基于psbA-trnH序列的芦根及其混伪品DNA条形码鉴定  

Identification of Phragmitis Radix and Its Adulterants Based on psb A-trnH Sequence by DNA Barcoding Method

作　　者：韩正洲 吴正军 魏伟锋 陈新连 林余霖[2] 王瑀[2] HAN Zheng-zhou;WU Zheng-jun;WEI Wei-feng;CHEN Xin-lian;LIN Yu-lin;WANG YU(Sanfiu Medical ＆ Pharmaceutical Co.,Ltd,Shenzhen 518110,China;Key Lab of Chinese Medicine Resources Conservation,State Administration of Traditional Chinese Medicine of the People＇s Republic of China,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences ＆ Peking Union Medical College,Belting 100193,China;NERC for the Pharmaceutics of Traditional Chinese Medicines,Benxi 117004,China)

机构地区：[1]华润三九医药股份有限公司,广东深圳518110 [2]中国医学科学院,北京协和医学院,药用植物研究所,国家中医药管理局中药资源保护重点研究室,北京100193 [3]国家中成药工程技术研究中心,辽宁本溪117004

出　　处：《中国现代中药》2018年第10期1225-1229,1241,共6页Modern Chinese Medicine

基　　金：深圳中药现代化与新药创制工程实验室提升项目(深发改[2015] 1962号)

摘　　要：目的:应用psb A-trnH序列对芦根及其混伪品进行鉴定研究,为该药材的临床用药安全和市场监管等方面提供参考。方法:对芦根及其基原物种进行DNA提取、PCR扩增psb A-trnH序列、双向测序,结合Gen Bank下载序列,对序列进行比对、计算遗传距离、构建NJ系统聚类树。结果:芦根psb A-trnH序列长度为482～483 bp,种内有2个变异位点,有2种单倍型,G+C含量为37. 7%～37. 8%。芦根psb A-trnH序列种内遗传距离为0～0. 004 2,芦根与其混伪品的种间遗传距离为0. 008 3～0. 036 1,大于芦根种内最大遗传距离。基于psb A-trnH序列构建的NJ聚类树显示,芦根样品单独聚为一支,其混伪品各自聚为一支。结论:应用psb A-trnH序列可以鉴别芦根及其混伪品。Objective： This research aims to identify Phragmitis Rhizoma and its adulterants using psbA-trnH sequence. It will provide the foundation for its clinical medication and accurate market standardized management. Methods： Genomic DNA of Phragmitis Rhizoma and its botanical origins were extracted. Then the psbA-trnH regions were obtained by PCR amplification mid sequenced bidirectionally. The psbA-trnH sequences were aligned with the sequences from GenBank. The genetic distances were com- puted using MEGA 6. 0 in accordance with the Kimura 2-parameter（K2P） model and Neighbor-Joining（NJ） tree was constructed with 1000 bootstrap. Results： The psbA-trnH sequence length of Phragmites commanis was 482-483 bp and the G ＋ C content was 37. 7% - 37. 8%. There were two variable sites and one insert/delete site in P. corrormnis. The intra-specific divergence of P. communis was 0-0. 004 2. The inter-specific divergence between Phragmitis Rhizoma and its adulterants was 0. 008 3-0. 036 1, which was larger than the maximum intra-specific divergence of P. communis. The NJ tree showed that P. comnmnis and its adulterants were divided into two groups obviously based on the psbA-tmH sequences. Conclusion： The results indicated that the psbA-trnH sequence could be a valua- ble marker to identify Phragmitis Rhizoma and its adulterants.

关 键 词：芦根 鉴定 PSB A-trnH序列 DNA条形码 混伪品 

分 类 号：R282.5[医药卫生—中药学]