Mycosphere List of species collected and interactive database of myxomycetes (plasmodial slime molds) for Mt. Arayat National Park, Pampanga, Philippines

Dagamac NHA1*, dela Cruz TEE1,2, Pangilinan MVB2 and Stephenson SL3

1Graduate School & Research Cluster for the Natural and Applied Sciences, University of Santo Tomas, España 1015, Manila, Philippines – [email protected], [email protected] 2Department of Biological Sciences, University of Santo Tomas, España 1015, Manila, Philippines – [email protected] 3Department of Biological Sciences, University of Arkansas, Fayetteville, Arkansas 72701, USA – [email protected]

Dagamac NHA, dela Cruz TEE, Pangilinan MVB, Stephenson SL. 2011 – List of species collected and interactive database of myxomycetes (plasmodial slime molds) for Mt. Arayat National Park, Pampanga, Philippines. Mycosphere 2(4), 449–455.

Myxomycetes (plasmodial slime molds) are fungus-like protists that are often understudied in a biodiverse country such as the Philippines. As such, the purpose of the project described herein was first to document the species of myxomycetes collected in Mt. Arayat National Park, Pampanga, Philippines and then to develop an interactive database for these organisms using DELTA software. These are the first reports of myxomycetes from Mt. Arayat. A total of 33 species were recorded for the two slopes of the mountain during the wet and dry seasons.Five species of myxomycetes (i.e., afroalpina, arcyrionema, concinnum, Enerthenema papillatum and Licea biforis) were new records for the Philippines. As a second component of the project, interactive identification guides were generated from 67 taxonomic characters using the DELTA software. The interactive database recognized spore appearance, colour of the spores by transmitted light, fruiting body colour and the outline of the sporotheca as the most useful characters in recognizing the species of myxomycetes known from Mt. Arayat. The interactive database generated in this study is the first effort along these lines for the Philippines and for Southeast Asia.

Key words – – diversity – species listing – protist – identification key

Article Information Received 21 June 2011 Accepted 12 July 2011 Published online 20 September 2011 *Corresponding author: Dagamac NHA – e-mail – [email protected]

Introduction viously by Martin & Alexopoulos (1969) for Relatively little is known of the status of the entire country. Later, Reynolds (1981) pro- myxomycetes in Southeast Asia, and this is vided a more extensive listing of Philippine particularly true for the Philippines. Most of myxomycetes, based upon published records the earlier published reports from the region and his personal collections. A total of 107 were lists of species from countries such as species were reported for the Philippines, Myanmar and Thailand (Reynolds & Alexo- including the myxomycete collections from poulos 1971). In the Philippines, Uyenco Mindanao (Davao, Cotabato and Zamboanga) (1973) reported 18 species representing 10 ge- by E.B. Copeland, and from Luzon by A.D.E. nera from 314 specimens collected from Luzon Elmer (Benguet) and E.D. Merrill (Bataan, (Quezon City and Laguna) and Mindanao Manila, Cavite and Laguna). To the best of our (Basilan and Zamboanga) during the period of knowledge, there have been no publications on 1961 to 1973. Dogma (1975) listed 46 species Philippine myxomycetes since the paper by in 20 genera, including those reported pre- Reynolds (1981), until dela Cruz et al (2009)

