This study was conducted to diversify the cultivation of mushroom items and develop a competitive variety of Hypsizigus marmoreus. We focused on developing Hypsizigus marmoreus with lower bitterness, likable shape, and high yield. We have collected and tested characteristics of genetic resources from domestic and abroad since 2019. Breeding (2019), characterization (2020), productivity test (2021), and farm demonstration test (2021) have been sequentially conducted. We bred the new variety ‘Yunseul’ with unique traits. The optimal temperature for mycelial and fruit body growth were 22–25oC and 15– 18oC, respectively. It was similar to the control variety (Mangadak-2Ho) in the pileus form (hemispherical shape) and the cultivation period. However, it was thinner and longer than the control variety with the pileus diameter, thickness, stipe diameter, and length being 19.9 mm, 7.0 mm, 9.4 mm, and 86.3 mm, respectively. The effective number of fruit bodies was 47.8 in bottle cultivation, which was more than that of the control variety. The yield was 197.4 g/bottle (1,100 cc), which was 30% higher than the yield of the control variety, 151.9 g/bottle (1,100 cc). The parent and control varieties were also incubated alongside the new variety. The somatic incompatibility line was distinct. The band pattern in the mycelial DNA PCR reaction was different from that of the parent and control varieties, confirming the hybrid species.

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.

A new commercial cultivar ‘GARES-HM-17-1’ of Hypsizigus marmoreus was developed by the method of mono-mono crossing between monokaryotic strains derived from Hae-Sal 3Ho. The parental strains are characterized by the property of high quality and a short period of time in cultivation. The optimum temperature of mycelial growth was 25°C and that of fruiting body development was 15~16°C. The period of harvesting including primordia formation was 23.0 days shorter than that of control strain Mangadak No. 2. The color of pileus and stipe surface was neutral-white, respectively. The yield was 82.0±5.8g/850cc plastic bottle. Analysis of the genetic characteristics of the new commercial cultivar ‘GARES-HM-17-1’ showed a different profile as that of the control strain, Mangadak No.2, when RAPD (Random Amplified Polymorphic DNA) primer was used. This new cultivar ‘GARES-HM-17-1’ of H. marmoreus is characterized by the improved quality and a short period of time in cultivation after scratching. It would be of much help to expand the cultivation area of H. marmoreus.

A new commercial cultivar ‘GARES-HM-16-1’ of Hypsizigus marmoreus was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR8047 and KNR8080. The parental strains are characterized by the property of high quality and a short period of time in cultivation respectively. The optimum temperature of mycelial growth was 25°C and that of fruiting body development was 15~16°C. The period of harvesting including primordia formation was 20.0 days shorter than that of control strain Mangadak No.2. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The yield was 123.7±14.8g/850cc plastic bottle. Analysis of the genetic characteristics of the new commercial cultivar ‘GARES-HM-16-1’ showed a different profile as that of the control strain, Mangadak No.2, when RAPD (Random Amplified Polymorphic DNA) primer was used. This new cultivar ‘GARES-HM-16-1’ of Hypsizigus marmoreus is characterized by the improved quality and a short period of time in cultivation after scratching. It would be of much help to expand the cultivation area of Hypsizigus marmoreus.

A new commercial cultivar ‘Hae Sal’ of Hypsizigus marmoreus was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR8047 and KNR8080. The parental strains are characterized by the property of high quality and a short period of time in cultivation respectively. The optimum temperature of mycelial growth was 25°C and that of fruiting body development was 15～16°C. The period of harvesting including primordia formation was 7.0 days shorter than that of control strain ‘Mangadak No.2’. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The yield was 117.6±23.2g/850cc plastic bottle. Analysis of the genetic characteristics of the new commercial cultivar ‘Hae Sal’ showed a different profile as that of the control strain, ‘Mangadak No.2’, when RAPD (Random Amplified Polymorphic DNA) primer was used. This new cultivar ‘Hae Sal’ of Hypsizigus marmoreus is characterized by the improved quality and a short period of time in cultivation after scratching. It would be of much help to expand the cultivation area of Hypsizigus marmoreus.

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.

