巴西橡胶树棒孢霉落叶病菌Hog1同源基因的克隆和序列分析

Cloning and Sequence Analysis of Hog1 Gene from Corynespora cassiicola of Hevea brasiliensis

为了解MAPK信号途径中hog1基因在多主棒孢病菌(Corynespora cassiicola)侵染橡胶树过程中的作用,根据几种丝状真菌的hog1基因设计简并引物,采用PCR和RT-PCR的方法扩增巴西橡胶树棒孢霉落叶病菌CC01的hog1基因,并对其扩增产物进行测序,同时还对该基因编码的蛋白进行氨基酸序列比对和系统发育分析。结果表明:该基因开放阅读框架为915 bp,编码304个氨基酸残基,包含6个内含子;该基因的氨基酸序列与小麦黄斑叶枯病菌(Pyrenophora tritici)MAPK HOG1(XP_001935555.1)、谷子弯孢病菌(Cochliobolus lunatus)MAP kinase ClK1(AFJ42499.1)等高度同源。

In order to understand the roles of hogl Gene in invasion process of Corynespora cassiicola into Hevea brasiliensis, hogl gene of CC01 was amplified by PCR and RT-PCR methods and the degenerate primers were designed according to hogl gene of several filamentous fungi. The amplification products were sequenced, the amino acid sequence were aligned and phylogenetic clustering analysis were done. Sequence analysis extrapolated that the open reading frame of this gene was 915 bp with 6 introns, encoding a putative protein of 304 amino acids. Phylogenetic clustering suggested that the sequence of CCHOG1 showed higher similarity to MAPK HOG1 of Pyrenophora tritici , MAP kinase C1K1 of Cochliobolus lunatus.