Identification of Aspartate Aminotransferase FocAST2 as a Novel Target of Albendazole in <i>Fusarium oxysporum</i> f. sp. <i>cubense</i> TR4

Fusarium wilt, caused by <i>Fusarium oxysporum</i> f. sp. <i>cubense</i> Tropical Race 4 (<i>Foc</i> TR4), poses a severe threat to global banana production. While fungicides remain the most cost-effective strategy for disease control, the emergence of drug resistance underscores the urgent need for novel molecular targets. In this study, we identified albendazole (ABZ), a benzimidazole derivative, as a broad-spectrum antifungal agent that effectively suppresses the growth of <i>Foc</i> TR4 by targeting a key metabolic enzyme. Through RNA-seq, drug affinity responsive target stability (DARTS), and microscale thermophoresis (MST) assays, we demonstrated that ABZ directly binds to and inhibits aspartate aminotransferase FocAST2. Functional analysis revealed that the loss of <i>FocAST2</i> severely impairs conidiation, increases sensitivity to oxidative stress, and disrupts infection-related morphogenesis. Notably, <i>FocAST2</i>-deficient mutants exhibit phenotypic alterations similar to those induced by ABZ treatment, further supporting its role as a key mediator of ABZ activity. These findings establish FocAST2 as a direct target of ABZ and a promising candidate for the development of novel fungicides, providing mechanistic insights into fungal metabolism and fungicide resistance management.