한국에서 Mattirolomyces terfezioides와 Tricholoma bakamatsutake의 보고

가강현,전성민,유림,강정아,홍기성,Ka, Kang-Hyeon,Jeon, Sung-Min,Ryoo, Rhim,Kang, Jung-A,Hong, Ki-Sung 한국균학회 2015 韓國菌學會誌 Vol.43 No.2

상업적으로 중요한 균근성 버섯인 Mattirolomyces terfezioides와 Tricholoma bakamatsutake가 각각 한국의 아까시 나무와 신갈나무림에서 발견되었다. 이들 종은 형태학적 및 분자생물학적인 방법을 통해 동정되었으며, 두 종의 버섯에 대한 한글명도 각각 부여하였다. Mattirolomyces terfezioides and Tricholoma bakamatsutake, commercially important mycorrhizal mushrooms, were found for the first time in the forests of Robinia pseudoacacia and Quercus mongolica of the Korean peninsula, respectively. Morphological and molecular characteristics were discussed in the paper. We have also given the Korean name to the fungi here.

Panellus edulis 수집 균주의 균사배양 및 자실체 특성 분석

우성이 ( Sung-i Woo ),유림 ( Rhim Ryoo ),장영선 ( Yeongseon Jang ),박영애 ( Youngae Park ),정연석 ( Yeun Sug Jeong ),가강현 ( Kang-hyeon Ka ) 한국균학회 2018 韓國菌學會誌 Vol.46 No.3

Molecular analysis using the internal transcribed spacer region sequences revealed that the strains used in this study, which were formerly identified as Panellus serotinus, are Panellus edullis. After Universal Fungal PCR Fingerprinting (UFPF) analysis, eight strains of P. edulis were divided into two groups. We conducted fundamental research on mycelial growth and sawdust cultivation to understand the cultural characteristics of eight wild P. edulis strains collected from Korean forests. All strains showed faster and denser mycelial growth on potato dextrose agar (PDA) than on other media (malt extract agar, Sabouraud dextrose agar). Optimal conditions for mycelial growth were: 20℃ on PDA, 25℃ on potato dextrose broth (PDB), and pH 5~8 on PDB at 25℃. Two strains (NIFoS 2407, 3993) were selected as excellent strains based on mycelial growth and density on PDA. NIFoS 2792 showed high cellulase activities on carboxymethyl cellulose (CMC) agar, and NIFoS 2387 and 2804 exhibited high laccase activities on ABTS-containing agar media. The mycelial growth of P. edulis was the fastest on Quercus acutissima and Q. mongolica sawdust media, and mycelial density was the highest on Quercus spp. sawdust-containing media. Sawdust cultivation of P. edulis was successful. The conditions were 80~85 days of cultivation period after spawn inoculation, 10~11 days for primordial formation at 17~18°C, and 15~20 days for fruiting growth. NIFoS 2804 and 3993 were selected as good strains in terms of cultivation period and mushroom production. These results could be useful for the artificial cultivation of P. edulis.

표고 담자포자 유래 단핵균사의 A 교배형과 생장 속도 상관관계

박미정 ( Mi-jeong Park ),유림 ( Rhim Ryoo ),장영선 ( Yeongseon Jang ),가강현 ( Kang-hyeon Ka ) 한국균학회 2021 韓國菌學會誌 Vol.49 No.4

