当归简单重复序列区间-聚合酶链反应体系的建立及优化与品种(系)间遗传关系研究  

Establishment and Optimization of ISSR-PCR Reaction System for Angelica sinensis and Genetic Relationship Study of Different Varieties or Strains

作　　者：朱田田[1,2] 张裴斯[1] 晋玲[1] 刘效瑞[3] 李应东[4] 刘进[1] 

机构地区：[1]甘肃中医学院中(藏)药资源研究所,兰州730000 [2]甘肃中医学院药用植物遗传育种研究所,兰州730000 [3]甘肃定西市旱作农业科研推广中心,甘肃定西743000 [4]甘肃中医学院附属医院,兰州730000

出　　处：《中国药房》2014年第35期3265-3269,共5页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81360605);"十二五"国家科技支撑计划课题(No.2011BAI05B02);甘肃省自然科学基金资助项目(No.1308RJZA127)

摘　　要：目的:建立并优化当归简单重复序列区间(ISSR)-聚合酶链反应(PCR)体系并对6个当归品种(系)的遗传关系进行研究。方法:以ISSR-PCR扩增结果为指标,Mg2+浓度、TaqDNA聚合酶活性、dNTPs浓度、引物浓度、DNA模板用量为5因素,通过正交、单因素试验优化ISSR-PCR体系。应用优化体系对6个不同品种(系)的当归样本进行遗传关系分析。结果:当归ISSR-PCR最佳反应体系为在20μl反应体系中含Mg2+(10×PCR Buffer)1.5 mmol/L、TaqDNA聚合酶0.6 U、dNTPs 0.375 mmol/L、引物0.3μmol/L、DNA模板42 ng。6个当归品种(系)的多态性位点百分率为13.04%,Nei’s基因多样性指数为0.058 0,Shannon多样性指数为0.082 7。结论:建立的当归ISSR-PCR体系可用于当归分子遗传学研究,6个当归品种(系)间遗传差异较小。OBJECTIVE：To establish and optimize ISSR-PCR reaction system for Angelica sinensis. METHODS：The ISSRRCR reaction system were optimized by orthogonal design and single-factor design with ISSR-PCR amplification as index using Mg2＋concentration,TaqDNA polymerase activity,dNTPs concentration,primer concentration,template DNA concentration as factors. The genetic relationship of 6 varieties or strains of A. sinensis were analyzed by optimized system. RESULTS：The optimum reaction system（20 μl）contained 1.5 mmol/L Mg2＋（10×PCR Buffer）,0.6 U TaqDNA polymerase,0.375 mmol/L dNTPs,0.3 μmol/L ISSR primer and 42 ng/20 μl DNA template. The average percentage of polymorphie bands（PPB）of 6 different varieties or strains of A. sinensis was 13.04%,Nei＇s genetic diversity index（He）and Shannon＇s information index（Ⅰ）were 0.058 0 and 0.082 7.CONCLUSIONS：The established ISSR-PCR reaction system can be used to study molecular genetics of A. sinensis. The genetic differences among 6 varieties or strains of A. sinensis are small.

关 键 词：当归 简单重复序列区间聚合酶链反应体系 建立 遗传关系 

分 类 号：R282[医药卫生—中药学] R932[医药卫生—中医学]