基于SNP的多重位点特异性PCR鉴别不同居群竹节参

Identification of different populations of Panax japonicus by multiple allele-specific polymerase chain reaction

竹节参,五加科人参属,是具有极高药用价值的珍稀濒危药用植物。但竹节参的居群混杂亟需建立一种能准备鉴别竹节参不同居群来源的方法。基于前期比较转录组数据筛选竹节参SNP位点,并基于这些SNP位点设计特异性引物,并对来自不同居群的竹节参(云南鲁甸、湖北华中药用植物园、四川乐山、陕西平利县、湖北七姊妹山)进行PCR鉴定和优化。结果筛选得到磷酸甲羟戊酸激酶1(PMK1)基因上的A/G点突变是可鉴别不同居群竹节参遗传变异的有效SNP位点。本实验室初步建立了基于竹节参的单核苷酸多态性(SNP)位点的PCR体系,进一步鉴别了不同居群的竹节参,为实现竹节参及其近缘种的分子鉴别奠定基础。

Panax japonicus is a rare and endangered medicinal plant with high medicinal value.However,it is necessary to establish a method to identify the origin of different populations of Panax japonicus.Methods:in this study,SNP loci of Panax japonicus were screened based on the data of pre comparative transcriptome,and specific primers were designed based on these SNP loci.PCR identification and optimization were carried out for Panax japonicus from different populations(Ludian in Yunnan,Huazhong Medicinal in Hubei,Leshan in Sichuan,Pingli County in Shanxi and Mount Qizimei in Hubei).Results:the A/G point mutation in pmk-1 gene was an effective SNP site to identify genetic variation in different populations of Panax japonicus.Conclusion:a PCR system based on SNP locus of Panax japonicus was established in our laboratory,which further identified different populations of Panax japonicus,and laid a foundation for molecular identification of Panax japonicus and its related species.