings of symposia and scientific conferences, extensive review publications, articles in popular scientific and branch press, instruction articles for forest practice and numerous editions of scientific books and materials of scientific symposia, especially those organized by the polish forest society (ptl). he was the founder and principal investigator of 14 research projects commissioned by the scientific research committee (kbn), the state forests, the forest research institute, and participated in the implementation of seven research projects commissioned by the u.s. department of agriculture fg-pol-304, the polish academy of sciences, and the national fund for environmental protection and water management. prof. grzywacz has written over 110 expert opinions, projects, and studies for forest practice, nature conservation, protection of historical monuments, and the parliament office of analyses. furthermore, he has written over 50 prepublication reviews of books and scripts for various publishing houses and over 150 reviews of applications for research projects funded by the state committee for scientific research, ministry of science and higher education, and the polish science foundation. he has peer-reviewed many manuscripts for polish and foreign journals and provided his expert opinion on research documentation, applications for scientific prizes, dras of legislative acts for the ministry of the environment, plans for the protection of national parks, and other studies. prof. a. grzywacz has visited numerous scientific institutions in belarus, bulgaria, the czech republic, lithuania, mexico, germany, russia, slovakia, and hungary. he was a member of polish delegations at scientific congresses, such as the iii plant protection congress (munich), x world forestry congress (paris), international union of forest research organizations (iufro) 100th anniversary congress (berlin), and iufro congresses in tampere (finland), brisbane (australia), and seoul (south korea). prof. a. grzywacz has lectured (both full-time and part-time) postgraduate and doctoral students, not only at the faculty of forestry but also at the biology, environmental protection, tourism and recreation, wood technology (warsaw university of life sciences), and ethnography (on protection of antique wood) departments at the university of warsaw. he mainly taught forest pathology, nature protection, nature-forest education, and policy of the state within the field of ecology. his wide scientific interests and friendly approach to students and adepts of forest sciences were reflected in numerous works carried out by the students under his supervision. from 1980 to 2021, he supervised 301 master’s theses and 96 acta mycologica / 2021 / volume 56 / article 5611 publisher: polish botanical society 2 szczepkowski / professor andrzej piotr grzywacz dr. h. c. mult. engineering theses, and reviewed several hundred diploma theses. ese theses concerned mainly phytopathological and mycological issues as well as issues of nature protection and nature-forest education. he was a supervisor of the phytopathology section of the forestry students scientific association and forestry students scientific association of the sggw and five student scientific camps. prof. a. grzywacz is a well-liked and respected academic teacher with a rare gi of arousing interest in learning and transferring broad knowledge among students. he was the supervisor of 14 doctoral theses. his knowledge, experience, and authority led him to review 37 doctoral theses, 16 habilitation dissertations, and 17 opinions for the title of professor. for the central commission for titles and degrees, he prepared 26 opinions for professorship applications and 16 opinions for habilitation dissertations. he also prepared three reviews to grant the title of doctor honoris causa. while working as a lecturer at the warsaw university of life sciences, he served as chief nature conservator – under secretary of state in the ministry of environmental protection, natural resources and forestry, between 1992 and 1993, where he contributed to the establishment of the biebrza national park and the stołowe mountains national park, and from 1995 to 1997 he worked at the national parks board, taking care of scientists from all national parks. prof. a. grzywacz greatly values the popularization of science, on which he is working till date. his popular science books are grzyby i las [fungi and forest] (1981), grzyby leśne [forest mushrooms] (1988, 1990), grzyby chronione [protected mushrooms] (1989), poznajmy las [let’s get to know the forest] (1995), las twoim bogactwem [forest your wealth] (2000), „wiem co zbieram w lesie” [i know what i collect in the forest] (2003; together with p. staniszewski), księga lasu [e book of the forest] (2009), żywot lasu [e life of the forest] (2009), las tętniący życiem [forest teeming with life] (2010), and drzewa – pomniki przyrody [trees – nature monuments] (2013; jointly with j. pietrzak), and about 275,000 copies of these books have been published. he has published over 150 popular articles in forestry and the nature press. he coauthored three different editions of the forest protection manual (1998, 2004, 2012). he is the author of 135 mycology entries in the wielka encyklopedia powszechna pwn [great universal encyclopedia pwn] (2001–2005) and 750 entries in the słownik terminologiczny: leśnictwa, drzewnictwa, ochrony środowiska, łowiectwa i dziedzin pokrewnych [terminology dictionary: forestry, wood science, environment protection, hunting and related fields] (1996). he has presented over 280 papers in poland and abroad at conferences, symposia, and meetings of scientific societies. he has participated in over 120 radio and over 65 television programs of various types. he was a scientific consultant for nine popular science films. prof. a. grzywacz continues to hold a number of academic appointments. he is an elected member of the warsaw scientific society and the commission of agricultural, forestry and veterinary sciences of the polish academy of arts and sciences (polish academy of skills) in kraków. at the warsaw university of life sciences, he was a member of the academic senate and numerous rector’s and senate committees and an initiator and first editor-in-chief of the university quarterly agricola. at the polish academy of sciences, he used to hold the posts of chairman of the division of agricultural, forest and veterinary sciences, committee of forestry sciences, scientific councils of the botanical garden in powsin – center for the preservation of biodiversity of the polish academy of sciences. he also used to hold the position of editor-in-chief of the quarterly heureka – problems of the social scientific movement and was a member of the collegium and the presidium of the polish academy of sciences, committee on nature conservation of the polish academy of sciences, scientific councils of the institute of dendrology of the polish academy of sciences in kórnik. he continues to hold the post of vice chairman of the council of scientific societies at the polish academy of sciences. since 1979, he has been a member of the board of the polish forest society, where he also served as a vice president, and from 1997 to 2018, he was the president of the society. he is a dignified honorary member and the honorary president of the ptl. with regards to research and teaching, he was chairman of the scientific council of the intercollegiate methodological centre of agricultural academies, the scientific and didactic team for forestry and wood technology, and the forestry and wood acta mycologica / 2021 / volume 56 / article 5611 publisher: polish botanical society 3 szczepkowski / professor andrzej piotr grzywacz dr. h. c. mult. section of the scientific research committee; vice chairman of the central commission for title and scientific degrees; and member of the general council for higher education. in addition, with regards to the field of forestry and nature protection, he was the chairman of the state council for nature protection, the state forests panel, the scientific council of the forest research institute, the scientific council of the league for nature protection, the main committee of the ecological knowledge olympiad, the socio-scientific council of the lkp “puszcza knyszyńska,” and others. he continues to be the chairman or member of editorial and program boards of scientific journals: acta mycologica, folia forestalia polonica, leśne prace badawcze, phytopathologia, parki narodowe i rezerwaty przyrody, postępy nauk rolniczych, sylwan, as well as popular science magazines: las polski, poznajmy las, przyroda polska. prof. a. grzywacz has received numerous awards for his scientific and teaching activities, including awards from the minister of national education, minister of science and higher education, polish academy of sciences, minister of environment, and rector of the warsaw university of life sciences. he has been awarded the knight’s, officer’s and commander’s cross of the order of polonia restituta, the golden cross of merit, the medal of the national education commission, the michał oczapowski medal of the polish academy of sciences, medal for meritorious service to national defense, medal of the centenary of regained independence, medals of the office for veterans and repressed persons “pro patria” and “pro memoria,” medal of the polish forest society “pro bono silvae,” and badges “merited for forestry and wood industry” and “merited for environmental protection and water management.” he received great appreciation for receiving german foresters award “lorenz-wappes-preis” and the forest research institute medal named aer jan teodor hausbrandt. he also holds the cordell of the polish forester and an honorary forester’s uniform as well as the adam loret award “for many years of activity for the benefit of forestry and protection of forest nature, forest sciences and for the benefit of the state forests.” in his life, prof. grzywacz diligently follows the words of the roman philosopher seneca: “you must live for others if you want to live to benefit yourself.” although prof. a. grzywacz retired in 2013, he continues his social and popularization work very actively. he oen visits his home faculty of forestry at the warsaw university of life sciences (sggw) and willingly shares his knowledge and experience with younger employees. he also maintains the contact with colleagues from his youth, when he was actively involved in scouting, and he says that the scouts’ principles and patterns have significantly shaped his personality and influenced the choice of forestry as a way of life. prof. andrzej piotr grzywacz and his wife maria have two sons (piotr and marcin) and are happy grandparents of two granddaughters. acta mycologica / 2021 / volume 56 / article 5611 publisher: polish botanical society 4 professor andrzej piotr grzywacz dr. h. c. mult. 2014-11-20t23:21:15+0000 polish botanical society morphological and molecular characterization of entomophthorales (entomophthoromycota: entomophthoromycotina) from argentina acta mycologica article id: 5522 doi: 10.5586/am.5522 publication history received: 2020-02-27 accepted: 2020-06-26 published: 2020-11-27 handling editor malgorzata ruszkiewicz-michalska; institute for agricultural and forest environment, polish academy of sciences; university of łódź; https://orcid.org/0000-00018901-0552 authors, contributions rgm and wf conducted the dna extraction and pcr assays; abj, avt, and lac conducted the phylogenetic analyses; rgm, ccll, and lac wrote the manuscript; ccll and abj secured funding funding this study was supported by the national research council of argentina (conicet) and by university of la plata (unlp). competing interests no competing interests have been declared. copyright notice © the author(s) 2020. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. original research paper in microscopic fungi morphological and molecular characterization of entomophthorales (entomophthoromycota: entomophthoromycotina) from argentina romina g. manfrino 1*, louela a. castrillo 2, claudia c. lópez lastra 1, andrea v. toledo 3, walter ferrari 1, annette b. jensen 4 1centro de estudios parasitólogicos y de vectores – cepave (conicet, consejo nacional de investigaciones científicas y tecnológicas; unlp, universidad nacional de la plata), la plata, buenos aires, argentina 2robert w. holley center for agriculture & health, u.s. department of agriculture, agriculture research service, ithaca, 14853, ny, usa 3facultad de ciencias agrarias y forestales, centro de investigacioness de fitopatología – cidefi (cicpba, comisión de investigaciones científicas de la provincia de buenos aires; unlp, universidad nacional de la plata), la plata, buenos aires, argentina 4department of agriculture and ecology, university of copenhagen, thorvaldsensvej 40, frederiksberg c, 1871, denmark *to whom correspondence should be addressed. email: manfrino@cepave.edu.ar abstract we characterized 17 insect-pathogenic entomophthoralean fungal isolates (entomophthoromycotina: entomophthorales) using morphological and molecular techniques. we identified four species from various insect hosts: (i) entomophthora planchoniana, six specimens from aphids; (ii) pandora neoaphidis, three specimens from aphids; (iii) zoophthora phalloides from an aphid; and (iv) z. radicans, seven specimens from insects in the orders diptera, hemiptera, and lepidoptera. analysis of its1 data from e. planchoniana showed clustering in accordance to aphid host species. entomophthora planchoniana from macrosiphum euphorbiae clustered together, separate from the isolate from myzus persicae. the p. neoaphidis specimens clustered with sequences from other aphid-pathogenic pandora species in genbank. in this study, z. phalloides from brevicoryne brassicae and z. radicans from an unidentified species of chironomidae (diptera) in argentina were characterized for the first time. the present study was initiated to elucidate the taxonomy of the entomophthoralean fungi in argentina according to their morphological and molecular characters. the presented results emphasize the significance of the combination of molecular data and information on morphology, ecology, and host range for accurate identification of entomophthoralean and allied genera. keywords entomopathogens; insect pests; molecular taxonomy; host range 1. introduction entomopathogenic fungi in the subphylum entomophthoromycotina (phylum entomophthoromycota) (spatafora et al., 2016) are well-known virulent pathogens infecting many species of arthropod hosts. the subphylum contains over 300 species, most of which belong to the order entomophthorales, with nearly 290 described species (hajek et al., 2018). most of the entomophthoralean species are obligate pathogens of insects and other arthropods (gryganskyi et al., 2012; hajek et al., 2018). likewise, they are relatively host specific and, thus, pose little or no threat to non-target organisms, making these fungi ideal biological control agents against acta mycologica / 2020 / volume 55 / issue 2 / article 5522 publisher: polish botanical society 1 https://doi.org/10.5586/am.5522 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0002-2628-2259 https://orcid.org/0000-0001-8138-9764 https://orcid.org/0000-0001-6590-0171 https://orcid.org/0000-0001-6930-0077 https://orcid.org/0000-0003-3995-5122 https://orcid.org/0000-0002-2044-2274 mailto:manfrino@cepave.edu.ar manfrino et al. / morphological and molecular characterization of entomophthorales insect pests. the host specificities of different entomophthoroid genera within the entomophthorales are often limited to a given insect order or family; however, individual entomophthoroid species may show a higher level of specificity toward host genera or species (humber, 2016). however, species or strains selected for commercialization as biological control agents cannot have too narrow a specificity spectrum to be economically profitable. the identification of new strains to the species level is the first step in utilizing the full potential of fungi for specific applications (lieckfeldt et al., 1999). entomophthoralean species are identified on the basis of their host insect species and morphological features, primarily the size and shape of the primary conidia and the number of nuclei per conidium. however, in some cases, morphometric studies are insufficient to ascertain whether they represent a single taxon or complexes of morphologically similar species (barta & cagáň, 2006). for this purpose, methods based on either biochemical reactions or dna sequences are used. various molecular techniques have been used in the systematics of fungi to assess interspecific variation and determine phylogenetic relationships. up to date molecular phylogenetic studies of entomophthoromycotina have used molecular markers such as rdna (i.e., 18s, 28s, or the whole ribosomal operon) and protein coding regions (e.g., actin, β tubulin, and rpb2). in addition, other more variable regions (e.g., its) have been used to study closely related taxa (jensen et al., 2007). different combinations of these have been used in multiple gene analysis (gryganskyi et al., 2013). today, the vast majority of available genomic information within the entomophthoromycotina is composed of partial gene and intron sequences developed for use in phylogenetic analyses. most sequences deposited in the national center for biotechnology information (ncbi) genbank database are from the nuclear ribosomal dna region, including the large (lsu) and small (ssu) subunits, 5.8s, and internal transcribed spacer (its) regions 1 and 2. only a few sequences of entomophthoralean fungi from argentina are available in genbank (jensen et al., 2009; scorsetti et al., 2011), reflecting the paucity of information on the diversity of this fungal group in the region. the present paper reports on the identification of entomophthora planchoniana cornu, pandora neoaphidis (remaud. & hennebert) humber, zoophthora phalloides a. batko, and zoophthora radicans (bref.) a. batko from argentinian insects and provides both morphological and molecular characterizations of these species. the fungal pathogens z. phalloides and z. radicans from both brevicoryne brassicae l. and from an unidentified species of chironomidae (diptera), respectively, are recorded for first time in argentina. 2. material and methods 2.1. specimens and morphological characterization specimens were collected during sampling from crops of economic importance in 2007–2013 (table 1). the isolates examined in this study, their host taxon, plant host, collection site, date, and genbank accession numbers are listed in table 1. zoophthora isolates were cultivated on sabouraud dextrose agar plus 1% yeast extract (sday 1%), a suitable medium used for the isolation and culture of several species of entomophthorales (choi et al., 2016; feng et al., 1990; moubasher et al., 2010; zhou et al., 2016). pandora and entomophthora specimens were identified directly from infected insect hosts. specimens were examined under a stereomicroscope and an optical microscope for the presence of rhizoids, cystidia, and/or conidia. fungal structures were mounted in lactophenol-aceto-orcein (lpao) (1:1) or stained with 1% aceto-orcein plus glycerine for semipermanent mounts (as preserved material for fungarium) and measured. fungal species were identified according to taxonomic monographs and the keys of bałazy (1993), keller (2007), and humber (2012). photographs of the primary and capilliconidia of z. phalloides and z. radicans from in vitro cultures were taken with an olympus bx51 camera (japan) at ×400 magnification. aphid host identification to the species level acta mycologica / 2020 / volume 55 / issue 2 / article 5522 publisher: polish botanical society 2 manfrino et al. / morphological and molecular characterization of entomophthorales was made using the keys of blackman and eastop (2000). lepidopteran hosts were identified by a taxonomic specialist at the instituto nacional de tecnología agropecuaria (inta) (see acknowledgments). the specimens were preserved in the mycological collections at the centro de estudios parasitológicos y de vectores (cepave, la plata, argentina). the isolates were deposited in the mycological collections at cepave and the usda-ars collection of entomopathogenic fungal cultures (arsef) in ithaca, new york. 2.2. molecular characterization: dna extraction and pcr amplification fungus-infected insect cadavers were stored in 96% ethanol, and in vitro cultures were prepared as described by jensen et al. (2001) until dna extraction. genomic dna was extracted from infected insects (in vivo; for entomophthora and pandora specimens) or from in vitro cultures (for zoophthora spp. specimens) via chelex extraction (traugott et al., 2008) or by using a dneasy plant mini kit (qiagen, md, usa) following the manufacturer’s protocol. pcr assays were performed on three loci, the internal transcribed spacer 1 region (its1), the 18s gene (ssu), and the first part of the 28s gene (lsu). because of the low quantity of dna obtained from infected insects, pcr assays of entomophthora and pandora specimens were limited to the its1 and lsu only. universal fungal primers or entomophthoraleanspecific primers were used to avoid amplification of insect host dna. we used the universal fungal primers its5 (white et al., 1990) and the single reverse primer nu5.8s-3′ (jensen & eilenberg, 2001) the universal fungal primers nu-ssu-0021-58 (gargas & depriest, 1996) and nu-ssu-1780-38 (depriest, 1993) for the ssu rdna region, and nu-lsu-0018-5 (jensen & eilenberg, 2001) and lsu-0805 (kjøller & rosendahl, 2000) for lsu amplification. the pcr conditions were: initial denaturation for 30 seconds at 98 ◦c, followed by 38 cycles of denaturation for 10 seconds at 98 ◦c, annealing for 20 seconds at 55–60 ◦c (its1 60 ◦c, ssu 58 ◦c, lsu 55 ◦c), extension for 1 min at 72 ◦c, and a final extension for 10 min at 72 ◦c. the pcr reactions were carried out in 50 µl volumes, with 250 µm of each dntp, 0.8 µm of each primer, 2.5 mm mgcl2, 1× buffer (10 mm tris-hcl, ph 8.8 at 25 ◦c, 50 mm kcl, 0.1% triton x100), 1 unit of dynazyme ii (finnzymes, espoo, finland), and 1 µl of extracted dna (diluted 1:10). the size of the pcr amplification products was estimated by electrophoresis on a 1.5% agarose gel in 0.5× tbe, and the products were visualized with ez-vision (amresco, usa). pcr products were purified using the qiaquick purification kit (qiagen, md, usa) following the manufacturer’s protocol. amplicon sizes were checked by electrophoresis, and purified pcr products were sent to macrogen (seoul, korea) for sequencing in both directions. the sequences obtained were submitted to ncbi genbank (https://www.ncbi.nlm.nih.gov/) for gene annotation. sequences were edited using bioedit version 7.0.9.0 (hall, 1999) and were used to perform a phylogenetic analysis that included some sequences of related species in the genera entomophthora, pandora, and zoophthora available at genbank. sequence data for each locus were aligned with the clustalw tool of mega5 (tamura et al., 2011) and trimmed, as needed, using mesquite version 3.0.2 (maddison & maddison, 2009). aligned and trimmed sequence data were submitted to treebase (http://purl.org/phylo/treebase/phylows/study/tb2:s27276?xaccess-code=5af44733057f582185683187bae29cbf&format=html). each dataset was analyzed using maximum parsimony (mp) in paup version 4.0a142 (swofford, 2002) to determine the number of mp-informative sites. the phylogenetic analysis was carried out using the maximum likelihood (ml) method. the consensus tree was built using the latest 1,000 trees. statistical support for the nodes was evaluated using 1,000 replicates. phylogenetic analysis was limited to each locus; incomplete its1, lsu, and ssu sequence data from all specimens prevented presentation of a multilocus tree. acta mycologica / 2020 / volume 55 / issue 2 / article 5522 publisher: polish botanical society 3 m an frin o et al./ m o rp h o lo g icalan d m o lecu larc h aracterizatio n o fen to m o p h th o rales table 1 list of species used in the study with information about insect and plant host taxa, location of collection, and genbank accession numbers. species/strain number (cep) or exsiccate number (cephe) insect host plant host location genbank accession numbers its1 ssu lsu town/city geographical coordinates latitude s longitude w entomophthora planchoniana cephe61 myzus persicae (sulzer) capsicum annuum l. monte vera 31◦32′58.21′′ 60◦41′34.74′′ mg252293 cephe62 macrosiphum sp. rosa sp. sunchales 30◦56′00′′ 61◦34′00′′ mg252624 cephe65 myzus persicae (sulzer) solanum melongena l. monte vera 31◦32′58.21′′ 60◦41′34.74′′ mg256477 cephe63 macrosiphum sp. rosa sp. sunchales 30◦56′00′′ 61◦34′00′′ mg256478 cephe64 macrosiphum sp. rosa sp. sunchales 30◦56′00′′ 61◦34′00′′ mg256479 mh366738 cephe66 myzus persicae (sulzer) capsicum annuum l. monte vera 31◦32′58.21′′ 60◦41′34.74′′ mg256476 pandora neoaphidis cephe72 myzus persicae (sulzer) solanum melongena l. monte vera 31◦32′58.21′′ 60◦41′34.74′′ mh366634 cephe73 nasonovia ribisnigri (mosley) lactuca sativa l. monte vera 31◦32′58.21′′ 60◦41′34.74′′ mh366798 mh366734 cephe74 rhopalosiphum padi l. avena sativa l. rafaela 31◦12′6.62′′ 61◦30′11.14′′ mh366739 zoophthora phalloides cep 687 (arsef 11861) brevicoryne brassicae l. brassica oleracea var. botrytis l. monte vera 31◦32′58.21′′ 60◦41′34.74′′ mg253005 z. radicans cephe67 plutella xylostella l. brassica napus l. ataliva 30◦59′00′′ 61◦27′00′′ mg256481 mg252955 mg256485 cephe68 acyrthosiphon pisum (harris) medicago sativa l. monte vera 31◦32′58.21′′ 60◦41′34.,74′′ mg256480 mg252969 mg256487 cephe69 brevicoryne brassicae l. brassica oleracea var. botrytis l. monte vera 31◦32′58.21′′ 60◦41′34.74′′ mg256482 mg252968 mg256488 cephe70 plutella xylostella l. brassica napus l. rafaela 31◦12′6.62′′ 61◦30′11.14′′ mg256486 mg256489 cephe71 plutella xylostella l. brassica oleracea var. capitata l. monte vera 31◦32′58.21′′ 60◦41′34.74′′ mg256483 mg252997 mg256490 cep 30 (arsef 6917) tuta absoluta (meyrick) solanum lycopersicum l. colonia urquiza 34◦56′19.2′′ 58◦06′3.8′′ mg256484 mg256492 cep 320 (arsef 8466) chironomidae unknown la plata 34◦56′00′′ 57◦57′00′′ mg256491 a cta m ycologica / 2020 / vo lu m e 55 / issu e 2 / a rticle 5522 pu b lish er:po lish b o tan icalso ciety 4 manfrino et al. / morphological and molecular characterization of entomophthorales 3. results 3.1. fungal identification and taxonomic observations based on morphological characters, four fungal species were identified in the 17 insect specimens: e. planchoniana, p. neoaphidis, z. phalloides, and z. radicans (table 1). 3.1.1. entomophthora planchoniana cornu bull. soc. bot. fr. 20: 189 (1873). hosts. myzus persicae sulzer and macrosiphum sp. (hemiptera: aphididae). description. primary conidia campanulate, 14.2 ± 2.5 (12.1–16.8) × 12.1 ± 2.3 (9.8– 14.5) µm, with a small apiculus and broad, nearly flattened papilla, and multinuclear (three–five nuclei). secondary conidia were observed emerging from the primary conidia and were slightly smaller: 11.2 ± 1.72 (9.7–13.5) × 10.5 ± 1.5 (8.7–12.2) µm (figure 1e). numerous thin rhizoids (figure 1f). cystidia and resting spores not observed. attempts to obtain fungal isolates were not successful. fungarium accession numbers. cephe61, cephe66 (m. persicae from capsicum annuum l.), cephe62, cephe63, cephe64 (macrosiphum sp. from rosa sp.), and cephe65 (m. persicae from solanum melongena l.). 3.1.2. pandora neoaphidis (remaud. & hennebert) humber mycotaxon 34 (2): 452 (1989). hosts. nasonovia ribisnigri (mosley), m. persicae, and rhopalosiphum padi l. (hemiptera: aphididae). description. primary conidia ovoid or elongated, 23.55 ± 2.07 (19.6–27.3) × 12.08 ± 1.55 (9.6–14.3) µm, generally conical papilla connected gently with the body of the conidia secondary conidia spherical or bell-shaped, 17.33 ± 1.36 (15.2–20.1) × 13.37 ± 1.26 (11.16–15.32) µm (figure 1g). capilliconidia absent. rhizoids with discoid or irregularly branched adhesive disc. cystidia present. no resting spores were observed. attempts to obtain fungal isolates were not successful. fungarium accession numbers. cephe72 (m. persicae from s. melongena), cephe73 (n. ribisnigri from lactuca sativa l.), and cephe74 (r. padi from avena sativa l.). 3.1.3. zoophthora phalloides a. batko acta mycologica 2: 8 (1966). host. brevicoryne brassicae (hemiptera: aphididae). description. primary conidia elongated cylindrical to elongated oval, 33.03 ± 4.89 (24.08–42.16) × 12.3 ± 2.52 (9.92–17.36) µm (figure 1c). this characterization was in accordance with the original description (batko, 1966). the presently observed secondary conidia were capilliconidia: 18.40 ± 2.67 (12.4– 22.32) × 7.04 ± 1.22 (4.96–9.92) × 94.63 ± 9.98 (79.36–114.08) µm (figure 1d). rhizoids were ramified and forming rhizomorphs. no resting spores were observed. culture collection accession number. cep 687 (arsef 11861) (b. brassicae from brassica oleracea var. botrytis l.). 3.1.4. zoophthora radicans (bref.) a. batko bull. acad. polon. sci., cl. ii. sér. sci. biol. 12: 323 (1964). hosts. acyrthosiphon pisum harris, b. brassicae (hemiptera: aphididae), plutella xylostella l. (lepidoptera: plutellidae), tuta absoluta (meyrick) (lepidoptera: gelechiidae), and an unidentified species of chironomidae (diptera). acta mycologica / 2020 / volume 55 / issue 2 / article 5522 publisher: polish botanical society 5 manfrino et al. / morphological and molecular characterization of entomophthorales description. elongated primary conidia uninucleate, 22.63 ± 3.40 (16.36–25.94) × 8.14 ± 0.75 (7.39–9.67), generally conical papilla demarcated with a slight protuberance from the body of the conidia (figure 1a). secondary conidia similar to primary ones or capilliconidia: 20.04 ± 1.83 (18.89–24.4) × 7.09 ± 0.54 (6.39– 7.93) × 46.95 ± 9.70 (35.81–60.31) µm, fusiform and formed laterally on slender, capillary conidiophores arising from primary conidia (figure 1b). rhizoids mostly in pseudorhizomorphs, particularly abundant in the thoracical part, single or fasciculate with specialized adhesive discs. conidiophores branched with terminal enlargement. cystidia 3.69 µm in diameter, tapering uniformly at the base. no resting spores were observed. fungarium accession numbers. cephe67 (p. xylostella from brassica napus l.), cephe68 (a. pisum from medicago sativa l.), cephe69 (b. brassicae from b. oleracea var. botrytis), cephe70 (p. xylostella from b. napus), and cephe71 (p. xylostella from brassica oleracea var. capitate l.). culture collection accession numbers. cep 30 (arsef 6917; t. absoluta from solanum lycopersicum l.) and cep 320 (arsef 8466; indet. diptera from indet. host plant). figure 1 zoophthora radicans from tuta absoluta (meyrick) primary conidium (a) and two capilliconidia each on a capilliconidiophore produced from a primary conidium (a,b). bar: 10 µm. zoophthora phalloides primary conidia (c) and capilliconidia on capilliconidiophore from primary conidium (d). bar: 10 µm. entomophthora planchoniana from myzus persicae (sulzer) primary and secondary conidia (e). bar: 20 µm. rhizoids of entomophthora planchoniana from myzus persicae (sulzer) (f). bar: 1 mm. pandora neoaphidis from nasonovia ribisnigri (mosley) primary and secondary conidia (g). bar: 10 µm. 3.2. molecular characterization: dna sequencing and molecular analyses we successfully amplified dna samples and obtained sequences of the its1 (337 bp for z. radicans, 259 bp for e. planchoniana, and 308 bp for p. neoaphidis), lsu (755 bp for z. radicans, 855 bp for p. neoaphidis, and 820 bp for e. planchoniana), and ssu (1,720 bp for z. radicans and 1,739 bp for z. phalloides). all sequences were deposited in genbank under the accession numbers given in table 1. combined with the genbank sequences of other entomophthoralean fungi, paup analysis showed: its1 sequence data (total 733 bp) with 141 variable parsimonyuninformative and 541 parsimony-informative characters, lsu sequence data acta mycologica / 2020 / volume 55 / issue 2 / article 5522 publisher: polish botanical society 6 manfrino et al. / morphological and molecular characterization of entomophthorales (total 1,024 bp) with 165 variable parsimony-uninformative and 341 parsimonyinformative characters, and ssu sequence data (total 1,863 bp) with 207 variable parsimony-uninformative and 54 parsimony-informative characters. the best tree from the ml analysis of the its1 sequences showed that our isolates of e. planchoniana and p. neoaphidis clustered with the same respective entomophthoralean species, with a 67% bootstrap value (figure 2). all e. planchoniana specimens had the same its1 sequence. the its1 sequence of p. neoaphidis cephe73 clustered with p. neoaphidis arsef 835 in another monophyletic group supported by a 99% bootstrap value. finally, the z. radicans cephe67, cephe68, cephe69, cephe70, cephe71, and cep 30 sequences clustered with z. radicans nw386 in another monophyletic group supported by a 100% bootstrap value. the its sequences of the z. radicans specimens did not reveal differences between different hosts. figure 2 phylogenetic relationships of zoophthora, pandora, and entomophthora and related species inferred from maximum likelihood analysis of its1 sequences. the sequences corresponding to this work are marked with asterisks. bootstrap values are noted above the internodes. the bar at the bottom indicates the number of substitutions per site. in the phylogenetic analyses of the lsu rrna, p. neoaphidis and z. radicans were clustered in a monophyletic group with sequences from related entomophthoralean species. this grouping was supported by a 100% bootstrap value. the lsu sequences of six z. radicans specimens from diptera, hemiptera, and lepidoptera were identical to that of arsef 388, which was isolated from a dipteran in switzerland. zoophthora radicans specimen 320 had 99% similarity with the group (figure 3). entomophthora planchoniana specimen cephe64 clustered with e. planchoniana arsef 6252 in another monophyletic group, which was supported by a 100% bootstrap value (figure 3). in the ml tree from the phylogenetic analysis of the ssu rrna sequences, z. radicans cephe67, cephe68, cephe69, and cephe71 were clustered in a monophyletic group supported by a 65% bootstrap value, which includes representatives of z. radicans (z. radicans arsef 853) and z. lanceolata (z. lanceolata arsef 469) (figure 4). based on the ssu rrna sequences, z. phalloides cep 687 clustered with representatives of z. anglica, z. phalloides, and z. occidentalis in another monophyletic group supported by a 99% bootstrap value. likewise, the sequence of z. phalloides cep 687 characterized in this work was clustered with z. phalloides arsef 2281 in a same clade supported by a 100% bootstrap value (figure 4). 4. discussion in the present study, the initial identification of fungi from various insect hosts was based mainly on the shape and size of the primary and secondary conidia and acta mycologica / 2020 / volume 55 / issue 2 / article 5522 publisher: polish botanical society 7 manfrino et al. / morphological and molecular characterization of entomophthorales figure 3 phylogenetic relationships of zoophthora, pandora, and entomophthora and related species inferred from maximum likelihood analysis of lsu rdna sequences. the sequences corresponding to this work are marked with asterisks. bootstrap values are noted above the internodes. the bar at the bottom indicates the number of substitutions per site. figure 4 phylogenetic relationships of zoophthora and related species inferred from maximum likelihood analysis of ssu rdna sequences. the sequences corresponding to this work are marked with asterisks. bootstrap values are noted above the internodes. the bar at the bottom indicates the number of substitutions per site. according to host affinity. these identifications were confirmed by use of molecular data, specifically the dna sequences of the its1, lsu, and ssu. based on the results obtained in this study, the morphology of the secondary conidia of p. neoaphidis was consistent with the original descriptions reported by humber (1989). our results were also consistent with observations made by scorsetti et al. (2006). these authors previously reported p. neoaphidis from aphids in argentina, with measurements of both primary and secondary conidia very similar to those observed in this study. with regards to its insect hosts, p. neoaphidis is a common aphid pathogen with a cosmopolitan distribution. in argentina, it was previously recorded in 17 aphid species (lópez lastra & scorsetti, 2006; lópez lastra et al., 2019; manfrino, hatting, et al., 2014; manfrino et al., 2013; scorsetti et al., 2006) and was reported by scorsetti et al. (2006) as the most predominant pathogen of aphids in argentina. we identified six specimens of entomophthora from aphids as e. planchoniana. the identity of the fungal species was determined based on insect host, number/size of conidial nuclei, and conidial size (humber, 2012). compared to the original acta mycologica / 2020 / volume 55 / issue 2 / article 5522 publisher: polish botanical society 8 manfrino et al. / morphological and molecular characterization of entomophthorales description of e. planchoniana (keller, 1991), the materials observed in this study had slightly smaller conidia than those originally described. the results obtained in this study, however, were consistent with those made by lópez lastra and scorsetti (2006) and scorsetti et al. (2006). these authors previously recorded e. planchoniana as a pathogen of n. ribisnigri, aphis fabae, aphis gossypii, myzus sp., and m. persicae in argentina and observed that the primary conidia from their specimens were slightly smaller than those described by keller (1991). for many years, e. chromaphidis was considered to be a synonym for e. planchoniana (gustafsson, 1965; macleod et al., 1976; waterhouse & brady, 1982) until these two species were definitely separated and justified (humber & feng, 1991). entomophthora chromaphidis occurs in north america and australia and is characterized as having smaller primary conidia and larger nuclei, whereas e. planchoniana is more widely distributed (primarily in europe) and has larger primary conidia and smaller nuclei. in argentina, e. planchoniana has been recorded as an aphid pathogen on horticultural crops (lópez lastra & scorsetti, 2006; lópez lastra et al., 2019; scorsetti et al., 2006). it has also been recorded causing epizootics in m. persicae on pepper and eggplant crops (manfrino et al., 2016). regarding its host affinity, previous studies have already covered entomophthora species. a study elucidated the host-driven divergence of species within the e. muscae (cohn) fresen. species complex and e. planchoniana cornu (jensen et al., 2009). thomsen and jensen (2002) showed a separation of resting spore isolates of e. muscae species complex at the species level, which is not possible using only morphological characters (i.e., diameter). freimoser et al. (2001) and jensen et al. (2009) showed that isolates originating from different specimens of the same host taxa appeared to be strongly clonal, even when they were sampled at different localities in different years. in agreement with these observations, our study showed only minor differences in e. planchoniana specimens from several different aphid species based on its1 sequences. in this study, the primary and secondary conidia of zoophthora radicans specimens were slightly bigger than the original description reported by keller (1991). our results are consistent with reports by lópez lastra and scorsetti (2006) regarding the measurements of the primary conidia of z. radicans from t. absoluta, although we observed bigger capilliconidia. scorsetti et al. (2006) recorded z. radicans from six aphid species, and our measurements of the primary and secondary conidia of this species are consistent with their data. additionally, these authors recorded zoophthora sp. from only one aphid species and found that its morphological characteristics clearly differed from those of z. radicans. in this study, measurements of zoophthora from b. brassicae were different from those of z. radicans, with primary and secondary conidia of greater values. based on the original description by batko (1966), this species corresponds to z. phalloides. glare et al. (1987) studied different criteria based on morphological, physiological, and biochemical characters in order to reliably differentiate z. phalloides from z. radicans. this allowed the division of isolates broadly defined as z. radicans and z. phalloides using shape and spore dimensions, host specificity, growth in vitro, and analyses of isoenzymes and fatty acid composition (glare et al., 1987). balazy (1993) conducted thorough studies on the morphology, biology, and particularly host specificity of the genus zoophthora, which enabled him to distinguish and describe seven new species morphologically similar to z. radicans. zoophthora phalloides is very similar morphologically to other aphid pathogenic members of the zoophthora spp. complex. zoophthora phalloides was first described as a pathogen of aphids (hemiptera) in poland by batko (1966) and has since been recorded in north america, britain, denmark, france, switzerland, israel, korea, new zealand, and occasionally in australia (e.g., barta & cagáň, 2005; nielsen et al., 2001; papierok et al., 2016; yoon et al., 1998), thus indicating a worldwide distribution. zoophthora radicans has the broadest host range reported for any species of the entomophthoraceae. this observation has led some authors to suggest that it is a complex of several species differing in their pathogenicity to different hosts (bałazy, 1986; glare, 1988; humber, 1983; mcguire et al., 1987). it has acta mycologica / 2020 / volume 55 / issue 2 / article 5522 publisher: polish botanical society 9 manfrino et al. / morphological and molecular characterization of entomophthorales been collected worldwide from hosts in the insect orders lepidoptera, diptera, coleoptera, orthoptera, thysanoptera, hemiptera, and hymenoptera (glare & milner, 1991; keller, 1991; milner & soper, 1981; papierok et al., 1984; soper & ward, 1981). in argentina, z. radicans has been reported as a pathogen of several insect species (manfrino, hatting, et al., 2014; manfrino et al., 2013; manfrino, zumoffen, et al., 2014; scorsetti et al., 2006), but there were no records of dipteran hosts until this study. milner and mahon (1985) identified z. radicans in other species of diptera (damaromyia sp., sciaridae, psychodidae) in australia. our results revealed that the its1, lsu, and ssu sequences of z. radicans specimens from diptera, lepidoptera, and hemiptera show few differences. in analyses of lsu sequences, all specimens clustered as z. radicans, with a slight difference for the dipteran isolate, in agreement with reports of its wide host range. for species included within a species complex, molecular characters are critical data supplementing analysis at different taxa. in this study, species of zoophthora in argentina were characterized by molecular tools for the first time. molecular analyses based on ssu rdna sequences confirmed the identity of z. phalloides, with the argentinean strain grouped with z. phalloides strain arsef 2281 (gb ef392558) with a 100% bootstrap value. likewise, with regards to host affinity, the zoophthora aphid-pathogenic isolate cep 687 was clustered with isolate arsef 2281 (z. phalloides), which was originally isolated from b. brassicae, in accordance with the host aphid species of our native strain (figure 4). this study reports the first record of z. phalloides as a pathogen of b. brassicae and z. radicans as a pathogen of an unidentified species of chironomidae (diptera) in argentina. the identification and establishment of in vitro cultures of these fungal species would allow further studies determining their potential as microbial biological control agents for the management of insect pests of agricultural crops. of the four fungi, zoophthora spp. and pandora neoaphidis can be grown on sday, but zoophthora spp. are easier to cultivate than p. neoaphidis on this medium. other entomophthoralean species included in the genus entomophthora require complex and highly nutritious culture media, which is considered a sign of high specialization and close adaptation to the host (humber, 1994). based on these observations, zoophthora strains would have greater potential to be used as targeted microbial control agents. acknowledgments we thank michael griggs of the usda-ars (eppru, ithaca, ny) for technical assistance and césar salto, phd for the identification of lepidopteran host species. we also thank the reviewers for their helpful comments. references bałazy, s. 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(2016). a floatable formulation and laboratory bioassay of pandora delphacis (entomophthoromycota: entomophthorales) for the control of rice pest nilaparvata lugens stål (hemiptera: delphacidae). pest management science, 72, 150–154. https://doi.org/10.1002/ps.3981 acta mycologica / 2020 / volume 55 / issue 2 / article 5522 publisher: polish botanical society 13 https://doi.org/10.1007/bf02372215 https://doi.org/10.1007/s10526-006-9045-1 https://doi.org/10.1016/j.funbio.2011.11.002 https://doi.org/10.3852/16-042 https://doi.org/10.1093/molbev/msr121 https://doi.org/10.1080/15572536.2003.11833173 https://doi.org/10.1111/j.1365-294x.2008.03878.x https://doi.org/10.1016/s0007-1528(82)80006-0 https://doi.org/10.1016/s0007-1528(82)80006-0 https://doi.org/10.1002/ps.3981 introduction material and methods specimens and morphological characterization molecular characterization: dna extraction and pcr amplification results fungal identification and taxonomic observations entomophthora planchoniana cornu pandora neoaphidis (remaud. & hennebert) humber zoophthora phalloides a. batko zoophthora radicans (bref.) a. batko molecular characterization: dna sequencing and molecular analyses discussion acknowledgement references professor maria rudawska acta mycologica article id: 5524 doi: 10.5586/am.5524 publication history received: 2020-07-01 accepted: 2020-07-18 published: 2021-02-15 handling editor wojciech pusz; wrocław university of environmental and life sciences, poland; https://orcid.org/0000-00031531-2739 funding the work was supported by the institute of dendrology, polish academy of sciences. competing interests no competing interests have been declared. copyright notice © the author(s) 2020. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. memories and scientists professor maria rudawska tomasz leski * institute of dendrology, polish academy of sciences, parkowa 5, kórnik, 62-035, poland *to whom correspondence should be addressed. email: tleski@man.poznan.pl maria rudawska was born on june 17, 1949, in ostrów wielkopolski, where she lived until she completed her primary and secondary education. in 1967, she began studying biology at the faculty of biology and earth sciences at the adam mickiewicz university in poznań. in 1971, she received a scholarship funded by the department of dendrology (now institute of dendrology), polish academy of sciences, in kórnik. in 1972, based on her work, nitrogen fractions and reducing sugars in isolated lupine cotyledons written under the supervision of dr. aleksandra hoffmann from the department of plant physiology, prof. rudawska received a master’s degree in biology with an emphasis in botany. she started working at the institute of dendrology immediately after graduating, and kórnik became her new home. she has spent the entire path of her scientific career at the institute of dendrology; her scientific experience in mycological studies comes from nearly five decades of work at the institute. in 1974, she became an assistant at the laboratory of tree physiology headed by professor mirosław tomaszewski. at that time, prof. rudawska’s scientific relationships included prof. roman pachlewski, from the forest research institute, who introduced her to the fascinating world of mycorrhizal fungi, after which she enthusiastically took up research on the physiology of forest tree mycorrhiza (mainly scots pine). she also worked on a research topic headed by prof. tomaszewski, “poplar resistance to infection caused by the fungus dothichiza populea,” financed acta mycologica / 2020 / volume 55 / issue 2 / article 5524 publisher: polish botanical society 1 https://doi.org/10.5586/am.5524 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0003-4167-5015 mailto:tleski@man.poznan.pl leski / professor maria rudawska by the polish-american marie skłodowska-curie foundation. in subsequent years, she began research on the synthesis of auxins and cytokinins by various mycorrhizal fungal symbionts of scots pine in relation to nitrogen and phosphorus nutrition. this issue was the basis of her doctoral dissertation titled the effect of mineral nutrition on the auxins and cytokinins production in pine mycorrhiza prepared under the supervision of prof. tomaszewski. the defence of her doctoral dissertation took place in december 1981 at the faculty of biology and earth sciences at the adam mickiewicz university in poznań during the first days of martial law. professor rudawska remembers her phd thesis defence as a very abnormal situation, riddled with fear and anxiety. in 1983, during her 4-month internship, prof. rudawska stayed at the institute of plant protection in merelbeke, belgium, where she became acquainted with the biochemical mechanisms of the antagonistic properties of the fungi trichoderma viride and t. harzianum in relation to certain pathogens. after returning, she continued this research along with studies on mycorrhiza and mycorrhizal fungi. in the following years of her scientific career, prof. rudawska continued to explore factors regulating the establishment of mycorrhizal symbiosis. some of her research in this field was carried out during her two half-year internships, 1987–1988 and 1993–1994, at claude bernard university in lyon, france, in cooperation with dr. gilles gay and dr. jean-claude debaud. the research conducted in france examined the role of auxins and enzymes of nitrogen metabolism in the process of mycorrhiza formation, including ectomycorrhizal and ectendomycorrhizal fungi. in some studies, she used a genetically transformed strain of the hebeloma cylindrosporum with auxin overproduction as a model organism. professor rudawska summarized the results of these studies in her postdoctoral dissertation titled, the key factors affecting mycorrhizal symbiosis of scots pine seedlings, defended in 1998, at nicolaus copernicus university in toruń. nothing in nature is immutable, and prof. rudawska’s scientific interests continued to evolve. after an intensive period of physiological research on ectomycorrhiza, she switched her attention to the problem of forest decline and the role of mycorrhizae and mycorrhizal fungi in this process. her work in this area included laboratory and field studies. laboratory experiments focused on the effects of low ph and increased aluminum ion content on the growth of ectomycorrhizal fungi and the functioning of mycorrhizal symbiosis. in the field studies, she analyzed the impact of environmental pollution from different sources, in particular, copper and aluminum smelters, phosphate fertilizer plants, and the urban-industrial area of kraków, on the diversity and species composition of communities of ectomycorrhizal fungi associated with forest trees. professor rudawska’s participation in the 1998 second international conference on mycorrhizae, held in uppsala, sweden, inspired the introduction of molecular methods to identify mycorrhizal fungi from environmental samples. with her research team, professor rudawska was one of the first mycologists in poland to introduce such analyses as a standard technique. over time, it turned out to be a right step; molecular analysis techniques have greatly advanced the identification of fungal species, even from a single ectomycorrhizal root tip. identification of ectomycorrhizas based on the sequencing of fungal its rdna allowed prof. rudawska’s team to develop intensive research on ectomycorrhizal fungal communities accompanying forest trees in different environmental conditions. her studies have involved, among other topics, chronosequences of fungi accompanying scots pine and european larch, the influence of tree genotype and environment on mycorrhizal communities of scots pine and poplars, and the diversity of fungi associated with alien tree species. in recent years, prof. rudawska’s research has focused on studies of fungal communities in protected and managed mixed coniferous forests. professor rudawaska’s research has determined that the species composition of fungal communities is largely similar between forest reserves and managed forests, and only a small pool of species are unique to one of the stand types. such results suggest that forest reserves as well as managed forests contribute to maintaining fungal diversity. acta mycologica / 2020 / volume 55 / issue 2 / article 5524 publisher: polish botanical society 2 leski / professor maria rudawska for several years, prof. rudawska has taken particular interest in ectomycorrhizal fungi associated with tree seedlings. she has conducted several studies on the diversity of ectomycorrhizal fungi in bare-root forest nurseries, and her results have been published as a series of 13 scientific papers. her research clearly evinces that seedlings of forest tree species such as scot pine, norway spruce, european larch, oaks, european beech, silver birch, european hornbeam, and small-leaved lime produced in bare-root forest nurseries are well colonized by ectomycorrhizal fungi, and these fungal communities are characterized by high species diversity. over many years of research, prof. rudawska’s efforts have contributed to the identification of more than 80 species of fungi in polish forest nurseries. during her many years of work at the institute of dendrology, prof. rudawska has played various roles: she was head of the institute’s physiology department and head of the laboratory of mycorrhizal research. currently, she heads the laboratory of symbiotic associations. she has also served as deputy chairman of the scientific council of the institute of dendrology for several terms. professor rudawska will be remembered at the institute of dendrology not only for her research, but also as a generous team player; she founded a mycorrhizal group with talented contributors that now extends through multiple generations of researchers, all still contributing to her fondest interests – mycology, symbiosis and plant physiology, and biochemistry. while working in kórnik, prof. rudawska participated in numerous scientific internships abroad at a wide array of institutions, including the institute of experimental botany of the czechoslovak academy of sciences, praha, czechoslovakia; institute of landscape ecology, pruhonice, czechoslovakia; finnish forest research institute, helsinki, finland; swedish university of agricultural sciences department of forest mycology and pathology, uppsala, sweden; inraflore pathogene, dijon, france; and slovenian forestry institute, ljubljana, slovenia. professor rudawska has been tremendously successful in securing funds for scientific research. she has managed many research projects financed by the foundation for polish science, the national fund for environmental protection and water management, the state committee for scientific research, the ministry of science, the national science center, as well as the general directorate of the state forests. professor rudawska’s numerous academic achievements include more than 80 original scientific papers, published mostly in recognized international peerreviewed scientific journals such as new phytologist, mycorrhiza, forest ecology and management, fungal ecology, annals of forest sciences, physiologia plantarum, chemosphere, environmental pollution, applied and environmental microbiology, and microbial ecology and indexed by clarivate analytics (web of science) and scopus. a vital aspect of prof. rudawska’s scientific activity is her long-standing cooperation with dr. algis aučina from the botanical garden of vilnius university, lithuania, resulting in many scientific papers. prof. rudawska is also the author of numerous popular science publications and chapters in monographs, including chapters on mycorrhiza of scots pine, norway spruce, european beech, oaks, and elms published by the institute of dendrology in forest tree monographs. during her scientific career, she has actively participated in more than 50 international conferences, including all the european symposia on mycorrhizae, most international conferences on mycorrhizae, and three congresses of european mycologists. professor rudawska was also an active member of several cost actions focused on fungal belowground biodiversity. professor rudawska has supervised four doctoral theses, nine master’s theses, and three bachelor’s theses. she has been a reviewer for more than 30 doctoral theses, habilitation dissertations, and professorship proceedings and more than 80 manuscripts submitted to scientific journals. owing to her scientific acumen and experience, professor rudawska is often invited to review scientific projects for various domestic and foreign entities. for many years, she has worked as an independent expert and rapporteur at the european commission (fp7, h2020). although prof. rudawska’s work at the scientific institute does not involve direct teaching responsibilities, she has always been generous with her time for students acta mycologica / 2020 / volume 55 / issue 2 / article 5524 publisher: polish botanical society 3 leski / professor maria rudawska and repeatedly presented guest lectures and laboratory classes for students of forestry and biology visiting the institute of dendrology. as a result of her significant scientific achievements and contributions, prof. rudawska was awarded the title of full professor in biological sciences by the president of the republic of poland, lech kaczyński, in a ceremony held at the presidential palace on june 8, 2006. professor rudawska is a member of many scientific societies including the polish mycological society (founding member), the polish forestry society, the european mycological association, and the federation of european society of plant physiologists. she is also an active member of the mycological section of the polish botanical society where she served as the secretary of the poznań division of the section for several years, and as chairperson of the mycology section from 2010 to 2016. from 2014 to 2018, she was also the editor-in-chief of the journal acta mycologica. at that time, acta mycologica became one of the publications indexed by the scopus database. she is also a member of the national committee on forestry sciences and wood technology of the polish academy of sciences and the committee on forestry sciences and wood technology of the polish academy of sciences – poznań branch. in her professional life, prof. rudawska has constantly been guided by one basic principle: regardless of the situation, you should be a decent person, and always try to do the best job you can. in her personal life, prof. rudawska is a very active traveler and a great lover of the baltic sea and of flowers; her immense passion for classical music led her to bestow her dog with the mellifluous name bemol. she is a mother to three sons, and a happy grandmother to five grandchildren – two girls and three boys. acta mycologica / 2020 / volume 55 / issue 2 / article 5524 publisher: polish botanical society 4 addendum to the mycobiota of smut fungi in poland 1 of 5published by polish botanical society acta mycologica short communication addendum to the mycobiota of smut fungi in poland agata wołczańska1*, anna cwener2, hanna wójciak1 1 department of botany and mycology, institute of biology and biochemistry, maria curieskłodowska university, akademicka 19, 20-033 lublin, poland 2 department of geobotany, institute of biology and biochemistry, maria curie-skłodowska university, akademicka 19, 20-033 lublin, poland * corresponding author. email: agata.wolczanska@poczta.umcs.lublin.pl abstract the paper presents new records of three rare species of smut fungi in poland. anthracoidea buxbaumii was collected in new localities, a. caricis collected on carex montana is a new fungus/host combination in poland, and urocystis ranunculiauricomi was found in the country after almost 50 years. keywords urocystales; ustilaginales; anthracoidea; urocystis; distribution in poland introduction smut fungi (ustilaginomycotina) are multicellular organisms characteristic by their dark, thick-walled, and dust-like teliospores. smuts parasitize flowering plants, including many economically important hosts like maize, barley, wheat, oats, and forage grasses. detailed data about the distribution of smut fungi in poland are summarized in the monograph by kochman and majewski [1] and in a preliminary checklist of micromycetes in poland [2]. articles presenting data about the occurrence of this group in the country were published by chlebicki [3], piątek and mułenko [4], ruszkiewicz et al. [5], and lutz and piątek [6]. anthracoidea species are widely distributed in the temperate and subarctic regions of the northern hemisphere and the highland regions of the southern hemisphere. currently, this genus is represented by approximately 112 species and this number is supposed to increase [7–10]. the representatives of urocystis are cosmopolitan and occur worldwide. vanky [11] has recorded as many as 162 species in this genus. the aim of this paper is to present new records of the distribution of some rare smut fungal species from poland, i.e., anthracoidea buxbaumii kukkonen, a. caricis (pers.) bref., and urocystis ranunculi-auricomi (liro) zundel. material and methods infected host plants were collected in the southeastern part of poland: polesie region, lublin upland, środkowomazowiecka lowland, and jasło–krosno basin. air-dried specimens were examined under a standard light microscope olympus ch30 and a scanning electron microscope (sem) vega3 tescan. the identification of the fungi and their nomenclature follow that provided by kochman and majewski [1] and vanky [11]. the names of vascular plants are unified according to mirek et al. [12] and the regions of poland according to kondracki [13]; additionally, data on the atpol squares have been added to each locality. the analyzed specimens are deposited in the herbarium of the department of botany and mycology, maria curie-skłodowska university in lublin (lbl). doi: 10.5586/am.1072 publication history received: 2016-04-12 accepted: 2016-06-15 published: 2016-06-24 handling editor maria rudawska, institute of dendrology, polish academy of sciences, poland authors’ contributions all authors collected the material and contributed to manuscript preparation; aw identified all specimens funding research supported by the polish ministry of science and higher education as part of the statutory activities of the department of botany and mycology and the department of geobotany, maria curieskłodowska university in lublin. competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation wołczańska a, cwener a, wójciak h. addendum to the mycobiota of smut fungi in poland. acta mycol. 2016;51(1):1072. http://dx.doi. org/10.5586/am.1072 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:agata.wolczanska%40poczta.umcs.lublin.pl?subject=addendum%20to%20the%20mycobiota%20of%20smut%20fungi%20in%20poland http://dx.doi.org/10.5586/am.1072 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1072 http://dx.doi.org/10.5586/am.1072 2 of 5© the author(s) 2016 published by polish botanical society acta mycol 51(1):1072 wołczańska et al. / addendum to the mycobiota of smut fungi in poland list of species anthracoidea buxbaumii kukkonen sori globose in the ovaries, usually scattered in the inflorescence, rarely: almost all ovaries infected (fig. 1d). spores dark reddish brown, flattened, in plane view broadly elliptical, ovate or irregular, 14–22(–24) × 20–26(–28) µm; wall 1–3.5 µm thick, with 1–3 indistinct internal swellings, without protuberances and light-refractive areas, minutely verruculose. specimens examined. on carex buxbaumii wahlenb.: poland, volhynian polesie, dubienka depression: ca. 4 km ne of the bagno serebryskie reserve, and 2 km n of the brzeźno reserve (ge 25), 6 june 2015, leg. a. cwener (lbl 23587); western polesie, łęczna–włodawa plain: krowie bagno reserve (ge 02), 3 july 2015, leg. h. wójciak (lbl 23588). notes. in all the localities mentioned above, the host plants were heavily infected. probably in the polish part of the polesie region, a. buxbaumii can be considered a relatively frequent species. two localities in poland situated in this area have been published so far: anthracoidea buxbaumii was found on c. buxbaumii in bagno fig. 1 infected host plants and spores of the presented species. a,b anthracoidea caricis on carex montana. c,e,f urocystis ranunculi-auricomi on ranunculus auricomus (the white circle indicates an infected leaf ). d anthracoidea buxbaumii on carex buxbaumii. 3 of 5© the author(s) 2016 published by polish botanical society acta mycol 51(1):1072 wołczańska et al. / addendum to the mycobiota of smut fungi in poland serebryskie reserve near chełm town (ge 34) and on c. hartmanii cajander in the bagno bubnów swamp (ge 12) in the poleski national park [4,14] (fig. 2). the parasite forms black spore clusters on female inflorescences of c. buxbaumii and c. hartmanii sedges. these plants represent rare taxa in poland and occur in scattered localities over the country. previously, they were regarded as an aggregate unit [15]. currently, accurate data on their occurrence as separate species have been presented by sotek [16,17] and gierczyk and soboń [18]. both sedge species occur in peat bogs, i.e., habitats that are decreasing due to the reduced water level. carex buxbaumii is included in the list of endangered taxa in poland [19], which has been proposed to include c. hartmanii as well [17]. in the regional list concerning the polesie and lublin regions, c. buxbaumii agg. is specified as a vulnerable species [20,21]. detailed data on the distribution of a. buxbaumii worldwide are presented in the paper by piątek and mułenko [4]. anthracoidea caricis (pers.) bref. sori black, globose in ovaries. spores middle to dark reddish brown, flattened, in plane view subcircular, angular, or irregular, 16–19(–20) × 18–24 µm; wall 1.5–3 µm thick, the thickest at the angles, with 1–3 indistinct internal swellings, minutely verruculose; besides warts (0.2–0.3 µm high), minute papillae are visible in sem (fig. 1a,b). specimens examined. on carex montana l. poland, lublin upland, zamość depression: surroundings of niedzieliska village near zamość town – edge of bodaczowski forest (ge 91), 19 may 2015, leg. a. cwener (lbl 23586). on c. pilulifera l. poland, środkowomazowiecka lowland, central vistula valley: surroundings of dęblin town (fd 92), july 2015, leg. h. wójciak (lbl 23585). notes. anthracoidea caricis is one the most common species of anthracoidea in poland. until now, it has been collected only on c. pilulifera – data on c. montana [1,2] should be referred to ukraine (pokucie carpathians). this is first collection of the species on c. montana in poland. the general distribution of this species ranges across the northern hemisphere [11]. urocystis ranunculi-auricomi (liro) zundel sori on leaves and stems create up to 2-cm long swellings. initially, they are covered by epidermis; later, after rupturing, black powdery mass is visible (fig. 1e,f ). spore balls are composed of brown central spores (2–7 in one ball) and yellowish sterile cells (3–7). balls: 22–32 × 26–42 µm, central spores: 10–16 × 14–18 µm, sterile cells: 4–6 × 8–10 µm (fig. 1c). these data differ only slightly from the species description given by vanky [11]. specimens examined. on ranunculus auricomus l. poland, central beskidian piedmont, jasło–krosno basin: rymanów town (fg 13), wet meadow, 26 april 2011, 23 april 2014, leg. a. wołczańska (lbl 23583, 23584). a b c d e f g a b c d e f g g f e d c b a g f e d c b a a1 a2 b1 b 2 fig. 2 distribution of the analyzed species in poland: anthracoidea buxbaumii: a1 – new localities, a2 – known localities; urocystis ranunculi-auricomi: b1 – new locality, b2 – known locality. 4 of 5© the author(s) 2016 published by polish botanical society acta mycol 51(1):1072 wołczańska et al. / addendum to the mycobiota of smut fungi in poland notes. currently, 32 species of the urocystis genus occur in poland. three species, i.e., u. ficariae (liro) moesz, u. ranunculi (lib.) moesz, and u. ranunculi-auricomi [2], have been found on representatives of the ranunculus genus. in turn, nine species have been reported on this host worldwide [11]. urocystis ranunculi is the most common cosmopolitan species, whereas the other species are only distributed across the northern hemisphere. urocystis ranunculi-auricomi is quite a rare species known in europe, asia, and north america. it was collected most frequently on ranunculus auricomus and rarely on r. aconitifolius l., r. affinis r. br., r. cassubicus l., r. escholtzii schltdl., r. fallax (wimmer & grab.) kerner, r. monophyllus ovcz, and r. sibiricus glehn [11]. in poland, this rare species has been reported from one locality so far, i.e., from stanisławice village in niepołomicka primeval forest (ef 63). the specimen was collected by j. kućmierz on a wet meadow in may 1964 [1,22] (fig. 2). data from ostrowiec published by kawecka-starmachowa [23] refer to an area that currently belongs to ukraine (leg. t. wilczyński in 1913). although the hosts of u. ranunculiauricomi are common in poland, the second locality of this smut fungus was found after nearly 50 years. the r. auricomus specimens found in rymanów were infected by two fungal species. in addition to u. ranunculi-auricomi, aecidium ranunculacearum dc aecia were found on the plant leaves. references 1. kochman j, majewski t. podstawczaki (basidiomycetes), głowniowe (ustilaginales). warszawa: państwowe wydawnictwo naukowe; 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(biodiversity of poland; vol 1). 13. kondracki j. geografia regionalna polski. warszawa: wydawnictwo naukowe pwn; 2009. http://dx.doi.org/10.5586/am.2010.023 http://dx.doi.org/10.5586/am.2010.023 http://dx.doi.org/10.5586/am.2012.026 http://dx.doi.org/10.1007/s10658-016-0874-1 http://dx.doi.org/10.3852/12-074 http://dx.doi.org/10.12664/mycobiota.02.01 http://dx.doi.org/10.12664/mycobiota.02.01 http://dx.doi.org/10.1007/s11557-015-1140-1 http://dx.doi.org/10.11646/phytotaxa.244.1.5 5 of 5© the author(s) 2016 published by polish botanical society acta mycol 51(1):1072 wołczańska et al. / addendum to the mycobiota of smut fungi in poland 14. piątek m, ruszkiewicz-michalska m, mułenko w. catalogue of polish smut fungi, with notes on four species of anthracoidea. pol bot j. 2005;50(1):19–37. 15. zając a, zając m. atlas rozmieszczenia roślin naczyniowych w polsce. kraków: pracownia chorologii komputerowej instytutu botaniki uniwersytetu jagiellońskiego; 2001. 16. sotek z. the distribution of carex buxbaumii wahlenb. in poland. acta soc bot pol. 2006;75(4):293–296. http://dx.doi.org/10.5586/asbp.2006.035 17. sotek z. the distribution of carex hartmanii cajander in poland. acta soc bot pol. 2008;77(4):323–326. http://dx.doi.org/10.5586/asbp.2008.042 18. gierczyk b, soboń j. nowe stanowiska chronionych, zagrożonych i rzadko spotykanych gatunków roślin naczyniowych w polsce. przegląd przyrodniczy. 2008;19(3–4):19–31. 19. zarzycki k, szeląg z. red list of the vascular plants in poland. in: mirek z, zarzycki k, wojewoda w, szeląg z, editors. red list of plant and fungi in poland. cracow: w. szafer institute of botany, polish academy of sciences; 2006. 20. kucharczyk m, wójciak j. ginące i zagrożone gatunki roślin naczyniowych wyżyny lubelskiej, roztocza, wołynia zachodniego i polesia lubelskiego. ochrona przyrody. 1995;52:33–46. 21. kucharczyk m, szukałowicz i. rzadkie i zagrożone gatunki roślin polesia zachodniego. kosmos. 2003;52(2–3):321–330. 22. kućmierz j. o kilku głowniach (ustilaginales) zebranych w południowej polsce. fragmenta floristica et geobotanica. 1966;12(1):115–118. 23. kawecka-starmachowa b. głownie i śniecie polski (materiały do monografii). część 2. śniecie. sprawozdanie komisji fizjograficznej. 1939;73:147–224. http://dx.doi.org/10.5586/asbp.2006.035 http://dx.doi.org/10.5586/asbp.2008.042 abstract introduction material and methods list of species anthracoidea buxbaumii kukkonen anthracoidea caricis (pers.) bref. urocystis ranunculi-auricomi (liro) zundel references 2016-06-24t15:05:49+0100 piotr otręba 2014-11-20t23:22:17+0000 polish botanical society micromycetes on ericaceous plant leaves 1 of 9published by polish botanical society acta mycologica original research paper micromycetes on ericaceous plant leaves maria kowalik*, joanna bonio, klaudia duda-franiak department of plant protection, faculty of biotechnology and horticulture, university of agriculture in krakow, al. 29 listopada 54, 31-425 cracow, poland * corresponding author. email: m.kowalik@ogr.ur.krakow.pl abstract a two-year study was carried out on the ericaceous plant collection of the botanic garden of the jagiellonian university in cracow and the rogów arboretum of the warsaw university of life sciences on the following plants: wild rosemary ledum palustre l., leatherleaf chamaedaphne calyculata (l.) moench and american cranberry oxycoccus macrocarpus (ait.) pursh. diverse micromycetes species composition was specified on the leaves of tested ericaceous plants. the perpetrators of dying leaves were recognized, among which the dominant role was played by the necrotroph pestalotiopsis sydowiana. quantitative and qualitative comparative analyses of micromycetes in both locations were carried out, showing a comparable degree of plant colonization by these fungi in the botanic garden and arboretum. the study may be helpful in explaining the causes of dieback of protected and endangered plants in polish flora. keywords ericaceae; leaves; fungi; botanic garden; arboretum this issue of acta mycologica is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. introduction the role of botanic gardens is to protect rare and endangered plant species, so one of the duties is the storage and reproduction of these species. the following plants are threatened by extinction: wild rosemary ledum palustre l. (syn. rhododendron tomentosum harmaja), leatherleaf chamaedaphne calyculata (l.) moench and american cranberry oxycoccus macrocarpus (ait.) pursh. (syn. vaccinium macrocarpon ait.). wild rosemary grows on peat bogs and swamp forests. this is a medicinal plant, often obtained from natural habitats, although in poland is under partial protection. leatherleaf is the only species in the genus chamaedaphne. in poland its natural habitat is the kampinos national park and drawa national park, where this species is under careful protection as a relic of glaciation. in the current polish red book of plants, ch. calyculata is classified in the group en – endangered. the american cranberry grows in natural habitats and wetlands in the eastern part of north america but in poland is recognized as domesticated plant. these three plant species transferred to the collection of the botanic garden of the jagiellonian university in cracow and the rogów arboretum of the warsaw university of life sciences, showed symptoms of poor health and revealed extensive necrosis on leaves. it resulted in leaf dieback and defoliation, and further dieback of sprouts and shrubs. leatherleaf and wild rosemary were especially affected by a disease. finally wild rosemary plants died in the botanic garden in 2013. doi: 10.5586/am.1055 publication history received: 2015-02-11 accepted: 2015-07-02 published: 2015-08-05 handling editor tomasz leski, institute of dendrology of the polish academy of sciences, poland authors’ contributions mk, jb: concept of the study; kdf, mk: determination of the specimens; mk, kdf: writing the manuscript funding research supported by the polish ministry of science and higher education as part of the statutory activities of the department of plant protection, university of agriculture in krakow. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation kowalik m, bonio j, dudafraniak k. micromycetes on ericaceous plant leaves. acta mycol. 2015;50(1):1055. http:// dx.doi.org/10.5586/am.1055 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:m.kowalik%40ogr.ur.krakow.pl?subject=micromycetes%20on%20ericaceous%20plant%20leaves http://dx.doi.org/10.5586/am.1055 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1055 http://dx.doi.org/10.5586/am.1055 2 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1055 kowalik et al. / micromycetes on ericaceous plants this poor condition of plants was the inspiration for starting mycological studies aiming to determine the species composition of micromycetes inhabiting the leaves of l. palustre, ch. calyculata and o. macrocarpus, identification the perpetrators of leaf dieback and finally to conduct a comparative analysis of the mycobiota at these two locations. material and methods the study was carried out in the collection of ericaceous plants of the botanic garden of the jagiellonian university in cracow (cracow botanic garden) and the rogów arboretum of the warsaw university of life sciences (rogów arboretum). mycological analysis included the following plants: wild rosemary l. palustre, leatherleaf ch. calyculata and american cranberry o. macrocarpus. the sources of the research material were leaves of leatherleaf and american cranberry with necrosis, collected in may, july and september 2012 and 2013 and wild rosemary in the same months in 2012. micromycetes were isolated from a total of 120 wild rosemary leaf fragments and 240 leaf fragments of leatherleaf and american cranberry. on both tested sites twenty leaves from each plant species were collected at every time point. leaf fragments were cut from the border of healthy and necrotic tissues from single spots and surface decontaminated in 70% ethanol for minute, then thoroughly rinsed three times for one minute in sterile water before being placed on a petri dish with a 2% pda medium. the petri dishes were incubated for 7 days at 21–22°c. the microscope used for observation was a delta optical microscope, model evolution 300. cultures of representative species of fungi are held in the department of plant protection of the university of agriculture in krakow. the taxonomic identification of micromycetes was conducted after: domsch et al. [1], ellis and ellis [2], guba [3], and sutton [4]. the basis for the classification was the kirk et al. [5] system, and author citations for each taxon are abbreviated according to the index fungorum database accessed november 2014 [6]. the similarity coefficient was calculated for the micromycetes communities isolated from the leaves of the tested plants, including habitats and years of research according to the formula given by błaszkowski et al. [7]: s = w / a + b − w, where: s – similarity of compared communities (range of coefficient variation 0–1), a, b – number of species in communities a, b, w – number of shared species in both communities. results micromycetes communities inhabiting the leaves of wild rosemary, leatherleaf and american cranberry growing in the cracow botanic garden and the rogów arboretum differed in species composition and number of isolated colonies. in total 583 micromycetes colonies, comprising 31 species, were isolated from the leaves of ericaceous plants. one hundred and sixty micromycetes colonies isolated from 120 infected wild rosemary leaves fragments resulted in identification of 15 species (tab. 1). the dominant fungi were: pestalotiopsis sydowiana (35% of all of the fungi communities), alternaria alternata and epicoccum nigrum. the leaves of wild rosemary from the botanic garden and arboretum were inhabited by fungi in a comparable rates. seventy-eight isolates belonging to 9 species and 82 isolates representing 11 species were isolated from the botanic garden and arboretum, respectively. five species – a. alternata, e. nigrum, penicillium expansum, p. sydowiana and phialophora cyclaminis – were common for both tested sites. from 187 colonies of micromycetes, isolated from the dying leaves of leatherleaf, 18 species has been determined (tab. 2). pestalotiopsis sydowiana played a major role (as well as on wild rosemary) and constituted almost 30% of the entire fungi community. pestalotia rhododendri, a. alternata, e. nigrum, umbelopsis isabellina, sordaria fimicola, ph. cyclaminis and phoma medicaginis were also numerous. their participation in the community of identified fungi was above 57%. the similar number of 3 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1055 kowalik et al. / micromycetes on ericaceous plants ta b. 1 m ic ro m yc et es o n le av es o f w ild ro se m ar y le du m p al us tr e l. fu ng us fu ng i f re qu en cy o n le av es in : to ta l pe rc en ta ge (% ) c ra co w b ot an ic g ar de n r og ów a rb or et um 20 12 20 12 m ay ju ly se pt em be r m ay ju ly se pt em be r pe st al ot io ps is sy do w ia na (b re s. ) b . s ut to n 6 15 13 10 2 10 56 35 .0 0 a lte rn ar ia a lte rn at a (f r.) k ei ss l. 4 1 5 6 16 10 .0 0 ep ic oc cu m n ig ru m l in k 2 4 2 4 2 2 16 10 .0 0 pe ni ci lli um e xp an su m l in k 2 9 11 6. 88 ph ia lo ph or a cy cl am in is j. f. h . b ey m a 6 4 10 6. 25 ph om a m ed ic ag in is m al br . & r ou m . 6 3 9 5. 63 le pt os ph ae ri a m ac ul an s ( tu l.) c es . & d e n ot . 8 8 5. 00 so rd ar ia fi m ic ol a (r ob er ge e x d es m .) c es . & d e n ot . 4 2 6 3. 75 fu sa ri um fl oc ci fe ru m c or da 6 6 3. 75 fu sa ri um o xy sp or um s ch ltd l. 6 6 3. 75 m or tie re lla a lp in a pe yr on el 5 1 6 3. 75 a cr em on ie lla a tr a (c or da ) s ac c. 4 4 2. 50 fu sa ri um c ul m or um (w m .g . s m .) sa cc . 2 2 1. 25 u m be lo ps is is ab el lin a (o ud em .) w . g am s 2 2 1. 25 m or tie re lla p ar vi sp or a li nn em . 2 2 1. 25 to ta l i n m on th 12 20 46 32 16 34 to ta l 78 82 16 0 10 0. 00 4 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1055 kowalik et al. / micromycetes on ericaceous plants ta b. 2 m ic ro m yc et es o n le av es o f l ea th er le af c ha m ae da ph ne c al yc ul at a (l .) m oe nc h fu ng us fu ng i f re qu en cy o n le av es in : to ta l pe rce nt ag e (% ) c ra co w b ot an ic g ar de n r og ów a rb or et um 20 12 20 13 20 12 20 13 m ay ju l se p m ay ju l se p m ay ju l se p m ay ju l se p pe st al ot io ps is sy do w ia na (b re s. ) b . s ut to n 2 3 7 7 5 10 14 5 3 56 29 .9 5 pe st al ot ia r ho do de nd ri (d . s ac c. ) g ub a 5 7 4 5 21 11 .2 3 a lte rn ar ia a lte rn at a (f r.) k ei ss l. 4 5 1 4 6 20 10 .7 0 ep ic oc cu m n ig ru m l in k 6 1 1 2 2 2 1 2 17 9. 09 u m be lo ps is is ab el lin a (o ud em .) w . g am s 2 5 10 17 9. 09 so rd ar ia fi m ic ol a (r ob er ge e x  d es m .) c es . &  d e n ot . 5 2 3 1 2 13 6. 95 ph ia lo ph or a cy cl am in is j. f. h . b ey m a 6 3 1 10 5. 35 ph om a m ed ic ag in is m al br . &  r ou m . 4 1 4 1 10 5. 35 pa ra ph om a ch ry sa nt he m ic ol a (h ol ló s) g ru yt er , a ve ska m p &  v er kl ey 4 1 5 2. 67 tr ic ho de rm a ps eu do ko ni ng ii r ifa i 4 4 2. 14 c ha et om iu m g lo bo su m k un ze 3 3 1. 60 pe ni ci lli um e xp an su m l in k 1 2 3 1. 60 a sp er gi llu s v er si co lo r (v ui ll. ) t ir ab . 1 1 2 1. 07 h um ic ol a fu sc oa tr a tr aa en 2 2 1. 07 bo er em ia e xi gu a (d es m .) a ve sk am p, g ru yt er & v er kl ey 1 1 0. 53 bo tr yt is c in er ea p er s. 1 1 0. 53 sc le ro tin ia sc le ro tio ru m (l ib .) de b ar y 1 1 0. 53 tr ic ho de rm a ha rz ia nu m r ifa i 1 1 0. 53 to ta l i n m on th 22 17 10 15 12 17 21 10 19 9 15 20 to ta l 49 44 50 44 18 7 10 0. 00 5 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1055 kowalik et al. / micromycetes on ericaceous plants colonies was isolated from leaves of leatherleaf and 17 and 10 species were identified from the botanic garden and from the arboretum respectively. nine fungi species were common for leatherleaf leaves in both sites. two hundred thirty six micromycetes colonies were isolated from infected and brown american cranberry leaf tissues, including 16 species (tab. 3). the dominant fungus was p. sydowiana, which accounted for more than 63% of all of the colonies. other species found in large number of colonies were p. rhododendri and a. alternata, while fungi from giberella, epicoccum, phialophora and sordaria types appeared in the lower abundance. pestalotiopsis sydowiana, a. alternata, p. rhododendri, giberella stilboides and e. nigrum were common species for both locations. a greater diversity of fungi species was found in the phyllosphere of american cranberry in the cracow botanic garden than in the rogów arboretum. only four species – a. alternata, e. nigrum, p. sydowiana and p. rhododendri – inhabited the leaves of the ericaceous plants during both years of the study in the botanic garden and the arboretum. aspergillus versicolor, boeremia exigua, botrytis cinerea, davidiella macrocarpa, humicola fuscoatra, sclerotinia sclerotiorum and trichoderma harzianum were occasionally found on the ericaceous plants. the similarity between the studied micromycetes communities (within locations and year of study) ranged from 0.1 to 0.88 (tab. 4). the lowest rate of similarity was calculated for the micromycetes communities inhabiting the leaves of leatherleaf in the botanic garden and american cranberry in the arboretum in 2012. micromycetes inhabiting leatherleaf and american cranberry in both locations and years as well as wild rosemary and leatherleaf in the botanic garden were also characterized by low similarity coefficients. the highest similarity coefficient (0.88) was calculated for the micromycetes communities on leatherleaf leaves from the botanic garden and arboretum in 2012. a slightly lower similarity coefficient (0.55) was found for american cranberry in both locations in 2013. discussion there is little information in the literature about the diseases, their potential causes and the fungi inhabiting the leaves of l. palustre, ch. calyculata and o. macrocarpus [8–11]. the phyllosphere of ericaceous plants in the cracow botanic garden and the rogów arboretum was dominated by p. sydowiana (syn. pestalotia sydowiana, pestalotia guepini var. rhododendri). this fungus is also mentioned in other papers as common on the leaves of rhododendron, gaultheria, epigaea, oxycoccus, kalmia, calluna, and erica [2–4,12–19]. the previous research on the causes of leatherleaf and wild rosemary leaf death indicates on rust fungi: chrysomyxa ledi, ch. cassandrae, ch. vaccinii and ch. reticulata [10]. however in our studies we did not isolated these fungi, and symptoms of rust on plants grown in the botanic garden and arboretum were not observed. bristow and windom [8], caruso and ramsdell [9] and polashock [20] write about cranberry as susceptible to pathogen infections of leaves, sprouts, fruits and roots. american cranberry in its natural habitats is infected by monilinia oxycocci, lophodermium oxycocci, exobasidium oxycocci, phomopsis vaccinii, fusicoccum putrfaciens, glomerella cingulata, coleophoma empetri, botryosphaeria vaccinii and phyllosticta elongata [11,21,22]. caruso et al. [23] ascribe a special role to the genera phomopsis, fusicoccum, colletotrichum, gloeosporium, aureobasidium and pestalotia in the yellowing and falling leaves, and caruso and ramsdell [9] provide information about the fungi protoventuria myrtilli, pyrenobotrys compacta, exobasidium rostrupii and e. perenne on low-lying leaves of cranberries. in the present study, there was no incidence of these pathogens on o. macrocarpus in the botanic garden and arboretum. a comparative analysis of fungal communities indicate that the species a. alternata, b. cinerea, e. nigrum, p. sydowiana, p. rhododendri, ph. cyclaminis, ph. medicaginis and s. fimicola (with a high frequency) and numerous species belonging to 6 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1055 kowalik et al. / micromycetes on ericaceous plants ta b. 3 m ic ro m yc et es o n le av es o f a m er ic an c ra nb er ry o xy co cc us m ac ro ca rp us (a it. ) p ur sh . fu ng us fu ng i f re qu en cy o n le av es in : to ta l pe rce nt ag e (% ) c ra co w b ot an ic g ar de n r og ów a rb or et um 20 12 20 13 20 12 20 13 m ay ju l se p m ay ju l se p m ay ju l se p m ay ju l se p pe st al ot io ps is sy do w ia na (b re s. ) b . s ut to n 13 5 15 25 18 14 16 16 13 15 15 0 63 .5 6 a lte rn ar ia a lte rn at a (f r.) k ei ss l. 3 2 1 1 3 1 2 1 1 2 17 7. 20 pe st al ot ia r ho do de nd ri (d . s ac c. ) g ub a 4 2 5 3 14 5. 93 ep ic oc cu m n ig ru m l in k 1 2 3 1 2 9 3. 81 g ib be re lla st ilb oi de s w .l . g or do n ex c . b oo th 1 3 3 7 2. 97 ph ia lo ph or a cy cl am in is j. f. h . b ey m a 1 3 3 7 2. 97 so rd ar ia fi m ic ol a (r ob er ge e x  d es m .) c es . &  d e n ot . 5 5 2. 12 u m be lo ps is is ab el lin a (o ud em .) w . g am s 2 3 5 2. 12 bo er em ia e xi gu a (d es m .) a ve sk am p, g ru yt er &  v er kl ey 1 3 4 1. 69 ph om a le ve ill ei b oe re m a &  g .j. b ol le n 4 4 1. 69 c la do sp or iu m c la do sp or io id es (f re se n. ) g .a . d e  v ri es 1 2 3 1. 27 ph om a m ed ic ag in is m al br . &  r ou m . 3 3 1. 27 tr ic ho de rm a ha m at um (b on or d. ) b ai ni er 3 3 1. 27 pa ra ph om a ch ry sa nt he m ic ol a (h ol ló s) g ru yt er , a ve ska m p &  v er kl ey 2 2 0. 85 ph ia lo ph or a ci ne re sc en s ( w ol le nw .) j.f .h . b ey m a 1 1 2 0. 85 d av id ie lla m ac ro ca rp a c ro us , k . s ch ub . & u . b ra un 1 1 0. 42 to ta l i n m on th 22 16 21 6 33 37 15 18 16 12 21 19 to ta l 59 76 49 52 23 6 10 0. 00 7 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1055 kowalik et al. / micromycetes on ericaceous plants the genera chaetomium, cladosporium, davidiella, fusarium, mortierella, umbelopsis and trichoderma were common fungi inhabiting the leaves of azalea, evergreen rhododendron, wild rosemary, leatherleaf and american cranberry in previous vegetation periods in the cracow botanic garden [12,13,15,16]. the research carried out in the rogów arboretum 2011 [14] shows the similar results. the previous results showed that a dozen micromycetes species were common for the leaves and flowers of saucer magnolia in the botanic garden in 2013 [24] and for the leaves of ericaceous plants. alternaria alternata, e. nigrum, mortierella parvispora, p. expansum, ph. medicaginis inhabited the infected organs with a high frequency. comparison of mycological analysis of air carried out in the botanic garden and arboretum in 2012 [25] with the fungal communities inhabiting the ericaceous plants revealed very low similarity in terms of colony-forming units for the air and tested plants. the air of the botanic garden and the leaves of wild rosemary, leatherleaf and american cranberry contained a. alternata, e. nigrum, p. expansum and c. cladosporioides, while the air of the arboretum and the ericaceous plants contained three species: a. alternata, e. nigrum and p. expansum. additionally f. oxysporum was found on the leaves of wild rosemary. the low and very low similarity coefficients calculated for the micromycetes communities inhabiting wild rosemary, leatherleaf and american cranberry in the two years of research and locations is an evidence of the diverse species composition in the phyllosphere of ericaceous plants. conclusions (i) micromycetes communities inhabiting the leaves of wild rosemary, leatherleaf and american cranberry , growing in the cracow botanic garden and the rogów arboretum, differed in their species composition and the number of colonies. (ii) the dieback of wild rosemary, leatherleaf and american cranberry was caused by a variety of micromycetes, among which necrotroph p. sydowiana played a dominant role. tab. 4 similarity coefficient of the micromycetes communities isolated from the leaves of ericaceous plants (bg – cracow botanic garden; a – rogów arboretum). species oxycoccus macrocarpus chamaedaphne calyculata ledum palustre year 2012 2013 2012 2013 2012 stand bg a bg a bg a bg a bg a o xy co cc us m ac ro ca rp us 2 01 2 bg 0.22 0.31 0.20 0.39 0.36 0.18 0.25 0.39 0.33 a 0.22 0.25 0.22 0.10 0.14 0.18 0.33 0.22 0.18 20 13 bg 0.31 0.25 0.55 0.21 0.27 0.27 0.40 0.21 0.27 a 0.20 0.22 0.55 0.20 0.15 0.43 0.50 0.29 0.25 c ha m ae da ph ne ca ly cu la ta 20 12 bg 0.39 0.10 0.21 0.20 0.88 0.18 0.25 0.20 0.43 a 0.36 0.14 0.27 0.15 0.88 0.13 0.20 0.25 0.42 20 13 bg 0.18 0.18 0.27 0.43 0.18 0.13 0.42 0.18 0.22 a 0.25 0.33 0.40 0.50 0.25 0.20 0.42 0.36 0.42 le du m pa lu st re 20 12 bg 0.39 0.22 0.21 0.29 0.20 0.25 0.18 0.36 0.33 a 0.33 0.18 0.27 0.25 0.43 0.42 0.22 0.42 0.33 8 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1055 kowalik et al. / micromycetes on ericaceous plants references 1. domsch kh, gams w, anderson th. compendium of soil fungi. london: academic press; 1980. 2. ellis mb, ellis jp. microfungi on land plants. an identification handbook. london: croom helm; 1987. 3. guba ef. monograph of monochaetia and pestalotia. cambridge, ma: harvard university press; 1961. 4. sutton bc. the coelomycetes. fungi imperfecti with pycnidia, acervuli and stromata. kew: commonwealth mycological institute; 1980. 5. kirk pm, cannon pf, minter dw, stalpers ja. ainsworth & bisby’s dictionary of the fungi. 10th ed. wallingford: cabi; 2008. 6. index fungorum [internet]. 2014 [cited 2014 nov 14]; available from: http://www.indexfungorum.org 7. błaszkowski j, tadych m, madej t. przewodnik do zajęć z fitopatologii. szczecin: wydawnictwo akademii rolniczej w szczecinie; 1999. 8. bristow pr, windom ge. controlling twig blight and fruit rots of cranberry with fungicides. fungicide and nematicide tests. 1989;44:81. 9. caruso fl, ramsdell dc. compendium of blueberry and cranberry diseases. st. paul, mn: aps press; 1995. 10. crane pe. morphology, taxonomy, and nomenclature of the chrysomyxa ledi complex and related rust fungi on spruce and ericaceae in north america and europe. can j bot. 2001;79:957–982. http://dx.doi.org/10.1139/cjb-79-8-957 11. oudemans pv, caruso fl, stretch aw. cranberry fruit rot in the northeast: a complex disease. plant dis. 1998;82:1176–1184. http://dx.doi.org/10.1094/pdis.1998.82.11.1176 12. kowalik m. fungi and fungi-like oomycetes isolated from affected leaves of rhododendron. acta mycol. 2008;43(1):21–27. http://dx.doi.org/10.5586/am.2008.003 13. kowalik m. bioróżnorodność grzybów występujących w fyllosferze różanecznika zimozielonego rhododendron l. zeszyty problemowe postępów nauk rolniczych. 2009;539:341–348. 14. kowalik m. diversity of fungi colonizing and damaging leaves of pontic azalea azalea pontica l. acta mycol. 2013;48(2):227–236. http:/dx.doi.org/10.5586/am.2013.024 15. kowalik m, kierpiec b, bonio j, żołna m. fungi inhabiting spots and necroses on the leaves of azaleas (rhododendron) in the botanical garden of the jagiellonian university. phytopathologia. 2011;62:41–48. 16. kowalik m, kierpiec b, żołna m. fungi living at the fallen leaves of rhododendron and azalea (rhododendron l.) acta sci pol hortorum cultus. 2012;11(2):161–166. 17. kowalik m, wandzel a. grzyby powodujące zamieranie sadzonek wrzosu. acta agrobot. 2005;58(2):237–242. http://dx.doi.org/10.5586/aa.2005.050 18. kowalik m, wilczek e. grzyby powodujące zamieranie sadzonek wrzośca erica l. prog plant prot post ochr roślin. 2008;48(1):199–203. 19. kowalik m, wójcik j. grzyby powodujące zamieranie pędów wrzosu na terenie ogrodu botanicznego uniwersytetu jagiellońskiego. prog plant prot post ochr roślin. 2009;49(1):214–218. 20. polashock jj, caruso fl, oudemans pv, mcmanus ps, crouch ja. the north american cranberry fruit rot fungal community: a systematic overview using morphological and phylogenetic affinities. plant pathol. 2009;58(6):1116–1127. http://dx.doi. org/10.1111/j.1365-3059.2009.02120.x 21. jeffers s. seasonal incidence of fungi in symptoms less cranberry leaves and fruit (iii) most of the micromycetes communities inhabiting the leaves of ericaceous plants in the cracow botanic garden and the rogów arboretum were characterized by a low and very low coefficient of similarity. http://www.indexfungorum.org http://www.indexfungorum.org http://dx.doi.org/10.1139/cjb-79-8-957 http://dx.doi.org/10.1094/pdis.1998.82.11.1176 http://dx.doi.org/10.5586/am.2008.003 http:/dx.doi.org/10.5586/am.2013.024 http://dx.doi.org/10.5586/aa.2005.050 http://dx.doi.org/10.1111/j.1365-3059.2009.02120.x http://dx.doi.org/10.1111/j.1365-3059.2009.02120.x 9 of 9© the author(s) 2015 published by polish botanical society acta mycol 50(1):1055 kowalik et al. / micromycetes on ericaceous plants treated with fungicides during bloom. phytopathology. 1991;81:636–644. http://dx.doi. org/10.1094/phyto-81-636 22. mcmanus ps, best vm, voland rp, leininger bl. sensitivity of monilinia oxycocci to fenbunconazole and propiconazole in vitro and control of cranberry cottonball in the field. plant dis. 1999;83:445–450. http://dx.doi.org/10.1094/pdis.1999.83.5.445 23. caruso fl, bristow pr, oudemans pv. cranberries. the most intriguing native north american fruit. apsnet features. 2000. http://dx.doi.org/10.1094/apsnetfeature-2000-1100 24. duda k, bonio j. grzyby bytujące na magnolii soulange’a (magnolia ×soulangeana soul.bod.) w ogrodzie botanicznym uniwersytetu jagiellońskiego w krakowie. episteme. 2014;3(22):41–47 25. bonio j, duda k. zanieczyszczenia mykologiczne powietrza w ogrodzie botanicznym uniwersytetu jagiellońskiego w krakowie i arboretum szkoły głównej gospodarstwa wiejskiego w rogowie. episteme. 2014;2(22):17–24. http://dx.doi.org/10.1094/phyto-81-636 http://dx.doi.org/10.1094/phyto-81-636 http://dx.doi.org/10.1094/pdis.1999.83.5.445 http://dx.doi.org/10.1094/apsnetfeature-2000-1100 abstract introduction material and methods results discussion conclusions references 2015-08-02t15:53:39+0100 piotr otręba maria lisiewska and anna bujakiewicz – on the occasion of their 80th and 75th birthdays and the 60th anniversary of creating poznań mycocoenological school 1 of 4published by polish botanical society acta mycologica editorial maria lisiewska and anna bujakiewicz – on the occasion of their 80th and 75th birthdays and the 60th anniversary of creating poznań mycocoenological school maria ławrynowicz* department of algology and mycology, faculty of biology and environmental protection, university of łódź, banacha 12/16, 90-237 łódź, poland * email: miklaw@biol.uni.lodz.pl abstract birthday anniversaries of excellent mycologists professor maria lisiewska (born 2 january 1934) and professor anna bujakiewicz (born 1 february 1940) are an occasion for dedicating them the jubilee issue of acta mycologica. both ladies are known as authors of remarkable papers on mycocoenology, chorology and taxonomy of fungi. specially appreciated is their educational activity in the form of courses and seminars opened for people from the whole country. the entity of their work resulted with the creation of poznań mycocoenological school and was important for integration of polish mycologists, which later contributed to establishing the polish mycological society. keywords jubilee; mycocoenology; scientific curriculum; mycological education this issue of acta mycologica is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. dedicating the exceptional 50-th issue of acta mycologica to professors maria lisiewska and anna bujakiewicz is a sign of homage, admiration and gratitude for their over half of a century work in fields of fungal ecology, chorology and taxonomy. their activity in science and education as well as openness for home and abroad cooperation had and still has a huge impact in growth of mycocoenology, integration and exchange of ideas among mycologists. the entity of their work and the means to realize it had gained the designation poznań mycocoenological school. the jubilee ladies undertook mycological studies in 50-ties and 60-ties of the passed century. at that time, there was a vast need to gather macromycetes materials for taxonomy studies of fungi and enriching the phytosociological studies with the presence of fungi in plant communities. professor andrzej nespiak [1,2] carried out such mycological studies in białowieski national park for the first time in poland. the concept of mycocoenology found favorable conditions in the university of adam mickiewicz in poznań, where there was already a greatly advanced phytosociology developed by such known professors as j. paczoski (1864–1942), t. wojterski (1922– 1991), f. celiński (1924–2001), t. krotoska (1927–2006), h. piotrowska (1925) and others. professor z. czubiński (1912–1967), who was head of department of systematic and plant geography uam encouraged including mushrooms in phytocoenological studies. mycocoenological field studies on permanent plots have a high documentary value, allowing observing of changes in ecosystems during long time periods. the gathered herbarial material is a source of information concerning occurrence and doi: 10.5586/am.1062 publication history received: 2015-06-27 accepted: 2015-07-23 published: 2015-08-05 handling editor maria rudawska, institute of dendrology of the polish academy of sciences, poland funding this work did not involve any funding. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation ławrynowicz m. maria lisiewska and anna bujakiewicz – on the occasion of their 80th and 75th birthdays and the 60th anniversary of creating poznań mycocoenological school. acta mycol. 2015;50(1):1062. http:// dx.doi.org/10.5586/am.1062 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:miklaw%40biol.uni.lodz.pl?subject=maria%20lisiewska%20and%20anna%20bujakiewicz%20%e2%80%93%20on%20the%20occasion%20of%20their%2080th%20and%2075th%20birthdays%20and%20the%2060th%20anniversary%20of%20creating%20pozna%c5%84%20mycocoenological%20school http://dx.doi.org/10.5586/am.1062 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1062 http://dx.doi.org/10.5586/am.1062 2 of 4© the author(s) 2015 published by polish botanical society acta mycol 50(1):1062 ławrynowicz / jubilee of professors maria lisiewska and anna bujakiewicz geographical distribution of macromycetes, and preserves it throughout centuries for further studies. it also is the basis for evaluation of threats and verifying red listed fungi. the accomplishments of mycological research in poznań came to light in the summarization of mycocoenological studies in poland on the fifty years anniversary (1952–2002) [3], in which papers by maria lisiewska and anna bujakiewicz were a major contribution. the anniversary ladies dedicated their professional and personal lives to mycology on local, national and international scale. their study areas were both natural and anthropogenic sites. both ladies devoted most of their scientific work to macromycetes of wielkopolska, but they also worked in other regions of poland, and conducted some of the studies together for example in słowiński national park [4] and dębina nature reserve [5]. the celebrated ladies also participated in important team study projects. one of them was the crypto program, the results of which are contained in a series of seven publications in phytocoenosis (1995–1997). maria lisiewska participated in taxonomic work for preparing the series flora polska – grzyby (mycota) [flora of poland: fungi (mycota)], vol. xvii on genus mycena [6], earlier on taking part in field trips for gathering mycological material. anna bujakiewicz has special achievements in mycocoenological studies in phytocoenosis of riparian forests, alder swamps and mountain plant communities. she spend years studying fungi of mt. babia góra. her postdoctoral dissertation “grzyby babiej góry ii. wartość wskaźnikowa macromycetes w zespołach leśnych [fungi of mt. babia góra ii. the indicating value of macromycetes in forest associations]” includes the most important results of these studies. these results were presented on the vii congress of european mycologists which took place in september 1978 in budapest. in the aftermath of this event, anna bujakiewicz received invitations from mycologists from united states, sweden, norway, finland, the netherlands and many others, for leading field studies, seminars, lectures and joined studies and discussions. in turn mycologist from abroad visited poland and with anna bujakiewicz took field and laboratory studies on mycocoenology. three international field sessions were conducted: danish–polish (1981), dutch–swedish–polish (1986) and norwegian– polish (1991). among her publications a paper concerning alder and alluvial forests in in europe and north america holds a special importance [7]. maria lisiewska took part in polish–czech–italian project “mycological monitoring in european oak forests” (1994–1996) describing changes in the species diversity of macromycetes after 20 years. she also participated in projects in germany, denmark and countries of former yugoslavia, e.g., studies in plitvička jezera national park [8]. she presented her results on international congresses, conferences and symposia. in year 1992 she was invited as visiting professor to university of bologna. her distinguish accomplishment was a depiction of fungi in beech forests in a postdoctoral dissertation “grzyby wyższe lasów bukowych we wschodniej części zasięgu buka w europie [macromycetes of beech forests within the eastern part of the fagus area in europe]”. both ladies were invited by professor wulfard winterhoff to prepare chapters in the handbook fungi in vegetation science [9,10]. the mycological research center in poznań is distinguished by its activity on behalf of social education. since 1957 maria lisiewska has been giving lectures and lead field trips focused on identification of mushrooms approved for trade; as a part of annual courses organized in different parts of poland by the provincial sanitary and epidemiological station (sanepid) in poznań. in cooperation with marian szmid from sanepid in poznań she prepared and published five editions of popular book przewodnik grzyboznawczy [guide to mushrooms]. anna bujakiewicz is chairperson to the mycological section of the poznań unit of polish botanical society (pbs) since its establishment in 1991. within it and for 25 years now, she has been organizing nationwide mycological seminars on reviews, new trends and newest achievements in mycology. these monthly, open meetings have a great importance in integrating professional and amateur mycologists. there are specially valued by young people, inspired to take on scientific research and consulting their efforts. in year 2000 an article was published which included a tabular register of 98 reports presented by speakers from 17 research centers in poland [11]. till now more than 200 reports were presented on these seminars. as a part of activity 3 of 4© the author(s) 2015 published by polish botanical society acta mycol 50(1):1062 ławrynowicz / jubilee of professors maria lisiewska and anna bujakiewicz of mycological section of the poznań unit of pbs, prof. bujakiewicz organized for many years mycological field session, often linked with mushroom exhibition, in the regions of notecka (obrzycko; fig. 1) or zielonka forest. the broad range of topics, participation of amateurs, some of who have the knowledge and scientific achievements and are eager to exchange information have shown the need for a larger platform for communication. the issue was discussed on the plenary meeting of the general mycological section chaired by professor alina skirgiełło, during the 52nd pbs convention in poznań on 25 september 2001. a proposal was brought forth to create a society that would bring together not only mycologists with a botanical background, but also mycologists engaged in fields of medicine, veterinary, construction, commercial use of fungi, toxicology, nutrition etc. the proposal was endorsed by the gathered members. compiling the concept of the new society was entrusted to a group of young people. the preparations took time, but these strengthen the conviction that such a unit, that would embrace all the aspects of mycology is needed. after 8 years of discussion and arrangements, the concept of polish mycological society had emerged on the nationwide mycological symposium on 12 september 2009 in olsztyn and the society itself was created on the nationwide polish mycologists convention on november 5th 2011 in łódź. today, the mycological section of pbs and the polish mycological society are connected by a constructive cooperation, especially that some of the members belong to both organizations. the conference “fungi – key players in ecosystem functions” (23–27 september 2014 łódź–spała, poland), was a clear sign of this cooperation. during the conference a special session on ecology of fungi was dedicated to both anniversary ladies. it finished with an excellent lecture by anna bujakiewicz “fungi and plants – curious relations” [12]. later on, during the evening ceremony the anniversary ladies received many congratulations, thanks and greetings. a special presentation by maria ławrynowicz on the 4th congress of european mycologists in warsaw 1966 was dedicated to both ladies who participated in it and maria lisiewska was among organizers. fig. 1 the second field session of the mycological section of the poznań unit of polish botanical society, held in obrzycko near wronki in 1992. 1 – professor maria lisiewska; 2 – professor anna bujakiewicz. midmost (3) professor alina skirgiełło, founder of acta mycologica and its multiannual editor-in-chief (phot. a. łempicki). 4 of 4© the author(s) 2015 published by polish botanical society acta mycol 50(1):1062 ławrynowicz / jubilee of professors maria lisiewska and anna bujakiewicz acknowledgments special thanks to małgorzata połatyńska for her help in preparing the english version of the text. references 1. nespiak a. grzyby kapeluszowe w zespołach leśnych puszczy białowieskiej (komunikat wstępny). fragm flor geobot. 1956;2(2):134–145. 2. nespiak a. studia nad udziałem grzybów kapeluszowych w zespołach leśnych na terenie białowieskiego parku narodowego. warszawa: pwn; 1959. (monographiae botanicae; vol 8). 3. ławrynowicz m, bujakiewicz a, mułenko w. mycocoenological studies in poland – 1952– 2002. łódź: polish botanical society; 2004. (monographiae botanicae; vol 93). http:// dx.doi.org/10.5586/mb.2004.001 4. bujakiewicz a, lisiewska m. mikoflora zbiorowisk roślinnych słowińskiego parku narodowego. bad fizjogr pol zach ser b. 1983;34:49–77. 5. lisiewska m, bujakiewicz a. grzyby wyższe na tle zespołów leśnych. bad fizjogr pol zach ser b. 1976;29:57–67. 6. lisiewska m. podstawczaki (basidiomycetes), bedłkowe (agaricales), gąskowate i (tricholomataceae), grzybówka (mycena). warszawa: pwn; 1987. (flora polska. rośliny zarodnikowe polski i ziem ościennych; vol 17). 7. bujakiewicz a. macrofungi in the alder and alluvial forests in various parts of europe and north america. opera botanica. 1989;100:29–41. 8. lisiewska m, tortič m. macrofungi of forest associations of the plitvička jezera national park (croatia, yugoslavia). acta bot croat. 1990;49:81–91. 9. bujakiewicz a. macrofungi on soil in deciduous forests. in: winterhoff w, editor. fungi in vegetation science. dordrecht: kluwer academic publishers. 1992. p. 49–78. (handbook of vegetation science; vol 19). http://dx.doi.org/10.1007/978-94-011-2414-0 10. lisiewska m. macrofungi on special substrates. in: winterhoff w, editor. fungi in vegetation science. dordrecht: kluwer academic publishers. 1992. p. 151–182. (handbook of vegetation science; vol 19). http://dx.doi.org/10.1007/978-94-011-2414-0. 11. bujakiewicz a. z życia sekcji mikologicznej przy oddziale poznańskim polskiego towarzystwa botanicznego. problemy społecznego ruchu naukowego. 2000;6(4):245–256. 12. ruszkiewicz-michalska m, szkodzik j, editors. fungi – key players in ecosystem functions. book of abstracts. warszawa: polskie towarzystwo mykologiczne; 2014. http://dx.doi. org/10.13140/rg.2.1.1494.2241 to our distinguished ladies we wish health, fruitful work and the huge satisfaction driver from it. more details in the scientific and academic activities of the celebrated ladies can be fund in the present issue. http://dx.doi.org/10.5586/mb.2004.001 http://dx.doi.org/10.5586/mb.2004.001 http://dx.doi.org/10.1007/978-94-011-2414-0 http://dx.doi.org/10.1007/978-94-011-2414-0 http://dx.doi.org/10.13140/rg.2.1.1494.2241 http://dx.doi.org/10.13140/rg.2.1.1494.2241 abstract acknowledgments references 2015-08-02t15:08:03+0100 piotr otręba endophytic fungi from vitex payos: identification and bioactivity 1 of 8published by polish botanical society acta mycologica original research paper endophytic fungi from vitex payos: identification and bioactivity edson panganayi sibanda1,2, musa mabandla2, tawanda chisango2, agness farai nhidza2, takafira mduluza2,3* 1 scientific and industrial research and development centre, food and biomedical technology institute, 1574 alpes road/scam way, harare, zimbabwe 2 school of laboratory medicine and medical sciences, college of health sciences university of kwazulu-natal, westville campus, private bag x54001, durban 4000, south africa 3 department of biochemistry, university of zimbabwe, po box mp167 mount pleasant, harare, zimbabwe * corresponding author. email: tmduluza@yahoo.com abstract endophytic fungi isolated from medicinal plants have an important role to play in the search for new bioactive natural compounds. however, despite their potential as repositories of bioactive compounds, the endophytes of african medicinal plants are largely underexplored. the aim of this study was to isolate and identify the endophytic fungi associated with vitex payos and evaluate their antimicrobial and antioxidant potential. the surface sterilization technique was used to isolate the endophytic fungi that were identified by rdna sequencing of the its region. crude methanol and ethyl acetate extracts were screened for antimicrobial activity using the agar diffusion method and evaluated for antioxidant activity using a commercial total antioxidant capacity assay kit. the total phenolic content of the extracts was determined using the folin–ciocalteu method and functional groups present in the extracts were predicted using fourier-transform infrared spectroscopy. seven endophytic fungi isolates identified as glomerella acutata, epicoccum nigrum, diaporthe species, penicillium chloroleucon, diaporthe endophytica, mucor circinelloides, and epicoccum nigrum were isolated from the tissues of vitex payos. none of the extracts exhibited antimicrobial activity and the crude ethyl acetate extract obtained from e. nigrum demonstrated both the highest total phenolic content (2.97 ±0.13 mg gae g−1 dry weight) and total antioxidant capacity (231.23 ±2.03 μm cre). fourier-transform infrared spectral analysis of the crude extracts from e. nigrum confirmed the presence of molecules carrying bonded hydroxyl functional group characteristic of phenolic compounds. these preliminary results indicate that most of the isolated fungal endophytes from v. payos belong to the phylum ascomycota and that the isolated e. nigrum strain has potential as a source of natural antioxidants. keywords diversity; antimicrobial; antioxidant; bioprospecting; africa; endophyte introduction there is heightened interest in bioprospecting for natural compounds with potential use as therapeutics owing to several factors that include the rapid development of antimicrobial resistant pathogenic microbes and emergence of new life-threatening diseases [1,2]. endophytes are microorganisms that live within plants for at least a part of their life cycle without causing any visible manifestation of disease [3]. endophytic fungi have been shown to produce a broad variety of bioactive secondary metabolites and bioprospecting of endophytes is considered a new frontier in the search for natural products with potential agricultural, pharmaceutical, and industrial applications [4,5]. doi: 10.5586/am.1111 publication history received: 2017-10-24 accepted: 2018-08-23 published: 2018-12-06 handling editor maria rudawska, institute of dendrology, polish academy of sciences, poland authors’ contributions tm and mm were responsible for the research design; eps, tc, and afn were responsible for data collection and analysis; tm and eps wrote the manuscript and mm revised the manuscript funding we are grateful to the university of kwazulu-natal (college of health sciences postgraduate research grant) for the financial support of this research. competing interests no competing interests have been declared. copyright notice © the author(s) 2018. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation sibanda ep, mabandla m, chisango t, nhidza af, mduluza t. endophytic fungi from vitex payos: identification and bioactivity. acta mycol. 2018;53(2):1111. https://doi. org/10.5586/am.1111 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:tmduluza%40yahoo.com?subject=endophytic%20fungi%20from%20vitex%20payos%3a%20identification%20and%20bioactivity https://doi.org/10.5586/am.1111 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1111 https://doi.org/10.5586/am.1111 2 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1111 sibanda et al. / endophytic fungi from vitex payos medicinal plants are rich sources of bioactive natural compounds and studies have shown that some medicinal properties of these plants may be related to the endophytic fungi that they host [2]. the endophytic fungi may participate in some of the plant metabolic pathways or may gain some genetic information to produce specific biologically active compounds, such as those produced by the host plant [6]. therefore, endophytic fungi isolated from medicinal plants have an important role to play in the search for new bioactive natural compounds. despite this potential of medicinal plants and the rich plant biodiversity in africa, only a tiny fraction of african medicinal plant species have been studied with regard to their endophytic fungi diversity. vitex payos is an african ethnomedicinal plant used to treat several ailments in zimbabwean traditional medicine [7]. as part of our contribution to the ongoing efforts to understand the diversity of endophytic fungi, we isolated and identified endophytic fungi found in the leaf and stem tissues of v. payos. furthermore, we determined the antibacterial activity and antioxidant potential for some of the identified endophytic fungi strains. in addition, fourier-transform infrared (ft-ir) spectroscopy was used predict the presence of various functional groups in the endophytic fungi crude extracts. material and methods collection and identification of plant materials fresh leaf and stem tissue of endophytic fungi were isolated from five leaf tissue samples and two stem tissue samples obtained from a single v. payos plant grown under natural (wild) conditions at the national herbarium and botanical gardens (harare, zimbabwe). the tissue samples were pretreated by cleaning them under running water to remove dirt and soil particles, and then air dried to remove any surface moisture before they were packaged into labeled sterile sample collecting bags. the samples were then transported to the laboratory and stored at 4°c until endophyte isolation procedures could be instituted [8]. isolation and establishment of in vitro culture of endophytes endophytic fungi were isolated from the leaf and stem tissues collected from the medicinal plant using a modified surface sterilization procedure as described by kjer et al. 2010 [9]. the plant tissue samples were removed from storage and thawed by washing them using running tap water. the thawed plant tissue samples were then washed using 0.1% (v/v) tween-80 for 15 minutes followed by another wash for 1.5 hours using running water. the cleaned plant tissues were then transferred to a laminar airflow cabinet where the surface sterilization and endophyte isolation was conducted under aseptic conditions. the plant tissue samples were cut into 5-cm segments and surface-sterilized with 70% ethanol (30 seconds for leaf samples and 2 minutes for stem samples), soaked in 2% naocl solution for 15 minutes, rinsed four times with sterile double distilled water, and finally blot-dried using sterile paper towels. the effectiveness of the sterilization procedure was tested by plating 0.1 ml of the final sterile water rinse onto petri dishes containing potato dextrose agar (pda) and rolling the sterilized sample onto petri dishes containing pda. the surface-sterilized tissues were cut into smaller segments (1–2 cm) using sterile razor blades and placed onto petri dishes containing pda supplemented with 200 u ml−1 penicillin-streptomycin (lonza, basel, switzerland; cat. no. 17-602e) and incubated at 28 ±2°c until fungal growth was initiated. the fungal mycelia growing out of the sample segments were subcultured and maintained on petri dishes containing pda. identification of the endophytic fungi genomic dna was extracted from the cultures using the zr fungal/bacterial dna kit (zymo research, irvine, ca, usa; cat. no. d6005). the internal transcribed 3 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1111 sibanda et al. / endophytic fungi from vitex payos spacer (its) target region was amplified using econotaq plus green 2x master mix (lucigen, madison, wi, usa) using the universal primers its1 and its4. the pcr reactions were carried out under the following conditions: initial denaturation at 95°c for 15 minutes, 35 cycles at 95°c (denaturation) for 1 minute, 56°c (annealing) for 30 seconds, 72°c (extension) for 1 minute, and a final extension for 10 minutes at 72°c. the pcr products were run on a gel and extracted (zymo research, irvine, ca, usa; zymoclean gel dna recovery kit; cat. no. d4001). the extracted fragments were sequenced in the forward and reverse directions using a bigdye terminator v3.1 cycle sequencing kit (applied biosystems, thermofisher scientific, waltham, ma, usa) and purified (zymo research, zr-96 dna sequencing clean-up kit; cat. no. d4050). the purified fragments were run on the abi 3500xl genetic analyzer (applied biosystems, thermofisher scientific). clc bio main workbench v7.6 was used to analyze the .ab1 files generated by the abi 3500xl genetic analyzer and endophytic fungi isolates were identified on the basis of similarity of amplified sequence with those found in the u. s. national center for biotechnology information (ncbi) database using the basic local alignment search tool (nblast). secondary metabolite extraction the small-scale fermentations and solvent extractions were carried as previously described [9] with modifications. briefly, the fungi were cultivated in 500 ml of potato dextrose broth and were incubated at 28 ±2°c for 30 days in a shaker at 180 rev min−1. the fungal mycelia were then separated from the culture broth by filtration and extracted with analytical grade methanol (meoh) solvent (100 ml). the filtrate was then extracted three times at the liquid-liquid partition with an equal volume of analytical grade ethyl acetate (etoac) solvent (1:1 v/v). the resulting crude extracts were collected and concentrated to dryness in a vacuum rotary evaporator at 40–45°c, then dissolved in 1 ml of meoh solvent, followed by drying under vacuum to obtain the etoac and meoh extracts. the crude extracts were dissolved in dimethyl sulfoxide (dmso) at a concentration of 1 mg ml−1 and kept at 4°c [10]. determination of the antimicrobial activity of the crude extracts the microorganisms were obtained from the american type culture collection (atcc, rockville, md, usa). the extracts were tested against the gram-positive bacteria staphylococcus aureus (atcc 11632), gram-negative bacteria escherichia coli (atcc1056) and klebsiella pneumoniae (atcc 13883) using the agar disk diffusion method using the clinical and laboratory standards institute protocol [11]. briefly, the test microorganisms were grown in nutrient broth medium until 1 × 108 colony-forming units were attained and then were used to inoculate sterile 90-mm petri dishes filled with nutrient agar medium using the spread-plate technique. dried and sterile filter paper disks (6.0 mm diameter) were impregnated with 40 µl of the extracts (containing 500 µg fungal extracts), air dried under aseptic conditions in a laminar airflow cabinet, and placed on plates inoculated with the test microorganism. the plates were incubated at 37°c for 24 hours. three sets of controls were used. the organism control consisted of a seeded petri dish with no sample. in the second control, samples were introduced to the unseeded petri dishes to check for sterility. disks impregnated with 40 µl dmso were run simultaneously as a third control. standard antibiotics were also run simultaneously as reference agents to understand the comparative antimicrobial efficacy. three replicates per extract were used for the antimicrobial activity assays. the antimicrobial potency of the extracts was measured by their ability to prevent the growth of the microorganisms surrounding the disks. determination of total phenolic content of the crude extracts the total phenolic content of the extracts was determined by the folin–ciocalteu method. briefly, 0.2 ml of each crude extract (dissolved in dmso at a concentration 4 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1111 sibanda et al. / endophytic fungi from vitex payos of 1 mg ml−1) was added to 2.8 ml of distilled water and mixed thoroughly with 0.5 ml of folin–ciocalteu reagent for 3 minutes, followed by the addition of 2 ml of 20% (w/v) sodium carbonate. the mixture was left to stand for a further 60 minutes in the dark and absorbance was measured at 650 nm using a model i-290 spectrophotometer (lasany, panchkula, india). the total phenolic content was calculated from the calibration curve, and the results were expressed as mg of gallic acid equivalent per g dry weight. three replicates per extract were used for the determination of total phenolic content and the mean values (±sd) were calculated. determination of the antioxidant activity of the crude extracts the total antioxidant capacity (tac) of the endophytic fungal extracts at 1 mg ml−1 concentration was determined using the oxiselect total antioxidant capacity assay kit (cell biolabs, inc., san diego, ca, usa) using the manufacturer’s instructions. the method is based on the single electron transfer (set) mechanism and involves the reduction of cu2+ to cu+ by the endogenous antioxidants and by other reducing equivalents in the sample. the cu+ interacts with a coupling chromogenic reagent that produces a color with a maximum absorbance at 490 nm. the absorbance value is proportional to the total antioxidant (respectively reducing) capacity of extracts. the samples were analyzed spectrophotometrically at 490 nm using the spectrostar nano microplate reader (bmg labtech, ortenberg, germany). tac was determined using a calibration curve based on uric acid standards. the results were expressed as μm copper reducing equivalents (cre). three replicates per extract were used for the determination of the antioxidant activity of the crude extracts and the mean values (±sd) were calculated. ft-ir analysis infrared spectra were collected on a nicolet 6700 ft-ir spectrometer (thermo fisher scientific) equipped with a diamond crystal attenuated total reflectance sampling accessory (thermo fisher scientific). a few grams of the samples were placed on the attenuated total reflectance unit and scanned from 500 to 4,000 cm−1 with a resolution of 4 cm−1. each recorded spectrum was the result of 36 coadded scans. ft-ir was performed to predict the presence of various functional groups in the isolates. data analysis the colonization rate of endophytes was determined as the total number of segments yielding ≥ 1 isolates divided by the total number of segments from which endophytes were isolated (expressed as a percentage). the results for the antimicrobial and antioxidant activity and total phenolic content determination assays are expressed as the mean ±sd of triplicates (n = 3). difference analysis (one-way anova) of the antioxidant activity results was conducted using microsoft excel 2013 (microsoft, usa). a p value <0.05 was considered to indicate statistically significant differences between groups. results isolation of endophytic fungi a total of seven endophytic strains were isolated from v. payos and the endophytic colonization rate was 100%. the isolated strains were identified (tab. 1) and all found to belong to the phylum ascomycota, except for mucor circinelloides, which is a member of the phylum zygomycota. 5 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1111 sibanda et al. / endophytic fungi from vitex payos antimicrobial activity none of the extracts exhibited antibacterial activity against the test microorganisms: e. coli (atcc1056), k. pneumoniae (atcc 13883), and s. aureus (atcc11632) as well as strains of the same bacteria obtained from clinical samples. antioxidant capacity and total phenolic content the antioxidant activity of etoac and meoh extracts of the two representative isolates obtained from stem (e. nigrum; isolate ts2) and leaf (diaporthe species; isolate tl3) tissues was determined. the etoac and meoh extracts of the endophytic fungi e. nigrum both exhibited antioxidant activity, while for the diaporthe species of endophytic fungi only the meoh extract showed some antioxidant activity (tab. 2). data analysis revealed significant differences [f(2, 6) = 8,429.77, p < 0.00001 at the 0.05 alpha level] in the antioxidant activities of the studied extracts, whereas there was no significant difference [f(1, 4) = 5.495, p = 0.07902 at the 0.05 alpha level] in the total phenolic content between the etoac extract from e. nigrum and the meoh extract from the diaporthe species. ft-ir analysis the etoac extracts from e. nigrum that exhibited both the highest antioxidant and total phenolic content were further analyzed using ft-ir spectroscopy, which is a well-established tool for the characterization and identification of functional groups present in extracts. the ft-ir spectral analysis of the etoac extract of e. nigrum revealed the presence of multiple functional groups, including –oh, –cho, –cooh, tab. 1 endophytic fungi isolated from the stems and leaves of v. payos. isolate code* sequence similarity with % sequence similarity accession number tl1 glomerella acutata 99 am991137 tl2 epicoccum nigrum 99 kx869952 tl3 diaporthe sp. 98 ku671340 tl4 penicillium chloroleucon 99 kp016813. tl5 diaporthe endophytica 96 ab899789 ts1 mucor circinelloides 100 kc461495 ts2 epicoccum nigrum 99 kx869952 * tl – endophyte isolated from leaf tissues; ts – endophyte isolated from stem tissues. tab. 2 total phenolic content and antioxidant capacity of extracts obtained from e. nigrum and diaporthe species*. endophyte isolate code total phenolic content (tpc) (mg gae/g extracta) total antioxidant capacity (tac) 1mg/ml [creb (μm)] ethyl acetate extracts epicoccum nigrum ts2 2.97 ±0.13 231.23 ±2.03 methanol extracts epicoccum nigrum ts2 0.50 ±0.01 32.28 ±2.96 diaporthe species tl3 2.76 ±0.07 60.97 ±2.53 * results are represented as means ±sd (n = 3). a gallic acid equivalent; b copper reducing equivalent. 6 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1111 sibanda et al. / endophytic fungi from vitex payos and –coor [12]. important ir absorption frequencies obtained from the extracts are tabulated in tab. 3. discussion the observation that most of the isolated endophytic fungi belonged to genera in the phylum ascomycota is in line with observations from other studies [13–15]. it is important to note that all the genera of endophytic fungi isolated from v. payos in this study have previously been isolated from a wide range of other different plant hosts in diverse environments, suggesting that these genera are not host and environment specific. the obtained preliminary antimicrobial activity results suggest that the studied endophytic fungi do not have potential as sources of new antimicrobial agents against the assessed bacterial species. while this is disappointing, the results support the findings from other studies that not all endophytic fungi strains have antibacterial activity [16]. however, there is a need to assess the antibacterial activity of these strains against other test microorganisms, screen them for antifungal activity, and vary the fermentation conditions before we totally discount these strains as potential sources of new antimicrobial agents. the etoac extract of the endophytic fungus e. nigrum exhibited both the highest antioxidant activity and total phenolic content. endophytic and marine-derived strains of e. nigrum have been shown to have antioxidant activity [17,18]. this suggests that this species might be generally predisposed to produce natural antioxidants. phenolic compounds have redox properties and are antioxidants owing to their hydroxyl groups that confer to them their free radical scavenging ability [15,19]. the total phenolic content and antioxidant activity results collected for the extracts obtained from the stem isolate of e. nigrum (ts2) suggest that phenolic content influenced antioxidant activity. however, there is a need for further studies to confirm this observation. endophytic species of the genus diaporthe have been reported to have antioxidant activity; our results for the meoh extract of our diaporthe leaf isolate tally with those observations. tab. 3 major bands observed in the ft-ir spectra of the e. nigrum etoac extracts. wave number (cm−1) vibration band/group probable compound classes 3,600–3,300 hydrogen-bonded o–h stretch phenols, alcohols 3,000–2,500 h–c–h asymmetric and symmetric stretch saturated aliphatic o–h stretch carboxylic acid s–h stretch thiols 1,820–1,680 c=o stretch carbonyls, lactones 1,600–1,550 c–c=c symmetric stretch aromatics –c=n– thiols and thio-substituted compounds c=o carbonyls n–h bend amines nitrogen-oxy aromatic nitro compounds 1,500–1,450 c–c=c asymmetric stretch aromatics 1,420–1,300 c=o carbonyls aliphatic nitro compounds hetero-oxy compounds dialkyl/aryl sulfones sulfur-oxy compounds 1,250–1,000 c–o stretch alcohols, phenols c–n aliphatic amines n–o aromatic amine oxide c=s thiocarbonyl φ–o–h aromatic ethers 7 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1111 sibanda et al. / endophytic fungi from vitex payos biological activity of extracts is influenced by the functional groups present in the extracts. hence, functional group analysis plays an important role in understanding the biological activity of extracts [20]. in the present study, the ft-ir spectral analysis of the e. nigrum crude etoac extracts suggests the presence of molecules carrying the bonded hydroxyl (–oh) functional group. the hydroxyl group is an integral part of most of the phenolic compounds, such as flavonoids and tannins. therefore, the ft-ir results serve to provide further supporting evidence for the presence of phenolic compounds in the crude etoac extracts. the ft-ir spectra analysis also suggests that diverse groups of functional groups (and hence diverse classes of compounds) are potentially present in the extracts. this is in line with previous findings by other researchers that endophytic fungi produce diverse classes of metabolites [21–23]. however, it is important to note that further investigations using different techniques are required to identify the compounds present in the e. nigrum crude etoac extracts. to the best of our knowledge, this is the first reported work on the identification, antibacterial and antioxidant activities of fungal endophytes isolated from v. payos. the results of this study suggest that while fungal endophyte isolates obtained from v. payos have potential as sources of natural antioxidants, further research is required to isolate and identify the bioactive molecules from the crude extracts and evaluate in vivo their biological activities. references 1. nguta jm, mbaria jm, gathumbi pk, gakuya d, kabasa jd, kiama sg. ethnodiagnostic skills of the digo community for malaria: a lead to traditional bioprospecting. front pharmacol. 2011;2:30. https://doi.org/10.3389/fphar.2011.00030 2. alvin a, miller ki, neilan ba. exploring the potential of endophytes from medicinal plants as sources of antimycobacterial compounds. microbiol res. 2014;169:483–495. https://doi.org/10.1016/j.micres.2013.12.009 3. stone jk, bacon cw, white jf. an overview of 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products: a review. microb pathog. 2015;82:50–59. https://doi.org/10.1016/j.micpath.2015.04.001 https://doi.org/10.1016/j.fbr.2007.05.003 https://doi.org/10.1002/mbo3.437 https://doi.org/10.1007/s00248-014-0379-4 https://doi.org/10.1016/j.micres.2009.11.009 https://doi.org/10.1007/s10126-011-9368-5 https://doi.org/10.1016/j.mrfmmm.2005.03.023 https://doi.org/10.1021/np058128n https://doi.org/10.3389/fmicb.2013.00065 https://doi.org/10.1016/j.micpath.2015.04.001 abstract introduction material and methods collection and identification of plant materials isolation and establishment of in vitro culture of endophytes identification of the endophytic fungi secondary metabolite extraction determination of the antimicrobial activity of the crude extracts determination of total phenolic content of the crude extracts determination of the antioxidant activity of the crude extracts ft-ir analysis data analysis results isolation of endophytic fungi antimicrobial activity antioxidant capacity and total phenolic content ft-ir analysis discussion references 2018-12-06t17:03:54+0000 piotr otręba bedrock and soil geochemistry influence the content of chemical elements in wild edible mushrooms (morchella group) from south italy (sicily) 1 of 12published by polish botanical society acta mycologica original research paper bedrock and soil geochemistry influence the content of chemical elements in wild edible mushrooms (morchella group) from south italy (sicily) maria grazia alaimo1, alessandro saitta2, elia ambrosio3* 1 department of earth and sea science, university of palermo, via archirafi 22, 90123 palermo, italy 2 department of agricultural, food and forest sciences, university of palermo, viale delle scienze, 90128 palermo, italy 3 via calamandrei 2, 53035 monteriggioni, siena, italy * corresponding author. email: elia.ambrosio.10@gmail.com abstract chemical elements in the samples of wild edible mushrooms of the morchella group collected from different unpolluted sicilian sites was analyzed by the icp-ms (method) to detect the content of their minerals and determine whether soil geology and geochemistry can influence the chemical composition in fungi. results showed that the mushroom samples mainly contained a high concentration of k and p and a wide variety of minor and trace elements (v, mo, pb, ce, cs, zr), including heavy metals. statistical analysis showed that the mushrooms differed in their content of minor and trace elements based on the geological/geographic site of origin. comparison with other studies showed differences in the content detected in the sicilian morels with those collected from other geographical sites. conversely, different fungal species collected from similar geological sites in sicily showed different patterns of accumulation of the elements confirming that bioconcentration in fungi is speciesand site-dependent. keywords fungi; icp-ms analysis; mineral content; accumulation factor; site geology introduction it is well known that fungi accumulate chemical elements from their environment, particularly from soils and soil solutions [1–6]. the mineralogical composition of soil influences the availability of chemical elements and the mycelia of fungi absorb and accumulate all kinds of elements from their growth substrates. although the capacity of accumulation and the presence of chemical elements in the fruiting bodies [7] of fungi depend on their nutritional requirements and can differ on the basis of genetic characteristics, several authors have confirmed that the content of elements in both microand macrofungi is mainly influenced by the chemical composition of the surrounding environment (water, air, and soil) [8–10]. the ability of fungi to take up elements makes them useful soil quality indicators as well as potential bioremediation agents for substrata contaminated with toxic elements such as heavy metals, metalloids, and radionuclides [11–16]. for instance, some authors [17] have observed that the macrofungal species agaricus macrosporus mont. may be effective in extracting heavy metals, such as mercury and cadmium, from contaminated soils. in several studies, the presence of heavy metals in edible and nonedible fungal species from both, polluted and unpolluted sites, has been analyzed [1–3,5,9,17]. however, till date, very few studies have actually analyzed the influence of geology, soil-mineralogy, doi: 10.5586/am.1122 publication history received: 2018-11-30 accepted: 2019-03-26 published: 2019-06-27 handling editor dorota hilszczańska, forest research institute, poland authors’ contributions mga and as set the field work and chemical analysis; ea made data analysis; all the authors contributed equally to the editing of the paper. funding this research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. publication fee has been paid by the research fund pj_ffabr_2017. competing interests no competing interests have been declared. copyright notice © the author(s) 2019. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation alaimo mg, saitta a, ambrosio e. bedrock and soil geochemistry influence the content of chemical elements in wild edible mushrooms (morchella group) from south italy (sicily). acta mycol. 2019;54(1):1122. https://doi. org/10.5586/am.1122 mailto:elia.ambrosio.10%40gmail.com?subject=bedrock%20and%20soil%20geochemistry%20influence%20the%20content%20of%20chemical%20elements%20in%20wild%20edible%20mushrooms%20%28morchella%20group%29%20from%20south%20italy%20%28sicily%29 https://doi.org/10.5586/am.1122 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1122 https://doi.org/10.5586/am.1122 2 of 12© the author(s) 2019 published by polish botanical society acta mycol 54(1):1122 alaimo et al. / chemical elements in morels and soil-chemistry on the chemical content of the fruiting bodies or investigated the correlation between chemical elements in fungi and their soil of growth [4,8,11,18]. for instance, nikkarinen and mertanen [8] analyzed the content of elements in two ectomycorrhizal species namely boletus edulis bull. and lactarius trivialis (fr.) fr. from two different geological regions in finland to determine whether any geochemical fingerprints can be observed in these fungi. they found that the macrofungal samples differed considerably in their content of trace elements based on the geological and geographic site of origin. nonnis marzano et al. [18] used chemical and radiochemical methodologies to analyze the concentrations of artificial radionuclides and trace elements in boletus samples (known as “porcini”) collected from different geographical areas across the globe. their results showed that the content of chemical and radiochemical elements in the fruiting bodies reflect the geological/chemical background of the environment and, therefore, can be used to determine the geographic site of origin. analyzing the mineral content and correlating it with the bedrock and soil geochemistry is particularly important for wild edible mushrooms due to their economic and social importance. over 2,000 fungal species are known to produce edible fruiting bodies that are harvested, marketed and consumed in more than 85 countries around the world [19–23]. the global market value of edible mushrooms is estimated to be at least $2 billion, which is more than the value of timber [19,24,25]. when toxic elements (e.g., heavy metals) are present in the growth substrates, they can accumulate in the fungi and may pose a risk to human health. particularly, the content of metals and metalloids is likely to be higher in mushrooms than those in agricultural crops, plants, vegetables, and fruits [6]. in this paper, the following approaches were undertaken to determine whether the bedrock and soil chemistry influences the content of elements in wild mushrooms: (i) we analyzed the chemical content in the wild edible fruiting bodies of morels (morchella spp.) collected from different unpolluted sites in the south of italy (sicily). our objective was to compare the content of major, minor, and trace elements in the fruiting bodies of mushrooms obtained from different geological sites in sicily and investigate whether the observed differences could be correlated to those in the bedrock and soil geochemistry; (ii) moreover, using a set of literature data on the bedrock and (top) soil composition in sicily, italy, europe and the rest of the world, we determined the accumulation factor (a.f.) of different elements in the fungal samples to assess which elements are accumulated in mushrooms at concentrations higher than those in the soil; (iii) finally, we compared the data thus obtained with those of mushrooms analyzed from other geographic/geological sites to determine whether soil geology and geochemistry can influence the chemical components present in the fungi; (iv) we also compared our data with those of other fungal species to understand whether different species of edible wild mushrooms originating from similar geological sites of sicily differ in their preferences for the accumulation of specific elements. two species within the morchella genus, m. elata fr. and m. esculenta (l.) pers., were chosen for this study because they are the ones that are majorly consumed globally [22] and give an estimated worldwide income of ~$1.67 billion [19,26,27]. material and methods study sites ten sampling sites were selected from the province of palermo in sicily between the natural regional park of madonie and the palermo mountains (fig. 1). tab. 1 gives complete details on the study sites. overall, we selected different mediterranean vegetation types including both native (e.g., broadleaf and evergreen plants) and planted (e.g., conifers) wooded areas. the geology of the selected sites consisted of dolomitic limestone, carbonate rocks (s1–s3 and s6–s10), and flyschoid rocks (s4–s5). the climate in the province of palermo is the mediterranean pluviseasonal-oceanic type [28]. the mean annual temperature is 18.4°c and varies from 26.2°c in august to 12.1°c in january. the mean annual rainfall is 605 mm and varies from 90 mm in december to 4 mm in july [29]. 