大丽轮枝菌 VdCPMO基因克隆与功能分析

大丽轮枝菌 Verticillium dahliae是一种典型的土传病原真菌,可侵染400多种植物引致黄萎病,造成巨大的经济损失。微菌核是大丽轮枝菌的特殊休眠结构,能够在土壤中存活14年之久,是病害的主要初侵染来源。本研究在前期微菌核萌发表达谱的基础上,选择上调倍数最高的环戊酮1,2-单加氧酶基因（ VDAG_03943）,进行克隆与功能分析。结果表明,该基因全长1 929bp,cDNA序列全长1 668bp,编码555个氨基酸组成的蛋白,命名为 VdCPMO。与野生型菌株JY相比,敲除突变体菌株的菌落生长减慢,微菌核萌发率显著降低,芽管平均长度明显较短,而互补突变体菌株与野生型菌株JY间无显著性差异。致病力测定结果显示,敲除突变体菌株的微菌核丧失了对棉花的致病力。

Cloning and functional analysis of VdCPMO gene in Verticillium dahliae

Verticillium dahliae, a typical soilborne fungus, infects over 400 plant species and causes huge economic loss. Microsclerotia can survive for 14 years in the soil as dormancy structures and serve as the primary inoculum on many hosts. According to the whole genome-wide expression profiles on the germination of microsclerotia, the VDAG_03943 gene, coding cyclopentanone 1,2-monooxygenase, was most highly expressed by the fungus during the germination process. In the present study, we cloned the sequence and characterized the function of VDAG_03943. Sequence analysis showed that VDAG_03943, named VdCPMO, has a full length of 1 929bp. The cDNA coding region of VdCPMO contains 1 668bp of nucleotides that encode a protein consisted of 555 amino acids. The results displayed that deletion mutant strains exhibited several developmental defects including slower growth of colony, reduced germination rate of microsclerotia, and reduced average length of germ tube as compared with the wild type strain. There were no significant differences between the complementary mutant strain and wild type JY strain. Furthermore, the microsclerotia of deletion mutant strains lost the pathogenicity to cotton.