• Journal of Mushroom
• /
• v.19 no.2
• /
• pp.88-95
• /
• 2021

This study was carried out to breed new variety of Pleurotus nebrodensis. We have collected and tested characteristics of genetic resources from domestic and abroad since 2015. The varieties of P. nebrodensis from China are grown by farmers, but those have been unstable fruiting and are weak against bacterial diseases. To solve this problem, we bred the unique domestic variety 'Uram' of P. nebrodensis and the results of the characteristic test for the new 'Uram' are as follows. The proper temperature for the mycelial growth was 26~29℃ and fruit body growth temperature was 15~18℃. It was similar to the control variety KME65035 of P. nebrodensis in the pileus form of a flat and white color. The number of days required for initial fruting was 5 days for bottle cultivation and 6 days for bag cultivation which was 2-4 days shorter than that of the control variety. The pileus diameter was 32.6-37.0 mm which was smaller but the fruit body length was 130.4 mm, which was longer than those of the control variety. The effective number of fruit bodies was 1.8 in bottle cultivation and 2.9 in bag cultivation, which was more than those of the control variety. The yield rate was 93.3-100%, which was more stable than those of the control variety. In bottle cultivation and bag cultivation, the yield was 173.1 g/bottle (1100 cc) and 283.4 g/bag (1.2 kg), respectively, which was 25-44% higher than those of the control variety 138.0 g/bottle (1100 cc) and 197.4 g bag (1.2 kg). When incubating the parent and control varieties of 'Uram', the replacement line was clear and as a result of mycelial DNA RAPD-PCR reaction, the band pattern was different from that of the parent and control varieties, confirming the hybrid species.

• 한국균학회소식:학술대회논문집
• /
• 2015.05a
• /
• pp.53-53
• /
• 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.