UNESCO Man and the Biosphere Programme
Young Scientists Award 2014
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 lessdocumented 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 BazalBaubo 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 24hectare 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).
Mt. Malasimbo, San Isidro, Puerto Galera
13°28’30”N, 120°54’59”E
13°28’19”N, 120°54’41”E
Sablayan Watershed, Malisbong, Sablayan
12°49’40.7”N, 120°54’57.1”E
12°49’2.3”N, 120°53’48.5”E
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% (PAGASA, DOST).
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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
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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)
Equation 1:
R
HS = −
∑ i (pilnpi)
i=1
where R = species count
ii.
pi = proportion of R represented by the ith species
Gleason Index of Species Richness (HG)
Equation 2:
HG =
Np-1
lnNi
lnNi
lnNi
where Np= total number of species
Ni= total number of individuals in the ith species
iii.
Pielou’s Index of Species Evenness (E)
Equation 3:
Hs
E=
Hmax
where HS= Shannon Index of Diversity
Hmax= the maximum value of HS
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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
Ceratiomyxa fruticulosa var
arbuscola
Didymium nigripes
Didymium squamulosum
Cribraria cancellata
Cribraria microcarpa
Perichaena chrysosperma
Didymium nigripes
Arcyria denudate
Stemonitis fusca
Physarum stellatum
decaying bamboo
Number of Records
dried fruit still attached to the plant
ground litter
twigs
decayed wood
twigs
ground litter
decaying bamboo
decayed wood
decayed wood
1
1
2
1
1
1
1
2
1
1
decayed wood
decayed wood
bark
ground litter
ground litter
ground litter
decayed wood
decayed wood
ground litter
twigs
decayed wood
1
1
1
2
1
2
1
2
1
1
1
Mt. Malasimbo
Stemonitis sp.
Hemitrichia serpula
Physarum leucophaeum
Diachea leucopodia
Didymium iridis
Craeterium leucophaeum
Arcyria denudata
Arcyria cinerea
Didymium squamulosum
Hemitrichia calyculata
Physarum viride
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Table 2. Productivity of moist chambers.
STUDY SITES
Mt. Siburan
Mt. Malasimbo
SUBSTRATE TYPES
Mt. Siburan
AL
GL
TW
WV
Mt. Malasimbo
AL
GL
TW
PLASMODIUM
17%
32%
FRUITING BODY
49%
48%
MYXOMYCETES
65%
80%
PLASMODIUM
24%
14%
19%
8%
PLASMODIUM
32%
3%
27%
FRUITING BODY
55%
36%
49%
66%
FRUITING BODY
55%
30%
59%
MYXOMYCETES
79%
50%
68%
73%
MYXOMYCETES
87%
66%
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).
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Stemonitis sp.
Stemonitis spendens
Stemonitis fusca
Stemonitis axifera
Lycogala epidendrum
Lamproderma scintillans
Comatricha sp.
Comatricha pulchella
Comatricha tenerrima
Colaria cf arcyrionema
Physarum sp.
Physarum superbum
Physarum stellatum
Physarum roseum
Physarum oblatum
Physarum oblonga
Physarum nutans
Physarum melleum
Physarum leucophaeum
Physarum javanicum
Physarum echinosporum
Physarum decipiens
Physarum crateriforme
Physarum compressum
Physarum cinereum
Physarum album
Didymium squamulosum
Didymium nigripes
Diderma sp.
Diderma hemisphaericum
Diderma effusum
Diachea leucopodia
Diachea bulbillosa
Perichaena pedata
Perichaena minutum
Perichaena depressa
Perichaena chrysosperma
Metatrichia vesparia
Hemitrichia serpula
Hemitrichia calyculata
Arcyria sp.
Arcyria denudata
Arcyria cinerea
Cribraria violacea
Cribraria microcarpa
Echinostelium minutum
Clastoderma debaryanum
Ceratiomyxa fruticulosa
Mt. Malasimbo
Mt. Siburan
0
50
100
150
200
Number of Collections
Figure 4. List of myxomycetes collected in the two forest sites in Mindoro Island, Philippines.
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A
B
C
D
EE
F
G
H
I
J
K
L
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.
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A
B
C
D
E
F
G
H
I
J
K
L
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.
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A
B
C
D
E
F
G
H
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).
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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
Mt. Siburan, Sabalayan
Mt. Malasimbo, Puerto Galera
17
14
45
28
2.65
2.00
Substrate Types
Mt. Siburan, Sabalayan
AL
GL
TW
WV
Mt. Malasimbo, Puerto Galera
AL
GL
TW
No. Of Genera
No. of Species
S/G
10
10
14
15
21
17
27
34
2.10
1.70
1.93
2.27
9
8
11
15
15
21
1.67
1.88
1.91
Sites
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Liceales
(4%)
Liceales
(4%)
Trichiales
(20%)
Trichiales
(25%)
Physarales
(43%)
Physarales
(49%)
Stemonitales
(20%)
A. Mt. Siburan
Stemonitales
(21%)
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
Echinostelium minutum
Cribraria microcarpa
Arcyria sp.
