Regulation of ABC Transporters and Ergosterol Biosynthesis by the Transcription Factor FvADS-1 Controls Azole Resistance and Virulence in Fusarium verticillioides

<i>Fusarium verticillioides</i> is a significant agricultural pathogen and an emerging causative agent of invasive fusariosis in clinical settings. <i>Fusarium</i> species frequently exhibit resistance to available antifungal agents, yet the molecular mechanisms underlying azole resistance remain poorly characterized. In this study, we identified the Zn(II)₂Cys₆ transcription factor FvADS-1 as a positive regulator of the azole stress response in <i>F. verticillioides</i>. The transcription of <i>FvADS-1</i> was significantly induced by ketoconazole (KTC), and its deletion increased susceptibility to multiple azole compounds. Mechanistically, FvADS-1 positively regulates the KTC-induced expression of genes encoding ABC transporters and ergosterol biosynthesis enzymes, thereby modulating intracellular KTC accumulation and sterol homeostasis under azole stress. Furthermore, FvADS-1 positively regulates the transcriptional response of peroxisomal genes and contributes to fungal tolerance to oxidative stress. Notably, deletion of <i>FvADS-1</i> attenuates the virulence of <i>F. verticillioides</i> on maize. The function of ADS-1 is evolutionarily conserved: heterologous expression of <i>N. crassa ads-1</i> restored azole resistance in <i>FvADS-1</i> deletion mutant, and the deletion of the <i>F. oxysporum</i> homolog <i>FoADS-1</i> similarly increased azole susceptibility. Collectively, our study demonstrates that the conserved transcription factor ADS-1 plays a central role in regulating azole resistance and virulence in the pathogen <i>F. verticillioides</i>, offering new insights into antifungal resistance mechanisms in pathogenic filamentous fungi.