3 of 12© the author(s) 2019 published by polish botanical society acta mycol 54(1):1122 alaimo et al. / chemical elements in morels fig. 1 map of the study sites and the picture of m. esculenta. 4 of 12© the author(s) 2019 published by polish botanical society acta mycol 54(1):1122 alaimo et al. / chemical elements in morels sampling during the season favorable for fungal growth, in april and may 2014, several mycological surveys were carried out at the selected study sites. the fruiting bodies/mushrooms, namely m. elata fr. and m. esculenta (l.) pers. (f1–f10 in tab. 1), were sampled from 10 unpolluted sites of sicily (refer fig. 1 and tab. 1). each sample consisted of 1–3 fungal specimens comprising a complete fruiting body (cap, stipe, and hymenium). samples of the mushrooms were cleaned of forest debris (without washing) using a brush, transported to the laboratory and kept at −4°c for no more than 24 h prior to sample preparation. identification of the species and description of the fruiting bodies/ascomata were carried out for both fresh and dried specimens by macroand microscopic observations. the macroscopic descriptions of the fresh ascomata were noted while the microscopic features were observed using an olympus bh-2. the dried specimens were prepared for microscopic observations using a solution of 0.3% koh and the melzer reagent. spore measurements were based on 50 observations conducted for each of the dried samples. for taxonomical identification, a series of monographs and keys were used by breitenbach and kränzlin [30], courtecuisse r and duhem [31], and boccardo et al. [32]. we followed the methods of systematic classification described previously for the classification of mushrooms [7,33]. nomenclature and author abbreviations were used in accordance with [34–36]. studied specimens were deposited at the herbarium panormitanus (pal), italy. tab. 1 details of the study sites: code, locality with gps coordinates, geology, and vegetation type. fungal species and their corresponding codes are detailed in the last two columns. site locality – municipality geology vegetation type fungal species mushrooms code s1 monte petroso – monreale (38°06'03.1" n, 13°15'43.2" e) carbonate rocks quercus ilex – native wood morchella elata f1 s2 castellaccio – monreale (38°04'49.5" n, 13°15'48.9" e) carbonate rocks pinus pinea – plantation m. elata f2 s3 castellaccio – monreale (38°05'05.4" n, 13°15'58.2" e) carbonate rocks pinus pinea – plantation m. elata f3 s4 fontana bosco – palermo (37°53'34.8" n, 13°23'28.4" e) flyschoid rocks quercus cerris var. gussonei and fraxinus angustifolia – native mixed wood m. esculenta f4 s5 fontana bosco – palermo (37°53'21.7" n, 13°23'33.4" e) flyschoid rocks quercus cerris var. gussonei and fraxinus angustifolia – native mixed wood m. esculenta f5 s6 mandria zarcati – carini (38°09'12.5" n, 13°16'04.3" e) carbonate roks pinus halepensis – plantation m. elata f6 s7 mandria zarcati – carini (38°09'08.8" n, 13°15'52.9" e) carbonate rocks pinus halepensis – plantation m. elata f7 s8 pizzo colla – polizzi generosa (38°09'08.8" n, 13°15'52.9" e) carbonate rocks fagus sylvatica – native forest m. elata f8 s9 bevaio del faggio – isnello (37°52'14.6" n, 14°00'33.4" e) carbonate rocks pinus nigra – plantation m. elata f9 s10 bevaio del faggio – isnello (37°52'14.6" n, 14°00'33.4" e) carbonate rocks fagus sylvatica – native forest m. elata f10 5 of 12© the author(s) 2019 published by polish botanical society acta mycol 54(1):1122 alaimo et al. / chemical elements in morels chemical analysis the fruiting bodies were dried for 12 hours at 37°c in an electrically heated commercial dehydrator for mushrooms, fruits, and vegetables (melchioni 118320000 babele, 245 w). the dried fungal material was ground into powder using an agate mortar and stored in polyethylene bags under dry conditions. powdered mushrooms (~0.700 g) were digested using a mixture of 5 ml of 65% hno3 (suprapure, merck) and 2.5 ml of 33% h2o2 (suprapure, merck). the digest was diluted to 50 ml using deionized water (18 mω). ca, k, na, and mg were analyzed using the ion chromatograph dionex 120, with a precision greater than ±5%. the presence of 24 elements (ag, al, as, ba, be, bi, cd, co, cr, cu, fe, li, mn, mo, ni, pb, rb, sb, se, sr, tl, u, v, and zn) in the digested extract was determined using an inductively coupled plasma-mass spectrometer (icpms) (elan 6100 drc-e, perkinelmer). for the detection of as, cr, fe, se, and v, the icp-ms was operated in the drc (dinamic reaction cell) mode with methane as the reaction gas. all standard solutions were prepared with ultra-pure deionized water (18 mω) and reagent-grade chemicals (icp multielement standard solution xxi certipur – merck; mo and sb, certipur standards – merck). calibration curves ranging from 0.05 μg/l to 500 μg/l were constructed. the standard addition technique was used for all analyses in order to minimize the matrix effects. sample blanks were also analyzed and the operational detection limit for each element was calculated as three times the standard deviation of the analyte concentration in the blank samples. values below the detection limit were set at one-third of the detection level and treated as real values. analytical precision was in the range 1–11% for all the analyzed elements. for validation of the analytical procedure, the standard reference material nist srm 1515 apple leaves was analyzed for the corresponding elements. the rates of recoveries for metal were in good agreement with the certified concentrations, ranging between 94% and 111%. all the analyzed elements are detailed as follows: ■ alkali metals: li, na, k, rb, cs. ■ alkaline earth metals: mg, ca, sr, ba. ■ transition metals or d-block elements: y, ti, zr, hf, v, nb, ta, cr, mo, w, mn, re, fe, co, ni, pd, pt, cu, ag, au, zn, cd, hg. ■ semimetals: b, ge, as, te, sb. ■ post-transition metals: al, ga, tl, sn, pb, in, bi. ■ lanthanides: la, ce, pr, nd, sm, eu, gd, tb, dy, ho, er, tm, yb, lu. ■ actinides: th, u. ■ nonmetals: p, se. in total, the concentrations of 63 elements were determined (see appendix s1 for a complete list) and converted from ppb to ppm for statistical analyses, as detailed in the following data analysis paragraph. data analysis quantitative data, referred to as the chemical concentrations of the elements in ppm, were analyzed statistically using the vegan package in r [37]. descriptive statistical tools (histograms) were used to compare the concentrations of major, minor, and trace elements in each of the fungal samples. we also used the whole dataset (see appendix s1) to detect the a.f. in the fungal samples. the a.f. refers to the ratio of the concentration of a specific element in the mushroom samples to the concentration of that element in the soil (or native rock/ bedrock). the a.f. was computed in accordance with cocchi et al. [9] by comparing the concentration of each element detected in the fungal samples with the average concentrations of the same element in the soil. as a proxy for the average concentrations of elements in the soil, we used known concentrations of the elements in comparable soil types (appendix s2). this information was obtained from a set of literature data on the bedrock and (top) soil composition in sicily, italy, europe and the rest of the world [38–49]. histograms were used to indicate the accumulation factor. 6 of 12© the author(s) 2019 published by polish botanical society acta mycol 54(1):1122 alaimo et al. / chemical elements in morels finally, a hierarchical cluster analysis (ca) using the bray–curtis dissimilarity index and unweighted pair group method with arithmetic mean upgma [50] were carried out to discern the degree of similarity between the content of chemicals in the mushroom samples and the selected bedrocks or soil types. results appendix s1 lists all the 63 chemical elements detected in each of the mushroom samples (f1–f10). the values of major elements (20 in total) measured in the mushroom samples ranged from 0.01 ppm (sb) to 4,300 ppm (k) (fig. 2) in the following order of abundance: k > p > ca > mg > na > a > fe > zn > cu > mn > rb > ti > cd > au > sr > b > ba > cr > ni > sb. minor and trace elements (26 in total) ranged from 0.001 ppm (u) to 1.26 ppm (v) (fig. 3) in the following order of abundance: v > mo > pb > ce > cs > zr > ag > li > se > y > co > la > as > nd > ga > tl > hg > nb > th > pr > sm > gd > ge > yb > te > u. seventeen other elements were detected under the limits of this detection method (see appendix s1). overall, the concentrations of the major elements showed a very similar distribution across the mushroom samples (fig. 2); however, in the cases of minor and traces elements (fig. 3), different patterns were observed in the collected mushrooms. for instance, in f3, we found a higher concentration of pb, v, zr, and ce than that in the other samples. when comparing the concentrations of elements in the mushroom samples (f1–f10, see appendix s1) with those in the carbonate rocks, according to the selected literature datasets, flyschoid rocks + clays, earth’s crust and topsoil of sicily, italy, europe, and the rest of the world (appendix s2), we could not conclude whether some elements 0 5,000 10,000 15,000 20,000 25,000 30,000 35,000 40,000 45,000 50,000 al au b ba ca cd cr cu fe k mg mn na ni p rb sb sr ti zn pp m f 1 f 2 f 3 f 4 f 5 f 6 f 7 f 8 f 9 f 10 fig. 2 major elements content in mushroom samples (f1–f10). 7 of 12© the author(s) 2019 published by polish botanical society acta mycol 54(1):1122 alaimo et al. / chemical elements in morels are accumulated better than the others, because the concentrations of such elements in the analyzed mushroom samples were below the detection limits. some other elements show an a.f. that can be clearly correlated to the average value of those elements detected in their corresponding bedrocks and topsoil (fig. 4). particularly, an accumulation of the following six elements was detected in the sampled mushrooms: ag, cd, cu, k, p, and zn (see fig. 4). appendix s3 represents the a.f. for each mushroom sample and the reference soil type. results of the ca in fig. 5 show that the samples of morels (f1–f10) and the rock/ soil types used for comparison (carbonate rocks, earth’s crust and topsoil from sicily, italy, europe, and the rest of the world) form two distinctive clusters and the mushroom samples are placed into clusters on the basis of the geographic and geologic site of collection. discussion based on the results on samples of morchella spp. obtained in this study, we can confirm that fungi can accumulate different elements from their substrata (especially soil) of the areas where they grow. the content of elements in the morels used for this study consisted of high concentrations of k and p and a wide pattern of minor and trace elements (e.g., v, mo, pb, ce, cs, zr), including heavy metals. moreover, when the content of elements in the studied specimens was compared with the average concentration values in the bedrock and soil types (see appendix s2), it was observed that the morels had accumulated six elements in particular which included the ones found abundantly in the earth’s crust (k, p, and zn; fig. 4) and heavy metals such as ag, cd, and cu. these results confirm that fungi are characterized by the presence of high concentrations of k in their structures, which are comparable with those found in some agricultural crops, plants, and vegetables such as spinach and potatoes [6], as well as the presence of (heavy) metals like cd and pb. it is worth noting that the content of cd in the soil is mainly influenced by the native rock (e.g., high in carbonate rocks) and is particularly abundant in anthropized soil. moreover, the concentrations of cd and pb in the analyzed mushrooms were below the threshold limits for wild mushrooms specified by the european directives [51,52]. f 1 f 2 f 3 f 4 f 5 f 6 f 7 f 8 f 9 f 10 ag 0.2 0.056 0.122 0.2 0.056 0.289 0.233 0.222 0.289 0.367 as 0.165 0.153 0.225 0.093 0.075 0.06 0.06 0.072 0.101 0.147 ce 0.441 0.358 0.843 0.188 0.161 0.03 0.04 0.162 0.025 0.093 co 0.252 0.164 0.277 0.114 0.062 0.061 0.085 0.102 0.091 0.104 cs 0.451 0.362 0.346 0.0478 0.102 0.147 0.182 0.063 0.0497 0.476 ga 0.164 0.139 0.236 0.01 0.024 0.04 0.041 0.047 0.022 0.047 gd 0.045 0.035 0.081 0.009 0.008 0.002 0.003 0.014 0.002 0.007 ge 0.02 0.03 0.02 0.01 0.01 0.01 0.01 0.02 0.01 0.02 hg 0.02 0.06 0.05 0.03 0.02 0.04 0.03 0.04 0.04 0.06 la 0.266 0.196 0.488 0.099 0.086 0.016 0.02 0.079 0.015 0.047 li 0.41 0.33 0.67 0.1 0.07 0.09 0.07 0.12 0.03 0.12 mo 0.416 0.452 0.343 0.224 0.23 0.247 0.199 0.369 0.233 0.284 nb 0.065 0.043 0.125 0.023 0.012 0.005 0.005 0.017 0.023 0.01 nd 0.235 0.184 0.446 0.072 0.061 0.013 0.02 0.07 0.011 0.039 pb 0.43 0.53 1.09 0,0 2 0.05 0.05 0.06 0.1 0.02 0.08 pr 0.059 0.046 0.109 0.021 0.019 0.005 0.005 0.018 0.005 0.01 se 0.03 0.24 0.21 0.03 0.03 0.1 0.15 0.27 0.49 0.25 sm 0.048 0.036 0.082 0.013 0.01 0.004 0.005 0.014 0.002 0.009 te 0.014 0.011 0.013 0.003 0.003 0.004 0.008 0.003 0.003 0.003 th 0.06 0.05 0.11 0.02 0.02 0.01 0.01 0.02 0.01 0.01 tl 0.132 0.129 0.093 0.004 0.005 0.039 0.052 0.016 0.002 0.078 u 0.016 0.011 0.018 0.003 0.003 0.022 0.001 0.004 0.001 0.004 v 0.8 0.62 1.26 0.15 0.16 0.08 0.06 0.24 0.07 0.31 y 0.383 0.22 0.549 0.025 0.025 0.013 0.017 0.052 0.009 0.043 yb 0.026 0.016 0.041 0.003 0.002 0.002 0.001 0.003 0.001 0.004 zr 0.5 0.3 0.89 0.14 0.07 0.02 0.03 0.07 0.02 0.06 0 0.2 0.4 0.6 0.8 1 1.2 1.4 ag as ce co cs ga gd ge hg la li mo nb nd pb pr se sm te th tl u v y yb zr pp m f 1 f 2 f 3 f 4 f 5 f 6 f 7 f 8 f 9 f 10 fig. 3 minor and trace elements in the mushroom samples (f1–f10). elements very close or under the detection limit were not included in this graph. 8 of 12© the author(s) 2019 published by polish botanical society acta mycol 54(1):1122 alaimo et al. / chemical elements in morels fig. 5 cluster dendrogram of matrix distance of the mushroom samples and soil types (carbonate rocks, earth’s crust and topsoil from sicily, italy, europe, and the rest of the world) based on the content of their elements. fig. 4 content of ag, cd, cu, k, p, and zn in mushroom samples (f1–10) and the rocks and soil types. 0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 pp m ag 0 5 10 15 20 25 pp m cd 0 10 20 30 40 50 60 70 pp m cu 0 5,000 10,000 15,000 20,000 25,000 30,000 35,000 40,000 45,000 50,000 pp m k 0 5,000 10,000 15,000 20,000 25,000 pp m p 0 50 100 150 200 250 300 pp m zn 9 of 12© the author(s) 2019 published by polish botanical society acta mycol 54(1):1122 alaimo et al. / chemical elements in morels statistical analysis showed that the mushroom samples (f1–f10) were clustered on the basis of the soil type. samples f1–f3 and f6–10 (except f9) were collected from the carbonate rock and formed a separate group (viz. sister group) from samples f4– f5, which were collected from sites with a substratum developed on flyschoid rocks. similarly, the second cluster (see right side in fig. 5) was established by bedrock and soil types (e.g., topsoil, earth’s crust, carbonate and flyschoid rocks) and samples therein formed sister groups on the basis of their geological differences. based on these results, we can observe that both site geology and geochemistry can influence the chemical composition in wild mushrooms. it is challenging to compare our results with those of others because, till date, very few studies are available that have analyzed all the elements in morels and evaluated the influence of bedrock or soil geochemistry in this fungal group. for instance, cenci et al. [53] studied heavy metals in samples of some morels collected from central italy (in the emilia-romagna region) and found a higher content of hg (0.28–2.7 mg/kg) than that observed in the sicilian morels of our study (hg = 0.05–0.33 mg/kg). conversely, the content of cd detected in our samples (0.18–21.5 mg/kg) was higher than that in the morels of central italy (cd = 0.19–4.12 mg/kg [53]), or in some morchella species found in turkey (cd = 0.036–1.43 mg/kg [54]). despite a differences in the geological sites, the cu content in our samples (10.6–63 mg/kg) was very similar to that detected from the morels of central italy (43.65–63.39 mg/kg), whereas the content of ag in the sicilian samples (0.05–0.22 mg/kg) was lower than that found by cocchi et al. [9] (ag = 0.28–2.7 mg/kg). we were also interested in understanding whether different species of edible wild mushrooms collected from similar geological sites would exhibit differences in the accumulation of elements. we compared our dataset with that of venturella et al. [4] and alaimo et al. [6], including information on the mineral contents of some bolets [e.g., boletus aereus bull., b. reticulates schaff., b. impolitus fr., b. lupinus fr., b. queletii, b. rhodoxanthus (krombh.) kallenb., b. satanas lenz, and leccinum lepidum (h. bouchet ex essette) bresinsky & manfr. binder] and clitopilus prunulus p. kumm collected in the sedimentary sites (flyschoid or calcareous substrates) of sicily. although all species had a high concentration of k and na [4,6], the bioconcentration of the other elements appeared to be elementand species-depended. for example, co, cr, fe, mg, mo, pb, u, and v was four folds higher in c. prunulus than that detected in the bolets analyzed from similar geological sites. to summarize, the results obtained in this study confirmed that fungi (here, the morchella group) accumulate all kinds of elements (including heavy metals) and that the bedrock or soil geochemistry can influence patterns of their minerals. the chemical content of elements in our morel samples was characterized by high concentrations of k and p. additionally, a wide number of minor and trace elements, including heavy metals, were accumulated in the fruiting bodies. a comparison of our results with those of other studies demonstrated that the same fungal species (in this case, morchella spp.), collected from different geological and geographic sites was characterized by a differing content of minor and trace elements in their bodies. conversely, different fungal species collected in similar geological sites (in sicily) exhibited different patterns of accumulation of the elements. this confirms that, in fungi, the bioconcentrations are species-dependent and site-dependent. in fact, different fungal species have shown different responses to metal pollution. on the other hand, the content of elements in the soil depends on both physical characteristics (e.g., the grain of soil) and the impact human activities [55,56]. future studies should include samples from different geographical/geological sites and other species of edible mushrooms. with growth in the consumption and demand of edible wild mushrooms, the analysis of mineral content in both mushrooms and soils can be an important tool in identifying the possible risks to human health. 10 of 12© the author(s) 2019 published by polish botanical society acta mycol 54(1):1122 alaimo et al. / chemical elements in morels acknowledgments authors are gratefull to v. agamennone and to the managing/production editor for linguistic revision; g. nardi for the graphic images. anonymous reviewers are also acknowledged for useful comments and observations. supplementary material the following supplementary material for this article is available at http://pbsociety.org.pl/ journals/index.php/am/rt/suppfiles/am.1122/0: appendix s1 list of elements detected in each mushroom sample (f1–f10). appendix s2 element content in carbonate rocks, earth’s crust and topsoil from sicily, italy, europe, and the rest of the world. appendix s3 list of the mushroom samples (f1–f10) with the accumulation factor based on the types of bedrocks and soils (soil and rock from sicily, italy, europe, and the rest of the world) were considered. references 1. demirbas a. accumulation of heavy metals in some edible mushrooms from turkey. food chem. 2000;68:415–419. https://doi.org/10.1016/s0308-8146(99)00210-1 2. svoboda l, havličková b, kalač p. contents of cadmium, mercury and lead in edible mushrooms growing in a historical silver-mining area. food chem. 2006;96:580–585. https://doi.org/10.1016/j.foodchem.2005.03.012 3. yamaç m, yildiz d, sarikürkcü c, çelikkollu m, solak mh. heavy metals in some edible mushrooms from the central anatolia, turkey. food chem. 2007;103(2):263–267. https://doi.org/10.1016/j.foodchem.2006.07.041 4. venturella g, gargano ml, compagno r, saitta a, alaimo mg. the mineral contents of some boletaceae species from sicily (southern italy). j aoac int. 2014;97(2):612–623. https://doi.org/10.5740/jaoacint.12-260 5. zhang d, gao t, ma p, luo y, su p. bioaccumulation of heavy metal in wild growing mushrooms from liangshan yi nationality autonomous prefecture, china. wuhan university journal of natural sciences. 2008;13(3):267–272. https://doi.org/10.1007/s11859-008-0302-2 6. alaimo mg, dongarrà g, la rosa a, tamburo e, vasquez g, varrica d. major and traces elements in boletus aereus and clitopilus prunulus growing on volcanic and sedimentary soils of sicily (italy). ecotoxicol environ saf. 2018;157(15):182–190. https://doi.org/10.1016/j.ecoenv.2018.03.080 7. kirk pm, cannon pf, minter dw, stalpers ja. dictionary of the fungi. 10th ed. wallingford: cab international; 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e-mail: manal.adnani@gmail.com abstract fungal isolates of fusarium were collected from symptomatic chickpea (cicer arietinum l.) plants growing in fields within souk tlat commune in the gharb region. morphological and molecular characterizations were performed of the fungal isolate n3 obtained from a chickpea plant. pcr amplification and sequencing of the internal transcribed spacer using the primers its1 and its4 was applied to identify the fungal isolate n3. the maximum similarity index of the fungus was found to be 99.33% with fusarium equiseti (accession no. mt111122). in the pathogenicity test, both chickpea seed dip inoculation and soil infestation by the spore suspension of fusarium isolate were adopted. four weeks after chickpea seed inoculation, few plants emerged and those that emerged were stunted. a high percentage of inoculated seeds did not emerge and po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 1 mailto:manal.adnani@gmail.com showed accentuated rot symptoms.. eight weeks after sowing seeds in infested soil, the obtained chickpea seedlings displayed root necrosis, browning at the crown, and wilting. in addition, these plants showed a foliar alteration index of 0.395. the re-isolation was positive for different parts of chickpea plants for both seed and soil inoculation. fusarium equiseti isolate decreased the length of the root and aerial parts, and number of leaves and branches of the inoculated chickpea plants either by seed inoculation or soil infestation with values of 0.91 cm and 19.73 cm, 1.29 cm and 19.44 cm, 1.11 and 18.66, and 0.0 and 2.08 respectively, whereas the corresponding values for the control plants were 27.16 and 28.33 cm, 29.05 and 31.05 cm, 24.21 and 25.66, and 3.50 and 3.11, respectively. to the best of our knowledge, this is the first report of f. equiseti on chickpea (cicer arietinum l.) in morocco. keywords – chickpeas – inoculation – fusarium equiseti – rot – wilting running head: fusarium equiseti in chickpea: first report in morocco 1. introduction chickpea (cicer arietinum l.) is one of the oldest cultivated protein legumes in the world. it is mainly used for human consumption and is an essential constituent of the mediterranean diet and basic food in pakistan and india (millan et al., 2010). domesticated in association with other crops such as wheat and barley, cicer arietinum l. is believed to be a part of the agricultural revolution, and in terms of consumption, ranks second after broad bean (gupta et al., 2014). in morocco, the cultivated chickpea area covers an acreage of 54000 ha (mapmdref, 2020a), with a production of 49700 t recorded in the 2019/2020 agricultural season (mapmdref, 2020b). most of the agricultural systems of chickpea production in the country suffer from several constraints, mainly biotic and abiotic factors that cause serious damage before and after harvest. the widespread high temperature and drought stress in different regions of chickpea production po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 2 can affect flowering and pod setting stages which lead to decreases in chickpea yield (houasli et al., 2020). previous reports on the chickpea crop have recorded the presence of ascochyta rabiei, the causal agent of blight disease in morocco, in all chickpea areas (grewal, 1984; singh, 1984)bencheqroun et al. (2022) pointed out that didymella rabiei (kovatsch.) arx. is the most devastating fungal infection of chickpea crops inflicting considerable yield and quality losses. moreover, numerous fungal species associated with the chickpea diseases of wilt and root rot have been reported in morocco, including fusarium oxysporum f sp. ciceris (el aoufir, 2001; el bouazaoui et al., 2018); fusarium redolens (jiménez-fernández et al., 2011); and rhizoctonia bataticola, r. solani, and pythium sp. (el bouazaoui et al., 2018). fusarium equiseti has also been reported in other crop species, especially melon (cucumis melo), soybean (glycine max), cumin (cuminum cyminum), cauliflower (brassica oleracea), winter rapeseed (brassica napus), tomato plant (solanum lycopersicum), pepper (capsicum annuum), and the cabbage brassica oleracea (adams et al., 1987; chen et al., 2014; gally et al., 1998; goswami et al., 2008; li et al., 2014; ramchandra & bhat 2012). the pathogen is also responsible for pre and postharvest decay of zucchini fruits (cucurbita pepo l.) (ezrari et al., 2019). it was recently isolated from the seeds of the fragrant wallflower (matthiola longipetala) showing rot (ivanović et al., 2020). khan et al. (2021) noted that f. equiseti was responsible for seedling death in sugar beet. isolates of fusarium and f. solani were recovered from necrotic lesions of chickpea roots in different chickpea growing areas (el hazzat et al., 2019). nevertheless, taxonomic confirmation of which species of fusarium causes this necrosis is lacking because numerous species are important plant pathogens (austwick, 1982). additionally, the differentiation of fusarium species po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 3 through morphological characters is imprecise; hence, the use of molecular techniques become more efficient and accurate for the discrimination of fungal species (steenkamp et al., 2000). therefore, the present study was carried out to identify an isolate of fusarium sp. collected for the first time in chickpea fields in morocco from diseased chickpea plants based on morphological characters and molecular and pathological characterization via fulfillment of koch’s postulates. 2. materials and methods 2.1 fungal material fusarium isolates was obtained from necrotic lesions associated with infected stem samples of chickpea plants that were grown in different fields in souk tlat in gharb province of the rabatsale-kenitra region, morocco. one hundred plants were chosen at random from the fields. one stem base sample from each plant was analyzed. pieces of diseased tissues were rapidly disinfected with 90% alcohol for 5 min, rinsed three times with sterile distilled water, and dried with sterile filter paper. samples were then placed onto potato dextrose agar plates (biokar diagnostics) and incubated at 25 °c for 7 days. the colonies formed were transferred to potato sucrose agar (psa) medium containing potato, sucrose, agaragar, and distilled water. agar plates were incubated in the same conditions and then observed for species determination. the fusarium sp. isolate was cultivated in petri dishes containing psa medium. the medium was poured into petri dishes containing 100 mg/l of chloramphenicol at a rate of 30 to 40 ml per dish. incubation of cultures was performed in the dark at 25 °c for 7 days followed by macroscopic and microscopic characterization depending on the age of the cultures. 2.2 morphological characterization macroscopic examination of fusarium sp. was carried out according to the development of the cultures on psa medium. observations focused on colony appearance, mycelium density, the po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 4 presence of the pinkish color of the colony, as well as growth and spore production. the microscopic characteristics of the fusarium n3 isolate were determined under an optical microscopic to confirm the species identity of this pathogen. 2.3 molecular analysis and identification molecular identification of the fusarium n3 isolate was performed after 5 days of culture on psa medium. dna extraction was performed according to the method described by murray and thompson (1980) and doyle et al. (1987). dna amplification of the internal transcribed spacer (its) rdna region was performed using polymerase chain reaction (pcr) using universal primers its1 (5′-tccgtaggtgaacctgcgg-3′) and its4 (5′-tcctccgcttattgatatgc-3′) (white et al., 1990). the pcr reaction was carried out in a reaction mixture of 25 μl containing 5 µl of 5× buffer (mytaq reaction buffer, bioline, london, uk,), 1 µl of each primer (10 µm), 0.2 µl of mytaq dna polymerase (bioline, london, uk) (5 u µl-1), 1–2 μl of template dna (100 ng) and milli-q water to complete the volume. a veriti thermal cycler (applied biosystems) was used for the pcr with the following conditions: initial denaturation at 95 °c for 1 min; 35 cycles of denaturation at 95 °c for 15 s, annealing at 52 °c for 20 s, and extension at 72 °c for 15 s; and a final elongation of 72 °c for 3 min. the quality of the pcr products was verified by electrophoresis on 1% agarose gel in the presence of a 100 bp molecular weight marker. sequencing was performed using an abi prism bigdye terminator v.3.1 ready reaction cycle sequencing kit and primer set its1 and its4. the sequencing products were run on an abi prism 3130xl genetic analyzer (applied biosystems) using the pop-7 polymer. the sequence resulting from this study was submitted to genbank under the accession no. mt111122, the obtained its sequence was then compared with the homologous nucleotides sequence in the genbank database using the basic local alignment search tool (blast) po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 5 (http://www.ncbi.nlm.nih.gov/blast). the seeds of a local variety of chickpea intended for pathogenicity testing were surface sterilized by soaking for 5 min in a 10% naocl solution, rinsed in sterile distilled water, and then dried on filter paper. the pathogenicity of the n3 isolate was checked through two inoculation techniques. technique 1: the surface-disinfected chickpea seeds were inoculated by soaking in water containing a conidial suspension of 106 spores/ml of the fusarium isolate n3 at room temperature (20 °c) for 1 h. the control seeds were treated with only sterile distilled water. the inoculated and control chickpea seeds were sown in plastic pots (13 cm × 13.5 cm) containing autoclaved sieved mamora forest soil at the rate of 5 seeds/pot. the mamora forest soil used is loose, very sandy, and slightly basic ph (7.27), with an organic carbon content of 0.35% to 0.6% (mouria, 2009). the soil was sieved and sterilized three times at an interval of 24 h at 200 °c for 2 h, and then distributed in the plastic pots (13 cm × 13.5 cm) at the rate of 2 kg of soil per pot. technique 2: the culture substrate was inoculated by pouring 15 ml of the conidial suspension of the n3 isolate (fusarium) at a concentration of 106 spores/ml into each pot containing sterile soil. the fungus was allowed to settle for 48 h in the growing medium. the previously disinfected chickpea seeds were sown into these pots at the rate of 5 seeds per pot. three replicates were prepared for each of the treatments (each pot was a replicate). two lots of chickpea cultivation pots (control and inoculated) were brought back to the greenhouse to promote seed germination, plant growth, and symptom development monitoring. after 4 weeks, plant emergence and plant survival were determined in pots containing inoculated seeds. in the pots with inoculated culture substrate according to the second inoculation technique, plant emergence and disease symptoms on chickpea plants were noted after 8 weeks, followed by po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 6 http://www.ncbi.nlm.nih.gov/blast assessment of disease severity by calculating the leaf damage index according to the scale established by douira and lahlou (1989). the scores related to the number of leaves constitute the foliar alteration index, calculated according to the formula below (douira & lahlou, 1989): fai = [σ(i×xi)] / 6×ntf fai: foliar alteration index. i: leaf appearance notes 0 – 5. xi: number of leaves with note i. ntf: total number of leaves. an average index was then calculated for each batch of plants. at the end of the trial, the pots were brought back to the laboratory to re-isolate the pathogen from the different parts of the plants (roots, crown, stems, and leaf petioles) obtained either from the inoculated seeds (technique 1) or inoculated growing substrate (technique 2). the different parts were separated and disinfected with 95% alcohol for 2 min. samples were then rinsed several times with sterile distilled water, dried quickly on sterile filter paper, transferred to psa medium, and incubated in the dark at 25 °c. the microscopic observation was performed after 1 week. notes appearance of leaves 0 healthy appearance 1 cotyledonary leaf: wilting or yellowing 2 cotyledonary leaf: fall 3 true leaf: wilting or yellowing 4 true leaf: necrosis 5 true leaf: fall po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 7 the re-isolation percentage (rp%) was calculated using the following formula: rp = (ns px /nt) × 100, where ns px is the number of segments containing the fungal species x and nt is the total number of segments used for re-isolation. after 8 weeks, the lengths of the aerial and root parts of the chickpea plant were measured with a double decimeter and the number of leaves and pods of each plant was counted. the fresh weights of the aerial and root parts were recorded using a precision balance and the dry weight of these parts was recorded after being dried in the oven at 70 °c for 48 h. 3. results the isolate n3 of fusarium equiseti was obtained from symptomatic samples among a complex of fusarium species including f. solani and f. oxysporum. the percentage of isolation was 3% in the case of f. equiseti. on the psa medium, the isolate n3 developed a colony with an abundant aerial mycelium which was fluffy and beige-white colored (figure 1). under a microscope, the mycelial filaments were septate. chlamydospores were present (7 to 13 μm in diameter, spherical, globular, most often intercalary, solitary,or in pairs, and frequently as short chains). macroconidia were numerous, slightly curved, usually with 5 to 6 septa and 31 to 45 μm long. the description was identical to that of f. equiseti (corda) saccardo, reported by leslie and summerell (2006) and rafique et al. (2019). the identity of the isolate n3 was evaluated and confirmed. after identification, the isolate n3 was registered in the national database under voucher id: rab111030 and submitted to genbank as accession no. mt111122. this sequence was 99.33% identical to fusarium equiseti. the results of the pathogenicity tests using two inoculation methods demonstrated the pathogenic capacity of the isolate n3 of f. equiseti towards a local variety of chickpea. few plants emerged po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 8 from seeds inoculated with the isolate n3. the majority of inoculated seeds turned rotten (figure 2) compared to un-inoculated seeds which grew normally under greenhouse conditions. furthermore, chickpea plants obtained from seeds inoculated with f. equiseti showed a disruption in the growth parameters. the length of the plants, average number of leaves produced as well as the average number of branches formed were much lower in plants from inoculated seeds than those presented by the control plants; these parameters were 1.29 and 29.05 cm, 1.11 and 24.21 leaves, and 0 and 3.50 twigs, respectively. fusarium equiseti isolate also adversely affected the growth of roots which exhibited a root length of 0.91 cm, and 0.78 and 0.13 g as fresh and dry weight of roots, respectively, in comparison with 27.16 cm, 7 g, and 5 g respectively, in the control plants. the symptoms observed in chickpea plants developed on a culture substrate inoculated with f. equiseti according to technique 2 were variable: necrosis of the roots and crown followed by wilting of the plants. after 8 weeks of cultivation, the aggressiveness of infection induced by the f. equiseti isolate as estimated by calculating the foliar alteration index was 0.395 in plants grown in the culture substrate inoculated with a conidial suspension of the fusarium isolate n3. regarding growth parameters, chickpea plants grown in the inoculated substrate displayed a length of 19.44 cm, an average of 18.66 leaves, and 2.08 twigs per plant compared to a length of 31.05 cm, 25.66 leaves, and 4.11 twigs per plant in control chickpea plants. plants which were grown in the culture substrate inoculated with f. equiseti also presented a lower root length, fresh and dry weight of root or aerial parts attaining 19.73 cm, 5.33 and 4.12 g, and 5.47 and 4.02 g, respectively, than plants grown in uninoculated soil (28.33 cm, 7.33 and 6.60 g, and 6.66 and 5.40 g, respectively). the re-isolation performed on the plants grown either from inoculated seeds or on soil infested with f. equiseti confirmed the presence of the fungus in different parts of the plants. using the po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 9 first technique, the highest percentage of the pathogen re-isolation was registered in the chickpea root with 70.22% followed by collar (70%), stem (60%), and leaf petiole (34.33 %), whereas percentages recorded in chickpea plants after soil inoculation were 84.77% (collar), 72.77% (root), 64.44% (stem), and 36.11% (petiole). fusarium equiseti was consistently reisolated from infected seeds and collar tissues, satisfying koch’s postulates. to our knowledge, this is the first report of fusarium equiseti causing seed rot and discoloration at the level of the crown and wilting of chickpea plants. fig. 1. characteristic of fusarium equiseti colony (a); chlamydospores (b); macroconidia and microconidia (c) magnification, 400×; mounting medium, lactophenol cotton blue. po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 10 fig. 2. symptoms of stunting in chickpea plants (a) derived from seeds inoculated with fusarium equiseti isolate (f.e) and control seedlings from non-inoculated chickpea seeds (t); b: symptoms of rot in inoculated chickpea seeds; c: brownish discoloration of crown in chickpea plants grown in f. equiseti-infested soil (f.e); d: chickpea plants grown in un-infested soil (t). 4. discussion the fusarium complex responsible for root rot and wilt diseases in chickpeas is diverse (zemoulipo st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 11 benfreha 2014). one of the representative candidates of this complex is the f. equiseti that was isolated for the first time in morocco from the roots of diseased chickpea plants and identified using morphological characters in addition to molecular characterization. in the current study, koch’s postulates were verified by inoculation of chickpea seeds and culture substrate. the inoculated seeds expressed weak germination and growing ability with a high degree of rotting and decomposition. chickpea plants from seeds sown in soil infested with a suspension of f. equiseti spores showed a range of deterioration symptoms such as root and crown necrosis, vascular browning, yellowing leaves, and stunting and wilting of the plants. the fungus was re-isolated from the roots, crown, stems, and leaf petioles of the inoculated plants through seed dip inoculation or soil infestation with the spore suspension of f. equiseti. the ability of the pathogen to invade the upper levels of plants can be inferred from these results. colonization of chickpea plants tissues by f. equiseti after inoculation affected their growth, leading to root and vegetative growth retardation including the production of leaves and twigs. all these types of symptoms were observed in chickpea plants inoculated with f. solani (el hazzat et al., 2019) as well as in lentil and cumin (cuminum cyminum) plants infected with f. equiseti (rafique et al., 2019; ramchandra et al., 2011). fusarium equiseti is among the fungal species capable of attacking several legumes species, in which it can induce damping-off and root rot disease (rubella et al., 2008). these authors have reported decayed seeds and reddish brown to black lesions on hypocotyl and roots of kidney bean (phaseolus vulgaris), pea (pisum sativum), and chickpea (cicer arietinum) following inoculation with f. equiseti isolates originating from fields of ginseng. similarly, the seeds that germinated in soil infected with f. equiseti resulted in plants that showed browning at the crown and stem base followed by wilting. some symptoms on leguminous hosts bore a resemblance to those stated herein. on wild pigeon pea, this fungal species provokes foliar po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 12 chlorosis, browning and black discoloration of the stem, plant drying, and ultimately plant death (mishra et al., 2021). fernandez and jefferson (2004) observed a discoloration in the subcrown stems and roots of cereal species. this fungal species is capable of infecting seeds, roots, tubers, and fruits of several species of cultivated plants, such as cucurbits (joffe & palti 1967), cotton (chimbekujwo, 2000), cowpea (rodrigues & menezes 2005), lentils (chaudhary & kaur 2002), sugar beet (stojsin et al., 2001), potatoes (rai, 1979; theron & holz 1989), citrus (sukmawati & miarsyah 2017), pine (ocamb & juzwik 1995), and even nursery plants (bloomberg, 1981). fusarium equiseti has also been isolated from cereals such as corn, wheat, and barley (ballois, 2012). this fungal species is responsible for rotting stems and premature wilting of corn plants (swamy et al., 2020) and root rot in winter wheat (booth 1971). sometimes, this pathogen is associated with blight of wheat ears (shaner, 2003; gale, 2003; tekauz et al., 2005; wing et al., 1993; xue et al., 2006) and rice panicles. infection can also occur during grain storage or afterward (hashem et al., 2010). in morocco, several fusarium species are associated with symptoms of root rot in cereals; the most common are f. equiseti, f. culmorum, f. oxysporum, and f. solani (lyamani 1988). fusarium equiseti, isolated for the first time in morocco, was isolated from the fungal complex associated with chickpea roots. pathogenicity tests conducted in the greenhouse showed that this fungal species is endowed with significant pathogenicity towards this host plant, and the symptoms developed were similar to those observed in chickpea plants inoculated with f. solani (el hazzat et al., 2019). extending the surveys to other regions of morocco is important to build a population of f. equiseti isolates and to determine the amplitude of the variation in the pathogenicity of this pathogen via other chickpea varieties grown in morocco. with time and in the absence of an po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 13 effective control program, fusarium equiseti will probably become an important pathogen and take its place among the other known diseases of chickpea. 5. conclusion to our knowledge, the present study showed for the first time that fusarium equiseti can play serious role in the disease process of chickpea in 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(1984). international testing of chickpeas for resistance to ascochyta blight. plant disease, vol. 68, p. 782. https://doi.org/10.1094/pd-69-782 swamy, s.d., mahadevakumar, s., hemareddy, h.b., amruthesh, k.n., mamatha, s., kunjeti s.g., swapnil, r., kumard, t.v., lakshmidevi, n., 2020. first report of fusarium equiseti– the incitant of post flowering stalkrot of maize (zea mays l.) in india. crop protection.129, 105-035. tekauz, a., stulzer, m., beyene, m., 2010. fusarium head blight of winter wheat in manitoba in 2009. canadian plant disease survey = inventaire des maladies des plantes au canada, p. 47– 48. theron, d.j., holz, g., 1989. fusarium species associated with dry and stem-end rot of potatoes in south africa. phytophylactica 21, 175–181. white, t.j., bruns, t., lee, s., taylor, j., 1990. amplification and direct sequencing of fungal ribosomal rna for phylogenetics. in: ma innes, dh gelfand, jj sninsky, tj white eds, po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 20 pcr protocols. a guide to methods and applications, academic press, inc., san diego, california, 315-322. xue, a.g., ho, k.m., butler, g., vigier, b.j, babcock, c., 2006. pathogenicity of fusarium species causing head blight in barley. phytoprotection 87, 55–61. zemouli-benfreha, f., henni, d-e., merzoug, a., 2014. fusarium wilt of chickpea (cicer arietinum l.) in north-west algeria. african journal of agricultural research 9, 168-175. po st pr in t vol. 57, article 576 doi: 10.5586/am.576 acta mycologica: postprint version 21 1. introduction 2. materials and methods 2.1 fungal material 2.2 morphological characterization 2.3 molecular analysis and identification 3. results 4. discussion 5. conclusion professor dr hab. maria lisiewska 1 of 3published by polish botanical society acta mycologica scientists professor dr hab. maria lisiewska małgorzata stasińska* department of botany and nature conservation, center for molecular biology and biotechnology, environmental testing laboratory, university of szczecin, felczaka 3c, 71-412 szczecin, poland * email: stasinsk@univ.szczecin.pl abstract the article presents the biography and scientific achievements of professor maria lisiewska. she earned master’s degree and ph.d. in natural sciences from adam mickiewicz university in poznań. after earning her doctoral degree, she stayed at adam mickiewicz university in poznań and conducted her thrilling research on mycology and taught until now. prof. maria lisiewska is an author of many books, articles, and other scholarly reports. keywords mycology; mycocoenology; macromycetes; poznań this issue of acta mycologica is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. professor maria lisiewska’s career has been inextricably linked with poznań, the town of her birth. she studied biology at the faculty of botany and earth sciences of adam mickiewicz university between 1952 and 1957. her thesis, “systemy korzeniowe roślin grądowych a podłoże [root systems of galio-carpinetum plants in dependence on the substratum],” was supervised by professor zygmunt czubiński. in 1964, she was awarded a doctorate in natural sciences from adam mickiewicz university upon submission of her dissertation, “udział grzybów wyższych w grądach wielkopolski [higher fungi of the querco-carpinetum of the wielkopolska province]” [1], written under the direction of professor zygmunt czubiński and research tuition of professor andrzej nespiak. the result of her postdoctoral research work (habilitation) was entitled “grzyby wyższe lasów bukowych we wschodniej części zasięgu buka w europie [macromycetes of beech forests within the eastern part of the fagus area in europe]” [2]. maria lisiewska received a professorial nomination award for extraordinary professorship in 1985 and the position of full academic professor in 1993. her first teaching and research job was also at adam mickiewicz university, where, while still a student, she was employed as a student assistant in the department of plant systematics and geography (at present the department of plant ecology and environment conservation), chaired by professor czubiński, on december 1, 1955. thanks to the kindness of professor czubiński, maria lisiewska developed her mycological passion and interest in fungi by attending mycological and phytopathological seminars under the guidance of professor karol mańka and professor karol zaleski at the higher school of agriculture. she was subsequently employed as a junior lecturer (from 1957), senior lecturer (from 1964), associate professor (docent, doi: 10.5586/am.1061 publication history received: 2015-06-27 accepted: 2015-07-31 published: 2015-08-05 handling editor maria rudawska, institute of dendrology of the polish academy of sciences, poland funding this work did not involve any funding. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation stasińska m. professor dr hab. maria lisiewska. acta mycol. 2015;50(1):1061. http://dx.doi. org/10.5586/am.1061 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:stasinsk%40univ.szczecin.pl?subject=professor%20dr%20hab.%20maria%20lisiewska http://dx.doi.org/10.5586/am.1061 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1061 http://dx.doi.org/10.5586/am.1061 2 of 3© the author(s) 2015 published by polish botanical society acta mycol 50(1):1061 stasińska / professor dr hab. maria lisiewska from 1974), extraordinary professor (from 1985), and full professor (ordinary, from 1993) until her retirement in 2004. in july 2008, maria lisiewska was awarded the status of professor-emeritus (profesor-senior) at her alma mater. maria lisiewska’s research output comprises 97 original studies, both research papers and monographs, published in academic journals in poland and abroad. she is the author of a monograph study of the genus mycena, based on specimens from the entire area of poland and illustrated with color drawings from nature of mycena made by professor maria lisiewska. the study was published as the seventeenth volume of flora polska: grzyby (mycota) [the flora of poland: fungi (mycota)], a publication series by the institute of botany, polish academy of sciences. together with marian szmid, from the provincial sanitary and epidemiological station (sanepid) in poznań, lisiewska wrote the popular book przewodnik grzyboznawczy [guide to mushrooms], which has now had its fifth edition. she is also the coauthor of the mycology textbook mikologia: przewodnik do ćwiczeń terenowych i laboratoryjnych [guide for field and laboratory exercises] [3]. professor maria lisiewska’s interests lie mostly in the mycocoenology and ecology of macromycetes. she has conducted research in a variety of forest communities: oakhornbeam forests, beech forests, acidophilous oak forests, and coniferous forests. she is interested in the species diversity of fungi in relation to different plant communities. the region of her native wielkopolska as well the western bieszczady mountains, świętokrzystkie mountains, masurian lakeland, and pomerania are at the center of her investigations. professor lisiewska has taken part in numerous research programs, such as the interdisciplinary crypto project (“cryptogamous plants in the forest communities of białowieża national park”) coordinated by professor janusz b faliński, the international project on “mycological monitoring in european oak forests” coordinated by professor maria ławrynowicz, and complex mycocoenological investigations in germany, denmark, and many countries of the former yugoslavia. she has presented the results of her research work at european and international mycological congresses, from the fourth congress of european mycologists in warsaw (poland) in 1966 to the fourteenth congress of european mycologists in katsiveli (crimea, ukraine) in 2003, as well as many conferences in poland. since 1982, professor maria lisiewska has been the science editor of the research journal badania fizjograficzne, seria b: botanika [physiographic studies, series b: botany], published by the poznańskie towarzystwo przyjaciół nauk [poznań society of friends of sciences], until 2010 badania fizjograficzne nad polską zachodnią, seria b [physiographic studies of western poland, series b]. she has also, since 1995, been a member of the editorial board of the international journal acta mycologica. professor lisiewska’s professional affiliations include membership to the mycological section of the polish society of hygiene (a member since 1965 and a member of the executive board), the polish botanical society, and the polish mycological society, of which she is a founding member. professor maria lisiewska has acted as director of extramural studies in biology (1978–1985) and vice dean of the faculty of biology (1985–1987). she was a visiting professor at the university of bologna in june 1992. professor lisiewska worked as an academic teacher at the faculty of biology at adam mickiewicz university until 2004, although she continued delivering lectures until 2013. she has supervised as many as nineteen undergraduate dissertations, sixty master’s theses, and two doctoral theses (stefan friedrich and grzegorz fiedorowicz). she has acted as a reviewer–examiner of seven applications for the academic title of professor and eight postdoctoral theses (habilitation). for over fifty years, maria lisiewska has conducted highly popular and equally respected lectures and field seminars at annual courses devoted to fungi organized by the state sanitary and epidemiological inspectorate in poznań. as an outstanding academic teacher and educator, professor maria lisiewska has received numerous awards and distinctions, such as individual prizes from the ministers of education, higher education and technology (1975) and national education (1988), a joint prize from the minister of nature protection, natural resources and forestry (1999), and a gold medal celebrating the hundredth anniversary of the polish society of hygiene (1998). 3 of 3© the author(s) 2015 published by polish botanical society acta mycol 50(1):1061 stasińska / professor dr hab. maria lisiewska references 1. lisiewska m. higher fungi of the querco-carpinetum of wielkopolska province. acta mycol. 1965;1(1):169–271. http://dx.doi.org/10.5586/am.1965.012 2. lisiewska m. macromycetes of beech forests within the eastern part of the fagus area in europe. acta mycol. 1974;10(1)3–72. http://dx.doi.org/10.5586/am.1974.001 3. bujakiewicz a, lisiewska m, nita j. mikologia: przewodnik do ćwiczeń terenowych i laboratoryjnych. poznań: bogucki wyd. nauk.; 2007. http://dx.doi.org/10.5586/am.1965.012 http://dx.doi.org/10.5586/am.1974.001 abstract references 2015-08-02t15:16:38+0100 piotr otręba the revision of specimens of the cladonia pyxidata-chlorophaea group (lichenized ascomycota) from northeastern poland deposited in the herbarium collections of university in bialystok 1 of 11published by polish botanical society acta mycologica original research paper the revision of specimens of the cladonia pyxidata-chlorophaea group (lichenized ascomycota) from northeastern poland deposited in the herbarium collections of university in bialystok anna matwiejuk* department of plant ecology, institute of biology, university of bialystok, konstantego ciołkowskiego 1j, 15-245 bialystok, poland * email: matwiej@uwb.edu.pl abstract in northeastern poland, the chemical variation of the cladonia chlorophaea-pyxidata group was much neglected, as tlc has not been used in delimitation of species differing in the chemistry. as a great part of herbal material of university in bialystok from ne poland was misidentified, i found my studies to be necessary. based on the collection of 123 specimens deposited in herbarium of university in bialystok, nine species of the c. pyxidata-chlorophaea group are reported from ne poland. the morphology, secondary chemistry, and ecology of examined lichens are presented and the list of localities is provided. the results revealed that c. fimbriata is the most common species in the northeastern poland, comprising around 33% of the studied specimens. cladonia conista, c. cryptochlorophaea, and c. merochlorophaea are known only from very few locations. this study shed light on the role of the lichens substances to diagnosis of the species of c. pyxidatachlorophaea group. keywords cladoniaceae; chemotaxonomy; distribution; ecology introduction lichens of the cladonia pyxidata-chlorophaea group are characterized by primary thallus squamulose and scyphose podetia. the podetia are covered with farinose to granular soredia, corticated granules, and/or more or less areolate cortex. apothecia are brown and rare [1,2]. the species of this group have a diverse secondary chemistry. fourteen chemotypes and 35 lichen secondary metabolites have been identified within the group from the world [3]. however, the species status of chemically different entities has been frequently questioned. some authors recognize chemical variability only at the chemotypes of species, varieties and subspecies level [4–6], others support chemically defined taxa at the species level [2,7–9]. many authors accept the later viewpoint about chemically species [2,9–12]. the c. pyxidata-chlorophaea group requires molecular investigation with a larger number of samples and more variable gene regions [10,11]. the recent molecular studies have indicated that at least all the chemically different taxa of the c. chlorophaea group do not form a separate subclade and their morphological similarity is rather a result of convergent evolution [10]. doi: 10.5586/am.1087 publication history received: 2016-04-02 accepted: 2016-12-21 published: 2017-01-16 handling editor maria rudawska, institute of dendrology, polish academy of sciences, poland funding research funded by the polish ministry of science and higher education within the statutory research. competing interests no competing interests have been declared. copyright notice © the author(s) 2017. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation matwiejuk a. the revision of specimens of the cladonia pyxidata-chlorophaea group (lichenized ascomycota) from northeastern poland deposited in the herbarium collections of university in bialystok. acta mycol. 2016;51(2):1087. http:// dx.doi.org/10.5586/am.1087 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:matwiej%40uwb.edu.pl?subject=the%20revision%20of%20specimens%20of%20the%20cladonia%20pyxidata-chlorophaea%20group%20%28lichenized%20ascomycota%29%20from%20northeastern%20poland%20deposited%20in%20the%20herbarium%20collections%20of%20university%20in%20bialystok http://dx.doi.org/10.5586/am.1087 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1087 http://dx.doi.org/10.5586/am.1087 2 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland the aim of the present study is the revision of specimens of the c. pyxidata-chlorophaea group (lichenized ascomycota) from the area of ne poland deposited in herbarium of university in bialystok. for each specimen examined, the occurrence of the lichens of the c. pyxidata-chlorophaea group from tested area along with their morphology, chemistry, ecology, and distribution of each species is described. the chemical variation of the c. pyxidata-chlorophaea group from ne poland was not sufficiently recognized because thin layer chromatography (tlc) has not been used during species identification. the area of study includes protected areas (e.g., wigry national park, biebrza national park, narew national park, podlaski przełom bugu landscape park, puszcza knyszyńska landscape park) and areas not protected in the vicinity of the villages or small towns (e.g., suchowola, kaniuki, ciechanowiec, boćki) and surrounding the siemianówka reservoir in the upper narew valley and forests (e.g., romincka forest). material and methods the revised lichen material was collected in ne poland by abramowicz a., bystrek j., czubała a., gosk a., gutowska m., jabłońska k., karpowicz a., kolanko k., kulikowska k., matwiejuk a., pietryszek m., śliwowoska j., świderska m., zabuska s., in the years 1991–2015. the morphology of each specimen was examined using a stereomicroscope (leica ez4) at magnification 0.8(–3.5) × 10. lichen substances were investigated by tlc in solvent a and c following the methods of orange et al. [13]. in total, 123 specimens were examined. results nine species of the c. pyxidata-chlorophaea group were found in the examined material. cladonia fimbriata appeared to be the most common species of the investigated lichen group in ne poland. three taxa, c. conista, c. cryptochlorophaea, c. merochlorophaea are known only from very few locations. cladonia chlorophaea (flörke ex sommerf.) spreng., syst. veg. 4: 273(1827) a characteristic species with regular podetia covered soredia. podetia are up to ca. 3.0 cm tall. scyphi are up to 7 mm wide, with irregular or dentate margins, usually gradually expanded. scyphi and surface of podetia covered by granular soredia and corticated granules. apothecia are brown and stalked, on cup margin. pycnidia occur on scyphal margins. for a detailed description see [5,14–16]. the species is characterized by the production of fumarprotocetraric acid complex only. the specimens of c. chlorophaea are often morphologically similar to c. grayi and c. merochlorophaea. the species is common and inhabits a many substrata, namely soil (12 specimens), bark (six specimens) and wood (four specimens). corticolous specimens were collected from betula spp. (two specimens), pinus sylvestris (one specimen), salix spp. (one specimen), fraxinus excelsior (one specimen), and robinia pseudoacacia (one specimen). robinia pseudoacacia is a new phorophyte of c. chlorophaea in ne poland [2]. a similar habitat requirement was also discovered from poland by kowalewska et al. [2] and belarus by tsurykau and golubkov [9]. location of investigation sites of c. chlorophaea in ne poland from the herbarium collections of university of bialystok has been presented in fig. 1.   białystok podlaskie mazowieckie lubelskie suwalski pk warmińsko mazurskie pk puszczy knyszyńskiej łomżyński pk doliny narwi pk podlaski przełom bugu fig. 1. location of investigation sites of cladonia chlorophaea narew bie brz a bug fig. 1 location of investigation sites of cladonia chlorophaea. 3 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland cladonia chlorophaea is widely distributed in poland [2]. many localities of this species were reported in ne poland by kowalewska et al. [2]. it is known from all continents including the antarctic region [2,8,17,18]. specimens examined. ne poland, podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-05-05, det. a. matwiejuk, 2014-11-16; surrounding of the siemianówka lagoon, village bondary, soil, leg. b. marszalik, 1993-06-17, det. a. matwiejuk, 2014-11-16; romincka forest, spruce-pine forest, soil, leg. m. gutowska, 1991-07-26, det. a. matwiejuk, 2014-11-16; romincka forest, spruce-pine forest, soil, 1991-07-28, leg. m. gutowska, det. a. matwiejuk, 2014-11-16; romincka forest, spruce-pine forest, soil, 1992-09-19, leg. m. gutowska, det. a. matwiejuk, 2014-11-16; romincka forest, spruce-pine forest, soil, leg. m. gutowska, 1992-09-12, det. a. matwiejuk, 2014-11-16; klejniki, logging, soil, leg. m. pietryszek, 1999-05-26, det. a. matwiejuk, 2014-11-16; klejniki, logging, the botttom, soil, leg. m. pietryszek, 1999-05-26, det. a. matwiejuk, 2014-11-16; klej niki, logging, western slope, soil, leg. m. pietryszek, 1999-05-4, det. a. matwiejuk, 2014-11-16; narew national park, bark of robinia pseudoacacia, leg. a. mrozowska, 2001-10-24, det. a. matwiejuk, 2014-11-16; narew national park, bark of fraxinus excelsior, leg. a. mrozowska, 2001-01-16, det. a. matwiejuk, 201411-16; surroundings of suraż village, doktorce, soil, leg. k. jabłońska, 2001-01-16, det. a. matwiejuk, 2014-11-16; surroundings of suraż village, doktorce, wood, leg. k. jabłońska, 2002-05-14, det. a. matwiejuk, 2014-11-16; surroundings of suraż village, doktorce, approx. 0.5 km to the nw, pine forest, soil, leg. k. jabłońska, 2002-05-14, det. a. matwiejuk, 2014-11-16; surroundings of suraż village, doktorce, wood, building a farm, leg. k. jabłońska, 2001-09-20, det. a. matwiejuk, 2014-11-16; surroundings of suraż village, doktorce, bark of betula pendula, leg. k. jabłońska, 2002-08-13, det. a. matwiejuk, 2014-11-16; surroundings of ciechanowiec, forest by the nurzec river, village zadobrze, soil, leg. s. zabuska, 2004-11-7, det. a. matwiejuk, 2014-11-16; surroundings of laskowiec village, wood, leg. s. zabuska, 2014-11-16. det. a. matwiejuk, 2014-11-16; surroundings of laskowiec village, soil, leg. a. gosk, 1999-11-01, det. a. matwiejuk, 2014-11-16; surroundings of laskowiec village, soil, leg. a. gosk, 199911-01, det. a. matwiejuk, 2014-11-16; surroundings of lasko wiec village, soil, leg. a. gosk, 1999-11-01, det. a. matwiejuk, 2014-11-16; laskowiec stary (near zambrów), bark of salix sp. wood, leg. a. gosk, 2000-11-11, det. a. matwiejuk, 2014-11-16; zamczysk, bark of betula pendula, leg. a. gosk, 2001-06-6, det. a. matwiejuk, 2014-1116; ciechanowiec, soil, leg. s. zabuska, 2004-05-26, det. a. matwiejuk, 2014-11-16; boćki, soil, leg. a. matwiejuk, 2001-03-30, det. a. matwiejuk, 2014-11-16; forest near rykaczew, soil, leg. a. gosk, 2000-07-11, det. a. matwiejuk, 2014-11-16; surrounding of laskowiec village, forest, bark of pinus sylvestris, leg. a. gosk, 1999-11-01, det. a. matwiejuk, 2014-11-16; puszcza knyszyńska landscape park, góry leńce, young forest pine, soil, leg. k. kulikowska, 1992-08-25, det. a. matwiejuk, 2014-11-16. number of specimens examined: 29. cladonia conista robbins ex a. evans, trans. connecticut acad. arts 30: 472 (1930) cladonia conista is distinguished by the podetia tall, up to ca. 2.5 cm high, with regular, ± goblet-shaped cups. the upper part of the stalk and the cups with soredia. the lower part of the stalk is corticated. soredia are farinose rather than granular. the detailed description of the species is presented elsewhere [2,7]. secondary metabolites of c. conista include bourgeanic and fumarprotocetraric acids. cladonia conista is morphologically very similar to c. humilis. in contrast to this species, c. conista produces bourgeanic acid (k−). cladonia humilis produces fumarprotocetraric acid and atranorin (k+ yellow). pino-bodas et al. [12] have stated that the species are morphologically indistinguishable, but the taxa differ in their geographical distribution and c. conista and c. humilis are recognized as distinct species. the species was found on soil in pine woodland (one specimen). the observed habitat requirements agreed with those reported for the species by kowalewska et al. [2]. all the polish specimens were found on soil. distribution of stands of c. conista 4 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland in ne poland from the herbarium collections of university of bialystok has been presented in fig. 2. cladonia conista is rare species in poland and ne poland [2]. it is predominant on the atlantic coast of america and in northern regions of europe [2,9,12]. specimens examined. ne poland, surrounding of the siemianówka reservoir in the upper narew valley, pine forest, soil, det. a. matwiejuk, 2014-05-12, leg. a. matwiejuk, 2015-05-23. number of specimens examined: 1. cladonia cryptochlorophaea asahina, j. jap. bot. 16: 711 (1940) this species is characterized by the podetia gobletto trumpetshaped cups. soredia usually present. the surface of the podetia is mostly roughly corticated. for the description of the species see [2,7,15]. substances detected by tlc include cryptochlorophaeic, paludosic, and fumarprotocetraric acids. cladonia cryptochlorophaea is morphologically very similar to few species (c. merochlorophaea, c. novochlorophaea, c. grayi) [1,2,7], but the latter differs chemically by producing cryptochlorophaeic and paludosic acids. morphologically, c. cryptochlorophaea resembles c. grayi, but the podetia of cladonia grayi are usually abundantly granular-sorediate [2]. substrate of the specimen includes soil. the species prefers pine forests. almost three-quarters of the records of c. cryptochlorophaea in poland [2] are represented by epigeic specimens. the similar ecology notes are given by holien and tønsberg [7], ahti [15]. distribution of investigation site of this species in ne poland from the herbarium collections of university of bialystok has been presented in fig. 3. in poland, it is rare species [2], but its known localities are scattered, mostly in northeastern and southern part of poland [2,19]. the species is known from all continents, except antarctica. many specimens have been recorded mostly in boreal zone [9]. specimens examined. ne poland, surrounding of the siemianówka reservoir in the upper narew valley, pine forest, soil, det. a. matwiejuk, 2014-05-12, leg. a. matwiejuk, 2015-05-23. number of specimens examined: 1. cladonia fimbriata (l.) fr., lichenogr. eur. ref.: 222 (1831) this species is distinguished by the podetia up to 3.0 cm tall, simple, with scyphi. scyphi are up to 6 mm wide, regular. podetia with goblet-shaped. the surface of podetia and scyphi coated with soredia. soredia are farinose. apothecia are rare, brown and simple. for the description of the species see [2,5,14–16]. the species produces fumarprotocetraric acid complex only. cladonia fimbriata may resemble c. chlorophaea. it has more regular and extended scyphi, granular soredia and smaller stalks. cladonia fimbriata is most similar to c. conista, but c. conista differs by its usually distinctly corticate podetial stalk [2,20]. the species is ubiquitous and inhabits a wide range of substrata. in ne poland, c. fimbriata prefers well-lit open pine, oak and birch forests and urban areas. the species was found on soil (30 specimens), wood (eight specimens), betula spp. (one specimen), pinus sylvestris (one specimen), rock (one specimens). location of investigation sites of c. fimbriata in ne poland has been presented in fig. 4.   białystok podlaskie mazowieckie lubelskie suwalski pk warmińsko mazurskie pk puszczy knyszyńskiej łomżyński pk doliny narwi pk podlaski przełom bugu fig. 2. location of investigation site of cladonia conista narew bie brz a bug fig. 2 location of investigation site of cladonia conista.   białystok podlaskie mazowieckie lubelskie suwalski pk warmińsko mazurskie pk puszczy knyszyńskiej łomżyński pk doliny narwi pk podlaski przełom bugu fig. 3. location of investigation site of cladonia cryptochlorophaea narew bie brz a bug fig. 3 location of investigation site of cladonia cryptochlorophaea. 5 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland cladonia fimbriata is common species in poland [2]. many localities of this species were reported in ne poland by kowalewska et al. [2]. outside poland, the species has been recorded from many regions. it is known from all continents except for tropic regions [8,18]. specimens examined. ne poland, hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, the embankment stones, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, the embankment stones, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-1012, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, the embankment stones, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-1116; hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-1012, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of birch, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of pine and spruce, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of pine and spruce, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of pine and spruce, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; hołodolina, 0.5 km s, edge of the forest of pine and spruce, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 0.5 km ne, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 201411-16; suchowola, 1 km ne, pine forest, soil, leg. a. abramowicz, det. 2014-10-12, a. matwiejuk, 2014-11-16; suchowola, 1 km ne, pine forest, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 1 km ne, pine forest, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 1 km ne, pine forest, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-1116; suchowola, 1 km ne, pine forest, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 1 km ne, pine forest, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 1 km ne, pine forest, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 1 km ne, pine forest, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-1116; boćki, soil, leg. a. matwiejuk, 2000-04-17, leg. a. matwiejuk, 2014-11-16; puszcza knyszyńska, krasne reserve, soil, leg. j. śliwska, 1992-07-29, det. a. matwiejuk, 201411-16; surrounding of suraż, soil, leg. k. jabłońska, 2002-08-16, det. a. matwiejuk, 2014-11-16; surrounding of ciechanowiec, wood, leg. s. zabuska, 2003-07-18, det. a. matwiejuk, 2014-11-16; surrounding of ciechanowiec, wall, leg. s. zabuska, 2004-1020, det. a. matwiejuk, 2014-11-16; surrounding of ciechanowiec, soil, leg. s. zabuska, 2003-09-2, det. a. matwiejuk, 2014-11-16; surrounding of ciechanowiec, soil, leg. s. zabuska, 2003-07-01, det. a. matwiejuk, 2014-11-16; surrounding of ciechano wiec, bark of betula pendula, leg. s. zabuska, 2004-07-11, det. a. matwiejuk, 2014-11-16; surrounding of ciechanowiec, soil, leg. s. zabuska, 2003-09-02, det. a. matwiejuk,   białystok podlaskie mazowieckie lubelskie suwalski pk warmińsko mazurskie pk puszczy knyszyńskiej łomżyński pk doliny narwi pk podlaski przełom bugu fig.4. location of investigation sites of cladonia fimbriata narew bie brz a bug fig. 4 location of investigation sites of cladonia fimbriata. 6 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland 2014-11-16; surrounding of laskowiec village, wood, leg. a. gosk, 1991-11-01, det. a. matwiejuk, 2014-11-16; surrounding of laskowiec village, soil, leg. a. gosk, 199111-01, det. a. matwiejuk, 2014-11-16; puszcza knyszyńska landscape park, biele, soil, leg. k. kolanko, 1993-07-15, det. a. matwiejuk, 2014-11-16; stryki, soil, leg. m. świderska, 2004-06-25, det. a. matwiejuk, 2014-11-16. number of specimens examined: 41. cladonia grayi g. merr. ex sandst., clad. exs. no. 1847 (1929) the squamulose of primary thallus are shrub-like. the podetia are up to 4.0 cm tall. in the upper part, the podetia are covered by granular soredia, with goblet or trumpetshaped scyphi up to ca. 1 cm wide, often with proliferations. apothecia are brown to dark brown and stalked. for a detailed description see [2,5,7]. one chemotypes is recognized in ne poland: grayanic acid always and additionally substances of the fumarprotocetraric acid complex (chemotype ii). in poland, kowalewska et al. [2] noted 81% specimens from chemotype ii. in nordic countries, chemotype i is somewhat more frequent [8]. in belarus, both chemotype i (grayanic acid always by accompanied 4-o-demethylgrayanic acid) and ii are similarly frequent [9]. the species is similarly to c. merochlorophaea and c. novochlorophaea. differs by producing grayanic acid [16]. cladonia grayi is mostly found in pine forests. the species inhabits soil (20 specimens) and wood (six specimens). distribution of stands of c. grayi in ne poland has been presented in fig. 5. in poland, c. grayi is common species [2]. many localities of this species were reported in ne poland by kowalewska et al. [2]. world distribution data of c. grayi are most widely, it has been reported from many continents: europe, asia, north america, central america, south america, australia, and new zealand [2,8]. specimens examined. ne poland, podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-05-05, det. a. matwiejuk, 2014-11-16; podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-0505, det. a. matwiejuk, 2014-11-16; podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-05-05, det. a. matwiejuk, 2014-11-16; podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-05-05, det. a. matwiejuk, 2014-11-16; podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-05-05, det. a. matwiejuk, 2014-11-16; podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-05-05, det. a. matwiejuk, 2014-11-16; podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-04-23, det. a. matwiejuk, 2014-11-16; podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-04-23, det. a. matwiejuk, 2014-11-16; podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2002-05-05, det. a. matwiejuk, 2014-11-16; puszcza knyszyńska landscape park, góry leńce, young forest pine, soil, leg. k. kulikowska, 1992-0825, det. a. matwiejuk, 2014-11-16; puszcza knyszyńska landscape park, sandy slope, soil, leg. k. kulikowska, 1991-08-27,   białystok podlaskie mazowieckie lubelskie suwalski pk warmińsko mazurskie pk puszczy knyszyńskiej łomżyński pk doliny narwi pk podlaski przełom bugu fig. 5. location of investigation sites of cladonia grayi narew bie brz a bug fig. 5 location of investigation sites of cladonia grayi. 7 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland det. a. matwiejuk, 2014-11-16; puszcza knyszyńska landscape park, sandy slope, soil, leg. k. kulikowska, 1991-08-27, det. a. matwiejuk, 2014-11-16; biebrza national park. gugny, swampy forest, leg. k. kolanko, 1993-06-22, det. a. matwiejuk, 201411-16; biebrza national park. gugny, swampy forest, leg. k. kolanko, 1993-06-22, det. a. matwiejuk, 2014-11-16; biebrza national park. gugny, swampy forest, leg. k. kolanko, 1993-06-22, det. a. matwiejuk, 2014-11-16; biebrza national park. gugny, swampy forest, leg. k. kolanko, 1990-05-10, det. a. matwiejuk, 2014-11-16; wigry national park, studziany las reserve, soil, leg. j. bystrek, a. matwiejuk, 1993-08-06, det. a. matwiejuk, 2014-11-16; surroundings of ciechanowiec, wood, leg. s. zabuska, 2003-07-10, det. a. matwiejuk, 2014-11-16; boćki, soil, leg. a. matwiejuk, 2000-0417, det. a. matwiejuk, 2014-11-16; suchowola, 0.5 km ne, pine forest, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 0.5 km ne, pine forest, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 0.5 km ne, pine forest, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 0.5 km ne, pine forest, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 0.5 km ne, pine forest, wood, leg. a. abramo wicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 0.5 km ne, pine forest, wood, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 1 km ne, pine forest, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16. number of specimens examined: 26. cladonia merochlorophaea asahina, j. jap. bot. 16: 713 (1940) podetia are up to ca. 3 cm tall, simple, with gradually flaring scyphi. scyphi are up to 1.5 cm wide, usually areolate-corticate, verrucolose, covered by coarse granules. the granules may change into phyllidia, microsquamules or macrosquamules. surface of podetia are areolate-corticate. apothecia are brown and stalked. revised specimens from the detailed description by holien and tønsberg [7], ahti [14], and kowalewska et al. [2]. substances detected by tlc are merochlorophaeic and 4-o-methylcryptochlorophaeic acids and fumarprotocetraric acid complex (chemotype ii). in poland, 90% of specimens produce fumarprotocetraric acid [2]. in belarus, 67% specimens contain this acid [9]. holien and tønsberg [7] reported a similar proportion from norway. morphologically, c. merochlorophaea is similar to the other species, e.g., c. cryptochlorophaea, c. novochlorophaea, but the latter differs chemically by producing cryptochlorophaeic acid. cladonia merochlorophaea is found in pine forests, on soil (two specimens). the similar ecology notes are given by holien and tønsberg [7] and kowalewska et al. [2]. location of investigation site of c. merochlorophaea in ne poland from the herbarium collections of university of bia lystok has been presented in fig. 6. occurrence of c. merochlorophaea has been reported from many localities of poland including several sites of north-eastern part of the country [2]. the species is known from all continents, except antarctica [2,18]. specimens examined. ne poland, suchowola, 1 km ne, pine forest, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16; suchowola, 1 km ne, pine forest, soil, leg. a. abramowicz, 2014-10-12, det. a. matwiejuk, 2014-11-16. number of specimens examined: 2.   białystok podlaskie mazowieckie lubelskie suwalski pk warmińsko mazurskie pk puszczy knyszyńskiej łomżyński pk doliny narwi pk podlaski przełom bugu fig. 6. location of investigation site of cladonia merochlorophaea narew bie brz a bug fig. 6 location of investigation site of cladonia merochlorophaea. cladonia merochlorophaea asahina, j. jap. bot. 16: 713 (1940) 8 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland cladonia monomorpha aptroot, sipman & van herk, lichenologist 33: 273 (2001) squamules of primary thallus are greenish-grey, large and thick. podetia are up to 1.5 cm tall, with regular cups. scyphi are gradually flaring, inside covered with bullate corticate plates inside. apothecia are brown, simple or occur in glomerulose accumulations. for detailed descriptions see tsurykau and golubkov [9], aptroot et al. [21], and kowalewska and kukwa [22]. the species is characterized by the production only fumarprotocetraric acid complex. cladonia monomorpha is similar to c. pyxidata and c. pocillum. all those taxa are esorediate and contain fumarprotocetraric acid as the mayor secondary metabolite, while they differ morphologically. for a detailed description see kowalewska et al. [2] and aproot et al. [21]. in ne poland, the species was found on soil (one specimen). in poland, the frequency on different substrata is as follows: soil, rocks, and tree bark [2]. this agrees with the habitat spectrum published by aproot et al. [21]. location of investigation site of c. monomorpha from the herbarium collections of university of bialystok has been presented in fig. 7. most of the records come from the northern and southern part of the country [2]. several localities of c. monomorpha were reported in ne poland by kowalewska et al. [2]. the species is known from europe, asia, and north america [2,21]. specimens examined. ne poland, kaniuki, 1 km se, pine forest, soil, leg. a. matwiejuk, 2015-10-10, det. a. matwiejuk, 2015-11-23. number of specimens examined: 1. cladonia novochlorophaea (sipman) brodo & ahti, canad. j. bot. 74: 1167 (1996) squamules are brown-grey. podetia are brownish or blackish, up to 1.4 cm tall. scyphi are up to 0.8 cm wide, simple to proliferating from margins. surface of podetia are corticate, verruculose, squamulose, inner part of the scyphi eroded, covered with cortical plates. apothecia are stalked, brown. for detailed descriptions see brodo and ahti [5], tsurykau and golubkov [9], and ahti [14]. the species is characterized by the production homosekikaic, sekikaic, and fumarprotocetraric acids. cladonia novochlorophaea is morphologically similar to c. merochlorophaea, but it is never clearly sorediate. it has a similar chemistry to c. homosekikaica. in ne poland, c. novochlorophaea has been found on soil (six specimens), wood (three specimens) and bark of picea abies (one specimen) in open habitats and forests. distribution of stands of this species in ne poland has been presented in fig. 8. it is known from northern and central part of the poland. it is rare species. three localities of c. novochlorophaea were reported in ne poland by kowalewska et al. [2]. the species has been reported from europe, north and south america, new zealand, and the antarctic region [8,23]. specimens examined. ne poland, podlaski przełom bugu landscape park, forest district sarnaki, forestry mierzwice, pine forest, soil, leg. m. murawska, b. bystrek, 2001-05-05, det. a. matwiejuk, 2014-12-10; puszcza knyszyńska landscape park, kopisk, bark of picea abies, leg. k. kulikowska, 1992-05-12, det. a. matwiejuk, 2014-12-10; puszcza knyszyńska landscape park,   białystok podlaskie mazowieckie lubelskie suwalski pk warmińsko mazurskie pk puszczy knyszyńskiej łomżyński pk doliny narwi pk podlaski przełom bugu fig. 7. location of investigation site of cladonia monomorpha narew bie brz a bug fig. 7 location of investigation site of cladonia monomorpha.   białystok podlaskie mazowieckie lubelskie suwalski pk warmińsko mazurskie pk puszczy knyszyńskiej łomżyński pk doliny narwi pk podlaski przełom bugu fig. 8. location of investigation sites of cladonia novochlorophaea narew bie brz a bug fig. 8 location of investigation sites of cladonia novochlorophaea. 9 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland surroundings of waliły village, young forest birch, soil, leg. a. karpowicz, 1991-0827, det. a. matwiejuk, 2014-12-10; biebrza national park, gugny, pine forest, soil, leg. k. kondej, 1990-10-27, det. a. matwiejuk, 2014-12-10; puszcza knyszyńska landscape park, zalesie, soil, leg. k. kulikowska, 1991-10-26, det. a. matwiejuk, 2014-12-10; rybniki, wood, leg. m. świderska, 2004-06-25, det. a. matwiejuk, 201412-10; rybniki, wood, leg. b. marszalik, 1992-05-19, det. a. matwiejuk, 2014-12-10; surroundings of ciechanowiec, forest by the nurzec river, village zadobrze, soil, leg. s. zabuska, 2003-10-10, det. a. matwiejuk, 2014-12-10; laskowiec stary (near zambrów), wooden house, leg. a. gosk, 1999-11-11, det. a. matwiejuk, 2014-1210; biebrzańska kotlina, grzędy, leg. a. czubała, 1999-10-27, det. a. matwiejuk, 2014-12-10. number of specimens examined: 10. cladonia pyxidata (l.) hoffm., deutschl. fl. 2: 121 (1796) this species is distinguished by the corticate and regular scyphose podetia cover with peltate and flat squamules in upper part. podetia are to 2.5 cm high, trumpet-shaped. scyphi are up to 0.6 cm wide. surface of podetia and scyphi covered with an irregular areolate cortex. apothecia are brown, rare, on scyphi margins. for a detailed description of the c. pyxidata see aptroot et al. [21], kowalewska et al. [2]. substances detected by tlc are only fumarprotocetraric acid complex. cladonia pyxidata is morphologically similar to c. monomorpha and c. pocillum, which differs from the podetial surface, rink-like or globose apothecia [2,6,21]. in ne poland, the species was recorded on soil, mainly in well-lit pine forests (12 specimens). one specimen was collected from wood. location of stands of c. pyxidata in ne poland from the herbarium collections of university of bialystok has been presented in fig. 9. this species is rare in north part of poland, but it is common in southern poland [2]. the species is known worldwide [8,18]. it is common in the arctic and temperate zones [15]. specimens examined. ne poland, puszcza knyszyńska landscape park, young forest pine, soil, leg. k. kulikowska, 25.08.992, det. a. matwiejuk, 2014-12-10; wigry national park, studziany las reserve, soil, leg. j. bystrek, a. matwiejuk, 1993-08-06, det. a. matwiejuk, 2014-12-10; biebrzańska kotlina, grzędy, soil, leg. a. czubała, 1999-10-27, det. a. matwiejuk, 2014-12-10; biebrzańska kotlina, grzędy, soil, leg. a. czubała, 199910-27, det. a. matwiejuk, 2014-12-10; biebrzańska kotlina, grzędy, soil, leg. a. czubała, 2001-03-26, det. a. matwiejuk, 2014-12-10; biebrzańska kotlina, grzędy, soil, leg. a. czubała, 1999-10-27, et. a. matwiejuk, 2014-12-10; biebrzańska kotlina, grzędy, soil, leg. a. czubała, 1999-10-27, det. a. matwiejuk, 2014-1210; biebrzańska kotlina, grzędy, soil, leg. a. czubała, 1999-10-27, det. a. matwiejuk, 2014-12-10; klejniki, logging, soil, leg. m. pietryszek, 1999-05-04, det. a. matwiejuk, 2014-12-10; klejniki, logging, soil, leg. m. pietryszek, 1998-07-13, det. a. matwiejuk, 2014-12-10; tyniewicze, slope logging, soil, leg. m. pietryszek, 199904-18, det. a. matwiejuk, 2014-12-10; surroundings of ciechanowiec, forest, wood, leg. s. zabuska, 2002-0717, det. a. matwiejuk, 2014-12-10; forest near rykaczew, soil, leg. a. gosk, 2000-07-11, det. a. matwiejuk, 2014-12-10. number of specimens examined: 13.   białystok podlaskie mazowieckie lubelskie suwalski pk warmińsko mazurskie pk puszczy knyszyńskiej łomżyński pk doliny narwi pk podlaski przełom bugu fig. 9. location of investigation sites of cladonia pyxidata narew bie brz a bug fig. 9 location of investigation sites of cladonia pyxidata. 10 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland discussion and conclusion the taxa from c. pyxidata-chlorophaea group are probably the earliest designated group of chemical taxa [9]. the importance of secondary lichen metabolites to the taxonomy and species discrimination within the c. pyxidata-chlorophaea group has been confirmed based upon simultaneous analyses of dna sequences and morphological and chemical data by stenroos et al. [10]. in ne poland, the chemical variation of the c. pyxidata-chlorophaea group was much neglected, as tlc has not been used in delimitation of species differing in the chemistry. as a great part of herbarium collections of university of bialystok from ne poland was misidentified, i found my research highly needed. except for the samples of c. fimbriata, many studied specimens were misidentified. so far, the lichens from the group c. pyxidata-chlorophaea and allied species has been reported from different regions of poland mostly based on the identification using methods of classical taxonomy [19]. the information about the species recognizable only by secondary substances is rather sparse. in northeastern part of poland, only a few studies have used these laboratory techniques for the identification of c. pyxidata-chlorophaea group so far [2]. altogether, nine species of the c. pyxidatachlorophaea group have been recognized in the examined materials. three taxa (c. asahinae, c. humulis, and c. pocillum) have not been reported up to now from ne poland [2]. cladonia asahinae was found in the mountains. cladonia humilis was known from a few localities along the baltic coast, and c. pocillum from southern poland [2]. cladonia fimbriata is the most common species in poland [2] and ne poland. for comparison, c. grayi is the commonest species in belarus (ca. 40% of the studied specimens) and c. pyxidata is uncommon in belarus, known only from 10 localities [9]. in poland, c. homosekikaica nuno has not reported from c. pyxidata-chlorophaea group. this species was reported in the neighboring countries of poland, in belarus [9] and lithuania [24]. the lichens from the group c. pyxidata-chlorophaea were found in ne poland on soil, in open and sun-exposed sites, and in pine forest. three species (c. chlorophaea, c. fimbriata, c. novochlorophaea) have been reported on bark of trees. cladonia chlorophaea, c. fimbriata, c. grayi, and c. novochlorophaea inhabit wood. only one specimens of c. fimbriata has been reported on rock. similar habitat requirements for many species were reported by kowalewska et al. [2]. the paper presents a number of new localities of species of c. pyxidata-chlorophaea group for ne poland [2]. additionally, a bark of robinia pseudoacacia as a new substrate to c. chlorophaea has been found. the use of tlc method for the identification of lichens collected in this region of poland showed several specimens of rare species: c. conista, c. cryptochlorophaea, c. monomorpha, and c. novochlorophaea. the studies have shown that species c. pyxidata-chlorophaea group differ in their chemical characteristics. in four species of this group, the only chemical component is fumarprotocetraric acid complex (c. chlorophaea, c. fimbriata, c. monomorpha, c. pyxidata). in other species occur fumarprotocetraric acid and other acids. bourgeanic acid is detected in one species (c. conista). gyrophoric acid has been found only in c. grayi. cryptochlorophaeic and paludosic acids have been detected in c. cryptochlorophaea. in c. merochlorophaea, substances detected by tlc include merochlorophaeic and 4-o-methylcryptochlorophaeic acids. homosekikaic and sekikaic acids have been detected in c. merochlorophaea. similar chemical content for many species were reported by kowalewska et al. [2]. presented study confirmed utility of chemical methods in the diagnosis of the species of c. pyxidata-chlorophaea group. 11 of 11© the author(s) 2017 published by polish botanical society acta mycol 51(2):1087 matwiejuk / cladonia pyxidata-chlorophaea group from northeastern poland references 1. ahti t. correlation of the chemical and morphological characters in cladonia chlorophaea and allied lichens. ann bot fenn. 1996;3:380–389. 2. kowalewska a, kukwa m, ostrowska i, jabłońska a, oset m, szok j. the lichens of the cladonia pyxidata-chlorophaea group and allied species in poland. herzogia. 2008;21:61– 78. 3. culberson cf, culberson wl, johnson a. orcinol-type depsides and depsidones in the lichens of the cladonia chlorophaea group (ascomycotina, cladoniaceae). bryologist. 1985;88:380–387. http://dx.doi.org/10.2307/3242681 4. wirth v. die flechten baden-württembergs, teil 1 & 2. stuttgart: ulmer; 1995. 5. brodo im, ahti t. lichens and lichenicolous fungi of the queen charlotte islands, british columbia, canada. can j bot. 1996;74:1147–1180. http://dx.doi.org/10.1139/b96-139 6. james pw. cladonia p. browne. in: smith cw, aptroot a, coppins bj, fletcher a, gilbert ol, james pw, et al., editors. the lichens of great britain and ireland. london: british lichen society; 2009. p. 309–338. 7. holien h, tønsberg t. notes on cladonia asahinae, c. conista and the c. grayi-group in norway. gunneria. 1985;51:1–26. 8. ahti t, stenroos s. cladonia p. browne. in: ahti t, stenroos s, moberg r, editors. nordic lichen flora. vol. 5. cladoniaceae. uppsala: nordic lichen society; 2013. p. 8–87. 9. tsurykau a, golubkov v. the lichens of the cladonia pyxidata-chlorophaea complex in belarus. folia cryptogam est. 2015;52:63–71. http://dx.doi.org/10.12697/fce.2015.52.08 10. stenroos s, hyvönen j, myllys l, thell a, ahti t. phylogeny of the genus cladonia s. lat. (cladoniaceae, ascomycetes) inferred from molecular, morphological, and chemical data. cladistics. 2002;18:237–278. http://dx.doi.org/10.1111/j.1096-0031.2002.tb00151.x 11. kotelko r, piercey-normore md. cladonia pyxidata and c. pocillum; genetic evidence to regard them as conspecific. mycologia. 2010;102:534–545. http://dx.doi.org/10.3852/09-030 12. pino-bodas r, ahti t, stenroos s, martín mp, burgaz ar. cladonia conista and c. humilis (cladoniaceae) are different species. bibl lichenol. 2012;108:161–176. 13. orange a, james pw, white fj. microchemical methods for the identification of lichens. london: british lichen society; 2001. 14. purvis ow, james pw. cladonia hill ex browne (1756). in: purvis ow, coppins bj, hawksworth dl, james pw, moore dm, editors. the lichen flora of great britain and ireland. london: natural history museum publications; 1992. p. 188–210. 15. ahti t. cladoniaceae. bronx, ny: the new york botanical garden; 2000. (flora neotropica monograph; vol 78). 16. ahti t, hammer s. cladonia. in: nash iii th, ryan bd, gries c, bungartz f, editors. lichen flora of the greater sonoran desert region 1. tempe, az: arizona state university; 2000. p. 131–158. 17. archer aw. cladonia chlorophaea sens. lat. in southeastern australia. bryologist. 1983;86:251–253. http://dx.doi.org/10.2307/3242713 18. stenroos s. taxonomy and distribution of the lichen family cladoniaceae in the antarctic and peri-antarctic regions. cryptogamic botany. 1993;3:310–344. 19. fałtynowicz f. the lichens, lichenicolous and allied fungi of poland. an annotated checklist. cracow: w. szafer institute of botany, polish academy of sciences; 2003. 20. osyczka p. a morphometric evaluation of the cladonia chlorophaea group and allied taxa (cladoniaceae, ascomycota). herzogia. 2013;26:49–64 http://dx.doi.org/10.13158/heia.26.1.2013.49 21. aptroot a, sipman hjm, van herk cm. cladonia monomorpha, a neglected cup lichen from europe. lichenologist. 2001;33:271–283. http://dx.doi.org/10.1006/lich.2001.0332 22. kowalewska a, kukwa m. new records of cladonia monomorpha (cladoniaceae, lichenised ascomycota) from europe. herzogia. 2004;17:103–105. 23. øvstedal do, lewis smith ri. lichens of antarctica and south georgia. a guide to their identification and ecology. cambridge: cambridge university press; 2001. 24. motiejūnaitė j. lapiškosios ir krūmiškosios kerpės. vilnius: botanikos insitutas; 2002. [lietuvos grybai; vol 13(1)]. http://dx.doi.org/10.2307/3242681 http://dx.doi.org/10.1139/b96-139 http://dx.doi.org/10.12697/fce.2015.52.08 http://dx.doi.org/10.1111/j.1096-0031.2002.tb00151.x http://dx.doi.org/10.3852/09-030 http://dx.doi.org/10.2307/3242713 http://dx.doi.org/10.13158/heia.26.1.2013.49 http://dx.doi.org/10.1006/lich.2001.0332 abstract introduction material and methods results cladonia chlorophaea (flörke ex sommerf.) spreng., syst. veg. 4: 273(1827) cladonia conista robbins ex a. evans, trans. connecticut acad. arts 30: 472 (1930) cladonia cryptochlorophaea asahina, j. jap. bot. 16: 711 (1940) cladonia fimbriata (l.) fr., lichenogr. eur. ref.: 222 (1831) cladonia grayi g. merr. ex sandst., clad. exs. no. 1847 (1929) cladonia merochlorophaea asahina, j. jap. bot. 16: 713 (1940) cladonia monomorpha aptroot, sipman & van herk, lichenologist 33: 273 (2001) cladonia novochlorophaea (sipman) brodo & ahti, canad. j. bot. 74: 1167 (1996) cladonia pyxidata (l.) hoffm., deutschl. fl. 2: 121 (1796) discussion and conclusion references 2017-01-16t18:18:36+0000 piotr otręba new records of rare lichenicolous and lichen-forming fungi from volcanic rocks in sw poland 1 of 8published by polish botanical society acta mycologica short communication new records of rare lichenicolous and lichen-forming fungi from volcanic rocks in sw poland katarzyna szczepańska* department of botany and plant ecology, wrocław university of environmental and life sciences, pl. grunwaldzki 24a, 50-363 wrocław, poland * email: katarzyna.szczepanska@up.wroc.pl abstract records of two lichenicolous and nine lichen-forming fungi found in the southwestern part of poland are presented. all of the reported species are very rare and they have only a few scattered localities in the country. one of them, lecanora pannonica, is reported for the second time from poland. additionally, the new, contemporary records of cercidospora macrospora, rhizocarpon disporum, r. viridiatrum and stereocaulon pileatum in lower silesia were noted. these species were known only from historical collections in the study area. furthermore, lecidea fuscoatra has been found a new host for sagediopsis barbara. all of the localities of recorded species were found on natural outcrops of basalt rocks. keywords ascomycota; biodiversity; distribution; basalt rocks this issue of acta mycologica is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. introduction the southwestern part of the poland has the biggest number of basalt rock formations in the country. the rocks in the region are a part of a large central european petrographic province reaching from moravian ostrava and opavan silesia, through opole silesia, the sudetes, upper lusatia, saxony and bohemian central uplands to the rhine valley and the borders of the netherlands [1]. within polish borders the biggest clusters of the rocks are located near lubań, zgorzelec, between lwówek śląski and jawor, and between gryfów śląski and świeradów. these clusters are relics of old volcanic fields, of which every group could have had up to a hundred of small volcanoes [2]. basalt volcanism is connected with the youngest tectonic movements (in the cenozoic era) in the sudetes and their vicinity, which were mostly vertical movements. rocks created in this way exist today mostly as various forms of lava veins – volcanic plugs, flood basalt, volcanic cones and calderas [1]. basalts (volcanic rocks made of plagioclase, pyroxene and olivine) are very hard volcanic rocks with absorptivity and, therefore, they are resistant to repeated processes of freezing and thawing. because of that they often form denudation monadnocks – inselbergs [3]. it doi: 10.5586/am.1056 publication history received: 2015-02-12 accepted: 2015-07-02 published: 2015-08-05 handling editor maria rudawska, institute of dendrology of the polish academy of sciences, poland funding the study was funded by department of botany and plant ecology, wrocław university of environmental and life sciences. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation szczepańska k. new records of rare lichenicolous and lichenforming fungi from volcanic rocks in sw poland. acta mycol. 2015;50(1):1056. http://dx.doi. org/10.5586/am.1056 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:katarzyna.szczepanska%40up.wroc.pl?subject=new%20records%20of%20rare%20lichenicolous%20and%20lichen-forming%20fungi%20from%20volcanic%20rocks%20in%20sw%20poland http://dx.doi.org/10.5586/am.1056 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1056 http://dx.doi.org/10.5586/am.1056 2 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1056 szczepańska / new records of lichens and lichenicolous fungi in poland is estimated that there are about 300 various basalt rock formations in lower silesia [2], most of which were in the past centuries used for mining. many of these formations are extremely valuable for scientific, didactic, aesthetic and natural reasons and are often protected as landmarks [4]. when the rocks decompose due to environmental factors they create fertile soil and decomposed rocks rich in mineral compounds. for that reason many rare plants and plant communities can be found in their vicinity [5–9]. volcanic rocks, including basalt rocks, often foster rare species lichens and lichenicolous fungi because of unique habitat conditions [10–16]. the current paper includes data on 11 species of lichens and lichenicolous fungi occurring in sw poland. these species are mostly known from historical collections in the study area and they are currently very rare in the country. all of the presented species are saxicolous and had been discovered on natural outcrops of volcanic rock formations. additionally, their ecology and distribution in poland and in central europe are briefly characterized. material and methods material included in the present paper was collected by the author in 2012–2014, on some natural outcrops of volcanic rocks, especially on basalt rocks occurring in sw poland. material was determined using standard methods. identification of sterile, crustose specimens was supported by tlc analyses of secondary metabolites, according to the methods of orange et al. [17]. the distribution of the taxa examined are given in the atpol grid square system [18], modified by cieśliński and fałtynowicz [19]. the herbarium material is housed in the private herbarium of the author (hb. szczepańska). in the text the asterisk (“*”) indicates a lichenicolous fungus. results and discussion buellia badia (fr.) a. massal. this species is characterized by subsquamulose to squamulose, brown thallus. it is considered as a saxicolous, however it often grows as a parasite over various lichens, such as melanelia and xanthoparmelia species [20]. in poland it is known from the pojezierza bałtyckie lakelands, nizina środkowopolska lowland and wysoczyzna podlaska height [21]. in sw poland it was noted only on the few localities in the sudety mts [22–24]. on the new location, some of the young thalli of b. badia, were growing on the thallus of xanthoparmelia loxodes (nyl.) o. blanco et al. in central europe, b. badia has been found in austria [25], the czech republic [26] and germany [27]. specimen examined. eb-40: pogórze zachodniosudeckie foothills, pogórze kaczawskie foothills, ostrzyca proboszczowicka mt, alt. 450 m, on basalt rocks, 26 september 2012, leg. k. szczepańska 929 (hb. szczepańska). caloplaca subpallida h. magn. this species belongs to the caloplaca ferruginea group [28] which includes taxa characterized by whitish to blackish thalli and ferrugineous apothecia. it was reported for the first time, as a new to poland, in the sudety mts, including przedgórze sudeckie foreland and pogórze zachodniosudeckie foothills [16]. it grows only on mineralrich, siliceous rocky substrata, especially on the basic and altered ultrabasic rocks, i.e., basalt, greenstone and serpentinite. in central europe, c. subpallida has been found in austria [25], the czech republic [26] and germany [27]. 3 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1056 szczepańska / new records of lichens and lichenicolous fungi in poland specimen examined. eb-64: przedgórze sudeckie foreland, wzgórza strzegomskie hills, góra krzyżowa mt, alt. 360 m, on basalt rocks, 25 may 2013, leg. k. szczepańska 949 (hb. szczepańska). *cercidospora macrospora (uloth) hafellner & nav.-ros. a rare lichenicolous fungus in poland, reported from only a few localities till now. it was noted in the pomorze gdańskie pomerania and in the xix century in the sudety mts [29]. cercidospora macrospora was recorded in poland exclusively on the thallus of the lecanora muralis (schreb.) rabenh., however this taxon is also known as growing on different lecanora species, occurring on siliceous, as well as on calcareous rocks [30]. in contrary to other, known from poland, species – c. epipolytropa (muss) arnold, c. macrospora does not show a preference to develop the ascomata in the apothecial discs and they are equally frequent in host thallus and apothecia [31]. in central europe, c. macrospora has been found in austria [31], the czech republic [32], germany and slovakia [31]. host: lecanora muralis (schreb.) rabenh. (apothecia and thallus). specimen examined. eb-64: przedgórze sudeckie foreland, wzgórza strzegomskie hills, góra krzyżowa mt, alt. 360 m, on lecanora muralis growing on basalt rocks, 19 may 2013, leg. k. szczepańska 982 (hb. szczepańska). lecanora subaurea zahlbr. lecanora subaurea is listed on the red list of the lichens in poland [33], in dd category (data deficient). as a habitat it prefers heavy metal rich lime-free silicate rocks, including iron-rich basalts [27]. due to similar, bright yellow color of the thallus, as well as secondary chemistry and habitat, l. subaurea can be confused with l. epanora (ach.) ach. however, in contrary to l. epanora, soredia of l. subaurea arising on the margins of areoles, not on their upper surface. in the country this species has been noted mainly in the southern part of the country, in the karpaty zachodnie mts [21,34]. additionally it has two localities in the sudety mts, in karkonosze mts [35,36] and rudawy janowickie mts [37]. some of the specimens noted on the słupiec rock were growing as a parasite, on the aspicilia simoënsis räsänen thallus. the specimens examined by tlc method contained typical substances mentioned in literature [27,38], i.e., pannarin (major), rhizocarpic acid (major) and zeorin (major). in central europe, l. subaurea has been found in austria [25], the czech republic [26] and germany [27]. specimens examined. ea-56: pogórze izerskie foothills, ciasnota mt, stożek perkuna rock near grabiszyce village, alt. 401 m, on basalt rock, 21 june 2014, leg. k. szczepańska 1001 (hb. szczepańska). ea-57: pogórze izerskie foothills, słupiec rock near giebułtów village, alt. 746 m, on basalt rock, 22 june 2013, leg. k. szczepańska 962, 984 (hb. szczepańska). lecanora pannonica szatala this species was reported only once from northern poland, in the pojezierze iławskie lackeland and nizina staropruska lowland [39], from three localities. lecanora pannonica usually grows on anthropogenic substrates [38] and all of its previously known records from poland, come from old brick made buildings. the new locality of this species in country, for the first time was recorded on natural rocky substrata. this species was observed on vertical surfaces of well lighted basalt rocks, however there were found only sterile thalli. lecanora pannonica can be confused with tephromela grumosa (pers.) hafellner & cl. roux, which is also usually sterile lichen, produce blue-grey, k+ yellow soredia. the features which allow to distinguish l. pannonica from t. grumosa, are distinctly bullate-areolate thallus, discrete, rounded and 4 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1056 szczepańska / new records of lichens and lichenicolous fungi in poland not confluent soralia as well as chemistry. the specimens were analyzed using tlc method. the thallus contains atranorin (major), roccellic acid (major) and gangaleoidin (trace). in central europe, l. pannonica has been found in austria [25], germany [27] and slovakia [40]. specimen examined. eb-64: przedgórze sudeckie foreland, wzgórza strzegomskie hills, góra krzyżowa mt, alt. 360 m, on basalt rocks, 19 may 2013, leg. k. szczepańska 955 (hb. szczepańska). lecidella scabra (taylor) hertel & leuckert a lichen species listed on the red list of the lichens in poland [33], in nt category (near threatened). due to similar c+ orange reaction of soredia, l. scabra may be confused with rinodina aspersa (borrer) j.r. laundon. however, r. aspersa differs from l. scabra in secondary chemistry and because of possessing clearly areolate thallus with discrete, not confluent, circular soralia. lecidella scabra has scattered localities in poland, situated in the pojezierza bałtyckie lakelands, wysoczyzna podlaska height, wyżyna środkowomałopolska upland and karpaty zachodnie mts [21,34]. in sw poland, l. scabra was noted only in the sudety mts [41]. most of the specimens found on the new locality were sterile and in order to proper identification of the specimens, tlc analysis was performed. thallus of the species contains atranorin (major) and xanthones: thuringione (minor) and arthothelin (major). additionally there was found some unidentified substance (rf class in solvent c – 35, spot color after acid and heating – colorless, uv after heating – bright yellowish-green). in central europe, l. scabra has been found in austria [25], the czech republic [26], germany [27] and hungary [42]. specimen examined. eb-64: przedgórze sudeckie foreland, wzgórza strzegomskie hills, góra św. jerzego mt, alt. 360 m, on basalt rocks, 25 may 2013, leg. k. szczepańska 954 (hb. szczepańska). rhizocarpon disporum (nägeli ex hepp) müll. arg. this species has only a few scattered localities in poland. it has been noted mainly in the mountain areas in the southern part of the country [21,34] and additionally in the pojezierze bałtyckie lakelands [43]. in sw part of the country it was observed only in xix century in the sudety mts [22,23,44–46]. rhizocarpon disporum is characterized by the 1-spored asci, containing very large (40–70 × 18–30 μm), dark and strongly muriform ascospore [27]. in central europe, r. disporum was noted in austria [25], the czech republic [26], germany [27] and hungary [42]. specimen examined. eb-64: przedgórze sudeckie foreland, wzgórza strzegomskie hills, góra krzyżowa mt, alt. 360 m, on basalt rocks, 4 october 2013, leg. k. szczepańska 971 (hb. szczepańska). rhizocarpon viridiatrum (wulfen) körb. rhizocarpon viridiatrum is considered as a rare species in poland and is listed on the red list of the lichens in poland [33], in vu category (vulnerable). it was recorded in the pojezierza bałtyckie lakelands and karpaty zachodnie mts [21,34]. in sw poland r. viridiatrum was noted only in historical papers, in the sudety mts [22,23,45,46]. young thalli of this species are known as initially parasitic on other saxicolous, crustose lichens, especially on aspicilia caesiocinerea (malbr.) arnold [38]. on the new locality it was found growing on the aspicilia cinerea (l.) körb. thallus, in light, exposed but rather warm places. 5 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1056 szczepańska / new records of lichens and lichenicolous fungi in poland in central europe, r. viridiatrum has been found in austria [25], the czech republic [26], germany [27] and hungary [42]. specimen examined. eb-64: przedgórze sudeckie foreland, wzgórza strzegomskie hills, góra świętego jerzego mt, alt. 354 m, on aspicilia cinerea on basalt rocks, 19 may 2013, leg. k. szczepańska 957 (hb. szczepańska). stereocaulon pileatum ach. this is a mountain species, listed on the red list of the lichens in poland [33], in en category (endangered). it was noted only in the southern part of the country, on the few localities, in the karpaty zachodnie mts [21,34] and sudety mts [23,45,47,48]. this species is characterized by wart-like phyllocladia and distinctly globose soralia, located at the ends of small pseudopodetia. the appearance and position of soralia distinguishes this species from the other taxon with small thallus – s. nanodes tuck. till now, s. pileatum has been known as growing only on sandstone and granite rocks and this is the first record of this species noted on the basalts. in central europe, s. pileatum has been found in austria [25], the czech republic [26] and germany [27]. specimens examined. ea-56: pogórze izerskie foothills, ciasnota mt, stożek perkuna rock near grabiszyce village, alt. 401 m, on basalt rock, 21 june 2014, leg. k. szczepańska 1000 (hb. szczepańska). ea-57: pogórze izerskie foothills, słupiec rock near giebułtów village, alt. 746 m, on basalt rock, 22 june 2013, leg. k. szczepańska 958 (hb. szczepańska). *sagediopsis barbara (th. fr.) r. sant. & triebel a rare lichenicolous fungus in poland, reported only from the sudety mts [29]. it is known as a parasite restricted to porpidia rugosa (taylor) coppins & fryday [49], however, some others lichens, like lecanora and lecidea genera, are mentioned as a hosts of this taxon [50]. on the new localities s. barbara was noted on the thallus of the lecidea fuscoatra (l.) ach. this species differs from similar s. aquatica (stein) triebel mainly in the larger size of perithecia and ascospores. characteristics of specimens: ascomata perithecioid, 380–450 μm diam., globose, black, partly immersed in the host thallus, ascomata wall dark brown to black, perithecium wall distinctly delimited from the host tissue, interascal filaments visible usually only in early stages in development, asci 8-spored, ascospores hyaline, narrowly fusiform with rounded ends and with only transverse septa, spores (25.5–)30.0– 38.0(–41.0) × (2.0–)3.0–3.5(–4.0) μm. in central europe, s. barbara has been found in austria [51] and the czech republic [32]. host: lecidea fuscoatra (l.) ach. (thallus). specimens examined. ea-56: pogórze izerskie foothills, ciasnota mt, stożek perkuna rock near grabiszyce village, alt. 401 m, on lecidea fuscoatra growing on basalt rock, 21 june 2014, leg. k. szczepańska 1010 (hb. szczepańska). ea-57: pogórze izerskie foothills, słupiec rock near giebułtów village, alt. 746 m, on lecidea fuscoatra growing on basalt rock, 22 june 2013, leg. k. szczepańska 1011 (hb. szczepańska). tephromela grumosa (pers.) hafellner & cl. roux this species is not very frequent in poland, especially in the sw part of the country. it is known from the pojezierza bałtyckie lakelands, wysoczyzna podlaska height, wyżyna środkowomałopolska upland [21] and sudety mts [22,23,41,52,53]. at the new localities only sterile thalli of this species were found. tephromela grumosa can be confused with other saxicolous and usually sterile taxa – lecanora pannonica (see 6 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1056 szczepańska / new records of lichens and lichenicolous fungi in poland comments on l. pannonica). secondary metabolites detected by tlc in analyzed specimens were atranorin (major) and protolichesterinic acid (major). there were not find α-collatolic acid, which is sometimes mentioned as a substance present in t. grumosa thallus [27]. in central europe, t. grumosa has been found in austria [25], the czech republic [26] and germany [27]. specimens examined. eb-40: pogórze zachodniosudeckie foothills, pogórze kaczawskie foothills, ostrzyca proboszczowicka mt, alt. 450 m, on basalt rocks, 26 september 2012, leg. k. szczepańska 934 (hb. szczepańska). ea-57: pogórze izerskie foothills, słupiec rock near giebułtów village, alt. 746 m, on basalt rocks, 22 june 2013, leg. k. szczepańska (hb. szczepańska). eb-64: przedgórze sudeckie foreland, wzgórza strzegomskie hills, góra krzyżowa mt, alt. 360 m, on basalt rocks, 25 may 2013, leg. k. szczepańska (hb. szczepańska). references 1. birkenmajer k. lower silesian basalts as monuments of inanimate nature. ochrona przyrody. 1967;32:225–276. 2. krzemińska e, awdankiewicz m. geological history of volcanic activity in the territory of poland. kosmos. 2011;60(3–4):261–275. 3. adam a. die schichtstufenlandschaft der vorberge des bober-katzbach-gebirges und des nordöstlichen teils des lsergebirges. przyroda sudetów. 2004;7:175–190. 4. grocholski a, jerzmański j. paleovolcanic occurrence in the lower silesia, in the light of nature protection. ochrona przyrody. 1975;40:291–349. 5. fabiszewski j. o roślinności ciepłolubnej na bazaltach góry krzyżowej koło strzegomia. chrońmy przyr ojcz. 1963;5:16–21. 6. anioł-kwiatkowska j, świerkosz k. flora and plant communities of ostrzyca proboszczowicka hill. acta univ wratislav prace bot. 1992;48:45–117. 7. kwiatkowski p. vegetation of the planned reserve “góra zamkowa” near wleń (western sudety). acta univ wratislav prace bot. 1994;40:95–113. 8. szczęśniak e. vegetation of the projected “krzyżowa góra near strzegom” reserve (lower silesia). ochrona przyrody. 1998;55:61–75. 9. pędziwiatr a. bazaltowa flora. przyroda polska. 2013;2:24–25. 10. kossowska m. pertusaria lactescens (lichenized ascomycota, pertusariales), a lichen species new to central europe. pol bot j. 2000;53(1):69–70. 11. kossowska m. notes on leptogium and dermatocarpon species (lichenized ascomycota) from a basalt outcrop in mały śnieżny kocioł cirque (karkonosze mts, poland). pol bot j. 2008;53(1):95–96. 12. kossowska m. new and interesting lichenicolous fungi of the karkonosze mountains, sw poland. herzogia. 2008;21:229–232. 13. kossowska m. new, rare and noteworthy lichens in the giant mountains. biologia. 2011;66(5):755–761. http://dx.doi.org/10.2478/s11756-011-0084-4 14. szczepańska k. lichens of the “kruczy kamień” nature reserve in góry kamienne mountains (central sudety mountains). in: lipnicki l, editor. lichen protection – protected lichen species. gorzów wlkp: sonar literacki; 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(monographiae botanicae; vol 26). abstract introduction material and methods results and discussion buellia badia (fr.) a. massal. caloplaca subpallida h. magn. *cercidospora macrospora (uloth) hafellner & nav.-ros. lecanora subaurea zahlbr. lecanora pannonica szatala lecidella scabra (taylor) hertel & leuckert rhizocarpon disporum (nägeli ex hepp) müll. arg. rhizocarpon viridiatrum (wulfen) körb. stereocaulon pileatum ach. *sagediopsis barbara (th. fr.) r. sant. & triebel tephromela grumosa (pers.) hafellner & cl. roux references 2015-08-02t16:01:41+0100 piotr otręba can macrofungal biodiversity predict forest status and dynamics? a view from south european mediterranean forests (italy) acta mycologica article id: 567 doi: 10.5586/am.567 publication history received: 2020-09-27 accepted: 2021-03-20 published: 2021-07-29 handling editor andrzej szczepkowski; warsaw university of life sciences – sggw, poland; https://orcid.org/0000-00029778-9567 authors’ contributions ea: conceptualization, investigation, methodology, writing – original draft, writing – review and editing, data analysis; af: conceptualization, methodology, review and editing funding this study was carried out in the framework of a phd project in applied botany in agriculture and the environment, university of genoa, italy. ea was supported by miur (ministry of education, universities and research – italy) doctoral fellowship (2012–2014). competing interests no competing interests have been declared. copyright notice © the author(s) 2021. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. original research paper in ecology of fungi can macrofungal biodiversity predict forest status and dynamics? a view from south european mediterranean forests (italy) elia ambrosio1*, alan feest 2,3 1department of earth, environment and life science (distav), university of genoa, corso europa 26, genoa, 16132, italy 2faculty of engineering, queen’s building, university of bristol, university walk, bristol, bs8 1tr, united kingdom 3ecosulis harwell innovation centre, building 173, curie avenue, oxford, o11 oqg, united kingdom * to whom correspondence should be addressed. email: elia.ambrosio.10@gmail.com abstract fungi are among the most important organisms on earth, and they are essential components of terrestrial ecosystems. eir reproductive structures are strictly dependent and affected by environmental conditions, and community dynamics over time and space may be indirect indicators of the health status of forests. we combined macrofungal biodiversity indices in eight mediterranean forests in italy and surveyed 160 plots by standardized surveys, to evaluate the role of macrofungi as early predictors of change in the forest structure. e results show that indices of fungal diversity are influenced by geographic and floristic conditions, and interand intra-annual temperature and rainfall fluctuations affect the formation of fungal fruiting bodies. ese findings suggest that environmental changes could be reflected by macrofungi, and conservation initiatives should consider the pivotal role that fungi play in biodiversity monitoring. keywords environmental impact; macrofungi; biodiversity indices; standardized surveys; mediterranean forests 1. introduction e health status of forests on a global scale has changed considerably because of the acceleration of climate change and collapse of ecosystems (curtis et al., 2018; omas et al., 2004). e protection and restoration of forests is crucial for the mitigation of global climate change and biodiversity conservation (chiarucci & piovesan, 2019). issues arise when considering how to monitor forest dynamics, measure the health status of forest ecosystems, and in determine the extent of human impact. biodiversity has been viewed as a quantitative characteristic that can be assessed or measured by a number of direct indices reflecting the overall holistic characteristics of biodiversity (feest, 2013), here described as “biodiversity quality” (feest, 2006). a recent study by ambrosio et al. (2018) and feest (2013) showed that relevant information regarding biodiversity may be extrapolated through organisms that are indicators of ecosystem functions and conditions. is aspect is particularly important given the need for the development of methodologies that would reduce the time and costs involved in biological conservation and monitoring actions (united nations, 2017). in this sense, fungal biodiversity can potentially provide an indication of ecosystem changes because it is closely related to overall site biodiversity and expresses the heterogeneity of a habitat (boddy et al., 2014; büntgen et al., 2011; egli, 2011; kauserud et al., 2008, 2010; kotiranta & niemelä, 1993). several studies have acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 1 https://doi.org/10.5586/am.567 https://orcid.org/0000-0002-9778-9567 https://orcid.org/0000-0002-9778-9567 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0003-1299-2513 mailto:elia.ambrosio.10@gmail.com ambrosio and feest / macrofungi and forest health status confirmed that changes in fungal communities may reflect the degree of forest degradation (egli, 2011; stursova et al., 2014). hence, monitoring the composition and dynamics of the fungal community over time and space may represent a useful tool for evaluating the health status of forests and establishing conservation priorities in different locations (egli, 2011; feest, 2013; halme & kotiaho, 2012). fungi are among the most important organisms on earth and are essential components of terrestrial ecosystems because of their functional roles and high species richness and diversity (blackwell, 2011; hawksworth, 2001; mueller & schmit, 2007). ey profoundly affect forest ecosystem functions by driving the carbon cycle, decomposing organic matter, mediating nutrient and water uptake, protecting roots from fungal pathogens, and maintaining soil structure and forest food webs (heilmann-clausen et al., 2014; tedersoo et al., 2014). moreover, the fungal reproductive (and ephemeral) structures (viz. fruiting bodies/sporocarps) are strictly dependent and affected by climatic (e.g., light, microclimate, rainfall) and edaphic conditions (e.g., ph, nutritional factors, organic matter) (boddy et al., 2014). in vitro growth experiments have shown that climate variables (e.g., rising temperatures) influence fungal physiology and growth (boddy et al., 2014); hence, their presence/absence, or community variation over time and space may be used as indirect indicators of climate change and predictors of forest health status and dynamics. due to the key role played by fungi in detecting early changes in habitats and environments, e.g., nitrogen deposition as in baar and kuyper (1998), feest et al. (2014), and mohan et al. (2014), as well as their importance in ecological and conservation studies, the macrofungal biodiversity quality of eight mediterranean forests located in italy was investigated using a standardized survey approach (feest, 2006). e goals of this study were: (i) to evaluate the influence and relationship of forest types and management on macrofungal biodiversity quality indices; (ii) to identify possible macrofungal species indicators of mediterranean forests; and (iii) to assess macrofungal biodiversity quality over the years. we postulated the following hypotheses: 1. different mediterranean forests, which differ in vegetation type and management, have statistically different macrofungal biodiversity quality indices. 2. a site surveyed over consecutive years will have statistically different macrofungal biodiversity quality indices as a result of abiotic factors (e.g., climate variations). in this study, biodiversity quality is regarded as the quality of a site that can be extrapolated from a number of measured characteristics of the population studied (here macrofungi) that are present at the time of sampling (ambrosio et al., 2018; feest, 2006). 2. material and methods 2.1. study sites eight sites were selected from northern to southern italy (including islands) along a latitudinal gradient (figure 1). ese forest types are representative of mediterranean (italian) vegetation (blasi et al., 2007; mariotti, 2008). specifically, site 1 (s1 in figures and tables) is a silver fir plantation (abies alba mill.) located in sardinia; sites 2 (s2) and 8 (s8) are both chestnut coppices, dominated by castanea sativa mill. [lower frequency of other woody species occurs at these sites, such as sorbus torminalis (l.) crantz and populus tremula l.], which are located in liguria (s2) and campania (s8), respectively. both sites (s2 and s8) are subject to human intervention (i.e., by cutting and thinning) to clear the undergrowth vegetation and facilitate the collection of chestnuts. sites 3 (s3) and 5 (s5) are characterized by deciduous oak (quercus cerris l.) woods and are managed as high priority forests consisting of large, tall, and mature trees with a closed canopy. a lower frequency of s. torminalis and p. tremula occurred at these sites. ey are located in liguria (s3) and tuscany (s5), respectively. site 4 (s4), located in liguria, is a beech forest, which acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 2 ambrosio and feest / macrofungi and forest health status figure 1 location of study sites. is dominated by fagus sylvatica l. (followed by s. torminalis and p. tremula at lower frequencies), and is also managed as a high priority forest. finally, sites 6 (s6) and 7 (s7) are evergreen oak (quercus ilex l.) woods (ascribed to the mediterranean maquis landscape), located in tuscany (s6) and campania (s7), respectively. all the sites are open to public for picking edible mushrooms under regional and national rules, except s2, s3, and s8, where the access is restricted and picking of edible mushrooms is not allowed because they are located on private property. only site 1 can be described as a monoculture. more details on the characteristics of the sites are summarized in table 1. 2.2. survey procedure all study sites were surveyed using the same standardized procedure (feest, 2006). surveys were performed during the period of maximum sporocarps fruiting and development (in fall, mainly early october in 2013) in 20 circular (4-m radius) plots at each study site along line transects. each plot was placed 20 m from the preceding plot. e total sample area was approximately 1,000 m2 per site. sites 2–4 were surveyed over three consecutive years (2012–2014). mycological observations were focused on epigeous macrofungi with a determinate shape that were visible to the naked eye, mostly sporocarps (kirk et al., 2008). macrofungi were studied both qualitatively (identification of species) and quantitatively (number of sporocarps/fruiting bodies per species). taxonomical identification was performed by analyzing the macroand microscopic characteristics of the collected specimens. e systematic classification followed hibbett et al. (2014) and he et al. (2019). nomenclature was used in accordance with acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 3 a m b ro sio an d feest / m acro fu n g ian d fo rest h ealth statu s table 1 characteristics of study sites. site code locality coordinates altitude (ma.s.l.) vegetation forest type geology mean rainfall(mm) mean temperature (°c) data references s1 sardegna – loc. tempio pausania (ot) 40°51′7″ n, 9°10′13″ e 1,262 silver fir – abies alba unmanaged leucogranites rocks 1,300 10 ambrosio et al., 2015 s2 liguria – loc. badani (sv) 44°27′56″ n, 8°28′44″ e 430 chestnut coppice – castanea sativa managed calceschist rock 912 12 ambrosio et al., 2018 s3 liguria – loc. la maddalena (sv) 44°30′14″ n, 8°29′17″ e 360 deciduous oak wood – quercus cerris unmanaged serpentine schists 912 12 ambrosio et al., 2018 s4 liguria – loc. veirera (sv) 44°27′3″ n, 8°32′42″ e 1,000 beech forest – fagus sylvatica unmanaged calceschist rock 912 12 ambrosio et al., 2018 s5 toscana – loc. basciano (si) 43°22′36.1″ n, 11°17′24.9″ e 328 deciduous oak wood – quercus cerris unmanaged calcareous soil 777 13.4 unpublished data s6 toscana – loc. montegabbro (si) 43°23′53.7″ n, 11°02′43.7″ e 297 evergreen oak wood – quercus ilex unmanaged calcareous soil 777 13.4 unpublished data s7 campania – loc. san felice (ce) 41°14′15.9″ n, 13°58′06.8″ e 210 evergreen oak wood – quercus ilex unmanaged calcareous soil 900.8 16.1 unpublished data s8 campania – loc. roccamonfina (ce) 41°18′09.5″ n, 13°59′12.0″ e 612 chestnut coppice – castanea sativa managed vulcanic soil 925.9 15.2 unpublished data a cta m ycologica / 2021 / vo lu m e 56 / a rticle 567 pu b lish er:po lish b o tan icalso ciety 4 ambrosio and feest / macrofungi and forest health status the index fungorum (http://www.indexfungorum.org/), cbs (https://wi.knaw.nl/), and ima (https://www.mycobank.org/) databases. 2.3. data sets full datasets are shown in appendix s1–appendix s8. to calculate a set of biodiversity indices and statistics (detailed in the next paragraph) for each study site, we defined a dataset as: the total number of fungal species and respective sporocarps/fruiting body abundance. datasets refer to both published and unpublished data (see table 1) collected by following the same standardized sampling procedure (feest, 2006). 2.4. data analyses a set of biodiversity indices (magurran, 2004), based on the number of individuals (sporocarps/fruiting bodies) collected per study site that were present at the time of sampling, was computed to extrapolate the biodiversity quality (feest, 2006; feest et al., 2010) of each selected site, as follows: • species richness (sr) is the number of species recorded. e sr expresses alpha diversity and is the simplest measure of biodiversity, albeit with low information value. • population abundance (pa): the number of individuals (sporocarps) for each collected species. • population density (pd): the total number of individuals recorded in the standardized sample (pd/m2). • shannon index (h′) was used to combine the effects of sr and evenness. is index assumes that individuals are randomly sampled, and that all species are represented in the sample. h′depends on the total number of species and their occurrence. • pielou evenness index (e) was calculated to measure the departure of the observed pattern from the expected pattern in a uniform assemblage of equally abundant species. • simpson’s index (d) was computed to calculate the evenness of the population per species. is index is weighted by the abundance of the most common species; when d increases, diversity decreases. • berger–parker index (d) expresses the proportional abundance of the most abundant species. • nonparametric estimators were used to estimate the sr and to check whether data collected allowed observation of a notable number of species. specifically, the chao estimator is the simplest method to extrapolate the predicted number of species in an assemblage, and is based on the number of singleton and doubleton species (chao, 1984; magurran, 2004). e first-order jackknife estimator is based on the number of “unique species,” as well as the exact number of species that occur in the sampling units and duplicates, respectively (heltshe & forrester, 1983; magurran, 2004). finally, the bootstrap estimator for sr was based on the frequency distribution of the species found in the sample (magurran, 2004; smith & van belle, 1984). ese estimators indicate the degree of the sampling efficiency. • rarefaction curves were chosen because they illustrate the rate at which new species are found unless sampling has been exhaustive. e curve generally rises very quickly at first and then flattens once a reasonable number of individuals have been considered (colwell et al., 2012; gotelli & colwell, 2001; magurran, 2004). in this study, the number of species was plotted as a function of the accumulated number of individuals. we used the function “rarefy” in the vegan package version 2.5-7 (oksanen et al., 2013) to plot the curves with the corresponding iterative error around the mean. rarefaction generates the expected number of species in a small collection of individuals drawn randomly from the entire pool of individuals. in this study, “individuals” refer to the sporocarps counted and “sample” to plots in each study site. acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 5 http://www.indexfungorum.org/ https://wi.knaw.nl/ https://www.mycobank.org/ ambrosio and feest / macrofungi and forest health status • species conservation value index (scvi) (feest, 2006; feest et al., 2010) was calculated as the mean commonness/rarity of each species recorded according to national references (boccardo et al., 2008; onofri et al., 2005). accordingly, all species were given a numerical value, as follows: 1 – species occurring in more than 85%–100% of the italian territory; 2 – species recorded on 65%–85%; 3 – species recorded on 45%–65%; 4 – species recorded on 25%–45%; 5 – species recorded on less than 25% of the italian territory. • biomass index (bi) was estimated using a nondestructive method (tóth & feest, 2007). biomass is directly related to the size of fruiting bodies calculated from the radius of the cap. e bi was computed from the biometrics of the species as dry weight (feest, 2006). • indicator species analysis (isa) (de cáceres et al., 2010) was used to identify macrofungal indicator species for the selected study sites. • hierarchical cluster analysis (h-ca) using the bray–curtis dissimilarity index and unweighted pair group method with arithmetic mean upgma (legendre & legendre, 2012) was carried out to discern the degree of similarity between the selected study sites. • correspondence analysis (ca) was performed to determine the distribution of macrofungal communities without the constraint of biotic and abiotic variables. e variables are represented by species and cases by plots (legendre & legendre, 2012). • h-ca and ca were both computed with sr, presence/absence, and shannon index obtained in each surveyed plot (160 in total) for all of the selected sites. • e effect of climatic factors (mean temperatures and rainfall) on macrofungal biodiversity indices (sr, pa, h′, and bi) was tested using the multivariate technique of principal correspondence analysis (pca) (everitt & hothorn, 2006). since the environmental variables were expressed at different measurement scales, pca was computed on the correlation matrix. vector fitting was used to identify significant variables on the ordination axes (pca1 and pca2). all statistical analyses were computed using vegan and biodiversityr packages in the r statistical environment (version 4.0.0; 2020). 2.5. climate data climate data were extracted using the arpal database (https://www.arpal.liguria.it/). specifically, we selected daily data on mean temperature (°c) and mean rainfall (mm) for sites 2, 3 and 4 (from loc. sassello weather station), where we conducted field surveys for three consecutive years (2012, 2013, 2014). 3. results overall, mycological surveys were performed in 160 plots for a total sampled area of 8,000 m2. appendix s1–appendix s8 list the recorded number of species and sporocarp (mainly basidiomata) abundance collected at each site and plot. a total of 153 macrofungal species (species richness) were observed in the eight study sites (in 2013): 59 species in the beech forest (s4), 52 species in the silver fir site (s1), 45 and 23 species in the chestnut coppices (s2 and s8, respectively), 35 and 33 species in the evergreen oak woods (s7 and s5), and 34 and 33 species in the deciduous oak woods (s3 and s6, respectively) (see sr column in table 2). e highest value of population abundance (pa in table 2) and density (pd) were obtained in the evergreen oak woods (s5; pa = 761, pd = 0.761 m−2), while the lowest was obtained in the beech forest (s4; pa = 162, pd = 0.162 m−2). e results for the shannon index (h′) and pielou evenness index (e) showed very similar values among the sites (table 2). specifically, values of h′ ≥ 3 were computed for sites 1–4 and site 7; in contrast, values of h′ < 3 were obtained for site 5, 6, and 8. equally, the highest values of evenness (0.9 ≤ e ≤ 1) were observed for sites 5, 6, and 8; the lowest (0.8 ≤ e < 0.9) for sites 1–4 and site 7 (see table 2). acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 6 https://www.arpal.liguria.it/ a m b ro sio an d feest / m acro fu n g ian d fo rest h ealth statu s table 2 results of biodiversity indices. site code sr pa pd h′ e d d chao ± sd jack. 1 ± sd bood. ± sd scvi ± sd bi s1 52 606 0.606 3.26 0.82 0.95 18.38 238.2 ± 149.91 78.6 ± 7.58 62.58 ± 3.25 3.13 ± 1.1 45,240.04 s2 45 320 0.32 3.25 0.85 0.94 17.28 58.99 ± 8.67 62.1 ± 5.6 53.13 ± 3.1 2.49 ± 0.72 5,862.38 s3 34 520 0.52 3.01 0.8 0.93 15.63 45.46 ± 8.4 46.35 ± 4.39 39.72 ± 2.34 2.26 ± 0.56 13,663.71 s4 59 162 0.162 3.83 0.81 0.97 37.06 79.9 ± 11.83 79.9 ± 7.35 68.7 ± 3.96 2.44 ± 0.59 2,122.64 s5 35 761 0.761 2.62 0.94 0.87 8.29 38.37 ± 3.21 42.6 ± 3.01 39.27 ± 1.93 2.43 ± 0.6 9,751.66 s6 33 428 0.428 2.88 0.8 0.91 12.11 37.06 ± 4.7 39.6 ± 4.08 36 ± 2.36 2.06 ± 0.55 8,153.99 s7 33 284 0.284 3.15 0.99 0.94 17.44 32.42 ± 0.83 34.85 ± 1.64 34.89 ± 1.79 2.36 ± 0.65 4,247.24 s8 23 203 0.203 2.87 0.9 0.93 14.4 23.85 ± 1.4 25.85 ± 1.64 25.07 ± 1.39 2.43 ± 0.5 2,215.27 sr – species richness; pa – population abundance; pd – population density; h′– shannon index; e – pielou evenness index; d – simpson’s index; d – berger–parker index; chao – nonparametric estimator chao1; jack. 1 – jackknife first order – nonparametric estimator; bood. – bootstrap – nonparametric estimator; scvi – species conservation value index; sd – standard deviation; bi – biomass index. a cta m ycologica / 2021 / vo lu m e 56 / a rticle 567 pu b lish er:po lish b o tan icalso ciety 7 ambrosio and feest / macrofungi and forest health status e results for the simpson’s index (d) and berger–parker index (d) showed the highest value for the beech forest (s4; d = 0.97, d = 37.06), while the lowest was recorded for the evergreen oak woods (s5; d = 0.87, d = 8.29). e results of nonparametric estimators for sr and standard deviation for each site are summarized in table 2. e computed estimators (chao, jackknife first order and bootstrap) showed very close or similar values to the observed species richness (e.g., s8: sr = 23, chao = 23.85 ± 1.40) among the study sites. figure 2 shows the rarefaction curves for all eight sites surveyed. despite differing total sr, all the curves clearly show that a considerable number of species were collected. e flattened final part (the asymptote) of the curve indicates that not many species were missed. e species conservation value index (scvi) value ranged from the minimum in site 6 (scvi = 2.06 ± 0.55) to the maximum value at site 1 (scvi = 3.13 ± 1.1). all sites showed very similar values, except for s1 (the highest value) (table 2). e computed biomass index values (bi) ranged from the lowest (bi = 2,122.64) for site 4 to the highest value obtained for site 1 (bi = 45,240.036) (table 2). cluster analysis (figure 3) was plotted on sr, presence/absence data, and shannon index among all 160 plots. e results clearly show that sites (1–8) formed distinctive clusters, especially based on sr and presence/absence values. moreover, site 2 and site 4 form a single cluster based on both sr and presence/absence data (figure 3a,b). in contrast, the dendrogram based on shannon index showed different clusters among the sites (figure 3c). e results of the correspondence analysis are shown in figure 4, where sites (1–8) were distributed in the multivariate space on the basis of their macrofungal sr, presence/absence of species, and shannon index. e isa results (table 3) emphasize that some macrofungal species have a significant indicator value (iv). specifically, we found that 13 common species were statistically significant: armillaria mellea, boletus aereus, entoloma rhodopolium, gymnopus fusipes, hebeloma crustuliniforme, hydnum rufescens, inocybe geophylla, lactarius zonarius, marasmius oreades, mycena crocata, tricholoma bresadolanum, t. saponaceum, and t. ustale (table 3). table 4 shows the biodiversity indices computed in sites 2, 3, and 4 surveyed over 3 consecutive years (2012, 2013, 2014). appendix s9–appendix s14 list the recorded macrofungal species and the sporocarp abundance in sites 2–4 (2012–2014). for all three sites, we observed the lowest index values during the third survey year (2014) (table 4). appendix s15 summarizes the full climate data of sites 2, 3, and 4. mean monthly temperatures ranged from −2.08 °c (february 2012) to 21.74 °c (august 2012), from 0.48 °c (february 2013) to 21.57 °c (august 2013), and from 3.26 °c (january 2014) to 20.06 °c (august 2014). mean rainfall ranged from 1.8 mm (june 2012) to 348.8 mm (november 2012), from 29.8 mm (june 2013) to 416.8 mm (december 2014), and from 50 mm (september 2014) to 676 (november 2014). e results of pca are shown in table 5. mean temperature and rainfall had a statistically significant influence on macrofungal biodiversity indices (sr, pa, h′, and bi) observed at the three sites (2–4) over 3 consecutive years. finally, in figure 5 and figure 6, climatic data were compared with the selected biodiversity indices, expressed on a logarithmic scale, since they are expressed in different measurements. 