THE OCCURRENCE OF MYXOMYCETES FROM A LOWLAND MONTANE FOREST AND AGRICULTURAL PLANTATIONS OF NEGROS OCCIDENTAL, WESTERN VISAYAS, PHILIPPINES JULIUS RAYNARD D. ALFARO, DONN LORENZ IVAN M. ALCAYDE, JOEL B. AGBULOS, NIKKI HEHERSON A. DAGAMAC*, THOMAS EDISON E. DELA CRUZ* DEPARTMENT OF BIOLOGICAL SCIENCES, COLLEGE OF SCIENCE, UNIVERSITY OF SANTO TOMAS, ESPAÑA, MANILA, PHILIPPINES Copyright 2014, Fine Focus all rights reserved MANUSCRIPT RECEIVED 22 SEPTEMBER, 2014; ACCEPTED 19 DECEMBER, 2014 Higher floral and faunal biodiversity is expected in multi-species-covered mountainous forests than in mono-typic agricultural plantations. To verify this supposition for cryptogamic species like the plasmodial slime molds, a rapid field survey was conducted for myxomycetes and substrates in forest floor litter and agricultural plantation were collected in Negros Occidental, Philippines. Morphological characterization identified a total of 28 species belonging to the genera Arcyria, Ceratiomyxa, Collaria, Comatricha, Craterium, Cribraria, Diderma, Didymium, Hemitrichia, Lamproderma, Physarum, Stemonitis, Trichia and Tubifera. The myxomycete species Arcyria cinerea was the only abundant species found both in the agricultural and forested areas. The majority of collected species were rarely occurring. In terms of species composition, more myxomycetes were recorded in the mountainous forest (27) compared to agricultural sites. Furthermore, aerial leaf litter collected in the forests had the highest number of records for fruiting bodies but in terms of species diversity, twigs yielded higher value based on Shannon index. Findings in this study verify that a habitat with more heterogenous plant communities yields higher species of myxomycete assemblages. This research is the first study to report myxomycetes from Negros Occidental. ABSTRACTCORRESPONDING AUTHORS *Nikki Heherson A. Dagamac nhadagamac@gmail.com Institut für Botanik und Landschaftökologie Ernst – Moritz – Arndt Universität Greifswald, Soldmannstrasse 15 D-17487 Greifswald, Germany *Thomas Edison E. dela Cruz tedelacruz@mnl.ust.edu.ph Department of Biological Sciences, College of Science, and Fungal Biodiversity and Systematics Group, Research Center for the Natural and Applied Sciences University of Santo Tomas, España 1015 Manila, Philippines KEYWORDS • abundance • amoeboid eukaryotes • diversity • fruiting body • sclerotia Myxomycetes, commonly known as true slime molds, are acellular, phagotrophic, eukaryotic organisms under the Kingdom Protista (9). These organisms have been known to exhibit both fungal and protozoan char- acteristics but through an amoeboid phase, they feed on other microorganisms, including bacteria and yeast (9). These microorganisms are distributed worldwide and usually occur on dead substrata such as bark, twigs, and dried leaves of plants (29). Several studies regarding the taxonomy and ecology of myxomycetes have been conducted but most of this research was carried out in temperate regions, such as North America (35, 36), South America (16), and Europe (8,10). Numerous studies have also been completed in the tropics, such as Costa Rica (30), Puerto Rico (23) and Mexico (17). Despite the number of the studies that were executed and the high potential of biodiversity INTRODUCTION 08 • FINE FOCUS, VOL. 1 in tropical systems, little is known about them particularly in the tropical Southeast Asia such as the Philippines. Thus, myxomycete profiling in the Philippines is still considered incomplete. As a result, there is limited knowledge on the ecology and tax- onomy of myxomycetes found in such tropical areas where there are abundant forested areas serving as excellent habitats for myxomycetes (25). Previous studies on Philippines myxomy- cetes in the late 1970s and early 1980s encom- passed the most comprehensive listing for the country (24). However, most of these publica- tions during that time were merely extensive annotated lists. But in recent years, myxo- mycete diversity and ecology studies in the Philippines has progressed, as several reports have accounted on myxomycete distribution and occurrences in several habitat types, e.g. in selected forest parks (7, 19), in coastal forests (12, 18), and lowland mountain forests (3,4). These papers surveyed different forest habitats in the Luzon main island, which is just a small por- tion of the large archipelagic geography of the Philippines. No previous reports had ever doc- umented