Comparative Metabolomic Analysis of <i>Botrytis cinerea</i> Treated with Pyrrolnitrin and Phenylpyrrole Fungicides

Pyrrolnitrin, a natural antifungal compound from <i>Burkholderia</i> sp. O33 was compared with the phenylpyrrole fungicides fludioxonil and fenpiclonil through metabolomic analysis of <i>Botrytis cinerea</i>. Phenylpyrrole fungicides are traditionally known to disrupt the high-osmolarity glycerol (HOG) pathway. However, recent studies suggest additional mechanisms for fludioxonil, including reactive oxygen species (ROS) generation and mitochondrial dysfunction. ROS generation and membrane permeability assays showed that pyrrolnitrin induced the highest ROS accumulations, while phenylpyrrole fungicides induced both ROS production and increased membrane permeability. Metabolomic analysis revealed that pyrrolnitrin and phenylpyrrole fungicides inhibit key metabolic pathways, including the TCA cycle, amino acid metabolism, and nucleic acid metabolism, with consistent metabolite changes across these pathways. This study identifies mitochondrial dysfunction and metabolic disruption as shared mechanisms of pyrrolnitrin and phenylpyrrole fungicides.