Journal of Mushroom (한국버섯학회지)

The Korean Society of Mushroom Science (한국버섯학회)
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Development of strain-specific SCAR marker for selection of Pleurotus eryngii strains with higher β-glucan

베타글루칸 함량이 높은 큰느타리버섯 선발을 위한 SCAR marker 개발

  • Kim, Su Cheol (Department of Pharmaceutical Engineering, Gyeongnam National University of Science and Technology) ;
  • Kim, Hye Soo (Department of Pharmaceutical Engineering, Gyeongnam National University of Science and Technology) ;
  • Cho, Yong Un (Department of Pharmaceutical Engineering, Gyeongnam National University of Science and Technology) ;
  • Ryu, Jae-San (Eco-friendliness Research Department, Gyeongnam-do Agricultural Rearch and Extension Services) ;
  • Cho, Soo Jeong (Department of Pharmaceutical Engineering, Gyeongnam National University of Science and Technology)
  • 김수철 (경남과학기술대학교 제약공학과) ;
  • 김혜수 (경남과학기술대학교 제약공학과) ;
  • 조용운 (경남과학기술대학교 제약공학과) ;
  • 류재산 (경상남도농업기술원 친환경연구과) ;
  • 조수정 (경남과학기술대학교 제약공학과)
  • Received : 2015.03.09
  • Accepted : 2015.03.30
  • Published : 2015.03.31
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Abstract

In this study, SCAR marker that differentiates Pleurotus eryngii strains with higher ${\beta}$-glucan from control strain was developed. Genomic DNAs of 9 control strains of Pleurotus eryngii and 9 Pleurotus eryngii strains with higher ${\beta}$-glucan were analyzed by bulked segregant analysis (BSA) using randomly amplified polymorphic DNA (RAPD). One-hundred twenty RAPD primers were screened on bulked DNA samples and a unique DNA fragment with the size of 91 bp was yielded by OP-R03 primer from the Pleurotus eryngii strains with higher ${\beta}$-glucan. A sequence characterized amplified region (SCAR) marker, designated as OP-R03-1-F and OP-R03-1-R, was designed on the basis of the determined sequence. The PCR analysis with the OP-R03-1 primer showed that this SCAR marker can clearly distinguish the Pleurotus eryngii strains with higher ${\beta}$-glucan from the control strains.

본 연구는 큰느타리버섯의 베타글루칸 고함유 형질에 관련된 SCAR marker를 개발하기 위해 수행되었다. operon사의 OPA(20개), OPB(20개), OPL(20개), OPP(20개), OPR(20개), OPS(20개) 등 총 120개 primer를 random primer(10 mer)로 사용하여 대립 계통 9종과 베타글루칸 고함유 계통 9 종을 대상으로 RAPD를 이용한 bulked segregant analysis를 실시하여 OP-R03 primer로부터 대립 계통에는 나타나지 않고 베타글루칸 고함유 계통에만 나타나는 특이적인 RAPD 밴드를 얻었다. OP-R03 primer를 이용한 RAPD 결과, 약 91 bp 부근에서 베타글루칸 고함유 계통에 특이적인 DNA 밴드가 관찰되었으며 이 DNA 밴드의 염기서열 말단을 근거로 SCAR 마커로 사용할 specific primer인 OP-R03-1-F와 OP-R03-1-R를 디자인하였다. SCAR 마커 OP-R03-1-F/-1-R primer를 이용하여 PCR을 수행한 결과에서도 91 bp 부근에서 대립 계통과 구별되는 DNA 밴드가 베타글루칸 고함유 계통에서 확인되었으며 random primer인 OP-R03 primer를 이용하여 PCR을 수행했을 때보다 재현성이 높고 진한 DNA 밴드임을 확인할 수 있었다.

Keywords

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