전 세계적으로 최근 아토피 피부염의 유병율이 증가하면서 이 문제에 대한 관심이 증가하고 있지만 근본적으로 개선시킬 수 있는 효과적인 치료법이 개발되지 못한 상태로 단순히 증상을 경감시키는 대증요법인 스테로이드 같은 약물치료만이 현재 표준 치료로 시행되고 있는 실정이다. 이러한 약물치료는 장기적인 측면에서 내성 증가와 증상의 악화 등의 문제점으로 인해 최근에는 천연성분을 이용한 항아토피 소재에 대한 연구들이 활발히 진행되고 있다. 그 중 식용버섯인 Hypsizigus marmoreus(HM, 느티만가닥 버섯)는 많은 생리활성 물질을 포함하고 있으며 특히, 항염효과가 우수한 것으로 밝혀져 있는데 본 연구에서는 HM이 아토피 피부염에 미치는 효과를 측정하였다. 동결건조한 버섯 시료를 85% 에탄올로 추출하여 추출물을 제조하였으며, 이를 동결건조하여 시료로 사용하였다. 마우스 유래의 대식세포인 RAW264.7에 염증 유발 물질인 LPS(lipopolysaccharide)와 HM 추출물을 처리하여 염증 관련 싸이토카인 TNF-α, IL-6, IL-2, IL-4 및 IFN-γ과 nitric oxide(NO)생성량을 확인하였다. RAW264.7 세포주에 HM 추출물을 농도별로 처리한 결과, 고농도로 처리된에서 높은 NO 생성 저해능을 보여주었다.

아토피 피부염은 소양증, 가려움, 각질형성, 홍반 등을 동반하며 현대인의 삶의 질에 큰 영향을 미치고 있는 질환으로 전세계적으로 아토피 피부염 환자가 급격하게 증가하는 추세이다. 국내의 경우에도 아토피피부염으로 인한 진료 인원이 연간 100만명 이상 집계될 정도로 심각한 보건사회문제로 대두되고 있지만 아직까지 아토피 피부염에 대한 근본적인 치료법이 개발되지 못한 상황이다. 단순히 증세 완화를 위해 피부보습 유지와 염증억제 등의 방법을 일반적으로 사용하고 있지만 과도한 보습제 사용이나 증상 경감을 위한 스테로이드와 같은 약물처방은 더욱 증상 악화시키며 장기간 사용시 내성 증가 등의 악순환으로 이어질 수 있다. 최근 이러한 근본적인 치료를 위한 해결 방법으로 천연 물질로부터 아토피 피부염을 경감시킬 수 있는 소재의 연구가 이루어지고 있다. 본 연구는 다양한 생리활성과 함께 항염 효과가 우수한 것으로 알려진 식용 버섯인 Hypsizigus marmoreus(느티만가닥 버섯) 추출물의 항 아토피 활성과 Hypsizigus marmoreus 추출물이 개별인정형 피부보습 개선 소재인 식물복합추출물(AF-343)을 함께 사용시 아토피피부염 개선효과에 미치는 영향을 알아보기 위해 실시하였다. 동결건조한 Hypsizigus marmoreus를 85% 에탄올로 추출하여 제조한 추출물과 AF-343을 사용하여 사료로 제작하여 실험에 사용하였다. 제모한 6주령의 Balb/c 마우스의 등과 귀부분에 5% picryl chloride(PCL)를 도포하여 아토피 피부염을 유발시킨 후, 5주 동안 Hypsizigus marmoreus 추출물과 AF-343으로 제조한 사료를 Balb/c 마우스에 급여하면서 변화를 관찰하였다. 실험군은 무처리 정상군, PCL만 도포한 대조군, 아토피 치료제 도포군(1% 히드로코르티손)과 Hypsizigus marmoreus 추출물 급여군 및 Hypsizigus marmoreus 추출물과 AF-343의 동량 혼합물 급여군으로 진행하였으며 5주간의 사료 급여 후 희생시켜 등조직의 RT-PCR을 실시하였다. Hypsizigus marmoreus 추출물 단독 급여군과 AF-343을 혼합하여 급여한 군은 아토피 피부염이 유발된 Balb/c 마우스에서 PCL만 처리한 대조군에 비해 피부의 발진이 감소되는 효과를 관찰 할 수 있었다. 또한, 마우스 등조직의 RT-PCR 결과에서도 Hypsizigus marmoreus 추출물 단독 급여군과 AF-343의 동량 혼합 급여군이 PCL 처리 대조군에 비해 염증성 인자인 COX-2와 iNOS 의 mRNA 발현양이 낮은 것으로 나타나 Hypsizigus marmoreus 추출물의 항아토피 활성과 피부보습 소재인 AF-343과의 혼합 사용으로 아토피 피부염을 억제하는 것으로 확인되었다.

