Hypsizigus marmoreus is commercially the most important edible mushroom in Japan. This mushroom is usually cultivatedfor a longer period (about 85~120 days) than other mushroom. In order to develop a new cultivar that has a shortened cultivationperiod, the genome analysis of this strain has been considered. This study aims to obtain parental monokaryotic strains reproducing‘Haemi’ cultivar in Hypsizigus marmoreus for reference genome sequencing. The mycelia were cultured in MCM and MYG media forvarious incubation periods. Homogenized mycelia were treated with commercial cell wall degrading enzymes to maximize protoplastsproduction yield from Hypsizigus marmoreus. The greatest number of protoplasts was obtained from mycelia cultured in MCM mediafor 3 days using Novozyme enzyme. The isolated protoplasts were grown in regeneration agar media after two weeks. Regeneratedcolonies were picked and moved on separated dishes for microscopic observation. Neohaplonts regenerated from dikayotic strainswere identified by the absence of clamp connections. We confirmed that one of monokaryotic strains is a parental strain by crossingwith an original compatible strain of ‘Haemi’ cultivar. This parental strain will be used for reference genome sequence analysis.

Agaricus bitorquis is an edible white mushroom of the genus that is cultivated at high temperature(25±1oC) unlikeA. bisporus is cultivate at 16±2oC. Unlike Agaricus bisporus, an edible white A. bitorquis mushroom is cultivated at hightemperature (25±1oC). Most farmers cultivate this mushroom for a long cultivation period in Korea. For this reason, we madeheterokayons to develop a new cultivar that generate fruitbodies for short cultivation period. Over one hundred SSIs(singlespores isolates) were collected from selected A. bitorquis ASI1151 and ASI1349 strains. Seventy-three SSIs were germinated onCDA(compost dextrose agar) media after 20 days (minimum) or 83 days (maximum) incubation under different mediacondition. The mycelial growth rate of germinated SSIs was different. 9 homokaryons in ASI 1151 and 11 homokaryons in ASI1349 from SSIs were selected by OPN-02 primer in RAPD analaysis. Also this primer was used to select heterokaryon thatcross among each homokaryon with compatible locus. Therefore 44 compatible matings were confirmed of 99 crossed lines.

A new brown button mushroom cultivar, ‘Hogam’, C34 line, was made by crossing homokaryons, ASI1164-37 andASI1175-66, selected by RAPD analysis and by cultivating three times. Mycelium of ‘Hogam’ on CDA (compost dextrose agar)media grew well at 25oC. The optimum pin-heading temperature of new variety and optimum growing temperature was 14-18oC. The thickness of the mature cap and stipe were thicker than a control, ‘Dahyang’ that developed in 2010. The color ofpileus was light brown and lighter than ‘Dahyang’. Days required from casing to first harvesting were three days longer thancontrol strain, but the weight of harvested fruiting body increased by 1.35 times. ‘Hogam‘ cultivar are expected to contribute tothe diversification of domestic mushroom cultivars.

A new commercial strain “Mongdol” of oyster mushroom was developed by hyphal anastomosis. It was improvedwith hybridization between monokaryotic strain derived from Pleurotus ostreatus ASI 0627 and dikaryotic strain derived from P.ostreatus ASI 2929. The optimum temperature of mycelial growth and fruiting body development were 25~30oC and 12~18oC,respectively. When two different media including PDA (potato dextrose agar medium) and MCM (mushroom complete medium)were compared, the mycelial growth of this mushroom was faster in MCM than in PDA. Similar result was observed with thecontrol strain P. ostreatus ASI 2504. Analysis of the genetic characteristics of the new cultivar “Mongdol” showed a different DNAprofile as that of the control strain ASI 2504, when RAPD (Random Amplified Polymorphic DNA) primer URP3 and URP6 wereused. Fruiting body production per bottle was about 106 g using demonstration farms. The color of pileus was blackish gray andthe stipe was long. Therefore, we expect that this new strain “Mongdol” will satisfy the consumer’s demand for high qualitymushrooms.

