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.
出处
《药物分析杂志》
CAS
CSCD
北大核心
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
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
ethnodrug
Mongulian medicine
Aconiti Kusnezoffii Folium
adulterants
genetic testing
DNAbarcodes
molecular identification
species identification
origin identification
polymerase chain reaction （ PCR ）
internal transcribed spacer of nuclear ribosomal DNA
