四引物扩增受阻突变体系PCR在苦瓜抗白粉病分子标记开发中的应用

Application of Tetra-primer ARMS PCR in Development of Molecular Markers for Powdery Mildew Resistance of Bitter Melon

以苦瓜高抗、高感白粉病的父母本及其F2代为试材,根据双本测序比对获得的SNP突变位点来设计四引物ARMS PCR分子标记引物。采用正交设计L16(44)方法对模板DNA、dNTPs、Mg2+、Taq DNA聚合酶4个因素和内外引物的浓度比进行体系优化,并利用优化后的体系对引物进行筛选。研究了适用于苦瓜的四引物ARMS PCR分子标记开发及引物筛选体系,以期为苦瓜抗白粉病分子标记辅助选择提供参考依据。结果表明:对优化后的四引物ARMS PCR体系验证,能正确地在父母本及其F2代中分型,优化后的体系适用于苦瓜抗白粉病的四引物ARMS PCR分子标记的筛选。

Parental lines with high bitter gourd resistance and susceptible powdery mildew and its F2 generation were used as test materials.The tetra-primer ARMS PCR molecular marker primers were designed according to the SNP mutation sites obtained by double-sequence sequencing.Orthogonal design L16（44）method was used to optimize the system of template DNA,dNTPs,Mg2 ＋,Taq DNA polymerase and internal and outer-primer concentration ratio,and then the optimized system was used to screen the primers.The tetra-primer ARMS PCR molecular marker development were suitable for bitter melon and primer screening system were studied in order to provide reference for molecular marker-assisted selection of bitter melon-resistant powdery mildew.The results showed that the optimized the tetra-prime ARMS PCR system could be correctly typed in parents and their F2 generation and was suitable for the screening of the tetra-prime ARMS PCR molecular marker which was against powdery mildew.