The total mushroom production in Korea in 2015 comprised about 200,000 ton with a value of about 700 billion won (one trillion won if mushroom factory products are included, 1 $=1,128 won). This is almost equal to the amounts generated by flowers and medicinal crops. Even though the cultivation area and the number of farms have been decreased since 2006, the scale of the farms is increasing. Also mushroom productivity and consumption (4.9 ㎏ / person in 2015) are steadily increasing. Major cultivated species are Flammulina velutipes, Pleurotus ostreatus, Pleurotus eryngii, Lentinula edodes and Agaricus bisporus, which cover 90% of the total production. Shelves, logs, plastic bags or plastic bottles are mostly used for cultivation depending on the kind of mushroom. Mushroom export estimated about 43 million $ in 2016. Major exported mushrooms are Flammulina velutipes, Pleurotus eryngii, Lentinula edodes, Pleurotus ostreatus, Tricholoma matsutake and Agaricus bisporus that are destined for the U.S., the Netherlands, Australia, Canada, Hong Kong, Japan and several other countries. Mushroom research in the RDA started in 1967. In the beginning, development of cultivation methods using rice straw was studied together with distribution of improved strains of the button mushroom. The costs of button mushroom production strongly increased due the improving national economy, shortage of workers, increase of wages and the oil crisis in the nation. This caused a weak competitive position for button mushroom export and resulted in reduction of the button mushroom cultivation area. After development of a cotton waste cultivation method for the oyster mushroom (Pleurotus ostreastus), this mushroom was popularized and mainly studied. Development of automated plastic bottle cultivation systems in the 1990s changed shelf cultivation to mass production. The bottle system was applied to the oyster mushroom but also to F. velutipes and P. eryngii. Mushroom research in the RDA nowadays focuses on analysis of mushroom genetics in combination with development of new mushroom varieties, mushroom physiology and cultivation. Further studied are environmental factors for cultivation, disease control, development and utilization of mushroom substrate resources, post-harvest management and improvement of marketable traits. Finally, the RDA manages the collection, classification, identification and preservation of mushroom resources.

Molecular genetic approaches have advantages in investigating various physiological aspects of mushrooms. We have been engaged in developing genetic transformation system mainly in wood rot mushrooms. In this presentation, our recent trials to accelerate mushroom science beyond the post-genomic era will be presented. They include development of research tools both in forward and reverse genetics; a transfection system and its utilization in gene expression signal analysis in the selective white rot fungus Ceriporiopsis subvermispora, multiple gene targeting system in Pleurotus ostreatus and Coprinopsis cinerea, which was used to do knock-out or knock-in of reporter constructs on the chromosome of these fungi, and a “powered” forward genetics combined with genome information was used to identify new gene responsible for lignin-degradation in P. ostreatus. They may contribute to elucidate uncovered molecular mechanisms in unique processes in the mushrooms and to make them as a new industrial tool in the next era. Furthermore, our recent results in on-going genome editing experiments will be also reported.

Ganoderma lucidum is one of the most popular medicinal mushrooms of Asian region having promising immune enhancing properties. This single species of mushroom has over 400 bioactive compounds and possess properties relating to battling tumours, viruses, bacteria, inflammation, platelet-aggregation, immune system modulation among many others. There are documented reports of G. lucidum fighting the activity against HIV/AIDS and diabetes. Owing to such a wide range of medicinal applications, the global consumption of G. lucidum is high, which has resulted in a plethora of commercially available patented products. These products have G. lucidum as an active constituent and are usually marketed as food supplements but are also available in the form of creams, hair tonics and medicinal soaps. The growing demand and the high cost of G. lucidum in the local market have enforced to think about a range of novel methods for the cultivation of this mushroom. We explored different novel strategies in this direction. In the first approach, the widely available agricultural residues of oil palm biomass (empty fruit bunch fibres, EFB) from Malaysia were used to cultivate this mushroom with cheap cost. The required conditions for the growth of this mushroom were studied by employing a pilot-scale cultivation in a mushroom farm. From the results it was scrutinized that oil palm waste fibres are the potential source of cultivating G. lucidum either alone or in combination of substrates. In the second strategy, the effect of UV-C radiation on the physical growth of G. lucidum mycelium has shown positive results. Results of 10 and 15 min of UV-C exposure treatments per 24 h showed a 30% increase in the diameter of mycelium over control in Potato Dextrose Agar medium petri-plates. Using EFB as substrates not only supports the G. lucidum cultivation but also offers a viable alternative for the management of solid waste in oil palm plantations of Malaysia. These results stipulate a better and stronger mushroom in terms of its bioactive compounds content and better growth parameters. Moreover, it also benefits the local mushroom farmers in Malaysia for the easy availability of the substrate materials and with better economy. The techniques involved can be propagated to the local Ganoderma industry for improvised cultivation for better output. A high output would also mean profitable business resulting in revolutionizing the economics of Ganoderma markets all over the world.

