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目的:探讨霍山石斛的叶绿体基因组(chloroplast genomic DNA,cpDNA)中微卫星分布规律并设计引物鉴别霍山石斛及其伪品。方法:利用GenBank上已公布的霍山石斛cpDNA全序列,应用分子生物学软件对霍山石斛cpDNA中的微卫星位点统计分析,并设计筛选特异性引物对霍山石斛进行鉴别。结果:霍山石斛cpDNA中微卫星位点共有52个,以单碱基重复序列为主,共有25个,占总位点48%,其次二碱基重复序列有16个,占总位点30.8%,三碱基及三碱基以上重复序列在cpDNA中分布极少。在通过微卫星位点所设计的4对引物中,有2对引物可对霍山石斛及部分混伪品进行有效的鉴别。结论:所设计的引物可以对霍山石斛及其伪混品进行鉴别,且所得数据为霍山石斛的种群遗传和起源分化的研究提供了支持。
Objective: To investigate the distribution of microsatellites in chloroplast genomic DNA (cpDNA) of Dendrobium huoshanense and to design primers to identify Dendrobium huoshanense and its counterfeit products. Methods: Using the published sequence of cpDNA from Dendrobium huoshanense in GenBank, the microsatellite loci in cpDNA of Dendrobium huoshanense were analyzed by molecular biology software. Dendrobium huoshanense was identified through screening specific primers. Results: There were 52 microsatellite loci in the dendrobium huoshanense, which were dominated by single base repeats and 25, accounting for 48% of the total loci, followed by 16 with two base repeats, accounting for 30.8% , Three bases and more than three base repeats in cpDNA distribution is minimal. Among 4 pairs of primers designed by microsatellite loci, 2 pairs of primers could effectively identify Dendrobium huoshanense and some mixed products. Conclusion: The designed primers can identify Dendrobium huoshanense and its fake hybrid, and the obtained data provide support for the population genetic and origin differentiation of Dendrobium huoshanense.