VdPex30, a peroxisomal membrane protein, is involved in carbon metabolism, stress response, and pathogenicity of <i>Verticillium dahliae</i>

ABSTRACT The fungus Verticillium dahliae causes Verticillium wilt in plants and has a broad host range. The peroxisomal membrane protein gene VdPex30 (VDAG_09094), identified in a host-induced gene silencing screen, plays a crucial role in fungal pathogenicity. VdPex30 encodes a 671-amino-acid protein of the Pex24p subfamily, with expression strongly induced during early infection. VdPex30 - RNA interference transgenic tobacco showed enhanced resistance to V. dahliae . Δ VdPex30 mutant strains displayed impaired growth, sporulation, and tolerance to oxidative, cell wall, and osmotic stresses, along with reduced virulence on cotton, and all these defects were restored by complementation. The ΔVdPex30 strains were hypersensitive to H ₂ O ₂ , accumulated higher reactive oxygen species, and downregulated oxidative response-related genes. Transcriptome analysis identified 3,623 differentially expressed genes (DEGs), confirmed by reverse transcription quantitative PCR (RT-qPCR), and a subset of representative DEGs was further validated by RT-qPCR. Moreover, VdMIase (VDAG_01341) was identified as a novel VdPex30-interacting protein. These findings reveal that VdPex30 is vital for growth, stress adaptation, and pathogenicity of V. dahliae , providing a potential target for Verticillium wilt control. IMPORTANCE Verticillium dahliae causes Verticillium wilt, a destructive vascular disease that severely threatens many crops, including cotton. Identifying fungal genes required for infection can enable more durable control strategies. Here, we demonstrate that VdPex30, a peroxisomal membrane protein identified in a host-induced gene silencing screen, is essential for fungal growth, stress adaptation, and pathogenicity. VdPex30 knockout (Δ VdPex30 ) strains showed impaired carbon utilization and sporulation, hypersensitivity to oxidative stress, elevated intracellular reactive oxygen species, and markedly reduced virulence on cotton, all of which were restored by genetic complementation. Moreover, plant-mediated RNA interference targeting the fungal VdPex30 transcript enhanced host resistance to infection. Together, our findings reveal a critical link between peroxisome function and fungal virulence and highlight VdPex30 as a promising target for RNA interference-based management of Verticillium wilt.