한국버섯학회지 (Journal of Mushroom)

  • 제15권2호
  • /
  • Pages.78-83
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  • 2017
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  • 1738-0294(pISSN)
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  • 2288-8853(eISSN)
한국버섯학회 (The Korean Society of Mushroom Science)
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팽이버섯 (Flammulina velutipes) 계통의 분류를 위한 SSR 마커개발

Development of SSR markers for classification of Flammulina velutipes strains

  • 우성이 (농촌진흥청 국립원예특작과학원 버섯과) ;
  • 서경인 (국립종자원) ;
  • 장갑열 (농촌진흥청 국립원예특작과학원 버섯과) ;
  • 공원식 (농촌진흥청 국립원예특작과학원 버섯과)
  • Woo, Sung-I (Mushroom Research Division, National Institute of Horticultural & Herbal Science, Rural Development Administration) ;
  • Seo, Kyoung-In (Korea seed & variety service) ;
  • Jang, Kab yeul (Mushroom Research Division, National Institute of Horticultural & Herbal Science, Rural Development Administration) ;
  • Kong, Won-Sik (Mushroom Research Division, National Institute of Horticultural & Herbal Science, Rural Development Administration)
  • 투고 : 2017.03.15
  • 심사 : 2017.06.20
  • 발행 : 2017.06.30
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초록

버섯과에서 한국, 중국, 일본에서 재배 또는 수집하여 농촌진흥청에 보관 중인 32개의 팽이버섯 계통에 대하여 조사하였다. 팽이버섯의 미소반복서열(microsatellite)을 포함하고 있는 490개의 DNA 단편을 얻었다. 다양한 팽이버섯 균들의 PCR을 통한 DNA 프로파일링을 수행함으로써 다형성 변이가 많이 검출되었다. 총 34개의 대립 유전자가 12개의 다형성 SSR 마커 중에서 검출되었고, 평균 3.42개의 대립 유전자와 대립 유전자의 수는 유전자좌당 2개에서 7개까지 분포하였다. 대립 형질 빈도는 0.42(GB-FV-127)에서 0.98(GB-FV-166)이었으며 이형접합체 관측치($H_O$)와 기대치($H_E$)는 각각 0.00에서 0.94(평균 = 0.18)와 0.03에서 0.67(평균 = 0.32)이었다. 다형성 지수는 (PIC) GB-FV-127 마커에서 가장 높은 0.61, 평균대립 유전자 수는 5를 나타내었고, GB-FV-166마커에서 0.03과 2로 가장 낮았다. 본 연구에서 평균 PIC 값(0.29)은 대립 유전자의 평균 수(3.42)로 관찰되었다. 결론적으로 우리는 풍부한 SSR 라이브러리에서 12 개의 다형성 SSR 마커를 개발하는데 성공했다. 이러한 SSR은 계통 발생 분석, 유전적 변이 평가에 중요하게 사용될 것이다.

Microsatellite SSR markers were developed and utilized to reveal the genetic diversity of 32 strains of Flammulina velutipes collected in Korea, China, and Japan. From the SSR-enriched library, 490 white colonies were randomly selected and sequenced. Among the 490 sequenced clones, 85 (17.35%) were redundant. Among the remaining 405 unique clones, 201 (49.6%) contained microsatellite sequences. We used 12 primer pairs that produced reproducible polymorphic bands for four diverse strains, and these selected markers were further characterized in 32 Flammulina velutipes strains. A total of 34 alleles were detected using the 12 markers, with an average of 3.42 alleles, and the number of alleles ranged from two to seven per locus. The major allele frequency ranged from 0.42 (GB-FV-127) to 0.98 (GB-FV-166), and values for observed ($H_O$) and expected ($H_E$) heterozygosity ranged from 0.00 to 0.94 (mean = 0.18) and from 0.03 to 0.67 (mean = 0.32), respectively. SSR loci amplified with GB-FV-127 markers gave the highest polymorphism information content (PIC) of 0.61 and mean allele number of five, whereas for loci amplified with GB-FV-166 markers these values were the lowest, namely 0.03 and two. The mean PIC value (0.29) observed in the present study with average number of alleles (3.42). The genetic relationships among the 32 Flammulina velutipes strains on the basis of SSR data were investigated by UPGMA cluster analysis. In conclusion, we succeeded in developing 12 polymorphic SSRs markers from an SSR-enriched library of Flammulina velutipes. These SSRs are presently being used for phylogenetic analysis and evaluation of genetic variations. In future, these SSR markers will be used in clarifying taxonomic relationships among the Flammulina velutipes.

키워드

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