基于ITS2条形码的曼陀罗属药用植物DNA分子鉴定  

DNA Molecular Identification of Datura Medicinal Plants Using ITS2 Barcode Sequence

作　　者：吴亚男[1] 许亮[1] 陈靓[1] 王冰[1] 赵容[1] 

机构地区：[1]辽宁中医药大学,辽宁大连116600

出　　处：《中药材》2015年第9期1852-1857,共6页Journal of Chinese Medicinal Materials

基　　金：辽宁中医药大学杰出青年基金(20121228);辽宁省高等学校杰出青年学者成长计划(LJQ2014101);"我国水生;耐盐中药资源的合理利用研究"项目(201407002)

摘　　要：目的:利用ITS2序列对曼陀罗属药用植物进行分子鉴定。方法:对曼陀罗属及烟草属植物样本的ITS2序列进行PCR扩增和测序。为扩大研究范围,又从Gen Bank中下载了上述两属植物样本的ITS2序列。所有序列经Codon Code Aligner软件拼接,DNAMAN软件比对分析,并采用MEGA 5.10计算相关数据,基于K2P模型构建聚类树(NJ树)。从ITS2网站上获得所测样本及下载序列的ITS2二级结构信息,分析各样本间ITS2序列二级结构的差异。结果:两属样本被聚为三大支。烟草属单聚为一支;木本曼陀罗单聚为一支,与前人研究结果相同:木本曼陀罗Brugmansia可同曼陀罗属Datura分为两属;曼陀罗属的其他物种聚在一支,且各品种种内样本均各为一支,表现出单系性,与其他种可明显区分;且每两支之间的bootstrap支持率均在95%以上。结论:ITS2序列在近缘物种间的系统学研究、品种的鉴定方面具有较大的应用潜力。Objective： To identify Datura medicinal plants in Solanaceae using DNA barcode technique and ITS2 sequence. Methods： The second internal transcribed spacer（ ITS2） of Datura and Nicotiana medicinal plant samples was amplified by PCR and sequenced. To expand scope of the research topic,ITS2 sequences of related species were downloaded from Gen Bank. Sequence assembly and consensus sequence generation were performed by Codon Code Aligner. All the ITS2 sequences in the study were compared and analyzed using software DNAMAN. The related data analysis and processing were performed using software MEGA 5. 10 and the NJ tree was constructed. The ITS2 secondary structure was predicted using ITS2 web server,and distinguishing differences of the ITS2 secondary structures of the samples. Results： In the cluster dendrogram,all of samples were clustered into three branches. The plants of Nicotiana were clustered into a single branch,and the same as Datura（ Brugmansia） arborea,which is similar to the results of previous studies. These proved that Brugmansia and Datura belonged to two different genera. Other species of Datura were clustered into a branch where all the intra-species samples were clustered to one branch respectively and obviously distinguished,which showed monophyletic. Bootstrap support rates between each two branches were more than 95%. Conclusion： ITS2 sequences have great potential in terms of systematics study and variety identification.

关 键 词：ITS2 分子鉴定 曼陀罗属 二级结构 

分 类 号：R282.5[医药卫生—中药学]