基于ITS2序列的禹州漏芦和漏芦药材基因识别

目的：应用ITS2序列鉴别禹州漏芦和漏芦药材的真伪，为临床准确用药提供可靠的基因识别方法。方法：收集多基原药材禹州漏芦基原物种蓝刺头和华东蓝刺头共14份样品和漏芦药材基原物种祁州漏芦8份样品，经实验获取ITS2序列，结合GenBank中20条相关近缘混伪品序列，建立以上物种和混伪品的DNA条形码鉴定方法；将从市场上收集的11份禹州漏芦和20份漏芦药材的ITS2序列与以上序列进行比对分析和构建系统NJ树。结果：禹州漏芦两个基原物种的序列主导单倍型相同；禹州漏芦和漏芦药材基原物种最大K2P距离均小于其与混伪品的种间最小K2P距离；系统NJ树分析均能准确区分禹州漏芦和漏芦药材与各自混伪品。药材检测结果表明，11份禹州漏芦中10份为正品，1份为伪品；20份漏芦中2份为正品，18份为伪品。结论：本研究建立了基于ITS2序列的禹州漏芦和漏芦药材DNA 条形码鉴定方法，可用于快速鉴别禹州漏芦和漏芦药材真伪。

Identification of Echinopsis Radix and Rhapontici Radix Using ITS2 Sequences

This study aimed to identify Echinopsis Radix and Rhapontici Radix using ITS2 sequences,in order to provide a reliable method for clinical accurate medication by gene recognition.Fourteen samples of Echinops latifolius Tausch and E.grijisii Hance from the origin-species of multi origin traditional Chinese medicine Echinopsis Radix,and eight samples of Rhaponticum uniflorum(L.)DC from the origin-species of Rhapontici Radix,were collected.ITS2 sequences were acquired based on standard experiments,while DNA barcoding system for the materials was established through combining twenty sequences of relative adulterants from GenBank.Based on DNA barcoding system,we detected eleven samples of Echinopsis Radix and twenty samples of Rhapontici Radix from the market.As a result,the sequences of two origin-species of Echinopsis Radix shared the same dominant haplotype.The maximum intra-specific K2P genetic distance between Echinopsis Radix and Rhapontici Radix was less than the minimum inter-specific K2P genetic distance.The analysis of intra-specific and genetic divergence and NJ tree showed that Echinopsis Radix and Rhapontici Radix were separated from their adulterants obviously.Among eleven samples of Echinopsis Radix,ten of them were genuine while the remaining one was fake.However,among twenty samples of Rhapontici Radix,two were genuine and the remaining eighteen were fake.It was concluded that,in this study,the DNA barcoding technique for identification of Echinopsis Radix and Rhapontici Radix was successfully established on the basis of ITS2 sequences,which was a powerful tool in distinguishing Chinese medicines in the marketplaces.