蒙药材草乌叶DNA条形码研究

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.