Evaluation of the Effects of Epicoccum nigrum on the Olive Fungal Pathogens Verticillium dahliae and Colletotrichum acutatum by 1H NMR-Based Metabolic Profiling

Olive trees are a cornerstone of Mediterranean agriculture but face significant threats from diseases such as Verticillium wilt and olive anthracnose. These diseases, caused by Verticillium dahliae and Colletotrichum spp., respectively, result in significant economic losses and degrade olive oil quality. While traditional chemical treatments present environmental risk, sustainable alternatives such as biological control agents (BCAs) are gaining attention. Epicoccum nigrum, an antagonistic fungus, has shown potential as a BCA due to its production of antimicrobial secondary metabolites. This study aimed to observe whether E. nigrum has an antagonistic ability against V. dahliae and C. acutatum, and to elucidate the metabolic interactions between these fungi using NMR-based metabolomics. E. nigrum showed inhibitory effects on the growth of C. acutatum and V. dahlia of 44.97% and 38.73% respectively. Metabolomic profiling revealed distinct biochemical responses in E. nigrum, V. dahliae, and C. acutatum under mono- and dual-culture. Multivariate statistical analysis highlighted the metabolic shifts in mycelia and identified the primary metabolites, such as glutamine, 4-aminobutyrate, and phenylalanine that are involved in adaption for survival in stress conditions such as the presence of a competitor. The results could be important for a better understanding of the primary fungal metabolism, which is still poorly characterized. Further investigation is needed, but these results suggest that E. nigrum could serve as a BCA, offering a more sustainable approach to managing olive diseases.