Lentinula edodes shows atypical mating behaviors deviated from tetrapolar mating in outcrossing. To better understand the mating behavior, we investigated variations in the mating type genes, residing in A and B mating loci, of 129 dikaryotic strains from East Asia. Through sequence analysis of A locus, we found that hypervariable region spanning N-term of HD2-intergenic region-N-term of HD1 could represent A mating type of L. edodes. Mating and hypervariable region analyses revealed 50 unique A mating types: 27 from 98 cultivated strains, 33 from 31 wild strains, and 10 commonly found. It was also revealed that only a few A mating type alleles such as A1, A4, A5, and A7 were prevalent in cultivated strains. The A mating type in wild strains were highly diverse: 23 unique A alleles were discovered in small mountainous area in Korean peninsula. The B locus was assessed by allelic variations in pheromone (PHB) and pheromone receptor (RCB) pairs. Sequence analyses revealed 5 alleles of RCB1 with 9 associated PHBs in B sublocus and 3 alleles of RCB2 with 5 associated PHBs in B sublocus. Each RCB was primarily associated with two PHBs. This allowed us to propose 15 B mating types via combinations of five B and three B subloci. Thus, the total number of mating types in L. edodes became 750 through a combination of 50 A and 15 B. This number will further increase because of rapid diversification of A mating type and possible emergence of variants of RCBs and PHB-RCB combinations.

Collybia reinakeana (P.) Henn used to be a wild endemic edible mushroom not until it was discovered in the mountainous area of Puncan, Carranglan, Nueva Ecija, Philippines after the great earthquake that jolted Central Luzon in 1990. It is characterized to having big fruiting bodies (about 1 foot tall) that grow in cluster. Its mycelia were rescued from the wild and the first domestication studies were conducted which lead to the generation of its production technology. It is a nutraceutical mushroom that exhibits antibacterial and anti-hypertensive properties aside from its appreciable content of both standard as well as non- standard amino acids. It also exhibits mycoparasitic habit against species of Aspergillus, Cladosporium and Fusarium. Recently, emergence of this mushroom in the different areas in the Philippines have been reported which prompted our research team to conduct geo – tracking in order to rescue its secondary mycelia in our desire to conserve this endemic species of mushroom. With our conservation efforts, we have tracked the areas where it was found growing and subsequently isolated the secondary mycelia. At present, our research team were able to collect 9 different strains from the different areas namely Bataan (1), Science City of Munoz (2), Urdaneta City (1), San Manuel, Pangasinan (1), Rosales, Pangasinan (1), Bicos, Rizal, Nueva Ecija (1), Dipaculao, Aurora (1) and Bamban, Tarlac(1).

