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

Fusarium wilt, caused by Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc 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 Foc 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 FocAST2 severely impairs conidiation, increases sensitivity to oxidative stress, and disrupts infection-related morphogenesis. Notably, FocAST2-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.