AbstractRice false smut caused byUstilaginoidea virensis one of the widespread rice diseases across the globe in recent years, however, we know little about its molecular mechanism of infection. The cAMP signaling pathway functions directly in the development and formation of infectious structures to regulate the infection process in many pathogenic fungi. In order to investigate the role of the cAMP signaling pathway inU. virens, UvCap1, a cyclase-associated-protein homologous toSaccharomyces cerevisiaeSrv2 was identified. Three targeted deletion mutants of theUvCAP1gene were obtained with gene replacement strategy assisted with CRISPR-Cas9 system. TheUvCAP1deletion mutants showed defects in mycelial growth and conidial production. Inoculation experiments demonstrated that ΔUvcap1exhibited defects in pathogenicity. Compared with the wild-type strain, ΔUvcap1showed decreased tolerance to sorbitol and H2O2, and increased tolerance to NaCl, CFW and SDS, and the intracellular cAMP level was significantly reduced in ΔUvcap1. Yeast two-hybrid assay identified the interactions of UvCap1 with UvAc1 (adenylase cyclase), two Ras proteins (UvRas1 and UvRas2) and UvSte50. Taken together, as a component of cAMP signaling pathway, UvCap1 plays important roles in the development and pathogenicity ofU. virens.