ITS2序列鉴定多基原药材老鹳草  

Identification of Geranii Herbae Using ITS2 Sequence

作　　者：贺海波[1,2] 熊超[1,2] 郭力城[2,3] 凃媛 初旸[1] 汪波 李丹平 孙伟[1] 胡志刚[1,2] 

机构地区：[1]中国中医科学院中药研究所,中药鉴定与安全性检测评估北京市重点实验室,北京100700 [2]湖北中医药大学药学院,武汉430065 [3]广西民族医药研究院,南宁530001 [4]湖北省食品药品监督检验研究院,武汉430064

出　　处：《中国药学杂志》2015年第17期1505-1511,共7页Chinese Pharmaceutical Journal

基　　金：重大新药创制国家重大科技专项(2014ZX09304307001-021);中药新药安全性检测技术与标准研究;中国中医科学院自主课题(Z2013363-865)

摘　　要：目的应用ITS2序列检测市场收集的多基原药材老鹳草,为规范药材市场管理提供分子依据。方法收集多基原药材老鹳草的基原物种牻牛儿苗、老鹳草和野老鹳草及其近缘混伪品,通过实验获取其ITS2序列,结合Gen Bank下载的序列共计28个物种82条序列,进行序列比对验证、最近距离法和系统邻接(NJ)树分析,建立基于ITS2序列的老鹳草药材DNA条形码鉴定技术方法;运用该方法检测市场随机收集的18份老鹳草药材样品,经序列比对结合以上方法建立的系统邻接(NJ)树确定其物种基原,以判定其市场真伪。结果 ITS2序列能将老鹳草药材的3个基原物种及其近缘混伪品很好地区分开;18份市场收集的老鹳草药材ITS2序列鉴定结果表明,其中16份为正品,2份为混伪品。结论该研究建立了基于ITS2序列的多基原药材老鹳草DNA条形码鉴定技术方法,实现了对市场随机收集的老鹳草药材物种基原的真伪判定,为多基原药材鉴定难题提供了参考方法。OBJECTIVE： To provide the molecular basis for standard medicine market management, a new method to identify multi-origin traditional Chinese medicine Geranii herba was established by ITS2 sequence analysis. METHODS As a prelude to species identification, Erodium stephaniahum willd. , Geranium carolinianum L. , Geranium wilfordii Maxim. , together with their relatives adulterants, were sampled. A reference ITS2 sequence library was developed, including 82 ITS2 sequences of 28 species download from GenBank. Based on the intra-species, inter-species divergence, variable sites and the neighbor-joining （NJ） phylogenetic tree observed, a molecular identification method was established to the multi-origin herbs Geranium using ITS2 sequence. RESULTS The NJ trees showed three origin species formed one monophyletic clade respectively; in addition any one of multi-origin herbs Geranium and its relatives adulterants could be distinguished clearly. Using this method, 18 random copies of Geranium herb collected from market were analyzed, 16 of which are genuine, 2 as adulterants. CONCLUSION Therefore ITS2 sequence is suitable to be as a barcode to identify multi-origin herbs Geranium and their relatives adulterants.

关 键 词：老鹳草 ITS2序列 鉴定 近缘混伪品 

分 类 号：R282[医药卫生—中药学]