In Volvariella volvacea, a species of edible mushroom, cryogenic autolysis is a typical part of abnormal metabolism. Previous functional annotation cluster analyses of cold-induced gene expression profiles have shown that the ubiquitin-conjugating enzyme E2 (UBE2) and the cyclin-like F-box domain (FBDC) form the functional clusters. Among them, a specific FBDC gene has been found to be in response to cold stress and correlated with the cryogenic autolysis of V. volvacea. In this study, analysis of gene expression profiling showed that only one type of UBE2 in V. volvacea (UBEV2) was significantly up-regulated. Further quantitative real-time PCR analysis confirmed that the expression of UBEV2 was significantly up-regulated (P < 0.05) after cold-treatment lasting 4, 6, and 8 h. Western-bolt analyses confirmed that the protein expression of UBEV2 was in response to cold stress. This provided evidence that UBEV2 was a cold stress-responsive gene and closely correlated with cryogenic autolysis. The specific distribution of UBEV2 in recently diverged herb decay fungi indicated that UBEV2 was not evolutionarily correlated with early diverging fungi. Phylogenetic analysis indicated that UBEV2 was generated by horizontal gene transfer (HGT) from the ancestry of Selaginella moellendorffii UBE2. Further relative time estimation and detection of natural selection showed that there has been recent positive selection after HGT in UBEV2. Molecular modeling and logo analysis showed that the cysteine-cysteine motif is the characteristic of the UBEV2 family. These observations indicate that UBEV2 and FBDV1 are the newly discovered cold stress-responsive genes correlated with the cryogenic autolysis of V. volvacea.

This study was carried out to determine the basic mycelial culture conditions for Poria cocos growth. According to colony diameter and mycelial density, suitable media for mycelial growth were Malt yeast extract, Potato dextrose agar, Yeast extract agar, and Yeast malt agar. The optimum temperature for mycelial growth was between 25 and 35oC, and the optimum pH value was between 4 and 7. Carbon and nitrogen sources were fructose and yeast extract. The optimum C/N ratio was about 10 to 1 with 2% glucose. Other minor components for optimal growth were thiamine-HCl and nicotinamide as vitamins, acetic and lactic acid as organic acids, and MgSO4·7H2O and FeSO4·7H2O as mineral salts. Wolfiporia cocos is a well-known traditional medicine in China, Japan, Korea, and other Asian countries owing to its numerous therapeutic properties. With the aim to determine the morphology and genetic characteristics of W. cocosten strains of W. cocos were cultivated in vitro, and subsequently, rapid amplification of polymorphic DNA was performed. To the best of our knowledge, this is the first study to examine the morphology of fruit bodies of W. cocos in Korea. W. cocos were cultured on PDA agar at different temperatures (12, 16, 20, 24, and 28oC) under 12-hour light (600 Lux) / 12-hour dark photoperiod condition for 1 month. Appearance of fruit body was the highest at 28°C condition in all the strains investigated. Honeycomb-like structure on sclerotia was observed in Andong 01, Andong 02, Andong 03, KFRI 1104, KFRI 1105, KFRI 1106, KFRI 1107, KFRI 1108, and ASI 13007 strains. The KFRI 1103 strain formed cosmos petal-like structure on sclerotia. The average size of basidiospores was recorded as 7.55 μm in height and 3.35 μ in width. This study was carried out to discriminate the geographical origin from Korea and Chinese Wolfiporia extensa. By proximate composition analysis, both were identified as similar, showed oxygen was 45.32-48.07%, carbon was 38.09-40.12%, hydrogen was 6.05~6.78% and nitrogen was 0.16-0.23%. Antioxidant activity was examined by DPPH free radical scavenging activity. No significant differences were found as well for the antioxidant activity between Korean and Chinese product. However, the contents of inorganic components ED-XRF (X-Ray Fluorescence Spectrometer) were significant different in Chinese and domestic W. extensa. The Potassium and iron in Chinese and domestic W. extensa were 47.60±8.78% and 14.5±3.86% as well as 33.14±17.27% and 9.13±4.83%, respectively. From the above resuls, the analysis of inorganic components by ED-XRF may be used for discrimination of the geographical origin of W. extensa.

