The adenylate cyclase UvAc1 and phosphodiesterase UvPdeH control the intracellular cAMP level, development, and pathogenicity of the rice false smut fungus Ustilaginoidea virens

Abstract

The cyclic adenosine monophosphate (cAMP) signaling pathway plays pleiotropic roles in regulating development and pathogenicity in eukaryotes. cAMP is a second messenger that is important for the activation of downstream pathways. The intracellular cAMP level is modulated mainly by its biosynthesis, which is catalyzed by adenylate cyclases (ACs), and hydrolysis by phosphodiesterases (PDEs). Here, we identified the AC UvAc1 and the cAMP high-affinity PDE UvPdeH in the rice false smut fungus Ustilaginoidea virens; these enzymes are homologs of MoMac1 and MoPdeH in Magnaporthe oryzae (rice blast fungus). A heterogenous complementation assay revealed that UvAc1 and UvPdeH partially or completely rescued the defects in ΔMomac1 and ΔMopdeH mutant M. oryzae. UvAc1 and UvPdeH play important roles in the development and virulence of U. virens. ΔUvac1 and ΔUvpdeH mutant fungi showed defects in conidial production, morphology, and germination; reduced toxicity against germinating rice seeds; and reduced virulence on rice panicles. ΔUvac1 exhibited increased sensitivity to Calcofluor White (CFW) and sodium chloride (NaCl), and decreased sensitivity to Congo Red (CR), while ΔUvpdeH showed increased sensitivity to sodium dodecyl sulfate, CR, sorbitol, and hydrogen peroxide, and decreased sensitivity to CFW and NaCl. High-performance liquid chromatography revealed that the intracellular cAMP level was significantly increased in ΔUvpdeH and decreased in ΔUvac1. Taken together, our results demonstrate that UvAc1 and UvPdeH are conservative components of the cAMP pathway that are important for conidiogenesis, stress responses, virulence, and regulation of the intracellular cAMP level in U. virens.