Crown rot caused by Fusarium pseudograminearum remains a significant global challenge to wheat production. In this study, the role of FpCBS in F. pseudograminearum were investigated by creating deletion mutants and assessing their impacts on hydrogen sulfide (H ₂S) production, oxidative stress adaptation, and pathogenicity, supported by transcriptomic and metabolomic analyses. Deletion of FpCBS resulted in a significant reduction in H ₂S production, accompanied by increased levels of homocysteine and cysteine, suggesting the importance of FpCBS for H ₂S biosynthesis. Furthermore, Δ Fpcbs mutants displayed diminished pathogenicity and decreased resistance to H ₂O ₂. Also, Δ Fpcbs mutants correlated with reduced catalase activity. Transcriptomic data revealed altered gene expression related to oxidative stress and virulence, including genes encoding cell wall-degrading enzymes, mycotoxins, and polyketides. Altogether, we proposed that F. pseudograminearum uses CBS-dependent H ₂S production to facilitate host infection, adaptation to host-induced oxidative stress, and synthesis of virulence factors. These findings highlight the potential of FpCBS as a novel target for developing strategies to control wheat crown rot.