基于COI条形码序列的《中国药典》动物药材鉴定研究

Application of COI-based DNA Barcoding for Identifying Animal Medical Materials in the Chinese Pharmacopoeia

药用动物是我国中药资源的重要组成部分,线粒体COI基因被公认为动物界中标准的DNA条形码基因,但利用此技术系统研究药用动物或动物药材鉴定的报道不多。本研究选取药典中45个动物药材,涉及基原物种51种,分析样品COI序列的种内种间变异情况,barcoding gap,鉴定效率等,考察COI序列鉴定动物药材的有效性和准确性。结果表明,种间变异最小值远大于种内变异最大值,barcoding gap图显示种间变异和种内变异重合较少,鉴定效率较高,除节肢动物门外,在物种水平和属水平上,鉴定效率均为100%,所构建的动物药材及其混伪品的NJ树能很好地区分正品来源与其混伪品。因此COI序列作为DNA条形码适用于药典中动物药材的鉴定。本研究为COI条形码准确鉴定药典中动物药材提供了分子依据,扩充了DNA条形码数据库中药用动物序列,并建立了利用此技术鉴定动物药材的具体技术流程。

Medicinal animals are important part of Traditional Chinese medicine resources in China.Cytochrome c oxidase subunit I（COI） was selected as the standard DNA barcoding sequence for animal medical materials.In this study,the 51 animal species from 45 animal medical materials in the Chinese Pharmacopoeia were selected and the intra-specific variation and the inter-specific divergence,the barcoding gap,the identification efficiency of their COI sequences were analyzed.The results showed that the inter-specific divergence is higher than intraspecific distance.The barcoding gap existed between inter-specific sequence divergence and intra-specific distance.The identification efficiencies were 100% both at the genus and species level except the Arthropoda.The cluster dendrogram exhibited that different species distinguished from others.Therefore,COI sequence as a barcode is suitable to identify the species of animal medical materials in Chinese Pharmacopoeia.