Hemitrichia calyculata
Metatrichia vesparia
Diderma sp.
Didymium nigripes
Physarum compressum
Physarum crateriforme
Physarum javanicum
Physarum leucophaeum
Physarum nutans
Physarum oblonga
Physarum roseum
Physarum superbum
Physarum sp.
Comatricha sp.
Lycogala epidendrum
Stemonitis spendens
Mt. Malasimbo
Perichaena minutum
Diachea bulbillosa
Mt. Siburan + Mt. Malasimbo
Ceratiomyxa fruticulosa
Clastoderma debaryanum
Cribraria violacea
Arcyria cinerea
Arcyria denudata
Hemitrichia serpula
Perichaena chrysosperma
Perichaena depressa
Perichaena pedata
Diachea leucopodia
Diderma effusum
Diderma hemisphaericum
Didymium squamulosum
Physarum album
Physarum cinereum
Physarum decipiens
Physarum echinosporum
Physarum melleum
Physarum oblatum
Physarum stellatum
Colaria cf arcyrionema
Comatricha tenerrima
Comatricha pulchella
Lamproderma scintillans
Stemonitis fusca
Stemonitis sp.
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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.
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Table 5. Abundance of myxomycetes in the two forest sites in Mindoro Island.
Order
Ceratiomyxales
Echinosteliales
Liceales
Trichiales
Physarales
Mt. Siburan
Mt. Malasimbo
Ceratiomyxa fruticulosa
Clastoderma debaryanum
Echinostelium minutum
Cribraria microcarpa
Cribraria violacea
Arcyria cinerea
Arcyria denudata
Arcyria sp.
Hemitrichia calyculata
Hemitrichia serpula
Metatrichia vesparia
Perichaena chrysosperma
Perichaena depressa
Perichaena minutum
Perichaena pedata
Diachea bulbillosa
Diachea leucopodia
Diderma effusum
Diderma hemisphaericum
Diderma sp.
Didymium nigripes
Didymium squamulosum
Physarum album
Physarum cinereum
Physarum compressum
Physarum crateriforme
Physarum decipiens
Physarum echinosporum
Physarum javanicum
Physarum leucophaeum
Physarum melleum
Physarum nutans
Physarum oblonga
Physarum oblatum
Physarum roseum
Physarum stellatum
Physarum superbum
Taxon
R
R
R
O
R
A
R
R
R
R
R
R
R
R
R
Physarum sp.
R
C
R
O
C
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
R
A
R
R
O
R
R
R
R
R
O
A
R
R
R
C
R
C
R
R
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Stemonitales
Colaria cf arcyrionema
Comatricha tenerrima
Comatricha pulchella
Comatricha sp.
Lamproderma scintillans
Lycogala epidendrum
Stemonitis fusca
Stemonitis spendens
Stemonitis sp.
R
R
R
R
C
R
R
R
C
R
R
R
O
R
A
Table 6. Abundance of myxomycetes from the different substrata collected in Mt. Malasimbo.
Order
Taxon
Ceratiomyxales
Ceratiomyxa fruticulosa
Echinosteliales
Clastoderma debaryanum
Liceales
Cribraria violacea
Trichiales
Arcyria cinerea
AL
GL
R
R
O
A
A
Arcyria denudata
R
O
C
Perichaena depressa
Physarales
C
Perichaena minutum
R
R
Perichaena pedata
R
R
Diachea bulbillosa
R
R
Diachea leucopodia
R
C
Diderma effusum
O
A
R
Diderma hemisphaericum
A
A
R
Didymium squamulosum
O
O
O
Physarum album
R
R
Physarum cinereum
R
R
Physarum decipiens
C
Physarum echinosporum
Physarum melleum
R
C
A
Physarum oblatum
Physarum stellatum
Stemonitales
A
R
Hemitrichia serpula
Perichaena chrysosperma
TW
R
R
R
Colaria cf arcyrionema
R
R
Comatricha tenerrima
R
Comatricha pulchella
R
Lamproderma scintillans
C
C
R
C
Stemonitis fusca
Stemonitis sp.
C
R
R
C
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Table 7. Abundance of myxomycetes from the different substrata collected in Mt. Siburan, Sablayan.
Order
Taxon
TW
WV
O
R
R
R
Echinostelium minutum
R
R
Cribraria microcarpa
A
C
Ceratiomyxales
Ceratiomyxa fruticulosa
Echinosteliales
Clastoderma debaryanum
Liceales
Trichiales
AL
GL
R
Cribraria violacea
R
Arcyria cinerea
A
A
Arcyria denudata
R
O
A
A
A
O
Arcyria sp.
R
Hemitrichia calyculata
R
R
R
Hemitrichia serpula
R
O
R
Metatrichia vesparia
R
R
Perichaena chrysosperma
R
Perichaena depressa
A
Perichaena pedata
Physarales
R
R
C
O
R
Diachea leucopodia
R
O
Diderma effusum
R
C
Diderma hemisphaericum
A
A
Diderma sp.
