Discovery and Optimization of MoEno1-Targeting Antifungal Compounds for Rice Blast Control

Developing fungicides with novel modes of action is essential for sustainable rice blast management. Here, we investigated the glycolytic enzyme MoEno1 in Magnaporthe oryzae as a potential antifungal target. Deletion of MoEno1 impaired vegetative growth, melanin production, conidiation, and pathogenicity. Deep-learning-assisted affinity prediction and structure-based virtual screening identified isobavachalcone (IBC) as a MoEno1-binding hit. Structure-guided optimization of IBC yielded 59 derivatives, among which II-24 showed enhanced antifungal activity, stronger MoEno1 binding, and greater inhibition of enolase activity. II-24 treatment was associated with reduced melanin accumulation and acetyl-CoA levels, together with perturbations in energy metabolism. II-24 also showed in planta efficacy, broad-spectrum antifungal activity, low phytotoxicity, low toxicity to zebrafish, predicted low mammalian toxicity, and enhanced activity in combination with tebuconazole. These findings support MoEno1 as a potential antifungal target and II-24 as an early stage lead for rice blast control.