Collybia reinakeana (P.) Henn used to be a wild endemic edible mushroom not until it was discovered in the mountainous area of Puncan, Carranglan, Nueva Ecija, Philippines after the great earthquake that jolted Central Luzon in 1990. It is characterized to having big fruiting bodies (about 1 foot tall) that grow in cluster. Its mycelia were rescued from the wild and the first domestication studies were conducted which lead to the generation of its production technology. It is a nutraceutical mushroom that exhibits antibacterial and anti-hypertensive properties aside from its appreciable content of both standard as well as non- standard amino acids. It also exhibits mycoparasitic habit against species of Aspergillus, Cladosporium and Fusarium. Recently, emergence of this mushroom in the different areas in the Philippines have been reported which prompted our research team to conduct geo - tracking in order to rescue its secondary mycelia in our desire to conserve this endemic species of mushroom. With our conservation efforts, we have tracked the areas where it was found growing and subsequently isolated the secondary mycelia. At present, our research team were able to collect 9 different strains from the different areas namely Bataan (1), Science City of Munoz (2), Urdaneta City (1), San Manuel, Pangasinan (1), Rosales, Pangasinan (1), Bicos, Rizal, Nueva Ecija (1), Dipaculao, Aurora (1) and Bamban, Tarlac(1).

Cordyceps species are diverse and grow worldwide from larval to adult stages of a range of insects. They are traditionally regarded as medicinal mushrooms in Asia for their immune-enhancing properties. Different species of Cordyceps are being cultivated in large scale to fulfill the demand of public consumption as supplementary health food or immune booster. Cultivation of Cordyceps fruiting bodies is a complex process and a controlled environment of temperature, light, humidity etc. along with appropriate medium is a prerequisite for regular, stable fruiting process. Besides environmental factors, recent molecular studies have shown the role of genetic factors that control normal fruiting body production of Cordyceps. Recent progress in our understanding of Cordyceps fruiting in laboratory condition is discussed with some prospect of better fruiting body production of Cordyceps spp. in future.

Volvariella volvacea is a popular variety among people because of its distinct flavor, pleasant tastes, higher protein content and shorter cropping duration compared to other cultivated mushrooms. It is also the most expensive mushroom in artificial mushrooms. With the development of economy and the industrialization of Edible Fungi, more and more straw mushroom was grown in northern China, southern mushroom began move to northern. In this study, the author, based on many years of cultivation experience, summarizes the high yield cultivating techniques of Volvariella volvacea. Volvariella volvacea cultivation consists of five steps, the five steps are composting, pasteurization, spawning, pining and harvesting.

This study was carried out to investigate the management characteristics and techniques by comparing and analyzing the process of mixing, sterilization, cooling, inoculation and incubation for bag cultivation of L. edodes. The medium was sterilized after mixing it for 30 minutes, 1 and 2 hours respectively. The general moisture content of the sawdust substrate has been adjusted to 65%, while the case of the L. edodes substrate was 55%. Only 5% of wood chips of 5-8 mm in particle size were mixed to secure the space of the sawdust particles. As a result, mycelial growth was 1.4cm faster and the density was better(+++) than control(++). Wood chips are soaked in advance for a week during winter and 4 days during summer. There is an average number of 1.6x108 (cfu) of microorganisms in the sawdust that has been piled for six months outdoor. In summer, it has to be used immediately after mixing sawdust. High-pressure sterilization should be performed to use as a mushroom spawn, and to improve physical properties, it was great to sterilize the medium at a normal-pressure. There are height and width type for bags to be consumed for bagging. When the height was reduced into 17cm and the width was increased into 13cm, the browning period was shortened by 30 days and the period of mycelial growth was shortened by 25 days. The sterilized medium had an average of 4x103(cfu) of microorganisms at 25°C and 1.3×102 (cfu) at 15°C. After 25 days from inoculation in vitro, the growth condition of sawdust was the best with 13.2 cm, followed by grain spawn and liquid respectively. When inoculated with liquid spawn, the moisture content of substrate should be adjusted to 55% to 50% in advance.