myxomycetes on the scatter islands of the Visayas region, or reported myxomycete occurrence in large agricultural plantations in the country. As such, the findings from this research paper on myxomycete composition in lowland forests will serve as a baseline reference for the profiles of myxomycetes in a local scale of Negros Occidental, but will also contribute to understanding their distribution in the whole of the Philippines. Thus, the objectives of this paper are to (1) collect myxomycetes using opportunistic sampling methods in the field, and moist chamber cultures; (2) determine the col- lected species of myxomycetes; (3) measure the sampling strategy used in the survey; (4) assess the occurrence of each myxo- mycete species; (5) calculate the species diversity of the myxomycetes from the different substrates; and (6) compare the similarities of myxomycete assemblages between mountainous forests and agricul- tural plantations. MATERIALS AND METHODS STUDY SITES AND ITS COLLECTING LOCALITIES Field survey and substrate collection were carried out during May 2013 in Negros Occi- dental, Western Visayas. The province is part of the whole Negros Island, the third largest island in the Philippines. It is estimated that the province is approximately 375 kilometers long from north to south with basically volcanic vegetation, making its arable land ideal for cultivation of economically important crops, especially sugarcanes (http://www.negros-occ. gov.ph). Based on the Philippine Atmospher- ic, Geophysical and Astronomical Services Administration – Department of Science and Technology (PAG-ASA DOST) climatological data, the whole area is characterized as having two distinct seasons: dry from December to May and wet from June to November. Along the study area, two different habitat types, namely, a lowland forested area and an agricul- tural plantation were chosen. The descriptions of each habitat types and its collecting localities (Fig. 1) are further described below. A. Forested Areas (Mt. Kanlaon National Park, 10° 24.787N, 123° 07.982E). This area rises to a height of 2,465 m (7987 ft.) and is located in the province of Negros Occidental, Western Visayas. It is characterized by low serrated mountain ranges. The forest can be APPLIED/ENVIRONMENTAL • 09 described as a moist tropical disturbed rain- forest dominated with large dipterocarp trees and non-vascular plants. Within this forest area, six sites were randomly selected along an established 1000 m accessible forest trail that serves as a transect. B. Agricultural Area (Sugarcane Plantations, Saccharum officinarum). Only sugarcane plantations along the road in the northwest part of the province were selected for this study. Three collecting localities character- ized as dry and with extensive light expo- sure were selected to collect substrates that were subjected for moist chamber cultures. Dried substrata are ideal for the prepara- tion of moist chambers, as these substrates retain spores better. The collecting local- ities are: Silay City (SC1, 10°46’46.18”N 123°0’24.80”E), Bacolod City (SC2, 10°42’29.30”N 122°58’56.72”E), and Bago City (SC3, 10°32’37.29”N 122°51’47.51”E). FIELD COLLECTION OF MYXOMYCETE SPECIMENS Fruiting bodies of myxomycetes directly observed in the field were immediately placed in clean compartmentalized plastic collecting boxes. These specimens were brought back to the laboratory and after several days of air drying, the specimens was glued in herbarium trays and placed inside matchbox-sized herbarium boxes for permanent storage. MOIST CHAMBER PREPA- RATION FROM SUBSTRATE COLLECTED IN THE TWO HABITAT TYPES Ninety samples each of ground leaf litter (GL), aerial leaf litter (AL), twigs (TW), 60 samples each of ferns (F), and 30 samples of vines (V) from mountain forests, and 90 sugarcane leaf litter (SC) from agricultur- al plantations were collected, accounting for a total of 450 substrates used for this study. The collected substrates were placed inside dry paper bags, labeled, and trans- ported back to the laboratory. Collection of samples was done following the methods described by Stephenson (36). Samples of ground floor litter were gathered at 3-5 m regular intervals. These samples con- sisted of mixtures of leaves. Twigs < 1.0 cm in diameter in size were also collect- ed. Samples of aerial litter were collected from dead twigs or leaves still attached to branches of plants and trees. The spec- imens were wrapped gently