Journal of Korean Society of Forest Science (한국산림과학회지)

Korean Society of Forest Science (한국산림과학회)
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Development and Characterization of Chloroplast Simple Sequence Repeat markers in Pinus koraiensis

잣나무 엽록체 Simple Sequence Repeat 표지자 개발 및 특성 분석

  • Lee, Jei-Wan (Division of Forest Genetic Resources, National Institute of Forest Science) ;
  • Baek, Seung-Hoon (Division of Forest Genetic Resources, National Institute of Forest Science) ;
  • Hong, Kyung-Nak (Division of Forest Genetic Resources, National Institute of Forest Science) ;
  • Hong, Yong-Pyo (Division of Forest Genetic Resources, National Institute of Forest Science) ;
  • Lee, Seok-Woo (Division of Forest Genetic Resources, National Institute of Forest Science) ;
  • Ahn, Ji-Young (Division of Forest Genetic Resources, National Institute of Forest Science)
  • 이제완 (국립산림과학원 산림유전자원과) ;
  • 백승훈 (국립산림과학원 산림유전자원과) ;
  • 홍경낙 (국립산림과학원 산림유전자원과) ;
  • 홍용표 (국립산림과학원 산림유전자원과) ;
  • 이석우 (국립산림과학원 산림유전자원과) ;
  • 안지영 (국립산림과학원 산림유전자원과)
  • Received : 2015.07.10
  • Accepted : 2015.08.10
  • Published : 2015.12.31
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Abstract

Novel cpSSR primers were developed based on the sequence information of the Pinus koraiensis chloroplast genome. A total of 30 cpSSR loci were detected in the chloroplast genome, and a total of 30 primer sets flanking those loci were designed. All primer sets were successfully amplified for chloroplast DNA in P. koraiensis. The cross-species transferability of the 30 primer sets was considerably high in P. pumila (100%) and P. paviflora (97%) belonging to the same Subgenus (Strobus) of P. koraiensis. Meanwhile, the transferability was relatively low (73%) in P. densiflora and P. sylvestris belonging to Subgenus Pinus. A total of 13 cpSSR loci out of the 30 loci were polymorphic in the Mt. Jumbong population of P. koraiensis. The mean of haploid diversity(H) was 0.512. The number of haplotypes(N) and the haplotype diversity($H_e$) were 25 and 0.992, respectively. Of the 25 haplotypes, 22 were unique in the analyzed population. The unique haplotypes differentiated 22 individuals (79%) from the total of 28 individuals. In conclusion, the novel cpSSR primers developed in this study would be applicable to other Pinus species, especially the subgenus Strobus, and provide a high level of polymorphism for the study of genetic variation of P. koraiensis.

본 연구에서는 잣나무 엽록체 DNA의 전체 염기서열을 기반으로 엽록체 SSR(chloroplast simple sequence repeat) 영역을 특이적으로 증폭하는 primer를 개발하고 그 특성을 분석하였다. 잣나무 엽록체 DNA에서 총 30개의 SSR 영역을 탐색하였으며, 이들 영역을 증폭하기 위한 30개의 primer를 제작하였다. 모든 primer가 잣나무를 대상으로 PCR 증폭이 가능하였다. 근연종에 대한 primer의 종간 전환률은 잣나무와 동일한 아속(Subgenus Strobus)에 속하는 눈잣나무(100%)와 섬잣나무(97%)에서 가장 높게 나타났다. 반면 소나무아속(Subgenus Pinus)에 속하는 소나무와 구주소나무에서의 종간 전환률은 73%로 비교적 낮게 나타났다. 점봉산 잣나무 집단을 대상으로 조사한 결과 13개의 유전자좌에서 다형성이 관찰되었으며, 평균 haploid 다양도(H)는 0.512로 계산되었다. 다형적 유전자좌로부터 조합된 haplotype의 수(N)는 25개로 확인되었고, haplotype 다양도($H_e$)는 0.992로 매우 높게 나타났다. 집단내 독특하게 관찰되는 haplotype은 22개(88%)로 전체 28개체 중에서 22개체(79%)를 식별하였다. 본 연구에서 개발한 cpSSR primer는 높은 종간 전환률을 나타냄에 따라 소나무속의 근연종, 특히 잣나무아속 수종에 활용 가능성이 높고, 잣나무 유전변이 분석을 위한 충분한 다형성을 제공하는 유용한 표지자로 판단된다.

Keywords

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