449 evaluated the myxomycete collection in the tain. Moist chamber cultures were prepared Mycological Herbarium of Museum of Natural from these samples. These cultures were main- History at the University of the Philippines Los tained under diffuse light at room temperature Baños, Laguna. These specimens were found to and observed regularly for the presence of be in good condition. Moreno et al (2009) myxomycetes for a period of up to eight weeks. reported a new species of Craterium, C. reti- All fruiting bodies of myxomycetes that de- sporum, from a specimen collected on Anda veloped in the moist chamber cultures were Island, Pangasinan. Macabago et al (2010) re- removed and then allowed to air-dry gradually. ported 28 species of myxomycetes belonging to Each myxomycete, together with the portion of 11 genera from La Mesa Watershed EcoPark in the substratum upon which it fruited, was Quezon City, Metro Manila. In addition, Daga placed in a small box for storage. Photographs mac et al (2010) reported five new records of of each specimen were obtained with the use of myxomycetes from the bark of Samanea sa- a Moticam 1000 (Motic, USA) camera system. mans (Jacq) Merr. collected from 21 different To identify these specimens, the fructification localities in Luzon. Herein, we report 33 spe- type (plasmodiocarp, aethaloid, sporangiate cies of myxomycetes representing 14 genera and pseudoaethaloid) and gross morphological collected from different substrata obtained in features (e.g., colouration of the peridium or Mt. Arayat National Park, Pampanga in Central stalk, presence or absence of lime, morpho- Luzon, Philippines. Moreover, we report the metrics of all vegetative structures, etc.) were first interactive database on myxomycetes de- then examined under a dissecting microscope veloped for any region of Southeast Asia. (Olympus, USA) at different magnifications. Microscopic slides also were prepared from Methods fruiting bodies to observe the presence of other The study was conducted during 2008 special characters [e.g., appearance of the ca- and 2009. All species were identified at least to pillitium, presence of a calyculus, presence of the genus level on the basis of published des- lime nodes, and spore morphology [size, shape, criptions and identification keys (Stephenson & texture, etc.)]. All slides were prepared using Stempen 1994), and also with web-based elec- 95% ethanol and lactophenol, following the tronic databases [e.g., SYNKey (Mitchell procedure described by Stephenson & Stempen 2008) and the Eumycetozoan Project (http:// (1994). Slides were then observed under a slimemold.uark.edu/)]. Taxonomic names used brightfield compound microscope (Scanner for the myxomycetes are those available from Objective to OIO, Olympus CX 31). the online nomenclatural database for the eu- mycetozoans (http://nomen.eumycetozoa.com). Development of an interactive database for myxomycetes of Mt. Arayat National Park Study Site (Fig. 1) using the DELTA software. Mt. Arayat National Park (15°12’00”N, All possible morphological characters of 120°44’31”E to 15.20°N, 120.742°E) is located the 33 species of myxomycetes from Mt. in the municipality of Arayat in the province of Arayat were initially determined under the Pampanga, on Luzon Island in the Northern stereomicroscope and compound light micro- Philippines. The park has a mid-mountain fo- scope. Each character was then assigned to any rest type with an elevation of 1,030 m above one of the five different categories using the sea level (masl). The area has an annual tempe- Character Editor of the DELTA software. rature range of 22–31°C and annual rainfall These were (1) unordered multistate: two or ranges from 284–1,844 mm. more character states that have no orderly pat- tern; (2) ordered multistate: 2 or more character Characterization and identification of collec- states that have orderly pattern, (3) integer nu- ted myxomycetes meric: any measurement in whole number (4) Samples of four different types of sub- real numeric: any measurement in fractional strata were randomly collected along the South values, and (5) text: additional information Trail (Baño) and the North Trail (Magalang), (synonyms or geographic distribution/origin of both of which extend to the peak of the moun- the specimen). Images of myxomycetes were

450 Mycosphere

Fig. 1 – Map of the municipality of Arayat in Pampanga (map source: globalpinoy.com). also included in the interactive database as a Order Stemonitales CIMAGES file to further improve the keys that were generated. The data files were then chec- Collaria arcyrionema (Rostaf.) Nann.-Bremek. ked for format errors and inconsistencies using ex Lado (site: Magalang; substratum: AL) the program CONFOR in the DELTA soft- ware. Finally, the DELTA software was run to Enerthenema papillatum (Pers.) Rostaf. (site: generate (1) an Interactive Key (INTKEY), a Baño; substratum: TW) user-friendly interactive database for species identification and species information retrieval; scintillans (Berk. & Broome) (2) Conventional Key, a dichotomous key used Morgan (sites: Baño and Magalang; substrata: for identification of species; and (3) Natural GL, BK) Language, a comprehensive description of all encoded myxomycetes. The interactive data- Stemonitis fusca Roth (site: Magalang; base can now be used for the identification of substrata: AL, GL, TW) newly collected myxomycetes. Stemonitis pallida Wingate (sites: Baño and Results and Discussion Magalang; substrata: TW, BK)

Species listing of Mt. Arayat myxomycetes Order A total of 33 species representing 14 genera are included in this annotated list of Licea biforis Morgan (site: Magalang; species from Mt. Arayat National Park. These substratum: AL) species of myxomycetes were recorded from aerial litter (AL), ground litter (GL), twigs Order (TW) and bark (BK) collected in the park. Spe- cies are arranged alphabetically under each ta- Arcyria afroalpina Rammeloo (sites: Baño and xonomic order. Examples that are new records Magalang; substratum: GL) for the Philippines are indicated in bold.

