树莓SSR体系的建立及优化  

Establishment and Optimization of SSR System in Raspberry

作　　者：李媛媛[1] 郭修武[1] 代汉萍[1] 刘海涛[1] 

机构地区：[1]沈阳农业大学园艺学院,沈阳110161

出　　处：《沈阳农业大学学报》2009年第4期478-480,共3页Journal of Shenyang Agricultural University

基　　金：农业部科技部行业科研专项(nykyzx07-028);"十一五"农业部948项目(G-25)

摘　　要：为建立适宜树莓(Rubus idaeus L.)的SSR分析体系,设计Taq DNA聚合酶、Primer、Mg2+、dNTP和DNA的5因素4水平正交试验,对SSR反应体系进行筛选和优化。结果表明:适宜树莓的SSR分析体系为15μL反应体系中含Taq DNA聚合酶0.5U,Primer 0.2μmol·L-1,Mg2+0.9mmol·L-1,dNTP 0.25 mmol·L-1,DNA 60ng;Primer Rubus285a的最佳退火温度为54～59℃。利用此反应体系对部分树莓品种进行SSR-PCR扩增并电泳检测,扩增谱带清晰且多态性较好,证明此体系稳定可靠。Orthogonal experiment with five factors including Taq polymerase,primer,Mg2＋,dNTP and as template DNA at four content levels was designed and optimized in order to establish SSR reaction system in raspberry.The results showed that 0.5 U Taq polymerase,0.2 μmol·L-1 Primer,0.9 mmol·L-1 Mg2＋,0.25 mmol·L-1 dNTP,60 ng template DNA were the optimal conditions in the reaction of 15 μL volume.The optimal annealing temperature of primer Rubus 285a ranged from 54℃ to 59 ℃.Under optimal conditions,SSR reactions were carried out with 10 cultivars of raspberries and tested with electrophoresis.The results revealed clear and high polymorphism which demonstrated that the reaction system established in this experiment was stable and reliable.

关 键 词：树莓 正交试验 SSR 优化 

分 类 号：S663.2[农业科学—果树学]