R
R
R
R
Didymium nigripes
R
Didymium squamulosum
R
O
Physarum album
R
R
Physarum cinereum
R
R
R
R
Physarum compressum
R
Physarum decipiens
R
Physarum echinosporum
R
R
R
Physarum crateriforme
R
R
R
C
Physarum javanicum
R
R
R
Physarum leucophaeum
R
Physarum melleum
O
Physarum nutans
R
R
R
O
Physarum oblonga
Physarum oblatum
R
R
R
Physarum roseum
Physarum stellatum
Physarum superbum
Physarum sp.
O
R
R
R
R
R
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Stemonitales
Colaria cf arcyrionema
R
R
Comatricha tenerrima
R
O
R
R
C
O
Comatricha pulchella
R
Comatricha sp.
Lamproderma scintillans
R
O
C
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
Substrate Types
HS
1.27
1.05
HS
HG
6.94
4.56
HG
E
0.46
0.41
E
AL
GL
TW
WV
0.99
0.98
1.12
1.20
4.06
3.68
5.48
6.21
0.46
0.52
0.54
0.52
AL
GL
TW
0.83
0.92
1.03
2.81
3.32
3.95
0.38
0.50
0.47
Mt. Siburan
Mt. Malasimbo
Mt. Siburan
Mt. Malasimbo
Table 9. Community analysis of myxomycetes in the two forest sites in Mindoro Island
Mt. Siburan
Mt. Siburan
Mt. Malasimbo
Mt. Malasimbo
0.70
0.35
PS Value
CC Value
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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.
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Table 10. List of fungal species collected in Mt. Malasimbo, Puerto Galera and Mt Siburan, Sablayan.
Family
Species/Taxa
Auriculariaceae
Agaricaceae
Ganodermataceae
Auricularia sp.
Lycogalopsis solmsii
Ganoderma applanatum
Ganoderma lucidum
Geastrum mirabile
Fomitopsis rhodophaea
Fomitopsis sp.
Cercophora caudata
Cymatoderma
dendriticum
Cymatoderma elegans
Podoscypha bolleana
Podoscypha vespillonea
Cyathus annulatus
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
Cookeina insititia
Cookeina speciosa
Cookeina tricholoma
Schizophyllum commune
Xylobolus sp.
Xylaria atrosphaerica
Xylaria laevis
Geastraceae
Fomitopsidaceae
Lasiosphaeriaceae
Meruliaceae
Nidulariaceae
Polyporaceae
Sarcoscyphaceae
Schizophyllaceae
Stereaceae
Xylariaceae
Sablayan 1
(Oct 2014)
Sablayan 2
(June 2015)
Malasimbo 1
(Oct 2014)
Malasimbo 2
(June 2015)
+
+
-
-
+
+
+
+
-
+
+
+
-
+
-
+
-
-
+
-
+
-
-
-
+
+
+
-
-
+
-
-
+
+
+
-
+
+
+
-
-
+
-
-
-
-
+
-
+
-
-
-
-
+
+
+
-
+
-
+
+
+
-
-
+
+
+
-
-
+
+
-
+
+
-
+
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Table 11. Similarities in fungal composition between Mt. Malasimbo and Mt Siburan.
Mt. Siburan
Geastrum mirabile
Fomitopsis sp.
Cymatoderma dendriticum
Cymatoderma elegans
Podoscypha bolleana
Podoscypha vespillonea
Cyathus annulatus
Nigroporus sp.
Polyporus sp.
Pycnoporus coccineus
Trametes sp.
Trametes versicolor
Cookeina speciosa
Cookeina tricholoma
Xylaria atrosphaerica
Xylaria laevis
A
Mt. Siburan + Mt. Malasimbo
Auricularia sp.
Ganoderma applanatum
Ganoderma lucidum
Hexagonia tenuis
Microporus vernicipes
Schizophyllum commune
B
Mt. Malasimbo
Lycogalopsis solmsii
Fomitopsis rhodophaea
Cercophora caudata
Favolus acervatus
Microporus xanthopus
Perenniporia sp.
Polyporus grammocephalus
Polyporus tenuiculus
Pycnoporus sanguineus
Cookeina insititia
Xylobolus sp.
C
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.
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B
A
F
E
C
G
D
H
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.
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Figure 11. Maximum likelihood tree generated for the collected specimens of the genus Cookeina.
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A. Xylaria
B. Cymatoderma
Figure 12. Maximum likelihood tree generated for specimens identified as belonging to the genera
Xylaria, Podoscypha and Cymatoderma.
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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.
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V.
Information Dissemination and Extension Activities
Part of the research study was presented by the proponent, Thomas Edison E. dela Cruz, at
the 8 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.
th
Figure 13. TEE dela Cruz presenting the research output at the 8th SeaBRnet and the 2nd APBRN
Meeting in Siem Reap, Cambodia.
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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.
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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.
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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.
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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.
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