基于ITS序列鉴别特色民族药材土牛膝及其混伪品的研究  

DNA Molecular Identification of Achyranthes Aspera and Its Adulterants Based on ITS Barcode

作　　者：胡亮 方磊 李瑞莲 王湘波 周明 HU Liang;FANG Lei;LI Ruilian;WANG Xiangbo;ZHOU Ming(Hunan Institute for Drug Control,Hunan Pharmaceutical Excipients Testing and Inspection Center,Hunan Engineering&Technology Research Center for Pharmaceutical Quality Evaluation,Changsha Hunan 410001,China)

机构地区：[1]湖南省药品检验研究院,湖南药用辅料检验检测中心,湖南省药品质量评价工程技术研究中心,湖南长沙410001

出　　处：《药品评价》2021年第3期141-143,共3页Drug Evaluation

基　　金：湖南省自然科学基金项目(2020JJ9036)。

摘　　要：目的:利用DNA条形码对特色民族药材土牛膝(粗毛牛膝、野生牛膝和柳叶牛膝)及其混伪品进行分子鉴定。方法:利用DNA条形码方法,分别对土牛膝及其混伪品的ITS和MatK基因片段进行扩增并双向测序,使用Codon Code Aligner软件对扩增序列进行拼接,用MEGA软件对数据比对分析,并基于K2P模型进行遗传距离分析,构建邻接(NJ)系统聚类树鉴定分析。结果:3种植物基原土牛膝药材粗毛牛膝、野生牛膝和柳叶牛膝的ITS序列种内最大K2P距离均小于其种间最小K2P距离;所构建的系统NJ聚类树图显示土牛膝药材与其混伪品可明显区分开,表现出良好的单系性;而基于MatK序列构建的系统聚类树图不能将不同基原土牛膝区分。结论:ITS作为DNA条形码可方便快捷地鉴别土牛膝及其混淆品。Objective:DNA barcodes were used to identify the Achyranthes aspera(Achyranthes Aspera L.,Achyranthes bidentata Blume and Achyranthes longifolia Makino)and its adulterants.Methods:The ITS nuclear gene fragment and MatK chloroplast gene fragment from Achyranthes aspera its adulterants were amplified and bidirectionally sequenced by DNA barcoding technology.The sequence was spliced by codoncode aligner,the data were compared and analyzed with mega software,and the genetic distance was analyzed based on K2P model,and the adjacency(NJ)clustering tree was constructed for identification analysis.Results:The ITS sequences of Achyranthes aspera,Achyranthes bidentata,and Achyranthes longifolia showed that the maximum intraspecific K2P distance was less than the minimum interspecific K2P distance.The NJ clustering tree showed that the radix Achyranthes aspera and its adulterants could be clearly distinguished,showing good monophyly;however,the cluster tree based on MatK sequence can not distinguish Achyranthes aspera.Conclusion:ITS can be used as DNA barcode to identify Achyranthes aspera and its adulterants.

关 键 词：土牛膝 DNA条形码 ITS 物种鉴别 

分 类 号：R282.5[医药卫生—中药学]