‘Baekjung’ adaptable to high temperature was made by crossing between monokaryon derived from selfing of brownstrain and monokaryon derived from Korea white strain. In the condition that temperature is maintained at 10oC without lowtemperature of 4oC suppressing treatment and wrapping during cultivation period, it showed good productivity thanUri1ho(control). The optimum temperature of mycelial growth was 30oC and that of fruiting body initiation and developmentwere 14oC and 7oC, respectively. The days for the fruiting and yield were 7days and 277±11.2g per 1,100ml bottle,respectively. This variety needed high concentration of carbon dioxide up to 4,000 ppm for the good quality.

The white button mushroom, Agaricus biporus is commercially the fifth important edible mushroom, accounting for a Korean production of 10,996 tons in Korea, 2012. The button mushroom, A. bisporus, mostly cultivated at mushroom farm controlled 16-19°C temperature during fruiting period in all seasons. For this reason, the production costs including the cost of energy, is very expensive to keep optimal culturing temperature. In this study, mycelial growth of strains collected from various countries was investigated at different cultivation temperature conditions to use baseline data for developing thermo-tolerance varieties. Mycelial growth was recorded strains cultivated in CDA (Compost Dextrose Agar) after two weeks. 12 strains among total 264 strains were cultivated well in high temperature. These strains will be cultivated in high temperature to confirm relationships between mycelial growth and fruit-body formation in same conditions.

This study was carried out to investigate neuronal protective activity of fruiting body of Hericium erinaceum. In order to search the effective active compound against amyloid beta peptide-induced oxidative stress on neuronal cells, rat pheochromocytoma cells (PC12), Extracts of Hericium erinaceum were screened and evaluated using both the 2’,7’-dichlorofluorescin diacetate assay (DCF-DA), and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. According to above assays, Solvent partitions of extracts were selected for further purification and isolation of anti-Alzheimer’s disease compound as it exerted the highest protective effects against hydrogen peroxide (H2O2)-induced oxidative stress.

Many basidiomycetes, especially mushroom species, thrive on saccharides that they obtain from the decomposition of cellulose, hemi-cellulose and or lignin. Cellulose is degraded by specific enzymes called cellulases. Cellulases play an important role in the biosphere by recycling cellulose, the most abundant carbohydrate produced by plants. Also cellulases have many potential biotechnologic and industrial applications. Detection of cellulose degrading activity is commonly performed on carboxymethylcellulose CMC medium in combination with a stain that reveals the degraded CMC. In order to efficiently screen the F. velutipes knockout library, obtained through Agrobacterium-mediated transformation (ATMT), for those important enzymes we compared two different staining methods i.e. Congo Red and Gram’s Iodine (as reported in R. C. Kasana et al 2008). The latter stain showed strongly enhanced detection in time and intensity, facilitating mass screening of our mutant database. Using Gram’s iodine and square petri dishes containing up to 9 colonies at a time we can now rapidly screen multiple mutants in a short period. We are going to find the genes. Inactivated genes of mutants with altered cellulase degradation activity will be identified and cloned for further analysis.

We investigated 18 amino acid contents in the fruiting bodies of Agaricus strains to understand their variation in different growth temperature. Although total content of amino acid was more or less different depending on the isolates, it increased until the temperature was 22℃ and decreased above 22℃. Commercial strains of A. bisporus, such as Yangsongi-505, Yangsongi-705 and commercial strain A collected in the market exhibited these general trend in total amino acid content. However, another commercial strain B did not follow these trend. A strain of A. bitorquis did not produce mushrooms at 10 and 13℃, but mushrooms harvested in the other treated temperatures exhibited with the similar patten. Generally, the most amino acid contained in Agaricus spp. was cysteine and followed by phenylalanine, glutamic acid, lysine, proline and histidine. We detected two trends for the contents of each amino acid in the fruiting bodies of Agaricus spp. grown at the different growing temperature. Some amino acid contents were largely correlated with the growth temperature, but the others showed the trend of increase until 22℃ and decrease above 22℃.

