北柴胡与银州柴胡的ITS条形码鉴定研究  

Molecular Identification of Bupleurum chinense DC. and B.yinchowense Shan et Y. Li Based on ITS Sequence

作　　者：袁伯川 马永生[1] 杨瑞[1] 周姗[1] 林瑞超[1] 刘颖[1] 

机构地区：[1]北京中医药大学中药学院中药品质评价北京市重点实验室,北京100102

出　　处：《生物技术通讯》2016年第1期101-105,共5页Letters in Biotechnology

基　　金：国家科技重大专项(2014ZX09304307001-017)

摘　　要：目的：建立基于内转录间隔区（ITS）序列的北柴胡与银州柴胡分子鉴定新方法。方法：以来自北京、宁夏、山西、甘肃、河北的10个居群共125份北柴胡及银州柴胡样品为实验材料,提取其总DNA,并PCR扩增其ITS序列,对序列进行双向测序,应用Clustal X及BLAST查错,用MAGA 5.0进行序列分析,计算种内、种间遗传距离,构建邻接树,并统计不同单倍型的分布情况。结果：北柴胡与银州柴胡的ITS序列全长为604 bp,有7个变异位点,共计6种单倍型,其中单倍型1为银州柴胡所特有,单倍型2~6为北柴胡所特有;ITS序列种内最大遗传距离0.005,种间最小遗传距离0.007。结论：利用ITS序列可快速准确地鉴定北柴胡与银州柴胡。Objective： To establish a new method for molecular identification of Bupleurum chinense DC. and B. yinchowense Shan et Y. Li. Methods： 125 samples from 10 populations were used as plant materials, total DNA of which were extracted and then the internal transcribed spacer（ITS） sequences were obtained by PCR. All of the ITS sequences were checked and aligned using Clustal X and BLAST. The genetic distances were calculated and Neighbor Joining tree were constructed using MEGA 5.0, and the distribution of different haplotypes were de- scribed. Results： 125 ITS sequences with a full length of 604 bp were obtained. There were 6 ITS haplotypes based on 7 variable sites. Among them, haplotype 1 was specific for B.yinchowense, and haplotype 2-6 were specif- ic for B.chinense. The largest intraspecific genetic distance was 0.005, while the smallest interspecific genetic dis- tance was 0.007. Conclusion： B.chinense and B.yinchowense can be identified by ITS sequence rapidly and effec- tively.

关 键 词：北柴胡 银州柴胡 内转录间隔区 分子鉴定 

分 类 号：Q945[生物学—植物学]