For centuries, Lentinula edodes called “Pyogo” in Korea has been cultivated as a high-grade edible mushroom in Asia. This is known as medicinal mushroom for cold prevention in oriental medicine. In the world market, this mushroom of Lentinula edodes is introduced as representative edible and medicinal mushroom in Asia. As consumer interest increases, Asian Lentinula edodes market is also growing. There is a record that Korea has already been used as a gourmet food on the royal meal table for the king in the 15th century Joseon Dynasty. In Korea's famous oriental book, it is cooked with many traditional foods because of the pharmaceutical efficacy of this mushroom. For long time ago, difference in cultivation as well as in mushroom growth of Lentinula edodes has studied in Korea. Recent advances in cultivation of selected high quality mushroom studied from substrate optimization using log and sawdust of oak tree. Cultivation of Lentinula edodes strain research controlled the production of large quantity of various substrate conditions. This mushroom mycelia and fruit-body were cultivated in log of oak tree and sawdust substrate cultivation in types of bioreactors. According to compared cultivation method, it presented the optimistic condition for the amount production of various Lentinula edodes strain in Korea.

In order to examine the hypolipidemic and hepatic steatosis preventing activities, we investigated mechanism of Auricularia auricula-judae 70% ethanol extract (AAE) in vivo and in vitro. Normal diet (ND) and high fat diet (HFD) with or without 0.1% (w/w), 0.3% (w/w) and 1% (w/w) AAE were given to male C57BL/6 mice. Plasma lipids and liver enzymes were measured and tissue sections of liver were examined. Further mechanistic studies in mouse 3T3-L1 adipocytes were performed in vitro by verifying triglyceride, glycerol, GPDH (glycerol-3-phosphate dehydrogenase) activity and mRNA expression of adipogenic and lipogenic genes using RT-PCR amplification. Body weight and adipose tissue mass were significantly reduced in ND and (HFD plus AAE) fed mice compared to HFD mice. These findings suggest that AAE may reduce the risk of hepatic steatosis by modulating plasma lipids via the regulation of adipogenic/lipogenic transcriptional factors. AAE may have interesting applications to improve plasma lipids and liver enzymes (A part of results have been published in Int J Med Mushrooms).

Lignosus rhinocerotis, commonly known as Tiger Milk Mushroom, has been long extolled for its medicinal properties and used for treatment of asthma, cough, fever, cancer, liver-related illnesses, joint pains and as tonic. The history of usage for Tiger Milk Mushroom dates back to almost 400 years, ago but there were no records of scientific studies done due to inavailability of sufficient samples. Even when there were samples collected from the wild, the supply and quality was inconsistent. With the advent of cultivation success of one of the most utilized species of Tiger milk mushroom (L. rhinocerotis) in 2009 (known as TM02), scientific investigation was done to validate its traditional use and to investigate its safety for consumption, biochemical and biopharmacological properties. Preclinical toxicology evaluations showed that TM02 did not induce adverse effects on fertility. It also poses neither teratogenic effects nor genotoxicity. The no-observed-adverse-effect level (NOAEL) dose of the TM02 in 180-day chronic toxicity study is 1000 mg/kg which is equivalent to 162 mg/kg human dose, anticipated dose for a 60 kg adult is ~10g. The effective dosage of TM02 is as low as 0.5g per day for an adult. There is therefore, a huge margin for the safe consumption of TM02. TM02 is proven to be effective in supporting the lung and respiratory health, immune health, maintenance of joint health (caused by inflammation), improving vitality and stamina and improving the quality of life for cancer patients. Recent scientific findings have shown that TM02 contains various active components such as the polysaccharides-protein complexes, β-glucans, proteins, immuno-modulatory proteins all of which could play possible roles in rendering significant therapeutic properties such as anti-inflammatory, immuno-modulatory, anti-proliferative, anti-oxidative, neurite outgrowth stimulation etc. The revelation of the genome, transcriptome, proteome of L. rhinocerotis by MMRG has provided valuable insights into the biomolecule discovery and provided the foundation for future research and exploitation of L. rhinocerotis in pharmacological and functional food applications. These data forms a valuable foundation for future research in the exploitation of the L. rhinocerotis in pharmacological and industrial applications.