표고는 사극성의 교배계를 갖는 담자균의 일종으로, 표고의 교배형은 A와B라 불리는 서로 독립된 두 유전자좌에 의해 결정된다. 이론적으로 하나의 이핵균주는 네 개의 서로 다른 교배형을 갖는 담자포자를 1:1:1:1의 비율로 만들 수 있다. 과거 연구 결과에 따르면, 표고 담자포자에서 교배형이 편향된 분리비로 나타남이 보고되었다. 하지만 이러한 결과들은 꺽쇠연결과 같은 형태학적 특성만을 기반으로 교배형을 결정했다는 한계가 있다. 이에 본 연구에서는 교배형의 편향된 분리비가 표고에서 일반적인 현상인지 보다 명확하게 알아보기 위해서 최근에 보고된 DNA 마커를 활용하여 세 가지 표고 품종들의 담자포자에 대한 교배형 분석을 수행하였다. 그 결과 교배형의 편향된 분리비가 과거 보고와 일치하게 균주 특이적인 특성임을 확인하였다. 분석한 세 품종 중 한 품종을 제외하고 나머지 두 품종에서 편향된 분리비가 관찰된 것이다. 다음으로는 각 담자포자 유래 단핵균사들의 생장 속도와 교배형의 상관관계를 분석하였다. 그 결과 표고 단핵균사의 생장 속도는B 교배형과는 관계가 없고, A 교배형과 관계가 있음을 확인하였다. 따라서A 교배형 유전자좌에 존재하는 호메오도메인 전사인자 혹은A 교배형 유전자좌와 연관된 유전자들이 단핵균사의 생장에 영향을 줄 것으로 보인다. 버섯 신품종 육성에서 교배형의 중요성을 고려할 때, 본 연구는 효율적인 신품종 육성 전략을 세우거나 단핵균사 생장 기작을 이해하는 데 도움을 줄 것으로 기대된다. Lentinula edodes is a tetrapolar basidiomycete and its mating type is determined by two unlinked genetic loci, A and B. Theoretically, one dikaryotic strain could produce basidiospores with four different mating types in a 1:1:1:1 ratio. Previous studies have described the skewed segregation ratio of mating types among basidiospores of L. edodes. However, they were based only on morphological characteristics, such as clamp connection, to determine mating types. To clarify whether the segregation distortion of mating types is a general phenomenon in L. edodes, we analyzed the mating types of basidiospores obtained from three cultivars of L. edodes using recently developed DNA markers. We found that the skewed segregation of mating types was strain-specific, as reported previously. Among the three cultivars, one cultivar showed balanced segregation, while the other two displayed distorted segregation. We also examined the relationship between mating type and mycelial growth rate of monokaryons derived from each basidiospore. It was found that the monokaryotic mycelial growth rate was related to the A mating type but not to the B mating type. Therefore, homeodomain transcription factor genes that reside on the A locus or other genes linked to the A locus affect the growth rate of monokaryotic mycelia. Considering the importance of mating types in mushroom breeding, this study is informative for establishing an efficient breeding strategy as well as for understanding the mechanism of monokaryotic mycelial growth.

표고 A 교배형 구분을 위한 CAPS 마커의 개발 및 적용

박미정 ( Mi-jeong Park ),유림 ( Rhim Ryoo ),장영선 ( Yeongseon Jang ),가강현 ( Kang-hyeon Ka ) 한국균학회 2020 韓國菌學會誌 Vol.48 No.4

표고는 한국에서 가장 많이 소비되는 식용버섯 중 하나이다. 표고는 사극성의 교배계를 따르며, 표고의 교배형은 자웅이주성의 다른 담자균류와 마찬가지로 서로 독립적인 두 유전자좌, A와B에 의해 결정된다. 표고의A 유전자좌에는 한 쌍의 homeodomain (HD) 전사인자가 암호화되어 있으며 이들의 N말단에서 나타나는 높은 변이가A 교배형의 다양성에 중요한 것으로 알려져 있다. 본 연구에서는 표고 품종과 야생종에서 많이 발견되는 11종의A 교배형을 구분할 수 있는 CAPS 마커를 개발하고자 하였다. A 유전자좌에서 변이가 큰 부분을 PCR을 통해 증폭한 뒤 두 가지 제한효소HaeIII와EcoRI로 절단하여 DNA 단편의 크기 및 양상을 살펴봄으로써 11종의A 교배형을 서로 구별할 수 있었다. 또한 해당 방법이 이핵균주의 교배형을 확인하는 데 활용할 수 있는지도 살펴보았다. Lentinula edodes is one of the most widely consumed edible mushrooms in Korea. Mating in L. edodes is regulated by a tetrapolar system, and two unlinked genetic loci, A and B, are known to be major determinants of the mating types, as reported in other heterothallic basidiomycetes. The A locus of L. edodes encodes a pair of homeodomain (HD) transcription factors. The highly variable N-termini of these HD transcription factors contribute to the diversity among the A mating types. In this study, we developed a cleaved amplified polymorphic sequence (CAPS) marker to discriminate 11 different A mating type alleles predominant among both cultivated and wild strains. Amplification of the variable region of the A locus followed by digestion with HaeIII and EcoRI restriction enzymes enabled successful discrimination among the 11 A mating type alleles. We also evaluated the applicability of this method in the identification of two A mating types of a dikaryotic strain.

원목재배 표고버섯 물추출물의 항산화 특성

서수영 ( Sooyoung Seo ),장영선 ( Yeongseon Jang ),유림 ( Rhim Ryoo ),가강현 ( Kang-hyeon Ka ) 한국균학회 2018 韓國菌學會誌 Vol.46 No.1

We investigated the antioxidant activities of nine Lentinula edodes varieties cultivated on oak log. The total polyphenol content (TPC), total flavonoid content (TFC), and antioxidant properties of L. edodes (varieties: Dasanhyang, Chunbaegko, Poongnyunko, Soohyangko, Baekwhahyang, Chunjang 1ho, Chunjang 2ho, Sanlim 5ho, and Sanlim 7ho) were evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay and the ferric reducing antioxidant power (FRAP) assay. The TPC and TFC of mushroom extracts ranged from 5.10 to 10.88 mg gallic acid equivalent (GAE)/g and 0.76 to 5.78 mg quercetin equivalent (QE)/g, respectively. In addition, mushroom extracts exhibited DPPH radical scavenging activity of 24.22~90.41%, 2.0 mg/mL and FRAP of 0.056~0.231, 2.0 mg/mL. The Chunbaegko variety showed the highest TPC (10.88 mg GAE/g), TFC (5.78 mg QE/g), DPPH activity (38.19~90.11%, 0.2~2.0 mg/mL), and FRAP (0.075~0.231, 0.2~2.0 mg/mL).

