Roles of NADPH oxidases in regulating redox homeostasis and pathogenesis of the poplar canker fungus Cytospora chrysosperma

Poplar canker, caused by the fungus Cytospora chrysosperma, results in tremendous losses in poplar plantations in China. Although NADPH oxidases (NOXs) play important roles in the development and pathogenicity of several pathogenic fungi, their roles in C. chrysosperma remain unclear. In this study, we characterized three NOX genes (CcNox1, CcNox2, and CcNoxR) in C. chrysosperma. All three genes were highly upregulated during poplar branch infection, and deletion of any of them severely reduced virulence on poplar branches. Furthermore, deletion of either CcNox1 or CcNoxR resulted in a significant increase in endogenous reactive oxygen species production in hyphae, enhanced influx of Ca²⁺, the disruption of redox homeostasis and compromised mitochondrial integrity. Moreover, biosynthesis and secretion of a known virulence factor oxalic acid was obviously defective and exogenous oxalic acid supplementation rescued the virulence of the mutants. Taken together, our findings reveal that NOXs play important roles in redox homeostasis, mitochondrial integrity and pathogenicity in C. chrysosperma.