摘要
目的:通过分析竹叶花椒及其近缘种药材的核糖体DNA内转录间隔区2(ITS2)条形码序列,探讨DNA条形码技术鉴别竹叶花椒及其近缘种药材的可行性。方法:对样品进行DNA提取、扩增、测序,并从GenBank数据库下载ITS2序列,共获得竹叶花椒及其近缘种药材ITS2序列43条,基于Basic Local Alignment Search Tool(BLAST)法、临接法(NJ)、Kimura 2-parameter(K2P)遗传距离和ITS2序列二级结构进行鉴定分析。结果:BLAST结果与性状鉴定结果一致;竹叶花椒种内平均K2P遗传距离小于其与花椒、青花椒、野花椒的种间平均K2P遗传距离;NJ系统聚类树和ITS2二级结构可直观地区分竹叶花椒及其近缘种。结论:应用ITS2条形码可以有效地鉴别竹叶花椒及其近缘种药材。
Objective:To explore the feasibility of using DNA barcoding technology to identify Zanthoxylum armatum and its closely related species,the internal transcribed spacer 2(ITS2)barcoding sequences were analyzed.Methods:By DNA extraction,amplification and sequencing of experimental samples,and downloading the ITS2 sequences from GenBank,a total of 43 ITS2 sequences of Z.armatum and its closely related species were obtained.Identification was conducted based on BLAST method,Neighbor-Joining(NJ)phylogenetic tree method,Kimura 2-parameter(K2P)genetic distance method,and secondary structure of ITS2 sequence.Results:The results of BLAST were consistent with those of morphological identification.The average intraspecific K2P genetic distance of Z.armatum was smaller than the average interspecific K2P genetic distance between it and Z.bungeanum,and Z.schinifolium,and Z.simulans.Both NJ phylogenetic tree and ITS2 secondary structure can distinguish Z.armatum and its closely related species intuitively.Conclusion:The ITS2 barcoding method can be used to identify Z.armatum and its closely related species.
作者
齐景梁
高必兴
苟琰
耿昭
卿艳
周娟
姜卫东
QI Jing-liang;GAO Bi-xing;GOU Yan;GENG Zhao;QING Yan;ZHOU Juan;JIANG Wei-dong(Sichuan Institute for Drug Control, Key Laboratory of Quality Evaluation of Chinese Patent Medicines, NMPA, Chengdu ChinaChengdu University of Traditional Chinese Medicine, Chengdu China)
出处
《中国现代中药》
CAS
2021年第5期786-791,814,共7页
Modern Chinese Medicine
基金
四川省中医药管理局科学技术研究专项(2018YY002)。
关键词
竹叶花椒
内转录间隔区2
DNA条形码
中药鉴定
Zanthoxylum armatum DC.
ITS2
DNA barcoding
authentication of Chinese medicine
