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"ABSTRACTMarasmiellus is a tropical and subtropical genus which consists of more than 400 species. The genus plays important roles in ecosystem as a decomposer, and not many information on its economic value. Several monographs of the genus have been published based on morphological data from several tropical forests (neotropics, Africa, Srilanka), and no report on the Indonesian Marasmiellus. At present, taxonomic problem occurs in the genus related to the position of Marasmiellus juniperinus as a type genus in the phylogenetic tree based on Internal Transcribed Spacer (ITS) region. Marasmiellus juniperinus nested into different clade from other Marasmiellus species, and it was clustered in Gymnopus clade. Since Marasmiellus has never been reported from Indonesia and the taxonomic problem occurs at the genus, thus the objectives of this study are 1) To describe the species of Marasmiellus found in Java and Bali; 2) To provide an identification key to sections and species; 3) To analyse the phylogenetic relationship within the genus Marasmiellus in Java and Bali based on morphological and molecular characters performed by Maximum Parsimony method; 4) To clarify the relationship of the genus Marasmiellus and M. juniperinus to its closely related genera with inclusion of more Marasmiellus spp. based on ITS region of rDNA sequence data; and 5) To describe novel species of Marasmiellus based on morphological and molecular data. Java and Bali were chosen as research sites in this study due to several reasons. First, historically most of Indonesian reported agarics were collected from Java, particularly from Mount Gede-Pangrango, Cibodas and Bogor Botanical Gardens. Second, forest degradation is going rapidly in Java and Bali. As a result Indonesia is loosing habitats of Marasmiellus. This study was divided into three topics according to the purposes of this study. The first topic entitled Species of Marasmiellus in Java and Bali. The study was carried out in Herbarium Bogoriense, The Botany Division, Research Center for Biology, The Indonesian Institute of Sciences (LIPI). Herbarium materials are kept in Herbarium Bogoriense (BO) and Harry D. Thiers Herbarium, San Francisco State University (SFSU), SF, CA, USA.Materials used in this research were collected in 1998-2010 from 10 different locations in Java and Bali which consisted of 22 sites. A representative material of Marasmiellus juniperinus, as a type genus, was borrowed from the Herbarium of the University of Tennessee, Knoxville, Tennessee, USA (TENN). The identification results revealed that there were 35 known species; one species as a new combination; 17 undescribed species. Those 35 species consist of 25 species of Marasmiellus found in Java, 7 species found in Bali, and 3 species found both in Java and Bali. All described species were treated based on infrageneric classification of Singer (1973) which divided the genus into 10 sections based on morphological characters.Based on Singer?s infrageneric classification Marasmiellus in Java and Bali belonged to 5 sections, i.e sect. Marasmiellus, sect. Rameales, sect. Dealbati, sect. Candidi, and sect. Stenophylloides. The second topic entitled Phylogenetic study of the genus Marasmiellus based on morphological and molecular analysis. Thirty five morphological characters of 37 taxa were scored manually for phylogenetic analysis based on morphological characters. Those taxa consisted of 35 species of Marasmiellus in Java and Bali found (topic 1), a type species of Marasmiellus (M. juniperinus), and Crinipellis brunneipurpurea as an outgroup. Matrix data was analyzed by Phylogenetic Analysis Using Parsimony (PAUP) program. The topology of phylogenetic tree based on morphological data was compared to topology of phylogenetic tree based on molecular data. The phylogenetic tree based on morphological and molecular characters showed that Marasmiellus spp. were divided into four major clades. Each clade consisted of several Singer?s sections. This result indicated that the sections within Singer?s infrageneric classification were polyphyletic. Morphological and molecular phylogenies resulted in this study did not support traditional classification, i.e. Singer?s classification of Marasmiellus. The phylogenetic relationship of Marasmiellus taxa, Gymnopus and its closely related genera were analyzed using molecular data of ITS rDNA. Forty one sequences based on ITS region representing of 36 species of Marasmiellus were used for phylogenetic analysis. Nine sequences of Marasmiellus were generated from this study, and 32 sequences were obtained from the international DNA database (GenBank). The phylogenetic tree showed that Marasmiellus taxa were divided into 4 clades. The taxa of Marasmiellus and Gymnopus were clustered together in three of four clades. The type species, Marasmiellus juniperinus was located within the same clade as Gymnopus fusipes.This result indicated that the genus Marasmiellus was polyphyletic. The third topic entitled Description of a novel species, Marasmiellus javanicus, based on morphological and molecular analysis. Marasmiellus javanicus is one of the 17 new species in this study, and it was described by using the integrated morphological and molecular data. Marasmiellus javanicus was described as a new species based on 5 specimens found in Bogor Botanical Garden, West Java. Morphologically it was similar to M. hondurensis (Murrill) Singer which was collected from Belice in 1906 by Peck (Singer 1973). The characters similarities of M. javanicus and M. hondurensis were off-white, same size of width, convex with flattened disc of pileus; subdistant lamellae; tomentose and insititious stipe; cutis with diverticulate of pileipellis, and presence of stipe vesture. Marasmiellus hondurensis was different from M. javanicus in having longer stipe (6?22 mm diam), fusoid basidiospores, Rotalis-type of cheilocystidia, and M. hondurensis was found on dicotyledon plant. At present, there is no sequence based on ITS rDNA of M. hondurensis. Thus, genetically M. hondurensis was unable to be compared to M. javanicus. To know the closest species of M. javanicus, phylogenetic tree of Marasmiellus was constructed. Phylogenetic tree showed that the most closely related species to M. javanicus was M. mesosporus. Marasmiellus javanicus and M. mesosporus have ITS rDNA 94% sequence similarity. They differ in 27 nucleotides which were 7 insertion and deletion, and 24 of substitution (A↔G=11, C↔T=13). This study showed that integrated morphological and molecular data are required to identify the Marasmiellus."

