unesco man and the biosphere programme · 2016-04-19 · unesco man and the biosphere programme...

UNESCO Man and the Biosphere Programme

Young Scientists Award 2014

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Final Report

Biodiversity, taxonomy, ecological patterns and conservation of myxomycetes and

macrofungi in Puerto Galera Biosphere Reserve and Sablayan Watershed Forest

Reserve, Mindoro, Philippines

By

Dr. Thomas Edison E. dela Cruz

Research Center for the Natural and Applied Sciences

University of Santo Tomas

Manila, Philippines

2014 MAB Young Scientist Award

October 2015



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I. Executive Summary

The Island of Mindoro in the Philippines is known for its

wide variety of endemic species of plants and animals. It

is likewise recognized as one of the most naturally

diverse islands in the country. UNESCO also recognized

the Puerto Galera Peninsula including Mt. Malasimbo as

part of its Man and the Biosphere programme. Equally

important for biodiversity is the lowland forest in Mt. Siburan in Sablayan Watershed Forest Reserve,

also in the Island of Mindoro. However, the island has only a few remaining forest ecosystem. There

is also limited studies conducted on its microflora. To better assess the island’s biodiversity, it is

important to document all species present in its remaining forest ecosystems, including the less-

documented slime molds (myxomycetes) and macrofungi. In this research study, we surveyed several

collecting points within the lowland forest ecosystems in Mt. Malasimbo in Puerto Galera, Oriental

Mindoro and in Mt. Siburan in Sablayan, Occidental Mindoro during two field collection trips,

October 2014 and June 2015. Identification of the collected specimens were done either by

morphological characterization or combined morphological and molecular methods. Results of the

study identified a total of 48 species of myxomycetes, a high number of records comparable to recent

myxomycete studies in the Philippines. More species of slime molds were recorded in Mt. Siburan

than in Mt. Malasimbo. A higher species diversity was therefore computed for Mt. Siburan. A low

CC value between the two study sites indicate dissimilarities in their species composition. Between

the different substrate types, decayed woody vines and twigs harbored the most diverse species of

myxomycetes. Among the macrofungi collected in the area, a total of 34 species belonging to 21

genera and 13 families were recorded. Molecular methods confirmed the identities of the collected

macrofungi with high bootstrap supports. A higher number of species of macrofungi were again

reported for Mt. Siburan than in Mt. Malasimbo. The research study further provides baseline

information for the profiles of myxomycetes and macrofungi in a local scale of Mindoro Island and

will also contribute to the understanding of their distribution in the whole of the Philippines and in

the tropical ecoregions.

Keywords: biosphere reserve, fungi, fungus-like protists, lowland forests, species diversity



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II. Introduction

Entirely surrounded by tropical seas, the Philippine Islands are isolated from the Asian

landmass by hundreds of kilometers of open water. Such geographic isolation coupled with its

tropical climate has resulted in a high level of endemism (up to 85%) among its flora and fauna and

probably also among its microbial biota. However, the country is also listed as one of the most

threatened ecosystems on the planet. Only about 7% of the total land area is covered by primary

forests. Thus, there is a great urgency to document the country’s vanishing biodiversity.

The Island of Mindoro is recognized as one of the most naturally diverse areas in the

Philippines. It is also known for its profound sceneries. In fact, Puerto Galera in Oriental Mindoro is

considered as one of the country’s top tourist destinations, owing to its beautiful beaches with

unblemished water and other recreational activities. Puerto Galera is located 130 km from the capital

city of Manila. Towering above Puerto Galera are Mt. Malasimbo (1,228 meter above sea level) and

Mt. Talipanan (1,185 masl) which contain one of the dense coastline rainforests in the island. In 1973,

UNESCO declared the peninsula as part of the Man and the Biosphere programme due to the less

successful protection of these terrestrial areas as a result of an increase in human presence. In fact,

encroachment and conversion of mountain slopes to farmlands and establishment of many beachfront

resorts along the coastline are now evident. This, in turn, makes Puerto Galera and its mountains as

one of the most threatened environments in the Philippines. The Sablayan Watershed Forest Reserve

is another lowland rainforest in the west coast of Occidental Mindoro. The rainforest in Mt. Siburan

in this watershed is also showing signs of disturbance by human resettlements and therefore, also

represents an important area for protection of Mindoro wildlife along the west coast. The watershed

is also gaining attention as another ecotourism site in the region. The influx of ecotourists into these

biosphere reserves, although it will definitely boost the local economy, will undoubtedly have a

significant impact on nature in the area. Before any of the species present—plants, animals, fungi and

myxomycetes become influenced by man-made activities, it is of urgent importance to assess

Mindoro Island’s biodiversity, particularly the relatively poorly documented groups such as

myxomycetes and macrofungi that are virtually unknown in this part of the country.

Myxomycetes or myxogastrids are cryptogamic protists that are widely dispersed in terrestrial

habitats. These organisms are also referred to as plasmodial slime molds and were previously

categorized as Fungi (Adl et al., 2005; Baldauf, 2008) but later classified as Protoctista because of

the amoeboid stage in its lifestyle (Spiegel et al., 2004; Pawlowski & Burki, 2009). Molecular studies

also supported this classification (Baldauf & Doolittle, 1997; Baldauf et al., 2000). Hence, this

unusual characteristic makes them a model organism to study biological processes, particularly

physiology and cell differentiation (Everhart & Keller, 2008). Myxomycetes are widely distributed

both in temperate and tropical regions. Studies showed the presence of myxomycetes in Guatemala

and Costa Rica, (Rojas et al., 2012; Schnittler & Stephenson, 2002), Thailand (Tran et al., 2008; Ko



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Ko et al., 2010), Mexico (Estrada-Torres et al., 2009), USA (Ndiritu et al., 2009), South America

and Chile (Wrigley de Basanta et al., 2010, 2011), Singapore (Rosing et al. 2011), Myanmar (Ko Ko

et al.. 2013), and Laos (Ko Ko et al., 2012). They have been reported in varied substrata, e.g. in

coarse woody debris on the forest floor, the bark surface of living trees, forest floor litter, the dung

of herbivorous animals, soil, and aerial liter (Stephenson 1988, 1989; Stephenson & Stempen, 1994).

The very first record of myxomycetes in the Philippines was made by Elmer and Merrill in

early 1900s and later by Uyenco (1973), Dogma (1975) and Reynolds (1981). A total of 107 species

was listed and accounted for the Philippines during this time (Reynolds, 1981). Recently, researches

on myxomycete diversity and distribution in the Philippines were reported in the Bicol Peninsula and

Quezon Province (Dagamac et al., 2015a, 2015b), Mt. Maculot in Batangas (Cheng et al., 2013), Mt.

Arayat in Pampanga (Dagamac et al., 2011), La Mesa Ecopark in Quezon City (Macabago et al.,

2010), Lubang Island in Occidental Mindoro (Macabago et al., 2011) and Anda Island in Pangasinan

(Kuhn et al., 2013). To date, about 150 species of myxomycetes are accounted for the Philippines

based on published and unpublished reports. However, despite the previous effort on myxomycete

studies, still this number of species is relatively low for a tropical country like the Philippines.

