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本研究基于NCBI数据库中文心兰的EST序列,对其EST-SSR标记进行分析,并设计了20对EST-SSR引物用于30个文心兰品种遗传多样性性分析研究。对3 183条文心兰EST序列进行搜索,共检索出274个SSR位点,检出率为8.61%。二核苷酸重复类型是文心兰EST-SSR的主要重复类型,占60.58%。通过PCR扩增检测到13对引物有扩增产物,其中10对引物有多态性,平均每对引物扩增到10个多态性条带,各引物的多态性比率分布为75.00%~100.00%,多态性信息含量变化范围为0.69~0.93。聚类分析发现30个文心兰品种间的遗传距离变化在0.04~0.58,且在遗传距离为0.39处可将30个文心兰品种分为7大聚类群。
In this study, EST-SSR markers were analyzed based on the EST sequences of A. annua in NCBI database, and 20 pairs of EST-SSR primers were designed for genetic diversity analysis of 30 species of Oncidium. A total of 274 SSR loci were found from 3 183 ESTs of A. sinensis and the detection rate was 8.61%. The dinucleotide repeat type is the major repeat type of EST-SSR, accounting for 60.58%. Thirteen pairs of primers were amplified by PCR, of which 10 pairs of primers were polymorphic, with an average of 10 polymorphic bands per pair of primers. The polymorphism rate of each primer was 75.00% 100.00%. The content of polymorphism information varied from 0.69 to 0.93. Cluster analysis showed that the genetic distance among 30 species of Oncidium was 0.04-0.58, and 30 species were divided into 7 clusters at the genetic distance of 0.39.