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.