Another group of organisms commonly found in forest ecosystems are the macrofungi

belonging to the Division Ascomycota and Basidiomycota of the Kingdom Fungi. For many of these

fungi, they form symbiotic relationships with plants and animals (Claridge et al., 1996). They are

also important in ecosystems as they are involved in the decomposition process that allows recycling

of nutrients. Macrofungi such as mushrooms are also known to have a broad range of uses as food

and medicine (Chang & Miles, 1987; 2004). In fact, both wild and cultivated mushrooms have been

known for their nutritional and culinary values. In addition to their nutritional value, certain

mushrooms are also abundant sources of a wide range of useful natural products. For example,

various compounds including terpenoids, steroids, phenols, and alkaloids, which have been isolated

and identified from the fruiting body, culture medium, and culture broth of mushrooms, were shown

to have promising biological effects, preventing a range of diseases such as hypertension,

hypercholesterolemia, diabetes and cancer (Lindequist et al., 2005). These are but some of the uses

of macrofungi.

In spite of their economic importance and ecological role, few studies on macrofungal

diversity have been conducted in the Philippines. Earlier study of Quimio and Capilit (1981) noted

that the Philippines has 3,755 fungal species. Recently, the study of Tadiosa et al. (2011) in the Bazal-

Baubo Watershed in Aurora Province also denoted high macrofungal species diversity in this area.

Some of the fungal species collected here were Agaricus sp., Lepiota aspera, Lepiota cristata,

Amanita fulva, Auricularia auricula, Auricularia delicata, Panaeolus sp., Strobilomyces

strobilaceus, Cantharellus sp., and Coprinus disseminates. The fungal species Aseroe rubra Labill.,

Lycoperdon echinatum Pers., Macrolepiota rhacoes (Vittadini) Singer, and Cookeina tricholoma

(Mont.) Kuntze were also first recorded in Aurora. The Taal Volcano Protected Landscape in Talisay,

Batangas was also reported to have vast macrofungal communities. Macrofungi under the families



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Ganodermataceae, Polyporaceae, Auriculariaceae, and Xylariaceae occurred in forested and open

areas in Taal. A small number of rare species were also identified such as Cookeina sulcipes, Galiella

rufei, Dictyophora duplicate, Cymatoderma elegans, Microporus vernicipes, and Xylaria longipes

(Tadiosa & Briones, 2013). Macrofungi were also described in Mt. Palay-Palay National Park in

Cavite (Tadiosa et al., 2005). De Leon et al. (2013) likewise identified several macrofungi in six Aeta

communities in Central Luzon. These studies indicate high species richness for mycoflora, but still

many areas remained unexplored including the Island of Mindoro.

This research project therefore is directed towards the goal of documenting the myxomycetes

associated with leaf litter and dead twigs in Mt. Malasimbo in the Puerto Galera Biosphere Reserve

in Oriental Mindoro. The project also examined the macrofungi associated with forest floor leaf litter

and decayed logs in the forest ecosystem on the Puerto Galera peninsula. The study also looked at

another forest ecosystem, Mt. Siburan in Sablayan Watershed Forest Reserve in Occidental Mindoro

as another study site for comparison within the island of Mindoro. The similarities on the types of

vegetation between Puerto Galera and Sablayan make it as environmentally suitable for comparative

diversity analysis. It is a worthwhile endeavor to compare the assemblages of myxomycetes and

macrofungi between these lowland mountain forests as more evidence of human activities are noted

in Mt. Malasimbo than in Mt. Siburan, thereby, providing additional evidence for possible effects of

human activities to species biodiversity as exemplified by myxomycetes and macrofungi in this study.

III. Materials and Methods

A. Study sites and collecting localities

Puerto Galera is bounded on the north by Verde Island passage, which separates it from the

mainland of Luzon. The mainland (120’50’ to 121’60’ E; 13’20’ to 13”25’ N) lies on the northern

part of Mindoro Island, about 130 km south of Manila. On the other hand, Sablayan Watershed Forest

Reserve (120o 55.00' E; 12o 48.00' N) is located in the southwestern of Mindoro (BirdLife

International, 2015).

Site 1. Mt. Malasimbo. This forest is mid-elevated (1,228 meter above sea level) containing dense

coastline dipterocarp rainforest. The mountain slopes are steep.

Site 2. Mt. Siburan. The Siburan forest is located within Sablayan Prison and Penal Farm of Sablayan

Watershed Forest Reserve. It is a prime spot where endangered species of pigeons and endemic

Tamaraw (Bubalus mindorensis) could be found. This pristine forest surrounds the picturesque 24-

hectare inland Libuao Lake and has an open forest floor. Furthermore, it is considered to be the largest

tract of lowland forest known in Mindoro (BirdLife International, 2015).Generally, it has a closed

canopy with trees of up to 25 meters or more.



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Three to five collecting localities were identified for each study site. Collection was done during the

month of October (2014). An additional field collection was done during the month of June (2015).

Figure 1. Map of study sites in Puerto Galera, Oriental Mindoro (Mt. Malasimbo) and Sablayan

Watershed Forest Reserve, Occidental Mindoro (Mt. Siburan) (Quantum GIS, v1.8).

Climatological data. Mindoro Island is characterized by two types of climate. The Oriental Mindoro

has Type III with no very pronounced seasons while Occidental Mindoro has Type 1 with two

pronounced seasons. However, both areas are experiencing almost dry season from November to

April and wet during the rest of the year. It received an average rainfall of 2,059.9 mm in which the

month of November had the highest rainfall with 450.2 mm. The month with the lowest rainfall is

February with 2.6 mm. The annual prevailing wind direction is northeast. Oriental Mindoro is more

often directly affected by tropical cyclones during the latter part of the typhoon season: October and

November. Because of its protected topography, maximum winds could be much less than that

observed in the surrounding areas. The coldest months were December and January in which

minimum temperatures were near 20 ºC. The relative humidity of the area was about 80% (PAG-

ASA, DOST).

Mt. Malasimbo, San Isidro, Puerto Galera

Sablayan Watershed, Malisbong, Sablayan

13°28’30”N, 120°54’59”E

13°28’19”N, 120°54’41”E

12°49’40.7”N, 120°54’57.1”E

12°49’2.3”N, 120°53’48.5”E



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Mt. Malasimbo, Puerto Galera Mt. Siburan, Sablayan

Figure 2. The lowland mountain forests in Mt. Malasimbo, Puerto Galera and Mt. Siburan, Sablayan

in Mindoro Island, Philippines.

Figure 3. The expedition team during the field collection on October (2014, left) and June (2015,

right).



