摘要： 采用L16(4)正交设计和二因素完全随机试验,分析了月季SSR-PCR反应体系的主要成分Taq DNA聚合酶、样本浓度、Mg²⁺浓度、dNTP浓度以及引物浓度对扩增结果的影响,建立了适合月季的SSR反应体系.研究结果表明:在20μL反应体系中,Taq DNA聚合酶宜加入1.U,样本最适宜浓度为30～40 ng,Mg²⁺的最适浓度为1～1. mmol/L,dNTP最适浓度为0.2 mmol/L,单引物的最适浓度均为1μmol/L.用建成的反应体系对19对引物筛选,对12个月季品种扩增,3%的琼脂糖凝胶电泳检测,品种间DNA谱带多态性丰富,证明该体系稳定可靠.

关键词: 月季, SSR, 正交试验, 反应体系

Abstract: Using orthogonal design, five factors including concentration of template、 Mg²⁺、 dNTP, primers and Taq DNA polymerase were optimized to establish SSR analysis system in roses.The results showed that 30- 40 ng DNA tem - plate, 1- 1. 5 mmol/L Mg²⁺, 0. 2mmol/ L dNTP, 1 L mol/ L each primer and 1. 5U Taq DNA polymerase in 20 L L reac - tion system were the optimal option.Through the optimized reaction system, SSR fragments of 12 rose cultivars were ob - tained and detected. Polymorphism between different cultivars was abundantly detected by 3% agarose gels,which showed that the system established in this study was suitable and stable.

Key words: Rose, SSR, Orthogonal design, System optimization

中图分类号: 

• S685.12