닥나무 톱밥 첨가량에 따른 표고 생육특성 비교

정연석 ( Yeun Sug Jeong ),장영선 ( Yeongseon Jang ),유림 ( Rhim Ryoo ),가강현 ( Kang-hyeon Ka ) 한국균학회 2018 韓國菌學會誌 Vol.46 No.3

We aimed to increase the utility of Broussonetia kazinoki (paper mulberry) by using its woody parts as a substrate for Lentinula edodes cultivation. Using different mixing ratios of paper mulberry and oak tree sawdust, five types of column-type media were prepared (1.2 kg each). Two L. edodes strains (NIFoS 2462 and NIFoS 2778) were used in this study. In the first flush, numerous small mushrooms were harvested because the fruited mushrooms were densely packed on the small media. The highest productivity was obtained with a 3:1 mixture of paper mulberry and oak tree sawdust (Q. acutissima:Q. mongolica, 1:1). In particular, for NIFoS 2462, the proportion of paper mulberry in the medium positively correlated with productivity. The size of NIFoS 2778 was uniform in all media tested. By contrast, the mushroom weight of NIFoS 2462 decreased, but the other characteristics were not significantly affected. Collectively, these findings suggest that B. kazinoki sawdust could be used to grow shiitake mushrooms and that some oak sawdust substitution is also possible. Our results could increase the utility of discarded by-products, such as sawdust.

환경 스트레스 조건에 따른 화경버섯 균사의 생물발광 변화

박미정 ( Mi-jeong Park ),이효림 ( Hyorim Lee ),유림 ( Rhim Ryoo ) 한국균학회 2020 韓國菌學會誌 Vol.48 No.4

생물발광은 생물체 내에서 빛이 방출되는 현상을 말한다. 이 현상은 루시페라아제(luciferase)에 촉매되는 루시페린(luciferin)의 산화 반응에 의해 나타난다. 생물발광은 균류를 포함하여 다양한 생물 종에서 나타남이 알려져 있다. 현재까지 약 80여 종의 균류에서 생물발광이 관찰됨이 보고 되었다. 화경버섯(Omphalotus japonicus)은 한국에 자생하는 대표적인 생물발광 버섯이다. 본 연구에서는 화경버섯 균사의 생물발광에 대하여 조사하였다. 그 결과, 고체 배지에서 자란 화경버섯 균사체의 가장자리 부근에서 중심부에 비해 높은 발광이 관찰되었다. 또한 균사 표면에 상처를 내자 해당 부위에서 강한 생물발광이 관찰되었다. 이러한 생물발광의 증가는 처리 후 3시간에 최대로 나타났다. 다음으로 극단적인 온도 스트레스에 의한 생물발광의 변화에 대해서도 살펴보았다. 그 결과, 외부 자극에 의한 상처 스트레스와 달리 고온이나 저온 스트레스에 의해 균사의 생물발광이 억제됨을 확인하였다. 환경 스트레스에 의해 일어나는 생물발광 변화의 생리학적 및 생태학적 의미를 밝히기 위해 더 많은 연구가 필요할 것이다. Bioluminescence refers to the production and emission of light in living organisms. This phenomenon arises from luciferase-catalyzed oxidation reaction of luciferin. Bioluminescence is widely observed in marine vertebrates and invertebrates, as well as in some microorganisms and fungi. To date, approximately 80 species of fungi have been reported to be luminous. One such example is Omphalotus japonicus, which is a luminous fungus found in Korea. In this study, we examined the bioluminescence of Omphalotus japonicus mycelia. Light emission was detected at the edges of mycelia grown on solid agar medium. Notably, the intensity of bioluminescence was found to be significantly enhanced following wound induction. The increase in light intensity peaked at 3 h after mechanical damage. We also investigated the effects of extreme temperatures on bioluminescence. Unlike mechanical damage, high and low temperatures repressed the light emission from mycelia. Further investigations are required to reveal the physiological and ecological properties of fungal bioluminescent responses to environmental stresses.

