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目的:研究黄芪与其混伪品之间的DNA分子鉴别方法。方法:采集不同产地的黄芪药材13份,替代品红芪2份,混伪品紫花苜蓿3份和蜀葵1份,所有样品进行总DNA的提取,PCR扩增,并对扩增产物进行测序得到相应的序列,同时从GenBank下载蓝花棘豆、锦鸡儿2种伪品的ITS序列。用MEGA 4计算其种间的K-2-P距离,最后利用ITS序列构建其系统发育树。结果:测得了19份样品的ITS序列全长,分别为蒙古黄芪646～650 bp,膜荚黄芪为646～650 bp;红芪为664 bp;紫花苜蓿为659 bp;蜀葵为728 bp,在GenBank中注册,获得登记号。通过以ITS序列重建系统进化树进行的聚类分析可以将黄芪与其混伪品有效的区分开。结论:ITS序列能够成功鉴定黄芪及其易混伪品,可以作为黄芪与其混伪品的分子鉴定方法。
Objective: To study the molecular identification of DNA between Astragalus and its adulterants. Methods: Thirteen samples of astragalus membranaceus from different habitats, two parts of substitute Radix Astragali, three parts of alfalfa and one part of hollyhock were collected. Total DNA was extracted from all the samples and amplified by PCR. The amplified products were sequenced The corresponding sequences were also downloaded from GenBank. The K-2-P distance between species was calculated using MEGA 4, and finally its ITS sequence was used to construct its phylogenetic tree. Results: The ITS sequences of 19 samples were obtained, which were 646-650 bp in Astragalus Mongolica, 646-650 bp in Astragalus membranaceus, 664 bp in Radix Astragali, 659 bp in alfalfa, and 728 bp in Hollyhock, In registration, access to registration number. Clustering analysis of phylogenetic trees using ITS sequences to reconstruct the phylogenetic tree clearly distinguished Astragalus from its adulterants. Conclusion: ITS sequence can successfully identify astragalus and its adulterated counterfeit products, which can be used as a molecular identification method of astragalus and its adulterants.