Mass Spectrometry‐Based Molecular Networks Applied to Study the Molecular Interaction Between Two Endophytic Fungi Isolated From <i>Tibouchina granulosa</i> (Vell.) Cogn (Melastomataceae)

Establishing a microorganism as an endophyte involves complex molecular interactions with its host plant and a broader microbial community. Precise detection methods and comprehensive metabolite annotation are essential to study these interactions. This study focused on characterizing the chemical composition of metabolites produced by two endophytic fungi, Colletotrichum siamense and Xylaria berteroi, isolated from Tibouchina granulosa leaves in axenic conditions and coculture. We examined the fungal metabolites using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HR-MS/MS) and analysis tools like Cytoscape and Global Natural Products Social Molecular Networking. Co-cultivation revealed unique compounds not produced in isolation, including N-acetyltryptamine. A total of 32 compounds were identified, many with biotechnological potential due to their bioactivities. The untargeted metabolomics approach demonstrated that interactions among these T. granulosa endophytes can activate inactive metabolic pathways under axenic conditions, potentially producing novel bioactive molecules. This study is the first study of the chemical profile and interaction between endophytes isolated from T. granulosa.