Hypsizygus marmoreus is a mushroom with abundant flavor and medicinal properties. However, its application is limited by problems such as long cultivation period, low biological efficiency, and microbiological contamination; therefore, there is a substantial need for development of new cultivars of this species. In this study, 55 strains of H. marmoreus were subjected to inter simple sequence repeat (ISSR) analysis to identify markers for the selection of mother strains for breeding from the collected germplasm. ISSR 13 and 15 were confirmed as polymorphic markers. The three strains (KMCC03106, KMCC03107, and KMCC03108) with white cap color were found to be genetically closely related upon UPGMA analysis of both ISSR 13 and 15. Based on the PCR analysis results for ISSR 15, the collected germplasm were differentiated into three groups according to the strain collection year. Thus, ISSR 15 could be a marker for determining the phylogeny of cap color and genetic variations according to the strain collection year. These results suggest that ISSR markers can be effective tools for the selection of mother strains for breeding of H. marmoreus.

H. mamoreus is a mushroom with abundant favor and medicinal use. However, its cultivation has problems such as the long cultivation period, low biological efficiency and microbiological contamination, so new cultivars should be developed. In this study, 55 strains of H.marmoreus were analyzed with ISSR markers to identify precise genetic information in collected germplasm as mother strains in breeding. ISSR 13 and 15 markers were confirmed polymorphism. The three strains (KMC03106, KMC03107, and KMC03108) with white cap color were close genetic relationship in the UPGMA analysis of both the ISSR 13 and 15 markers. Especially in the PCR result of the ISSR 15, the collected germplasm were differentiated to three groups following collecting year. As these results, the ISSR 15 marker would be able to classify the phylogeny of cap color and genetic variation along the collecting year. Therefore ISSR markers will confer effective selection of mother strains for breeding of H. mamoreus.

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.

수집된 주름버섯류의 233점의 phylogenic relationship 을 분류하고 자실체형태를 조사하였다. 리보좀 DNA의 ITS 부분의 염기서열을 분석을 통해 38점이 다르게 동정 되었다. 수집된 주름버섯의 계통수에서는 양송이와 여름 양송이가 group A 내에서 주름버섯은 group B에서 가까 운 유연관계를 보였다. 또한 수집된 국가별 및 양송이내 갓색에 따른 유연관계가 있는 지 알아보았으나, 그 차이 는 없었다. 수집된 주름버섯류의 자실체 특성을 조사하기 위해 1차, 2차 두 번 재배를 하였다. 이 결과로 주름버섯 류 수집균주의 재배의 유무를 알 수 있었고, 버섯 종류와 시기에 따른 수확량 및 자실체의 특성을 비교를 통해 주 름버섯이 2차 재배시 수확량 및 경도에서 우수한 특성을 보이며, 양송이 중에는 비백색양송이가 단단하다는 것을 알 수 있었다. 또한, 여름양송이의 초발이 소요일수와 갈 색양송이의 갓 색이 외부환경에 영향을 받는 다는 것을 보여주었다.