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B. Myxomycetes

B.1. Collection of field specimens and substrata

Fruiting bodies of myxomycetes observed in the field were collected and recorded following

the methods described by Stephenson (1988). All specimens were directly glued inside the herbarium

boxes for permanent storage. In addition to the field collections, different types of substrata were also

collected. For each sampling point or locality in Mt. Malasimbo and Mt. Siburan forests, aerial leaf

litter (AL), aerial woody vines (WV), ground leaf litter (GL) and twigs (TW) were collected. As

previously described (Stephenson & Stempen, 1994), AL and WV refer to dead but still attached and

had not been in contact with the ground while TW and GL have already been dropped on the forest

floor. Ten samples for each substratum in each sampling point were randomly collected and placed

in a medium-sized, clean brown paper bag with the proper label of study sites, type of substrata and

date of collection. The samples were then brought to the laboratory and air-dried for seven to ten days

if the samples are wet. This was done prior to the preparation of moist chambers (MC).

B.2. Preparation of moist chambers

The use of moist chamber has been widely known for myxomycete cultivation (Harkonen,

1981; Lado et al., 2003; Wrigley de Basanta et al., 2008; Kilgore et al., 2009), and reported to be

excellent technique to assess the diversity of myxomycetes in a particular habitat type or study site

(Novozhilov et al., 2000). Therefore, in this study, moist chamber technique were also performed for

each collected substrata as described by Stephenson & Stempen (1994). For the aerial (AL) and

ground (GL) leaf litter, this were cut into postage-stamp sized pieces and 8-10 pieces of the cut leaves

were placed in disposable petri dishes (90mm) lined with filter paper. Similarly, twigs and woody

vines were cut into 2-3 inches length and approximately five pieces were placed in petri dish. Each

moist chamber culture were then dispensed with distilled water until all the materials were completely

submerged. Following incubation for 24 hours, the pH of each culture were determined by placing

the electrode of the pH meter (Sartorious PB-11) on the substrata soaked in distilled water. After

getting the pH, the excess water were poured off from the samples and the moist chambers were

incubated at room temperature under diffuse light. All moist chamber set-ups were examined at least

once a week for over a period of eight to ten weeks. The moisture of each moist chamber were

maintained by adding small amounts of water occasionally during the observation period (Stephenson

& Stempen, 1994; Rojas et al., 2012). The moist chambers were checked for the presence of

myxomycete plasmodia and/or fruiting bodies. When fruiting bodies of a given species develop more

than once in the same culture, it was considered as one single collection. However, moist chamber

were recorded as negative if no fruiting bodies and/or plasmodia or slime track was observed. Only

those petri dishes that yielded fruiting body and/or plasmodial growth were noted as positive for

myxomycetes.



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B.3. Preparation of voucher specimen and characterization and identification of myxomycetes

As soon as the fruiting bodies fully mature, the portion of the substrate upon which the fruiting

body occurred were removed from the moist chamber culture, allowed to dry and then glued in a

small paper box suitable for long-term storage (Keller & Braun, 1999). All collected specimens were

deposited at the herbarium collection and inputted in the database record of the Pure and Applied

Microbiology Laboratory, Research Center for the Natural and Applied Sciences, University of Santo

Tomas in Manila, Philippines.

Identification of each myxomycete species were based solely on the overall morphological

characteristics of its identifiable fruiting body (Lado, 2001). These included the type and size of

fruiting body, shape of sporotheca, appearance of the stalk and presence of lime (both on sporotheca

and stalk) (nomenclatural protocols of Lado, 2005-2011). Moreover, spores of each species were

examined with a compound microscope (Olympus CX3112C04). To do this, fruiting body of

specimen were mounted on a slide with the use of potassium hydroxide (KOH) and/or lactophenol as

a mounting medium as described in detail by Keller et al. (2008). Identifications of myxomycete

collections was done at least up to the genus or species level following comparison with published

literatures and identification keys, e.g. Stephenson & Stempen (1994), and also with web based

electronic databases, e.g. SYNKey (Mitchell, 2008) and the Eumycetozoan Project

(http://slimemold.uark.edu/). To further validate the identification using the observed characteristics,

an online nomenclatural database for the eumycetozoans (http://nomen.eumycetozoa.com) were also

used.

B.4. Diversity and ecological analysis

Percent Yield. The productivity of the moist chambers (MC) in the substrate and study sites were

separately computed. As previously described by Stephenson (1988), the presence of plasmodia or

fruiting body in each of the MC were counted and considered as one collection. To compute this, the

total positive collections were divided by the total number of MC prepared multiplied by 100.

Species Composition and Occurrence. The occurrence of each species in each substrate type and

study site were determined based on its relative abundance (RA). To do this, the presence and absence

of fruiting bodies of myxomycetes from the moist chambers were checked and counted. To compute

for the RA, the total number of myxomycete records in a specific substrate/study site were divided

by the total number of collections. The RA index of each species were then determined by placing it

in categories following the modified ranking of Rojas et al. (2012) in which each species were

regarded as: (1) abundant (A) if the RA value is equal or more than 3 % of total number of collections,

(2) common (C) if the RA value is falling between 1.5 % and 3 % of total number of collections, (3)

occasional (O) if the RA value is in between 1.5 % and 0.5 %, and (4) rare (R) if the RA value is less

http://nomen.eumycetozoa.com)/



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than 0.5%. Furthermore, species listing along with its RA, substrate type and collecting point were

included in this report.

Taxonomic Diversity. Taxonomic diversity was computed by getting the ratio of the number of

species with the number of genera (S/G ratio). A value for the S/G ratio is inversely proportional to

its taxonomic diversity; thus, the lower the S/G ratio the more diverse a particular biota is considered.

Stephenson et al. (1993) noted that a biota in which the species are divided among many genera are

more diverse in a taxonomic sense than one in which most species belong to only a few genera.

Species Diversity. In order to assess and quantify the myxomycetes diversity, species diversity indices

were calculated between substrates types and study sites as described by Stephenson (1989) and

Dagamac et al. (2012). This were computed as follows:

i. Shannon Index of Diversity (HS)

where R = species count

pi = proportion of R represented by the ith species

ii. Gleason Index of Species Richness (HG)

where Np= total number of species

Ni= total number of individuals in the ith species

iii. Pielou’s Index of Species Evenness (E)

where HS= Shannon Index of Diversity

Hmax= the maximum value of HS

Equation 1:

− ∑ i (pilnpi) HS =

R

i=1

Equation 2:

lnNi

lnNi

HG =

Equation 3:

E = Hs

Hmax

Np-1

lnNi



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Community Analysis. To test the similarities of the communities of myxomycetes in the study areas

and substrates, coefficient of community (CC) and the percentage similarity (PS) indices were

calculated as described by Stephenson (1989). In the coefficient of community, the presence or

absence of species were considered. A CC value close to one (1.0) indicates that both communities

have the presence of all species of myxomycetes and zero (0) when no species was present in both

communities to be compared. The PS index were computed in which the relative abundance of species

and not only their presence are considered. The PS values ranges from 0 to 1. A higher PS value

indicates that the two communities being compared are more similar in terms of species composition

and abundance (Dagamac et al., 2010).