본 연구는 느티만가닥버섯 20품종 버섯, 잿빛만가닥 3품종 버섯, 땅지만가닥 1품종 및 돌연변이 느티만가닥버섯 20종류의 유전적 변이를 RAPD 방법에 의해 분석하고자 수행하였다. 이들은 한국, 중국, 일본, 대만 등에서 유래한 것들이다. 느티만가닥버섯에 감마선을 조사하여 돌연변이체를 만들었다. 본 연구에 이용한 primer들은 40종류였고 이 중 31종류의 primer등이 반응을 나타내었다. RAPD 결과들을 분석한 결과 이 들 품종으로는 7개의 cluster로 나뉘어 졌다. 돌연변이체들은 유전적으로 유의적 차이가 있는 subgroup으로 나눌 수 있었다. 이상의 결과들은 본 실험에 사용한 primer들과 포자의 감마선 조사 등을 버섯의 신품종 개발에 유용한 도구가 될 것으로 생각된다.

The Hypsizigus marmoreus has been used as medicinal and food source in worldwide. Viridans Streptococci, which include Streptococcus sanguinis and Streptococcus gordonii, are pioneer oral bacteria that initiate dental plaque formation. This study was to investigate the antimicrobial activity of Hypsizigus marmoreus Extracts. Antimicrobial activity of extracts were tested against pathogenic bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus sanguinis and Streptococcus gordonii) by paper disc methods. The Hypsizigus marmoreus Extracts were extracted with 80% methanol. The 80% methanol extracts of Hypsizigus marmoreus Extracts showed antimicrobial activity against S. sanguinis and S. gordonii. The 80% methanol extract of Hypsizigus marmoreus Extracts were fractioned by hexane, chloroform, ethylacetate and buthanol. The ethylacetate fraction of Hypsizigus marmoreus Extracts showed strong antimicrobial activity against S. sanguinis and S. gordonii with minimum inhibitory concentration (MIC) range of 0.8-1 mg/ml. Especially, the 80% methanol extract and ethylacetate fraction of Hypsizigus marmoreus Extracts were inhibited the biofilm formation of S. sanguinis and S. gordonii at a concentration of 1.0 mg/ml. These results suggest that Hypsizigus marmoreus extracts can be developed as a potent antimicrobial agent.

The mushrooms have been used as traditional medicines and food resources in many countries. The objective of this study was to determine antioxidant compounds and to evoluate tyrosinase inhibitory activity of extracts from Hypsizigus marmoreus. The pileus and stipe of Hypsizigus marmoreus were extracted with methanol and water, separately. The methanol extract of Hypsizigus marmoreus were fractioned by hexane, chloroform, ethylacetate and buthanol. The concentrations of total polyphenolics and flavonoids in methanol extracts were investigated by colorimetric methods. The concentrations of total polyphenolics and flavonoids in methanol extract of the pileus was higher than methanol extract of the stipe. The DPPH redical scavenging activity of the pileus extract was also higher than stipe extract in methanol extract. The IC50 of DPPH redical scavenging activity of the pileus and stipe in methanol extract were 18 mg/ml and 1 10 mg/ml, respectively. The IC50 of tyrosinase inhibitory activity of the pileus and stipe in methanol extract were 500 mg/ml and 1,000 mg/ml in methanol extract. These results suggest that Hypsizigus marmoreus can be potentially used as a source of natural antioxidant agent in the cosmetic industry as well as the food, pharmaceutical and medicinal industry.

Recently, Hypsizigus marmoreus is noticed as new mushroom item in Korea. However, this mushroom has the long incubation period about 80 days including pre-incubation period. This study was attempted to establish the optimal post-incubation temperature and period of Hypsizigus marmoreus for bottle cultivation. The most effective post-incubation temperature and period were 25℃ and 20days, respectively. The higher yield was obtained under 25℃ for 20 day after 30 days of pre-incubation. Therefore, total cultivation period was shortened from 106 days (control) to 78days. It is useful information to increase rotation time of cultivation for effective management in H. marmoreus.

본 연구는 느티만가닥 버섯의 식품학적 이용성 증진을 위한 기초적인 연구로써 느티만가닥 버섯의 일반성분, 아미노산 및 휘발성 향기성분을 분석하였다. 느티만가닥 버섯의 일반성분은 탄수화물이 60.1%, 조단백질 32.0%, 수분 8.98%, 회분 5.0%, 조지방 2.0%의 함양을 보였다. 무기물함량은 건조물 100 g을 기준으로 칼륨 429.5 mg, 인 101.9 mg, 나트륨 20.3 mg, 마그네슘 54.86 mg, 칼슘 2.7 mg, 아연