We investigated 18 amino acid contents in the fruiting bodies of Agaricus strains to understand their variation in the harvesting periods. Total content of amino acid was different depending on the isolates. In the case of A. bitorquis and A. brunnescens, the total amino acid content increased and correlated with the harvesting periods. For commercial strains of A. bisporus, such as Yangsongi-505, Yangsongi-705 and commercial strain A collected in the market, it increased until the second harvesting period, but decreased in the third harvesting period indicating that these three isolates exhibit the exceptions for the general trend. Among isolates we used in our experiments, the isolate of A. brunnescens contained the highest and two isolates of A. bisporus were the lowest in total amino acids. We found two trends for the contents of each amino acid in the fruiting bodies of Agaricus spp.. One is the same with the trend of the total amino acid content and the amino acid content were largely correlated with the harvesting period. The other is the trend of increase of amino acid until the second harvesting period and decrease of amino acid in the third harvesting period, which was also shown in the total amino acid content of A. bisporus. Generally, the most amino acid contained in Agaricus was cysteine and followed by phenylalanine, glutamic acid, lysine, proline and histidine.

Hypsizygus marmoreus, belongs to Tricholomataceae of Agaricales, is widely cultured as the Beech mushroom in Japan. “Haemi” is the first variety developed by intra-specific crossing in Korea. It was improved with hybridization between monokaryotic strain derived from MKACC51987-8 and MKACC51988- 12. The optimum temperature of mycelial growth and fruiting body development were 23-25℃ and 11~17℃, respectively. The color of fruitingbody was grayish brown and cap type was umbrella. It suggest that 'Haemi' is new commercial variety for consumer who concern over healthy and tasty food

Flammulina velutipes, amongst others known as Winter Mushroom or Enokitake is an important economic crop in Asia. The tetrapolarity (having four mating types) of this mushroom obligates mating and results in self-sterile progeny that carries unique genetic traits, making understanding of the genetic base desirable for breeding. Moreover, mating type genes are significant for evolutionary studies as their high polymorphism benefits phylogenetic comparisons. This polymorphism further makes mating type genes interesting candidates for genetic markers that allow identification of specific strains. Mating type loci in Agaricomycotina are classically termed A and B and control two different developmental pathways [for a review see 1]. They consist of tightly linked subloci that encode multiallelic genes. MatA loci contain two groups of divergently transcribed homeodomain proteins (HD1 and HD2) and heterodimerization of HD1 with a non-self HD2 protein forms a functional transcription factor that activates the A pathway. MatB loci hold pheromones and pheromone receptors. Pheromone genes encode small precursor proteins that are characterized by a C-terminal CAAX motif. Pheromone receptors typically contain 7 membrane spanning regions and are coupled to G-proteins. Binding of a pheromone to a receptor, triggers splicing of the (trimeric) G-protein, which activates the B pathway. New genetic data from recent genome sequences is challenging the strict concepts of old mating type models in fungi. MatB loci turn out to be rather diverse and contain considerable varying numbers of pheromone receptors and associated pheromones. To this, pheromone receptors which are not linked to matB loci have now been reported for C. cincerea, S. commune and L. bicolor [2, 3]. Also the organization of the matA locus is less strictly conserved than anticipated. Though far more tightly maintained than the matB locus, substantial differences in HD gene numbers and overall organization are reported [2, 3]. These differences stress the importance of determination of the individual mating type systems from industrially important mushrooms to assist breeding. Our analysis of F. velutipes strain 4019-20 uncovered 7 pheromone receptors together with 3 pheromones. The matB-3 locus of this strain however, is defined by only a single pheromone receptor and pheromone gene and our data strongly indicates that a 2nd pheromone receptor recently lost its function. The other receptor genes are non mating type specific. Finally, we detected three homeodomain genes distributed over two distant subloci. These subloci have been separated by two large inversions. Strikingly the distant matA subloci in S. commune seem to be separated by inversions as well. Synthenic mapping of a large regions from Coprinus cinerea, Laccaria bicolor, S. commune and F. velutipes shows that the matA loci originate from a single locus in a common ancestor of S. commune and F. velutipes that is represented by L. bicolor and C. cinerea. [1] U Kües. Micr Mol Biol Rev 64, 316 (2000) [2] H Niculita-Hirzel, J Labbé, A Kohler, F le Tacon, F Martin, IR Sanders and U Kües. New Phytol 180, 329 (2008). [3] RA Ohm, JF de Jong, LG Lugones, A aerts, E Kothe, JE Stajich, RP de Vries, E Record, A Levasseur, SE Baker, KA Bartholomew, PM Couthino, S Erdmann, TJ Fowler, AC Gathman, V Lombard, B Henrissat, N Knabe, U Kües, WW Lilly, E Lindquist, S Lucas, JK Magnuson, F Piumi, M Rausdaskoski, A Salamov, J Schmutz, FWMr Schwarze, PA van Kuyk, JS Horton, IV Grigoriev and HAB Wösten. Nat. Biotechnol ISSN: 1087-0156 (2010).

