NGS data was yielded by using Illumina Hiseq platform. The short reads were filtered by quality score and read length were mapped against the reference genome (KACC42780). Genome-wide reanalyzed data of Flammulina strains were compared against the reference genome to establish a genome-wide single nucleotide polymorphism (SNP). The rate of mapping differences between the strains reflected in the strain variation in its result. The genome-wide SNPs distribution divided into types of homozygous SNP and heterozygous SNP moreover all of the strains demonstrated a wide variation in all of the regions. In the further study of topological relationship between the collected strains, phylogenetic tree was separated into 3 major groups. Group I contained F. velutipes var. related strains of ASI 4062, 4148, 4195. Group Ⅱ contained strains that were different species of ASI 4188 F. elastica, ASI 4190 F. fennae, and ASI 4194 F. rossica. The other 19 strains F. velutipes were classified as a single group. Polymorphic SNPs of F. velutipes strains representing the phylogenetic segregation of whiteand brown-fruiting body forming groups were compared. As previously reported, white gene expression is recessive to brown in fruiting body color gene expression. The white strains produced 131,874 SNPs to be aa type and homozygous from of SNP. 407,947 SNPs were detected as AA, Aa type from the brown-fruiting body of SNP. We constructed a SNP matrix with 8 white strains and 12 brown strains. To develop the molecular marker related in to fruiting body color and geographical origin, we isolated 240 SNPs from the white-and brown-fruiting body forming. To determine the chromosome relationship on polymorphic SNP between Korea and Japan strains producing white-fruiting body, we analyzed that the Korea white strains detected 185,695 SNPs and the Japan white strains produced 263,811 SNPs. Using the constructed SNP matrix with 3 Korea white strains and 3 Japan white strains, the experiment generated 475 SNPs of phylogenetic SNPs fromKorea and Japan white-fruiting body. As a result, we regarded as they are potentially related to the white color. White and brown color and origin specific SNPs could be used as an identification marker for selection of F. veluipes strains in the breeding program.

Many basidiomycetes, especially mushroom species, thrive on saccharides that they obtain from the decomposition of cellulose, hemi-cellulose and or lignin. Cellulose is degraded by specific enzymes called cellulases. Cellulases play an important role in the biosphere by recycling cellulose, the most abundant carbohydrate produced by plants. Also cellulases have many potential biotechnologic and industrial applications. Detection of cellulose degrading activity is commonly performed on carboxymethylcellulose CMC medium in combination with a stain that reveals the degraded CMC. In order to efficiently screen the F. velutipes knockout library, obtained through Agrobacterium-mediated transformation (ATMT), for those important enzymes we compared two different staining methods i.e. Congo Red and Gram’s Iodine (as reported in R. C. Kasana et al 2008). The latter stain showed strongly enhanced detection in time and intensity, facilitating mass screening of our mutant database. Using Gram’s iodine and square petri dishes containing up to 9 colonies at a time we can now rapidly screen multiple mutants in a short period. We are going to find the genes. Inactivated genes of mutants with altered cellulase degradation activity will be identified and cloned for further analysis.