Verticillium dahliae Kleb is a typical soilborne pathogen that can cause vascular wilt disease on more than 400 plants. Functional analysis of genes related to the growth and virulence is crucial to revealing the molecular mechanism of the pathogenicity of V. dahliae. Glycosidase hydrolases can hydrolyze the glycosidic bond, and some can cause host plant immune response to V. dahliae. Here, we reported a functional validation of VdGAL4 as an α-galactosidase that belongs to glycoside hydrolase family 27. VdGAL4 could cause plant cell death, and its signal peptide plays an important role in cellular immune response. VdGAL4-triggered cell death depends on BAK1 and SOBIR1 in Nicotiana benthamiana. In V. dahliae, the function of VdGAL4 in mycelial growth, conidia, microsclerotium, and pathogenicity was studied by constructing VdGAL4 deletion and complementation mutants. Results showed that the deletion of VdGAL4 reduced the conidial yield and conidial germination rate of V. dahliae and changed the microscopic morphology of conidia; the mycelia were arranged more disorderly and were unable to produce microsclerotium. The VdGAL4 deletion mutants exhibited reduced utilization of different carbon sources, such as raffinose and sucrose. The VdGAL4 deletion mutants were also more sensitive to abiotic stress agents of SDS, sorbitol, low-temperature stress of 16°C, and high-temperature stress of 45°C. In addition, the VdGAL4 deletion mutants lost the ability to penetrate cellophane and its mycelium were disorderly arranged. Remarkably, VdGAL4 deletion mutants exhibited reduced pathogenicity of V. dahliae. These results showed that VdGAL4 played a critical role in the pathogenicity of V. dahliae by regulating mycelial growth, conidial morphology, and the formation of microsclerotium. IMPORTANCE This study showed that α-galactosidase VdGAL4 of V. dahliae could activate plant immune response and plays an important role in conidial morphology and yield, formation of microsclerotia, and mycelial penetration. VdGAL4 deletion mutants significantly reduced the pathogenicity of V. dahliae. These findings deepened the understanding of pathogenic virulence factors and how the mechanism of pathogenic fungi infected the host, which may help to seek new strategies for effective control of plant diseases caused by pathogenic fungi.
Keywords: Verticillium dahliae; conidia; glycosidase hydrolases; glycoside hydrolases; microsclerotia; pathogenicity.