in paper before being transported to the laboratory, where they were placed in small boxes for storage to prevent insects from getting into the samples. The samples were air dried for one week to prevent the growth of molds. To avoid pseudoreplication, a single moist chamber (MC) was prepared for each sub- strate collected. The moist chambers used consisted of disposable plastic Petri dishes, 10 cm in diameter and 4 cm deep, lined with filter paper. Samples were moistened with sterile distilled water. After 24 h, excess water was removed up to the point adequate enough for the chamber to be moist, and the pH of each of the substrate was checked using a pH meter (Sartorius PB-11). Following the incubation conditions of Dagamac et al. (3), moist chambers were kept at room temperature (22-25°C) in diffuse daylight. When necessary, a small amount of water was added to each culture to maintain moist conditions. DETERMINATION OF MYXOMYCETE SPECIES The specimens obtained from the moist chamber cultures were identified using a dissecting microscope three times per week (two day intervals) for a period of up to three months, by comparing the color, size, and structure of the myxomycete plasmodia, 10 • FINE FOCUS, VOL. 1 types of fruiting bodies (e.g. sporangium, aethalium, pseudoaethalium, and plasmodi- ocarp), and spores of myxomycetes in the descriptions stated in the standard mono- graphs of myxomycetes (21, 32). Web-based electronic databases, e.g. Eumycetozoan Proj- ect (http://slimemold.uark.edu), were also utilized for verification of some morpholog- ical features. Nomenclature used for the iden- tified myxomycetes follows the names used in Nomenyx (http://nomen.eumycetozoa. com). For specimens that could not be fully identified with strong certainty due to some malformed specimens but with distinguish- ing character enough to separate as a species, the abbreviation “cf” was used in the taxon name. All specimens listed herein are depos- ited in the myxomycete herbarium of the Fungal Biodiversity and Systematics Group of the Research Center for the Natural and Applied Sciences at the University of Santo Tomas in Manila, Philippines. EVALUATION OF DATA To evaluate the sample effort of the myxo- mycete survey in this study, an individu- al-based rarefaction curve was established. Using the rarefaction formula that computes for a number of estimators for species richness of the free downloadable program EstimateS (Version 9, Colwell 2013, 100 ran- domizations), species accumulation curves were initially constructed. In accordance with Unterseher et al. (39), the Chao2 esti- mator was then chosen as the best estimator to use and was calculated using the classical settings of EstimateS. The estimated value for the sampling effort in the study area was then determined using the formula of Ndir- itu et al. (22) by dividing the actual number of species recorded by the mean number of species expected as estimated by the Chao 2 estimator. Additionally, a hyperbolic re- gression in the form of Coleman rarefaction curve according to the Michaelis-Menten formula y = ax/(b+x), where x represents the number of samples, y is the number of species recorded and the parameter a giving an estimate for the maximum number of species to be expected at this kind of sub- strate resulting in a very close curve shape (Magurran 20) was applied to the dataset. For the assessment of species occurrence of myxomycetes, species composition was initially determined for the collection site. Occurrence refers to the frequency of the presence of a particular species of myxo- mycetes in a positive MC. A moist chamber positive for having a fruiting body of a particular species was considered as one positive collection. A collection was then considered as a single unit. The number of collections reflects the abundance of myxo- mycetes in Negros Occidental and was ex- pressed as relative abundance. The relative abundance for every species of myxomy- cetes were then calculated and reported as Abundance Index (AI) by Stephenson et al. (1993). Each species were then categorized as: (1) abundant if their relative abundance (RA) is>3% of the total collections, (2) common if RA is >1.5% but <3% of the total collections, (3) occasionally occurring if RA is >0.5% but <1.5% of the total collections, and (4) rare if the myxomycetes had an RA of < 0.5% of the total collections. To further determine the myxomycetes diversity for the different