OPY2 is a highly conserved membrane signal-sensing protein in fungi, yet its function in Fusarium verticillioides, the causal agent of maize ear rot, remains unclear. In this study, we generated an FvOPY2 knockout mutant and found that FvOPY2 localizes to the plasma membrane and intracellular vesicles. FvOPY2 deletion caused reduced growth, sporulation defects, hypersensitivity to cell wall stressors, and significantly decreased fumonisin production and maize ear pathogenicity. Quantitative  reverse transcription polymerase chain reaction (qRT-PCR) revealed that FvOPY2 regulated the expression of the chitin synthase gene family and the fumonisin biosynthetic gene cluster. These findings demonstrate that FvOPY2 regulates chitin and fumonisin biosynthesis and maize ear infection, potentially acting as a signal receptor for environmental sensing. This study provides a theoretical basis and novel strategies for developing FvOPY2 as a target for new fungicides.