C. Macrofungi

C.1. Collection of field specimens

In this research study, all visible macrofungi on soil, dead woods (i.e., logs, barks or twigs)

and leaf litter encountered within the sampling points were collected randomly and placed on a

collection basket together with their substrate. A knife were used to remove the specimens from their

substrates. The woody macrofungi were placed in a collection basket while the fleshy macrofungi

were initially stored in separate air-tight vials or bottles to prevent deterioration of the specimens.

Photos of the specimens in their natural habitat were also taken. All collected specimens were then

transported to the camp site for preliminary processing before being transported to the laboratory.

C.2. Preservation and preparation of herbarium specimens

All collected macrofungi were preserved or prepared as herbarium specimens. For bracket

and woody macrofungi, these were dried inside a fruit drier at a temperature of 40-50°C for 24-48

hours or until completely dried. After drying, the specimens were then placed in herbarium boxes and

kept in a Ziploc plastic bag with silica gel to prevent moisture and mold formation. For fleshy and

jelly macrofungi, specimens were placed in small plastic containers or vials with 70% ethanol for

preservation. All macrofungal specimens were labeled with the specimen code, date and place of

collection, and substrate. The collected macrofungi were deposited at the Pure and Applied

Microbiology Laboratory, Research Center of the Applied and Natural Sciences, University of Santo

Tomas in Manila, Philippines.

C.3. Characterization and identification of macrofungi

Morphological characterization. Preliminary identification of the macrofungi were based on their

morphological characters. Detailed descriptions of each of the collected macrofungi were made on

previously prepared macrofungi identification sheet. Morphological characteristics that were

recorded were as follows: substrate type, description of pileus (diameter, shape, apex, surface, color,

peeling, and margin), description of lamellae (gills, attachment, arrangement) and description of stipe

(color, height, width, shape, attachment to cap, surface, annulus, attachment to substrate and volva)



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following modifications of the data sheets developed by Leonard (2010). These were mainly used for

members of the Agarics and other related groups. Additional characters were also noted for other

species.

Molecular Identification. To confirm the identities of the macrofungi, genomic DNA of selected

macrofungi were extracted following a modified CTAB protocol and then sequenced. To do this, 500

µl of CTAB (2% Tris-HCl with pH 8.0, 100mM EDTA with pH 8.0, 20mM NaCl, 1.4M CTAB) was

pipetted to sterilized Eppendorf tubes. About 200 mg of the dried fungal samples were placed in the

Eppendorf tubes using forceps. The samples in the tubes were then crushed and ground using a small

pestle and placed in an incubator at 65°C for 30 minutes. A combination of 500 µl

Phenol:Chloroform:Isoamyl alcohol (PCI) was then added to the samples. The tubes were vortexed

afterwards and placed in a centrifuge for 15 minutes at 12,000 rpm, 4°C. After this step, there is a

clear distinction of layers in the Eppendorf tubes. Only 150 µl x 2 of the upper clean layer was placed

in a second batch of Eppendorf tubes. The remaining layer/s of the sample was discarded. Then,

210µl Isopropyl alcohol were transferred to each tube. The tubes were then moved gently sideways

to mix the liquid layers. After which, the samples were placed in a rack for 10 minutes at room

temperature and then, centrifuged for 15 minutes at 12,000 rpm, 4°C to get the pellet. The liquid

portion was then poured out and the tubes were placed in a rack to dry. Ice cold 70% ethanol (500 µl)

was then added to each tube and the tubes were then shaken gently. The Eppendorf tubes were again

centrifuged for 5 minutes at 12,000 rpm, 4°C to get the pellet. The remaining liquid was removed

and the Eppendorf tubes were placed in paper towel to dry. To finish, 500 µl of distilled water was

added to the tubes and incubated for 20 minutes at 65°C. The samples were then stored in a

refrigerator until use. For PCR amplification and gene sequencing, the forward ITS1 (5'-

TCCGTAGGTGAACCTGCGG-3') and reverse ITS4 (5'-TCCTCCGCTTATTGATATGC-3')

primers were used to amplify the rDNA ITS region. The PCR amplification parameters were as

follows: initial denaturation at 95°C for 3 minutes (initial separation of DNA strands), 35 cycles at

95°C for 40 seconds (separation of DNA strands), 55°C for 40 seconds, and 72°C for 1 minute, and

final extension at 72°C for 10 minutes (final synthesis of DNA). To check for the PCR products, the

PCR products (3 µl) and loading buffer (1 µl) were loaded in 1% agarose gel mixed with 0.5x-1x

TAE buffer. Gel electrophoresis was set up at 110V for 25 minutes and the DNA bands were

visualized using Bio-Rad imager machine. Purification of the PCR products was done using the

commercially available kit GeneAll Biotechnology (South Korea). After which, all PCR products

were sent for sequencing at Macrogen Inc., Seoul, South Korea. The sequences were uploaded to

NCBI BLAST to determine closely related sequences. Based on the results of the BLAST search and

the previous studies on the genus of the macrofungi, sequences were selected for alignment using

MEGA 5.2 software. Maximum likelihood trees were also constructed using MEGA 5.2.

Phylogenetic trees were constructed to show the relatedness of the collected macrofungal species.



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C.4. Taxonomic diversity

In this research, all macrofungal species were listed alphabetically under their respective class

and family. Furthermore, the number of species and genera for each study sites or collection period

was also computed, recorded and computed.

IV. Major Findings

A. Myxomycetes of Mt. Siburan and Mt. Malasimbo, Mindoro Island, Philippines

Myxomycetes or slime molds play a very important role in terrestrial forest ecosystems. They

have been identified from varied substrata, e.g. coarse woody debris (Stephenson, 1988), ground litter

(Tran et al., 2008), aerial litter (Rojas & Stephenson, 2008), bark of living trees (Wrigley de Basanta,

2000), soil (Stephenson et al., 2004a), dung (Stephenson, 1989), inflorescences (Schnittler &

Stephenson, 2002), lianas (Wrigley de Basanta et al., 2008), and decaying fronds (Stephenson, 2003).

About 1,000 species are so far described worldwide. In the Philippines, recent studies updated the

number to 150. In this study, a total of 26 records of myxomycetes were noted from field collections

in the two forest sites in Mindoro Island (Table 1). Ten species were observed from Mt. Siburan

while a slightly higher number, 12 species, were noted for Mt. Malasimbo. These species were

collected from different substrata ranging from decayed leaf litter, woods or logs, and fruits. The

specimens were collected during the two field surveys on October 2014 and June 2015. During these

field collections, substrata were also collected and set-up in moist chamber cultures.

A total of 1,260 moist chambers (540 for Mt. Malasimbo, 720 for Mt. Siburan) were set-up

from the collected substrata, e.g. aerial (AL) and ground (GL) leaf litter, woody vines (WV), and

twigs (TW). A higher productivity (80%) was observed from samples collected in Mt. Malasimbo

than in Mt. Siburan (Table 2). Among the collected substrata, the highest productivity (87%) was

recorded for AL collected in Mt. Malasimbo followed by TW from the same mountain site.

Interestingly, highest productivity was also observed for AL among the substrata collected in Mt.

