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该研究选用ITS2序列对升麻及其混伪品进行分子鉴定,从而确保用药安全。实验采用DNA条形码技术,对升麻及其混伪品的ITS2核基因片段进行扩增并双向测序,经CodonCode Aligner V3.7.1拼接后,利用MEGA 5.0软件对相关数据分析,构建邻接(NJ)系统聚类树进行鉴定分析。结果表明升麻药材的3个基原物种:大三叶升麻Cimicifuga heracleifolia、升麻C.foetida、兴安升麻C.dahurica,其ITS2序列长度分别为217,219,219 bp,3个基原物种ITS2序列与混伪品的种间平均K2P距离大于其种内平均K2P距离,ITS2系统发育树显示,升麻的3个基原物种均各自聚为一单系分支,并与其他混伪品明显分开。ITS2序列能够有效准确地鉴别中药材升麻及其混伪品。
In this study, ITS2 sequences were used for molecular identification of cohosh and its adulterants to ensure medication safety. In the experiment, DNA barcoding technique was used to amplify and bi-direction ITS2 gene fragment of cimicifuga and its adulterants. After splicing by CodonCode Aligner V3.7.1, the data were analyzed by using MEGA 5.0 software to construct the adjacent (NJ) system Cluster tree identification analysis. The results showed that three base species of Cimicifuga heracleifolia Cimicifuga heracleifolia, C. foetida and C. dahurica were the 217th, 219th and 219th bp, respectively. ITS2 sequences of three base species The average K2P distance of interspecific hybrids was greater than their average intraspecific K2P distance. The ITS2 phylogenetic tree showed that all three basic species of cohosh were clustered into a single branch and distinctly separated from other adulterants. ITS2 sequence can effectively and accurately identify Cimicifuga and its adulterants.