Lentinus tigrinus (L. tigrinus), a white-rot fungus that grows naturally on rotten hardwood during spring and summer in China, is an edible and medicinal mushroom containing a valuable combination of nutrients including high amino acid concentrations and low sugar levels. However, no reports have isolated and characterized FIP genes from L. tigrinus to date. In our study, two novel fungal immunomodulatory proteins (FIPs) from Lentinus tigrinus were identified and named Fip-lti1 and Fip-lti2. The bioactive characteristics of Fip-lti1 and Fip-lti2 were compared to a well-known FIP (LZ-8 from Ganoderma lucidum) to investigate the effect of Fip-lti1 and Fip-lti2 expression on Concanavalin A (ConA)-induced liver injury. Both Fip-lti1 and Fip-lti2 protected livers from ConA-induced necrosis, as evidenced by decreased serum aminotransferase levels (AST, ALT) and relieved liver histology. Levels of proinflammatory cytokines (TNF-α, IL-1β, and IL-6) and oxidative stress (SOD, MDA) were shown to be reduced by expressing Fip-lti1 and Fip-lti2. In addition, the hepatoprotective effect of Fip-lti1, Fip-lti2, and LZ-8 correlated with ameliorating the imbalance of Th1/Th2 (IFN-γ/IL-4). The observed liver protection of Fip-lti1 and Fip-lti2 was mechanistically explored. Treatments with Fip-lti1 and Fip-lti2 regulated GATA3/T-bet expression, activated the decreased Nrf-2/HO-1 pathway, and countered the upregulated NLRP3/ASC/NF-κBp65 signaling in ConA-stimulated liver injury. In conclusion, we identified two fungal proteins (Fip-lti1 and Fip-lti2) that can protect liver from ConA-induced liver injury.

The fungal mycelium-based composite is a new biomaterial to replace the existing composite material. To compete with lightweight, high-performance composites represented by fiber-reinforced plastic, various physical and chemical properties and functionality must be secured. Especially, the composite material made by using mushroom mycelium (or fruting bodies) is called mushroom plastic. Currently, Ecovative, Mycoworks, and Muskin in USA and Europe are launching new products including structural materials and leathers. Products utilizing mushroom mycelium can be launched in the market for construction materials, automobile interior materials and artificial leather substitutes. In spite of this high possibility, mass production using FMBC has not yet been reported. This presentation introduces the mycelium-based materials, a material that can replace existing plastics, inorganic building materials and animal skins in an environmentally and economically viable way, and looks at the possibility of future biomaterials by summarizing recent research contents.

Polygonum multiflorum (Polygonaceae family) is effective to cholesterol lowering effect, increased immunity effect and whitening effect. P. multiflorum root is used for medicinal herb that can be used as raw material for food. In this study, acetic acid fermentation was adding extracts of P. multiflorum root(0%, 1%, 3%, 5%) for the developing healthy beverage. In this study, we investigated the contents of free sugars, organic acids and inorganic components of P. multiflorum root. As a result of free sugars from P. multiflorum root analysis, three kind of free sugars were determined the glucose, sucrose and lactose by HPLC. Four kind of organic acids were found in lactic acid fermented P. multiflorum root. And oxalic acid, lactic acid and succinic acid were detected in organic acids. The content of lactose and lactic acid in lactic acid fermented P. multiflorum root was the highest among the free sugars and organic acids. The content of minerals were in high orders of sodium> calcium> magnesium in P. multiflorum by acetic acid fermentation. The content of inorganic components was highest in lactic acid fermented solution with 5% and proportional to the content of P. multiflorum

