蒙药材草乌叶DNA条形码研究  

DNA barcodes of Aconiti Kusnezoffii Folium as Mongolian medicine

作　　者：李谦[1,2] 王维宁 过立农[2] 刘杰[2] 郑健[2] 马双成[2] 

机构地区：[1]中国药科大学,南京210009 [2]中国食品药品检定研究院,北京100050 [3]辽宁省药品检验检测院,沈阳110036

出　　处：《药物分析杂志》2016年第6期1044-1052,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：国家食药监总局药化注册司民族药专项;中药质量与安全标准研究创新团队

摘　　要：目的:建立草乌叶与其易混品的分子鉴定方法。方法:通过聚合酶链式反应(PCR)对草乌叶及9种混伪品的候选序列(ITS、ITS2、matK、rbcL、psbA-trnH)进行扩增,比较各序列的扩增效率和测序成功率,并进行种内、种间遗传变异分析和barcoding gap分析。结果:ITS和ITS2序列GC含量较高,种间变异明显大于种内变异,根据barcoding gap图显示种内、种间遗传距离重叠较小,适合物种的区分,基于K2P距离进行聚类分析,ITS序列呈现较好的单系性。结论:ITS序列可应用于蒙药材草乌叶与其混伪品的鉴定,为民族药鉴定提供新方法。Objective： To establish a molecular identification method for Aconiti Kusnezoffii Folium and its adulterants. Methods： The candidate sequences （ ITS, ITS2, matK, rbcL, psbA-trnH） of Aconiti Kusnezoffii Folium and nine adulterants were amplified with polymerase chain reaction （ PCR ）. The amplification efficiency and success rate of sequencing were compared. The inter-specific and intra-specific variations, as well as the barcoding gap was analyzed. Results： ITS and ITS2 sequences had higher GC content, and the inter-specific divergence was more obvious than intra-specific variation. The figure of barcoding gap showed that the overlap between inter-specific distances and intra-specific distances was smaller than other loci, which was more suitable for species identification. The NJ trees based on K2P distances showed that ITS sequences revealed better monophyly.Conclusion： ITS sequence can be used to differ Aconiti Kusnezoffii Folium from adulterants, and be employed as a new method for identification of ethnodrug.

关 键 词：民族药 蒙药 草乌叶 混伪品 基因检测 DNA条形码 分子鉴定 物种鉴定 基原鉴定 聚合酶链式反应(PCR) ITS序列 

分 类 号：R917[医药卫生—药物分析学]