Korean Journal of Plant Resources (한국자원식물학회지)

The Plant Resources Society of Korea (한국자원식물학회)
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Analysis of Genetic Relatedness by Random Amplified Polymorphic DNA (RAPD) in Pecan Taxa

RAPD를 이용한 Pecan 품종의 유전적 관계 분석

  • Published : 2000.03.01
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Abstract

Pecan is deciduous tree and belongs to the Julandaceae family. Pecan is an economically important as a nut and timber crop. Heterozygosity is expected to be high for typically cross-pollinated. Yet little is known about the nature of genetic variation within this species. In addition, the pedigree of many pecan cultivars remains unknown or is questionable. In this study, the phylogenetic relationships between 22 pecan cultivars and its analyzed by RAPD (randomly amplified polymorphic DNA). PCR Amplification used 40 randomly selected oligoes as primers. Based on their genetic similarities derived from the RAPD data, the 22 pecan cultivars were classified into different five groups in agarose gel. The 22 pecan cultivars were classified into five sectional groups by UPGMA clustering analysis, too. C. flacra and Black walnut showed the 0.9 of similarity index and Farley, Pawnee showed the 0.85 of similarity index. The 22 pecan cultivars were classified into different five groups by analysis of the 4% polyacrylamide gel fraction. (Group I : 1, 2, 3, 4, 13, 16, 17, 20, 21 Group II : 14,18 GroupIII : 6,12 GroupIV : 5, 11, 15, 19, 22 CroupV : 7, 8, 9, 10) Group V show the 1.0 of similarity index and Farley, Sturya, Clarke, Pawnee show the 0.98 of similarity index and Kiowa, Schley show the 0.92 of similarity index. Results from this study indicated that RAPD can be used to establish the genetic relationships among the 22 pecan cultivars. Similarity coefficients generally agreed with what would be predicted in cultivars with known pedigrees, and we could accurately construct relationships among cultivars. In addition, we have shown that RAPD provides useful information on the origin of unknown cultivars.

재배되고 있는 22 페칸종간의 RAPD분석을 한 결과, Agarose gel상에서 primer당 평균 6개의 DNA단편이 증폭되었다. 증폭된 DNA 단편의 크기 는 100bp에서 1500bp의 범위에 위치하였다. 페칸종간의 유연 관계를 분석하기에 적합한 primer를 선발한 결과 OPA-02, OPA-10, OPA-12, OPB-01이 최적이었다. 4종의 primer를 이용하여 증폭한 PCR산물을 1.5% Agarose gel상에 분획하여 단편의 종류를 분석한 결과 22의 페칸종은 5개의 그룹으로 구분 할 수 있었다. 40종 primer로부터 증폭된 466 DNA단편을 기초로하여 유전적 근연계수를 계산하였던 결과 유사도 값이 가장큰 것은 C. flacra와 Black walnut로 0.90를 나타났으며, Kiowa와 Red hickory, C. tomentosa, C. flacra, Black walnut간에는 유사도 값이 0.31로 가장 작았다. 선발된 4종의 primer를 이용하여 증폭시킨 PCR산물을 Polyacrylamide gel상에 분획한 후 검출된 DNA단편을 분석하였다. 유사도 값이 가장 큰 것은 그룹 V의 Red hickory, C. tomentosa, C. flacra, Black walnut간으로 1.0이었고, Farley와 Pawnee간과 Sturya와 Clarke간은 공히 0.98를 나타내었다. Polyacrylamide gel분획 후 은염색하여 단편을 검출하는 것은 Agarose gel에 비하여 시간, 기술, 경비가 요구되지만 보다 정확한 유전적 배경을 설명할 수 있다고 생각되었다. 또한 Agarose gel 분석, Polyacrylamide gel 분석 및 주성분 분석법에서 Farley, C. tomentosa, C. flacra, Black walnut, C. corpiformis만이 동일 그룹에서 이탈되지 않았다.

Keywords

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