Secreted peroxidases VmPODs play critical roles in the conidiation, H2O2 sensitivity and pathogenicity of Valsa mali

Apple Valsa canker, caused by the necrotrophic pathogen Valsa mali, is a devastating disease of apples and causes great financial loss in East Asia. Improving the understanding of apple - V. mali interactions will contribute to disease management. In this study, three predicted secreted peroxidases (VmPOD1, VmPOD2 and VmPOD3) were uncovered based on the secretome and genome information of V. mali. Phylogenetic analysis showed that VmPOD1 is a catalase peroxidase, VmPOD2 is a chloroperoxidase, and VmPOD3 is a plant peroxidase-like peroxidase. The secretion function of the corresponding genes was confirmed using the yeast invertase secretion system. The deletion of VmPODs did not affect the vegetative growth when the mutants (ΔVmPOD1, ΔVmPOD2 and ΔVmPOD3) and the wild-type strain 03-8 were grown on PDA medium at 25 °C in the dark. However, the respective mutants showed impaired conidiation ability with fewer pycnidia, and all gene deletion mutants grew more slowly than 03-8 on PDA supplemented with H2O2 (Final concentration: 0.06 mol/L H2O2). In addition, VmPOD1 and VmPOD2 were found to be significantly up-regulated at an early infection stage, and VmPOD3 showed sustained high expression during the whole infection progress of V. mali. In addition, the virulence of ΔVmPOD3 was significantly reduced, implying that VmPOD3 plays a critical role during the interaction between V. mali and apple. All of the defective phenotypes could be nearly restored by re-introducing the wild-type VmPOD1, VmPOD2 or VmPOD3 allele. The results enhanced our understanding of the secreted peroxidase, which could also act as a type of virulence factor from the necrotrophic pathogen V. mali and provided new insight into the role of the pathogen-secreted peroxidase.