薄荷、留兰香特异性引物PCR鉴别方法研究  

Molecular Identification of Mentha haplocalyx and Mentha spicata with Specific Primers Multi-PCR System

作　　者：曹亮[1,2,3] 秦双双[4] 袁媛[2] 朱校奇[1] 

机构地区：[1]湖南省农业科学院农业生物资源利用研究所,湖南长沙410125 [2]中国中医科学院中药资源中心,北京100700 [3]湖南中医药大学药学院,湖南长沙410208 [4]广西药用植物园,广西南宁530023

出　　处：《中药材》2014年第1期41-45,共5页Journal of Chinese Medicinal Materials

基　　金：公益性行业(农业)科研专项(201303117);北京市科技项目“道地中药功能基因组北京市重点实验室2012年阶梯计划项目”

摘　　要：目的:筛选留兰香、薄荷鉴别SNP位点,并设计特异性引物,实现薄荷、留兰香及二者混合品的快速鉴别。方法:通过测序及GenBank中下载获得薄荷、留兰香的psbA序列,通过使用Bioedit软件比对获得SNP位点,设计特异性多重PCR引物,通过条件优化,对薄荷、留兰香原植物及药材进行鉴别。结果:构建了多重PCR鉴别体系,能扩增出薄荷181 bp的鉴别条带或(和)留兰香288 bp鉴别条带,通过一个PCR反应就能对药材及二者混合品进行鉴别。结论:使用多重PCR技术可以有效鉴别薄荷、留兰香及二者的混合品。Objective：To screen specific SNPs loci of Mentha haplocalyx and Mentha spicata,and then specific primers were designed to identify the two species and their mixture rapidly.Methods：PsbA-trnH sequences of Mentha haplocalyx and Mentha spicata were obtained by PCR product sequencing and downloading from GenBank.SNPs in the psbA-trnH sequences of Mentha haplocalyx and Mentha spicata were found by ClustulW program and Bioedit software.Primers for authentication of the two species were designed according to the SNP loci,and PCR reaction system was optimized to identify the original plants.Results：Multi-PCR reaction system was constructed.The 181 bp identification band for Mentha haplocalyx or（ and） 288 bp identification band for Mentha spicata could be produced by a single PCR reaction,which showed good identification ability to the two species.Conclusion：The multi-PCR reaction system can be applied to identify Mentha haplocalyx and Mentha spicata as well as their mixture.

关 键 词：薄荷 留兰香 SNP PCR 分子鉴别 

分 类 号：R282.5[医药卫生—中药学]