番茄抗性基因Ty-2和Ph-3多重PCR的检测应用

Application of Multiple PCR for Ty-2 and Ph-3 Genes in Tomato

创建可以同时检测番茄抗黄化曲叶病毒病Ty-2基因和抗晚疫病Ph-3基因分子标记的多重PCR体系,以快速准确地鉴定筛选到目标基因型。采用多重PCR结合CAPS技术,同时加入2对引物T0302和TG328,进行扩增及酶切反应,2%琼脂糖凝胶电泳检测,获得不同基因型相应的带型,从而建立多重PCR及产物酶切体系,优化此体系后再进行验证与应用。该多重PCR体系可靠高效,显示共有9种带型对应Ty-2和Ph-3基因相应的基因型,与采用单引物普通PCR技术分别鉴定的基因型结果一致,可以用于番茄分子标记辅助育种。

We established a multiplex PCR system to identify Ty-2 and Ph-3 resistance genes of tomato simultaneously,for rapid and accurate identification and screening of target genotype.Multiplex PCR-CAPS technology was used to amplify and react with two pairs of primers T0302 and TG328.2%agarosegel electrophoresis was used to detect the corresponding bands to differentiate the genotype of the identified materials,and to establish multiplex PCR system.The results showed that there were 9 kinds of bands corresponding to the genotypes,which were consistent with the genotype identified by single primer PCR.The multiplex-PCR systems are reliable and efficient,it could be very useful for marker-assisted selection during early stage in tomato and speed up breeding procedure.