基于ITS2条形码鉴定水红花子及其混伪品  

Identification of Polygoni Orientalis Fructus and its adulterants using ITS2 barcode

作　　者：任莉[1] 辛天怡[1] 郭梦月 姚辉[1] 庞晓慧 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193

出　　处：《世界中医药》2016年第5期781-785,共5页World Chinese Medicine

基　　金：重大新药创制国家科技重大专项"中药新药安全性检测技术与标准研究"(编号:2014ZX09304307001);国家科技支撑计划(编号:2011BAI07B08)

摘　　要：目的：利用ITS2条形码对中药材水红花子及其混伪品进行鉴定研究。方法：为对该中药材进行准确鉴定,共收集71份样本,包含药材正品及其混伪品。通过对样品进行DNA提取、PCR扩增、双向测序,利用Codon Code Aligner软件进行序列质量评价与拼接,获得ITS2序列。用MEGA软件进行序列比对、变异位点及遗传距离分析,采用最近距离法和构建NJ（邻接）树法来评价ITS2条形码的鉴定能力。结果：水红花子药材基原物种红蓼ITS2序列种内遗传距离为0-0.0124,与其混伪品水蓼、酸模叶蓼以及春蓼的种间遗传距离分别为0.0334-0.0508、0.0688-0.0875、0.0379-0.0467。红蓼种内最大遗传距离小于其与混伪品的种间最小遗传距离,表明ITS2条形码可以准确鉴定水红花子与其混伪品。此外,基于ITS2序列构建的NJ树也可将水红花子及其混伪品明显区分开。结论：ITS2条形码是鉴别水红花子药材的有效工具,可为保障该药材生产投料安全提供新的技术手段。Abstract Objective： The ITS2 barcode was used to identify Polygoni Orientalis Fructus and its adulterants in this study. Methods：For the accuracy of the identification, a total of 71 samples from Polygoni Orientalis Fruetus and its adulterants were collected. The DNA was extracted from the samples and the ITS2 regions were amplified and sequenced. The sequences were then assessed and assembled using the CodonCode Aligner. The genetic distances and variable sites of ITS2 region were analyzed u- sing MEGA after sequence alignment. Nearest distance and phylogenetic tree（NJ-tree） methods were used to test the identification efficiency of the ITS2 barcode. Results：The intraspecifie genetic distances of Polygonum orientale were 0 - 0. 0124, while the interspecific genetic distances between P. orientale and its adulterants were 0. 0334 - 0. 0508,0. 0688 - 0. 0875 and 0. 0379 - 0. 0467, respectively. The maximum intraspecifie genetic distance of P. orientale was lower than the minimum interspecifie genetic distance between P. orientale and its adulterants, which showed that ITS2 bareode could discriminate P. orientale and its adulterants accurately. Besides, the NJ tree based on ITS2 sequences supported that P. orientale and its adulterants can be easily differentiated. Conclusion：The ITS2 bareode is an effective tool for the identification of P. orientale and its adulterants. Our study may provide a new technique to ensure the production feeding security of traditional Chinese medicine.

关 键 词：水红花子 DNA条形码 ITS2 分子鉴定 

分 类 号：R282.5[医药卫生—中药学]