基于DNA条形码的岭南特色药材广东海桐皮、木棉花与其混淆品的分子鉴定  

Molecular Identification of Erythrina variegate Linn., Bombax malabaricum and Their Adulterants Using DNA Barcode

作　　者：黄娟[1] 徐文[1] 谭瑞湘 白俊其 李西文[2] 黄志海[1] 

机构地区：[1]广州中医药大学第二临床医学院,广州510006 [2]中国中医科学院中药研究所,北京100700

出　　处：《世界科学技术-中医药现代化》2016年第8期1408-1412,共5页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：广东省中医院院内专项(2015KT1817):岭南中草药DNA条形码分子鉴定和生态适宜性研究;负责人:黄志海

摘　　要：目的:本研究主要应用第二内转录间隔区(ITS2)序列作为DNA条形码进行分析,建立对岭南中药广东海桐皮、木棉花与其混淆品的快速鉴定法。方法:收集广东海桐皮等9个物种样本,提取样本基因组DNA,扩增ITS2基因片段,对其产物进行双向测序。所得序列采用Codon Code Aligner进行拼接。利用MEGA 6.06软件进行种间变异及遗传距离分析,并构建系统进化树。结果:9个物种的ITS2序列长度范围为224-237 bp,物种的种内遗传距离(0.000-0.017)明显小于种间遗传距离(0.045-0.820),NJ和ML聚类树显示,各物种样本独聚一支,表现出单系性。结论:ITS2序列能够有效鉴别植物海桐皮、木棉花与其混淆品,为其基原植物鉴定提供依据,为保证用药真实及其临床疗效提供分子鉴定方法。A molecular identification method taken ITS2 as DNA barcode was adopted to distinguish E. variegate, B. malabaricum from their adulterants. Samples from nine species were gleaned. The ITS2 regions were amplified with bi-directional sequencing. Along with some downloaded ones from the GenBank database, all sequences were assembled by CodonCode Aligner and analyzed by MEGA 6.06. The Kimura 2-Parameter （K2P） distances were computed and phylogenetic tree was built using the neighbor-joining （N J） and maximum likelihood （ML） trees. As a result, it was found that the lengths of ITS2 sequence of the nine species varied from 224 to 237 bp. The intraspectific distance （0.000-0.017） was significantly shorter than the interspecific distance （0.045-0.820）. The results of NJ tree and ML tree presented that all the nine species were separately and obviously clustered to one branch, a monophyletic clade. In conclusion, the ITS2 sequence can be treated as an ideal DNA barcode for the molecular identification of E. variegate, B. malabaricum and their adulterants, providing a reference for clinical application.

关 键 词：海桐皮 木棉花 ITS2序列 分子鉴定 DNA条形码 

分 类 号：R282[医药卫生—中药学]