重齿毛当归ISSR-PCR反应体系的建立与优化  

Establishment and Optimization of ISSR-PCR Reaction System of Angelica pubescence Maxim.

作　　者：郭晓亮 段媛媛 郭杰 游景茂 王澳炎 朱建波 Guo Xiaoliang;Duan Yuanyuan;Guo Jie;You Jingmao;Wang Aoyan;Zhu Jianbo(Sub-center of Chinese Herbal Medicines,Hubei Agricultural Science and Technology Innovation Center,Institute of Chinese Herbal Medicines,Hubei Academy of Agricultural Sciences,Enshi,445000;College of Biological Science and Technologe,Hubei University for Nationalities,Enshi,445000)

机构地区：[1]湖北省农业科学院中药材研究所湖北省农业科技创新中心中药材研究分中心,恩施445000 [2]湖北民族学院生物科学与技术学院,恩施445000

出　　处：《分子植物育种》2022年第8期2682-2688,共7页Molecular Plant Breeding

基　　金：国家现代农业产业技术体系建设专项(CARS-21);湖北省重大专项(2017ACA175);湖北省中央引导地方科技发展专项(2018ZYYD046)共同资助。

摘　　要：本研究在均匀设计的基础上,采用单因素试验进行优化,探寻重齿毛当归进行ISSR-PCR扩增时各组分(即引物,2×Taq Master Mix,模板DNA)的最佳用量,建立重齿毛当归ISSR-PCR扩增体系。筛选的最佳体系为:在20μL的体系中,2×Taq Master Mix:11.4μL、DNA:9.04 ng、引物:0.325μmol/L。同时,还筛选出3条多态性较好、条带稳定的引物(UBC807,UBC810,UBC812)。在此基础上,进行ISSR-PCR扩增验证,结果表明该反应体系扩增效果好,稳定性强,为重齿毛当归的种质资源评价、物种真伪鉴定等提供了一种新的分子标记方法。In order to establish a ISSR-PCR reaction system for Angelica pubescens Maxim,uniform design and single factor design were used to optimize three factors(primer,2×Taq Master Mix,DNA template)in ISSR-PCR reaction system.The results showed that the optimal ISSR-PCR reaction system was as follows:the total 20μL volume included 11.4μL 2×Taq Master Mix,9.04 ng DNA template,and 0.325μmol/L primer.In addition,3 primers(UBC807,UBC810,UBC812)with stable polymorphism and abundant polymorphism were screened.Otherwise,the ISSR amplification verification were performed.The results showed that the optimum reaction system had good amplification effect and strong stability,and provided a new molecular marker method for germplasm resource evaluation and species identification of Angelica pubescens Maxim.

关 键 词：重齿毛当归 ISSR-PCR 体系优化 

分 类 号：S567.239[农业科学—中草药栽培]