市售覆盆子药材DNA条形码鉴定研究

Identification of Commericially Available Rubi Fructus Based on DNA Barcoding Technology

目的基于ITS2条形码序列检测市场销售覆盆子药材,为保证覆盆子药材使用的正确性和安全性提供一种新的鉴定手段。方法获取掌叶覆盆子及其5种常见同属易混种ITS2序列,以及GenBank上下载的共计48条序列。使用Gene Tool软件分析ITS2序列长度,GC含量和变异位点等情况,利用Clustal X和MEGA 7.0软件计算遗传距离和构建邻接系统发育聚类树。同时随机检测24份市售覆盆子药材,利用中药材DNA条形码鉴定系统和构建邻接系统发育聚类树确定物种,鉴别真伪。结果掌叶覆盆子基原植物可与其同属易混种进行明显区分;市售药材中正品22份,伪品1份,混合物1份。结论基于ITS2序列的DNA条形码技术能够成功鉴定市场销售的掌叶覆盆子及其混伪品。

OBJECTIVE To provide a new method for the identification of Rubi Fructus in medicial market based on ITS2 barcode, and guaranting the accuracy and safety of Rubi Fructus. METHODS Combined with the sequences downloaded from GenBank, a total of 48 ITS2 sequences of Rubus chingii Hu and its confusable species of genus Rubus were obtained. Sequence characteristics such as length, GC contents, and mutation loci were analyzed through Gene Tool software, while genetic distance calculation and build-up of neighbor-joining(NJ) phylogenetic tree were carried out by Clustal X and MEGA 7.0. Simultaneously, 24 samples of Rubi Fructus collected from markets randomly were detected by Chinese herbal medicine DNA barcoding identification system and build-up of NJ phylogenetic tree. RESULTS The results revealed that R. chingii Hu and its adulterants could be differentiated obviously. The 22 samples of Rubi Fructus collected from markets were genuine, 1 sample was counterfeit and 1 sample was mixture. CONCLUSION The DNA barcoding technology based on ITS2 sequences can identify R. chingii Hu from its adulterants successfully.