基于ITS2条形码对铁皮石斛及其混伪品分子鉴定的初步研究  

Molecular Identification of Dendrobium Officinale and its Adulterants Based on ITS2 Barcode

作　　者：刘依丽 冯尚国[1] 何仁锋[1] 赵妙玉 吴慧[1] 崔艺庆 陈倩[1] 王慧中[1] 

机构地区：[1]杭州师范大学生命与环境科学学院,浙江省药用植物种质改良与质量控制技术重点实验室,浙江杭州310036

出　　处：《杭州师范大学学报（自然科学版）》2014年第1期35-41,共7页Journal of Hangzhou Normal University(Natural Science Edition)

基　　金：国家自然科学基金项目(31070298;30870180);浙江省科技计划项目(2008C12081);浙江省自然科学基金项目(LQ13H280006);国家级大学生创新创业训练计划项目(201210346002)

摘　　要：通过分析铁皮石斛及其常见混伪品的ITS2序列差异,建立铁皮石斛药材真伪的快速鉴别方法.运用CodonCode Aligner V3.0软件对测序峰图进行校对拼接,基于隐马尔可夫模型的注释方法获得ITS2序列,采用MEGA5.0软件进行序列比对,种内种间距离计算及UPGMA聚类树构建,应用Koetschan等建立的ITS2数据库及网站预测ITS2二级结构.结果表明,铁皮石斛及其混淆品ITS2之间存在明显差异,所有铁皮石斛样品在UPGMA树上单独聚为一类.ITS2序列可以作为DNA条形码用于铁皮石斛与其混伪品的快速分子鉴别.In order to establish a fast identification method for Dendrobium of ficinale and its adulterants ,ITS2 sequence differences were analyzed .The ITS2 sequences were assembled using the CodonCode Aligner V3 .0 and was annotated with the HMM .Sequence alignment and genetic distances were computed using MEGA 5 .0 was used to compare the sequences , calculate the intraspecific and inter-specific distances and construct the clustering tree UPGMA . The ITS2 secondary structure was predicted using the ITS2 database and website which were built by Koetschan et al .A significant difference was observed between the ITS2 sequences of Dendrobium of ficinale and its adulterants ,namely ,all Dendrobium of ficinale samples singly clustered as a class in UPGMA tree .The study shows that ITS2 sequence can serve as the DNA barcode to identify Dendrobium of ficinale and its adulterants .

关 键 词：铁皮石斛 DNA条形码 ITS2 分子鉴定 ITS2 

分 类 号：R282.7[医药卫生—中药学]