文献类型：期刊文献

中文题名：黄芪SSR-PCR反应体系的优化及初步应用

英文题名：The optimization and primary application for SSR-PCR Reaction System of the Astragalus

作者：刘亚令[1,2];王文全[2,3];候俊玲[2];张茹[1];杨文玺[4];刘风波[2];魏胜利[2]

第一作者：刘亚令

机构：[1]山西农业大学生命科学学院,山西太谷030801;[2]北京中医药大学中药学院,北京100102;[3]中国医学科学院北京协和医学院药用植物研究所,北京100029;[4]定西师范高等专科学校,甘肃定西743000

第一机构：山西农业大学生命科学学院,山西太谷030801

年份：2014

卷号：25

期号：9

起止页码：2227

中文期刊名：时珍国医国药

外文期刊名：Lishizhen Medicine and Materia Medica Research

收录：北大核心:【北大核心2011】；CSCD:【CSCD_E2013_2014】；

基金：国家工业与信息化部科研项目[No.工信部消费(2011)340];山西农业大学育种基金项目(No.2014YZ10)

语种：中文

中文关键词：黄芪;SSR标记;反应体系优化;多态性

外文关键词：Astragalus; SSR markers; Optimization of reaction system; Polymorphism

摘要：目的为了获得药用植物黄芪SSR-PCR反应的最优体系。方法以山西浑源黄芪为试验材料,采用单因素实验的方法,对影响SSR扩增结果的SSR-PCR反应体系中的5种反应组分(模板DNA、dNTP、TaqDNA聚合酶、引物及退火温度)进行了探索。结果在20μl反应体系中,模板DNA的用量为60 ng,引物的最适浓度为0.6μmol/L,dNTP浓度为0.1 mmol/L、TaqDNA聚合酶浓度为0.05 U/μl、退火温度为57℃为最适反应体系。利用此反应体系对不同来源(山西、黑龙江、辽宁、河北、甘肃)的黄芪进行PCR扩增及琼脂糖凝胶电泳检测,扩增结果清晰且谱带多态性丰富。结论成功建立了药用植物黄芪的SSR-PCR最优体系,并利用筛选出的引物检测出不同产地黄芪具有很高的多态性,为今后黄芪种质资源的分子评价奠定了技术基础。
Objective To get the optimal and application SSR-PCR reaction system of the Astragalus. Methods The main ingredients of the reaction system were studied by the method of single factor experiment,five factors including template DNA,dNTP,TaqDNA and primers as well as annealing temperature were screened and optimized. Results the best combination of the20μL reaction system was the optimal concentration of template DNA for 60 ng/μL,primers for 0. 6 μmol/L,the optimal concentration of dNTP for 1. 0mmol/L,the perfect dosage of TaqDNA polymerase enzyme for 0. 5U/μL when the annealing temperature was 57℃. Which using of this reaction system to do PCR amplification with different sources of Astragalus（ Shanxi province,Honglongjaing province,Liaoning province,Hebei province,Gansu province） and then detected it by PAGE electrophoresis and made sure that the optimal SSR-PCR reaction system was stable and repeatable. Conclusion Succeeding establish the optimal system of SSR-PCR medicinal plants of Astragalus,and obtain the highly polymorphism using the selected primers to detect different origin of Radix Astragalus,which showed the optimization SSR system and provided a technical support for future molecular evaluation of germplasm resources of Astragalus.