基于COI和16SrRNA基因的地龙药材及其混淆品的DNA条形码鉴定  

DNA Barcode Identification of Pheretima Medicinal Material and Its Adulterants Based on COI and16SrRNA Gene Sequence

作　　者：马梅[1] 李薇[1] 龚玲[1] 卢瑞珊[1] 

机构地区：[1]广州中医药大学中药学院,广东广州510006

出　　处：《中药新药与临床药理》2014年第5期595-598,共4页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然基金项目(81374035)

摘　　要：目的利用线粒体细胞色素C氧化酶亚基1(COI)和16S核糖体RNA(16SrRNA)对地龙药材及常见混伪品进行分子鉴定,探讨快速、准确鉴定地龙药材及其混伪品基原物种的方法。方法收集地龙药材的药典收载品种及其易混品种10种,提取DNA,得到其COI和16SrRNA序列。所得序列经DNAStar校正后,用MEGA6.0自带的Clustal W多重比对,除去冗余位点,比对结果经MEGA6.0分析,构建地龙药材及其混淆品的邻接(N-J)树。结果地龙药材的正品来源参环毛蚓(Pheretima aspergillum)、通俗环毛蚓(P.vulgaris)、威廉环毛蚓(P.guillelmi)及栉盲环毛蚓(P.pectinifera)与其混淆品种间COI和16SrRNA基因序列均存在较多变异位点。由所构建的N-J系统聚类树图显示所测物种的单系性,即16SrRNA、COI可以很好的将地龙药材区别于其他混伪品。结论所获得的COI和16SrRNA序列可作为不同状态的地龙类药材物种鉴定的分子依据,并为进一步建立地龙等动物类中药物种真实性鉴定体系奠定了重要的实验基础。Objective COI and 16SrRNA genes were used for the molecular identification of Pheretima medicinal material and its commonly-seen adulterants, thus to build a rapid, accurate method for the identification of their species. Methods We collected 10 authentic and adulterant species of Pheretima medicinal material from the producing region, and then we extracted DNA and obtained the COI and 16SrRNA sequences. DNAStar and MEGA6.0 were applied to analyze the variable sites and construct their N-J tree. Results Authentic Pheretima medicinal material came from the species of Pheretima aspergillum, P. vulgaris, P. guillelmi, and P. pectinifera. Compared with the authentic Pheretima medicinal material, the adulterant had lots of variable sites. Authentic Pheretima medicinal material and its adulterants can be identified by COI and 16SrRNA genes. Conclusion The obtained COI and 16SrRNA genes can provide reference for the identification of Pheretima medicinal material at the molecular level, and lay the important experimental basis for further establishment of authenticity identification of Pheretima as well as other animal medicines.

关 键 词：地龙 线粒体细胞色素C氧化酶亚基1 16S核糖体RNA DNA条形码 

分 类 号：R284.1[医药卫生—中药学]