Apple canker disease, caused by Valsa mali, is one of the most serious apple tree diseases in China. VmSom1 is an important transcription factor that acts on the cyclic adenosine signaling pathway (cAMP/PKA), regulating the growth, development, morphological differentiation, and pathogenic forces of the pathogen. We perform transcriptome analysis of the VmSom1 deletion mutant and the wild-type strain 11-175 and identify a significantly differentially expressed gene, VM1G_06867, a zinc finger motif transcription factor in V. mali. In this study, we obtain the VM1G_06867 gene using the single deletion mutant via homologous recombination. To determine the relationship between VmSom1 and VM1G_06867, we also obtain a double deletion mutant ΔVmSom1/06867. Compared to the wild-type strain 11-175, the single deletion mutant VM1G_06867 shows a drastic reduction in growth rate and forms more pycnidia on the PDA medium. Additionally, the growth of the mutant is inhibited by SDS, Congo red, and fluorescent brighteners. In comparison to the single deletion mutant VmSom1, the double deletion mutant ΔVmSom1/06867 shows no significant change in growth or conidiation and is unable to produce conidia. The growth rate is significantly increased in Congo red, NaCl, and Sorbitol mediums. These results demonstrate that VM1G_06867 plays important roles in growth, pathogenicity, asexual development, and maintenance of cell wall integrity. VM1G_06867 can recover osmotic stress and cell wall integrity defects caused by the deletion of VmSom1, as well as restore the loss of pathogenicity caused by the deletion of the VmSom1 gene, but not completely.

Carbon catabolite repression (CCR) is a very important mechanism for efficient use of carbon sources in the environment and is necessary for the regulation of fungal growth, development, and pathogenesis. Although there have been extensive studies conducted regarding this mechanism in fungi, little is yet known about the effects of CreA genes on Valsa mali. However, based on the results obtained in this study for the identification of the VmCreA gene in V. mali, it was determined that the gene was expressed at all stages of fungal growth, with self-repression observed at the transcriptional level. Furthermore, the functional analysis results of the gene deletion mutants (ΔVmCreA) and complements (CTΔVmCreA) showed that the VmCreA gene played an important role in the growth, development, pathogenicity, and carbon source utilization of V. mali.

Apple Valsa canker disease, caused by Valsa mali, is one of the most destructive diseases of apple trees in China and seriously threatens the sustainable development of the apple industry. VmSom1 acts as a core transcription factor in the cyclic adenosine monophosphate/protein kinase A (cAMP/PKA) signaling pathway and regulates vegetative growth, development and pathogenicity of this phytopathogen. Transcriptome analysis was performed using the VmSom1 deletion mutant and the wild-type strain sdau11-175, and a significantly differentially expressed transcription factor, VmGAL4, was identified. In this study, the single-gene deletion mutant ΔVmGAL4 and the double-gene deletion mutant ΔVmSom1/VmGAL4 were constructed via homologous recombination, aiming to preliminarily explore the interaction between these two genes. Sequence analysis revealed that the VmGAL4 protein contains a conserved fungal_TF_MHR domain spanning amino acids 164 to 614. Phylogenetic analysis indicated that VmGAL4 shares the closest phylogenetic relationship with homologs from Cytospora schulzeri and Cytospora chrysosperma. Phenotypic assays demonstrated that the VmGAL4 deletion mutant exhibited markedly reduced mycelial growth rate and fewer pycnidia production. Additionally, the mutant displayed enhanced sensitivity to cell wall inhibitors and osmotic stress agents, along with significantly increased capacity to utilize various carbon and nitrogen sources and decreased pathogenicity compared with the wild-type strain. Compared with the single deletion mutant ΔVmSom1, the double mutant ΔVmSom1/VmGAL4 partially rescued the growth defects and also alleviated the reduction in pathogenicity to a certain extent. Nevertheless, conidial production remained severely inhibited in the double mutant. Collectively, VmGAL4 is involved in the regulation of vegetative growth, asexual development, cell wall integrity, osmotic stress response, carbon and nitrogen source utilization, and pathogenicity in V. mali.