• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.6
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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.

• Journal of Mushroom
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• v.13 no.1
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• pp.41-49
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• 2015

In order to breed new strains of oak mushroom, Lentinula edodes, on the sawdust cultivation, we collected 10 from Korea, Taiwan and China respectively and examined somatic incompatibility, morphological features of fruiting body and productivity. Four strains(SANJO701HO, FMRI2534, FMRI2337 and FMRI2613) shown remarkable results, were confirmed with parental strains. 80 monokaryotic strains derived from the selected 4 parental strains, were selected and 117 hybrid strains were made by mono-mono mating. Aslo, Physiological characteristics were investigated. Average of the mycelial growth of hybrid strains of mating combination SANJO701HO-FMRI2337 was approximately 10% lower than other mating combinations. Overall, This study was founded to develop new varieties of L. edodes. The productivity of new 117 strains on sawdust cultivation needs continued research.

• The Korean Journal of Mycology
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• v.51 no.4
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• pp.389-396
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• 2023

In this study, dikaryotic strains of Lentinula edodes were generated by mono-mono hybridization using mating type analysis and their fruiting body characteristics were investigated. Approximately 100 monokaryotic strains were isolated from the basidiospores of Sanbaekhyhang, and homokaryotic strains were isolated from Chungheung 1ho using protoplast isolation and regeneration. Their mating types were evaluated and a total of 60 dikaryotic strains were hybridized. Using these strains, fruiting bodies were produced and their characteristics were examined after cultivation on sawdust media. The results indicated that the rate of hybridization was 100% and that 55 of 60 strains formed fruiting bodies. These showed normal pileus and gill structures; however,10 strains also produced fruiting bodies with abnormal pileus and gill structures. The weight and size of the fruiting bodies differed depending on the strains. Overall, further studies are needed for predicting the characteristics of hybridized strains based on their parental strains.

• Journal of Practical Agriculture & Fisheries Research
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• v.20 no.1
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• pp.49-54
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• 2018

To select the superior strain of Haesongi, Hypsizigus marmoreus, strains were isolated by Di-Mono mating with isolates from different fruit bodies. Three strains were selected to superior isolates that are good mycelial growth on PDA medium for 10days. When cultured on PDA medium for 10 days, strain No. 3 and strain No. 7 showed mycelial growth of 62mm and 58mm, respectively. Mycelial growth was good in the order of strain No. 2, 10, and 9. The three selected strains, KNCAF-H-3, KNCAF-H-7 and KNCAF-H-2, were cultured in sawdust medium for 10 days and showed mycelial growth of 79mm, 76mm and 73mm respectively. The mycelial growth of the selected three cultivars was better than that of Greenpeace No 5, a control cultivar grown at 55mm.

• Journal of Mushroom
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• v.9 no.3
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• pp.96-100
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• 2011

MST247ns(Hwaseong #2) was developed by the method of Di-mon mating between monokaryotic strains derived from "Hwaseong #1" and dikaryotic strain "Suhan #1". The optimum temperature of mycelial growth was $25-30^{\circ}C$. The optimum temperature of primordia formation and fruiting body development were $8-15^{\circ}C$ and $9-14^{\circ}C$. Days of primordia formation were 4-5 days later Suhan #1. The stipes were longer than "Suhan #1". The surfaces of stipe were white and the tissues got harder and more elastic. Therefore, the management of growth environment under low temperatures was relatively easy and storability got much better.

• The Plant Pathology Journal
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• v.31 no.2
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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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• 2015.05a
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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.

• Journal of Embryo Transfer
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• v.26 no.1
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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.