基于ITS2序列的矮地茶药材基因识别  

Identification of Ardisiae Japonicae Herba by ITS2 Sequence

作　　者：答国政 张秀桥[1] 姚辉[2] 王志平[1] 赵博[3] 张超[1] 胡志刚[1,3] 

机构地区：[1]湖北中医药大学药学院,湖北武汉430065 [2]中国医学科学院北京协和医学院药用植物研究所,北京100193 [3]中国中医科学院中药研究所,北京100700

出　　处：《中药材》2015年第11期2277-2280,共4页Journal of Chinese Medicinal Materials

基　　金：湖北省科技支撑计划项目(2015BCA275);中国科学院"西部之光"人才培养计划项目[科发人教字(2012)179号]

摘　　要：目的:采用ITS2序列鉴定从市场上收集的矮地茶药材,为保证矮地茶用药安全提供可靠的基因识别方法。方法:收集矮地茶药材基原物种紫金牛及其混伪品共17个物种56份样品,并从NCBI下载6条相关序列,分析上述物种ITS2序列的种内和种间变异位点、Kimura 2-parameter(K2P)距离,以邻接(NJ)法构建系统树,建立基于ITS2序列的矮地茶药材DNA条形码鉴定技术方法。同时,从市场上随机收集15份矮地茶药材样品,通过ITS2序列比对和构建系统聚类树判定药材真伪。结果:紫金牛ITS2序列种内K2P距离最大值(0.0059)和平均值(0.0010)均明显小于其与混伪品种间K2P距离最小值(0.0364)和平均值(0.1159),系统NJ树能将紫金牛与混伪品明显区分开;15份矮地茶药材的序列比对和系统NJ树鉴定结果表明,其中13份为正品,2份为混伪品。结论:本研究建立了基于ITS2序列的矮地茶药材DNA条形码鉴定技术方法,为快速检测市场流通的矮地茶药材真伪提供了可借鉴的基因识别方法。Objective： The ITS2 sequence was used to identify Ardisiae Japonicae Herba collected from the market,in order to ensure the medicine quality of the market and to provide a reliable technical method. Methods： The certified samples,including Ardisia japonica and its adulterants,were 56 samples of 17 species. All the sequences of the samples including six related sequences downloaded from NCBI were analyzed by computing the Kimura 2-parameter（ K2P） genetic distance and the neighbor-joining（ NJ） phylogenetic tree,then the method of DNA barcoding identification technology of Ardisiae Japonicae Herba based on ITS2 sequence was established. Combined with online comparison of sequences,the method was used to detect 15 samples of Ardisiae Japonicae Herba from the market to distinguish authenticity. Results： The maximum and the average intra-specific K2 P genetic distance of Ardisia japonica were all less than the minimum and the average inter-specific K2 P genetic distance,and Ardisia japonica and its adulterants could be separated by computing the NJ phylogenetic tree. The identification results of online comparison and DNA barcoding identification were the same. In all of the 15 samples,13 of them were genuine,and the other two samples were fake. Conclusion： The DNA barcoding identification technique method of Ardisiae Japonicae Herba is established based on ITS2 sequence,and it provides a reference method to distinguish the authenticity of Ardisiae Japonicae Herba quickly by gene recognition.

关 键 词：矮地茶 紫金牛 ITS2序列 基因识别 

分 类 号：R384.1[医药卫生—医学寄生虫学]