摘要： 为建立食用向日葵分子标记反应体系,以食用向日葵四叶期叶片为DNA模板提取材料,采用单因素试验和正交试验设计,对SSR-PCR反应体系中的6因素(10×PCR Buffer、Mg²⁺ 、dNTPs、引物、 Taq DNA 聚合酶和DNA模板)在5水平上进行正交优化试验,并比较了不同浓度Mg²⁺、 Taq DNA聚合酶、模板DNA对扩增效果的影响,结果表明,各因素水平变化对反应体系的影响为Mg²⁺ > Taq DNA 聚合酶(引物)>DNA模板>10×PCR Buffer>dNTPs。最终建立食用向日葵SSR-PCR 最佳反应体系为:在总体系为20 μL的SSR-PCR反应体系中包括10×PCR Buffer 0.2 mmol/L、Mg²⁺2.0 mmol/L、dNTPs 1.8 mmol/L、Taq DNA聚合酶 0.2 U、DNA 50 ng、引物1.5 mmol/L。

关键词: 食用向日葵, SSR-PCR, 体系优化

Abstract: In order to established confectionary sunflower molecular markers reaction system, the study assuming DNA template from four-leaf stage of confectionary sunflowers leaves for experimental materials, this study used the single factor test and orthogonal design for optimizing six SSR-PCR reaction system factors (10×PCR Buffer, Mg²⁺, dNTPs, primers, Taq DNA polymerase, DNA template and compared different concentrations of Mg²⁺, dNTPs, Taq DNA polymerase and DNA template amplification effect of the impact in SSR-PCR reaction system.The result showed that the range of the influence of six factors in the reaction system was Mg²⁺> Taq DNA polymerase (primer)>DNA template>10×PCR Buffer>dNTPs.And this study established confectionary sunflower SSR-PCR reaction system(20 μL):10×PCR Buffer 0.2 mmol/L, Mg²⁺ 2.0 mmol/L, dNTPs 1.8 mmol/L, Taq DNA polymerase 0.2 U, DNA 50 ng, primer 1.5 mmol/L.

Key words: Confectionary sunflower, SSR-PCR, Reaction system

中图分类号: 

• S565.5