Fusarium crown rot, a widespread and destructive disease affecting cereal crops (particularly wheat and barley), is primarily caused by the soil-borne fungal pathogen Fusarium pseudograminearum. Secondary metabolites (SMs) play a crucial role in colonization and host tissue invasion by pathogenic fungi. In this study, we investigated the functional role of FpTox2, a secondary metabolite-related gene in F. pseudograminearum. An FpTox2 deletion mutant exhibited significantly reduced radial growth compared to wild-type F. pseudograminearum. Notably, the mutant strain completely lost conidiation capacity under induced conditions. Furthermore, although it showed decreased sensitivity to the cell membrane inhibitor sodium dodecyl sulfate (SDS), the mutant demonstrated enhanced susceptibility to NaCl, a metal ion stressor. Most importantly, the pathogen’s virulence was markedly attenuated in wheat stem base infections following FpTox2 deletion, and we demonstrated that FpTox2 regulates pathogen virulence by influencing deoxynivalenol production. In conclusion, FpTox2 is crucial for vegetative growth, asexual development, abiotic stress responses, and full virulence in F. pseudograminearum.