451 Arcyria cinerea (Bull.) Pers. (sites: Baño and Morgan (site: Magalang; substratum: BK) Magalang; substrata: AL, GL, TW, BK) album (Bull.) Chevall. (sites: Baño Arcyria insignis Kalchbr. & Cooke (sites: Baño and Magalang; substrata: AL, GL, TW) & Magalang; substratum: TW) Physarum bivalve Pers. (sites: Baño & Arcyria pomiformis (Leers) Rostaf. (site: Magalang; substrata: AL,TW) Magalang; substratum: TW) (Batsch) Pers. (site: Baño; Perichaena chrysosperma (Curr.) Lister (site: substrata: AL, GL) Baño; substratum: GL) Phyarum compressum Alb. & Schwein. (site: Perichaena depressa Lib. (site: Baño; Baño and Magalang; substrata: AL, GL, TW) substratum: GL) Physarum echinosporum Lister (site: Baño; Order Physarales substrata: AL, GL)

Craterium concinnum Rex (site: Baño; Physarum leucophaeum Fr. (sites: Baño and substratum: AL) Magalang; substrata: AL, GL)

Cribraria microcarpa (Schrad.) Pers. (site: Physarum pusillum (Berk. & M.A. Curtis) G. Baño; substratum: BK) Lister (site: Magalang; substrata: GL, TW)

Diachea bulbillosa (Berk. & Broome) Lister Physarum roseum Berk. & Broome (site: (site: Baño; substratum: BK) Magalang; substratum: TW)

Diachea leucopodia (Bull.) Rostaf. (site: Physarum viride (Bull.) Pers. (site: Baño; Magalang; substratum: AL) substrata: GL, TW)

Diderma effusum (Schwein.) Morgan (sites: These are the first reports of myxomy- Baño and Magalang; substrata: AL, GL) cetes from Mt. Arayat National Park. Interes- tingly, five of the species listed herein are new hemisphaericum (Bull.) Hornem. records for the Philippines. These are Arcyria (sites: Baño and Magalang; substrata: AL, GL) afroalpina, Collaria arcyrionema, Craterium concinnum, Enerthenema pappilatum and Diderma subasteroides M.L. Farr (site: Baño Licea biforis. The last comprehensive listing of and Magalang; substrata: BK, GL, TW) Philippine myxomycetes was published by Reynolds (1981). He presented an annotated iridis (Ditmar) Fr. (sites: Baño and list of 107 species based on published and un- Magalang; substrata: AL, GL, TW) published records. Of these, 53 species were listed as new records for the country, and these Didymium nigripes (Link) Fr. (site: Magalang; were members of the genera Arcyria, Badha- substratum: GL) mia, , Clastoderma, Comatricha, Craterium, Cribaria, Diachea, Dictydium, Didymium ochroideum G. Lister (site: Baño; Diderma, Didymium, , , substratum: GL) Hemitrichia, Lamproderma, Licea, , Metatrichia, Perichaena, Physarella, Physa- Didymium squamulosum (Alb. & Schwein.) Fr. rum, Stemonitis, Trichia and Tubifera. Most of (sites: Baño and Magalang; substratum: AL, these myxomycetes were collected from sub- GL) strata obtained from several sites in the coun- try, including Luzon, Visayas and Mindanao. Physarella oblonga (Berk. & M.A. Curtis) The list compiled by Reynolds corresponded to

