• Biotechnology and Bioprocess Engineering:BBE
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• 제9권6호
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• pp.440-446
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• 2004

Interest in in vitro study of entomopathogenic fungi, including Cordyceps species, has been increasing due to their valuable bioactive compounds and biocontrol effects. Among Cordyceps species, in vitro stromata of C militaris has been successfully produced and cultivated for industrial purposes. However, genetic study on in vitro stromata formation of C militaris has not been carried out yet. Here, relationship between mating system and perithecial stromata formation of C militaris is reported. Mating system was determined by observing perithecial stromata formation from mono-ascospore cultures and their pair-wise combinations. Certain combinations of mono-ascospore strains produced perithecial club-shaped stromata, whereas other combinations produced either no stromata or only abnormal non-perithecial stromata. Similarly, mono­ascospore cultures without combination produced either no stromata or only abnormal non­perithecial stromata. Despite obvious heterothallism, self-fertility was occasionally observed in few strains of C militaris. These observations indicated that C militaris behaves as a bipolar het­erothallic fungus and requires two mating compatible strains in order to produce regular club­shaped perithecial stromata, a fundamental requirement for its industrial cultivation.

• 한국버섯학회지
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• 제13권1호
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• pp.41-49
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• 2015

표고(Lentinula edodes)는 우리나라의 주요 임산버섯으로 최근 표고 톱밥재배가 정착됨에 따라 급속도로 확산되고 있으나 국내 환경에 적합하고 재배시기를 다변화 할 수 있는 균주의 육성이 시급하다. 따라서 9개의 균주를 시험재배하여 생산성이 높고 자실체 특성이 우수한 4개 균주(SANJO701HO, FMRI2534, FMRI2337, FMRI2613)를 모균주로 선발하였고 균주당 각각 20개씩 총 80개의 단핵균주를 분리하여 117개의 교배균주를 육성하였다. 그중 산조701호와 FMRI2337의 교배조합이 다른 조합의 교배균주들보다 평균생장속도가 10% 가량 낮았으며 이들 교배균주의 배양 및 생리적 특성을 비교 조사하였다. 따라서 본 연구는 우수한 톱밥재배용 품종개발의 기반조성을 마련하고자 하였으며, 차후 교배균주에 대한 생산성 검정의 연구가 필요하다.

• 한국균학회지
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• 제51권4호
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• pp.389-396
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• 2023

본 연구에서는 교배형 분석을 활용하여 표고의 교배균주를 육성하고 그로부터 발생한 자실체의 특성을 알아보고자 하였다. 단핵균주 분리에는 산백향과 청흥1호 품종을 이용하였는데, 산백향은 버섯으로부터 100여개의 단포자를 분리하였으며, 청흥1호는 원형질체 분리 및 재생과정을 통해 단핵균주를 얻었다. 이렇게 얻는 단핵균주는 우선 교배형 분석을 실시하였으며, 교배 가능한 조합을 확인하고 총 60개의 균주를 교배하였다. 톱밥재배는 2 kg 사각블럭배지를 이용하였으며, 배양 120일 후에 버섯 발생처리를 하였다. 그 결과 60균주의 교배성공률은 100%에 이르렀으며, 그 중 55개인 91.7%의 균주에서 버섯이 발생하였다. 모든 균주에서 정상적인 버섯이 발생하였으며, 그 중 10개 균주에서는 주름살을 온전하게 형성하지 않는 기형버섯도 발생하는 것을 확인하였다. 버섯의 형태적인 특성은 매우 다양하게 나타났으며, 이러한 특성을 사전에 파악하기 위해서는 교배모본의 특성을 평가할 수 있는 연구가 더 필요하다.

• 현장농수산연구지
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• 제20권1호
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• pp.49-54
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• 2018

느티만가닥버섯 해송이 품종의 우수 균주를 선발하기 위하여 각기 다른 자실체에서 분리한 균을 Di-mono 교배하여 균을 선발하였다. 교배한 균주를 PDA에서의 균사생장 조사를 통하여 균사생장이 좋은 3균주를 선발하였다. PDA배지에서 10일간 배양했을 때 균사생장은 3번 균주가 62mm로 가장 빠르고, 7번 균주가 58mm, 그리고 2번, 10번, 9번순이었다. 선발된 3균주, 즉, KNCAF-H-3, KNCAF-H-7, KNCAF-H-2는 톱밥배지에서 10일간 배양한 결과 각각 79mm, 76mm, 73mm의 균사생장을 보여, 대조 품종인 그린피스 5호의 55mm보다 균사생장이 좋았다.

• 한국버섯학회지
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• 제9권3호
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• pp.96-100
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• 2011

