Fusarium oxysporum pathogenicity is believed to require the activity of cell wall-degrading enzymes. Production of these enzymes in fungi is subject to carbon catabolite repression, a process that in yeast is mostly controlled by the SNF1 (sucrose non-fermenting 1) gene. To elucidate the role of cell wall-degrading enzymes in F. oxysporum pathogenicity, we cloned and disrupted its SNF1 homologue ( FoSNF1). The fosnf1 mutants had a reduced expression of several genes encoding cell wall-degrading enzymes and grew poorly on certain carbon sources. Infection assays on Arabidopsis thaliana and Brassica oleracea revealed that progression of wilt symptoms in plants infected by fosnf1 mutants was considerably delayed, in comparison with those infected by a wild-type strain. In conclusion, mutations in FoSNF1 prevent F. oxysporum from properly derepressing the production of cell wall-degrading enzymes, compromise the utilization of certain carbon sources, and reduce its virulence on A. thaliana and B. oleracea.