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 Colle- totrichum 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 av- enaceum) 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 im- portant 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 habi- tats: 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 (518-08- 030-3181-03/18). 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 non- commercial, 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 oth- ers, 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 envi- ronment 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 short- age 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 locali- ties (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 in- creasing 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 con- centration of nutrients dis- solved in the water (eutrophic lake, at the edge of hypertro- phy). The lake water has a high level of nitrates (184.1 g m2/year) and phosphates (17.0 g m2/year), and its trans- parency 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 asso- ciations was found in Lake Płociczno. Plants belonging to the Nupharo-Nymphaee- tum 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 depos- its of organic and inorganic matter [7]. Fig. 1 The Lake Płociczno – localities (1–5). Source: http://geoportal.gov.pl, edit. Aleksan- dra 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 in- tensity 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 cham- bers 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 (di- rectly 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 Jaccard- Sörensen 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 asco- mycetes were dominating group – 32 taxa (84.21%), while zygomycetes and basidio- mycetes 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 as- sociated 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 chrom- istan 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, Septo- ria 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 simi- lar 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 af- fects 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 occur- rence 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 spe- cies 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 twenti- eth 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 Mac- roconidia 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 patho- gens determining the phytosanitary condition of plants, such as: the group of species occurring in mass (frequency of 61–100%): Alternaria alternata, Chaetomium globosum, Cladosporium clado- sporioides, Gibberella avenacea (Fig. 2f,g), Fusarium sporotrichioides, and Phyllosticta hydrophila (Fig. 2h–j) as well as 13 common spe- cies (frequencies of 31–60%; Tab. 1, Fig. 3). Species occurring com- monly 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 im- portance is associated with diverse chemical activity that accelerates the natural course of organic matter decomposition. Previous stud- ies 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 an- other 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, Do- assansiopsis nymphoides, previously only occasionally recorded on Nym- phoides 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 nym- phaeae (Cunn.) Setch. (= Rham- phospora nymphaeae Cunn.), which attacked Nymphaea tetragoni grown in a garden pond in Korea [46]. How- ever, 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 nym- phaeae or other smut fungi species occurring on plants of natural or ar- tificial localities in Poland. Therefore, the suggestion made by Kochman and Majewski [47] that these spe- cies 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 Septo- ria 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 as- sociated 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 sta- bility 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. 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Plant Pathol J. 2010;26(3):293. http://dx.doi.org/10.5423/ PPJ.2010.26.3.293 47. Kochman J, Majewski T. Podstawczaki (Basidiomycetes), głowniowe (Ustilaginales). Warszawa: Państwowe Wydawnictwo Naukowe; 1973. (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