4. discussion e results obtained in this study support our first hypothesis, that is, in the same surveyed time – 2013, different mediterranean forests can differ in biodiversity quality index values (table 1). specifically, by comparing species richness results and their estimators (here, chao, jackknife, and bootstrap), we can observe that the silver fir plantation (s1) and the beech forest (s4) show higher sr values than the evergreen and broadleaf oak woods (e.g., s3, s5–s7) or chestnut coppices (s2, s8). however, the estimator results confirmed that despite a large number of species being collected at each site (see rarefaction curves; figure 2), the number of expected species in the acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 8 ambrosio and feest / macrofungi and forest health status figure 2 e rarefaction curves for all the study sites (s1–s8). “sites” in the graphs refer to surveyed plots in each site. (s) – sardinia; (l) – liguria; (t) – tuscany; (c) – campania. sites is consistently underestimated (e.g., for s1). ese results confirm that knowing how many species occur in a community, an ecosystem, or in a geographic area, is still a major task in ecology, even if the sampling approach is suitable (feest, 2006) acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 9 ambrosio and feest / macrofungi and forest health status figure 3 cluster dendrograms of the eight sites based on macrofungal species richness (a), presence/absence (b) of species, and shannon index (c). (s) – sardinia; (l) – liguria; (t) – tuscany; (c) – campania. and sampling effort is exhaustive (ambrosio et al., 2018). sr is traditionally used as a direct indicator of the level of biodiversity, but there is no consensus on how it should be estimated (chiarucci, 2012; xu et al., 2012). to overcome this, ecologists have devised many quantitative indices (e.g., shannon, evenness, simpson’s, and acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 10 ambrosio and feest / macrofungi and forest health status figure 4 graphical representation of correspondence analysis of the eight sites based on macrofungal species richness (on the le), presence/absence (middle) of species, and shannon index (right). table 3 summary of indicator species analysis. species site vegetation forest type iv p armillaria mellea s3 deciduous oak wood natural 1 0.03∗ boletus aereus s7 evergreen oak wood natural 0.655 0.030∗ entoloma rhodopolium s6 evergreen oak wood natural 0.783 0.020∗ gymnopus fusipes s2 chestnut coppice managed 1.000 0.05∗ hebeloma crustuliniforme s4 beech forest natural 0.973 0.01∗∗ hydnum rufescens s8 chestnut coppice managed 0.894 0.005∗∗ inocybe geophylla s2 chestnut coppice managed 0.975 0.05∗ lactarius zonarius s7 evergreen oak wood natural 0.655 0.030∗ marasmius oreades s7 evergreen oak wood natural 0.845 0.005∗∗ mycena crocata s4 beech forest natural 0.964 0.03∗ tricholoma bresadolanum s1 silver fir planted 0.933 0.05∗ tricholoma saponaceum s6 evergreen oak wood natural 0.966 0.005∗∗ tricholoma ustale s8 chestnut coppice managed 0.775 0.035∗ iv – indicator value; p – significance (∗ p < 0.05; ∗∗ p < 0.01). species are ordered alphabetically. berger–parker indices) that combine the effect of the total sr, occurrence, and evenness. our results confirm that the silver fir plantation (s1) and beech forest (s4) showed the highest values of diversity (h′and d), followed by the evergreen and broadleaf oak woods (e.g., s3, s5–s7) and chestnut coppices (s2, s8). data obtained on population abundance (pa in table 1) and density (pd) (here, individuals are fruiting bodies/sporocarps per species) show a different scenario: sites with lower sr and/or h′(e.g., s3) show higher pa or pd values. conversely, sites with high sr and/or h′(e.g., s4) show low pa or pd values, so that sites are species-rich but sporocarps-poor. in addition, biomass index (bi) values do not completely reflect the sr results of sites (only s1 shows the highest sr and bi values). ese results suggest that the ecology and habitat of macrofungi should also be considered. many macrofungi can have a large size, others medium or small size, and they can grow in a dense cluster, gregarious or as “solitaries.” hence, the bi value of a site can provide an indirect indicator of the occurrence of macrofungal communities within the given site. moreover, for the purposes of prioritizing the conservation of sites, based on their level of diversity, compound indices are oen preferred over sr, as the complexity of biodiversity cannot be encapsulated by a single number (ambrosio et al., 2018; feest, 2006; gaston & spicer, 2004). site community similarity was tested by both h-ca and ca considering the value of sr, presence/absence of species, and shannon index in each surveyed plot and site acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 11 ambrosio and feest / macrofungi and forest health status table 4 summary of biodiversity indices in s2, s3 and s4 over three consecutive years (2012–2014). site 2 – chestnut coppice (l) site 3 – deciduous oak wood (l) site 4 – beech forest (l) 2012 2013 2014 2012 2013 2014 2012 2013 2014 sr 42 45 12 29 34 30 36 59 16 pa 394 320 81 188 520 108 354 162 141 pd 0.394 0.32 0.081 0.188 0.52 0.108 0.354 0.162 0.141 h′ 3.16 3.25 2.22 3.14 3.01 2.86 3.19 3.83 2.3 e 0.84 0.85 0.89 0.93 0.8 0.87 0.89 0.81 0.82 d 0.93 0.94 0.87 0.94 0.93 0.92 0.94 0.97 0.87 d 15.5 17.28 8.09 19.31 15.63 12.84 18.5 37.06 7.89 chao ± sd 52.12 ± 6.66 58.99 ± 8.67 14.53 ± 3.33 30.07 ± 1.45 45.46 ± 8.4 33.2 ± 2.93 43.91 ± 6.59 79.9 ± 11.83 24.55 ± 9.68 jack. 1 ± sd 57.2 ± 5.08 62.1 ± 5.6 15.8 ± 1.9 33.75 ± 3.1 46.35 ± 4.39 38.55 ± 2.85 45.5 ± 3.83 79.9 ± 7.35 21.7 ± 3.33 bood. ± sd 49.49 ± 2.9 53.13 ± 3.1 13.93 ± 1.21 28.87 ± 2.94 39.72 ± 2.34 34.9 ± 2.3 40.58 ± 2.16 68.7 ± 3.96 18.49 ± 1.8 scvi ± sd 2.5 ± 0.74 2.49 ± 0.56 2.5 ± 1.5 2.24 ± 0.73 2.26 ± 0.59 2.27 ± 0.73 2.22 ± 0.72 2.44 ± 0.6 2.12 ± 0.5 bi 5,464.77 5,862.38 41,483.95 4,410.71 13,663.71 3,150.59 3,542.11 2,122.64 1,361.97 sr – species richness; pa – population abundance; pd – population density; h′– shannon index; e – pielou evenness index; d – simpson’s index; d – berger–parker index; chao – nonparametric estimator chao1; jack. 1 – jackknife first order – nonparametric estimator; bood. – bootstrap – nonparametric estimator; scvi – species conservation value index; sd – standard deviation; bi – biomass index; l – liguria. table 5 results of principal correspondence analysis. pc1 pc2 r2 p (>r) temperature (°c) 0.81090 0.58518 1 0.000999∗∗∗ rainfall (mm) −0.81090 0.58518 1 0.000999∗∗∗ significance (p): ∗∗∗ ≤0.001. (figure 3 and figure 4). e results show that patterns of site distribution are driven by a geographical gradient (e.g., geographic distance) and vegetation type (figure 3). e dendrograms in figure 3 indicate that sites form distinctive groups on the basis of the vegetation and geographic distance. all these results support our first hypothesis that different mediterranean forest types show different macrofungal biodiversity quality values. both the silver fir plantation (s1) and beech forest (s4) showed different values and patterns of distribution from those of the other sites. conversely, within the same vegetation type, sites show more statistical similarities: the evergreen and broadleaf oak woods (s3, s5–s7) or chestnut coppices (s2, s8) form very close (clustered) groupings. by considering forest management, we observed that unmanaged sites had higher macrofungal biodiversity values than the managed sites (s2, s8). ese results agree with those of other studies (see tomao et al., 2020) that recognize that unmanaged forests (especially when they evolve to old-growth forests) are reserves of high fungal sr and diversity. tomao et al. (2020) also affirmed that some silvicultural practices can negatively affect fungal diversity and dynamics (in our case, s2 and s8). to assess the effect of climate conditions (temperature and rainfall) (our second hypothesis), we compared these factors with various biodiversity indices (sr, pa, h′, and bi) recorded at three sites (2–4) over 3 consecutive years (2012–2014) (figure 5, figure 6, and table 5). both climatic variables were statistically significant in relation to the biodiversity indices (table 5). however, we observed that during the third year of surveys (2014), we measured the lowest index values. overlapping these results with monthly temperatures and rainfall data, we can observe that 2014 was hotter than 2013 and 2012, and the mean rainfall was lower than in the previous years, especially in summer and early fall. ese results confirm that at the same site, surveyed over consecutive time periods, climatic variations can affect the composition of macrofungal communities. several studies (büntgen et al., 2011, 2013; egli, 2011) confirmed that seasonal macrofungal yield is influenced positively by precipitation amounts and mean temperatures, especially by spring–summer acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 12 ambrosio and feest / macrofungi and forest health status figure 5 graphical comparison of mean monthly temperatures detected during each surveyed year (2012–2014) in sites 2, 3, and 4 with macrofungal biodiversity indices (in logarithmic scale). sr – species richness; pa – population abundance; h′ – shannon index; bi – biomass index. temperatures are expressed in °c. temperatures and total precipitation from september to october. inter-annual fluctuations and long-term trends in the formation of fungal fruiting bodies, as well as intra-annual shis in the timing of their productivity, have been related to land/use-cover change (including deforestation), increased harvesting, and pollution (büntgen et al., 2011, 2013; tomao et al., 2020). hence, macrofungi and measurements of their biodiversity can be useful indirect indicators of interand intra-annual climate changes. e specific habitat requirements of fungi and influence of biotic and abiotic factors on their growth make them well suited as indicators for selecting conservation areas and monitoring their status. notoriously, northern european studies (heilmann-clausen et al., 2014) emphasized the role of wood-inhabiting fungi (e.g., polypores) as useful indicators of forest perturbations (e.g., the lack of old trees and dead wood); however, on the basis of our results and other studies performed in south mediterranean forests (i.e., ambrosio & zotti, 2015), we suggest that epigeous agaric, bolete, and gasteromycete macrofungi should be considered as useful habitat indicators (table 3) because they are highly sensitive in detecting changes in ecosystems (e.g., n deposition; feest et al., 2014). moreover, the calculation of a statistical index (scvi; table 2 and table 4) can provide indirect information about the presence of rare species. in our study, site 1 showed a higher scvi value, indicating the presence of rare species, compared to other sites, and its value remained stable (see table 4) over time, despite poor rainfall. several indicator schemes based on fungi have been suggested for assessing the conservation value of forests and grasslands (feest, 2013); however, biodiversity conservation initiatives till date have oen overlooked the pivotal role of fungi in biodiversity monitoring (heilmann-clausen & vesterholt, 2008). acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 13 ambrosio and feest / macrofungi and forest health status figure 6 graphical comparison of mean monthly rainfall detected during each surveyed year (2012–2014) in sites 2, 3, and 4 with macrofungal biodiversity indices (on a logarithmic scale). sr – species richness; pa – population abundance; h′– shannon index; bi – biomass index. rainfall is expressed in mm. in conclusion, the results of this study represent the ecological contribution and role of the quality of macrofungal biodiversity as a predictor of mediterranean forest health and dynamics. e results obtained show that mediterranean forests, while briefly investigated, show different macrofungal biodiversity indices influenced by forest type and management. interand intra-annual temperatures and rainfall fluctuations affected the formation of fungal fruiting bodies (sporocarps) and consequently the dynamics of communities. based on these results and published literature, it is possible to confirm the ecological response to climate variations reported for a broad range of taxa, including macrofungi. preserving fungal diversity in forest ecosystems is important because of the positive relationship between fungi and ecosystem multifunctionality, as well as their potential early role as indicator species. mycorrhizal fungi, in particular, synergistically affect plant communities and ecosystem services through altered functional traits and ecology of host plants (tedersoo et al., 2020). preserving and restoring forests and improving predictions about the impacts of global change and pollution on vegetation and soil processes, by both direct and indirect indicators, means guaranteeing the conservation of the most bio-complex ecosystems and related ecological processes that are the basis of future life on earth. 5. supplementary material e following supplementary material is available for this article: appendix s1: list of recorded species in site 1 (in 2013). appendix s2: list of recorded species in site 2 (in 2013). acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 14 ambrosio and feest / macrofungi and forest health status appendix s3: list of recorded species in site 3 (in 2013). appendix s4: list of recorded species in site 4 (in 2013). appendix s5: list of recorded species in site 5 (in 2013). appendix s6: list of recorded species in site 6 (in 2013). appendix s7: list of recorded species in site 7 (in 2013). appendix s8: list of recorded species in site 8 (in 2013). appendix s9: list of recorded species in site 2 (in 2012). appendix s10: list of recorded species in site 2 (in 2014). appendix s11: list of recorded species in site 3 (in 2012). appendix s12: list of recorded species in site 3 (in 2014). appendix s13: list of recorded species in site 4 (in 2012). appendix s14: list of recorded species in site 4 (in 2014). appendix s15: mean monthly temperatures and rainfall recorded in site 2, 3, and 4 in 2012, 2013, and 2014. acknowledgments authors are grateful to g. nardi for graphic images and to the managing/production editor for linguistic revision. anonymous reviewers are also acknowledged for useful comments and observation. references ambrosio, e., lancellotti, e., brotzu, r., salch, h., franceschini, a., & zotti, m. 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(2012). assessing non-parametric and area-based methods for estimating regional species richness. journal of vegetation science, 23, 1006–1012. https://doi.org/10.1111/j.1654-1103.2012.01423.x acta mycologica / 2021 / volume 56 / article 567 publisher: polish botanical society 17 https://doi.org/10.1016/j.funeco.2014.01.005 https://doi.org/10.1007/s10531-006-9117-7 https://cran.r-project.org/package=vegan https://doi.org/10.2307/2530750 https://doi.org/10.1038/ismej.2014.37 https://doi.org/10.1126/science.1256688 https://doi.org/10.1126/science.aba1223 https://doi.org/10.1038/nature02121 https://doi.org/10.1016/j.foreco.2019.117678 https://doi.org/10.1139/b07-068 https://unstats.un.org/sdgs/indicators/indicators-list/ https://doi.org/10.1111/j.1654-1103.2012.01423.x can macrofungal biodiversity predict forest status and dynamics? a view from south european mediterranean forests (italy) 1 introduction 2 material and methods 2.1 study sites figure 1 2.2 survey procedure table 1 2.3 data sets 2.4 data analyses 2.5 climate data 3 results table 2 4 discussion <no title> figure 2 figure 3 figure 4 table 3 table 4 table 5 figure 5 figure 6 5 supplementary material acknowledgments references the lichen biota of pinus sylvestris under the impact of some stand-related factors: a case study from the south-eastern part of żerków-czeszewo landscape park (wielkopolska-kujawy lowland) acta mycologica article id: 562 doi: 10.5586/am.562 publication history received: 2020-03-19 accepted: 2020-10-02 published: 2021-05-19 handling editor piotr zaniewski; warsaw university of life sciences – sggw, poland; https://orcid.org/0000-00020792-9854 authors’ contributions dzb: field survey, laboratory research, participation in data analysis and interpretation, and manuscript preparation; kk: concept and performing of statistical analysis, participation in interpretation of results, and in manuscript preparation funding the study was financed by the institute for agricultural and forest environment, polish academy of sciences (statutory funds). competing interests no competing interests have been declared. copyright notice © the author(s) 2021. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. original research paper in lichenology the lichen biota of pinus sylvestris under the impact of some stand-related factors: a case study from the south-eastern part of żerków-czeszewo landscape park (wielkopolska-kujawy lowland) daria zarabska-bożejewicz *, krzysztof kujawa institute for agricultural and forest environment, polish academy of sciences, bukowska 19, 60-809 poznań, poland * to whom correspondence should be addressed. email: zardaria@wp.pl abstract a lichenological inventory was conducted in the pine stands in the south-eastern region of żerków-czeszewo landscape park. e aim of this study was to recognize the lichen species diversity on the bark of pinus sylvestris by considering tree age and forest habitat type. in total, 26 species of lichenized fungi were found on the bark of p. sylvestris. e biota comprises common and widespread species in poland. tree age positively and significantly influenced the species richness on the bark of p. sylvestris. e suitability of the pine tree age for lichens evident as alpha diversity seemed to be independent from the habitat of the trees. e age of phorophytes strongly affected the presence of hypocenomyce scalaris. mixed fresh coniferous forests were characterized by higher betaand gamma diversity compared to fresh coniferous forests. ere was no correlation between the number of species per tree (alpha diversity) and diversification of species composition in relation to the distance between pines. e findings indicate that maintenance of a variety of phorophytes and age-diversified tree stands can prevent impoverishment of the lichen biota in the south-eastern part of żerków-czeszewo landscape park. keywords lichenized fungi; species richness; pine; coniferous forest; west-central poland 1. introduction coniferous forests dominate the forest habitat types in poland and occupy 50.1% of the total forest area (zajączkowski et al., 2019). most scots pine stands are managed (kubiak et al., 2015) and human activity considerably affects their physiognomy, species structure, and dynamics (stefańska, 2006). e dominance of scots pine forest has a significant impact on the species diversity in the forest landscape in poland (kubiak et al., 2015). is importance is increased in agricultural landscapes or in areas under other anthropogenic pressures, where the presence of the coniferous forest may allow the survival of many organisms. species diversity of epiphytic lichens in a forest community is determined by a number of factors, including stand-related factors. e effect of some has been recognized. changes in the occurrence and abundance of lichens may appear in response to the impact of the vegetation type (ardelean et al., 2015; giordani et al., 2012; kolanko, 2013; zarabska, 2009), the species and age structure of the tree stands (hauck, 2011; hauck & spribille, 2005; kapusta et al., 2004; kolanko, 2013; kubiak, 2013; kubiak et al., 2016; marmor et al., 2013; sevgi et al., 2019), quality of available acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 1 https://doi.org/10.5586/am.562 https://orcid.org/0000-0002-0792-9854 https://orcid.org/0000-0002-0792-9854 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0003-3881-2904 https://orcid.org/0000-0003-2812-4702 mailto:zardaria@wp.pl zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors substrata (kapusta et al., 2004), and the chemical and physical properties of the bark of phorophytes (hauck, 2011 and literature cited there; hauck & spribille, 2005; kapusta et al., 2004; kolanko, 2013; kubiak, 2013; sevgi et al., 2019). in addition, many studies have proven the influence of microclimatic conditions on the lichenized fungi, including light conditions (giordani et al., 2012; hauck, 2011 and literature cited there; hauck & spribille 2005; kapusta et al., 2004; sevgi et al., 2019), precipitation chemistry, and the input of pollutants (glanc, 1995; hauck, 2011 and literature cited there; hauck & runge, 2002; kapusta et al., 2004), especially so2 and nitrogen pollution (nh3 and nox) (hauck, 2011), topographic variables (ardelean et al., 2015; sevgi et al., 2019), and stand continuity (hauck, 2011 and literature cited there). e lichen biota of pinus sylvestris in relation to its species richness seems to be well recognized based on local or regional inventories of epiphytes. however, factors determining the occurrence of lichens growing on the bark of this phorophyte are less known. e physicochemical properties of the bark of p. sylvestris affect the composition of the lichen biota (e.g., kapusta et al., 2004; zalewska et al., 2004). marmor et al. (2013) described vertical changes in lichens on the bark of pines. some species, including rare taxa, can find favorable conditions for their establishment and persistence in higher parts of the trunk and branches of p. sylvestris (marmor et al., 2013). we decided to focus on human-related factors in forests and their effect on the species richness and composition of lichens on the bark of p. sylvestris. forest management is one of the most important factors affecting the persistence of lichens in forest communities (e.g., kolanko, 2013; kubiak et al., 2016; motiejûnaitë & fałtynowicz, 2005; zaniewski et al., 2014). clear cutting, site preparation, artificial replanting, and regular thinning are all part of the intensive management of scots pine forest in central europe (stefańska-krzaczek, 2012). e effect of forest management on the species richness of the lichen biota is still poorly documented in poland (kubiak, 2013 and literature cited there; kubiak et al., 2016; wilkoń-michalska et al., 1998; zaniewski et al., 2014). to fill this gap in the knowledge we aimed at recognizing the species diversity on the bark of p. sylvestris considering tree age and forest habitat type in the south-eastern part of żerków-czeszewo landscape park. 2. material and methods 2.1. study area żerków-czeszewo landscape park is situated in the central part of the wielkopolska province, where the lutynia and warta rivers join in the warta valley. e park was established in 1994 and its area now measures 15,800 ha within miłosław, nowe miasto nad wartą, and żerków communities (masztalerz, 2014). e park was established to protect the postglacial relief with special attention to the warsaw-berlin ice-marginal streamway (polish: pradolina warszawsko-berlińska) and culmination of the żerków wall (polish: wał żerkowski). ese are valuable ecosystems, that in particular include some oak-hornbeam and alluvial forest associations in the warta valley; rare and protected species of plants, animals and fungi as well as their habitats; and the spatial structure of land considering local landscapes features and valuable culture aspects (resolution of the sejmik of the wielkopolska province, 2013). e study was conducted in the south-eastern part of żerków-czeszewo landscape park, in the forest communities occupying approximately 250 ha between the villages of podlesie, żerniki, and ludwinów-bogiel (figure 1). forests in the study area are dominated by scots pine. e age of these dominant trees did not exceed 98 years. e lutynia river flows along the western edge of the investigated forest communities. a fishpond complex is located in the western part of the study area, and the road from żerków to żerniki runs through the northern part. e agricultural landscape prevails in the surrounding area. in 2015, the mean annual precipitation was 400 mm, and the average annual temperature was approximately 10 °c (institute of meteorology and water management, 2020). acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 2 zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors figure 1 study area: a – forests; b – unforested areas; c – breeding ponds and other water reservoirs; d – the lutynia river and smaller watercourses; e – the border of żerków-czeszewo landscape park; f – roads; g – villages/settlements. żerków-czeszewo landscape park has not been intensively lichenologically explored. data presented in this article were collected during an inventory aimed at recognition of lichens both within the park and its surroundings, and in this part of the wielkopolska-kujawy lowland. e lichenological survey was conducted in the managed forest typical for the polish lowlands considering forest habitat types, the dominant tree, and the type of forest management. 2.2. sampling e study was conducted in 2014 and 2016. sampled trees were selected randomly in the particular forest habitat types (mixed fresh coniferous forest – mfc; humid mixed coniferous forest – hmc; fresh coniferous forest – fc; mixed fresh deciduous forest – mfd; mixed humid deciduous forest – mhd; fresh deciduous forest – fd). eir locations were distributed within the whole study area. source data for forest habitat types and the age of the dominant tree were obtained from an interactive forest map of the local forestry authority (regional directorate of state forests in poznań, 2012). forest habitat type is one of the main typological systems, especially in practical forestry in poland (pielech & malicki, 2014). is classification is based on the comparison of the fertility and the humidity of the soil, climate, and landform features and geological structure in forest areas (bańkowski et al., 2003). is study included mfc, hmc, fc, mfd, mhd, and fd. some select information about their characteristics are given below according to bańkowski et al. (2003). english translations of the original polish name of soil types and subtypes were adopted from kabała et al. (2019). all of the aforementioned forest habitat types are lowland habitat types. mfc includes fresh habitats with mineral soils, under very weak or weak impacts of groundwater. it is mainly found on podzolic rusty soils or typical rusty soils, and much less oen on podzolic soils. hmc includes quite poor habitats, under moderate or quite strong influence of groundwater. it occurs mainly on gley-podzolic soils and gley-podzols. in the wetter variant, hmc can be found on, among others, gley and peat soils. fc is characterized by poor, fresh habitats, under acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 3 zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors very weak or weak influence of waterground. podzolic soils dominate among the fc soils, with arenosols being less frequent. mfd includes moderately fertile, fresh habitats that are very weakly or weakly affected by groundwater or rainwater. typical rusty soils, brown-rusty soils, podzolic clay-illuvial soils, and podzolic brown soils are mainly distinquished. quite fertile and humid habitats, under moderate or quite strong influence of groundwater or rainwater, are typical for mhd. fd includes fertile and fresh habitats, which are very weakly or weakly impacted by groundwater or rainwater. brown soils and clay-illuvial soils are most oen found, while brown-rusty soils, pararendzinas, and black earths are much less common. e proportion of the analyzed forest habitat types reflects their share in the investigated forest communities. altogether, 201 p. sylvestris trees were sampled (mfc: 154; hmc: six; fc: 24; mfd: nine; mhd: six; fd: two). e age of the pines varied from 26 to 90 years. lichens were recorded on the trunk of each tree up to 1.7 m from the ground. when lichen species could not be determined in the field, specimens were collected for further identification in the laboratory using stereoscopic and light microscopy. for the analysis of secondary metabolites in lichen thalli, thin layer chromatography was performed in solvents a and c in accordance with the methods described by culberson and ammann (1979) and orange et al. (2001). e collected specimens are housed in the department of agroecology and bioindication, e institute for agricultural and forest environment (iafe) of the polish academy of sciences in poznań. species nomenclature follows fałtynowicz and kossowska (2016). e nomenclature of cladonia coniocraea is based on diederich et al. (2020). reatened species categories in poland those of cieśliński et al. (2006). 2.3. statistical analyses e difference in tree stand age between forest habitat types was tested using the mann–whitney test. e mantel test was used to check the effect of the distance between surveyed trees, based on the geographic coordinates of surveyed research points, the similarity in the number of lichen species per tree, and the similarity in the lichen species composition on trees. e total lichen species richness was assessed with the use of the indicator chao2-bc (bias corrected). mao tau rarefaction curves were used to quantify relationships between the sample size (number of pines surveyed) and the number of lichen species. a generalized linear model (glz) was used to quantify the relationships between habitat structure and the number of lichen species. in the model, the explained variable was the number of species per tree, and the habitat variables (predictors) were forest habitat type (fht) (factorial variable; fht = mfc or fc), age of the dominant tree (age) (numerical variable), and interaction fht × age. e effect of age and fht (mfc vs. fc) on the presence/absence of most common lichen species (i.e., with more than 10 records) was tested with the use of logistic regression. e spatial variation in the lichen species composition (a measure of beta diversity) was analyzed using the raup–crick distance as a dissimilarity measure (vellend et al., 2007). to describe the patterns of spatial turnover of species, we used the first two principal coordinate axes. e next step of beta diversity analysis was testing the difference in community dispersion between fresh mixed coniferous forests and fresh coniferous forests. datasets of community dispersion (separately for mfc and for fc) consisted of the distances of each sample from a centroid calculated in the principal coordinate space (with first two principal coordinate axes, see above). a permutation f test with 99 permutations was applied to determine if community dispersion was similar in both forest habitat types (anderson, 2006; anderson et al., 2006). e mann–whitney test, glz, and logistic regression were run with the statistica 12.0 (statso). rarefaction mau tao curves were constructed with the use of past 3.22 (hammer et al., 2001). e values of chao2-bc indices were estimated with the use of the r package spader (chao et al., 2016). e assemblage dispersion analysis was performed using r 3.5.2 soware (r core team, 2018) and vegan package (oksanen et al., 2019). acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 4 zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors table 1 lichen species observed, forest habitat type, morphological form, and number of records assigned in the study. species fht morphological form no. amandinea punctata (hoffm.) coppins & scheid. fc, mfd crust. 2 cladonia chlorophaea gr. mfc frut. 1 cladonia coniocraea (flörke) spreng., nom. cons. fc, hmc, mfc, mhd frut. 22 cladonia digitata (l.) hoffm. mfd frut. 2 cladonia fimbriata (l.) fr. mfc frut. 8 cladonia grayi merrill ex sandst. mfc frut. 1 cladonia macilenta hoffm. fc, mfc frut. 3 coenogonium pineti (schrad.) lücking & lumbsch fd, hmc, mfc, mhd crust. 62 evernia prunastri (l.) ach. mfc frut. 2 hypocenomyce scalaris (ach.) choisy fc, hmc, mfc, mfd crust. 137 hypogymnia physodes (l.) nyl. fc, hmc, mfc, mfd, mhd fol. 84 lecania cyrtella (ach.) . fr. mfc crust. 1 lecanora conizaeoides nyl. fc, fd, hmc, mfc, mfd, mhd crust. 167 lepraria elobata tønsberg hmc, mfc crust. 10 lepraria incana (l.) ach. fc, mfc crust. 16 micarea denigrata (fr.) hedl. mfc crust. 7 parmelia sulcata taylor mfc fol. 3 parmeliopsis ambigua (wulfen) nyl. mfc fol. 3 phaeophyscia orbicularis (neck.) moberg mfc, mfd fol. 2 physcia adscendens (fr.) h. olivier fc, fd, mfc, mfd, mhd fol. 9 physcia tenella (scop.) dc. mfc fol. 1 placynthiella icmalea (ach.) coppins & p. james mfc crust. 1 polycauliona polycarpa (hoffm.) frödén, arup & søchting mfc, mfd fol. 2 scoliciosporum chlorococcum (graeve ex stenh.) vězda fc, mfc crust. 2 violella fucata (stirt.) t. sprib. mfc crust. 1 xanthoria parietina (l.) . fr. mfc, mfd fol. 2 fht – forest habitat type; further explanations in: material and methods – sampling. morphological form: crust. – crustose; fol. – foliose; frut. – fruticose. no. – number of records. 3. results 3.1. floristic data overall, 26 lichen species growing on the bark of p. sylvestris were found in the study area. of these species, 11 were crustose (crust.), eight were foliose (fol.), and seven were fruticose (frut.), including cladonia species. e number of species ranged from one to six per tree (mean, 2.7; median, 3). e most frequent were cladonia coniocraea (22), coenogonium pineti (62), hypocenomyce scalaris (137), hypogymnia physodes (84), and lecanora conizaeoides (167). one of the recorded lichens, evernia prunastri, is a near-threatened species in poland (cieśliński et al., 2006). a list of the lichen species found in the investigated area is given in table 1. 3.2. effect of habitat structure on the number of lichen species e study sites were categorized according to the forest habitat types: mfc, hmc, fc, mfd, mhd, and fd. to verify the effect of the age of trees and forest habitat types, an analysis considered the lichens growing on the bark of 178 p. sylvestris trees located in mfc and fc, excluding other forest habitat types because of the small number of samples. e age of tree in mfc forest (31–90 years, median 74 years) was significantly higher compared to fc forest (18–81 years, median 56 years) in the mann–whitney test (z = 3.39, p < 0.001). acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 5 zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors table 2 glz relationships between the number of lichen species per tree, forest habitat type (fht: mfc, fc) and tree stand age (age). effects coefficient 95% ci limits wald’s statistics p value lower upper intercept 0.24 −0.39 0.87 0.57 0.4506 age 0.01 0.00 0.02 5.5 0.0189 fht −0.16 −0.79 0.47 0.24 0.6230 fht × age 0.00 −0.01 0.01 0.11 0.7401 glz – generalized linear model; fht – forest habitat type; mfc – mixed fresh coniferous forest; fc – fresh coniferous forest; ci – confidence interval. figure 2 relationships between the number of lichen species (no. of species, y axis) per tree and the age of tree (age, x axis) in mixed fresh coniferous forests (le) and fresh coniferous forests (right). e spatial distribution of the investigated trees was aggregated. however, the distance between trees did not affect the lichen species composition (mantel statistic r = −0.003, p = 0.53) or the number of lichen species per tree (mantel statistic r = −0.002, p = 0.51). in the glz, the number of lichens was positively affected by tree age (table 2). e effect of pine tree age on lichen alpha diversity (species number per tree) seemed to be independent from the habitat in which tree grew (figure 2), reflected by the insignificant interaction between the forest habitat type and tree age (table 2). among seven common or moderately frequent species (at least 10 records), the age of phorophytes positively affected the presence of h. scalaris, and marginally positively affected the occurrence of c. coniocraea and h. physodes (table 3). e beta diversity of the lichen community differed slightly between mfc and fc forests. in the similarity space, the locations of lichen communities in fc forests were found in the locations of lichen communities in mfc forests (figure 3). e findings indicated that although the lichen community in fc forests consisted of the same species sets as in mfc forests, the dispersion of these species in fc forests was lower (figure 4). e difference was marginally statistically significant in the permutation test for homogeneity of multivariate dispersion (f = 3.11, df = 1, 99 permutations, p = 0.09). e species richness on the bark of pine in mfc forests was more than three times higher (24 species; chao2-bc = 38.9, sd = 12.8) than in fc forests (10 species; chao2-bc = 12.9, sd = 4.0). erefore, despite the lack of statistically significant differences in the number of lichen species per tree in both studied forest habitat types (alpha diversity), the total species richness (the one considered as gamma acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 6 zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors table 3 logistic regression analysis of the relationship between the presence/absence of three common lichen species on pine and forest habitat type (fht: mixed fresh coniferous forest vs. fresh coniferous forest) and pine age (age). effect coefficient 95% ci limits wald test odds ratio 95% ci limits hosmer–lemeshow test lower upper statistics p value lower upper statistics p value cladonia coniocraea intercept. −5.71 −9.30 −2.12 9.71 0.002 fht −0.87 −4.46 2.72 0.22 0.636 0.18 0.00 232.60 age 0.06 0.00 0.11 4.47 0.035 1.06 1.00 1.12 fht × age 0.00 −0.05 0.06 0.02 0.876 10.9 0.09 hypocenomyce scalaris intercept. −4.41 −7.89 −0.92 6.15 0.013 fht −0.75 −4.23 2.74 0.18 0.675 0.23 0.00 238.35 age 0.09 0.03 0.16 8.19 0.004 1.10 1.03 1.17 fht × age 0.00 −0.06 0.06 0.00 0.974 28.3 0 hypogymnia physodes intercept. −4.87 −8.48 −1.25 6.95 0.008 fht 1.53 −2.09 5.15 0.69 0.407 21.40 0.02 29,693.46 age 0.08 0.02 0.14 6.01 0.014 1.08 1.02 1.15 fht × age −0.04 −0.10 0.03 1.23 0.267 9.8 0.13 wald test was used to estimate the individual effect statistical significance. e hosmer–lemeshow test is a measure of the overall model goodness-of-fit assessment. ci – confidence interval. figure 3 ordination diagrams of representing a spectrum of lichen communities recorded in mixed fresh coniferous (mfc) forest (black) and fresh coniferous (fc) forest (red) based on principal coordinate analysis, with the use of presence-absence data and raup–crick similarity distance. diversity) in mfc forests was higher than in fc forests. ese results were also confirmed by rarefaction curves in which the total number of species was higher in mfc forests than in fc forests, independent of the number of investigated trees (figure 5). acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 7 zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors figure 4 boxplots of lichen community dispersion (distance between samples and centroid) estimated with raup–crick distances in mixed fresh coniferous (mfc) forest and fresh coniferous (fc) forest. figure 5 sample-based rarefaction curves (mau tao) for the number of lichen species (thick lines) in fresh coniferous (fc) and mixed fresh coniferous (mfc) forest curves mao tau. in lines show standard deviations of species number. 4. discussion altogether, 26 species of lichenized fungi were recorded on the bark of p. sylvestris in the pine forests in the south-eastern part of żerków-czeszewo landscape park. epiphytic lichen biota in scots pine stands usually comprises few species (e.g., cieśliński, 1997; izydorek, 2010; kolanko & matwiejuk, 2001; kubiak et al., 2016; motiejûnaitë & fałtynowicz, 2005; wolseley et al., 2006). however, the lichen abundance can be considerable (e.g., izydorek, 2010). e physicochemical properties of the bark of p. sylvestris do not favor the development of a rich lichen acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 8 zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors biota (e.g., hauck, 2011 and literature cited there; kapusta et al., 2004; zalewska et al., 2004). e bark of pine is very acidic with poor water holding capacity and is constantly exfoliated (zalewska et al., 2004). e higher number of crustose lichen species we recorded, which constituted 42% of the total number of species, could be attributed to the branch structure of pine. eir presence is promoted by dry substrate conditions on the trunk of pine trees caused by loss of precipitation via the branches compared to the trunk (hauck, 2011 and literature cited there; sevgi et al., 2019 and literature cited there). lichenological surveys on scots pines in estonian coniferous forests revealed a significant correlation between the total lichen species richness on a tree of pine and species richness up to 2 m. such information could be helpful in identifying pine trees that potentially harbor the most species (marmor et al., 2013). most of the lichenized fungi occurring on pines are common and widespread species, which has been confirmed in lichenological inventories in other parts of poland (cieśliński, 1997; fałtynowicz, 1992; faltynowicz & tobolewski, 1989; izydorek, 2010; zalewska et al., 2004). e lichen biota is usually slightly specific, with only a few exclusive species (kubiak et al., 2015). however, according to marmor et al. (2013), many species, including rare lichens, can occupy a higher part of the trunk and canopy of p. sylvestris and therefore can be omitted when lichenological surveys are conducted in the first two metres above the ground. we found a significant positive influence of tree age on the species richness on pines, which was independent of the analyzed forest habitat types. similarly, izydorek (2010) recorded an increase in the number of species and the lichen cover on trunks with the age of conifer trees and decreased canopy closure. kubiak et al. (2016) investigated epiphytic lichen biota within managed forests in northern poland. e results proved that the age of the forest influenced lichen species richness more significantly than the amount of available microhabitat and its heterogeneity. according to svoboda et al. (2010), tree stand age might be a more important factor in areas that are relatively homogenous with regard to other environmental parameters (climate and air pollution). our research was not focused on the recognition of the impact of potential factors affecting lichen species richness. we could not compare their effect on the lichen biota. however, apart from some low local emissions, there were no other important sources of air pollution within, and in the vicinity of, our study area. in addition, climatic conditions seemed to be homogenous. erefore, tree age might be one of the most important factors that significantly determined the species richness of the lichen biota on the bark of p. sylvestris. e physicochemical properties of tree bark change with time (e.g., glanc, 1965). marmor et al. (2013) observed the highest number of lichen species up to approximately 10 m from the ground. according to these authors, the smooth peeling part in the upper part of the trunk discourages the presence of lichens (marmor et al., 2013). is pine bark structure prevails in younger pine stands, and a lower number of species might be associated with this factor. additionally, changes in microclimatic conditions in the tree stands can also occur with age. young pine plantations are more shaded, while older tree stands provide more stabilized microclimatic conditions. in the latter, decreasing tree density results in increased canopy openness and improved light conditions on the lower part of the trunk. is favors the development of epiphytic lichen biota [cf. kubiak et al. (2016) and literature cited there]. however, the dependence of the species richness increase on forest age does not seem so straightforward. in the course of species succession observed on the bark, certain lichen species die out and some new species may appear. a study conducted in a pine forest revealed that the number of species increased in forest stands classified according to their age up to 80 years and then subsequently decreased (fałtynowicz, 1992; fałtynowicz, personal communication, october 8, 2020). in our study area, h. scalaris was one of the most frequently recorded species. e presence of this species was strongly influenced by the age of phorophytes. h. scalaris prefers the dry bark of pinus (diederich et al., 2020). is species was found to be an indicator for old forests during lichenological inventories in managed mediterranean black pine forests (sevgi et al., 2019). since h. scalaris seems to avoid shaded habitats (nimis & martellos, 2017), increased sunlight due to canopy openness and decreasing tree density in older coniferous forests (cf. kubiak et al., 2016) could have promoted the presence of this lichen in our study area. acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 9 zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors e forest habitat type usually reflected by the dominant tree species may affect the diversity of lichens growing on the bark of phorophytes. although we did not detect differences in the mean number of species per tree (alpha diversity) when comparing forest habitat types, mfc forests were characterized by higher betaand gamma diversity compared to fc forests. giordani et al. (2018) conducted biomonitoring surveys. e authors recommended using beta diversity and similarity to analyze temporal and spatial variation in lichen diversity to evaluate anthropogenic impacts. of note, our analyses were restricted to very similar forest habitat types. lack of differences in alpha diversity can also be associated with the sole consideration of lichens growing on pines. tree species influence changes in the occurrence and abundance of lichenized fungi (hauck & spribille, 2005; kapusta et al., 2004; kolanko, 2013; kubiak, 2013; kubiak et al., 2016; marmor et al., 2013). e increase in diversity is related to the different site conditions accompanied by diversified tree species composition in a forest stand (hauck, 2011 and literature cited there). higher variation in species composition (beta diversity) and higher species richness (gamma diversity) in mfc forests can be associated with a higher microhabitat variability that is caused by the presence of more tree species. is favors the establishment and maintenance of a more diverse lichen biota in mfc forests, including those growing on pines. an increase in the species number could be supported by the occurrence of deciduous trees in coniferous communities (cf. izydorek, 2010). diversification of phorophytes, especially the presence of deciduous trees (e.g., oaks, birches, and beeches), in coniferous forests can contribute to the enrichment of the lichen biota (cieśliński, 1997; izydorek, 2010; kubiak et al., 2015; kubiak et al., 2016 and literature cited there; sevgi et al., 2019). presently, spore lichens from sources other than pine phorophytes might also have inhabited the bark of pines in the investigated forest sites. differences in betaand gamma diveristy could also be partially related to the age of the tree, which was significantly higher in mfc forests compared to fresh coniferous forests. we proved that the age of trees positively and significantly influenced the species richness on the bark of p. sylvestris. since the level of air pollution in the study area does not seem to be detrimental to the species richness of lichens (cf. svoboda et al., 2010), we expect that human-related forest activities mainly affected the presence of this group of organisms. e anthropogenic impact on the lichen biota in the forest related to forest management can reduce the occurrence, abundance, and vitality of lichens. in the forest stands we studied, p. sylvestris hosted a moderate number of lichenized fungus species. e negative effect of monoculture on the diversity and persistence of the lichen biota, manifested by the disappearance or decrease in abundance of numerous species, has been documented by many authors (e.g., dingová košuthová et al., 2013; fałtynowicz & tobolewski, 1989; motiejûnaitë & fałtynowicz, 2005). anthropogenic activity in forest ecosystems (i.e., forest management) can affect lichens directly through the destruction of substrates together with lichens growing on them (relevant mainly for epiphytic lichens, less so for terricolous lichens) and/or reduction of the available substrates, such as wood, old trees, and changes in habitat conditions. old trees provide suitable microhabitats for the occurrence of many epiphytic lichens, including rare species (e.g., hauck, 2011 and literature cited there, kubiak, 2013; kubiak et al., 2016). as discussed earlier, both species composition of tree stands and changes in the bark properties with the age of phorophytes can be factors that determine the persistence of the lichen biota and its species richness. age-diversified tree stands enriched the lichen biota growing on pine in the study area, since different species of lichenized fungi can occur in various stages of forest development and some species can be replaced by others. protection of an area, which strengthens the species protection of lichens, allows the preservation of suitable habitat conditions for the development of lichen biota (cf. fałtynowicz, 2006). e study area is located within żerków-czeszewo landscape park. effective protection of lichenized fungi and the maintenance of suitable habitats to prevent impoverishment of the lichen biota in the south-eastern part of żerków-czeszewo landscape park should be primarily based on the careful forest management acta mycologica / 2021 / volume 56 / article 562 publisher: polish botanical society 10 zarabska-bożejewicz and kujawa / lichen biota of pinus sylvestris influenced by stand-related factors activities, such as the maintenance of a variety of phorophytes and age-diversified tree stands. references anderson, m. j. 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https://doi.org/10.1111/j.1365-2745.2007.01233.x https://doi.org/10.1017/s0024282906006190 https://doi.org/10.12775/eq.2014.015 https://doi.org/10.2478/v10111-009-0041-y the lichen biota of pinus sylvestris under the impact of some stand-related factors: a case study from the south-eastern part of żerków-czeszewo landscape park (wielkopolska-kujawy lowland) 1 introduction 2 material and methods 2.1 study area figure 1 2.2 sampling 2.3 statistical analyses table 1 3 results 3.1 floristic data 3.2 effect of habitat structure on the number of lichen species table 2 figure 2 table 3 figure 3 figure 4 figure 5 4 discussion references new data to the knowledge of macrofungi of wolin national park 1 of 22published by polish botanical society acta mycologica original research paper new data to the knowledge of macrofungi of wolin national park małgorzata stasińska*, zofia sotek department of botany and nature conservation, center for molecular biology and biotechnology, environmental testing laboratory, university of szczecin, felczaka 3c, 71-412 szczecin, poland * corresponding author. email: stasinsk@univ.szczecin.pl abstract this paper presents the results of mycological studies conducted in the wolin national park from july to november 2012, and sporadically in the following 4 years. explorations were made by a route method over the whole area of the park, mainly in forest associations: cephalanthero rubrae-fagetum, galio odorati-fagetum, luzulo pilosae-fagetum, and fago-quercetum petraeae. in total, 322 taxa of macrofungi were found, 37 ascomycota and 285 basidiomycota. two of them, hericium coralloides and inonotus obliquus, are under partial protection, 39 on the red list of fungi in poland. for the first time, russula torulosa, previously not reported from poland, was found in wolin national park. among the examined phytocoenoses, galio odorati-fagetum and luzulo pilosae-fagetum are characterized by the highest species variety and abundance. keywords macromycetes; rare species; protected and threatened fungi; protected area; poland introduction national parks belong to strategic natural resources of poland [1]. they are established on the areas of exceptional natural value in order to preserve biological diversity of all groups of organisms, including rare and threatened with extinction species. richness of species in particular national parks has not been sufficiently defined and in the case of fungus biota hardly recognized. until 2000, detailed mycological observations have been conducted only in a few of them (e.g., [2–4]). the majority of the objects under this form of protection have been studied in recent years (e.g., [5–12]). however, the state of recognition of diversity fungi in many polish national parks is still not enough. the wolin national park (wnp) was established in 1960 [13] on the area of ca. 4691 ha, to protect one of the most valuable natural areas of polish coast. it included high coastal cliffs up to 90 m above sea level, moraine hills overgrown with forest complexes, a beach belt and inland lakes (czajcze, domysłowskie, rabiąż, warnowskie, grodno lakes and an artificial reservoir – turkusowe lake). in 1996, the park was extended by inclusion of: 1 nautical mile wide belt of coastal waters of pomeranian bay, islands in the świna reverse delta together with surrounding waters of szczecin lagoon and the lakes, wicko duże and wicko małe [14]. thus, the park has become the first national park of maritime nature protecting the waters of the baltic sea. currently, it covers 10 937 ha, where forest ecosystems occupy 4648.53 ha (42.50% of the area), water ecosystems 4681.41 ha (42.80%), and non-forest ecosystems 1607.46 ha (14.70%). total area of 498.72 ha (4.56%) is under strict protection. there is a great doi: 10.5586/am.1089 publication history received: 2016-12-15 accepted: 2016-12-23 published: 2017-01-23 handling editor tomasz leski, institute of dendrology, polish academy of sciences, poland authors’ contributions both authors participated in the field research and contributed to manuscript preparation. funding the study was financed by the university of szczecin as part of individual research grants. competing interests no competing interests have been declared. copyright notice © the author(s) 2017. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation stasińska m, sotek z. new data to the knowledge of macrofungi of wolin national park. acta mycol. 2016;51(2):1089. http:// dx.doi.org/10.5586/am.1089 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:stasinsk%40univ.szczecin.pl?subject=new%20data%20to%20the%20knowledge%20of%20macrofungi%20of%20wolin%20national%20park http://dx.doi.org/10.5586/am.1089 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1089 http://dx.doi.org/10.5586/am.1089 2 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park variety of landscape forms in the park: the baltic coast, the wolin terminal moraine, lubińsko-wapnickie hills, the wolin lakeland, the dargobądzka plain, the shore of szczecin lagoon, and the świna reverse delta [15]. the predominant elements of the relief are moraine hills covering approximately 75% of inland area of the park. the height difference within the park is from 0 to 116 m (the grzywacz hill). one of the decisive natural values for establishing wnp were forests, first of all, best preserved beech forests occurring here in three kinds of beech wood: acidophilous beech (luzulo pilosae-fagetum), fertile beech wood (galio odorati-fagetum), and orchid beech wood (cephalanthero rubrae-fagetum). beech forest covers the highest points of moraine hills. acidophilous beech occupies the largest area, whereas more species-rich fertile beech occurs on more fertile soils. the rarest orchid beech forest can be seen as a narrow belt on the top of the cliff on a unique soil, cliff naspa. the second important forest group of this area, i.e., the southern and central part of the park, consists of mixed forests most frequently represented by fago-quercetum petraeae, scarcely by betulo pendulae-quercetum roboris, covering small areas on soils with a relatively high water level. another important forest group of wnp comprises empetro nigri-pinetum, linked to sandy soils of costal dunes and leucobryo-pinetum growing on sandy, slightly more fertile, poor in moisture, soils. the other types of forests such as alder, riparian, and vaccinio uliginosi-betuletum pubescentis forests [16], sporadically appear on small areas of the park. due to exceptional natural values, the wolin national park was included into the european network natura 2000 and the habitat area plh320019 of wolin and uznam as well as the bird protection area plb320002 delta świny. the fungus biota of wnp has not been sufficiently recognized. there are only a few studies on macrofungi of that area. first reports of several species from the area of the present park come from german publications which appeared before world war ii [17,18]. at the beginning of 1950’s, czubiński and urbański [19] in their description of diversified flora and fauna mention also the fungi of the planned wolin park. analyzing of mycotrophism in beech communities, dominik [20] took into consideration the chalk promontory near lubin and wapnica, within the borders of wnp. a more detailed study of macrofungi of the park [2] appeared almost 30 years after the first information on the occurrence of macromycetes. however, there were several reports of the presence of single stands of some fungal species from the area of wnp [21–27]. the aim of this paper is to present the new data supplementing the present knowledge of taxonomic diversity and abundance of macrofungi within wnp. methods mycological observations in the wolin national park were made from july to november 2012, may and september 2013 and 2014, and also september of 2015 and 2016. the studies were conducted by the route method, over the area of the whole park, mainly in the forest associations: cephalanthero rubrae-fagetum, galio odorati-fagetum, luzulo pilosae-fagetum, and fago-quercetum petraeae, in five areas under strict protection: (i) prof. wł. szafer, (i) dr b. dyakowski, (iii) prof. z. czubiński, (vi) prof. m. raciborski, (v) doc. s. jarosz (fig. 1). the studies were also carried out in the vicinity of: grodno, międzyzdroje, wapnica, wicko, wisełka, and zalesie, in the region of gosań mt, kawcza mt, enclousure of european bisons, and czajcze lake. the specimens were identified by examining their macroscopic and microscopic features using standard methods for studying macrofungi and references by breitenbach and kräzlin [28], romagnesi [29], kränzlin [30], bernicchia and gorjón [31], and knudsen and vesterholt [32]. the fungal nomenclature and its synonyms are given according to mycobank [33] and index fungorum [34]. the names of vascular plants in the present paper follows mirek et al. [35], and of plant communities, matuszkiewicz [36]. the collected material is deposited in the herbarium of the department of botany and nature conservation, szczecin university (szub-f), poland. 3 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park results during the studies on the area of the wolin national park, 322 taxa of macrofungi were identified, among them: 37 ascomycota and 285 basidiomycota. two of recorded taxa: hericium coralloides and inonotus obliquus, are under partial protection [37] and 39 on the red list of fungi in poland [38]. the category extinct (e) is represented by three species: geastrum rufescens, gloeoporus dichrous, and rhodonia placenta. five taxa: amanita virosa, coprinus picaceus, hericium coralloides, ischnoderma resinosum, and xylobolus frustulatus belong to the category vulnerable (v). the largest group consists of the species classified as rare (r) – 27 taxa, e.g., hydropus subalpinus, mycena crocata, and porodaedalea pini, and only four of indefinite threat (i): hebeloma radicosum, hygrophorus hypothejus, mycena pelianthina, and leratiomyces squamosus. one of the taxa, i.e., russula torulosa, whose fruiting bodies occurred on a grey dune [39], has not been recorded up till now. the greatest richness and diversity of species among the examined phytocoenoses were found in galio odorati-fagetum (148 species) and luzulo pilosae-fagetum (232 species). the following abbreviations are used in the list of species: sp – strict protection area: wssp – prof. wł. szafer; bdsp – dr b. dyakowski; zcsp – prof. z. czubiński; mrsp – prof. m. raciborski; sjsp – doc. s. jarosz. species reported earlier by: ml – lisiewska [2]; ms – stier [18]; zcju – czubiński and urbański [19]; td – dominik [20]; as – skirgiełło [21]; mł – ławrynowicz [23]; ar – ronikier [24]; ww – wojewo da [25]; sf – friedrich [27]; and mst – stasińska et al. [39]. plant community: cf – cephalanthero rubrae-fagetum; fq – fago-quercetum petraeae; gf – galio odorati-fagetum; lf – luzulo pilosae-fagetum. category of threat (according to wojewoda and ławrynowicz [38]): e – endangered; v – vulnerable; r – rare; i – indeterminate. * – taxon new for poland; p – partially protected species; div – forest division; may, aug 2012 – observation date. fig. 1 localization of observation sites in the wolin national park. protected areas: 1 – prof. wł. szafer; 2 – dr b. dyakow ski; 3 – prof. z. czubiński; 4 – prof. m. raciborski; 5 – doc. s. jarosz. 4 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park list of species – ascomycota annulohypoxylon multiforme (fr.) y. m. ju, j. d. rogers & h. m. hsieh [= hypoxylon multiforme (fr.) fr.] – on branches of betula; lf; div 15; aug 2012. ascocoryne cylichnium (tul.) korf – on logs and stumps of fagus; lf; div 2 (sjsp); 25 (mrsp); 48; nov 2012. a. sarcoides (jacq.) j. w. groves & d. e. wilson – on wood; gf; div 125 (bdsp); oct 2012; ml. bertia moriformis (tode) de not. – on twigs of fagus; cf, lf, gf; div 11, 12 (zcsp); 13 (gosań mt); 15b; 16; 25 (mrsp); 30; 46; 67; 68; 89; 104; 114; 109; 128ay; 125 (bdsp); jul–nov 2012; may 2013–2014; sep 2016. bisporella citrina (batsch) korf & s. e. carp. – on branches of fagus; fq, lf, gf; div 2 (sjsp); 15; 15b; 16; 25 (mrsp); 30; 40; 48; 68; 69; 89; 91; 104; 110; 111; 116; 121; 125 (bdsp); 127 (wssp); aug, oct–nov 2012; sep 2014. bulgaria inquinans (pers.) fr. – on logs of quercus; gf; div 88; 105; 125 (bdsp); aug, oct 2012. colpoma quercinum (pers.) wallr. – on twigs of quercus; lf; div 13 (gosań mt); 48; 68; nov 2012; may 2013; sep 2016. dasyscyphus willkommii (hartig) rehm (1881) [= lachnellula willkommii (hartig.) dennis] – on twigs of larix; lf; div 114; sep 2012. dasyscyphus virgineus (batsch) gray [= lachnum virgineum (batsch) p. karst.] – on litter; lf; div 13, 14 (gosań mt); zielonka hill; may 2013–2014; ml. dialonectria episphaeria (tode) cooke [= nectria epishaeria (tode) fr.] – on basidiocarps of diatrype stigma; lf; div 46; 114; sep 2012, 2016. diatrype disciformis (hoffm.) fr. – on branches of fagus; fq, cf, lf, gf; div 2 (sjsp); 11 (zcsp); 13, 14 (gosań mt); 15; 15b; 16; 46; 48; 67; 68; 88; 89; 94; 97; 113; 121; 127 (wssp); jul–nov 2012; may 2013–2014; sep 2016. d. stigma (hoffm.) fr. – on twigs of fagus and quercus; fq, cf, lf gf; div 1c; 11, 12 (zcsp); 15; 25 (mrsp); 30; 46; 48; 68; 104; 113; 114; 125 (bdsp); jul–nov 2012; sep 2014, 2016. diatrypella quercina (pers.) cooke – on branches of quercus; lf; div 25 (mrsp); 46; 48; 68; oct–nov 2012; sep 2016. d. verrucaeformis (ehrh.) nitschke – on branches of betula; lf; div 1b; 1c; 2g,n; 62; 68; 91; 111; aug–nov 2012; sep 2016. elaphocordyceps ophioglossoides (ehrh.) g. h. sung, j. m. sung & spatafora [=  cordyceps ophioglossoides (ehrh.) link] – on basidiocarps of elaphomyces; div 115c; oct 2012; r; ml. elaphomyces asperulus vittad. – below ground; lf; div 2 (sjsp); 14o (gosań mt); aug, nov 2012; mł. e. muricatus fr.– below ground; lf; div 47f; nov 2012; mł. eutypa spinosa (pers.) tul. & c. tul. – on logs of fagus; lf; div 14 (gosań mt); 109a; may 2013–2014. 5 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park gyromitra esculenta (pers.) fr. – on ground; zielonka hill; may 2013. helvella crispa (scop.) fr. – on ground; lf; div 15bj (kawcza mt); 20; 70; 71; aug, oct–nov 2012; ml. h. lacunosa afzel. – on ground; lf; div 2j (sjsp); 89a; oct–nov 2012; r; ml. humaria hemisphaerica (hoffm.) fuckel – on ground; lf, gf; div 14 (gosań mt); 128a, 128az (wssp); aug 2012; ml. hymenoscyphus calyculus (fr.) w. phillips – on wood; lf, gf; div 68; 121; oct 2012. h. fructigenus (bull.) gray – on acorns of quercus; div 14 (gosań mt); 69; may 2013; sep 2014. hypoxylon fragiforme (pers.) j. kickx f. – on branches and logs of fagus; fq, cf, lf, gf; div 2 (sjsp); 9; 11 (zcsp); 13 (gosań mt); 15; 15b; 16; 25 (mrsp); 31; 46 48; 67; 68; 88; 89; 91; 111; 113; 116; 121; 127 (wssp); 125 (bdsp); 128ay; jul–nov 2012; may, sep 2014; sep 2016. kretzschmaria deusta (hoffm.) p. m. d. martin [= ustulina deusta (hoffm.) maire] – on stumps of fagus; lf, gf; div 13; 14 (gosań mt); 15; 25 (mrsp); 67; 68; 89; 121; 128a, 128az (wssp); jul–nov 2012; may 2013; sep 2016; ml. mollisia cinerea (batsch) p. karst. – on wood; lf, gf; div 15; 15a; 15b; 16; 40; 127, 128a, 128az (wssp); aug 2012. nectria cinnabarina (tode) fr. – on twigs; fq, lf, gf; div 15b; 16; 69; 125 (bdsp); aug, oct 2012; sep 2014. otidea alutacea (pers.) massee – on ground; div 70; sep 2014; ml. peziza badia pers. – on ground; div 98; aug 2012; ml. p. micropus pers. – on logs of fagus; lf, gf; div 89; 91; 111; 125 (bdsp); 128a, 128az (wssp); aug, oct 2012. p. vesiculosa bull. – on ground; lf; div 14 (gosań mt); 15b; 16; aug 2012. rutstroemia firma (pers.) p. karst. – on twigs of quercus; gf; div 125 (bdsp); oct 2012. scutellinia scutellata (l.) lambotte – on twigs; div 70m (n lakeside of czajcze lake); sep 2014. xylaria carpophila (pers.) fr. – on cupules of fagus; lf, gf; div 2 (sjsp); 13; 14 (gosań mt); 15b; 16; 46; 68; 94; 114; 127 (wssp); aug–nov 2012; may 2013; sep 2016. x. hypoxylon (l.) grev. – on wood; lf, gf; div 25 (mrsp); 31; 40; 48; 68; 125 (bdsp); jul–nov 2012; sep 2016; ml. x. longipes nitschke – on wood; lf; div 94; 114; sep 2012. basidiomycota agaricus silvicola (vittad.) peck – on ground; lf, gf; div 114; 128ay; aug, sep 2012; ml. 6 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park a. squarrosus oeder [= pholiota squarrosa (vahl) p. kumm.] – at the base of fagus; gf; div 125 (bdsp); oct 2012. amanita citrina (schaeff.) pers. – on ground; lf, gf; div 2 (sjsp); 11, 12 (zcsp); 30; 31; 32; 46; 68; 89; 91; 93; 94; 97; 98; 110; 111; 113; 114; 115; 121; 125 (bdsp); sep–nov 2012; sep 2016; ml, as. a. fulva pers. – on ground; fq, lf, gf; div 1b; 1c; 8; 9; 13 (gosań mt); 15; 16; 17; 39; 40; 62; 70; 89; 91; 93; 94; 97; 98; 104; 105; 110; 111; 113; 114; jul–nov 2012; sep 2014; ml. a. gemmata (fr.) bertill. – on ground; lf; div 1a; 1b; 15a; 40; aug 2012; ml. a. muscaria (l.) lam. – on ground; div 70 (n lakeside of czajcze lake); sep 2014; ml. a. pantherina (dc.) krombh.– on ground; lf; div 15; 39; aug 2012; ml. a. phalloides (fr.) link – on ground; lf; div 31; 88; 94; 104; 105; 114; 115; aug, sep 2012; ml, as. a. porphyria alb. & schwein. – on ground; div 91; 111; oct 2012; ml. a. rubescens pers. – na ziemi; lf, gf; div 8; 13, 14 (gosań mt); 15b; 16; 40; 62; 93; 104; 105; 115; 127 (wssp); jul–sep 2012; ml. a. virosa bertill. – on ground; div 91; 111; oct 2012; v. armillaria solidipes peck [= a. ostoyae (romagn.) herink] – on wood; lf, gf; div 2 (sjsp); 11, 12 (zcsp); 25 (mrsp); 48; 89; 109; 125 (bdsp); oct–nov 2012. auricularia auricula-judae (bull.) j. schröt. – on trunks of fagus and quercus; lf, gf; div 1c; 13 (gosań mt); 16; 127 (wssp); 128ag; aug, oct 2012. auriscalpium vulgare gray – on cone of pinus; fq, lf, gf; div 1a; 1b; 1c; 2g, n; 8; 9; 11, 12 (zcsp); 14 (gosań mt); 25 (mrsp); 30; 31; 32; 39; 41; 69; 91; 104; 111; 121; 125 (bdsp); jul–nov 2012; sep 2014; ml. baeospora myosura (fr.) singer – on cone of pinus; lf; div 8; 14 (gosań mt); 25g (mrsp); 115; aug, oct–nov 2012. bjerkandera adusta (willd.) p. karst. – on wood of fagus and quercus; lf, gf; div 15; 15b; 16; 46, 68; 110; 116; 125 (bdsp); 127, 128az (wssp); aug–nov 2012; sep 2016. bolbitius reticulatus (pers.) ricken – on logs of fagus; lf; div 91f; oct 2012; r. boletus calopus pers. – on ground; lf; div 16w; aug 2012; r. b. edulis bull. – on ground; lf, gf; div 15bj (kawcza mt); 68; 127 (wssp); aug, oct 2012; ml, ms. b. erythropus pers. – on ground; gf; div 125 (bdsp); oct 2012; ml. b. pulverulentus opat. – on ground; div 15ac; aug 2012; r. calocera cornea (batsch) fr. – on wood of deciduous trees; lf, gf; div 2 (sjsp); 17; 48; 68; 69; 104; 110; 127, 128a, 128az (wssp); aug, oct–nov 2012; sep 2016. 7 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park c. viscosa (pers.) fr.– on stumps of pinus; fq, lf, gf; div 1b; 1c; 2 (sjsp); 8; 9; 14 (gosań mt); 31; 39; 40; 41; 62; 104; 111; 115; 128ah; aug–nov 2012; sep 2014; ml. calvatia excipuliformis (scop.) perdeck – on ground; gf; div 128ah; aug 2012. cantharellus cibarius fr. – on ground; lf; div 11, 12 (zcsp); 15b; 16; 40; 41; 88; 91; 104; 111; aug, oct 2012; ml. c. cinereus pers. – on ground; lf, gf; div 104; 127 (wssp); aug 2012. c. tubaeformis fr. – on ground; lf; div 91; 111; oct 2012; ml. chlorophyllum rhacodes (vittad.) vellinga [= macrolepiota rhacodes (vittad.) singer var. rhacodes] – on ground; lf, gf; div 68; 125 (bdsp); oct 2012. chondrostereum purpureum (pers.) pouzar – on log of fagus; div 30; aug 2012. chroogomphus rutilus (schaeff.) o. k. mill. – on ground; lf; div 1a; 1b; 91; 111; 113; 114; aug–oct 2012; ml. clavulina cinerea (bull.) j. schröt. – on ground; lf; oddz. 109; oct 2012. c. coralloides (l.) j. schröt. – on ground; gf; div 121; 127 (wssp); aug 2012; ml. clitocybe clavipes (pers.) p. kumm. – on ground; lf; div 2 (sjsp); 91; 111; 70; oct– nov 2012; sep 2014; ml. c. dealbata (sowerby) p. kumm. – on litter; gf; div 125 (bdsp); oct 2012. c. geotropa (bull. ex dc.) quél.– on ground and litter; lf; oddz. 15b; 16; aug 2012; ml. c. metachroa (fr.) p. kumm. – on litter; lf, gf; div 48; 125 (bdsp); oct–nov 2012. c. nebularis (batsch) p. kumm. – on ground; lf, gf; div 2 (sjsp); 25 (mrsp); 48; 125 (bdsp); oct–nov 2012; ml. collybia cirrata (schumach.) quél. – on debris of fungi; gf; div 125 (bdsp); oct 2012. coltricia perennis (l.) murrill – on ground; fq, lf; div 70; 105; 114; aug, sep 2012; sep 2014; ml. coniophora arida (fr.) p. karst. – on trunk of pinus; div 115; oct 2012. conocybe subovalis kühner & watling – on ground; lf; div 91; oct 2012. coprinellus domesticus (bolton) vilgalys, hopple & jacq. johnson [= coprinus domesticus (bolton) gray] – on stumps; lf, gf; div 13, 14 (gosań mt); 109; 127 (wssp); aug, oct 2012. c. impatiens (fr.) j. e. lange [= c. impatiens (fr.) quél.] – on liter; lf; div 2 (sjsp); nov 2012. c. micaceus (bull.) vilgalys, hopple & jacq. johnson [= c. micaceus (bull.) fr.] – on stumps and logs; lf, gf; div 2 (sjsp); 15b; 16; 125 (bdsp); aug, oct–nov 2012. 8 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park c. xanthothrix (romagn.) vilgalys, hopple & jacq. johnson (= c. xanthothrix romagn.) – on litter; lf; div 2 (sjsp); nov 2012. coprinopsis picacea (bull.) redhead, vilgalys & moncalvo [= c. picaceus (bull.) gray] – on ground; trzciągowo (near div 131g); oct 2012; v. coprinus comatus (o. f. müll.) pers. – on ground; div 70; 98; 114; 128b; sep 2012, 2014; ml. cortinarius bolaris (pers.) fr. – on ground; lf, gf; div 104; 105; 121; aug, oct 2012; ml. c. caperatus (pers.) fr. [= rozites caperatus (pers.) p. karst.] – on ground; lf; div 91; 115; oct 2012; ml, as. c. citrinus j. e. lange ex p. d. orton – on ground, under fagus and quercus; lf; div 116b; oct 2012. c. flexipes (pers.) fr. – on ground; lf; div 2 (sjsp); 11, 12 (zcsp); 31; 91; 110; oct– nov 2012; ml. c. orellanus fr. – on ground; lf; div 15; aug 2012; r. c. paleaceus (weinm.) fr. – on ground; lf; div 115; oct 2012. c. semisanguineus (fr.) gillet – on ground; div 64; sep 2014; ml. c. stillatitius fr. – on ground; lf; div 11h (zcsp); oct 2012. c. torvus (fr.) fr. – on ground; lf; div 91; oct 2012. cratherellus cornucopioides (l.) pers. – on ground; lf; div 14 (gosań mt); aug 2012; ml. crepidotus variabilis (pers.) p. kumm. – on litter; lf, gf; div 1c; 2 (sjsp); 14, 13 (gosań mt); 15; 25 (mrsp); 30; 31; 32; 89; 91; 104; 110; 111; 125 (bdsp); 127 (wssp); aug–nov 2012. crucibulum laeve (huds.) kambly – on twig; div 20; nov 2012; ml. cyathus striatus (huds.) willd. – on litter; lf, gf; div 13, 14 (gosań mt); 15b; 16; 128a, 128az (wssp); aug 2012; ml. cystoderma amianthinum (scop.) fayod – on ground; lf; div 2g; 15; 20; 30; 32; 97; aug–nov 2012; ml. c. carcharias (pers.) fayod – on ground; div 2g; 20; nov 2012; ml. dacrymyces stillatus nees – on wood; fq, lf, gf; div 1a; 1b; 1c; 2g,n; 8; 9; 13 (gosań mt); 15; 16; 30; 31; 39; 40; 41; 48; 62; 70; 89; 91; 104; 111; 114; 127, 128a, 128az (wssp); 128ah; jul–nov 2012; sep 2014. daedalea quercina (l.) pers. – on stump of quercus; lf; div 105; aug 2012. daedaleopsis confragosa (bolton) j. schröt. – on wood of fagus; fq, lf; div 39; 41; 62; 70; 93; 110; 113; aug–nov 2012; sep 2014. datronia mollis (sommerf.) donk – on logs of fagus; lf, gf; div 15a; 16; 25 (mrsp); 113; 125 (bdsp); aug–nov 2012. 9 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park dentipellis fragilis (pers.) donk – on log of fagus; gf; div 109b; oct 2012; (leg. m. walczak); r. entoloma cetratum (fr.) m. m. moser – on ground; div 64; 65; sep 2014; ml. e. juncinum (kühner & romagn.) noordel. – on ground; lf, gf; div 16; 104; 127 (wssp); aug 2012; r; ml. exidia truncta fr. [= e. glandulosa (bull.) fr.] – on wood of quercus; lf; div 2 (sjsp); 25 (mrsp); 41; 48; aug, nov 2012; r. e. plana donk – on wood of deciduous trees; cf, lf; div 11 (zcsp); 13 (gosań mt); 15; 25 (mrsp); 113; 114; aug–nov 2012; may 2013–2014. fomes fomentarius (l.) fr. – on logs and trunks of fagus and quercus; fq, lf, gf; div 1b; 1c; 2 (sjsp); 13, 14 (gosań mt); 15; 15b; 16; 25 (mrsp); 30; 46; 48; 68; 70; 89; 91; 104; 109; 111; 121; 125 (bdsp); 127 (wssp); jul–nov 2012; may 2013; sep 2014, 2016. fomitopsis pinicola (sw.) p. karst. – on logs of pinus and fagus; fq, lf; div 1c; 14 (gosań mt); 31; 39; 46, 48; 68; 70; 104; 110; 113; 114; jul–nov 2012; sep 2014, 2016. galerina vittiformis (fr.) singer – among mosses; div 1a; 1b; aug 2012; sep 2014. ganoderma lipsiense (batsch) g. f. atk. [= g. applanatum (pers.) pat.] – on logs and trunks of fagus; fq, lf, gf; div 13, 14 (gosań mt); 15; 15b; 16; 68; 93; 127, 128a, 128az (wssp); jul–nov 2012; sep 2016. geastrum fimbriatum fr. – on ground; lf; div 89a; oct 2012; r. g. rufescens pers. – on ground; lf; div 11; oct 2012; (leg. m. wilchelm – personal information, 2012); e. gloeoporus dichrous (fr.) bres. – on wood of deciduous tree; lf; div 68; oct 2012; e. gloeophyllum sepiarium (wulfen) p. karst. – on logs of pinus; lf; div 48; 113; 114; sep, nov 2012. gomphidius roseus (fr.) fr. – on ground; div 91; oct 2012; r; ml. gymnopilus penetrans (fr.) murril – on wood; lf; div 2 (sjsp); 13 (gosań mt); 17; 20; 25 (mrsp); 30; 31; 32; 41; 48; 69; 70; 91; 104; 111; 115; aug, oct–nov 2012; sep 2014; ml. gymnopus androsaceus (l.) j. l. mata & r. h. petersen [= setulipes androsaceus (l.) antonín] – on litter; fq, lf, gf; div 1a; 1b; 14 (gosań mt); 16; 17; 25 (mrsp); 30; 31; 39; 41; 62; 69; 70; 89; 91; 93; 111; 128ah; jul–nov 2012; sep 2014; ml. g. confluens (pers.) antonín, halling & noordel. – on litter; lf, gf; div 9; 13, 14 (gosań mt); 16; 48; 68; 89; 111; 125 (bdsp); 128a, 128az (wssp); 128ag; aug, oct– nov 2012; ml. g. dryophilus (bull.) murrill – on litter; fq, cf, lf, gf; div 11 (zcsp); 15b; 16; 48; 68; 69, 71; 89; 91; 105; 111; 125 (bdsp); 127, 128a, 128az (wssp); jul–nov 2012, may 2014; ml, td. 10 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park g. foetidus (sowerby) j. l. mata & r. h. petersen [= marasmiellus foetidus (sowerby) antonín, hlling & noordel.] – on twigs; lf; div 13 (gosań mt); aug 2012; r. g. fusipes (bull.) gray – at the base of stumps of quercus; lf; div 68; 87; 104; 105; aug 2012; sep 2016; ml. g. perforans (hoffm.) antonín & noordel. [= marasmiellus perforans (hoffm.) antonín, hlling & noordel.] – on litter; gf; div 125 (bdsp); oct 2012. g. peronatus (bolton) gray on litter; lf, gf; div 15, 15a; 16; 17; 25 (mrsp); 30; 39; 48; 67; 68; 70; 71; 89; 91; 104; 110; 111; 127 (wssp); aug, oct–nov 2012; sep 2014, 2016; ml. hapalopilus nidulans (fr.) p. karst. – on branches of quercus; cf, lf; div 11 (zcsp); 16; 98; aug–sep 2012; may 2014. hebeloma crustuliniforme (bull.) quél. – on ground; lf, gf; div 11 (zcsp); 98; 116; 125 (bdsp); sep–oct 2012; ml. h. radicosum (bull.) ricken – on roots of fagus; div 98c; sep 2012; i. hericium coralloides (scop.) pers. – on trunks and logs of fagus; lf; div 46d; 91f; 109a (m. walczak – personal information, 2012); oct 2012; sep 2016; v, p. heterobasidion annosum (fr.) bref. – on stumps of pinus; fq, lf, gf; div 1b; 1c; 13, 14 (gosań mt); 25 (mrsp); 31; 41; 48; 70; 91; 110; 111; 114; 115; 125 (bdsp); aug–nov 2012; sep 2014. hydropus subalpinus (höhn.) singer – on twigs; lf, gf; div 1c; 13, 14 (gosań mt); 15; 15b; 16; 17; 30; 31; 32; 91; 104; 105; 125 (bdsp); 127 (wssp); aug, oct 2012; r. hygrophoropsis aurantiaca (wulfen) maire – on ground; fq; div 1a; 1b; 70; aug 2012; sep 2014; ml. hygrophorus eburneus (bull.) fr. – on ground; lf; div 48; 68; 89; 91; 111; 113; sep– nov 2012; ml. h. hypothejus (fr.) fr. – on ground; div 2g,n; 111; oct–nov 2012; i, ml. hymenochaete rubiginosa (dicks.) lév. – on stumps of quercus; lf, gf; div 25 (mrsp); 68; 125 (bdsp); 127 (wssp); 128ay; aug, oct–nov 2012; sep 2016. hyphoderma setigerum (fr.) donk – on wood; lf; div 11, 12 (zcsp); 25 (mrsp); 68; 104; 105; aug, oct–nov 2012; sep 2016. hyphodontia nespori (bres.) j. erikss. & hjortstam – on branches; lf; div 1c; 13, 14 (gosań mt); aug 2012. hypholoma capnoides (fr.) p. kumm. [= psilocybe capnoides (fr.) noordel.] – on wood; lf; div 31; 121; oct 2012. h. fasciculare (huds.) p. kumm. [= p. fascicularis (huds.) kühner] – on wood; fq, lf, gf; div 1b; 2g,n; 2 (sjsp); 15; 25 (mrsp); 31; 39; 40; 48; 68; 69; 91; 97; 98; 104; 105; 111; 113; 114; 116; 125 (bdsp); 128a, 128az (wssp); jul–nov 2012; sep 2014, 2016; ml. h. lateritium (schaeff.) p. kumm. [= p. lateritia (schaeff.) noordel.] – on wood; lf; div 31; 89; oct 2012; ml. 11 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park imleria badia (fr.) vizzini [= xerocomus badius (fr.) e.-j. gilbert] – on ground; lf; div 46; 68; 70; 91; 94; 97; 110; sep–nov 2012; sep 2014, 2016; ml, ms. inocybe asterospora quél. – on ground; lf, gf; div 13, 14 (gosań mt); 16; 32; 70; 71; 91; 94; 105; 111; 115; 121; aug–oct 2012. i. dulcamara (pers.) p. kumm. – on ground; lf; div 105; aug 2012; ml. i. rimosa (bull.) p. kumm. [= i. fastigiata (schaeff.) quél.] – on ground; lf; div 15b; aug 2012; ml. i. geophylla (sowerby) p. kumm. var. geophylla – on ground; lf, gf; div 20; 48; 71; 91; 110; 111; 125 (bdsp); oct–nov 2012; ml. i. geophylla var. lilacina (peck) gillet – on ground; lf, gf; div 116; 125 (bdsp); oct 2012; ml. i. griseolilacina j. e. lange – on ground; lf; div 116b; oct 2012; r. i. hirtella bres. – na ziemi; lf; div 68; 105; aug, oct 2012; ml. i. lacera (fr.) p. kumm. – on ground; lf; div 1a; 1b; 15; 91; 111; 114; 121; aug–oct 2012. i. lanuginosa (bull.) p. kumm. – on ground; lf, gf; oddz. 16; 17; 105; 111; 121; oct 2012; ml. i. maculata boud. – on ground; div 105; aug 2012. i. praetervisa quél. – on ground; gf; div 39; 127 (wssp); aug 2012. innonotus nodulosus (fr.) p. karst. – on trunks of fagus; lf, gf; div 15b; 31; 46; 68; 125 (bdsp); aug, oct 2012; sep 2016. i. obliquus (ach. ex pers.) pilát – on trunks of betula; div 1c; aug 2012; r, p. ischnoderma resinosum (schrad.) p. karst. – on logs of fagus; lf, gf; div 89a; 93l; 125 (bdsp); oct 2012; v. kuehneromyces mutabilis (schaeff.) singer & a. h. sm. – on wood of deciduous trees; fq, lf; div 2 (sjsp); 31; 39; 69; nov 2012; sep 2014; ml. laccaria amethystea (bull.) murrill – on ground; lf, gf; div 2 (sjsp); 8; 11, 12 (zcsp); 25 (mrsp); 30; 32; 39; 48; 68; 69; 70; 91; 109; 110; 111; 114; 115; 116; 121; 125 (bdsp); 127 (wssp); aug–nov 2012; sep 2014; ml. l. laccata (scop.) cooke – on ground; lf, gf; div 2 (sjsp); 11 (zcsp); 25 (mrsp); 32; 48; 125 (bdsp); oct–nov 2012; ml. l. proxima (boud.) pat. – on ground; div 91; oct 2012; ml. l. tortilis (bolton) cooke – on ground; gf; div 25 (mrsp); 125 (bdsp); oct–nov 2012. lactarius aurantiacus (pers.) gray – on ground; div 20; nov 2012; ml. l. blennius (fr.) fr. – on ground; lf, gf; div 2 (sjsp); 11 (zcsp); 68; 89; 91; 109; 111; 121; 125 (bdsp); oct–nov 2012; ml. 12 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park l. deliciosus (l.) gray – on ground; lf; div 1a; 1b; 91; 93; 94; 97; aug–oct 2012; ml. l. fluens boud. – on ground; lf; div 20; 25 (mrsp); 32; 48; 91; 94; 98; 104; 114; aug–nov 2012. l. necator (bull.) pers. – on ground; fq; div 69; sep 2014. l. quietus (fr.) fr. – on ground; fq, lf, gf; div 1c; 14 (gosań mt); 16; 39; 40; 68, 69; 94; 104; 109; 114; 125 (bdsp); 127, 128a, 128az (wssp); jul–oct 2012; sep 2014; ml. l. rufus (scop.) fr. – on ground; lf; div 9; 14 (gosań mt); 15, 15a; 39; 40; 91; 94; 104; 105; 111; aug–oct 2012; ml. l. tabidus fr. [= l. thejogalus (bull.) gray ss. neuhoff ] – on ground; fq, lf, gf; div 1c; 8; 9; 14 (gosań mt); 15, 15a; 31; 39; 69; 70; 91; 93; 104; 109; 111; 128a, 128az (wssp); aug–oct 2012; sep 2014. l. vellereus (fr.) fr. – on ground; lf, gf; div 2 (sjsp); 25 (mrsp); 30; 31; 32; 68; 71; 89; 91; 93; 94; 97; 98; 104; 111; 113; 114; 115; 116; 125 (bdsp); 127 (wssp); aug–nov 2012; ml. l. vietus (fr.) fr. – on ground; lf; div 110; oct 2012. l. volemus (fr.) fr. – on ground; div 91; oct 2012. laetiporus sulphureus (bull.) murrill – on trunk of quercus; lf; div 46; sep 2016; ml. leccinum scabrum (bull.) gray – on ground; div 110; oct 2012; ml, ms. lenzites betulina (l.) fr. – on logs of fagus; gf; div 125 (bdsp); oct 2012. lepiota cristata (bolton) p. kumm. – on ground; lf, gf; div 116; 125 (bdsp); oct 2012; ml. lepista flaccida (sowerby) pat. – on litter; lf, gf; div 14 (gosań mt); 25 (mrsp); 48; 125 (bdsp); aug, oct–nov 2012. l. nuda (bull.) cooke – on ground; lf, gf; div 68; 89; 116; 125 (bdsp); oct 2012; ml. leratiomyces squamosus (pers.) bridge & spooner [= psilocybe squamosa (pers.) p. d. orton] – on ground and litter; lf, gf; div 31; 71; 91; 109a; 110; 111; 114; 121; 125 (bdsp); sep–oct 2012; i; ml. lycoperdon echinatum pers. – on ground; div 6; oct 2012; (leg. m. wilchelm – personal information, 2012); r. l. foetidum bonord. (= l. nigrescens wahlenb.) – on ground; div 69; sep 2014. l. perlatum pers. – on ground; fq, lf, gf; oddz. 2 (sjsp); 15a; 25 (mrsp); 30; 68; 70; 71; 89; 91; 93; 109; 111; 115; 125 (bdsp); aug–nov 2012; sep 2014, 2016; ml. l. pyriforme schaeff. – on wood and old basidiocarps of fomes fomentarius; lf, gf; div 48; 115, 125 (bdsp); 127 (wssp); jul–nov 2012; ml. l. umbrinum pers. – on ground; lf; oddz. 2g,n; div 2 (sjsp); nov 2012. 13 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park macrolepiota procera (scop.) singer – on ground; div 46; sep 2016; ml. macrotyphula contorta (holmsk.) rauschert [= clavariodelphus fistulosus (holmsk.) corner] – on twigs; lf; div 2 (sjsp); 25g (mrsp); nov 2012; r. marasmiellus ramealis (bull.) singer – on twigs of fagus; gf; div 127 (wssp); aug 2012; ml. marasmius epiphyllus (pers.) fr. – on litter; lf, gf; div 16; 128a, 128az (wssp); aug 2012. m. rotula (scop.) fr. – on twigs; lf, gf; div 13, 14 (gosań mt); 15b; 16; 104; 127 (wssp); aug 2012; ml. m. torquescens quél. – on twigs and litter; gf; div 127 (wssp); aug 2012; ml. megacollybia platyphylla (pers.) kotl. & pouzar – on wood of deciduous trees; fq, lf, gf; div 2 (sjsp); 14 (gosań mt); 15; 15b; 16; 71; 67; 68; 69; 70; 89; 91; 104; 111; 114; 127, 128a, 128az (wssp); jul–nov 2012; sep 2014, 2016; ml. meripilus giganteus (pers.) p. karst. – on logs of fagus; lf, gf; div 109a (m. walczak – personal information, 2012); 116b; 128ay; aug, oct 2012. mutinus caninus (huds.) fr. – on ground; lf, gf; div 13d (gosań mt); 15o, m, 15ac; 15bj (kawcza mt); 110; 127 (wssp); aug, oct 2012; sf. mycena capillaris p. karst. – on leaves of fagus; lf; div 115; oct 2012; ml. m. crocata (schrad.) p. kumm. – on litter; lf; div 2j (sjsp); 109a; 115; 121; oct–nov 2012; r. m. epipterygia (scop.) gray var. epipterygia – on ground and among mosses; lf; div 2 (sjsp); 30; 91; 111; 121; oct–nov 2012; ml. m. filopes (bull.) p. kumm. – on twigs; lf; div 31; 89; 91; 111; oct 2012; ml. m. galericulata (scop.) gray – on stumps of deciduous trees; lf, gf; div 2 (sjsp); 8; 13 (gosań mt); 16; 25 (mrsp); 46; 68; 93; 104; 109; 121; 125 (bdsp); aug–nov 2012; sep 2016; ml. m. galopus (pers.) p. kumm. – on litter; fq, lf, gf; div 1a; 1b; 1c; 2 (sjsp); 8; 9; 20; 25 (mrsp); 30; 31; 32; 40; 41; 48; 69; 70; 89; 91; 104; 110; 111; 115; 116; 121; 125 (bdsp); 128a, 128az (wssp); 128ah; jul–nov 2012; sep 2014; ml. m. haematopus (pers.) p. kumm. var. haematopus – on stumps and logs of deciduous trees; lf, gf; div 1c; 2 (sjsp); 13 (gosań mt); 15; 30; 39; 70; 89; 91; 104; 110; 111; 116; 121; 125 (bdsp); aug, oct–nov 2012. m. inclinata (fr.) quél. – on stumps of quercus; lf; div 25 (mrsp); 48; 68; oct–nov 2012; ml. m. pelianthina (fr.) quél. – on litter; lf, gf; div 48; 125 (bdsp); oct–nov 2012; i, ml. m. polygramma (bull.) gray – at the base of trunks of deciduous trees; lf, gf; div 115; 125 (bdsp); oct 2012; ml. m. pura (pers.) p. kumm. – on ground and litter; fq, lf, gf; div 1a; 1b; 1c; 2g,n; 2 (sjsp); 13, 14 (gosań mt); 17; 20; 25 (mrsp); 48; 68; 70; 89; 91; 98; 109; 111; 113; 114; 125 (bdsp); 128ag; aug–nov 2012; sep 2014; ml. 14 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park m. sanguinolenta (alb. & schwein.: fr.) p. kumm. – on litter; lf, gf; div 1c; 13, 14 (gosań mt); 15a; 15b; 16; 30; 40; 41; 70; 89; 115; 127, 128a, 128az (wssp); aug, oct 2012; sep 2014; ml. m. stipata maas geest. & schwöbel. – on branches and logs of pinus; lf; div 2g,n; 30; 31; nov 2012. m. stylobates (pers.) p. kumm. – on leaves of fagus; lf; div 1c; 104; aug 2012; ml. m. vitilis (fr.) quél. – on litter; lf, gf; div 1a; 1b; 2 (sjsp); 11, 12 (zcsp); 15a, 48; 121; 125 (bdsp); 128ag; aug, oct–nov 2012. m. vulgaris (pers.) p. kumm. – on needles, among mosses; div 91; 111; oct 2012; ml. m. zephirus (fr.) p. kumm. – on ground and litter; fq, lf, gf; div 15a; 25 (mrsp); 31; 32; 68; 69; 70; 91; 110; 111; 115; 125 (bdsp); 128ah; aug, oct–nov 2012; sep 2014. mycetinis alliaceus (jacq.) earle [= marasmius alliaceus (jacq.) fr.] – on wood of fagus; fq, lf, gf; div 2 (sjsp); 11, 12 (zcsp); 13, 14 (gosań mt); 15, 15a; 15b; 16; 25 (mrsp); 30; 31; 32; 40; 68; 69; 70; 71; 89; 97; 98; 104; 109; 114; 115; 116; 121; 125 (bdsp); 127, 128a, 128az (wssp); aug–nov 2012; sep 2014; ml. oudemansiella mucida (schrad.) höhn. – on logs and stumps of fagus; lf, gf; div 2 (sjsp); 11, 12 (zcsp); 31; 48; 68; 89; 71; 91; 94; 97; 104; 111; 113; 116; 125 (bdsp); aug–nov 2012; zcju, ml. panellus serotinus (pers.) kühner – on logs of fagus; lf, gf; div 2 (sjsp); 48; 125 (bdsp); oct–nov 2012. p. stipticus (bull.) p. karst. – on stumps of quercus; lf, gf; div 2 (sjsp); 48; 71; 125 (bdsp); oct–nov 2012. paxillus involutus (batsch) fr. – on ground; fq, lf; div 30; 31; 68; 70; 91; 94; 111; sep–oct 2012; sep 2014; ml. peniophora cinerea (pers.) cooke – on branches; fq, lf; div 15; 16; 69; 70; 111; aug, oct 2012; sep 2014. p. quercina (pers.) cooke – on branches of quercus; cf, lf; div 2 (sjsp); 11 (zcsp); 16; 25 (mrsp); 31; 40; 46; 68; 97; 98; 114; jul–nov 2012; may 2014; sep 2016. phaeolus schweinizii (fr.) pat. – on roots of pinus; div 15a; aug 2012. phallus impudicus l. – on ground; lf, gf; div 11, 12 (zcsp); 14 (gosań mt); 15, 15a; 31; 48; 70; 91; 94; 104; 110; 111; 125 (bdsp); 128a, 128az (wssp); aug–nov 2012; sep 2014; ml, as. phanerochaete laevis (fr.) j. erikss. & ryvarden – on wood of fagus; lf; div 13, 14 (gosań mt); 15, 15ac; 15b; 16; 17; 111; aug, oct 2012. ph. sanguinea (fr.) pouzar – on twigs; fq; div 40; 69; aug 2012; sep 2014. ph. velutina (dc.) p. karst. – on litter and branches; gf; div 127 (wssp); aug 2012. phellinus ferruginosus (schrad.) pat. – on branches of quercus; fq, lf; div 48; 68; 70; 97; 98; 104; 113; aug–nov 2012; sep 2014, 2016. 15 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park ph. hippophaëcola h. jahn – on branches of hippophäe rhamnoides; wapnica (n lakeside of turkusowe lake); may 2013; ww. ph. robustus (p. karst.) bourdot & galzin – on wood of quercus; lf, gf; div 46; 68; 105; 128a, 128az (wssp); aug 2012; sep 2016. phlebia radiata fr. – on logs of fagus; lf, gf; div 25 (mrsp); 89; 109; 125 (bdsp); oct–nov 2012. ph. tremellosa (schrad.) nakasone & burds. – on logs of fagus; lf, gf; div 25 (mrsp); 31; 48; 70, 125 (bdsp); oct–nov 2012; sep 2014. pholiota aurivella (batsch) p. kumm. – on logs of fagus; gf; div 89; 104; aug, oct 2012. ph. flammans (batsch) p. kumm. – on logs of pinus; fq; div 70; 94; sep 2012, 2014. ph. lenta (pers.) singer – on litter; lf, gf; div 109; 115; 116; 125 (bdsp); oct 2012. piptoporus betulinus (bull.) p. karst. – on wood of betula; lf; div 1b; 1c; 91; 70; 110; 111; aug–nov 2012; sep 2014. pluteus cervinus (schaeff.) p. kumm. – on wood of deciduous trees; lf, gf; div 2 (sjsp); 11, 12 (zcsp); 13 (gosań mt); 30; 68; 89; 69; 71; 104; 109; 116; 121; 125 (bdsp); jul–nov 2012; sep 2014; ml. p. romellii (britzelm.) sacc. – on logs; lf, gf; div 89a; 125 (bdsp); oct 2012. p. salicinus (pers.) p. kumm. – on logs of fagus; lf, gf; div 14 (gosań mt); 89a; 128a, 128az (wssp); aug, oct 2012; sep 2014. p. umbrosus (pers.) p. kumm. – on log of fagus; lf; div 89a; oct 2012. polyporus ciliatus fr. – on branches; fq, lf, gf; div 48; 70; 89; 125 (bdsp); oct–nov 2012; sep 2014. p. tuberaster (jacq. ex pers.) fr. – on branches; fq, lf, gf; div 1c; 15; 17; 69; 91; 104; 105; 111; 127 (wssp); aug, oct 2012; sep 2014; r. p. varius (pers.) fr. – on branches of fagus; cf, fq, lf, gf; div 1c; 2 (sjsp); 11, 12 (zcsp); 13, 14 (gosań mt); 15, 15a; 15b; 16; 17; 30; 31; 40; 46; 67; 68; 69; 93; 94; 97; 104; 113; 114; 127, 128a, 128az (wssp); 128ah; jul–nov 2012; my, sep 2014; sep 2016; ml. porodaedalea pini (brot.) murrill [= phellinus pini (brot.) a. ames] – on trunks and logs of pinus; lf, gf; div 11j (zcsp); 46d; 68g; 125 (bdsp); oct 2012; may 2013; sep 2016; r. postia caesia (schrad.) p. karst. [= oligoporus caesius (schrad.) gilb. & ryvarden] – on wood of pinus; lf, gf; div 2 (sjsp); 31; 48; 89; 91; 111; 125 (bdsp); 128a, 128az (wssp); aug, oct–nov 2012. p. ptychogaster (f. ludw.) vesterh. [= o. ptychogaster (f. ludw.) falck & o. falck] – on logs of pinus; div 115c; oct 2012; r. p. stiptica (pers.) jülich [= o. stipticus (pers.) gilb. & ryvarden] – on wood of pinus; lf, gf; div 15a; 17; 41; 91; 111; 128ah; aug, oct 2012. 16 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park psathyrella candolleana (fr.) maire – on ground and litter; lf, gf; div 15b; 25 (mrsp); 104; 128a, 128az (wssp); aug, nov 2012. p. friesii kits van wav. [= p. fibrillosa (pers.) maire] – on litter; lf; div 2 (sjsp); 30; 91; 111; 109; oct–nov 2012. p. murcida (fr.) kits van wav. – on litter; lf; div 115b; oct 2012. p. piluliformis (bull.) p. d. orton – on wood of fagus and quercus; fq, lf, gf; div 2 (sjsp); 20; 25 (mrsp); 30; 31; 48; 68; 69; 89; 109; 110; 115; 116; 121; 125 (bdsp); oct–nov 2012; sep 2014. p. prona (fr.) gillet – on ground; gf; div 125 (bdsp); oct 2012. pseudoclitocybe cyathiformis (bull.) singer – on ground; gf; div 125 (bdsp); oct 2012. pseudohydnum gelatinosum (scop.) p. karst. – on stumps of pinus; lf, gf; div 48; 125 (bdsp); oct–nov 2012. pseudomerulius aureus (fr.) jülich – on logs of pinus; fq; div 69; 111; sep 2012, 2014; r. radulomyces molaris (chaillet ex fr.) m. p. christ. – on branches of quercus; div 25 (mrsp); 39; 98; aug–nov 2012. ramaria stricta (pers.) quél. – on ground; lf; div 2 (sjsp); 15a; 91; 114; 111; aug– nov 2012. resupinatus trichotis (pers.) singer – on branch; div 1c; aug 2012. rhizopogon luteolus fr. & nordholm – on ground; fq; div 70m (n lakeside of czajcze lake); sep 2014; ml. rhodocollybia butyracea (bull.) lennox f. butyracea – on ground; fq, lf, gf; div 2g,n; 2 (sjsp); 11, 12 (zcsp); 25 (mrsp); 48; 68; 69; 71; 91; 109; 110; 111; 121; 125 (bdsp); 128ag; aug, oct–nov 2012; sep 2014. rh. maculata (alb. & schwein.) singer – on ground; fq, lf; div 48; 70; 89; oct–nov 2012; sep 2014; ml. rhodonia placenta (fr.) niemelä, k. h. larss. & schigel [= oligoporus placentus (fr.) gilb. & ryvarden] – on log of pinus; div 39 (road to bison show reserve); aug 2012; e. rickenella fibula (bull.) raithelh. – among mosses; lf; div 1a; 1b; 8; 9; 13 (gosań mt); 32; 89; aug, oct x 2012; ml. r. swartzii (fr.) kuyper – on ground and among mosses; div 1a; 1b; aug 2012. roridomyces roridus (fr.) rexer [= m. rorida (fr.) quél.] – on litter and among mosses; div 64; sep 2014; ml. russula amoenolens romagn. – on ground; div 98c; sep 2012; r. r. atropurpurea (krombh.) britzelm. (= r. undulata volen.) – on ground; gf; div 125 (bdsp); oct 2012; ml. r. betularum hora – on ground; fq; div 1a; 1b; 69; 91; aug, oct 2012; sep 2014. 17 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park r. caerulea fr. – on ground; fq; div 70i; sep 2014; r. r. claroflava grove – on ground; lf; div 16; aug 2012. r. cyanoxantha (schaeff.) fr. – on ground; lf, gf; div 15; 15b; 16; 31; 48; 71; 93; 97; 104; 111; 113; 121; 128a, 128az (wssp); aug–nov 2012; ml. r. decolorans (fr.) fr. – on ground; div 41; 111; aug, oct 2012; ml. r. emetica (schaeff.) pers. – on ground; lf; div 1a; 1b; 1c; 2g,n; 9; 14 (gosań mt); 15; 16; 17; 31; 32; 40; 62; 68; 91; 97; 111; 113; 114; 121; aug–nov 2012; ml. r. fellea (fr.) fr. – on ground; lf, gf; div 2g,n; 2 (sjsp); 9; 11, 12 (zcsp); 14 (gosań mt); 15; 15b; 16; 25 (mrsp); 48; 68; 91; 93; 94; 97; 105; 109; 111; 113; 114; 125 (bdsp); 127, 128a, 128az (wssp); aug–nov 2012; ml. r. foetens pers. – on ground; lf; div 89; 104; 113; aug–sep 2012; ml. r. fragilis fr. – on ground; fq, lf, gf; div 69; 71; 94; 98; 127 (wssp); aug–oct 2012; sep 2014; ml. r. lutea (huds.) gray – on ground; lf, gf; div 71; 91; 111; 114; 121; sep–oct 2012; ml. r. mairei singer – on ground; lf gf; div 11 (zcsp); 15; 25 (mrsp); 31; 48; 93; 94; 97; 98; 104; 114; 115; 125 (bdsp); aug–nov 2012. r. nigricans fr. – on ground, lf, gf; div 8; 13, 14 (gosań mt); 16; 30; 46, 67; 71; 89; 91; 93; 94; 97; 98; 104; 105; 111; 113; 115; 121; 125 (bdsp); 127 (wssp); aug–nov 2012; sep 2016; ml. r. ochroleuca pers. – on ground; lf, gf; div 2 (sjsp); 13 (gosań mt); 25 (mrsp); 30; 31; 32; 68; 71; 89; 91; 111; 113; 125 (bdsp); aug–nov 2012. r. paludosa britzelm. – on ground; div 1b; aug 2012. r. risigallina (batsch) sacc. – on ground; lf; div 16; aug 2012; ml. r. sanguinea (bull.) fr. – on ground; div 1a; 1b; aug 2012; ml. *r. torulosa bres. – on ground; div 1a; 1b; aug 2012, sep 2013–2014, oct 2015; mst. r. xerampelina (schaeff.) fr. – on ground; lf; div 1a; 1b; 48; 91; 93; 94; 111; aug– nov 2012. schizophyllum commune fr. – on wood; zielonka hill; may 2013; ml. schizopora paradoxa (schrad.) donk [= hyphodontia paradoxa (schrad.) langer & vesterholt] – on wood; fq, lf, gf; div 1c; 2 (sjsp); 11, 12 (zcsp); 13, 14 (gosań mt); 15; 15b; 16; 30; 40; 68; 89; 98; 104; 105; 113; 128ag; 128a, 128az (wssp); jul–nov 2012; sep 2016. scleroderma citrinum pers. – on ground; fq; div 48; 70; nov 2012; sep 2014; ml. s. verrucosum (bull.): pers. – on ground; lf, gf; div 8; 9; 25 (mrsp); 31; 32; 39; 41; 68; 91; 97; 98; 104; 111; 113; 114; 121; aug–nov 2012; ml. sparassis crispa (wulfen) fr. – at the base of pinus; lf; div 11h (zcsp); 46d; 113; sep–oct 2012; sep 2016; r; ml. 18 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park sphaerobolus stellatus tode – on wood; gf; div 125 (bdsp); oct 2012. stereum hirsutum (willd.) pers. – on wood; fq, lf, gf; div 1c; 2 (sjsp); 13, 14 (gosań mt); 15; 15a; 15b; 16; 25 (mrsp); 30; 32; 46; 48; 62; 68; 70; 89; 104; 114; 127, 128a, 128az (wssp); jul–nov 2012; sep 2014, 2016; ml. s. rugosum pers. – on wood; fq, lf, gf; div 1b; 1c; 11 (zcsp); 13 (gosań mt); 15b; 16; 25 (mrsp); 48; 62; 91; 104; 105; 111; 121; 125 (bdsp); 128a, 128az (wssp); jul–nov 2012. s. sanguinolentum (alb. & schwein.) fr. – on branches of pinus; fq, lf; div 1c; 9; 40; 41; 62; 69; 70; 91; 104; 111; 121; 128ah; jul–nov 2012; sep 2014. s. subtomentosum pouzar – on logs; lf, gf; div 2 (sjsp); 13 (gosań mt); 125 (bdsp); 127 (wssp); aug, nov 2012. strobilurus stephanocystis (kühner & romagn. ex hora) singer – on cone of pinus; zielonka hill; may 2013–2014; ml. stropharia aeruginosa (curtis) quél. [= psilocybe aeruginosa (curst.) noordel.] – on ground and wood; fq, lf, gf; div 2g, n; 2 (sjsp); 25 (mrsp); 31; 48; 68; 70; 109; 110; 116; 125 (bdsp); oct–nov 2012; sep 2014; ml. s. semiglobata (batsch) quél. [= psilocybe semiglobata (batsch) noordel.] – on droppings; div 115; oct 2012. suillus bovinus (l.) roussel – on ground; div 91; oct 2012; ml, ms. s. luteus (l.) roussel – on ground; div 1b; sep 2014; ml, ms. s. grevillei (klotzsh) singer – on ground; lf; div 113; 114; sep 2012; ml. tapinella atrotomentosa (batsch) šutara [= paxillus atrotomentosus (batsch) fr.] – on logs and stumps of pinus; div 30; 39; 41; aug, oct 2012; ml. t. panuoides (fr.) e.-j. gilbert [= p. panuoides (fr.) fr.] – on logs of pinus; lf; div 15a; 111; aug, oct 2012. thelephora terrestris ehr. – on ground and wood; fq; div 1a; 1b; 9; 31; 39; 70; aug 2012; sep 2014; ml. trametes gibbosa (pers.) fr. – on stumps and logs of fagus; lf, gf; div 13, 14 (gosań mt); 48; 94; 110; 113; 114; 125 (bdsp); jul–nov 2012. t. hirsuta (wulfen) pilát – on wood; lf, gf; div 1c; 8; 15a; 39; 40; 48; 127, 128a, 128az (wssp); jul–nov 2012. t. ochracea (pers.) gilb. & ryvarden – on wood; gf; div 111; 121; 125 (bdsp); oct 2012. t. pubescens (schumach.) pilát – on wood; gf; div 127 (wssp); aug 2012; r. t. versicolor (l.) lloyd – on wood; lf; div 89; 91; 97; sep–oct 2012; ml. tremella encephala pers. – on branches of pinus and basidiocarps of stereum sanguinolentum; div 9; 128ah; aug 2012. t. mesenterica (schaeff.) retz. – on twigs of quercus; fq, lf; div 68; 69; oct 2012; sep 2014; ml. 19 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park trichaptum abietinum (pers. ex j. f. gmel.) ryvarden – on wood of pinus; fq, lf, gf; div 1b; 1c; 2g,n; 8; 9; 15; 31; 39; 40; 41; 46; 62; 68; 70; 97; 125 (bdsp); 128ah; jul–nov 2012; sep 2014, 2016; ml. t. fuscoviolaceum (ehrenb.) ryvarden – on logs of pinus; lf; div 1a; 1b; 41; 104; aug 2012. tricholoma lascivum (fr.) gillet – on ground; gf; div 125 (bdsp); oct 2012. t. scalpturatum (fr.) quél. – on ground; lf; div 2 (sjsp); 20; nov 2012. t. sulphureum (bull.) p. kumm. – on ground; lf; div 2 (sjsp); 116; oct–nov 2012; ml. t. terreum (schaeff.) p. kumm. – on ground; gf; div 1a; 1b; 2g,n; 125 (bdsp); oct– nov 2012; ml. t. virgatum (fr.) p. kumm. – on ground; lf, gf; div 71; 125 (bdsp); oct 2012. tricholomopsis rutilans (schaeff.) singer – on stumps of pinus; lf; div 15; 31; 70; aug, oct 2012; sep 2014; ml. tubaria furfuracea (pers.) gillet – on ground; lf, gf; div 109; oct 2012. tylopilus felleus (bull.) p. karst. – on ground; lf; div 9; 16; 17; 39; 40; 46; 88; 105; aug 2012; sep 2016; ml, ms. volvariella bombycina (schaeff.) singer – at the base of fagus; lf; div 46d; sep 2016; r. vuilleminia comedens (nees) mraire – on branches of quercus; lf, gf; div 14 (gosań mt); 39; 41; 46; 48; 68; 89; 98; 114; 127 (wssp); jul–nov 2012; may 2013; sep 2016. xerocomellus chrysenteron (bull.) šutara [= xerocomus pascuus (pers.) e.-j. gilbert] – on ground; lf, gf; div 2 (sjsp); 11, 12 (zcsp); 20; 25 (mrsp); 30; 31; 32; 48; 68; 70; 71; 91; 97; 98; 111; 113; 114; 115; 121; 125 (bdsp); 127 (wssp); jul–nov 2012; sep 2014; ml, ms. xerocomus subtomentosus (l.) quél. – on ground; lf; div 1c, 46; aug 2012; sep 2016; ml, ms. xeromphalina caulicinalis (fr.) kühner & maire (= x. fellea maire & malençon) – on ground and needles; lf; div 8; 9; 13 (gosań mt); 70; 91; 111; aug, oct 2012; sep 2014. xerula radicata (relhan) dörfelt – on roots of fagus; lf, gf; div 13, 14 (gosań mt); 15, 15a; 15b; 16; 48; 67, 68, 89; 94; 97; 104; 105; 110; 114; 116; 125 (bdsp); 127 (wssp); jul–nov 2012; sep 2016; ar. xylobolus frustulatus (pers.) boidin – on old logs of quercus; cf, lf; div 11 (zcsp); 46d, 68g; may 2014; sep 2016; v. discussion in total, in wnp 476 fungal taxa have been identified so far, which constitutes approximately 13% of all macromycetes known from poland [40–42]. over one third (i.e., 179 species) of them, had not been reported until now from investigated area. 20 of 22© the author(s) 2017 published by polish botanical society acta mycol 51(2):1089 stasińska and sotek / macrofungi of wolin national park the occurrence of 143 species given by, e.g., lisiewska [2], stier [18], skirgiełło [21], ławrynowicz [23], and friedrich [27] has been confirmed. although conducted studies focused on selected phytocoenoses, they revealed a great diversity in the park mycobiota. the number of macrofungi taxa found in wnp is higher than the number of fungi reported from other national parks in pomerania. almost 100 fewer taxa were given from the drawieński national park (379) (stefaniak 2013, cited in [43]). also, a lower number of fungal species in comparison with wnp, was recorded in np bory tucholskie (413) [8] and in the słowiński np (429) [3]. as a rule, national parks in other regions of poland, e.g., kampinowski np (1533) [43], bieszczadzki np (1054) (kujawa et al. 2011, cited in [43]), have a greatest variety and abundance of fungi. however, it should be noted that the studies took place at different intensity, on the objects of different size, in different ecosystems and plant associations of varying diversity. mycological observations conducted in season 2012 and sporadic observations in the following 4 years, as well as available literature data may constitute for further detailed studies of macrofungi of the wolin national park. fluctuations and periodicity of the appearance of fruiting bodies do not allow for detailed mycological analysis and drawing far-reaching conclusions. the number of fungi in the wnp is likely to be higher and further studies may result in discovering other rare and endangered taxa that had not been reported earlier. acknowledgments the authors would like to thank mgr inż. mateusz walczak and dr marcin wilhelm for providing their unpublished mycological data. we also thank the staff of wolin national park for their help in field research and mgr jerzy prajs for the drawing used in this study. references 1. ustawa z dnia 6 lipca 2001 r. o zachowaniu narodowego charakteru strategicznych zasobów naturalnych kraju. journal of laws of the republic of poland (dziennik ustaw), 11 september 2001, no. 97, item 1051. 2. lisiewska m. grzyby wyższe wolińskiego parku narodowego. acta mycol. 1966;2:25–77. http://dx.doi.org/10.5586/am.1966.005 3. bujakiewicz a, lisiewska m. mikoflora zbiorowisk roślinnych słowińskiego parku narodowego. badania fizjograficzne nad polską zachodnią. seria b – botanika. 1983;34:49–77. 4. wojewoda w. macromycetes ojcowskiego parku narodowego. i. flora. acta mycol. 1974;10:181–265. http://dx.doi.org/10.5586/am.1974.007 5. łuszczyński j. grzyby wielkoowocnikowe. in: ciesliński s, kowalkowski a, editors. świętokrzyski park narodowy. przyroda, gospodarka, kultura. bodzentyn: świętokrzyski park narodowy; 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(biodiversity of poland; vol 9). 43. karasiński d, kujawa a, gierczyk b, ślusarczyk t, szczepkowski a. grzyby wielkoowocnikowe kampinowskiego parku narodowego. izabelin: petit s.k.; 2015. http://dx.doi.org/10.1127/nova_hedwigia/2016/0341 abstract introduction methods results list of species ascomycota basidiomycota discussion acknowledgments references 2017-01-23t12:02:50+0000 piotr otręba analysis of virulence and genetic variability of alternaria alternata associated with leaf spot disease in potato plants in iran 1 of 9published by polish botanical society acta mycologica original research paper analysis of virulence and genetic variability of alternaria alternata associated with leaf spot disease in potato plants in iran mehdi nasr esfahani* plant protection research department, isfahan agricultural and natural resources research and education center (areeo), isfahan, iran * email: mne2011@gmail.com abstract leaf spot disease in potato is caused by alternaria alternata (fr.) keissler, an opportunistic pathogen that infests many agricultural crops worldwide in the field and during postharvest storage of vegetables and fruits. alternaria alternata is associated with leaf spot disease in potato in iran. thus, there is a need to investigate the virulence and genetic variability of iranian a. alternata isolates to facilitate the development of appropriate management strategies. in the present study, we analyzed a total of 28 isolates obtained from the main potato-growing regions of iran, including the ardebil, hamedan, isfahan, and fars provinces. the pathogens were characterized based on sequence analysis of the genes encoding glyceraldehyde3-phosphate dehydrogenase (gpd), plasma membrane atpase, alternaria allergen a 1 (alt a1), calmodulin, and actin. in addition, random amplified polymorphic dna (rapd), intersimple sequence repeat (issr), and virulence studies were performed. phylogenetic analysis of the combined dataset indicated that the five representative isolates were grouped with the subcluster comprising a. alternata. rapd and issr analyses clustered the 28 a. alternata isolates into different groups with no correlation with their corresponding geographical origins. results of the pathogenicity assay indicated that all a. alternata isolates were pathogenic against potato. however, the a. alternata isolates showed high variability in terms of virulence. keywords alternaria alternata; issr; rapd; solanum tuberosum l.; virulence variability introduction alternaria alternata (fr.) keissler has been isolated from a wide range of crops, including fruits and vegetables, nuts, and cereals [1]. a. alternata is a common saprobe found in many plants and other substrata worldwide and is an opportunistic pathogen that infects many agricultural crops in the field and during postharvest storage of vegetables and fruits. in potato, the a. alternata fungus causes brown necrotic lesions in foliage and black pit [2]. previous reports have shown that a. alternata can destroy more than 20% of potato production [3,4]. in iran, two species, namely, a. alternata and a. solani (ellis & martin), have been identified as the causal agents of leaf spot disease in potato, with a. alternata being the dominant species [5]. in recent years, multigene phylogeny has been widely employed for identification and characterization of alternaria species [6,7]. molecular approaches based on sequence analysis of gene fragments encoding glyceraldehyde-3-phosphate dehydrogenase (gpd), plasma membrane atpase, alternaria allergen a 1 (alt a1), calmodulin, and actin have robustly defined the monophyly of alternaria in the ascomycete family pleosporaceae [5]. the efficacy of control strategies on the plant pathogen populations have been hampered by limited information on genetic variability [6,8,9]. the most commonly doi: 10.5586/am.1105 publication history received: 2018-02-15 accepted: 2018-05-18 published: 2018-06-29 handling editor wojciech pusz, faculty of environmental and life sciences, wrocław university of environmental and life sciences, poland funding plant protection research department, isfahan agricultural and natural resources research and education center (areeo), isfahan, iran. competing interests no competing interests have been declared. copyright notice © the author(s) 2018. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation nasr esfahani m. analysis of virulence and genetic variability of alternaria alternata associated with leaf spot disease in potato plants in iran. acta mycol. 2018;53(1):1105. https:// doi.org/10.5586/am.1105 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:mne2011%40gmail.com?subject=analysis%20of%20virulence%20and%20genetic%20variability%20of%20alternaria%20alternata%20associated%20with%20leaf%20spot%20disease%20in%20potato%20plants%20in%20iran https://doi.org/10.5586/am.1105 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1105 https://doi.org/10.5586/am.1105 2 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(1):1105 nasr esfahani / genetic and virulence variability of alternaria alternata adopted control measure is the use of fungicides and resistant cultivars. in iran, genetic information is available based on issr of only 11 a. alternata isolates from potato plants collected in hamedan province. moreover, no comprehensive study has analyzed the genetic diversity among the iranian a. alternata isolates from various potato-growing regions for control planning programs [10]. therefore, given the decline of resistance of potato plants to a. alternata and the lack of appropriate breeding programs, there is an urgent need to study the genetic variability within pathogen populations and should be considered one of the first steps for disease management strategy of new and aggressive isolates and evaluation of the resistance of different potato varieties [11]. thus, considering the importance of leaf spot disease in potato and a. alternata in iran [12], efforts should focus in investigating the present status of the disease. therefore, the present study aimed to estimate the variability among a. alternata isolates obtained from potato plants collected from the main potato growing regions of iran. we aimed to obtain updated and more reliable information on the virulence and genetic variability of a. alternata populations, which could aid in the selection of appropriate breeding and management strategies against the a. alternata pathogen. material and methods fungal isolates a total of 28 single-spore isolates [13] of a. alternata were obtained from the plant pathology laboratory, isfahan research and education center for agriculture and natural resources, isfahan, iran (tab. 1). the isolates were collected in 2010 from the main potato growing regions of iran, namely, ardebil, hamedan, isfahan, and fars provinces (fig. 1). population samples of a. alternata were collected in june to august 2010 from randomly selected fields of potato plants from 28 sites in four provinces. for each sampling site, leaves with a. alternata lesions were collected from the potato plants. the a. alternata populations originated from seven isolates from every province, namely, the ardebil, hamedan, isfahan, and fars provinces. conidia of all isolates were produced in chains on conidiophores and presented inverted pear shapes with short beaks and dark brown color, with dimensions of 20–50 µm × 9–18 µm and two to six (four) transversal and one to four (two) longitudinal septa. each of the 28 isolates were confirmed to be a. alternata based on the morphological characteristics [4]. virulence assay pathogenicity tests were conducted in a greenhouse using all 28 a. alternata isolates. the experiment was carried out based on a completely randomized design with 10 replicates (pots) for infected plants and control plants following the methods described by mamgain et al. [8]. potato (solanum tuberosum) ‘agria’ seed tubers were planted in plastic pots (30 cm diameter) containing soil and perlite under greenhouse conditions (18–25°c) at the plant protection research department, isfahan agricultural and natural resources research and education center, (areeo), isfahan, iran. potato tuber seeds consisted of small whole tubers weighing approximately 50 g each. one tuber seed piece was planted in each pot. tubers were treated with thiabendazole (tekto wp60%, golsam gorgan company, tehran) before planting. plants were irrigated once per day or as needed. each replication consisted of ten leaves of each plant. conidial suspension spray (103 spores/ml) was used to inoculate 1-month-old potato ‘agria’ plants after emergence. after pathogen inoculation, the plants were covered for 48 h with plastic bags in the greenhouse with 14 h of light and 10 h of darkness [14]. for estimation of the proportion of the leaf surfaces infected with a. alternata, disease rating was conducted at 7 days after inoculation based on the following 5-point scoring scale: 0 = no disease symptoms, lesions as pinpoints and nonmeasurable; 1 = <10% of the leaves have brown necrotic lesions; 2 = 10% to 25%; 3 = more than 25% to 50%; 4 = more than 50% to 75% of the leaves have brown necrotic lesions; and 5 = more than 75% to 100% of the leaves have brown necrotic lesions or appeared completely brown 3 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(1):1105 nasr esfahani / genetic and virulence variability of alternaria alternata [15]. the experiments were repeated twice, and reisolation of the inoculated fungi was performed to fulfill koch’s postulate. the following formula was used to calculate percent disease severity (pds) in each replication: pds = (∑rt × 100)/(sn), where t represents the total number of leaves in each category, r is the disease severity scale, n is the total number of leaves tested, and s is the highest number in the scale. data were transformed to arcsine square root and analyzed by conducting analysis of variance procedure (anova, p < 0.05). means were compared by performing duncan’s multiple range test using sas software version 9.2. dna extraction, pcr amplification total genomic dna was extracted from all isolates using the ctab method [16]. a nanodrop nd-1000 spectrophotometer (lms co., ltd, tokio, japan) was used to verify the quality and concentration of genomic dna extracts. to characterize the isolates, the following genes were pcr-amplified using the indicated primers: a region of the gpd tab. 1 origins and virulence variability of alternaria alternata isolates used in this study. no.1 isolate name location state percent disease severity (pds)2 1 aa-h1 asad abad hamedan 23.83 ab 2 aa-h2 shirin su hamedan 48.00 ab 3 aa-h3 kabodar ahang hamedan 49.50 ab 4 aa-h4 bahar and saleh abad hamedan 45.33 ab 5 aa-h5 hamedan hamedan 67.50 ab 6 aa-h6 qabaq tappeh hamedan 12.50 b 7 aa-h7 famenin hamedan 31.67 ab 8 aa-a1 agha bagher village ardebil 16.50 b 9 aa-a2 khalifeh lu village ardebil 28.83 ab 10 aa-a3 yunjalu village ardebil 52.33 ab 11 aa-a4 tupraghlu village ardebil 17.67 b 12 aa-a5 samian village ardebil 44.67 ab 13 aa-a6 soltan abad ardebil 42.83 ab 14 aa-a7 ardebil ardebil 37.67 ab 15 aa-f1 kushk mola village fars 48.88 ab 16 aa-f2 boroj village fars 30.67 ab 17 aa-f3 dariun village fars 36.50 ab 18 aa-f4 deh bid fars 69.17 a 19 aa-f5 shirin abad village fars 50.00 ab 20 aa-f6 hasan abad village fars 38.00 ab 21 aa-f7 bovanat village fars 36.00 ab 22 aa-i1 chadegan isfahan 46.00 ab 23 aa-i2 daran isfahan 52.50 ab 24 aa-i3 rozveh isfahan 55.00 ab 25 aa-i4 semirom isfahan 27.17 ab 26 aa-i5 golpayegan isfahan 43.33 ab 27 aa-i6 mehdi abad isfahan 38.00 ab 28 aa-i7 nisian isfahan 49.33 ab 1 alternaria alternata isolates causing leaf spot of potato. 2 values followed by the same letter in the column did not differ significantly (0.05 level) in duncan’s multiple range test. 4 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(1):1105 nasr esfahani / genetic and virulence variability of alternaria alternata gene using the primers gpd1 and gpd2 [17], alt a1 using primers alt-a1-for and alta1-rev [18], calmodulin using primers caldf1 and caldr1 [5], actin using primers actdf1 and actdr1 [5], and plasma membrane atpase using primers atpdf1 and atpdr1 [5]. pcr amplification of the gene sequences was performed as described by lawrence et al. [5]. pcr products were then purified using the gene jettm commercial pcr purification kit (fermentas, axon scientific, malaysia) and sequenced using the same forward and reverse primers by a commercial sequencing service provider (iraizol old extraction dna kits, rona bio-fanavaran co., iran). random amplified polymorphic dna (rapd) and inter simple sequence repeat (issr) analyses based on the results of initial screening against a set of representative isolates (tab. 2), we selected six out of 15 primers, namely, opa-16, opc-06, opc08, opp-16, opp-19, and opx-12 (operon technologies inc., alameda, ca, usa) for rapd analysis of the a. alternata isolates. the above-mentioned primers amplify regions with high polymorphism and reproductive profiles. for the issr analysis, nine primers, namely, ubc-807, ubc-808, ubc-809, ubc-834, ubc-835, ubc840, ubc-841, ubc-849, and ubc856, were selected among 12 primers based on the results of initial screening against a set of representative isolates (tab. 2). rapd and issr analyses were carried out as previously described by nasehi et al. [6]. fig. 1 map of iran showing the locations of potato plantations where the 28 alternaria alternata isolates were collected. the names of the isolates are based on the names indicated in tab. 1. tab. 2 rapd and issr primers utilized to identify and assess interspecific genetic diversity among alternaria alternata isolates collected from potato plants. rapd primer1 sequence (5'–3') issr primer2 sequence (5'–3') opa-03 agtcagccac ubc-807* agagagagagagagagt opa-04 aatcgggctg ubc-808* agagagagagagagagc opa-13 cagcacccac ubc-809* agagagagagagagagg opa-16* agccagcgaa ubc-818 cacacacacacacacag opb-17 agggaacgag ubc-834* agagagagagagagagyt opb-18 ccacagcagt ubc-835* agagagagagagagagyc opc-06* gaacggactc ubc-840* gagagagagagagagayt opc-08* tggaccggtg ubc-841* gagagagagagagagayc ope-01 cccaaggtcc ubc-842 gagagagagagagagayg opp-16* ccaagctgcc ubc-846 cacacacacacacacart opp-17 tgacccgcct ubc-849* gtgtgtgtgtgtgtgtya opp-18 ggcttggcct ubc-856* acacacacacacacacya opp-19* gggaaggaca opx-12* tcgccagcca opx-14 acaggtgctg 1,2 rapd and issr primers with an asterisk (*) were utilized to identify and assess interspecific genetic diversity among alternaria alternata isolates. these primers with high polymorphism and reproductive profiles were chosen among all primers based on the results of initial screening against a set of representative isolates. 5 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(1):1105 nasr esfahani / genetic and virulence variability of alternaria alternata rapd and issr analyses all pcr runs were performed in triplicate to confirm the consistency of amplification. the bands were considered as binary characters and were scored as either 1 (presence of dna band) or 0 (absence of dna band). the scores were then entered into a matrix and analyzed based on the numerical taxonomy and multivariate analysis using the program ntsys-pc 1.8 (applied biostatistics inc., setauket, ny, usa) [19]. the similarity matrix was calculated using jaccard’s similarity coefficient. clustering was performed using the unweighted pair group method using arithmetic averages (upgma) to generate the dendrogram. sequence alignment and phylogenetic analysis for molecular identification, five representative isolates, namely, aa-a5, aa-a6, aa-h2, aa-h3, and aa-i1, were selected from the 28 isolates. one isolate was selected from each of the five groups based on the rapd and issr grouping. consensus sequences were generated from the forward and reverse dna strands using bioedit sequence alignment editor [20]. to analyze the phylogenetic relationships between the a. alternata isolates and other alternaria species, the sequences of the five representative isolates from this study and sequences of all the reference of the species-groups of alternaria species [5] were aligned using the clustal w multiple alignment program [21]. stemphylium callistephi was used as the out-group taxon. we performed phylogenetic analysis of the combined dataset comprising the gpd, plasma membrane atpase, alt a1, calmodulin, and actin genes using maximum likelihood method with the kimura two-parameter model in mega 7.0 (genbank accession, fig. 2) [22]. branch support of the trees obtained from maximum likelihood analysis was assessed by bootstrapping with 1,000 replications to estimate the reliability of inferred monophyletic groups. mega7: molecular evolutionary genetics analysis version as used 7.0 for larger datasets [22]. results virulence assay symptoms of the leaf spot disease were observed in all the inoculated potato plants at 7 days after inoculation. foliar lesions were initially visible as pinpoints that were irregular to circular in shape. brown spots were usually observed first on the lower leaves and subsequently spread as circular spots throughout the leaves. control plants did not exhibit any symptoms of the disease. alternaria alternata isolates were reisolated from inoculated plants and were found to be identical to the original isolates used. these results reconfirmed the a. alternata isolates as the causal agents of leaf spot disease in the potato plants. however, we observed high variability in virulence among the four isolates (tab. 1). the aa-f4 (pds of 69.17) isolate was found to be the most virulent isolate, while the three isolates, namely, aa-h6 (pds of 12.50), aa-a1 (pds of 16.50), and aa-a4 (pds of 17.67), were the least virulent isolates. other isolates showed moderate virulence towards potato ‘agria’ plants with no significant differences in virulence. fig. 2 phylogenetic tree constructed based on the combined dataset (five target genes) using five representative isolates from this study and other alternaria species retrieved from genbank. stemphylium callistephi was used as the out-group taxon. the bar indicates the number of nucleotide substitutions per site. numbers of bootstrap support values ≥50% based on 1,000 replicates. 6 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(1):1105 nasr esfahani / genetic and virulence variability of alternaria alternata rapd analysis amplification using six rapd primers generated a total of 47 consistently amplified fragments (100 to 3,000 bp), of which 100% were polymorphic. the average number of bands per primer was 7.83. the dendrogram produced using upgma analysis based on jaccard’s coefficient indicated relatively high variability among the 28 a. alternata isolates. the jaccard’s similarity coefficient index ranged from 100% to 60%, with the mean value of 0.80 (fig. 3). the genetic diversity showed no association with the geographical origin of the isolates. issr analysis the nine issr primers generated a total of 61 consistently amplified dna bands, of which 83.61% were polymorphic. the average number of bands per primer was 6.8. the bands ranged in size from approximately 100 to 3,000 bp. the dendrogram also indicated high variability among the 28 a. alternata isolates and grouped the isolates into different groups, with no correlation with the geographical regions of the isolates (fig. 4). the jaccard’s similarity coefficient ranged from 100% to 68% with the mean value of 0.84. phylogenetic analysis pcr amplification of the genes corresponding to gpd, plasma membrane atpase, alt a1, calmodulin, and actin in the five representative isolates produced fragments with sizes 797, 1,194, 467, 780, and 939 bp, respectively. phylogenetic analysis of the combined dataset indicated that the five representative isolates were clustered in a subdistinct cluster, which included a. alternata with a high bootstrap value of 94% (fig. 2). the five isolates were clearly distinguished from other selected alternaria species, which were found to be highly similar to the a. alternata isolates in the present study. stemphylium callistephi was phylogenetically distant to alternaria and clustered as its sister taxon. discussion results of the virulence assays indicated that all the a. alternata isolates were pathogenic against potato ‘agria’ plants, and disease symptoms were observed in the leaves of all potato ‘agria’ plants inoculated with a. alternata. reisolation of a. alternata from all the inoculated samples confirmed koch’s postulates. therefore, all a. alternata isolates were verified to be the causal agents of the leaf spot disease in the potato ‘agria’ plants. fig. 3 dendrogram generated via the upgma method showing the genetic variability of the 28 alternaria alternata isolates based on six rapd primers (opa-16, opc-06, opc-08, opp-16, opp-19, and opx-12). the names of the isolates are based on the names indicated in tab. 1. fig. 4 dendrogram generated via the upgma method showing the genetic variability of 28 alternaria alternata isolates derived from a combination of nine issr primers (ubc-807, ubc-808, ubc-809, ubc-834, ubc-835, ubc-840, ubc-841, ubc-849, and ubc-856). the names of the isolates are based on the names indicated in tab. 1. 7 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(1):1105 nasr esfahani / genetic and virulence variability of alternaria alternata our findings also revealed high variability in virulence among the a. alternata isolates. in addition, the grouping of the isolates based on virulence was not found to be correlated with the results of rapd and issr analyses and the geographical origins of the isolates. therefore, further detailed studies must be conducted using a larger number of a. alternata isolates. our current findings were consistent with those of previous studies on a. alternata isolates obtained from citrus hybrids [23], potato plants [10,24], and tomato plants [24,25] throughout iran. the rapd and issr techniques are extremely powerful tools for distinguishing among individuals that exhibit intraspecific and interspecific variability [26]. in the present study, 100% and 84% of the isolates were found to be polymorphic with respect to the rapd and issr markers, respectively. the similarity indices were 84% and 80% for the rapd and issr markers, respectively. rapd and issr markers exhibit lower polymorphism rates than those of aflp and other molecular markers. however, the aflp method is more laborious, time-consuming and costly and involves the use of radioactive reagents. therefore, the rapd and issr techniques could serve as reliable alternatives for molecular characterization because they are easier to implement and are less costly [9,24]. the rapd and issr methods have been employed to generate comprehensive information regarding the genetic variability of a. alternata populations in citrus [23], linseed [27], pistachio [28], potato [10,24], tangerine [13], and tomato [24,25]. these methods have also been used to investigate the genetic variability of other fungal species, such as stemphylium lycopersici, which is associated with leaf spot disease in vegetable crops [26]. in the present study, the results of rapd and issr analyses were very similar and showed that the markers were 84% to 100% polymorphic, respectively. therefore, the 28 a. alternata isolates exhibited relatively high diversity in the main potato-growing regions of iran. the rapd and issr markers showed no correlation with the geographical origins of the isolates, suggesting that the isolates are widely dispersed across iran. the above results were consistent with those of a previous study, in which issr markers were used to assess the genetic diversity of 11 a. alternata isolates that caused brown leaf spot disease in potato plants throughout iran [10]. furthermore, our findings indicated that both rapd and issr markers can be used to reliably evaluate the diversity of a. alternata isolates. recently, sequencing of different dna regions has been used as an alternative to morphology-based identification of alternaria species [5,7,29] and other fungal species. consequently, different dna regions have been used for the characterization and identification of various alternaria species [5]. studies have indicated that the combination of various dna genes, including gpd, plasma membrane atpase, alt a1, calmodulin, and actin, can successfully determine the monophyly of alternaria species. the above-mentioned genes showed higher polymorphism than those of the translation elongation factor 1-alpha (tef) and beta-tubulin genes. in the present study, phylogenetic analysis of the combined dataset comprising the gpd, plasma membrane atpase, alt a1, calmodulin, and actin genes confirmed that all five representative isolates, namely, aa-a5, aa-a6, aa-h2, aa-h3, and aa-i1, were a. alternata species. furthermore, we detected no significant differences in the sequences of the five target genes among the representative isolates. conclusions in conclusion, rapd and issr markers were successfully used to genetically distinguish all the a. alternata isolates. the results of the present study revealed the extant diversity in pathogenicity and genetics of a. alternata isolates. therefore, new and aggressive isolates should be screened to evaluate the potential resistance of potato varieties, which would be useful for the selection of appropriate breeding strategies against the potato pathogen a. alternata. to our knowledge, no comprehensive study has investigated the genetic diversity of a. alternata isolates that cause leaf spot disease in potato plants from various geographical regions in iran. rapd and issr markers could also be used to investigate a broader range of a. alternata isolates from other regions of the world and can also be extended to other host plants. given that temperature, wind speed, potato cultivar, and other factors can affect the population structure of fungi, 8 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(1):1105 nasr esfahani / genetic and virulence variability of alternaria alternata acknowledgments we thank the plant protection research division, isfahan center for agricultural and natural resources research and education, isfahan, iran and also, plant protection research institute, tehran, iran, for providing facilities to implement the project. references 1. andersen b, hansen me, smedsgaard j. automated and unbiased image analyses as tools in phenotypic classification of small-spored alternaria spp. phytopathology. 2005;95:1021–1029. https://doi.org/10.1094/phyto-95-1021 2. droby s, dinoor a, prusky d, barkai-golan r. pathogenicity of alternaria alternata on potato in israel. phytopathology. 1984;74:537–542. https://doi.org/10.1094/phyto-74-537 3. rotem j. the genus alternaria; biology, epidemiology, and pathogenicity. saint paul, mn: american phytopathological society press; 1998. 4. van der waals je, pitsi be, marais c, wairuri ck. first report of alternaria alternata causing leaf blight of potatoes in south africa. plant dis. 2011;95(3):363–363. https://doi.org/10.1094/pdis-11-10-0820 5. lawrence dp, gannibal pb, peever tl, pryor bm. 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alternaria alternata isolates causing potato brown leaf spot, using issr markers in iran. j plant pathol microbiol. 2015;6:286. https://doi.org/10.4172/2157-7471.1000286 11. mcdonald ba, linde c. pathogen population genetics, evolutionary potential, and durable resistance. annu rev phytopathol. 2002;40:349–379. https://doi.org/10.1146/annurev.phyto.40.120501.101443 12. nasr esfahani m. identification of ulocladium atrum causing potato leaf blight in iran. phytopathol mediterr. 2018;57(1):112–114. https://doi.org/10.14601/phytopathol_mediterr-22282 13. demartelaere acf, nascimento lc, marinho co, nunes mc, abraão pc, gomes rss, et al. diversity among isolates of the tangerine pathotype of alternaria alternata. american journal of plant sciences. 2018;9:81987. https://doi.org/10.4236/ajps.2018.92017 14. shahbazi h, aminian h, sahebani n. halterman d. effect of alternaria solani exudates on resistant and susceptible potato cultivars from two different pathogen isolates. plant pathol j. 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(practical approach series). 17. berbee ml, pirseyedi m, hubbard s. cochliobolus phylogenetics and the origin future studies should investigate the population structure of a. alternata under various growing conditions. in addition, further studies are required to determine pathogen specialization of different cultivars. https://doi.org/10.1094/phyto-95-1021 https://doi.org/10.1094/phyto-74-537 https://doi.org/10.1094/pdis-11-10-0820 https://doi.org/10.3852/12-249 https://doi.org/10.1007/s11557-014-0991-1 https://doi.org/10.3114/sim0015 https://doi.org/10.1111/ppa.12734 https://doi.org/10.4172/2157-7471.1000286 https://doi.org/10.1146/annurev.phyto.40.120501.101443 https://doi.org/10.14601/phytopathol_mediterr-22282 https://doi.org/10.4236/ajps.2018.92017 https://doi.org/10.5423/ppj.2011.27.1.014 9 of 9© the author(s) 2018 published by polish botanical society acta mycol 53(1):1105 nasr esfahani / genetic and virulence variability of alternaria alternata of known, highly virulent pathogens, inferred from its and glyceraldehyde3-phosphate dehydrogenase gene sequences. mycologia. 1999;91:964–977. https://doi.org/10.2307/3761627 18. hong sg, cramerb ra, lawrence cb, pryora bm. alt a 1 allergen homologs from alternaria and related taxa: analysis of phylogenetic content and secondary structure. fungal genet biol. 2005;42:119–129. https://doi.org/10.1016/j.fgb.2004.10.009 19. rohlf ej. ntsys-pc: numerical taxonomy and multivariate analysis system, version 1.80. setauket, ny: applied biostatistics, inc.; 1993. 20. hall ta. bioedit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/nt. nucleic acids symp ser (oxf ). 1999;41:95–98. 21. thompson jd, higgins dg, gibson tj. clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice. nucleic acids res. 1994;22:4673–4680. https://doi.org/10.1093/nar/22.22.4673 22. kumar s, stecher g, tamura k. mega7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. mol biol evol. 2016;33(7):1870–1874. https://doi.org/10.1093/molbev/msw054 23. kakvan n, zamanizadeh h, morid b, taheri h, hajmansor s. study on pathogenic and genetic diversity of alternaria alternata isolated from citrus hybrids of iran, based on rapd-pcr technique. eur j exp biol. 2012;2(3):570–576. 24. weir tl, huff dr, christ bj, romaine cp. rapd-pcr analysis of genetic variation among isolates of alternaria solani and alternaria alternata from potato and tomato. mycologia. 1998;90(5):813–821. https://doi.org/10.2307/3761323 25. morris pf, connolly ms, st clair da. genetic diversity of alternaria alternata isolated from tomato in california assessed using rapds. mycol res. 2000;104(03):286–292. https://doi.org/10.1017/s0953756299008758 26. nasehi a, kadir jb, nasr esfahani m, abed ashtiani f, wong my, rambe sk, et al. analysis of genetic and virulence variability of stemphylium lycopersici associated with leaf spot of vegetable crops. eur j plant pathol. 2014;140(2):261–273. https://doi.org/10.1007/s10658-014-0460-3 27. kale sm, pardeshi vc, gurjar gs, gupta vs, gohokar rt, ghorpade pb, et al. intersimple sequence repeat markers reveal high genetic diversity among isolates of indian origin. j mycol plant pathol. 2012;42(2):194. 28. pryor bm, michailides tj. morphological, pathogenic, and molecular characterization of alternaria isolates associated with alternaria late blight of pistachio. phytopathology. 2002;92(4):406–416. https://doi.org/10.1094/phyto.2002.92.4.406 29. woudenberg jhc, seidl mf, groenewald jz, de vries m, stielow jb, thomma bphj, et al. alternaria section alternaria: species, formae speciales or pathotypes? stud mycol. 2015;82:1–21. https://doi.org/10.1016/j.simyco.2015.07.001 https://doi.org/10.2307/3761627 https://doi.org/10.1016/j.fgb.2004.10.009 https://doi.org/10.1093/nar/22.22.4673 https://doi.org/10.1093/molbev/msw054 https://doi.org/10.2307/3761323 https://doi.org/10.1017/s0953756299008758 https://doi.org/10.1007/s10658-014-0460-3 https://doi.org/10.1094/phyto.2002.92.4.406 https://doi.org/10.1016/j.simyco.2015.07.001 abstract introduction material and methods fungal isolates virulence assay dna extraction, pcr amplification random amplified polymorphic dna (rapd) and inter simple sequence repeat (issr) analyses rapd and issr analyses sequence alignment and phylogenetic analysis results virulence assay rapd analysis issr analysis phylogenetic analysis discussion conclusions acknowledgments references 2018-06-29t22:42:17+0100 piotr otręba professor dr hab. anna maria bujakiewicz 1 of 3published by polish botanical society acta mycologica scientists professor dr hab. anna maria bujakiewicz anna kujawa institute for agricultural and forest environment of polish academy of sciences, research station in turew, turew, szkolna 4, 64-000 kościan, poland * email: anna.kujawa@isrl.poznan.pl abstract the article presents the biography and scientific achievements of professor anna bujakiewicz. after receiving her master’s degree and doctorate in biology and mycology from adam mickiewicz university in poznań, professor bujakiewicz continued her exciting research and teaching on mycology at her alma mater posnaniensis for more than 50 years. her publications in this field include many books, articles, and other scholarly reports. keywords mycology; mycocoenology; macromycetes; poznań this issue of acta mycologica is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. professor anna maria bujakiewicz, a native-born wielkopolska province inhabitant, has always been interested in nature. following her passion, in 1957 she started studies at the institute of biology at adam mickiewicz university in poznań. she described her fascination as follows: “i chose biology because my childhood and my youth passed amid my beloved nature. i especially loved botany and ornithology. what fascinated me the most was observing nature during field classes which took place in different parts of poland” [1]. she graduated in 1962 after defending her thesis, entitled “udział grzybów wyższych w grądach okolic opalenicy (uroczysko urbanowo i dakowy mokre, nadleśnictwo podłoziny) [macromycetes in oak-hornbeam forests near opalenica (uroczysko urbanowo i dakowy mokre, forest district podłoziny)]”, supervised by prof. dr. hab. zygmunt czubiński, and was employed at the department of plant systematics and geography at adam mickiewicz university. she looks back to her studies and the beginning of her work with pleasure, and she also recalls her professors, zygmunt czubiński, teofil wojterski, stefan krupko, jan rafalski, anna czekalska, jan czekanowski, izabela dąmbska, and stanisław lisowski. forests, especially alluvial and alder forests, fascinated her, and she devoted most of her time to studying them. her focus was on mutual relationships between plants and fungi. her first mycocoenological studies were planned according to the directions of the pioneer of polish mycocoenology, professor andrzej nespiak. the results of the mycocoenological studies conducted in riparian and alder forests doi: 10.5586/am.1060 publication history received: 2015-06-27 accepted: 2015-07-23 published: 2015-08-05 handling editor maria rudawska, institute of dendrology of the polish academy of sciences, poland funding this work did not involve any funding. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation kujawa a. professor dr hab. anna maria bujakiewicz. acta mycol. 2015;50(1):1060. http:// dx.doi.org/10.5586/am.1060 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:anna.kujawa%40isrl.poznan.pl?subject=professor%20dr%20hab.%20anna%20maria%20bujakiewicz http://dx.doi.org/10.5586/am.1060 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1060 http://dx.doi.org/10.5586/am.1060 2 of 3© the author(s) 2015 published by polish botanical society acta mycol 50(1):1060 kujawa / professor dr hab. anna maria bujakiewicz became the basis for her doctoral thesis, entitled “udział grzybów wyższych w olesach i lasach łęgowych wielkopolski [macromycetes in the alder and riparian forests of wielkopolska province]”, promoted by prof. dr. hab. teofil wojterski. mycocoenology was not the only passion of professor bujakiewicz. over the next years, she combined her love for mountain hiking with attempts to observe regularities in fungi occurrence in forest associations in uphill gradients. several years of analyzing mt babia góra and strenuous mycocoenological research provided the basis for her postdoctoral thesis, entitled “grzyby babiej góry ii. wartość wskaźnikowa macromycetes w zespołach leśnych [fungi of mt babia góra ii. the indicating value of macromycetes in forest associations]”, defended in 1979. she also gained experience abroad on scholarships in the netherlands, sweden, and norway. in the years 1982–1983 she studied in the usa, at cornell university, ithaca, new york, under the direction of a famous worldwide fungi taxonomer, professor richard p. korf. she received her nomination for professorship in 1994. professor anna bujakiewicz’s research has focused on mycocoenological issues until today. she has analyzed the macromycetes in forests both in poland and abroad (usa, the netherlands, sweden, norway, and finland). in poland, she analyzed forest associations in the beech forest near szczecin and the babia góra, słowiński, wielkopolski, and białowieża national parks. she conducted mycological research in riparian and alder forests of many nature reserves and took part in the crypto project in białowieża forest, conducted by professor janusz b. faliński. she also organized several mycological and botanical expeditions for foreigners, among which were expeditions for the international society for vegetation sciences (1978) and an expedition during the international botanical congress in western berlin (1987). since the beginning of her work at adam mickiewicz university, professor anna bujakiewicz has dreamed of implementing mycology as a separate subject in the program in the faculty of biology. she managed to obtain her goal in the academic year 1998–1999. earlier, in 1991, she had established a mycological section of the polish botanical society in poznań, where she has resided until today and with whom she has conducted over 200 seminars with lecturers from all over poland. professor anna bujakiewicz has been able to instill her passion among a great number of students. the latest experiment is a lecture, “biology, ecology, and fungi protection”, in english for students of environment protection studies within the human resources of the eu program. bujakiewicz’s fascination with fungi necessitated that she popularize the study of these organisms, and she therefore conducted expeditions and corresponded with amateurs. each september from 1996 to 2005, she visited białowieża forest and organized fungi exhibitions visited by tourists, białowieża residents, and students in the nature education centre at białowieża national park. since 2005, the organization of this exhibition has been taken over by a new generation of researchers who have cooperated with her in previous years. during her work, professor anna bujakiewicz has participated actively in mycological congresses, conferences, and symposiums in poland and abroad. her scientific experience includes 133 polish and foreign publications, among others the summary of macrofungi in deciduous forests [2], remarks on macrofungi occurring in boreal and temperate grey alder [3], and macrofungi in the alnetum incanae association in the western carpathians [4]. ecological research on fungi occurrence patterns in black alder bog forests were described in an article from the olszyny niezgodzkie nature reserve [5]. she is also a co-author of an elaboration on mycocoenological studies covering half a century, 1952–2002 [6]. a number of people gained mycological experience under the supervision of professor bujakiewicz. these include 14 bachelors, 21 masters, and 3 doctors (anna kujawa, marek halama, and mateusz stefaniak). professor anna bujakiewicz has reviewed five doctoral dissertations, three applications for professorship, and an application by the wrocław university senate to award a degree of honoris causa of wrocław university to mark r.d. seaward, a professor at bradford university in england. her experience and professionalism have been appreciated in the professional and organizational field. in the years 1984–1985, professor bujakiewicz held the position of deputy dean of the department of biology at adam mickiewicz university, and in 1992–2005 she was the head of the department of plant ecology and environment 3 of 3© the author(s) 2015 published by polish botanical society acta mycol 50(1):1060 kujawa / professor dr hab. anna maria bujakiewicz protection. she has taken an active role in activities of scientific societies, and she remains a well-known expert in the field. during 2004–2009, she was a member of the botanical committee of the polish academy of sciences, and in 2005–2007 a member of the ecology committee of the polish academy of sciences. since 2004, she has been a member of the department of mathematics and nature at the poznań society of friends of learning. in the years 2001–2014, she was a member of the zygmunt czubiński medal chapter. through the years, she has participated in scientific sessions of the babia góra and białowieża national parks. she is a member of the polish botanical society and the polish mycological society. since 1980, she has performed international activities as a member of the iavs (international association for vegetation science). she was awarded the second stage minister’s prize for scientific studies and was honored for studies in the białowieża national park. in 2010, she received a medal of professor bolesław hryniewiecki to appreciate her development and popularization of botanical knowledge. her work she summarizes as follows: “in my opinion, i chose the direction of my studies correctly and i fulfilled my professional aims. i have always treated my work as a duty. i was taught this attitude by my father”[1]. references 1. bujakiewicz a, dunajska-zielińska l, kopczyńska-idzikowska k, malinowski a. 50 lat my z biologią – 50 lat biologia z nami. 1962–2012. poznań: wydział biologii uam, wydawnictwo kontekst; 2012. 