蓝莓SCoT标记分析体系的建立与优化

Establishment and Optimization of SCoT System in Blueberry

为建立蓝莓SCoT标记分析体系,为蓝莓的分子育种和遗传多样性分析等研究提供技术支持,以蓝莓叶片为试材,采用L16(45)正交设计方法,对影响蓝莓SCoT-PCR反应的Mg2+、dNTPs、Taq DNA聚合酶、模板DNA及引物等因素进行优化筛选,建立了适用于蓝莓遗传分析的SCoT-PCR扩增体系,即20μL的反应体系中含有:Mg2+为2.813 mmol/L、dNTPs为0.100 mmol/L、Taq酶为1.5 U、Primer为0.2μmol/L、DNA为10 ng。应用优化的反应体系,对32个SCoT引物和6个蓝莓品种(‘Centurion’、‘Premier’、‘Homebell’、‘O'Neal’、‘Tifblue’、‘Misty’)进行扩增,获得了条带清晰、稳定可靠的电泳结果。

The study aims to establish an optimal reaction system of SCOT-PCR in blueberry and provide technical support for molecular breeding and genetic diversity analysis. Blueberry was used as material, orthogonal design method was applied to optimize SCoT-PCR amplification system of blueberry in five factors such as Mg2＋, dNTPs, Taq DNA polymerases, template DNA and primers at five levels. An optimal SCoT-PCR amplification system （20 μL） of blueberry was established, i.e. Mg2＋ 2.813 mmol/L, dNTPs 0.100 mmol/L, Taq polymerase 1.5 U, primer 0.2 μmol/L, template DNA 10 ng. 32 SCoT primers and six blueberry varieties （＇Centurion＇, ＇Premier＇, ＇Homebell＇, O＇Neal＇, ＇Tifblue＇, ＇Misty＇）were analyzed with the optimized reaction system. The study indicated that amplified results with fine diversity and clear strap were gained.