茜草“隐性品种”的鉴定及其质量的影响研究  

Study of ‘recessive varieties' of Rubiae radix et rhizoma on identification and its quality influence

作　　者：宋晓娜[1] 臧艺玫[1] 刘春生[1] 顾选[1] 马恺悦 马长华[1] 

机构地区：[1]北京中医药大学中药学院,北京100102

出　　处：《中华中医药杂志》2015年第7期2500-2503,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：高等学校博士学科点专项科研基金(No.20110013110006)~~

摘　　要：目的:采用DNA条形码技术,研究茜草药材的"隐性品种"及质量。方法:提取23份茜草饮片的DNA,对ITS、ITS2、psb A-trn H、rbc L和mat K序列扩增和测序,比较扩增测序效率,Dna SP分析变异位点和单倍型,MEGA构建系统发育树,发现茜草药材的"隐性品种",分析"隐性品种"对茜草质量的影响。结果:ITS和ITS2序列由于多拷贝和真菌污染现象,psb A-trn H序列由于高poly结构,扩增和测序成功率均较低。rbc L序列的扩增和测序效率较高,截去引物区,所得序列长度为553bp,共3个变异位点。聚类分析将所有样品分为两个分支,第一分支占73.9%,与正品茜草聚为一支;第二分支形成独立的分支,没有和正品茜草聚为一支,由于这两个分支的茜草药材性状没有明显区别,将未和正品茜草聚成一个分支的样品称为"隐性品种";第一分支的大叶茜草素和羟基茜草素含量明显高于第二分支。结论:DNA条形码可以用于中药"隐性品种"的发现,隐性品种影响药材质量,应加强对中药"隐性品种"的研究。Objective： To study the ‘recessive varieties＇ of Rubiae radix et rhizoma and its influence on quality by DNA barcoding technology. Methods： Genomic DNAs from twenty-three samples of Rubiae radix et rhizoma were extracted. ITS, ITS2, psb A-trn H, rbc L and mat K sequences were amplified and sequenced. PCR amplification efficiency and sequencing efficiency were compared. Variable sites were analyzed by Dna SP, and phylogenetic trees were constructed by MEGA, in order to discover ‘recessive varieties＇ of Rubiae radix et rhizoma and its influence on quality. Results： The PCR amplification and sequencing rate of ITS and ITS2 were low due to polyploid and fungal contamination. The sequencing efficiency of psb A-trn H was low due to poly structure. The length of rbc L was 553 bp with 3 variable sites. Two monophyletic groups were divided by cluster analysis. The first group was gathered with Rubiae radix et rhizoma accounting for 73.9%, while the other one formed a separate group.There was no significant difference of medicinal properties between the two groups. The group which was not clustered with Rubiae radix et rhizoma was ‘recessive varieties＇. The contents of purpurin and rubimaillin in the first group were higher than the other one.Conclusion： DNA barcoding could be used to find ‘recessive varieties＇ of traditional Chinese medicine, which affect the quality of Chinese medicine and should be researched more in future.

关 键 词：中药 隐性品种 条形码 茜草 RBC L 鉴别 

分 类 号：R282.5[医药卫生—中药学]