"Polisakarida intrasel adalah penyusun dinding sel hifa dan karpus cendawan yang menunjukkan aktivitas farmakologi yang cukup luas. Polisakarida cendawan umumnya diproduksi dengan mengekstrak dari karpus. Namun dibutuhkan waktu beberapa bulan untuk menumbuhkan hifa dari inokulum hingga karpus cendawan. Oleh karena itu diupayakan alternatif produksi polisakarida cendawan langsung dari miselia, melalui fermentasi cair. Tujuan dari penelitian ini adalah untuk mengetahui kondisi optimal produksi polisakarida intrasel dari miselia cendawan tiram putih melalui fermentasi cair, dan mengetahui aktivitas imunomodulatornya. Faktor-faktor yang dioptimasi pada penelitian ini adalah: suhu, pH, agitasi dan waktu inkubasi, dengan desain central composite design (CCD) dari metode respon permukaan (RSM). Hasil penelitian menunjukkan kondisi optimal fermentasi cair P. ostreatus BPPTCC 6017 untuk menghasilkan biomassa dan polisakarida yang terbaik adalah pada suhu 27,89°C, pH awal medium of 5,49, agitasi 124,08 rpm, dan waktu fermentasi 11,44 hari yang menghasilkan biomassa kering 32,00±1,25 g/l, polisakarida intrasel larut air 0,29±0,01 g/l (rendemen 0,91%), dan polisakarida intrasel larut alkali 0,60±0,02 g/l (rendemen 1,88%), yang mendekati kondisi teoritik. Aktivitas imunomodulator polisakarida intrasel larut air dan larut alkali miselium P. ostreatus BPPTCC 6017 diujikan pada sistem imun alamiah berdasar aktivitas dan kapasitas fagositosis makrofag peritoneum mencit secara in vitro, bersihan karbon, dan sistem komplemen jalur klasik. Efek imunomodulator juga diujikan pada sistem imun adaptif sel dan humoral berdasar peningkatan jumlah limfosit total dan limfosit T, serta kadar imunoglobulin G. Hasil penelitian membuktikan bahwa polisakarida intrasel larut air dan larut alkali miselium tiram putih mempunyai aktivitas imunomodulator terhadap sistem imun alamiah dan adaptif masing-masing adalah 2 mg/20 g BB mencit dan 4 mg/20 g BB mencit atau setara 100 mg/kg BB dan 200 mg/kg BB.

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Intracellular polysaccharide is a component of fungal cell-wall that has a wide pharmacological activity. Mushroom polysaccharide commonly produced by extracting the fruit body. However, it takes several months starting by growing hyphae until forming fruit body, therefore an alternative for producing it directly from mycelia instead of fruit body through submerged fermentation. This study was aimed to determine the optimal submerged fermentation conditions for producing intracellular polysaccharide of oyster mushroom. Four variables were tested in this study: initial pH medium, agitation speed, incubation time and temperature, using a central composite design (CCD), one of the design in response surface methodology (RSM). The optimal conditions of submerged fermentation obtained as follows: 27.89°C, initial pH medium of 5.49, agitation speed of 124.08 rpm, and fermentation time of 11.44 days. Verification of optimal conditions produced 32.00±1.25 g/l dried mycelia, 0.29±0.01 g/l water-soluble POP (yield 0.91%), and 0.60±0.02 g/l alkali-soluble POP (yield 1.88%), which were close to the theoretical conditions. Immunomodulatory activity of intracellular polysaccharide then assessed on the innate immune system, based on activity and capacity phagocytic of murine peritoneum macrophages in vitro, carbon clearance, and on the classical complement pathway. Immunomodulatory assay also conducted on the cellular and humoral adaptive immune system, based on the increasing number of total lymphocyte, T lymphocyte, and Immunoglobulin G. Result of a series of immunomodulatory activity assay showed that the intracellular polysaccharides of P. ostreatus BPPTCC 6017 have immunomodulatory activity on the innate and adaptive immune system at a dose of 2 mg/20 g BW mice and 4 mg/20 g BW mice respectively, equivalent to 100 mg/kg BW and 200 mg/kg BW."