substrates, species diversity was also calculated using three different diversity indices provid- ed in Magurran (20). Shannon diversity index (HS) measures species diversity with respect to both species evenness and rich- ness. This index assumes that individuals are randomly sampled from an infinitely large community and that all species are represented in the sample (14). The Gleason Index (HG) measures the species diversity in relation to species richness. Richness is defined as the number of different species APPLIED/ENVIRONMENTAL • 11 found in a biota. Pielou’s species evenness index (E), on the other hand, quantifies how equal communities are in a given sampling area. These indices are computed as follows: Equation 1: Shannon Index of Diversity (HS) = −_i(pi ln pi), where pi = the total number of individuals in the ith species. Equation 2: Gleason Index (HG) = Np − 1/ln Ni, where Np = the total number of species and Ni = the total number of individuals in the ith species. Equation 3: Pielou’s index of species even- ness (E) E = HS/Hmax where HS = Shan- non Index of Diversity and Hmax= the maximum value of HS. The similarities of myxomycete assemblag- es between the mountainous forest and ag- ricultural plantation were also compared by Sorensen’s coefficient of community index and the Percentage Similarity index. The equation for Sorensen’s coefficient is based on the presence or absences of species. Equation 4: Coefficient of Community (CC = 2c/(a+b)), where a = total number of species in the first habitat, b = total number of species in the second habitat, and c = no. of species common to both habitat. The value of CC ranges from 0 – 1 where 0 is if there are no species present in both habitat and 1 when all species are present in both habitat. On the other hand, the Per- centage Similarity (PS) index considers not only the presence or absence of species but their relative abundance. The PS value was computed as follows: Equation 5: PS = Σ min (A, B, … X) where min = the lesser of the two percentage compositions of species A, B, C, … X in the two communities. Using the combined opportunistic sampling in the field and moist chamber culture preparation, a total of 193 records of myxomycetes were noted for this survey. From these 193 records, 42 were fruiting body records in the field and 151 were recovered either as plasmodia or fruiting body records in the moist chamber cultures. In terms of the field survey, there were no field specimens that were observed in the agricultural plantations. Moreover, a higher yield of myxomycetes was noted among moist chambers in forest litter than agricultural litter. Only two bright-spored myxo- mycetes species (Arcyria cinerea and Tubifera ferruginosa) were recorded in the sugarcane litter. A total of 32 morphospecies were identified from the 193 myxomycete records. However, four of these species were only determined to the genus level (Arcyria, Comatricha, Didymium, and Stemonitis) because they were recovered from moist chambers wherein most of the fruiting bodies were already with- ered. The list of species presented here- after has a total of 28 species belonging to 14 genera. To evaluate the sampling effort used in this study, an individual based species accumulation curve was constructed using the software esti- mates and showed that the mean Chao 2 estimator reached a constant value of 59 (Fig. 2). Using the formula of Ndiritu et al. (22) to calculate the exhaustiveness of the sampling effort for the whole study, our results gave us a computed sampling effort of 54.2% for the present study. Assessing the occurrence of the 28 determinable myxomycete species, two species were reported to be abundant, RESULTS 12 • FINE FOCUS, VOL. 1 Fig. 1. Study sites: Mt. Kanlaon National Park (forested areas) and the agricultural plantation (SC) in Negros Occidental, Western Visayas, Philippines, May 2013. namely Arcyria cinerea and Didymium ni- gripes. Nine species were common, four were occasional and 13 were reported to be occur- ring as rare (Table 1). Comparing the com- position of myxomycetes from the different substrates collected from two habitat types, 27 species were found in forested areas that were characterized to have heterogenous plant litter and only two species were accounted in the sugarcane plantations. In terms of productivity of the microhabitats tested in this study by using the moist cham- bers, 121 of the 450 MCs (27%) were positive for growth of