2. bujakiewicz a. macrofungi on soil in deciduous forests. in: winterhoff w, editor. fungi in vegetation science. dordrecht: kluwer academic publishers. 1992. p. 49–78. (handbook of vegetation science; vol 19). http://dx.doi.org/10.1007/978-94-011-2414-0 3. bujakiewicz a. general remarks on macrofungi occurring in boreal and temperate grey older forests. blyttia. 1993;51(3–4):99–110. 4. bujakiewicz a. macrofungi in the alnetum incanae association along jaworzyna and skawica river valleys (western carpathians) pol bot j. 2011;56(2):267–285. 5. bujakiewicz a. response of macrofungi to mosaic arrangement of biotic microforms in the ribo nigri-alnetum in the olszyny niezgodzkie reserve. acta mycol. 1999;34(2):267– 280. http://dx.doi.org/10.5586/am.1999.016 6. ławrynowicz m, bujakiewicz a, mułenko w. mycocoenological studies in poland – 1952– 2002. łódź: polish botanical society; 2004. (monographiae botanicae; vol 93). http:// dx.doi.org/10.5586/mb.2004.001 http://dx.doi.org/10.1007/978-94-011-2414-0 http://dx.doi.org/10.5586/am.1999.016 http://dx.doi.org/10.5586/mb.2004.001 http://dx.doi.org/10.5586/mb.2004.001 abstract references 2015-08-02t15:22:02+0100 piotr otręba preliminary studies of fungi in the biebrza national park (ne poland). part iii. micromycetes – new data 1 of 28published by polish botanical society acta mycologica original research paper preliminary studies of fungi in the biebrza national park (ne poland). part iii. micromycetes – new data malgorzata ruszkiewicz-michalska1*, stanisław bałazy2, jerzy chełkowski3, maria dynowska4, julia pawłowska5, ewa sucharzewska4, jarosław szkodzik6, cezary tkaczuk7, mateusz wilk8,9, marta wrzosek5 1 department of algology and mycology, university of łódź, banacha 12/16, 90-237 łódź, poland 2 institute of agricultural and forest environment, polish academy of sciences, bukowska 19, 60-809 poznań, poland 3 institute of plant genetics, polish academy of sciences, strzeszyńska 34, 60-479 poznań, poland 4 department of mycology, university of warmia and mazury in olsztyn, oczapowskiego 1a, 10-957 olsztyn, poland 5 department of molecular phylogenetics and evolution, university of warsaw, al. ujazdowskie 4, 00-478 warsaw, poland 6 nature & ecology of łódź macroregion website, ekolodzkie.pl 7 department of plant protection and breeding, siedlce university of natural sciences and humanities, prusa 14, 08-110 siedlce, poland 8 department of plant ecology and environmental protection, university of warsaw, al. ujazdowskie 4, 00-478 warsaw, poland 9 college of inter-faculty individual studies in mathematics and natural sciences, university of warsaw, żwirki i wigury 93, 00-089 warsaw, poland * corresponding author. email: mrusz@biol.uni.lodz.pl abstract ecological information concerning 292 fungal taxa is reported as a result of two surverys in the biebrza national park. most data presented come from the 5-day all-fungi inventory of the polish mycological society in 2013, and 47 species were recorded during studies in the biele suchowolskie fen in 2008/2009. in total, 27 species of zygomycetes, 232 ascomycetes (including anamorphs) and 27 basidiomycetes (mainly pucciniales). additionaly some representatives of fungi-like organisms from stramenopiles (4 species) and dictyostelia (2) were identified. fungal groups included were the same as in the previous survey in 2012: 190 taxa associated with plants, 15 with animals, 8 with fungi and 71 isolated from soil, plant debris and animal excrements. the most numerous were anamorphic ascomycetes (159 species). nineteen species have not been previously known from poland and 31 species are rare (1–3 localities). for the biebrza national park 197 species (67.5%) are new. keywords all-species inventory; micromycetes; plant parasites; arthropod pathogens; soil fungi; fungal ecology; protected area this paper is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. introduction the all-fungi inventory carried out by the polish mycological society (pms) was organized in the biebrza national park (bbnp) in 2012 and 2013. the results of the first year of study as well as general state of knowledge about bbnp natural environment were presented in 2012 [1,2]. although the studies in 2013 were continuation of the doi: 10.5586/am.1067 publication history received: 2015-11-03 accepted: 2016-01-08 published: 2016-01-29 handling editor tomasz leski, institute of dendrology of the polish academy of sciences, poland authors’ contributions all authors: collecting and identification of the materials; mrm, mwr, jp, ct, js: writing the manuscript; mrm, mwr, jp: designing the study; mwi: text proofreading and identification of selected materials funding the works in biele suchowolskie fen in 2008/2009 were carried out as a part of a grant from the polish ministry of science and higher education (no. nn305433933). the study was also partially supported by the university of łódź (grant no. 506/1144 in 2015). funding from eu through the european social fund, contract number udapokl.04.01.01-00-072/09-00 to mwi is acknowledged. competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation ruszkiewicz-michalska m, bałazy s, chełkowski j, dynowska m, pawłowska j, sucharzewska e, et al. preliminary studies of fungi in the biebrza national park (ne poland). part iii. micromycetes – new data. acta mycol. 2015;50(2):1067. http://dx.doi. org/10.5586/am.1067 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:mrusz%40biol.uni.lodz.pl?subject=preliminary%20studies%20of%20fungi%20in%20the%20biebrza%20national%20park%20%28ne%20poland%29.%20part%20iii.%20micromycetes%20%e2%80%93%20new%20data http://dx.doi.org/10.5586/am.1067 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1067 http://dx.doi.org/10.5586/am.1067 2 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park first inventory, they were focused on new, previously unexplored areas. the aim of this paper is to present new data on micromycetes associated with plants, fungi and animals. most of them were collected in 2013 during the second survey of pms in the bbnp. some additional materials from 2012, previously unpublished, are also included here. these data are supplemented with unpublished results of the earlier study (2008/2009) devoted to soiland litter-inhabiting micromycetes of burnt and unburnt areas of the biele suchowolskie fen [3]. a synopsis of macromycetes collected during the same survey in 2013 is given in the paper by kujawa and co-workers in this issue [4]. material and methods the materials were collected on 26–31 aug. 2013 during all-fungi inventory of the pms in 5 protective units (brzeziny, grzędy, kapice, osowiec, werykle) and on 20 jun. 2008 and 15 oct. 2009 at the biele suchowolskie fen (kopytkowo protective unit). six localities (fig. 1) and seven habitats were screened: caricion nigrae alliance, continental swamp/bog pine forest (vaccinio uliginosi-pinetum), filipendulion alliance with deschampsia cespitosa (l.) beauv., middle-european alder fen forest (carici elongatae-alnetum), moderately moist, wet meadow with festuca rubra l., salicetum pentandro-cinereae in association with urtica dioica l., subatlantic swamp birch forest (vaccinio uliginosi-betuletum pubescentis), subcontinental lime-oak-hornbeam forest (tilio-carpinetum). the characteristics of particular sites including plant community fig. 1 localization of observation sites in the biebrza national park ([2], modified). 3 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park information (if available) is given in detailed part of the “results” chapter. fungi were also collected in deciduous and mixed forests (without determination of their syntaxonomic affinity) and from other habitats, including forest edges, causeways, roadsides, pathways, meadows, etc. more detailed characteristics of sites from kopytkowo protected unit were provided by several authors [3,5–7]. fungi were collected and identified according to taxon-specific methods [8–26]; for details see first part of the inventory in bbnp [1]. host plant species were determined using the key by rutkowski [27]. taxonomic system of fungi and fungus-like organisms of adl et al. [28] and hibbett et al. [29] is adopted, while nomenclature follows keller [30], humber [31], mułenko et al. [32], and index fungorum [33]. names of plants and arthropods are given after mirek et al. [34] and schaefer [35], respectively, while syntaxonomic nomenclature follows matuszkiewicz [36]. the specimens documenting research are deposited in the herbarium of the faculty of biology, university of warsaw (wa), in the fungal collection of the herbarium universitatis lodziensis (lod), as well as in fungal collection of department of plant protection and breeding of the siedlce university of natural sciences and humanities, and in the fungal collection in department of mycology of the warmia and mazury university in olsztyn. the following abbreviations and symbols are used in the list of species: bpu – brzeziny protective unit; gpu – grzędy protective unit; kpu – kapice protective unit; kopu – kopytkowo protective unit; opu – osowiec protective unit; wpu – werykle protective unit; ce-a – carici elongatae-alnetum; fdc – filipendulion alliance with deschampsia cespitosa; m-a – wet meadow with festuca rubra; s-cn – caricion nigrae; sp-c – salicetum pentandro-cinereae in association with urtica dioica; t-c – tiliocarpinetum; vu-b – vaccinio uliginosi-betuletum pubescentis; vu-p – vaccinio uliginosi-pinetum; (a) – anamorphic stage; (t) – teleomorphic stage; (0, i, ii, iii) – stages of rust fungi; – taxon new to poland; – taxon new to bbnp; # – species very rare in poland (1–3 localities). initials of the names are given to denote collecting and/or identifying person, namely: ab – abiba boulahdjel, ct – cezary tkaczuk, es – ewa sucharzewska, jc – jerzy chełkowski, js – jarosław szkodzik, md – maria dynowska, mrm – małgorzata ruszkiewicz-michalska, mw – marta wrzosek, mwk – mateusz wilk, sb – stanisław bałazy. results list of 292 species of fungi and fungi-like organisms reported here consists of 245 species collected during 5-day survey of pms, and 47 species obtained from studies at the biele suchowolskie fen in 2008–2009. the predominant group were ascomycetes (232 species), recorded mainly in anamorphic state (159 species). zygomycetes and basidiomycetes were less numerous – 27 species each (tab. 1, tab. 2). fungi-like organisms were represented by four stramenopilean species (peronosporales) and two dictyosteloid species. the most numerous group were plant-associated fungi and stramenopilean fungi-like organisms (190 taxa), followed by 15 species of arthropod (insects, spiders) pathogens and 8 species inhabiting other fungi. second numerous group (71 species) constitute fungi and dictyostelia isolated from soil and remnants of plants and animal excrements. nineteen species reported here were found in poland for the first time and about 67% (197 species) of the fungi recorded are new for the biebrza national park. thirty-one species are rarely recorded in poland – in that group only taxa with 1–3 known localities are classified. plant-associated fungi among 190 taxa found in association with plants predominate ascomycetous anamorphs (105 species, 55%), with capnodiales (59 species), pleosporales (14) and xylariales (11) as the most numerous groups. basidiomycetes and fungi-like species were represented mainly by pucciniales (25) and peronosporales (4), respectively. ascomycetes and their anamorphs were recorded mostly from leaves and only 19 species 4 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park inhabiting dead wood (branches, logs, stumps) of deciduous trees and scots pines were collected. they belonged predominantly to xylariales as well as to chaetosphaeriales, coronophorales, diaporthales, hypocreales and sordariales. fourteen species are reported from poland for the first time, including daldinia decipiens, five discosia species, mycosphaerella osborniae, nemania aenea, phaeosphaeria inclusa, phomopsis amygdaliana, seimatosporium foliicola and sphaceloma viburni which have been very rarely reported worldwide [20,37]. twenty-five species, mainy ascomycetous anamorphs, are known in poland from 1–3 localities [32,38]. among them three species have been sporadically reported in poland: helminthosphaeria clavariarum (on clavulina sp.) recorded for the first time in 2013 [39], as well as hypomyces rosellus (on deciduous wood) and stephanoma strigosum (on humaria hemisphaerica), with the last localities known from 1908 and 1936, respectively [32,38]. alternaria alternata (fr.) keissl. s. l., (a), on alliaria petiolata (bieb.) cavara & grande, opu, góra skobla, deciduous forest edge, 27 aug. 2013, leg. es, det. md & es. alternaria sp., (a), on filipendula ulmaria (l.) maxim, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; on plantago lanceolata l., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. md & es. ampelomyces quisqualis ces. s. l., (a), on erysiphe alphitoides (on quercus robur), gpu, vu-p, 29 aug. 2012, leg. & det. mrm; on erysiphe cynoglossi (on pulmonaria officinalis), bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm; on erysiphe friesii (on rhamnus cathartica), opu, sośnia, mixed forest near dune, 28 aug. 2013, leg. mw, det. mrm; on erysiphe heraclei (on heracleum sphondylium), bpu, grobla honczarowska causeway, 26 aug. 2013, leg. & det. mrm; (on peucedanum cervaria), gpu, deciduous thicket, small glade, 27 aug. 2013, leg. & tab. 1 numbers of species of fungi and fungus-like organisms collected at the study area. ta xo n an am or ph ic fu ng i c ap no di al es c ha et os ph ae ri al es c or on op ho ra le s d ia po rt ha le s d ic ty os te lia d ot hi de al es en to m op ht ho ra le s er ys ip ha le s ex ob as id ia le s h el ot ia le s h yp oc re al es m or tie re lla le s m uc or al es n eo zy gi ta le s pe ro no sp or al es pl eo sp or al es pu cc in ia le s r hy tis m at al es so rd ar ia le s v en tu ri al es x yl ar ia le s z oo pa ga le s in to ta l n o. 15 9 2 1 1 1 2 1 8 30 2 1 5 1 12 2 4 4 25 1 3 1 22 4 29 2 species recorded in anamorphic state, with known or unknown/uncertain anamorph–teleomorph connections. tab. 2 affinity of fungi taxa (according to [33]) collected/identified in anamorphic state. ta xo n b ot ry os ph ae ri al es c ap no di al es d ia po rt ha le s eu ro tia le s h el ot ia le s h yp oc re al es in ce rt ae s ed is , d ot hi de om yc et es in ce rt ae s ed is , pe zi zo m yc ot in a in ce rt ae s ed is , so rd ar io m yc et es m ic ro as ca le s pl eo sp or al es r hy tis m at al es so rd ar ia le s x yl ar ia le s in to ta l n o. 3 59 7 13 4 31 5 3 6 3 13 1 1 10 15 9 5 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park det. mrm; on erysiphe sp. (on syringa vulgaris), opu, sośnia, village, near a house, 28 aug. 2013, leg. mw, det. mrm; on golovinomyces sordidus, (on plantago major), opu, góra skobla, deciduous thicket, 27 aug. 2013, leg. md, det. md & es; on podosphaera fusca (on melampyrum nemorosum), bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. annulohypoxylon multiforme (fr.) y.m. ju, j.d. rogers & h.m. hsieh, (t), on betula pendula roth log, gpu, deciduous thicket, 27 aug. 2013, leg. & det. js. apiognomonia errabunda (roberge ex desm.) höhn. [= discula umbrinella (berk. & broome) morelet], (a), on tilia cordata mill., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. mrm. #ascochyta equiseti (desm.) grove, (a), on equisetum palustre l., bpu, długa luka overpass, sedge patch, 26 aug. 2013, leg. & det. mrm. asteroma frondicola (fr.) m. morelet, (a), on populus nigra l., bpu, grobla honczarowska causeway, 26 aug. 2013, leg. & det. mrm; on populus tremula l., gpu, deciduous thicket, 27 aug. 2013, leg. & det. mrm. #asteroma padi dc., (a), on cerasus avium (l.) moench, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es, rev. mrm; same locality and date, leg. & det. mrm; opu, carska droga, catwalk, 26 aug. 2013, leg. & det. mrm. asteromella ludwigii petr., (a), on chamaenerion angustifolium (l.) scop., wpu, carska droga near grobla honczarowska causeway, roadside, 26 aug. 2013, leg. & det. mrm. #asteromella pruni-mahaleb (pass.) bedlan [= phyllosticta passerinii berl. & voglino], (a), on cerasus avium (l.) moench, opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es, rev. mrm. asteromella sp., (a), on alliaria petiolata (m. bieb.) cavara & grande, kpu, t-c, 28 aug. 2012, leg. mrm, det. mwk & mrm; on filipendula ulmaria (l.) maxim, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm; on peucedanum oreoselinum (l.) moench, gpu, edge of dune, 30 aug. 2012, leg. mrm, det. mrm & mwk; on peucedanum cervaria (l.) lapeyr., gpu, deciduous thicket, small glade, 27 aug. 2013, leg. & det. mrm. notes. no asteromella species were described on alliaria, filipendula and peucedanum species [40]. asteromella tiliicola (oudem.) arx, (a), on tilia cordata mill., opu, biały grąd, deciduous forest, 27 aug. 2013, leg. es, det. mrm. asteromella trautmanniana (moesz) moesz, (a), on sorbus aucuparia l. emend. hedl., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm; gpu, deciduous thicket, 27 aug. 2013, leg. & det. mrm; vu-p, 29 aug. 2012, leg. & det. mrm; opu, sośnia, dune, 27 aug. 2013, leg. & det. mrm. bertia moriformis (tode) de not., (t), on pinus sylvestris l. wood, gpu, mixed forest, 28 aug. 2013, leg. & det. js. botrytis cinerea pers., (a), on potentilla argentea l. (det. j. kołodziejek), kpu, t-c, glade, 28 aug. 2012, leg. & det. mrm. cercospora armoraciae sacc., (a), on berteroa incana (l.) dc., gpu, dune, 29 aug. 2013, leg. & det. mrm. 6 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park notes. notes. the fungus already reported from bbnp as cercospora berteroae hollós [1], recently synonymized with c. armoraciae [41]. cercospora mercurialis pass., (a), on mercurialis perennis l., gpu, mixed forest, 29 aug. 2012, leg. mrm, det. mwk; deciduous forest, 27 aug. 2013, leg. & det. mrm. cercospora zebrina pass., (a), on trifolium alpestre l., gpu, edge of dune and deciduous forest, 27 aug.2013, leg. & det. mrm; on trifolium arvense l., gpu, dune, 27 aug. 2013, leg. & det. mrm. cercosporella lindaviana (jaap) u. braun, (a), on vicia cracca l., gpu, dune, 30 aug. 2012, leg. & det. mrm. cercosporella virgaureae (thüm.) allesch., (a), on conyza canadensis (l.) cronquist, gpu, dune, 30 aug. 2012, leg. & det. mrm. chaetosphaeria myriocarpa (fr.) c. booth, (t), on old stromata of hypoxylon fuscum (on corylus avellana log), gpu, deciduous thicket, 27 aug. 2013, leg. & det. js. cladosporium herbarum (pers.) link s. l., (a), on quercus robur l., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. md & es; on salix sp., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. md, det. md & es; on silene vulgaris (moench) garcke, opu, sośnia, mixed forest edge, 28 aug. 2013, leg. md, det. md & es. cladosporium sp., (a), on acer pseudoplatanus l., opu, góra skobla, deciduous thicket, 27 aug. 2013, leg. es, det. md & es; on corylus avellana l., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. md & es; on plantago lanceolata l., biały grąd, deciduous thicket, 27 aug. 2013, leg. md, det. md & es. coleosporium tussilaginis (pers.) lév. s. l., (iii, iii), on campanula trachelium l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; gpu, mixed forest, path, 29 aug. 2012, leg. & det. mrm; on melampyrum polonicum (p.b.) soó, gpu, deciduous forest edge, 30 aug. 2012, leg. & det. mrm; on melampyrum nemorosum l., opu, sośnia, scots pine forest, roadside, 28 aug. 2013, leg. & det. mrm. #colletotrichum circinans (berk.) voglino, (a), on paris quadrifolia l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es. colletotrichum dematium (pers.) grove, (a), on convallaria majalis l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es. colletotrichum sp., (a), on acer negundo l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es; on paris quadrifolia l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es. colletotrichum veratrina (ellis & everh.) treigiene, (a), on geranium palustre l., kpu, t-c, 28 aug. 2012, leg. mrm, det. mwk. coniothyrium olivaceum bonord. [= microsphaeropsis olivacea (bonord.) höhn.], (a), on cerasus avium (l.) moench, opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es, rev. mrm; on corylus avellana l., gpu, mixed forest, 29 aug. 2012, leg. & det. mrm; on dianthus carthusianorum l., gpu, dune, 27 aug. 2013, leg. mrm, det. mwk; on peucedanum cervaria (l.) lapeyr., gpu, deciduous thicket, small glade, 27 aug. 2013, leg. & det. mrm. 7 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park daldinia concentrica (bolton) ces. & de not. s. l., (t), on alnus glutinosa (l.) gaertn. log, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. js. daldinia decipiens wollw. & m. stadler, (t), on alnus glutinosa (l.) gaertn. log, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. tomasz leski, det. js. diatrype bullata (hoffm.) fr., (t), on corylus avellana l. branch, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. js; on salix sp. branch, opu, sośnia, ce-a, 28 aug. 2013, leg. & det. js. diatrype stigma (hoffm.) fr., (t), on alnus glutinosa (l.) gaertn. branch, opu, sośnia, ce-a, 28 aug. 2013, leg. & det. js; on corylus avellana l. branch, gpu, t-c, 27 aug. 2013, leg. & det. js. diatrypella favacea (fr.) ces. & de not., (t), on betula pendula roth branch, gpu, vu-b, 27 aug. 2013, leg. & det. js; same locality and date, leg. & det. małgorzata stasińska; opu, sośnia, deciduous thicket, 28 aug. 2013, leg. & det. js; on corylus avellana l. fallen branch, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. js. diatrypella quercina (pers.) cooke, (t), on quercus robur l. branch, gpu, t-c, 27 aug. 2013, leg. & det. js. diplocarpon earlianum (ellis & everh.) f.a. wolf [= marssonina fragariae (lib.) kleb.], (a), on comarum palustre l., gpu, willow thicket, 29 aug. 2012, leg. mrm, det. mwk. discosia artocreas (tode) fr., (a), on lysimachia thyrsiflora l., gpu, willow thicket, 29 aug. 2012, leg. & det. mrm; on oenothera biennis l. s. l., gpu, deciduous thicket, small glade, 27 aug. 2013; on peucedanum cervaria (l.) lapeyr., gpu, deciduous thicket, small glade, 27 aug. 2013, leg. & det. mrm; on populus tremula l., gpu, mixed forest, 29 aug. 2012, leg. & det. mrm; on quercus robur l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. & det. es. discosia fitzpatrickii vanev, (a), on quercus robur l., gpu, vu-p, 29 aug. 2012, leg. & det. mrm. discosia kabati vanev, (a), on sorbus aucuparia l. emend. hedl., gpu, dune edge, poor grassland, 30 aug. 2012, leg. & det. mrm. #discosia minuta ces., (a), on vaccinium myrtillus l., gpu, vu-p, 29 aug. 2012, leg. mrm, det. mrm & mwk; on vaccinium uliginosum l., gpu, vu-p, 29 aug. 2012, leg. mrm, det. mwk; on vaccinium vitis-idaea l., gpu, vu-p, 29 aug. 2012, leg. & det. mrm. discosia potentillae tehon, (a), on filipendula ulmaria (l.) maxim., gpu, willow thicket near catwalk, 29 aug. 2012, leg. & det. mrm. discosia pulmonariae vanev, (a), on pulmonaria officinalis l., gpu, deciduous forest, 29 aug. 2012, leg. & det. mrm. discosia quercicola de not., (a), on corylus avellana l., gpu, mixed forest, 29 aug. 2012, leg. & det. mrm. discosia sp., (a), on epilobium adnatum griseb., opu, góra skobla, deciduous thicket, 26 aug. 2013, leg. md, det. md & es. 8 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park #discostroma callistemonis (h.j. swart) sivan. [= seimatosporium kriegerianum (bres.) morgan-jones & b. sutton], (a), on chamaenerion angustifolium (l.) scop., wpu, carska droga near grobla honczarowska, roadside, 26 aug. 2013, leg. & det. mrm; on epilobium adnatum griseb., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es. #discula campestris (pass.) arx, (a), on acer platanoides l., opu, sośnia, village, 29 aug. 2013, leg. mw, det. mrm. epicoccum nigrum link, (a), on acer pseudoplatanus l., opu, góra skobla, deciduous forest, 26 aug. 2013, leg. es, det. md & es; on humulus lupulus l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; on salix sp., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. md, det. md & es. erysiphe adunca (wallr.) fr., (t), on salix aurita l., gpu, willow thicket, 29 aug. 2012, leg. & det. mrm. erysiphe alphitoides (griffon & maubl.) u. braun & s. takam., (a,t), on quercus robur l., gpu, vu-p, 29 aug. 2012, leg. & det. mrm; opu, biały grąd, deciduous forest, 27 aug. 2013, leg. es, det. md & es; góra skobla, mixed forest, 27 aug. 2013, leg. es, det. md & es; on quercus rubra l., (a), opu, carska droga, catwalk, 26 aug. 2013, leg. mrm & natalia michalska, det. mrm. erysiphe aquilegiae var. ranunculi (grev.) r.y. zheng & g.q. chen, (a), on ranunculus repens l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm; on ranunculus sp., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; on thalictrum aquilegiifolium l., opu, biały grąd, edge of causeway, 27 aug. 2013, leg. md, det. md & es. erysiphe berberidis dc., (a), on berberis vulgaris l., opu, góra skobla, deciduous forest edge, 27 aug. 2013, leg. md, det. md & es. erysiphe cynoglossi (wallr.) u. braun, (a), on pulmonaria officinalis l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. erysiphe divaricata (wallr.) schltdl., (t), on frangula alnus mill., gpu, deciduous thicket, 27 aug. 2013, leg. & det. mrm; opu, biały grąd, thicket close to mineral island (“grądzik”), 27 aug. 2013, leg. mw, det. mrm. erysiphe euonymi dc., (t), on euonymus europaea l., opu, sośnia, edge of ash-alder forest (fraxino-alnetum), close to dune, 28 aug. 2013, leg. & det. mrm; opu, sośnia, ce-a, close to dune, 29 aug. 2013, leg. & det. mrm. erysiphe friesii (lév.) u. braun & s. takam., (t), on rhamnus cathartica l., opu, sośnia, mixed forest near dune, 28 aug. 2013, leg. mw, det. mrm. erysiphe heraclei dc., (a), on heracleum sphondylium l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; same locality and date, (t), leg. & det. mrm; on peucedanum cervaria (l.) lapeyr., (t), gpu, deciduous thicket, small glade, 28 aug. 2013, leg. & det. mrm. erysiphe howeana u. braun, (a), on oenothera biennis l. s. l., gpu, dune, 27 aug. 2013, leg. & det. mrm. erysiphe ornata var. europaea (u. braun) u. braun & s. takam., (t), on betula pendula roth, gpu, vu-p, 29 aug. 2012, leg. & det. mrm; on betula pubescens ehrh., gpu, vu-p, 29 aug. 2012, leg. & det. mrm; opu, sośnia, scots pine forest, 28 aug. 2013, leg. & det. mrm; same locality and date, leg. es, det. md & es. 9 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park erysiphe polygoni dc., (a), on rumex acetosella l., gpu, dune, 27 aug. 2013, leg. & det. mrm. erysiphe sp., (a), on syringa vulgaris l., opu, sośnia, village, near house, 28 aug. 2013, leg. & det. mrm. erysiphe tortilis (wallr.) link, (t), on cornus sp., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. md & es. erysiphe trifolii grev., (a), on melilotus officinalis (l.) pall., bpu, grobla honczarowska causeway, 26 aug. 2013, leg. md, det. md & es. erysiphe urticae (wallr.) s. blumer on urtica dioica l., opu, sośnia, mixed forest edge, 28 aug. 2013, leg. md, det. md & es. erysiphe vanbruntiana (w.r. gerard) u. braun & s. takam., (a), on sambucus racemosa l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es. erysiphe viburni duby [= microsphaera sparsa howe], (t), on viburnum opulus l., opu, sośnia, deciduous forest near dune, 28 aug. 2013, leg. & det. mrm. eutypella cerviculata (fr.) sacc., (t), on alnus glutinosa (l.) gaertn. branch, opu, sośnia, ce-a, 28 aug. 2013, leg. & det. js. #exobasidium pachysporum nannf., (t), on vaccinium uliginosum l., bbpn buffer zone, 2 km sw osowiec-twierdza village, transitional bog, vu-b, 28 aug. 2013, leg. małgorzata stasińska, det. mrm. exobasidium vaccinii (fuckel) woronin, (t), on vaccinium vitis-idaea l., opu, dziegciorka, close to road 668 (from osowiec to sośnia), mixed forest, 28 aug. 2013, leg. mw, det. mrm. #fusicladium betulae aderh., (a), on betula pendula roth, gpu, catwalk, 27 aug. 2013, leg. & det. mrm. gloeosporium sp., (a), on acer negundo l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es; on quercus robur l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es. gnomoniopsis comari (p. karst.) sogonov [= zythia fragariae laib.], (a), on potentilla argentea l. (det. dr. jeremi kołodziejek), kpu, t-c, glade, 28 aug. 2012, leg. & det. mrm. golovinomyces sordidus (l. junell) v.p. heluta, (t), on plantago major l., opu, góra skobla, deciduous forest edge, 27 aug. 2013, leg. md, det. md & es. gymnosporangium cornutum arthur ex f. kern, (0, i), on sorbus aucuparia l. emend. hedl., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es; sośnia, dune, 28 aug. 2013, leg. natalia michalska, det. mrm; same locality and date, leg. md, det. md & es. gymnosporangium sabinae (dicks.) g. winter, (0, i), on pyrus communis l., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. md & es. helminthosphaeria clavariarum (desm.) fuckel, (t), on clavulina sp., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. błażej gierczyk. 10 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park hendersonia viciae-fabae savelli, (a), on vicia sp., gpu, edge of dune, 27 aug. 2013, leg. & det. mrm. hyaloperonospora niessliana (berl.) constant. [= peronospora niessliana berl.], (a), on alliaria petiolata (m. bieb.) cavara & grande, kpu, t-c, 28 aug. 2012, leg. mrm, det. mwk; opu, góra skobla, deciduous forest edge, 27 aug. 2013, leg. es, det. md & es. #hypomyces rosellus (alb. & schwein.) tul. & c. tul., (t), on wood of deciduous tree, opu, sośnia, mixed forest, 28 aug. 2013, leg. marcin pietras, det. anna kujawa, błażej gierczyk & js. hypoxylon fragiforme (pers.) j. kickx, (t), on alnus glutinosa (l.) gaertn. branch, opu, sośnia, ce-a, 28 aug. 2013, leg. & det. js. hypoxylon fuscum (pers.) fr., (t), on branches of acer platanoides l. and tilia cordata mill., gpu, t-c, 27 aug. 2013, leg. & det. js; on alnus glutinosa (l.) gaertn. branch, gpu, ce-a, 27 aug. 2013,leg. & det. js. hypoxylon howeanum peck., (t), on corylus avellana l. fallen branch, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. js. #lasiosphaeris hirsuta (fr.) a.n. mill. & huhndorf, (t), on alnus glutinosa (l.) gaertn. rotten branch, opu, sośnia, ce-a, 28 aug. 2013, leg. & det. js. leptosphaeria sp., (t), on acer pseudoplatanus l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es. leptotrochila ranunculi (fr.) schüepp, (t), on ranunculus acris l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; same locality and date, leg. mrm & natalia michalska, det. mrm. #leveillula helichrysi v.p. heluta & simonyan, (t), on helichrysum arenarium (l.) moench, gpu, dune, 27 aug. 2013, leg. & det. mrm. notes. the species reported at the same locality in bbnp in 2013 as leveillula taurica (lév.) arnaud [1]. also reports of majewski from świnoujście, drezdenko, zgłowiączka near włocławek, listed by sałata [42] as l. taurica, most probably belong to the same species. lophodermium pinastri (schrad.) chevall., (t), on juniperus communis l., opu, sośnia, mixed forest, 28 aug. 2013, leg. es, det. md & es. melampsora epitea thüm., (ii), on salix aurita l., gpu, willow thicket, 29 aug. 2012, leg. & det. mrm. melampsora laricis-pentandrae kleb., (ii), on salix sp., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es. melampsora populnea (pers.) p. karst., (ii, iii), on populus nigra l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; same locality and date, leg. & det. mrm; on populus tremula l., gpu, vu-p, 29 aug. 2012, leg. & det. mrm; same host, deciduous thicket, 27 aug. 2013, leg. & det. mrm. melampsoridium betulinum (pers.) kleb., (ii), on betula pendula roth, gpu, vu-p, 29 aug. 2012, leg. & det. mrm. #monilinia padi (woronin) honey [= monilia linhartiana sacc.], (a), on cerasus avium (l.) moench, opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. 11 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park md & es, rev. mrm; on padus serotina (ehrh.) borkh., opu, sośnia, scots pine forest with padus serotina thicket, 28 aug. 2013, leg. & det. mrm. monochaetia monochaeta (desm.) allesch., (a), on quercus robur l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es. mycosphaerella millegrana (cooke) j. schröt. [= passalora microsora (sacc.) u. braun], (a), on tilia cordata mill., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. mrm. mycosphaerella osborniae d. hawksw. & sivanesan, (t), on artemisia vulgaris, kpu, t-c, road edge, 28 aug. 2012, leg. & det. mrm. naohidemyces vaccinii (jørst.) s. sato, katsuya & y. hirats. ex vanderweyen & fraiture [= pucciniastrum vaccinii jørst.], (ii), on vaccinium uliginosum l., bbpn buffer zone, 2 km sw osowiec-twierdza village, transitional bog, vu-b, 28 aug. 2013, leg. małgorzata stasińska, det. mrm. nectria cinnabarina (tode) fr., (t), on betula pendula roth branches, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. js. nemania aenea (nitschke) pouzar, (t), on rotten branch of salix fragilis l. submerged in stagnant water, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. js. nemania serpens (pers.) gray, (t), on carpinus betulus l. log, gpu, t-c, 27 aug. 2013, leg. & det. js. neoerysiphe galeopsidis (dc.) u. braun, (a), on galeobdolon luteum huds., gpu, t-c, 27 aug. 2013, leg. & det. mrm; on galeopsis tetrahit l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es. oidium sp., (a), on vicia sp., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es. ophiognomonia intermedia (rehm) sogonov [= discula betulina (westend.) arx], (a), on betula pendula roth, gpu, vu-p, 29 aug. 2012, leg. & det. mrm; catwalk, 27 aug. 2013, leg. & det. mrm; on betula pubescens ehrh., gpu, catwalk, 27 aug. 2013, leg. & det. mrm. passalora ariae (fuckel) u. braun & crous, (a), on sorbus aucuparia l. emend. hedl., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. passalora comari (peck) u. braun [= cercospora comari peck], (a), on comarum palustre l., gpu, willow thicket, 29 aug. 2012, leg. mrm, det. mwk. passalora ferruginea (fuckel) u. braun & crous, (a), on artemisia vulgaris l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm; kpu, t-c, edge of the road, 28 aug. 2012, leg. & det. mrm; opu, góra skobla, deciduous forest, 27 aug. 2013, leg. md, det. md & es. #passalora heterospora (höhn.) höhn., (a), on epilobium roseum schreb., wpu, carska droga near grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. #passalora minutissima (desm.) u. braun & crous, (a), on geranium palustre l., opu, góra skobla, deciduous forest edge, 26 aug. 2013, leg. es, det. md & es, rev. mrm. 12 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park passalora murina (ellis & kellerm.) u. braun & crous, (a), on viola sp., gpu, mixed forest, 29 aug. 2012, leg. mrm, det. mwk. periconia byssoides pers., (a), on lonicera xylosteum l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. md & es; opu, góra skobla, deciduous forest edge, 27 aug. md, det. md & es; on viburnum opulus l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es. periconia cookei mason & m.b. ellis, (a), on polygonatum odoratum (mill.) druce, gpu, t-c, 27 aug. 2013, leg. & det. mrm. #peronospora agrimoniae syd., (a), on agrimonia eupatoria l. s. l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. peronospora knautiae fuckel, (a), on knautia arvensis (l.) j.m. coult., opu, sośnia, deciduous forest edge, 28 aug. 2013, leg. & det. es & md. peronospora sordida berk., (a), on scrophularia nodosa l., gpu, deciduous thicket, 27 aug. 2013, leg. & det. mrm. phaeobotryosphaeria visci (kalchbr.) a.j.l. phillips & crous [= sphaeropsis visci (alb. & schwein.) sacc.], (a), on viscum album l., gpu, catwalk, 27 aug. 2013, leg. & det. mrm. phaeosphaeria inclusa shoemaker & c.e. babc., (t), on polygonatum odoratum (mill.) druce, gpu, t-c, 27 aug. 2013, leg. & det. mrm. phaeosphaeria sp., (t), on thelypteris palustris schott, bpu, długa luka overpass, sedge patch, 26 aug. 2013, leg. & det. mrm. phloeospora aceris (lib.) sacc., (a), on acer platanoides l., opu, sośnia, village, 29 aug. 2013, leg. mw, det. mrm; on acer pseudoplatanus l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es. phoma cf. macrostoma mont., (a), on fraxinus excelsior l., gpu, mixed forest, 29 aug. 2012, leg. & det. mrm. phoma glomerata (corda) wollenw. & hochapfel, (a), on erysiphe euonymi (on euonymus europaea), opu, sośnia, ce-a, close to dune, 29 aug. 2013, leg. mw, det. mrm; on erysiphe divaricata (on frangula alnus), gpu, deciduous thicket, 27 aug. 2013, leg. & det. mrm. phoma sect. heterospora boerema, gruyter & noordel., (a), on dianthus carthusianorum l., gpu, dune, 27 aug. 2013, leg. mrm, det. mwk. phomatospora dinemasporium j. webster [= dinemasporium strigosum (pers.) sacc.], (a), on corynephorus canescens (l.) p. beauv., gpu, dune, 20 aug. 2012, leg. mrm, det. mwk & mrm. phomopsis amygdaliana canonaco, (a), on padus serotina (ehrh.) borkh., opu, sośnia, scots pine forest with padus serotina thicket, 28 aug. 2013, leg. & det. mrm. phragmidium potentillae (pers.) p. karst., (ii, iii), on potentilla argentea l., gpu, dune, 27 aug. 2013, leg. mrm, det. mwk. phragmidium tuberculatum jul. müll., (ii, iii), on rosa canina l., opu, sośnia, mixed forest edge, 28 aug., leg. es, det. md & es. 13 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park phyllactinia guttata (wallr.) lév. s. l., (a,t), on betula pendula roth, gpu, catwalk, 27 aug. 2013, leg. & det. mrm; on corylus avellana l., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. md & es. #phyllosticta pyrolae ell. & everh., (a), on orthilia secunda (l.) house, opu, sośnia, scots pine forest, near dune, 28 aug. 2013, leg. & det. mrm. phyllosticta tambowiensis bubák & serebrian., (a), on acer negundo l., opu, sośnia, mixed forest, 28 aug. 2013, leg. es, det. md & es. #pilidium concavum (desm.) höhn., (a), on epilobium adnatum griseb., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es. #plenodomus visci (sacc.) gruyter, aveskamp & verkley [= plectophomella visci (sacc.) moesz], (a), on viscum album l., gpu, catwalk, 27 aug. 2013, leg. & det. mrm. podosphaera aphanis (wallr.) u. braun & s. takam. [= sphaerotheca alchemillae (j. steiner) erikss.], (a,t), on alchemilla sp., opu, biały grąd, edge of causeway, 27 aug. 2013, leg. & det. es; on filipendula ulmaria (l.) maxim., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es. podosphaera balsaminae (wallr.) u. braun & s. takam., (a), on impatiens noli-tangere l., bpu, grobla honczarowska causeway, 26 aug. 2013, leg. es, det. md & es; same locality and date, leg. & det. mrm. podosphaera fusca (fr.) u. braun & shishkoff, (a,t), on conyza canadensis (l.) cronquist, gpu, dune, 27 aug. 2013, leg. & det. mrm; on erigeron annuus (l.) pers., opu, sośnia, mixed forest edge, 28 aug. 2013, leg. & det. es; on melampyrum nemorosum l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm; on melampyrum sp., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; on taraxacum officinale f.h. wigg., opu, biały grąd, edge of causeway, 27 aug. 2013, leg. es, det. md & es; sośnia, mixed forest edge, 28 aug. 2013, leg. & det. es. podosphaera major (juel) s. blumer [= p. myrtillina kunze var. major juel], (t), on vaccinium uliginosum l., gpu, vu-p, 29 aug. 2012, leg. mrm, det. mwk; bbpn buffer zone, 2 km sw osowiec-twierdza village, transitional bog, vu-b, 28 aug. 2013, leg. małgorzata stasińska, det. mrm. podosphaera myrtillina kunze, (t), on vaccinium myrtillus l., gpu, vu-p, 29 aug. 2012, leg. mrm, det. mwk. puccinia arenariae (schumach.) j. schröt., (ii, iii), on melandrium album (mill.) garcke, opu, góra skobla, deciduous forest edge, 27 aug. 2013, leg. md, det. md & es; bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; on moehringia trinervia (l.) clairv., gpu, mixed forest, 29 aug. 2012, leg. & det. mrm; kpu, t-c, 28 aug. 2012, leg. natalia michalska, det. mrm; on stellaria holostea l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es. puccinia argentata (schultz) g. winter [= p. impatientis ficinus & c. schubad], (ii, iii), on impatiens noli-tangere l., bpu, grobla honczarowska causeway, 26 aug. 2013, leg. es, det. md & es; same locality and date, leg. & det. mrm. puccinia chaerophylli purton, (ii, iii), on chaerophyllum temulum l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. 14 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park puccinia hieracii (röhl.) h. mart., (ii), on taraxacum officinale f.h. wigg., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. md & es. puccinia menthae pers., (i, ii), on mentha arvensis l., opu, biały grąd, edge of causeway, 27 aug. 2013, leg. es, det. md & es. puccinia oreoselini (f. strauss) körn., (iii), on peucedanum cervaria (l.) lapeyr., gpu, deciduous forest edge, 27 aug. 2013, leg. & det. mrm. puccinia phragmitis tul. [= p. phragmitis (schumach.) körn]., (ii, iii), on phragmites australis (cav.) trin. ex steud., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es. puccinia polygoni-amphibii pers. [= p. polygoni alb. & schwein.], (ii, iii), on fallopia convolvulus (l.) á. löve, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. md & es; gpu, mixed forest, 29 aug. 2012, leg. mrm, det. mrm & mwk; dune, 27 aug. 2013, leg. & det. mrm; opu, biały grąd, edge of causeway, 27 aug. 2013, leg. md, det. md & es. puccinia punctata link, (ii, iii), on galium schultesii vest, gpu, deciduous thicket, 27 aug. 2013, leg. & det. mrm; on galium sp., opu, sośnia, mixed forest edge, 28 aug. 2013, leg. md, det. md & es. puccinia punctiformis (f. strauss) röhl., (ii, iii), on cirsium oleraceum (l.) scop., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es. puccinia tanaceti dc. s. l., (ii,iii), on artemisia vulgaris l., kpu, t-c, road edge, 28 aug. 2012, leg. & det. mrm; opu, góra skobla, deciduous forest, 27 aug. 2013, leg. md, det. md & es; sośnia, mixed forest edge, 28 aug. 2013, leg. md, det. md & es. pyrenochaeta sp., (a), on vaccinium vitis-idaea l., gpu, vu-p, 29 aug. 2012, leg. mrm, det. mwk. ramularia agrestis sacc., (a), on viola arvensis murray, gpu, dune, 27 aug. 2013, leg. & det. mrm. ramularia aromatica (sacc.) höhn., (a), on acorus calamus l., opu, biały grąd, edge of causeway, ditch, 27 aug. 2013, leg. es, det. md & es. ramularia carneola (sacc.) nannf. [= r. scrophulariae fautrey & roum.], (a), on scrophularia nodosa l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; same locality and date, leg. & det. mrm; gpu, deciduous thicket, 27 aug. 2013, leg. & det. mrm. ramularia chaerophylli ferraris, (a), on chaerophyllum temulum l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. ramularia chamaedryos (lindr.) gunnerb., (a), on veronica chamaedrys l., bpu, grobla honczarowska causeway,deciduous thicket, 26 aug. 2013, leg. & det. mrm. ramularia geranii (westend.) fuckel var. geranii, (a), on geranium palustre l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. ramularia grevilleana (tul.) jørst. var. grevilleana, (a), on potentilla argentea l. (det. dr jeremi kołodziejek), gpu, edge of dune, 27 aug. 2013, leg. & det. mrm. 15 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park ramularia inaequalis (preuss) u. braun, (a), on cichorium intybus l., opu, sośnia, mixed forest edge, 28 aug. 2013, leg. es, det. md & es. ramularia lysimachiae thüm., (a), on lysimachia thyrsiflora l., gpu, vu-p, 29 aug. 2012, leg. & det. mrm. ramularia moehringiae lindr., (a), on moehringia trinervia (l.) clairv., gpu, mixed forest, 29 aug. 2012, leg. & det. mrm. ramularia rhabdospora (berk. & broome) nannf., (a), on plantago lanceolata l., opu, biały grąd, edge of causeway, 27 aug. 2013, leg. es, det. md & es. #ramularia rhaetica (sacc. & g. winter) jaap, (a), on peucedanum cervaria (l.) lapeyr., gpu, deciduous thicket, small glade, 27 aug. 2013, leg. & det. mrm. ramularia simplex pass., (a), on ranunculus repens l., bpu, grobla honczarowska, deciduous thicket, 26 aug. 2013, leg. & det. mrm. ramularia urticae ces., (a), on urtica dioica l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; opu, biały grąd, edge of causeway, 27 aug. 2013, leg. md, det. md & es. ramularia variabilis fuckel, (a), on verbascum nigrum l., gpu, dune, 27 aug. 2013, leg. & det. mrm. rhytisma acerinum (pers.) fr. [= melasmia acerina lév.], (a), on acer platanoides l., opu, osowiec-twierdza village, near the headquarters of bbnp, 28 aug. 2013, leg. & det. jc; sośnia, village, 29 aug. 2013, leg. mw, det. mrm; on acer pseudoplatanus l., opu, góra skobla, 27 aug. 2013, leg. es, det. md & es. #sarcopodium circinatum ehrenb., (a), on geranium robertianum l., gpu, mixed forest, 29 aug. 2012, leg. mrm, det. mwk; on mercurialis perennis l., gpu, mixed forest, 29 aug. 2012, leg. mrm, det. mwk. sawadaea bicornis (wallr.) homma, (t), on acer pseudoplatanus l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. es, det. md & es. sawadaea tulasnei (fuckel) homma, (t), on acer pseudoplatanus l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. & det. md & es. seimatosporium foliicola (berk.) shoemaker, (a), on crataegus monogyna jacq., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. md, det. md & es. septoria aegopodii desm., (a), on aegopodium podagraria l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm; gpu, catwalk, 27 aug. 2013, leg. natalia michalska, det. mrm; kpu, t-c, 28 aug. 2012, leg. & det. mrm; on peucedanum oreoselinum (l.) moench, gpu, edge of dune, 30 aug. 2012, leg. mrm, det. mwk. #septoria alni sacc., (a), on alnus glutinosa (l.) gaertn., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. septoria brissaceana sacc. & letell., (a), on lythrum salicaria l., opu, biały grąd, edge of causeway, 27 aug. 2013, leg. md, det. md & es. septoria capraeae westend., (a), on salix sp., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. md & es. 16 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park septoria chelidonii desm., (a), on chelidonium majus l., opu, biały grąd, edge of causeway, 27 aug. 2013, leg. md, det. md & es; góra skobla, deciduous thicket, 27 aug. 2013, leg. md, det. md & es. septoria convolvuli desm., (a), on convolvulus arvensis l., gpu, dune slope, 30 aug. 2012, leg. & det. mrm. septoria epilobii westend., (a), on epilobium roseum schreb., wpu, carska droga near grobla honczarowska causeway, deciduous thickets, 26 aug. 2013, leg. & det. mrm; associated with immature mycosphaerella sp. ascomata. septoria galeopsidis westend., (a), on galeopsis tetrahit l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; opu, góra skobla, deciduous forest, 27 aug. 2013, leg. md, det. md & es. #septoria jasiones (bres.) grove, (a), on jasione montana l., gpu, dune, 27 aug. 2013, leg. & det. mrm. septoria lychnidis desm. [= caryophylloseptoria lychnidis (desm.) verkley, quaedvlieg & crous], (a), on melandrium album (mill.) garcke, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; opu, sośnia, mixed forest edge, 28 aug. 2013, leg. md, det. md & es. septoria lysimachiae westend., (a), on lysimachia thyrsiflora l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; same locality and date, leg. & det. mrm; gpu, willow thicket, 29 aug. 2012, leg. & det. mrm; opu, sośnia, mixed forest near dune, 28 aug. 2013, leg. & det. mrm. septoria oenotherae westend., (a), on oenothera biennis l. s. l., gpu, deciduous thicket, small glade, 27 aug. 2013, leg. & det. mrm; same locality, dune, 27 aug. 2013, leg. natalia michalska, det. mrm; sośnia, roadside, 28 aug. 2013, leg. md, det. md & es. septoria oreoselini (lasch) sacc., (a), on peucedanum cervaria (l.) lapeyr., gpu, deciduous thicket, small glade, 27 aug. 2013, leg. & det. mrm. septoria polygonorum desm., (a), on polygonum persicaria l., opu, biały grąd, edge of causeway, 27 aug. 2013, leg. es, det. md & es. septoria pyricola desm., (a), on pyrus communis l., opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. es, det. md & es. septoria scabiosicola desm., (a), on knautia arvensis (l.) j.m. coult., opu, sośnia, mixed forest edge, 28 aug. 2013, leg. es, det. md & es; gpu, egde of dune, 29 aug. 2013, leg. & det. mrm. #septoria tabacina died., (a), on artemisia vulgaris l., kpu, t-c, edge of forest road, 28 aug. 2012, leg. & det. mrm. septoria virgaureae (lib.) desm., (a), on solidago virgaurea l., gpu, edge of dune, 30 aug. 2012, leg. & det. mrm. sphaceloma viburni jenkins & bitanc., (a), on viburnum opulus l., opu, sośnia, deciduous forest near dune, 28 aug. 2013, leg. & det. mrm. sphaerellopsis filum (biv.) b. sutton, (a), on melampsora epitea (on salix autira), gpu, willow thicket, 29 aug. 2012, leg. & det. mrm; on puccinia arenariae (on melandrium album), bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es, rev. mrm; on puccinia argentata (on impatiens noli-tangere), bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, 17 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park det. md & es; on puccinia menthae (on mentha arvensis), opu, biały grąd, edge of causeway, 27 aug. 2013, leg. es, det. md & es; on puccinia phragmitis (on phragmites australis), bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; on puccinia punctiformis (on cirsium oleraceum), bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. es, det. md & es; on uromyces geranii (on geranium palustre), bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. sphaerulina sp., (t), on euonymus europaea l., opu, sośnia, ce-a, close to edge of dune, 29 aug. 2013, leg. & det. mrm; on scrophularia nodosa l., gpu, deciduous thicket, 27 aug. 2013, leg. & det. mrm, associated with ramularia carneola. #stephanoma strigosum wallr., (a), on humaria hemisphaerica (f.h. wigg.) fuckel, opu, sośnia, mixed forest (pinus sylvestris, juniperus communis, populus tremula), 28 aug. 2013, leg. grażyna domian, det. mrm. notes. this species is known in poland only from four records in the early twentyfirst-century papers listed by mułenko et al. [32] as stephanoma strigosum and hypomyces strigosus (wallr.) schroeter. however, the species seems to be very common on the host throughout the country (m. ruszkiewicz-michalska, unpblished). #thedgonia bellocensis (c. massal. & sacc.) u. braun, (a), on verbascum nigrum l., gpu, vu-b, close to edge of dune, 27 aug. 2013, leg. & det. mrm. #thedgonia ligustrina (boerema) b. sutton, (a), bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. mrm. trichothecium roseum (pers.) link, (a), on acer negundo l., opu, góra skobla, deciduous forest, 27 aug. 2013, leg. & det. es. trichothecium sp., (a), on alliaria petiolata (m. bieb.) cavara & grande, kpu, t-c, 28 aug. 2012, leg. mrm, det. mwk; on vaccinium myrtillus l., gpu, vu-p, 29 aug. 2012, leg. mrm, det. mwk. triphragmium ulmariae (dc.) link (ii, iii) on filipendula almaria (l.) maxim., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; same locality and date, leg. & det. mrm. uromyces geranii (dc.) g.h. otth & wartm. (iii) on geranium palustre l., bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. md, det. md & es; same date, leg. & det. mrm. uromyces rumicis (schumach.) g. winter (iii) on rumex hydrolapathum huds., opu, biały grąd,edge of causeway, 27 aug. 2013, leg. & det. es. uromyces viciae-fabae (pers.) j. schröt. (ii, iii) on vicia cracca l., gpu, dune, 30 aug. 2012, leg. & det. mrm. valsa pini (alb. & schwein.) fr., (t), on pinus sylvestris l. branch, gpu, dune, 27 aug. 2013, leg. & det. js. venturia potentillae (wallr.) cooke [= coleroa potentillae (wallr.) g. winter], (a), on comarum palustre l., bpu, długa luka overpass, sedge patch, 26 aug. 2013, leg. mrm, det. mwk; gpu, willow thicket, 29 aug. 2012, leg. mrm, det. mwk. fungi associated with arthropods in total 16 species of entomopathogenic fungi were found as pathogens of different arthropods and 6 species were isolated from forest or meadow soil by means of the 18 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park galleria bait method or warcup dilution. nine entomophthoralean fungal species representing phylum entomophthoromycota were isolated in total, among them seven were identified as pathogens of insects, one (neozygites floridana) was isolated from spider mite and conidiobolus coronatus was detected from meadow soil by means of the bait method and from plant debris by dilution method. from among seven species of anamorphic hypocreales (ascomycota) affecting arthropods in investigated parts of the bbnp in 2013, two have been recognized as pathogens of spiders (gibellula leiopus and g. pulchra), two (beauveria bassiana and isaria farinosa) were isolated from different unidentified insects in forest litter and three were pathogenic to mites (hirsutella danubiensis, h. kirchneri, h. thompsonii). in addition, from forest or meadow soil the entomopathogenic fungi isaria farinosa, i. fumosorosea, lecanicillium lecanii, lecanicillium sp. and metarhizium anisopliae were isolated by means of the galleria bait or dilution method. thirteen fungal species associated with arthropods recorded for the first time in bbnp are marked with asterisk. especially interesting is a record of mycoses in hope aphid (phorodon humuli) population, caused simultaneously by three entomophthoralean fungal species: entomophthora planchoniana, pandora neoaphidis and zoophthora aphidis. there were very limited possibilities to enter deeper into wet meadows because of both-side digs filled with water along the paths and roads. some accessible insects on dig-border plants appeared healthy, including even big aphid colonies on salix cinerea leaves, entirely free of fungal pathogens. beauveria bassiana (bals.-criv.) vuill. on unidentified bug (heteroptera), opu, sośnia, in the litter of deciduous forest, 28 aug. 2013, leg. & det. ct; bpu, grobla honczarowska; opu, biały grąd, and in some surrounding forests, common in the forest litter and in sub-cortical insects feeding sites on adult bark beetles and accompanying them small predacious larvae and adults, as well as in the coniferous and mixed forest litter mostly on chrysomelid, curculionid and staphylinid beetles, leg. & det. sb. cladosporium sp. (thick-walled, smooth spores) on exuvia of aphids feeding on alnus glutinosa (l.) gaertn. and on verbascum nigrum l., opu, sośnia, village path, 28 aug. 2013, leg. natalia michalska, det. mrm. conidiobolus coronatus (costantin) a. batko, plant debris, dilution method, kopu, biele suchowolskie fen, sp-c, 15 oct. 2009, leg. ab, det. mw; meadow soil, galleria bait method, opu, biały grąd, pasture, 27 aug. 2013, leg. & det. ct. entomophthora muscae (cohn) fresen. on unidentified fly (diptera) attached to the mentha sp. leaves, opu, biały grąd, pasture, 27 aug. 2013, leg. & det. ct. entomophthora planchoniana cornu on hope aphids (phorodon humuli schrank) feeding on humulus lupulus l. leaves, opu, sośnia, mixed forest edge, 28 aug., leg. & det. ct & sb. gibellula leiopus (vuill. ex maubl.) mains on a spider on underside of corylus avellana l. leaf, opu, biały grąd, deciduous thicket, 27 aug. 2013, leg. & det. sb. gibellula pulchra cavara on spider from the family thomisidae, bpu, grobla honczarowska causeway, deciduous thicket, 26 aug. 2013, leg. & det. sb & ct. hirsutella danubiensis tkaczuk, bałazy & wegenst. on spider mite neotetranychus rubi (träg) feeding on rubus sp., opu, biały grąd, deciduous forest, 27 aug. 2013, leg. & det. ct. hirsutella kirchneri (rostrup) minter, brady & hall on eriophyid mites (abacarus sp.) feeding on grass leaves, opu, góra skobla, mid-forest clearing, 28 aug. 2013, leg. & det. ct. 19 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park hirsutella thompsonii f.e. fisher on eriophyid mites (abacarus sp.) feeding on grass leaves, opu, góra skobla, mid-forest clearing, 28 aug. 2013, leg. & det. ct. isaria farinosa (holm) brown & smith, soil, warcup method, kopu, biele suchowolskie fen, fdc, s-cn, sp-c, 20 jun. 2008, leg. ab, det. mw; fdc, 15 oct. 2009, leg. ab, det. mw; forest soil, galleria bait method, opu, biały grąd, mixed forest edge, 27 aug. 2013, leg. & det. ct; in forest litter on caterpillars and some small beetles, single specimens in all investigated localities, leg. & det. sb. isaria fumosorosea wize, forest soil, galleria bait method, opu, biały grąd, mixed forest edge, 27 aug. 2013, leg. & det. ct. lecanicillium lecani (zimm.) zare & w. gams, soil, dilution method, kopu, biele suchowolskie fen, fdc, sp-c, 20 jun. 2008, leg. ab, det. mw; m-a, 15 oct. 2009, leg. ab, det. mw. lecanicillium sp., forest soil, galleria bait method, opu, biały grąd, pasture, 27 aug. 2013, leg. & det. ct. metarhizium anisopliae (metschn.) sorokin s. l., soil, dilution method, kopu, biele suchowolskie fen, s-cn, 20 jun. 2008, leg. ab, det. mw; forest soil, galleria bait method, opu, biały grąd, mixed forest edge, 27 aug. 2013, leg. & det. ct. neozygites floridana (j. weiser & muma) remaud. & keller on spider mites neotetranychus rubi feeding on rubus sp., opu, biały grąd, deciduous forest, 27 aug. 2013, leg. & det. ct. neozygites parvispora (d.m. macleod & k.p. carl) remaud. & s. keller on unidentified thrips feeding on solanum sp. leaves, opu, biały grąd, pastures, 27 aug. 2013, leg. & det. ct. pandora neoaphidis (remaud. & hennebert) humber on hope aphids phorodon humuli schrank feeding on humulus lupulus leaves, opu, sośnia, deciduous forest, 28 aug. 2013, leg. & det. ct & sb. verticillium sp. on a relatively big plant-hopper (auchenorrhyncha), partly destroyed, opu, sośnia, mid-forest marsh, on aquatic vegetation, 28 aug. 2013, leg. & det. sb; contamined secondarily by cladosporium herbarum (pers.) link s. l. zoophthora aphidis (hoffm. ex fresen.) a. batko on hope aphids phorodon humuli feeding on humulus lupulus leaves, opu, sośnia, deciduous forest, 28 aug. 2013, leg. & det. ct & sb. zoophthora ichneumonis bałazy on unidentified adults of hymenoptera from the family ichneumonidae, attached to the leaves of different deciduous trees, midmeadow loose afforestation, biały grąd, 27 aug. 2013, leg. & det. ct & sb. zoophthora sp. on unidentified beetle (coleoptera) feeding on corylus avellana leaves, bpu, grobla honczarowska, wooded dike, 26 aug.2013, leg. & det. ct. fungi and dictyostelia isolated from soil and associated with plant debris and excrements in total 61 species belonging to kickxellomycotina, mortierellomycotina, mucoromycotina, zoopagomycotina and ascomycetous anamorphs (41 taxa) were isolated. additionally two cellular slime moulds (amebozoa, dictyostelia) were found. 48 taxa reported here were not recorded during the first inventory in bbnp. among isolated fungi five taxa were found for the first time in poland. candelabrum spinulosum – very distinctive species growing among sphagnum was recorded 20 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park in the same year in peat bog “torfy” near warsaw (unpublished data). one of the most interesting among recorded species was piptocephalis fimbriata that has not been recorded from poland to date [32]. it is known from japan, taiwan, uk and usa [43]. the genus helicocephalum comprises parasites of nematodes eggs. until now in poland helicocephalum species was recorded only once, on wet, decayed wood [32]. however, the studies dealing with helicocephalidaceae were not conducted in poland and there is no precise information about rarity of the genus. our specimen was nonculturable and its affinity to the species remained unresolved. also one spororomiaceae strain remained unidentified. several taxa from this family are rare in poland and the number of known records is limited [32,38,44]. trichoderma aggressivum, known as fungal pathogen [45], was previously recorded from poland from mushroom compost [46]. two other taxa: talaromyces funiculosus and t. rugulosus were not listed in the polish checklist of micromycetes [32] but they are certainly quite common in our country [47,48]. piptocephalis lepidula and syncephalis species with single known records in poland [32,49], seem to be relatively common on dung of mice, rats and boars (m. wrzosek, unpublished). absidia glauca hagem, soil, damp chamber, opu, biały grąd, meadow, 27 aug. 2013, leg. & det. mw. acremoniella atra (corda) sacc., (a), soil, warcup method, kopu, biele suchowolskie fen, fdc, sp-c, 15 oct. 2009, leg. & det. mw. aspergillus flavus link, (a), soil, dilution method, kopu, biele suchowolskie fen, m-a, 20 jun. 2008, leg. & det. mw. aspergillus niger tiegh., (a), soil, dilution method, kopu, biele suchowolskie fen, fdc, m-a, s-cn, sp-c, 20 jun. 2008, leg. & det. mw. bionectria solani (reinke & berthold) schroers [= gliocladium solani (harting) petch], (a), soil, dilution method, kopu, biele suchowolskie fen, fdc, 20 jun. 2008, leg. ab, det. mw. candelabrum spinulosum beverw., plant debris from peatbog, opu, osowiec, birch forest, 27 aug. 2013, leg. & det. mw. cephalotrichum nanum (ehrenb.) s. hughes [= doratomyces nanus (ehrenb.) f.j. morton & g. sm.], (a), soil, warcup method, kopu, biele suchowolskie fen, m-a, 20 jun. 2008, leg. ab, det. mw. cephalotrichum stemonitis (pers.) nees [= doratomyces stemonitis (pers.) f.j. morton & g. sm.], (a), soil, warcup method, kopu, biele suchowolskie fen, sp-c, 15 oct. 2009, leg. ab, det. mw. cladosporium cladosporioides (fresen.) g.a. de vries, (a), soil, warcup and dilution methods, kopu, biele suchowolskie fen, m-a, 20 jun. 2008, leg. ab, det. mw; m-a, s-cn, sp-c, 15 oct. 2009, leg. & det. mw. cladosporium herbarum (pers.) link, (a), soil, warcup and dilution methods, kopu, biele suchowolskie fen, fdc, m-a, sp-c, 20 jun. 2008, leg. & det. mw; s-cn, 15 oct. 2009, leg. & det. mw. dictyostelium mucoroides bref., soil, dilution method, kopu, biele suchowolskie fen, m-a, 20 jun. 2008 and 15 oct. 2009, leg. ab, det. mw. fusarium arthrosporioides sherb., (a), soil and plant debris, warcup method, opu, biały grąd, meadow, 27 aug. 2013, leg. & det. mw. 21 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park fusarium chlamydosporum wollenw. & reinking [= fusarium fusarioides (gonz. frag. & cif.) c. booth], (a), soil, dilution and warcup methods, kopu, biele suchowolskie fen, m-a, 15 oct. 2009, leg. ab, det. mw. fusarium oxysporum schltdl., (a), soil, dilution and warcup methods, kopu, biele suchowolskie fen, m-a, 20 jun. 2008, leg. ab, det. mw; sp-c, 15 oct. 2009, leg. ab, det. mw. fusarium sporotrichioides sherb., (a), soil, dilution and warcup methods, kopu, biele suchowolskie fen, m-a, s-cn, 20 jun. 2008, leg. ab, det. mw; m-a, 15 oct. 2009, leg. ab, det. mw. gilmaniella sp., (a), soil, warcup method, kopu, biele suchowolskie fen, m-a, 15 oct. 2009, leg. ab, det. mw. helicocephalum sp., (a), on wet spruce wood, non-culturable, opu, carska droga, edge of road, 31 aug. 2013, leg. & det. mw. hirsutella rhossiliensis minter & b.l. brady, (a), soil, warcup method, kopu, biele suchowolskie fen, m-a, 15 oct. 2009, leg. ab, det. mw. humicola fuscoatra traaen, (a), soil, warcup method, kopu, biele suchowolskie fen, m-a, s-cn, 20 jun. 2008, leg. ab, det. mw; same plant communities, 15 oct. 2009, leg. ab, det. mw. microascus brevicaulis s.p. abbott [= scopulariopsis brevicaulis (sacc.) bainier], (a), soil, dilution method, kopu, biele suchowolskie fen, fdc, 15 oct. 2009, leg. & det. mw. mortierella bisporalis (thaxt.) björl., soil, damp chamber, opu, sośnia, ce-a, edge of dune, 29 aug. 2013, leg. & det. mw. mucor circinelloides tiegh., soil, dilution method, opu, sośnia, path in village, 28 aug. 2013, leg. & det. mw. mucor hiemalis wehmer, soil, warcup and dilution methods, opu, góra skobla, dune, 27 aug. 2013, leg. & det. mw; sośnia, ce-a, close to edge of dune, 29 aug. 2013, leg. & det. mw. mucor moelleri (vuill.) lendn. [= zygorhynchus moelleri vuill.], mineral soil, dilution method, opu, sośnia, dune, 29 aug. 2013, leg. & det. mw. mucor piriformis a. fisch., soil and plant debris, damp chamber method, opu, sośnia, path in village, 28 aug. 2013, leg. & det. mw. mucor racemosus fresen., soil, warcup and dilution methods, opu, góra skobla, dune, 27 aug. 2013, leg. & det. mw; sośnia, ce-a close to edge of dune, 29 aug. 2013, leg. & det. mw. mucor ramosissimus samouts., soil, dilution method, opu, sośnia, dune, 29 aug. 2013, leg. & det. mw. paecilomyces carneus (duché & r. heim) a.h.s. br. & g. sm., (a), soil, dilution method, kopu, biele suchowolskie fen, m-a, 15 oct. 2009, leg. ab, det. mw. paecilomyces sp., (a), soil, dilution and warcup methods, kopu, biele suchowolskie fen, fdc, sp-c, 20 jun. 2008, leg. ab, det. mw; fdc, m-a, sp-c, 15 oct. 2009, leg. ab, det. mw. 22 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park paecilomyces variotii bainier, (a), soil, warcup method, kopu, biele suchowolskie fen, m-a, 20 jun. 2008, leg. & det. mw. penicillium aurantiogriseum dierckx, (a), soil, dilution method, kopu, biele suchowolskie fen, s-cn, 20 jun. 2008, leg. & det. mw; sp-c, 15 oct. 2009, leg. & det. mw. penicillium brevicompactum dierckx, (a), soil, dilution method, kopu, biele suchowolskie fen, s-cn, sp-c, 15 ct. 2009, leg. & det. mw. penicillium chrysogenum var. chrysogenum thom, (a), soil, dilution and warcup methods, kopu, biele suchowolskie fen, fdc, m-a, s-cn, sp-c, 20 jun. 2008, leg. ab, det. mw. penicillium decumbens thom, (a), soil, dilution method, kopu, biele suchowolskie fen, sp-c, 20 jun. 2008, leg. ab, det. mw; m-a, sp-c, 15 oct. 2009, leg. ab, det. mw. penicillium miczynskii k.m. zaleski [= penicillium soppi k.m. zaleski], (a), soil, dilution method, kopu, biele suchowolskie fen, fdc, m-a, s-cn, sp-c, 20 jun. 2008, leg. ab, det. mw; m-a, sp-c, 15 oct. 2009, leg. ab, det. mw. penicillium oxalicum currie & thom, (a), soil, dilution method, kopu, biele suchowolskie fen, s-cn, 20 jun. 2008, leg. ab, det. mw; m-a, s-cn, 15 oct. 2009, leg. ab, det. mw. pilobolus crystallinus (f.h. wigg.) tode, on boar dung, opu, sośnia, ce-a close to edge of dune, 29 aug. 2013, leg. & det. mw. pilobolus nanus tiegh., on boar dung, opu, sośnia, ce-a close to edge of dune, 29 aug. 2013, leg. & det. mw. #piptocephalis lepidula (marchal) p. syd., on boar dung, opu, sośnia, ce-a close to edge of dune, 29 aug. 2013, leg. & det. mw. piptocephalis fimbriata m.j. richardson & leadbeater, on boar dung, opu, sośnia, ce-a close to edge of dune, 29 aug. 2013, leg. & det. mw. polysphondylium sp., soil, dilution method, kopu, biele suchowolskie fen, sp-c, 20 jun. 2008, leg. ab, det. mw; m-a, sp-c, 15 oct. 2009, leg. ab, det. mw. rhizopus arrhizus went & prins. geerl. [= rhizopus oryzae went & prins. geerl.], soil and plant debris, damp chamber method, opu, biały grąd, meadow, 27 aug. 2013, leg. & det. mw. rhizopus stolonifer (ehrenb.) vuill., plant debris, damp chamber method, bbnp buffer zone, osowiec village, carska droga, roaside, 30 aug. 2013, leg. & det. mw. rhopalomyces elegans corda, soil and plant debris, warcup method, kopu, biele suchowolskie fen, fdc, 20 jun. 2008, leg. & det. mw; s-cn, 15 oct. 2009, leg. & det. mw. sarocladium kiliense (grütz) summerb. [= acremonium kiliense grütz], (a), soil, warcup and dilution methods, kopu, biele suchowolskie fen, s-cn, 20 jun. 2008, leg. ab, det. mw; m-a, 15 oct. 2009, leg. ab, det. mw. sarocladium strictum (w. gams) summerb. [= acremonium strictum w. gams], (a), soil, warcup and dilution methods, kopu, biele suchowolskie fen, m-a, sp-c, 20 jun. 2008, leg. ab, det. mw; 27 aug. 2013 leg. & det. mw. 23 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park sordaria fimicola (roberge ex desm.) ces. & de not., (t), plant debris, kopu, biele suchowolskie fen, sp-c, 15 oct. 2009, leg. ab, det. mw. unidentified sporormiaceae, (t), soil, warcup method, kopu, biele suchowolskie fen, sp-c, 15 oct. 2009, leg. ab, det. mw. syncephalis nodosa tiegh., soil on mucoralean hyphae, kopu, biele suchowolskie fen, fdc, 15 oct. 2009, leg. ab, det. mw. syncephalis sphaerica tiegh., soil on mucoralean hyphae, kopu, biele suchowolskie fen, m-a, 20 jun. 2008, leg. ab, det. mw; m-a, sp-c, 15 oct. 2009, leg. ab, det. mw. syncephalis tenuis thaxt., soil, dump chamber, opu, biały grąd, meadow, 27 aug. 2013, leg. & det. mw. syzygites megalocarpus ehrenb., on humaria hemisphaerica fruitbody, opu, sośnia, mixed forest, 28 aug. 2013, leg. & det. mw. #talaromyces funiculosus (thom) samson, n. yilmaz, frisvad & seifert [= penicillium funiculosum thom.], (a), soil, dilution method, kopu, biele suchowolskie fen, fdc, m-a, s-cn, 20 jun. 2008, leg. ab, det. mw; sp-c, 15 oct. 2009, leg. ab, det. mw. #talaromyces rugulosus (thom) samson, n. yilmaz, frisvad & seifert [= penicillium rugulosum thom], (a), soil, dilution method, kopu, biele suchowolskie fen, m-a, sp-c, 20 jun. 2008, leg. ab, det. mw; m-a, s-cn, 15 oct. 2009, leg. ab, det. mw. trichocladium asperum harz, (a), soil, warcup and dilution methods, kopu, biele suchowolskie fen, sp-c, 20 jun. 2008 and 15 oct. 2009, leg. ab, det. mw. #trichoderma aggressivum samuels & w. gams, (a), on hoof fungus fomes fomentarius fruitbody, opu, biały grąd, solitary tree trunk on meadow, 27 aug. 2013, leg. & det. mw. trichoderma deliquescens (sopp) jaklitsch [= gliocladium viride matr.], (a), soil, dilution method, kopu, biele suchowolskie fen, m-a, 15 oct. 2009, leg. ab, det. mw. trichoderma hamatum (bonord.) bainier, (a), soil by dilution and warcup methods, kopu, biele suchowolskie fen, fdc, m-a, s-cn, sp-c, 20 jun. 2008, leg. ab, det. mw; sp-c, 15 oct. 2009, leg. ab, det. mw. trichoderma harzianum rifai, (a), soil, dilution and warcup methods, kopu, biele suchowolskie fen, fdc, s-cn, 20 jun. 2008, leg. ab, det. mw; s-cn, 15 oct. 2009, leg. ab, det. mw. trichoderma koningii oudem., (a), soil, dilution and warcup methods, kopu, biele suchowolskie fen, s-cn, 20 jun. 2008, leg. ab, det. mw; m-a, s-cn, 15 oct. 2009, leg. ab, det. mw. trichoderma polysporum (link) rifai, (a), soil, dilution and warcup methods, kopu, biele suchowolskie fen, fdc, sp-c, 20 jun. 2008, leg. ab, det. mw; m-a, sp-c, 15 oct. 2009, leg. ab, det. mw. trichoderma viride pers., (a), soil, dilution method, kopu, biele suchowolskie fen, fdc, m-a, s-cn, sp-c, 20 jun. 2008, leg. ab, det. mw; m-a, s-cn, sp-c, 15 oct. 2009, leg. ab, det. mw. verticillium sp., (a), soil, dilution method, kopu, biele suchowolskie fen, fdc, s-cn, sp-c, 20 jun. 2008, leg. ab, det. mw; m-a, 15 oct. 2009, leg. ab, det. mw. 24 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park discussion although the data presented in this paper are preliminary, the results of the second few-days-inventory carried out by pms, supplemented with limited data from the biele suchowolskie fen (two samples from each plant community) are promising for the future mycological studies in bbnp. the list of microfungal species is relatively extensive and includes taxa very rare or recorded for the first time in poland. in addition, about a half of the species from the current list was not observed during the previous pms survey in 2012 [1]. worth noting is also the ecological spectrum of the fungi presented in our paper, as the data on entomopathogens and soil fungi are very scarce in case of other mycologically studied polish national parks [50]. it concerns also most thoroughly studied białowieża national park [51] and tatra national park [52]. due to that lack of specific data and short-term research in bbnp the possibility to compare our results with other studies is greatly limited. the only practicable comparison concerns plant-associated species as this group of microfungi was extensively studied in nine of 23 polish national parks. the number of species recorded in bbnp in 2013 (190) accounts for more than one third of the number of species recorded in ojcowski, pieniński, and poleski national parks during several years’ studies using a route method (cf. [53]). surprisingly, current number of taxa from bbnp constitutes about a half of the number of species listed from long-term project crypto in the białowieża national park [51] and the projected jurassic national park [54] where permanent observation plots were used, but less than one third of taxa collected in the słowiński national park [55] using the same method. the currently reported number of plant-associated microfungi is however greater than that recorded using route method during systematic studies carried out in the lubelska upland and bug river valley (cf. [53]). the available data concerning arthropod associated fungi indicate that the number recorded in bbnp in 2013 (22 species) accounts for a little more than one third of the number of species recorded in białowieża np [56,57] and less than one quarter of taxa noted in wielkopolski national park [56]. the diversity of methods used for isolation and identification of soil fungi is another reason for limited comparison of the present results with literature data. for example, the methods used to study soil fungi vary strongly from one research to another. the most commonly used dilution method [58,59] was deeply criticized by warcup [60]. his soil-plate method is still widely used in a direct way or with modifications [61–63]. since the beginning of twenty-first century new method of a cultivation-independent analysis and direct dna extraction and sequencing were implemented [64,65] to study soil fungal communities. the results of traditional and molecular studies are incomparable, and the lists of microfungal species found in soil are difficult to interpret. the dilution method favors fungi producing phialospores while fungi that are not sporulating readily on agar media could be detected by direct sequencing [64]. yeasts such as trichosporon spp. are identified in soils more often in molecular studies than when using other techniques [66–68]. some quite rare, specialized fungi could be also isolated from soil using trap method [69]. these fungi often remain unidentified in molecular studies due to the lack of reference sequences. the majority of investigations of soil fungal communities fall into one of three categories: systematic studies, biochemical research and phytopathology. most of them is focused on plant rhizosphere – a microecological zone in direct proximity of plant roots. symbiotic and pathogenic fungi could significantly affect the yield, and studies are restricted to economically important plants. the general ecological investigations of soil fungi are still very rare [70] and assignment of any taxa to specific habitats is impossible yet. our present studies are preliminary and should be supplemented with taxa identified by pyrosequencing. the studies by buée and collaborators [65] and lim and collaborators [71] indicate that the richness of fungi in soils is much bigger than expected. finally, one should note that only holistic approach could lead us to understanding of the specificity of soil mycocoenoses. 25 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1067 ruszkiewicz-michalska et al. / micromycetes in biebrza national park key outcomes ■ nineteen taxa were found for the first time in poland: candelabrum spinulosum, cephalotrichum nanum, cercosporella lindaviana, colletotrichum veratrina, daldinia decipiens, discosia fitzpatrickii, d. kabati, d. potentillae, d. pulmonariae, d. quercicola, helicocephalum sp., nemania aenea, mycosphaerella osborniae, phaeosphaeria inclusa, phomopsis amygdaliana, pilobolus nanus, piptocephalis fimbriata, seimatosporium foliicola, and sphaceloma viburni. ■ thirty-one taxa that may be considered as rare, including plant-inhabiting fungi (ascochyta equiseti, asteromella pruni-mahaleb, discosia minuta, fusicladium betulae, hypomyces rosellus, helminthosphaeria clavariarum, leveillula helichrysi, monilinia padi, peronospora agrimoniae, phyllosticta pyrolae, septoria jasiones, septoria tabacina, stephanoma strigosum), and four isolated from soil (talaromyces funiculosus, talaromyces rugulosus, piptocephalis lepidula, trichoderma aggressivum). ■ 165 species are reported as new to the area of the biebrza national park. although some taxa are recorded in poland for the first time, one should note that the knowledge about occurrence and distribution of microfungi without economical impact is still very limited. thus all final conclusions basing on available data should be treated with caution. acknowledgments we are deeply grateful to mr. roman skąpski (director of bbnp) as well as to ms. agnieszka henel, mr. cezary werpachowski and mr. krzysztof frąckiel, and other employees of the biebrza national park for collecting fungi, offering their facilities, helping in the implementation of our research and creating the very friendly atmosphere during our visit. we are grateful to the following persons for permission to use their materials in our paper: abiba boulahdjel (university of warsaw), grażyna domian (regional directorate for environmental protection in szczecin), błażej gierczyk (adam mickiewicz university in poznań), tomasz leski and marcin pietras (institute of dendrology, polish academy of sciences, kórnik), małgorzata stasińska (university of szczecin), and natalia michalska (łódź). we thank also błażej gierczyk and anna kujawa (institute for agricultural and forest environment) for identification of helminthosphaeria clavariarum and hypomyces rosellus and jeremi kołodziejek (university of łódź) for 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1995. 70. badurowa m, badura l, further investigations on the relationships between soil fungi and macroflora. acta soc bot pol. 1967;36(3):515–529. http://dx.doi.org/10.5586/ asbp.1967.049 71. lim yw, kim bk, kim c, jung hs, kim bs, lee jh, et al. assessment of soil fungal communities using pyrosequencing. j microbiol. 2010;48(3):284–289. http://dx.doi. org/10.1007/s12275-010-9369-5 http://dx.doi.org/10.1007/bf02052135 http://dx.doi.org/10.1016/b978-0-12-395699-6.50007-9 http://dx.doi.org/10.1097/00010694-196108000-00001 http://dx.doi.org/10.1128/aem.69.7.3758-3766.2003 http://dx.doi.org/10.1111/j.1469-8137.2009.03003.x http://dx.doi.org/10.1139/w99-055 http://dx.doi.org/10.1139/w99-055 http://dx.doi.org/10.1016/s1567-1356(00)00002-7 http://dx.doi.org/10.1016/s1567-1356(00)00002-7 http://dx.doi.org/10.1099/ijs.0.02859-0 http://dx.doi.org/10.5586/asbp.1967.049 http://dx.doi.org/10.5586/asbp.1967.049 http://dx.doi.org/10.1007/s12275-010-9369-5 http://dx.doi.org/10.1007/s12275-010-9369-5 abstract introduction material and methods results plant-associated fungi fungi associated with arthropods fungi and dictyostelia isolated from soil and associated with plant debris and excrements discussion key outcomes acknowledgments references 2016-01-29t11:40:19+0000 piotr otręba the influence of tin ions on growth and enzymatic activity of entomopathogenic fungi 1 of 10published by polish botanical society acta mycologica original research paper the influence of tin ions on growth and enzymatic activity of entomopathogenic fungi łukasz łopusiewicz1*, kinga mazurkiewicz-zapałowicz2, marta koniuszek2, cezary tkaczuk3, artur bartkowiak1 1 center of bioimmobilization and innovative packaging materials, west pomeranian university of technology in szczecin, klemensa janickiego 35, 71-270 szczecin, poland 2 department of hydrobiology, ichthyology and biotechnology of reproduction, west pomeranian university of technology in szczecin, kazimierza królewicza 4, 71-550 szczecin, poland 3 department of plant protection and breeding, siedlce university of natural sciences and humanities, bolesława prusa 14, 08-110 siedlce, poland * corresponding author. email: lukasz.lopusiewicz@zut.edu.pl abstract in this in vitro study, the influence of tin ions at concentrations of 1–1,000 ppm on the development and enzymatic activity of four entomopathogenic fungi (beauveria bassiana, b. brongniartii, isaria fumosorosea, and metarhizium robertsii), that are commonly used in biological plant protection, are examined. each of the fungal species tested reacted differently to contact with the sn2+ ions at the tested concentrations. exposure to sn2+ ions affected the rate of development, morphology, and enzymatic activity of fungi. of the four fungal species studied, m. robertsii was the most resistant and showed complete growth inhibition at the highest sn2+ concentration tested (1,000 ppm). for the other entomopathogenic fungi, the fungicidal effect of sn2+ ions was noted at the concentration of 750 ppm. exposure to sn2+ ions (up to 500 ppm) resulted in enhanced biochemical activity; and all entomopathogens that were tested showed increased production of n-acetyl-β-glucosaminidase (nag) as well as several proteases. moreover, b. brongniartii and m. roberstii showed increased lipases synthesis. these changes may increase the pathogenicity of the fungi, thereby making them more effective in limiting the population of pest insects. the exposure of the entomopathogenic fungi to a medium containing sn2+ ions, at concentrations that were appropriate for each species, induced hyperproduction of hydrolases, which might be involved in aiding the survival of entomopathogenic fungi in the presence of heavy metals. this study shows that the fungistatic effect of sn2+ on entomopathogenic fungi did not restrict their pathogenicity, as evidenced by the stimulation of the production of enzymes that are involved in the infection of insects. keywords heavy metals; beauveria; isaria; metarhizium; microfungi introduction metals are an integral part of all ecosystems and occur in both elemental and ore forms throughout nature. industrialization and urbanization, in this and preceding centuries, have generated a tremendous amount of soil, water, and air pollution, which interferes with homeostasis in the ecosphere [1]. the utilization of pesticides, chemical fertilizers, and preservatives, particularly in agriculture, contributes to fluctuations in the chemical doi: 10.5586/am.1127 publication history received: 2019-04-04 accepted: 2019-06-04 published: 2019-12-19 handling editor wojciech pusz, faculty of life sciences and technology, wrocław university of environmental and life sciences, poland authors’ contributions łł, kmz, and mk: designing and running the experiment, analyzing the data; ct: isolation of entomopathogenic fungi; ab: analyzing the data; all authors contributed to the manuscript writing funding publishing of the manuscript was covered by the statutory funds of the faculty of food sciences and fisheries, west pomeranian university of technology szczecin. competing interests no competing interests have been declared. copyright notice © the author(s) 2019. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation łopusiewicz ł, mazurkiewiczzapałowicz k, koniuszek m, tkaczuk c, bartkowiak a. the influence of tin ions on growth and enzymatic activity of entomopathogenic fungi. acta mycol. 2019;54(2):1127. https:// doi.org/10.5586/am.1127 mailto:lukasz.lopusiewicz%40zut.edu.pl?subject=the%20influence%20of%20tin%20ions%20on%20growth%20and%20enzymatic%20activity%20of%20entomopathogenic%20fungi https://doi.org/10.5586/am.1127 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1127 https://doi.org/10.5586/am.1127 2 of 10© the author(s) 2019 published by polish botanical society acta mycol 54(2):1127 łopusiewicz et al. / the influence of tin on entomopathogenic fungi composition of the ecosphere and to the disturbance of interactions between organisms in the soil [2]. life has evolved in environments that are rich in various metals and all cells have incorporated metal ions into their essential cellular functions [3]. consequently, life forms that are continuously exposed to potentially toxic conditions have evolved mechanisms of metal homeostasis and metal resistance to adapt to the metals that are present in their environments. this requires mechanisms that ensure sensitivity towards different metal species at the concentration at which they are present in the environment [3]. the introduction of heavy metal compounds into the environment generally induces morphological and physiological changes in microbial communities [4]. it is well established that heavy metals interfere with the physiological, enzymatic, and reproductive processes of organisms, thereby affecting the ecosystem. entomopathogenic fungi (epf) are a group of highly specialized microorganisms that can infect arthropods, including insects that are pests of crop plants [5]. the ability of epf to infect insect pests makes this group of fungi particularly important in biological plant protection [6]. epf occur primarily in the soil and constitute an essential part of the organic biomass [7,8]. soil can provide a substrate for the maintenance of a natural reservoir of many epf. for this reason, soil can be inoculated with epf by an infected insect entering the soil, by the deposition of spores on the surface of the soil or by natural dispersion mechanisms [9]. the presence and pathogenicity of many epf depend on their interactions with host organisms, prevailing climatic conditions, as well as other biotic and abiotic factors, which include contact with heavy metals. laboratory studies have shown that heavy metals influence growth, metabolism, and pathogenicity of epf [2,6,10–12]. tin (sn) is a naturally occurring heavy metal that is present at an average concentration of 2 mg/kg in the earth’s crust. the concentration of sn in the environment is, however, highly variable and is dependent both on the use of sn and the release of sn from sn-containing entities. the release of sn can occur by natural means such as the weathering of rocks or volcanic eruptions or due to anthropogenic activities, such as industrial processes, agriculture, and mining [13]. normal concentrations of sn in unpolluted soils range from <1 mg/kg to 200 mg/kg; the sn present in the soil occurs in two oxidative states (ii and iv). in the soil, sn usually has limited mobility and is usually tightly bound in the top soil [14]. however, due to the increase in anthropogenic activities such as agriculture, which release sn products into the environment, concentrations of sn may be elevated in certain areas [14]. sn can combine with chemicals like chlorine, sulfur, or oxygen to form inorganic sn compounds (i.e., chlorides, sulfides, and oxides) [15]. inorganic sn compounds are used as pigments in the ceramic and textile industry. tin (ii) chloride, sncl2, is used as a protective tinplate coating for steel sheet for use in manufacturing, processing, and packaging of foods as well as in biocidal preparations. sn is used in canned foods to protect the steel base from corrosion both externally, due to aerobic conditions, and internally, when the sn is under anaerobic conditions and in contact with food. the sn in canned food is likely to be in the inorganic salt form as opposed to the covalently-bound organometallic forms. the corrosion of cans may be one of ways in which sn is released into the environment as pollution. although the biological functions of sn have not been described to date, it is difficult to agree with the opinion that sn is a nonessential metal that is of no importance to organisms [15,16]. sn compounds affect the physiology of bacteria and fungi [17–21], plants [13,14], and animals [15,17,22,23]. interactions of sn with microorganisms are ambiguous, as although sn and its compounds can be metabolized by some microorganisms, they are toxic to others. microbial interactions with sn are important, because microbes are at the base of many food webs and are likely to be significant in the environmental transformation of sn compounds. this suggests that microbes may have significant potential in the remediation of sn-polluted waste streams and of sn-polluted ecosystems [24]. little is known at this time about the effect of sn on the growth and biochemical activity of epf. this study aims to determine the sensitivity of four epf (beauveria bassiana, b. brongniartii, isaria fumosorosea, and metarhizium robertsii) to increasing concentrations of sn2+. in addition, this study aims to understand the response of the epf to sn, which determines their potential of epf in biological pest control. 3 of 10© the author(s) 2019 published by polish botanical society acta mycol 54(2):1127 łopusiewicz et al. / the influence of tin on entomopathogenic fungi material and methods fungal strains beauveria bassiana (bals.-criv.) vuill. (uph34), b. brongniartii (sacc.) petch (uph42), isaria fumosorosea (wize) kepler, b. shrestha & spatafora (uph62), and metarhizium robertsii j. f. bisch., s. a. rehner & humber (wa27856) fungal strains were obtained from the fungal collection at the department of plant protection and breeding, siedlce university of natural sciences and humanities (siedlce, poland). the strains were isolated near siedlce (masovian province, poland) from the soil of arable fields by using the galleria bait method [25]. before the experiments, all isolates were grown on a sabouraud medium (biocorp, poland) and stored at 4°c. media and the preparation of epf inocula the influence of metal on epf was tested on solid pda medium (biocorp, poland) supplemented with sn (ii) chloride (sigma-aldrich, germany). sncl2·2h2o salt was added to pda medium after autoclaving (when the temperature of the medium reached approx. 50°c) to achieve concentrations of 1, 10, 50, 100, 250, 500, 750, and 1,000 ppm of sn2+ ions. the medium was then placed on a magnetic stirrer and when it cooled, it was poured into 90-mm petri dishes. the pda medium, which lacked added sn2+ ions served as the control medium. inocula were prepared from 10-day old fungal colonies grown on pure pda medium. aerial hyphae of epf strains were collected and a suspension of fungal spores in sterile physiological saline was prepared. the concentration of fungal spores was calculated using a thoma counting chamber and was approx. 1.0 × 109 ml−1. twenty-μl drops of epf suspensions were transferred using an automatic pipette to the center of the test plates. five biological repeats were prepared for each sn concentration as well as for the control, and the plates were incubated at 25°c for 18 days. growth response studies and the determination of the minimum inhibitory concentration of sn the development of fungi was evaluated using the tolerance index (ti) as previously described [26]. to compare the ti of the epf strains, the radius of the colony extension on pda medium supplemented with sn2+ ions at different concentrations was measured against the control medium (pda devoid of added sn2+ ions). the radial growth was evaluated by performing four measurements in millimeters, each measurement passed through the center of inoculated epf material. the minimum inhibitory concentration (mic) was defined as the minimum inhibitory concentration of heavy metal in the medium that inhibited the visible growth of tested epf strains. if no fungal growth was observed after the incubation period, that sn2+ ion concentration was considered the highest metal concentration tolerated by the epf tested. the determination of fungal enzymatic activity the api-zym test (biomérieux, lyon, france) was used to semiquantitatively determine the activity of 19 hydrolytic fungal enzymes including alkaline phosphatase (2), esterase (c4) (3), esterase lipase (c8) (4), lipase (c14) (5), leucine arylamidase (6), valine arylamidase (7), cystine arylamidase (8), trypsin (9), chymotrypsin (10), acid phosphatase (11), naphthol-as-bi-phosphohydrolase (12), α-galactosidase (13), β-galactosidase (14), β-glucuronidase (15), α-glucosidase (16), β-glucosidase (17), n-acetyl-β-glucosoaminidase (nag; 18), α-mannosidase (19), and α-fucosidase (20) according to the manufacturer’s instructions. fungal cultures that were grown on pda without and with sn2+ ions at concentrations of 1, 100, and 500 ppm for 14 days were transferred into a sterile physiological saline solution. the api-zym strips were inoculated with the resuspended epf culture and then incubated at 37°c for 4 h. hydrolytic 4 of 10© the author(s) 2019 published by polish botanical society acta mycol 54(2):1127 łopusiewicz et al. / the influence of tin on entomopathogenic fungi activity was determined in nanomoles (nm) of hydrolyzed substrate in a 5-grade color scale, ranging from 0 to 5, as described by manufacturer. zero indicates a negative reaction with no enzyme production, 1 indicates 5 nm hydrolyzed substrate, 2 indicates 10 nm hydrolyzed substrate, 3 indicates 20 nm hydrolyzed substrate, 4 indicates 30 nm hydrolyzed substrate, and 5 indicates 40 nm or more of hydrolyzed substrate. results compared to the control, all epf species tested showed a delay in linear growth on pda medium supplemented with sn2+ ions at concentrations of 1–500 ppm (fig. 1–fig. 4). the complete inhibition of b. bassiana, b. brongniartii, and i. fumosorosea development occurred at an mic of 750 ppm, whereas inhibition of m. robertsii development occurred at an mic 1,000 ppm. exposure to sn also resulted in morphological changes in the epf mycelia. at a concentration of 250 and 500 ppm, there was a reduction in the aerial hyphae of m. roberstii. at sn2+ concentrations higher than 500 ppm, the aerial hyphae were no longer visible for b. bassiana and b. brongniartii. there were no morphological changes in the mycelia of i. fumosorosea regardless of the sn2+ concentration present in the growth medium, the hyphae were indistinguishable from those that developed under control conditions. the exposure of b. bassiana colonies to pda medium containing sn2+ concentrations between 1–50 ppm resulted in the formation of a white halo around the fungal colonies. in contrast, a pink pigment and a dark halo was produced around b. brongniartii colonies on pda plates supplemented with 1 and 10 ppm of sn2+. no color reactions were observed around i. fumosorosea and m. robertsii colonies. the changes in the concentration of sn2+ ions resulted in changes in the enzymatic activity of the epf tested (tab. 1). when compared to the enzymatic activity on control media, the growth of b. bassiana in the presence of sn2+ at a concentration of 1 ppm did not result in the inhibition of synthesis of any enzyme. in contrast, increased production of leucine arylamidase (6), valine arylamidase (7), acid phosphatase (11), α-galactosidase (13) and nag (18) was detected. at a concentration of 500 ppm sn2+, increased activity of acid phosphatase (11), naphthol-as-bi phosphohydrolase (12), and β-glucosidase (17) was detected when compared to the control b. bassiana sample. however, compared to the activity detected at 1 ppm sn2+, decreased leucine arylamidase (6) and nag activities (18) were detected. the growth of b. brongniartii in medium containing 1 ppm of sn2+ ions resulted in increased alkaline phosphatase (2), lipase esterase (c8) (4), acid phosphatase (11), α-galactosidase (13), β-galactosidase (14), α-glucosidase (16), β-glucosidase (17), nag (18), and α-mannosidase (19) activity but reduced esterase (c4) (3) and cystine arylamidase (8) activity compared to those of the control sample. growth in medium containing 100 and 500 ppm sn2+ ions resulted in increased synthesis of leucine arylamidase (6), valine arylamidase (7), acid phosphatase (11), naphthol-as-bi-phosphohydrolase (12), β-galactosidase (14), β-glucosidase (17), and nag (18). growth of i. fumosorosea in medium containing a concentration of 1 ppm sn2+ resulted in a reduction in naphthol-as-bi-phosphohydrolase (12), β-galactosidase (14), and β-glucosidase levels compared to those in the control. the other enzymes were produced at the same levels in the control and sn2+ containing medium. the development of i. fumosorosea in medium containing sn2+ at 100 and 500 ppm resulted in increased levels of cystine arylamidase (8), naphthol-as-bi phosphohydrolase (12), nag (18), and α-fucosidase (20) compared to those in the control. when compared to the control sample, the growth of m. roberstii on media containing different concentrations of sn2+ ions resulted in increased activity of the majority of the tested enzymes. when exposed to media containing 1 and 100 ppm sn2+ ions, no enzyme was inhibited; however, the levels of acid phosphatase (11) and naphtholas-bi phosphohydrolase (12) and nag (18) were elevated compared to those in the control. the synthesis of these enzymes was limited when the medium contained sn2+ ions at a concentration of 500 ppm; however, the levels remained higher than those found in the control. 5 of 10© the author(s) 2019 published by polish botanical society acta mycol 54(2):1127 łopusiewicz et al. / the influence of tin on entomopathogenic fungi 0.5 0.55 0.6 0.65 0.7 0.75 0.8 0.85 0.9 0.95 1 1.05 1.1 1.15 3 6 9 12 15 18 to le ra nc e in de x days beauveria bassiana 1 ppm 10 ppm 50 ppm 100 ppm 250 ppm 500 ppm 0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45 0.5 0.55 0.6 0.65 0.7 0.75 0.8 0.85 0.9 0.95 1 1.05 1.1 1.15 1.2 3 6 9 12 15 18 to le ra nc e in de x days beauveria brongniartii 1 ppm 10 ppm 50 ppm 100 ppm 250 ppm 500 ppm 0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45 0.5 0.55 0.6 0.65 0.7 0.75 0.8 0.85 0.9 0.95 1 1.05 1.1 1.15 3 6 9 12 15 18 to le ra nc e in de x days isaria fumosorosea 1 ppm 10 ppm 50 ppm 100 ppm 250 ppm 500 ppm fig. 1 effect of an increasing sn2+ concentration on the tolerance index of beauveria bassiana over an 18-day incubation period. fig. 2 effect of an increasing sn2+ concentration on the tolerance index of beauveria brongniartii over an 18-day incubation period. fig. 3 effect of an increasing sn2+ concentration on the tolerance index of isaria fumosorosea over an 18-day incubation period. 6 of 10© the author(s) 2019 published by polish botanical society acta mycol 54(2):1127 łopusiewicz et al. / the influence of tin on entomopathogenic fungi discussion there are few studies that have focused on the toxicity of inorganic sn compounds towards microorganisms. this may be due to the widespread belief that inorganic sn species hydrolyze to form insoluble and nontoxic sn oxides or sn hydroxides. most of the studies that have studied the toxicity of sn have concentrated on organotin compounds [27]. the mechanisms by which inorganic sn exerts its toxic effects, and the influence of these compounds on fungal physiology, is unclear. it is possible that it may be complex to understand. tobin and cooney [17] observed that the inorganic sn ions (sn2+ and sn4+) bind to candida maltosa yeast cells at levels of 0.3 and 0.23 mm sn/g 0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45 0.5 0.55 0.6 0.65 0.7 0.75 0.8 0.85 0.9 0.95 1 1.05 1.1 3 6 9 12 15 18 to le ra nc e in de x days metarhizium robertsii 1 ppm 10 ppm 50 ppm 100 ppm 250 ppm 500 ppm 750 ppm fig. 4 effect of an increasing sn2+ concentration on the tolerance index of metarhizium robertsii over an 18-day incubation period. tab. 1 the enzymatic activity of entomopathogenic fungi. enzyme activity was determined with the api-zym test. enzyme b. bassiana b. brongniartii i. fumosorosea m. robertsii c sn1 sn2 sn3 c sn1 sn2 sn3 c sn1 sn2 sn3 c sn1 sn2 sn3 1 alkaline phosphatase 2 1 1 1 3 4 3 3 1 1 1 1 1 2 2 1 2 esterase (c4) 1 1 0 2 3 1 3 3 1 1 1 1 1 2 2 1 3 esterase lipase (c8) 2 1 1 1 1 3 1 1 1 1 1 1 1 2 1 1 4 lipase (c14) 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 5 leucine arylamidase 2 3 3 1 3 3 4 4 1 1 1 1 1 3 3 1 6 valine arylamidase 1 1 1 1 2 2 3 3 1 1 1 1 1 3 3 1 7 cystine arylamidase 1 1 1 1 2 1 2 2 1 1 2 2 1 1 1 1 8 trypsin 1 1 1 1 1 1 2 2 1 1 1 1 1 2 1 1 9 chymotrypsin 1 1 1 1 1 1 2 2 1 1 1 1 1 2 1 1 10 acid phosphatase 1 1 3 2 3 4 4 4 2 2 2 1 1 5 5 1 11 naphthol-as-bi-phospohydrolase 2 3 1 2 4 4 5 5 3 1 4 4 1 5 5 1 12 α-galactosidase 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 4 13 β-galactosidase 2 2 1 1 3 4 5 5 2 1 2 2 1 2 2 3 14 β-glucuronidase 1 2 1 1 1 1 1 1 1 1 1 1 1 1 1 3 15 α-glucosidase 1 1 1 1 1 2 1 1 1 1 1 1 1 1 1 1 16 β-glucosidase 1 3 3 1 1 2 3 3 2 1 1 1 3 4 4 3 17 n-acetyl-β-glucosoaminidase 2 3 2 1 2 4 4 4 1 1 4 4 4 5 5 3 18 α-mannosidase 1 2 2 1 2 3 4 4 1 1 1 1 1 2 2 1 19 α-fucosidase 0 0 0 0 1 2 4 4 1 1 2 2 1 2 3 1 c – control; sn1 – sn concentration 1 ppm; sn2 – sn concentration 100 ppm; sn3 – sn concentration 500 ppm. 7 of 10© the author(s) 2019 published by polish botanical society acta mycol 54(2):1127 łopusiewicz et al. / the influence of tin on entomopathogenic fungi cells, respectively however sn did not inhibit growth of the c. maltosa at concentrations up to 0.8 mm. the inorganic sn did not cause the leakage of potassium from the yeast cells however organotins did affect the physiological state of yeast. with reference to basidiomycota, kähkönen et al. [28] showed that although there are individual species of fungi with a high tolerance towards inorganic sn compounds (sncl4·5h2o), there are many more species that lack this tolerance. it has been reported that sncl2 is capable of promoting the formation of reactive oxygen species (ros), which are responsible for the oxidative stress that causes dna damage [29]. according to dantas et al. [30], the sn2+ toxicity mechanism may be related to fenton-like reactions. ros, such as the hydroxyl radical (•oh), which is produced in the cells, is capable of damaging important cellular targets, including membranes and dna. metal ions, including a number of transition metals such as iron, copper, zinc, and chromium, are able to mediate fenton or fenton-like reactions that generate ros in the presence of hydrogen peroxide (h2o2) [30]. at higher concentrations, some of the metal ions, and particularly the heavy metals, interfere negatively with cellular metabolism as they may inactivate proteins and damage dna [31]. the genotoxic potential of sn may also be significantly modulated by other non-dna repair or membrane transport-related physiological parameters. this includes the quality and quantity of enzymatic and nonenzymatic scavengers of metal-induced ros, which may be a crucial factor that influences the physiological response of epf. inorganic sn increases the frequency of chromosomal aberrations, sister chromatid exchange and reduces cell proliferation [15,32]. inorganic sn also induces rapid and prolonged suppression of dna synthesis resulting in changes in gene expression as reported by mclean et al. [33]; these authors also noticed that sncl2 produced single-strand breaks in dna. the toxicity of inorganic sn (sncl2) has also been demonstrated towards microorganisms that live in saline estuaries. hallas et al. [27] suggest that this may be due to the interaction of the metal with polysaccharides and the consequent formation of cytotoxic complexes. this study also demonstrated the toxicity of sn2+ ions to microorganisms. this finding is confirmed by the individual reactions of epf to sn that were observed in this study. epf exhibited differential tolerance to the presence of sn, there were visible differences in development as well as differences in epf biochemical activity; these demonstrate that sn exerted a degree of toxicity towards microorganisms. the morphological changes and the colorful reactions that occur when epf are in contact with sn2+ ions may be the result of physiological disturbances; this has been shown in other microorganisms [26,34,35]. the production of organic acid-based pigments (such as salts of oxalic acid and citric acid), in fungal cells or their secretion into the environment could be harnessed to precipitate metal ions; this could be used in mechanical detoxification. there are two mechanisms that have been proposed to explain the tolerance of fungi to heavy metals. these include the extracellular sequestration of the metal ions by chelation or cell wall binding and the intracellular and physical sequestration of metals by binding them to proteins or other ligands to prevent the metal ion from damaging the metal sensitive cellular targets [34,35]. among the tested epf, the highest tolerance to contact with sn2+ was demonstrated by m. robertsii, with an mic value of 1,000 ppm. for the other epf species, the mic values were 750 ppm. the fungistatic effect of the sn2+ ions is thought to be due to the inhibition of the logarithmic phase of fungal growth, specifically the trophophase, which is usually dependent on environmental conditions [1,26,34,36]. the consequence of the sn-dependent physiological changes is the delay in the stationary phase, the idiophase, which is important for fungi because it is associated with the production of key secondary metabolites, such as mycotoxins [37]. limiting the toxigenic potential of epf may reduce their pathogenicity to insects. insect mycoses are, however, also dependent on other factors that interact with or are independent of mycotoxins. in addition to their mycotoxin-forming ability, the effectiveness of entomopathogens in the reduction of insect populations is also determined by the activity of their enzymes, especially those that are involved in the degradation of the epicuticle, which is the outer body of the insect. these enzymes include those that are active in the digestion of chitin, which constitutes 60% of the dry matter of insects’ epicuticle. chitin is composed of polymers of n-acetyl-d-glucosamine whose structure is destroyed by the enzyme nag. the present study indicates an interesting and as yet unreported effect of sn; stimulation with sn resulted in the elevated 8 of 10© the author(s) 2019 published by polish botanical society acta mycol 54(2):1127 łopusiewicz et al. / the influence of tin on entomopathogenic fungi production of this enzyme by b. bassiana (at 1 ppm), b. brongniartii (at 1, 100, and 500 ppm), and m. robertsii (at 1 and 100 ppm). overproduction of nag was also demonstrated in i. fumosorosea at concentrations of 100 and 500 ppm. while this may significantly accelerate the infection and development of insect mycoses, this response may also represent the manifestation of the defense mechanism of these microorganisms. according to pusztahelyi et al. [38], nag is a high molecular-weight hydrolytic lysosomal enzyme, which breaks chemical bonds of glycosides and amino sugars that form structural components in many tissues. nag is necessary for the degradation and disposal of various parts of the cell, including the cell membranes. the degradation of insect cuticles may also be accelerated by elevated lipases activity. lipases, in addition to increasing the degree of adhesion of fungal spores to insect cuticles, result in the hydrolysis of lipid compounds and phosphate esters, which leads to disturbances in the permeability of biological membranes [39,40]. hence, the increased lipase and esterase activities demonstrated in b. brongniartii and m. robertsii that are in contact with sn2+ ions at 1 ppm in this study may have an important and practical application. this aspect of epf activity also permits the involvement of the proteases in the disease process in insects. there are numerous reports that show that proteolytic activity determines the pathogenicity of epf [41,42]. epf proteases appear to be less sensitive to contact with sn2+ ions, as evidenced by their increased production by b. bassiana, b. brongniartii, and m. robertsii at sn2+ ion concentrations between 1–500 ppm and by i. fumosorosea at sn2+ ion concentrations of 100 and 500 ppm. it is tempting to speculate that the activation of proteases may permit the increased hydrolysis of insect cuticle proteins even at toxic levels of sn2+ ions. the amino acids released by this process may be used by the epf as nutrients essential for their development [41,42]. in this context, the overproduction of hydrolases by epf may have an ambiguous effect. on the one hand, this reaction may be considered a factor that leads to the increased pathogenicity of the insect pathogens. on the other hand, this may lead to an increase in mycelial survival in toxic conditions, in this case when the mycelia are in contact with sn2+ ions. the diverse and ambiguous reactions of the epf to the presence of inorganic sn in the present study indicates the importance of broadening this study to other microorganisms. microorganism contact with inorganic sn2+ ions in the environment is toxic but it can also lead to reactions in the microorganisms that could strengthen their beneficial role in the ecosystem. in conclusion, in this report epf that were tested exhibited a sensitivity to sn2+ ions. the presence of these ions modified the development of the epf and their biochemical activity. sn2+ ions have fungistatic activity and could be used to restrict their growth in the environment as well as to influence the fungal communities in contaminated soils. stimulation of the synthesis of extracellular enzymes, including nag as well as some proteases and lipases, due to the action of sn2+ ions translates directly into strengthening the role of these microorganisms in their pathogenesis of insects, and so the effectiveness of causing mycoses in plant pests. this study suggests that the use of epf in biological plant protection in practice may differ from that detected in controlled conditions; 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https://doi.org/10.1111/j.1574-6968.2002.tb11314.x https://doi.org/10.4236/aer.2014.22007 abstract introduction material and methods fungal strains media and the preparation of epf inocula growth response studies and the determination of the minimum inhibitory concentration of sn the determination of fungal enzymatic activity results discussion references notes on the distribution of lichen biota of podlasie. iii. nowosady village, podlaskie province (north-eastern poland) 1 of 5published by polish botanical society acta mycologica original research paper notes on the distribution of lichen biota of podlasie. iii. nowosady village, podlaskie province (north-eastern poland) sylwia kiercul* department of environmental protection and management, białystok university of technology, wiejska 45a, 15-351 białystok, poland * email: sylwiakiercul@op.pl abstract the present study was undertaken to evaluate the biodiversity of lichen species in nowosady village and surrounding areas. this work was conducted in 2014 (in august) and biodiversity of lichen species growing on tree bark and bushes, on dead wood (anthropogenic origin), glacial erratics, concrete, mortared walls and other specific substrates like eternit roof slates has been assessed. the lichen species represented morphologically diverse forms: crustose (38%), foliose (38%), fruticose (13%), dimorphous (5%), placodial (3%) and squamulose (3%). they belonged to different ecological types including epiphytes (27 species), epixyles (18) and epilithes (12). out of 39 species identified in nowosady village, five are included in the polish red list of lichens: bryoria fuscescens, evernia prunastri, hypogymnia tubulosa, ramalina farinacea and ramalina fraxinea. four taxons from the study area are under statutory protection of species. one species, ramalina fraxinea is under full protection and 3 species (bryoria fuscescens, evernia prunastri and hypogymnia tubulosa) are under partial protection. keywords lichens; distribution; nowosady village; podlasie; ne poland this issue of acta mycologica is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. introduction nowosady is a small village located at the border between two mesoregions: białostocka upland and narew river upper valley. the lichen biota of these geographical areas is poorly known. so far, the diversity of lichen biota of several polish cities has been investigated, including gdańsk, sopot, gdynia [1], olsztyn [2] and świdnik [3]. the lichen species inhabiting smaller towns on podlasie, like drohiczyn [4], mielnik [5], boćki [6] and białowieża [7] have been also described. the diversity of lichen biota of orchards in different towns has been studied in slovakia, italy and poland [8], and in several polish villages or rural areas, including warmia plain [9], klewinowo [10] and hermanówka [11]. the main goal of the present study was to evaluate the biodiversity of species in the nowosady village and its surrounding areas, taking into account their natural environmental conditions. doi: 10.5586/am.1053 publication history received: 2015-02-21 accepted: 2015-06-24 published: 2015-08-05 handling editor maria rudawska, institute of dendrology of the polish academy of sciences, poland funding the study was supported by the polish ministry of science and higher education, project mb/ wbiiś/3/2014. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation kiercul s. notes on the distribution of lichen biota of podlasie. iii. nowosady village, podlaskie province (northeastern poland). acta mycol. 2015;50(1):1053. http://dx.doi. org/10.5586/am.1053 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:sylwiakiercul%40op.pl?subject=notes%20on%20the%20distribution%20of%20lichen%20biota%20of%20podlasie.%20iii.%20nowosady%20village%2c%20podlaskie%20province%20%28north-eastern%20poland%29 http://dx.doi.org/10.5586/am.1053 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1053 http://dx.doi.org/10.5586/am.1053 2 of 5© the author(s) 2015 published by polish botanical society acta mycol 50(1):1053 kiercul / species richness of lichens in the nowosady village study area nowosady is a small village in podlaskie province located on północnopodlaska lowland, at the border of two mezoregions: narew upper valley and białostocka upland [12], in the cg-square of the atpol grid square system [13]. the village occupies a hilly upland area with elevations reaching 157.3 m above sea level. the terrain was formed by last middle-poland glaciation. geologically, the study area was formed by the ground moraine. the cover of postglacial deposits reaches 200 m and is composed of mainly sands and gravels. the nowosady village lies in the area where the influence of the moderate warm and moderate humid continental climates meet. the overlapping of the climates and the relief of this area have considerable effect on the flora of the village studied. the agricultural landscape of this area is intertwined by the fragments of mixed forests, comprising fresh forests (peucedano-pinetum) and dry forests (cladonio-pinetum), sometimes with communities comprising birch or saplings of oaks and herbal layer dominated by field weed species. the age of these forests is estimated to be 20–40 and 40–60 years. material and methods the study was carried out in nowosady village and in the nearby areas, in august 2014. the lichen biota was analyzed using methods that are generally accepted in lichenology. a result of the field study was the compilation of a floristic list of all lichen species. materials for the herbarium, deposited at the herbarium of the institute of biology, university of białystok, were identified on the basis of morphological traits of their thalli, anatomical details and the thallus content of specific substances reacting with chemical reagents inducing color changes. the nomenclature for lichen species was adopted from diederich et al. [14]; for melanohalea and melanelixia genera from blanco et al. [15]. species included in the “red list of the lichens in poland” were quoted after cieśliński et al. [16] and protected species after the regulation of the minister of the environment dated october 9, 2014, on the protection of species of fungi (oj from 2014, item 1408) [17]. geographic coordinates of study sites of nowosady village (53°00'09.5" n / 23°13'00.6" e – 53°00'21.6" n / 23°13'45.4" e) were established by gps. results and discussion diversity of lichens out of 40 taxons identified in the study area, 39 were identified to the species level. the presented list of lichen species was ordered alphabetically. symbols of trees and bushes: trees: ah – aesculus hippocastanum, ap – acer platanoides, bp – betula pendula, ca – corylus avellana, pa – picea abies, ps – pinus sylvestris, pt – populus tremula, qr – quercus robur, ra – robinia pseudoaccacia; sa – salix alba, tc – tilia cordata; fruit and ornamental trees: c – cerasus sp., m – malus sp., p – prunus sp., pav – prunus avium, py – pyrus sp.; bushes: rt – rhus typhina, sv – syringa vulgaris. strictly protected species are indicated by “!!”, partly protected – by “!” and threatened by: en – endangered, nt – near threatened, vu – vulnerable. bryoria fuscescens (gyeln.) brodo & d. hawksw. – on wooden fence, !, vu; buellia punctata (hoffm.) a. massal. – on bark of tree (bp, m), wooden fence; caloplaca citrina (hoffm.) th. fr. s.l. – on concrete foundation; caloplaca decipiens (arnold) blomb. & forssell – on concrete electric poles and fence; candelaria concolor (dicks.) stein – on bark of tree (c); 3 of 5© the author(s) 2015 published by polish botanical society acta mycol 50(1):1053 kiercul / species richness of lichens in the nowosady village candelariella aurella (hoffm.) zahlbr. – on concrete electric poles and fence; candelariella xanthostigma (ach.) lettau – on bark of tree (bp, p, qr, ra, tc); cladonia chlorophaea (sommerf.) spreng. s.l. – on wooden fence; cladonia fimbriata (l.) fr. – on bark of tree (ps, tc), wooden fence; evernia prunastri (l.) ach. – on bark of tree (c, qr, tc), wooden fence, nt; hypocenomyce scalaris (ach.) m. choisy – on bark of tree (ps), wooden fence; hypogymnia physodes (l.) nyl. – on bark of tree (ah, ap, bp, c, p, pa, ps, pav py, qr, sv, rt), wooden fence; hypogymnia tubulosa (schaer.) hav. – on bark of tree (qr), !, nt; lecanora albescens (hoffm.) flörke s.l. – on concrete electric poles and fence; lecanora carpinea (l.) vain. – on bark of tree (p, qr); lecanora chlarotera nyl. – on bark of tree (p) – on roadside tree; lecanora conizaeoides cromb. – on bark of tree (ps), wooden fence; lecanora dispersa (pers.) sommerf. s.l. – on concrete electric poles and fence; lecanora muralis (schreb.) rabenh. – on erratic boulder, concrete electric poles and on eternit roof slates; lecidella stigmatea (ach.) hertel & leuckert – on concrete electric poles; lepraria sp. – on bark of tree (ah, p, ra); melanohalea exasperatula (nyl.) o. blanco et al. – on bark of tree (ah, c, p, qr), wooden fence; melanelixia fuliginosa (duby) o. blanco et al. – on wooden fence; parmelia sulcata taylor – on bark of tree (ah, ap, bp, m, p, qr, rt, tc), wooden fence; phaeophyscia orbicularis (neck.) moberg – on bark of tree (ah, m, pt, sa, rt, tc), wooden fence, concrete fence, steel, eternit roof slates; phaeophyscia nigricans (flörke) moberg – on concrete fence; phlyctis argena (spreng.) flot. – on bark of tree (ap, ca, qr, ra, sa); physcia adscendens h. olivier – on bark of tree (ah, ap, m, p, pt, py, rt, sa), steel; physcia caesia (hoffm.) fürnr. – on erratic boulder, concrete electric poles and fence, on steel; physcia dubia (hoffm.) lettau var. dubia – on bark of tree (ah, m, p, tc); physcia stellaris (l.) nyl. – on bark of tree (p, pav), wooden fence; physcia tenella (scop.) dc. – on bark of tree (m, p, rt) and on steel; physconia enteroxantha (nyl.) poelt – on bark of tree (ah), wooden fence; pseudevernia furfuracea (l.) zopf – on bark of tree (c); ramalina farinacea (l.) ach. – on bark of tree (tc), !, vu; ramalina fraxinea (l.) ach. – on bark of tree (qr), wooden fence, !!, en; scoliciosporum chlorococcum (stenh.) vězda – on bark of tree (tc); trapeliopsis granulosa (hoffm.) lumbsch – on wooden fence; xanthoria parietina (l.) th. fr. – on bark of tree (ap, bp, c, p, pa, pt, pav, py, m, pt, qr, rt, sa, sv), wooden fence, on concrete electric poles and fence, on steel; xanthoria polycarpa (hoffm.) rieber – on bark of tree (bp), wooden fence. the dominant group of lichens comprises species with crustose thalli (38%), which inhabit shaded areas. some of them colonize smooth tree bark, substrates of anthropogenic origin or soil. similarly represented are foliose lichens (38%), which prefer more sunny locations. lichens with dimorphous thalli (5%) live on wooden fences. numerous are also the species with fruticose thalli (13%), inhabiting tree bark. the percentage of lichens representing other morphological groups (placodial and squamulose) is minor, reaching nearly 6%. habitat preferences of lichens epiphytes. out of 40 lichen species found in the study area, 27 are epiphytes colonizing the bark of trees (27 tree species) or shrubs (6 species). eleven species are the so-called exclusive epiphytes growing on a variety of tree species including: q. robur (9 species), a. hippocastanum (8), t. cordata (8), a. platanoides (6), b. pendula (6), p. sylvestris (4), s. alba (4), p. tremula (3), r. pseudoacacia (3), p. abies (2), c. avellana (1), fruit and ornamental trees: prunus sp. (12), malus sp. (7), cerasus sp. (6), p. avium 4 of 5© the author(s) 2015 published by polish botanical society acta mycol 50(1):1053 kiercul / species richness of lichens in the nowosady village (3), pyrus sp. (3), and bushes – rh. typhina (6), s. vulgaris (2). clearly visible is the large proportion of nitrophilous lichens with large thalli [18], representing the genera: physcia (p. adscendens, p. dubia, p. stellaris and p. tenella), physconia (p. enteroxantha), ramalina (r. farinacea, r. fraxinea) or xanthoria (x. parietina, x. polycarpa). some rare lichen species were detected on the bark of these trees: ramalina farinacea and r. fraxinea. moreover, several species like: candelariella xanthostigma, phaeophyscia enteroxantha, physcia adscendens, p. tenella, xanthoria parietina and x. polycarpa represent mesophytic or xerophytic character [19]. analysis of epiphytic species in nowosady village has shown, that the number of 27 identified lichen species, is similar to that detected in other small villages in poland, e.g., 31 taxons found in klewinowo [10], 26 in hermanówka [11] and 33 epiphytes from świdnik town [3]. similar richness of epiphytes was observed in orchards in slovakia (52 epiphytic lichen species) and italy (45). in polish orchards 32 epiphytes were noted [8]. lower than in other studies epiphytic species richness found in poland can be partially explained by diverse geographical location of the study sites. climatic conditions, especially rainfall, are considered as the most important factors that impact the species diversity. in comparison with other parts of poland as well as with sites in italy and slovakia, nowosady village is characterized by smallest amount of rainfall [20]. this kind of climatic conditions would negatively influence the growth of lichens. the number of species established in nowosady is poorer than in białowieża (94 epiphytes lichen species) [7], olsztyn (63) [2], boćki (57) [6], mielnik (47) [5] and drohiczyn (43) [4]. epixylic lichens. the next most abundant ecological group found in the study area (18 species) consisted of lichens thriving on dead wood. four of them are exclusively epixylic. in nowosady village and in the surrounding areas, the epixylic lichens are closely associated with anthropogenic wood substrates (pillars, fences, crosses, utility buildings). matwiejuk [6], for example, has observed 41 taxons from epixylic lichens in boćki. in other towns and small localities on podlaskie province different number of epixylic lichens was observed: 26 species in mielnik [5], 24 species in białowieża [7], and only six species in drohiczyn [4]. kiercul [10] has observed 26 species representing this type of lichens in klewinowo and 12 in hermanówka [11]. epilithes and lichens of atypical substrates. the epilithic lichen biota was represented by a total of 12 species. epilithic species grow on natural substrates (glacial erratics, rocks) and anthropogenic surfaces (concrete pillars, mortared walls, tombstones). exclusively epilithic lichens identified in the study area were physcia caesia and lecanora muralis. eleven species from this group was also found to colonize artificial substrates (concrete, bricks). they included caloplaca citrina, c. decipiens, lecanora albescens, or xanthoria parietina. in the present study these calciphilous lichens accompany nitrophilous species, chiefly from the family physciaceae. total species richness of epilithic lichens found in nowosady (12 species) appeared comparable to the number of species detected previously in other small villages in poland: 17 taxons in klewinowo [10] 9 taxons in hermanówka [11] and 17 taxons in polish town – świdnik. in contrast higher species richness of epilithic lichens was noted in boćki [6] and mielnik [5] (38 species) as well as in drohiczyn [4] and białowieża [7] (32 and 24 species respectively). an interesting phenomenon of the present study is the tendency for colonization of atypical substrates such as eternit roof slates by lichen species lecanora muralis and phaeophyscia orbicularis. previously kiercul [10] has observed 15 taxons colonizing atypical substrates in klewinowo, and only one lichen lecanora muralis in hermanówka [11]. nowosady village has a fairly large number of old buildings and wooden fences which offer comfortable living conditions to lichens preferring and colonizing organic substrates. the limited number of natural habitats in the form of rocks or glacial erratics also causes intensive growth of calciphilous lichens colonizing anthropogenic substrates. obtained results are expected to contribute to the advance of more lichen oriented protection strategies. 5 of 5© the author(s) 2015 published by polish botanical society acta mycol 50(1):1053 kiercul / species richness of lichens in the nowosady village acknowledgments i would like to thank prof. maria rudawska for valuable discussions and guidance. references 1. fałtynowicz w, izydorek i, budzbon e. the lichen flora as bioindicator of air pollution of gdańsk, sopot and gdynia. łódź: polish botanical society; 1991. (monographiae botanicale; vol 73). http://dx.doi.org/10.5586/mb.1991.001 2. kubiak d. lichens of red oak quercus rubra in the forest environment in the olsztyn lake district (ne poland). acta mycol. 2006;41(2):319–328. http://dx.doi.org/10.5586/ am.2006.033 3. wójciak h, korona k. the condition of the biota of lichens in świdnik. teka komisji ochrony i kształtowania środowiska przyrodniczego. 2008;5:199–207. 4. matwiejuk a. lichens of drohiczyn on the bug river (podlasie, eastern poland). rocz ar pozn bot stec. 2009;13:57–62. 5. matwiejuk a. lichens of mielnik on bug river (podlasie, eastern poland). opole scientific society nature journal. 2008;41:5–18. 6. matwiejuk a. lichens of the boćki and its surroundings in podlasie (ne poland). opole scientific society nature journal. 2009;42:49–61. 7. matwiejuk a. anthropogenic changes of lichen biota of the białowieża town (podlasie, eastern poland). rocz ar pozn bot stec. 2011;15:129–138. 8. zarabska d, guttova a, cristofolini f, giordani p, lackovicova a. epiphytic lichens of apple orchards in poland, slovakia, and italy. acta mycol. 2009;44(2):151–163. http:// dx.doi.org/10.5586/am.2009.013 9. szymczyk r, zalewska a. lichens in the rural landscape of the warmia plain. acta mycol. 2008;43(2):215–230. http://dx.doi.org/10.5586/am.2008.026 10. kiercul s. species diversity of lichens (fungi lichenisati) in klewinowo village and surrounding areas (ne poland). parki narodowe i rezerwaty przyrody. 2013;32(4):77–94. 11. kiercul s. species richness of lichens in the hermanówka village. chrońmy przyr ojcz. 2015;71(2):116–121. 12. kondracki j. geografia regionalna polski. warszawa: pwn; 2013. 13. cieśliński s, fałtynowicz w, editors. atlas of geographical distribution of lichens in poland. cracow: w. szafer institute of botany, polish academy of sciences; 1993. (vol 1). 14. diederich p, ries c. lichens of belgium, luxembourg and northern france [internet]. 2014 [cited 2014 sept 30]; available from: http://www.lichenology.info 15. blanco o, crespo a, divakar pk, esslinger tl, hawksworth dl, lumbsch ht. melanelixia and melanohalea, two new genera segregated from melanelia (parmeliaceae) based on molecular and morphological data. mycol res. 2004;108(8):873–884. http://dx.doi. org/10.1017/s0953756204000723 16. cieśliński s, czyżewska k, fabiszewski j. red list of the lichens in poland. in: mirek z, zarzycki k, wojewoda w, szeląg z, editors. red list of plants and fungi in poland. cracow: w. szafer institute of botany, polish academy of sciences; 2006. p. 71–90. 17. regulation of the minister of the environment dated october 9, 2014, on the protection of species of fungi (oj from 2014, item 1408). 18. wirth v. zeigerwerte von flechte. 3. aufl. scripta geobotanica. 2001;18:221–243. 19. nimis pl, martellos s. italic – the information system on italian lichens. version 4.0 [internet]. 2008 [cited 2015 feb 17]; available from: http://dbiodbs.univ.trieste.it 20. farata r, editor. atlas klimatu województwa wielkopolskiego. poznań: imgw; 2004. http://dx.doi.org/10.5586/mb.1991.001 http://dx.doi.org/10.5586/am.2006.033 http://dx.doi.org/10.5586/am.2006.033 http://dx.doi.org/10.5586/am.2009.013 http://dx.doi.org/10.5586/am.2009.013 http://dx.doi.org/10.5586/am.2008.026 http://www.lichenology.info http://dx.doi.org/10.1017/s0953756204000723 http://dx.doi.org/10.1017/s0953756204000723 http://dbiodbs.univ.trieste.it abstract introduction study area material and methods results and discussion diversity of lichens habitat preferences of lichens acknowledgments references 2015-08-02t16:08:02+0100 piotr otręba the third report of colletotrichum japonicum worldwide acta mycologica article id: 565 doi: 10.5586/am.565 publication history received: 2020-04-30 accepted: 2021-02-01 published: 2021-05-20 handling editor małgorzata ruszkiewicz-michalska; institute for agricultural and forest environment, polish academy of sciences; university of łódź, poland; https://orcid.org/00000001-8901-0552 funding the study was supported by the department of botany, mycology and ecology, maria curie-skłodowska university in lublin through its statutory funds. competing interests no competing interests have been declared. copyright notice © the author(s) 2021. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. short communication in plant pathology the third report of colletotrichum japonicum worldwide urszula świderska-burek * department of botany, mycology and ecology, maria curie-skłodowska university, poland * to whom correspondence should be addressed. email: urszula.swiderska-burek@poczta.umcs.lublin.pl abstract is paper reports the first finding of colletotrichum japonicum (hemmi) bedlan on berberis aquifolium pursh [= mahonia aquifolium (pursh) nutt.] in poland. e fungus was collected in the botanical garden of maria curie-skłodowska university (umcs) in lublin. is is the second locality of the species in europe and the third worldwide. in this paper, morphological characteristics, microphotographs, and distribution comments are provided. keywords anthracnose; coelomycetous anamorphs; distribution; microfungi; oregon grape 1. introduction colletotrichum is traditionally recognized as an asexual genus of fungi, with a number of species linked to sexual morphs assigned to the glomerella genus (sordariomycetes, glomerellaceae, ascomycota). over the years, colletotrichum species were considered to be host specific, which led to the description of a large number of taxa (cannon et al., 2012; sharma & shenoy, 2016). species from this genus are economically one of the most important fungi, and cause anthracnose and other diseases in a wide range of plant species (cai et al., 2009; hyde et al., 2009; sutton, 1980). e comprehensive monographic study conducted by von arx (1957) based on morphological characteristics with little emphasis placed on pathological features led to drastic reduction from approximately 750 to 11 species. molecular studies of colletotrichum species, which have become an integrated element in taxonomic research, have been developing intensively since the end of the twentieth century. e first applications of dna sequence data to distinguish between species within this genus used the internal transcribed spacer (its1) region of nrdna. later, multilocus analyses became a common approach based on the use of its2 and large ribosomal subunit (lsu) as well as partial sequences of genes encoding, e.g., histone h3 (his), translation elongation factor 1 alpha (tef-1α), tubulin beta chain (tub), and actin (act) (cannon et al., 2012). scientists working on colletotrichum do not agree on the taxonomic approach and characters that should be employed to identify and describe a new colletotrichum species. hence, a reliable secondary barcode marker is indispensable for the accurate identification of colletotrichum species (sharma & shenoy, 2016). identification of colletotrichum species based on morphology has always been problematic; therefore, the current progress in molecular phylogenetic methods facilitates identification of stable and well-separated clades within colletotrichum. however, the taxonomy of this genus is still unsatisfactory and there is a need for a polyphasic approach for identification, which reflects the natural classification of species and subspecific taxa within the genus (cai et al., 2009). berberis aquifolium (berberidaceae) is a plant native to western north america. is species was introduced to europe as an ornamental plant in the 1820s. acta mycologica / 2021 / volume 56 / article 565 publisher: polish botanical society 1 https://doi.org/10.5586/am.565 https://orcid.org/0000-0001-8901-0552 https://orcid.org/0000-0001-8901-0552 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0002-3120-276x mailto:urszula.swiderska-burek@poczta.umcs.lublin.pl świderska-burek / colletotrichum japonicum on berberis aquifolium – third locality worldwide table 1 features of colletotrichum species reported to be associated with berberis aquifolium leaves. c. fioriniae (marcelino & gouli) pennycook c. japonicum (hemmi) bedlan c. mahoniae fabric. c. nymphaeae (pass.) aa leaf spots up to 10 mm 1–5 mm 1–1.5 mm 0.5–5 mm irregularly circular mostly irregularly shaped rounded roundish brown, with a margin, surrounded by a chloric halo first greyish-brown, finally grey grey pale brownish, greyish-brown with a dark reddish-brown to purplish-brown margin, later dark brown to black conidia 9.9–14 × 3–4.3 μm∗ 10.65–16.83 × 5.08–7.51 μm 12–12.5 × 4.5–5 μm 11.8–23 × 4.2–5.6 μm mostly ellipsoidal, short cylindrical or ovoid cylindrical ellipsoidal or cylindrical ends rounded hyaline, two-punctated or granulated rounded at the apex, attenuated, occasionally truncate at the base setae no data very few if present then occur abundantly, with swelled basis not formed references garibaldi et al., 2020 bedlan, 2012 fabricatore, 1950 van der aa, 1978 ∗ measurements based on in vitro cultures. at present, it grows in many parts of europe (sorokopudov et al., 2017). it was introduced in poland in 1839, and the status of this species has changed from a crop plant to a locally established and invasive species (tokarska-guzik et al., 2012). according to a preliminary checklist of micromycetes in poland, 23 colletotrichum species were reported in poland up until 2008 (mułenko et al., 2008), while 26 species are currently known (farr & rossman, 2020; jayawardena et al., 2016; okorski et al., 2018; pszczółkowska et al., 2016, 2017). e present paper provides information about a new colletotrichum species for polish funga (c. japonicum on b. aquifolium); it also reports the second locality of the fungus in europe and third worldwide. 2. material and methods e infected leaves of b. aquifolium were collected in the botanical garden of maria curie-skłodowska university (umcs) in lublin in 2018. e material sample was stained with cotton blue in lactic acid and gently heated. observations were made using an olympus szx10 stereoscopic microscope and an olympus cx31 light microscope. photographic documentation was taken with an olympus xc50 microscope camera. measurements were made at ×600 and/or ×300 magnifications. structure size ranges were specified based on 10 measurements of acervuli, 15 setae, and 30 conidia. e publications mentioned in table 1 were used for identification of the fungus. e name of the host plant and its synonym is accepted following e plant list (http://www.theplantlist.org/). e examined specimen is deposited in the herbarium of maria curie-skłodowska university in lublin (lbl). 3. results colletotrichum japonicum (hemmi) bedlan, j. kulturpfl. 64(12): 478–481. 2012 (figure 1) leaf spots mostly elliptical, roundish, or irregular, 2–6 mm in diameter, greyish-brown to greyish with blackish-brown margin. acervuli within the spots, epiphyllous, 112–300 × 90–180 μm, oen with setae. setae short, black-brown, straight or slightly curved, 40–55 × 3–3.5 μm. conidia one-celled, hyaline, elliptical or ovoid, straight or slightly curved, rounded at the ends, 12.5–17 × 4.5–6.5 μm. acta mycologica / 2021 / volume 56 / article 565 publisher: polish botanical society 2 http://www.theplantlist.org/ świderska-burek / colletotrichum japonicum on berberis aquifolium – third locality worldwide figure 1 colletotrichum japonicum on berberis aquifolium (lbl m–32789). (a) leaf spots with visible acervuli. (b) acervuli. (c) acervulus with setae. (d) conidia. scale bars: (a) 15 mm; (b) 200 μm; (c,d) 50 μm. structures in a microscopic preparation stained with cotton blue in lactic acid. photographs: u. świderska-burek. specimen examined: on berberis aquifolium pursh (berberidaceae). poland: lublin – botanical garden of the maria curie-skłodowska university, september 25, 2018, leg. u. świderska-burek (lbl m–32789). 4. comments e analyzed polish sample exhibits the features of c. japonicum provided in the literature from austria (table 1), i.e., the only locality worldwide where the fungus has been noted on the same host (b. aquifolium) (bedlan, 2012). on b. aquifolium, three other colletotrichum species have been reported to date. all of them, c. nymphaeae (damm et al., 2012), c. mahoniae (fabricatore, 1950), and c. fioriniae (garibaldi et al., 2020), were reported from italy. additionally, two species were recorded on another host species from the mahonia genus (currently assigned to the berberis genus), i.e., m. bealei (fortune) pynaert: c. mahoniae from georgia and c. gloeosporioides penz. (probable host m. bealei incorrectly reported as m. healei) from china (bedlan, 2012; farr & rossman, 2020). acknowledgments i would like to thank dr. grażyna szymczak, the director of the botanical garden of umcs in lublin, for facilitating the research, notably the collection of plants infected by the fungus. acta mycologica / 2021 / volume 56 / article 565 publisher: polish botanical society 3 świderska-burek / colletotrichum japonicum on berberis aquifolium – third locality worldwide references bedlan, g. (2012). mahonia aquifolium – eine neue wirtspflanze von colletotrichum japonicum comb. nov. [mahonia aquifolium – a new host plant of colletotrichum japonicum comb. nov.]. journal für kulturpflanzen, 64(12), 478–481. https://doi.org/10.5073/jfk.2012.12.04 cai, l., hyde, k. d., taylor, p. w. j., weir, b., waller, j., abang, m. m., zhang, j. z., yang, y. l., phoulivong, s., liu, z. y., prihastuti, h., shivas, r. g., mckenzie, e. h. c., & johnston, p. r. 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(2012). rośliny obcego pochodzenia w polsce ze szczególnym uwzględnieniem gatunków inwazyjnych [alien plants in poland with particular reference to invasive species]. generalna dyrekcja ochrony środowiska. van der aa, h. a. (1978). a leaf spot disease of nymphaea alba in the netherlands. netherlands journal of plant pathology, 84, 109–115. https://doi.org/10.1007/bf01981538 von arx, j. a. (1957). die arten der gattung colletotrichum cda [e species of the genus colletotrichum cda]. phytopathologische zeitschri, 29(4), 413–468. acta mycologica / 2021 / volume 56 / article 565 publisher: polish botanical society 4 https://doi.org/10.5073/jfk.2012.12.04 https://doi.org/10.3114/sim0014 https://doi.org/10.3114/sim0010 https://nt.ars-grin.gov/fungaldatabases/ https://doi.org/10.1094/pdis-10-19-2104-pdn https://doi.org/10.5943/mycosphere/si/2c/1 https://doi.org/10.1094/pdis-12-17-2023-pdn https://doi.org/10.1094/pdis-04-16-0425-pdn https://doi.org/10.1094/pdis-04-17-0471-pdn https://doi.org/10.5943/mycosphere/si/2c/2 https://doi.org/10.1007/bf01981538 the third report of colletotrichum japonicum worldwide 1 introduction table 1 2 material and methods 3 results figure 1 4 comments acknowledgments references suillus lakei (murrill) a. h. sm. & thiers (boletales, basidiomycota) in poland: new data 1 of 6published by polish botanical society acta mycologica short communication suillus lakei (murrill) a. h. sm. & thiers (boletales, basidiomycota) in poland: new data andrzej szczepkowski1*, tomasz olenderek2 1 division of mycology and forest phytopathology, department of forest protection and ecology, faculty of forestry, warsaw university of life sciences – sggw, nowoursynowska 159, 02-776 warsaw, poland 2 department of geomatics and land management, faculty of forestry, warsaw university of life sciences – sggw, nowoursynowska 159, 02-776 warsaw, poland * corresponding author. email: andrzej_szczepkowski@sggw.pl abstract this study presents up-to-date data on the distribution of the western painted suillus suillus lakei in poland. this alien species of fungus was first recorded in poland in 2012, followed by a second recording in 2013. the third site, presented in this article, was discovered in 2016. a description of the newly discovered site and the basidiomata is given. all three sites of s. lakei are located on private properties overgrown with young douglas fir. two of the three recorded sites are located within the boundaries of the wielkopolskie province, and one in the kujawsko-pomorskie province. further expansion of this fungus in poland is expected. keywords alien species; western painted suillus; macrofungi; ectomycorrizal fungi; douglas fir; pseudotsuga menziesii; distribution in poland introduction suillus lakei (murrill) a. h. sm. & thiers is an example of an additional alien species of mycobiota – besides agaricus subrufescens peck [1,2] and aureoboletus projectellus (murrill) halling [3] – recorded in poland in recent years. this fungus is known under a number of synonymous names: boletus lakei murrill, boletinus lakei (murrill) singer, ixocomus lakei (murrill) singer, and suillus amabilis (peck) singer [4]. the epithet lakei was added in recognition of a mycologist from the university of oregon, e. r. lake [5]. in northwestern north america – where this suillus originates – it is one of the common species of bolete fungi. suillus lakei is well known as ectomycorrhizal with the douglas fir, and its basidiomata can be found on the ground in summer and fall amidst the grass of young coniferous forests as well as in parks and gardens with douglas fir. basidiomata of s. lakei appear in large numbers especially after the first abundant fall rainfalls [5,6]. in north america, basidiomata of s. lakei are commonly observed in direct vicinity of spike-cap gomphidius subroseus kauffman, which is probably a parasite on the mycorrhizal mycelium of the western painted suillus [7]. together with its partner-tree (douglas fir, pseudotsuga menziesii), s. lakei was introduced to south america [8], new zealand [9], and europe, where it was recorded in bosnia and herzegovina, bulgaria, the czech republic, denmark, germany, slovakia, hungary, great britain, and italy [10–12]. since 2012, s. lakei is known also from poland [13,14], where it has also been identified from ectomycorrhizas by pietras et al. [15]. suillus lakei is relatively easy to recognize by its appearance in the vicinity of the douglas fir and on the basis of its pileus with a diameter ranging from 3 to 15(18) cm (usually dry, but in older specimens sticky during moist weather), covered with clearly doi: 10.5586/am.1098 publication history received: 2017-05-08 accepted: 2017-10-22 published: 2017-11-20 handling editor tomasz leski, institute of dendrology, polish academy of sciences, poland authors’ contributions as: identification of the specimens, writing of the manuscript, preparation of the map; to: field research, correction of the manuscript funding publishing of the manuscript was covered by the statutory funds of the faculty of forestry, warsaw university of life sciences – sggw. competing interests as is a member of the editorial council of the acta mycologica; to: no competing interests copyright notice © the author(s) 2017. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation szczepkowski a, olenderek t. suillus lakei (murrill) a. h. sm. & thiers (boletales, basidiomycota) in poland: new data. acta mycol. 2017;52(2):1098. https://doi. org/10.5586/am.1098 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:andrzej_szczepkowski%40sggw.pl?subject=suillus%20lakei%20%28murrill%29%20a.%20h.%20sm.%20%26%20thiers%20%28boletales%2c%20basidiomycota%29%20in%20poland%3a%20new%20data https://doi.org/10.5586/am.1098 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1098 https://doi.org/10.5586/am.1098 2 of 6© the author(s) 2017 published by polish botanical society acta mycol 52(2):1098 szczepkowski and olenderek / suillus lakei in poland: new data visible and usually rust-colored, reddish-brown bundles of fibers or scales (sometimes absent in older specimens) (e.g., [16–19]. within this species, four varieties are distinguished. according to klofac [20], the dimensions of the spores of the nominate variety s. lakei var. lakei reach 11 × 3.7(–4) µm. the variety s. lakei var. pseudopictus, recorded in north america (and in europe known only from italy), differs from the nominate variety only in its smaller (up to 10 cm wide), darker, more brick-red, rougher pileus and its – according to some authors – somewhat smaller (9 × 4 µm) spores [16,20–22]. in italy, the variety s. lakei var. calabrus, which is characterized by its brighter, yellowish cap covered by rusty red scales, was recorded under douglas firs growing in acidic soils [10]. on the other hand, s. lakei var. landkammeri is characterized by the olive hue of its pores, its hymenophore tubes that visibly descend onto the stipe, and its spores of up to 4.8 µm in width. the western painted suillus is considered an edible species, especially young mushrooms, although opinions differ significantly regarding its taste qualities [22]. a significant amount of alkaloids and tannins is found in the basidiomata of s. lakei, suggesting also the potential medicinal properties of this fungus [23]. the aim of this work is to summarize the data regarding previous observations and distributions of localities of s. lakei in poland and to present a newly discovered site and a description of the basidiomata discovered there. material and methods the list of documented sites of s. lakei basidiomata occurrence in poland is presented according to physico-geographical divisions – by subprovince, macroregion, and microregion [24] – and polish administrative divisions at the level of province and county. the names of fungi are cited according to mycobank [4], and plants after mirek et al. [25]. observations and measurements of 30 spores were conducted in water using a nikon eclipse e400 microscope. results previously, only two localities of s. lakei in poland have been reported. the locality presented in this work is the third site in poland. all localities are found on private properties situated within the boundaries of three mesoregions: lower vistula valley (polish: dolina dolnej wisły), great poland lakeland (polish: pojezierze wielkopolskie), and southern great poland lowland (polish: nizina południowowielkopolska) and two provinces: kujawy-pomerania and great poland. list of localities of suillus lakei in poland (fig. 1) new locality. central polish lowlands (polish: niziny środkowopolskie), southern great poland lowland, kalisz upland (polish: wysoczyzna kaliska): brudzew (kalisz county, great poland province), on private property, in grass in the vicinity of young douglas firs, on november 5, 2016, leg. irena & józef bażant, det. andrzej szczepkowski. localities known from the literature. south baltic lakeland (polish: pojezierze południowobałtyckie), lower vistula valley, fordon valley (polish: dolina fordońska): pruszcz pomorski (świecie county, kujawy-pomerania province), october 8, 2012 [13]; south baltic lakeland, fig. 1 distribution of suillus lakei (murrill) a. h. sm. & thiers in poland (white circle: new locality; black circles: localities known from the literature). 3 of 6© the author(s) 2017 published by polish botanical society acta mycol 52(2):1098 szczepkowski and olenderek / suillus lakei in poland: new data great poland lakeland, września plain (polish: równina wrzesińska): poznań-szczepankowo (poznań county, great poland province), september 28 and october 10, 2013 [14] (and personal communication, 2017). description of the new locality and basidiomata in brudzew on november 5, 2016, a site of s. lakei was found within the locality of brudzew (blizanów municipality, kalisz county, great poland province). the site is located on private property (n 51°55'35"; e 17°59'10") encompassing approximately 0.7 hectares. apart from a residential building and utility buildings, the greater part of the property’s terrain is occupied by an orchard with fruit trees and shrubs (apples, plums, raspberries, blackberries, and grapes), english oaks quercus robur and several species of coniferous trees: douglas fir pseudotsuga menziesii subs. menziesii, european larch larix decidua var. polonica, scots pine pinus sylvestris and ponderosa pine p. ponderosa, norway spruce picea abies and blue spruce p. pungens, and northern white cedar thuja occidentalis. currently, more than 30 trees of douglas fir are growing on the property. the oldest tree is about 20 years old and the youngest about 4–5 years old. according to the owners of the property, the seedlings of the douglas firs originate from the beskid mountains (wisła forest district). basidiomata of s. lakei were observed in the direct vicinity of the apple trees, northern white cedar, pines, spruces, and two young douglas firs (70 cm height). among nine basidiomata of s. lakei; seven were characterized by fused stipes and two were growing individually in grass, at a distance of 3–4 meters from the douglas firs. the diameter of the pilei ranged from 3 to 12 cm and a thickness reached 2.5 cm (the tube layer was up to 0.9 cm). the surface of the pileus was covered with dry, rough, rusty reddish-brown fibrils. the tubes and pores were olive-yellow. the pores were angular and elongated, up to 2.5 mm wide, partially descending onto the stipe. when pressed, the pores got darken. the stipes were full, cylindrical, ochre in color, and from 3 to 9 cm in height and 0.8 to about 3 cm in width. on the stipes and at the edge of the younger pilei, the remnants of the whitish partial veil were visible. the tissue of the pileus and stipe was yellowish with a subtle orange hue; without changing a color when exposed to the air. there was no perceptible smell; the taste was somewhat acidic. the spores were smooth, elongated ellipsoids with dimensions (9–)10–11(–12) × 3.5–4.5 µm, with one or a few drops. the spore print was olive-brown. apart from the s. lakei, basidiomata of greville’s bolete s. grevillei were also growing on the property, in the vicinity of larches. dried basidiomata of s. lakei were preserved in the fungarium of the division of mycology and forest phytopathology of the warsaw university of life sciences – sggw (waml). discussion douglas firs began to be introduced into europe (and into poland) in the first half of the nineteenth century [26]. the first record of s. lakei was reported after nearly 100 years from the first introduction of the douglas firs to europe and originate from denmark from 1920. the second and the last (so far) record of this fungus has been reported from that country in 1980 [27,28]. in 1949, s. lakei was found in czechoslovakia and was described as boletinus tridentinus (bres.) subsp. landkammeri by pilát and svrček [29]. few years later, chinková and pouzar [30] demonstrated that the described taxon is in fact the north american species boletinus lakei (murr.) sing. it has also been found in the czech republic in recent years [31]. in england, s. lakei was found once, in 1952 [32]. in other countries such as germany, s. lakei was noted for the first time in 1971 and also in recent years [33–36]. in 1989, it was found in hungary [37]. in italy, s. lakei was encountered since the end of the twentieth century [10,38,39]. in 2004, s. lakei was reported in bulgaria (currently known from a few stands) [11,40,41]. the first documented record of s. lakei in poland originates from 2012 and, because this fungus was earlier known in the neighboring countries (czech republic, slovakia, northeastern germany), we presume that s. lakei could also arrive to poland much earlier. in 1960, in her monograph on boletoid fungi of poland, skirgiełło included 4 of 6© the author(s) 2017 published by polish botanical society acta mycol 52(2):1098 szczepkowski and olenderek / suillus lakei in poland: new data a description of boletinus lakei (murr.) sing. (there with the polish name “borowiec lakego”) and information that the mushroom was not yet confirmed in poland but might possibly be found [42]. if we assume that s. lakei indeed arrived prior to 2012, then this brings up the question why this fungus had not been noticed before from the area of poland. the most probable explanation is that the basidiomata of s. lakei were rarely encountered by mushroom pickers. additionally, basidiomata were probably not distinguished from those of suillus cavipes = boletinus cavipes (whose basidiomata are characterized by a hollow stipe and mycorrhizal symbiosis with the larch larix) or even from some boletes xerocomus spp. however, we may also not exclude that the findings of s. lakei in poland in recent years roughly reflect the time of its arrival to poland. this assumption is supported by the fact that the fungus was only recently discovered in bulgaria and hungary. it is also possible that in poland s. lakei mycelium existed for a long time only in mycorrhizal form and from the unknown reason did not expressed in the form of basidiomata. mycorrhizas of s. lakei were recently reported by pietras et al. from young forest plantations of douglas fir [15]. the new questions concern factors influencing the appearance of basidiomata of s. lakei in recent years and potential further spreading of this fungus on the area of poland. therefore, more research is required on the occurrence of basidiomata of s. lakei on new plantings of douglas fir in gardens, parks, and forest plantations to better understand the biology of this alien fungus species and its expansion dynamics in poland. acknowledgments the authors express their sincere thanks to irena and józef bażant for conveying the basidiomata and information regarding the site of suillus lakei in brudzew and to prof. tadeusz tylkowski from the institute of dendrology, polish academy of sciences in kórnik for additional information regarding the locality in poznań-szczepankowo. the text was translated by bogna j. gladdenobidzińska and proofread by matthew e. gladden. references 1. szczepkowski a, gierczyk b, kujawa a. greenhouses of botanical gardens as a habitat of alien and native macrofungi: a case study in poland. cent eur j biol. 2014;9(8):777–795. https://doi.org/10.2478/s11535-014-0310-5 2. szczepkowski a, gierczyk b, olenderek t. pierwsze stwierdzenie na otwartej przestrzeni pieczarki brązowej agaricus subrufescens peck w polsce. przegląd przyrodniczy. 2015;26(3):80–83. 3. wrzosek m, gorczak m, pawłowska j, wilk m. boletus projectellus (murrill) murrill in poland – a possible threat for a natural pine wood mycocoenosis? 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(flora polska. rośliny zarodnikowe polski i ziem ościennych). http://www.poggenstoehl.homepage.t-online.de/douglasienr.pdf http://www.tintling.de/pilzkalender/2009/suillus_lakei.jpg abstract introduction material and methods results list of localities of suillus lakei in poland description of the new locality and basidiomata in brudzew discussion acknowledgments references 2017-11-20t17:23:13+0000 piotr otręba chemical compounds of extracts from sarcodon imbricatus at optimized growth conditions 1 of 11published by polish botanical society acta mycologica original research paper chemical compounds of extracts from sarcodon imbricatus at optimized growth conditions katarzyna sułkowska-ziaja1*, agnieszka szewczyk1, joanna gdula-argasińska2, halina ekiert1, jerzy jaśkiewicz3, bożena muszyńska1 1 department of pharmaceutical botany, faculty of pharmacy, jagiellonian university medical college, medyczna 9, 30-688 kraków, poland 2 department of radioligands, faculty of pharmacy, jagiellonian university medical college, medyczna 9, 30-688 kraków, poland 3 faculty of health and medical science, andrzej frycz modrzewski kraków university, gustawa herlinga-grudzińskiego 1, 30-705 kraków, poland * corresponding author. email: katarzyna.sulkowska-ziaja@uj.edu.pl abstract the effect of carbon and nitrogen sources and initial ph and temperature of the medium on the mycelial growth of sarcodon imbricatus (l.) p. karst. in axenic liquid culture was investigated. the optimal composition of the medium was found to be: 5% fructose, 1% hydrolysate of casein, 1% yeast extract, and 0.3% kh2po4 at ph = 6 and incubation temperature of 20°c. in this condition the maximum biomass growth was observed, yielding 10.2 g l−1 of dry weight after 3-week of growth. the medium regarded as optimal for growth of s. imbricatus mycelium was used for the production of the biomass and further chemical analysis. the quantitative and qualitative composition of phenolic acids, fatty acids, and sterols were determined using chromatographic methods. the total content of phenolic acids was 1.86 mg × 100 g−1 dw, with the largest amount of protocatechuic acid (1.27 mg × 100 g−1 dw). nineteen fatty acids were estimated, including five unsaturated fatty acids, e.g., oleic and α-linolenic acid. the analysis of sterols composition revealed the presence of ergosterol and ergosterol peroxide (197.7 and 200.47 mg × 100 g−1 dw, respectively). these compounds were isolated and confirmed by 1h-nmr. presented study constitutes the first report on the accumulation of substances (phenolic acids, fatty acids, and sterols) with multidirectional biological activity in the mycelial axenic culture of sarcodon imbricatus. keywords sarcodon imbricatus; in vitro cultures; secondary metabolites introduction an alternative method to obtain secondary metabolites with therapeutic properties from fungal fruiting bodies is to use the biosynthetic capability of in vitro mycelial cultures. the main advantage of in vitro cultures is their independence from the environmental conditions and the ability to produce continuously high-quality material. the intensity of metabolites’ synthesis and their further accumulation can be controlled by the nutrient composition of the medium and the environmental parameters during culture. for this reason, it is important to optimize the conditions for in vitro culture of fungi [1,2]. the genus sarcodon includes some species with therapeutic potentials, e.g., sarcodon aspratus was found effective in antitumor activity and sarcodon scabrosus in anti-inflammatory actions [3–5]. interesting biological properties of this genus doi: 10.5586/am.1086 publication history received: 2016-04-29 accepted: 2016-12-15 published: 2016-12-27 handling editor maria rudawska, institute of dendrology, polish academy of sciences, poland authors’ contributions ksz collected, optimized, and examined sterols; as examined phenolic acids; jga examined fatty acids; all authors contributed to the manuscript preparation funding publishing of the manuscript was financially supported by the statutory funds of the jagiellonian university medical college. competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation sułkowska-ziaja k, szewczyk a, gdula-argasińska j, ekiert h, jaśkiewicz j, muszyńska b. chemical compounds of extracts from sarcodon imbricatus at optimized growth conditions. acta mycol. 2016;51(2):1086. http://dx.doi. org/10.5586/am.1086 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:katarzyna.sulkowska-ziaja%40uj.edu.pl?subject=chemical%20compounds%20of%20extracts%20from%20sarcodon%20imbricatus%20at%20optimized%20growth%20conditions http://dx.doi.org/10.5586/am.1086 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1086 http://dx.doi.org/10.5586/am.1086 2 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1086 sułkowska-ziaja et al. / chemical analysis of mycelial culture of sarcodon imbricatus are associated with the occurrence of ergosterol peroxide (3β-hydroxy-5α,8αepidioxyergosta-6,22-dien), which exhibits a series of activities such as antileukemic and antitumor [6] and inflammatory activity [7]. recent studies have demonstrated that sarcodon imbricatus is also important as an agent that stimulates neuronal growth and promotes brain health [8]. our previous studies of s. imbricatus indicated antimicrobial activity against five strains of gram-positive and gram-negative bacteria, antiviral activity against hpv-1 virus, and cytotoxic activity toward b16 murine melanoma tumor cell lines, sarcoma xc, and human breast cancer cell line in vitro of selected polysaccharide fractions isolated from mycelial cultures of this fungal species [9,10]. this mushroom is ectomycorrhizal species associated with picea, common in the south-west of europe, but becoming rare in poland [11]. fruiting bodies are considered edible, though it can have a slightly bitter taste and therefore the culinary value of s. imbricatus is considered moderate [12]. when eaten raw it can be even poisonous. the aim of the present study was to initiate in vitro culture of s. imbricatus to determine the optimal conditions for mycelial growth and accumulation level of compounds with biological activity in extracts from the obtained biomass. presented results are the first report on the quantitative and qualitative analysis of a group of compounds with multidirectional therapeutic effects such as phenolic acids, fatty acids, and sterols obtained from the biomass of mycelial cultures of s. imbricatus. these compounds show various biological activities such as antitumor, immunostimulating, antioxidant, and protection of cardiovascular system [13]. material and methods mushroom material and obtaining in vitro culture the fruiting bodies of s. imbricatus were collected under spruce trees (picea sp.) in the forest district of krynica (southern poland) in august 2008. four fruiting bodies were found and after taxonomic identification [14,15], one of them was used to obtain mycelial cultures. representative voucher specimens were deposited at the department of pharmaceutical botany, pharmaceutical faculty, jagiellonian university medical college, kraków. the in vitro cultures were obtained from the hymenial part of the fruiting body. pieces of fruiting body were sterilized in 0.1% naocl for 10 min and inoculated on a petri dishes containing an agar-solidified standard medium (sm) of a composition described by turło [16] (glucose 5%, yeast extract 1%, casein hydrolysate 1%, kh2po4 0.3%, agar 10%). this initial cultures were grown at 22°c ±2°c and were sub-cultured every three weeks. the obtained strain ujcmbf-51mc (pcm 2719) was deposited in the polish collection of microorganisms (pcm), ludwik hirszfeld institute of immunology and experimental therapy, polish academy of sciences, wrocław, poland. after growing on the solid medium the pieces of mycelium were inoculated in the erlenmeyer flask (500 ml) containing 250 ml of the standard medium as described above and were maintained as agitated liquid cultures. optimization of culture conditions two types of liquid cultures, agitated and stationary ones were used for optimization of the culture conditions. an effect of carbon and nitrogen source and ph of the medium was evaluated in the agitated culture and an effect of temperatures was tested in stationary cultures to find the most beneficial carbon source for the growth of s. imbricatus mycelium, various carbon sources such as monosaccharides (fructose and glucose), disaccharides (maltose and sucrose), and polysaccharide (starch) were compared as a carbon source in a standard medium. 3 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1086 sułkowska-ziaja et al. / chemical analysis of mycelial culture of sarcodon imbricatus to analyze the effect of nitrogen sources on the mycelial growth, s. imbricatus mycelium was cultivated on a standard medium with casein hydrolysate, urea, ammonium sulfate, and ammonium phosphate as a nitrogen source. to test the effect of ph on growth of s. imbricatus mycelium, the ph value of the standard medium was adjusted from 3.0 to 7.5 at 0.5 increments using 0.1 m hcl or 0.1 m naoh. the ph values were checked using a digital ph-meter (cp-505 – elmetron, poland). the liquid cultures were maintained in an erlenmeyer flask (500 ml) containing 250 ml of a liquid medium. the culture was shaken at a rate of 140 rpm (shaker altel, poland), under 16-h light (900 lx / 8 dark) at 22°c ±2°c. the effect of temperature on the liquid stationary cultures was determined in an incubator (st500/b/40, pol-eko-aparatura, poland) at various temperatures (15°c, 20°c, 25°c, 30°c, and 35°c). cultures were maintained in an erlenmeyer flask (500 ml) containing 250 ml of a liquid standard medium. after 3-weeks of growth, the mycelium was separated from the liquid medium using a filter paper on a büchner funnel, rinsed with redistilled water, frozen and dried by lyophilization (lyophilizer freezone 4.5, labconco, usa; temperature: −40°c) and the dry biomass was determined. chemical analyzes the final composition of the culture medium selected during the optimization process was used for the cultivation of biomass for further chemical analysis. determination of phenolic acids. the extraction process was carried out as follows: 2  g of powdered lyophilized biomass was twice extracted with 100 ml of boiling methanol (at 67.4°c) for 2 h under a reflux condenser. the combined extracts (200 ml) were concentrated to dryness using a rotary vacuum evaporator at 40°c. then the residues were dissolved in 10 ml of methanol. the hplc method was followed according to the procedure developed by ellnainwojtaszek and zgórka with some modifications [17]. the qualitative and quantitative hplc analyzes were conducted using hplc vwr hitachi apparatus: autosampler l-2200, pump l-2130, rp18 column (250 × 4 mm, 5 µm) thermostated at 25°c, column oven l-2350, and diode array detector (dad) l-2455 at uv range 200–400 nm. the mobile phase consisted of solvent a: methanol / 0.5% acetic acid 1:4 (v/v) and solvent b: methanol. the gradient was as follows: (a:b) 100:0 for 0–25 min; 70:30 for 35 min; 50:50 for 45 min; 0:100 for 50–55 min; 100:0 for 57–67 min. the comparison of the uv spectra at λ = 254 and retention times with standard compounds enabled the identification of phenolic acids present in the analyzed samples. the quantitative analysis was performed using a calibration curve with the assumption of linear size of the area under the peak and the concentration of the reference standard. determination of fatty acids. one gram of powdered lyophilized biomass was extracted with chloroform/methanol solution, 2:1 (v/v). the fatty acid methyl esters (fame) were synthesized using 20% bf3 in methanol at 100°c. the fame analyzes were done with gas chromatography – agilent 6890 n with capillary column j&w db – 23 (60 m, id 0.25 mm, 0.25 µm) and fid detector. the chromatograph parameters: fid 260°c, injector 250°c, split ratio 50:1, oven 140°c for 5 min, ranged from 140 to 190°c at 4°c/min, 190°c for 15 min, and ranged from 190 to 240°c at 2.75°c/min, 240°c for 4 min, carrier gas – helium. for the identification of the fatty acids (fa), retention times of standards fame from supelco (47801) were used. the peak areas were measured with an integrator (chemstation). the results for fa composition, total saturated fatty acids (sfa), monounsaturated fatty acids (mufa), and polyunsaturated fatty acids (pufa) in the samples were expressed as relative percentage of the total fatty acids. determination of sterols. five grams of powdered lyophilized biomass was extracted with a mixture of methanol/dichloromethane 7.5:2.5 (v/v). the mixture was sonicated at 40 khz for 10 min. after 2 hours, the extract was centrifuged at 12 000 rpm for 4 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1086 sułkowska-ziaja et al. / chemical analysis of mycelial culture of sarcodon imbricatus 10 min and decantated. the extraction procedure was repeated twice and the obtained extracts were mixed together and evaporated under reduced pressure. the hplc analysis was conducted on a hitachi (merck, germany) liquid chromatograph equipped with a detector uv-vis l-7400. sterols were separated and analyzed using an rp18 column (4.6 × 250 mm, 5 μm) at 30°c. the mobile phase consisted of solvent a: methanol/water, 80:20 (v/v) and solvent b: methanol/dichloromethane 75:25 (v/v). a gradient procedure was used as follows: starting with the sample injection, 60% of solvent b for 5 min, a linear gradient from 60 to 100% of solvent b for 10 min, and 100% for 10 min. the flow rate was 1.0 ml min−1 [18]. the quantitative analysis was performed using a calibration curve with the assumption of linear size of the area under the peak and the concentration of the reference standard. the isolation and purification of ergosterol and ergosterol peroxide were performed with chromatographic methods using hp-tlc plates (merck millipore, germany). the 1h-nmr spectra were recorded using the mercury 300-brüker 1h 300.08 mhz apparatus in nmr spectroscopy laboratory of the department of organic chemistry at jagiellonian university medical college in kraków. statistical analysis all received data were presented as mean ± standard deviation (sd) and subjected to statistical analysis (statistica version 12 pl software package, statsoft). statistical significance of differences between studied groups was estimated with non-parametric mann–whitney u test. results and discussion optimization of culture conditions from among the tested carbon sources, the highest mean dry biomass 8.34 g l−1 dw was observed for the medium containing fructose. the biomass growth on the medium supplemented with glucose was also high in compare to other carbon sources (7.24 g l−1 dw) (fig. 1a). glucose and fructose are commonly used as carbon sources for in vitro cultures of higher fungi. of the six carbon sources tested by joo [19], including glucose, fructose, maltose, and sucrose, the largest mycelium growth in s. aspratus was noted on the medium contained glucose and fructose [19]. when considering the analyzed nitrogen sources, the highest mean dry biomass (7.76 g l−1 dw) was observed on the medium containing casein hydrolysate; also the growth was substantial on the medium supplemented with urea (6.43 g l−1 dw) (fig. 1b). the low biomass of s. imbricatus mycelium was noted on the medium with the addition of inorganic nitrogen sources. the studies by joo proved that peptone was the best nitrogen source for biomass growth in s. asptatus [19]. the studies conducted in other research centers have indicated the preference for using organic nitrogen sources for biomass growth in mycelial cultures over inorganic sources [20]. this was also demonstrated by newcomb and jennison [21], who tested the nitrogen requirement when it was delivered from inorganic sources (ammonium salts) vs. organic sources (amino acids, casein, and peptone) in 42 species of fig. 1 effect of carbon (a) and nitrogen (b) sources on the mycelium biomass growth of sarcodon imbicatus. different letters indicate significant differences with p < 0.05. 5 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1086 sułkowska-ziaja et al. / chemical analysis of mycelial culture of sarcodon imbricatus fungi, including three polyporales species, e.g., genera peniophora, polyporus, and trametes. these authors reported that only a single species, trametes serialis, grew in the presence of ammonium chloride as an exclusive source of nitrogen [21]. thus our results confirmed the previous data about the suitability of organic nitrogen sources in mycelial cultures of basidiomycetes. to investigate the effect of temperature has on mycelial growth we incubated stationary liquid cultures on the standard medium at temperatures ranging from 20 to 35°c. the most suitable incubation temperature for mycelial cultures was found to be 20°c. the final biomass after a 3-week incubation at 20°c was 10.96 g l−1 dw and was eight times higher than the biomass noted at 35°c (2.72 g l−1 dw) (fig. 2a). production of mycelium occurs over the wide range of temperature. numerous studies have found an optimum temperature for growths of basidiomycota between 20 and 30°c [22,23]. obtained result is comparable with those of many kinds of mushrooms that exhibit relatively lowtemperature optima, ranging from 20 to 25°c in their submerged cultures [24]. the optimum initial ph value was estimated to be 6.0. when this value was achieved, a biomass growth of 6.0 g l−1 dw was noted after the 3-weeks incubation period, which was 20 times higher than that observed on the medium with lowest ph value (3.0) amounting only to 0.28 g l−1 dw (fig. 2b). for the mycelium growth of different species of higher fungi, the optimal ph value ranges from 5.0 to 6.0, except for the arboreal species in which the optimal ph can be lowered down to 4.0 [23,25]. in our research, the highest amount of mycelium biomass of s. imbricatus was obtained at the initial medium ph = 6. moreover, further lowering (to ph = 3) or increasing (up to ph = 7.5) of the ph of culture medium results in many times smaller mycelial growth. our results confirm that the optimal ph value for axenic culture of s. imbricatus amounts around 6.0, and increase up to 7.5 results in a significant decrease in the biomass growth. many researchers consider that the acidity of the medium is critical for the biomass formation and metabolites accumulation. they suggest that a suitable ph value of the medium is crucial for normal functioning of the cellular membranes, maintenance of cell structure, and morphology, and also for the intake of nutrients and synthesis of active metabolites [26]. based on the obtained results expressed as a dry biomass increments, the medium variant containing fructose at 50 g l−1 as a carbon source and casein hydrolysate at 10 g l−1 as nitrogen source was chosen for further studies. the optimum incubation temperature was determined as 20°c and the initial ph value was set at 6.0. the best increase in biomass was observed during 3-week growth cycles in shaking liquid cultures and averaged at 10.2 g l−1 dw. this increase in biomass and the dynamics of mycelium growth differ from the results that we had obtained for xerocomus badius and tricholoma equestre cultures studied earlier [27]. obtained during the optimization of biomass s. imbricatus increases were higher than in previously conducted mycelial cultures of x. badius and t. equestre. the increments of biomass amounted to 5.2 and 7.2 g l−1 dw for x. badius and t. equestre, respectively. in all of the studied cultures were obtained homogeneous biomass in the form of spherical aggregates [27]. mycelial cultures of these species were maintained on the basic variant of a medium, without the modification of the culture condition. the obtained results indicate the need to carry out the optimization of mycelial cultures. fig. 2 effect of temperature (a) and initial ph (b) of the medium on the mycelial biomass growth of sarcodon imbicatus. different letters indicate significant differences with p < 0.05. 6 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1086 sułkowska-ziaja et al. / chemical analysis of mycelial culture of sarcodon imbricatus determination of phenolic acids among the 14 phenolic acids studied, only three of them were detected in the tested extracts (tab. 1). the total amount of free phenolic acids was 1.86 mg × 100 g−1 dw. from the results of analyzes, it was found that mycelium from in vitro cultures of s. imbricatus contained derivatives of benzoic acid: p-hydroxybenzoic, protocatechuic, and syringic acids. in quantitative terms, the main metabolite was protocatechuic acid (1.27 mg × 100 g−1 dw). the available literature contains data on the occurrence and amounts of phenolic acids in the fruiting bodies. however, up to now phenolic acids, has not been identified in the biomass from in vitro cultures of basidiomycota. the phenolic acids are the main group of phenolic compounds produced by mushrooms. their strong antioxidant activity and ability to protect important cellular structures such as cellular membranes, structural proteins, enzymes, cell membrane lipids, as well as nucleic acids against oxidative stress are the reason for their wide biological action [28]. most common phenolic acids in fungi are gallic, ferulic, gentisic, phydroxybenzoic, p-coumaric, caffeic, protocatechuic, syringic, and vanillic acids [29]. the analyzes carried out on 23 species of mushrooms growing in their natural state (india) showed that among the phenolic acids present in mushrooms, gallic, gentisic, and protocatechuic acids occurs in large quantities. the highest amount of gallic acid is observed in morchella conica (12.85 mg g−1), and protocatechuic and gentisic acids in helvella crispa (18.48 mg g−1 and 4.89 mg g−1, respectively). other phenolic acids, i.e., ferulic, caffeic, coumaric, syringic, and vanillic are present in small or trace amounts in fungi [30]. compared to edible mushrooms, medicinal mushrooms have a higher total content of phenolic compounds [31]. in the studies conducted by barros, levels of phenolic acids had been determined in more than a dozen species of fungi from the genera agaricus, lactarius, and lycoperdon commonly found in central europe [32]. the study also included the fruiting bodies of the s. imbricatus in which p-hydroxybenzoic acid was determined at 3.32 mg × 100 g−1. in other fruiting bodies of basidiomycota analyzed by barros, protocatechuic acid was found in considerable amounts, e.g., 34.2 mg × 100 g−1 in the species ramaria botrytis [32]. determinations of fatty acids the present study is the first of its kind which determined the composition of free fatty acids (ffa) in the biomass from mycelia cultures of s. imbricatus and showed that the in vitro cultures were a good source of therapeutically important ffa. the percentage of fatty acids in the biomass from mycelial cultures is shown in tab. 2. one of quantitatively predominant compounds in our studies was α-linolenic acids (cis,cis,cis-9,12,15-octadecatrienoic acid) (23.32%). this compound possesses multidirectional therapeutic action: is indispensable for normal lipid transport, participating in prostaglandin synthesis, lowering blood pressure, and changing blood coagulation ability [33]. free fatty acids are the common components of the species belonging to basidiomycota and their presence in spores is an important determinant for taxonomy [34]. ffa contents range from 20% to 30% and the substantial amount of unsaturated fatty acids, even up to 70% of the total fatty acids, is conspicuous [35]. the previous studies showed the presence of palmitic, oleic, and linoleic acid in fruiting bodies of basidiomycota. the unsaturated fatty acids mainly belonged to the ω6 and 9 family indicating the carboxyl-directed desaturation as a major metabolic pathway [36]. low fatty acid contents are characteristic of the fruiting bodies of the species agaricus bisporus and pleurotus ostreatus (0.3 and 0.486 mg × 100 g−1 dw, tab. 1 amounts (mg × 100 g−1 dw) of phenolic acids and sterols in biomass extracts from in vitro cultures of sarcodon imbricatus cultivated on the optimized medium described by turło [16], during 3-weeks growth cycles. name of compound amount phenolic acids p-hydroxybenzoic acid 0.31 ±0.002 protocatechuic acid 1.27 ±0.020 syringic acid 0.29 ±0.054 total content 1.86 ±0.040 sterols ergosterol 197.70 ±1.99 ergosterol peroxide 200.47 ±1.08 total content 398.17 ±0.99 all experimental data are mean ± standard deviation (sd) of triple determination. 7 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1086 sułkowska-ziaja et al. / chemical analysis of mycelial culture of sarcodon imbricatus respectively) [37]. however, cantharellus cibarius and boletus edulis contain up to 8–10 times greater quantities of fatty acids. lauric acid (dodecanoic acid), myristic acid (tetradecanoic acid), and palmitic acids (hexadecanoic acid) dominate among the saturated fatty acids, whereas oleic (9-cis-octadecenoic acid) and linoleic (cis,cis-9,12octadecadienoic acids) prevail among unsaturated ones. more recent data describe that the coriolinic acid (1,3-hydroxy-9-cis,11-trans-octadecadienoic acid) and linoleic acid isolated from pleurotus pulmonarius showed anthelminthic properties [38]. in the study of barros et al. [39], the major fatty acids found in s. imbricatus were linoleic and oleic acids followed by palmitic acid. this observation is in agreement with the results of our study, but we also detected a high amount of linolenic acid in the mycelial cultures which may suggest the conversion of linoleic acid into linolenic one. the sum of unsaturated fatty acids was similar to the content of these acids detected by barros [39]. our studies suggest an influence of the medium components on the synthesis of fatty acids. tab. 2 contents (%) of fatty acids in biomass extract from in vitro cultures of sarcodon imbricatus cultivated on the optimized medium described by turło [16], during 3-week growth cycles. systematic name lipid numbers % sd sfa octanoic acid c8:0 3.55 0.45 decanoic acid c10:0 0.96 0.07 dodecanoic acid c12:0 1.82 0.11 tridecanoic acid c13:0 0.94 0.05 tetradecanoic acid c14:0 0.91 0.01 pentadecanoic acid c15:0 0.73 0.04 hexadecanoic acid c16:0 18.64 0.50 heptadecanoic acid c17:0 0.32 0.02 octadecanoic acid c18:0 0.54 0.01 eicosanoic acid c20:0 31.09 0.39 docosanoic acid c22:0 0.52 0.04 mufa 9-tetradecenoic acid c14:1cis9 0.20 0.05 7-hexadecenoic acid c16:1ω7 0.66 0.51 trans-9-octadecenoic c18:1ω9 trans 0.83 0.02 cis-1-eicosanoic c20:1 0.34 0.01 9-cis-octadecenoic acid c18:1ω9 12.77 0.41 cis-13-docosenoic c22:1ω9 0.67 0.03 pufa cis,cis,cis-9,12,15-octadecatrienoic acid c18:3ω3 23.32 0.37 5,8,11,14-all-cis-eicosatetraenoic acid c20:4w6 1.19 0.08 sfa – saturated fatty acids; mufa – monounsaturated fatty acids; puf – polyunsaturated fatty acids. all experimental data are mean ± standard deviation (sd) of triple determination. 8 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(2):1086 sułkowska-ziaja et al. / chemical analysis of mycelial culture of sarcodon imbricatus determination of sterols hplc analysis of the extracts from the biomass cultured in vitro revealed peaks with retention times corresponding to the retention time of ergosterol (ergosta-5,7,22trien-3β-ol) and ergosterol peroxide (5α,8α-epidioxy-22e-ergosta-6,22-dien-3β-ol) standards. its contents were determined from a standard curve and showed that content from in vitro cultures amounted 197.70 and 200.47 86 mg × 100 g−1 dw, respectively (tab. 1). there are no data on the content of ergosterol in in vitro cultures of s. imbricatus and the present study is the first one that documented the identity of sterols in these cultures with spectral methods. the identity of ergosterol and ergosterol peroxide were confirmed by comparing the obtained spectral data with literature data [40,41]. the sterols are widespread chemical compounds of fungi. ergosterol and its peroxide occur in a majority of representatives of basidiomycota. for instance, the mean ergosterol content in the genus lactarius is 269.01–300.86 mg × 100 g−1 dw and in cantharellus 304.12–377.86 mg × 100 g−1 dw. it was proven that these compounds are essential for the normal development of hyphae of higher fungi while ergosterol is the main part of fungal cell membranes. although it occurs in a majority of basidiomycota species, the highest content of this compound was noted in saprophytic fungi [42]. the ergosterol irradiated by uv light is transformed into vitamin d2. it is also a precursor of cortisol, an adrenal cortex hormone producing anti-inflammatory actions [43]. many studies have demonstrated that ergosterol and its peroxidation products may give potential health benefits and significant pharmacological activities, including reducing pain related to inflammation, reducing the incidence of cardiovascular disease, and inhibiting cyclooxygenase enzyme (cox), antioxidant, antimicrobial, anticomplementary, and antitumor activities [44,45]. moreover, ergosterol peroxide isolated from the acetone extract of fruiting bodies of sarcodon aspratus berk. s. ito inhibits proliferation and induce apoptosis of acute lymphoblastic leukemia (hl60) cells [41]. apart from ergosterol and its peroxide, among representatives of mushrooms, attention should be paid to: brassicasterol, ergosta-5,24(28)-dienol, stigmasta-7,24(28)dienol, stigmast-7-enol, 5-dihydroergosterol, or 22-dihydroergosterol, and cholesterol and its derivatives: 24-methylene-cholesterol, 24,25-methylene-cholesterol, 24-ethyl– cholesterol, lanosterol, desmosterol, and episterol [46]. conclusions optimization of axenic culture of s. imbricatus resulted in defining the medium with the greatest efficiency of biomass growth. the chemical analyzes of obtained homogeneous mycelium demonstrated the ability to the endogenous accumulation of free phenolic acids, fatty acids, and sterols. these metabolites are characterized by a wide spectrum of biological activity. further studies are planned to optimization of mycelial cultures aimed at stimulation of metabolites production by the addition of compounds precursors. acknowledgments the authors would like to thank prof. maria rudawska and dr. tomasz leski for 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molecular targets as cancer therapeutics. appl microbiol biotechnol. 2005;67(4):453– 468. http://dx.doi.org/10.1007/s00253-004-1787-z 46. weete jd, abril m, blackwell m. phylogenetic distribution of fungal sterols. plos one. 2010;5(5):10899. http://dx.doi.org/10.1371/journal.pone.0010899 http://dx.doi.org/10.1271/bbb.69.212 http://dx.doi.org/10.1016/0079-6832(75)90002-6 http://dx.doi.org/10.1016/j.foodchem.2004.08.022 http://dx.doi.org/10.1024/0300-9831.73.1.19 http://dx.doi.org/10.1007/s00253-004-1787-z http://dx.doi.org/10.1371/journal.pone.0010899 abstract introduction material and methods mushroom material and obtaining in vitro culture optimization of culture conditions chemical analyzes statistical analysis results and discussion optimization of culture conditions determination of phenolic acids determinations of fatty acids determination of sterols conclusions acknowledgments references 2016-12-27t22:55:36+0000 piotr otręba pruning wastes from fruit trees as a substrate for pleurotus ostreatus acta mycologica article id: 568 doi: 10.5586/am.568 publication history received: 2021-01-20 accepted: 2021-04-21 published: 2021-08-27 handling editor andrzej szczepkowski; warsaw university of life sciences – sggw, poland; https://orcid.org/0000-00029778-9567 authors’ contributions cpf and aylt: research designing; cpf and yado: methodology; acsl: investigation, resources (experimental materials, chemicals, and instruments), and funding; ceg and yado: data curation; aylt: writing – original draft, supervision; ceg: writing – review and editing; cpf: project administration funding this research was supported by juan de castellanos university. competing interests no competing interests have been declared. copyright notice © the author(s) 2021. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. original research paper in mycology pruning wastes from fruit trees as a substrate for pleurotus ostreatus angela yaneth landínez-torres 1, carmenza pérez fagua 1, angie coraima sanabria lópez2, yuli alexandra deaquiz oyola 1, carolina elena girometta 3* 1faculty of agrarian and environmental sciences, juan de castellanos university, colombia 2administrative management of the municipality of san eduardo, boyacá, colombia 3department of earth and environmental sciences, università degli studi di pavia, italy * to whom correspondence should be addressed. email: carolinaelena.girometta@unipv.it abstract plant material obtained by pruning and production of deciduous fruit trees was evaluated as substrates for the production of the oyster mushroom, pleurotus ostreatus. lignified branches and stems from peach, apple, and pear trees were processed using a ripping machine to reduce the size of chips and to optimize disinfection. a completely randomized experimental design was proposed with six treatments (novel substrates) and one absolute control (100% hay substrate). morphological variables such as thickness and diameter of the pileus, stipe length were assessed, as well as production variables (sprouting, fresh weight, and biological efficiency) and bromatological analysis (ash, ethereal extract, crude fiber, and crude protein). apart from the 100%-hay substrate, biological efficiency ranged between 27% (100% apple tree as the substrate) and 140% (50% hay + 50% peach tree as the substrate). according to morphological analysis, the highest diameters were recorded from mixed substrates (50% hay + 50% wood), and a highly significant positive correlation was found between diameter and stipe length. morphological parameters were not significantly correlated with biological efficiency. analysis of biological efficiency confirmed that mixed substrates clustered together with 100%-wood substrates. bromatological analysis showed that the mixed substrate (50% hay + 50% pear tree) had the highest protein content among the novel tested substrates. bromatological parameters were not significantly correlated with biological efficiency. in conclusion, pruning residues from fruit trees can be valuable by using them as substrates for the cultivation of p. ostreatus. production is quantitatively competitive with that using hay, on the condition that wood is mixed with hay. keywords fruit trees; mushrooms; pleurotus ostreatus; sustainable production; lignocellulose residues 1. introduction macromycetes of the genus pleurotus (fr.) p. kumm. are widely grown and consumed worldwide, especially in europe and asia. pleurotus is highly nutritious, possesses medicinal characteristics, and are highly appreciated in the international cuisine for their organoleptic characteristics. according to research and markets (https://www.researchandmarkets.com/), “the mushroom cultivation market was estimated to account for a value of usd 16.7 billion in 2020.” e champignon – agaricus bisporus (j. e. lange) imbach – stands out as the species with the highest acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 1 https://doi.org/10.5586/am.568 https://orcid.org/0000-0002-9778-9567 https://orcid.org/0000-0002-9778-9567 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0002-5360-5252 https://orcid.org/0000-0002-7355-857x https://orcid.org/0000-0002-3720-8724 https://orcid.org/0000-0002-2583-4664 mailto:carolinaelena.girometta@unipv.it landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues global production (40%), followed by oyster mushrooms sensu lato (pleurotus spp.) and shiitake, lentinula edodes (berk.) pegler. e oyster mushroom p. ostreatus (jacq.) p. kumm. is one of the most studied edible mushrooms because of its nutritional quality, ease of cultivation, and economic potential (research and markets, 2021). a very rich literature has never ceased to provide information on the cultivation of this species; the main parameters and protocols were summarized by stamets and chilton (1983) and updated by several authors (mahari et al., 2020; sánchez, 2010). e cultivation of this product started in 1917–1919, but it has significantly developed in the sixties, mainly in italy, former czechoslovakia, and hungary. subsequently, it spread throughout the rest of europe both for commercialization and self-consumption. originally performed on trunks and straw, p. ostreatus cultivation has been increasingly focused on agricultural wastes and residues within a circular economy vision (grimm & wösten, 2018; masevhe et al., 2016; ozcariz-fermoselle et al., 2019). biologically, p. ostreatus is a saprotroph species naturally occurring on several broadleaves. e pileus is cream colored to lead gray, 5–20 cm, typically ear-shaped or shell-shaped. lamellae are grayish-whitish and decurrent. e stipe is eccentric, 2–3 cm long; smell and taste are pleasant and flesh is white and quite firm (boccardo et al., 2008). pleurotus ostreatus sensu stricto is regarded as worldwide distributed, although oen in sympatry with other strictly related species, e.g., p. cornucopiae (paulet) rolland and other oyster mushrooms, e.g., p. eryngii (dc.) quél. (zervakis et al., 2014). regarding its nutritional quality, protein content from p. ostreatus is compared to that of meat, since 80% of p. ostreatus protein is digestible. in addition, p. ostreatus provides minerals (p, fe, ca, k), thiamine (vitamin b), riboflavin (vitamin b2), pyridoxine (b6), pantothenic acid, biotin, folic acid, nicotinamide, ascorbic acid (vitamin c), and ergosteine (provitamin d) (bayona, 2012; la guardia et al., 2005). since it is a saprotroph, p. ostreatus plays an important ecological role in the disintegration of organic waste because of its ability to degrade cellulose – the main component of the plant cell wall – an abundant material that is difficult to decompose because of its internal structure. is trophic strategy facilitates cultivation, as materials from the wild with similar residue composition can be used as a substrate (hernández & lópez, 2006). stems and branches produced by sanitary pruning of deciduous fruit trees are considered a limitation for the production of these crops, as their inadequate management hinders activities such as weeding, phytosanitary applications, and fertilization. furthermore, unmanaged residues are a potential source of pests and diseases. edible fungi such as p. ostreatus, which are useful for degrading cellulose through the production of enzymes, can use semiwoody and lignocellulosic structures rich in ash, protein and fiber as substrates to grow. in recent years, colombia has remarkably increased the cultivation of edible mushrooms. e oyster mushroom has been regarded as the finest species to grow due to the traditional method of planting, rapid propagation, and cultivation with low requirements for technological inputs (fernandez uribe, 2014). moreover, colombia is a major producer of apples, peaches, and pears (particularly in the region of boyacá); therefore, proper handling of pruning residues from trees represents a challenge for sustainable agriculture and a chance of production diversification for small farmers. finally, biodegradation of spent mushroom compost in agroenvironments is particularly favored by colombian climatic conditions and the richness of soil microbiota (landínez-torres et al., 2020; landinez-torres et al., 2019). e aims of this study were: (i) to explore the potential of pruning residues from selected fruit trees as a substrate for the efficient cultivation of p. ostreatus, and (ii) to provide a user-friendly and scalable methodology for the cultivation of p. ostreatus by rural families. acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 2 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues 2. material and methods 2.1. study area e field work was performed in the municipality of nuevo colón-boyacá, on the faca farm located at 05°21′45″ n and 73°27′39″ w, located at 2,561 m a.s.l, characterized by an annual average temperature of 15.1 °c, annual precipitation of 920 mm, and relative humidity of 66%. cultivation of the oyster mushroom p. ostreatus was performed at the experimental farm san isidro labrador, soracá-boyacá, belonging to the juan de castellanos university. 2.2. experimental design six novel cultivation substrates were tested; hay was assumed to be the absolute control, since it is a common and well-known substrate for p. ostreatus cultivation. e treatments and absolute control consisted of four replicates. substrates were set as follows (%w/w; based on wet weight): t1 – 100% hay; t2 – 100% apple tree chips; t3 – 100% peach tree chips; t4 – 100% pear tree chips; t5 – 50% hay + 50% apple tree chips; t6 – 50% hay + 50% peach tree chips; and t7 – 50% hay + 50% pear tree chips. 2.3. field work pruning residues from apple trees, malus domestica (suckow) borkh., peach trees, prunus persica (l.) batsch, and pear trees, pyrus communis l., from faca farm and hay from local markets were mechanically milled to about 1 cm chips using gtm professional gts 1300 wood chipper (brumar srl; asti, italy). subsequently, wood chips and milled hay were separately sterilized in boiling water (approximately 80 °c due to altitude a.s.l.) for 1 hr. part of the volume of water (75%) was poured into the sterilization container. further operations were not performed axenically. 2.4. cultivation of pleurotus ostreatus clear plastic bags (40 cm × 60 cm) were used for cultivation. spawn of p. ostreatus provided by the national training service (servicio nacional de aprendizaje, sena) of duitama, boyacá (colombia) was selected as the inoculum source. substrate layers (wood chips, hay, or a mixture of both) were alternated with spawn layers into the cultivation bags. e weight of 235 g per bag was set, with the inoculum being <12.5%. cultivation bags containing the substrate and inoculum are hereaer referred to as breads. incubation was performed in a clean room at a relative humidity of 81% and an average temperature of 19.1 °c (thermo-hygrometer uni-t reference ut 333; uni-trend technology china). breads were gently covered by a black geotextile to simulate dark conditions for 15–20 days aer inoculation. when the primordia sprouted, the geotextile was removed, but direct lightening was avoided. aer primordia sprouting was observed, basidiomes were allowed to grow for 7–10 days before harvest. in parallel, a subsample of basidiomes was allowed to grow for 29 days to record the growth profile. 2.5. bromatological analysis bromatological analysis (ash, crude protein, ethereal extract, and crude fiber) was performed at the animal nutrition laboratory of the pedagogical and technological university of colombia (uptc). preliminary dehydration and measurement of dry weight were performed at the plant physiology laboratory of the juan de castellanos university foundation. acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 3 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues a b ab b ab ab b 0 2 4 6 8 10 12 t1 t2 t3 t4 t5 t6 t7 n o . o f s p o ro m e s / b a g treatments figure 1 sprouting: average number of sporomes/bag for each treatment. e analysis was performed in duplicates. basidiomes of p. ostreatus from each treatment and hay were analyzed, as were wood chips from apple trees, peach trees, and pear trees. for preliminary dehydration, samples were maintained in paper bags with the least amount of air possible, dried at a maximum temperature of 60 °c for 48 hr, and then processed in a mill with a 1-mm-diameter pore mesh. e following methods were used for the bromatological analysis of p. ostreatus and the substrates used for production: the oxidation method (helrich, 1990), crude protein with kjeldahl (association of official analytical chemists, 1984), raw fiber with acid detergent (horwitz, 2000), and ethereal extract with soxlhet (church et al., 2002). all analyses were performed according to the official laboratory protocols of the animal nutrition laboratory of uptc (universidad pedagógica y tecnológica de colombia). 2.6. data analysis for the data analysis, the fulfillment of statistical assumptions (normality, homogeneity of variance, and independence) was verified. to determine significant statistical differences between treatments, the analysis of variance (anova) was performed, followed by tukey’s multiple comparison test with a significance level of 0.05. data analysis was performed using rstudio statistical soware (r 4.0.4 coupled with rstudio 1.4.1103 interface, rstudio, boston, usa). cluster analysis, mann–whitney u test, kruskal–wallis test, dunnett’s t3 test, and spearman’s ρ test were performed by ibm spss statistics 21 (ibm, new york, usa). other graphic designs were obtained using microso excel 2016. 3. results regarding sprouting, estimated by the number of fingers obtained per bag, there were significant statistical differences confirmed by all the tests performed (mann–whitney u, kruskal–wallis, and tukey’s). as shown in figure 1, the treatments with the highest average number of sporomes were t1 (nine sporomes/bag), followed by t6 (eight sporomes/bag); t7 had the lowest number of sporomes (four sporomes/bag). sprouting was not correlated with any other morphological variables (pileus diameter, pileus thickness, and stipe length). morphometry relied on three variables: pileus diameter, pileus thickness, and stipe length. significant statistical differences, confirmed by all the tests performed (mann–whitney u, kruskal–wallis, and tukey’s), were found among treatments for all three variables. in the first week, t6 presented the greatest thickness of the pileus (0.32 cm), while t1 and t2 presented the lowest (0.10 cm). in the second week, t1 and t6 presented the greatest thickness of the pileus (0.50 cm), while t4 presented the lowest thickness of the pileus (0.32 cm). in the third week, t2 presented the highest acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 4 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues figure 2 ickness profile of pileus as a function of time. figure 3 diameter profile of pileus as a function of time. thickness of the pileus (1.50 cm), followed by t5 (1.46 cm), while t7 presented the lowest thickness of the pileus (0.40 cm) (figure 2). regarding the diameter of the pileus, in the first week, t6 presented the greatest diameter of the pileus (3.2 cm), while the lowest values were obtained by t1 (1 cm), followed by t2 (<1 cm). in the second week, t6 presented the largest diameter (8 cm), followed by t1 (7.6 cm), while t4, on the other hand, presented the smallest diameter (2.15 cm). in the third week, there were no significant differences; t2 presented a diameter of the pileus of 7.8 cm, followed by t1, at 7 cm (figure 3). regarding the length of the stipe, in the first week, t6 presented the longest stipe (3.20 cm), followed by t1 (1.68 cm), while t2 presented <1-cm stipes. in the second week, t6 treatment presented the longest stipe (3.85 cm), followed by t7 (3.56 cm); t2, on the other hand, presented the shortest length, at <1 cm. in the third week, there were no significant differences; t7 presented a stipe length of 2.12 cm, followed by t2, which presented a length of 2 cm (figure 4). according to the correlation analysis (spearman’s ρ), a significant positive correlation was found between the diameter of the pileus and stipe length (ρ = 0.707, sig. h0 = 0.0%). e morphology of sporomes grown on mixed substrates (hay + wood) was shied toward higher diameters and stipe lengths than sporomes grown on pure wood or pure hay (figure 5). acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 5 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues figure 4 stipe length profile as a function of time. 0 2 4 6 8 10 diameter thicknesslength t1 t2 t3 t4 t5 t6 t7 figure 5 synopsis of morphological parameters of sporomes in different substrates. regarding fresh weight, according to statistical analysis, t6 presented the highest fresh weight per sporome (27.2 g), followed by t1 (19.2 g), and t5 (16.25 g) (figure 6). although the highest fresh weight values were found in t6 and t1, as well as the highest values of sporomes per bag, there was no significant correlation between average fresh weight per sporome and number of sporomes per bag. biological efficiency (b.e.%), which is the % ratio between the cumulative fresh weight of mushrooms and dry weight of the substrate (ahmed et al., 2009; chang et al., 1981), was the highest in hay (figure 7). nevertheless, it should be noted that dry hay is a considerably lighter matter than wood. us, this result must not mask the values of b.e.% on other substrates, with particular concern to t6 (140%) and t5 (76%). more interestingly, in figure 8, the ratio between b.e.% in hay and other examined substrates (“normalized b.e.%” hereaer) is shown. e highest normalized b.e.% was verified in t6; as a whole, mixed substrates recorded higher values than those of pure wood. cluster analysis also highlighted this result (figure 9). bromatological analysis (table 1) revealed similar comprehensive profiles in sporomes from different substrates according to kruskal–wallis test (p = 0.996). e bromatological profiles of pure substrates (hay and wood from different trees) were similar to each other (p = 0.962) as well. as expected, fungal profiles significantly differed from wood profiles, as they concern all the parameters: ash acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 6 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues 0 5 10 15 20 t1 t2 t3 t5 t6 t7 av e ra g e f re sh w e ig h t/ sp o ro m e ( g ) treatments figure 6 average fresh weight (g) of sporomes grown on different substrates. 0 50 100 150 200 250 300 350 t1 t2 t3 t5 t6 t7 b .e .% treatments figure 7 biological efficiency (b.e.%) of production on different substrates. 0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45 0.5 t2 t3 t5 t6 t7 n o r m a li z e d b .e .% treatments figure 8 normalized biological efficiency. acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 7 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues figure 9 cluster analysis of normalized b.e.% by ibm spss 21. clustering method: between groups linkage; measure by squared euclidean distance. each substrate was treated as a variable. table 1 bromatological analysis of pleurotus ostreatus from different substrates and analysis of pure wood. ashes (%) crude protein (%) ethereal extract (%) crude fiber (%) sporomes t1 8.2 16.9 6.7 18.4 t2 7.9 13.5 5.5 17.2 t3 7.2 16.0 5.2 20.1 t5 8.2 15.8 3.3 17.9 t6 8.6 13.4 3.1 19.4 t7 8.4 16.6 4.6 20.3 substrates hay 13.0 3.5 1.1 44.2 peach wood 12.4 3.9 1.8 42.4 apple wood 10.8 3.3 1.3 40.8 pear wood 11.3 2.9 1.5 46.5 (p = 2.0%), crude protein (p = 2.0%), ethereal extract (p = 2.0%), and crude fiber (p = 2.0%). is can be easily visualized in figure 10. moreover, figure 10 shows that wood profiles dramatically shied towards high values of crude fiber; ashes were also higher in fungal sporomes than the other treatments. on the other hand, fungi showed an almost symmetric frame, which highlights the remarkable protein content. regarding the bromatological analysis of p. ostreatus, t1 presented the highest value in terms of crude protein (16.9%), followed by t7 (16.6%), and t2 and t6 (13.5% and 13.4%, respectively). e highest crude fiber value for p. ostreatus was found in treatments t7 (20.3%) and t3 (20.1%). regarding the percentage of ethereal extract, it was observed that the treatments t1 (6.7%) and t2 (5.5%) stood out, presenting the highest levels. e highest concentration of ashes in the mushrooms was obtained in treatments t6 (8.6%) and t7 (8.4%). acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 8 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues 0 10 20 30 40 50 % ashes % crude protein % ethereal extract % crude fibre t1 t2 t3 t5 t6 t7 hay peach wood apple wood pear wood figure 10 synopsis of different bromatological profiles in sporomes (t1–t7) and substrates. 0 5 10 15 20 25 30 % ashes % crude protein % ethereal extract % crude fibre% s ta n d a rd d e v ia ti o n o n t h e a v e ra g e sporomes substrates figure 11 percentage standard deviation of the means of bromatological variables in sporomes (among treatments) and substrates. as a whole, each bromatological variable (ash, crude protein, ethereal extract, and crude fiber) showed no negligible intra-group variability (figure 11). us, beyond the comprehensive bromatological profile, ethereal extract recorded the highest % standard deviation of the means in both sporomes and substrates. regarding crude protein, the high % standard deviation of the means was due to the low values, which amplified the variation. 4. discussion and conclusions e sprouting results (number of sporomes per bag) can be attributed to the fact that the highest production of fungi is obtained in substrates rich in fiber and structural carbohydrates (savón et al., 2007). despite the fact that the fiber values were lower in hay than in other substrates examined in this study, the fiber of this material has a great ease of assimilation due to its physical constitution. in addition, hay is a well-known substrate for the cultivation of p. ostreatus (lópez-rodríguez et al., 2008). e morphometric results are consistent with the literature. as explained by benavides (2013) and lópez-rodríguez et al. (2008), the diameter of the pileus and stipe length depend on the maturity reached, as well as on the similarity of the size of the mushrooms. according to gaitán-hernández et al. (2009), the diameter of the pileus, in particular, is more related to the genetic characteristics of the mushroom than to the acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 9 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues type of substrate, since maintaining optimal environmental conditions for cultivation, the mushrooms will present homogeneity. interestingly, mixed substrates (hay + wood) favored diametric and stipe growth in comparison to pure wood or pure hay. b.e.% was similar to that reported in most studies (familoni et al., 2018; masevhe et al., 2016; tekeste et al., 2020; tisdale et al., 2006; zervakis et al., 2013). consistent with the literature, substrates composed of pure wood were less efficient than those composed of mixed hay-wood. nevertheless, the present study reported a similar or higher b.e.% than that found by jafarpour et al. (2010), who supplemented chips with more energetic materials than hay. interestingly, in this study, b.e.% was higher than that reported by siwulski et al. (2012), although the cultivation set was not with controlled lighting. a part of the literature reports slightly higher values for crude protein than those found in the present study (alam et al., 2008; oyetayo & ariyo, 2013; patil et al., 2010). nevertheless, it should be considered that quantitative analysis of total proteins is significantly affected by the method (conklin-brittain et al., 1999; mæhre et al., 2018). is is also true for values obtained from substrates as well (familoni et al., 2018). interestingly, wang et al. (2001) reported a very high % of crude protein but poor b.e.% on spent beer grains. analogously, the top crude fiber values (approximately 20%) from the present study are consistent with the literature (alam et al., 2008; oyetayo & ariyo, 2013) or remarkably higher than those (patil et al., 2010). ethereal extract is a difficult variable to compare with data reported in the literature, since it strongly depends on the method used (alam et al., 2008; oyetayo & ariyo, 2013; patil et al., 2010). in this study, ethereal extract and crude protein appeared as pivotal variables in the bromatological profile of p. ostreatus, since their variation was remarkable. unexpectedly, there was no significant correlation between the bromatological values of the substrate and sporomes. is is consistent with the results reported by vega & franco (2013), who argue that sporome development is substantially independent from the substrate when different nutrients are available, regardless of their proportion. according to baena gonzález (2005), ashes in p. ostreatus are strain-correlated instead of substrate-correlated. on the other hand, benavides et al. (2015) report a correlation between oil palm rachis in the substrate and ethereal extract of p. ostreatus sporomes. conversely, according to savón et al. (2007), substrate composition influences the quality and protein quantity of fungi, whereas ashes represent the mineral content that becomes available for fungal growth (benavides, 2013). bromatological profiles of wood substrates are consistent with the literature (familoni et al., 2018), although wingching-jones and retana (2009) found lower content of crude fiber, and jafarpour et al. (2010) found remarkably higher content of crude fiber than in other substrates. high fiber content is highly desirable; according to hoyos et al. (2012), it is essential for the growth of p. ostreatus. moreover, fibers can be incorporated into the soil as easily assimilated aggregates for crops. in conclusion, pruning residues from apple trees, peach trees, and pear trees can be successfully used as raw materials to obtain substrates for p. ostreatus cultivation. substrates composed of wood chips mixed with hay showed the best performance in terms of biological efficiency and morphometric parameters of sporomes. e 50% hay + 50% peach wood mix resulted in the highest values of those variables. moreover, sprouting of primordia took a shorter time on this substrate than on the other substrates. erefore, the methodology applied in the present study is adequate for the production of p. ostreatus. e cultivation protocol was confirmed to be user-friendly for scalable production by small farmers. substrate disinfection is a fundamental factor in the reduction of microbial competitors; moreover, heat disinfection partially alters substrate consistency and favors fungal colonization. acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 10 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues acknowledgments e authors thank the agricultural engineer, hectór osvaldo rodríguez páez, owner of the faca farm, located in the municipality of nuevo colón (boyacá); the faculty of agrarian and environmental sciences; and the juan de castellanos university, for their support and collaboration in the development of the research. references ahmed, s. a., kadam, j. a., mane, v. p., patil, s. s., & baig, m. m. v. 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(2019). promoting circular economy through sustainable agriculture in hidalgo: recycling of agro-industrial waste for production of high nutritional native mushrooms. in p. castro, a. azul, w. leal filho, & u. azeiteiro (eds.), climate change-resilient agriculture and agroforestry (pp. 455–469). springer. https://doi.org/10.1007/978-3-319-75004-0_26 patil, s. s., ahmed, s. a., telang, s. m., & baig, m. m. v. (2010). e nutritional value of pleurotus ostreatus (jacq.: fr.) kumm cultivated on different lignocellulosic agrowastes. innovative romanian food biotechnology, 7, 66–76. research and markets. (2021). retrieved july 31, 2021, from https://www.researchandmarkets.com/search.asp?q=pleurotus+ostreatus sánchez, c. (2010). cultivation of pleurotus ostreatus and other edible mushrooms. applied microbiology and biotechnology, 85(5), 1321–1337. https://doi.org/10.1007/s00253-009-2343-7 savón, r. c. b., oduardo, n. g., & lópez, a. m. (2007). fermentación sólida para la producción de pleurotus sp. sobre mezclas de pulpa de café y viruta de cedro [solid acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 12 https://doi.org/10.1007/s00253-018-9226-8 http://hdl.handle.net/10554/8275 https://doi.org/10.1021/jf0307696 https://doi.org/10.1038/s41598-019-46485-1 https://doi.org/10.3390/ijerph17228311 https://doi.org/10.1016/j.jhazmat.2020.123156 https://doi.org/10.1080/02571862.2015.1079932 https://doi.org/10.3390/foods7010005 https://doi.org/10.12816/0001537 https://doi.org/10.1007/978-3-319-75004-0_26 https://www.researchandmarkets.com/search.asp?q=pleurotus+ostreatus https://doi.org/10.1007/s00253-009-2343-7 landínez-torres et al. / cultivation of pleurotus ostreatus on pruning residues fermentation for the production of pleurotus sp. on cofee pulp mixed with cedar chips]. tecnología química, 27(2), 55–62. siwulski, m., ziombra, m., & sobieralski, k. (2012). impact of light on yielding of some pleurotus sp. strains. acta mycologica, 47(1), 65–73. https://doi.org/10.5586/am.2012.008 stamets, p., & chilton, j. s. (1983). e mushroom cultivator. first washington. tekeste, n., dessie, k., taddesse, k., & ebrahim, a. (2020). evaluation of different substrates for yield and yield attributes of oyster mushroom (pleurotus ostreatus) in crop-livestock farming system of northern ethiopia. e open agriculture journal, 14, 30–35. https://doi.org/10.2174/1874331502014010030 tisdale, t. e., miyasaka, s. c., & hemmes, d. e. (2006). cultivation of the oyster mushroom (pleurotus ostreatus) on wood substrates in hawaii. world journal of microbiology and biotechnology, 22(3), 201–206. https://doi.org/10.1007/s11274-005-9020-5 vega, a., & franco, h. (2013). productividad y calidad de los cuerpos fructíferos de los hongos comestibles pleurotus pulmonarius rn2 y p. djamor rn81 y rn82 cultivados sobre sustratos lignocelulósicos [productivity and quality assessment of fruiting bodies of the edible mushrooms pleurotus pulmonarius rn2 and p. djamor rn81 and rn82 cultivated on lignocellulosic substrates]. información tecnológica, 24(1), 69–78. https://doi.org/gpfm wang, d., sakoda, a., & suzuki, m. (2001). biological efficiency and nutritional value of pleurotus ostreatus cultivated on spent beer grain. bioresource technology, 78(3), 293–300. https://doi.org/10.1016/s0960-8524(01)00002-5 wingching-jones, r., & retana, g. a. (2009). valor nutricional del heno de transvala inoculado con el hongo pleurotus ostreatus sp. [nutritional value of transvala hay inoculated with the fungus pleurotus ostreatus]. agronomía costarricense, 33(1), 147–153. zervakis, g. i., koutrotsios, g., & katsaris, p. (2013). composted versus raw olive mill waste as substrates for the production of medicinal mushrooms: an assessment of selected cultivation and quality parameters. biomed research international, 2013, article 546830. https://doi.org/10.1155/2013/546830 zervakis, g. i., ntougias, s., gargano, m. l., besi, m. i., polemis, e., typas, m. a., & venturella, g. (2014). a reappraisal of the pleurotus eryngii complex – new species and taxonomic combinations based on the application of a polyphasic approach, and an identification key to pleurotus taxa associated with apiaceae plants. fungal biology, 118(9–10), 814–834. https://doi.org/10.1016/j.funbio.2014.07.001 acta mycologica / 2021 / volume 56 / article 568 publisher: polish botanical society 13 https://doi.org/10.5586/am.2012.008 https://doi.org/10.2174/1874331502014010030 https://doi.org/10.1007/s11274-005-9020-5 https://doi.org/gpfm https://doi.org/10.1016/s0960-8524(01)00002-5 https://doi.org/10.1155/2013/546830 https://doi.org/10.1016/j.funbio.2014.07.001 pruning wastes from fruit trees as a substrate for pleurotus ostreatus 1 introduction 2 material and methods 2.1 study area 2.2 experimental design 2.3 field work 2.4 cultivation of pleurotus ostreatus 2.5 bromatological analysis figure 1 2.6 data analysis 3 results figure 2 figure 3 figure 4 figure 5 figure 6 figure 7 figure 8 figure 9 table 1 figure 10 figure 11 4 discussion and conclusions acknowledgments references type of manuscript: original research title: biodiversity of astraeus asiaticus, a wild indigenous edible mushroom, in the forests of bankura district, west bengal and its antioxidant property koushik pandey1, swapan kumar ghosh1 1 molecular mycopathology lab., biocontrol and cancer research unit, pg department of botany, ramakrishna mission vivekananda centenary college (autonomous), rahara, kolkata 700118, wb, india abstract three forests, beliatore, gangajal-ghati (g-ghati), and joyur, were surveyed for mushroom collection and biodiversity. mushrooms in the rhizospheric zone of some trees, such as shorea robusta, petrocarpus marsupium, terminalia bellrica, and madhuca indica, were identified at the molecular level as asterius asiaticus. thereafter, the ecological diversity of this mushroom was determined in the forests. the diversity indices of shorea robusta in the beliatore, joypur, and gangajal-ghati forests were 2.303, 2.178, and 2.36, respectively. notably, the diversity index of madhuca indica in the beliatore and joypur forests was nearly the same as that in the gangajal-ghati forest, with a value of 2.29. the total phenolics contents of the hot water, acetone, and hexane extracts of this mushroom were 6.8 ± 0.15, 3.95 ± 0.15, and 2.16 ± 0.26 mg gae/g, respectively, and the total flavonoid contents were 2.03 ± 0.12, 1.65 ± 0.2, and 1.01 ± 0.08 mg qe/g, respectively. the ascorbic acid contents in the hot water, acetone, and hexane extracts were low. the antioxidant activity of the extracts was determined using the dpph (1,1-diphenyl-2-picrylhydrazyl) assay. the ic50 values of the hot water, acetone, and hexane extracts were 42.54±1.25c µg/ml, 54.06±1.50b µg/ml, and 82.97±1.58a µg/ml respectively while that of the synthetic antioxidant, bha (butylated hydroxyanisole), was 32.41±1.26d µg/ml. overall, the hot water extract of this mushroom had the highest antioxidant content and displayed the best radical scavenging power. keywords mushroom; biodiversity; pcr; antioxidant; astraeus asiaticus running title: astraeus asiaticus mushroomits biodiversity and antioxidant property po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 1 1. introduction mushrooms, owing to their medicinal and food values, have been historically viewed as an amicable part of human society and have at times superseded plant and animal-procured products. mushrooms are recognized as functional foods with nutritional value, structural quality, and acceptability (greeshma et al., 2018; mwangi et al., 2022; pinto et al., 2020). moreover, mushrooms undergo remarkable adjustments on imminent days for adaptation as smooth-running therapeutic foods. the consumption of mushrooms in asian countries, such as india and japan, is increasing at a significant rate. based on data from research and markets (2021) (https://www.research and markets. com/), the value of the mushroom cultivation market was 16.7 billion $ (usd) in 2020, and is projected to reach 20.4 billion $ (usd) by 2025 (landínez-torres et al., 2021). palatable mushrooms are low-calorie, low-fat food adjuncts with moderate amounts of proteins, carbohydrates, vitamins, minerals, amino acids, and dietary fiber (johnsy et al., 2011). mushrooms also exhibit antibacterial, antifungal, antiviral, antiparasitic, antioxidant, anti-inflammatory, antiproliferative, anticancer, anti-hiv, hypocholesterolemic, antidiabetic, and anticoagulant activities (ghosh et al., 2020a; ghosh et al., 2020b; manimaran et al., 2021). recently, singh and varshney (2020) estimated the food value of astraeus hygrometricus, which was found to contain protein (16.02%), total carbohydrates (55.76%), fat (3.5%), monounsaturated oleic acid (4.59%), ash (3.8%), energy (159.5 kcal), pro-vitamin d2 (ergosterol) (1.09 mg/g), and dietary fiber (39.78%). among the total free amino acids (8.20 mg/g), 7 essential amino acids (3.9 mg/g), including tyrosine (0.98) and leucine (0.92), were found. these researchers also found a high amount of selenium in a. hygrometricus. in this work, we opted to focus on the mushroom, astraeus sp., owing to its food values of astraeus sp. and its prominent use as a restorative drug owing to its antidiabetic, anticancer, hepatoprotective, anti-inflammatory, and cardioprotective activities (phadannok et al., 2020). the primary aim of this study was to reveal the biodiversity of astraeus asiaticus via morphological, biochemical, and molecular research in different woodlands or forests of the bankura district, a red lateritic zone in west bengal. mycorrhizae provide nutrients between the roots of the host plant and fungi in the soil (malajczuk et al., 1982). biodiversity is a very important "natural capital." different varieties, species, genera, and their complex interactions enable a functional ecosystem and productive economies (world bank, 2022). although wild mushrooms and fungi are an integral part of a given ecosystem, a study of their diversity and type has not been performed, particularly in tropical countries, such as india. as a result, a knowledge gap exists. wild edible mushrooms act as natural resources for rural people and provide nutrition (meena et al., 2020). accordingly, an investigation of the biodiversity of wild edible mushroom and the consumption of this remarkable natural resource is of great economic importance. astraeus species po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 2 are economically important as they are collected by people for consumption and sale in commercial markets (manna & roy, 2014). bankura is recognized for its great abundance as residents can earn income by selling this wild edible mushroom. thus, this mushroom is of great socioeconomic importance in the rural areas of west bengal in india. a. asiaticus is a promising edible mushroom; however, its therapeutic value has not been explored. therefore, in this study, the molecular characteristics, biodiversity, mycochemistry, antioxidant content, and antioxidant activity of this mushroom were determined to enable future research on its medicinal importance. 2. material and methods 2.1. survey of habitat and sample collection a survey on mushrooms was conducted in the beliatore, joypur, and gangajal-ghati forest areas in bankura. joypur and beliatore in the bankura district are located in the southwest part of west bengal and is associated with native extremities owing to the damodar and dwarakeswar rivers, which indicate the depletion of both forest range areas. the damodar and dwarakeswar rivers are almost parallel toward the southeast, as the general slope of the district runs from northwest to southeast. a periodic survey was conducted in the preferred area for the collection of a. asiaticus during the rainy season from mid-july to august 2019. the habitat of mushrooms from the collection zone was noted, and photographs were captured. soil samples from a rooting depth of 30 cm were collected from 15 zones (60 quadrats) of three forests. the samples were transported to the laboratory, and processed for soil chemical parameter analysis following the standard method (iwabuchi et al, 1994, das, 2021). 2.2. identification of the mushroom species mushroom (basidioma) morphology was examined macroscopically and its size, shape, texture, color, and habit were determined, according to the pioneering work of phosri et al. (2007). biochemical tests were performed by dispensing a drop of chemicals (3% aqueous solution, 10% solution of potassium hydroxide (koh), 10% solution of feso4, and vanillin) independently and directly on the tissue of freshly collected mushrooms, and observing the color changes immediately and after 4–5 min (khaund & joshi, 2014; petrini et al., 2009). po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 3 the mushroom basidioma anatomy was observed under a compound microscope (model no. olympus cx31) after sectioning and staining with lactophenol cotton blue. images were captured using a camera attached to a microscope. for scanning electron microscopy (sem), the samples were transferred to a petri dish and immersed in 10 ml of water to rehydrate the tissues around the spores. thereafter, the spore samples were incubated in the following ethanol series with 2% glutaraldehyde for 15 min each: 30, 50, 70, 80, 90, 95, and 100%. the samples were centrifuged, and the supernatant was collected. the sample was then placed in a critical point dryer (cpd) and mounted. finally, spores were prepared for sem observation. the samples were coated with gold using a sputter coater. a focused image of the sample was used as the starting point under a scanning electron microscope. the magnification of the image was increased to a level close to the maximum, and the image was re-focused. a high-resolution sem image was captured using an attached camera. all data or characters from macroscopic, microscopic, and biochemical reactions were compared with standard keys of mushrooms (jordon, 1999; pacioni, 1981; phsori, 2007; purkayastha & chandra, 1985), and phenotypic mushroom identification was conducted. for molecular identification of mushroom species, dna was isolated from fresh basidioma using the modified hexadecyltrimethylammonium bromide (ctab) method (doyle & doyle, 1987). in brief, 100 mg sample was frozen in liquid nitrogen and ground using 500 µl lysis buffer [ctab 3% (ph 7.5), 100 mm tris hcl, 20 mm edta (ph 7.5) 1.4 m nacl2, 0.2% polyvinyl pyrolodine] and micro pestle in eppendorf tubes. the sample was then incubated at 64-65 ºc for 1 h in a water bath and vortexed at 10-min intervals. β-mercaptoethanol (20 µg/ml) was added to the tube, which was then incubated at 37 °c for 45 min. the sample was combined and centrifuged at 8000 rpm for 10 min at 4 °c using a cooling centrifuge. a total of 500 µl of chloroform: isoamyl alcohol (24:1) was added to the solution, which was vortexed under chilled conditions. the solution was centrifuged at 12000 rpm for 10 min at 4 ºc in a cooling centrifuge. the supernatant was collected in fresh eppendorf tubes. a total of 500 µl of ice-chilled isopropyl alcohol (double volume) and 3 m sodium acetate (1/10th volume) were added to the solution. cold centrifugation was then performed at 12,000 rpm for 5 min at 4 °c. the supernatant was discarded, and the pellet was collected (containing dna), dehydrated with 70% ethanol, and left to dry under laminar airflow. thereafter, the pellet was resuspended in 50-100 µl of te buffer, and stored at 4 ºc overnight. isolated dna was purified using a himedia purification kit. the purified dna was electrophoresed via electrophoresis using 0.8% agarose gel at 100 volts for 2-3 h. dna bands were observed using a uv transilluminator. po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 4 the its1–5.8sits2 marker zone for the rrna of isolated gdna was amplified using a pcr (polymerase chain reaction) protocol modified from gardes & bruns (1993), with primers its-1f (5′cttggtcatttagaggaagtaa-3′) and its4 (5′ tcctccgct tattgatatgc3′). in brief, pcr was conducted in a 50-μl reaction volume, which contained 1 ng/μl dna template, approximately 1 μl; sterile distilled water, 38 μl; 10× taq polymerase buffer with mgcl, 25 μl; 10 mm dntp mixes, 3 μl; template dna, 1 μl; its1 forward primer, 1 μl; and taq dna polymerase, 1 μl. the following pcr cycling program was employed: initial denaturation at 96 °c for 2 min followed by 35 cycles of denaturation at 96 °c for 1 min, annealing at 55 °c for 1 min, and extension at 72 °c for 2 min. the final extension at 72 °c for 10 min was terminated at the end of the amplification. the pcr product was sent to the sci genome laboratory, kerala, for sequencing. the obtained sequence was compared with other related sequences using a blast search in genbank (ncbi) (ghosh et al., 2022; podder & ghosh, 2019) and submitted to genbank, ncbi (baltimore, usa). 2.3. biodiversity analysis and mushroom extraction in different solvents biodiversity analysis fifteen study zones were selected from three forests in the bankura district. thus, 60 quadrats were used in this study. for mushroom biodiversity, quadrats (50 m × 50 m) were randomly selected and the mushroom diversity was determined. mushroom diversity was analyzed using the shannon weaver biodiversity index (h) (shannon and weaver 1962): ĥ= -∑(ni /n) loge (ni /n), where ni is the individual number of species i and n is the sum of ni. mushroom extraction in different solvents mushroom extraction was carried out using the most polar solvent (hot water), semipolar solvent (acetone), and nonpolar solvent (hexane). the collected basidiomata were washed, dried, and crushed into powder using a grinder machine. thereafter, 10 g of powder was poured separately into hot water, acetone, and hexane in a 1:10 ratio. for the hot water extract (hwe), a mixture of mushroom powder and water was boiled at 100 °c for 2 h in a water bath (chen et al., 2016). however, for the acetone and hexane extracts, the mixture of mushroom powder and solvent was shaken on a shaker at 150 rpm for 1 h and then it was placed in an incubator which was set to 38 °c for 72 h. each extract was filtered through whatman no. 4 filter paper and whatman no.1 filter paper. the filtrates were centrifuged at po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 5 3000 rpm for 15 min, concentrated through evaporation using a rotary evaporator at 50 °c, and lyophilized into powder using a lyophilizer (figure 1). the extracts were stored at −20 °c for further use. 2.4. qualitative tests of different solvent extracts of mushroom fruit body for mycochemicals flavonoid briefly, 0.2 g of extract was dissolved in 4 ml of distilled water in a test tube. thereafter, diluted sodium hydroxide (1 m) was added to the mixture, resulting in a yellow solution that turned colorless following the addition of a few drops of dilute acid (10% hydrochloric acid), confirming the presence of flavonoids (hossain et al., 2013). terpenoids approximately 0.2 g of extract was dissolved in a mixture of 2 ml of chloroform and 3 ml of concentrated sulfuric acid. a reddish-brown color eventually appeared run, indicating the presence of terpenoids (mujeeb et al., 2014). cardiac glycosides briefly, 0.2 g of the extract was combined with 2 ml of glacial acetic acid and one drop of fecl3. this mixture was then added to 1 ml concentrated h2so4. when the upper layer and the interface between the two layers turned bluish-green and reddish-brown, respectively, this indicated the presence of cardiac glycosides (auwal et al., 2014). steroids the crude extract (0.2 g) was treated with a few drops of acetic acid anhydride in a test tube. concentrated h2so4 was carefully added dropwise using the inner wall of the test tube. the presence of a brown ring on the surrounding surface indicated the presence of steroids (gul et al., 2017) saponins the extract (0.2 g) was mixed with 5 ml distilled water in a test tube. the solution was shaken vigorously for approximately 5 min, and then incubated for 30 min. the formation of a honey comb froth indicated the presence of saponin (auwal et al., 2014). po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 6 alkaloid briefly, 0.2 g crude extract was weighed in a separate test tube and warmed in 2% sulfuric acid for 2 min. in a separate test tube, the solution was mixed with a few drops of dragendroff reagent. the presence of an orange red precipitate indicated the presence of alkaloid (auwal et al., 2014). carbohydrate the extract (0.2 g) was dissolved in water (2 ml). thereafter, 2-3 drops of molisch's reagent [3.75 g of α-naphthol in 25 ml ethanol (99%)] were added to the solution for the formation of a layer without shaking the test tube. thereafter, a drop of concentrated sulfuric acid (99%) was added to the mixture. the presence of a purple color, which indicates the presence of carbohydrates, was determined at the interface (auwal et al., 2014). phlobatannins briefly, 0.2 g extract was dissolved in 2 ml of water and then added to 3 ml of dilute hcl (10%). a red precipitate indicated the presence of phlobatannins (auwal et al., 2014). phenol briefly, 0.2 g of extract was dissolved in 2 ml of distilled water. the solution was then treated with two drops of 5% fecl3. the production of a deep blue color indicated the presence of phenol (talukdar et al., 2017). tannin the extract (0.2 g) was dissolved in 4 ml of distilled water, heated in a water bath (50 °c), and filtered. two drops of 1% fecl3 were added to the filtrate. the formation of a dark green solution indicated the presence of tannins (fransina et al., 2019; usman et al., 2009). 2.5. divergent biochemical assay for antioxidant content and antioxidant potentiality antioxidant content total phenolics content assay the total phenolic content (tpc) of various solvent extracts was measured using the folin–ciocalteu reagent method (mc donald et al., 2001). briefly, 0.1 ml folin-reagent ciocalteu's (0.5 n) reagent and 0.5 ml extract were mixed and incubated for 15 min at room temperature. saturated sodium po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 7 carbonate solution (2.5 ml) was then added to the mixture, which was then incubated at room temperature for 30 min. the absorbance was measured at 760 nm. the results are expressed as mg of quercetin equivalent (qes) per g of mushroom dry weight and are presented as mean ± sd (standard deviation) of triplicate experiments. total flavonoid content (tfc) assay the tfc of the extract was evaluated using a standard guideline suggested by meda et al. (2005). briefly, equal amounts of 2% alcl3 (prepared in absolute methanol) and the extracts (1 mg/ml) were incubated in a glass tube at room temperature for 10 min. thereafter, the absorbance was measured at 415 nm. the results are expressed as mg of quercetin equivalent (qes) per g of mushroom dry weight and are presented as mean ± sd of triplicate experiments. total ascorbic acid content assay the total ascorbic acid content (taac) was determined using the folin-ciocalteu reagent method proposed by jagota and dani (1982). briefly, 0.5 ml extract solution (1 g extract in 1 ml ethyl acetate) was quickly mixed with 0.8 ml of 10% trichloroacetic acid and refrigerated for 5 min before centrifugation at 3000 rpm for 5 min. thereafter, 0.2 ml was retrieved from the mixture and mixed with 1.8 ml distilled water to a final volume of 2 ml. finally, 0.2 ml of diluted folin-ciocalteu (1:9) was added to the diluted extract mixture. after a 10 min incubation at room temperature (27 °c), the absorbance at 760 nm was measured. finally, the results are expressed as milligrams of ascorbic acid equivalents (aaes) per gram of mushroom extract, with the mean standard deviation (sd) of three replicates. antioxidant activity assay dpph assay 1,1-diphenyl-2-picrylhydrazyl (dpph) radical scavenging activity (rsa) was determined using a modified method described by bondet et al. (1997). ten different concentrations (10-100 μg/ml) were prepared from the mushroom extract, and 900 μl dpph (0.1 m) was added to each. the dpph solution was prepared by adding 2 mg dpph powder to 50 ml ethanol (0.1 m). after 2 h of incubation in the dark, the sample was subjected to a spectrophotometric study at a wavelength of 517 nm, and the absorbance was recorded. the percentage of dpph reduction was calculated using the following formula: % dpph reduction or % of inhibition = ac−as ac 𝑥𝑥 100 where ac is the absorbance of the solvent (control) and as is the absorbance of the sample. po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 8 finally, the ic50 (50% of inhibition) value was calculated. 3 results 3.1 habitat study and soil analysis the collected mushrooms were rhizospheric and ectomycorrhizal under the roots of shorea robusta, modhuca indica, petrocarpus marsupium, and terminalia bellrica. when immature, the mushrooms were subterranean but when they were fully matured, they exposed by breaking the soil surface. the soil ph of the beliatore, gangal-ghati (g-ghati) forest was 6.3, and that of the joypur forests was 6.5, 6.3, and 6.2, respectively. the soil analysis revealed that the organic carbon content of the collected soils ranged from 0.82 to 0.93%. the carbon content of beliatore soil samples was 0.93%, while that of the g-ghati soil sample was 0.87%, and that of the joypur soil sample was 0.82%. the electrical conductivity (ec) of the three forest soil samples ranged from 81.58 to 86.72 µs. the value was 86.72 µs in the beliatore forest, 84.29 µs in the g-ghati forest, and 81.58 µs in the joypur forest. the available nitrogen content of the beliatore forest sample was 220 kg/ha, while that of g-ghati, and joypur forest sample was 140 kg/ha and 180 kg/ha, respectively. the available phosphorus content in the forest soils of the bankura district ranged from 72.67 to 74.34 kg/ha, with values of 74.34 kg/ha, 73.54 kg/ha, and 72.67 kg/ha in the beliatore, g-ghati, and joypur forests, respectively. in the study area, available potassium ranged from 215.7 to 218.4 kg/ha, indicating a high potassium content in the three-forest soil sample of bankura district. the available potassium contents in the three forests (beliatore, g-ghati and joypur) were 215.7, 214.2, and 218.4 kg/ha respectively and the n:p: k ratios in these forests were 3:1:3, 2:1:3, and 2:1:3, respectively (table 1). 3.2. identification of mushroom phenotypical identification the fruiting season for this mushroom is may-august. the basidioma of this species was globose and smooth, with an unexpanded diameter of 10-16 x 8-14 mm and expanded diameter of 14-23 x 8-12 mm. the gleba was initially white; however, when the spores matured, the gleba and its granulate outer peridium turned purplish chestnut. the exoperidium was slightly viscous and smooth, and the po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 9 https://en.wikipedia.org/wiki/gleba https://en.wikipedia.org/wiki/spore https://en.wikipedia.org/wiki/gleba color was white, covering the endoperidium (figure 2). the endoperidium is a thin membranous layer of tissue, enclosing the spore mass in a 20–30 mm diameter sac, nearly round, with an irregularly shaped ostiole at the top, surface felty rough, whitish, and gray to brown color. basidiospore mass was whitish when young, and black and powdery at maturity. the spores were reddish-brown, roughly spherical with minute warts, measuring 7.5–11 μm in diameter. the mycelial layer consisted of branched hyphae that were 4–7 μm in diameter. the hyphae of the fibrous layer were 6–9 μm in diameter and branched, and the collenchyma-type layer contained branched hyphae 3–4 μm in diameter (figure 3). scanning electron micrographs provided a clear three-dimensional image and stereoscopic view of the mushroom spore and hyphal structures. ornamentation on the spore surface and hyphal structure were clearly observed (figure 4). the basidiospores showed extensive, dense, and rounded spine, and were globose, ranging from 8.65–15.3 mm diameter. other features included ornamentation, purplish chestnut color, very densely ornamented with rounded, narrow, tapered, occasionally coalescent, spines 0.9–1.50 mm long. in the present biochemical identification study, no color change was observed when 10% koh, 4% hno3, phenol, vanillin, and 10% h2so4 were employed (table 2). the color change profiles generated for each species of wild edible mushrooms in the present study served as an additional tool to identify similar mushrooms. the collected mushroom was identified as astraeus sp. based on the macroscopic and microscopic characteristics, biochemical reactions, and comparison with the standard keys of mushrooms (jordon, 1999; pacioni, 1981; phosri et al., 2007; purkayastha & chandra, 1985). molecular identification of fungal species the mushroom sample was identified as astraeus asiaticus using blast in the ncbi database based on query coverage (98.0%) and identity score (83. 05%) (figure 5) of sequence homology search. the nucleotide sequence of the its zone was submitted to genbank, ncbi (baltimore, usa) and published under the accession no mz931288.1. the strain name is “aa_skg.” phylogenetic analysis the phylogenetic tree (figure 6) revealed that the test organism was closely related to astraeus asiaticus. thus, the wild mushroom was concluded to be a member of astraeus asiaticus. 3.3. biodiversity analysis po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 10 https://en.wikipedia.org/wiki/hypha https://en.wikipedia.org/wiki/collenchyma the mushroom is terrestrial, grows under various trees, and is widely distributed in the three forests of bankura district, west bengal, india. astraeus asiaticus can establish a symbiotic relationship with a wide range of trees. this ectomycorrhizal species forms a symbiotic relationship with trees, such as shorea robusta, modhuca indica, petrocarpus marsupium, and terminalia bellrica. in our study, the abundance of basidioma in this species was found to be associated with the roots of shorea robusta; however, in the case of madhuca indica, a moderate number of this species was found. very poor or low numbers of this species were found in association with the roots of terminalia bellrica and petrocarpus marsupium. further, no mushroom basidioma was found in association with eucalyptus globulas and acacia auriculiformis trees (table 3). the ecological diversity of shorea robusta and madhuca indica in the 60 quadrats of 15 zones of three forests was calculated using the shannon diversity index formula. the diversity index of shorea robusta was 2.303 in the beliatore forest, 2.178 in the joypur forest, and 2.36 in the gangajal–ghati forest (table 4). further, the diversity index of madhuca indica was 2.25 in the beliatore forest, 2.26 in the joypur forest, and 2.29 in the gangajal–ghati forest (table 5). 3.4. mushroom extraction, mycochemistry, antioxidant content, and potentiality study mushroom extraction the hwe had the highest extraction yield (0.47 ± 0.01%) for a. asiaticus (table 6). the polarity index of water is 10.2. accordingly, water dissolves many polar and ionic solutes, such as proteins, water soluble carbohydrates, and mineral salts. in contrast, the nonpolar solvent, n-hexane, only dissolves nonpolar compounds, such as lipids, as its polarity index is 0.1, resulting in 0.32 ± 0.01b% crude product (he). acetone is a dipolar solvent; thus, it has an intermediate relative polarity. the polarity index of acetone is 5.1, resulting in a crude product, acetone extract (ae) of 0.42 ± 0.02%. the yields of hwe and ae were similar (p=0.005); however, their yields differed from that of he based on duncan multiple test analysis (table 6). mycochemical screening using a qualitative test based on the mycochemical screening results (table 7), the active ingredients in the hwe were flavonoids, cardiac glycosides, saponins, alkaloids, carbohydrates, carbohydrates, and phenols. for acetone, the active ingredients were flavonoids, cardiac glycosides, saponins, alkaloids, and phenols; and for the hexane extract, the active ingredients were terpenoids, cardiac glycosides, saponins, alkaloids, and tannins. phlobatannins and steroids were not detected in any sample. major active po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 11 ingredient metabolites, such as flavonoids, alkaloids, phenols, and carbohydrates, were found in the polar hwe, while terpenoids were found in the nonpolar hexane extract. antioxidant content total phenolics, flavonoid, and ascorbic acid content the total phenolics contents of the hot water, acetone, and hexane extracts were 6.8 ± 0.15, 3.95 ± 0.15, and 2.16 ± 0.26 mg gae/g, respectively, and the tfcs were 2.03 ± 0.12, 1.65 ± 0.2, and 1.01 ± 0.08 mg qe/g, respectively. figure 7 shows the ascorbic acid concentration in the mushroom extracts. the ascorbic acid content in the hot water, acetone, and hexane extracts was low (0.167 ± 0.03, 0.14 ± 0.04, and 0.11 ± 0.005 mg aae/g, respectively). antioxidant potential or scavenging activity based on the dpph method dpph was used to evaluate the rsa of the mushroom decoction. dpph is a stable free radical that exhibits idiosyncratic sorption at 517 nm. the absorption of these radicals decreases as antioxidants provide protons. as a measure of the magnitude of radical scavenging, a decrease in absorbance was confirmed. the free radical-scavenging capabilities of the extracts were calculated using the dpph assay. a method by which antioxidants prevent lipid oxidation is free radical scavenging. the distillate method for scavenging dpph free radicals can be used to assess the antioxidant activity of individual chemicals or extracts in a specific situation. figure 8 shows the dpph rsa of all solvent extracts of this mushroom. the ic50 (50% of inhibition) values for the hwe, acetone extract, and nonpolar hexane extract were 42.54 ± 1.25 µg/ml, 54.06 ±1.50 µg/ml, and 82.97 ± 1.58 µg/ml, while that of synthetic antioxidant bh was 32.41 ± 1.26 µg/ml. 4. discussion and conclusions wild edible mushrooms, which serve as a good economic source for underprivileged residents of bankura district, w.b., were collected to perform phenotypic and molecular identification studies and determine their diversity. this mushroom has been consumed as a good food in this area and adjoining districts. according to the literature, this genus of mushroom is distributed worldwide, especially in africa, asia, australia, europe, mexico, north america, and south america (pavithra et al., 2015). a literature survey revealed that astraeus hygrometricus, a. asiaticus, a. morganii, a. sirindhorniae, po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 12 a. odoratus, a. pteridis, a. smithii, a. telleriae, a. koreanus, and a. thailandicus have been discovered globally (verma et al., 2019). in this study, both phenotypic and molecular identification of this mushroom were successfully performed. the relevance of nucleic acids as molecular markers applies to fungal taxonomy. pcr is an established habitual tool for field mycology. for example, internal transcribed spacer (its) regions are amplified by pcr and subsequently sequenced for molecular phylogenetic analysis of diverse mushroom species. the dna sequences obtained via pcr are used for population genetics and biodiversity research on mushroom species (ghosh et al., 2022, izumitsu et al., 2012). in this study, both phenotypical and molecular (its1-5.8s-its-2 zone of rdna) identification revealed that this mushroom belonged to the species, astraeus asiaticus, which was first identified in this district of west bengal, india. astraeus asiaticus is a new species of star-shaped gasteroid fungus that differs morphologically from other genera of star-shaped fungi. this genus does not possess distinct peristome or columella. further, the spores of this genus are larger than those of geastrum, and its capillitial hyphae are longer and more branched (phosri et al., 2014). phosri et al. (2007) identified this species at the morphological and molecular levels. our phenotypical and molecular results align with the findings of phosri et al. (2007). recently, topno and srivastava (2021) collected puff balls from chota nagpur plateau (jharkhand). these puff balls were phenotypically and molecularly (itss markers) similar to astraeus asiaticus. similarly, vishal et al. (2021) collected two earth star mushrooms from sal forest of the porahat forest division of jharkhand state, india and identified these mushrooms a. asiaticus and a. odoratus based on molecular studies (itss marker). from the genbank database (ncbi, baltimore, usa), we identified five accession nos for astraeus sp. from 2008 to 2020: aj629895.1 for a. hygrometricus from west bengal, india in 2008; mn257431.1 and mt611066.1 for a. asiaticus from our neighboring state, jharkhand, india in 2019 and 2020; and mn262679.1 and kj847767.1 for astraeus odoratus in 2019 and 2016. sem studies of spores and hyphae were performed according to the findings of phosri et al. (2007). the study of biodiversity of this species of mushroom as ectomycorrhizan in this work in w.b. was first time work but in other states of india, diversity work has been done to some extent in other species (a. hygrometricus) (topno and srivastava, 2021; semwal et al., 2014; verma et al., 2017; vishal et al., 2021). after a distribution search of astraeus species in india, this genus was found in the provinces of himachal pradesh, uttar pradesh, uttarakhand, madhya pradesh, chhattisgarh, punjab, karnataka, kerala, jharkhand, and odisha, and in our state (pavithra et al., 2015; phosri et al., 2004; pyasi et al., 2013; semwal et al., 2014; topno & srivastava, 2021; vishal et al., 2021). some asian varieties of a. hygrometricus are reported to differ from those which are found in north america and are recognized as new species, such as a. asiaticus and a. odoratus (mortimer et al., 2012). in the present case, the astraeus species was identified as a. asiaticus. po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 13 in our study, the plants supporting this mushroom were shorea robusta, madhuca indica, petrocarpus marsupium, and terminalia belerica. such finding is supported by other researchers (harley et al., 1997; verma et al., 2017). the ectomycorrhizal association of astraeus with the roots of members of pinaceae, betulaceae, fagaceae, ericaceae, and dipterocarpaceae has been reported (wilson et al., 2012). the diversity index obtained in our study indicated that the ecological diversity of this mushroom was similar in the three forests in bankura district, w.b. by analyzing the soil quality of these three forests, they were found to be more or less similar in respect to npk and ph status, aligning with the results of another study (das, 2021). mushrooms contain high amounts of antioxidant compounds (chun et al., 2021; shaffiqueet et al., 2021; wani et al., 2010). the antioxidant activity of a. hygrometricus was demonstrated previously (mandal et al., 2015, pavithra et al., 2016). compounds, such as phenolic compounds, flavonoids, and ascorbic acids, have been found in other astraeus species (astraeus odoratus and astraeus pteridis) (arpha et al., 2012; isaka et al., 2016; lai et al., 2012; mallick et al., 2016; pimjuk et al., 2015; rahi & malik, 2016; stanikunaite et al., 2008). in this study, the different parameters of the mycochemistry study of a. asiaticus were successfully analyzed based on the total phenolic, flavonoid, ascorbic acid, and antioxidant contents, and the free rsa of this mushroom. in our experiment, all extracts of this mushroom had a high amount of phenolic compounds; however, the hwe had the best results. similarly, this hwe had the highest antioxidant activity among the extracts. the antioxidant properties depend on phenolic compounds (velioglu et al.,1998; xu, 2013). the dietary intake of a substantial amount of phenolic compounds is important as they play the role of free radical terminators that reduce the incidence of many diseases, such as atherosclerosis, cancer, and heart diseases (alothman et al. 2009; xu, 2013). the antioxidant activity of many wild mushrooms is positively correlated with total phenolics (lo & cheung, 2005; xu, 2013). phenolic compounds have redox properties and antioxidant properties. as their free radical scavenging ability is facilitated by their hydroxyl groups, the total phenolic concentration can be used as a basis for rapid screening of antioxidant activity. recently, bouyahya et al. (2022) reviewed the role of phenolic compounds in improving the chemosensitivity of anticancer drugs. this study first time has compared the antioxidant activity of polar, semipolar, and nonpolar extracts of astraeus asiaticus. so, mycochemical analyses should be performed to identify the active phenolic and flavonoid components. ascorbic acid plays an important role as a protective antioxidant. in this study, the ascorbic acid contents in the hot water, acetone, and hexane extracts were 0.167 ± 0.03, 0.14 ± 0.04, and 0.11 ± 0.005 aaes/g, respectively, aligning with previous results (mau et al, 2002). the highest ascorbic acid content of 0.167 ± 0.03 aaes/g, which was obtained using the hwe, might account for the better results found for their antioxidant activity. the hwe of a. asiaticus displayed potential scavenging activity (ic50 = 42.54 ± 1.25 µg/ml), which is comparable to that of the standard po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 14 antioxidant, bha. in general, the lower the ic50 value, the higher the antioxidant property. accordingly, the hwe of astraeus asiaticus was found to exhibit antioxidant attributes. these findings indicate that the hwe of mushroom species has a conspicuous effect on scavenging free radicals. pavitra et al. (2016) reported that the antioxidant activity of a. hygrometricus is concentration-dependent and does not decrease after cooking. herein, the hwe of a. asiaticus had the best antioxidant activity, which aligns with previous findings (pavitra et al., 2016). antioxidants are important compounds that help increase the resistance of the body to free radicals. mycochemistry revealed that this mushroom was the source of some volatile compounds, such as 1-octanol, 1-octen3-ol, and 1-octen-3-one, which had a "mushroom-like, earthy, and pungent odor that was evident as an oily and moss-like smell upon opening the caps." during the cooking of mushroom samples, furfural, benzaldehyde, cyclohexenone, and furanyl compounds are emitted (mortimer et al., 2012; verma et al., 2017). recently, isaka et al. (2017) isolated 17 known and seven new lanostane triterpenoids, such as astraeusins, 26-epi-artabotryol c1, and 26-epiastrasiaone, from a. asiaticus. to our knowledge, the current study is the first to assess the antioxidant content and activity of this edible mushroom species, and its biodiversity in the forests of bankura district. based on our findings, the species of astraeus in the forest of the bankura district was a. asiaticus. further, this species was mycorrhizally associated with shorea robusta and madhuca indica. based on the ecological diversity of this mushroom, a. asiaticus was associated with the above-mentioned specific plants. the mycochemistry of this mushroom revealed good quality and content of antioxidants, including phenolic compounds, flavonoids, and ascorbic acid, and good antioxidant activity (scavenging of free radicals). therefore, this mushroom is a good source of valuable nutrients, including antioxidants, scientifically validating our eating habits. this work may fill the knowledge gap, to some extent, related to this wild edible mushroom (gift of nature), and encourages further assessments. acknowledgements the authors would like to thank the department of science and biotechnology technology of west bengal, india for providing financial support. authors’ contributions skg designed the study and wrote and revised the manuscript. kp performed the experimental work, literature survey, and statistical analysis. po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 15 references alothman, m., bhat, r., & karim, a.a. 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(2013) anti-cancer effects of phenolic-rich extracts of button mushrooms (agaricus bisporus) phd thesis, department of food science, the graduate school, the pennsylvania state university, usa. 135. tables: table 1 mean values for the soil physico-chemical parameters and the n, p, and k ratios of the soil samples collected at selected quadrants of the forests in bankura district. name of the district name of the forests soil ph electrical conductivit y (µs) carbon (%) organic nitrogen kg/ha phosphorus kg/ha potassiu m kg/ha n:p:k bankura beliatore 6.5 86.72 0.93 220 74.34 215.7 3:1:3 gangajal –ghati(gghati) 6.3 84.29 0.87 140 73.54 214.2 2:1:3 joypur 6.2 81.58 0.82 180 72.67 218.4 2:1:3 po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 22 https://doi.org/10.20546/ijcmas.2019.801.063 https://doi.org/10.5897/jmpr09.565 https://doi.org/10.1111/j.1469-8137.2012.04109.x https://www.worldbank.org/en/topic/biodiversity, table 2 color change profile based on the fruiting bodies of astraeus asiaticus. table 3 association of astraeus asiaticus with different trees. note: indicates no association or mushroom fruit body, + indicates presence of a lower number of mushroom fruit bodies or association, ++ indicates a moderate number of fruit bodies, and +++ indicates an abundance of mushroom fruiting bodies. table 4 shannon weaver biodiversity index of the association between astraeus asiaticus and shorea robusta. test colour changed observed 10% koh none 4% hno3 none phenol none vanilin none 10% h2so4 none trees fruit body association with trees shorea robusta +++ petrocarpus marsupium + terminalia bellrica + acacia auriculiformis modhuca indica ++ eucalyptus globulas forest zones quadrates in each zone total no of quadrates in the forest shannon weaver biodiversity index of a. asiaticus beliatore 5 4 20 2.303 joypur 5 4 20 2.178 po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 23 table 5 shannon weaver biodiversity index of the association between astraeus asiaticus and modhuca indica. table 6 percentage yield of the crude products using different solvents. gangajal – ghati 5 4 20 2.36 forest zones quadrate in each zone total no of quadrates in the forest shannon weaver biodiversity index of a. asiaticus beliatore 5 4 20 2.25 joypur 5 4 20 2.26 gangajal – ghati 5 4 20 2.29 solvent name (extract name) method of extraction ratio of solute and solvent initial weight of mushroom powder(gm) final crude weight (gm)(±sd) percentage of yield (%) (±sd) hot water (hwe) dipping method 1:10 500 2.26± 0.15 0.47± 0.01a acetone (ae) dipping method 1:10 500 2.1± 0.12 0.42± 0.01a hexane (he) dipping method 1:10 500 1.52± 0.08 0.32± 0.02b po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 24 note: same letters in the same row indicate statistical similarity, while different letters indicate statistical difference (p=0.05) based on duncan’s multiple test. table 7 mycochemical screening of the polar (water), dipolar (acetone), and nonpolar (hexane) solvent extracts of astraeus asiaticus. note: (-) absence, (+) presence. mycocompound /class hot water extract (hwe) acetone extract (ae) hexane extract (he) flavonoid + + terpenoid + cardiac glycoside + + + steroid saponins + + + alkaloid + + + tanins + carbohydrate + phenol + + phlobatannins po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 25 figures figure 1 design of the mushroom fruitbody extraction process in different solvent systems po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 26 figure 1 images of astraeus asiaticus. a. young sporocarps collected from the field; b. young sporocarp; c. mature sporocarp. figure 3 images of the anatomy of astraeus asiaticus a. cross-section of the gleba; b. cross-section showing the arrangement of long thick-walled septate hyphal cells; c. spore mass; d. spore mass with basidiospore (40x). po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 27 figure 4 scanning electron micrograph. a. scanning electron micrograph of the hyphae structure (bar= 1 µm); b. scanning electron micrograph of mature basidiospores displaying dense, rounded spine in groups (bar = 200 nm). po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 28 figure 5 blast result of astraeus sp. with the highest sequence similarity of astraeus asiaticus. po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 29 figure 6 neighbor-joining phylogenetic tree showing the evolutionary relationship of the its gene sequences. the tree was constructed based on evolutionary distances, which revealed that the test organism is closely related to astraeus asiaticus sp. po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 30 figure 7 total phenol, flavonoid, and ascorbic acid contents of astraeus asiaticus in the hot water, acetone, and hexane extracts. po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 31 figure 8 ic50 value of the different solvent extracts of astraeus asiaticus (± sd is depicted at the top of data bar). similar letters at the top of data bars indicate statistical similarity, while different letters indicate statistical difference (p=0.05) based on duncan’s multiple range test. po st pr in t vol. 57, article 575 doi: 10.5586/am.575 acta mycologica: postprint version 32 a new report on hesperomyces coleomegillae (ascomycota, laboulbeniales) parasitism of coleomegilla maculata (coleoptera, coccinellidae) in brazil 1 of 7published by polish botanical society acta mycologica original research paper a new report on hesperomyces coleomegillae (ascomycota, laboulbeniales) parasitism of coleomegilla maculata (coleoptera, coccinellidae) in brazil carlos antonio inácio1*, hagabo honorato de paulo2, jonas dias almeida2, jessica rembinski2, elen lima aguiar menezes2, alessandra carvalho silva3 1 department of entomology and plant pathology, universidade federal rural do rio de janeiro, br 465, km 7, university campus, rural zone, seropédica, rj, cep 23851-970, brazil 2 department of entomology and plant pathology, rural university of rio de janeiro, ufrrj, br 465, km 7, university campus, rural zone, seropédica, rj, cep 23851-970, brazil 3 embrapa agrobiology, br 465, km 7, ecology site, rural zone, seropédica, rj, cep 23981-355, brazil * corresponding author. email: inacio@ufrrj.br abstract for the first time, the genus hesperomyces has been reported to infect coleomegilla maculata in laboratory mass rearing in brazil. thalli were found growing on several parts of this ladybird species, including the head, elytra, legs, and abdomen. infested adults died after 60 days. keywords host–parasitic interaction; laboulbeniomycetes; ectoparasitic fungi; 12-spotted ladybird; biotrophic parasites; tropical fungi introduction the order laboulbeniales (fungi: ascomycota) comprises about 2,200 species in 141 genera [1] and was previously included in basidiomycota and/or zygomycota. recent studies based on molecular data confirmed that laboulbeniales are members of ascomycota [2]. they are biotrophic ectoparasites of arthropods, and beetles (insecta: coleoptera) and flies (insecta: diptera) are their most common hosts [3–5]. these fungi are specialized and develop by forming a multicellular thallus in the tegument of living arthropods [6–8]. seven species of laboulbeniales parasitize ladybirds (coleoptera, coccinellidae) [6]: six species of the genus hesperomyces (h. chilomenes thaxt., h. coccinelloides thaxt., h. coleomegillae w. rossi & a. weir, h. papuanus t. majewski & k. sugiy., h. palustris w. rossi & a. weir, and h. virescens thaxt.) and one laboulbenia species (l. coccinellidicola haelew.) [6,9]. an eighth species, h. hyperaspidis thaxt., was brought into synonymy with h. virescens based on morphological characteristics [10], although this decision might have been premature [6]. coleomegilla maculata degeer (coleoptera: coccinellidae), the 12-spotted ladybird, is widely distributed in natural and managed ecosystems in north, central, and south american regions, including brazil [11–14]. it is primarily an aphidophagous insect, but its diet may include other insects (e.g., scales, psyllids, eggs, and neonate larvae of lepidoptera and coleoptera). thus, this ladybird is considered an important agent for crop pest control [15–17]. coleomegilla maculata adults are known to be hosts for h. coleomegillae and h. palustris in ecuador, costa rica, and cuba. the first observation of c. maculata acting as a host for hesperomyces (h. palustris) occurred in 1951 in cuba doi: 10.5586/am.1117 publication history received: 2018-02-25 accepted: 2018-11-21 published: 2019-04-18 handling editor malgorzata ruszkiewiczmichalska, institute for agricultural and forest environment, polish academy of sciences, poland authors’ contributions hh: collected and examined the material, drafted the manuscript; jd and jr: examined the material, prepared the figure; contributed to manuscript preparation; as: wrote the manuscript; em: wrote the manuscript and identified ladybird species; ca: examined and identified the material, wrote the manuscript funding the authors acknowledge capes, cnpq, and faperj for funds supporting this research. competing interests no competing interests have been declared. copyright notice © the author(s) 2019. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation inácio ca, de paulo hh, almeida jd, rembinski j, menezes ela, silva ac. a new report on hesperomyces coleomegillae (ascomycota, laboulbeniales) parasitism of coleomegilla maculata (coleoptera, coccinellidae) in brazil. acta mycol. 2019;54(1):1117. https:// doi.org/10.5586/am.1117 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:inacio%40ufrrj.br?subject=a%20new%20report%20on%20hesperomyces%20coleomegillae%20%28ascomycota%2c%20laboulbeniales%29%20parasitism%20of%20coleomegilla%20maculata%20%28coleoptera%2c%20coccinellidae%29%20in%20brazil https://doi.org/10.5586/am.1117 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1117 https://doi.org/10.5586/am.1117 2 of 7© the author(s) 2019 published by polish botanical society acta mycol 54(1):1117 inácio et al. / a report on hesperomyces parasitism of coleomegilla maculata [9]. coleomegilla maculata adults were found with h. palustris and h. coleomegillae thalli in costa rica and ecuador [18]. the brazilian laboulbeniales fungi have not yet been thoroughly investigated. there is a record of laboulbenia ecitonis blum from the eciton, or army ant, genus (hymenoptera: formicidae, ecitoninae) in curitiba, state of paraná [7,19]. data in previous literature indicates that nearly 100 species of laboulbenialean fungi have been recorded in brazil, the majority of which have been found on beetles [20,21]. in addition, rossi and bergonzo [21] documented the occurrence of 13 laboulbeniales species in brazil – all but one associated with beetles. the exception was h. coccinelloides, found on a ladybird, diomus seminulus (mulsant), in the state of ceará. this work aims to report hesperomyces infecting c. maculata adults in mass-rearing laboratory conditions in seropédica, rio de janeiro, brazil. material and methods a colony matrix of c. maculata was reared in a climate-controlled laboratory (25°c ±1°c, 60% ±10% rh, and 12-hour photoperiod) at the integrate pest management center (cimp) of the department of entomology and plant pathology (denf) of the universidade federal rural do rio de janeiro (ufrrj) on the seropédica campus in rj, brazil. this colony originated from adults collected from the organic farm fazendinha agroecológica km 07, also known as integrated system of agroecological production (sipa) in the municipality of seropédica, rj (22°45'24" s, 43°40'29" w) in 2010 [22]. adults and larvae were reared continuously and fed ad libitum on living larvae of drosophila melanogaster meigen (diptera: drosophilidae). the offspring of adults collected in this farm were introduced annually in the colony matrix to maintain the vigor of the colony. the field adults were separated from those of the colony matrix and held in plastic containers. adults were stored in disposable 1-liter transparent plastic containers sealed with organza to enable gas exchange. due to the difficulty in visually determining the sex [23], six adults were kept per container; however, when mating had not been observed in a 24-hour period, random exchanges among containers of some individuals were performed to ensure the presence of at least one viable pair per container. filtered water was provided using cotton wool placed in plastic bottle caps. the larvae of c. maculata were individually kept in 20-ml glass vials closed with hydrophilic cotton from the second instar until adulthood. from september to december 2015 and january to may 2016, c. maculata adults (6 months to 1-year old) with yellowish structures in some parts of their integument were observed in the colony matrix at cimp. each individual was observed under a dissecting microscope in the laboratory of mycology (denf, ufrrj). examining these adults revealed that the visible structures were thalli of laboulbeniales fungi. their position on the host integument was recorded (tab. 1). to identify the parasite, the thalli were gently removed from the host’s cuticle using a needle as well as dissecting and optical microscope techniques. the thalli were mounted on permanent slides stained with cotton-blue/ lactoglycerol or floxin/koh glycerol. measurements and pictures were taken using an olympus bx41 optical microscope with a digital camera and micrometer. the following morphological characteristics were determined: total length from foot to perithecial tip, length from foot to tip of uppermost antheridium, length and width of perithecium, and length and width of ascospores. both the philco-hitachi tm 1000 electron microscope, located at the health and sciences biological institute – icbs/ufrrj (seropédica campus), and evo ls 10 (carl zeiss), located at embrapa agrobiology (seropédica, rj), were tab. 1 number of mature thalli of h. colleomegillae on different body parts of several host individuals. body part number of thalli* % of thalli head 25 8.9 leg 44 16.0 thorax 46 30.1 abdomen 61 22.0 elytra 64 23.0 total 281 100.0 * taken from 50 naturally infected individuals. 3 of 7© the author(s) 2019 published by polish botanical society acta mycol 54(1):1117 inácio et al. / a report on hesperomyces parasitism of coleomegilla maculata utilized for some measurements, and pictures were acquired. identification of the fungal parasite followed de kesel [24] and goldmann et al. [18]. sampled material and voucher slides were deposited at the phytopathological herbarium verlande duarte silveira (denf/ufrrj), ufrj. results the fungus was identified in the genus hesperomyces, described by thaxter in 1891 (ascomycota, laboulbeniomycetes, laboulbeniales) [25]. hesperomyces coleomegillae w. rossi & a. weir (fig. 1) material examined. brazil, rio de janeiro, seropédica: main campus of ufrrj, inside a building of cimp, coordinates 22°46'10" s, 43°41'38" w, on elytra of the ladybird coleomegilla maculata degeer, november 14, 2015, h. h. paulo no. 3 (ufrrj – 12.305). the fungus formed ascomata on several parts of c. maculata adults, including the prothorax, mesothorax, and head (antennae and mouthparts, such as palpi); however, formation primarily occurred on the elytra (dorsal part) (fig. 1a–g). the fore, middle, and hind legs and abdomen also exhibited infection (tab. 1). ascomata showed the following characters: length from foot to top of perithecium: up to 770 μm; length from foot to tip of uppermost antheridium: 108–156 μm; perithecium: 92–563(–670) × 34–80(–96) μm, showing apical outgrowths (26–36 μm) (fig. 1j–l); ascospores: 68–96 × 5–6 μm, hyaline, one-septate, spindle-shaped, with thick a mucilaginous layer (fig. 1l) (tab. 2). discussion this paper reports the first record of living c. maculata adults hosting hesperomyces in laboratory mass-rearing in brazil, originating from specimens collected in the field. eggs and larvae were never infected, which was expected as laboulbeniales fungi only infect living adults [4,6]. this fungus was identified as h. coleomegillae based on morphological characters described in previous literature [18] (also see tab. 2). it has already been reported that species in the genus hesperomyces parasitize c. maculata adults; specifically, h. coleomegillae in central america (costa rica) and h. palustris in costa rica, cuba, and ecuador [18,26]. laboulbeniales fungi do not have a free-living stage, and the propagation of their sticky ascospores is triggered or promoted by the activity of the host (grooming, copulation, or other contact) [4,6,27]. a number of factors may have led the ladybird colony to become infected with h. coleomegillae. it is likely that a field-collected adult bearing mature thalli (having passed unnoticed by the collector) was enough to initiate propagation among the adults in the rearing containers. in addition, when mating had not been observed within 24 hours, there was random exchanges of individuals among the containers until the presence of at least one viable pair per container was observed. however, new adult collections should be carried out on the same farm to more accurately detect natural infection of c. maculata by hesperomyces species. individual cycloneda sanguinea (linnaeus), eriopis connexa (germar), harmonia axyridis (pallas), and hippodamia convergens guerin-meneville adults have also been reported to occur at sipa [13,28,29], and h. virescens thaxt. has been reported to infect adult lady beetles of the above species in other countries [6,25]. therefore, new records of species in hesperomyces on wild populations of these lady beetles in brazil should be expected. however, based on studies by cottrell and riddick [30] and riddick [31] the chances of transmission taking place from h. virescens-infected h. axyridis to c. maculata adults at sipa are likely low. these authors demonstrated that intraspecies transmission of this fungus in h. axyridis was common, whereas interspecies transmission of h. axyridis to coccinella septempunctata l. and olla v-nigrum (mulsant), as well 4 of 7© the author(s) 2019 published by polish botanical society acta mycol 54(1):1117 inácio et al. / a report on hesperomyces parasitism of coleomegilla maculata fig. 1 (a–g) body parts of coleomegilla maculata infected by hesperomyces. (a) pronotum and dorsal part of the elytra of c. maculata with thalli (bar: 1 mm). (b,c) legs and prosternum of c. maculata with ascomata (bar: 500 µm). (d–f) pictures obtained from a scanning electron microscope (sem). (d) detail of the fungus on the head and pronotum of c. maculata (bar: 5 mm). (e) infection on leg (bar: 600 µm). (f) detail of infection of tibia (bar: 600 µm). (g) detail of infection, mainly of femur (bar: 600 µm). (h–l) hesperomyces structures. (h) detail of young antheridium with appendages (bar: 15 µm). (i) apical outgrowth of perithecia (bar: 100 µm). (j,k) perithecia (bar: 100 µm). (l) ascospores (bar: 20 µm). 5 of 7© the author(s) 2019 published by polish botanical society acta mycol 54(1):1117 inácio et al. / a report on hesperomyces parasitism of coleomegilla maculata as from o. v-nigrum to both c. septempunctata l. and h. convergens guerin-meneville, was notably uncommon. the same authors [30,31] did not observe infection of c. maculata adults after confinement with h. axyridis after placed in a vial and tumbled on a vial roller for 1 h. in general, laboulbeniales fungi cause little or no harm to their arthropod hosts and do not seem to kill them [32–35]. however, there are indications that species in the genus hesperomyces deviate from this because of the penetrating haustorium in the exoskeleton of their hosts, and some negative effects on hosts have been reported [36,37]. hosts with high numbers of thalli, such as those from h. virescens on h. axyridis with more than 100 thalli (sometimes >400 thalli), on the outer parts of their body (e.g., elytra, eyes, antennae, mouthparts, and/or legs) may no longer be able to fly, mate, or detect their prey [6,37]. in this study, it was observed that 100% of naturally colonized adults died after 60 days and the number of eggs laid by infected adults was affected compared to noninfected adults of the same age. it was also observed that c. maculata adults under 6 months of age were not infected by h. coleomegillae. this result suggests that older adults may be more susceptible to infection by this species. additional research is needed to support this hypothesis. tab. 2 comparison of morphological characters (in µm) of hesperomyces species associated with coccinellidae (coleoptera) reported from central and south america. species country length from foot to perithecial tip length from foot to tip of uppermost antheridium length and width of perithecium perithecial outgrowth length and width of ascospores h. coccinelloides1 panama 120–240 24–30 × 85–110 35–40 h. coleomegillae2 costa rica, ecuador 370–600 130–150 55–70 × 210–350 27–33 75–77 h. hyperaspidis3 trinidad tobago 180 25 × 110 20 h. palustris2 costa rica, ecuador (355)400–675 105–130 55–70 × 245–355 50–70 h. coleomegillae4 brazil up to 770 108–156 34–80(–96) × 92–563(–670) 26–36 68–96 × 5–6 1 thaxter [38] (host: scymnus tardus mulsant); 2 goldmann et al. 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wrocław university of environmental and life sciences, poland; https://orcid.org/0000-00017236-8005 funding this work did not involve any funding. competing interests no competing interests have been declared. copyright notice © the author(s) 2020. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. editorial introduction wojciech pusz * division of plant pathology and mycology, department of plant protection, wrocław university of environmental and life sciences, pl. grunwaldzki 24a, wrocław, 50-363, poland *to whom correspondence should be addressed. email: wojciech.pusz@upwr.edu.pl dear mycologists, it is an honor for us to be presenting you with this special issue of acta mycologica, which is dedicated to two great scientists, prof. dr hab. maria dynowska and prof. dr hab. maria rudawska. both of them have made significant contributions to the world of mycological research, and their impacts continue in the daily works of many research centers that base their foundations on prof. dynowska’s legacy and prof. rudawska’s constructs. looking at prof. maria dynowska’s career path, there is no doubt that, as a scientist, she particularly valued interdisciplinary studies, where she focused on the ecology of fungi, with special attention to their variable nature and the consequences thereof. prof. dynowska has conducted numerous studies on aquatic ecosystems, tracing the natural reservoirs of anthropopathogens and searching for microorganisms that can potentially be used as bioindicators for assessing both the sanitary quality of water and epidemic threats. through her studies on crop phytopathogens, such as typhula spp., fusarium spp., and common saprotrophs, her research has enriched our knowledge on phytopathology immensely. finally, she devoted several explorations to investigating human mycobiota, specifically the roles and potentials of human pathogens present on skin and in the digestive tract, thereby enhancing our understanding of medical mycology. her legacy reaches beyond the university’s walls, leaving a solid mark on medicine, education, and ecological knowledge. with a focus on ectomycorrhizal fungi associated with forest trees, prof. maria rudawska’s many years of research work have contributed to the identification of more than 80 fungal species in this environment. she has carried out numerous studies on the diversity of ectomycorrhizal fungi in bare-root forest nurseries. her work showing that such forests are well colonized by these fungi has had direct impacts on both the management of these forests and improvement of the health conditions of woodlands. this illustrates how prof. rudawska’s scientific interests have evolved. after a long period of intensive research on the physiology of the ectomycorrhiza, she switched her attention to the problem of forest decline and the roles of mycorrhizae and mycorrhizal fungi in this process. her legacy is still in the making as she continues to inspire her coworkers and young professionals to devote their time to uncovering the secrets of the ectomycorrhizal relationship. i am aware that the articles in this issue will not fully reflect the enormity of the work, the insights in analyzing the results obtained, and the commitments that characterize both professors. however, i hope that they will become an inspiration for young adepts of mycology and serve as a supplement of knowledge for those who already have experience in scientific work. acta mycologica / 2020 / volume 55 / issue 2 / article 55213 publisher: polish botanical society 1 https://doi.org/10.5586/am.55213 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0003-1531-2739 mailto:wojciech.pusz@upwr.edu.pl verrucaria species and other rare amphibious lichens in the beskid sądecki mts 1 of 8published by polish botanical society acta mycologica original research paper verrucaria species and other rare amphibious lichens in the beskid sądecki mts natalia matura*, beata krzewicka laboratory of lichenology, w. szafer institute of botany, polish academy of sciences, lubicz 46, 31-512 kraków, poland * corresponding author. email: n.kapek@botany.pl abstract ten freshwater lichen species from the beskid sądecki mts are presented. seven of them: hydropunctaria rheitrophila, thelidium aquaticum, t. minutulum, t. zwackhii, verrucaria dolosa, v. elaeomelaena and v. submersella, are new to the region. three species: verrucaria elaeina, v. hydrophila and v. latebrosa, were previously known from single localities. keywords freshwater lichens; lichenized fungi; verrucariaceae; carpathian mts this issue of acta mycologica is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. introduction lichenized fungi occurring in freshwater ecosystems comprise a small (about 5% of the world’s population) and poorly known group of organisms compared to the terrestrial lichens. they are restricted to submerged or partially inundated rocks, such as in springs, rivers, and lakes. because of their biology, aquatic and semi-aquatic lichens constitute a very interesting ecological group of fungi. in freshwater habitats, species distribution is known to be affected by several ecological factors related to the length of submergence, shading, substrate (lithology, stability), water chemistry, speed and transportation [1–3]. freshwater lichens belong to a few genera, the most representative being verrucaria. species of the family verrucariaceae occur in nearly all european freshwater lichen communities, being often the dominant or the only family occurring in the permanently submerged zone [4]. lichenological research in the beskid sądecki mts was initiated by rehman [5] and boberski [6] at the end of the 19th century. the first records regarding aquatic species were reported by olech [7,8], who conducted a comprehensive survey of the lichen biota of the area and recognized five aquatic taxa: bacidina inundata (fr.) vězda, verrucaria aquatilis mudd, v. denudata zschacke (at present v. hydrophila orange), v. guestphalica servít (at present v. elaeina borrer) and v. laevata körb. [at present v. praetermissa (trevisan) anzi]. a comparative survey was conducted by śliwa [9] in the beskid sądecki mts in the late 1990s in order to determine the impact of human doi: 10.5586/am.1057 publication history received: 2015-04-15 accepted: 2015-06-23 published: 2015-08-05 handling editor maria rudawska, institute of dendrology of the polish academy of sciences, poland authors’ contributions nm: field research and draft of the manuscript; bk: species identification and critical revising; bk, nm: final writing of the manuscript funding the study was supported by the national science centre, grant no. n n304 170539 and from the w. szafer institute of botany, polish academy of sciences through its statutory funds. competing interests no competing interests have been declared. copyright notice © the author(s) 2015. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation matura n, krzewicka b. verrucaria species and other rare amphibious lichens in the beskid sądecki mts. acta mycol. 