Siburan. Aerial leaf litter are good spore traps and could account for its higher moist chamber

productivity. Ground leaf litter was the least productive of all the collected substrata for both study

sites.



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Table 1. List of field collections of myxomycetes from the lowland forests in Mindoro Island.

Mt. Siburan Substrates Number of Records

Ceratiomyxa fruticulosa var

arbuscola

decaying bamboo

1

Didymium nigripes dried fruit still attached to the plant 1

Didymium squamulosum ground litter 2 Cribraria cancellata twigs 1

Cribraria microcarpa decayed wood 1

Perichaena chrysosperma twigs 1

Didymium nigripes ground litter 1

Arcyria denudate decaying bamboo 2

Stemonitis fusca decayed wood 1

Physarum stellatum decayed wood 1

Mt. Malasimbo

Stemonitis sp. decayed wood 1

Hemitrichia serpula decayed wood 1

Physarum leucophaeum bark 1

Diachea leucopodia ground litter 2

Didymium iridis ground litter 1

Craeterium leucophaeum ground litter 2

Arcyria denudata decayed wood 1

Arcyria cinerea decayed wood 2

Didymium squamulosum ground litter 1

Hemitrichia calyculata twigs 1

Physarum viride decayed wood 1



page. 15

Table 2. Productivity of moist chambers.

STUDY SITES

PLASMODIUM FRUITING BODY MYXOMYCETES

Mt. Siburan 17% 49% 65%

Mt. Malasimbo 32% 48% 80%

SUBSTRATE TYPES

Mt. Siburan PLASMODIUM FRUITING BODY MYXOMYCETES

AL 24% 55% 79%

GL 14% 36% 50%

TW 19% 49% 68%

WV 8% 66% 73%

Mt. Malasimbo PLASMODIUM FRUITING BODY MYXOMYCETES

AL 32% 55% 87%

GL 3% 30% 66%

TW 27% 59% 86%

In this study, a total of 48 species of myxomycetes were so far recorded for the two forest

sites. The number were comparable to the number of species recorded in Mt. Arayat in Pampanga

(Dagamac et al., 2012; Dagamac et al., 2014), in Mt. Makulot in Batangas (Cheng et al., 2013), and

in Mt. Kanlaon in Negros Oriental (Alfaro et al., 2015). Highest number of collections were recorded

for Arcyria cinerea, Diderma hemisphaericum, D. effusum, Lamproderma scintillans, Physarum

melleum, Perichaena pedata, and Stemonitis sp. (Fig. 4). Differences between the records of

collections per species were noted for the two mountain sites.

All fruiting bodies of myxomycetes were characterized under a stereomicroscope and a

compound light microscope. The detailed descriptions of the size, color, shape, and appearance of

fruiting bodies, the size, color, shape and texture of the spores, and the presence of unique features

such as capillitium and lime nodes (calcium carbonate) were used to identify the collected species.

Figures 5-6 showed representative species of myxomycetes collected and identified in the two forest

sites in Mindoro Island. Some of the species were also subjected to scanning electron microscopy for

detailed study on their fruiting body and spore morphologies (Fig. 7).



page. 16

Figure 4. List of myxomycetes collected in the two forest sites in Mindoro Island, Philippines.

0 50 100 150 200

Ceratiomyxa fruticulosaClastoderma debaryanum

Echinostelium minutumCribraria microcarpa

Cribraria violaceaArcyria cinerea

Arcyria denudataArcyria sp.

Hemitrichia calyculataHemitrichia serpula

Metatrichia vespariaPerichaena chrysosperma

Perichaena depressaPerichaena minutum

Perichaena pedataDiachea bulbillosa

Diachea leucopodiaDiderma effusum

Diderma hemisphaericumDiderma sp.

Didymium nigripesDidymium squamulosum

Physarum albumPhysarum cinereum

Physarum compressumPhysarum crateriforme

Physarum decipiensPhysarum echinosporum

Physarum javanicumPhysarum leucophaeum

Physarum melleumPhysarum nutans

Physarum oblongaPhysarum oblatumPhysarum roseum

Physarum stellatumPhysarum superbum

Physarum sp.Colaria cf arcyrionemaComatricha tenerrimaComatricha pulchella

Comatricha sp.Lamproderma scintillans

Lycogala epidendrumStemonitis axifera

Stemonitis fuscaStemonitis spendens

Stemonitis sp.

Number of Collections

Mt. Malasimbo

Mt. Siburan



page. 17

Figure 5. Representative specimens of myxomycetes collected from the lowland forest of Mt.

Siburan, Sablayan Wateshed in Occidental Mindoro: (A) C. fruticulosa, (B) C. violacea, (C) H.

calyculata, (D) C. arcyrionema, (E) M. vesparia, (F) P. oblonga, (G) P. roseum, (H) P. javanicum,

(I) P. compressum, (J) L. epidendrum, (K) P. superbum, and (L) S. axifera.

A B C

D FE

G H I

J K L

E



page. 18

Figure 6. Representative specimens of myxomycetes collected from the lowland forest of Mt.

Malasimbo in Oriental Mindoro: (A) A. cinerea, (B) A. denudata, (C) C. tenerrima, (D) H. serpula,

(E) L. scintillans, (F) D. leucopodia, (G) P. melleum, (H) T. papillata, (I) S. fusca, (J) P. cinereum,

(K) C. microcarpa, and (L) D. bulbillosa.

A B C

D E F

G H I

J K L



page. 19

Figure 7. Scanning electron micrograph of myxomycetes collected from Mindoro: (A-B) A.

denudata, yellow arrow: calyculus (C-D) P. melleum, capillitium (C) and spores (D), (E-F) C.

violacea, red arrow: peridium, yellow arrow: peridial net (G-H) S. fusca, yellow arrow: columella

and spores (H).

A B

C D

E F

G H



page. 20

Of the 48 species recorded in this project, 45 were observed in Mt. Siburan while only 28

species were recorded in Mt. Malasimbo (Table 3). A higher number of genera was therefore noted

for the same study site, i.e. 17 genera for Mt. Siburan as opposed to 14 genera for Mt. Malasimbo.

However, when the taxonomic diversity was computed between the two forest sites, a lower SG ratio,

hence, a higher taxonomic diversity was observed for Mt. Malasimbo. Looking further at these

species, highest percentage was recorded for the Order Physarales followed by the Order

Stemonitales, and by Order Trichiales (Fig. 8). This observation is true for both study sites.

Interestingly, the highest number of species in Mt. Siburan were recorded for the woody vines

(34 species) followed by twigs (27 species), aerial leaf litter (21 species) and ground leaf litter (17

species). Higher species number was also recorded for twigs (21 species) collected in Mt. Malasimbo

followed by leaf litter (15 species). However, when the taxonomic diversity was computed, the

highest taxonomic diversity was recorded for ground leaf litter (Mt. Siburan) and aerial leaf litter (Mt.

Malasimbo). Comparing further the two sites, 19 species of myxomycetes were only recorded in Mt.

Siburan while only two species were recorded in Mt. Malasimbo (Table 4). Twenty six species were

recorded for both study sites.