분리형 용기를 이용한 표고버섯 톱밥재배 연구

정연석 ( Yeun Sug Jeong ),장영선 ( Yeongseon Jang ),유림 ( Rhim Ryoo ),가강현 ( Kang-hyeon Ka ) 한국균학회 2019 韓國菌學會誌 Vol.47 No.4

Currently, plastic bags are being used for sawdust cultivation of shiitake mushroom. However, due to serious environmental problems caused by the use of plastic bags, we studied the efficacy of bottle cultivation method to replace the sawdust bag method. Small detachable plastic bottles (400 g capacity) filled with Quercus spp. sawdust and wheat bran (4:1 w/w) media were incubated for 80 and 120 days. The weight loss (%) of the media was higher for the NIFoS 2464 strain at an approximate light intensity of 300 Lux than light intensity of 500 Lux; the light intensity was associated with the loss of sawdust medium-weight during the cultivation period. The highest yield was observed when the strain was cultivated for 80 days under dark conditions, 40 days under 500 Lux light, and air circulation fan speed of 30 rpm. When incubated for 120 days, mushroom yield in the bottle media was higher at 40 days of light exposure than 20 days of light exposure. In the bottle media incubated for 80 days under dark conditions, the mushrooms fruited due to repetitive water spraying on the top of the media and light stimulation during the fruiting period. The media could be separated from the bottles because the media shrank after the first harvest. These separated plastic bottles could be re-used for mushroom cultivation, thereby reducing the amount of plastic waste.

다양한 기질에 대한 표고버섯 톱밥재배 가능성

정연석 ( Yeun Sug Jeong ),장영선 ( Yeongseon Jang ),유림 ( Rhim Ryoo ),가강현 ( Kang-hyeon Ka ) 한국균학회 2019 韓國菌學會誌 Vol.47 No.4

We investigated the effect of different media components on the characteristics and productivity of Lentinula edodes cultivated in sawdust. Ten substrates were used for this study. Soybean and cotton seed meal were unsuitable substrates as fungal mycelia did not grow to maturity during the incubation period. Lentinula edodes (NIFoS culture number 2462) was grown on the ten substrates, and soybean hull, mixed medium, cotton seed hull and corn grain showed greater effect on the productivity than wheat bran. However, wheat bran induced higher productivity in NIFoS 2778 than the other substrates. Sawdust medium with soybean meal produced smaller fruiting bodies than the other substrates. In contrast, corn meal media produced larger mushrooms than the other substrates. The external characteristics of the mushrooms varied based on the substrates upon which they were grown. This is not surprising given that the substrates differed with respect to carbohydrate and protein content, e.g., cotton seed and soybean meal contained a higher crude protein and crude ash than the other substrates tested.

신품종 표고버섯 ‘산장향’의 교배 육성 및 재배 특성

박영애 ( Youngae Park ),장영선 ( Yeongseon Jang ),유림 ( Rhim Ryoo ),이봉훈 ( Bonghun Lee ),가강현 ( Kang-hyeon Ka ) 한국균학회 2019 韓國菌學會誌 Vol.47 No.2

본 실험은 ‘산백향’과 ‘장안1호’의 단포자 간 교잡에 의해 새로운 품종을 만들었으며 그 이름을 ‘산장향’이라 명명하였다. ‘산장향’의 주요 형태적 특성은 갓이 평반구형이며, 갓의 직경은 67.1 mm, 갓 두께는 16.9 mm이었다. 주름살 측면 모양은 부정형이며 주름의 밀도는 조밀하고 주름살의 폭은 좁다. 인편은 흰색 또는 옅은 갈색으로 갓 전체에 분포하였다. 대조품종인 ‘산백향’보다 대의 길이가 길고, 갓 색깔은 갈색계열이나 적색이 진하였다. 대의 표면에는 털이 있고 대와 같은 크림색이었다. 버섯은 산발 발생으로 솎아주기 작업을 적게 할 수 있었다. 자실체의 발이온도는 평균 15-19℃으로 중온성이었다. ‘산장향’의 발생시기는 봄, 가을 이었다. 자실체 발생은 3차까지 진행하였고, 1차(89%), 2차(4%) 및 3차(7%)의발생비율을보였다. A new cultivar ‘Sanjanghyang’ was bred from monokaryotic strains of ‘Sanbaekhyang’ and ‘Jangan 1ho’. Pileus was flat, round, and reddish brown. The diameters of the pilei and stipe length of the fruiting bodies were 67.1 mm and 16.9 mm, respectively. The scales were white or slightly brown and distributed evenly. The gill density was sparse and showed a rippled texture. The stipe was cream in color and the fluff was medium. ‘Sanjanghyang’ had a short cultivation period and fruiting bodies occurred sporadically. Temperature for fruiting body formation was a medium, between 15 to 19°C. ‘Sanjanghyang’ was different from ‘Sanbaekhyang’ with regard to its pileus diameter (67.1 mm) and autumn and spring fruiting body production period. ‘Sanbaekhyang’ had pileus diameter of 74.7 mm, and fruiting body formation occurred in spring and autumn. The rate of fruiting body formation was 89% (first flush), 4% (second flush), and 7% (third flush).