Oyster mushrooms are popular and physiologically beneficial to human health. This study was conducted to investigate the effect of dietary supplementation of oyster mushrooms such as Pleurotus ostreatus, P. pulmonarius, and P. citrinopileatus on high cholesterol fed rats. Five-week old female Sprague-Dawley albino rats were divided into three groups of 8 rats each. Feeding a diet containing a 5% fruiting body powder of Pleurotus species to high cholesterol fed rats reduced plasma total cholesterol (TG), triglyceride (TG), low-density lipoprotein (LDL). Feeding each oyster mushroom also significantly reduced body weight in high cholesterol fed rats. However, it had no adverse effects on plasma albumin, creatinine, blood urea nitrogen, uric acid, glucose, total protein or enzyme profiles such as glutamate oxalate transaminase (GOT), glutamate pyruvate transaminase (GPT), and alkaline phosphatase (ALP) which are related to kidney function. Feeding each mushroom increased total lipid and cholesterol excretion in feces. A histological study of hepatic cells by conventional red O staining showed normal findings in mushroom-fed hyperlipidemic rats. These results suggested that diet supplementation of 5% fruiting body power of Pleurotus species provided health benefits by acting on the atherogenic lipid profile in high cholesterol fed rats.

Occurrence spontaneous mushrooms in eastern Jeju Muryeongarioreum, Sillyecheon-Iseungak, Kyoraegotjawal, Geomunoreum were examined in 2016. The collected mushrooms were classified by 15 order 56 family 149 genera 317 species. Among the collected mushrooms, 87 species of mushrooms were successfully incubated in mushroom complete medium in which DNA of 25 species were completely extracted and amplified using universal primers ITS1 (5’-CTTG GTCA TTTA GAGG AAGT AA-3’) and ITS4 (5’-TCCT CCGC TTAT TGAT ATGC-3’). Four species of them were differently identified between the classification by morphological observation and by analysis of DNA sequences i.e. by morphological identification Ganoderma applanatum, Armillaria gallica, Macrolepiota procera and Trametes gibbosa were identified as G. gibbosum, A. cepistipes, M. detersa and T. elegans by analysis of DNA sequences, respectively. Based on these results both identification ways might be necessary for correct classification of mushrooms. On the other hand, among the collected 317 mushrooms, 48 species could be identified to genus but not to species by the morphological observations. However, by the analysis of DNA sequences, 15 species were identified to species in which 6 species were unregistered mushrooms in Korea. Therefore, the analysis of DNA sequence may be useful to identify the mushrooms which are difficult to recognize by morphological observation.

Amanita Pers. is a well-known monophyletic mushroom genus with a broad distribution. The genus comprises about 500 named taxa and contains deadly poisonous species, e.g. A. virosa and A. phalloides. Nonetheless, the diversity of Korean Amanita species has been underestimated. At present ca. 50 species of genus Amanita have been reported and distributed in Korea. It caused mainly three problems: 1) identification is not clarify because of the morphological investigation only; 2) poor morphological descriptions of the species; 3) absence of evidence specimens in many case of the recorded Korean Amanita species. For these reasons, the goals of the present study were to establish the accurate classification systems and to understand the diversity of Korean Amanita species. Macro-morphological characters were determined based on structures of basidiomata (universal veil on the pileus, partial veil, volva etc.), field notes and color photos of basidiocarps. Micro-morphological characters (basidiospores, basidia, composition of volval remnants on pileus and stipe base etc.) were obtained from the dried specimens after sectioning and rehydration using optical microscope. For the molecular identification, four gene regions(nrITS, nrLSU, rpb2 and tef1) were sequenced from Korean Amanita species. Among the sequenced specimens, I selected 80 specimens (which were successfully sequenced as ITS, nLSU, rpb2 and tef1) to generate the outline of Amanita phylogeny including 2 subgenera and 7 sections for determination of exact phylogenetic positions and comparison between related species. As a result of 11 data sets of each subgenus and complexes, 68 species were represented including new to Korea and candidates of new to sciences. The protein coding genes were helpful to determine the grouping within a related species and/or complex groups rather than rDNA sequences regions.