高良姜与混淆品rDNA ITS序列的分析与鉴别

用PCR直接测序法测定和分析不同产地野生和种植品种的高良姜(Alpinia officinarum Hance)，以及山姜（A. japonica（Thunb.） Miq.）、华山姜（A. chinensis (Retz.) Rosc.）和大高良姜(A. galanga (L.) Willd )等三种混淆品的rDNA ITS区序列，为高良姜种质资源研究和真伪鉴别提供分子数据。经测序、比对和排序得到812bp序列，包括18S3’端部分序列，ITS1、5.8S、ITS2全部序列和26S5’端部分序列。序列分析结果显示，高良姜种内序列高度一致，同源性达100%，测序结果中发现杂合位点，而广西样品杂合位点的两个碱基各占的比例与其它地方样品的不同，显示了高良姜种内rDNA ITS序列的细微差异。高良姜和混淆品的812bp序列中共有61个变异位点，60个是信息位点，同源性达96.32%，其中ITS1和ITS2中的11个位点在高良姜和混淆品中差别明显，可以鉴别高良姜和混淆品，因此rDNA ITS序列是高良姜真伪辨别的有效标记。基于DNA序列的高良姜和混淆品的系统分类结果与形态学的分类结果不完全一致，有待进一步的研究探讨。

Authanticaton of Alpinia officinarum Hance and adulterants by analysis of rDNA ITS sequences

In order to provided molecular data for germplasm diversity study of Alpinia officinarum Hance, and authentication between A. officinarum Hance and adulterants, rDNA ITS sequences were detected by PCR direct sequencing. The samples contented wild species and cultivars of Alpinia officinarum Hance from different location and adulterants of them, including A. japonica (Thunb.), (A. chinensis (Retz.) Rosc. And A. galanga (L.) Willd. 812bp sequences were obtained after sequencing and aligning, including partial sequences of 18S, complete sequences of ITS1 and 5.8S and ITS2, partial sequences of 26S. The sequences analysis indicated that the sequences of A. officinarum Hance intraspecies were of high uniformity, and the homology was 100%. However, heterotic sites were discovered in the sequencing results, and the bases proportion of heterotic sites of sample from Guangxi was different to the other samples, which revealed the tiny intraspecies variation of ITS sequences in A. officinarum Hance. There are 61 variable sites in the 812bp sequences among Alpinia officinarum Hance and adulterants, and 60 are informative sites, and their homology was 96.32%. 11 sites in ITS1 and ITS2 were obviously different between A. officinarum Hance and adulterants, and these sites can be used to authenticate them, therefore, ITS sequences are valuable maker for authenticating real and false of A. officinarum Hance. The classification results of A. officinarum Hance and adulterants based on DNA sequences are incomplete accordant to that based on morphologic characters, and further researches need to perform.