摘要
为了对中药材甘草及其混伪品进行分子鉴定,以区分正品甘草与混伪品。从兰州市不同商家收集9种甘草,提取DNA,扩增ITS2序列并进行双向测序,在GenBank数据库收集甘草混伪品苦豆子、苦参、黄芪和山豆根的序列,应用MEGA 7.0进行序列变异分析,以Kimura 2-Parameter(K2P)模型计算遗传距离,并构建邻近(NJ)树。基于ITS2序列的序列变异、最近距离法和系统邻近树分析结果表明,该方法可将2015版《中华人民共和国药典》中规定的正品甘草与其混伪品进行区分。说明ITS2序列可有效区分5种甘草混伪品,为中兽医和中医临床应用提供保障依据。
Radix Glycyrrhizae is the most commonly used TCM or TCVM clinic,but the phenomenon of Radix Glycyrrhizae confounding products had caused great confusion to the clinical safety of TCM and TCVM,and it was difficult to satisfy the clinical drug use only by experience identification. In the study,molecular identification of Radix Glycyrrhizae and its adulteration was carried out to distinguish genuine Radix Glycyrrhizae from adulterates. In this experiment,9 kinds of Radix Glycyrrhizae were collected from different merchants in Lanzhou,ITS2 sequence was amplified and bidirectional sequencing was carried out after DNA was extracted,and the results were provided by sequencing company. Radix Glycyrrhizae,Radix Sophorae Flavescentis,Radix Astragali and Radix Sophorae Tonkinensis,which were mixed with Radix Glycyrrhizae. The sequence source of Radix Glycyrrhizae adulterant were collected from GenBank. The sequence of astragalus root and soybean root was analyzed by MEGA7.0. The genetic distance was calculated by Kimura 2-Parameterus K2 P model,and the adjacent tree was constructed. Based on the sequence variation of ITS2 sequence,the results of nearest distance method and phylogenetic proximity tree analysis showed that licorice,as a product of Chinese Pharmacopoeia,could be distinguished from its adulterates. 5 kinds of licorice adulterants can be distinguished effectively by the sequence of ITS2,which can provide evidence for traditional Chinese veterinary medicine and TCM clinic.
作者
白东东
李新圃
杨峰
罗金印
张行
李宏胜
BAI Dongdong;LI Xinpu;YANG Feng;LUO Jinyin;ZHANG Hang;LI Hongsheng(Key Laboratory of Veterinary Pharmaceutical Discovery, Ministry of Agriculture, Key Laboratory of New Animal Drug Project of Gansu Province,Lanzhou Institute of Animal Husbandry and Pharmaceutical Sciences,Chinese Academy of Agricultural Sciences, Lanzhou Gansu 730050, China)
出处
《中兽医医药杂志》
2019年第3期14-16,共3页
Journal of Traditional Chinese Veterinary Medicine
基金
国家重点研发项目"畜禽群发普通病防控技术研究"(2017YFD0502200)
甘肃省国际科技合作项目(17YF1WA169)
中国农业科学院奶牛疾病研究创新工程项目
关键词
甘草
ITS2序列
分子鉴定
混伪品
Radix Glycyrrhizae
ITS2 sequence
molecular identification
adulteration