2015;50(1):1057. http://dx.doi. org/10.5586/am.1057 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:n.kapek%40botany.pl?subject=verrucaria%20species%20and%20other%20rare%20amphibious%20lichens%20in%20the%20beskid%20s%c4%85decki%20mts http://dx.doi.org/10.5586/am.1057 http://creativecommons.org/licenses/by/3.0/ http://creativecommons.org/licenses/by/3.0/ http://dx.doi.org/10.5586/am.1057 http://dx.doi.org/10.5586/am.1057 2 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1057 matura and krzewicka / amphibious lichens in the beskid sądecki mts activity on the diversity and distribution of lichens. śliwa [9] confirmed the occurrence of two freshwater taxa (b. inundata and v. praetermissa) reported previously. additionally, she reported one more species, that is verrucaria anziana garov. (at present v. latebrosa körb.). furthermore, a few new localities of lichens associated with freshwater habitats were reported from the żebracze nature reserve by czarnota [10]. however, permanently and periodically submerged habitats were not of special concern in the studies mentioned above. in 2013, a lichenological survey focused on the lichen biota of freshwater habitats was carried out by us in the beskid sądecki mts. a considerable collection of lichens associated with such habitats was obtained in the survey. many interesting species, including new regional records, were identified in the material collected. material and methods field work was carried out by the first author in the beskid sądecki mts in 2013. the following streams were included in the study: baraniecki stream, czaczowiec stream, szczawniczek stream, uhryński potok stream, wierchomlanka stream and wojkowski stream. the lichen material was analyzed with standard morphological and anatomical methods using microscopic techniques. voucher specimens are available in the herbarium of the w. szafer institute of botany, polish academy of sciences (kram). results and discussion ten freshwater lichen species were recorded by us in the beskid sądecki mts. seven of them are new to the region (hydropunctaria rheitrophila, thelidium aquaticum, t. minutulum, t. zwackhii, verrucaria dolosa, v. elaeomelaena, v. submersella). three species (verrucaria elaeina, v. hydrophila and v. latebrosa) were known in the study area only from single localities. hydropunctaria rheitrophila (zschacke) keller, gueidan & thüs verrucaria rheitrophila zschacke it is characterized by completely immersed perithecia visible as black dots on the upper surface of the thallus and by a greenish thallus with black punctae giving a spotted and weakly roughened appearance on the upper surface. black punctae are well visible in thinner thalli; however, they are sometimes completely immersed in the thicker ones and then the upper surface is even. ascospores are simple, colorless and ellipsoid reaching up to 10–12(–15) × 4.5–7 µm. hydropunctaria scabra is another freshwater species with black punctae in the thallus but it differs in the larger perithecia forming projecting mounds, larger ascospores 14–17 × 7.5–9 μm, and the presence of a greenish black to black thallus with a continuous black basal layer. hydropunctaria rheitrophila occurs on submerged siliceous or calcareous rocks and pebbles in sunny places. this species is reported from the beskid sądecki mts for the first time. it was found in the szczawniczek stream on submerged stones in a sunny place at an altitude of 900 m. it is widespread in the mountainous regions of southern poland, where it has many localities, especially in the western carpathians, where it occurs in nearly all mountain ranges [11]. this species was also recorded from lower altitudes at scattered localities in central and northern poland [11,12]. in europe, it is widespread from sea level to alpine areas [9]. specimens examined. poland. western carpathian mts, beskid sądecki mts, pasmo jaworzyny krynickiej range: szczawniczek stream, on left side of road, before turning 3 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1057 matura and krzewicka / amphibious lichens in the beskid sądecki mts onto the trail towards the runek peak, 49°25'385" n, 20°52'476" e, alt. 945 m, on submerged stone in sunny place, 5 september 2013, leg. n. matura (kram). thelidium aquaticum servít it is characterized by a superficial, thin, dark olive greenish to brownish thallus which is continuous or with small cracks. perithecia are initially completely immersed in the thallus, their upper half becoming exposed later. the involucrellum is absent. the exciple is brown-black above and colorless to pale brown at base, reaching up to 80–115 μm in diameter. ascospores are 3-septate, reaching up to 16–27 × 5–8 μm. thelidium zwackhii also has 3-septate ascospores but differs in the bigger exciple reaching up to 140–300 μm wide. thelidium aquaticum is an amphibious species occurring on siliceous substrate in streams. this species is reported from the beskid sądecki mts for the first time. it was found in the czaczowiec stream on inundated rocks in sunny places, at an altitude of 430 m. in poland, it is known from scattered localities in the carpathian mts [13] and from a few sites in central poland [14]. this species is poorly known in europe and outside poland it has so far been reported only from the type locality [4]. specimens examined. poland. western carpathian mts, beskid sądecki mts, pasmo jaworzyny krynickiej range: czaczowiec stream, near bus stop in izgierkówka village, 49°31'960" n, 20°48'464" e, alt. 434 m, on rock often inundated with water in sunny place, 27 july 2013, leg. n. matura (kram). thelidium minutulum körb. thelidium acrotellum arnold this species has a thin, superficial, pale grey-green to dark brown thallus. it is continuous, usually not cracked or forms numerous small patches. perithecia are prominent, without an involucrellum. they are very prominent or up to half-immersed in the thallus, with an inconspicuous ostiole. the exciple is brown above and usually colorless at base, reaching up to 80–240 μm in diameter. ascospores are 1-septate, reaching up to 13–32 × 4–15 μm. thelidium zahlbruckneri servít also has 1-septate ascospores but differs in the dark brown exciple reaching up to 100–150 μm in diameter and smaller ascospores 10–15 × 4–7 μm. thelidium zwackhii and t. aquaticum differs in the 3-septate ascospores. thelidium minutulum occurs mainly in non-aquatic (terrestrial) habitats on both calcareous and non-calcareous substrates or rarely on soil, in moist and shady places but it is also often found on small stones beside streams in the splash zone. this species was not previously reported from the beskid sądecki mts. it was found at scattered localities in the czaczowiec stream, the uhryński potok stream and the wojkowski stream. the species occurred on splashed or inundated substrates, in both sun-exposed and shaded places. in poland, it is known mainly from terrestrial populations from scattered localities in mountains and lowlands [13,15]. amphibious populations are often overlooked. the species is widespread in europe [4]. specimens examined. poland. western carpathian mts, beskid sądecki mts, pasmo jaworzyny krynickiej range: czaczowiec stream, near bus stop in czaczów village, 49°30'499" n, 20°47'119" e, alt. 562 m, on rock often inundated with water, 27 july 2013, leg. n. matura (kram); czaczowiec stream, near bus stop in izgierkówka village, 49°31'960" n, 20°48'464" e, alt. 434 m, on rock often inundated with water in sunny place, 27 july 2013, leg. n. matura (kram); uhryński potok stream, near shrine in uhryń village, 49°30'217" n, 20°51'647" e, alt. 539 m, on splashed rock in shaded place, 24 august 2013, leg. n. matura (kram); góry leluchowskie mts: wojkowski stream, a few hundred meters beyond the asphalt road, near clearing in wojkowa village, 49°20'227" n, 20°59'368" e, alt. 689 m, on splashed rock, 4 september 2013, leg. n. matura (kram). 4 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1057 matura and krzewicka / amphibious lichens in the beskid sądecki mts thelidium zwackhii (hepp) a. massal. this species is characterized by a thin, superficial, grey-green to dark brown, scattered thallus, in small flecks or forming a continuous or slightly cracked crust. perithecia are prominent, without an involucrellum, sessile or half-immersed in the thallus. the exciple is 140–300 μm wide, brown above and usually colorless at base. ascospores are 3-septate, reaching up to 25–32 × 10–12 μm. thelidium aquaticum also has 3-septate ascospores but differs in the smaller exciple reaching up to 80–150 μm in diameter and smaller ascospores 16–27 × 5–8 μm. thelidium zwackhii occurs mainly in non-aquatic (terrestrial) habitats but its amphibious populations are often noted by streams. this species is reported from the beskid sądecki mts for the first time. it was found in the uhryński potok stream on inundated rocks in sunny places, at 500 m. in poland, it is rather rare species [15,16]. in europe it is rarely recorded but presumably widely distributed [4]. specimens examined. poland. western carpathian mts, beskid sądecki mts. pasmo jaworzyny krynickiej range: uhryński potok stream, on the geological trail, 49°30'809" n, 20°51'898" e, on rock often inundated with water in sunny place, alt. 500 m, 24 august 2013, leg. n. matura (kram). verrucaria dolosa hepp this species is characterized by an almost absent or thinly superficial thallus which is non-gelatinous, 25–50 µm thick, green to olive-brown, more or less smooth, glossy, continuous and never areolate, only sometimes with a few cracks. perithecia form low to moderate projections 100–150(–180) µm wide; they are semi-immersed to prominent. they often are covered by the thallus in the lower part. the involucrellum is present, more or less conical. ascospores are simple, colorless, 15–17.5 × 6.5–8.5 µm. verrucaria maculiformis is similar in the presence of a thin thallus but differs in the larger perithecia forming moderate projections 150–250 µm in diam. and terrestrial habitat. verrucaria dolosa occurs on siliceous rocks, limestone and concrete, in moist habitats by freshwater watercourses, e.g., in the splash zone. this species is reported from the beskid sądecki mts for the first time. it was found in the baraniecki and szczawniczek streams on rocks in the splash zone, at altitudes of ca. 500 and 1000 m. the species occurs at scattered localities in the polish carpathian mts and in northern and central poland [11]. this species is widespread in europe [11]. specimens examined. poland. western carpathian mts, beskid sądecki mts, pasmo jaworzyny krynickiej range: baraniecki stream, 49°24'914" n, 20°48'058" e, alt. 508 m, in splash zone on rock, 7 july 2013, leg. n. matura (kram); szczawniczek stream, on left side of road, before turning onto the trail towards runek peak, 49°25'385" n, 20°52'476" e, alt. 945 m, on submerged stone in sunny place, 5 september 2013, leg. n. matura (kram). verrucaria elaeina borrer verrucaria guestphalica auct. this species is characterized by a whitish or grey-green rimose thallus with perithecia one-quarter to three-quarters immersed, rarely completely immersed in the thallus. perithecia usually vary in the same specimen, forming moderate projections 220–400 μm in diam. the involucrellum is well-developed, darkly pigmented, weaker colored in basal parts, conical-hemispherical to conical, usually more or less spreading from the exciple below. ascospores are simple, colorless, ellipsoid to narrowly ellipsoid or oblong-ellipsoid, 18–22(–24) × 7–9 μm. 5 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1057 matura and krzewicka / amphibious lichens in the beskid sądecki mts verrucaria praetermissa differs in the more immersed perithecia, slightly larger ascospores and the presence of a black basal layer. verrucaria submersella differs in the larger ascospores (up to 20–32 μm long), and v. sublobulata has smaller perithecia forming low projections 80–120 µm in diam., and a greenish thallus. verrucaria elaeina is a facultatively amphibious species occurring on a variety of substrates such as limestone, sandstone, and concrete. it occurs in shady places both in moist and dryer habitats but never at sunny and xerothermic sites. in the beskid sądecki mts, this species was previously reported by olech [8] at one locality in a stream in the jaworzyna krynicka range. it was found in our study at scattered localities in all the streams examined, mainly in the splash zone, at altitudes ranging from 500 to 600 m. the species is very common in the polish carpathians, where it was recorded in most of the ranges [11]. known in europe from scattered localities [11]. specimens examined. poland. western carpathian mts, beskid sądecki mts, pasmo jaworzyny krynickiej range: baraniecki stream, 49°25'283" n, 20°47'523" e, alt. 514 m, on rock often inundated with water in sunny place, 12 july 2013, leg. n. matura (kram); czaczowiec stream, in barnowiec village, beyond barrier and behind sharp bend in asphalt road, 49°29'592" n, 20°46'730" e, alt. 647 m, on rock often inundated with water in sunny place, 27 july 2013, leg. n. matura (kram); czaczowiec stream, near bus stop in czaczów village, 49°30'499" n, 20°47'119" e, alt. 562 m, on splashed rock, 27 july 2013, leg. n. matura (kram); uhryński potok stream, in uhryń village, at the level of the nature trail, 49°29'740" n, 20°51'641" e, alt. 576 m, on splashed rock in sunny place, 24 august 2013, leg. n. matura (kram); wierchomlanka stream, in wierchomla mała village, about 500 m behind last house on the road, 49°25'398" n, 20°49'269" e, alt. 612 m, on big stone often inundated in sunny place, 2 september 2013, leg. n. matura (kram); góry leluchowskie mts: wojkowski stream, a few hundred meters beyond asphalt road, near clearing in wojkowa village, 49°20'227" n, 20°59'368" e, alt. 689 m, on rock often inundated with water, 4 september 2013, leg. n. matura (kram); wojkowski stream, in wojkowa village, near historic orthodox church and bus stop, 49°20'758" n, 20°59'778" e, alt. 620 m, on splashed rock, 4 september 2013, leg. n. matura (kram). verrucaria elaeomelaena (a. massal.) arnold this species has a light brownish green to mid-brown subgelatinous thallus with perithecia at first completely covered by the thallus layer but becoming erumpent in thalline warts later. the apex is often somewhat exposed and blackish. the involucrellum is black, variable, conical, present in the upper half of the exciple or reaching to the base of the thallus. ascospores are simple, colorless, broadly ellipsoid to ovoid, rounded at both ends, 22–30 × 12–16 μm. it is distinguished from v. funckii, a species similar in appearance, by a thicker and paler thallus and larger, broadly ellipsoid ascospores (v. funckii has ascospores ellipsoid to narrowly ellipsoid, rounded at apices 18–25 × 6–10 μm, and occurs exclusively on siliceous rocks). verrucaria elaeomelaena is an amphibious species occurring on calcareous rocks, or rarely on sandstone or rarely on siliceous rocks submerged in calcareous water (with ph > 7), on inundated or submerged rocks and pebbles in streams mainly in lowlands and uplands, less frequently in mountains. this species is reported from the beskid sądecki mts for the first time. it was found in the czaczowiec and wojkowski streams on submerged stones in sunny places at an altitude of ca. 600 m. the species occurs at scattered localities in the polish carpathian mts and in northern and central poland [11]. in europe it is widespread but rare [4]. specimens examined. poland. western carpathian mts, beskid sądecki mts, pasmo jaworzyny krynickiej range: czaczowiec stream, in barnowiec village, beyond barrier and behind sharp bend in asphalt road, 49°29'592" n, 20°46'730" e, alt. 647 m, on rock often inundated with water in sunny place, 27 july 2013, leg. n. matura (kram); 6 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1057 matura and krzewicka / amphibious lichens in the beskid sądecki mts góry leluchowskie: wojkowski stream, in the central part of wojkowa village, sunny place near the road, 49°21'802" n, 20°58'935" e, alt. 555 m, on submerged stone, 4 september 2013, leg. n. matura (kram). verrucaria hydrophila orange verrucaria denudata zschacke nom. illeg., non nyl. (1858) verrucaria hydrela auct., non ach. (1814) the species is distinguished by a smooth uncracked, continuous, subgelatinous thallus without a black basal layer, perithecia covered by a layer of thallus, and a conical involucrellum reaching the base – on dry thalli visible as black points, on wet thalli visible as a black disc within the transparent thallus. ascospores are simple, colorless, ellipsoid, reaching up to 20–25(–26) × 10–12(–15) µm. initially this taxon was mistakenly reported as verrucaria hydrela ach. in poland. however, v. hydrela described by acharius [17] differs in the presence of an uneven, non-gelatinous thallus with a few cracks. for this reason, the specimens with a smooth, uncracked and subgelatinous thallus and with perithecia with a conical involucrellum covered by a layer of thallus were recognized by krzewicka [11] as v. denudata zschacke. unfortunately, this name was illegitimate therefore orange [18] described this taxon as v. hydrophila. verrucaria hydrophila is a freshwater species occurring on permanently submerged siliceous rocks, in sunny places. in the beskid sądecki mts, this species was previously reported at two localities from the pasmo radziejowej range [7] and at one locality from the pasmo jaworzyny krynickiej range [10]. in our study, this species was found on scattered localities in the baraniecki, czaczowiec and uhryński potok streams on submerged or often inundated stones, at altitudes ranging from 500 to 650 m. this species has many localities in mountainous regions of poland. it is widespread in the carpathian mts but also occurs in the sudeten mts, central and northern poland [11]. in europe, it is widespread but known as v. hydrela [11]. specimens examined. poland. western carpathian mts, beskid sądecki mts, pasmo jaworzyny krynickiej range: baraniecki stream, 49°24'914" n, 20°48'058" e, alt. 508 m, in the splash zone on rock, 7 july 2013, leg. n. matura (kram); czaczowiec stream, in barnowiec village, about 100 m beyond barrier, 49°29'372" n, 20°46'672" e, alt. 785 m, on rock often inundated with water in sunny place, 24 july 2013, leg. n. matura (kram); czaczowiec stream, near bus stop in izgierkówka village, 49°31'960" n, 20°48'464" e, alt. 434 m, on rock often inundated with water in sunny place, 27 july 2013, leg. n. matura (kram); uhryński potok stream, in central part of uhryń village, sunny place near asphalt road and human settlements, 49°28'604" n, 20°51'580" e, alt. 641 m, on submerged stone in sunny place, 23 august 2013, leg. n. matura (kram); uhryński potok stream, in uhryń village, at the level of the nature trail, 49°29'740" n, 20°51'641" e, alt. 576 m, on submerged stone in sunny place, 24 august 2013, leg. n. matura (kram). verrucaria latebrosa körb. this species is distinguished by a non-subgelatinous, well-developed thallus with many cracks or regularly areolate and almost completely immersed perithecia. perithecia form low to moderate projections, 250–500 μm in diam., mostly covered by the thallus except for the uppermost part with a black exposed convex ape. the involucrellum is well-developed (thick), present only around the apex or spreading outwards and downwards in the upper part. ascospores are simple, colorless, ellipsoid, (18–)24–29(–36) × 8.5–12.5(–14) μm, with a halo in the fresh material. verrucaria submersella differs in the more prominent perithecia, half or threequarters immersed in the thallus and forming shallow pits in the substrate, and the non-halonate ascospores. verrucaria cernaensis differs in the well-developed involucrellum reaching to the base of the dark pigmented exciple, and smaller ascospores 7 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1057 matura and krzewicka / amphibious lichens in the beskid sądecki mts [18–22(–25) × 8–14 μm]. verrucaria margacea differs in the prominent perithecia forming moderate to distinct projections (280–)350–800 μm in diam., at first covered by the thallus and later partly exposed, and a conical involucrellum reaching to the base of the exciple. verrucaria latebrosa is an amphibious species occurring on siliceous rocks beside streams and lakes, often above the water level but in the splash zone, in sunny or partially shady places. in the beskid sądecki this species was previously reported at one locality in a stream in the pasmo radziejowej range [9]. in our study, it was found at two localities in the czaczowiec and uhryński potok streams in the splash zone. in poland, it occurs in the southern part in the sudeten mts and the carpathian mts, mainly in the tatra mts [11]. in europe, it has been reported only from central europe to date [19]. specimens examined. poland. western carpathian mts, beskid sądecki mts, pasmo jaworzyny krynickiej range: czaczowiec stream, near bus stop in czaczów village, 49°30'499" n, 20°47'119" e, alt. 562 m, in splash zone on rock, 27 july 2013, leg. n. matura (kram); uhryński potok stream, near shrine in uhryń village, 49°30'217" n, 20°51'647" e, alt. 539 m, on rock often inundated with water in shaded place, 24 august 2013, leg. n. matura (kram). verrucaria submersella servít this species is characterized by a thin superficial to semi-endolithic dirty white to yellowish green cracked thallus without a black basal layer. perithecia are half or three-quarters immersed in the thallus, forming moderate projections raised above the thallus, naked in the upper part, forming shallow pits in the substrate. the involucrellum is present in the upper half of the exciple, rarely reaching to the thallus base, appressed to the exciple or slightly laterally spreading into the thallus. the exciple is colorless to pale brown. ascospores are ellipsoid, (20–)24–32 × 9–14 µm, without a halo. verrucaria praetermissa differs in the presence of a black basal layer. verrucaria sublobulata is similar by the color of the thallus but differs in smaller and nearly immersed perithecia. verrucaria submersella occurs in the splash zone on limestone and dolomite mainly in shady places. it is also recorded on siliceous rocks and pebbles in streams with hard, well-buffered water [4]. this species is reported from the beskid sądecki mts for the first time. it was found at one locality in the wojkowski stream on rocks often inundated with water. it is poorly known in poland, confirmed in the carpathian mts and in the central part of poland in the wyżyna krakowsko-wieluńska upland at scattered localities [11]. the species is reported from scattered localities in central europe from montane to subalpine areas [4]. specimens examined. poland. western carpathian mts, beskid sądecki mts, góry leluchowskie: wojkowski stream, a few hundred meters beyond asphalt road, near clearing in wojkowa village, 49°20'227" n, 20°59'368" e, alt. 689 m, on rock often inundated with water, 4 september 2013, leg. n. matura (kram). references 1. krzewicka b, galas j. ecological notes on verrucaria aquatilis and v. hydrela in the polish tatry mountains. in: lackovičová a, guttová a, lisická e, lizoň p, editors. central european lichens – diversity and threat. ithaca, ny: mycotaxon ltd.; 2006. p. 193–204. 2. nascimbene j, nimis pl. freshwater lichens of the italian alps: a review. ann limnol. 2006;42(1):27–32. http://dx.doi.org/10.1051/limn/2006003 http://dx.doi.org/10.1051/limn/2006003 8 of 8© the author(s) 2015 published by polish botanical society acta mycol 50(1):1057 matura and krzewicka / amphibious lichens in the beskid sądecki mts 3. nascimbene j, thüs h, marini l, nimis l. early colonization of stone by freshwater lichens of restored habitats: a case study in northern italy. sci total environ. 2009;407(18):5001– 5006. http://dx.doi.org/10.1016/j.scitotenv.2009.06.012 4. thüs h, schultz m. freshwater flora of central europe. fungi: 1st part: lichens. heidelberg: spektrum; 2009. 5. rehman a. systematyczny przegląd porostów znalezionych dotąd w galicji zachodniej opracowany na podstawie własnych i cudzych spostrzeżeń. spraw kom fizjogr pau. 1879;13:3–66. 6. boberski w. systematische übersicht der flechten galiziens. verh zool bot ges wien. 1886;36:243–286. 7. olech m. porosty pasma radziejowej. fragm flor geobot. 1972;18(3–4):359–398. 8. olech m. porosty beskidu sądeckiego. zeszyty naukowe uj prace botaniczne. 1973;1:87–192. 9. śliwa l. antropogeniczne przemiany lichenoflory beskidu sądeckiego. kraków: instytut botaniki uj; 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(prace botaniczne; vol 31). 10. czarnota p. porosty rezerwatu żebracze w beskidzie sądeckim. parki narodowe i rezerwaty przyrody. 2002;21(4):385–410. 11. krzewicka b. a revision of verrucaria s.l. (verrucariaceae) in poland. cracow: w. szafer institute of botany, polish academy of sciences; 2012. (polish botanical studies; vol 27). 12. krzewicka b, hachułka m. new and interesting records of freshwater verrucaria in central poland. acta mycol. 2008;43(1):91–98. http://dx.doi.org/10.5586/am.2008.011 13. bielczyk u, editor. the lichens and allied fungi of the polish western carpathians – an annotated checklist. cracow: w. szafer institute of botany, polish academy of sciences; 2003. p. 23–232 (biodiversity of polish carpathians; vol 1). 14. hachułka m. freshwater lichens on submerged stones and alder roots in the polish lowland. acta mycol. 2011;46(2):233–244. http://dx.doi.org/10.5586/am.2011.016 15. ceynowa-giełdon m, adamska a. notes on the genus thelidium (verrucariaceae, lichenized ascomycota) in the kujawy region (north-central poland). ecological questions. 2014;19:25–33. http://dx.doi.org/10.12775/eq.2014.002 16. fałtynowicz w. the lichens, lichenicolous and allied fungi of poland. an annotated checklist. cracow: w. szafer institute of botany, polish academy of sciences; 2003. (biodiversity of poland; vol 6). 17. acharius e. synopsis methodica lichenum, sistens omnes hujus ordinis naturalis detectas plantas, quas, secundum genera, species et varietates disposuit, characteribus et differentiis emendatis definivit, nec non synonymis et observationibus selectis illustravit auctor. lundae: svanborg et soc.; 1814. 18. orange a. four new species of verrucaria (verrucariaceae, lichenized ascomycota) from freshwater habitats in europe. lichenologist. 2013;45(3):305–322. http://dx.doi. org/10.1017/s0024282912000898 19. thüs h. taxonomie, verbreitung und ökologie silicoler süßwasserflechten im außeralpinen mitteleuropa. berlin: j. cramer; 2002 (bibliotheca lichenologica; vol 83). http://dx.doi.org/10.1016/j.scitotenv.2009.06.012 http://dx.doi.org/10.5586/am.2008.011 http://dx.doi.org/10.5586/am.2011.016 http://dx.doi.org/10.12775/eq.2014.002 http://dx.doi.org/10.1017/s0024282912000898 http://dx.doi.org/10.1017/s0024282912000898 abstract introduction material and methods results and discussion hydropunctaria rheitrophila (zschacke) keller, gueidan & thüs verrucaria rheitrophila zschacke thelidium aquaticum servít thelidium minutulum körb. thelidium acrotellum arnold thelidium zwackhii (hepp) a. massal. verrucaria dolosa hepp verrucaria elaeina borrer verrucaria guestphalica auct. verrucaria elaeomelaena (a. massal.) arnold verrucaria hydrophila orange verrucaria denudata zschacke nom. illeg., non nyl. (1858)verrucaria hyd verrucaria latebrosa körb. verrucaria submersella servít references 2015-08-02t16:20:55+0100 piotr otręba new and noteworthy species of lichens from the augustów forest (northeastern poland) 1 of 7published by polish botanical society acta mycologica short communication new and noteworthy species of lichens from the augustów forest (northeastern poland) anna matwiejuk* department of plant ecology, institute of biology, university of bialystok, konstantego ciołkowskiego 1j, 15-245 bialystok, poland * email: matwiej@uwb.edu.pl abstract the augustów forest is one of the biggest forest complex in poland. in this paper, 13 rare species of lichens from augustów forest are presented. four of these species are new to augustów forest: bacidina egenula, lecanora persimilis, rhizocarpon reductum, scoliciosporum pruinosum and one species, rhizocarpon hochstetteri, is new to northeastern poland. short notes on their features and distributions are provided. keywords biodiversity; lichenized fungi; new records; distribution introduction one of the largest forest complexes in poland is the augustów forest, situated in northeastern part of the country, in the podlaskie province (fig. 1). along with the part of forest in lithuania and belarus, which are its immediate neighbors, the augustów forest covers the area over 1600 km2 and thus constitutes one of the largest dense forest complex in europe. the polish part of the augustów forest covers around 1100 km2. in 1989, the northern part of the augustów forest has been appointed wigry national park [1]. in terms of weather condition, the augustów forest is characterized by distinct features of the continental climate. the main stand types of the augustów forest are: fresh coniferous forest, the fresh mixed coniferous forest, and fresh mixed forest, which occupy 39.3%, 27.2%, and 10.3%, respectively, of the area of augustów forest. the main forest-forming species in the augustów forest is sots pine (pinus sylvestris) [1,2]. the augustów forest is characterized by a high diversity of lichen species. currently, in augustów forest (including the wigry national park) there are 365 known lichens taxa [1–3]. in the last decade, intensity of research on the distribution of lichens in poland increased noticeably, using new methods and technologies. as a result, many new stands of different lichen species have been described [4–20], including new stands from augustów forest. the aim of this study was to provide further data on rare and otherwise noteworthy lichen species from the augustów forest. material and methods the specimens for this study were collected during field survey in the augustów forest in 2015 and 2016. doi: 10.5586/am.1102 publication history received: 2017-05-25 accepted: 2017-12-18 published: 2018-01-24 handling editor maria rudawska, institute of dendrology, polish academy of sciences, poland funding the study was supported by the statutory fund of the institute of biology, university of bialystok. competing interests no competing interests have been declared. copyright notice © the author(s) 2018. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation matwiejuk a. new and noteworthy species of lichens from the augustów forest (northeastern poland). acta mycol. 2017;52(2):1102. https:// doi.org/10.5586/am.1102 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:matwiej%40uwb.edu.pl?subject=new%20and%20noteworthy%20species%20of%20lichens%20from%20the%20august%c3%b3w%20forest%20%28northeastern%20poland%29 https://doi.org/10.5586/am.1102 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1102 https://doi.org/10.5586/am.1102 2 of 7© the author(s) 2018 published by polish botanical society acta mycol 52(2):1102 matwiejuk / new and noteworthy species of lichens from the augustów forest morphological characters (thickness, structure, and color of thallus and prothallus, color, shape, and size of soralia and apothecia) were examined using stereomicroscope leica ez4. examination of anatomical features, i.e., thickness and color of hypothecium, hymenium, excipulum proprium, number of spores in asci, size and shape of ascospores were done with light microscope leica dm500. the chemical analyses were carried out using thin layer chromatography (tlc) according to orange et al. [21]. the lichen species nomenclature follows fałtynowicz and kossowska [22]. all recorded sites are located in the atpol grid square system [2]. the geographical coordinates of study sites of the augustów forest were recorded by gps using the wgs84 datum. the collected specimens are deposited in the herbarium of institute of biology, university of bialystok for future reference. lichens were collected in five sites in augustów forest: ■ bg-20, augustów plain mesoregion, augustów forest, the augustów canal, swoboda lock, 53°52'00.2" n / 23°08'41.6" e ■ bg-30, augustów plain mesoregion, augustów forest, the augustów canal, czarny bród, 53°52'49.7" n / 23°12'20.0" e ■ bg-40, biebrza plain mesoregion, augustów forest, augustów forest district, kozi rynek reserve, forest section 53°48' n / 23°13' e ■ bg-21, augustów plain mesoregion, augustów forest, frącki, 53°58'47" n / 23°18'05" e ■ bg-31, augustów plain mesoregion, augustów forest, the augustów canal, perkuć lock, 53°53'56.76" n / 23°19'07.81" e. list of recorded species anisomeridium polypori (m. b. ellis & everh.) m. e. barr in augustów forest, it was previously recorded only by czyżewska et al. [6]. anisomeridium polypori is a nearly cosmopolitan species with a broad ecological amplitude [23]. specimens examined. bg-20, on bark of salix alba, leg. a. matwiejuk. narew river puszcza augustowska bi eb rza ri ve r fig. 1 location of the augustów forest. 3 of 7© the author(s) 2018 published by polish botanical society acta mycol 52(2):1102 matwiejuk / new and noteworthy species of lichens from the augustów forest arthonia muscigena th. fr. the second record in the augustów forest. it is a suboceanic species [6]. arthonia muscigena is specialized to grow on mosses. specimens examined. bg-40, on epiphytic mosses on bark of alnus glutinosa, 2015, 2016, leg. a. matwiejuk. bacidina egenula (nyl.) vĕzda new to the augustów forest. the species was reported for the first time from polish lowland from the białowieża forest [19]. so far, the species has been reported only from the southern part of the country (see fałtynowicz [24]). bacidia egenula is a species often recorded on anthropogenic (concrete) and siliceous substrates (pebbles, stones). specimens examined. bg-20, concrete, 2015, leg. a. matwiejuk. lecanora albella (pers.) ach. so far, it has been recorded from the augustów forest, only from starożyn nature reserve [3]. the species is widespread in temperate to boreal regions of the holarctic, including africa, asia, europe, and north america. it commonly grows on the smooth bark of trees, on which the thallus forms roundish patches [2]. specimens examined. bg-20, on bark of alnus glutinosa, salix alba, 2015, leg. a. matwiejuk. lecanora persimilis (th. fr.) nyl. the species is new to the augustów forest. it has been overlooked for a long time in poland, and was only recently rediscovered after ca. 40 years [19,24–27]. lecanora persimilis is perhaps a boreal to mainly temperate element [27]. it grows on twigs and small branches of neutral-barked deciduous shrubs and trees, especially ash fraxinus, oak quercus [2]. the taxon is endangered in poland, in data deficient category (dd) [28]. specimens examined. bg-20, on fallen branches of quercus robur, 2015, leg. a. matwiejuk. lichenomphalia umbellifera (l.) redhead & al. the second record in the augustów forest [6]. the species is one of few lichens in poland in which mycobiont is basidiomycota. so far, it has been recorded from northeastern poland from several localities [6]. the species occurs on old, decaying logs and on moss-covered soil [2]. lichenomphalia umbellifera is a near threatened species (nt) in poland [28]. specimens examined. bg-40, on decaying wood, 2016, leg. a. matwiejuk. lobaria pulmonaria (l.) hoffm. in the augustów forest, this lichen was reported from several localities [2,3,7,10,29], including wigry national park [1,30–32]. the species was reported first time in kozi rynek reserve. lobaria pulmonaria is widely used as an indicator species of undisturbed old-growth forests [2]. it is an endangered species (en) in poland [28]. specimens examined. bg-40, on bark of fraxinus excelsior, 2016, leg. a. matwiejuk. melanohalea olivacea (l.) o. blanco & al. 4 of 7© the author(s) 2018 published by polish botanical society acta mycol 52(2):1102 matwiejuk / new and noteworthy species of lichens from the augustów forest so far, it has been recorded from the augustów forest only from starożyn nature reserve [3]. melanohalea olivacea is perhaps a circumboreal-montane species, most often found on the barks of betula [3]. melanohalea olivacea is a critically endangered species (cr) in poland [28]. specimens examined. bg-30, on bark of betula pendula, 2016, leg. a. matwiejuk. menegazzia terebrata (hoffm.) körb. so far, this species has been reported from only few localities [2,3,29]. in the kozi rynek nature reserve, it was reported only by cieśliński and tobolewski [29] and cieśliński [2]. menegazzia terebrata is an indicator species for old and biologically rich forests in poland [2]. the species occurs in damp shady woods on barks of trees such as alnus, betula, fraxinus, quercus. menegazzia terebrata is a critically endangered species (cr) in poland [28]. specimens examined. bg-40, on bark of fraxinus excelsior, 2016, leg. a. matwiejuk. rhizocarpon hochstetteri (körb.) vain. this species is characterized by crustose thallus, thin, areolate, concentrated, grey to brown (fig. 2). areoles – 0.8 mm in diameter. prothallus black. apothecia up to 1.3 mm in diameter, insignificantly outstanding over thallus, black, matt. disc nude, flat to easily convex, smooth. margin enough thick or thin, persistent or dying out. epihymenium brown. hymenium 100–110 µm high, hyaline. hypothecium dark brown (fig. 3). asci eight-spored. ascospores 1-septate, hyaline, 10–28 × 6–9 µm. new to the northeastern poland and the augustów forest. in poland, it was reported from only few localities, mainly from the southern part of poland (see fałtynowicz [24]). the species is known from belarus, grodno region [33]. rhizocarpon hochstetteri has a circumboreal/arctic distribution [34]. it grows on either acid or calcareous rocks, stones. rhizocarpon hochstetteri is an endangered species (en) in poland [28]. specimens examined. bg-21, on stone, 2015, leg. a. matwiejuk. rhizocarpon reductum th. fr. new to the augustów forest. this saxicolous lichen is quite common in poland [35]. the species grows on stones and pebbles. the distinction of r. reductum is related to the changes in the taxonomy of rhizocarpon obscuratum (ach.) massal. complex, with hyaline and muriform ascospores [35–37]. examination of the specimens upon which r. obscuratum is based revealed that they are mostly referable to the species currently known as r. lavatum. according to fryday [36], r. reductum is resurrected for specimens with small ascospores and a thallus containing stictic acid formerly placed there, whereas specimens with larger ascospores and a thallus lacking lichen substances are more referred to r. lavatum [35,37]. specimens examined. bg-20, on stone, 2015, leg. a. matwiejuk. fig. 2 rhizocarpon hochstetteri, thallus and apothecia, photo by a. matwiejuk. fig. 3 cross section through the apothecium of rhizocarpon hochstetteri. photo by a. matwiejuk. 5 of 7© the author(s) 2018 published by polish botanical society acta mycol 52(2):1102 matwiejuk / new and noteworthy species of lichens from the augustów forest scoliciosporum pruinosum (p. james) vĕzda new to the augustów forest. in poland, it was reported only from few localities, in old deciduous forests (see fałtynowicz [24]). it is a mild-temperature species, grown on barks of deciduous trees. scoliciosporum pruinosum is an endangered in poland, in data deficient category (dd) [28]. specimens examined. bg-40, on bark of alnus glutinosa, 2016, leg. a. matwiejuk. thelotrema lepadinum (ach.) ach. in the augustów forest, it was reported from only few localities – starożyn, perkuć, kozi rynek nature reserves [2,3,6,29]. this is an indicator species for old and biologically rich forests in poland [2]. it commonly grows on sheltered, smooth-barked, deciduous trees. thelotrema lepadinum is an endangered species (en) in poland [28]. specimens examined. bg-40, on barks of alnus glutinosa, fraxinus excelsior, quercus robur, 2016, leg. a. matwiejuk. in conclusion, four species of lichens, namely bacidina egenula, lecanora persimilis, rhizocarpon reductum, scoliciosporum pruinosum, presented in this paper are new species for the augustów forest, whereas rhizocarpon hochstetteri is new to northeastern poland. acknowledgments i am grateful to the anonymous reviewers for helpful comments and suggestions. references 1. fałtynowicz w. porosty wigierskiego parku narodowego. parki narodowe i rezerwaty przyrody. 1994;13(3):9–28. 2. cieśliński s. atlas rozmieszczenia porostów (lichenes) w polsce północno-wschodniej. phytocoenosis. supplementum cartographiae geobotanicae. 2003;15:1–426. 3. zielińska j. porosty rezerwatu starożyn. acta mycol. 1969;5:135–148. https://doi.org/10.5586/am.1969.012 4. kukwa m. lepraria incana (l.) ach. in: bielczyk u, cieśliński s, fałtynowicz w, editors. atlas of the geographical distribution of lichens in poland. p. 4. cracow: w. szafer institute of botany, polish academy of sciences; 2004. p. 45–57. 5. kukwa m. lecanora thysanophora (lecanoraceae, zlichenizowane ascomycota) w polsce. fragm florist geobot pol. 2005;12(2):385–391. 6. czyżewska k, motiejūnaitė j, cieśliński s. new and noteworthy species of lichens and allied fungi from north-eastern poland. acta mycol. 2005;40(2):277–291. https://doi.org/10.5586/am.2005.025 7. ryś a. granicznik płucnik lobaria pulmonaria w lasach państwowych i jego ochrona. olsztyn: studio avalon; 2005. 8. motiejūnaitė j, czyżewska k. additions to the biota of lichens and lichenicolous fungi of poland with note on lecania prasinoides in eastern and central europe. pol bot j. 2008;3(2):155–162. 9. syrek m, kukwa m. taxonomy of the lichen cladonia rei and its status in poland. biologia. 2008;63(4):493–497. 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(acta botanica silesiaca, monographiae; vol 8). 23. aptroot a. notes on taxonomy, distribution and ecology of anisomeridium polypori. lichenologist. 1999;31(6):641–642. https://doi.org/10.1006/lich.1999.0233 24. fałtynowicz w. the lichens, lichenicolous and allied fungi of poland. an annotated checklist. cracow: w. szafer institute of botany, polish academy of sciences; 2003. 25. kubiak d. lichens and lichenicolous fungi of olsztyn town (ne poland). acta mycol. 2005;40(2):293–332. https://doi.org/10.5586/am.2005.026 26. łubek a. antropogeniczne przemiany bioty porostów świętokrzyskiego parku narodowego i otuliny. fragm florist geobot pol. 2007;10:3–94. 27. śliwa l. a revision of the lecanora dispersa complex in north america. pol bot j. 2007;52(1):1–70. 28. cieśliński s, czyżewska k, fabiszewski j. red list of the lichens in poland. in: mirek z, zarzycki k, wojewoda w, szeląg z, editors. red list of plants and fungi in poland. cracow: w. szafer institute of botany, polish academy of sciences; 2006. p. 73–89. 29. cieśliński s, tobolewski z. porosty polski północno-wschodniej. i. acta mycol. 1989;27:57–100. https://doi.org/10.5586/am.1989.003 30. bystrek j, matwiejuk a. porosty rezerwatu monkinie w wigierskim parku narodowym. annales universitatis mariae curie-sklodowska. sectio c, biologia. 1994;49(3):31–42. 31. bystrek j, przepiórkowska a. porosty rezerwatu sernetki w wigierskim parku narodowym. annales universitatis mariae curie-sklodowska. sectio c, biologia. 1994;49(4):43–58. 32. bystrek j, matwiejuk a. porosty obszarów chronionych i proponowanych do ochrony w lasach wigierskich. annales universitatis mariae curie-sklodowska. sectio c, biologia. 1999,54:93–124. 33. golubkov v, matwiejuk a. some new records of rhizocarpon from northeastern poland and north-western belarus. acta mycol. 2009;44(2):201–210. https://doi.org/10.5586/am.2009.018 34. fryday am. a revision of the species of the rhiyocarpon hochstetteri group occurring in the british isles. lichenologist. 2002;34(6):451–477. https://doi.org/10.1006/lich.2002.0416 35. matwiejuk a. rhizocarpon lavatum and r. reductum (rhizocarpaceae, ascomycota), two misunderstood taxa found in the gorce mts (polish carpathians). acta mycol. 2012;47(1):121–126. https://doi.org/10.5586/am.2012.013 https://doi.org/10.5586/am.2010.028 https://doi.org/10.5586/am.1087 https://doi.org/10.1006/lich.1999.0233 https://doi.org/10.5586/am.2005.026 https://doi.org/10.5586/am.1989.003 https://doi.org/10.5586/am.2009.018 https://doi.org/10.1006/lich.2002.0416 https://doi.org/10.5586/am.2012.013 7 of 7© the author(s) 2018 published by polish botanical society acta mycol 52(2):1102 matwiejuk / new and noteworthy species of lichens from the augustów forest 36. fryday am. on rhizocarpon obscuratum (ach.) massal., with notes on some related species in the british isles. lichenologist. 2000;32:207–224. https://doi.org/10.1006/lich.2000.0269 37. ihlen pg. taxonomy of the non-yellow species of rhizocarpon (rhizocarpaceae, lichenized ascomycota) in the nordic countries, with hyaline and muriform ascospores. mycol res. 2004;108:533–570. https://doi.org/10.1017/s0953756204009803 https://doi.org/10.1006/lich.2000.0269 https://doi.org/10.1017/s0953756204009803 abstract introduction material and methods list of recorded species acknowledgments references 2018-01-24t10:39:29+0000 piotr otręba strigolactones as mediators between fungi and plants 1 of 8published by polish botanical society acta mycologica review strigolactones as mediators between fungi and plants anita kowalczyk1,2, katarzyna hrynkiewicz1,2* 1 department of microbiology, faculty of biology and environmental protection, nicolaus copernicus university in toruń, lwowska 1, 87-100 toruń, poland 2 center of modern interdisciplinary technologies, nicolaus copernicus university in toruń, wileńska 4, 87-100 toruń, poland * corresponding author. email: hrynk@umk.pl abstract a constantly changing environment is challenging for all organisms on earth, especially for terrestrial plants, which face several environmental stresses despite their static way of life. in attempts to understand the mechanisms responsible for plant growth and development, scientists have recently focused on a small group of carotenoid derivatives called “strigolactones” (sls), which are synthesized mostly in the roots in response to a variety of external factors. strigolactones are compounds that define plant plasticity towards many environmental factors, including the establishment of mycorrhizal symbiosis under nutrient-deficient conditions. as exogenous signals, they can stimulate the branching of arbuscular mycorrhizal fungal (amf) hyphae and as endogenous signals they adjust a plant architecture, including changes within the roots, allowing host plant and fungi to meet. sls can also function as signaling molecules that allow colonization and establishment of the later stages of mutualistic symbioses between organisms such as amf. sls act on amf metabolism by stimulating its mitochondrial respiration. genes encoding enzymes crucial for sl biosynthesis – ccd7 and ccd8 – are also found in gymnosperm genomes, which encourages speculation that strigolactones may also be part of a host-plant and ectomycorrhizal fungi signaling pathway during the establishment of symbiosis. nevertheless, sls impact on ectomycorrhiza formation remain unknown. the broad spectrum of sl bioactivity has made these compounds valuable from an industrial perspective. in the future, sls may be commercialized in plant protection products, biostimulants, or as substances used in genetic engineering to allow the creation of crops capable of growing under disadvantageous conditions. keywords plant–microbial interactions; symbiosis; arbuscular mycorrhizal fungal (amf); ectomycorrhizal fungi (ecm); pathogenic fungi strigolactones (sls): a new class of plant hormones strigolactones (sls) are evolutionarily old signaling molecules that affect plant growth and development. strigolactones belong to a group of naturally synthesized secondary metabolites with a butenolide ring in their structure (called the d-ring) connected by an ether enol bridge to a second moiety [1,2]. the first indications of the presence of sls were found while studying the interactions between parasitic plants and their hosts. it was suggested that there may exist a factor that has the ability to stimulate the germination of parasitic plants from the genus striga. strigol was isolated for the first time in 1966 from cotton root exudates (gossypium hirsutum l.) [3]. strigolactones participate in the modification of root and shoot architecture and also in the establishment of the symbiosis between plants and microorganisms, e.g., arbuscular mycorrhizal fungi (amf). this is the basis for the inclusion of sls in the phytohormone family. sls have doi: 10.5586/am.1110 publication history received: 2018-04-09 accepted: 2018-06-19 published: 2018-09-24 handling editor dorota hilszczańska, forest research institute, poland authors’ contributions ak wrote the manuscript; kh supervised this work funding this research was financially supported by a grant from the national science center (poland) (2016/23/b/ nz9/03417). competing interests no competing interests have been declared. copyright notice © the author(s) 2018. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation kowalczyk a, hrynkiewicz k. strigolactones as mediators between fungi and plants. acta mycol. 2018;53(2):1110. https:// doi.org/10.5586/am.1110 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:hrynk%40umk.pl?subject=strigolactones%20as%20mediators%20between%20fungi%20and%20plants https://doi.org/10.5586/am.1110 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1110 https://doi.org/10.5586/am.1110 2 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1110 kowalczyk and hrynkiewicz / strigolactones as mediators between fungi and plants recently been shown to be active in the following processes: (i) photomorphogenesis, secondary growth, (ii) leaf senescence, (iii) and plant response to stress factors, such as a lack of nutrients or water [4–6]. the wide range of plant–sl interactions, both systemic and environmental, is the reason for the growing scientific and commercial interest. in the future, sls may be used in plant protection products, e.g., herbicides, or as biostimulators that can boost the efficiency of nutrient intake [7]. the role of strigolactones in symbiotic interactions between arbuscular fungi (amf) and plants mycorrhiza is a widespread phenomenon in the plant kingdom, which relates to more than 90% of terrestrial plant species. the formation of symbiotic radical associations is extremely significant for the physiology of both symbionts and ecosystem stability, which depends on the functionality of the symbiosis [8]. establishment of arbuscular and ectomycorrhizal symbiosis in plant roots may involve the same factors regulating the recognition of symbionts and further symbiotic actions. such factors may undoubtedly include strigolactones because the genes responsible for their biosynthesis have been shown to be present in almost all plants that exist in mutualistic relationships with fungi. currently, research on the secretion of strigolactones is far more advanced in the case of arbuscular mycorrhiza. the main reason for this is the numerous similarities between the early stages of arbuscular mycorrhiza development and the symbiosis between rhizobium bacteria and legumes. the following subsections will present the current knowledge on arbuscular and ectomycorrhizal systems and their sl synthesis activity. the branching of amf hyphae towards the roots of its host is considered to be the most important stage in the entire formation of arbuscular mycorrhizae. how does it happen and what kind of role do strigolactones play when released into the rhizosphere? amf spores can germinate in the soil even when no potential host is nearby, but germinated spores die in the absence of root exudates. this leads to the conclusion that there must be some kind of presymbiotic communication among the symbionts, as is the case in the rhizobium–legume symbiosis, where flavonoids released by the plants are the signal for the transcription of genes vital for the production of bacterial nod factors. legume plants are also hosts for amf, and becard et al. [9] conducted an experiment in 1995 in which they studied maize mutants that were unable to produce flavonoids. the authors used exudates from transformed carrot roots (because the carrot was the model plant used in the fungal interaction studies) that induced branching in both mutated and wild-type maize at the same level [9]. before sls earned their name, they were called branching factors (bfs). in 2005, akiyama et al. [10] identified a compound called 5-deoxystrigol in lotus japonicus exudates, which is similar to other natural sls (sorgolactone, strigol, or synthetic sl gr24) caused intensive hyphal branching of gigaspora margarita, a fungus belonging to the amf family. furthermore, the characteristics of 5-deoxystrigol conformed with the criteria of the nagahashi and douds [11] biotest for chemical analysis and identification of sls. in 2003, tamasloukht et al. [12] used partially purified fractions of carrot exudates to show that sls could induce the expression of mitochondrial fungal genes, along with an intense stimulation of mitochondrial respiration and effects on mitochondrial reorganization prior to the onset of branching. three years later, besserer et al. [13] applied the synthetic analogue of sl – gr24 – and noticed an intense and sudden increase in cell proliferation and changes in the shape and amount of mitochondria. subsequently, besserer et al. [14] and besserer and roux [15] demonstrated that these effects were accompanied by a rapid increase in cell respiration in mitochondria, increased nadh concentration, and an increase in atp levels in the fungal cells. this kind of mitochondrial activation could lead to the oxygenation of lipids, which are the carbon source found in the amf spore reserve materials. thus, sls may be the key components in root exudates that are capable of triggering lipid catabolism in the presymbiotic stage of plant–fungi interactions. the data discussed above support the hypothesis that sls participate in specific mechanisms of recognition and signal transduction between the host plant and amf [16]. 3 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1110 kowalczyk and hrynkiewicz / strigolactones as mediators between fungi and plants strigolactones play a crucial role in amf root colonization processes. they are involved in the recognition stage, thus increasing the probability of encountering a root zone that is well adapted to colonization [17]. inoculation of pea mutants that were unable to produce sls with amf spores exhibited significantly lower levels of root colonization compared to that of wild-type plants. gr24 application partially reversed the observed effect [18]. these studies provide the first direct evidence of the essential role that sls play in the process of hyphae branching, as well as host root colonization. nevertheless, as garcía-garrido et al. [19] rightly pointed out, in the case of mutated plants, that mycorrhization should not occur at all. however, if it takes place, it may indicate that the mutants have a reduced level of sl synthesis, or that these compounds have the ability to stimulate amf branching and root colonization and the plant can produce other substances that control the mechanisms allowing interaction between the symbionts. in addition, amf spores can germinate independently, and therefore symbiosis may form in the vicinity of the roots. the effect of gr24 is enigmatic and it is difficult to determine whether it is the result of induction of spore germination or stimulation of hyphae branching. mutated plants inoculated with gr24 should display a profuse branched fungi phenotype, regardless of the distance between the hyphae and the roots [19]. hence, an explanation for the mechanisms involved in the interaction of sls with amf in the presymbiotic and colonization stage is necessary to answer these emerging questions. an interesting issue of the previously mentioned study area was the discovery of the diffusible factor from amf, called myc factor (or myc-lco). myc factors are lipochitooligosaccharides that are able to activate the early plant response to the following mycorrhization (fig. 1) through the induction of gene expression related to the transduction pathway and biogenesis of the plant’s prepenetration apparatus (ppa) allowing fungi to grow inside the host cells [20]. despite the suggestions emerging from various studies, the myc factor transduction is independent of nod factor transduction. kosuta et al. [21] conducted a study in which they separated gigaspora rosea spores and transformed medicago truncatula roots (a plant that has the ability to establish symbiosis with both mycorrhizal fungi and rhizobia) with a cellophane membrane that allowed their growth in proximity and signal exchange. the fungal mycelium of m. truncatula secreted substances smaller than 3.5 kda that activated expression of the mtenod11 gene. the expression was also synchronized with the induction of hyphae branching. this result could not be observed in the dead spores of amf or mycelium of pathogenic strains. another factor confirming this pathway’s independence was that m. truncatula mutants, unable to form a mycorrhizal or nodule symbiosis, begin the wild-type mtenod11 activation when exposed to the amf [22]. the main challenge of contemporary research is to identify the myc factors, recognize their signaling pathways, and distinguish plant responses. the existence of myc factors is not the only indicator of amf and rhizobia interactions with plants. in 2000, vierheilig et al. [23] demonstrated the existence of a plant regulatory mechanism called autoregulation of mycorrhization, which inhibits mycorrhization after a certain level is reached, which was similar to the autoregulation of nodulation (aon) that prevents nodule overgrowth in roots. autoregulation of mycorrhization is not related to phosphorus availability [24] or to carbon-access competition [25]. the data collected thus far indicate that sl levels in mycorrhizal plants vary over time. according to the lendzemo and kuyper [26] research, plants such as sorgo and maize show a high tolerance to plant–parasite infections compared to nonmycorrhizal control plants, which may be associated with the aforementioned autoregulation. it also may be related to the reduced stimulation of parasitic plant germination as a result of reduced sl secretion into the rhizosphere. thus, mycorrhization can negatively regulate sl production, affecting the branching processes and general progress of amf colonization. although only the sls effect on autoregulation of mycorrhization has been documented, it may not be the only plant mechanism to prevent amf colonization outgrowth that could cause the mutualistic relation to turn parasitic [19]. 4 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1110 kowalczyk and hrynkiewicz / strigolactones as mediators between fungi and plants strigolactones and ectomycorrhizal fungi (ecm) although there are seven types of mycorrhizal [27] associations, it is the previously mentioned arbuscular mycorrhiza (endomycorrhiza) and ectomycorrhiza that are considered the most ecologically and economically important. however, the arbuscular mycorrhiza is the most widely studied type and consequently, the mechanisms associated with arbuscular mycorrhiza establishment in host plants and fungi are far better understood than those of ecm symbiosis. the initial stage of ectomycorrhiza formation includes a signal exchange between the two symbionts. plants secrete primary and secondary metabolites, along with sugars, hormones, and enzymes that can affect the root microbiome. in 1987, fries et al. [28] showed that abietic acid present in pinus sylvestris root exudates stimulated suillus spp. spore germination. the molecules responsible were flavonoids – rutin secreted by eucalyptus globulus spp. bisocata stimulated the growth of pisolithus spp. hyphae [29]. when data confirming the positive activity of sls towards amf was published [10], researchers realized a new possibility regarding the fig. 1 schematic summary of the possible root–fungi interactions involving sls. (a) ectomycorrhiza. the initial stage of ecm symbiosis establishment is the signal exchange between the host and the fungi. plants secrete a variety of substances with the flavonoids responsible for the stimulation of spore germination [29] and fungi release of the still poorly studied lipochitooligosaccharides (lco), chitooligosaccharides (co), and small secreted proteins (ssp). the current lack of research conducted on the sl–ecf relationship makes it impossible to assume with certainty that sls are not involved in the signaling mechanisms, as they can act on pathways other than recognition symbiosis pathways or interact with fungal effectors [31]. (b) arbuscular mycorrhiza. sls are proven to act in the presymbiotic stage as triggering spore-germination signals when released into the soil in the proximity of amf spores. secreted sls along with other plant exudates induce the expression of fungal mitochondrial genes followed by an intense stimulation of mitochondrial respiration with increased nadh and atp concentration in the fungal cells [12,14,15]. this is the proposed way in which lipid catabolism allowing spores to germinate is activated through host–fungi signaling. after germination, sls allow fungal hyphae to branch towards the roots prior to the proper recognition of symbionts, leading to the activation of the fungal signaling pathway and the release of myc lipochitooligosaccharides (myclcos). subsequently, myclcos induce the expression of nod genes that is correlated with the induction of hyphae branching [22]. (c) pathogenic fungi interactions. in the preliminary data, sls were not found to induce hyphal branching or respiration processes of the examined plant pathogens [30,34]. even though there is no clear evidence that sls are or are not a part of a host–pathogen signaling relation, it is an area that is interesting and undoubtedly tempting to explore, with some indications that sls may be engaged in this process as signals preventing infection by promoting am symbiosis. 5 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1110 kowalczyk and hrynkiewicz / strigolactones as mediators between fungi and plants possible roles of sls with respect to both types of mycorrhiza. steinkellner et al. [30] conducted a study that included the ectomycorrhizal fungi paxillus involutus, laccaria bicolor, amanita muscaria, and cenococcum geophilum. none of these expressed any changes in branching patterns after the application of exogenous gr24. nevertheless, this preliminary data should be confirmed. furthermore, sls may activate symbiosis pathways or positively affect the production of effectors or fungal chitin signals (fig. 1) [31]. there are still many questions regarding whether fungal compounds, such as lipochitooligosaccharides (lco), chitooligosaccharides (cos), small secreted protein (ssp), and plant hormones, affect the early stages of ecm symbiosis formation [32]. sls are found in almost all terrestrial plants, they are synthesized at lower concentrations even in nonhost plants, and ccd7 and ccd8 enzymes were found to play a key role in sl biosynthesis in genomes and transcriptomes of gymnosperms. consequently, research on the relationship between ecm symbiosis and strigolactone should be continued. strigolactones and pathogenic fungi in the section focused on amf it was stated that the stimulating effect of sls on amf spore germination has been documented, but do sls work in a similar way for plant pathogens? in research by steinkellner et al. [30], the authors sought to establish whether the sl synthetic analogue gr24 could stimulate the microconidia germination of fusarium oxysporum f. sp. lycopersici. the results were negative, suggesting that sls may act as specific signals for amf. gr24 effects on hyphal branching patterns were also examined for soil-borne pathogens (rhizoctonia solani, fusarium oxysporum, verticillum dahlia) and aerial plant-part pathogens (botrytis cinerea, cladosporium sp.) with the same result, indicating no effects on their growth [30]. another study conducted in 2001 by martinez et al. [33] showed that fractions of maize root exudates could induce the transition of yeast to a hyphal form. in later research, sabbagh [34] analyzed the transcriptome expression of a haploid strain of s. reilianum using different concentrations of gr24, and most of the expressed sequence tag (est) after sl application was related to genes taking part in cell respiration processes. exogenous gr24 effects on ustilago maydis, a fungus causing smut on maize, induced genes involved in cell respiration at 1 h, increased the process at 5 h, and stopped it at 8 h postsupplementation; further analysis of cells induced at 8 h showed the lack of any candidate gene transcripts [34]. even if strigolactones do not affect the pathogenicity pathway of other fungi, they can still be used as compounds to prevent host plants from being infected through the promotion of arbuscular mycorrhiza formation. possible uses of strigolactones in industry sls are characterized by high activity and effectiveness in biological systems. thus, research into their potential use is being intensified. sls may be used as biofertilizers thereby indirectly promoting the initiation and establishment of amf symbiosis or rhizobium–legume symbiosis because microorganism associations are the primary way plants cope with a lack of nutrition [7]. however, there are numerous problems standing in the way of commercialized products based on sl bioactivity. high sl production costs exist primarily because the method of their laboratory synthesis has not yet been optimized. the high lability of sls hampers the processes to isolate and purify them. the isolation is exceptionally hard to perform because the natural synthesis of sl is very limited. for example, cotton synthesizing two naturally occurring strigolactones – strigol and strigol acetate – can secrete 15 and 2 pg/plant/day, respectively [35]. consequently, their mass production would require appropriate laboratory equipment and a specially developed methodology. attempts to accumulate sls in a culture medium failed because of their rapid degradation. strigol and its analogues are highly susceptible to hydrolysis in alkaline media because of the high reactivity of their enol ether bridge. in the most recent studies, a mixture of a synthetic sl analogue – gr24 – can last up to 10 days in a 6 of 8© the author(s) 2018 published by polish botanical society acta mycol 53(2):1110 kowalczyk and hrynkiewicz / strigolactones as mediators between fungi and plants neutral ph, whereas 5-deoxystrigol only lasts 1.5 days. the stability of sls in the soil is one of the most important aspects of their future use because of the ddt effect, which is known to this day. on the other hand, their chemical structure needs to be refined, so it could prevent their rapid hydrolysis. therefore, plant cultures are not the most efficient method for sl acquisition and organic synthesis is complicated because of the lack of knowledge regarding some of the natural biosynthesis stages of sls [7]. there are still many unknowns that need to be identified before we will be able to synthesize sl products for the market. summary plants exhibit a high degree of plasticity towards environmental conditions. sls may be molecules that play a key role in plant reactions because they act as (i) exogenous signals perceived by microorganisms in their vicinity and (ii) endogenous compounds regulating the shoot and root architecture according to plant needs. in both cases, the sl signaling pathways are a response to environmental factors, such as nutrient availability, temperature, light, or biotic stress. interactions occurring with sls and other phytohormones that allow plants to respond correctly have yet to be investigated at the cellular and molecular level. one very interesting yet poorly understood issue is that of sl interactions with microorganisms other than amf. the predicted ubiquity of sls in a variety of land plants and numerous questions 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https://doi.org/10.1271/bbb.69.98 abstract strigolactones (sls): a new class of plant hormones the role of strigolactones in symbiotic interactions between arbuscular fungi (amf) and plants strigolactones and ectomycorrhizal fungi (ecm) strigolactones and pathogenic fungi possible uses of strigolactones in industry summary references 2018-09-24t18:33:34+0100 piotr otręba can the soil geology and chemistry analysis of a site predict the geographic origin of wild edible mushrooms (porcini group)? 1 of 15published by polish botanical society acta mycologica original research paper can the soil geology and chemistry analysis of a site predict the geographic origin of wild edible mushrooms (porcini group)? elia ambrosio1*, pietro marescotti1, gian maria niccolò benucci2, grazia cecchi3, michele brancucci4, mirca zotti3, mauro giorgio mariotti1,5 1 department of earth, the environment and life science (distav), university of genoa, corso europa 26, 16132 genoa, italy 2 department of plant, soil and microbial sciences, michigan state university, 48824 east lansing, mi, usa 3 laboratory of mycology, department of earth, the environment and life science (distav), university of genoa, corso europa 26, 16132 genoa, italy 4 geospectra s.r.l. spin off university of genoa, via palmaria 9/6 l, 16121 genova, italy 5 laboratory of plant biology, department of earth, the environment and life science (distav), university of genoa, corso europa 26, 16132 genoa, italy * corresponding author. email: elia.ambrosio.10@gmail.com abstract this study aimed to assess the element content of porcini mushrooms collected from broadleaf mediterranean forests (nw italy) and underlying soil layers, and to elucidate the chemical connection between the mushrooms and their geographic site of origin. comparing the elements in mushrooms with those in soil samples, we observed that the concentration of some microelements detected in mushrooms had similar distribution as that measured in both the soil layers assessed, especially with surface soil. statistical analyses showed that the microelement pattern in mushrooms reflects the soil site of origin. moreover, by comparing our results with other studies, we observed that the soil where porcini grow is characterized by a high concentration of zinc. some toxic elements were also detected in mushroom samples. analysis of elements in mushrooms and soil layers can be used for quality assurance of natural products and help distinguish them from uncertified and unknown-origin products. keywords wild edible mushrooms; boletus edulis group; traceability; soil element content; mushroom safety introduction it is well known that mushrooms are able to accumulate diverse chemical elements from the environment (e.g., air, water, and soil) or substrates (e.g., wood) where they grow [1–4]. although accumulation capability and the presence of chemical elements in sporocarps [5] varies according to nutritional requirements and fungal genotypes, several authors have shown that the element content of mushrooms is mainly influenced by the chemical composition of the growing substratum, specifically the soil [6,7]. despite these results, the majority of studies only analyze the presence of heavy metals inside a few fungal species from both polluted and unpolluted sites [4–8]. to the best of our knowledge, only few studies have analyzed the influence of geology, soil-mineralogy, and soil-chemistry on the element content in sporocarps, as well as the correlation between chemical elements in mushrooms and in the underlying soil layers [9–11]. doi: 10.5586/am.1130 publication history received: 2019-06-15 accepted: 2019-08-29 published: 2019-12-30 handling editor andrzej szczepkowski, faculty of forestry, warsaw university of life sciences – sggw, poland authors’ contributions ea conducted the fieldwork, did the species identification, analyzed the data, and wrote the manuscript; gmnb contributed to the statistical analysis; pm and mb performed the chemical analysis; gc and mz contributed to the fieldwork; mgm coordinated the phd project; ea, pm, and mz contributed equally to the study plan funding this study was carried out in the framework of a phd project in applied botany in agriculture and the environment, university of genoa, italy. ea was supported by miur (ministry of education, universities and research – italy) doctoral fellowship (2012–2014). competing interests no competing interests have been declared. copyright notice © the author(s) 2019. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation ambrosio e, marescotti p, benucci gmn, cecchi g, brancucci m, zotti m, et al. can the soil geology and chemistry analysis of a site predict the geographic origin of wild edible mushrooms (porcini group)? acta mycol. 2019;54(2):1130. https://doi. org/10.5586/am.1130 mailto:elia.ambrosio.10%40gmail.com?subject=can%20the%20soil%20geology%20and%20chemistry%20analysis%20of%20a%20site%20predict%20the%20geographic%20origin%20of%20wild%20edible%20mushrooms%20%28porcini%20group%29? https://doi.org/10.5586/am.1130 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1130 https://doi.org/10.5586/am.1130 2 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers wild edible mushrooms are the most valuable nonwood forest products in the world [12,13]. currently, more than 2,000 fungal species are known to produce edible sporocarps, which are harvested, consumed, and marketed in more than 85 countries [14,15]. current estimations assign edible mushrooms a global market value that is at least $2 billion higher than that of timber products [16–20]. in particular, “porcini” (boletus edulis s. s., a group that also includes the species b. aereus bull., b. pinophilus pilát & dermek, and b. reticulatus schaeff ) are among the most valuable and widelyconsumed groups of wild mushrooms worldwide. from 20,000 to 100,000 metric tons of porcini are estimated to be consumed annually, at fresh-mushroom prices that varied from $60 to $200/kg in 2009 [21,22]. over the last few decades, consumer demand for specialty and high quality agrifood products has grown incredibly, and the market is greatly expanding [14–16]. for these reasons, the concept of traceability, in terms of detecting the origin and quality of products, has become an important criterion in the food sector [23–25]. despite the existence of a legal system [26–28], there is a further need to distinguish between highand low-quality produce and to develop new methods and technologies for food traceability. identifying a product’s geographic site of origin means that we can then identify possible risks to human health. most fraudulent products do not satisfy food safety requirements, and they can profoundly damage confidence in typical food products. molecular analysis of proteins and fatty acid profiles as well as dna barcoding have been widely used to identify the origin of food products [29–33]. although molecular techniques are theoretically informative and precise, they involve high processing costs and do not allow us to clearly distinguish product authenticity, i.e., the geographical site of origin. different chemical and physical approaches, such as gas chromatography, mass spectrometry, nir spectroscopy, and the analysis of trace elements and stable isotopes have been developed as alternatives to establish the geographical origin of agri-food products [34–38]. several authors agree that soil elements can be favorably used as “signatures” of the geographic provenance of a sample [9,10,24]. for instance, nikkarinen and mertanen [9] analyzed the element content in a boletus edulis group and lactarius trivialis (fr.) fr., growing in two different geological regions in finland, to determine whether geochemical fingerprints can be detected in mushrooms. they found that fungal samples differed considerably in trace content based on the geological and geographic site of origin. because soil elements can be used as fingerprints of the geographic provenance of an organic sample, we analyzed the chemical content in mushrooms and soil layers to detect the origin, quality, and authenticity of the group of wild edible mushrooms known as porcini. the aims of this study was (i) to investigate the relationships between the element content of porcini mushrooms and the underlying soil layers; (ii) to clarify the chemical connection between the mushrooms and their geographic site of origin; (iii) to assess the presence of toxic elements in the mushrooms; (iv) to identify possible soil-geochemical markers that can be used to trace porcini mushrooms to their harvesting site; and (v) to chemically characterize the soil of selected porcini sites. material and methods study area this study was carried out in two sites, an oak wood and a beech forest, located in liguria (province of savona) in nw italy (fig. 1). the first site (site 1), near the province of la maddalena (44°30'14" n, 8°29'17" e), covers an area of approximately 7,500 m2 with an altitude ranging from 340 to 380 m a.s.l. the area is dominated by quercus cerris l., and the site is classified as a high forest. the second site (site 2) is located near the province of vereira (44°27'3" n, 8°32'42" e) and covers a total area of approximately 10,000 m2. the altitude is 1,000 m a.s.l. the area is dominated by fagus sylvatica l., and it is classified as a high forest. 3 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers site 1 lies in a geologically complex area characterized by soils deposited on four different rocks: serpentine schists, calceschists, chlorite-actinolite schists, and conglomerates belonging to the tertiary piedmont basin’s stratigraphic succession. in contrast, site 2 lies in a geologically homogeneous area characterized by soils deposited exclusively on calceschist rock. the climate is temperate oceanic sub-mediterranean for both sites [39]. the mean annual temperature is 12°c [from 0°c (min) in january to 25°c (max) in july)]. mean annual rainfall is 912 mm [33 mm (min) in july; 122 mm (max) in october] [40]. sample collection in spring and fall 2014 and 2015, 20 samples of boletus reticulatus schaeff (fig. 2), were collected from the two study sites. the mushroom samples are labeled in tab. 1, tab. 6, and tab. 7 and fig. 5–fig. 7 with the identifiers “m1–m20”. species identification was carried out for both fresh and dried specimens by macro and microscopic observations and a series of monographs and keys [41–43]. nomenclature and author abbreviations were used in accordance with hibbett et al. [44], ima [45], and cbs [46]. two soil samples (1 kg each) were collected beneath each mushroom: one soil sample from the surface layer (from 0 to 20 cm, labeled sl1–sl20), and the other from the deeper layer (from 20 cm to 40 cm, labeled dl1–dl20). to characterize each site from a geochemical and mineralogical perspective, we also randomly collected 10 soil samples (1 kg each) at each site, at a depth between 5 and 20 cm (labeled s1–s20). these soil samples were recorded in the center of 10 circular plots (4-m radius) selected along a transect line as proposed by feest [47]. all soil samples were sieved in situ to remove particles >2 cm. fig. 1 geographic location of the study sites. fig. 2 picture of b. reticulatus schaeff. 4 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers chemical and physical analysis the complete sporocarps (cap and stipe) and soil samples were brushed in the field, washed with distilled water, and dried at 60°c for 48 hours. they were then powdered, sieved and stored in hermetic plastic containers in the laboratory. elements were detected by a field-portable energy dispersive x-ray fluorescence (fp-edxrf) spectrometer (x-met7500; oxford instruments). this is one of the simplest, most accurate, and most economic analytical tools to detect the chemical composition of many mineral and organic substrates [6,25,48–52]. a total of 5 g of each sample was exposed to x-rays and related element concentrations were expressed in ppm. the ph was measured using the wtw multiline p3 set ph meter after equilibrating the soil fraction (<2 mm) in deionized water for 12–16 hours. the soil granulometry (% of clay, silt, and sand) was classified according to folk [53]. statistical analyses for each mushroom and soil sample, descriptive statistics (min, max, mean, standard deviation) were calculated for element concentration, and differences in element content were displayed using boxplots. pearson’s correlation (r) was used to test the hypothesis of linear independence between two variables. this coefficient indicates how well two data sets are interconnected (positively, negatively, or no connection) [54]. in this study, variables were represented by element concentration detected in the sporocarps and in both the soil portions (surface and deep layers). a set of multivariate analyses was used to measure the degree of dissimilarity between mushrooms and soil samples. specifically, the hierarchical cluster analysis (ca) (using the bray–curtis dissimilarity index and unweighted pair group method with arithmetic mean upgma) was performed to discern the geographic site of provenance of the samples using their degree of dissimilarity in chemical composition [54]. principal component analysis (pca) was used to reduce dimensionality and summarize all variables into a few principal components, which explain the greatest amount of variance in the data and can be visualized graphically. pca was calculated in r using factominer and factoextra packages [54,55]. finally, the indicator species analysis (isa) technique was performed on the element concentration matrix in order to identify possible fungal and soil chemical markers [54,55]. before performing statistical analyses, data were normalized using the formula f(x) x/sum(x). data analyses were performed using the r software environment for statistical computing and graphics version 3.5.1 [56]. results element content in mushrooms and soil layers overall, we analyzed the elements in 80 samples: 20 sporocarps (10 recorded at site 1 and 10 at site 2) and underlying soil samples (20 from the surface layer and 20 from the deep soil). in addition, 20 soil samples were taken from a depth between 5 and 20 cm from the top of the soil, to perform a general geochemical characterization of the sites. chemical element concentrations measured in the b. reticulatus samples are summarized in tab. 1. full details on the elements detected in the mushroom samples (m1–m20) are provided in appendix s1. altogether, we detected the presence of 10 different elements: ca, ti, mn, fe, cr, ni, cu, zn, sr, and sb. in some samples, the concentration of certain elements (ti, fe, sr) were below or very close to the detection limit [43 ppm, 10 ppm, and 1 ppm, respectively (appendix s1)]. conversely, high zn concentration was found in most of the analyzed mushroom samples, specifically; 105 and 311 ppm were the maximum values detected at site 1 and site 2, respectively. 5 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers element concentrations measured in soil samples (sl1–sl20 and dl1–dl20), collected beneath each mushroom sample, are summarized in tab. 2 and tab. 3, and detailed concentration values are listed in appendix s2 and appendix s3, respectively. altogether, 26 elements were detected in both the soil layers. in some samples, levels of p, mo, and sb were below the detection limit, whereas the concentration of other elements such as cr, co, ni, cu, and zn was very high in both surface and deep soil layers (appendix s2 and appendix s3). specifically, in the surface soil layer the concentration of cr varied from a minimum value of 261 to a maximum of 683 ppm at site 1, and from 595 to 1,129 ppm at site 2. in contrast, ni concentration varied from 308 to 554 ppm at site 1, and from 385 to 932 ppm at site 2. the presence of zn was considerable and it also varied from 89 to 114 ppm at site 1, and from 64 to 120 ppm at site 2. similar to the surface layer, we detected a high concentration of cr, ni, and zn in the deep soil layer as well. more precisely, in this soil portion, the content of cr varied from 298 to 915 ppm at site 1, and from 720 to 1,216 ppm at site 2. the ni concentration ranged from 516 to 915 ppm at site 1, and from 413 to 959 ppm at site 2. finally, the content of zn varied from a minimum value of 79 or 96 to a maximum of 115 ppm at sites 1 and 2, respectively. soil mineralogy, lithology, and chemistry of the sites in the soil samples collected at a depth between 5 and 20 cm, we identified the presence of 26 elements (tab. 4), including a high concentration of some microelements (cr, ni, and zr) (appendix s4, s1–s20). more precisely, the highest range of cr (75–1,871 ppm), ni (257–1,371 ppm), and zr (97–305 ppm) values were detected at site 1. the distribution of macroelements was similar at both the sites. the geology of site 1 is different than that of site 2 (tab. 5). site 1 is characterized by a high level of geodiversity: the parent rock consists of siliciclastic conglomerate, serpentine schist, and calceschist; whereas, the site 2 is characterized by only calceschist. the two sites showed variable ph values according to the parent rock (see caption in appendix s4). the lowest ph value corresponds to siliciclastic conglomerate; whereas, the highest values were found on calceschist and serpentine schist. more precisely, site 1 soil samples had ph values ranging from 4.27 to 5.83, whereas at site 2, the ph varied from 4.20 to 4.90. finally, according to grain size analysis (tab. 5), the soil texture was classified as gravelly sand to muddy gravel for site 1, and sandy gravel to gravelly mud for site 2. tab. 1 summary of the element content detected in 20 mushroom samples. macroelements (wt %) microelements (ppm) ca ti mn fe cr ni cu zn sr sb site 1 mean 0.128 0.004 0.007 0.022 40 55 48 81 1 60 sd 0.061 0.000 0.008 0.032 9 60 17 18 1 20 min 0.061 0.004 0.002 0.001 30 5 24 40 1 13 max 0.269 0.004 0.025 0.095 58 209 73 105 3 84 site 2 mean 0.126 0.031 0.006 0.265 45 32 36 152 8 67 sd 0.068 0.040 0.006 0.337 18 24 21 59 11 32 min 0.057 0.004 0.001 0.001 29 6 20 110 1 13 max 0.294 0.104 0.017 0.907 90 92 86 311 28 140 mushroom samples are labeled m1–m20 in appendix s1. 6 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers ta b. 2 su m m ar y of th e el em en t c on te nt d et ec te d in 2 0 so il sa m pl es fr om th e su rf ac e la ye r (0 to 2 0 cm ). m ac ro el em en ts (w t % ) m ic ro el em en ts (p pm ) m g a l si p k c a t i m n fe s c r c o n i c u z n r b sr z r n b m o sn sb b a p b c d v si te 1 m ea n 5. 04 12 .6 4 47 .0 6 0. 11 1. 29 3. 80 0. 89 0. 50 7. 51 6, 59 7 43 8 57 43 3 41 10 0 70 92 15 4 12 2 40 27 21 98 74 18 97 sd 0. 64 1. 32 4. 88 0. 11 0. 32 0. 75 0. 16 0. 25 1. 28 4, 56 9 16 1 19 78 5 9 13 5 35 58 41 10 8 8 16 7 52 m in 4. 06 10 .2 9 38 .5 5 0. 05 1. 00 2. 77 0. 74 0. 26 5. 62 1, 76 3 26 1 18 30 8 34 89 47 82 94 45 2 10 13 83 40 9 23 m ax 5. 78 14 .6 9 55 .2 6 0. 41 1. 83 4. 61 1. 10 0. 84 9. 38 14 ,2 74 68 3 80 55 4 50 11 4 88 10 0 19 6 21 3 95 36 30 11 1 10 3 27 14 1 si te 2 m ea n 11 .9 1 12 .0 9 45 .1 2 0. 09 1. 29 2. 51 1. 09 0. 18 9. 08 2, 64 5 84 5 69 62 4 33 94 83 97 17 4 54 43 37 21 12 4 46 25 17 0 sd 4. 72 1. 82 2. 93 0. 10 0. 46 0. 65 0. 23 0. 03 0. 76 2, 03 5 17 0 16 17 7 4 17 26 10 35 39 53 7 10 52 26 3 54 m in 5. 07 9. 28 39 .2 9 0. 05 0. 59 1. 52 0. 78 0. 13 8. 00 93 5 59 5 39 38 5 25 64 42 83 11 7 2 2 27 2 72 13 21 23 m ax 20 .2 5 15 .5 5 50 .4 6 0. 34 2. 08 3. 98 1. 58 0. 22 10 .5 6 7, 77 8 1, 12 9 10 2 93 2 40 12 0 12 5 11 5 23 8 11 1 10 5 46 32 23 9 85 31 21 1 su rf ac e so il la ye r sa m pl es a re la be le d sl 1– sl 20 in a pp en di x s2 . ta b. 3 su m m ar y of th e el em en t c on te nt d et ec te d in 2 0 so il sa m pl es fr om th e de ep la ye r (2 0– 40 c m fr om th e to p of th e so il) . m ac ro el em en ts (w t % ) m ic ro el em en ts (p pm ) m g a l si p k c a t i m n fe s c r c o n i c u z n r b sr z r n b m o sn sb b a p b c d v si te 1 m ea n 7. 65 14 .3 2 49 .8 0 0. 05 1. 08 2. 42 1. 08 0. 17 9. 84 1, 40 7 60 5 64 70 2 42 86 58 81 18 6 53 40 41 21 12 1 52 32 13 8 sd 1. 04 0. 52 2. 20 0. 00 0. 16 0. 30 0. 09 0. 03 0. 65 38 6 27 8 8 17 6 6 5 12 11 16 7 21 5 7 28 14 3 19 m in 6. 78 13 .3 0 45 .3 1 0. 05 0. 79 2. 08 0. 98 0. 13 8. 93 82 7 29 8 54 51 6 35 79 40 60 14 9 35 2 32 13 79 31 28 10 4 m ax 9. 85 15 .0 8 52 .5 6 0. 05 1. 35 3. 01 1. 28 0. 21 11 .1 1 2, 02 3 91 5 80 91 5 55 96 67 93 20 2 60 60 50 29 17 0 82 37 17 0 si te 2 m ea n 11 .7 5 13 .1 9 46 .5 1 0. 08 1. 33 2. 04 0. 99 0. 15 10 .0 3 1, 93 0 94 0 75 66 3 33 94 72 93 14 7 48 42 38 20 15 2 53 28 16 2 sd 5. 18 1. 45 1. 73 0. 03 0. 56 0. 55 0. 14 0. 04 0. 57 1, 00 2 15 8 28 16 8 6 12 30 10 29 22 23 7 9 39 27 5 58 m in 6. 01 11 .2 5 43 .3 2 0. 05 0. 05 1. 41 0. 75 0. 10 8. 92 1, 12 9 72 0 3 41 3 23 79 25 78 10 1 0 2 22 13 89 18 17 23 m ax 20 .4 4 15 .8 2 49 .9 5 0. 11 2. 26 3. 29 1. 20 0. 22 11 .0 8 4, 21 0 1, 21 6 10 9 95 9 44 11 5 13 6 10 7 21 2 85 70 51 30 23 9 11 1 35 24 3 d ee p so il sa m pl es a re la be le d d l1 –d l2 0 in a pp en di x s3 . 7 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers ta b. 5 ph ys ic al a nd g eo lo gi ca l s oi l f ea tu re s of e ac h si te . ph g ra nu lo m et ry r oc k ty pe % c la y (< 2 m m ) % s ilt (2 m m –0 .6 3 µm ) % s an d (< 63 µ m ) si te 1 m ea n 5. 23 23 .1 4 26 .6 7 50 .1 9 se rp en tin es ch is ts c al ce sc hi st s c hl or ite -a ct in ol ite s ch is ts c on gl om er at es sd 0. 52 6. 90 5. 86 10 .4 6 m in 4. 27 15 .1 3 16 .3 1 29 .9 4 m ax 5. 83 35 .6 4 35 .0 7 68 .5 6 si te 2 m ea n 4. 62 50 .0 8 32 .2 8 17 .6 5 c al ce sc hi st s sd 0. 22 15 .9 9 10 .1 9 14 .0 3 m in 4. 20 23 .3 4 19 .4 9 4. 88 m ax 4. 90 75 .6 2 45 .3 2 55 .3 4 ta b. 4 su m m ar y of th e el em en t c on te nt d et ec te d in 2 0 so il sa m pl es c ol le ct ed a t d ep th s be tw ee n 5 an d 20 c m fr om th e to p of th e so il. m ac ro el em en ts (w t % ) m ic ro el em en ts (p pm ) si t i a l fe m n m g c a k s v c r c o n i c u z n r b sr z r n b m o c d sn sb b a p b si te 1 m ea n 48 .5 7 1. 75 15 .6 1 10 .9 1 0. 17 7. 73 2. 53 1. 01 1, 20 0 16 0 81 9 94 67 8 53 78 48 10 3 19 0 35 16 31 44 18 16 7 40 sd 4. 46 0. 53 2. 09 2. 42 0. 09 2. 96 0. 96 0. 29 19 0 39 62 7 49 30 3 23 10 15 35 62 20 20 4 7 16 26 9 m in 39 .8 3 1. 02 12 .6 9 8. 49 0. 10 3. 09 1. 90 0. 70 1, 00 9 99 75 45 25 7 28 65 17 64 97 0 0 26 31 0 11 5 17 m ax 54 .5 7 2. 86 19 .8 7 15 .8 3 0. 40 13 .2 0 5. 15 1. 62 1, 59 1 22 3 1, 87 1 21 9 1, 37 1 10 3 93 69 18 6 30 5 57 40 39 53 35 21 5 49 si te 2 m ea n 49 .1 1 1. 37 16 .6 2 10 .4 0 0. 17 5. 82 1. 47 2. 00 87 2, 06 1 18 0 21 4 59 15 2 32 91 12 0 11 7 20 6 84 45 29 41 15 4 63 sd 4. 17 0. 25 2. 09 2. 02 0. 14 2. 07 0. 43 0. 71 3 94 2 24 57 19 85 14 25 42 20 47 37 40 6 12 38 19 m in 42 .6 8 1. 09 11 .9 3 6. 94 0. 06 4. 45 1. 03 0. 82 83 1, 01 2 14 6 14 6 28 30 13 41 62 90 13 7 51 0 17 27 10 3 38 m ax 58 .4 7 1. 84 18 .9 4 12 .7 8 0. 43 11 .4 3 2. 15 3. 19 94 4, 50 2 21 3 32 3 86 30 9 59 11 7 20 2 15 5 27 5 17 5 13 1 37 58 23 9 97 th es e sa m pl es a re la be le d s1 –s 20 in a pp en di x s4 . 8 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers chemical correlation among mushrooms and soil layers in order to observe the differences between the element concentration detected in sporocarps and the underlying soil layers, we used a graph based on descriptive statistical measurements (see “material and methods”). fig. 3 and fig. 4 display the results of the macroand microelement content detected in mushrooms and soil samples from sites 1 and 2, respectively. in both sites, the mushroom content differed from the soil samples for some macroelements (fe, ca) (fig. 3). regarding microelement content, the concentration of zn, cu, sr, and sb did not differ between the mushrooms and the soil layers (fig. 4). conversely, the content of cr and ni was lower in the mushrooms than in the soil portions, where we found a high concentration of these elements. the degree of correlation among the element contents detected in each sample level was also calculated by pearson’s coefficient. overall, the highest correlation values were observed among the soil layers for the majority of the detected elements at both sites. for example, a strong positive correlation value (r = 0.91) was obtained for cr and zn detected in the soil layers in both sites 1 and 2, respectively (tab. 6). the element content measured in the mushrooms, however, did not show strong positive correlation values compared to those detected in the underground soil samples. the highest positive correlation value in this group (r = 0.57) was observed between the mn content quantified in the mushrooms and in the surface soil layer (tab. 6). ns ns ns ns 0.0 0.5 1.0 1.5 2.0 cao tio2 mnofe2o3 chemical element pp m a *** * ** ** 0.0 2.5 5.0 7.5 10.0 12.5 cao tio2 mnofe2o3 chemical element b ns ns ns ns 0.0 2.5 5.0 7.5 10.0 12.5 cao tio2 mnofe2o3 chemical element c fig. 3 boxplot of macroelement content in mushrooms (a), surface soil layer (b), and deep soil layer (c) collected at site 1 (red) and site 2 (blue). the element concentration is expressed in ppm. statistical significance: **** α = 0.001; *** α = 0.01; ** α = 0.05; * α = 0.1; ns – not significant values. ns ns ns **** ns ns 0 50 100 150 200 cr ni cu zn sr sb chemical element pp m a **** ** *** ns ns ns 0 400 800 1,200 cr ni cu zn sr sb chemical element b ** ns ** ns * ns 0 400 800 1,200 cr ni cu zn sr sb chemical element c fig. 4 boxplot of microelement content in mushrooms (a), surface soil layer (b), and deep soil layer (c) collected at site 1 (red) and site 2 (blue). element concentration is expressed in ppm. statistical significance: **** α = 0.001; *** α = 0.01; ** α = 0.05; * α = 0.1; ns – not significant values. 9 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers multivariate analyses a set of multivariate analyses was used to measure the degree of dissimilarity among the mushroom and soil layer samples. in detail, the result of cluster analysis (fig. 5, fig. 6) showed that the mushrooms and soil samples do not form clearly distinctive clusters based on origin site with regard to macroelements (fig. 5). instead, microelement concentrations define separate groups based on geographic site of growth (fig. 6). the pca analysis showed that two axes explained 76.1% of the total data variance (fig. 7). specifically, when considering all the samples together, the different concentration of some macro(ca, fe, ti) and microelements (sb, zn, cu, cr, ni) separate the samples by spatial distribution (m vs. dl and sl). the isa results (tab. 7) emphasize that some elements had a significant indicator value (iv). specifically, we found that some macroelements, such as fe, sr, and mn, were significant for mushroom samples, whereas ca and ti were significant elements for the deep soil layer. for both the surface and deep soil layers, cr was a common significant element. discussion our results supported the hypothesis that the soil lithology, mineralogy, and chemistry of a site can influence the element content in porcini (here boletus reticulatus). we showed that at both study sites, most elements detected in the sporocarps almost completely reflected the content detected in the underlying soil layers (fig. 3, fig. 4). in more detail, we showed that the concentration of some microelements (cu, zn, sr, sb) detected in the mushrooms is very similar to that measured in both the soil layers (fig. 4). conversely, some other elements (cr and ni) showed a different distribution. the distribution of macroelements appeared to be variable among the mushrooms and the underlying soil samples, especially that of ca and fe. we surmised that the variation in the macroelement content is due to the wide distribution that these elements have in soil. the correlation values obtained (tab. 5) also emphasized the above-described element distribution. the highest correlation values were observed between the soil portions, sl and dl (tab. 5), rather than the same elements compared between either soil layer and the mushroom samples. moreover, the strongest pearson’s values (0.61 < r < 0.91) were obtained by performing the correlation between the element content in the surface layer and deep soil, confirming that some microelements have low mobility between soil layers. tab. 6 pearson correlation values (r) among the mushroom samples (m) versus the soil layers (surface soil – sl and deep soil – dl). samples code elements ca ti mn fe cr ni cu zn sr sb total m vs. sl 0.10 0.14 0.40 0.23 0.28 −0.34 −0.03 −0.47 −0.24 0.16 m vs. dl 0.06 −0.55 −0.33 −0.30 0.04 −0.15 0.00 0.07 −0.09 0.36 sl vs. dl 0.48 0.29 0.34 0.29 0.87 0.11 0.73 0.65 0.45 −0.19 site 1 m vs. sl 0.35 0.00 0.57 −0.58 0.03 −0.30 −0.68 0.03 0.12 0.04 m vs. dl −0.21 0.00 −0.25 −0.47 −0.13 −0.10 −0.28 −0.28 −0.30 0.56 sl vs. dl −0.44 0.79 0.34 0.04 0.91 −0.71 0.68 0.69 −0.24 −0.04 site 2 m vs. sl −0.11 −0.11 −0.44 −0.08 0.37 −0.39 −0.08 −0.52 −0.51 0.22 m vs. dl 0.18 −0.55 −0.52 −0.60 −0.08 −0.49 −0.24 −0.32 −0.64 0.29 sl vs. dl 0.79 0.50 0.78 0.64 0.68 0.68 0.33 0.91 0.71 −0.28 positive moderate (0.40 < r < 0.59), strong (0.60 < r < 0.79), and very strong (0.80 < r < 1) correlation values are indicated in bold. 10 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers m 1 m 5 m 3 m 4 m 20 m 7 m 13 m 2 m 14 m 8 m 9 m 6 m 10 m 15 m 19 m 11 m 18 m 12 m 16 m 17 sl 1 sl 11 sl 3 sl 4 sl 5 sl 14 sl 20 dl 11 sl 2 sl 16 sl 17 dl 4 dl 6 dl 10 sl 15 sl 19 sl 13 dl 1 dl 2 dl 5 dl 8 dl 9 dl 16 dl 14 dl 17 dl 15 dl 18 dl 20 sl 18 dl 3 dl 19 dl 7 dl 12 sl 12 dl 13 sl 6 sl 8 sl 9 sl 10 sl 7 0. 0 0. 2 0. 4 0. 6 0. 8 1. 0 he ig ht site 1 site 2 m 1 m 2 m 10 m 3 m 6 m 5 m 7 m 8 m 9 m 11 m 16 m 17 m 12 m 13 m 20 m 19 m 15 m 14 m 4 m 18 sl 1 sl 3 sl 2 sl 4 dl 4 dl 1 dl 5 dl 3 dl 2 sl 5 sl 6 dl 13 dl 14 dl 18 sl 8 sl 13 sl 9 sl 14 sl 20 s l7 sl 10 sl 11 sl 19 sl 18 dl 16 dl 19 sl 15 dl 20 dl 15 dl 17 sl 16 sl 17 dl 6 dl 8 dl 10 dl 7 dl 9 dl 11 sl 12 dl 12 0. 0 0. 2 0. 4 0. 6 he ig ht site 1 site 2 −3 0 3 6 −5.0 −2.5 0.0 2.5 dim1 (63.7%) d im 2 (1 2. 4% ) sample dl m sl a cao tio2 mno fe2o3 cr ni cu zn srsb −1.0 −0.5 0.0 0.5 1.0 −1.0 −0.5 0.0 0.5 1.0 dim1 (63.7%) d im 2 (1 2. 4% ) 0.25 0.50 0.75 cos2 b fig. 5 cluster dendrogram of the samples (mushrooms, surface soil layer, and deep soil) based on their macroelement content. designations of m1–20, sl1–20, and dl1–20 indicate the mushrooms (m), surface soil layer (sl), and deep soil samples (dl), respectively. fig. 6 cluster dendrogram of the samples (mushrooms, surface soil layer, and deep soil) based on microelement content. designations of m1–20, sl1–20, and dl1–20 indicate the mushrooms (m), surface soil layer (sl), and deep soil samples (dl), respectively. fig. 7 graphical representation of principal component analysis. (a) ordination graph of the first two principal components. ellipses are drawn around the 95% confidence interval for each sample group centroid. (b) correlation circle between a variable and a principal component (pc). the cos2 value indicates the quality of representation of the variable on the principal component. m – mushroom samples; sl – surface soil samples; dl – deep soil samples. 11 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers the degree of chemical dissimilarity among the samples is also shown by the cluster analysis (ca) result (fig. 5, fig. 6). both dendrograms indicate that mushroom samples form a distinctive group apart from the soil layers. this confirms that our collected mushrooms had a different chemical content for some elements (ca, fe, ni, and cr), than that of the soil layers. the analyzed mushrooms have accumulated, in fact, a minority of elements (10 of the 26 analyzed) from the soil. moreover, the ca results also displayed a high degree of similarity between the surface and deep soil samples, establishing a unique group in the cluster dendrograms. the result of ca also showed that considering the microelement concentration, mushroom samples (m1– m10 and m11–m20) as well as soil samples established distinctive groups on the basis of their geographical sites of origin (fig. 6). this result supported our hypothesis that soil geology of origin influences the chemical composition of wild edible mushrooms, and suggested that some microelements can be used as fingerprints to indicate the geographic provenance of a sample. additionally, the pca result confirmed that when considering the whole chemical element pattern, the mushrooms samples formed a distinctive group (fig. 7a) because of their higher content of sb, zn, and cu than the soil layers (fig. 7b). the chemical analysis performed on our mushrooms and soil samples also revealed the presence of some toxic elements in the two sites: zn, ni, cr, co, and cu. however, it is important to highlight that the high concentrations of some heavy metals (e.g., cr) in the two ligurian sites is due to natural geological background factors (in this case, the presence of serpentine schist parent rocks) of this geographic area, rather than an anthropogenic source of pollution. the concentration of these elements is in fact very common, and high in soil developed on ophiolitic and ultrabasic rocks [57–59]. the presence of toxic elements in mushrooms confirms the mushrooms’ ability to take up heavy metals from the growing substratum [1–4,7]. however, it should also be emphasized that the content of cr detected in the sporocarps samples did not exceeded the limits of the law, in which tolerable intake values for heavy metals are set by regulatory agencies [14,15,60]. comparing our results with those from different studies is quite difficult, since very few studies fully describe the chemical content of wild edible mushrooms, especially porcini, as well as the geology of their sites of growth/origin. based on the available literature, interesting aspects emerge from the comparison of our results with those obtained by nonnis marzano et al. [10]. these authors analyzed trace element concentration in some boletus species recorded in central italy. despite a different geology in their studied area, concentrations of zn in both the mushrooms and the soil samples were very similar to our results. also, giannaccini et al. [61] analyzed the content of microelements in the (top)soil and in some edible mushrooms (viz. b. edulis and macrolepiota procera), growing on sedimentary-clastic rocks (prevalently limestone), in central italy (tuscany). specifically, this group found a level of zn similar to our results in both b. edulis and soil samples. based on these similarities, it may be supposed that some specific italian sites favorable for the growth of porcini are characterized by similar levels of zn ([zn] > 100 ppm ca. in the mushrooms and [zn] < 100 ppm ca. in the soil), despite differing in soil geology (for parent rock). in contrast, as variables influenced by natural geological background factors, the cr, ni, and sb content detected in our sites (and mushroom samples) was higher compared to the other italian areas [7,10,61]. moreover, in another recent study on the element content of boletus aereus from volcanic areas in south italy (sicily), the authors found a high concentration of z, zn, cu, se, and ti in the analyzed fungal samples [62]. despite some evidence that emerged from comparing our results with those of nonnis marzano et al. [10], characterization of the soil chemistry of naturally-growing porcini habitats is quite difficult to perform. based on these gaps in knowledge, we performed statistical analysis (tab. 7) to detect possible soil and mushroom chemical tab. 7 summary of the isa technique. element sample iv p ca dl 0.746 0.03* ti dl 0.730 0.03* fe m 0.997 0.010** sr m 0.926 0.020* mn m 0.890 0.040* cr dl 0.853 0.005** cr sl 0.848 0.005** ni dl 0.759 0.015* m, sl, and dl indicate mushrooms, surface soil layer, and deep soil samples, respectively. iv – indicator value. significance: *** p = 0; ** p = 0.001; * p = 0.01. 12 of 15© the author(s) 2019 published by polish botanical society acta mycol 54(2):1130 ambrosio et al. / chemical elements in mushrooms and soil layers indicators. based on our results, some elements (ca, ti, mn, sr, and cr), had a significant value, which indicated that they may be considered potential soil and mushroom geomarkers. in conclusion, the results obtained by our study highlight that mushrooms and soil samples, with different substrata of origin, differ considerably in their chemical components, especially with regard to microelements. this finding leads us to recommend that geological and chemical soil information should be included in food traceability. based on our results, the application of this method could be the basis for performing quality assurance on natural products. the study of chemical content in the mushrooms and in the soil layers can protect genuine products and distinguish them from uncertified and unknown-origin products. since the application of geochemical approaches in the mycological field has not been widely adopted, future efforts should include more extensive sampling to implement this method in protecting human health and food safety. acknowledgments authors are grateful to g. nardi for the graphic images, to f. boccardo for his collaboration in the field survey and the picture of porcini, and to the managing/production editor and t. hunt for linguistic revision. anonymous reviewers are also acknowledged for useful comments and observations. supplementary material the following supplementary material for this article is available at http://pbsociety.org.pl/ journals/index.php/am/rt/suppfiles/am.1130/0: appendix s1 element content detected in the mushroom samples. appendix s2 element content in surface soil layer (0–20 cm) samples (sl1–sl20). appendix s3 element content in deep soil layer (20–40 cm) samples (dl1–dl20). appendix s4 element content in soil layer (5–20 cm) samples (s1–s20). references 1. demirbas a. accumulation of heavy metals in some edible mushrooms from turkey. food chem. 2000;68:415–419. https://doi.org/10.1016/s0308-8146(99)00210-1 2. svoboda l, havlíčková b, kalač p. contents of cadmium, mercury and lead in edible mushrooms growing in a historical silver-mining area. food chem. 2006;96:580–585. https://doi.org/10.1016/j.foodchem.2005.03.012 3. yamac m, yildiz d, sarikurkcu c, celikkollu m, solak mh. heavy metals in some edible mushrooms from the central anatolia, turkey. food chem. 2007;103(2):263–267. https://doi.org/10.1016/j.foodchem.2006.07.041 4. zhang d, gao t, ma p, luo y, su p. bioaccumulation of heavy metal in wild growing mushrooms from liangshan yi nationality autonomous prefecture, china. wuhan university journal of natural sciences. 2008;13(3):267–272. https://doi.org/10.1007/s11859-008-0302-2 5. kirk pm, cannon pf, minter dw, stalpers ja. dictionary of the fungi. 10th ed. wallingford: cab international; 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interpopulation variability; randomly amplified polymorphic dna; rapd; hypogymnia physodes; sand dunes introduction hypogymnia physodes (l.) nyl. is an epiphytic species of lichenized fungi of the family parmeliaceae. this taxon also occurs, although less often, on bedrock, mosses, or soil. hypogymnia physodes produces numerous soredia and reproduces mainly vegetatively. recent molecular studies have shown that genetic diversity in the lichens reproducing vegetatively is high and not lower than in the lichens that reproduce sexually [1–3]. hypothetically, the preference of lichens for one type of substrate can be reflected in their genetic constitution. using molecular markers, mattsson et al. [4] have distinguished among populations of h. physodes, growing on different types of trees, the haplotypes that more than others prefer a specific type of substrate. the primary objective of this study was to evaluate the interpopulation variability of epigeic populations of h. physodes growing on the sand dunes of the polish seacoast and their preference for substrate. the genetic variability of h. physodes was studied using randomly amplified polymorphic dna (rapd) analyses, although rapd analyses have been still rarely used in lichenized fungi [5–7], possibly due to their low reproducibility. doi: 10.5586/am.1096 publication history received: 2016-04-27 accepted: 2017-05-01 published: 2017-07-17 this article was originally submitted to and processed by the acta agrobotanica (another journal of the polish botanical society) editors. upon consent of the authors and the editors of both journals, it was eventually published in acta mycologica. handling editor joanna kaczmarek (acta agrobotanica), institute of plant genetics, polish academy of sciences, poland authors’ contributions aw: idea of the study, writing the manuscript, collection of material; ma: writing the manuscript, final version of the manuscript, genetic analyzes; at: genetic analyzes, manuscript drafting; ał: collection of material, preparation of the distribution map; ap: collection of material, final version of the manuscript funding the research has been conducted on the authors own expenses. competing interests no competing interests have been declared. copyright notice © the author(s) 2017. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation wieczorek a, achrem m, truszkowska a, łysko a, popiela a. genetic diversity of natural psammophilous populations of hypogymnia physodes (l.) nyl. on polish seacoast dunes. acta mycol. 2017;52(1):1096. https:// doi.org/10.5586/am.1096 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:anetta.wieczorek%40usz.edu.pl?subject=genetic%20diversity%20of%20natural%20psammophilous%20populations%20of%20hypogymnia%20physodes%20%28l.%29%20nyl.%20on%20polish%20seacoast%20dunes https://doi.org/10.5586/am.1096 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ https://doi.org/10.5586/am.1096 https://doi.org/10.5586/am.1096 2 of 6© the author(s) 2017 published by polish botanical society acta mycol 52(1):1096 wieczorek et al. / genetic diversity of hypogymnia physodes based on rapd material and methods lichen material samples of hypogymnia physodes were collected on the sand dunes of the polish seacoast (fig. 1) from 11 sites in july 2014. the analysis was performed on dried and cleaned material. dna extraction and rapd analysis genomic dna from each population was isolated using the dneasy plant mini kit (qiagen, usa). rapd analysis was then carried out according to the modified method of willliams et al. [8]. amplification reactions were carried out in 25 µl final volume of reaction mixture containing: 50 ng dna, 1× green gotaq reaction buffer (promega, usa), 0.2 mm dntps (fermentas, lithuania), 0.28 mm mgcl2, 5 mm primers, 0.04 u gotaq dna polymerase (promega). the thermal profile of the reaction was determined experimentally and included initial dna denaturation at 95°c for 11 min, 40 cycles of dna denaturation at 93°c for 30 s, primer annealing at 37°c for 40 s, dna chain elongation at 72°c for 80 s, and final elongation at 72°c for 5 min. amplification was carried out twice in an mj mini gradient thermal cycler (bio-rad, usa) on two samples of dna from each genotype. the resulting amplification products were separated on 2.0% agarose gel with ethidium bromide (5 µg/ml; sigma-aldrich, usa). data analysis to visualize, document, and analyze the results obtained, a set of gel doc xr and quantity one 4.6.5 software (bio-rad) were used. in the populations, we assumed the existence of hardy–weinberg equilibrium. the genetic similarity of the studied populations of hypogymnia physodes was also determined using similarity index, according to dice’s formula (1945), as described by nei and li [9]. the genetic similarity matrix was used to construct the upgma dendrogram using the freetree and treeview 1.6.6 software [10,11]. fig. 1 study area and location of sampled populations 1–11 of hypogymnia physodes. 3 of 6© the author(s) 2017 published by polish botanical society acta mycol 52(1):1096 wieczorek et al. / genetic diversity of hypogymnia physodes based on rapd results this study examined the variation of 11 populations represented by 45 specimens on the basis of 133 loci, i.e., rapd bands, in which a model of total dominance was assumed. no band was defined as a recessive allele and, on this ground, the probable percentage of recessive alleles in the population was calculated on the basis of bayesian method that estimates the allele frequency based on the diversity of rapd fragments, separately in all the analyzed loci in each population. the total number of analyzed pcr products was 153 and included polymorphic products whose size ranged from 155 to 1500 base pairs (tab. 1, fig. 2). the number of the polymorphic products amplified by one (individual) primer ranged from 0 in lich2 to 55 in c02. the assay effectiveness index (effective multiplex ratio; emr) was also determined, as a ratio of the number of polymorphic products to the number of applied primers, which amounted to 14.78. the highest similarity values were recorded for population from gąski: 73% similarity to the population from łeba and 70% similarity to the population from pobierowo (tab. 2). the analyses showed the lowest genetic similarity for populations from wisełka and międzywodzie versus the populations from pobierowo and darłówko (tab. 2). overall, the lowest genetic similarity (0–45%) was characteristic for the hypogymnia physodes population from międzywodzie. dendrogram (fig. 3) revealed two separate clusters: the first group is formed by the lichen populations from międzyzdroje, międzywodzie, and wisełka, found on the bark of pine trees, whereas the second one includes the other analyzed populations of h. physodes, occurring on sand. this is confirmed by clade stability tests, with the use of bootstrap resampling methods. discussion the study of molnár and farkas [12], in which the five loci sequences were analyzed, found no significant genetic diversity of the h. physoides population. this is also confirmed by studies by mattsson et al. [4]. tab. 1 primers employed with their sequence, the number of rapd markers obtained, as well as the number and percentage (p) of polymorphic markers for each primer. primer name sequence 5'–3' total no. of products no. of polymorphic products p (%) c02 gtgaggcgtc 55 55 100 d03 gtcgccgtca 23 23 100 d08 gtgtgcccca 24 23 96 e07 agatgcagcc 12 9 75 lich2 aacgcgcaac 6 0 0 lich3 gtagacccgt 8 3 37 lich4 aagagcccgt 13 12 92 lich5 cccgtcagca 10 6 60 opa 18 aggtgaccgt 2 2 100 total 153 133 fig. 2 agarose gel electrophoresis of pcr product from dna od hypogymnia physodes obtained with primer c02. 4 of 6© the author(s) 2017 published by polish botanical society acta mycol 52(1):1096 wieczorek et al. / genetic diversity of hypogymnia physodes based on rapd however, our analyzes showed a high population differentiation depending on the presence on the bark or on the dunes. as a result of the reactions being performed, 87% of the products were polymorphic, which indicates a high genetic diversity of the analyzed populations of hypogymnia physodes. among the lichen populations growing on tree bark, forming the first group, the highest genetic similarity was observed between individuals from międzywodzie and międzyzdroje (64%); a lower genetic similarity was found between other populations, not exceeding 50% (tab. 2). such a high genetic diversity between local populations of lichens has been already described [2,13]. moreover, another studies have shown even higher intraspecific genetic diversity than noted in our research, e.g., in the lichen species lobothallia alphoplacaand, l. radiosa [14] or in the fungus pisolithus tinctorius [15]. the reason of such high genetic may be the habitat whose isolation reduces the flow of genes between populations [16–18]. the high variation is an important trait because it plays a role under environmental changes, determining the survival of organisms [14,19,20]. the evaluation of two analyzed groups of h. physodes, the first growing on the bark of pine trees and the second on the sand, confirms the hypothesis that lichens are sensitive to substrate features [21]. it is believed that some genotypes are closely related to a preferential substrate, which can be associated with the process of colony formation [22]. the fact that the type of substrate can affect lichen variation is confirmed by other studies of h. physodes, in which an intraspecific diversity resulting from different amounts of nutrients in the substrate has been shown [4]. a high diversity, as in h. physoides, between populations preferring different substrate was also observed in xanthoria parietina [17]. sequence analysis of the intergenic spacer (igs) and internal transcribed spacer (its) sequences revealed significant differentiation between seven populations tab. 2 dice’s similarity matrix of hypogymnia physodes populations. population 1miz 2wis 3miw 4pob 5pus 6sia 7gas 8dar 9ust 10leb 2wis 0.48 3miw 0.64 0.43 4pob 0.28 0.00 0.00 5pus 0.58 0.32 0.39 0.28 6sia 0.50 0.20 0.36 0.62 0.35 7gas 0.63 0.46 0.45 0.48 0.70 0.55 8dar 0.41 0.06 0.08 0.66 0.53 0.54 0.51 9ust 0.44 0.23 0.30 0.40 0.60 0.58 0.54 0.46 10leb 0.58 0.35 0.32 0.48 0.62 0.62 0.73 0.64 0.66 11lub 0.56 0.33 0.36 0.62 0.41 0.40 0.68 0.45 0.25 0.48 10leb 7gos 5pus 9ust 4pob 8dar 6sia 11lub 1miz 3miw 2wis 34 18 24 9 18 41 24 56 23 100 0,1 g r o u p i g r o u p i i fig. 3 upgma dendrogram of hypogymnia physodes populations based on rapd markers. 5 of 6© the author(s) 2017 published by polish botanical society acta mycol 52(1):1096 wieczorek et al. / genetic diversity of hypogymnia physodes based on rapd xanthoria parietina in different habitat (on the bark and rock) but, in contrast to our results, it was not high between the studied populations within the same habitat type [17]. results reported by gaya et al. 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https://doi.org/10.1890/0012-9658(2006)87[2037:qdaeli]2.0.co;2 https://doi.org/10.1073/pnas.1507072112 abstract introduction material and methods lichen material dna extraction and rapd analysis data analysis results discussion references 2017-07-17t17:50:13+0100 piotr otręba preliminary studies of fungi in the biebrza national park. part iv. macromycetes – new data and the synthesis 1 of 28published by polish botanical society acta mycologica original research paper preliminary studies of fungi in the biebrza national park. part iv. macromycetes – new data and the synthesis anna kujawa1*, błażej gierczyk2, grażyna domian3, marta wrzosek4, małgorzata stasińska5, jarosław szkodzik6, tomasz leski7, leszek karliński7, marcin pietras7,8, maria dynowska9, agnieszka henel10, dominika ślusarczyk11, dariusz kubiak9 1 institute for agricultural and forest environment, polish academy of sciences, bukowska 19, 60-809 poznań, poland 2 faculty of chemistry, adam mickiewicz university in poznań, umultowska 89b, 61-614 poznań, poland 3 regional directorate for environmental protection in szczecin, teofila firlika 20, 71-637 szczecin, poland 4 department of molecular phylogenetics and evolution, university of warsaw, al. ujazdowskie 4, 00-478 warsaw, poland 5 environmental testing laboratory, university of szczecin, felczaka 3c, 71-412 szczecin, poland 6 nature & ecology of łódź macroregion website, ekolodzkie.pl 7 institute of dendrology, polish academy of sciences, parkowa 5, 62-035 kórnik, poland 8 faculty of biology, university of gdańsk, wita stwosza 59, 80-308 gdańsk, poland 9 department of mycology, university of warmia and mazury in olsztyn, oczapowskiego 1a, 10-957 olsztyn, poland 10 biebrza national park, osowiec-twierdza 8, 19-110 goniądz, poland 11 department of algology and mycology, faculty of biology and environmental protection, university of łódź, banacha 12/16, 90-237 łódź, poland * corresponding author. email: annakuja@poczta.onet.pl abstract the paper presents the last part of the results of the short-term inventory of fungi species in the biebrza national park and synthesises all the data gathered during two surveys, including the information published by other authors. the main body of research is focused on a survey of macrofungi identified with morphologybased methods. in case of some specimens molecular technics have been applied. in total 346 macrofungal taxa (21 belonging to ascomycetes and 325 to basidiomycetes) were found during the survey, including 186 species unobserved during the previous inventory in 2012. as a result of previous and ongoing studies, the current number of macrofungi recorded from the biebrza national park reached 508 species. among them eight taxa are newly reported for poland (conocybe velutipes var. nitrophila, entoloma caeruleum, e. plebejoides, inocybe rennyi, i. vulpinella, pholiota pityrodes, pholiotina utricystidiata, and tomentella pilosa). the next seven species (bovista paludosa, fistulina hepatica, ganoderma lucidum, geastrum schmidelii, inonotus obliquus, tulostoma kotlabae, and xerocomus parasiticus) are protected by law and 95 species belong to red-listed species. the results of two intensive, but relatively short-term survey clearly indicate the biebrza national park as a hot spot of macrofungi and suggest the need to undertake extended and regular inventories also in other polish national parks. keywords macrofungi; rare species; protected and threatened fungi; protected area; poland doi: 10.5586/am.1070 publication history received: 2016-01-12 accepted: 2016-01-21 published: 2016-01-29 handling editor tomasz leski, institute of dendrology of the polish academy of sciences, poland authors’ contributions all authors contributed to the collection of material and manuscript preparation. funding the study was privately funded. additional funding was provided by the institute for agricultural and forest environment of the polish academy of science and institute of dendrology of the polish academy of science. competing interests tl is the editorial secretary of the acta mycologica; other authors: no competing interests copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation kujawa a, gierczyk b, domian g, wrzosek m, stasińska m, szkodzik j, et al. preliminary studies of fungi in the biebrza national park. part iv. macromycetes – new data and the synthesis. acta mycol. 2015;50(2):1070. http://dx.doi. org/10.5586/am.1070 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:annakuja%40poczta.onet.pl?subject=preliminary%20studies%20of%20fungi%20in%20the%20biebrza%20national%20park.%20part%20iv.%20macromycetes%20%e2%80%93%20new%20data%20and%20the%20synthesis http://dx.doi.org/10.5586/am.1070 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1070 http://dx.doi.org/10.5586/am.1070 2 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data this paper is dedicated to professor maria lisiewska and professor anna bujakiewicz on the occasion of their 80th and 75th birthday, respectively. introduction the presented data originate from the second survey of macrofungal diversity of the biebrza national park (bbnp) undertaken by members of the polish mycological society (pms) in 2013. that research was a continuation of the all-fungi inventory started in 2012 and summarized by kujawa et al. [1] and ruszkiewicz-michalska et al. [2]. the aim of the current paper is to present the new data supplementing the knowledge of taxonomic diversity and richness of the macrofungi collected also in habitats unscreened during the first survey in 2012 and summarize all known information about macrofungi of the bbnp. the overview of natural values of the bbnp as well as the map indicating new localities surveyed in 2013 was given in the paper concerning new data on micromycetes [3]. material and methods macromycetes were collected 24–29 august 2013 in seven protective units of the bbnp: brzeziny, grzędy, kapice, osowiec, werykle (inventoried previously [1]) and tajno, trzyrzeczki (studied for first time) and at the area of fortress in osowiec-twierdza (see fig. 1 in [3]). the habitats screened include: meadows, pastures, roadsides, bogs and forest communities, including ribeso nigri-alnetum, thelypteridi-betuletum pubescentis, tilio cordatae-carpinetum betuli, and vaccinio uliginosi-pinetum (syntaxonomic system according to matuszkiewicz [4]). more details on the materials and methods used in the paper were presented by kujawa et al. [1]. as in the former paper, the nomenclature follows the checklists [5,6] and the index fungorum database [7]. for some fungi specimens morphological identification has been confirmed using the dna barcoding technique. nuclear its sequences obtained in this study are deposited in national center for biotechnology information (ncbi), with the accession numbers ku525690–ku525701. the voucher collections are deposited in the herbarium of the faculty of biology, university of warsaw, in the institute of dendrology of the polish academy of sciences in kórnik. results the five-days inventory yielded 346 species, mainly basidiomycetes (330 taxa), including 186 species unobserved during the previous inventory in 2012. the following abbreviations are used in the list of species: bpu – brzeziny protective unit (see fig. 1 in ruszkiewicz-michalska et al. [3]); gpu – grzędy protective unit; kpu – kapice protective unit; kopu – kopytkowo protective unit; opu – osowiec protective unit; tpu – trzyrzeczki protective unit; tapu – tajno protective unit; wpu – werykle protective unit; r-a – ribeso nigri-alnetum; t-b – thelypteridi-betuletum pubescentis; t-c – tilio cordatae-carpinetum betuli; v-p – vaccinio uliginosipinetum. initials of the names are given to denote collecting and/or identifying person (i.e., af-s – anna frymark-szymkowiak, ah – agnieszka henel, ak – anna kujawa, bg – błażej gierczyk, ct – cezary tkaczuk, dk – dariusz kubiak, dś – dominika ślusarczyk, gd – grażyna domian, jc – jerzy chełkowski, jp – julia pawłowska, js – jarosław szkodzik, lk – leszek karliński, md – maria dynowska, mp – marcin pietras, mrm – małgorzata ruszkiewicz-michalska, mw – marta wrzosek, ms – małgorzata stasińska, nm – natalia michalska, tl – tomasz leski); – taxon new for bbpn; – taxon new for poland; # – species very rare in poland (1–3 localities); ex, e, v, r, i – category of threat (according to wojewoda, ławrynowicz [8]); pp – partially protected species; sp – strictly protected species. 3 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data list of species collected in 2013 ascomycota ascocoryne cylichnium (tul.) korf on wood, opu, carska droga, mixed forest, 29 aug., leg. & det. mw. chlorociboria aeruginascens (nyl.) kanouse ex c.s. ramamurthi, korf & l.r. batra on wood debris, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. mrm & nm, det. ak; opu, carska droga, t-b, 28 aug., leg. & det. mw; opu, blue tourist trail, around fort iv, mixed forest, 29 aug., leg. & det. gd. ciboria batschiana (zopf ) n.f. buchw. on acorns, tpu, t-c, 6 sep., leg. & det. gd. cudoniella acicularis (bull.) j. schröt. on stump of quercus robur l., gpu, t-c, 27 aug., leg. & det. js. #geoglossum umbratile sacc. on ground, among mosses, tpu, szuszalewo (0.2 km ne), peatbog, 12 sep., leg. ah, det. ms. notes. species known in poland from two localities: białowieża national park [9] and masurian landscape park [10]. helvella crispa fr. on ground, kopu, t-c, 7 sep., leg. & det. gd. helvella lacunosa afzel. on ground, opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. gd & ah, det. ak; r. helvella macropus (pers.) p. karst. on ground, gpu, deciduous forest, 27 aug., leg. & det. ms. helvella villosa (hedw.) dissing & nannf. on ground, opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. gd & ah, det. ak. humaria hemisphaerica (f.h. wigg.) fuckel on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. gd, det. ak; gpu, 27 aug. leg. & det. ms, opu, sośnia, 28 aug., garden, leg. & det. ak; opu, sośnia, 28 aug., mixed forest, leg. & det. mw. hymenoscyphus fructigenus (bull.) fr. on acorns, opu, osowiec-twierdza, t-c, 27 aug., leg. & det. dk; tpu, 6 sep., t-c, leg. gd, det. bg. hymenoscyphus salicellus (fr.) dennis on branch of salix fragilis l., bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. js. hymenoscyphus scutula (pers.) w. phillips on stem of plant, gpu, t-b, 27 aug., leg. & det. ms. mollisia discicolor (mont.) w. phillips on wood of quercus robur l., gpu, t-c, 27 aug., leg. &, det. ms. mollisia ramealis (p. karst.) p. karst. on branch of betula pendula roth, gpu, t-b, 27 aug., leg. &, det. js. orbilia xanthostigma (fr.) fr. on branch of deciduous tree, opu, sośnia, r-a, 28 aug., leg. &, det. js. otidea alutacea (pers.) massee on ground, gpu, deciduous forest, 27 aug., leg. &, det. ms. 4 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data otidea onotica (pers.) fuckel on ground, kpu, kapice, mixed forest, 30 aug., leg. &, det. ds; tpu, 6 sep., t-c, leg. gd, det. bg. peziza badia pers. on ground, opu, sośnia, pine forest, 28 aug., leg. &, det. ak. peziza fimeti (fuckel) seaver on deer (?) droppings, opu, sośnia, dune, 28 aug., leg. &, det. ak. peziza succosa berk. on ground, tpu, t-c, 6 sep., leg. gd, det. bg. basidiomycota agaricus arvensis schaeff. on ground, opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. gd, det. bg. #agaricus litoralis (wakef. & a. pearson) pilát on ground, opu, biały grąd educational path, sandy grasslands, 27 aug., leg. gd, det. bg. notes. species known in poland from a single locality [11]. #agaricus macrocarpus (f.h. møller) f.h. møller on ground, wpu, v-p, 8 sep., leg. gd, det. bg. notes. species known in poland from a few localities [12,13]. agaricus silvaticus shaeff. on ground, tpu, t-c, 6 sep., leg. gd, det. bg. agaricus silvicola (vittad.) peck on ground, tpu, t-c, 31 aug., leg. gd, det. bg. agrocybe firma (peck) kühner on litter, tpu, t-c, 31 aug., leg. gd, det. bg. amanita argentea huijsman on ground, opu, blue tourist trail, around fort iv, pine forest, 29 aug., leg. gd, det. bg. amanita fulva (schaeff.) pers. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg; gpu, t-b, 27 aug., leg. & det. ms; gpu, deciduous forest, 27 aug., leg. tl, det. ms; opu, carska droga, t-b, 28 aug., leg. & det. mw; opu, t-b, 2 sep., leg. & det. gd. amanita muscaria (l.) hook. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. ak. amanita phalloides (vaill.) link on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. mrm & nm, det. ak; opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. & det. md. amanita rubescens (pers.) gray on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak & ah, det. ak; opu, sośnia, pine forest, 28 aug., leg. jc, det. ak; opu, sośnia, t-b, 28 aug., leg. & det. mw. amphinema byssoides (pers.) j. erikss. on wood of pinus sylvestris l. and mooses, opu, sośnia, pine forest, 28 aug., leg. tl, det. tl&mp (ncbi accession no. ku525690). auriscalpium vulgare gray on cone of pinus sylvestris l., opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. & det. dk; gpu, t-c, 27 aug., leg. mrm, det. ak; gpu, dune, 27 aug., leg. & det. ds; opu, sośnia, pine forest, 28 aug., leg. & det. mw & ak. 5 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data baeospora myosura (fr.) singer on cone of picea abies (l.) h. karst., tpu, t-c, 6 sep., leg. gd, det. bg. bankera fuligineoalba (j.c. schmidt) pouzar on ground, opu, sośnia, pine forest, 28 aug., leg. bg & ah & ak, det. ak; e. bjerkandera adusta (willd.) p. karst. on wood of deciduous tree, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. gd & ah, det. ak. boletus edulis bull. on ground, opu, sośnia, pine and larch forest, 28 aug., leg. & det. bg & gd & dś & ak. bovista nigrescens pers. on ground, kpu, kapice, roadside, 30 aug, leg. & det. dś. bovista paludosa lév. on ground, tpu, t-c, 3 sep., leg. & det. gd; v, pp. bovista plumbea pers. on ground, tpu, szuszalewo (0.2 km ne), peatbog, 12 sep., leg. ah, det. ms. calocera cornea (batsch) fr. on wood of deciduous trees, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. mrm & nm, det. ak. calocera viscosa (pers.: fr.) fr. on wood of conifers, tpu, t-c, 31 aug. leg. & det. gd. calocybe carnea (bull.) donk on litter, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. ak; sośnia, lawn, 28 aug., leg. ct, det. bg. calvatia excipuliformis (scop.) perdeck on ground, bpu, meadow, 5 sep., leg. & det. gd. calvatia utriformis (bull.) jaap on ground, opu, sośnia, dune with pinus sylvestris l., 28 aug., leg. & det. ak. cantharellula umbonata (j.f. gmel.) singer on ground, gpu, 27 aug., leg. nm, det. ak. cantharellus cibarius fr. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. mw. ceriporia purpurea (fr.) donk on branch of fraxinus excelsior l., opu, sośnia, ecotone between mixed forest and alder forest, 28 aug., leg. & det. mw; e. ceriporia reticulata (hoffm.) domanski on branch of corylus avellana l., bpu, grobla honczarowska, r-a, 26 aug., leg.& det. mp (ncbi accession no. ku525691); r. chalciporus piperatus (bull.) bataille on ground, opu, sośnia, pine forest, 28 aug., leg. & det. ak. chroogomphus rutilus (schaeff.) o.k. mill. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. ak; opu, sośnia, dune with pinus sylvestris l., 28 aug., leg. & det. mw; opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 28 aug., leg. & det. mw. clavicorona pyxidata (pers.) doty on wood, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. tl & mp & lk, det. ak; opu, sośnia, pine forest, 28 aug., leg. mp, det. ak; opu, blue tourist trail, around fort iv, mixed forest, 29 aug., leg. & det. gd; góra skobla, t-c, 1 sep. leg. & det. gd; v. 6 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data clavulina cinerea (bull.) j. schröt. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. ak & ms; opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd & ah, det. ak; opu, sośnia, dune with pinus sylvestris l. and picea abies (l.) h. karst., 28 aug., leg. mw, det. bg. clavulina coralloides (l.) j. schröt. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. tl & mp & lk, det. ak. #clavulinopsis subtilis (pers.: fr.) corner among mosses, gpu, grzędy, t-b, 27 aug, leg. & det. ms. notes. species known in poland from one historical [5] and two contemporary localities [14]. clitocybe clavipes (pers.) p. kumm. on ground, opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. & det. dk; opu, sośnia, pine forest, 28 aug., leg. mrm, det. ak. clitocybe geotropa (bull.) quél. on ground, opu, sośnia, mixed forest, 28 aug., leg. & det. mw. clitocybe gibba (pers.) p. kumm. on ground and litter, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. bg, det. ak; opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. dk & md, det. ak; wpu, gugny, pine forest, 27 aug., leg. jc, det. ak. clitocybe odora (bull.) p. kumm. on ground, tpu, t-c, 6 sep., leg. & det. gd. clitopilus hobsoni (berk. & broome) p.d. orton on dead polyporaeae, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. ak. clitopilus prunulus (scop.) p. kumm. on ground, opu, sośnia, in adler forest edge, 28 aug., leg. mw, det. mw & bg. collybia cookei (bres.) j.d. arnold, on debris of fungi, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. mrm & nm, det. ak. coltricia perennis (l.) murrill on ground, opu, sośnia, pine forest, 28 aug., leg. mrm & ak & jc, det. mrm. conocybe albipes (g.h. otth.) hauskn. var. albipes, on ground, opu, biały grąd educational path, 27 aug., pasture, leg. ak, det. bg. conocybe ochrostriata hauskn. var. ochrostriata on ground, opu, biały grąd educational path, pasture, 27 aug., leg. & det. bg. notes. species not listed in checklist of polish larger macromycetes [5], however it was already reported from zakopane [15], and later collected in ujście warty national park (kujawa, ślusarczyk, unpublished), krosno town [16] and kampinos national park [14]. conocybe semiglobata (kühner) ex kühner & watling var. semiglobata on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg. conocybe siliginea (fr.) kühner on ground, opu, biały grąd educational path, pasture, 27 aug., leg. & det. bg. conocybe velutipes (velen.) hauskn. & svrček var. velutipes on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg. 7 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data #conocybe velutipes (velen.) hauskn. & svrček var. nitrophila hauskn. on ground, opu, biały grąd educational path, pasture, on cow dung, 27 aug., leg. ak, det. bg. notes. nitrophilous taxon, in poland known only from wrocław city (gierczyk, unpublished), rare in europe. coprinus disseminatus (pers.) quél. on stumps of deciduous trees, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg; opu, osowiec twierdza, lawn, 26 aug., leg. jc. det. bg; opu, biały grąd educational path, pasture, deciduous shrubs, 27 aug., leg. & det. mw; opu, sośnia, garden, 28 aug., leg. mw, det. bg. #coprinus heterosetulosus locq. on deer droppings, sośnia, peatbog, 28 aug., leg. gd, det. bg. notes. coprophilous species, not rare in poland but probably overlooked because of minute basidiocarps. reported from bieszczady mts and białowieża national park [17]. coprinus lagopus (fr.) fr. var. lagopus on ground, sośnia, r-a, 28 aug., leg. tl, det. bg. coprinus micaceus (bull.) fr. on wood, gpu, t-c, 28 aug., leg. a. bernatowicz, det. bg; opu, sośnia, garden, 28 aug., leg. mw, det. bg. coprinus plicatilis (curtis) fr. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg. #coprinus pseudoniveus bender & uljé on cow dungs, opu, biały grąd educational path, pasture, 27 aug., leg. & det. bg. notes. very rare, coprophilous species, in poland known only in bieszczady mts [17]. #coprinus pseudoradiatus kühner & joss. ex watling on deer droppings, opu, sośnia, peatbog, 28 aug., leg. gd, det. bg. notes. rather common species in poland, rarely collected due to minute basidiocarps. recorded only in the bieszczady mts [17] and białowieża primeval forest [18]. #coprinus pusillulus svrček (c. heptemerus m. lange & a.h. sm. var. parvisporus j. breitenb. & f. kränzl.) on deer droppings, sośnia, peatbog, 28 aug., leg. gd, det. bg. notes. coprophilous species, not rare in poland but probably overlooked due to minute basidiocarps. reported from the góry kaczawskie mts (in [17] as c. heptemerus var. parvisporus) and kampinos national park [14]. coprinus saccharinus romagn. stump of salix sp., bpu, carska droga near grobla honczarowska, roadside, 26 aug., leg. & det. bg. coprinus schroeterii p. karst. on ground, tpu, t-c, 31 aug., leg. gd, det. bg. coprinus stercoreus (scop.) fr. on deer droppings, opu, sośnia, spruce forest, 28 aug., leg. & det. bg; opu, sośnia, peatbog, 28 aug., leg. gd, det. bg. coprinus subimpatiens m. lange & a.h. sm. on litter, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. dś, det. bg. 8 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data #coprinus subpurpureus a.h. sm. on ground, opu, biały grąd educational path, pasture, 27 aug., leg. & det. bg. notes. very rare species, in poland known only from wrocław city [17]. coprinus xanthothrix romagn. on litter, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg; peat pits by the carska droga, 26 aug., leg. & det. bg; gpu, t-c, leg. mrm, det. bg. coriolopsis gallica (fr.) ryvarden on twigs of alnus sp., bpu, grobla honczarowska, 26 aug., leg. & det. ms; r. cortinarius armillatus (fr.) fr. on ground, opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 27 aug., leg. & det. mw. cortinarius cinnamomeus (l.) fr. on ground, opu, sośnia, mixed forest, 28 aug., leg. & det. mw. cortinarius collinitus (sowerby) gray on ground, opu, sośnia, pine forest, 28 aug., leg. & det. mw. cortinarius delibutus fr. on ground, gpu, mixed forest, leg. tl, det. ms. crepidotus cesati (rabenh.) sacc. var. subsphaerosporus (j.e. lange) senn-irlet on twigs of populus sp., opu, osowiec-twierdza, góra skobla, t-c, 1 sep., leg. gd, det. bg. crepidotus lundellii pilát on wood, opu, sośnia, pine fores margin, 28 aug., leg. gd, det. bg. crepidotus mollis (schaeff.) staude var. calolepis (fr.) pilát on wood of deciduous trees, opu, blue tourist trail, around fort iv, mixed forest, 29 aug., leg. & det. gd; opu, osowiec-twierdza, góra skobla, t-c, 1 sep., leg. gd, det. bg. crucibulum leave (huds.) kambly. on wood, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. bg, det. bg & ak; opu, sośnia, pine forest, 28 aug., leg. & det. ak. cylindrobasidium laeve (pers.) chamuris on branch of corylus avellana l., bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. ms. cystoderma amianthinum (scop.) fayod on litter, opu, sośnia, pine forest, 28 aug., leg. ak, det. bg. #cystoderma tuomikoskii harmaja on litter, opu, sośnia, spruce forest, 28 aug., leg. & det. bg; opu, sośnia, pine forest, 28 aug., leg. gd, det. bg. notes. very rare, boreal species, in poland hitherto known only from kampinos national park [14]. cystolepiota seminuda (lasch) bon on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg. daedalea quercina (l.) pers. on wood of quercus robur, tpu, t-c, 31 aug. leg. & det. gd. daedaleopsis confragosa (bolton) j. schröt. on wood, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak & ah, det. ak; opu, sośnia, r-a, 28 aug., leg. & det. js & mw; opu, osowiec-twierdza, góra skobla, r-a, 28 aug., leg. & det. 9 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data mw; park buffer zone, 2 km from osowiec-twierdza, peatbog, 28 aug., leg. & det. ms. disciseda bovista (klotzsch) henn. on ground, opu, sośnia, dune, 28 aug., leg. & det. ak; e. ditiola peziziformis (lév.) reid on branch of betula pendula roth, gpu, 27 aug., leg. dś, det. ak; e. entoloma caeruleum (p.d. orton) noordel. on ground, tpu, peatbog, 3 sep., leg. gd, det. bg. notes. rare but widespread blue entoloma species, growing in various habitats, including peat bogs and peaty grasslands. entoloma juncinum (kühner & romagn.) noordel. on ground, gpu, grzędy, deciduous forest near the tower; 27 aug., leg. & det. ms; r. #entoloma plebejoides (s.schulz.) noordel. on ground, tpu, pasture, 31 aug., leg. gd, det. bg. notes. rare species, in poland known only from poznań city (gierczyk, unpublished). entoloma rhodopolium (fr.) p. kumm. f. nidorosum (fr.) noordel. on ground, among mosses, gpu, grzędy, t-b, 27 aug., leg. & det. ms. entoloma sericeum (bull.) ex quél. on ground, at the road osowiec-grajewo, n from rudzki kanał, meadow, 26 aug., leg. gd, det. bg. notes. collected specimes represent probably untypical, pale form of var. cinereoopacum. exidia glandulosa (bull.) fr. on wood of deciduous trees, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. js; gpu, 27 aug., leg. & det. js. fistulina hepatica (schaeff.) with. at base of quercus robur l., opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. ak, det. bg & ak; tpu, t-c, 6 sep. leg. & det. gd; r, pp. flammulaster muricatus (fr.) watling on wood of deciduous trees, opu, blue tourist trail, around fort iv, mixed forest, 29 aug., leg. gd, det. bg; tpu, t-c, 6 sep., leg. gd, det. bg. flammulina fennae bas on wood, tpu, t-c, 31 aug., leg. & det. gd. fomes fomentarius (l.) j. kickx f. on log of deciduous tree and on betula pendula roth, gpu, t-c, 27 aug., leg. & det. js & ms; opu, sośnia, mixed forest, 28 aug., leg. & det. js & mw; opu, carska droga, t-b, 28 aug., leg. & det. mw; tpu, t-c, 31 aug., leg. & det. gd. fomitopsis pinicola (sw.) p. karst. on wood, gpu, t-c, 27 aug., leg. tl, det. ms; opu, carska droga, t-b, 28 aug., leg. & det. mw; opu, sośnia, r-a, 28 aug., leg. & det. js; opu, osowiec-twierdza, góra skobla, mixed forest, 30 aug., leg. & det. mw. galerina clavata (velen.) kühner on ground among mosses, opu, biały grąd educational path, pasture, 27 aug., leg. ak, det. bg; tpu, peatbog, 3 sep., leg. gd, det. bg. 10 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data galerina paludosa (fr.) kühner among sphagnum sp., gpu, t-b, 28 aug., leg. & det. ms; park buffer zone, 2 km from osowiec-twierdza, peatbog, 28 aug., leg. & det. ms; tpu, szuszalewo (0.2 km ne), peatbog, 12 sep., leg. ah, det. ms; r. galerina tibiicystis (atk.) kühner among sphagnum sp., park buffer zone, 2 km from osowiec-twierdza, peatbog, 28 aug., leg. & det. ms. galerina vittiformis (fr.) singer var. vittiformis among mosses, tpu, peatbog, 3 sep., leg. gd, det. bg. ganoderma applanatum (pers.) pat. on wood of deciduous trees, park buffer zone, 2 km from osowiec-twierdza, peatbog, 28 aug., leg. & det. ms; opu, osowiec-twierdza, góra skobla, 28 aug., leg. & det. mw; opu, osowiec-twierdza, headquarters of the park, 28 aug., leg. & det. jc. geastrum fimbriatum fr. on ground, opu, sośnia, dune, 28 aug., leg. mrm, det. ak; r. geastrum minimum schwein. on ground, opu, sośnia, dune, 28 aug., leg. & det. ak; e. geastrum pectinatum pers. on ground, opu, sośnia, dune, 28 aug., leg. jp, det. ak; wpu, v-p, 8 sep., leg. & det. gd; v. geastrum schmidelii vittad. on ground, opu, biały grąd educational path, dune and sandy grasslands, 27 aug., leg. gd & ah, det. ak; e, sp. geastrum striatum dc. on ground, opu, sośnia, garden, 28 aug., leg. & det. ak; e. gloeophyllum sepiarium (wulf.) p. karst. on wood, opu, sośnia, dune, 28 aug., leg. & det. mw & ms. gloeoporus dichrous (fr.) bres. on wood of deciduous trees, opu, t-b, 2 sep., leg. & det. gd; tpu, t-c, 31 aug., leg. & det. gd; e. gloeoporus taxicola (pers.) gilb. & ryvarden on log of pinus sylvestris l., opu, osowiec-twierdza, góra skobla, t-c, 1 sep., leg. & det. gd; r. #gloiodon strigosus (sw.) p. karst. on wood of salix sp., opu, sośnia, dune, 28 aug., leg. mp, det. bg; ex. notes. in poland known only in białowieża national park, very rare [19]. gomphidius glutinosus (schaeff.) fr. on ground, opu, sośnia, pine forest, 28 aug., leg. mrm & ms, det. ak & ms; opu, sośnia, dune, 28 aug., leg. & det. mw; r. gomphidius roseus (fr.) fr. on ground, sośnia, pine forest, 28 aug., leg. mrm, det. ak; r. gymnopilus penetrans (fr.) murrill on branch of corylus avellana l., opu, mixed forest, 9 sep., leg. gd, det. bg. gymnopilus picreus (pers.) p. karst. on log of pinus sylvestris l., opu, mixed forest, 9 sep., leg. gd, det. bg; e. gymnopus aquosus (bull.) antonín & noordel. on litter, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. gd, det. ak. 11 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data gymnopus confluens (pers.) antonín, halling & noordel. on litter, opu, osowiectwierdza, góra skobla, t-c, 27 aug., leg. md, det. ak; gpu, t-c, 27 aug., leg. mrm, det. ak; opu, sośnia, pine forest, 28 aug., leg. & det. mw & ms; opu, osowiectwierdza, góra skobla, mixed forest, 29 aug., leg. & det. mw. gymnopus dryophilus (bull.) murrill on litter, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. bg & ak, det. ak; gpu, deciduous forest, 27 aug., leg. mrm & ms, det. ak & ms. gymnopus peronatus (bolton) antonín, halling & noordel. on ground, gpu, t-c, 27 aug., leg. mrm, det. ak; opu, sośnia, pine forest, 28 aug., leg. gd, det. ak. hapalopilus nidulans (fr.) p. karst. on wood of deciduous trees, tpu, t-c, 31 aug., leg. & det. gd. hebeloma crustuliniforme (bull.) quél. on ground, opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. md, det. ak. #hebeloma velutipes bruchet on ground, tpu, t-b, 4 sep., leg. gd, det. bg. heterobasidion annosum (fr.) bref. s. l. on stump, opu, sośnia, pine forest, 28 aug., leg. & det. mw; opu, pine forest, 2 sep., leg. & det. gd. hygrocybe conica (schaeff.) p. kumm. var. conica on ground, opu, sośnia, dune, 28 aug., leg. & det. ak. hygrocybe conica (schaeff.) p. kumm. var. conicopalustris (r. haller aar.) heinem. on ground, opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. & det. mw; tpu, peatbog, 3 sep., leg. gd, det. bg. hygrophoropsis aurantiaca (wulfen) j. schröt. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. mrm; tpu, t-c, 31 aug., leg. & det. gd. hymenochaete rubiginosa (schrad.) lév. on branch of quercus robur l., gpu, t-c, 27 aug., leg. & det. js & ms; tpu, t-c, 31 aug., leg. & det. gd. hyphodontia paradoxa (schrad.) langer & vesterh. s. l. on wood, gpu, t-c, 27 aug., leg. & det. ms. inocybe appendiculata kühner on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. mrm, det. bg. #inocybe arenicola (r. heim) bon on ground, opu, sośnia, dune, 28 aug., leg. & det. bg. notes. rare species occurring in sandy habitats, e.g., inland dunes. in poland known only from białowieża primeval forest [18] and kampinos national park (in [20] as i. fastigiata var. arenicola). inocybe asterospora quél. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak, det. bg. inocybe calamistrata (fr.) gillet on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. tl & lk & mp, det. bg; v. inocybe calida velen. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak, det. bg; opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. & det. bg. 12 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data inocybe calospora quél. on ground, opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg; v. inocybe curvipes p. karst. on ground, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. & det. bg. #inocybe decipiens bres. on ground, opu, sośnia, dune, 28 aug., leg. & det. bg. notes. rare species occurring in sandy, calcareous habitats, in poland collected only once, in lower silesia [21]. inocybe dulcamara (pers.) p. kumm. on ground, opu, sośnia, dune, 28 aug., leg. & det. bg. #inocybe dunensis p.d. orton on ground, gpu, dune, 27 aug., leg. tl & lk & mp, det. bg; opu, sośnia, dune, 28 aug., leg. & det. bg. notes. species growing on sandy dunes under salix, in poland known only from słowiński national park [22]. inocybe fuscidula velen. opu, sośnia, dune, 28 aug., leg. & det. bg. inocybe geophylla (fr.) p. kumm. var. geophylla on ground, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. bg & gd, det. bg; gpu, dune, 27 aug., leg. & det. ms; opu, sośnia, pine forest, 28 aug., leg. mrm & ms, det. bg & ms; opu, sośnia, r-a, 28 aug., leg. & det. ak; opu, sośnia, mixed forest, 28 aug., leg. & det. mw; park buffer zone, between osowiec and białogrądy, 30 aug., meadow, leg. gd, det. bg. inocybe grammata quél. on ground, opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. & det. bg; v. #inocybe inodora velen. on ground, opu, sośnia, dune, 28 aug., leg. & det. bg. notes. rare species hitherto known only from the świętokrzyskie province [23]. inocybe lacera (fr.) p. kumm. var. lacera on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg; opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg; opu, sośnia, pine forest, 28 aug., leg. jc, det. bg; opu, sośnia, dune, 28 aug., leg. ak, det. bg. inocybe leptophylla g.f. atk. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak, det. bg. inocybe maculata boud. on ground, opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg. #inocybe microspora j.e. lange on ground, on ground, opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg. notes. uncommon species, hitherto known only form kampinos national park [14] and vicinity of poznań (gierczyk, unpublished), growing in deciduous and mixed forests. inocybe mixtilis (britzelm.) sacc. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak, det. bg; sośnia, dune, 28 aug., leg. ak, det. bg. #inocybe mystica stangl & glowinski (i. cryptocystis d.e. stuntz) on ground, opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg. 13 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data notes. very rare species, occurring in deciduous and mixed forests on nutrient-rich soils. in poland reported only from the kielce city [24]. #inocybe oblectabilis (britzelm.) sacc. on ground, opu, carska droga, roadside, 26 aug., leg. ak, det. bg. notes. rare species preferring sandy soils, in poland known only from the babia góra mt [25] and łódź province [26]. #inocybe obscurobadia (j. favre) grund & d.e. stuntz on ground, opu, sośnia, dune, 28 aug., leg. gd, det. bg. notes. rare species, in poland known only from kampinos national park [14]. inocybe ochroalba bruyl. on ground, opu, sośnia, dune, 28 aug., leg. gd, det. bg. notes. rare species, hitherto collected at the babia góra mt [27], in białowieża primeval forest [18], kampinos national park [14] and radomsko district (gierczyk, nowicki, unpublished). inocybe perlata (cooke) sacc. on ground, tpu, t-c, 31 aug., leg. gd, det. bg. #inocybe pseudoasterospora kühner & boursier var. microsperma kuyper & p.-j. keizer on ground, opu, sośnia, spruce forest, 28 aug., leg. ak & bg, det. bg. notes. not rare but probably overlooked species, growing under deciduous and picea trees. in poland known from kampinos national park [14] and vicinity of radziechowice village [28]. inocybe rennyi (berk. & broome) sacc. on ground, opu, sośnia, dune, 28 aug., leg. gd, det. bg. notes. rare species growing on sandy soils and mossy heathlands with coniferous trees. #inocybe serotina peck on ground, opu, sośnia, dune, 28 aug., leg. gd, det. bg. notes. rare species growing on sandy dunes under deciduous and coniferous trees. in poland found only at inland dunes in kampinos national park [20]. #inocybe subnudipes kühner on ground, opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg. notes. species known from the świętokrzyskie province [23] and kampinos national park [14]. inocybe vulpinella bruyl. on ground, opu, sośnia, dune, 28 aug., leg. & det. bg. notes. rare species growing at dunes and in other sandy habitats. inonotus obliquus (pers.) pilát on betula pendula roth, opu, sośnia, deciduous forest, 28 aug., leg. & det. gd; r, pp. inonotus radiatus (sowerby) p. karst. on alnus glutinosa gaertn., opu, sośnia, r-a, 28 aug., leg. & det. mw. 14 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data inonotus tomentosus (fr.) teng on pinus sylvestris l., opu, blue tourist trail, around fort iv, pine forest, 29 aug., leg. & det.gd; v. irpex ochraceus (pers.) kotir. & saaren. (steccherinum ochraceum (pers.) gray) on wood of deciduous tree, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. gd & mw & ms, det. bg & ms; opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. & det. mw; osowiec-twierdza, headquarters of the park, 28 aug., leg. & det. jc. laccaria laccata (scop.) berk. & broome on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. ak & ms; opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. md, det. ak; gpu, t-b, 27 aug., leg. & det. ms; carska droga, t-b, 28 aug., leg. & det. mw; tpu, peatbog, 3 sep., leg. gd, det. bg; tpu, szuszalewo (0.2 km ne), peatbog, 12 aug., leg. ah, det. ms. laccaria laccata (scop.) berk. & broome var. pallidifolia (peck) peck on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. mrm & nm, det. ak. notes. very common but rarely recognized variety, probably much widely occurring than nominative one (var. laccata). in poland reported from the bieszczady mts [29], ochojec reserve [30], kampinos national park [14] and already recorded in the biebrza national park [1]. laccaria proxima (boud.) pat. on ground, opu, t-b, 2 sep., leg. & det. gd. laccaria tortilis (bolton) cooke on ground, park buffer zone, near osowiec, meadow, 30 aug., leg. & det. gd. lactarius camphoratus fr. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. ak; gpu, deciduous forest, 27 aug., leg. & det. js. lactarius deterrimus gröger on ground, tpu, t-c, 6 sep., leg. & det. gd. lactarius glyciosmus (fr.) fr. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. ak. lactarius helvus (fr.) fr. on ground, opu, carska droga, t-b, 26 aug., leg. bg, det. ak. lactarius obscuratus (lasch) fr. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. gd, det. bg. lactarius pubescens (schrad.) fr. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. ak. lactarius pyrogalus (bull.) fr. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. gd, det. bg. lactarius quietus (fr.) fr. on ground, gpu, deciduous forest, 27 aug., leg. & det. ms. lactarius rufus (scop.) fr. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. ak. #lactarius scoticus berk. & broome on ground, tpu, peatbog, 3 sep., leg. gd, det. bg; tpu, t-b, 4 sep., leg. gd, det. bg. notes. rare species growing in peat bogs under betula trees. very rare in poland, reported only in rybojady reserve [31] and vicinity of nowy dworek village [32]. 15 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data lactarius torminosus (schaeff.) pers. on ground, sośnia, pine forest, 28 aug., leg. & det. gd. lactarius vietus (fr.) fr. on ground, opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 28 aug., leg. & det. mw. leccinum melaneum (smotl.) pilát & dermek on ground, opu, carska droga, t-b, 26 aug., leg. bg, det. ak. leccinum niveum (fr.) rauschert on ground, opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 28 aug., leg. & det. mw; opu, t-b, 2 sep., leg. & det. gd; v. leccinum pseudoscabrum (kallenb.) šutara on ground, opu, sośnia, mixed forest, 28 aug., leg. mw, det. ms. leccinum scabrum (bull.) gray on ground, gpu, deciduous forest, 27 aug., leg. tl, det. ms; park buffer zone, 2 km from osowiec-twierdza, peatbog, 28 aug., leg. & det. ms; opu, sośnia, pine forest, 28 aug., leg. & det. ak; opu, sośnia, mixed forest, 28 aug., leg. & det. ak; opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 28 aug., leg. & det. mw. leccinum variicolor watling on ground, opu, t-b, 2 sep., leg. & det. gd. leccinum versipelle (fr.) snell. on ground, opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 28 aug., leg. & det. mw. lentinellus ursinus (fr.) kühner on wood, gpu, deciduous forest, 27 aug., leg. & det. js; v. lentinus tigrinus (bull.) fr. on wood, opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd & ah, det. gd; r. lenzites betulinus (l.) fr. on wood, opu, sośnia, pine forest, 28 aug., leg. & det. gd; opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 28 aug., leg. & det. mw. lepiota alba (bres.) sacc. on ground, gpu, t-c, 27 aug., leg. mrm, det. bg; v. lepiota castanea quél. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak, det. bg. lepiota clypeolaria (bull.) p. kumm. on ground, tpu, t-c, 6 sep., leg. gd, det. bg. #lepiota cortinarius j.e. lange on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. tl & lk & mp, det. bg. notes. rare species, the nominative variety hitherto unreported in poland. the variety audreae has been collected in białowieża national park [33]. lepiota cristata (bolton) p. kumm. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg; opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg; sośnia, garden, 28 aug., leg. & det. gd; tpu, t-c, 3 sep., leg. gd, det. bg. lepiota ventriosospora d.a. reid on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg; opu, sośnia, roadside, 28 aug., leg. gd, det. bg; 16 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data tpu, t-c, 6 sep., leg. gd, det. bg; tpu, mixed forest, 29 aug., leg. gd, det. bg; opu, blue tourist trail, around fort iv, mixed forest, 29 aug., leg. & det. gd. lepista gilva (pers.) pat. on ground, gpu, t-c, 27 aug., leg. & det. ms. lycoperdon lividum pers. on ground, opu, sośnia, dune, 28 aug., leg. & det. mw; opu, osowiec-twierdza, headquarters of the park, lawn, 29 aug., leg. & det. mw. lycoperdon molle pers. on ground, opu, sośnia, dune, 28 aug., leg. & det. ms. lycoperdon nigrescens (pers.) pers. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. ak; opu, sośnia, dune, 28 aug., leg. & det. mw; tpu, t-c, 6 sep., leg. & det. gd. lycoperdon pyriforme schaeff. on wood, opu, sośnia, pine forest, 28 aug., leg. & det. mw. lyophyllum palustre (peck) singer among sphagnum sp., opu, carska droga, t-b, 26 aug., leg. gd, det. ak; park buffer zone, osowiec-twierdza 2 km, peatbog, 28 aug., leg. & det. ms; v. macrolepiota excoriata (schaeff.) wasser on ground, opu, biały grąd educational path, sandy grasslands, 27 aug., leg. gd, det. bg. macrolepiota mastoidea (fr.) singer on ground, gpu, deciduous forest, 27 aug., leg. ms, det. bg. macrolepiota nympharum (kalchbr.) wasser on ground, tpu, t-c, 31 aug., leg. gd, det. bg. macrolepiota olivieri (barla) wasser on ground, tpu, t-c, 31 aug., leg. & det. gd. macrolepiota procera (scop.) singer f. procera on ground, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. & det. gd & ah; opu, osowiectwierdza, góra skobla, t-c, 27 aug., leg. md, det. ak; opu, sośnia, meadow, 28 aug., leg. & det. ak & bg; opu, sośnia, near dune, 28 aug., leg. & det. mw. macrolepiota procera (scop.) singer f. permixta (barla) vizzini & contu on ground, opu, sośnia, dune, 28 aug., leg. & det. bg. notes. rather common taxon, known from many scattered localities in poland [33,34]. macrolepiota rhacodes (vittad.) singer on ground, gpu, deciduous forest, 27 aug., leg. tl, det. ms; wpu, v-p, 8 sep., leg. gd, det. bg. macrolepiota rhodosperma (p.d. orton) migl. var. rhodosperma (macrolepiota fuliginosa sensu vellinga) on ground, opu, blue tourist trail, around fort iv, mixed forest, 29 aug., leg. gd, det. bg. notes. not rare but rarely reported species, probably misidentified and listed as m. procera. known from few localities in poland [33]. marasmius oreades (bolton) fr. on ground, opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. & det. ak; opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. dk, det. ak; gpu, dune, 27 aug., leg. ms & tl & mp & lk, det. ms & ak. 17 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data marasmius scorodonius (fr.) fr. on ground, opu, sośnia, dune, 28 aug., leg. & det. ak. marasmius wynnei berk. & broome on litter, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg. megacollybia platyphylla (pers.) kotl. & pouzar on stumps of deciduous tree, gpu, t-c, 27 aug., leg. mrm, det. ak. #melanoleuca friesii (bres.) bon on ground, tapu, meadow, 4 sep., leg. gd, det. bg. notes. in poland known from kampinos national park [14] and białowieża primeval forest [18]. melanoleuca strictipes (p. karst.) murrill on ground, bpu, meadow, 5 sep., leg. gd, det. bg. merismodes anomalus (pers.) singer on fallen twigs of deciduous trees, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. tl & mp & lk, det. ak. mycena acicula (schaeff.) p. kumm. on fallen twigs of deciduous trees, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. bg & gd, det. bg. #mycena bulbosa (cejp) kühner the remnants of grass, opu, sośnia, deciduous forest, 28 aug., leg. gd, det. ak. notes. in poland known only from niepołomicka forest [5] and kampinos national park [14]. mycena galericulata (scop.) gray on stump and log of deciduous tree, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak & gd & ah, det. ak; opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. gd &a h, det. ak; gpu, deciduous forest, 27 aug., leg. & det. js. mycena haematopus (pers.) p. kumm. var. haematopus on log of deciduous tree, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. ms; opu, sośnia, r-a, 28 aug., leg. mw & tl, det. mw & ak; tpu, t-c, 6 sep., leg. & det. gd. mycena inclinata (fr.) quél. on wood, tpu, t-c, 6 sep., leg. gd, det. bg. mycena niveipes murrill on log of deciduous tree, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. mrm & nm & bg, det. ak. mycena pura (pers.) p. kumm. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. ak & bg; opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. md, det. ak; gpu, t-c, 27 aug., leg. mrm, det. ak; gpu, deciduous forest, 27 aug., leg. & det. ms; opu, sośnia, pine forest, 28 aug., leg. mrm & js & mw, det. mrm & mw; opu, carska droga, t-b, 28 aug., leg. & det. mw. mycena sanguinolenta (alb. & schwein.) p. kumm. on litter, gpu, deciduous forest, 27 aug., leg. nm, det. ak; gpu, t-b, 27 aug., leg. & det. ms. naucoria salicis p.d. orton on ground, park buffer zone, near osowiec, meadow, 30 aug., leg. gd, det. bg. omphalina sphagnicola (berk.) m.m. moser – among sphagnum sp., tpu, szuszalewo (0.2 km ne), peatbog, 12 sep., leg. ah, det. ms; v. 18 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data omphalina umbellifera (l.) quél. among sphagnum sp., park buffer zone, 2 km from osowiec-twierdza, peatbog, 28 aug., leg. & det. ms; r. panaeolus acuminatus (schaeff.) quél. on ground, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg. panaeolus alcidis m.m. moser on deer droppings, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. af-s, det. bg; gpu, t-c, 27 aug., leg. mrm, det. bg; opu, sośnia, dune, 28 aug., leg. & det. bg; sośnia, spruce forest, 28 aug., leg. & det. bg; opu, t-b, 2 sep., leg. gd, det. bg; opu, pine forest, 2 sep., leg. gd, det. bg. panaeolus foenisecii (pers.) kühner on ground, opu, biały grąd educational path, pasture, 27 aug., leg. ak, det. bg. panaeolus papilionaceus (bull.) quél. var. papilionacues on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg; opu, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg; r. panaeolus subbalteatus (berk. & broome) sacc. [p. cintulus (bolton) sacc.] on ground, opu, biały grąd educational path, pasture, 27 aug., leg. bg & ak, det. bg. panellus stipticus (bull.) p. karst. on wood, opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. gd & ah, det. ak. paxillus atrotomentosus (batsch) fr. on wood, opu, sośnia, pine forest, 28 aug., leg. & det. mrm. paxillus involutus (batsch) fr. s. l. on ground, wpu, gugny, pine forest, 27 aug., leg. jc, det. ak; opu, sośnia, pine forest, 28 aug., leg. & det. gd; opu, sośnia, mixed forest, 28 aug., leg. & det. mw. peniophora quercina (pers.) cooke on wood, opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. & det. mw. perenniporia medulla-panis (jacq.) donk on wood, tpu, t-c, 6 sep., leg. & det. gd; v. phallus impudicus l. on ground, tpu, t-c, 31 aug., leg. & det. gd. phanerochaete gigantea (fr.) s.s. rattan on wood, opu, sośnia, pine forest, 28 aug., leg. & det. mw. phellinus igniarius (l.) quél. on salix sp., bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak & ah & ms, det. ak & ms; opu, biały grąd educational path, oak forest on the mineral hill, 27 aug., leg. & det. mw; gpu, t-b, 27 aug., leg. & det. ms. phellinus robustus (p. karst.) bourdot & galzin (1928) on wood of quercus robur, tpu, t-c, 31 aug., leg. & det. gd. phellodon confluens (pers.) pouzar on ground, opu, sośnia, pine forest, 28 aug., leg. ak, det. bg; ex notes. species assumed to be extinct in poland but recorded from few contemporary localities [35] that should influence its red-list status in future. phlebia tremellosa (schrad.) nakasone & burds. on wood of deciduous trees, tpu, t-c, 6 sep., leg. & det. gd. 19 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data pholiota adiposa (fr.) p. kumm. on salix sp., bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. gd, det. bg; r. pholiota flammans (batsch) p. kumm. on wood, opu, sośnia, pine forest, 28 aug., leg. tl & bg & ak, det. ak & bg; osowiec-twierdza, góra skobla, t-c, 1 sep., leg. & det. gd. pholiota pityrodes (f. brig.) gröger on litter, biały grąd educational path, deciduous shrubs, 27 aug., leg. gd, det. bg. notes. rare species of poorly known distribution. hitherto mentioned from italy, denmark and germany [36]. growing on rhizomes of grasses, mainly phragmites communis. pholiota spumosa (fr.) singer on log of pinus sylvestris l., opu, pine forest, 9 sep., leg. gd, det. bg. pholiota squarrosa (weigel) p. kumm. at the base of populus sp., gpu, t-c, 29 aug., leg. & det. gd. pholiotina utricystidiata enderle & h.-j. hübner no ground, opu, biały grąd educational path, pasture, 27 aug., leg. & det. bg. notes. rather rare, annulate pholiotina species, reported from nordic countries, germany, the czech republic, and austria [37]. piptoporus betulinus (bull.) p. karst. on betula pendula roth, opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. & det. md; wpu, gugny, pine forest, 27 aug., leg. jc, det. ak; gpu, t-c, 27 aug., leg. & det. js & ms; opu, carska droga, t-b, 28 aug., leg. & det. mw; opu, sośnia, t-b, 28 aug., leg. & det. mw. pleurotus pulmonarius (fr.) quél. on branch of padus sp., opu, osowiec-twierdza, góra skobla, t-c, 1 sep., leg. gd, det. bg; v. pluteus atricapillus (batsch) fayod on wood of deciduous trees, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. bg, det. ak; opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. dk, det. ak; opu, dziekciarka, close to road 668 (from osowiec to sośnia), t-b, 27 aug., leg. & det. mw; gpu, t-c, 27 aug., leg. nm & ms, det. ak & ms; opu, sośnia, pine forest, 28 aug., leg. & det. ak. pluteus atromarginatus (singer) kühner on wood of pinus sylvestris l., osowiectwierdza, near park, square, 26 aug., leg. gd, det. ak; opu, sośnia, pine forest, 28 aug., leg. & det. ak. pluteus leoninus (schaeff.) p. kumm. on log of deciduous tree, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. mrm & nm, det. ak; opu, sośnia, deciduous forest, 28 aug., leg. gd, det. ak; opu, sośnia, pine forest, 28 aug., leg. & det. mw. pluteus nothopellitus justo & m.l. castro on wood of deciduous tree, sośnia, r-a, 28 aug., leg. mw, det. bg. notes. species not listed in checklist of polish larger macromycetes [5], already known from a few localities [35]. pluteus phlebophorus (ditmar) p. kumm. on wood, tpu, t-c, 6 sep., leg. gd, det. bg. 20 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data pluteus romellii (britzelm.) sacc. on fallen twigs of deciduous tree, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg; opu, osowiec -twierdza, góra skobla, t-c, 27 aug., leg. dk, det. ak. pluteus salicinus (pers.) p. kumm. on wood of deciduous trees, opu, sośnia, r-a, 28 aug., leg. tl, det. ak; bpu, meadow, 5 sep., leg. & det. gd; tpu, t-c, 6 sep., leg. gd, det. bg. polyporus arcularius (batsch) fr. on wood of deciduous tree, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg bg, det. ak; gpu, t-c, 27 aug., leg. mrm & tl, det. ak & ms; opu, sośnia, r-a, 28 aug., leg. & det. mw. polyporus badius (pers.) schwein. on log of salix fragilis l., opu, osowiec-twierdza, kanał rudzki, 26 aug., leg gd, det. ak. polyporus ciliatus fr. on wood of deciduous tree, opu, sośnia, r-a, 28 aug., leg. & det. mw. polyporus melanopus (pers.) fr. on wood of deciduous tree, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg ak & ah, det. ak; e. polyporus tuberaster (jacq.) fr. on wood, gpu, t-c, 27 aug., leg. tl, det. ms; r. polyporus varius (pers.) fr. on wood, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg tl & mp & lk, det. ak; gpu, t-c, 27 aug., leg. tl, det. ms. psathyrella candolleana (fr.) maire on litter, opu, sośnia, spruce forest, 28 aug., leg. & det. bg. psathyrella populina (britzelm.) kits van wav. on wood, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg ak, det. bg; e. psathyrella prona (fr.) gillet on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg & det. bg. psathyrella pseudocorrugis (romagn.) bon ss. kits van wav., arnolds, enderle on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg & det. bg; biały grąd educational path, deciduous shrubs, 27 aug., leg. & det. bg. notes. common species, recently collected also in the kampinos national park [14], beskid niski mts [37], vicinity of przemyśl, radomsko district and knyszyńska primeval forest (gierczyk, unpublished). pseudohydnum gelatinosum (scop.) p. karst. on wood of picea abies (l.) h. karst., opu, sośnia, pine forest, 28 aug., leg. & det. mrm. pseudomerulius aureus (fr.) jülich on wood of pinus sylvestris l., opu, osowiectwierdza, góra skobla, t-c, 1 sep., leg. & det. gd; r. #pseudotomentella griseopergamacea m.j. larsen on wood, opu, sośnia, r-a, 28 aug., leg. tl, det. mp (ncbi accession no. ku525692). notes. species not listed in checklist of polish larger macromycetes [5], recently reported from a single locality [39]. psilocybe elongata (pers.) j.e. lange [hypholoma elongatum (pers.) ricken] among sphagnum sp., gpu, peatbog, 27 aug., leg. & det. js; park buffer zone, osowiec-twierdza 2 km, peatbog, 28 aug., leg. & det. ms; opu, t-b, 2 sep., leg. gd, det. bg; r. 21 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data psilocybe fascicularis (huds.) noordel. [hypholoma fasciculare (huds.) p. kumm. var. fasciculare] on wood, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. bg; opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. dk & mw, det. mw & ak; gpu, t-c, 27 aug., leg. & det. js & ms; opu, sośnia, pine forest, 28 aug., leg. & det. ak. psilocybe lateritia (schaeff.) noordel. [hypholoma sublateritium (fr.) quél.] on wood, opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. & det. mw. psilocybe squamosa (pers.) p.d. orton [stropharia squamosa (pers.) quél.] on ground, tpu, t-c, 6 sep., leg. gd, det. bg; i. psilocybe uda (pers.) gillet [hypholoma udum (pers.) quél.] among sphagnum sp., park buffer zone, 2 km from osowiec-twierdza,peatbog, 28 aug., leg. & det. ms; opu, t-b, 2 sep., leg. gd, det. bg; r. pycnoporellus fulgens (fr.) donk on pinus sylvestris l., wpu, v-p, 8 sep., leg. & det. gd; v. ramaria eumorpha (p. karst.) corner on ground, opu, mixed forest, 2 sep., leg. gd, det. bg; osowiec-twierdza, góra skobla, t-c, 1 sep., leg. gd, det. bg. ramaria gracilis (pers.) quél. on ground, opu, sośnia, dune, 28 aug., leg. tl & mp & lk, det. bg; e. ramaria stricta (pers.) quél. on ground, gpu, deciduous forest, 27 aug., leg. & det. ms. #ramaria suecica (fr.) donk on ground, opu, sośnia, dune, 28 aug., leg. mw, det. bg. notes. rare species, known from two historical [5] and two contemporary localities [40,41]. resupinatus trichotis (pers.) singer on wood of corylus avellana l., bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg & det. ak. rhizopogon roseolus (corda) th. fr., on ground, opu, sośnia, dune, 28 aug., leg. & det. ds. rhodocollybia maculata (alb. & schwein.) singer on ground, opu, sośnia, pine forest, 28 aug., leg. mrm & mw, det. ak & mw. rickenella fibula (bull.) raithelh. on stump among mosses, opu, sośnia, dune, 28 aug., leg. & det. ak; opu, sośnia, pine forest, 28 aug., leg. & det. mw; gpu, t-b, 29 aug., leg. & det. ms; bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg ak & ah, det. ak; tpu, peatbog, 3 sep., leg gd, det. bg. rickenella setipes (fr.) raithelh. on wood of deciduous trees among mosses; gpu, t-b, 29 aug., leg. & det. ms. ripartites tricholoma (alb. & schwein.) p. karst. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg af-s, det. ak. rozites caperatus (pers.) p. karst. on ground; opu, sośnia, pine forest, 28 aug., leg. & det. mrm & mw. russula aeruginea lindbl. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. mw. 22 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data russula claroflava grove on ground under betula pendula roth, gpu, t-b, 27 aug., leg. mrm & ms, det. ak & ms. russula cyanoxantha (schaeff.) fr. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. mw. russula emetica (schaeff.) pers. on ground, opu, sośnia, pine forest, 28 aug., leg. & det. mw. russula ochroleuca (pers.) fr. on ground, gpu, deciduous forest, 27 aug., leg. & det. ms; opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 28 aug., leg. & det. mw; opu, carska droga, t-b, 28 aug., leg. & det. mw. russula pectinatoides peck on ground, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. & det. mw. russula xerampelina (schaeff.) fr. on ground, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. gd & ah & mw, det. ms & mw. schizophyllum commune fr. on log and branch of deciduous tree; bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. & det. ak & ms; opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. gd & ah, det. ak. scleroderma areolatum ehrenb. on ground, opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. dk, det. ak; opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. gd & ah, det. ak. scleroderma bovista fr. on ground, opu, sośnia, garden, 28 aug., leg. & det. ak; tpu, t-c, 6 sep., leg. gd, det. bg; opu, t-c, 9 sep., leg. gd, det. bg. scleroderma cepa pers. on ground, sośnia, near the forest, 28 aug., leg. & det. js; e. scleroderma citrinum pers. on ground, carska droga, t-b, 28 aug., leg. & det. mw; park buffer zone, 2 km from osowiec-twierdza, peatbog, 28 aug., leg. & det. ms. stereum hirsutum (willd.) gray on wood of deciduous trees, gpu, deciduous forest, 27 aug., leg. & det. ms; opu, sośnia, r-a, 28 aug., leg. & det. js; opu, osowiectwierdza, góra skobla, mixed forest, 28 aug., leg. & det. mw. stropharia alcis kytöv. on deer droppings, opu, sośnia, spruce forest, 28 aug., leg. & det. bg. suillus bovinus (l.) roussel, on ground, opu, osowiec-twierdza, góra skobla, mixed forest, 28 aug., leg. & det. mw. suillus granulatus (l.) roussel on ground; opu, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. & det. dk; gpu, deciduous forest, 27 aug., leg. ds, det. ak; opu, sośnia, pine forest, 28 aug., leg. ak & bg & jc, det. ak & bg; opu, sośnia, dune, 28 aug., leg. & det. mw. suillus luteus (l.) roussel on ground, opu, sośnia, pine forest, 28 aug., leg. mrm & jc, det. mrm; opu, sośnia, dune, 28 aug., leg. & det. mw. suillus variegatus (sw.) kuntze on ground, opu, sośnia, pine forest, 28 aug., leg. & det. ak. thelephora caryophyllea (schaeff.) fr. on ground, opu, sośnia, dune, 28 aug., leg. & det. ak & mw & ms & tl; v. 23 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data thelephora palmata (scop.) fr. on ground, osowiec-twierdza, góra skobla, t-c, 27 aug., leg. dk, det. ak. thelephora penicillata (pers.) fr. on ground, tpu, t-c, 31 aug., leg. & det. gd; v. thelephora terrestris ehrh ex willd. on litter and ground, park buffer zone, osowiectwierdza 2 km, peatbog, 28 aug., leg. & det. ms; opu, t-b, 2 sep., leg. gd, det. bg. #tomentella atramentaria rostr. on wood, bpu, grobla honczarowska, r-a, 26 aug., leg. & det. mp (ncbi accession no. ku525693); opu, sośnia, r-a, 28 aug., leg. & det. mp (ncbi accession no. ku525694). notes. a single historical locality was reported (probably incorrectly) in the polish literature [42]. tomentella badia (link) stalpers on wood of salix sp., opu, sośnia, r-a, 28 aug., leg. & det. mp (ncbi accession no. ku525695). notes. species not listed in checklist of polish larger macromycetes [5], reported later in the contemporary literature [43]. tomentella ellisii (sacc.) jülich & staplers on wood, uroczysko dęby forest complex, t-c, 27 aug., leg. & det. mp (ncbi accession no. ku525696). notes. species not listed in checklist of polish larger macromycetes [5], reported in the contemporary literature from a few localities [35]. tomentella lapida (pers.) stalpers on wood, bpu, grobla honczarowska, r-a, 26 aug., leg. & det. mp (ncbi accession no. ku525697). notes. species not listed in checklist of polish larger macromycetes [5], reported in the contemporary literature from a few localities [35]. tomentella lateritia pat. on wood, opu, sośnia, r-a, 28 aug., leg. tl, det. mp (ncbi accession no. ku525698). tomentella pilosa (burt) bourdot & galzin on wood, bpu, grobla honczarowska, r-a, 26 aug., leg. & det. mp (ncbi accession no. ku525699). tomentella punicea (alb. & schwein.) j. shröt. on wood, uroczysko dęby forest complex, on the border between forest and meadow, 27 aug., leg. & det. mp (ncbi accession no. ku525700). tomentella sublilacina (ellis & holw.) wakef. on wood, uroczysko dęby forest complex, on the border between forest and meadow, 27 aug., leg. & det. mp (ncbi accession no. ku525701). trametes gibbosa (pers.) fr. on wood, opu, osowiec-twierdza, góra skobla, t-c, 28 aug., leg. & det. mw; opu, osowiec-twierdza, headquarters of the park, 28 aug., leg. & det. jc; opu, opu, sośnia, mixed forest, 28 aug., leg. & det. jc. trametes hirsuta (wulfen) pilát on stump of deciduous tree, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. ak & ah, det. ak; opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. md, det. ak; opu, sośnia, pine forest, 28 aug., leg. jc, det. ak; opu, sośnia, mixed forest, 28 aug., leg. & det. mw. trametes ochracea (pers.) gilb. & ryarden on wood, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. gd & ah, det. ak. 24 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data trametes versicolor (l.) pilát on wood, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. gd & ms, det. ak & ms; opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. & det. md; opu, sośnia, pine forest, 28 aug., leg. jc, det. ak; opu, sośnia, mixed forest, 28 aug., leg. & det. jc. tremella mesenterica retz. on twigs of betula pendula roth; bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. gd, det. ak. trichaptum fuscoviolaceum (ehrenb.) ryvarden on wood of coniferous trees, opu, sośnia, pine forest, 28 aug., leg. & det. ak. tricholoma lascivum (fr.) gillet on ground, opu, sośnia, mixed forest, 28 aug., leg. & det. mw. tulostoma brumale pers. on ground, opu, osowiec-twierdza, square at the park, 26 aug., leg. gd, det. ak; r. tulostoma kotlabae pouzar on ground, opu, sośnia, dune, 28 aug., leg. bg & cw, det. ak; e, sp. notes. protected species, recorded in poland in few localities [5,13,44,45]. vascellum pratense (pers.) kreisel on ground, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. af-s, det. ak. xerocomus badius (fr.) kühner ex e.-j. gilbert on ground, opu, sośnia, pine forest, 28 aug., leg. jc & mw, det. ak & mw; opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 28 aug., leg. & det. mw. xerocomus pascuus (pers.) krombh. s. l. on ground, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. md & mw, det. ak & mw; opu, dziekciarka, close to road 668 (from osowiec to sośnia), mixed forest, 27 aug., leg. & det. mw. xerocomus porosporus imler on ground, opu, biały grąd educational path, oak forest on mineral hill, 27 aug., leg. af-s, det. ak. xerocomus rubellus (krombh.) quél. on ground, bpu, grobla honczarowska, deciduous shrubs, 26 aug., leg. mrm & nm, det. bg. xerocomus subtomentosus (l.) quél. on ground, opu, sośnia, deciduous forest, 28 aug., leg. & det. gd. xeromphalina cauticinalis (fr.) kühner & maire var. subfellea bon on litter, opu, sośnia, pine forest, 28 aug., leg. gd, det. bg. discussion the high number of macrofungi (336 species) recorded during this relatively short and fragmentary survey in the biebrza national park, clearly demonstrated the high mycological potential of this site. in terms of species richness (483 species) our data from two few-days surveys, conducted in late summer of 2012 and 2013 are similar to the results of two-year, full season investigation in the wigry national park (516 species) [46]. so far less than 500 species was recorded from other polish national parks: woliński (approximately 460 species; stasińska, sotek, unpublished), świętokrzyski (about 450) [47], słowiński (429) [22], poleski (418) [40], bory tucholskie (413) [48], drawieński (379; stefaniak, unpublished), magurski (340) [49] and karkonoski (200) [50]. this 25 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data simple comparison of the numbers of species reported from different national parks should be treated with caution as they are diverse in terms of habitat and plant species richness. when compared to our results, the numbers listed above suggest that the knowledge of macromycetes occurring in polish national parks is poor in most cases, and that the patterns of macrofungi species richness in poland must be considered as insufficiently recognized. it is worthy to highlight that among the fungi collected in bbnp are taxa very rare in poland and europe. some species were found in poland only in the bbnp. the presented data are also valuable in terms of fungal conservation and threat assessment, as many new localities of the fungi endangered in poland are given. particularly important is the finding of a new locality of gloiodon strigosus – the species very rare in europe [51] that was assumed to be extinct in poland. the only contemporary locality of the species in poland comes from the early twenty-first century from the białowieża national park. the knowledge of macrofungi species diversity in national parks, particularly of very rare and endangered species distribution is necessary for an effective protection of the most vulnerable components of polish mycobiota. summarizing, based on research to date (including data from present survey and those reported previously [1,52] and collected during fourth congress of european mycologists [53]) 508 macrofungispecies have been recorded in bbnp. the need of the extended studies in that protected and highly diverse area is recommended in order to assess mycobiota of bbnp in relation to the various habitats represented in the park. key outcomes ■ eight species, hitherho not reported in any publication from poland were found in bbnp in 2013: conocybe velutipes var. nitrophila, entoloma caeruleum, e. plebejoides, inocybe rennyi, i. vulpinella, pholiota pityrodes, pholiotina utricystidiata, and tomentella pilosa. ■ seven taxa protected by polish law [54] are known to occur in the bbpn: bovista paludosa, fistulina hepatica, ganoderma lucidum, geastrum schmidelii, inonotus obliquus, tulostoma kotlabae, and xerocomus parasiticus. ■ the data gathered in bbnp extend the knowledge of chorology and ecology of 95 red-listed taxa: ex – gloiodon strigosus, phellodon confluens; e – bankera fuligineoalba, ceriporia purpurea, disciseda bovista, ditiola peziziformis, geastrum minimum, g. rufescens, g. schmidelii, gloeoporus dichrous, gymnopilus picreus, hydnellum concrescens, phallus duplicatus, phleogena faginea, pholiota albocrenulata, p. squarrosoides, polyporus melanopus, psathyrella populina, pseudomerulius aureus, ramaria gracilis, scleroderma cepa, tulostoma kotlabae; v – bovista paludosa, clavicorona pyxidata, geastrum pectinatum, inocybe calamistrata, i. calospora, i. grammata, inonotus tomentosus, ishnoderma benzoinum, leccinum niveum, lentinellus ursinus, lepiota alba, lyophyllum palustre, mycena megaspora, omphalina sphagnicola, perenniporia medulla-panis, phyllotopsis nidulans, pleuroybella porrigens, pleurotus pulmonarius, pycnoporellus fulgens, thelephora caryophyllea, t. penicillata, xylobolus frustulatus; r – antrodiella serpula, asterophora lycoperdoides, calocera furcata, ceriporia reticulata, clavariadelphus fistulosus, climacocystis borealis, clitocybe cf. agrestis, c. hydrogramma, coprinus niveus, coriolopsis gallica, diplomitoporus flavescens, entoloma juncinum, exidia glandulosa, fistulina hepatica, galerina paludosa, ganoderma lucidum, geastrum fimbriatum, gloeoporus taxicola, gomphidius glutinosus, g. roseus, gyroporus castaneus, g. cyanescens, helvella lacunosa, inonotus obliquus, lactarius trivialis, lentinus tigrinus, lentinus torulosus, leucocortinarius bulbiger, oligoporus ptychogaster, omphalina umbelifera, onygena equuina, panaeolus papilionaceus, paxillus rubicundulus, phellinus pini, pholiota adiposa, physisporinus vitreus, polyporus tuberaster, psilocybe elongatum, p. udum, pynoporus cinnabarinus, sparassis crispa, trichaptum biforme, tulostoma brumale; i – cystoderma granulosum, hygrophorus hyphotejus var. hyphotejus, hygrophorus hyphotejus var. aureus, lentaria byssiseda, mycena 26 of 28© the author(s) 2016 published by polish botanical society acta mycol 50(2):1070 kujawa et al. / macromycetes in biebrza national park – new data peliathina, pluteus plautus, psilocybe squamosa, tremella foliacea, tricholoma equestre. our results give only preliminary insight into the biodiversity of fungal component of ecosystems at the biebrza national park. however, this inventory is an important contribution to the knowledge of mycobiota of the areas protected in poland. the localities of selected species of highest conservation priority should be monitored with the support of the state. acknowledgments we are deeply grateful to director of the biebrza national park, mr. roman skapski as well as to mr. andrzej grygoruk, and other employees of the biebrza national park for collecting fungi, offering their facilities, helping in the implementation of our research, and creating the very friendly atmosphere during our visit. references 1. kujawa a, wrzosek m, domian g, kędra k, szkodzik j, rudawska m, et al. preliminary studies of fungi in the biebrza national park (ne poland). ii. macromycetes. acta mycol. 2012;47(2):235–264. http://dx.doi.org/10.5586/am.2012.027 2. ruszkiewicz-michalska m, wrzosek m, tkaczuk c, dynowska m, sucharzewska e, szkodzik j. preliminary studies of fungi in the biebrza national park. part i. micromycetes. acta mycol. 2012;47(2):213–234. http://dx.doi.org/10.5586/am.2012.026 3. ruszkiewicz-michalska m, bałazy s, chełkowski j, dynowska m, pawłowska j, sucharzewska e, et al. preliminary studies of fungi in the biebrza national park (ne poland). part iii. micromycetes – new data. acta mycol. 2015;50(2):1067. http://dx.doi.org/10.5586/ am.1067 4. matuszkiewicz w. przewodnik do oznaczania zbiorowisk roślinnych polski. warszawa: pwn; 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rozporządzenie ministra środowiska z dnia 9 października 2014 r. w sprawie ochrony gatunkowej grzybów. dz. u. z 2014 r., poz. 1408. http://www.grzyby.pl/rejestr-grzybow-chronionych-i-zagrozonych.htm http://dx.doi.org/10.1007/s00572-011-0387-x http://dx.doi.org/10.1007/s00572-011-0387-x http://dx.doi.org/10.5586/am.2011.011 http://dx.doi.org/10.5586/am.2011.011 http://dx.doi.org/10.5586/am.1058 http://dx.doi.org/10.5586/am.1968.011 abstract introduction material and methods results list of species collected in 2013 discussion key outcomes acknowledgments references 2016-01-29t11:36:41+0000 piotr otręba rhizoctonia solani ag5 and its offspring – morphology and sensitivity to fungicides patrycja hendel* 0000-0002-4346-97281, ewa moliszewska 0000-0003-4919-51391, małgorzata nabrdalik 0000-0002-2565-90341, paweł kudrys 0000-0003-1776-48941, natalia knap1 1 university of opole, faculty of natural sciences and technology, institute of environmental engineering and biotechnology, opole, poland * to whom correspondence should be addressed: patrycja.hendel@uni.opole.pl abstract the objective of these studies was to identify differences and similarities within the progeny of rhizoctonia solani ag5, which arose from basidiospores produced by the maternal strain id23. the following characteristics were analyzed: appearance of the mycelium (color, structure, zonation, and presence of sclerotia), growth rate (at 10°c, 20°c, and 30°c), fungicide sensitivity, and hyphal structure. the mycelial color of r. solani ag5 ranged from white/cream to light and dark brown. the structure of the mycelium may be compacted or flattened with visible zoning or fluffy with dark brown sclerotia on the colony surface. homokaryons and heterokaryons derived from homokaryons were analyzed by constructing a phylogenetic tree using morphological data. single basidiospore-grown isolates formed a separate subclade, the majority of which were grouped with a maternal isolate; however, heterokaryons derived from them created a separate subclade. in addition, isolates grown in basidiospores germinated at low temperatures created their own group, but with some exceptions. this shows a divergence in the morphological parameters of the subsequent generations and within generations. the optimal temperature for growth was found to be between 20°c and 30°c. the exceptions were strains obtained from basidiospores that germinated at refrigerated temperatures. for these samples, 10°c was found to be the optimal growth temperature. the hyphae of homokaryons were characterized by the presence of branching at an almost right angle and the presence of a septum at the site of constriction of the branch itself. the mean diameter of hyphae ranged 2.93–15.60 μm, depending on the age of hyphae. the fungicidal compounds at a concentration of 10 ppm had no significant effect on the activity of the tested strains, whereas a tenfold increase in the dose reduced the growth ability of the tested isolates. the activity of fungicides containing azoxystrobin, thiuram, or thiophanate-methyl on r. solani resulted in a reduction in the mycelial growth rate only in the case of azoxystrobin and thiuram, and in some cases, it was completely inhibited (thiophanate-methyl). po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 1 mailto:patrycja.hendel@uni.opole.pl keywords: rhizoctonia solani, basidiospore, progeny, morphology, hyphae, fungicides, azoxystrobin, thiuram, thiophanate-methyl. po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 2 introduction rhizoctonia solani belongs to phylum basidiomycota. this pathogen is widely distributed worldwide. it has a wide host range and attacks more than 250 plant species. this fungus attacks the root systems, stems, seedlings, fruits, and leaves. in the natural environment, they occur in the form of mycelial hyphae. r. solani can survive in the form of sclerotia for up to several years in soil or plant debris (nagrodzka et al., 2016). the mycelium produced by r. solani is colorless in the initial stage of development and ranges from light to dark brown in later stages. it consists of cells with an elongated shape and numerous branches. each cell is tapered at the end and has a transverse septum located near the junction with the neighboring cell (moliszewska et al., 2002; stępniewska-jarosz, 2012). to date, 13 anastomosis groups (ag) have been identified within r. solani. these groups were further subdivided into subgroups. the reason for this subdivision is the great heterogeneity of r. solani and the ability of hyphae to join. ag2 and ag4 are the most frequently described groups because of their high pathogenicity in various plant species (stępniewska-jarosz, 2006). this study focuses on the little-studied group ag5, which is considered to have low plant pathogenicity. it can be found on sugar beet seedlings with symptoms of seedling damping-off, among others. this group has not yet been fully characterized and described, and progeny production has not been developed under laboratory conditions. the aim of this study was to describe the morphological profile of the progeny obtained from basidiospores of one ag5 field strain. materials and methods isolates. the isolate r. solani id23 was used in this study, as the maternal strain was isolated in 1997 from diseased sugar beet seedlings with symptoms of damping-off. seedlings were collected from a field area close to opole, poland. all fungi (table 1) used in this study belonged to the r. solani collection created by e. moliszewska at the university of opole, institute of environmental engineering and biotechnology, and are routinely stored on pda slants in a refrigerator (for usual use) and on barley grains overgrown by mycelium in a deep freezer for long-term storage. species determination for id23 was performed using classical anastomosis tests, which showed that this strain belongs to ag5. this was confirmed by isolation and sequencing of the its1-its2 fragment of ribosomal dna. the sequences were compared to those in the genbank database (moliszewska, 2009). recently, this sequence was confirmed using the blast tool in the ncbi database. po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 3 table 1. list of tested r. solani strains (various colors indicate sbis obtained in separate cycles of sexual fruiting) no. single basidiospore isolates (sbi) heterokaryons (h) 1 sbi17a id23 i 13 h23 ii ii 2 sbi17b id23 ii 15 h23 ii iii 1 3 sbi25 id23 iii 19 h23 ii iv 4 id23 – 2015 a id23 v 25 h23 ii iv 1 5 id23 – 2015 b id23 vi 27 h23 ii vii 6 id23 – 2015 c id23 viii 28 h23 ii viii 7 id23 – 2017 i id23 ix 30 h23 iii ix 1 8 id23 x 33 h23 iii ix 2 9 17l h23 v x 1 10 18l h23 v x 2 11 24l 12 31l 13 34l perfect stage culturing basidiospore collecting. this study was based on a group of isolates obtained by culturing r. solani ag5 basidiospores under laboratory conditions. all cultured basidiospores were collected from r. solani ag5-labeled id23 during several cycles of sexual fruiting. initially, basidiospore production and collection were performed for a few isolates of the ag5 group using the soil-over method described by toda and hyakumachi (2006) (photo 1). finally, fruiting was obtained for only one isolate, which was coded as id23. basidiospores were trapped on water agar (wa) and cultured at approximately 20℃. each mycelium developed from a basidiospore was transferred to fresh pda medium. wa plates with no germinating basidiospores were placed in a refrigerator and incubated at approximately 10°c for the next two months, and then newly developed mycelia were transferred to fresh pda medium. the letter ‘l’ has been added in the lab code for the mycelia developed in 10°c. po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 4 photo 1. perfect stages of rhizoctonia solani. left fruiting visible on the soil surface. right basidium and basidiospore of rhizoctonia solani dyed with safranine 0; (×400) heterokaryon formation. single basidiospore isolates (sbis) served as progenies and parental strains to produce potential heterokaryons (hs). potential hs were obtained through cultural pairing of sbis (parental strains) and their cleavage from the contact sites of the two paired strains. strains were inoculated on the opposite sides of the petri dish on pda medium with 2% activated carbon. after the distinctive tuft was created, potential hs were transplanted onto a new pda medium. morphology. the progenies of two generations (sbis and hs) were analyzed in terms of their morphological diversity. for the purpose of this study, all strains were inoculated onto pda medium and incubated at 10°c, 20°c, and 30°c in the dark. the daily mycelial growth rate was observed for each strain. after two weeks of incubation, mycelia were morphologically characterized (color, concentric zonation, mycelial height and fluffiness, compactness, and presence of sclerotia). the experiment was performed with four replicates. morphological features of the analyzed fungi were compared by creating a similarity/dissimilarity tree using ntsyspc 2.21w software and the upgma method. fungicides. in the next phase of the research, pda supplemented with fungicides, azoxystrobin, thiophanate-methyl, and thiuram, used at concentrations of 10 and 100 ppm, allowed for evaluation of the resistance of the mycelium to these fungicides. plates with pda without supplements served as controls. in both experiments, the growth of mycelia [mm] was measured every 24 h along two vertical lines on the bottom of the petri dishes for all tested 2 μm po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 5 samples until the mycelium covered the entire dish (ɸ 90 mm). the fungicide test was performed at 21 ± 2°c. all experiments were performed in triplicate. the results of measuring mycelial growth are expressed as the range of mycelial increases per day (mm/d). based on the data obtained from the test with fungicides, the coefficient of growth inhibition was calculated according to abbot’s formula (1925): ∆h=𝐾𝐾0−𝐹𝐹 𝐾𝐾0 ∙ 100% where ∆h is the coefficient of linear growth inhibition of the fungus [%], k0 is the diameter of the colony on the control dish [mm], and f is the colony diameter on the dish with fungicide. in the final stage of the study, microscopic measurements of the samples incubated at 20°c were performed. the average hyphae diameter, as well as the minimum and maximum diameters [µm], were measured at a magnification of ×400. data analysis. the standard deviations were calculated for the growth parameters obtained in the tests using excel. statistical analysis was performed using the anova method (p = 0.05) using the statistica program, and significant differences among groups were compared by tukey’s test. results isolates – culturing, identification, heterokaryons, and monokaryons. the basic classification of the id23 isolate and dna isolation was performed in a previous study by moliszewska (2009). maternal isolate id23 was confirmed to be r. solani ag5. the sequence showed 99.86% identity with the r. solani ag5 strain (accession no. jq946291) and 100% identity with thanathephorus cucumeris strain gz-5 was determined to be ag5 (accession no. kr006028), both of which were deposited in genbank. the id23 sequence was submitted to the ncbi genbank database under accession no. op601638. strains were incubated at room temperature for two weeks and after which morphological characteristics were macroscopically evaluated, starting with the parental isolate. based on this evaluation, progeny were further evaluated. maternal r. solani id23 was characterized by a light brown fluffy structure. irregularly shaped and brown sclerotia were visible on the surface of the mycelium. zonation of mycelial growth was not observed on the surface of the maternal id23 isolate. the color of the tested strains ranged from light beige to light or dark brown. individual isolates also differed in the mycelial fluffiness, height, and hyphae density. in a few cases, concentric zonation and sclerotia have also been observed. unfortunately, distinction of the po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 6 progenies by color to distinguish individual basidiospore-born isolates and hs is not possible. however, they were grouped separately in a dendrogram of similarity (figure 1). po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 7 figure 1. similarity tree based on morphological features of maternal strain id23 and its progenies. strains with letter “h” in a code are heterokaryons po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 8 in each offspring group, mycelia took on different hues. some strains developed a light beige to almost white mycelium, whereas others had a dark brown color. however, most isolates were light brown in color. concentric zonation within the cultures was observed in strains whose mycelium grew flat and did not form sclerotia on their surfaces. in the observed isolates, all tested hs were found to have the ability to form sclerotia as compact mycelial structures. they appear in the form of brownand creamy-colored densely packed structures. these structures were also characterized by irregular or almost spherical shapes (table 2). r. solani ag5 isolates are characterized by their very diverse mycelial appearance. depending on the strain, they can vary in terms of fluffiness and height. the study showed that high and fluffy mycelia were formed by all potentially heterokaryotic isolates and a few single basidiospore isolates. the remaining r. solani strains obtained from single basidiospores grew flat and formed compact structures (table 2). table 2. morphological characteristics of r. solani ag5 isolates characteristics model picture sbi heterokaryons c ol or w hi te / b ei ge 18l, 17l, sbi 17a, sbi 17b, 27, 25, sbi25, 15, 13, 28, 19, id23-2017-i, id23-2015a, id23 ii, id23-2015b, id23-2015c h23-ii-ii, h23-iiiii-1 l ig ht b ro w n 31l, 30, 33, id23 vi, id23 x, id23 ix, id23 viii, id23 iii, id23 i h23-v-x-1, h23v-x-2, h23-iii-ix1, h23-iii-ix-2, h23-ii-viii d ar k br ow n id23 v, 24l, 34l h23-ii-iv, h23-ii-iv-1, h23-ii-vii, po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 9 st ru ct ur e fl at 18l, 17l, sbi 17a, sbi 17b, 27, 25, sbi25, 15, 13, 33, 30, 28, 19, id23 vi, id23 x, id23 ix, id23 viii, id23 v, id23 iii, id23 i, id23-2017-i, id23-2015a, id23 ii, id23-2015b, id23-2015c f lu ff y 24l, 31l, 34l h23-ii-iii-1, h23v-x-1, h23-v-x2, h23-iii-ix-1, h23-iii-ix-2, h23ii-viii, h23-ii-vii, h23-ii-iv, h23-iiiv-1, h23-ii-ii sc le ro ti a cu rr en t id23 ix, id23 x, 19, 34l, id23 vi, id23 v, 33, id23 iii, id23 viii h23-ii-iii-1, h23v-x-1, h23-v-x2, h23-iii-ix-1, h23-iii-ix-2, h23ii-viii, h23-ii-vii, h23-ii-iv, h23-iiiv-1, h23-ii-ii ab se nt 18l, 17l, sbi17a, sbi17b, sbi25, 31l, 24l, 27, 25, 15, 13, 30, 28, id23-2017-i, id232015a, id23-2015b, id23-2015c, id23 ii, id23 i z on in g c ur re nt 18l, 17l, sbi 17a, sbi 17b, 27, 25, sbi25, 15, 13, 33, 30, 28, 19, id23 vi, id23 x, id23 ix, id23 viii, id23 v, id23 iii, id23 i ab se nt 24l, 31l, 34l,id232017-i, id23-2015a, h23-ii-iii-1, h23v-x-1, h23-v-x2, h23-iii-ix-1, h23-iii-ix-2, h23po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 10 id23 ii, id23-2015b, id23-2015c ii-viii, h23-ii-vii, h23-ii-iv, h23-iiiv-1, h23-ii-ii based on the calculations, the daily growth at 10°c was determined to range from 2.5 to 15.2 mm/d, with the maternal strain being 4.6 mm/d (figure 2). all other isolates, except a few strains signed with the letter “l,” had a similar daily growth, oscillating between the values of 4.5–6.5 mm/d. the sbi isolates labeled “l” were characterized by the following growth values: 18l 13.5, 17l 14.8, 34l 12.5, 31l 15.2, and 24l 14.7 mm/d, indicating that these samples grew most favorably under these temperature conditions and their growth rate was much higher than that of the maternal strain id23. when hs were compared with their id23 parent isolates, their growth rate was found to be inconsiderable different from that of the parent, ranging 3.2–6.4 mm/d. at a temperature of 20°c, the daily growth rate for the mother strain id23 was 14.2 mm/d and the maximum value for the isolates studied was 21.4 mm/d (isolate id23 ii), and the minimum value was 5.7 mm/d (isolate 15) (figure 2). increasing the incubation temperature to 30°c did not have as drastic effect on the modification of mycelial growth rates as when the temperature was increased from 10°c to 20°c. the daily growth of the maternal strain id23 under these conditions was 14.1 mm/d. the highest recorded value was 21 mm/d (isolate h23-v-x-2), whereas the lowest was 6 mm/d (isolate 18l), which clearly confirmed the tendency of the isolates to have different optimal growth temperatures. at higher temperatures, the growth rate did not change over a wide range, and in most of the tested strains, it was similar. the lowest values of daily growth at 30°c were observed for “l” labeled-isolates. hs at this temperature showed slightly higher growth rates than the parental strains. only r. solani h23-ii-viii grew like one of its parent strains id23 viii (17.8 mm/d). for the other hs, the growth rates were 0.2–2.9 mm/d higher at 30°c than at lower temperatures (figure 2). the graph in figure 2 shows comparison of the strains tested at all incubation temperatures. based on our observations, the most favorable temperature for rapid mycelium growth was concluded to be 30°c. however, the values observed in the tests conducted at 20°c suggest that a temperature range of 20°c–30°c may be optimal for the growth of r. solani. at a low temperature (10°c), the growth rates of the strains tested were less effective, as these isolates required a much longer incubation period. however, “l” labeled-isolates (obtained po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 11 after the basidiospore incubation in the refrigerator) were the only ones that showed more favorable growth at 10°c. this confirms that these isolates did not spontaneously activate their growth at room temperature, which was too high for them, and transferring the isolation wa plates to the cold room allowed them to grow at the beginning of this study. statistical analysis (p=0.05) showed that the growth of the tested groups of fungi (maternal isolate, sbi, and hs) did not differ inconsiderable at 10°c, although considerable differences were observed between sbi and hs at 20°c and 30°c. maternal isolates did not differ notable in either group of progeny. po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 12 figure 2. influence of temperature on the average daily growth (mm/d) of tested mycelia ag5 group of r. solani 0 5 10 15 20 25 30 id23 sbi17a sbi17b sbi25 13 15 17l 18l 19 24l 25 27 28 30 31l 33 34l id23 – 2017 i id23 – 2015 a id23 – 2015 b id23 – 2015 c id23 i id23 ii id23 iii id23 v id23 vi id23 viii id23 ix id23 x h23 ii ii h23 ii iii 1 h23 ii iv h23 ii iv 1 h23 ii vii h23 ii viii h23 iii ix 1 h23 iii ix 2 h23 v x 1 h23 v x 2 daily growth [mm/d] st ra in 30℃ 20 ℃ 10℃ po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 13 microscopic evaluation of the examined strains of r. solani ag5. microscopic evaluation of the mycelia was performed to determine the hyphal diameter (µm), considering the minimum, maximum, and average values. maternal strain id23, five exemplary potential hs, and 12 single basidiospore isolates were considered in this test. in terms of anatomical structure, the hyphae of the progeny of r. solani ag5 were found to have diameters ranging 2.93 μm to 15.60 μm. the mean size of the hyphae in potential hs ranged 3.90–13.65 μm. in the parent strain id23, the mean value was 6.14 μm, the minimum was 3.90 μm, and the maximum was 7.80 μm. for sbi, these values were 2.93–15.60 μm. this could mean that single basidiospore isolates present greater differences in the diameter of the hyphae (figure 3). analysis of the similarity between the hs and their parent strains revealed that the maximum size of hyphae of isolate h23-ii-iii was the same as of its parent strains (9.75 μm); however, the mean value of the measurements was higher than that of isolate id23 ii, but identical to that of id23 iii. isolate h23-ii-viii had a larger diameter than its parent strain id23 ii; however, it was similar as that of the second parental strain id23 viii (figure 3). another potential heterokaryon, h23-iii-ix-2, resembled its parental isolate id23 ix in the minimum and maximum diameter of hyphae, whereas the overall average size, in this case, was smaller than that of both parental strains. the maximum diameter of the r. solani h23-v-x-1 hyphae was identical to that of id23 v and id23 x, and the minimum diameter was the same as that of id23 v, whereas a greater similarity was observed in the mean value of this parameter with r. solani id23 x. general analysis of the relationship based on the diameter of the young hyphae of r. solani ag5 showed that the hs did differ inconsiderablefrom the parental strains in their morphological structure. in the analysis, in most cases, the progeny was observed to adopt the characteristics of one of the parents or have the features of both, on average. po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 14 figure 3. hyphal diameter of selected monokaryons and heterokaryons of r. solani ag5 (µm) fungicides. two fungicide concentrations were used in this study: 10 ppm of active substance, which refers to the possible field concentration of the preparations, and 100 ppm of active substance, which is extremely high; therefore, sensitive isolates should be extracted. on the medium containing 10 ppm of azoxystrobin (amistar), the growth of the mycelium was found to be inhibited by 29% (isolate id23 vi); however, in the case of the maternal strain id23 and isolates sbi 17a, 24l, and id23 viii, it did not contribute to the slowing down of mycelial growth (figure 4). on the 10 ppm medium with thiuram, some isolates grew unchanged, which means that the fungicide had no effect on mycelium growth (isolates id23, sbi17a, and id23 2017-i), and the remaining strains slowed down their growth to a greater or lesser extent. the calculated growth inhibition coefficient (δh) varied between 0%–35%, and in one strain, the value was as high as 89% (id23 i) (figure 4). a slight slowdown in mycelial growth was observed on pda medium containing 10 ppm thiophanate-methyl. the calculated growth inhibition coefficients (δh) varied between 1%–43%, with low values for isolates id23, id23 ii, and id23 2017-i and the highest value for r. solani isolate 28 (figure 4). increasing the concentration of the active substance to 100 ppm resulted in increased growth inhibition of r. solani ag5 isolates (figure 5). on the pda medium with azoxystrobin, the growth rate of the tested fungi was found to be inhibited by 0%–66%, therefore, it was inhibited, in most cases, almost twice as effectively as at a lower fungicide concentration. the lowest value of growth inhibition was obtained for the single basidiospore isolate 24l and the 0 2 4 6 8 10 12 14 16 18 id 23 h 23 ii ii h 23 ii – ii i 1 h 23 ii v ii i h 23 ii i ix 2 h 23 v x 1 33 25 18 l 19 id 23 ii id 23 v id 23 v ii i id 23 x id 23 ix id 23 ii i sb i 2 5 sb i 1 7a h yp ha l d ia m et [ μm ] strains po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 15 highest value for a potential hs isolate h23-ii-viii (figure 4). the presence of thiuram contributed to an increase in growth inhibition compared to previous formulations, which in this case was 23%–80%. thiuram had the least effect on the single basidiospore isolate sbi17a, while the greatest effect was observed on potential hs, both strains h23-iii–ix-1 and h23-iii– ix-2 (figure 5). thiophanate-methyl (100 ppm) affected tested strains the most effectively. in this case, the inhibition of the growth rate varied between 3.5 mm/d (isolate sbi 17a) and 19.0 mm/d (isolate id23 v) (figure 4). for “cool” strains, 17l, 18l, 24l, 31l, 34l, and all potential hs, 100% growth inhibition by thiophanate-methyl was observed (figure 4). statistical analysis (p = 0.05) showed that significant differences among the tested groups of fungi (maternal isolate, sbi, and hs) did not exist with the use of 10 ppm fungicides. in the case of the 100 ppm fungicide concentration, differences were significant between sbi and hs, and the maternal isolate did not differ significantly from either group of strains. po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 16 figure 4. influence of selected fungicides (10 ppm) on the growth of individual isolates of r. solani ag5 showed as an inhibition coefficient (δh%) (pda+a azoxystrobin, pda+m thiophanate-methyl, and pda+t thiuram) 0 10 20 30 40 50 60 70 80 90 100 id23 18l 17l sbi 17b sbi 17a 34l 31l 24l 27 25 sbi 25 15 13 33 30 28 19 id23 vi id23 2017 i id23 2015 a id23 x id23 ix id23 viii id23 v id23 iii id23 ii id23 i id23 2015 c id 23 2015 b h23 ii iii 1 h23 v x 2 h23 v x 1 h23 iii ix 1 h23 iii ix 2 h23 ii viii h23 ii vii h23 ii iv 1 h23 ii iv h23 ii ii growth inhibition coefficient δh[%] st ra in pda+m pda+t pda+a po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 17 figure 5. influence of selected fungicides (100 ppm) on the growth of individual isolates of r. solani ag5 showed as an inhibition coefficient (δh%) (pda+aazoxystrobin, pda+m thiophanate-methyl, and pda+tthiuram). 0 10 20 30 40 50 60 70 80 90 100 110 id23 18l 17l sbi 17b sbi 17a 34l 31l 24l 27 25 sbi 25 15 13 33 30 28 19 id23 vi id23 2017 i id23 2015 a id23 x id23 ix id23 viii id23 v id23 iii id23 ii id23 i id23 2015 c id 23 2015 b h23 ii iii 1 h23 v x 2 h23 v x 1 h23 iii ix 1 h23 iii ix 2 h23 ii viii h23 ii vii h23 ii iv 1 h23 ii iv h23 ii ii growth inhibition coefficient δh [%] st ra in pda+m pda+t pda+a po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 18 discussion isolates of r. solani ag 5 derived from one maternal strain, id23, presented very different morphological features. the color of the mycelia ranged from almost white to creamy and light to dark brown. the mycelial structure varied from compact, with visible zonation, to fluffy, with distinct sclerotia on the colony surface. they appeared only on the edge of petri dishes during the initial phase of growth or developed on the surface of the entire colony. carling and sumner (1995) did not provide a morphological description of ag5; however, they detailed basic information on its pathogenicity, concluding that ag5 is far less virulent than ag3 and that it is thiamine auxotrophic. the morphology of rhizoctonia mycelia is usually given in general descriptions of subsequent anastomosis groups. for example, carling et al. (2002) showed that mycelia of rhizoctonia solani ag13 were characterized by a light to dark brown color. after a few days of incubation, they noticed zonation in all incubated samples, which disappeared over time. initially, dark brown sclerotia were observed only near the edge of the petri dishes, but after a longer cultivation period, they also appeared randomly scattered over the entire surface of the mycelium. moliszewska et al. (2020) described two ag11 isolates that formed white-beigeto creamy-colored mycelia with wide concentric zonation, and one of them formed light-colored sclerotia. this confirmed the morphological differences in the anastomosis group. morphological differences among ag5 isolates isolated from carrots in sweden were also observed by marcou et al. (2021). singh et al. (2018) observed the characteristics of r. solani isolated from seven agroecological zones that were adapted to maize cultivation in 2018. all 62 strains that were incubated were light to dark brown in color, and their structures varied from fluffy to compact. the studies on the growth of r. solani mycelia presented in this paper have shown that all isolates may grow more or less effectively depending on the ambient temperature. at 10°c, the average daily growth rate of mycelia was the lowest, ranging 1.9– 15.2 mm. at 20°c, the growth increase for the tested isolates varied between 6.4–21.4 mm/d. increasing the temperature to 30°c did not radically affect the growth activity of the fungal isolates compared to that at 20°c; in this case, the average growth of the mycelia per day varied between 6.0–21.0 mm. moliszewska et al. (2020) showed that for two polish ag11 isolates, the average daily rate of hyphal growth on pda at 21°c was 22.8 mm and 22.6 mm, respectively. the analysis of optimal growth temperature showed that r. solani ag5 grew the most favorably at both temperatures (20°c and 30°c), except the group of isolates, which showed po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 19 the highest growth activity at 10°c. this group of isolates was derived from single basidiospores incubated on wa in a refrigerator, which shows how great possibilities are coded in the r. solani ag5 genome in terms of optimal growth temperatures. this flexibility proves that it is always possible for some offspring to survive, independent of the environmental conditions. bełka and mańka (2014) conducted tests with different strains of r. solani (including ag5) and showed that ag5 isolates reached the fastest growth among other anastomosis groups (13.9 mm/d). furthermore, carling et al. (2002) showed that the average daily mycelial growth for r. solani (ag13) was approximately 28.3 mm at the temperature of 25°c. this was much higher than the growth rates of our tested ag5 isolates and those observed by bełka and mańka (2014) and moliszewska et al. (2020). tomaso-peterson (2007) identified pathogenic fungi in agronomic crops and turfgrasses, among which, 12 r. solani isolates were identified. one of them belonged to ag5, and its optimal growth temperature was found to be 21°c–29°c. harikrishnan et al. (2004) showed a relationship between temperature and the growth rate of r. solani. mycelial growth was observed in different anastomosis groups, including 15 isolates of ag5. although there were differences in growth rates among the tested isolates, in most cases, the fungi showed more effective growth with increasing temperature. none of the tested isolates showed the ability to grow below 5°c. the most rapid growth was recorded at temperatures of 20°c, 25°c, and 30°c, which agrees with our observations. papavizas and davey (1961) found that the optimum temperature that affects the saprotrophic activity of r. solani depends on the soil type. for example, in a greenhouse where loamy sand was used, the optimum temperature for mycelial activity was 20°c, and at 30°c, the efficiency of the saprotrophic mycelial development decreased. they also demonstrated that r. solani showed the highest saprobic activity in fine sand in the temperature range of 26°c– 30°c. similar observations have been reported by martinson (1963). their research showed that the activity of r. solani mycelia increased with increasing temperature. they also determined the optimal growth temperature in the range of 15°c–25°c and found no differences between 25°c and 30°c. the last observation was similar to that observed for our tested isolates. our group of “cold-grown” isolates refer to their lowest growth temperature (15°c), however, they were seen to grow the most intensively at 10°c. the growth of r. solani depends not only on temperature but also on the natural features of the isolates; the medium or soil type is also very important. r. solani is commonly known as a fast growing species. the importance of preferred growth temperature for r. solani showed by minier and hanson (2021) reported that below 15°c, no risk of rhizoctonia damping-off was observed on sugar beets. po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 20 other factors that may influence the r. solani growth rate in agricultural soils are fungicides. in this study, the following groups were investigated: strobilurin derivative — azoxystrobin, dithiocarbamate — thiuram, and benzimidazole — thiophanate-methyl. these compounds were found to inhibit the growth of r. solani mycelia, with thiophanate-methyl showing the highest antifungal activity. thiophanate-methyl at a dose of 100 ppm contributed to almost total growth inhibition in many of the tested isolates. two concentrations of fungicides were investigated in the study: lower (10 ppm) as an appropriate dose adequate for field applications to the soil, and higher (100 ppm), which was expected to completely inhibit the growth of fungi. kucharska et al. (2018) tested such compounds as propiconazole + cyproconazole, penthiopyrad, prochloraz, tebuconazole, and azoxystrobin against r. solani ag2-2b and ag4. all of these were able to notable reduce or inhibit the growth of r. solani. the least effective fungicide was azoxystrobin (efficacy of 47.30%–91.14% inhibition), whereas the most effective compounds were propiconazole and cyproconazole. moliszewska et al. (2020) showed that hymexazole, which is specific to oomycetes, could increase the growth of both tested fungi from 13.5% to 28%. however, thiuram inhibited them by 35.8%– 74.7% depending on the dose of the preparation. they concluded that both tested strains differed in response to both fungicides, which is similar to our observations for ag5 isolates. the differences observed for our isolates may be a result of the individual features inherited from the maternal isolate id23. this also shows that some level of potential resistance to fungicides could be acquired in the field by the maternal isolate or by natural genetic changes that may occur during long-term storage. our research shows that r. solani ag5 is represented by isolates differing in morphological features as well as some physiological and genetic properties. a more detailed study of this group is currently needed because r. solani ag5 has been isolated from different hosts. isolates have been found among other ags in soil, cucumbers, potatoes, tomatoes, peppers, red cabbage, wheat, turfgrasses, cereals, beans, soybeans, sugar beets, pine seedlings, and apple plants (ajayioyetunde and bradley ,2017; amaradasa et al. 2013; bełka and mańka, 2017, el-sharouny 2015; erper et al., 2021, fiers et al.,2011; melzer et al., 2016; shazad gondal and rauf, 2017; woodhall et al., 2012; yildrim and erper, 2017). recently, hassan and chang (2021) described the first case of damping-off in ovate-leaf atractylodes ovata caused by ag5. they may exist as single pathogens, as in the japanese monkshood (aconitum japonicum subsp. subcuneatum) (mori et al.,2020), wheat in the united kingdom or turkey, as well as barley and potato, or in mixed groups of different rhizoctonia-complexes in which they usually play a minor role (erper et al.,2011; gonzalez et al.,2001; moliszewska, po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 21 2009; woodhall et al., 2012; yildirim and erper, 2017). according to the results of moliszewska (1999), r. solani ag5 may comprise approximately 60% of the total amount of r. solani strains isolated from sugar beet seedlings in poland (moliszewska, 2009). r. solani ag5 can cause seedling damping-off and root rot in sugar beets and decrease plant size by slowing and stunting development (moliszewska, 2002); however, these observations require a more detailed inspection. recently, similar results were reported by erper et al. (2021) for red cabbage. some data suggest that members of ag5 may also act as severe pathogens, for example, on chickpeas in turkey (basbagci and dolar, 2021), hence, this strain seems to have become more dangerous than previously thought. they also should not be considered a homogenous group; as lübeck (2004) suggested, there are three possible groups based on rflp patterns. this was partially confirmed by the groups formed in the phylogenetic tree (figure 1). therefore, they should be studied to understand their evolution in nature and how they use plants as nutrient sources. knowledge gained from this type of research will allow the prevention of their potentially negative influence on crops in the future. conclusions research has shown that sexual reproduction leads to the segregation of features among progenies of the maternal isolate. in the tested isolates, several types of morphological features were observed, which were divided into two groups, one of the heterokaryons and the second of the homokaryons derived from basidiospores grouped together with their maternal isolate id23. this study is the first to show that sbi progenies mostly carry a morphology similar to the maternal isolate, but after crossing them, the next generation of heterokaryotic progenies creates a separate group of morphological characteristics (fig. 1). other attributes of new generations, such as the optimal temperature of growth or sensitivity to fungicides, also provide distinct information. a group of cool-preferring isolates that were sensitive to some fungicides were identified. higher sensitivity to fungicides has also been observed in heterokaryons than in mono/homokaryons. this study shows the changes in the next generation of r. solani. our observations of changes in progenies may contribute to our understanding of how r. solani ag5 modifies its aggressiveness to plants and resistance to fungicides. in our opinion, further research is needed to obtain fully described changes in the group ag5, to understand the details of its sexual reproduction, and to evaluate its notable in crops. po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 22 references abbott, w. s. 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(2012). first report of rhizoctonia solani anastomosis group 5 (ag5) in wheat in the uk. new disease reports, 26(1), 9–9. https://doi.org/10.5197/j.2044-0588.2012.026.009 po st pr in t vol. 57, article 578 doi: 10.5586/am.578 acta mycologica: postprint version 25 https://doi.org/10.17221/10480-pps https://doi.org/10.1016/j.sjbs.2020.05.026 https://doi.org/10.1007/s10327-020-00906-6 https://doi.org/10.1007/s10658-018-1447-2 https://doi.org/10.1080/15572536.2006.11832644 https://doi.org/10.1094/pdis-91-3-0260 https://doi.org/10.5197/j.2044-0588.2012.026.009 microfungi of the tatra mountains. part 7. correction of some data from herbaria and the literature 1 of 17published by polish botanical society acta mycologica review microfungi of the tatra mountains. part 7. correction of some data from herbaria and the literature monika kozłowska1*, wiesław mułenko1, kamila bacigálová2, agata wołczańska1, urszula świderska-burek1, magdalena pluta1 1 department of botany and mycology, maria curie-skłodowska university, akademicka 19, 20-033 lublin, poland 2 institute of botany, slovak academy of sciences, dúbravská cesta 14, 845 23 bratislava, slovakia * corresponding author. email: monika@poczta.umcs.lublin.pl abstract the tatra mts are located on the border of two countries – poland and slovakia. it is a unique, extremely geobotanically-differentiated region, protected by law and listed on the unesco biosphere reserve list as an internationally recognized area. due to the high nature values of the tatra mts, varied research, including mycological, has been intensively conducted on this area for many years. the first data on the microscopic fungi of the tatras comes from to the second half of the nineteenth century and spans more than 150 years. currently, the critical list of microfungi is being prepared concerning species published up to date from the whole tatra range (the polish and slovakian parts), and also the adjacent areas. during detailed study of the available mycological literature, many erroneous citations of the original data or incorrect interpretations of these records were noted. often, this faulty data was also reproduced in subsequent publications. the aim of this study was to correct some of the data published in the cited literature. in the paper, 68 fungal species were mentioned, including 29 species of ascomycota and 39 species of basidiomycota. additionally, some information about the plants – the fungal hosts – has also been corrected. keywords fungi; checklist; distribution; western carpathians, tatra national park; poland; slovakia introduction despite their relatively small surface area, the tatra mts are an important massif in central europe belonging to the western carpathians. they are the highest mountains between the alps and the caucasus mts and between the balkan peninsula and scandinavian mountains. the tatra mts are also the highest carpathian range crossing the migration path of arctic and alpine species between the east and west and between the north and south. although the tatra mts are significantly smaller and lower than the alps, the subnival belt develops in the tatras, making it the only place of such a type in central europe [1–3]. due to the specific and unique character of the flora and vegetation two national parks were established on the territory of the tatra mts: one in the slovakian tatras (tanap, tatranský národný park, 1949), and the second in poland (tpn, tatrzański park narodowy, 1954). the tatras are an area of interest for researchers representing various scientific disciplines, both in their range of animate and inanimate forms of nature. from the mycological point of view, the tatras flora and vegetation are of special value. the high richness of plants (about 1400 species) and their characteristic distribution, the doi: 10.5586/am.1081 publication history received: 2016-09-19 accepted: 2016-12-01 published: 2016-12-30 handling editor maria rudawska, institute of dendrology, polish academy of sciences, poland authors’ contributions mk, wm: contributed to the idea of research; all authors contributed to manuscript preparation; mk, wm: writing the manuscript; wm approved the final version of the manuscript funding the study was supported by the polish state committee for scientific research (grant no. 2/p04c/089/27 and no. n/ n304/172436), by the grant agency vega bratislava slovakia (projects no. 2/0106/10 and 2/0051/13) and from the department of botany and mycology, maria curieskłodowska university in lublin through its statutory funds. competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation kozłowska m, mułenko w, bacigálová k, wołczańska a, świderska-burek u, pluta m. microfungi of the tatra mountains. part 7. correction of some data from herbaria and the literature. acta mycol. 2016;51(2):1081. http://dx.doi. org/10.5586/am.1081 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:monika%40poczta.umcs.lublin.pl?subject=microfungi%20of%20the%20tatra%20mountains.%20part%207.%20correction%20of%20some%20data%20from%20herbaria%20and%20the%20literature http://dx.doi.org/10.5586/am.1081 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1081 http://dx.doi.org/10.5586/am.1081 2 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 occurrence of relict and endemic species, the differentiation of the geological substrate and the infinite number of terrain forms make the nature of the tatra mts particularly noteworthy [1–3]. these features are directly connected with a high richness of fungal species, including microfungi. mycological investigations have been intensively carried out on both sides of the tatra range, polish and slovakian. the first data about microscopic fungi was published in the second half of the nineteenth century by hazslinszky [4–7], kalchbrenner [8,9], and greschik [10,11]. they collected fungi on the south, slovakian side of the tatra mts. soon, polish researchers, e.g., krupa [12,13], raciborski [14–16], rehman [17], and boberski [18] also began studies, often on both sides of the tatras. the first collective summary of parasitic fungi occurring in the whole tatra range was prepared by stamachowa [19] in a hundred years since the beginning of the first studies. for the entire area of the tatra mts and adjacent areas, only 450 species of microscopic fungi were reported during that time. this number was underreported and incomplete; however, for the next 30 years, this work was cited as the main source of information. at the end of the twentieth century, several successive mycological works summarizing previous data were published. they concerned the zakopane basin (267 species) [20], the polish side of the national park (369 species) [21], and the slovakian side of the national park (283 species) [22]. however, at the beginning of the twentyfirst century, the first critical list of microfungi known from only the polish side of the park was published (461 species) [23]. a rapid increase in the number of known fungal species from this region was noticeable (100 species in 8 years), and every subsequent synthesis suggested that the richness of fungal species found in the tatra mts is much higher than hitherto recorded. in 1998, preliminary joint polish–slovakian mycological studies throughout the whole of the tatra mountains began. in the last decade (2005–2015), they were very intensively and systematically conducted under several research grants obtained from polish and slovakian institutions. one of the main goals of these projects, besides scientific research, was to prepare a complete, critical list of known microfungi in the whole tatra range and in its surroundings [24]. currently, the list of microfungi includes over 1700 species, while the cited mycological literature is comprised of more than 400 published scientific papers. aim and range of the study as mentioned earlier, the literature on the subject is rich, and simultaneously very diverse. it contains not only the original results of field studies, but also different types of syntheses, reviews, and popular science papers – in them, previous results have been cited, generalized, or transformed. during the study of all available literature about tatras fungi, the authors noted many erroneous citations or incorrect interpretations of the source data, which were later duplicated in subsequent publications. the causes of these inaccuracies can be conceived in a few points: ■ incorrect localization of localities; ■ incorrect identification of fungi and their hosts (mostly vascular plants); ■ incorrect citations of data from the original source papers; ■ omission of critical taxonomic studies, in which the significant proofs of existing data were made; ■ incorrect citations of papers (authors’ names, dates, titles, etc.) and random technical errors, uncritically treated as credible information. the objective of the present study is to show and correct some of the erroneous data about the microfungi of the tatra mts published in various papers. here, only the information that was cited incorrectly was included, and the introduced corrections did leave any doubt of the authors. in spite of these limitations, the list of reported mistakes is significant. some of our suggestions also apply to the need for a re-identification of herbarium collections (plants and fungi). modern methods of taxonomy have facilitated the separation of new species from existing ones (e.g., from genera 3 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 caltha, senecio, aconitum, and others). other, questionable data will be discussed in the aforementioned list of fungi being prepared for publication. some groups of fungi have already been the subject of our critical revision, and noted discrepancies have been corrected. this work concerns fungi belonging to two orders – taphrinales (ascomycota, including taphrinaceae and protomycetaceae) [25] and peronosporales (oomycota) [26], including members of the plasmopara genus published previously [27]. species belonging to the other groups, i.e., ascomycetous fungi (ascomycota; 29 species), and basidiomycetous fungi (basidiomycota; 39 species) have also been taken into consideration. moreover, the authors have revised some doubtful but available herbarium collections and corrected erroneous information about many host species of fungi. in keeping with the resolution that the present article is published in a continuous and generally accessible journal, our corrections will be readily available. we hope that this will contribute to significantly reducing the number of erroneous citations in the future publications concerning the fungi of the tatra mts. factors affecting the occurrence of erroneous information over the 150 years of mycological research, changes have taken place not only in the political sphere (two world wars, the collapse of austria-hungary, the revival of poland, and the dissolution of czechoslovakia) but also in geobotanical conditions. many mistakes may have been made due to the very strong internal differentiation and physiographic division of the tatra mountains as well as the mistranslation of old names, particularly those from hungarian and german. the research objectives and methods, the interpretation of results, and the means of publication have changed as well. the first botanical-mycological investigations carried out in this area had a typically preliminary nature, while at the same time providing scanty information about habitats and localities. with time, research became more detailed and took into account a greater number of habitat conditions. therefore, the literature offers highly diverse information ranging from simple lists of species identified in sporadic and occasional research to complex environmental analyses. the first explorers, e.g., hazslinszky, kalchbrenner, boberski [4,9,18] regarded the area of the tatra mountains as unified and called the region the “high tatras” (tatry wysokie, vysoké tatry, hohe tatra, magas-tátra). these were contrasted with the “low tatras” (nízke tatry, alacsony-tátra, niedere tatra), which are an isolated massif located approx. 30 km to the southwest. the first information about the region (both mycological and botanical) did not reflect the internal divisions of the tatra mountains. although the exact location of the localities is not specified, it is obvious that the aforementioned researchers worked on the southern, i.e., slovakian, part of the tatra mountains. others, e.g., krupa [12,13] and raciborski [14], carried out their observations and collections on the northern side, which currently belongs to poland. however, some polish researchers reported information from both parts of the tatra mts, and slovakian researchers have published data about fungi collected in the polish part of the region. a critical and cautious approach should be applied to the information published later by namysłowski [28,29], who sometimes changed or generalized the position of localities while citing other authors’ reports, e.g., krupa [13] and rouppert [30]. for instance, the information about the location of fungi in mountains near zakopane and kościelisko was reported stricte as zakopane town and kościelisko village. conversely, other authors published data about fungi collected in the town of zakopane and its surroundings with reference to those occurring in the mountains, thus covering the entire region as the tatra mountains (tatra national park). there were also serious difficulties in distinguishing between localities in areas with extremely similar names situated in different parts (eastern or western) of the tatra mountains and on both (polish and slovakian) sides of the massif. for instance, localities situated within the kościelisko village were often wrongly reported as “dolina kościeliska” valley. in the area of the tatra mountains, the name “świstówka” (slov.: svišťovka) appears repeatedly (pl – świstówka roztocka, świstówka waksmundzka, 4 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 mała świstówka, świstowa czuba; sk – svišťovka, malá svišťovka, veľká svišťovka, svišťovská dolina). furthermore, there are two areas called “tichá dolina” valley (tichá dolina liptovská and tichá dolina oravská), two so-called “five lakes valleys” (pl – dolina pięciu stawów polskich valley; sk – kotlina piatich spišských plies valley), several lakes called “black lake”, and many other examples. since there were no critical analyses of the original papers, the localities were incorrectly assigned to regions or countries. some localities reported from the tatra mountains refer to quite different regions. for example, kriváň mt, i.e., one of the highest peaks of the high tatras, was mistaken for fatranský (veľký) kriváň mt, which is the highest peak in the malá fatra massif (over a 50-km distance). five fungal species were wrongly reported as coming from the tatra mountains, although their localities were situated as far as in the ukraine. four of them referred to the name “volovec” (a peak in the western tatra mountains and a village in the ukraine) and one to kościelniki village (a village in the buchach district of the ukraine; orig. reported by raciborski) [14], which was mistaken for kościelisko village near zakopane by starmachowa [19]. two other species were reported as coming from the tatra mts by guyot [31], although they were collected in the surrounding area of kronstadt town (actually brașov) located in romania [32]. finally, the authors of the current study themselves have also not avoided erroneous citations, especially in papers published previously, before the preparation of this current, critical list of tatras fungi. list of the species1 ascomycota blumeria graminis (dc.) speer (erysiphales) ■ on dactylis glomerata l. on this plant, the fungus was reported from only two localities on the polish side of the tatra mts. it was also incorrectly reported from the vysoké tatry mts (sk) by bacigálowa [22] after paulech [33]. however, the latter author did not report this fungus on dactylis glomerata from the territory of the tatra mts. blumeria graminis is a commonly occurring fungus in the tatra mts and it was additionally recorded on 15 other plant species in this region. dangeardiella macrospora (j. schröt.) sacc. & p. syd. (dothideales) syn.: monographus macrosporus j. schröt. ■ on anthyllis alpestris (kit.) rchb. (fabaceae). on this plant, the fungus was incorrectly reported by starmachowa [19] after moesz [34]. however, in the original paper by moesz [34], the fungus was noted on “petioles of athyrii alpestris” [= athyrium alpestre (hoppe) nyl.] which belongs to polypodiales (ferns) and is a synonym of athyrium distentifolium tausch ex opiz. the fungus was collected by moesz [34] at malá studená dolina (sk, vysoké tatry mts). this region is located on granite rocks on which anthyllis alpestris does not occur; however, athyrium distentifolium is a common species. diatrype stigma (hoffm.) fr. (xylariales) ■ on fagus sylvatica l. in the paper by starmachowa [19], the fungus was reported from wołowiec mt (sk; the slovakian name – volovec mt, zapadné tatry mts, height 2064 m a.s.l.) after klika [35]. however, the record originally reported by the last author referred to volovec village located in western ukraine. at the 1 annotation: pl – poland, including the polish side of the tatra mts; sk – slovakia, including the slovakian side of the tatra mts; lbl – herbarium of maria curie-skłodowska university of lublin; sav – herbarium of the institute of botany at slovak academy of sciences in bratislava (both acronyms after index herbariorum: http://sciweb.nybg.org/science2/indexherbariorum.asp); m a.s.l. – meters above sea level; syn. – synonym; the bolded name of a fungus or plant (in various combinations) – species occurring in the tatra mts (sometimes on plants other than those listed); unbolded name of a fungus – species erroneously assigned as present in the tatra mts; unbolded name of a plant – species erroneously assigned as a host of fungus in the tatra mts; dolina – valley; kotlina – basin; pleso – lake; sedlo – pass. http://sciweb.nybg.org/science2/indexherbariorum.asp 5 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 beginning of the last century, the region was known as carpathian ruthenia (ruś podkarpacka, podkarpatská rus), and it belonged to czechoslovakia. also, the host plant does not occur at this elevation. notes. the same remark also applies to the other three species listed below. erysiphe heraclei dc. (erysiphales) syn.: erysiphe polygoni auct. p.p., erysiphe umbelliferarum de bary ■ on laserpitium latifolium l. on this plant, the fungus was earlier noted without localization by paulech et al. [36], and later in the monograph by paulech [33] reported from three localities from the slovakian side of the tatra mts: starý smokovec village (vysoké tatry mts), juráňova dolina (západné tatry mts), and oravice village (orava region). the information was cited by bacigálová [22]. however, during the recent revision of the herbarium collection in sav (bratislava) by k. bacigálová, the fungus was found only on the plant collected from the first locality. erysiphe polygoni dc. (erysiphales) ■ on phyteuma spicatum l. on this plant, the fungus was reported from the havran mt (belianské tatry mts, sk) by klika [35]. however, in the monograph by paulech [33] the fungus was not mentioned on this plant. also in the monographs by braun [37] and braun and cook [38], only leveilula taurica (lév.) g. arnaud is reported on the host. there is also no information about the host-fungus combination in fungal databases usda (http://nt.ars-grin.gov/fungaldatabases/). in the tatra mts, the fungus occurs on some other plant species (genera polygonum and rumex). erysiphe trifoliorum (wallr.) u. braun (erysiphales) syn.: erysiphe trifolii grev., microsphaera trifolii (grev.) u. braun ■ on trifolium arvense l. on this plant, the fungus was reported from the polish side of the tatra mts by three authors – namysłowski [29], starmachowa (as erysiphe martii lév.) [19], and sałata et al. [20] after the paper of krupa [13]. however, the last author did not report this host-fungus combination. in the tatra mts (pl and sk), the fungus is known to occur on another 14 species of plants. gibbera conferta (fr.) petr. (venturiales) ■ on vaccinium uliginosum l. on this plant, the fungus was reported by kubička [39] from trojrohé pleso at alt. 1600 m (sk, vysoké tatry mts, dolina bielych plies). the plant is probably vaccinium gaultherioides bigelow, the only species occurring at higher altitudes in the tatra mts. in earlier botanical literature, this species was not recognized. gibbera myrtilli (cooke) petr. (venturiales) syn.: venturia myrtilli cooke ■ on vaccinium uliginosum l. on this plant, the fungus was reported by picbauer [40] from the sedlo červenej hliny at alt. ca. 1400 m a.s.l. (sk, belianské tatry mts), and later cited by starmachowa [19]. the plant is probably vaccinium gaultherioides bigelow. at that height and in grassy places it is the only species occurring in the region (see also notes on gibbera conferta above). golovinomyces echinopis (u. braun) v. p. heluta (erysiphales) syn.: erysiphe echinopis u. braun ■ on stachys sylvatica l. on this plant, the fungus was reported from the vysoké tatry mts and belianské tatry mts (sk) by bacigálová [22]. however, the fungus belongs to neoërysiphe galeopsidis (dc.) u. braun, and is known on this plant from 10 localities, while golovinomyces echinopis occurs on echinops ritro l. and was reported from tatranská lomnica town (sk). hymenoscyphus callorioides (rehm) lizoň (helotiales) syn.: helotium callorioides rehm http://nt.ars-grin.gov/fungaldatabases/ 6 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 ■ on aconitum variegatum l. on this plant, the fungus was incorrectly noted by starmachowa [19] after svrček [41]. however, the latter author did not report the species on this plant but on dentaria bulbifera l. (belianské tatry mts, sk). hymenoscyphus vitellinus (rehm) kuntze (helotiales) syn.: helotium geiphilum velen. ■ on geum rivale l. the fungus was reported by velenovský [42] from the vysoké tatry mts (sk, environs of tatranska lomnicá town at alt. 1800 m a.s.l.) [orig.: m. tatra (1800) supra lomnice, 1924, leg. pilát]. later, the data was cited by svrček [43] and lizoň and jančovičová [44]. however, lizoň and jančovičová [44] suggested that the place of collection was the town of tatranská lomnica (ca. 800–900 m a.s.l.). we agree with this suggestion because the plant geum rivale does not occur at such high altitudes. lophodermium maculare (fr.) de not. (rhytismatales) ■ on vaccinium uliginosum l. on this plant, the fungus was reported by kubička [39] from trojrohé pleso at alt. 1600 m (sk, vysoké tatry mts, dolina bielych plies). however, the plant is probably vaccinium gaultherioides bigelow (see notes on gibbera conferta above). phaeosphaeria silenes-acaulis (de not.) l. holm (pleosporales) syn.: leptosphaeria silenes-acaulis de not. ■ on silene acaulis (l.) jacq. on this plant, the fungus was reported from five localities located in the slovakian part of the tatra mts. but, it was also incorrectly reported from the polish side of the region by sałata and mułenko [21] and mułenko et al. [23]. phyllachora heraclei (fr.) fuckel (phyllachorales) ■ on heracleum sp. on this plant, the fungus was reported by wróblewski [45] from dolina kościeliska (pl). however, in two later papers of starmachowa [19] and sałata et al. [20], the locality was incorrectly noted after rouppert [30] from kościelisko village. in the paper by rouppert (l.c.), there is no information on this fungus. the fungus was also reported in the region on heracleum sphondylium l. by wróblewski [46]. plowrightia noxia (ruhland) sacc. & d. sacc. (dothideales) ■ on the bark of fagus sylvatica l. in the paper by stamachowa [19], the fungus was reported from wołowiec (volovec) mt after klika [35]. however, the record should refer to volovec village located in western ukraine (see note on diatrype stigma above). podosphaera alpina (s. blumer) u. braun & s. takam. (erysiphales) syn.: sphaerotheca alpina s. blumer ■ on saxifraga rotundifolia l. on this plant, the fungus was reported from the tatra mts without precise localization by paulech et al. [36]. however, occurrence of the fungus in the tatra mts is doubtful, because the herbarium specimens were not preserved, and in the monograph by paulech [33], this information was not repeated. on this plant, the fungus was reported from the malá fatra mts and západné beskydy mts (sk). in the monograph of braun and cook [38], the information was generally cited from slovakia. podosphaera drabae (juel) u. braun & s. takam. (erysiphales) syn.: sphaerotheca drabae juel, sphaerotheca fuliginea auct. p.p. ■ on arabis alpina l. the fungus is known to occur in some localities located on the polish side of the tatra mts. however, the one polish record first reported by starmachowa [19] from wielka świstówka mt (pl; tatry wysokie mts, dolina rozto ki) was also incorrectly noted from slovakian side of tatry mts (belianské tatry mts) by paulech et al. [36], paulech [33], and bacigálová [22]. 7 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 podosphaera epilobii (wallr.) u. braun & s. takam. (erysiphales) syn.: sphaerotheca epilobii (wallr.) sacc. ■ on epilobium alpestre (jacq.) krock. on this plant the fungus was reported from dolina miętusia (pl) by starmachowa [19] and later cited by mułenko et al. [23], although in the earlier monograph by sałata [47] it was mentioned, that the fungus was incorrectly noted on this plant from the tatry mts. also bacigálová [22] wrongly cited the data reported by paulech [33] on epilobium alpestre; the latter author did not mention this host. on the territory of the tatra mts the fungus was collected on five other species of epilobium genus, from both parts of the region (pl and sk). podosphaera fuliginea (schltdl. ex fr.) braun & s. takam. (erysiphales) syn.: sphaerotheca fuliginea (schltdl. ex fr.) pollacci ■ on veronica sp. on this plant, the fungus was incorrectly noted by bacigálová [22] from the belianské tatry mts (sk) after paulech [33]. however, paulech (l.c.) reported it from the belianská dolina in the malá fatra mts (massif located approx. 50 km west of tatra mountains). in the territory of the tatra mts, the fungus is known to occur only on veronica chamaedrys l. podosphaera fusca (fr.) u. braun & shishkoff (erysiphales) syn.: sphaerotheca fusca (fr.) s. blumer ■ on doronicum austriacum jacq. on this plant, the fungus is known to occur in some localities on both sides of the tatra mts. however, on the same plant it was also noted by starmachowa [19] and paulech [33] after moesz [34] from päť spišských plies valley (sk, vysoké tatry mts). then, the record was quoted by bacigálová [22]. however, moesz [34] reported this fungus on doronicum stiriacum (vill.) dalla-tore, which is currently a synonym of doronicum clusii (all.) tausch. it was probably misinterpreted. the valley is located at an altitude above 2000 m. a.s.l. where doronicum austriacum occurs sporadically (or is not present), while doronicum clusii is a common species. podosphaera volkartii (s. blumer) u. braun & s. takam. (erysiphales) syn.: sphaerotheca volkartii s. blumer ■ on trifolium medium agg. this host/fungus combination was incorrectly noted by bacigálová [22]. the fungus is parasite of dryas octopetala l., and known to occur in a few localities in the region (pl and sk). podosphaera xanthii (castagne) u. braun & shishkoff syn.: sphaerotheca fusca (fr.) s. blumer ■ on petasites kablikianus tausch ex bercht. the fungus was wrongly noted on this plant in the monograph of the erysiphales of slovakia published by paulech [33] and later cited by bacigálová [22]. the host-fungus combination was reported from one locality (sk, vysoké tatry mts, between tatranská polianka village and sliezsky dom mountain hotel). the herbarian specimen was reidentified by w. mułenko. the host is adenostyles alliariae (gouan) a. kern. pyrenopeziza atrata (pers.) fuckel (helotiales) syn.: mollisia atrata (pers.) p. karst. ■ on senecio cordatus w. d. j. koch. on this plant, the fungus was reported by klika [35] from havran mt (sk, belianské tatry mts). the record is doubtful and collection needs revision. the plant species does not occur in this region (it is an alpine-apennine species). from the tatra mts the fungus was also reported on doronicum austriacum jacq. pyrenopeziza rubi (fr.) rehm (helotiales) ■ on gentiana asclepiadea l. on this plant, the fungus was reported by kubička [48] from the belianské tatry mts (sk). the collection needs revision because up to the time this fungus was not noted on the host. it is known to occur only on rubus idaeus l., also at the region. 8 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 rhytisma salicinum (pers.) fr. (rhytismatales) ■ on salix retusa l. on this plant, the fungus is known to occur in some localities on both sides of the tatra mts. it was also reported by hrubý [49] from päť spišských plies lakes (sk, kotlina piatich spišských plies). this data was later incorrectly cited by starmachowa [19] as located on the polish side of the tatra mts (dolina pięciu stawów polskich), and cited later in the same way by mułenko et al. [23]. rhytisma umbonatum hoppe (rhytismatales) syn.: rhytisma amphigenum (wallr.) magnus, rhytisma symmetricum jul. müll. ■ on salix caprea l. in the paper by starmachowa [19], the fungus was reported from wołowiec (volovec) mt after klika [35], and in the same way cited later by bacigálová [22] and mułenko et al. [23]. however, the record should be refer to volovec village located in western ukraine (see note on diatrype stigma above). sawadaea bicornis (wallr. ex fr.) homma (erysiphales) ■ on acer pseudoplatanus l. on this plant, the fungus is known to occur in two localities on the slovakian side of the tatra mts: juráňova dolina and oravice village (západné tatry mts) [33]. however, it was also reported by bacigálová [22] from the territory of the vysoké tatry mts on the basis of herbarium collections (sav, nový smokovec village, sep. 6, 1989, leg. c. pulech). the collection was revised by w. mułenko and identified as sawadaea tulasnei (fuckel) homma on acer platanoides l. sphinctrina turbinata (pers.) de not. (mycocaliciales) ■ on pertusaria sp. (lichens). on this lichen, the fungus was reported by lojka [50] from the environs of ždiar village (sk, spišska magura mts), and later cited by rehman [17], boberski [18], lisická [51], and kyselová [52]. in the last paper (kyselová l.c.), the fungus was additionally (but incorrectly) reported also from the polish side of the tatra mts after alstrup and olech [53] and bielczyk [54]; however, this locality is situated in the beskidy mts (pl). tapesia vaccini velen. (helotiales) ■ on vaccinium uliginosum l. on this plant, the fungus was reported by kubička [39] from dolina bielych plies (the environs of trojrohé pleso at alt. 1600 m, sk). however, only vaccinium gaultherioides bigelow occurs at this altitude (see also notes on gibbera conferta above). basidiomycota aecidium euphorbiae j. f. gmel. ex pers. (pucciniales) ■ on euphorbia cyparissias l. [tithymalus cyparissias (l.) scop.]. on this plant, the fungus was incorrectly cited by bacigálová [22] from the vysoké tatry mts (sk) after cejp and veselý [55]. however, the latter authors have reported this fungus from the surroundings of the town of trnava (near bratislava, the western part of slovakia). in the tatra mts, the fungus is known to occur on euphorbia sp. (kežmarok town) [56]. coleosporium tussilaginis (pers.) lév. (pucciniales) ■ on campanula pusilla haenke. on this plant, the fungus was noted by wróblewski [57] from mała dolinka pod giewontem (pl, tatry zachodnie mts). however, the herbarium collection was later revised by majewski [58] and identified as campanula serrata (kit.) hendrych (campanula napuligera schur). ■ on campanula scheuchzeri vill. all records on this plant were attached to campanula tatrae borbás on the basis of two critical revisions by majewski [58] and urban and marková [59]. on this host, the fungus is known to occur in seven localities from both sides of the tatra mts. ■ on phyteuma orbiculare l. on this plant, the fungus was reported from mały giewont mt (pl, tatry zachodnie mts) by wróblewski [57] and later cited by starmachowa [19] and mułenko et al. [23]. however, during an earlier revision of 9 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 herbarium specimens majewski [58] did not find the fungus on this host plant. it is therefore assumed that to date the fungus on this plant has not been collected. ■ on sonchus oleraceus l. on this plant, the fungus was reported from kościelisko village (pl, near the town of zakopane) by starmachowa [19] and sałata et al. [20] after wróblewski [45]. however, the latter author has reported the fungus from the dolina kościeliska (tatra national park). notes. coleosporium tussilaginis is a very common species; from the tatra mts, it was reported on 39 plant species. cronartium ribicola j. c. fisch. (pucciniales) syn.: cronartium ribicola a. dietr. ■ on pinus mugo turra. on this plant, the fungus was reported by hrubý [49] from four localities in the vysoké tatry mts (sk). however, the fungus occurs on pines possessing five needles on the brachyblast. according to urban and marková [59] it was evidently misidentified, and all records were attached to cronartium flaccidum (alb. & schwein.) g. winter. in the tatra mts, cronatium ribicola occurs on pinus cembra l., pinus strobus l. (cult.), and some species of ribes. endophyllum euphorbiae-silvaticae (dc.) g. winter (pucciniales) ■ on euphorbia amygdaloides l. [tithymalus amygdaloides (l.) hill]. on this plant, the fungus was reported from the belianské tatry mts (sk) by picbauer [60]. the record was later cited by majewski [58] and urban and marková [59]. however, starmachowa [19] and later bacigálová (after starmachowa) [22] wrongly cited this data after pilát [61]. the last author did not mention this species. hyalopsora aspidiotus (magnus) magnus (pucciniales) syn.: hyalopsora polypodii-dryopteridis (moug. & nestl.) magnus ■ on gymnocarpium dryopteris (l.) newman [phegopteris dryopteris (l.) fée] (ferns). in the tatra mts, the fungus is a common species known to occur on this plant on both sides of the region (pl and sk). however, it was also incorrectly reported from kriváň mt (the vysoké tatry mts, sk) by starmachowa [19] and bacigálová [22] after baudyš and picbauer [62]. however, in a recent publication, it was reported from the velky kriváň mt in the malá fatra mts, and later correctly reported by urban and marková [59]. melampsora caprearum thüm. (pucciniales) syn.: melampsora laricis-caprearum kleb. ■ on salix aurita l. on this plant, the fungus was incorrectly reported from slovakian side of the tatra mts by bacigálová [22] after pilát [61]; however, the latter author reported it as melampsora epitea thüm. (melampsora laricis-epitea kleb.). ■ on salix aurita l. × salix silesiaca willd. on this plant, the fungus was reported from the polish side of the tatra mts by starmachowa [19]; however, majewski [58] identified it as melampsora epitea thüm. melampsora euphorbiae (c. schub.) castagne (pucciniales) syn.: melampsora euphorbiae-dulcis g. h. otth., melampsora helioscopiae (pers.) g. winter ■ on euphorbia carniolica jacq. on this plant, the fungus was reported by wróblewski [63] from the polish side of the tatra mts (without localization), and later cited by starmachowa [19]. this collection is doubtful because according to majewski [58] the plant is not a component of mountain flora (it is a balkan species). in the region this fungus is found on two other plant species – euphorbia helioscopia l. and euphorbia platyphyllos l. melampsorella caryophyllacearum (dc.) j. schröt. (pucciniales) syn.: melampsora cerastii g. winter ■ on picea abies (l.) h. karst. on this plant, the fungus was reported by bacigálová [22] from the tomanovská dolina (sk, západné tatry mts). however, the 10 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 herbarium collection was revised by w. mułenko and identified as abies alba mill. on the latter plant, the species is known to occur in both sides of the region. microbotryum heliospermae piątek & m. lutz (microbotryales) ■ on heliosperma pusillum (waldst. & kit.) rchb. [heliosperma quadridentatum (murray) schinz & thell., ixoca pusilla (waldst. & kit) soják, silene pusilla waldst. & kit.]. on this plant, the fungus is known to occur in some localities on the polish and slovakian sides of the tatra mts. it was also cited by starmachowa [19] [as ustilago violacea (pers.) roussel] from “kopa under jaworzynka at alt. 1650 m” (sk) after součková [64]. however, součková (l.c.) did not report it from this locality. puccinia allii f. rudolphi (pucciniales) syn.: puccinia mixta fuckel, puccinia porri (sowerby) g. winter ■ on allium schoenoprasum l. subsp. sibiricum (l.) hay. on this plant, the fungus was reported from the belianské tatry mts (sk) by součková [65] and later cited by bacigálová [22]. however, the collection was revised by urban and marková [59] and the plant was identified as allium montanum f. w. schmidt [= allium senescens l. subsp. montanum (fr.) holub.]. puccinia arenariae (schumach.) g. winter (pucciniales) syn.: puccinia herniariae unger, puccinia spergulae dc. ■ on sagina procumbens l. on this plant, the fungus was wrongly cited by starmachowa [19] and later by sałata et al. [20] from kościelisko village near the town of zakopane (pl). however, in the original paper published in 1888 by raciborski [14], the correct location was kościelniki village (buchach district, the ukraine). in the territory of the tatra mts, the fungus is known to occur in many localities on some other plant species. puccinia calthae link (pucciniales) ■ on caltha laeta schott, nyman & kotschy [caltha palustris l. subsp. laeta (schott, nyman & kotschy) hegi] and caltha palustris l. (caltha palustris l. subsp. palustris). on these two plants, the fungus is known to occur in many localities and often noted on both plant species. however, these two species were not distinguished before. special attention should be paid to plants growing at higher altitudes where caltha laeta is the only species. for this reason, the whole collection should be revised. puccinia campanulae carmich. (pucciniales) ■ on campanula alpina jacq. on this plant, the fungus was reported by bacigálová [22] (after the herbarium of tanap, sk) and later repeated by urban and marková [59]. however, in this herbarium there is a collection of only campanula cochleariifolia lam. in the territory of the tatra mts, the fungus is known to occur on five other species belonging to the campanula genus. puccinia dentariae (alb. & schwein.) fuckel. (pucciniales) ■ on dentaria enneaphyllos l. on this plant, the fungus was incorrectly reported by starmachowa [19] and later by bacigálová [22] from the kriváň mt in the vysoké tatry mts (sk) after baudyš and picbauer [62]. however, in the latter paper, it was reported from a locality between the tatranský kriváň mt and terchova village. it was a misprint. the proper name is fatranský (veľký) kriváň mt in the malá fatra mts. the data was later properly reported by urban and marková [59]. puccinia dioicae magnus (pucciniales) ■ on carex brachystachys schrank. & moll. (carex tenuis host.). on this plant, the fungus was reported from the polish side of the tatra mts as puccinia sylvatica j. schröt. by raciborski [14] and namysłowski [28,29], and later cited by starmachowa [19] (as puccinia scabiosae-sempervirentis hasler). however, majewski [66] reidentified this species as puccinia caricina dc. these two species (puccinia dioicae and puccinia caricina) are very common in the tatra mts. 11 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 puccinia graminis pers. (pucciniales) syn.: puccinia anthoxanthi fuckel ■ on anthoxanthum odoratum l. s. l. on this plant, the fungus was reported by wróblewski from the town of zakopane [67] and dolina kościeliska [57] (pl), and later cited by starmachowa [19], in all papers as puccinia anthoxanthi fuckel. during majewski’s revision [66], the fungus was identified as puccinia poae-nemoralis g. h. otth. on the same plant, the fungus was also incorrectly cited by bacigálová [22] after picbauer [40] from the slovakian side of the tatra mts; however, the latter author did not report it in the paper. ■ on deschampsia flexuosa (l.) trin. (aira flexuosa l.). on this plant, the fungus was noted by wróblewski [57] from dolina kościeliska (pl) and from temnosmrečinská dolina (sk), and later cited by starmachowa [19] and bacigálová [22]. according to majewski [66], the record from the polish side of the region is uromyces airae-flexuosae berd. & winge and the record from the slovakian side is puccinia deschampsiae arthur on deschampsia caespitosa (l.) p. beauv. however, in the monograph by urban and marková [59], this record was omitted. ■ on elymus caninus (l.) l. (triticum caninum l.). on this plant, the fungus was reported by wodziczko [68] from dolina kościeliska (pl, leg. żmuda, aug. 1910); however, in the papers by namysłowski [28,29] and starmachowa [19] the place was incorrectly reported as kościelisko village. notes. puccinia graminis is a common species in the tatras; it was reported on nine plant species. puccinia hieracii (röhl.) h. mart. (pucciniales) ■ on hieracium glaucum all. on this plant, the fungus was reported by krupa [13] (as puccinia flosculosorum auct. p.p.) from the polish side of the region and later cited by namysłowski [28,29]. the record is probably doubtful. in the monograph by majewski [66], the plant is not mentioned as a host of the fungus, and there is no information on this plant in the polish checklist of plants [69]. however, puccinia hieracii is a very common species in the region, noted on 21 other species of plants. puccinia morthieri körn. (pucciniales) ■ on geranium sylvaticum l. on this plant, the fungus was reported by wróblewski [57] from mały giewont mt (pl, tatry zachodnie mts). this collection was revised by majewski [66], and the fungus was identified as puccinia leveillei mont. however, both species are known to occur in the region. puccinia obscura j. schröt. (pucciniales) syn.: puccinia luzulae-maximae dietel, puccinia obscura var. luzulae-maximae (dietel) u. braun ■ on luzula luzulina (vill.) dalla torre & sarnth. [luzula flavescens (host.) gaud.]. in the paper by kochman and sałata [70], the locality was incorrectly reported as the łysica mt in the świętokrzyskie mts (pl) (mycotheca polonica, fasc. xxvi– xxx, no. 715). however, the original collection is in the herbarium of lbl (sep. 27, 1978, leg. t. majewski), and the correct place is the town of zakopane (pl, kotlina zakopiańska). from this place, it was later correctly cited by sałata et al. [20]. puccinia pygmaea erikss. (pucciniales) ■ on calamagrostis varia (schrad.) host. on this plant, the fungus was reported by wróblewski [57] from dolina za bramką (pl, tatry zachodnie mts) and later cited by starmachowa [19]. according to majewski [66], it is puccinia striiformis westend. on elymus caninus (l.) l. [agropyron caninum (l.) p. beauv.]. however, puccinia pygmaea is known to occur on two other host plants from some localities on both sides of the tatra mts. puccinia ribis dc. (pucciniales) ■ on ribes alpinum l. on this plant, the fungus is reported from both sides of the tatra mts. the herbarium collections were revised by majewski [66] and urban 12 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 and marková [59], and the plant was identified as ribes petraeum wulfen. the fungus is known to occur in many localities in the region. puccinia scorzonerae (schumach.) jacky. (pucciniales) ■ on scorzonera purpurea l. on this plant, the fungus was reported from the polish side of the tatra mts without precise localization by wróblewski [63], and later cited by starmachowa [19]. according to majewski [66], the occurrence of this fungus in the tatra mts is uncertain. the plant does not occur in the region, and herbarium specimens were not preserved. puccinia soldanellae (dc.) fuckel (pucciniales) ■ on soldanella hungarica simonk. [soldanella montana vill. subsp. hungarica (simk) lüdi, soldanella major (neilr.) vierh.]. on this plant, the fungus is known to occur in some localities on both sides of the tatra mts (pl and sk). it was also reported from kościelisko village (near the town of zakopane, pl) by rouppert [30] and later cited by starmachowa [19] on soldanella alpina l. (= soldanella montana will.). however, rouppert’s collection (leg. k. rouppert, sep. 14, 1909, kościelisko village – slope of hruby regiel mt) was earlier published by namysłowski [28] (on soldanella alpina l.) who incorrectly reported this record from dolina kościeliska (tatra national park). puccinia striiformis westend. (pucciniales) syn.: puccinia glumarum (j. c. schmidt) erikss. & henning ■ on calamagrostis epigejos (l.) roth. on this plant, the fungus was reported by picbauer [40] (as puccinia glumarum) from the belianské tatry mts (sk), and cited in the same way by starmachowa [19] and bacigálová [22]. however, it was challenged by urban [71] and later reidentified as puccinia pygmaea erikss. var. pygmaea on calamagrostis sp. [59]. in this region, the puccinia striiformis is known to occur on elymus caninus (l.) l. puccinia swertiae (opiz) g. winter (pucciniales) ■ on swertia perennis l. the fungus is known to occur in many localities on both sides of the tatra mts. on this plant, it was also reported by wróblewski [57] from the surroundings of zmarzły staw (eng. frozen pond) lake in the tatry wysokie mts (pl). however, the same record was incorrectly reported by urban and marková [59] from the lake with a similar name (zmrzlé pleso), which is located on the slovakian side of the tatra mts. puccinia tanaceti dc. (pucciniales) syn.: puccinia pyrethri (wallr.) rabenh. ■ on dendranthema zawadzkii (herbich) tzvelev (chrysanthemum zawadzkii herbich). on this plant, the fungus was reported from the tatra mts by jørstad [72] (leg. ullepitsch, without localization). according to majewski [66], this record was noted erroneously because the plant does not occur in the tatra mts. it occurs mainly in asia, and in europe it is known only from the pieniny mts (pl and sk). however, puccinia tanaceti is known to occur in a number of localities on three other plant species from the genus artemisia and tanacetum. puccinia violae dc. (pucciniales) ■ on viola bicolor pursh. on this plant, the fungus was reported from dolina javorinka (sk, belianské tatry mts) by klika [35]. according to starmachowa [19] and urban and marková [59], it is a misprint and the plant name is viola biflora l. which is infected by puccinia alpina fuckel. however, puccinia violae is known to occur on some other species of the viola genus. puccinia virgae-aureae (dc.) lib. (pucciniales) ■ on solidago virgaurea l. on this plant, the fungus was reported without precise localization from the polish side of the tatra mts by wróblewski [57,63], and later cited by starmachowa [19] and mułenko et al. [23]. however, according to majewski [66] the record is uncertain and the herbarium collection was not preserved. 13 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 on the same plant species, the fungus was also collected on the slovakian side of the tatra mts. in this case, all data were included to solidago alpestris waldst. & kit. [solidago virgaurea l. subsp. alpestris (w. k.) gaud., solidago virgaurea l. subsp. minuta (l.) arcang.] on the basis of critical revisions of urban and marková [59]. it seems that the polish data should also be included in this species. trachyspora intrusa (grev.) arthur (pucciniales) syn.: trachyspora alchemillae (pers.) fuckel, uromyces alchemillae (pers.) fuckel ■ on alchemilla baltica sam. ex juz. on this plant, the fungus was incorrectly cited from the západné tatry mts (sk) by bacigálová [22] after urban [73]. however, urban [73] did not mention the plant in this publication. the fungus is known to occur on many other species of the alchemilla genus. uredinopsis filicina (niessl) magnus (pucciniales) ■ on gymnocarpium dryopteris (l.) newman [phegopteris dryopteris (l.) fée]. on this plant, the fungus was reported by starmachowa [19] and later cited by mułenko et al. [23]. however, majewski [58] reidentified the plant as phegopteris connectilis (michx.) watt (phegopteris polypodioides fée), on which the fungus is known to occur in numerous localities on both sides of the tatra mts (pl and sk). uredo ribicola lasch (pucciniales) ■ on ribes alpinum l. on this plant, the fungus was reported by kalchbrenner [9] from lučivná village near poprad town (sk). however, the collection was revised by urban and marková [59] and identified as puccinia ribis dc. on ribes petraeum wulfen. the last species (on the mentioned host) is common in the region. uromyces acetosae j. schröt. (pucciniales) ■ on rumex acetosella l. on this plant, the fungus was reported from polish side of the tatra mts by krupa [13] and later cited by namysłowski [28,29] and starmachowa [19] under two names, as uromyces acetosae j. schröt. and additionally as puccinia acetosae körn. however, it was revised by majewski [66] and identified as uromyces polygoni-aviculariae (pers.) p. karst. and cited later in the same way by sałata et al. [20]. ■ on rumex alpestris jacq. [acetosa arifolia (all.) schur., rumex arifolius all.]. on this plant, the fungus was reported from temnosmrečinská dolina and liptovské múry mt (sk, vysoké tatry mts) by urban [73]. this record was later twice quoted by starmachowa [19] – once correctly as uromyces acetosae and once incorrectly, as uromyces rumicis (schumach.) g. winter. uromyces airae-flexuosae ferd. & winge (pucciniales) ■ on deschampsia flexuosa (l.) trin. [aira flexuosa l., avenella flexuosa (l.) parl]. on this plant, the species is known to occur in some localities in the region. however, it was also noted from západné tatry mts (sk, between rákoň mt and roháč mt) by součková [74]. according to urban and marková [59], it is puccinia festucae plowr. on festuca picta kit. uromyces anthyllidis (grev.) j. schröt. (pucciniales) ■ on anthyllis polyphylla kit. ex dc. on this plant, the fungus was incorrectly reported from the tatra mts by guyot [31: p. 160] after magnus [32]. however, magnus [32] reported it from kronstadt town (= brașov town, romania). in the tatra mts, the fungus is known to occur on two other plants – anthyllis alpestris (kit.) rchb. and anthyllis vulneraria l., but all collections require revision. until recently, these two species were not distinguished, and only anthyllis vulneraria was given. uromyces cacaliae (dc.) unger (pucciniales) ■ on adenostyles alliariae (gouan) a. kern. (adenostyles albifrons reichenb.). on this plant, the fungus is known to occur in some localities in the region. however, it was also incorrectly cited by starmachowa [19] from malá studená dolina (sk, 14 of 17© the author(s) 2016 published by polish botanical society acta mycol 51(2):1081 kozłowska et al. / microfungi of the tatra mountains. part 7 vysoké tatry mts) after husz [75]. however, husz [75] did not provide this information. this record was published by hrubý [49]. uromyces geranii (dc.) fr. (pucciniales) ■ on geranium sylvaticum l. on this plant, the fungus is known to occur in some localities from both sides of the tatra mts (pl and sk). additionally, it was incorrectly reported from the tatra mts by guyot [76: p. 257] after magnus [32]. however, magnus [32] reported it from kronstadt town (= brașov town, romania). uromyces phyteumatum (dc.) unger (pucciniales) ■ on adenostyles alliariae (gouan) a. kern. (cacalia alpina mill.). on this plant, the fungus was reported by hazslinszky [5] from the slovakian side of the tatra mts (without localization). however, adenostyles alliariae can be infected by two uromyces species: uromyces veratri (dc.) j. schröt. and uromyces cacaliae (dc.) unger [66]. on the basis of the description of teliospores, the fungus was included to uromyces cacaliae, which was also reported by kalchbrenner [9]. in the tatra mts, the uromyces phyteumatum is known to occur only on phyteuma spicatum l. and was noted in two localities on the polish side of the region. uromyces pisi (dc.) g. h. otth (pucciniales) ■ on lathyrus pratensis l. on this plant, the fungus is known to occur in some localities in both parts of the tatra mts and surroundings. it was also reported by starmachowa [19] from the wołowiec (volovec) mt after klika [35]. however, the record should be refer to volovec village located in western ukraine (see also notes on diatrype stigma, ascomycota). uromyces rumicis (schumach.) g. winter (pucciniales) ■ on rumex acetosa l. on this plant, the fungus was reported by wróblewski [57] from the town of zakopane (pl). the herbarium collection was revised by majewski [59] and identified as puccinia acetosae körn. in the region, however, the uromyces rumicis is known to occur on some other plant species. acknowledgments special thanks to prof. tomasz majewski for his help in the taxonomical revision of the fungi, for providing many unique positions on the literature and on records, and for his help in determining a detailed distribution of many fungi in the tatra mts. references 1. mirek z, editor. przyroda tatrzańskiego parku narodowego. kraków: tatrzański park narodowy; 1996. 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(encyclopédie mycologique; vol 15). abstract introduction aim and range of the study factors affecting the occurrence of erroneous information list of the species ascomycota basidiomycota acknowledgments references 2016-12-30t12:27:07+0000 piotr otręba scientific research conducted at the department of mycology, university of warmia and mazury in olsztyn acta mycologica article id: 5525 doi: 10.5586/am.5525 publication history received: 2020-06-15 accepted: 2020-07-22 published: 2021-02-05 handling editor wojciech pusz; wrocław university of environmental and life sciences, poland; https://orcid.org/0000-00031531-2739 authors, contributions ab: concept for publication, chapter 2 preparation, manuscript correction, final manuscript revision; ee: chapter 3 preparation; dk: chapter 4 preparation, manuscript correction, final manuscript revision; es: chapter 5 preparation funding this publication was financed by the statutory funds of the department of microbiology and mycology, faculty of biology and biotechnology, university of warmia and mazury in olsztyn. competing interests no competing interests have been declared. copyright notice © the author(s) 2020. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. review in mycology scientific research conducted at the department of mycology, university of warmia and mazury in olsztyn anna biedunkiewicz *, elżbieta ejdys , dariusz kubiak , ewa sucharzewska department of microbiology and mycology, faculty of biology and biotechnology, university of warmia and mazury in olsztyn, poland *to whom correspondence should be addressed. email: alibi@uwm.edu.pl abstract this paper presents a review of the research conducted by the staff of the department of mycology at uwm, olsztyn since its establishment to the present. this unit was established and has been headed for over 20 years by prof. maria dynowska. since 2004, the department has been conducting extensive mycological research, which is reflected in the dynamic growth of specialist staff involved in teaching activities and popularizing scientific research. owing to the particular care of prof. dynowska, and maintenance of the principal interdisciplinary character of the research, the department has been occupying a significant position in mycology in poland recently. this paper attempts to provide a summary of the major scientific accomplishments of the team headed by prof. dynowska. keywords potentially pathogenic fungi; aquatic ecosystem; school environment; urban environment; lichenological studies; plant parasites; bioindication 1. introduction the department of mycology of the university of warmia and mazury in olsztyn was established on april 1, 2004. it was the product of the transformation of the department of mycology established in 1996 from the department of botany at the institute of biology and environmental protection, faculty of mathematics and life sciences of the teachers college in olsztyn. prof. dr. hab. maria dynowska was appointed head of the department in 1996. initially, the research conducted at the department covered the issues of botany and plant physiology in a broad sense. over time, the subject matter evolved toward pure mycology. in consequence, mycology was isolated from botany, and the teaching offer was expanded to include obligatory and elective mycological subjects (general and systematic mycology, lichenology, phytopathology, medical mycology, hydromycology, mycosociology, mycoindication, mycological diagnostics, applied mycology, etc.), whose status in the syllabus was adjusted to that of the major subjects (biology, biotechnology, microbiology, and nursing), and two research teams were appointed: the ecology and environment monitoring team and the applied mycology team. the teams included mycologists and botanists, who prepared an assessment of the environment against which fungi were analyzed (saprotrophs, phytopathogens, zoopathogens, anthropopathogens, and lichenized fungi). upon the establishment of the university of warmia and mazury in olsztyn, the scope of research was largely modified, and some of the staff of the department of mycology were transferred to other university units. owing to the initiative of prof. dynowska, new research teams, viz., the fungi and lichens monitoring team and the medicinal and applied mycology team, were appointed. these acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 1 https://doi.org/10.5586/am.5525 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0003-3498-7432 https://orcid.org/0000-0002-5313-9247 https://orcid.org/0000-0001-7266-0647 https://orcid.org/0000-0001-6754-5994 mailto:alibi@uwm.edu.pl biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) teams have operated throughout the history of the department. the research was interdisciplinary, and covered issues in mycology, microbiology, phytopathology, lichenology, and ecology, with emphasis on the diversity of fungi in macroand microenvironments and the significance of fungi in bioindication (in a broad sense of the term). it was conducted based on two multitask subjects: • biological and ecological factors that affect fungi diversity and expansiveness; • fungi and lichens against natural plant communities. by presenting the findings of the research conducted at the department via this review paper, we wish to express our gratitude to prof. dynowska for her scientific supervision, care for the development of our research, and continuous support for our cooperation with various research centers. each chapter was prepared by prof. dynowska’s protégés, and presents diverse specialties of mycology. 2. potentially pathogenic fungi isolated from waters and their application in bioindication research on the presence of fungi in various types of waters has been conducted around the world since the 1960s. it has dealt with mycological environmental monitoring in general terms (cooke, 1965; laundon, 1972), and resulted in the development of standards of cleanliness in the usa and canada, with fungi as bioindicators of pollution in household, industrial, agricultural, and touristic wastewater (american society for testing and materials, 2005). hydromycological research in poland has been conducted with regard to the number, species diversity, and ecophysiology of fungi in waters of different trophic states, with attention also being paid to potentially pathogenic species (dynowska, 1995; kurnatowski et al., 2007). continuous mycological monitoring of waters in north-east poland was initiated in the 1990s by prof. dynowska (dynowska, 1993b, 1993d, 1995, 1997), and it has been continued by her research team. the scope of research has been expanded to include water bodies in the north-west (biedunkiewicz et al., 2007) and central parts of poland (biedunkiewicz & ozimek, 2009; ozimek & biedunkiewicz, 2008). the findings of the research not only explain the role of microfungi in the trophic and energetic loops in water bodies, but also help in identifying specific fungi with indicative properties, which can be used in a multiaspect assessment of waters, especially regarding their sanitary and epidemiological quality (biedunkiewicz, 2011b; dynowska, 1995, 1997). the hydromycological research focused on microfungi (potential human pathogens) present in different types of waters, their numbers and taxonomic differentiation during all seasons of the year, and those with interesting epidemiological and bioindicative features. analyses were performed for tap and potable bottled water (biedunkiewicz et al., 2014; biedunkiewicz & schulz, 2012), and water from places used for recreation, including municipal lakes and indoor swimming pools (biedunkiewicz, 2007a; biedunkiewicz & góralska, 2016; biedunkiewicz et al., 2007), as well as fountains situated in parks and at squares used by residents for leisure (biedunkiewicz, 2009). it was shown that, in the immediate vicinity of bathing sites, the existing littoral zone retained yeasts (potential human pathogens) on the phyllosphere surface. long-term observations in one of the lakes in olsztyn have shown the presence of the same microfungi species in both habitats in 60% of cases. the presence of many potentially pathogenic yeast species in the lake littoral zone shows the huge impact of humans on the hydrosphere microbiological quality (biedunkiewicz et al., 2020). the frequent presence of microfungi (including candida albicans, c. tropicalis, c. guilliermondii, and c. krusei) in waters used, in a variety of ways, by humans for consumption or recreation has prompted observations of some ecophysiological features of these fungi, such as the multiplication rate, enzymatic activity, and pseudomycelium growth capability in controlled laboratory conditions, with the use of modified media based on filtered water acquired from the natural environment, and a photoperiod appropriate for the given environmental and daily temperature fluctuations. one of the parameters – enzymatic activity – was tested before and acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 2 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) after culturing four of the aforementioned fungi species in the laboratory. the results showed that the microfungi retained a high physiological activity. hence, they should be entered in the list of pathogens whose presence in utility waters, including thermal and recreational waters, disqualifies them from public use. one cannot exclude the role of water as a habitat for potential pathogens, because depending on the type, amount, and availability of organic matter, numerous fungi species find favorable trophic conditions for development, which have a direct impact on the metabolism rate (biedunkiewicz, 2015). the study was conducted as part of a research grant from the ministry of science/national centre for science, entitled “próba zastosowania grzybów potencjalnie chorobotwórczych dla człowieka w standardowych metodach oceny czystości wód powierzchniowych [an attempt to use fungi – potential human pathogens – in standard methods of surface water cleanliness evaluation].” as a permanent component of inland water microbiota, fungi were present in all the types of water under study. however, an increase in their number depended on the degree of anthropopressure and eutrophication. the eutrophic lakes were found to contain numerous, frequently repetitive species of potential pathogens or saprophytes; some fungi that had not been reported in the literature on hydromycology were also isolated, viz., candida kruisii and dipodascus armillariae (biedunkiewicz & baranowska, 2011). the research conducted at the department to date has demonstrated the diversity of microfungi in waters and the increase in their numbers in proportion to the pace and intensity of anthropogenic changes. this applies mainly to fungi that prefer strongly eutrophicated waters that are rich in organic matter of various origins. these fungi include ascomycetes and basidiomycetes, as well as some molds. it must be emphasized that the majority of the species of these microfungi groups isolated from waters are potential aetiological agents of surface and organ mycoses, and their abundant proliferation in the waters under study may pose a serious epidemic threat. owing to this, a relatively complete picture of the water quality can be shown with respect to the sanitary and epidemic status, taking into account fungi with high pathogenic potential (biedunkiewicz et al., 2013). the majority of fungi isolated from water are opportunistic, and can settle on the human skin and mucous membrane, most frequently in the respiratory and alimentary tracts (biedunkiewicz, 2011a). this may be temporary settlement or colonization, which does not produce symptoms in carriers, while people with impaired immunity acquire mycotic infection. the abundance of fungi that colonize various sections of the respiratory tract was demonstrated in a multiyear study conducted at samodzielny publiczny zespół gruźlicy i chorób płuc [independent public tuberculosis and pulmonary disease unit] in olsztyn, which was the starting point for the hydromycological studies described herein. these studies produced surprising findings and provided the basis for the doctoral thesis entitled dynamika mikoflory układu oddechowego człowieka [mycoflora dynamics in the human respiratory tract], with prof. dynowska as a scientific supervisor. the analyses revealed the presence of 29 species of eight genera, viz., candida, geotrichum, saccharomyces, saccharomycopsis, schizosaccharomyces, torulopsis, trichosporon, and aspergillus. many of these fungi have been isolated in the hospital environment, indicating that the same species present in the human environment can appear in this ontosphere under favorable conditions, i.e., immunosuppression and dysbacteriosis (biedunkiewicz, 2001; dynowska et al., 2002, 2013). these findings suggested the need for mycological analyses in healthy individuals, i.e., students of biology and veterinary medicine (biedunkiewicz, 2007b, 2011a). the results confirmed the carriage of 15 species of potentially pathogenic fungi of seven genera in nearly 80% of the tested individuals. combined with the results of clinical observations, they directed the researchers’ attention to natural reservoirs of fungi in immediate human surroundings. the list of fungi that have been isolated from waters to date is long, and contains 185 species. each aquatic ecosystem under study was found to contain c. albicans, although it was not always a dominant species. it is particularly important in the context of the relationship of the species with the ontospheres of humans and acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 3 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) other vertebrates, which poses a potential violation of the biological balance, and breakdown of body immunity. the presence of c. albicans in polluted waters suggested its usability as an indicator of cleanliness and sanitary security, especially since this species survives longer in marine and fresh waters than escherichia coli (biedunkiewicz, 2015; dynowska, 1995). microfungi also survive at low temperatures. the psychrosphere is not a habitat barrier for them. this has been confirmed via mycological analyses of ice and snow cornices (biedunkiewicz & ejdys, 2011; ejdys, biedunkiewicz, et al., 2014). all species proved to be psychrotolerant, and they retained high viability following incubation at 37 ◦c. these research findings provide grounds for regarding ice and snow cornices as natural and temporary air filters and specific vectors of potential pathogens, which pose a real epidemic threat in the environment. the hydrosphere is one of the greatest fungi reservoirs with a diverse taxonomic and trophic status. in addition to saprotrophs with dormant pathogenic potential, species regarded as human and animal pathogens have been isolated from water bodies. quantitative and qualitative assessments of isolated microfungi are very important in comprehensive mycological studies of various aquatic ecosystems and utility waters. an analysis of the prevalence of their occurrence in recreationally used waters, especially in summer, is imperative. there may be no officially published epidemiological data concerning the correlations between microfungi concentrations in waters and infections in humans who have contacts with them; however, the occurrence of fungi in waters suggests such a possibility, and also indicates that a water reservoir can be a serious source of mycoinfections. current knowledge of the ecophysiology of potentially pathogenic fungi and longterm studies indicate that waters are important links in the mycoses epidemic chain in the biosphere, especially if they contain fungi from household waste, asymptomatic carriers, or people with mycoses (dynowska, 1995). therefore, important achievements in hydromycological research include the development of a procedure for the isolation, counting, and identification of potentially pathogenic fungi, which can be used by laboratories in routine sanitary and epidemiological testing of waters used for various purposes. this was incorporated, together with six key publications (biedunkiewicz, 2011b, 2015; biedunkiewicz & baranowska, 2011; biedunkiewicz & ejdys, 2011; biedunkiewicz & góralska, 2016; biedunkiewicz & ozimek, 2009), in the scientific accomplishment entitled “możliwości wykorzystania wybranych mikrogrzybów do oceny sanitarno-epidemiologicznej wód na tle różnorodności i fenologii gatunków potencjalnie chorobotwórczych dla człowieka [use of selected microfungi in sanitary and epidemiological assessment of water against the background of diversity and phenology of potential human pathogens],” which was the basis for conferring the scientific degree of doctor habilitated in biological sciences, microbiology specialty. the research conducted to date at the department has been devoted to the taxonomy, quantitative aspects, and phenology of isolated fungi against the background of natural and anthropogenic factors in aquatic environments. the research findings have been published in nearly 110 scientific papers, showing a relatively complete picture of water quality with respect to its sanitary and epidemic status, taking into account fungi with high pathogenic potential (dynowska & biedunkiewicz, 2013). 