This study was carried out to investigate characteristic pattern of fruiting body of Ganoderma lucidum and their antioxidant activity. Mycelia of all strains were firstly inoculated into potato dextrose agar(PDA) and then transfered to a media of saw dust which contained 20% rice bran. These mycelia of saw dust were then inoculated into oak tree in polyethylene bags which has been sterilized for 8h at 120℃. The polyethylene bags were sent to a growth room for growth of fruit bodies. Antioxidant activities of each fruiting body were examined by using DPPH(α,α-diphenyl-β-picrylhydrazyl).

This study was carried out to investigate amino acid contents of golden mushroom and pink mushroom. The amino acid analysis was followed by AccQ-Tag method and HPLC on gradient conditions. Seventeen amino acids were analyzed and sixteen amino acids were found in golden mushroom ; fifteen amino acids in pink mushroom respectively. Among total amino acid in golden mushroom, cystein content was the highest and glycine, glutamic acid, proline were followed. Among total amino acid in pink mushroom, cystein was the highest and glycine, lysine, methionine were followed. As shown in Fig.1, 2, concerning amino acids, cystein was dominant. and alanine was detected in golden mushroom and Pleurotus ostreatus sold in market but pink mushroom. Phenylalanine was detected in Pleurotus ostreatus sold in market but that was not detected in golden mushroom and pink mushroom.

주로 고온에서 재배되는 왕송이는 온도제어기를 사용하지 않고 재배할 수 있는 잇점이 있다. 여름철 냉방시설없이 재배할 수 있는 품종을 개발하고자 국내외에서 수집된 왕송이균을 교잡하여 새로운 품종 ‘백련’을 육성하였다. 백련 왕송이의 균배양온도는 25℃~30℃이며 자실체 발생온도와 생육온도 거의 동일하다. 다만 배양완료 후 복토를 해야하기 때문에 2차 균사생장기간이 있어 초발이소요일수가 다소 늦다. 하지만 복토를 하기 때문에 주야간 온도차이에도 버섯발생이 용이하고 습도관리가 쉬워 여름철 재배가 유리하다. 버섯의 모양은 순백색에 전형적인 우산형으로 강한 다발성을 가지며 대가 긴편이다. 특히, 병재배보다는 봉지재배를 할 경우 다발성은 더욱 강해지면 자실체의 크기도 증가한다. 적정배지는 포플라 40%+미송 40%+미강 20%를 혼합한 배지이며 평균수량은 병재배시 101g이나 봉지재배에서는 더욱 증가할 것으로 사료된다.