Di-Mon 교잡법에 의한 큰느타리버섯 교잡주의 특성

신평균 외 한국버섯학회 2004 한국버섯학회지 Vol.2 No.2

큰느타리버섯은 식용 및 약용으로 쓰이는 부가가치가 높은 버섯으로서 한국 고유의 육성품종 개발이 절실하다. 이에 새로운 큰느타리버섯 품종을 육성하고자 시중에 고가로 판매되고 있는 ASI 2547균주의 단포자를 분리하고 22개의 수집균주와 Di-Mon 교배를 실시하여 꺽쇠연결체를 가진 253균주중에서 자실체를 형성하는 19균주를 선발하여 그 중에서 형태적으로 다양한 7균주를 선택하여 RAPD로 분석한 결과 Hybrid H6 균주가 형태학적으로는 양친과 유사하나 유전자 증폭산물은 재조합된 교배체임이 확인되었다. 따라서 Di-Mon 교배법이 분자생물학적 동정과 결합됨으로서 손쉽고 빠른 교배법으로 새로운 균주육성에 사용할 수 있으리라 사료된다. Pleurotus eryngii is not edible and medicinal mushrooms indigenous to Korea. To improvement of strain suitable to the geographic setting of Korea, we are mated with 22 dikaryons and 47 monokaryons isolated from Pleurotus eryngii ASI 2547 by Di-Mon mating. 19 strains forming fruit body obtained from clamped 253 bred strains. 7 excellent strains are selected from 19 bred strains by various morphological features of fruit body. Among the selected 7 strains, H6 strain were identified into ASI 2547-like recombinant hybrids with URP uniprimer by RAPD analysis. This suggested that Di-Mon crossing is one of rapid and easy breeding method for strain improvement with molecular techniques.

ITS 부위의PCR-RFLP 및 STS 마커를 이용한 차가버섯의 종 및 계통간 유연관계 분석

신평균 외 한국버섯학회 2009 한국버섯학회지 Vol.7 No.4

최근들어 수입량이 급증되고 있는 차가버섯의 분류체계를 확립하고, 이들 종 및 계통간의 유연관계 확립과 품종 구분을 위해 계통학적 정보를 지닌 ITS 영역의 염기서열, PCRRFLP 및 STS 프라이머를 사용하여 종 특이적인 마커를 개발하였다. 시베리아 캄차카 반도에서 수집된 74008 균주의 염기서열을 이용하여 Inonotus spp.의 유연관계를 분석한 결과 2개의 그룹으로 나뉘어 졌고, I. obliquus DSM 856P균주와 약 98%의 가장 높은 유사성을 나타내어 차가버섯임이 확인되었다. 또한 ITS 증폭산물을 제한효소로 처리하여 밴드 패턴을 비교하였을 때 종 및 계통에 따라 밴드가 다르게 나타났으며, STS primer를 이용하여 증폭산물을 비교하였을 때 종간에는 밴드패턴이 다르나 계통내에서는 동일한 밴드패턴을 보였다. 따라서 차가버섯의 품종 구분을 위해서는 STS 마커와 PCR-RFLP를 동시에 사용함으로서 품종 구분이 좀 더 명확하리라 사료된다. Because the import of Inonotus obliquus have been rapidly increased in Korea, we developed the Inonotus species-specific marker by using various sequences including ITS sequences, PCR-RFLP and STS primers and used this marker to determine both genetic relatedness and strains discrimination of Inonotus spp. Total 17 different Inonotus spp. were examined by using ITS sequences and classified into 2 different groups. One strain, ASI74008 isolated from Kamchaka island of Siberia, showed the high sequence identity (98%) at the nucleotide level to the other I. obliquus DSM strain, indicating the ASI74008 belong to I. obliquus species. Comparison of banding patterns after restriction enzyme digestions with PCR amplicons of ITS region revealed some variations depending on the species and strains. However, PCR products amplified with STS primer showed species specific patterns.Therefore, use of both STS primers and PCR-RFLP could help for better strain identification.