myxomycetes either as plas- modia or as fruiting body. All of the moist chambers prepared had a relatively acidic mean pH condition. Highest percent yield APPLIED/ENVIRONMENTAL • 13 Fig. 2. Individual based species accumulation curve smoothed by Cole rarefaction for the myxomycetes collection in Negros Occidental. a b c d e f g h i j k l Fig. 3. Some representative myxomycetes collected in Negros Occidental: (a) Arcyria cinerea, (b) Arcyria denudata, (c) Ceratiomyxa fruticulosa, (d) Collaria arcyrionema, (e) Craterium leucocephalum var. cylindricum, (f) Didymium squmolosum, (g) Diderma effusum, (h) Hemitricha calyculata, (i) Physarum bogoriense, (k) Stemonitis fusca, and (l) Trichia decepiens. 14 • FINE FOCUS, VOL. 1 was observed from the aerial litter (72%), and consequently had the most number of records of myxomycetes (Table 2). The other substrates, such as the twigs and vines had the next highest number of percent yield. However, these substrates had a relatively low number of determinable records due to the fact that most of the substrates were recorded as positive culture because of the appearance of plasmo- dium during the incubation period. These MCs were unsuccessful in developing into fruiting bodies (Table 2). Lowest percentage yield (18%) was observed in sugarcane litter randomly collected in the agricultural plantations of the study area. Moreover, among the six substrates collected in the two types of habitats (forest and agricultural), twigs had the highest species diversity as calculated using Gleason index (Hg Table 1. Occurrence of myxomycetes in Negros Occidental showing the number of records accounted from the rapid field survey and the use of moist chamber cultures APPLIED/ENVIRONMENTAL • 15 Table 2. Statistics of the different substrate types used in the moist chamber =2.12) and ferns had the highest species even- ness based from Pielou’s Evenness index (E =1.00). However, using the Shannon index that considers species diversity and species even- ness, twigs gave the highest value (Hs = 0.75). Comparing the assemblages of myxomycetes between the two habitat types, a CC value of 0.08 and PS value of 0.48 were computed in the study (Table 3). These results show that species similarities between the two sites were only 8.3%, which is relatively low since the only species of myxomycetes that was present in both sites was Arcyria cinerea. DISCUSSION In terms of biodiversity, the Philippines is considered to be one of the most diverse countries in the Asia Pacific basin. Howev- er, microbial diversity assessments in the country are under-investigated. This par- ticularly holds true among the less explored fungus-like protists like the myxomycetes where a generally tropical condition would seem to be favorable for their growth and development (15). In fact, in recent years, most investigations on myxomycetes in the Philippines were concentrated only among the forest vegetation and coastal habitats of the Luzon main island (3,18). Thus, findings in this research paper are the first intensive diversity report for the Visayas group of islands of the Philippine archipelago. PRODUCTIVITY OF MYXOMYCETES IN MOIST CHAMBER CULTURES The use of a moist chamber culture in assess- ing the occurrences of myxomycetes was a vital component for this study. Current studies from arid environments in China (28) and submerged plant materials in the Big Thicket National Preserve (40) employed the usage of this technique to recover species of myxomy- cetes not easily seen on the field. In our study, a total of 450 moist chambers were prepared wherein only 27% were positive for myxomy- cete growth. After almost 15 weeks of incu- bation, 11% showed positive results for fruiting bodies, while 16% were positive for plasmodial growth. This now shows that the majority of 16 • FINE FOCUS, VOL. 1 the plasmodium specimens were not able to develop to fruiting bodies. In comparison to related local studies, a similarly low yield was observed by Dagamac et al. (5), wherein 17.5% yielded plasmodia and only 5.1% yielded fruit- ing bodies from different bark samples collect- ed from the Luzon Islands. However, Kuhn et al. (13) had a percentage yield of 51%, or 214 out of 420 moist chamber cultures containing 40% positive for plasmodia and 23% positive for fruiting bodies in six highland areas in Luzon. Substrates collected in a protected ecopark by Macabago et al. (19) had a percentage yield of 51%, or 121 out of 240 moist