다양한 느타리버섯 품종간의 교잡을 하였으며 균상재배용으로서 유용한 특성을 갖는 느타리버섯 품종을 육종코자하였다. 그 중 화성1호에서 단핵균을 분리한 후 수한1호 이핵균과 di-mono교배법으로 육성된 MST247ns가, 세균성 갈변병에 약하고 저온기 재배시 어려움이 많은 수한1호의 단점을 보완하면서 우수한 특성을 가지므로 최종 선발하여 "화성2호"로 명명하였다. 수한1호와 비교하여 대가 길고 조직이 무르지 않고 단단하였다. 화성2호는 중고온성인 수한1호와 비교하여 낮은 온도에서도 갓이 작고 대가 길며 저장성이 좋은 특성을 더욱 잘 나타내었다. 이러한 특성은 시장에서 선호하는 버섯으로서, 낮은 온도에서의 재배관리가 비교적 용이한 가을철에서 이른봄철까지 농가에서의 난방비 부담을 줄여주며 양질의 버섯을 생산할 수 있어 농가의 소득증대에 많은 도움을 줄 것으로 기대된다. '화성2호'의 주요특성으로는 가. 적정 균사배양온도는 $25-30^{\circ}C$이다. 나. 버섯발생 온도는 $8-15^{\circ}C$, 자실체의 적정 생육온도는 $9-14^{\circ}C$으로 가을철부터 봄철까지의 기간에 재배하기에 좋은 품종이다. 다. 수한1호와 비교하여 대가 길고, 갓은 얕은 깔때기형이다. 라. 낮은 온도에서 재배시에도 대가 길고 곧으며 대의 표면이 백색이며 조직이 단단하면서 탄력이 있으므로 저온기의 생육관리가 비교적 쉽고 저장성이 우수하다.

• The Plant Pathology Journal
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• 제31권2호
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• pp.97-107
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• 2015

Alteration of genetic make-up of the isolates and mono-sporidial strains of Tilletia indica causing Karnal bunt (KB) disease in wheat was analyzed using DNA markers and SDS-PAGE. The generation of new variation with different growth characteristics is not a generalized feature and is not only dependant on the original genetic make up of the base isolate/monosporidial strains but also on interaction with host. Host determinant(s) plays a significant role in the generation of variability and the effect is much pronounced in monosporidial strains with narrow genetic base as compared to broad genetic base. The most plausible explanation of genetic variation in presence of host determinant(s) are the recombination of genetic material from two different mycelial/sporidia through sexual mating as well as through parasexual means. The morphological and development dependent variability further suggests that the variation in T. indica strains predominantly derived through the genetic rearrangements.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 춘계학술대회 및 임시총회
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• pp.53-53
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• 2015

Agrobacterium tumefaciens-mediated transformation(ATMT) of Flammulina velutipes was used to produce a diverse number of transformants to discover the functions of gene that is vital for its variation color, spore pattern and cellulolytic activity. Futhermore, the transformant pool will be used as a good genetic resource for studying gene functions. Agrobacterium-mediated transformation was conducted in order to generate intentional mutants of F. velutipes strain KACC42777. Then Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into F. velutipes. This method is use to determine the functional gene of F. velutipes. Inverse PCR was used to insert T-DNA into the tagged chromosomal DNA segments and conducting sequence analysis of the F. velutipes. But this experiment had trouble in diverse morphological mutants because of dikaryotic nature of mushroom. It needed to make monokaryotic fruiting varients which introduced genes of compatible mating types. In this study, next generation sequencing data was generated from 28 strains of Flammulina velutipes with different phenotypes using Illumina Hiseq platform. Filtered short reads were initially aligned to the reference genome (KACC42780) to construct a SNP matrix. And then we built a phylogenetic tree based on the validated SNPs. The inferred tree represented that white- and brown- fruitbody forming strains were generally separated although three brown strains, 4103, 4028, and 4195, were grouped with white ones. This topological relationship was consistently reappeared even when we used randomly selected SNPs. Group I containing 4062, 4148, and 4195 strains and group II containing 4188, 4190, and 4194 strains formed early-divergent lineages with robust nodal supports, suggesting that they are independent groups from the members in main clades. To elucidate the distinction between white-fruitbody forming strains isolated from Korea and Japan, phylogenetic analysis was performed using their SNP data with group I members as outgroup. However, no significant genetic variation was noticed in this study. A total of 28 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4210 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr01, chr04, chr07,chr11 regions were identified to be associated with white fruitbody forming. White and Brown Fruitbody strains can be used as an identification marker for F. veluipes. We can develop some molecular markers to identify colored strains and discriminate national white varieties against Japanese ones.

• 한국수정란이식학회지
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• 제26권1호
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• pp.79-84
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• 2011

This study was performed to identify the differentially methylated region (DMR) and to examine the mRNA expression of the imprinted H19 gene in day 35 of SCNT pig fetuses. The fetus and placenta at day 35 of gestation fetuses after natural mating (Control) or of cloned pig by somatic cell nuclear transfer (SCNT) were isolated from a uterus. To investigate the mRNA expression and methylation patterns of H19 gene, tissues from fetal liver and placenta including endometrial and extraembryonic tissues were collected. The mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. Bisulfite analyses demonstrated that the differentially methylated region (DMR) was located between -1694 bp to -1338 bp upstream from translation start site of the H19 gene. H19 DMR (-1694 bp to -1338 bp) exhibits a normal mono allelic methylation pattern, and heavily methylated in sperm, but not in oocyte. In contrast to these finding, the analysis of the endometrium and/or extraembryonic tissues from SCNT embryos revealed a complex methylation pattern. The DNA methylation status of DMR Region In porcine H19 gene upstream was hypo methylated in SCNT tissues but hypermethylated in control tissues. Furthermore, the mRNA expression of H19 gene in liver, endometrium, and extraembryonic tissues was significantly higher in SCNT than those of control (p<0.05). These results suggest that the aberrant mRNA expression and the abnormal methylation pattern of imprinted H19 gene might be closely related to the inadequate fetal development of a cloned fetus, contributing to the low efficiency of genomic reprogramming.