ALM-ASA法鉴别人参和西洋参ITS及5．8s基因上单核苷酸多态性位点

Identification of SNP on ITS and 5.8s gene of Panax Ginseng and P.quinquefolium by ALM-ASA

目的:查找并发现参类药材的单核苷酸多态性(SNP)位点,利用SNP的准确测定来鉴别人参和西洋参。方法:根据ITS及5.8s基因上人参和西洋参的SNP位点设计特异性引物,利用适配器连接介导的等位基因特异性扩增法(ALM-ASA)进行检测。结果:PCR反应体系优化后,能在同一反应中同时鉴别人参、西洋参。根据凝胶电泳中扩增片段的大小判断SNP的类型,出现294bp条带的为人参,111bp条带的为西洋参。结论:ALM-ASA法极大提高了PCR反应的特异性,结果准确,可同时测定多个SNP位点。采用该方法测定ITS及5.8s基因上的SNP位点能有效地对参类品种进行鉴别和质量控制。

Aim： To identify single nucleotide polymorphisms （SNP） sites in Panax species and to establish an accurate method for the identification of P. ginseng and P. quinquefolium. Methods： Specific primers for P. ginseng and P. quinquefolium were designed based on the SNP on ITS and 5.8s gene. Adapter-ligation mediated allele-specific amplification（ALM-ASA） method was used to detect the type of SNP. Results： After optimizing the PCR reaction system, the template DNA of P. ginseng was amplified into 294 bp band whereas P. quinquefolium into 111 bp. The results showed that the method was specific and reproducible and could inplement the detection in one tube. Conclusion： ALM-ASA technique markedly enhanced the specificity of PCR. It can be used to simultaneously type multiple SNPs. SNP typing of ITS and 5.8s gene is suggested as a criterion for its identification of Panax species, and also used as an evaluation method for its quality control.