目的:探讨基于ITS2条形码序列鉴定高良姜及其混淆品的新方法。方法:本研究对高良姜及其混淆品6个物种9份样品的ITS2序列进行PCR扩增和测序,同时扩大研究范围从Gen-Bank上下载同属共15个物种37个样本。用MEGA4.1计算其种间、种内的K2P距离,并分析各样本间ITS2序列二级结构的差异,最后利用ITS2序列重构其系统发育树。结果:高良姜基源植物种内最大K2P距离为0.0171,与混伪品的种间最小K2P距离为0.0469;高良姜及其混淆品的ITS2二级结构存在明显差异;重构的系统发育树显示高良姜的不同来源聚在一支,能较好与混淆品区分。结论:ITS2序列能够成功鉴定高良姜及其易混淆品,可以作为高良姜及其混淆品的分子鉴定工具。

This study aimed to discriminate Alpinia officinarum and its adulterants using ITS2 sequence.DNA samples were extracted from Alpiniae specimens.The ITS2 sequence was amplified by PCR and sequenced bidirectionally.Sequences were assembled by CodonCode Aligner.The intra-specific and inter-specific K2P distances of Alpinia officinarum and its adulterants were calculated.And the N-J tree was constructed by MEGA V 4.1.The ITS2 secondary structure was predicted.The results showed that the maximum intra-specific K2P distance was 0.0171,while the minimum inter-specific K2P distance was 0.0469.Different samples of Alpinia officinarum were clustered.And the N-J tree was able to distinguish Alpinia officinarum from its adulterants.The Alpinia officinarum was obviously differentiated from its adulterants by the ITS2 secondary structure.It was concluded that ITS2 sequence was able to identify Alpinia officinarum and its adulterants.It was an efficient molecular identification method for Alpinia officinarum and its adulterants.

北京市科技计划项目(D08080203640901):羌活、石斛药材质量评价和分子生物学鉴定示范研究,负责人:陈士林