东北当归属植物DNA条形码分子鉴定研究

Molecular identification of plants in Genus Angelica in Northeast of China based on DNA barcoding

目的通过分析6种当归属植物的IST2,matK rbcL和trnH-psbA条形码序列,探讨东北当归属植物鉴定新方法。方法共收集18份样品,以ITS2,matK,rbcL和trnH-psbA作为条形码序列,对当归属植物提取基因组DNA,PCR扩增并进行双向测序。所得序列经CodonCode Aligner拼接并剪切后,采用MEGAX软件进行序列对比分析,计算遗传距离,分析比较种内、种间序列差异,并构建系统邻接树。结果与其他DNA条形码候选序列相比,ITS2序列在所研究的东北当归属植物中的扩增和测序效率均为100%,其种内及种间变异、barcoding gap均具有明显优势,其鉴定成功率达到100%。结论ITS2序列作为DNA条形码可较好地用于东北当归属植物DNA条形码的鉴定研究。

Objective To explore a new identification method for the plants in Genus Angelica in Northeast of China by analyzing IST2,matK,rbcL and trnH-psbA barcode sequences of 6 plants in Genus Angelica.Methods The ITS2,matK,rbcL and trnH-psbA of ribosomal DNA of 18 medicinal materials of Angelica was amplified and sequenced by bi-directional sequencing of PCR products.Sequence assembly and consensus sequence generation were performed by using CodonCode Aligner.Phylogenetic study was performed using software MEGAX.Genetic distances were calculated and analyzed and the phylogenetic tree was constructed.Results Compared with other candidate DNA barcode sequences,the ITS2 sequence has 100%amplification and sequencing efficiency in the studied plants,its intra-and inter-species variation and barcoding gap have obvious advantages,and the identification success rate of ITS2 sequence in Genus Angelica in Northeast of China reached 100%.Conclusion ITS2 sequence as DNA barcoding technique can be used to identify the plants in Genus Angelica in Northeast of China.