chambers. It seems now that most studies of myxomycetes in the Philippines that used moist chambers always supported a higher level of plasmodium yield than recovering fruiting body phenologies. Perhaps the fast dessication of most of the moist chambers during the incubation time can be a factor here, since a suitable moist environ- ment is needed to allow for the plasmodium to successfully develop into fruiting bodies. It is important to note that in doing myxomycetes biodiversity and distribution studies by means of the moist chamber technique, fruiting bod- ies are more important as compared to plasmo- dium or sclerotia, since most species identifica- tion is based on the determinable morphologies of the fruiting body (11, 31). Moreover, among the six substrates collected in our study, aerial leaf litter yielded the highest level of success. The highest productivity yield from aerial leaf litter was also recorded from other studies in the tropics, including Rojas & Stephenson (27) who reported 93% in the Coco’s Island, Costa Rica, and in a more recent comparative species listing of dela Cruz et al. (6) between substrates collected in the tropics and the temperate ecoregions. This may be attributed to specimen exposure to open air where aerial litter has a higher potential in catching spores. This supposition was already demonstrated by the studies from Schnittler & Stephenson (30) where the authors noted that slight breeze can cause the myxomycetes spores to be dispersed more than one kilometer from the starting point. Perhaps aerial litter from our study has a higher probability to trap spores dispersed by wind. SPECIES COMPOSITION OF MYXOMYCETES IN NEGROS OCCIDENTAL In this study, 28 morphospecies of myxomy- cetes were collected from lowland montane forests and sugarcane plantations in the north- ern part of Negros Occidental. This number is similar in comparison to other lowland montane vegetation area studies conducted in the Luzon main islands, including Mt. Arayat National Park (3) and in Mt. Makulot (1), which reported 30 and 28 morphospecies of myx- omycetes, respectively. Albeit this number of morphospecies is not yet reflective of the overall number of myxomycetes that can be accounted in Negros Occidental as was sug- gested by the 54.2% sampling effort for this study, findings from this research paper serve as a good starting basis for future directives in understanding the distribution of myx- omycetes in a local setting. To expand the sampling effort, it is recommended to increase the distance covered during intensive surveys and to add other substrates, i.e. barks of living deciduous trees, dung of herviborous animals, and inflorescences where myxomycetes can also thrive. In terms of species composition in the whole study area, Arcyria cinerea was noted to be the only abundant species found in both the agricultural and forest habitats, with the other species occurring relatively rarely. Stephenson (35) had the highest per- centage yield of A. cinerea in moist cham- bers (85% in the upland temperate forest of Southwestern Virginia, USA). Similar results in terms of occurrence were also obtained from the studies conducted by Ro- jas et al. (26) in the northern Neotropics and APPLIED/ENVIRONMENTAL • 17 Kuhn et al. (12) in Anda island in Pangasin- an, Philippines. Our findings now support other previous results that also showed Ar- cyria cinerea to be of cosmopolitan distri- bution worldwide, since it is widely known to be tolerant to many environments. MYXOMYCETE DISTRIBU- TION IN AGRICULTURAL LIT- TER IS MORE LIMITED THAN IN FOREST LITTER Most of the related studies on myxomy- cetes in the Philippines always used litter from the forest floor. To the best of our knowledge, the findings in this paper are the first report for the Philippines attempt- ing to evaluate myxomycetes in a sugar- cane plantation where the decaying litter and vegetation is generally specific and to compare it to the myxomycete communi- ties in forest litter where decaying litter is more heterogenous. In contrast to related studies in the Paleotropics, our findings seems to contradict the observations of Tran et al. (38), which intensively evaluated distribution of myxomycete assemblages in agricultural ground litter and the forest floor. Their results showed a relatively higher productivity among agricultural litter than the forest floor litter during both the rainy and dry seasons. Perhaps the smooth surface of