무포자느타리 선발을 위한 단핵화 균주의 분리 및 Spo11 마커의 이용

신평균 외 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4

무포자형성 자연돌연변이 균주의 선발 및 특성검정 결과 ASI 2069 균주가 무포자이면서 수량성이 높으나 상품적 가치가 없어 원형질체 재생에 의한 단핵화로 Nh36 (neohaplont 36) 등 4균주를 분리하였다. 원형1호(ASI 2180) 및 무포자느타리간의 단핵교배 (Mon-Mon)를 시도한 결과 128교배조합수를 얻어 이 중 30균주를 특성 검정한 다음 자실체형성 및 포자비산량 조사에 의해 소포자형성 유망 13균주를 선발하였다. 무포자균주의 스크리닝법을 개발하기위해 포자형성 및 감수분열에 관여하는 helicase, DMC1(recombinase) 및 Spo11(topoisomerase II)유전자를 이용하여 PCR 증폭한 결과 단핵화균주 Nh36 및 교배체 2균주 중에서 Spo11유전자가 증폭되지않아 무포자 균주 스크리닝 방법으로 가능하리라 사료된다. For the development of a sporeless strain of P. ostreatus we used sporeless strain ASI 2069. We have recovered both nuclear types of strain ASI 2069 as monokaryons of nh9, nh15, nh26 and nh36 (here after referred to as neohaplonts) by protoplasting the mycelium. Crosses between neohaplonts and SSI's(single spore isolates) obtained from a sporulating commercial strain ASI 2180. Five excellent strains are selected from 30 bred strains by quality of fruitbodies and spore number. To development of molecular markers linked to sporeless strain of P. ostreatus, we are screened helicase, recombinase(DMC1) and topoisomerase(Spo11) genes related meiosis by PCR and sequencing. Among three genes, Spo11 gene was identified into molecular marker of sporeless from neohaplonts and bred strains of P. ostreatus.

신령버섯의 기형으로부터 배양불능세균의 PCR 검정

신평균 외 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4

신령버섯은 항암작용과 더불어 면역강화작용 등 약용으로서 이용되고 있습니다. 최근에는 신령버섯 재배농가에서 새로운 병징 현상이 보고되어 내생세균에 대한 분자생물학적 방법으로 조사하였다. 병징을 나타내는 수집된 기형버섯을 eubacterial 16S rDNA 영역을 이용하여 PCR한 결과 기형버섯만 증폭되었다. 이 영역을 부분적으로 염기서열을 결정한 결과 CFB bacterium과 유사성이 가장 높았고, 이 염기서열을 이용하여 프라이머를 디자인한 후 nested PCR를 부위별로 확인한 결과 기형을 일으킨 갓의 주름살 부위에서 가장 강하게 증폭되었고 포자수확도 되지 않았으며 배양불능세균 group인 CFB bacterium임을 확인하였다. Agaricus blazei Murill is a important medicinal mushroom for a powerful immune system builder and tonic. Currently, it is known about a new disease phenomenon that appears to be occurring on a number of mushroom farms. We described a straightforward approach in which molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria infected with the Agaricus blazei. The 16S rDNA was amplified with universal eubacterial primers directly from pure cultures of Agaricus blazei mycelium and fruit body. The 16S rDNA sequences were almost identical (96 to 97% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belong to the uncultural bacterium phylogroup. PCR detection of uncultural bacterium in the malformed tissues of Agaricus blazei were carried out by using 16S rRNA sequenced specific probe. It was strongly amplified at the malformed pileus region of fruit body and also spore print was impossible.

잎들깨 수량과 단백질체 발현에 미치는 UV-B의 영향

홍성창,황선웅,장안철,신평균,장병춘,이철원 한국환경농학회 2006 한국환경농학회지 Vol.25 No.1

Characterization of a Ca2+ / Calmodulin Dependent Phosphorylated Protein Expressed at the Early Stages of Soybean Germination

Pyung Gyun Shin,Gun Do Kim,Yong Hwa Cheong,Sang Yeol Lee,Jeong Dong Bahk,Jong Chan Hong,Moo Je Cho ) 생화학분자생물학회 1993 BMB Reports Vol.26 No.2