3. circulation of potentially pathogenic fungi in a school environment the research conducted by prof. dynowska on potentially pathogenic fungi in the biosphere was expanded in the 2000s to include analyses of their occurrence in organ ontocenoses in healthy children and adolescents in rural and urban environments. the research conducted for the doctoral thesis entitled analiza mikologiczna wybranych ontocenoz dzieci w wieku 6–15 lat [mycological analysis of selected ontocenoses in children aged 6–15 years] (ejdys, 2002) was used as the starting point. it showed that schools were very interesting and poorly explored objects of mycological research, especially with respect to the factors affecting the occurrence of fungi in children and adolescents, as well as their surroundings. the acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 4 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) results also encouraged researchers to plan studies of importance to epidemiology, concerning multidirectional and multilevel analyses of the transmission of these fungi. the greatest attention in pediatric mycology literature is devoted to children in risk groups. they include children with diabetes and cancers undergoing intensive chemotherapy, and immunosuppression and antibiotic therapy, or those being treated orthodontically. few papers have been published on mycobiota in children and adolescents regarded as healthy. the first/pilot study was conducted in 1997 on 70 children aged 0–9 and 10–15 years (dynowska & ejdys, 2000), and was aimed at determining interindividual transmissibility within same-age groups. fortyfive isolates of eight fungi species were obtained. these species were present in 39% of the population under study, and they occurred in children more frequently than in adolescents. the phenological differences in the prevalence of fungi in the children under study were emphasized. unlike in adults (dynowska, 1993a), fungi were found twice more frequently in children’s ontocenoses in spring than in fall. obtaining multispecies isolates was an important outcome of this research. considering the findings and the epidemiological importance of the artificially created population of children who stay as a group within a limited area of a school building for up to 9 hours a day, it was decided to expand the school environment research. over three subsequent years, 270 generally healthy children aged 6– 15 years and living in the voivodship of warmia and mazury were examined (ejdys, 2001, 2003a, 2003b). the research aimed to evaluate the relationship between the occurrence of fungi in children’s oral cavities and noses, and the living environment (town–village), gender, age, diet, prior diseases, antibiotic therapy, and contacts with the hospital environment, as well as identify children at risk of mycotic infections. fungi were found in 36.3% of the population tested, which was a figure comparable to that in preliminary examinations; 168 isolates of 13 yeast species were obtained. candida albicans – the main aetiological agent in the majority of human mycoses – was a dominant species. the presence of c. glabrata and c. krusei, whose increasing number of strains demonstrated resistance to main antimycotics, and the relatively frequent occurrence of trichosporon beigelii, saccharomycopsis capsularis, and saccharomyces spp. – fungi of growing expansiveness and enzymatic activity – was a cause for concern. regardless of the season, the greatest fluctuations in the mycocenoses diversity and count were observed in the oral cavity and nose, which were the organs in direct contact with the contaminants in the environment and the many potentially pathogenic fungi transmission agents. the threat of a mycotic infection in the majority of the tested population increased after antibiotic therapy, regardless of the season of the year or the age. this mainly concerned the most stable ontocenosis of the pharynx. children living in villages, who had been ill immediately before the material was collected, were at the greatest risk of having fungi in any of the ontocenoses under study. this is a consequence of, among other things, more frequent contact of children in a rural environment with natural reservoirs rich in fungi (ejdys & dynowska, 2004). since the differences between the prevalence of fungi in girls and boys living in villages and those living in towns may have been caused by physiological, rather than environmental factors, a study aimed at determining the effect of gender on the frequency of fungal infections in children of school age was conducted (ejdys, 2008a). in spring and fall, tests were performed on 238 girls and 206 boys (regarded as healthy) of the following ages: 6–9, 10–15, and 16–18 years. although the prevalence of fungi was similar in both genders (♀32.7% and ♂30.58%), the species spectrum and occurrence of individual fungi species differed significantly among the tested groups. the occurrence of c. albicans in girls decreased compared to that of c. tropicalis and c. dubliniensis. an analysis of seasonal fungi occurrence, depending on the specificity of the tested ontocenoses and children’s age, showed that each gender is susceptible to infection by a different complex of factors, with a correlation with age being observed for girls. infections of potential fungal pathogens in generally healthy children typically end with the elimination of the fungi from the children’s bodies. according to the findings of these researchers’ studies, it happens in approx. 65% cases (ejdys, 2003a). acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 5 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) in other cases, commensalism or colonization is suspected, with possible transition to carriage. a single test of a child without any disease symptoms does not provide a basis for a clear identification of the relationship between the child’s body and a potential fungal pathogen. an inconclusive interpretation of the results of a single test of children led the researcher to conduct a distance control, whose aim was to differentiate fungal infections in healthy children into temporary and permanent, and determine the factors that predispose children to temporary fungal infections (commensalism and colonization) and carriage (ejdys, 2006). tests were conducted on smears taken twice within a 5-year span during the same season of the year from the oral cavities, pharynxes, and noses of 34 children aged 7 and 12 years. fungi were found in seven children in the first test and in nine children in the second test, and the fungi incidence was 20.6% and 26.5%, respectively, with fungi found only in three girls (8.8%) in both tests. single-focus infections were the most frequent. fungi were isolated from the ontocenosis of the oral cavity and the pharynx of one child in each test. differences in the ontocenoses diversity were observed in both tests. it was concluded that the children in whom fungi were found twice could be carriers. the physiological status of cohabitation with fungi in children with a single infection cannot be determined. however, all positive results must be treated very seriously in the context of potential future mycosis caused by yeasts (ejdys, 2006). initially, the research conducted in the school environment focused on fungal infections in children (dynowska & ejdys, 2000; ejdys, 2003a), to identify individuals particularly susceptible to infections (ejdys, 2003b), and factors that favor colonization of organ ontocenoses by various yeast species (ejdys, 2008a; ejdys & dynowska, 2004). however, in addition to individuals potentially susceptible to infections, there are other sources, ways, or vectors of fungal infections in the school environment. air is an important vector of fungi transmission between organisms (dynowska & ejdys, 1999). fungal diaspores account for the majority of bioaerosols in atmospheric air. spores of yeast-like fungi, which are heavier than those of mold fungi, occur in the bottom part of the air column. this is the height at which there are two main gateways of infection in primary school children: the mouth and nose. therefore, a high density of fungi in the air may pose a serious threat to the health of children who breathe in such air. the presence of the aforementioned fungi species in the air within and around school buildings was examined in may (heating on) and november (heating off), and the fungal features that help them survive in the school environment were analyzed (ejdys, 2008b; 2009b; ejdys et al., 2009). the air in school buildings, of the right humidity and temperature, was found to contain 28 species of 15 genera of yeast-like fungi. the atmospheric air was found to contain 20 species of 14 genera. kluyveromyces lactis dominated in the indoor air, whereas yarrowia lipolytica, which is the species with the highest symbiotic capability, dominated in the air outdoors. a comparison of the habitats under study showed, from an epidemiological perspective, that schoolrooms have greater potential to cause infections than the air outdoors. the worst yeast species spectrum was observed in intensively used rooms with limited access to light and oxygen; hence, they are the greatest threat to the health of children and school personnel. nonfermenting isolates and those that did not produce pigments dominated in heated rooms, whereas fermenting fungi and those capable of producing pigments dominated when heating was off (ejdys, dynowska, et al., 2014). the ability to produce carotenoid pigments is clearly promoted in indoor yeast. it may be associated with the addition of intermediate stages of carotenoid biosynthesis or an excess of oxidative compounds. two-thirds of the isolates did not require any vitamins for their growth. five isolates of four species exhibited acid-formation properties: dekkera anomala, d. bruxellensis, y. lipolytica, and c. tropicalis (syn. c. citrica). owing to acid production, the fungi could dissolve inorganic compounds in building structures and release essential micronutrients. these findings suggested that some yeasts inhabit rooms as their main environment, whereas some live there temporarily because the human body is their main habitat (ejdys, 2008b). acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 6 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) determination of the survival factors for the fungi outside the human ontosphere was regarded as particularly important (ejdys, 2009a; ejdys & rasztęborska, 2010). isolates of c. albicans, c. dubliniensis, and c. tropicalis obtained from human organ ontocenoses were used in the research. the strains were kept on dry glasses, with only access to the aerosol in the laboratory room, and their viability and survival rate was assessed every two weeks. a 100% death rate was not noted for any of the isolates during the 22-week observation; this proves high viability and suggests the need for improved hygiene in schoolrooms. a 4-year mycological monitoring of school rooms demonstrated that yeasts account for a third of the mycobiota in the air and on walls; a total of 772 fungal isolates of 151 species and 48 genera were obtained in the test, with 70 sterile isolates. fungi of the genera aspergillus (36 species), penicillium (26), and candida (14 + two anamorphs) were the most frequent. aspergillus fumigatus was the dominant species (126 isolates). this fungus is a permanent component of mycobiota in schools/public buildings in north-east poland. its count in rooms should be monitored continuously; this applies particularly to thermophilic isolates, owing to their environmental plasticity and potential pathogenicity (ejdys et al., 2013). such a broad spectrum of fungi species has not yet been observed in studies of indoor bioaerosols in this part of europe (ejdys, 2007a, 2007b; ejdys & biedunkiewicz, 2011; ejdys et al., 2013). it seems that it is largely a consequence of the adopted research methodology (ejdys, 2011; ejdys et al., 2015). simultaneous application of different incubation temperatures (25, 37, and 40 ◦c) allowed for the broadening of the known mycobiota spectrum in schoolrooms. sterile isolates (approx. 10%) pose an as yet undiscovered potential threat to users of the building itself, related to pathogenicity, especially allergenicity and toxin production potential, since all the 33 randomly selected fungi isolates obtained from the rooms demonstrated toxin production potential (ejdys et al., 2013). the research on the school environment conducted at the department of mycology, uwm in olsztyn resulted in the publication of 34 papers and over 40 research communications. in addition to the original papers, others devoted to research methodology, describing the methods and techniques to collect and diagnose study material more effectively, were published. 4. lichenological research the beginnings of lichenological studies in warmia and mazury date back to the late nineteenth century. the material collected in the region (the former german province of east prussia) by ohlert (1870) provided data that are still valuable as they enable researchers to assess the changes that have taken place in lichen biota over 150 years (cieśliński, 2003). after nearly 100 years, research on lichens was started at the department of botany of the academy of agriculture and technology in olsztyn (currently department of botany, uwm in olsztyn). the research was conducted by hutorowicz (e.g., hutorowicz, 1957, 1963), and subsequently, zalewska and her team (e.g., kukwa, łubek, et al., 2012; szydłowska et al., 2013; szymczyk et al., 2014, 2015; zalewska, 1998, 2012; zalewska et al., 2004a, 2004b). lichenological research at the department of mycology of the uwm in olsztyn was started in 1997 (kubiak, 1999), initially at the botany department of the teachers college in olsztyn. one of the researchers involved was professor stanisław cieśliński, an eminent polish lichenologist. affiliated to a scientific center in kielce, he worked for a brief period (1993–1994) at the botany department of the teachers college in olsztyn as a researcher and teacher. during that time, he conducted extensive field research, the results of which were published in the atlas rozmieszczenia porostów (lichenes) w polsce północno-wschodniej [distribution atlas of lichens (lichenes) in north-eastern poland] (cieśliński, 2003). the introduction to the atlas states that “the good condition of the lichen biota [in the north-east of poland] and preservation of biocenotic associations with lichens allows tracing and describing processes and relationships which – due to the anthropogenic impact, its pace and range – is not possible elsewhere in poland.” the space for such lichenological research was noted by prof. dynowska, and she initiated the acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 7 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) related actions soon after taking over the position of head of the department of mycology of the teachers college in olsztyn. dariusz kubiak was appointed head of this field of research at the department. the initial period of the research was the time when a new approach to taxonomic studies was becoming increasingly popular among lichenologists. it involved a broader use of biochemical features in species identification (guzow-krzemińska & kukwa, 2013; kubiak & kukwa, 2011), and was made possible by the development of techniques and procedures of chromatographic separation of secondary metabolites of lichenized fungi, the so-called “lichen acids” (tønsberg, 1992). owing to cooperation with other research centers, particularly with the department of plant taxonomy and nature conservation of the university of gdańsk (currently: laboratory of lichenology and experimental mycology), a laboratory of planar thin-layer chromatography (tlc) was established at the department of microbiology and mycology; this proved essential in the first research projects carried out at the department. the north-east of poland, as an area of lichenological research, is a very interesting region from a scientific perspective (cieśliński, 2003); however, it is also a highly demanding region because of the abundance of lichen biota and the number of lichen species. the initial research conducted at the department focused on exploring the taxonomic diversity, ecology, and distribution of lichen biota in the urban areas of olsztyn (kubiak, 2005). the research findings, analyzed from a bioindicative perspective, provided the basis for the doctoral thesis written by dariusz kubiak, entitled porosty olsztyna na tle antropogenicznych przekształceń środowiska [lichens in olsztyn against the background of anthropogenic transformations], with prof. dynowska as the scientific supervisor. the research, conducted for many years, documented the occurrence of over 300 lichen species within the city borders, which is a phenomenon on a global scale. (kubiak, 2005; kubiak, sucharzewska, & ejdys, 2016). the scope of the research was gradually expanded to include other areas poorly explored from a lichenological perspective, especially those regarded of key importance to lichen diversity in the region (kubiak, 2011b; kubiak, biedunkiewicz, & ejdys, 2015; kubiak, biedunkiewicz, & koźniewski, 2015; kubiak et al., 2014, 2017, 2019; kubiak & sucharzewska, 2018) in some cases, the study findings proved so interesting that they were the basis for the decision to initiate special protective measures (kubiak, biedunkiewicz, & ejdys, 2015; kubiak et al., 2014; kubiak & sucharzewska, 2013, 2018) or, in the case of protected sites, to expand the scope of protection (kubiak et al., 2017). the nature and scope of the research, as well as the specificity of the region, which stands out against other parts of the country with respect to preservation of biodiversity, resulted in the accumulation of a large amount of data on the distribution and habitat preferences of many lichen species. the data concerning a group of sterile crustose lichens, which have not been well-explored in poland, are of particular value. such data made it possible to make the first, relatively reliable assessment of the current status of the species and the danger to them (kubiak, 2010, 2011a; kubiak & łubek, 2016; kubiak & sucharzewska, 2014; kubiak & westberg, 2011; kubiak & zalewska, 2009). the discovery of new taxons of the lichen biota in poland was one of the particularly valuable results of the research (kubiak, 2003; kubiak & malicek, 2012; kubiak & westberg, 2011; kubiak & wilk, 2016; kukwa & kubiak, 2007); these include loxospora cristinae guzowkrzemińska, łubek, kubiak & kukwa (guzow-krzemińska et al., 2018). the relatively broad scope of the research was made possible by the fact that it was financed as part of the ministry of science/national centre for science research project entitled “zróżnicowanie i środowiskowe uwarunkowania lichenobioty lasów grądowych (carpinion betuli) pojezierza mazurskiego [diversity and environmental factors affecting the lichen biota in the oak-hornbeam forests (carpinion betuli) in the masurian lakeland].” at the same time, lichenological research was initiated in other parts of poland. among the more significant studies conducted at the department of mycology were those conducted in the rogów forests (kubiak & szczepkowski, 2012), mazovia (kubiak, 2009, 2013; kubiak, biedunkiewicz, & koźniewski, 2015; kubiak acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 8 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) et al., 2010), and the south of the wielkopolska region (kubiak, 2008; kubiak & biedunkiewicz, 2015). the department participated in many team projects whose aims included increasing the state of knowledge on lichen biota in selected regions of poland (fałtynowicz et al., 2015; kukwa, kowalewska, et al., 2012 ; kukwa et al., 2008), lithuania (motiejūnaitė et al., 2012), and latvia (motiejūnaitė et al., 2016). as the research at the department of microbiology and mycology intensified, it became increasingly specific. the research activities at the department focused, on the one hand, on issues related to lichen diversity, viz., methods of its assessment, principles, and ways of data interpretation (kubiak, 2012), and on the other, on the usability of the research findings in activities aimed at the preservation or sustainable exploitation of natural resources (kubiak, osyczka, & rola, 2016). many papers note the potential role and importance of old trees in the conservation of lichen diversity in forests, particularly that of stenotopic forest lichens (kubiak & osyczka, 2017; kubiak & sucharzewska, 2012, 2018). as a specific summation of this stage of lichenological research conducted at the department of microbiology and mycology, dariusz kubiak received the title of doctor habilitated of natural sciences, biological sciences discipline, based on the scientific achievement entitled “naturalne i antropogeniczne uwarunkowania różnorodności porostów epifitycznych [natural and anthropogenic factors affecting the diversity of epiphytic lichens].” the current period of lichenological research at the department of microbiology and mycology covers issues related to the identification of environmental factors that are potentially significant to species occurrence, and an assessment of their impact on the diversity of epiphytic lichens, on the site, habitat, and region/landscape scales. this research includes various types of forest communities as well as selected forms of the cultural agricultural landscape, which includes alleys of roadside trees (kubiak & osyczka, 2019, 2020; osyczka & kubiak, 2020). to understand and study the regularities of epiphytic lichen ecology, the body of environmental data, related both to the qualitative and quantitative diversity of lichen biota, as well as specific physicochemical parameters of their environment, is subjected to multidimensional statistical analyses and ordination techniques. the aim is to use this solid basis to create scientific grounds for practical measures to conserve natural resources and preserve the service that they perform for the ecosystem, which is particularly important and urgent in the face of newly emerging environmental threats. the scientific achievements of the department of microbiology and mycology in lichenological research include 70 original papers published during the past 20 years. in addition to the scientific papers, the department has accumulated extensive reference material in the form of herbarium exhibits kept at the department of microbiology and mycology (oltc). a considerable number of sterile crustose lichens are some of the remarkable items in this collection. this material has significant scientific potential. it provided the basis for many published papers written by the department staff, and is used by other researchers in taxonomic or phylogenetic studies. 5. studies of fungi – parasites of plants in the urban environment parasitic fungi are an important link in each biocenosis, and their presence in an environment depends mainly on the accessibility of host plants (majewski, 1971). finding a suitable host, which will ensure survival in different environmental structures, is the most important element of a parasite’s life strategy. a close relationship between a parasite and its host, which has developed by coevolution, produces an inseparable biological system affected by environmental conditions (mułenko, 1998). the high diversity of plant species, many of which have been introduced by humans, as well as the dynamic nature of the urban environment, make urbicenosis an interesting basis for analyzing the species diversity of parasites, studying their life strategies and the appearance of new species, and registering cases of native species being pushed out. life strategies of parasites in natural conditions, under biocenotic homeostasis, are generally well defined and relatively stable. on the other hand, under anthropopressure, all types of acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 9 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) reactions are changed frequently by disturbing factors, affecting the number and prevalence of phytopathogens. in urbicenoses, it is often a complex of transportgenerated pollution, which can be of strategic importance for the growth and development of ectoparasites, such as powdery mildew, that play an important role in adaptation. as a response to such considerations, prof. dynowska conducted long-term studies on diverse responses and the bioindicative potential of fungi of the order erysiphales in the environment under strong anthropopressure (dynowska, 1993c, 1994, 1996a, 1996b). the reconnaissance study conducted by the researcher for several years in north-east poland showed that there are erysiphales species that respond in a variety of ways to urban, particularly transport-generated, pollution. by analyzing the degree of infestation of host plants in different pollution zones, expressed by the diseases index, she identified susceptible and resistant fungi. in the case of the former, a high concentration of exhaust fumes was accompanied by a decrease in the disease index, an increase in the mycelium growth rate, and earlier maturation of the fruit body compared to those in nonurban areas. she identified the following species as susceptible: erysiphe alphitoides (microsphaera alphitoides; braun, 1987), e. cichoracearum, e. heraclei, golovinomyces sordidus (e. sordida), and podosphaera fusca (sphaerotheca fusca, s. erigerontis-caadensis; braun, 1987). in contrast, she identified the following species as resistant: erysiphe berberidis (microsphaera berberidis; braun, 1987), e. depressa, e. palczewskii (microsphaera palczewskii; braun, 1987), and e. vanbruntiana (microsphaera vanbruntiana; braun, 1987; braun & takamatsu, 2000). since there were large gaps concerning the ecology of parasitic fungi in urbicenoses in the polish literature of the subject, the findings of prof. dynowska’s research inspired other researchers to start detailed studies of their own. their aim was to observe the responses of selected erysiphales species in the urban environment, and determine the degree and scope of changes in their growth cycle. interactions in the “parasite–host” system, developed as a result of environmental changes, can be additionally modified by the presence of hyperparasites, which is why the presence of the erysiphales hyperparasite, ampelomyces quisqualis (cicinnobolus cesatii), was also taken into account in the study (sucharzewska & dynowska, 2002). seven erysiphales species were covered by the study, considering the presence of host plants in the area under study: e. alphitoides and e. hypophylla on quercus robur, e. palczewskii on caragana arborescens, g. sordidus on plantago major, podosphaera fusca on taraxacum officinale, and sawadaea tulasnei on acer platanoides l. (sucharzewska, 2007). several research tasks were adopted in order to accomplish the objective: • comparison of the degree of infestation of host plants by selected erysiphales fungi species at sites located at various distances from the transport routes in the city of olsztyn. • analysis of the share of anamorphous and telemorphous stages in the material under study as a potential adaptation index. • analysis of the adult stage – the time when chasmotetia appear, their number, and the development of rhizines, sacs, and spores – depending on the distance from transport routes. • assessment of the impact of a. quisqualis – an erysiphales hyperparasite – and other fungi that settle the thallus erysiphales species on the growth of the fungi under study, and the degree of infestation of host plants. the study findings demonstrated a higher degree of host plant infestation by all the phytopathogens selected for the study at the sites located near the main transport routes, compared to that at the control points. this led to several conclusions. in the case of e. alphitoides and e. hypophylla, a high disease index of q. robur at sites located in the zone of accumulation of transport-generated pollution was proof of the resistance of both species to this type of pollution. this applied especially to e. alphitoides, which, in the phytopathological literature, was regarded as a species sensitive to urban and industrial pollution (sucharzewska, 2009). the disease index of c. arborescens and t. officinale, infested by e. palczewskii acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 10 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) and p. fusca, respectively, was affected indirectly, but significantly, by human activity. impairment of plant health by systematic plant care treatment (lawn and hedge trimming) and toxic car exhaust probably disrupted the basic physiological processes in “plant–host” partnerships in the ecological systems under study. in consequence, the dynamic equilibrium state was disrupted for the benefit of the parasites, expressed by a high degree of infestation of host plants, of even 100% (sucharzewska & dynowska, 2005). the high disease index for a. platanoides infested by s. tulasnei on young maples, and the very low disease index or lack of parasites on mature trees, suggest that the growth of this mildew species depends mainly on the host plant age. older trees were more resistant and healthier than young ones (sucharzewska, 2010). erysiphe palczewskii was a species that stood out as one that accomplished all its life functions in the urban environment. it was here that the highest infestation degree in the host plant compared to that in the other representatives was observed, in addition to the presence of a fungus growth stage at every site, the largest number of fruiting bodies, as well as a >90% share of mature fruiting bodies with fully developed rhizines, sacs, and spores (sucharzewska & dynowska, 2005). hyperparasites were found on all the erysiphales species under study, with the largest number of them belonging to the genus ampelomyces. a strong presence of other genera was observed on the mycelium of e. alphitoides and e. palczewskii at sites near the main transport routes, e.g., alternaria, aureobasidium, cladosporium, stemphylium, and tripospermum. they reduced the number of fruiting bodies of host fungi, which proved the weaker condition of erysiphales fungi, but did not affect the degree of plant infestation (sucharzewska, dynowska, ejdys, et al., 2012). continuous research on hyperparasitism, especially an assessment of the prevalence of a. quisqualis and its impact on host growth in the urban environment, showed that this hyperparasite finds very favorable conditions for growth on the mycelium of erysiphales fungi. data obtained from the available literature showed a low share of the species in the natural environment, such as natural reserves or national parks, or even no records at all (majewski, 1971). based on the documented observations, a. quisqualis was classified as an urbanophil. the high prevalence of mycoparasites of the genus ampelomyces, with a simultaneous high prevalence of erysiphales fungi, confirmed earlier conclusions that the presence of a. quisqualis in natural conditions neither considerably limits the host plant infestation by erysiphales nor disrupts their life cycle (sucharzewska et al., 2011). ampelomyces quisqualis was found most frequently in young fruit bodies of erysiphales fungi, but with erysiphe flexuosa on aesculus hippocastanum and e. vanbruntiana var. sambuci-racemosae on sambucus racemosa; conidiospores of the hyperparasite were also found inside mature fruit bodies. this was the first description of this phenomenon. international literature only provided information on a. quisqualis settling on young fruiting bodies and transforming them into their own reproductive structures. the presence of conidiospores inside fruit bodies with properly developed rhizines suggested an improvement in the adaptive strategy (sucharzewska, dynowska, kubiak, et al., 2012). colonization of mature chasmothecia, including the completion of peridium formation with properly developed rhizines, creates proper conditions for hyperparasites for better wintering and propagation in the environment. this may result in an increase in the presence of a. quisqualis in the urban environment, and in consequence, lead to changes in the erysiphales population. an analysis of the relationship of erysiphales with other fungi (alternaria, cladosporium, stemphylium), noted in earlier studies, was also conducted. the use of electron microscope techniques (project no. 2017/01/x/nz8/00798) confirmed a parasitic relation between erysiphales spp. and the fungi present on their surface. hyperparasites destroyed the colonized development stages of erysiphales, and external and internal hyperparasitisms were observed. in the case of the former, numerous structures of parasitic fungi were found on the surface of the pathogens at the development and accompanying cell lysis stages. in the case of the latter, parasites were present inside the pathogens at the sexual and asexual stages, e.g., the presence of hyphae of stemphylium sarciniforme in the rhizines of fruit bodies of e. palczewskii (sucharzewska, 2019). acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 11 biedunkiewicz et al. / scientific research of the department of mycology (olsztyn) the development of parasitic fungi can also be affected by organisms that do not directly interact with them. several species compete for nutrients and habitat. this is the case with e. flexuosa, which infests the horse-chestnut tree and a butterfly called horse-chestnut leaf miner (cameraria ohridella), which feeds on this plant species. both species colonize the plant leaves and compete for the most beneficial living conditions. the observation of the development cycle of e. flexuosa on horsechestnut tree leaves in an urban environment, taking into account the presence of c. ohridella, led to several conclusions. a high level of presence of the species under study on two horse-chestnut species (a. hippocastanum and a. × carnea), with a distinct preference of e. flexuosa for a. × carnea, and the butterfly for a. hippocastanum, was observed. erysiphe flexuosa may have completed its life cycle, but the co-occurrence with c. ohridella affected the disease index and the number of fruit bodies formed by the fungus. analyses showed that the differences in preference of hosts ensure that e. flexuosa has proper conditions for growth and formation of a large number of fruit bodies with spores, which, as a result of the sexual process, enable the parasite to adapt to changing environmental conditions, which are particularly present in urbicenoses. therefore, it was stressed that research focusing mainly on controlling the horse-chestnut leaf miner should also take into account the presence of e. flexuosa, which, because of its invasiveness, poses a serious threat to horse-chestnut trees (sucharzewska et al., 2018). long-term monitoring conducted at the department of mycology revealed the presence of numerous parasitic fungi and fungi-like organisms on urban plants. altogether, over 300 species were identified (individual groups: pucciniales, 45%; erysiphales, 22%; and others, 33%). among the identified species, there are several regarded as interesting from the perspective of poland: entomosporium mespili, erysiphe deutziae, kuehneola uredinis, monilia linhartiana, phyllosticta juglandis, and seimatosporium lichenicola. some new hosts were found, e.g., juglans nigra for ascochyta juglandis, quercus rubra for discosia artocreas, ginkgo biloba for epicoccum purpurascens, juniperus communis for pestalotia guepinii, and crataegus monogyna for seimatosporium lichenicola. erysiphe trifolii was driven out from c. arborescens by e. palczewskii (sucharzewska et al., 2011). despite reports on the species diversity and changes of parasitic mycobiota on plants in polish cities, these issues are still poorly explored. owing to its dynamic character, urbicenosis is a perfect research ground on which numerous interactions and changes in the “parasite–host” and “host–parasite–hyperparasite” systems result in the perfection of the life strategies of parasites, with consequent preservation of the species. although studies of fungi ecophysiology in the urban environment were initiated just after the research teams at the department were formed, their dynamic growth started with the doctoral thesis entitled strategie życiowe wybranych grzybów z rzędu erysiphales w warunkach zróżnicowanej antropopresji [life strategies of selected erysiphales fungi under variable anthropopressure] (sucharzewska, 2007). the findings to date have been published in 35 papers, most of which are original scientific articles. recently, field research aimed at analyzing parasitic fungi of the phyllosphere of aquatic plants in the littoral of selected lakes in olsztyn was completed. it took into account the functioning of the fungi with respect to urban pollution and biomonitoring. references american society for testing and materials. 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(2007b). spektrum taksonomiczne i preferencje siedliskowe grzybów z rodzaju aspergillus w środowisku szkolnym [the spectrum of taxonomic and habitat preferences of aspergillus in a school environment]. in j. k. garbacz (ed.), diagnozowanie stanu środowiska: metody badawcze – prognozy: kompleksowe badania i ochrona środowiska naturalnego [diagnosing the state of the environment: research methods – forecasts: comprehensive research and protection of the natural environment] (pp. 147–158). bydgoskie towarzystwo naukowe. ejdys, e. (2008a). factors predisposing appearance of yeasts-like fungi in healthy school age girls and boys. mikologia lekarska, 15(2), 75–79. ejdys, e. (2008b). initial mycological assessment of school facilities. mikologia lekarska, 15(4), 217–222. ejdys, e. 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(1992). the sorediate and isidiate, corticolous, crustose lichens in norway. sommerfeltia, 14, 1–331. zalewska, a. (1998). remarks about some lichen species in borecka primeval forest (northeast poland). botanica lithuanica, 4(2), 157–168. zalewska, a. (2012). ecology of lichens of the puszcza borecka forest (ne poland). w. szafer institute of botany, polish academy of sciences. zalewska, a., fałtynowicz, w., krzysztofiak, a., krzysztofiak, l., & picińska-fałtynowicz, j. (2004a). porosty puszczy rominckiej [lichens of the romincka forest]. stowarzyszenie człowiek i przyroda. zalewska, a., fałtynowicz, w., krzysztofiak, a., krzysztofiak, l., & picińska-fałtynowicz, j. (2004b). porosty suwalskiego parku krajobrazowego [lichens of the suwałki landscape park]. stowarzyszenie człowiek i przyroda. acta mycologica / 2020 / volume 55 / issue 2 / article 5525 publisher: polish botanical society 19 introduction potentially pathogenic fungi isolated from waters and their application in bioindication circulation of potentially pathogenic fungi in a school environment lichenological research studies of fungi – parasites of plants in the urban environment references epicoccum nigrum link as a potential biocontrol agent against selected dermatophytes acta mycologica article id: 5516 doi: 10.5586/am.5516 publication history received: 2019-11-20 accepted: 2020-03-14 published: 2020-06-29 handling editor wojciech pusz; wrocław university of environmental and life sciences, poland; https://orcid.org/0000-00031531-2739 authors, contributions ap and ro were responsible for designing the concept, experimental structure, for data analysis, and for writing of the manuscript; kp performed the experiments, analyzed the data, and edited the manuscript; mc and md reviewed and edited the manuscript funding this research was funded by the ministry of science and higher education, university of wrocław grant for young researchers no. 0420/2557/18. competing interests no competing interests have been declared. copyright notice © the author(s) 2020. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. original research paper in plant pathology epicoccum nigrum link as a potential biocontrol agent against selected dermatophytes agata piecuch *, rafał ogórek , mariusz dyląg , magdalena cal , katarzyna przywara department of mycology and genetics, institute of genetics and microbiology, university of wrocław, poland *to whom correspondence should be addressed. email: agata.piecuch@uwr.edu.pl abstract epicoccum nigrum link is well known for producing biologically-active substances with activities against prokaryotic and eukaryotic cells. the major goal of this study was to assess e. nigrum as a potential in vitro agent against selected species of dermatophytes. the effects of the types of media used in this study on the interactions between the microscopic fungi were also examined. epicoccum nigrum’s bioactive metabolites exhibited a strong growth inhibitory effect against the dermatophytes, suggesting its potential as a biocontrol agent. notably, the strength of these interactions was dependent on the type of the medium. these secondary metabolites are not toxic against the higher eukaryotic organisms, which was further demonstrated by using the galleria mellonella model. keywords secondary metabolites; fungi; endophyte; toxicity 1. introduction epicoccum nigrum link (syn. e. purpurascens ehrenb. ex schlecht) is an endophytic fungal species, which is widely distributed as it is found on plant surfaces and in water, soil, and air. this species is particularly known for producing a variety of biologically-active substances. epicoccum nigrum isolated from the marine environment produces extracellular polysaccharides with free radical scavenging activity and is potentially useful in the prevention of oxidative damage in higher organisms (sun et al., 2011). somjaipeng et al. (2016) showed that e. nigrum could also produce taxol, which is a diterpenoid anticancer drug, and is induced by the elicitors, like water activity or ph. colored secondary metabolites, such as prodigiosins, which are also excreted by e. nigrum, may have a potential role as antimicrobial or antitumor compounds (perveen et al., 2017). epicoccum nigrum extract, which was isolated from the ferula sumbul leaves, was also found to contain prodiginine and was shown to exhibit strong antimicrobial activity against the microscopic fungi and bacteria (e.g., bacillus subtilis, escherichia coli, staphylococcus aureus, and candida albicans). it was also shown to exhibit anticancer activities against melanoma cell lines (perveen et al., 2017). this fungus, when isolated from the cambium of phellodendron amurense, has also been used for the extracellular synthesis of silver nanoparticles with a wide variety of biological activities (qian et al., 2013). epicorazines a and b, isolated from e. nigrum, exhibited antibacterial activity. cultured e. nigrum hyphae were also shown to excrete several dyes, including βand γ-carotene, rhodoxantin, and epicocconone, in the medium (baute et al., 1978). the variety of biologically-active secondary metabolites produced by e. nigrum makes it a potential organism for the biocontrol of phytopathogens. although there is one report describing the pathogenic interaction of e. nigrum with lotus acta mycologica / 2020 / volume 55 / issue 1 / article 5516 publisher: polish botanical society 1 https://doi.org/10.5586/am.5516 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0001-7791-8379 https://orcid.org/0000-0003-0564-7797 https://orcid.org/0000-0003-2062-6169 https://orcid.org/0000-0003-0446-0381 https://orcid.org/0000-0003-4482-4784 mailto:agata.piecuch@uwr.edu.pl piecuch et al. / epicoccum nigrum as a biocontrol agent corniculatus (colavolpe et al., 2018), in general, this fungus is considered to be a facultative saprotroph, exhibiting an important role in plant protection against pathogens (de cal et al., 2009). it has previously been shown that e. nigrum isolated from sugarcane inhibits several phytopathogens, such as cyanophora paradoxa and fusarium verticilloides, and is involved in enhancing the root growth (fávaro et al., 2012). although there are other reports describing the antimicrobial action of e. nigrum metabolites against yeast-like fungal human pathogens, the data on their effects against other classes of mycoses-causing fungi, such as dermatophytes, are rather limited. only one example of growth inhibition of trichophyton mentagrophytes has been described previously (mallea et al., 1991). dermatophytes are a cause of communicative diseases that are acquired from infected animals and humans. the clinical manifestations of the infections that are caused by dermatophytes includepedis and tinea capitis. the most common etiological agents causing dermatophytoses are fungal anamorphs, such as trichophyton sp. and paraphyton sp. (weitzman & summerbell, 1995). the genus trichophyton causes the infections among farm animals, mainly in calves and horses, but also in rabbits, sheep, rats, monkeys, cats, and dogs. human infections might occur after coming in contact with an infected animal. people with impaired immunological system are particularly vulnerable to these pathogens. in contrast, the genus paraphyton is comprised of anthropophilic, zoophilic, and geophilic species. among the latter, there are fungi that cause diseases in humans but are not yet reported as pathogenic fungal strains (weitzman & summerbell, 1995). it is thus important to search for new antagonists and/or biologically active substances against the dermatophytes. since higher eukaryotes are the potential hosts of these dermatophytes (achterman et al., 2011), it is crucial that the biological agents are not toxic against them, and thus in vitro and in vivo toxicity assays should be performed, preferably on mammals, prior to their application (jorjão et al., 2018). since the tests on mammals might raise several ethical issues, therefore researchers should develop other eukaryotic models, such as insects for the same. insect systems have now been extensively used to assess the virulence of fungal pathogens and for in vivo drug toxicity assays due to their low cost, easy culture, and lack of ethical restrictions. their innate immune system shares many similarities with that of mammals. among many different insect models, the greater wax moth, galleria mellonella has been frequently used by the scientific community (kavanagh & sheehan, 2018). the major goal of our study was to assess the potential of epicoccum nigrum as a biocontrol agent against dermatophytes by determining whether it shows in vitro biotic interactions with selected species of dermatophytes, as well as by assessing the effects of its secondary metabolites on the survival and growth of galleria mellonella larvae. 2. material and methods the in vitro antagonism between e. nigrum and dermatophytes were studied using the biotic series method described by mańka and mańka (1992) and ogórek and pląskowska (2011) on pda (potato dextrose agar; biocorp) and ypg (10 g/l yeast extract, 20 g/l peptone, 20 g/l glucose, 15 g/l agar) media plates, and described as an individual biotic effect. all the strains used in this study are deposited in the department of mycology and genetics, institute of genetics and microbiology, university of wrocław, poland. two rye isolates of e. nigrum, up_epc_31 and up_epc_49 (accession numbers km434173.1 and km434171.1, respectively), and four species of dermatophytes, including trichophyton tonsurans malmsten, isolated from abdominal skin, t. terrestre durie & d. frey, isolated from soil, t. mentagrophytes (c. p. robin) sabour, isolated from an ambulatory patient, and paraphyton cookei ajello ku687323.1, isolated from cave soil, were tested for the interspecies interactions. the fungal inoculates of ca. 4 mm diameter acta mycologica / 2020 / volume 55 / issue 1 / article 5516 publisher: polish botanical society 2 piecuch et al. / epicoccum nigrum as a biocontrol agent were taken from the ten days old cultures on pda, and then the mycelium was placed downwards and 2 cm apart in the center of the pda and ypg plates. each combination was prepared in four replicates. additionally, plates with mycelium of a single fungal species were used as a reference. after inoculation, the plates were incubated in the dark at 24 ± 0.5 ◦c. biotic effects of the fungi in the combined cultures were evaluated after 10 days of growth. while evaluating the biotic effects, the surrounding area of one colony that was captured by another fungal species was observed, and then the occurrence of inhibition zone between the two colonies, as well as the reduction in the colony size were considered. the appearance of each effect was scored, and points were summarized according to the scale described by mańka (1974), and the results are presented in table 1. the biotic effect induced by a particular fungal species was evaluated as an individual biotic effect (ibe). the positive effect indicates the suppression of pathogen growth, and the negative effect indicates the lack of growth suppression. the effect might be scored with the value of 0, which indicates neutral influence (mańka, 1974; ogórek & pląskowska, 2011). the size of the zone formed by e. nigrum’s colored metabolites and the size of the inhibition zones created by different e. nigrum isolates were measured. each measurement was performed in four replicates. table 1 the scale of scoring the biotic effects of the epicoccum nigrum colony on the dermatophyte colony. petri dish appearance points 0 both fungal colonies straightly abut to each other 0 i epicoccum nigrum colony abut to the dermatophyte colony in a slightly curved manner, surrounding less than 1/3 of the tested1 colony +1 ii e. nigrum colony abut to the dermatophyte colony in a slightly curved manner, surrounding at least 1/3 but no more than 1/2 of the tested colony +2 iii e. nigrum colony abut to the dermatophyte colony in a slightly curved manner, surrounding less than 1/2 but no more than 2/3 of the tested colony +3 iv e. nigrum colony abut to the dermatophyte colony in a slightly curved manner, surrounding at least 2/3 of the tested colony +4 v every mm of the inhibition zone between both colonies, caused by the e. nigrum colony +1 vi dermatophyte colony smaller by at least 1/3 but no more than 1/2 than the control colony grown on separate plate +1 vii dermatophyte colony smaller by at least 1/2 but no more than 2/3 than the control colony grown on separate plate +2 viii dermatophyte colony smaller by at least 2/3 than the control colony grown on separate plate +3 ix undeveloped dermatophyte colony +4 the points were given based on the appearance of both the colonies in a petri dish, as described by mańka (1974). 1 tested colony refers to the dermatophyte colony. additionally, the toxicity effect of e. nigrum filtrates was examined by using the galleria mellonella larvae model. sterile fungal filtrates derived from 14-day-old cultures were incubated at 25 ± 0.5 ◦c in sabouraud dextrose broth (peptone 10 g/l and glucose 40 g/l). thereafter, the caterpillars were treated with 40 µl of the sterile fungal filtrates. inoculations were performed directly into the hemocoel via the prolegs, by injections using the insulin syringes with 26g needles (fuchs et al., 2010). injections were preceded by the disinfection of the puncture sites with 70% ethanol. the inoculations with phosphate-buffered saline (pbs) and sabouraud dextrose broth were used as the experimental controls. after the injection, the larvae were incubated at 37 ± 0.5 ◦c, and the viability of the caterpillars was acta mycologica / 2020 / volume 55 / issue 1 / article 5516 publisher: polish botanical society 3 piecuch et al. / epicoccum nigrum as a biocontrol agent monitored every 24 hr consecutively for 7 days. caterpillars that were selected for the experiment were in the final instar larval stage (330 ± 30 mg in body weight). each filtrate was tested on a group of 60 individual caterpillars. the results were analyzed by one-way analysis of variance (anova) using the software package statistica version 12.0 (statsoft polska, kraków, poland). means of different test conditions were compared using the tukey’s hsd (honestly significant difference) at α ≤ 0.01. 3. results overall, both the e. nigrum isolates showed a positive biotic effect towards the tested dermatophytes, with an exception of e. nigrum up_epc_31 in the coculture with t. terrestre on ypg (table 2). the strongest biotic effect was observed for e. nigrum up_epc_31 against t. tonsurans on pda (p t. tonsurans, p. cookei = 0.003661). the same trend was observed in the case of ypg (pt. tonsurans, t. mentagrophytes = 0.009396). in the case of e. nigrum up_epc_49, all the interactions were positive, but were not significantly different on both the media plates. table 2 the individual biotic effect (ibe) between the strains of epicoccum nigrum and dermatophytes after 10 days of combined growth on pda and ypg media plates. the same experiment was performed in four independent replicates. dermatophyte species e. nigrum up_epc_31 e. nigrum up_epc_49 pda1 ypg pda ypg paraphyton cookei 5.00 ba2 1.50 abb 5.00 aa 2.50 aa trichophyton mentagrophytes 4.25 ba 0.75 bb 4.20 aa 1.25 ab trichophyton terrestre 4.00 ba −1.00 cb 2.75 aa 1.25 aa trichophyton tonsurans 7.00 aa 2.50 ab 3.75 aa 3.00 aa 1 pda (potato dextrose agar), ypg (yeast extractpeptone dextrose). 2 for each variant of the experiment, means followed by the same letter are not statistically different at α ≤ 0.01 according to tukey’s hsd test. small letters mark differences in the interaction between a particular e. nigrum isolate and the individual dermatophytes species; they refer to column means. capital letters mark the effect of media on these biotic effects within a given e. nigrum isolate and a given species of dermatophytes; they refer to row means. the biotic effects were significantly stronger on the pda plates in comparison to the ypg plates (table 2). the effect of a culture medium was specifically observed for e. nigrum up_epc_31, for which all the interactions varied in a highly significant manner (ppda, y pg = 0.000327 for t. tonsurans, ppda, y pg = 0.000349 for t. terrestre, ppda, y pg = 0.000850 for t. mentagrophytes, and ppda, y pg = 0.000643 for p. cookei). in the case of e. nigrum up_epc_49, statistically significant differences between media were recorded only for t. mentagrophytes (ppda, y pg = 0.004612) (table 2). the results of this study showed that the coculturing of one species with another species, as well as the culture medium, have an effect on the amount of pigments produced by e. nigrum and consequently, the appearance of inhibition zones (table 3). all the tested dermatophytes stimulated e. nigrum up_epc_31 to secrete the colored substances on the pda plates, with the strongest effect observed for its coculture with t. tonsurans (pt . mentaдrophytes, t. terrestre = 0.002184). in contrast, on the ypg plates, this isolate produced colored substances only when it was cocultured with p. cookei. surprisingly, e. nigrum up_epc_49 always synthesized the pigments during the coculture with different dermatophytes, regardless of the medium. moreover, there was no significant effect of the dermatophyte species on the synthesis of these colored substances by e. nigrum up_epc_49 on the ypg media, whereas on the pda media, this isolate was highly stimulated by t. terrestre (pt. terrestre, t. tonsurans = 0.000670) to produce the pigments. there was also a significant impact of the media on the amount of secreted pigments by e. nigrum up_epc_31 (ppda, y pg = 0.002048 for p. cookei, ppda, y pg = 0.000291 for t. mentagrophytes, ppda, y pg = 0.002488 for t. terrestre, and ppda, y pg = 0.000385 for t. tonsurans), as well as by e. nigrum up_epc_49 (ppda, y pg = 0.000291 for p. cookei, ppda, y pg = 0.000292 for t. mentagrophytes, and ppda, y pg = 0.006348 for t. tonsurans), with an exception in the coculture of e. nigrum up_epc_49 with t. terrestre. however, in the case of e. nigrum up_epc_31, the inhibition zones were only formed on pda plates in the cocultures with p. cookei (figure 1), acta mycologica / 2020 / volume 55 / issue 1 / article 5516 publisher: polish botanical society 4 piecuch et al. / epicoccum nigrum as a biocontrol agent table 3 the ability of epicoccum nigrum isolates to synthesize colored metabolites and create inhibition zones after 10 days of combined growth with the dermatophytes. a (+) indicates the formation of an inhibition zone, and a (–) indicates the lack of such zones. the indicated values are the average of the values from four independent experiments. dermatophyte species e. nigrum up_epc_31 e. nigrum up_epc_49 colored metabolite zone (mm) inhibition zone colored metabolite zone (mm) inhibition zone pda1 ypg pda ypg pda ypg pda ypg paraphyton cookei 7.00 aba2 1.00 ab + – 4.10 bb 7.50 aa + + trichophyton mentagrophytes 8.03 aa 0.00 ab – – 3.25 bb 7.00 aa + – trichophyton terrestre 4.00 ba 0.00 ab – – 9.00 aa 7.45 aa + – trichophyton tonsurans 8.18 aa 0.00 ab – – 5.11 bb 7.18 aa – – 1 pda (potato dextrose agar), ypg (yeast extract peptone dextrose). 2 for each variant of the experiment, means followed by the same letter are not statistically different at α ≤ 0.01 according to tukey’s hsd test. small letters mark the effect of a given species of dermatophytes on the synthesis of color metabolites by a given e. nigrum within a particular medium; they refer to column means. capital letters mark the effect of media on the synthesis of color metabolites by a given e. nigrum isolate in the combined growth with a given species of dermatophytes; they refer to row means. figure 1 an example of the inhibition zone created by the secondary metabolites secreted by epicoccum nigrum up_epc_31 (left side of the plate) to the pda medium after 10 days in the paired growth with paraphyton cookei (right side of the plate); ibe = 5.00. whereas e. nigrum up_epc_49 formed the inhibition zones in a coculture with all the dermatophytes on pda plates (besides t. tonsurans), and only with p. cookei on the ypg plates (table 3). the experiments performed to test the safety of g. mellonella larvae against the e. nigrum filtrates showed that the survival of larvae after 168 hr of incubation with the medium and the e. nigrum up_epc_31 filtrate decreased up to 96.7%, and in the case of e. nigrum up_epc_49, it decreased up to 95% (figure 2). since both, the medium and the secondary metabolites secreted by e. nigrum, reduced the survival rate of g. mellonella larvae to a similarly extent, this reduction in the viability is attributed to the medium, and not to the fungal filtrates. 4. discussion epicoccum nigrum link is a cosmopolitan fungus frequently isolated from plants, soil, or water, and is a well-known producer of various secondary metabolites (sun et al., 2011). the results obtained in our studies confirm the potential application of this species as a biocontrol agent with antimicrobial properties. the antibacterial activity of the secondary metabolites produced by e. nigrum is well documented (baute et al., 1978; perveen et al., 2017). moreover, antifungal properties of this acta mycologica / 2020 / volume 55 / issue 1 / article 5516 publisher: polish botanical society 5 piecuch et al. / epicoccum nigrum as a biocontrol agent figure 2 the effect of epicoccum nigrum up_epc_31 and up_epc_49 secondary metabolites on the survival of galleria mellonella larvae. pbs and sdb medium were used as the controls. species were also proved against numerous plant, animal, and human pathogens (mallea et al., 1991). in the present study, we demonstrated the antifungal potential of e. nigrum against dermatophytes, since inhibitory biotic interactions were observed between the e. nigrum isolates and the dermatophytes, including p. cookei, t. terrestre, t. tonsurans, and t. mentagrophytes. as shown in this study, the type of culture medium is an important factor in estimating fungal interspecies interactions (ogórek et al., 2016). pda is a medium preferable for e. nigrum, whereas dermatophytes exhibit better growth on ypg, and thus there were some differences between the strength of the interactions that were dependent on the medium. epicoccum nigrum is also a well-known producer of colored metabolites and some of them, such as prodiginine, exhibits antimicrobial properties (perveen et al., 2017). in addition, there is a correlation reported between the secretion of pigments and epicorazine a and b, by this species (baute et al., 1978). since the zones created by colored substances and biotic effects were stronger on the pda medium, we can speculate that medium composition plays an important role in stimulating interactions between e. nigrum and the dermatophytes. pda is a carbon-rich medium but is deficient in other nutrients (unlike ypg). such stress conditions might also stimulate the pigment production by e. nigrum (pradeep et al., 2013), and thus these colored substances might enhance the inhibitory biotic effects towards the dermatophytes (fatima et al., 2016). as previously reported in the literature, fungi of the genus epicoccum can also secrete bioactive metabolites with cytotoxic properties, e.g., some terpene metabolites and epicoccamide d (palacio-barrera et al., 2019). however, in this research study, we could show that e. nigrum species probably does not produce any cytotoxic substances, since the culture filtrates from e. nigrum strains did not reduce the viability of g. mellonella larvae at a significant level. in conclusion, this study is the first report describing about the antagonistic interactions between e. nigrum and dermatophytes (p. cookei, t. terrestre, t. mentagrophytes, and t. tonsurans), as well as the effects of its secondary metabolites on g. mellonella, which is an eukaryotic model organism. the results indicate towards the possible application of e. nigrum secondary metabolites for the treatment of skin dermatophytoses. therefore, in the near future, further studies will help to isolate and identify different secondary metabolites and determine their fungicidal properties. references achterman, r. r., smith, a. r., oliver, b. g., & white, t. c. 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(2013). influence of culture media on growth and pigment production by fusarium moniliforme kumbf1201 isolated from paddy field soil. world applied sciences journal, 22(1), 70–77. qian, y., yu, h., he, d., yang, h., wang, w., wan, x., & l, w. (2013). biosynthesis of silver nanoparticles by the endophytic fungus epicoccum nigrum and their activity against pathogenic fungi. bioprocess and biosystems engineering, 36(11), 1613–1619. https://doi.org/10.1007/s00449-013-0937-z somjaipeng, s., medina, a., & magan, n. (2016). environmental stress and elicitors enhance taxol production by endophytic strains of paraconiothyrium variabile and epicoccum nigrum. enzyme and microbial technology, 90, 69–75. sun, h. h., mao, w. j., jiao, j. y., xu, j. c., li, h. y., chen, y., qi, x. h., chen, y. l., xu, j., zhao, c. q., hou, y. j., & yang, y. p. (2011). structural characterization of extracellular polysaccharides produced by the marine fungus epicoccum nigrum jjy-40 and their antioxidant activities. marine biotechnology (ny), 13(5), 1048–1055. weitzman, i., & summerbell, r. c. (1995). the dermatophytes. clinical microbiology reviews, 8(2), 240–259. https://doi.org/10.1128/cmr.8.2.240 acta mycologica / 2020 / volume 55 / issue 1 / article 5516 publisher: polish botanical society 7 https://doi.org/10.7164/antibiotics.31.1099 https://doi.org/10.5197/j.2044-0588.2018.038.006 https://doi.org/10.1111/j.1365-2672.2008.04030.x https://doi.org/10.1371/journal.pone.0036826 https://doi.org/10.4161/viru.1.6.12985 https://doi.org/10.1080/21505594.2017.1397871 https://doi.org/10.3390/jof4030113 https://doi.org/10.1007/bf00436796 https://doi.org/10.1007/s00248-015-0686-4 https://doi.org/10.1016/j.btre.2019.e00308 https://doi.org/10.1016/j.micpath.2017.06.033 https://doi.org/10.1007/s00449-013-0937-z https://doi.org/10.1128/cmr.8.2.240 introduction material and methods results discussion references prof. dr hab. maria dynowska, full professor acta mycologica article id: 5523 doi: 10.5586/am.5523 publication history received: 2020-06-15 accepted: 2020-07-05 published: 2020-12-28 handling editor wojciech pusz; wrocław university of environmental and life sciences, poland; https://orcid.org/0000-00031531-2739 funding this work did not involve any funding. competing interests no competing interests have been declared. copyright notice © the author(s) 2020. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. memories and scientists prof. dr hab. maria dynowska, full professor anna biedunkiewicz * department of microbiology and mycology, faculty of biology and biotechnology, university of warmia and mazury in olsztyn, poland *to whom correspondence should be addressed. email: alibi@uwm.edu.pl professor maria dynowska (born january 23, 1949 in przemyśl) graduated from the faculty of biology and earth sciences at maria curie-skłodowska university in lublin. she obtained the following degrees: master of biology (botany) in 1973, doctor of agricultural sciences (phytopathology) in 1981 from the school of agriculture and technology in olsztyn, and doctor habilitated in biology (microbiology) in 1995 from the faculty of biology and earth sciences at the nicolaus copernicus university in toruń. she became a professor of biological sciences in 2003. although she retired in 2019, she continues her scientific work. professor maria dynowska settled in olsztyn in 1974 when she became an assistant, and then a senior lecturer, at the department of botany at the institute of biology and environmental protection at the teachers college. after obtaining the title of doctor habilitated, she assumed the position of head of the unit, which in 1996 was transformed into the unit of mycology of the teachers college and in 2004 (after the reorganization of university education in olsztyn) into the department of mycology of the university of warmia and mazury in olsztyn. two research teams were established at the initiative of professor dynowska: the team for fungi and lichens monitoring and the team for medicinal and applied mycology. this was a consequence of the implementation of earlier research plans developed at the department, oriented towards mycology in its broad sense and its links with other branches of science, especially phytosociology and geobotany, as well as phytopathology and medicine. however, the scientific accomplishments of the department staff – the majority of whom earned their doctoral degrees under the supervision of professor maria dynowska (dr hab. anna biedunkiewicz, dr hab. dariusz kubiak, dr elżbieta ejdys, and dr ewa sucharzewska) – were of the greatest importance. acta mycologica / 2020 / volume 55 / issue 2 / article 5523 publisher: polish botanical society 1 https://doi.org/10.5586/am.5523 https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://orcid.org/0000-0003-3498-7432 mailto:alibi@uwm.edu.pl biedunkiewicz / prof. dr hab. maria dynowska, full professor professor dynowska particularly valued interdisciplinary studies that focused on reservoirs and the bioecology of fungi – potential human pathogens – in different trophic systems, paying close attention to the variable nature of fungi and its consequences. the scientific achievements of professor dynowska include papers on: • crop phytopathogens (studies of genus typhula – unique in poland) and phytopathogens with affinity to the skin and blood vessels (fusarium solani, f. oxysporum); • common saprotrophs – commensals of the human skin and the alimentary tract, capable of causing severe surface and organ mycoses and lethal in extreme cases – in vulnerable patients (with cancer, tb, neutropenia, etc.); • natural reservoirs of potential anthropopathogens (e.g., aquatic ecosystems eutrophicated and polluted with household sewage, people with mycoses, fungi carriers, etc.); • the use of some microfungi (candida albicans, trichosporon beigelii, rhodotorula spp.) as indicators of water sanitary quality and epidemic threat; and • seeking to identify links of the mycosis epidemic chains (birds as mycoinfection vectors and sources). the research conducted by professor dynowska has widespread applicability to the diagnostics of mycoses of the respiratory and alimentary tracts, increasing the affinity of fungi to human tissues and the growing drug resistance of the majority of fungi isolated from clinical materials. in the latter case, professor dynowska observed a correlation with an increase in the enzymatic activity in patients on chemotherapy, radiotherapy, and antibacterial antibiotic therapy. the emphasis on medical mycology research conducted by professor dynowska was largely a consequence of her interest in medical science during her school education and studies. when she was offered an opportunity to cooperate with the independent public unit for tuberculosis and pulmonary diseases in olsztyn, she accepted it without hesitation. she initially started working as a laboratory diagnostician and then as a consultant and a supervisor of specialty training in mycoses diagnostics and prevention. the theoretical knowledge and practical experience gained at the hospital laboratory was introduced to the department by professor dynowska and applied successfully in scientific research (e.g., by doctoral students from other centers) and in teaching. it came as no surprise that a nursing unit was established at the department of mycology in 2002 under the management of professor dynowska, who hired medical staff and organized the research and teaching base of the nursing major – a novelty at the university (with european union requirements regarding educating nurses in poland). from this germinated the future faculty of medical sciences. professor dynowska was deputy dean for the major during the 2003–2007 term, which created an even stronger bond between the department and medicine and allowed for the expansion of scientific collaboration with physicians of different specialties (pulmonology, surgery, oncology, and gastroenterology). professor dynowska’s body of scientific achievement is rich and diverse. it has shone new light on the pathogenic potential of fungi that live on and in human bodies and emphasized the significant role of fungi in the environment along with bacteria. professor dynowska’s work includes 227 publications, with 135 original and review papers and 92 research communications. she has delivered 10 lectures at plenary sessions and 82 reports at domestic and foreign scientific conferences. she has also been the editor of five monographs and has prepared a number of mycologic expert opinions for public institutions and sacral facilities, as well as reviews for publishing houses and scientific periodicals. she has been awarded grants from the ministry of science, national science centre, and national centre for research and development. she has reviewed 20 doctoral theses, seven habilitation dissertations, and seven professorship proceedings. she also supervised 10 doctoral (there are two ongoing proceedings), 108 master’s, and 16 bachelor’s theses. acta mycologica / 2020 / volume 55 / issue 2 / article 5523 publisher: polish botanical society 2 biedunkiewicz / prof. dr hab. maria dynowska, full professor professor dynowska is a member of three scientific societies: the polish botanic society (since 2013: honorary member, 2007–2010: chairperson of the mycology section, three terms: chairperson of the revision committee); the polish mycological society (founding member); and the polish phytopathological society. she was a member of the mycology team at the committee of parasitology of the polish academy of sciences from 1999 to 2015 and she has been a member of the board of editors of the annals of parasitology since 2013. she was appointed a member of the committee of organismal biology of the polish academy of sciences in 2015. she has been cooperating with numerous scientific centers in poland and abroad. in 2018, she was appointed as an expert in microbiology for the polish accreditation committee. professor dynowska’s scientific capabilities, perseverance, and determined actions closely correspond with her teaching and organizational skills and control over matters important to academic circles. she has always regarded didactic matters and methods of education as important. she has frequently highlighted the fact that while trying to accumulate as many points as possible, too little time is devoted to teaching and the truth is forgotten, resulting in improper teaching, which may consequently cause the depreciation of science. professor dynowska’s major teaching-related achievements include (i) separating mycology from botany as a distinct, obligatory subject and introducing it in the following majors: biology (1990), biotechnology (2000), and microbiology (2013), and (ii) developing and adding 15 subjects in mycology to the syllabus in these majors, making it the richest offer in poland. now students express their greatest interest in phytopathology, medical mycology, applied mycology, hydromycology, lichenology, mycoses bioecology, hospital-acquired infections, edible and poisonous fungi, and mycosociology. professor dynowska’s didactic activities include all forms of teaching at the i, ii, and iii degrees of studies, comprising lectures, seminars, auditory modules, laboratory and field classes, as well as specialty workshops for students of biology, biotechnology, microbiology, and, earlier, nursing. professor dynowska’s organizational activity is indisputable. her above-mentioned achievements would not have been possible without her skills, professionalism, and commitment to this field, which has been recognized both at her parent university and elsewhere. this is demonstrated by numerous honors: 21 rector’s awards for scientific, didactic, and organizational achievements, the gold cross of merit (2000), the medal of the national education committee (2005), the honorary title “friend of the regional specialist children’s hospital in olsztyn” (2007) for outstanding commitment to the development of pediatric nursing, the statuette of st. james – the patron saint of olsztyn – on the sixty-fifth anniversary of university education in warmia and mazury for her services aimed at the development of the university of warmia and mazury in olsztyn (2010), award of the voivode of warmia and mazury named “woman with character [kobieta z charakterem]” (2010), and golden laurels of the university of warmia and mazury for her career achievements (2019). professor dynowska was also a laureate of the “golden ten” women of success in warmia and mazury (2007), although she does not view her achievements as success. she considers herself lucky to have met wise and kind teachers in her life, frequently mentioning a few names: eugenia schneider (her secondary school biology teacher) and professors józef motyka, bogusław sałata, adam paszewski, janina mikołajska (promoter of her doctoral thesis), alina skirgiełło, alicja kurnatowska, and eugenia dyner, who introduced professor dynowska to the issues of medical mycology. professor maria dynowska is generally perceived as a matter-of-fact and resolute person who highly values friendship and good manners. acta mycologica / 2020 / volume 55 / issue 2 / article 5523 publisher: polish botanical society 3 microscopic fungi on nymphaeaceae plants of the lake płociczno in drawa national park (nw poland) 1 of 11published by polish botanical society acta mycologica original research paper microscopic fungi on nymphaeaceae plants of the lake płociczno in drawa national park (nw poland) kinga mazurkiewicz-zapałowicz1*, aleksandra golianek1, łukasz łopusiewicz2 1 department of hydrobiology, ichthyology and biotechnology of reproduction, west pomeranian university of technology, szczecin, kazimierza królewicza 4, 71-550 szczecin, poland 2 center of bioimmobilisation and innovative packaging materials, west pomeranian university of technology, szczecin, klemensa janickiego 35, 71-270 szczecin, poland * corresponding author. email: kmazurkiewicz@zut.edu.pl abstract the aim of this study was to determine the occurrence of micromycetes associated with disease symptoms on the leaves and flowers of three plant species, nymphaea alba (na), nymphaea candida (nc), and nuphar lutea (nl), forming nympheid phytocoenoses on lake płociczno in drawa national park during the years 2009 to 2012. from all collected plant specimens, an overall number of 38 distinct taxa of fungi and chromistan fungal analogues was isolated. the largest diversity of taxa was found on nl (37 taxa), the lowest was on nc (4 taxa), and na contained 12 taxa. each year, anamorphic forms of ascomycota were dominant in the taxonomic structure. for the first time in poland, septoria nupharis (na, nl, nc) and colletotrichum nymphaeae (nl, nc) were found on their spotted leaves. for both of the mentioned pathogens, nymphaea candida is a new host plant in poland. botrytis cinerea, elongisporangium undulatum (= pythium undulatum), epicoccum nigrum, fusarium incarnatum (= f. semitectum), and gibberella avenacea (= fusarium avenaceum) were found each year in the studied phytocoenoses. the confirmation of na and nl flower infections by botrytis cinerea, which leads to gangrene, is an important aspect of the gray mold epidemiology. until now, the occurrence of smut fungi on nympheids in drawa national park was not observed. the taxonomic structure and the predomination of asexual stages of fungi, as well as the similarity coefficients, suggest that the seasonal decomposition of nympheids run naturally and contribute to maintaining the stability of the lake ecosystem. keywords fungal diversity; pathogenic fungi; nuphar; nymphaea; distribution introduction worldwide, nymphaeaceae salisb. consists of approx. 70 species from all over the world classified in six genera (barclaya wall., euryale salisb., nuphar sm., nymphaea l., ondinea hartog., and victoria lindl.). five taxa occur in the polish natural habitats: nymphaea alba l., n. candida c. presl. and their hybrids nymphaea ×borealis camus, nuphar lutea (l.) sibth. & sm., and n. pumila (timm) dc. the plants inhabit chemically rich (= fertile), shallow, still and slow-flowing waters, mainly on lowlands. phytocoenoses formed by these plants have been protected by the european union networking programme natura 2000 since 2010 as naturally valuable. moreover, in accordance with the decree of the polish minister of environment on 9 october 2014, nuphar pumila is strictly protected while nymphaea alba and n. candida have been doi: 10.5586/am.1079 publication history received: 2016-02-02 accepted: 2016-06-15 published: 2016-07-26 handling editor maria rudawska, institute of dendrology, polish academy of sciences, poland authors’ contributions kmz designed and conducted the research, examined the material; all authors contributed to the manuscript preparation funding research funded by the west pomeranian university of technology, szczecin (518-08030-3181-03/18). competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation mazurkiewicz-zapałowicz k, golianek a, łopusiewicz ł. microscopic fungi on nymphaeaceae plants of the lake płociczno in drawa national park (nw poland). acta mycol. 2016;51(1):1079. http:// dx.doi.org/10.5586/am.1079 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:kmazurkiewicz%40zut.edu.pl?subject=microscopic%20fungi%20on%20nymphaeaceae%20plants%20of%20the%20lake%20p%c5%82ociczno%20in%20drawa%20national%20park%20%28nw%20poland%29 http://dx.doi.org/10.5586/am.1079 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1079 http://dx.doi.org/10.5586/am.1079 2 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(1):1079 mazurkiewicz-zapałowicz et al. / microscopic fungi on nymphaeaceae plants granted partial protection. the natural habitats of these plants are used, as well as others, to estimate the quality of surface waters in the ecological state macrophyte index [1]. nymphaeaceae plants are an essential element in the process of creating an environment for other organisms. the plants health in ecosystems depends on the activity of pathogenic and saprotrofic mycobiota associated with the vegetation. in poland, phytopathological studies on nymphaeaceae plants in their natural ecosystems have so far been carried out only sporadically [2,3]. taking into consideration the shortage of knowledge on the topic of microorganisms’ participation in the functioning of valuable phytocoenosis within the nympheids, research has been set up to study the biodiversity of fungi and chromistan fungal analogues, which is crucial for the health of these plants in the protected area of drawa national park. material and methods field studies were carried out during four successive vegetation seasons (2009–2012) on fungi associated with plant species of the nymphaeaceae family: nymphaea alba l. (na), n. candida c. presl. (nc), nuphar lutea (l.) sibth. & sm. (nl). samples were collected twice during each vegetation season, depending on atmospheric conditions, i.e., at the turn of june and july and in the first decade of august from all five localities (1–5; fig. 1). each time up to 10 host plants (entire individuals when possible) with visible symptoms of diseases were collected (fig. 2a). plant nomenclature follows from mirek et al. [4]. characteristics of the studied area the lake płociczno is a flow reservoir with the płociczna river flowing through it (fig. 1). for more than 20 years, the river’s mouth has been blocked due to the increasing level of lake water forming a characteristic delta. in the 1980s, the delta of the płociczna river was a candidate for ranking as a monument of inanimate nature [5]. the lake płociczno is shallow (average depth 2.7 m) and characterized by a high concentration of nutrients dissolved in the water (eutrophic lake, at the edge of hypertrophy). the lake water has a high level of nitrates (184.1 g m2/year) and phosphates (17.0 g m2/year), and its transparency is only 1.4 m. due to the multiplicity of the water exchange (46 times a year), it is a polymictic lake [6]. an occurrence of phytocoenoses belonging to 17 plant associations was found in lake płociczno. plants belonging to the nupharo-nymphaeetum albae and nymphaeetum candidae associations form a wide belt at the opposite ends of the lake, grouped along the river banks and in the inflow and outflow areas, which are strongly shallowed by deposits of organic and inorganic matter [7]. fig. 1 the lake płociczno – localities (1–5). source: http://geoportal.gov.pl, edit. aleksandra golianek. http://geoportal.gov.pl 3 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(1):1079 mazurkiewicz-zapałowicz et al. / microscopic fungi on nymphaeaceae plants laboratory methods during the laboratory tests, fresh parts of plants with disease symptoms on the leaves, flowers, and petioles were used for the isolation of microorganisms. those plants were rinsed under cold tap water and then surface sterilized using 70% ethanol. further analysis was carried out in sterile conditions. fragments of 3 to 5 cm that contained areas of healthy tissue as well as areas with symptoms of diseases were cut from each plant. between 10 to 15 samples were obtained from each plant, depending on the intensity and diversity of the symptoms. these samples were placed on a piece of sterile filter paper under petri dishes (ø 10 and 15 cm) in humidity chambers and incubated for 2 to 21 days at a temperature of 20 ±2°c. in the meantime, the humidity chambers were regularly moistened with sterile water aerosol, and the plant samples were screened daily under a stereoscopic microscope for primary etiological symptoms. microscopic sections of infected plant tissues were prepared with lactic acid and dye methylene blue and observed under the zeiss axio microscope (125, 500, 825). in the absence of developed fungal structures, samples were transferred to plates with pda, cda, mea, and sabouraud media (merck). fungal cultures, including single-spore cultures, were prepared using standard methods as listed by waller et al. [8]. for the taxonomic identification of microscopic fungi and chromistan fungal analogues (directly from host plants as well as on artificially grown isolates) the keys by skiergiełło [9], borowska [10], ellis and ellis [11], rietmüller [12], kwaśna et al. [13], and sutton [14] were used. taxonomical system of kirk et al. [15] was adapted, verifying their nomenclature with index fungorum [16]. the calculations of biocenotic factors, such as the diversity of microscopic fungi (shannon–wiener’s coefficient – h') according to krebs [17], the structure based on taxonomic position, 5-degree frequency scale, and spatial structure [18,19] were used in the interpretation of the results. the jaccardsörensen similarity coefficient was applied for microscopic fungi occurring on host plants and for nl in the individual years of research [17]. results and discussion on three plant species from the nymphaeaceae family (na, nc, and nl) that formed compact phytocoenoses on the lake płociczno in drawa national park, 38 taxa of microscopic fungi and chromistan fungal analogues were found. among fungi ascomycetes were dominating group – 32 taxa (84.21%), while zygomycetes and basidiomycetes were represented by single species: mucor sp. and athelia rolfsii (= sclerotium rolfsii), respectively. disease symptoms were also caused by four taxa of chromistan fungal analogues from oomycota (10.52% of total species number; tab. 1). among ascomycete anamorphs (30 taxa) were dominant and their teleomorphs were formed by two species (gibberella avenacea and sordaria fumicola; fig. 2b). the dominant participation of anamorphic ascomycota forms repeated itself in subsequent years of study with 16 taxa (88.89%) in 2009, 13 taxa (92.85%) in 2010, 16 taxa (84.21%) in 2011, and 25 taxa (89.28%) in 2012. sustainability of these trends may indicate stability and a biocenotic balance of the analyzed biocenoses, which is also suggested by mułenko [20] and adamska [21]. among all studied host plants, symptoms of disease occurred only on nl and were associated with the presence of a total of 37 taxa in each year of the study. in comparison with other phytopathological-mycological studies conducted in western pomerania, it is an impressive number. in earlier studies on nl from lake glinno, only 21 taxa were listed [2], whereas on lake sitno in drawa national park only 10 taxa were found [3]. on na, the abundance of fungi and chromistan fungal analogues species in drawa national park showed smaller differences. in disease of this plant, a similar number of taxa, 12 and 14 taxa, were involved on lake płociczno and lake sitno, respectively [3]. these findings may not seem satisfactory in comparison with other regions of poland, such as lesser poland and subcarpathian voivodeships, where, on na, 42 taxa fungi and chromistan fungal analogues were found [22,23]. however, it should be noted that many species were found on phyllospheres of na grown in garden ponds. in these tanks, the cultivation of many varieties of water lilies is preferred, as they 4 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(1):1079 mazurkiewicz-zapałowicz et al. / microscopic fungi on nymphaeaceae plants are more impressive and decorative and also less resistant to pathogens, compared to their wild counterparts. in addition, in these artificial reservoirs, abiotic and biotic conditions are different from those in natural water ecosystems, which modifies the richness and species diversity of many organisms [24], including fungi and chromistan fungal analogues. the taxonomic diversity of mycobiota growing on nympheids in lake płociczno was primarily determined by a group of subrecedents (14 taxa), recedents (9 taxa), and dominants (9 taxa; tab. 1). in this study, only four species, chaetomium globosum, colletotrichum nymphaeae, fusarium sporotrichioides, septoria nupharis, were found on nc. these findings are very valuable because nc is a new host for them in poland. in all the years of the study, species diversity associated with the leaves of nymphaeaceae plants was characterized by the presence of botrytis cinerea, elongisporangium undulatum, epicoccum nigrum, fusarium incarnatum, and gibberella avenacea. gray mold pathogen was also isolated from the flowers of na and nl but was not found on nc (tab. 1). these results confirm the causal effect of b. cinerea on na and nl, which was first recognized earlier in other parts of western pomerania [2]. kowalik [22,23] and kowalik and krasny [25] have emphasized similar threats in southern poland. their research confirmed the widespread distribution of b. cinerea on na grown in garden ponds. this fact shows that the occurrence of gray mold on na and nl in poland is not just local. in many phytocoenoses, such as in these studies, it may appear as a dominant (tab. 1). however, the current lack of gray mold on nc is probably associated with the far fewer occurrences of this host plant in nympheid communities. the reasons for this change are nc withdrawal from its habitats [26] as well as na’s and nc’s ability to create hardly distinguishable hybrids. similar to other plant species’ hybrid genotypes, hybrids of na and nc may have a temporary resistance to certain physiological strains of pathogens. in this case, only changes of the pathogen’s virulence allow for the development of the disease and extend the range of host plants [27,28]. the results of this study indicate one more important aspect of the epidemiology of gray mold associated with the confirmed infection of flowers by b. cinerea. this particularly dangerous phase of the disease leads to generative organs’ gangrene, which limits the landscape and decorative value of plants with extremely ornamental flowers. another pathogen that regularly affects the health of nymphaeaceae plants is elongisporangium undulatum belonging to chromistan fungal analogues. this species and two others, septoria nupharis and colletotrichum nymphaeae, occur in large quantities and have the highest frequency (100%). elongisporangium undulatum causes extensive brown spots on the leaves that form numerous oospores starting from mid-july (fig. 2a,c). so far, this pathogen is isolated in poland, not only from the biebrza river [29], but also from nl growing in its natural habitats [2] and from na grown in artificial ponds [22,23]. the occurrence of e. undulatum (previously pythium undulatum) was also found in europe in necrotic plant tissues of nymphaea plants [30,31]. this fact shows that e. undulatum, like its relatives of the genus pythium, is responsible for the decay of hydrophytes parenchymal tissues [32]. pythium marsipium drechsler, p. pleroticum t. ito., and p. diclinum tokunaga are also involved in the biological decomposition. these species were found on nymphoides peltata [33], which forms floating leaves, similarly to nuphar and nymphaea plants. due to the occurrence on all examined host plants, three other species of fungi are noteworthy: chaetomium globosum, fusarium sporotrichioides, and septoria nupharis (tab. 1). confirmation of occurrence of s. nupharis (fig. 2d) is important, because it was probably isolated for the first time in poland [34], but brandenburger [30] mentions s. nupharis as potential pathogen of nuphar in europe. this pathogen also shows the largest share of more than 10% in the studied localities, which makes it the only eudominant in our studies (tab. 1). the importance of s. nupharis increases due to its mass occurrence on lake płociczno at a 100% frequency. a similar maximum frequency also showed colletotrichum nymphaeae, which was found for the first time in poland on nl and nc [34] but not on na (tab. 1, fig. 2e). the pathogen causes wide, soft, rotting brown spots on the edges and in the central part of the infected leaves. colletotrichum nymphaeae is the earliest known pathogen of nymphaea and nuphar, observed in portugal in 1899 and in england at the beginning of the twentieth century [35]. in 1997, c. nymphaea was confirmed as a factor contributing to the decomposition of nympheid’s leaves in the netherlands [36]. 5 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(1):1079 mazurkiewicz-zapałowicz et al. / microscopic fungi on nymphaeaceae plants fig. 2 a disease symptoms on nl’s leaves. b sordaria fimicola asci with ascospores (na). c oospores of elongisporangium undulatum (nl). d asexual spores of septoria nupharis (na). e asexual spores of colletotrichum nymphaeae (na). f,g macroconidia of gibberella avenacea (nl). h,i picnidium of phyllosticta hydrophila. j picnospores of phyllosticta hydrophila (nl). photographs by kinga mazurkiewicz-zapałowicz. 6 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(1):1079 mazurkiewicz-zapałowicz et al. / microscopic fungi on nymphaeaceae plants ta b. 1 o cc ur re nc e, fr eq ue nc y (f ) a nd d om in at io n (d ) o f f un gi s pe ci es o n n ym ph ae ac ea e of th e la ke p ło ci cz no (i n ye ar s 20 09 –2 01 2) . n o. fu ng i ph yl um n um be r of r ec or ds o f f un gi f ( % ) d (% ) n l n a n c 1 a ch ly a an dr og yn a (w . a rc he r) t .w . j oh ns on & r .l . s ey m . ( sy n. a pl an es a nd ro gy nu s) o 1 20 0. 64 sr 2 a cr em on ie lla a tr a (c or da ) s ac c. a 1 20 0. 64 sr 3 a cr em on iu m l in k (s yn . c ep ha lo sp or iu m ) a 3 40 1. 92 r 4 a lte rn ar ia a lte rn at a (f r.) k ei ss l. a 8 1 80 5. 77 d o 5 a lte rn ar ia te nu is si m a (k un ze ) w ilt sh ir e a 2 40 1. 28 r 6 a po da ch ly a pi ri fe ra z op f ( sy n. a po da ch yl a pi llu lif er a) o 1 20 0. 64 sr 7 a sp er gi llu s p . m ic he li ex h al le r a 1 20 0. 64 sr 8 a th el ia ro lfs ii (c ur zi ) c .c . t u & k im br . ( sy n. s cl er ot iu m ro lfs ii) b 1 20 0. 64 sr 9 bi po la ri s s ho em ak er a 3 40 1. 92 r 10 bo tr yt is c in er ea p er s. a 8 2 80 6. 41 d o 11 c ha et om iu m g lo bo su m k un ze a 8 1 1 80 8. 97 d o 12 c ha et os ph ae ri a ve rm ic ul ar io id es (s ac c. & r ou m .) w . g am s & h ol .-j ec h. (s yn . c hl or id iu m ch la m yd os po ru m ) a 1 20 0. 64 sr 13 c la do sp or iu m c la do sp or io id es (f re se n. ) g .a . d e v ri es a 7 80 4. 49 s 14 c la do sp or iu m h er ba ru m (p er s. ) l in k a 2 1 60 1. 92 r 15 c la st er os po ri um s ch w ei n. a 1 20 0. 64 sr 16 c ol le to tr ic hu m n ym ph ae ae (p as s. ) a a a 9 1 10 0 6. 41 d o 17 el on gi sp or an gi um u nd ul at um (h .e . p et er se n) u zu ha si , t oj o & k ak is h. (s yn . p yt hi um un du la tu m ) o 8 1 10 0 5. 77 d o 18 ep ic oc cu m n ig ru m l in k a 5 60 3. 21 s 19 fu sa ri um c ul m or um (w m .g . s m .) sa cc . a 1 20 0. 64 sr 20 fu sa ri um in ca rn at um (d es m .) sa cc . ( sy n. f us ar iu m se m ite ct um ) a 10 2 60 7. 69 d o 21 fu sa ri um o xy sp or um s ch ltd l. a 4 60 2. 56 s 7 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(1):1079 mazurkiewicz-zapałowicz et al. / microscopic fungi on nymphaeaceae plants ta b. 1 c on tin ue d n o. fu ng i ph yl um n um be r of r ec or ds o f f un gi f ( % ) d (% ) n l n a n c 22 fu sa ri um sp or ot ri ch io id es s he rb . a 5 1 1 80 7. 05 d o 23 g ib be re lla a ve na ce a r .j. c oo k (s yn . f us ar iu m a ve na ce um ) a 7 1 80 5. 13 d o 24 g ib be re lla p ul ic ar is (k un ze ) s ac c. (s yn . f us ar iu m sa m bu ci nu m ) a 4 60 2. 56 s 25 g lio m as tix m ur or um (c or da ) s . h ug he s a 1 20 0. 64 sr 26 g lo bi sp or an gi um u lti m um (t ro w ) u zu ha sh i, to jo & k ak is h. (s yn . p yt hi um u lti m um ) o 3 1 80 2. 56 s 27 m el an os po ra d am no sa (s ac c. ) l in da u (s yn . g on at ob ot ry s s im pl ex ) a 3 60 1. 92 r 28 m ic ro as cu s b re vi ca ul is s .p . a bb ot t ( sy n. s co pu la ri op si s b re vi ca ul is ) a 1 20 0. 64 sr 29 m uc or m uc ed o fr es en . z 2 40 1. 28 r 30 pe ri co ni a by ss oi de s p er s. a 2 40 1. 28 r 31 ph om a gl om er at a (c or da ) w ol le nw . & h oc ha pf el a 2 40 1. 28 r 32 ph yl lo st ic ta h yd ro ph ila s pe g. a 7 1 80 5. 13 d o 33 pi th om yc es b er k. & b ro om e a 1 20 0. 64 sr 34 se pt or ia n up ha ri s r an oj . a 10 1 1 10 0 10 .9 0 eu 35 so rd ar ia fi m ic ol a (r ob er ge e x d es m .) c es . & d e n ot . a 1 20 0. 64 sr 36 tr ic ho cl ad iu m a sp er um h ar z a 1 20 0. 64 sr 37 tr ic ho de rm a ko ni ng ii o ud em . a 1 20 0. 64 sr 38 u lo cl ad iu m c ha rt ar um (p re us s) e .g . s im m on s a 3 40 1. 92 r n um be r of re co rd s 13 8 14 4 to ta l n um be r of re co rd s 15 6 a – a sc om yc ot a; b – b as id io m yc ot a; o – o om yc ot a; z – z oo m yc ot a. d o – do m in an ts ; e u – eu do m in an ts ; r – re ce de nt s; s – s ub do m in an ts ; s r – su br ec ed en ts . 8 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(1):1079 mazurkiewicz-zapałowicz et al. / microscopic fungi on nymphaeaceae plants in our study also other species of fungi were isolated from the decaying tissues. among them were numerous facultative pathogens determining the phytosanitary condition of plants, such as: the group of species occurring in mass (frequency of 61–100%): alternaria alternata, chaetomium globosum, cladosporium cladosporioides, gibberella avenacea (fig. 2f,g), fusarium sporotrichioides, and phyllosticta hydrophila (fig. 2h–j) as well as 13 common species (frequencies of 31–60%; tab. 1, fig. 3). species occurring commonly and in large numbers are mainly polyphagic, among which the representatives of the genus fusarium deserve special attention. these fungi are nonspecialized, facultative pathogens commonly found on weakened or damaged plants [13]. their ecosystemic importance is associated with diverse chemical activity that accelerates the natural course of organic matter decomposition. previous studies indicate that more and more species previously associated with a tropical climate are involved in this process. it is evidenced by the common occurrence of fusarium incarnatum on the rotting leaves of na and nl. this pathogen is a thermophilic organism [13] that is found more and more often in poland, especially on greenhouse grown plants [37]. it is worth mentioning that another thermophilic species, athelia rolfsii, takes part in the decomposition of leaves of nl and its distribution and host plant number is probably extending due to global warming. such correlations have already been proven for other pathogenic species [38]. in this context, a systematic and constant monitoring of phytosanitary condition of nymphaeaceae plants for another thermophilic species, mainly smut fungi, should be carried out. so far, their presence has only been observed in tropical and subtropical climate zones. it has been confirmed by numerous data since 1912, when doassansia nymphaea was described in india as the cause of the discoloration of the petioles of nymphaea nauchali (= n. stellata) [39]. other studies on the spot disease (on floating leaves) of nymphaea nauchalii have documented the presence of doassansiopsis tomasii in ethiopia [40], uganda [41], and cameroon [39]. another thermophilic species of smut fungi, doassansiopsis nymphoides, previously only occasionally recorded on nymphoides rautaneni in kenya [42] and zimbabwe [43], has recently been discovered in zambia in an epidemic form [44]. affinity to the subtropical climate zone has also been showed in case of doassansiopsis ticonis, isolated from nymphaea blanda in costa rica [45] and entyloma nymphaeae (cunn.) setch. (= rhamphospora nymphaeae cunn.), which attacked nymphaea tetragoni grown in a garden pond in korea [46]. however, hitherto known distribution of these pathogens can shift due to the global warming trends promoting their spread in other climate zones. there is no data on entyloma nymphaeae or other smut fungi species occurring on plants of natural or artificial localities in poland. therefore, the suggestion made by kochman and majewski [47] that these species may one day be found in poland still awaits confirmation. despite the common occurrence of potential host plants for entyloma nymphaeae, this species has not yet been found in 36.9% frequent 34.2% common 28.9% in mass fig. 3 participation of frequency types of fungi on nymphaeaceae plants of the lake płociczno (in years 2009–2012). tab. 2 similarity coefficients of fungi and chromistan fungal analogues on nl of the lake płociczno in years 2009–2012. nuphar lutea years of research 2009 2010 2011 2012 ye ar s of r es ea rc h 2009 8 species 11 species 14 species 2010 50.0% 11 species 13 species 2011 59.5% 66.7% 14 species 2012 60.9% 61.9% 59.7% tab. 3 similarity coefficients of fungi and chromistan fungal analogues on host plants of the lake płociczno in years 2009–2012. years of research (2009–2012) host plant nymphaea alba nuphar lutea nymphaea candida h os t p la nt nymphaea alba 11 species 3 species nuphar lutea 44.9% 4 species nymphaea candida 37.5% 19.5% 9 of 11© the author(s) 2016 published by polish botanical society acta mycol 51(1):1079 mazurkiewicz-zapałowicz et al. / microscopic fungi on nymphaeaceae plants poland [34]. the lack of smut fungi pathogens in naturally valuable phytocoenosis of protected nymphaeaceae suggests the potential extension of the plants’ vegetation period and the delay of their decomposition. however, in natural ecosystems, the lack of one group of pathogens promotes the development of another. for instance, in the presented studies, a mass occurrence of colletotrichum nymphaeae and septoria nupharis was observed and played a significant role in the process of plant tissue destruction. in our research the jaccard-sörensen similarity coefficient of phytopathogens associated with nl has been estimated between 50% for the years 2009 and 2010 and 66.67% for the years 2010 and 2011 (tab. 2). the coefficient for taxa associated with particular pairs of nymphaeaceae plants is even lower at about 19.5% for nc and nl to 44.89% for na and nl (tab. 3). the similarity is low despite the fact that the three studied plant species are potential hosts for the same pathogens and grow in dense patches. this is probably due to the domination of one species (nl) in the plant communities and the abundance of fungi and chromistan fungal analogues associated with this exact host plant. low species similarity of associations of the same plant species enhances the diversity of their phytocoenoses. the similarity coefficient and a high frequency of particular pathogenic/saprotrophic species are essential for the stability of ecosystems [19]. in this context, it may be said that taxonomic structure, the asexual/sexual morphs and species frequency influence the correct natural course of the seasonal decomposition of nymphaeaceae plants and contribute to the ecological balance of the whole lake ecosystem. acknowledgments the authors would like to express their sincere thanks to the provincial nature conservation officer and the management of drawa national park for giving their consent for carrying out examinations as well as to all employees of the park for their assistance during the execution of field work. references 1. dyrektywa wodna 2000/60/we [internet]. 2000 [cited 2016 jul 5]. available from: http:// geoportal.pgi.gov.pl/css/powiaty/prawo/ue_ramowa_dyrektywa_wodna 2. mazurkiewicz-zapałowicz k, wróbel m, wolska m, silicki a. phytopathogens and saprotrophic fungi of nuphar lutea (l.) sibth. et sm. in nupharo-nymphaeetum albae tomasz. 1997 plant association of lake glinno. polish journal of environmental studies. 2006;15(5d):283–287. 3. mazurkiewicz-zapałowicz k, wróbel m, wolska m. species diversity of mycobiota on nympheids of the sitno lake in the drawno national park. in: bajkiewicz-grabowska e, borowiak d, editors. anthropogenic and natural transformations of lakes. vol. 2. gdańsk: department of limnology, university od gdańsk, polish limnological society; 2008. p. 123–126. 4. mirek z, piękoś-mirkowa h, zając a, zając m, editors. flowering plants and pteridophytes of poland – a checklist. kraków: w. szafer institute of botany, polish academy of sciences; 2002. 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(flora polska. rośliny zarodnikowe polski i ziem ościennych; vol 5). http://dx.doi.org/10.1007/bf00448415 http://dx.doi.org/10.1007/bf01981538 http://dx.doi.org/10.1007/s004420050335 http://dx.doi.org/10.3852/07-189r http://dx.doi.org/10.1016/s0953-7562(09)80726-0 http://dx.doi.org/10.5423/ppj.2010.26.3.293 http://dx.doi.org/10.5423/ppj.2010.26.3.293 abstract introduction material and methods characteristics of the studied area laboratory methods results and discussion acknowledgments references 2016-07-26t11:10:39+0100 piotr otręba lichens in the agricultural land of poland – diversity, threats, and protection: a literature review 1 of 14published by polish botanical society acta mycologica review lichens in the agricultural land of poland – diversity, threats, and protection: a literature review daria zarabska-bożejewicz* institute for agricultural and forest environment, polish academy of sciences, bukowska 19, 60-809 poznań, poland * email: zardaria@wp.pl abstract agricultural landscapes provide interesting habitats and substrates occupied by lichens. nevertheless, there are still gaps in knowledge about diversity of lichenized fungi in rural areas and factors that determine their occurrence, including anthropogenic impact. the review includes recognition of this topic in the regional context in poland and presents literature data about species diversity and habitat groups. human influences in terms of their significance for lichens disappearance as well as preservation of the lichen biota are analyzed. a list of threatened lichens found in rural areas as well as a proposal for protection of the lichen biota are given. keywords lichenized fungi; rural areas; habitat groups; anthropogenic impact introduction although studies of lichens in the agricultural land of poland are becoming increasingly important, diversity of lichenized fungi in rural areas and factors that determine their occurrence, including anthropogenic impact are still weakly recognized. there is a limited number of comprehensive investigations that focus on the lichen biota of rural areas. so far, this topic has been studied in detail during large-scale surveys in the choszczno lake district (nw poland [1,2]) and the sandr nowotomys ki (w poland [3]). however, lichenological inventories of large areas in northern, i.e., the kashuby lake district [4] and the polanowska upland [5], northwestern, i.e., western pomerania [6], and northeastern poland [7,8], also revealed the presence of many lichens species in agricultural lands. more interesting data concerning lichens in such landscape were also collected at particular locations, e.g., villages and their surroundings in some regions of poland, especially podlasie, roztocze, and lubelszczyz na (e.g., [9–35]). several articles are devoted to the occurrence of lichenized fungi on bark of fruit trees, especially in orchards [3,36–44]. lichenological studies in some protected areas, i.e., the popradzki landscape park (s poland [45]), the wiśnicko-lipnicki landscape park (s poland [46]), the chełmy landscape park (sw poland [47]), the pszczewski landscape park (w poland [48]), the wdzydzki landscape park (n poland [49]), and the suwalski landscape park (ne poland [50]) also revealed taxa growing in rural areas. more interesting lichen taxa were found in the agricultural land in gdańsk pomerania [49,51–55]. numerous papers contribute to understanding impacts of human activities on the lichen biota in the polish countryside (e.g., [3,6–8,24,30,31,34,46,56,57]). in this paper, the current knowledge about the lichenized fungi in the agricultural land and its importance for lichen diversity conservation in poland is presented based on the literature survey. the article also revises threats to lichens and their protection in a man-transformed rural landscape. doi: 10.5586/am.1076 publication history received: 2016-03-31 accepted: 2016-06-23 published: 2016-07-26 handling editor maria rudawska, institute of dendrology, polish academy of sciences, poland funding publishing of the manuscript was financially supported by the statutory funds of the institute for agricultural and forest environment, polish academy of sciences (iafe pas), poznań. competing interests no competing interests have been declared. copyright notice © the author(s) 2016. this is an open access article distributed under the terms of the creative commons attribution license, which permits redistribution, commercial and noncommercial, provided that the article is properly cited. citation zarabska-bożejewicz d. lichens in the agricultural land of poland – diversity, threats, and protection: a literature review. acta mycol. 2016;51(1):1076. http://dx.doi.org/10.5586/ am.1076 digital signature this pdf has been certified using digital signature with a trusted timestamp to assure its origin and integrity. a verification trust dialog appears on the pdf document when it is opened in a compatible pdf reader. certificate properties provide further details such as certification time and a signing reason in case any alterations made to the final content. if the certificate is missing or invalid it is recommended to verify the article on the journal website. mailto:zardaria%40wp.pl?subject=lichens%20in%20the%20agricultural%20land%20of%20poland%20%e2%80%93%20diversity%2c%20threats%2c%20and%20protection%3a%20a%20literature%20review http://dx.doi.org/10.5586/am.1076 http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/ http://dx.doi.org/10.5586/am.1076 http://dx.doi.org/10.5586/am.1076 2 of 14© the author(s) 2016 published by polish botanical society acta mycol 51(1):1076 zarabska-bożejewicz / lichens in the agricultural land of poland species diversity available data suggest a moderate level of species diversity of lichenized fungi in the agricultural land. in almost completely deforested areas in rural landscape of the choszczno lake district, 184 taxa were found [1,2], while 154 taxa were recorded in the agricultural landscape of the sandr nowotomyski [3]. in the polanowska upland 146 species were collected in open areas covered by meadows, pastures, bounds, fallow lands, and psamophilous grasslands, while in villages, settlements and gravel pits 116 species were observed [5]. the biota of particular villages and their surroundings is mainly represented by several dozen lichen species [23,24,26,29–34,44], rarely exceeding 100 taxa [27]. species diversity is related to, among others, ecological factors of the sampling localities, including their climatic conditions, the availability of suitable habitats and substrates, but also to the land use. adopted sampling strategies depending on research objectives also affect lichen diversity assessment. habitat groups of lichens a high species richness of epiphytic lichens in the agricultural land is underlined in many papers. this habitat group comprises heliophilous, nitrophilous, and coniophilous lichens mainly from the genera physcia, physconia, ramalina, and xanthoria (e.g., [3,6,8,24–27,33,45,57–59]). bark of roadside and free-standing trees, as well as phorophytes growing in mid-field afforestations and mid-field peat bogs, gardens, orchards, and country parks seem to create favorable habitats for many epiphytes in rural areas (e.g., [1–3,5,7,12,20,27,30–34,38–43,59–68]). the most frequently mentioned phorophytes in the literature are ash (fraxinus sp.), willow (salix sp.), poplar (populus sp.), maple (acer sp.), linden (tilia sp.), and alder (alnus sp.) [3,5,8,10,13,15,16,18,21,23,26,27,29,30–34,39,44,45,58,59,66,68–73]. fruit trees are a special group of phorophytes in agricultural lands. the lichen biota of fruit trees of the łącka valley (s poland) was already presented in the 1960s [36,37]. afterwards, these data were supplemented by the information from the bieszczady mountains (s poland [38,39,44]), poręba wielka in gorce [64,65], the wiśnickie foothills (s poland [46]), the commune of sławno (central poland [43]), the nowotomyska plain (w poland [3,42]), and the lower vistula valley (n poland [40,41]). numerous lichen species were found in orchards located in mountains and foothills [36,38,39], and they were sampled mainly on the bark of apple and pear trees [3,30,31,33,36,38–44,46,64]. more than 60% of threatened and protected species in vanished villages in the bieszczady mts were found on bark of fruit trees [44]. specific microclimate conditions of phorophytes are thought to promote lichen growth in old orchards. particularly, lichens are more abundant within and under tree crowns, where there is high air humidity and diffuse solar radiation [43]. on the other hand, diversity of trees and their age within an orchard, types and intensity of human activities in this agroecosystem and its surroundings, including levels of air pollution as well as general environmental conditions affect epiphytes. apple orchards feature favorable light conditions, difficult access to the water on the trunk and higher ph of the bark of malus sp. [36], and these allow for establishment and persistence of relatively rich and interesting lichen biota in nearly deforested countryside [42]. some species of lichens were observed to grow preferentially on bark of apple trees [3,42,43]. for example, bacidia rubella (hoffm.) a. massal. was sampled only [3,36,42] or mainly on this phorophyte [39] and it was relatively frequently observed in apple orchards [36,39,49,71]. this lichen species has also been recorded from bark of malus domestica borkh. in other studies dealing with rural areas [44,64,65,68]. łubek [74] observed more diverse lichen biota associated with freestanding apple trees that grow at the edge of forests. comparative data are found in reports which separately analyzed lichens growing on fruit tress (e.g., [9–11,13–15,17,24,30–32,34,37,46,50,58,59,64,65,68,71,72,74,75]); most of them were obtained during lichenological surveys in southeastern and southern part of poland. more interesting species growing on bark of fruit trees, besides b. rubella (hoffm.) a. massal., are, among others, bacidia adastra sparrius & aptroot, b. subincompta (nyl.) 3 of 14© the author(s) 2016 published by polish botanical society acta mycol 51(1):1076 zarabska-bożejewicz / lichens in the agricultural land of poland arnold, bryoria fuscescens (gyeln.) brodo & d. hawksw., b. implexa (hoffm.) brodo & d. hawksw., rinodina exigua (ach.) gray, usnea hirta (l.) weber ex f.h. wigg., and u. subfloridana stirt. [24,36,39,41–43]. wirth [76] included caloplaca cerina (hedw.) th. fr. into the group of typical and indicator species for i.a. orchards; its note is known from this type of agroecosystem in the nowotomyska plain [42]. this lichen species was frequently found in the association physcietum ascendentis ochsner 1928 occurring on roadside apple trees in łącka valley and its surroundings [37]. country parks constitute other important habitats that host interesting lichen biota [7,20,62–66,68]. altogether the presence of 87 species was confirmed in rural parks that were investigated in northeastern part of poland [20]. in the historical mansion park in poręba wielka (gorce) 73 species were discovered, including threatened and rare lichens, e.g., bacidina egenula (nyl.) arnold and sarcosagium campestre (fr.) poetsch & schied. [64,65]. among 26 species known from the historical mansion park in gogolewo village (s wielkopolska region) the most interesting ones were chaenotheca brachypoda (ach.) tibell and c. trichialis (ach.) hellb. [66]. the list of lichens of the manor park in opiniogóra górna (n mazovia) consisted of 59 species and the role of this park as a refugium of forest lichens in the agricultural land was proposed [68]. rural areas provide habitats that contribute to the expansion of many epixylous lichens. more than half of the species discovered in narew village and its environs (ne poland) were represented by epixylic lichens [29]. timber constructions are common elements of regional architecture in some parts of poland, e.g., podlasie, and they are favored by lichens associated with dead and rotting wood [29]. epixylous lichens are known to occupy fences, posts, bridges, and another man-made constructions [3,7–9,11,13–18,22,24,26,27,29–34,45,46,48,71,72,77–80]. roadside wooden crosses, being a part of the regional cultural heritage in polish rural areas, are atypical wooden substrates that are colonized by lichenized fungi [23,26,27,29]. they also grow on thatched roofs [1,9,81,82]; lipnicki [1,81,82] found mainly common and widespread species. some records of lichens are also known from similar anthropogenic substrates, i.e., wood shingles [17,18,79]. diederich et al. [83] indicated the following species lecania cyrtella (ach.) th. fr., micarea denigrata (fr.) hedl. and strangospora pinicola (a. massal.) körb. as lichens associated with wood, which prefer open areas in rural environments and also habitats enriched with nitrogen compounds. all of them were observed on timber constructions in the countryside [3,26,27,29]. micarea denigrata (fr.) hedl., as well as some other taxa found on wood in rural areas in poland, i.e., carbonicola myrmecina (ach.) bendiksby & timdal, hypogymnia physodes (l.) nyl., imshaugia aleurites (ach.) s.l.f. mey., lecanora conizaeoides nyl. ex cromb., l. saligna (schrad.) zahlbr., l. varia (hoffm.) ach., and parmeliopsis ambigua (wulfen) nyl. were included into the group consisting of species associated with wood in the agricultural landscape [84]. epilythes colonize isolated blocks and stones in open areas, also when they are a part of man-made constructions [1–8,11,19–21,25,31,33,34,39,45,52,57, 58,65,71,72,75,77,79,81,85–88]. distribution of some epilithic lichens growing on isolated blocks were recognized in rural areas in the western foreland of the białowieża forest [20]. many species recorded on similar substrates in open areas in the trójmiejski landscape park and kaszubski landscape park, i.e., fields, meadows, bounds, roadsides, included mostly calciphilous, nitrophilous, heliophilous, and coniophilous lichen species [88] and these information compiled data known earlier from the kashuby lake district [4]. numerous lichens, also coniophilous and calciphilous species, were found on isolated blocks in open areas, i.e., field roads and mid-field afforestations in the surroundings of the “długogóry” nature reserve on the myśliborskie lakeland (n poland [89]). data about species growing on anthropogenic substrates rich in calcium carbonate (e.g., walls of farm buildings, wells, bridges, and posts) [1–5, 7,8,11,18,21,23–26,30–34,39,45–48,59,62,66,70–72,75,77,78,81,87,90] indicate a positive effect of agricultural activities expanding potential habitats for lichenized fungi. among them, calciphilous species or lichens that tolerate calcium carbonate, e.g., calogaya decipiens (arnold) arup, frödén & søchting, c. pusilla (a. massal.) arup, frödén & søchting, lecanora albescens (hoffm.) branth & rostr., l. dispersa (pers.) röhl., xanthoria parietina (l.) th. fr., as well as nitrophilous species from the family physciaceae [e.g. phaeophyscia orbicularis (neck.) moberg, p. nigricans (flörke) 4 of 14© the author(s) 2016 published by polish botanical society acta mycol 51(1):1076 zarabska-bożejewicz / lichens in the agricultural land of poland moberg, physcia adscendens (fr.) h. olivier, p. caesia (hoffm.) hampe ex fürnr.] are frequently observed [23,26,27,29–32]. some interesting records of lichens are known from gravestones and crosses, especially old and not renovated, and cemetery fences [2,7,19,20,23,26,27,32,46,80,90,91]. old gravestones from the nineteenth century in the village of bociek (ne poland) have been colonized by 28 species [27]. the list of lichens found in the cemetery in bogusław (w poland) consisted of 41 epilythes [90], while the presence of 14 species was documented in the cemetery in ugoszcz (nadbużański landscape park, e poland [80]). other sacral objects also allow investigators to record numerous epilythes. one of them is the monastery complex of the cistercians in lubiąż (lower silesia, sw poland); 57 species were found here, and among them the presence of very rare taxa in poland, i.e., acarospora umbilicata bagl., diplotomma venustum (körb.) körb., lecania rabenhorstii (hepp) arnold, lecidea sarcogynoides körb., massjukiella nowakii (s.y. kondr. & bielczyk) s.y. kondr., fedorenko, s. stenroos, kärnefelt, elix, j.s. hur & a., physcia dimidiata (arnold) nyl., and xanthocarpia crenulatella (nyl.) frödén, arup & søchting [92]. terricolous lichens seem to play only a minor role among lichenized fungi growing in rural areas. there is a limited amount of information about their diversity or their occurrence in these places, and therefore it is difficult to find out species associated with the agricultural landscape. available data suggest that the lichen biota occurring on soil is less diverse and interesting species are rarely found. it can be partially explained by the fact that a large part of rural areas is covered by fields and other agroecosystems which do not allow many species to survive. for example, land use activities can prevent terricolous lichens from a successful establishment due to ploughing, fertilization, etc. in the agricultural landscape more terricolous lichens grow in psamophilous grasslands, developed on the slopes of deforested dunes and fallow lands. surveys in rural areas confirmed the presence of species mainly from the genera cetraria, cladonia, peltigera, and trapeliopsis; they were observed near roadsides, balks and fallow lands [3,11,14,19,27,70,78,81,85,93,94]. some records of lichens are known from pastures [3,65,73,95] and mid-field peat bogs commonly referred as agricultural wasteland [67]. however, both trampling, grazing, as well as fertilization induce negative influence on habitat conditions and growth of terricolous lichens, especially on pastures [3]. species from genus peltigera were sampled in meadows [95], nevertheless high humidity of soil here is indicated as the factor that limits the abundance of terricolous lichens [85]. among atypical substrates occupied by lichens in the agricultural land are, e.g., metal constructions of gates and fences, hydrants, eternity roof slates, papa, plate bitumine, plastic polyester, rubber [8,24,26,27,29–33]. matwiejuk and korobkiewicz [29] recorded candelariella aurella (hoffm.) zahlbr., lecanora albescens (hoffm.) branth & rostr., l. dispersa (pers.) röhl., phaeophyscia orbicularis (neck.) moberg, physcia caesia (hoffm.) hampe ex fürnr, p. dubia (hoffm.) lettau, and xanthoria parietina (l.) th. fr. on old abandoned tyres. some other taxa, i.e., athallia holocarpa (hoffm.) arup, frödén & søchting, candelariella aurella (hoffm.) zahlbr, c. vitellina (hoffm.) th. fr., lecanora conizaeoides nyl. ex cromb., massjukiella polycarpa (hoffm.) s.y. kondr., fedorenko, s. stenroos, kärnefelt, elix, j.s. hur & a. thell, physcia dubia (hoffm.) lettau, p. tenella (scop.) dc., and polycauliona candelaria (l.) frödén, arup & søchting were found on corroded metal parts of old farm machines [24]. threats and protection the intensive agricultural activity is indicated as one of the major threats to lichens in poland [96]. farmlands are sometimes described as “lichen desert”, where lichenized fungi disappear due to application of mineral fertilizers and agrochemicals [2,6,84]. destruction of habitats and substrates through, e.g., removal of old trees growing in orchards, roadsides or mid-field afforestations, timber constructions, isolated blocks, and smaller stones can also have negative effect on lichens and eliminate sensitive species [2,3,18,24,41,88,97]. special attention should be paid to lichens that colonize bark of free-standing and old phorophytes. nowadays, their well-preserved lichen biota with rare taxa persists in northern and northeastern part of poland [4,7,20,97]. 5 of 14© the author(s) 2016 published by polish botanical society acta mycol 51(1):1076 zarabska-bożejewicz / lichens in the agricultural land of poland in the other part of the country epiphytes of free-standing trees are declining due to gradual removal of phorophytes [97]. lichen diversity in orchards is related, among others, to type and intensity of human activities [41,42]. at present there is a trend toward denser and more intensive orchards and it poses growing threats for survival of the lichen biota in farmlands. bleaching trunks of fruit trees and rubbing of tree bark by farm animals can result in a significant loss of epiphytes in the affected agroecosystem [41]. studies conducted in the bieszczady mts revealed that localization of investigated orchards at less frequently visited areas and far away from main routes positively influences lichen diversity [39]. similar observations in the same mountain range have been made in vanished villages; investigated anthropogenic habitats were indicated as important refuges of rare lichen species [44]. according to zielińska [87], agricultural activities impoverish epilithic lichens communities on isolated blocks. disappearance of some species that inhabit boulders in open areas is associated with, e.g., their dust impregnation, which enables for colonization by nitrophilous and coniophilous species such as acarospora fuscata (nyl.) th. fr., candelariella vitellina (hoffm.) müll. arg., lecanora alpigena (ach.) cl. roux., physcia adscendens (fr.) h. olivier, and p. dubia (hoffm.) lettau [6,7]. epilythes are negatively affected by cleaning and renovation of gravestones as well as other cemetery elements [35]. air pollution, also generated by household fuel combustion is other important threat for the lichen biota [3,17,41,43]. impacts of human activity in the agricultural land are also analyzed based on observations of lichens growing on anthropogenic substrates [3,6–8,24,30,31,34,46,56,57]. more than 60% of species recorded in klewinowo and hermanówka villages represented synanthropic lichens [31,34] according to olech [98]. providing new habitats, e.g., by planting trees near roads, houses, in gardens, orchards, cemeteries enables for expansion of heliophilous, nitrophilous, and coniophilous species in the agricultural land [6,7]. many hemerophilous lichens are nitrophilous, e.g., from the genera physcia, xanthoria, and ramalina, thus they commonly colonize habitats and substrates enriched with nitrogen compounds. calciphilous lichens are other frequently recorded group in rural areas; they are found on the anthropogenic substrates that are rich in calcium carbonate, e.g., walls of farm buildings, wells, bridges, and posts. the presence of many valuable and interesting taxa in the polish countryside increases the conservation value of many areas. studies devoted to protected and threatened lichens in the agricultural land of podlasie were conducted by matwiejuk [35], and among them epiphytes and terricolous species were more frequent. their diversity was similar in forest and mid-field afforestations and near rivers. also kapek [44] paid special attention to threatened lichens during lichenological surveys in vanished villages in the bieszczady mts. among 97 recognized taxa of epiphytes, 37% represented species that had been included into the red-listed categories in poland [96]. information about threatened lichens in rural areas can be also found in papers by, e.g., lipnicki and tobolewski [2], zarabska [3], kolanko and matwiejuk [23], szymczyk and zalewska [24], matwiejuk [26–28], matwiejuk and korobkiewicz [29], kiercul [30,33], łubek and biskup [43], kowalewska et al. [51], kukwa [53–55], kubiak et al. [68], and popiel and szczepańska [92]. their records after publication of the current red list of lichens in poland in 2006 are presented in tab. 1. most of them have been put on the red list of extinct and vulnerable lichens of poland [96] in the endangered categories en, nt, and vu. numerous studies conducted in the agricultural land revealed also protected species (e.g., [3,24,26–31,35,41,43,44,49,80]). lichen conservation extends beyond species protection and it includes efforts to preserve ecosystems and landscapes at least in unimpaired conditions. considering the presence of some rare species that required better protection in jabłonka stara, lichenologist postulated to include the village into the pszczewski landscape park (w poland) [48]. records of interesting lichen species in the agricultural land in the central part of the sandr nowotomyski, especially in psamophilous grasslands in nowy tomyśl surroundings and the rich lichen biota on alders (alnus sp.) and willows (salix sp.), i.e., phorophytes that grow frequently in mid-field afforestations characteristic for the region, should support the establishment of the sandr nowotomyski landscape park [3]. 6 of 14© the author(s) 2016 published by polish botanical society acta mycol 51(1):1076 zarabska-bożejewicz / lichens in the agricultural land of poland tab. 1 bibliography of threatened lichens in poland noted in the agricultural landscape after 2006 r. species category of threatenes source of literature data acarospora umbilicata bagl. nt [92] acrocordia gemmata (ach.) a. massal. vu [44,68] alyxoria varia (pers.) ertz & tehler nt [3,42,44,68] anaptychia ciliaris (l.) körb. ex a. massal. en [24,27–29,31,68] arthonia vinosa leight. nt [5] arthothelium ruanum (a. massal.) körb. nt [31,44] athallia cerinella (nyl.) arup, frödén & søchting en [49] bacidia arnoldiana körb. nt [92] b. rubella (hoffm.) a. massal. vu [3,5,42,44,49,68] b. subincompta (nyl.) arnold en [44] biatora globulosa (flörke) fr. vu [5,49] bryoria fuscescens (gyeln.) brodo & d. hawksw. vu [5,27,29,33,49] b. implexa (hoffm.) brodo & d. hawksw. cr [41] calicium abietinum pers. vu [5] c. adspersum pers. en [5] c. glaucellum ach. vu [44] c. salicinum pers. vu [5] c. viride pers. vu [5] caloplaca cerina (hedw.) th. fr. vu [26,28,42] c. obscurella (j. lahm) th. fr. nt [24] catapyrenium squamulosum (ach.) breuss nt [26] cetraria ericetorum opiz nt [5,28,49] c. islandica (l.) ach. vu [5,27,28,49] c. sepincola (ehrh.) ach. en [5,26,27] cetrelia olivetorum (nyl.) w.l. culb. & c.f. culb. en [44] chaenotheca brachypoda (ach.) tibell en [5] c. furfuracea (l.) tibell nt [3,5] c. phaeocephala (turner) th. fr. en [49,68] c. stemonea (ach.) müll. arg. en [3] c. trichialis (ach.) hellb. nt [3,5,24,68] c. xyloxena nádv. vu [49] chrysothrix candelaris (l.) j.r. laundon cr [5] circinaria gibbosa (ach.) a. nordin, savić & tibell en [28] dibaeis baeomyces (l. f.) rambold & hertel nt [5,28] 7 of 14© the author(s) 2016 published by polish botanical society acta mycol 51(1):1076 zarabska-bożejewicz / lichens in the agricultural land of poland tab. 1 continued species category of threatenes source of literature data diplotomma alboatrum (hoffm.) flot. vu [92] d. venustum (körb.) körb. vu [92] endocarpon pusillum hedw. vu [26,28] evernia prunastri (l.) ach. nt [3,5,24,26–35,41,43,44,49,68] flavoparmelia caperata (l.) hale en [3,44] graphis scripta (l.) ach. nt [44] hypogymnia farinacea zopf vu h. tubulosa (schaer.) hav. nt [3,5,24,26,27,29,31,33–35,41,43, 44,49,68] hypotrachyna revoluta (flörke) hale en [44] lecania rabenhorstii (hepp) arnold dd [92] lecanora argentata (ach.) röhl. lc [44] l. intumescens (rebent.) rabenh. en [5,44] l. persimilis (th. fr.) arnold dd [3,24,43,49,55] l. subcarpinea szatala dd [44] lecidea sarcogynoides körb. re [92] lecidella scabra (taylor) hertel & leuckert nt [24,55] leimonis erratica (körb.) r.c. harris & lendemer nt [49] melanelia subargentifera (nyl.) essl. vu [3,5,24,26–29,44,68] melanohalea elegantula (zahlbr.) o. blanco, a. crespo, divakar, essl., d. hawksw. & lumbsch vu [5] myriolecis sambuci (pers.) clem. dd [3] normandina pulchella (borrer) nyl. en [44] ochrolechia arborea (kreyer) almb. vu [44,49] o. subviridis (høeg) erichsen vu [5,68] oxneria fallax (arnold) s.y. kondr. & kärnefelt vu [31,44] pachyphiale fagicola (arnold) zwackh vu [49] parmelina pastillifera (harm.) hale dd [44] p. tiliacea (hoffm.) hale vu [3,24,27,29,31,32,68] peltigera canina (l.) willd. vu [3,5,28,31] p. hymenina (ach.) delise dd [5,49] p. polydactylon (neck.) hoffm. dd [5] p. praetextata (flörke ex sommerf.) zopf vu [3,5,44] pertusaria coccodes (ach.) nyl. nt [5,49] p. flavida (dc.) j.r. laundon en [5] p. pertusa (l.) tuck. vu [3,5] 8 of 14© the author(s) 2016 published by polish botanical society acta mycol 51(1):1076 zarabska-bożejewicz / lichens in the agricultural land of poland tab. 1 continued species category of threatenes source of literature data phaeophyscia chloantha (ach.) moberg vu [44] p. endophoenicea (harm.) moberg en [44] p. hirsuta (mereschk.) essl. en [44] p. sciastra (ach.) moberg lc [49] physcia aipolia (ehrh. ex humb.) fürnr. nt [3,27,28,44,49] p. dimidiata (arnold) nyl. vu [41,92] physconia distorta (with.) j.r. laundon en [5,41,44,49] p. perisidiosa (erichsen) moberg en [3,24,27,28,41,42,49,68] piccolia ochrophora (nyl.) hafellner vu [24,43] placynthium nigrum (huds.) gray nt [26] pleurosticta acetabulum (neck.) elix & lumbsch en [3,5,24,27–29,49,68] porpidia cinereoatra (ach.) hertel & knoph lc [92] p. macrocarpa (dc.) hertel & a.j. schwab lc [5] pseudoschismatomma rufescens (pers.) ertz & tehler vu [44] psilolechia lucida (ach.) m. choisy lc [5,92] punctelia jeckeri (roum.) kalb dd [44] p. subrudecta (nyl.) krog vu [44] ramalina farinacea (l.) ach. vu [3,5,24,26–33,44,49,68] r. fastigiata (pers.) ach. en [5,24,26,27,29,44,68] r. fraxinea (l.) ach. en [3,5,24,26–33,49,68] r. pollinaria (westr.) ach. vu [5,26,27,29,30,44,92] rinodina exigua (ach.) gray vu [24,27] stereocaulon condensatum hoffm. vu [5] s. tomentosum th. fr. en [5] strangospora pinicola (a. massal.) körb. lc [3,5,24,27,42,49,68] tuckermanopsis chlorophylla (willd.) hale vu [3,5,26–28,30,34,35,41,49] umbilicaria polyphylla (l.) baumg. lc [5] usnea filipendula stirt. vu [27,29,30,49] u. hirta (l.) weber ex f.h. wigg. vu [3,5,26,27,29,30,32,34,35,49] u. subfloridana stirt. en [3,5] varicellaria hemisphaerica (flörke) i. schmitt & lumbsch vu [5] vulpicida pinastri (scop.) j.-e. mattsson & m.j. lai nt [3,5,31,41] xanthoparmelia mougeotii (schaer.) hale vu [5] 9 of 14© the author(s) 2016 published by polish botanical society acta mycol 51(1):1076 zarabska-bożejewicz / lichens in the agricultural land of poland the continuously increase in farming intensity can lead to the loss of habitats and substrates important for the persistence of lichenized fungi. some proposals that could support protection of lichens in the agricultural landscape are given below: ■ preservation of existing habitats which offer suitable growing conditions for lichens, restoration of degraded agroecosystems, and providing new substrates for expansions of potentially valuable and interesting species in rural areas, e.g., timber constructions, concrete, mortar and other anthropogenic substrates promoting the occurrence of epilithic lichens, especially calciphilous species, preservation of old trees in orchards, gardens, mid-field afforestations and roadsides [3,29–33]; ■ partial removal of shrubs growing around trees to increase light availability on trunks. under favorable conditions heliophilous and xerophilous lichens should be observed [3]; ■ prevention of further losing of old traditional orchards and alleys with fruit trees. in the rural landscape old orchards must be conserved because they provide unique habitats for survival of many rare, protected, and threatened species [3,39–41,44]. their role as local regulators of microclimatic conditions in deforested rural areas is also known [41]. establishment of young plantations in environs of old ones could allow for expansions of epiphytic lichens growing on old fruit trees [3]. the lichen biota of old orchards could be a source of vegetative, symbiotic diaspores, or sexual ascospores that could develop into adults on bark of younger phorophytes; ■ preservation of the cultural heritage which can be related, among others, with promotion of regional architecture. the significant use of wood for several purposes in the countryside could also provide potential habitats occupied by lichens. every effort or task undertaken to maintaining timber constructions in the rural vicinity will allow for establishment and conservation of numerous epixylous lichens; ■ the legal protection in the form of recognizing isolated blocks and old trees, also alleys of trees that host the rich lichen biota as natural monuments; ■ education of local communities by, e.g., popular science articles, brochures, and easily accessible educational boards that promote the current state of knowledge about the lichen biota, its occurrence in the agricultural landscape and proposals of its protection; ■ country parks and mid-field afforestations in nearly deforested rural areas enable for the presence of lichen associated with forest communities. kubiak et al. [68] indicated the importance of older trees, the limited occurrence of acclimated tree species, diversification of habitat conditions and lack of larger industrial plants in the vicinity of study area for preservation of the lichen biota in the investigated manor park in opiniogóra górna (n mazovia). they also suggested that introduction of new plantings into park tree stand should include phorophytes, e.g,. maple and ash that create favorable substrate conditions for lichens. it should be underlined that strategies of protection in order to prevent biodiversity loss in rural areas are mainly adjusted to needs of particular groups of organisms, e.g., birds. in fact, these recommendations should be confronted with proposals of activities undertaken to preserve other groups of organisms, e.g., lichens. it will enable to develop a consistent and effective approach to solve the problems of protection, conservation, and restoration of the natural heritage. regional differences in poland are related, among others, to land history and use, as well as the intensity of tab. 1 continued species category of threatenes source of literature data x. pulla (ach.) o. blanco, a. crespo, elix, d. hawksw. & lumbsch nt [26,27] xanthoria ulophyllodes räsänen vu [44,49,55] category of threatens according to cieśliński et al. [96]: cr – critically endangered; en – endangered; vu – vulnerable; nt – near threatened; lc – least concern; dd – data deficient. nomenclature was checked according to index fungorum [99]. 10 of 14© the author(s) 2016 published by polish botanical society acta mycol 51(1):1076 zarabska-bożejewicz / lichens in the agricultural land of poland agricultural practices. the role of these factors on the lichen biota could be recognized through gathering data from different parts of the country. improving the knowledge on lichens growing in agricultural landscapes should be pivotal for the establishment of conservation strategies. acknowledgments i would like to thank the anonymous reviewer who made valuable suggestions on the manuscript. references 1. lipnicki l. udział porostów w zbiorowiskach leśnych i na siedliskach silnie zmienionych przez człowieka. in: andrzejewski r, editor. taktyka adaptacyjna populacji i biocenoz poddanych antropopresji. warszawa: szkoła główna gospodarstwa wiejskiego – akademia rolnicza w warszawie; 1990. p. 94–113. 2. lipnicki l, tobolewski z. porosty pojezierza choszczeńskiego. acta mycol. 1992;27(1):7– 30. http://dx.doi.org/10.5586/am.1992.002 3. zarabska d. porosty w krajobrazie rolniczym sandru nowotomyskiego [phd thesis]. poznań: adam mickiewicz university; 2011. 4. fałtynowicz w, tobolewski z. the lichenized ascomycotina (ascomycetes lichenisati) of the kashuby lake district in northern poland. fragmenta floristica et geobotanica. 1989;34(3–4):445–521. 5. izydorek i. porosty (grzyby lichenizowane) wysoczyzny polanowskiej. słupskie prace biologiczne. 2010;7:51–78. 6. fałtynowicz w. porosty pomorza zachodniego. studium ekologiczno-geograficzne. gdańsk: uniwersytet gdański; 1991. 7. cieśliński s. atlas rozmieszczenia porostów (lichenes) w polsce północno-wschodniej. warszawa: białowieska stacja geobotaniczna uw; 2003. 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(phytocoenosis / supplementum cartographiae geobotanicae; vol 9). 99. index fungorum [internet]. 2016 [cited 2016 jun 16]. available from: http://www.indexfungorum.org/names/names.asp http://www.indexfungorum.org/names/names.asp http://www.indexfungorum.org/names/names.asp abstract introduction species diversity habitat groups of lichens threats and protection acknowledgments references 2016-07-26t10:37:08+0100 piotr otręba