摘要
为验证DNA条形码序列在物种鉴定方面及物种的系统发育方面的广适性,本研究以黎药植物木麻黄为研究对象,收集木麻黄及其近缘种和混伪品22份样本,采用4对引物(ITS,ITS2,trnH-psbA,rbcL)分别进行PCR扩增,产物进行双向测序。利用CodonCode Aligner软件进行序列的拼接,采用MEGA5.1进行数据的处理。结果表明,利用ITS/ITS2可以区分木麻黄与其近缘种以及混伪品,样品的psbA-trnH,rbcL和ITS/ITS2 4个序列中,ITS/ITS2序列具有明显的DNA gap。基于ITS/ITS2序列构建的NJ树结果表明,木麻黄属所有样品聚为一类,呈现明显的单系性,与其亲缘关系较近的是藤黄科。因此,ITS/ITS2序列可以准确鉴定黎药的基原植物。并能从分子水平揭示木麻黄的系统演化规律。
In this research,Casuarina eguisetifolia Linn was used to verify the broadly suitability of DNA barcodes in identification of Li-medicine plants and systematic development of species.The genomic DNA of 22 samples collected C.eguisetifolia and its adulterants were amplified by 4 pairs of primers respectively(ITS(internal transcribed spacer),ITS2(internal transcribed spacer 2),trnH-psbA,rbcL) and sequenced bi-directionally.Obtained sequences were assembled using CodonCode Aligner.The dates were analysised using MEGA5.1 in ac cordance with the kimura 2-parameter(K2P) model.The neighbor-joining(NJ) phylogenetic trees were construct ed.Our study demonstrated the efficacy of ITS/ITS2 to distinguish between C.eguisetifolia and other adulterants species at the molecular level.Comparative to the primer of trnH-psbA and rbcL,there was a obviously DNA gap.The NJ trees showed that the several species of Casuarina can be classified to same types to show a obviously monophyly,which the nearest family was Guttiferae.Therefore,ITS/ITS2 regions can accurately distinguish the original plant of Li-medicine.The systematic evolution of Casuarina can be verified in the molecular level.
出处
《世界科学技术-中医药现代化》
北大核心
2013年第3期397-403,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
海口市科技工业信息局重点科技项目(2011-0036):黎药的DNA条形码研究
负责人:唐历波
关键词
木麻黄
ITS
ITS2
物种鉴定
系统发育
Casuarina equisetifolia
ITS/ITS2
species identification
systematic evolution