常见药用蛇类的DNA条形码研究  

DNA Barcoding of Common Medicinal Snakes In China

作　　者：廖婧[1] 梁镇标[1] 张亮[2] 李军德[3] 晁志[1] 

机构地区：[1]南方医科大学中医药学院,广州510515 [2]华南濒危动物研究所,广州510260 [3]中国中医科学院中药研究所,北京100700

出　　处：《中国药学杂志》2013年第15期1255-1260,共6页Chinese Pharmaceutical Journal

基　　金："重大新药创制"科技重大专项(2009ZX09301-005-03);广东省暨国家级大学生创新创业训练计划项目(20122121056)

摘　　要：目的建立国内常见药用蛇类的DNA条形码数据库，用于生药样品鉴定并分析物种间的亲缘关系。方法收集常见药用蛇类23种，共44个样品，对各样品COI条形码序列进行测定。所得序列录入数据库；用MEGA5．0等软件进行遗传距离等分析，构建NJ树，分析各物种的种内变异，种间变异，以及系统树中各物种的聚类情况。结果各物种的种间存在277个变异位点，种内遗传距离值范围为0～0．0409，种间遗传距离值范围为0．0850—0．2568，种内遗传距离与种间遗传距离有显著差异；由所构建的系统聚类树图可以看出，样品可聚类为与分科相一致的3个类群，各物种均形成了相对独立的支。结论COI条形码能够对实验中各药用蛇类物种进行准确鉴定，具有科学、准确、简便等特点，可以推广运用于其他蛇类的鉴别；研究还为上述蛇类物种间亲缘关系提供了新的参考信息。OBJECTIVE To establish a DNA barcode database of common medicinal snakes in China for species identification and phylogenetic analysis. METHODS The COl barcode sequences of 44 samples from 23 snake species were amplified and sequenced. The aligned sequences, 658 base pairs in length, were analyzed for divergence using the Kimura-2-parameter distance model with MEGA5.0. RESULTS The genetic distance and NJ tree demonstrated that there were 277 variable sites in species, the intraspecific K2P distance ranged from 0 - 0. 040 9, the interspecific genetic distance ranged from 0. 085 0 - 0. 256 8, and there was significant difference between the interspecific genetic distance and intraspecific genetic distance. The minimum interspecific distance was 0. 085 0 between Lycodon synaptor and Lycodon ruhstrati. The maximum interspecific distance was 0. 256 8 between Elaphe radiate and Naja atra. The snakes examined in this study were clustered in three major elades corresponding to their three families. Each species formed a monophyletic clad. CONCLUSION DNA barcoding based on the COI sequences is accurate and reliable and could be applied to identify the common medicinal snakes in our study. The study provides new information for the phylogenetic relationships of snake species.

关 键 词：COI基因 蛇类 DNA条形码 鉴定 

分 类 号：R284[医药卫生—中药学]