高良姜及其混淆品rDNA ITS序列的分析与鉴别

Analysis and Identification of rDNA ITS Sequences of Alpinia officinarum Hance and Its Adulterants

用聚合酶链式反应(PCR)直接测序法测定和分析不同产地野生和农家品种的高良姜及其3种混淆品(山姜、华山姜和大高良姜)的核糖体DNA内转录间隔区(rDNAITS)序列,为高良姜种质资源研究和真伪鉴别提供分子数据.ITS序列分析结果表明:高良姜种内序列同源性达100%,测序结果中发现杂合位点,且广西样品杂合位点的两个碱基所占的比例与其它地方样品的不同.高良姜及其混淆品经测序、比对、排序得到的812 bp序列中有61个变异位点,60个是信息位点,同源性达96.32%,其中ITS1和ITS2区中的11个位点在高良姜及其混淆品中差别明显,可以鉴别高良姜及其混淆品.同时发现,基于DNA序列的高良姜及其混淆品的系统分类结果与形态学的分类结果不完全一致,有待进一步研究探讨.

In order to provide molecular data for the germplasm resource study on Alpinia officinarum （A officina rum） Hance and to identify A. officinarum Hance from its adulterants, the rDNA internal transcribed spacer （ITS） sequences of A. officinarum Hance and its adulterants were determined and analyzed by means of polymerase chain reaction （PCR）. Wild and cultivated A. officinarum Hance collected in different areas, and three adulterants including A. japonica （Thunb.）, A. chinensis （ Retz. ） Rosc. and A. galanga （ L. ）, were used as samples. The results of sequence analysis indicate that （ 1 ） the intraspecies sequence homology of A. officinarum Hance is up to 100% ; （2） there exist heterotic sites in all samples of A. officinarum Hance, and the proportion of the two bases in heterotic sites in the sample from Guangxi is different from that in other samples; （3） there are 61 variable sites （including 60 informative sites） in the 812bp sequences among A. officinarum Hance and its adulterants, with a homology of 96.32% ; （4） 11 sites in the ITS1 and ITS2 regions can be used to identify A. officinarum Hance and its adulterants due to the great difference of the sites in different samples; and （5） some further researches should be performed because the classification results of A. officinarum Hance and its adulterants based on DNA sequences do not totally accord with those based on morphologic characteristics.