中药材半夏及其混伪品的DNA条形码鉴定研究

摘要

In this study, the ITS2 sequence was used to identify Pinelliae Rhizoma and its adulterants to ensure its market circulation, clinical effect and safety. All genomic DNA from 59 samples were extracted successfully. The Kimura 2-Parameter (K2P) distances and NJ tree were calculated using software MEGA 6.0. The length of the ITS2 sequence of Pinelliae Rhizoma was 251 bp. The intraspecific genetic distance was smaller than the interspecific ones;The NJ tree indicated that Pinelliae Rhizoma distinguished from its adulterants obviously. The results showed that the ITS2 sequence can be used to distinguish Pinelliae Rhizoma from its adulterants accurately and stably.%目的：采用ITS2序列对中药材半夏及其混伪品进行DNA条形码鉴定研究，为规范中药材半夏的市场流通，保障临床用药安全及疗效提供参考依据。方法：对半夏及其混伪品共59份样品，通过DNA提取及聚合酶链式反应（PCR）扩增其ITS2序列，并采用Mega6.0软件进行多序列比对，计算种内及种间K2P遗传距离，采用邻接（NJ）法构建NJ系统聚类树。采用相似性搜索法、最小距离法、NJ树法考察ITS2序列的鉴定能力。结果：半夏药材的ITS2序列长度为251 bp，应用相似性搜索法表明ITS2序列能够准确鉴定半夏药材及其混伪品；半夏种内最大K2P距离小于半夏与混伪品间的最小K2P距离；NJ树显示半夏药材可与其混伪品明显分开。结论：ITS2序列能准确、稳定鉴定中药材半夏药材及其混伪品。