十字花科蔬菜黑斑病菌的PCR鉴定

PCR identification of the pathogens causing black leaf spot of cruciferae vegetables

在对十字花科蔬菜黑斑病菌(Alternariasp.)3个种及相近种的5·8SrDNA和其侧翼ITS区进行测序的基础上,分别设计合成了鉴定白菜黑斑病菌3个种的特异性引物。PCR扩增结果表明:Abre1和Abre2引物对能特异性扩增芸苔链格孢(A·brassicae)371bp的片段,Abra1和Abra2引物对能特异性扩增甘蓝链格孢(A·brassicicola)457bp的片段,Ajap1和Ajap2引物对能特异性扩增萝卜链格孢(A·japonica)411bp的片段,而且其它近源种未扩增出目标片段,说明这3个引物对可以作为十字花科蔬菜黑斑病菌3个种快速检测鉴定的分子特征标记。

Based on sequencing of the regions coding for 5.8S rDNA and the flanking internal transcribed spacers(ITS1 and ITS2) from 37 domestic and foreign isolates belonging to three species of Alternaria brassicae, A.brassicicola, A.japonica and the related species, three specific primer pairs are designed. The PCR assay shows that the primer pairs Abre1 and Abre2 can amplify specifically the 371 bp fragment of A. brassicae, Abra1 and Abra2 primer pairs for 457 bp fragment of A.brassicicola specifically and Ajap1 and Ajap2 primer pairs for 411 bp fragment of A.japonica. These results show that the three primer pairs can be used as a molecular marker and to separately identified the three species by PCR method.