白前及其混伪品的ITS2分子鉴定  

Molecular Identification of Cynanchum stauntonii and Its Adulterants Based on DNA Barcoding(ITS2)

作　　者：林好[1] 陈镜安 李斯璐 豆蒙蒙 刘镛 LIN Hao;CHEN Jing-an;LI Si-lu;DOU Meng-meng;LIU Yong(School of Chinese Meteria Medica, Guangzhou University of Chinese Medicine, Guangzhou 510006, China;School of Pharmacy, Guangdong Medical University, Dongguan 523808, China;School of Basic Medicine, Guangdong Medical University, Dongguan 523808, China;Guangdong Key Laboratory for Research and Development of Natural Drugs, Guangdong Medical University, Zhan- jiang 524023, China)

机构地区：[1]广州中医药大学中药学院,广东广州510006 [2]广东医科大学药学院,广东东莞523808 [3]广东医科大学基础医学院,广东东莞523808 [4]广东医科大学广东天然药物研究与开发重点实验室,广东湛江524023

出　　处：《中药材》2017年第11期2531-2536,共6页Journal of Chinese Medicinal Materials

基　　金：广东省中医药局科研项目(20151264;20161139);广东医学院面上培育项目(M2014011;M2015015;M2016002);广东医学院大学生创新实验项目(2015ZZDM004);国家级大学生创新创业训练计划项目(201510571020;201610571002);省级大学生创新创业训练计划立项项目(201510571046;201510571079)

摘　　要：目的:采用ITS2序列鉴定白前及其易混伪品,为准确鉴别白前和保证临床用药的安全性提供参考。方法:白前与其混伪品的ITS2序列从Gen Bank数据库中获取,然后使用MEGA 6.06软件进行种内、种间序列变异分析、构建邻接系统树(NJ树)并计算遗传距离,运用ITS2数据库相似性搜索引擎预测ITS2二级结构。结果:白前ITS2序列长度在246~270 bp范围内,种内变异少且种间存在明显差异,与所有混伪品种间遗传距离为0.026~0.309。NJ树显示白前独聚一支,另外白前有明显ITS2二级结构特征,可与其混伪品明显区分。结论:ITS2序列能够有效地区分白前与其混伪品,为白前的用药安全提供技术保障。Objective： To discriminate Cynanchum stauntonii and its adulterants using internal transcribed spacer 2（ITS2） sequences,in order to identify Cynanchum stauntonii accurately and ensure clinical safety of this traditional Chinese medicine. Methods：The data of ITS2 regions of Cynanchum stauntonii and its adulterants were obtained from Gen Bank. In this study,MEGA 6. 06 software was used to analyze the interspecies and intraspecies sequence variation,construct the NJ（neighbor-joining） phylogenetic tree based on ITS2 sequences and calculate the genetic distance. The ITS2 secondary structure was predicted by similarity using the ITS2 database and website. Results： The length of ITS2 sequence was in the range of 246 ~ 270 bp,and there were significant differences between interspecies and intraspecies. The interspecific genetic distance between Cynanchum stauntonii and its adulterants ranged among 0. 026 ~0. 309. NJ tree showed Cynanchum stauntonii formed a individual cluster. On the other hand,Cynanchum stauntonii had its obvious ITS2 secondary structure which could be clearly distinguished with its adulterants.Conclusion： ITS2 can be used to correctly identify Cynanchum stauntonii and its adulterants,and for the rapid identification of Cynanchum stauntonii,so it can provide technical support for drug safety of Cynanchum stauntonii.

关 键 词：白前 DNA条形码 混伪品 ITS2 分子鉴定 

分 类 号：R282.5[医药卫生—中药学]