the sugarcane leaf is not a favorable spore trap for other myxomy- cetes species in contrast to the pubescent surfaces of the three agricultural litters used in Thailand (banana, mango and corn plantations). Nonetheless, findings from our study supports the theory that diversities of plant communities and litter hetero- geneity (37) in a study area influence the composition of myxomycete assemblages, as evident from a higher number of myx- omycete occurrences in the forest floor litters in Negros Occidental. MYXOMYCETES FROM NEGROS OCCIDENTAL AS BASELINE INFORMATION An understanding of the distribution for myxomycetes in the Philippines is still far from complete. Many ecological factors and/ or unexplored landscapes in the coun- try still need to be investigated. Despite the findings presented in this study, it is still significant to note the limitations of a descriptive study like this are associated with the sampling efforts in collecting the substrata used in the study. The most note- worthy contribution of this paper relates to the fact that it increases the knowledge about the local ecology of myxomycetes in an ecoregion of the world where investi- gations about myxomycete diversity is still considered to be in its infancy. 18 • FINE FOCUS, VOL. 1 The authors are grateful to the Department of Environment and Natural Resources (DENR, PAMB), Negros Occidental, Region VI for the gratuitous permit granted for the collection and field survey in Mt. Kanlaon Nation Park. Moreover, JRA, DLA, JBA and TEDC are indebted to the Philippine Society of Microbiology (PSM) for the undergraduate thesis subsidy grant used for this research study. NHAD would also like to thank the Deutscher Akademischer Ausland- tausch Dienst (DAAD) for the scholarship grant. Furthermore, the authors would like to thank Angelica Rea – Maminta and Sittie Aisha Macabago for all the technical assistance and to the anonymous reviewers that helped in improving this manuscript. ACKNOWLEDGEMENTS REFERENCES 1. Cheng, C.B.T., Yu, K.N.T., Campos, M.L., Adora, J.M.V., Pascua, G.C.P., Pangilinan, M.V.B., Buaya, A.T. & dela Cruz T.E.E. 2013. Occurrence and diversity of of myxomycetes (plasmodial slime molds) along the northern slope of Mt. Makulot, Cuenca, Batangas, Philippines. Asian J. Biodivers. 4:65-83. 2. Colwell, R.K. 2013. EstimateS: Statistical estimation of species richness and shared species from samples. Version 7. User’s Guide and application published at http://purl.oclc.org/estimates (accessed 23.01.2014). 3. Dagamac, N.H.A., Stephenson, S.L., & dela Cruz, T.E.E. 2014. The occurrence of litter myxomycetes at different elevations in Mt. Arayat, National Park, Pampanga, Philippines. Nova Hedwigia 98:187-196. 4. Dagamac, N.H.A., Stephenson, S.L., & dela Cruz, T.E.E. 2012. Occurrence, distribution and diversity of myxomycetes (plasmodial slime molds) along two transects in Mt. Arayat National Park, Pampanga, Philippines. Mycology 3:119-126. 5. Dagamac, N.H.A., Leontyev, D.V., & dela Cruz, T.E.E. 2010. Corticolous myxomycetes associated with Samanea samans (Jacq.) Merr. collected from different sites in Luzon Island, Philippines. The Philippine Biota 43:2-15. 6. dela Cruz, T.E.E., Rea, M.A.D., Tran, H.T.M., Ko Ko, T.W. & Stephenson, S.L. 2014. A comparative species listing of myxomycetes from tropical (Philippines) and temperate (United States) forests. Mycosphere 5:299-311. 7. dela Cruz, T.E.E., Pangilinan, M.V.B., Cruz, R.J., de Jesus, E.E., Puylong, R.G., & Dagamac, N.H.A. 2010. A checklist of plasmodial myxomycetes (slime molds) from Subic Watershed Forest Reserve, Zambales, Philippines. Acta Manilana 58:41-45. 8. Eliasson, U. 1981. Patterns of occurrence of myxomycetes in a spruce forest in South Sweden. Holarctic Ecol. 4:20-31. 9. Ing, B. 1994. The Phytosociology of Myxomycetes. New Phytol. 162:175-201. 10. Ing, B. 1983. A ravine association of Myxomycetes. J. Biogeogr. 10:299-306. 11. Ko Ko, T.W., Stephenson, S.L., Jeewon, R., Lumyong, S., & Hyde K.D. 2009. Molecular diversity of myxomycetes associated with decaying wood and forest floor leaf litter. Mycologia 63:901-906. 12. Kuhn, R.V., Javier, A.O.M., Rodillas, C.P., Parra, C.M., Corpuz, L.H.M., Buaya, A.T. & dela Cruz, T.E.E. 2013a. Diversity of plasmodial myxomycetes from Anda Island, Pangasinan, Philippines. Biotropia 20:1-9. 13. Kuhn, R.V., Javier, A.O.M., Rodillas, C.P., Parra, C.M., Corpus, L.H.M., Moron, L.S., & dela Cruz, T.E.E. 2013b. Occurrence and distribution of myxomycetes (plasmodial slime molds) in three provinces of Luzon Island, Philippines. Phil. Sci. Lett. 6:1-7. 