Table 3. Taxonomic diversity of myxomycetes in the two forest sites in Mindoro Island.

No. Of Genera No. of Species S/G

Sites

Mt. Siburan, Sabalayan 17 45 2.65

Mt. Malasimbo, Puerto Galera 14 28 2.00

Substrate Types No. Of Genera No. of Species S/G

Mt. Siburan, Sabalayan

AL 10 21 2.10

GL 10 17 1.70

TW 14 27 1.93

WV 15 34 2.27

Mt. Malasimbo, Puerto Galera

AL 9 15 1.67

GL 8 15 1.88

TW 11 21 1.91



page. 21

A. Mt. Siburan B. Mt. Malasimbo

Figure 8. Percentage of myxomycetes collected from the two mountain sites.

Table 4. Species of myxomycetes unique at each forest site in Mindoro Island.

Mt. Siburan Mt. Malasimbo Mt. Siburan + Mt. Malasimbo

Echinostelium minutum Perichaena minutum Ceratiomyxa fruticulosa

Cribraria microcarpa Diachea bulbillosa Clastoderma debaryanum

Arcyria sp. Cribraria violacea

Hemitrichia calyculata Arcyria cinerea

Metatrichia vesparia Arcyria denudata

Diderma sp. Hemitrichia serpula

Didymium nigripes Perichaena chrysosperma

Physarum compressum Perichaena depressa

Physarum crateriforme Perichaena pedata

Physarum javanicum Diachea leucopodia

Physarum leucophaeum Diderma effusum

Physarum nutans Diderma hemisphaericum

Physarum oblonga Didymium squamulosum

Physarum roseum Physarum album

Physarum superbum Physarum cinereum

Physarum sp. Physarum decipiens

Comatricha sp. Physarum echinosporum

Lycogala epidendrum Physarum melleum

Stemonitis spendens Physarum oblatum

Physarum stellatum

Colaria cf arcyrionema

Comatricha tenerrima

Comatricha pulchella

Lamproderma scintillans

Stemonitis fusca

Stemonitis sp.

Liceales (4%)

Trichiales (20%)

Physarales (49%)

Stemonitales (20%)

Liceales (4%)

Trichiales (25%)Physarales

(43%)

Stemonitales (21%)



page. 22

To determine the species diversity, it is importance to assess the abundance of the collected

myxomycetes based on the moist chamber data. Of the 48 species recorded in this study, only one

species, Arcyria cinerea, was recorded abundant in Mt. Siburan (Table 5). Four species were noted

as common, two species as occasionally occurring, and 38 species as rare. For Mt. Malasimbo, three

species, i.e. A. cinerea, Diderma hemisphaericum, and Stemonitis sp., were recorded as abundant.

Two species were recorded as common, three species as occasionally occurring, and 20 species as

rare.

When the different substrata from Mt. Malasimbo were compared, two species were abundant

on aerial leaf litter while eight species were rare (Table 6). Four species were abundant for the ground

leaf litter while nine species were noted as rare for the same substrate. For twigs, only one species

was abundant while 11 species were noted as rare. A couple of species were also recorded as either

common or abundant in one substrata, and rare in another substrata. Several species were recorded

as rare regardless of the substrata.

For Mt. Siburan, the number of abundant species were recorded as follows: three species for

aerial leaf litter, two species for ground leaf litter, two species for woody vines, and four species for

twigs (Table 7). For the rare species, 15 were recorded for AL, 9 for GL, 19 for TW and 25 for WV.

Still, Arcyria cinerea was also recorded as abundant in all collected substrata while Physarum album

and Perichaena chrysosperma were recorded as rare in all substrata.

Computing the different species diversity indices, highest species diversity was noted for

woody vines followed by twigs, then by aerial leaf litter and finally by ground leaf litter for Mt.

Siburan (Table 8). For Mt. Malasimbo, a similar pattern was also observed with twigs being the most

diverse followed by ground and aerial leaf litter. However, almost similar evenness values were noted

for the substrates collected in Mt. Siburan. These observations simply mean that though diversity

maybe high, the species were evenly distributed on the different microhabitats.

Comparing the assemblages of myxomycetes in the two study sites, a low CC value was

recorded indicating differences in their species composition (Table 9). In fact, only 26 species of the

49 recorded species were found in both study sites (Table 4). However, when the abundance values

were included in the analysis of species composition, a higher PS value of 0.70 was recorded.



page. 23

Table 5. Abundance of myxomycetes in the two forest sites in Mindoro Island.

Order Taxon Mt. Siburan Mt. Malasimbo

Ceratiomyxales Ceratiomyxa fruticulosa R R

Echinosteliales Clastoderma debaryanum R R

Echinostelium minutum R

Liceales Cribraria microcarpa O

Cribraria violacea R R

Trichiales Arcyria cinerea A A

Arcyria denudata R R

Arcyria sp. R

Hemitrichia calyculata R

Hemitrichia serpula R R

Metatrichia vesparia R

Perichaena chrysosperma R O

Perichaena depressa R R

Perichaena minutum R

Perichaena pedata C R

Physarales Diachea bulbillosa R

Diachea leucopodia R R

Diderma effusum O O

Diderma hemisphaericum C A

Diderma sp. R

Didymium nigripes R

Didymium squamulosum R R

Physarum album R R

Physarum cinereum R R

Physarum compressum R

Physarum crateriforme R

Physarum decipiens R C

Physarum echinosporum R R

Physarum javanicum R

Physarum leucophaeum R

Physarum melleum R C

Physarum nutans R

Physarum oblonga R

Physarum oblatum R R

Physarum roseum R

Physarum stellatum R R

Physarum superbum R

Physarum sp.

R



page. 24

Stemonitales Colaria cf arcyrionema R R

Comatricha tenerrima R R

Comatricha pulchella R R

Comatricha sp. R

Lamproderma scintillans C O

Lycogala epidendrum R

Stemonitis fusca R R

Stemonitis spendens R

Stemonitis sp. C A

Table 6. Abundance of myxomycetes from the different substrata collected in Mt. Malasimbo.

Order Taxon AL GL TW

Ceratiomyxales Ceratiomyxa fruticulosa R

Echinosteliales Clastoderma debaryanum R

Liceales Cribraria violacea O

Trichiales Arcyria cinerea A A A

Arcyria denudata R

Hemitrichia serpula R

Perichaena chrysosperma O C

Perichaena depressa C

Perichaena minutum R R

Perichaena pedata R R O

Physarales Diachea bulbillosa R R

Diachea leucopodia R C

Diderma effusum O A R

Diderma hemisphaericum A A R

Didymium squamulosum O O

Physarum album R R


Physarum decipiens C

Physarum echinosporum R

Physarum melleum C A C

Physarum oblatum R

Physarum stellatum R R R

Stemonitales Colaria cf arcyrionema R

Comatricha tenerrima R C

Comatricha pulchella R R

Lamproderma scintillans C C

Stemonitis fusca R

Stemonitis sp. R C



page. 25

Table 7. Abundance of myxomycetes from the different substrata collected in Mt. Siburan, Sablayan.

