摘要
目的:研究吴茱萸药材3种不同基原植物及伪品楝叶吴茱萸的亲缘关系和真伪鉴别。方法:采用DNA条形码技术,对ITS2区段进行PCR扩增和测序,使用MEGA 5.1软件计算种内、种间Kimura-2-parameter遗传距离,采用邻接(NJ)法构建其系统发育树。使用DNAstar 5.0软件比对特异性位点。结果:4个物种共18个样本的ITS2序列长度均为220bp,产生了17个单核苷酸多态性位点,其中特异性位点7个;正品吴茱萸药材的3种基原植物种内最大遗传距离为0.023,种间遗传距离为0.036-0.039,与楝叶吴茱萸的最小种间遗传距离为0.042;NJ系统发育树表明:基于ITS2序列聚类的吴茱萸基原植物可与楝叶吴茱萸有效区分,3种基原植物同一品种样本优先聚类,其次是同一产区聚类。结论:ITS2 DNA条形码可以准确有效地用于吴茱萸属植物亲缘关系研究和真伪品的鉴别。
Objective： To study the genetic relationship and molecular identifcation of three kinds of Evodia rutaecarpa original plants and one kind of adulterant plant. Methods： DNA barcoding technology was used to amplify the second internal transcribed space （ITS2） section and the sequences were sequenced. The distances of intraspecies and interspecies, were calculated by using the software of MEGA 5.1 with Kimura-2-parameter （K2-P） model and the phylogenetic tree was constructed by using the neighbor-joining （NJ） method. Specific sites of these sequences were aligned by using the software of DNAstar 5.0. Results： The results showed that the length in ITS2 region of four species were same in all eighteen samples, which were 220bp. 17 single- nucleotide polymorphism sites were produced in 220 bp, in which seven specific sites. The longest intraspecific genetic distance of Evodia rutaecarpa was 0.023 and their interspecific genetic distance ranged from 0.036 to 0.039. Their shortest interspecific genetic distance with the adulterant was 0.042. The phylogenetic tree showed that ITS2 could effectively distinguish Evodia rutaecarpa from adulterant. The plants from the same species were clustered first, followed by the same areas clustered. Conclusion： DNA barcoding of ITS2 can distinguish accurately and efficiently genetic relationship of Evodia plants and control the quality.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2013年第12期3681-3684,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
"十二五"国家科技支撑计划项目(No.2011BAI04B04)
江西中医药大学重点学科培养计划项目(No.2012jzzdxk048)~~
关键词
吴茱萸
ITS2序列
亲缘关系
分子鉴别
Evodia rutaecarpa
ITS2 sequences
Genetic relationship
Molecular identification
