基于ITS2序列对市售白扁豆的鉴别研究

Identification of Commercial Baibiandou(Dolichos Lablab)Based on ITS2 Sequence

目的了解不同来源市售白扁豆核糖体内部转录间隔区2(internal transcribed spacer 2,ITS2)序列信息,并基于该序列对其进行鉴定。方法收集云南、安徽、广东等地以及进口白扁豆31份,采用试剂盒法提取DNA,聚合酶链式反应(PCR)扩增ITS2序列后并测序,对峰图校对和拼接,与从GeneBank数据库下载的71条白扁豆及其混伪品ITS2序列进行综合分析,并构建基于邻接法的系统发育树,预测ITS2二级结构。结果31份市售白扁豆均测序成功,白扁豆ITS2条形码序列存在两个变异位点,基于ITS2条形码序列及其二级结构均可将白扁豆与其近缘混伪品区分。结论不同产地和来源白扁豆ITS2条形码序列无明显差异,ITS2条形码序列可以用于白扁豆的真伪鉴别。

Objective To understand the sequence information of internal transcribed spacer 2(ITS2)in baibiandou ribosomes from different sources and identify them based on this sequence.Methods 31 samples of baibiandou from Yunnan,Anhui,Guangdong and other places as well as imported baibiandou were collected.DNA was extracted by the kit method.The ITS2 sequence was amplified by PCR and then sequenced.The peak map was proofread and spliced.A comprehensive analysis was conducted with 71 baibiandou and their adulterants ITS2 sequences downloaded from GeneBank.A phylogenetic tree based on the NJ method was constructed and the secondary structure of ITS2 were predict.Results 31 commercially available baibiandou were sequenced successfully.There were two variation sites in the ITS2 barcode sequence of baibiandou.The ITS2 barcode sequence and its secondary structure can be used to distinguish baibiandou from its closely related varieties.Conclusion There is no significant difference in the ITS2 barcode sequence of baibiandou from different habitats and sources.The ITS2 barcode sequence can be used to identify the true and false baibiandou.