巴西橡胶树SSR反应体系的优化

摘 要： 以巴西橡胶树无性系针选2号为试验材料,对橡胶树SSR-PCR反应的5个主要因素：TaqDNA聚合酶、Mg2+、引物、DNA模板及dNTPs进行优化试验。筛选出了各反应因素的最佳水平，建立了橡胶树SSR-PCR最佳反应体系（20ul）: TaqDNA聚合酶0.8U；Mg2+ 浓度1.5mmol•L-1；引物浓度0.5μmol•L-1；模板DNA 含量50ng； dNTPs浓度 0.25 mmol•L-1。并利用此反应体系,用引物1对14个巴西橡胶树品种进行SSR分析,8%的非变性聚丙稀酰胺凝胶电泳检测显示不同品种间DNA谱带多态性丰富，结果证明，此反应体系是稳定可靠的，这为下一步进行的遗传多样性分析、指纹图谱构建等研究奠定基础

Optimization on SSR Reaction System of Hevea brasiliensis

Abstract: Five main factors which affected SSR-PCR reaction: TaqDNA polymerase, Mg2+, primer, template DNA and dNTPs were studied with the Hevea brasiliensis plant material Zhenxuan 2 to screen the optimal reaction system. It was indicated by the results that the optimal reaction system of SSR-PCR for Hevea brasiliensis was 20 ul of total reaction volume containing 0.8 units of TaqDNA polymerase, 1.5mmol•L-1 of Mg2+, 0.5μmol•L-1 of primer, 0.25 mmol•L-1 of dNTPs and 50 ng template DNA. Fourteen plant materials, one primer pair and 8% polyacrylamidegel were selected to detect the reaction system. The results seemed that the optimal system was stable and credible which can build the basis on further study, e.g. genetic diversity assessment and construction of fingerprint etc. Key words: Hevea brasiliensis, simple sequence repeat (SSR),optimization