Order Taxon AL GL TW WV

Ceratiomyxales Ceratiomyxa fruticulosa O R

Echinosteliales Clastoderma debaryanum R R R

Echinostelium minutum R R

Liceales Cribraria microcarpa A C

Cribraria violacea R R O

Trichiales Arcyria cinerea A A A A

Arcyria denudata A O

Arcyria sp. R

Hemitrichia calyculata R R R

Hemitrichia serpula R O R

Metatrichia vesparia R R

Perichaena chrysosperma R R R R

Perichaena depressa A R

Perichaena pedata C O R

Physarales Diachea leucopodia R O

Diderma effusum R C R

Diderma hemisphaericum A A R

Diderma sp. R

Didymium nigripes R R

Didymium squamulosum R O R

Physarum album R R R R


Physarum compressum R R

Physarum crateriforme R R

Physarum decipiens R R R

Physarum echinosporum C R

Physarum javanicum R

Physarum leucophaeum R R

Physarum melleum O R O

Physarum nutans R

Physarum oblonga R

Physarum oblatum R

Physarum roseum

Physarum stellatum O R R

Physarum superbum R R

Physarum sp. R



page. 26

Stemonitales Colaria cf arcyrionema R R

Comatricha tenerrima R O

Comatricha pulchella R R R

Comatricha sp. R

Lamproderma scintillans O C C O

Lycogala epidendrum R

Stemonitis fusca R R O

Stemonitis spendens R

Stemonitis sp. A A

Table 8. Species diversity of myxomycetes in the two forest sites in Mindoro Island

Study sites HS HG E

Mt. Siburan 1.27 6.94 0.46

Mt. Malasimbo 1.05 4.56 0.41

Substrate Types HS HG E

Mt. Siburan

AL 0.99 4.06 0.46

GL 0.98 3.68 0.52

TW 1.12 5.48 0.54

WV 1.20 6.21 0.52

Mt. Malasimbo

AL 0.83 2.81 0.38

GL 0.92 3.32 0.50

TW 1.03 3.95 0.47

Table 9. Community analysis of myxomycetes in the two forest sites in Mindoro Island

Mt. Siburan Mt. Malasimbo

Mt. Siburan 0.70 PS Value

Mt. Malasimbo 0.35 CC Value



page. 27

B. Macrofungi of Mt. Siburan and Mt. Malasimbo, Mindoro Island, Philippines

Macrofungi can either be saprophytic, parasitic or symbiotic. As saprophytes, macrofungi

grow on different types of substrates such as decomposing plant parts and leaf litter. Parasitic fungi

live within a host organism while mycorrhizal fungi form symbiotic associations with plants. Fungi

can also be ecological indicators that show significant information about the ecosystem (Eusebio,

1998). In spite of their important role in nature, the diversity of macrofungal species particularly in

the Philippines is still scantily studied. For example, Tadiosa et al. (2011) identified 38 families, 68

genera, and 107 species of macrofungi in Aurora Province, Central Luzon. Similarly, Tadiosa and

Briones (2013) recorded 75 species from 36 genera and 23 families in Batangas, Southern Luzon. De

Leon et al. (2013) likewise identified 76 species of macrofungi from Central Luzon. In this study, a

total of 34 species belonging to 21 genera and 13 families were recorded from lowland mountain

forests in Mt. Siburan in Sablayan, Occidental Mindoro and in Mt. Malasimbo in Puerto Galera,

Oriental Mindoro (Table 10). Sixteen species were exclusively reported in Mt. Siburan while 11

species were noted from Mt. Malasimbo (Table 11). Six species were identified in both study sites.

Among the collected macrofungi, six species were identified as belonging to the Division

Ascomycota (Fig. 9). These macrofungi produced ascospores within its cup-shaped ascoscarp.

Majority of the collected macrofungi belong to the Division Basidiomycota (Fig. 10). These fungal

species mainly grew on decayed woods and twigs, indicating their ability to degrade woody substrata.

To identify these macrofungi in this study, different morphological characters were observed

and recorded, i.e. description of pileus (diameter, shape, apex, surface, color, peeling, and margin),

description of lamellae (gills, attachment, arrangement) and description of stipe (color, height, width,

shape, attachment to cap, surface, annulus, and attachment to substrate and volva. However, these

characters are mainly useful to members of the agarics or the gilled mushrooms. Other morphological

characters were noted in order to identify other macrofungi. For example, for bracket fungi or

members of the family Polysporaceae, their attachment to substrata as well as their pores description

were used to characterize and identify species. It is for this reason that molecular methods were used

to confirm the identities of selected macrofungi. Here, we used the ITS sequences, a barcoding gene,

to show the phylogenetic position of the collected fungi, and thus, provide additional data to confirm

species identity.



page. 28

Table 10. List of fungal species collected in Mt. Malasimbo, Puerto Galera and Mt Siburan, Sablayan.

Family Species/Taxa Sablayan 1

(Oct 2014)

Sablayan 2

(June 2015)

Malasimbo 1

(Oct 2014)

Malasimbo 2

(June 2015)

Auriculariaceae Auricularia sp. + + + -

Agaricaceae Lycogalopsis solmsii - - + -

Ganodermataceae Ganoderma applanatum + - + -

Ganoderma lucidum + - + -

Geastraceae Geastrum mirabile + - - -

Fomitopsidaceae Fomitopsis rhodophaea - - + -

Fomitopsis sp. - + - -

Lasiosphaeriaceae Cercophora caudata - - - +

Meruliaceae Cymatoderma

dendriticum - + - -

Cymatoderma elegans - + - -

Podoscypha bolleana - + - -

Podoscypha vespillonea + - - -

Nidulariaceae Cyathus annulatus - + - -

Polyporaceae Favolus acervatus - - - +

Hexagonia tenuis + - + -

Microporus vernicipes + - + -

Microporus xanthopus - - - +

Nigroporus sp. - + - -

Perenniporia sp. - - - +

Polyporus

grammocephalus - - + -

Polyporus sp. + - - -

Polyporus tenuiculus - - + -

Pycnoporus coccineus - + - -

Pycnoporus sanguineus - - - +

Trametes sp. - + - -

Trametes versicolor - + - -

Sarcoscyphaceae Cookeina insititia - - + -

Cookeina speciosa + - - -

Cookeina tricholoma + - - -

Schizophyllaceae Schizophyllum commune + - + +

Stereaceae Xylobolus sp. - - + -

Xylariaceae Xylaria atrosphaerica - + - -

Xylaria laevis - + - -



page. 29

Table 11. Similarities in fungal composition between Mt. Malasimbo and Mt Siburan.

Mt. Siburan Mt. Siburan + Mt. Malasimbo Mt. Malasimbo

Geastrum mirabile Auricularia sp. Lycogalopsis solmsii

Fomitopsis sp. Ganoderma applanatum Fomitopsis rhodophaea

Cymatoderma dendriticum Ganoderma lucidum Cercophora caudata

Cymatoderma elegans Hexagonia tenuis Favolus acervatus

Podoscypha bolleana Microporus vernicipes Microporus xanthopus

Podoscypha vespillonea Schizophyllum commune Perenniporia sp.

