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利用分别与抗稻瘟病基因Pi-1和Pi-9紧密连锁的SSR标记MRG4766和SNP标记FP1+FP2+RP组成双重PCR体系,通过设置Mg2+、d NTPs和引物浓度梯度,探索双重PCR体系中各反应物的使用量,由此确定最优双重PCR反应体系.在水稻恢复系R1059与R673杂交的BC3F2群体中随机挑选20个单株对该双重PCR反应体系的效果进行验证.结果表明,20个单株的双重PCR扩增结果与2个单一PCR扩增结果相符,并且条带清晰,说明该双重PCR反应体系在利用分子标记进行基因聚合上可提高选择效率和减少费用.
The double PCR system was constructed by using SSR marker MRG4766 and SNP marker FP1 + FP2 + RP which were closely linked to the genes Pi-1 and Pi-9, respectively. Through setting Mg2 +, dNTPs and primer concentration gradient, And the amount of reactants used to determine the optimal duplex PCR reaction system.The results of the double PCR reaction system were validated by randomly selecting 20 plants in the BC3F2 population crossed with the R1059 and R673 rice restorer lines.The results showed that 20 The results of double PCR amplification of single plant were in good agreement with the results of two single PCR amplification and the bands were clear. This double PCR reaction system can improve the selection efficiency and reduce the cost when using the molecular marker for gene polymerization.