黄芪ISSR-PCR反应体系的建立及优化  

Establishment and Optimization of ISSR-PCR Reaction System for Astragalus

作　　者：张茹[1] 刘亚令[1] 梁建萍[1] 张鹏飞[2] 

机构地区：[1]山西农业大学生命科学学院,山西太谷030801 [2]山西农业大学园艺学院,山西太谷030801

出　　处：《山西农业科学》2014年第1期9-11,共3页Journal of Shanxi Agricultural Sciences

基　　金：山西省科技攻关项目(20110313002-2;20130311022-6);山西农业大学科技创新基金项目(2010003)

摘　　要：为获得最优黄芪ISSR-PCR反应体系,以山西省浑源县官儿乡蒙古黄芪为试验材料,利用单因素试验法对反应体系中的4个因素(模板DNA,dNTPs,Taq DNA聚合酶,ISSR引物)分别设置浓度梯度进行体系优化。结果表明,20μL黄芪ISSR-PCR最优体系为:模板DNA质量浓度6.0 ng/μL,dNTPs浓度0.03 mmol/L,Taq DNA聚合酶浓度0.05 U/μL,引物浓度0.80 mmol/L。最后用不同产地的黄芪检测该体系,结果发现,PCR产物多态性良好。优化体系的确立为今后黄芪种质资源的分子评价奠定了技术基础。To get the optimization ISSR-PCR reaction system of Astragalus,the ISSR-PCR system through the single factor experiment was established and optimized.The results show that the best reaction system is a 20 μL volume ISSR-PCR system with DNA temphte of 6.0 ng/μ L,dNTPs of 0.03 mmol/L,TaqDNA polymerase of 0.05 U/μL,primer of 0.80 mmol/L and the annealing temperature of 49 ℃.Finally,concluded that this optimal system of ISSR-PCR was established successfully after tested using the Astragalus membranaceus from provinces of Hebei,Jilin,Heilongjiang,Shanxi,Inner-mongoulia,Shandong,Shaanxi,and Gansu.

关 键 词：黄芪 梯度 ISSR PCR 

分 类 号：S567.239[农业科学—中草药栽培]