Cyathus annulatus Polyporus grammocephalus

Nigroporus sp. Polyporus tenuiculus

Polyporus sp. Pycnoporus sanguineus

Pycnoporus coccineus Cookeina insititia

Trametes sp. Xylobolus sp.

Trametes versicolor

Cookeina speciosa

Cookeina tricholoma

Xylaria atrosphaerica

Xylaria laevis

Figure 9. Representative specimens of Ascomycetes collected from Mt. Siburan, Sablayan

Watershed Forest Reserve and Mt. Malasimbo, Puerto Galera in Mindoro Island: (A) C. caudate, (B)

C. tricholoma, and (C) X. laevis.

A B C



page. 30

Figure 10. Representative specimens of Basidiomycetes collected from Mt. Siburan, Sablayan

Watershed Forest Reserve and Mt. Malasimbo, Puerto Galera in Mindoro Island: (A) A. auricula, (B)

C. dendriticum, (C) Fomitopsis sp. (D) G. mirabile, (E) Perenniporia sp., (F) P. bolleana, (G) P.

tenuiculus, and (H) P. coccineus.

In this study, ITS genes of three specimens belonging to the genus Cookeina were sequenced,

aligned, and evaluated. Results of the phylogenetic analysis showed high bootstrap support. The

specimens were identified as Cookeina insititia, C. speciosa, and C. tricholoma (Fig. 11). A similar

result was also noted for specimens identified as belonging to the genera Xylaria, Podoscypha and

Cymatoderma (Fig. 12). Here, the fungal specimens were confirmed as Xylaria laevis, Podoscypha

bolleana and Cymatoderma dendriticum. One species of Xylaria could not be identified with great

certainty. Fig. 13 also showed phylogenetic tree showing closer relationship between species of

Cyathus including one specimen collected in this study. However, the lower bootstrap support could

not identify the species with high certainty. It is clear though that Polyporus gramocephalus, P.

tenuiculus, and Favolus acervatus could be identified with high certainty as supported by its 100%

bootstrap values. Indeed molecular methods coupled with morphological characterization can

accurately identify species of macrofungi.

A

B C D

H E

F

G

A



page. 31

Figure 11. Maximum likelihood tree generated for the collected specimens of the genus Cookeina.



page. 32

A. Xylaria

B. Cymatoderma

Figure 12. Maximum likelihood tree generated for specimens identified as belonging to the genera

Xylaria, Podoscypha and Cymatoderma.



page. 33

A. Genus Cyathus

Figure 14. Maximum likelihood tree for the genus Polyporus

B. Genus Polyporus

Figure 12. Maximum likelihood tree generated for specimens identified as belonging to the genera

Cyathus and Polyporus.



page. 34

V. Information Dissemination and Extension Activities

Part of the research study was presented by the proponent, Thomas Edison E. dela Cruz, at

the 8th Southeast Asia Biosphere Reserves Network (SeaBRnet) meeting and the 2nd Asia-Pacific

Biosphere Reserves Network (APBRN) Strategic Meeting on 15-19 December 2014 in Siem Reap,

Cambodia. Jointly sponsored by UNESCO, the event also included the Asia-Pacific Workshop on

Strengthening Capacity for Management of Biosphere Reserves and Protected Areas. The graduate

student, Bryna Thezza D. Leaño, also presented part of the research output under Symposium 1

(Diversity, Phylogeny and Systematics) at the Asian Mycological Congress held in Main Hall Achlya,

Goa University, Goa, India on 06 - 09 October 2015.

Figure 13. TEE dela Cruz presenting the research output at the 8th SeaBRnet and the 2nd APBRN

Meeting in Siem Reap, Cambodia.



page. 35

Figure 14. BTD Leaño presenting part of the research output on macrofungi during the Asian

Mycological Congress in Goa, India.

As part of our extension activities for the promotion of biodiversity conservation, the UST

Research Center for the Natural and Applied Sciences (RCNAS) - Fungal Biodiversity and

Systematics (FBS) group in cooperation with the Department of Biological Sciences, College of

Science, UST and the Philippine Science High School – Bicol Region Campus (PSHS-BRC),

conducted a teacher training activity on June 05-06, 2015. With the theme “Myxomycetes and

Measuring Biodiversity”, the proponent together with a visiting professor from the University of

Greifswald, Prof. Dr. Martin Schnittler, conducted a seminar-workshop on myxomycete

identification and techniques to assess and evaluate species diversity. The seminar-workshop also

included field demonstrations on collecting myxomycetes as well as a hands-on training on basic

techniques in microbiology. During this seminar-workshop, 37 high school and college teacher

participants from the Bicol Region also learned the Expedition Mundus game, a fun, educational

game that familiarize students with scientific research. The two-day seminar-workshop was also

attended by 30 high school students from six secondary schools in the region. Graduate students,

Melissa Pecundo, Nikki Heherson Dagamac, and Carlo Chris Apurillo also facilitated this activity.



page. 36

A similar one-day lecture seminar was also delivered by Prof. Dr. Martin Schnittler at the

University of Santo Tomas in Manila on June 11, 2015. In this one-day lecture-seminar, topics on

myxomycetes and measuring biodiversity were discussed to more than 70 participants from UST and

other universities including guests coming as far as Pangasinan in Northern Philippines.

Figure 15. M Schnittler delivering lectures at the University of Santo Tomas in Manila, Philippines.

The proponent organized this one-day lecture-seminar on myxomycetes and biodiversity assessment.



page. 37

Figure 16. Participants and trainors during the two-day seminar-workshop on myxomycetes and

measuring biodiversity held in Philippine Science High School – Bicol Region Campus.



page. 38

VI. Acknowledgements

This research project will not be completed without the support of the UNESCO Man and the

Biosphere programme. TEE dela Cruz thank the UNESCO MAB, the UST Research Center for the

Natural and Applied Sciences, and the graduate students, Melissa H. Pecundo and Bryna Thezza D.

Leaño. TEE dela Cruz also acknowledge the support of Dr. Martin Schnittler (University of

Greifswald, Germany), Nikki Heherson Dagamac (University of Greifswald, Germany), Dr. Young

Woon Lim (Seoul National University, Korea) Carlo Chris Apurillo, Dr. Jaycee Augusto Paguirigan,

Rio Frances Callores, Arfel Tayona and Edward dela Cruz for their assistance during the field

collection. MH Pecundo and BTD Leaño thank the Department of Science and Technology - National

Science Consortium for the graduate scholarship. The proponent also thank the local government

units of Sablayan and Puerto Galera for their assistance during the field collection.

VII. References

Adl, S.M., Simpson, A.G., Farmer, M.A., Andersen R.A., Anderson, O.R., Barta J.R., Bowser S.S.,

Brugerolle G., et al. (2005). The new higher level classification of eukaryotes with emphasis on

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