Flammulina velutipes is an edible basidiomycete with increasing agricultural and biotechnological importance. However, genetic manipulation and breeding of this organism are restricted because of the lack of knowledge about its genomic information. The aim of this project is whole genome sequencing of Korean F. velutipes strain and finding molecular markers based on its sequence information. In genomic resources, we selected a wild indigenous strain and isolated monokaryons by spore dilution. An electrophoretic karyotype of Korean F. velutipes was obtained using contour-clamped homogeneous electric field ( CHEF ) gel electrophoresis. The monokaryotic progenies (4019-20 and 4019-18) showing full compatibilty and a dikaryotic strain (4019-20 × 18) had 6-8 chromosomes, with sizes ranging from 1.6 to 5. 8 Mb. Among the 3 strains that were examined, strain 4019-20 resolved into at least 7 chromosomes, with a total genome size of approximately 26.7 Mb. We constructed two high-quality BAC libraries, fingerprinted 30 × genome equivalent BAC clones and generated BAC end sequences from 7 680 BAC clones by robust and cost effective protocols. The physical map has been constructed consisting of 11 major contigs sizing 35 284 CB Units in total. GS-FLX titanium system was applied to produce 30 X genome equivalent draft sequence of the genome. Approximately 17 Mb of sequences produced by GS-FLX titanium system was anchored onto the physical map. We also produced 19. 3 Mb of draft sequences from the selected BAC clones mapped by using both of sequencing technologies of Sanger and NGS. After that, another monokaryon 4019-18 compatible to 4019-20 and dikaryon 4019-2018 made by crossing between these two monokaryons were resequenced by GAIIx for comparing against the map of 4019-20. When comparing these two genome, we can find large numbers of SNPs and INDELs. Probably these data can be used for effective tools to protect strains and to select in the breeding system.
Flammulina velutipes Genome Sequencing Re-sequencing SNP ( Single Nucleotide Polymorphism) BAC Library Monokaryon Immunomodulation
Won-Sik Kong Jong-Guk Kim Arend F. van Peer Kab-Yeul Jang Pyung-Gyun Shin Young-Bok Yoo Young-Jin Park Byoung-Moo Lee Hyungtae Kim Chang-Soo Lee
National Institute of Horticultural and Herbal Science, RDA Department of Life Science and Biotecnology, Kyungpook National University, Korea National Academy of Agricultural Science,RDA Domestic Genomics Division, Macrogen Inc. Department of Applied Biochemistry, College of Natural Science, Konkuk Univ.