无患子SRAP-PCR反应体系的优化

建立适合无患子Sapindus mukurossi的相关序列扩增多态性-聚合酶链式反应(SRAP-PCR)最佳反应体系,为研究无患子种质资源的遗传多样性、亲缘关系及遗传改良等奠定基础。以南平产的无患子叶片为材料,采用单因素法对体系中的DNA模板、镁离子、三磷酸碱基脱氧核苷酸(dNTPs),引物和TaqDNA聚合酶等5因素进行优化。确立了适合无患子SRAP的最佳反应体系(20μL):1×PCR缓冲液,50 ng模板DNA,2.0 mmol·L-1镁离子,0.2 mmol·L-1dNTPs,0.5×16.67 nkat(0.5 U)TaqDNA聚合酶,0.4μmol·L-1引物。利用该体系对12份无患子种源材料进行PCR扩增,获得了清晰、稳定的DNA谱带,说明优化后的体系适宜无患子地理种源的遗传多样性分析。

Optimization of SRAP-PCR in Sapindus mukurossi

Establish an optimal sequence-related amplified polymorphism（SRAP）-PCR reaction system was to lay the foundation of studying genetic diversity, relationship analysis and genetic improvement in Sapindus mukurossi. The leaves from Nanping, Fujian Province were as material. Five influence factors including template DNA, Mg²⁺, dNTPs, primers and TaqDNA polymerase were optimized with single factor method．The optimum reaction system by the volume of 20 μL was established as follows: 1×PCR buffer, 50 ng DNA template, 2.0 mmol·L^－1 Mg²⁺, 0.2 mmol·L^－1 dNTPs, 0.5×16.67 nkat TaqDNA polymerase and 0.4 μmol·L^－1 primers. Using the optimization system, 12 different source materials of Sapindus mukurossi were detected a clear and stable results. It shows that the optimazed SRAP-PCR system could be suitable for the genetic diversity analysis of Sapindus mukurossi geographical provenance. ［Ch, 7 fig. 3 tab. 19 ref.］