Journal of Mushroom (한국버섯학회지)

The Korean Society of Mushroom Science (한국버섯학회)
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Identification of Pleurotus ostreatus cultivars with the application of multiplex-simple sequence repeat markers

Multiplex SSR마커를 이용한 느타리(Pleurotus ostreatus) 품종 판별

  • Choi, Jong In (Gyeonggi-do Agricultural Research &Extension Services) ;
  • Jung, Hwa Jin (Department of mushroom science, Korea National college of Agriculture and fisheries) ;
  • Na, Kyeong sook (Department of mushroom science, Korea National college of Agriculture and fisheries) ;
  • Oh, Min-Ji (Mushroom Research Division, National Institute of Horticultural & Herbal Science, RDA) ;
  • Kim, Min-Keun (Gyeongsangnam-do Agricultural Research and Extension Services) ;
  • Ryu, Jae-San (Department of mushroom science, Korea National college of Agriculture and fisheries)
  • 최종인 (경기도농업기술원 버섯연구소) ;
  • 정화진 (한국농수산대학 버섯학과) ;
  • 나경숙 (한국농수산대학 버섯학과) ;
  • 오민지 (농촌진흥청 버섯과) ;
  • 김민근 (경남농업기술원) ;
  • 류재산 (한국농수산대학 버섯학과)
  • Received : 2021.02.26
  • Accepted : 2021.03.26
  • Published : 2021.03.31
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Abstract

To develop a method for the differentiation of Pleurotus ostratus cultivars, the multiplex-simple sequence repeat (SSR) primer set based on the SSRs obtained from whole genomic DNA sequence analysis was designed with two polymerase chain reaction (PCR) primer sets. These SSR primer sets were employed to distinguish 10 cultivars and strains. Twenty polymorphic markers were selected based on the genotyping results. PCR with each primer produced 1-4 distinct bands ranging in size from 150 to 350 bp, which was within the expected range. However, since a sole SSR marker was unable to detect polymorphisms in every cultivar, multiplex PCRs with composite PCR primer sets were employed. The multiplex primer, "166+115," completely discriminated 12 cultivars and strains with 40 loci, which were 12 more than the simple arithmetic addition of each locus of the primers 115 and 166. These results might be useful to provide an efficient method for the differentiation of P. ostreatus cultivars with separate PCRs for the quality control of spawn and protection of breeders' rights.

느타리 품종구분을 위한 마커의 개발을 위하여 곤지7호의 어버이 일핵 균사중의 하나인 MT07156-97의 전체 유전자 염기서열을 바탕으로 제작한 251개의 SSR 프라이머를 제작하였다. 우선적으로 수한1호, 곤지7호, 흑타리 품종에 다형성 여부를 관찰하여 20개의 SSR을 선발하고, 이를 10개 품종에 적용하였다. 단일의 프라이머로는 일부 품종이 구분되지 않았으므로, 선발된 프라이머 간의 다양한 조합(multiplex 방식)을 적용한 결과 모든 품종을 판별할 수 있는 분자마커 다형성을 보인 프라이머 "166+115" 조합을 선발하였다. 별도로 프라이머 115와 166가 만들어 낸 산술적인 유전자좌(loci) 31개보다 12개 많은 40개의 유전자좌가 증폭되어 다양한 품종에 특이적인 분자마커를 제공할 수 있었다. 개발된 분자마커는 종균의 품질관리, 품종의 판별, 신품종 보호에 활용될 수 있을 것이다.

Keywords

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