摘要
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。
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.
出处
《世界科学技术-中医药现代化》
2016年第2期202-208,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
科学技术部国家重大新药创制科技专项子课题(2014ZX09304307001-021):中药新药安全性检测技术与标准研究-中药原料药混伪品分子基因鉴定技术与标准
负责人:胡志刚
湖北省科技支撑计划(研发与示范类)(2015BCA275):茯苓等8种湖北优势中药材DNA条形码分子鉴定体系构建
负责人:胡志刚
关键词
禹州漏芦
漏芦
ITS2序列
基因识别
近缘混伪品
Echinopsis Radix
Rhapontici Radix
ITS2 sequences
gene recognition
adulterants