452 Mycosphere 25% of all known species at the time it was ters were those recommended initial characters published and represented approximately 60% that needed to be checked prior to the of the estimated total Philippine myxomycete application of the interactive database. The flora. Recently, Corpuz et al (2009) also INTKEY program acts as a functional diagnos- studied the myxomycetes flora in selected tic tool for the rapid identification of specimens highland and island localities in Luzon. About and for information retrieval (Dallwitz et al. 25 species were observed in highland localities 2000). The INTKEY displays information on in Luzon, while approximately 30 species were the taxonomy of the species, including their recorded from different islands in Pangasinan. illustrations and other relevant data. The Interestingly, four new records for the Philip- program also describes diagnoses, summarizes, pines, Elaeomyxa miyazakiensis, and finds similarities and/or differences tigrinum, Perichaena pedata and Physarum between members of related taxa (Dallwitz et decipiens were noted. Moreover, a specimen al. 1995, 2000). found on Anda Island, Pangasinan was identi- Another identification guide generated fied as a species of Craterium new to science, with the DELTA software was a conventional which was described as C. retisporum by or dichotomous key. The dichotomous key Moreno et al (2009). Dagamac et al (2010) also categorizes the species based on important collected five new records for the Philippines. characters. These characters can be used to These were Clastoderma microcarpum, Dia- distinguish unknown species from other nema harveyi, Diderma subasteroides, Phy- species in the database. In the present study, 49 sarum leucophaeum and Stemonitis pallida. All of the 67 encoded characters were found to be five were obtained from bark samples of useful in generating the dichotomous key. On Samane samans (Jacq) Merr. collected from 21 the practical side, the generated dichotomous different localities in Luzon. This now brings key can be printed and used as field guide. The the total number of myxomycetes recorded Natural Language was another identification from the Philippines to 122. However, the guide generated by the DELTA software. The reported number is still considered small for a Natural Language listed all information for a tropical country. This suggests that there are particular species encoded in the database. still many more species remaining to be found Taxonomic descriptions written in the Natural in less studied and relatively unexplored parts Language format include synonyms, fruiting of the Philippines. body descriptions, vegetative parts and spore morphologies. Since descriptions of all taxa in Interactive database for Mt. Arayat myxo- the database cannot be generated using the mycetes using the DELTA software. INTKEY or the Dichotomous Key, the Natural A total of 67 characters were encoded Language provides descriptions of all previous- using the DELTA software program. Forty- ly encoded characters. nine were unordered multistate characters, ten The DELTA software has already been were text characters, while six were real applied successfully to myxomycetes in the numeric characters. Thirty-eight of the 67 Order Echinosteliales (Pando 1999) and to characters were independent characters, thus several groups of plants (Alejandro & Liede allowing the program to infer that certain 2003, Watson & Dallwitz 2004, Bean 2005, characters are applicable only to certain taxa. Watson & Dallwitz 2005, Alejandro & Meve From the 67 encoded characters, only 55 were 2008). The interactive database generated for found useful in differentiating the different the myxomycetes of Mt. Arayat is user-friendly myxomycete taxa from Mt. Arayat, and thus and contains information on the taxonomy of were useful in generating INTKEY (Figure these organisms and other relevant information 2b). Characters such as spore appearance, color (e.g., their biogeography and/or economic of the spores by transmitted light, fruiting body importance). Furthermore, this database can be colour and the outline of the sporotheca were updated regularly and requires minimal main- generally the most useful characters in tenance. The DELTA interactive database recognizing species, and thus were scored with generated in this project was the first for myxo- high reliabilities. Consequently, these charac- mycetes from the Philippines, or anywhere

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Fig. 2 (a–d) – Features of the interactive database for Mt. Arayat myxomycetes as generated from the DELTA software a DELTA editor b Interactive Key c Conventional key d Natural language. else in Southeast Asia. This online database Research and Development (PCHRD) for the can help other myxomycologists to easily share graduate scholarship and thesis grant. The new discoveries relating to myxomycetes, most authors thank the Studienstiftung Mykologie specifically the global distribution patterns of through Prof. Walter Gams for the research these organisms. It also helps other researchers grant needed to complete this study and Mr. to overcome some of the difficulties in iden- Rogelio T. Trinidad and Mr. Regidor M. de tifying myxomycetes to species. Since this can Leon, Department of Environment and Natural be updated at any time, data specifications Resources (DENR) Region III for the gratui- (e.g., characters and its character states for each tous permit to collect specimens. We also thank taxa) that were introduced in the development Mr. Paul Richard J. Yulo and Mr. Benedict A. of the database can be shared and further Maralit for technical assistance during the field developed among researchers interested in the collection and database development. study of myxomycetes. Through the use of the DELTA software, conventional identification References keys and interactive keys can be easily gene- rated and can be valuable for the taxonomist Alejandro GJD, Meve U. 2008 – Development working with myxomycetes. of an interactive database to the species of Philippine Mussaenda (Rubiaceae). Acknowledgements Philippine Journal of Systematic Biology NHAD would like to acknowledge the 2, 12–16. Department of Science and Technology Alejandro GD, Liede S. 2003 – The Philippine (DOST) Philippine Council for Health Rubiaceae genera: updated synopsis in

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