14. Kumar, S.S.D., & Hyde, K. 2004. Biodiversity and tissue recurrence of endophytic fungi in Tripterygium wilfordii. Fungal Divers. 17:69-90. 15. Lado, C., & Wrigley de Basanta, D. 2008. A Review of Neotropical Myxomycetes (1828-2008). Anales Jard. Bot. Madrid. 65:211-254. 16. Lado, C., Estrada-Torres, A., & Stephenson, S.L. 2007. Myxomycetes collected in the first phase of a north- south transect of Chile. Fungal Divers. 25:81-101. 17. Lado, C., Estrada-Torres, A., Stephenson, S.L., Wrigley De Basanta, D., & Schnittler, M. 2003. Biodiversity assessment of myxomycetes form two tropical forest reserves in Mexico. Fungal Divers. 12:67-110. 18. Macabago, S.A.B., dela Cruz, T.E.E., & Stephenson, S.L. 2012. First records of myxomycetes from Lubang Island, Occidental Mindoro, Philippines. Sydowia 64:109–118. 19. Macabago, S.A.B., Dagamac, N.H.A., & dela Cruz, T.E.E. 2010. Diversity and distribution of plasmodial mxyomycetes (slime molds) from La Mesa Ecopark, Quezon City, Philippines. Biotropia 17:51-61. 20. Magurran, A. E. 2004. Measuring Biological Diversity. Blackwell Publishers., Oxford, UK. 21. Martin, G.W., & Alexopoulos, C.J. 1969. The Myxomycetes. University of Iowa Press, Iowa City. 22. Ndiritu, G.G., Spiegel, F.W., & Stephenson, S.L. 2009. Distribution and ecology of the assemblages of myxomycetes associated with major vegetation types in Big Bend National Park, USA. Fungal Ecol. 2:168-183. 23. Novozhilov, Y.K., Shnittler, M., Rollins, A.W., & Stephenson, S.L. 2001. Myxomycetes in different forest sites of Puerto Rico. Mycotaxon 77:285-299. 24. Reynolds, D. R. 1981.Southeast Asian myxomycetes II. Philippines. Phil. J. Bio. 10:127-150. 25. Rojas, C., & Stephenson, S.L. 2012. Rapid assessment of the distribution of myxomycetes in a southwestern Amazon forest. Fungal Ecol. 5:726-733. 26. Rojas, C., Stephenson, S.L., & Huxel, G.R. 2011. Macroecology of high elevation myxomycetes assemblages in the northern Neotropics. Mycol. Prog. 10:423-437. 27. Rojas, C., & Stephenson, S.L. 2008. Myxomycete APPLIED/ENVIRONMENTAL • 19 ecology along an elevation gradient on Cocos island, Costa Rica. Fungal Divers. 29:117-127. 28. Schnittler, M., Novohilov, Y.K., Carvajal, E. & Spiegel, F.W. 2013. Myxomycete diversity in the Tarim basin and eastern Tian-Shan, Xinjiang Prov., China. Fungal Divers. 59:91-108. 29. Schnittler, M., Novozhilov, Y.K., Romeralo, M., Brown, M., & Spiegel, F.W. 2012. Myxomycetes and Myxomycete-like organisms. Pp 40-88 In: Frey, W. (13th ed.) Englers Syllabus of Plant Families, Vol. 4. Bornträger, Stuttgart. 30. Schnittler, M., & Stephenson, S. L. 2000. Myxomycete biodiversity in four different forest types in Costa Rica. Mycologia 92:626-637. 31. Stephenson, S.L. 2011. Myxomycetes of the New Zealand subantarctic islands. Sydowia. 63:215-236. 32. Stephenson, S.L. 2003. Myxomycetes of New Zealand. Fungal diversity press, Hong Kong. 33. Stephenson, S.L., & Stempen, H. 1994. Myxomycetes: A Handbook of Slime Molds. Timber Press Inc., Portland, OR, USA. 34. Stephenson, S.L., Kalyanasundaram, I., & Lakhanpal, T.N. 1993. A comparative biogeographical study of myxomycetes in the mid- Appalachians of eastern North America and two regions of India. J. Biogeogr. 20:645-657. 35. Stephenson, S.L. 1989. Distribution of myxomycetes in temperate forests, II. Patterns of occurrence of bark surface of living trees, leaf litter, and dung. Mycologia 81:608-621. 36. Stephenson, S.L. 1988. Distribution and ecology of myxomycetes in temperate forests. I. Patterns of occurrence in the upland forests of Southwestern Virginia. Can. J. Bot. 66:2187-2207. 37. Takahashi, K. 2013. Myxomycete distribution varies among leaf litters of different vegetation in a local secondary forest of warm-temperate western Japan. Mycoscience 54:368-377. 38. Tran, H.T.M., Stephenson, S.L., Hyde, K.D., & Mongkolporn, O. 2008. Distribution and occurrence of myxomycetes on agricultural ground litter and forest floor litter in Thailand. Mycologia 100:181-190. 39. Unterseher, M., Schnittler, M., Dormann, C., & Sickert, A. 2008. Application of species richness estimators for the assessment of fungal diversity. FEMS Microbiol. Lett. 282:205–213. 40. Winsett, K.E., & Stephenson, S.L. 2013. Myxomycetes isolated from submerged plant material collected in the Big Thicket National Preserve, Texas. Mycosphere 4:227–231. 20 • FINE FOCUS, VOL. 1