Gray mold, caused by the fungus Botrytis cinerea, is a key postharvest disease of strawberries, resulting in substantial fruit rot and economic losses. This study elucidated the comprehensive antifungal mechanism of punicalagin against B. cinerea. The antifungal action of punicalagin primarily involved the disruption of fungal cell membrane integrity, as evidenced by the induction of cytoplasmic leakage, accumulation of malondialdehyde and reduction in ergosterol and trehalose. Ultrastructural analysis revealed severe hyphal damage after punicalagin treatment. Punicalagin also induced apoptosis through mitochondrial membrane dysfunction and the accumulation of reactive oxygen species (ROS). Furthermore, transcriptomic analysis identified 2589 downregulated and 1157 upregulated genes in punicalagin-treated mycelia, with a particular impact on membrane lipid and carbohydrate metabolism pathways. These transcriptomic alterations, representing impaired energy production and antioxidant systems, were further validated by corresponding metabolomic analysis. Additionally, punicalagin treatment preserved the quality of strawberry fruit by improving cell wall structure to maintain firmness and augmenting phenolic content, potentially enhancing resistance. These multifaceted mechanisms collectively contributed to the exceptional efficacy of punicalagin in controlling B. cinerea. These findings suggest that punicalagin could be a promising natural antifungal alternative, offering effective control of gray mold and maintaining the quality of strawberries in storage.
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