基于trnL-trnF序列分析的何首乌PCR-RFLP分子鉴别  

Molecular Authentication of Fallopia multiflora by PCR-RFLP Based on the trnL-trnF Analysis

作　　者：郑传进[1] 生书晶[2] 赵树进[2] 

机构地区：[1]广东药学院食品科学学院,广东中山528458 [2]广州军区广州总医院,广东广州510010

出　　处：《中药材》2012年第4期543-547,共5页Journal of Chinese Medicinal Materials

基　　金：广东省自然科学基金(06019716)

摘　　要：目的:建立何首乌(Fallopia multiflora)DNA分子鉴别技术。方法:对何首乌与其近缘种及其混淆品的trnL-trnF(trnL基因和trnF基因间隔区)序列进行比较分析。结果:何首乌与其近缘种及其混淆品的trnL-trnF差异率为2.1%～22%,何首乌种内各居群间trnL-trnF差异率为0%～1.5%。基于何首乌与其近缘种及其混淆品的trnL-trnF序列的差异,找出一个位于trnL5'-trnL3'间区的何首乌特征性Xba I酶切位点(T↓CTAGA),用Xba I酶对不同采集地的何首乌样品trnL-trnF序列扩增产物酶切后均得到含约804～819 bp和256 bp两个片段的PCR-RFLP图谱,而其混淆品trnL-trnF序列扩增产物因不能被Xba I酶切,图谱呈单一条带。结论:利用建立的PCR-RFLP方法可以很好地区分何首乌及其混淆品植物。Objective：To establish a method for the molecular authentication of Fallopia multiflora. Methods： The trnL-trnF re- gions of Fallopia multiflora and its closely related species and/or adulterants were sequenced and analyzed. Results ： It was found that the trnL-trnF sequence divergences between Fallopia multiflora and its closely related species and/or aduherants were 2. 1% -22%. While the intra-species trnL-trnF divergences of FaUopia multiflora were 0% - 1.5%. Based on the trnL-trnF regional variations, an endonuclease Xba I （T ,[ CTAGA） restriction site specific to Fallopia multiflora was detected. The Fallopia multiflora trnL-F polymer- ase chain reaction product could be cleaved by Xba I into two pieces, 804 -819 bp and 256 bp each, whereas the restriction endonucle- ase could not digest the trnL-trnF polymerase chain reaction product of its closely related species or adulterants. The restriction patterns analyzed for restriction enzyme Xba I were found to be identical in all Fallopia multiflora individuals from different geographical regions in China. Conclusion： The assay based on polymerase chain reaction amplification of the trnL-trnF fragment of chloroplast DNA and subsequent restriction fragment length polymorphism can be used as a general test to identify Fallopia multiflora.

关 键 词：何首乌 分子鉴别 trnL基因和trnF基因间隔区 PCR.RFLP 

分 类 号：R282.5[医药卫生—中药学]