Using [γ-^(32)P] labelled ATP, an endogenous phosphorylated protein whose molecular weight was 52 kDa has been identified and characterized. It was rapidly appeared during the early stage of soybean germination. It was distinctively phosphorylated at the very early stages of cell growth and differentiation and slowly declined after 24 h. Therefore, the phosphorylated 52 kDa protein can be postulated to play significant roles in the control of embryonic cell growth and development. The 52 kDa phosphorylated protein was identified to be mainly located in the cytosolic fraction of soybean cell. Phosphorylation of this protein was dependent on Ca^(2+) and calmodulin but not on phospholipids. The requirement of free Ca^(2+) to phosphorylate this protein can be supplemented enough by soybean cell itself without the additions of Ca^(2+) from the external sources. The phospho-amino acids were identified as serine and threonine residues, which represented that the 52 kDa protein contained at least two phosphorylation sites.

New Ferula mushrooms of inter-specific Pleurotus ferula x Pleurotus tuoliensis using cytoplasmic hybrid with microsatellite marker

Pyung Gyun Shin,Min-Ji Oh,Yeun-Lee Oh,Kab Yeul Jang,Won Sik Kong,Young Bok Yoo 한국버섯학회 2017 버섯 Vol.21 No.2

The Pleurotus tuoliensis is a mushroom named Chinese ‘Bai-Ling-Gu’ from species of Pleurotus nebrodensis. we are mated for selection of cytoplasmic hybrid by mitochondria microsatellite marker and the method of Mon-Mon mating between monokaryotic strains derived from Pleurotus ferula ASI 0629 (Beesan No.1) and Pleurotus tuoliensis ASI 0663 (Baekryung 20). The cytoplasmic hybrids were identified 8 strains contained nuclear DNA bands of 'Baekryung 20' and mitochondrial DNA of monokaryon strains derived from Pleurotus ferula ASI 0629 (beesan No.1). The KiMB-Plft-15-81 was shown the best cultural characteristics, selected to be a new cultivar and designed as 'Baekwhang'. The 'Baekwhang' cab be grown without an extra after-ripening period, can utilize bottle cultivation of Pleurotus eryngii. And also the ‘Seolwon’ is similar to the existing Pleurotus tuoliensis in shape and physiological characteristics, formed high stipe. We therefore expect that this new strains will substitutional goods of Pleurotus eryngii.

Functional characterictics and breeding of new commmercial strains Pleurotus ferula using Di-Mon mating and physiological assay

Pyung Gyun Shin,Min-Ji Oh,Yeun-Lee Oh,Kab Yeul Jang,Won Sik Kong,Young Bok Yoo,Hyung-Jun Noh,Keun-Ki Kim,Gun-Do Kim 한국버섯학회 2017 버섯 Vol.21 No.2

There are three kinds of Pleurotus eryngii : P. eryngii var. eryngii (P. eryngii), P. eryngii var. ferula (P. ferula : A-Wei-Mo) P. eryngii var. nebrodensis (P. nebrodensis : Bai-Ling-Gu). We bred P. ferula using Di-Mon mating and physiological assay and selected as follow (1) ‘Beesan No.1’ produced high yields, (2) ‘Beesan No.2’ was excellent morphological shape and anti-adipogenic Activit, (3) ‘Ergo’ included high ergothioneine such as anto-oxidant material, (4) ‘GW10-45’ included highest ergothioneine such as anto-oxidant material, (5) ‘Maeksong’ included high ergothioneine and effect of anti-inflammatory. We suggest that there are able to food-medicine materials.

Identification of Multiple Active Forms in Cellulase-xylanase of Aspergillus sp . 8-17 by Active Staining

( PYUNG GYUN SHIN,JUN BAE AHN,CHANG YOUNG KIM,WON HWA JEONG,JIN CHANG RYU ) 한국미생물생명공학회 1998 Journal of microbiology and biotechnology Vol.8 No.1

Role of Siderophore in Biological Control of Plant Pathogens and Increased - Growth Response of Cucumber by Fluorescent Pseudomonas Pf3

Pyung Gyun Shin,Min Chel Kim,Hee Kyu Kim,Chang Seuk Park,Moo Je Cho 한국미생물생명공학회(구 한국산업미생물학회) 1993 한국미생물생명공학회 학술발표초록집 Vol.1993 No.1