The endophytic fungal biodiversity of unique plants like Peperomia argyreia (Miq.) É. Morren (Piperaceae) has antimicrobial properties and can be employed for infection treatment. Fungal isolates were obtained from appropriately treated plant tissues cultured in solid media, characterized by morphology, and identified by molecular biology using ITS and NL primers. The antimicrobial properties of fungal extracts were analyzed by combining microdilution and bioautographic assays complemented with metabolic profiling by automated thin-layer chromatography and 1H NMR techniques. Thirty-one filamentous fungi were isolated and characterized by ITS and/or D1/D2 region amplification of rDNA, identified as Thermothielavioides, Trichoderma, Cyphellophora, Cladosporium, Arcopilus, Plectosphaerella; Chaetomium, Sporothrix, Alboefibula, and Penicillium. Thermothielavioides spp. inhibited Staphylococcus aureus ATCC 25923; moreover, Penicillium westlingii P4 showed inhibitory activity on Ascochyta rabiei AR2. The bioactivity-guided fractionation of the EtOAc extract (MIC = 62.5 μg/mL) of P. westlingii P4 allowed the purification of citrinin as the main inhibitory compound (MIC = 62.5 μg/mL). Peperomia argyreia harbors a rich and diverse endophytic community able to produce bioactive molecules. Citrinin, with a minor influence of volatile compounds biosynthesized by P. westlingii P4, was responsible for the inhibition of A. rabiei AR2.
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