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Recently, <i>Diaporthe</i> has been considered the most frequently isolated genera of endophytic fungi, having a broad spectrum of host plants and a worldwide distribution. The endophytic <i>Diaporthe</i> strain used in the present work came from the Fungal Collection of Phytopathology and Mycology Subdepartment, University of Life Sciences in Lublin (Poland), and was isolated from healthy <i>Prunus domestica</i> shoots during previous studies. Due to the possibility of using the <i>Diaporthe</i> endophytes as a promising option for plant disease management, the main goal of the research was to study the antagonistic effect of endophytic <i>Diaporthe</i> strain against six phytopathogens: <i>Verticillium dahliae</i>, <i>Botrytis cinerea</i>, <i>Fusarium avenaceum</i>, <i>F. sprotrichioides</i>, <i>Alternaria alternata</i>, and <i>Trichothecium roseum</i> based on the dual culture assay and to determine the catabolic profile of the endophyte by using Biolog FF Plates. The dual-culture test assay revealed the ability of the endophytic <i>Diaporthe</i> to limit the growth of all tested pathogens. The growth inhibition percentage ranged from 20% (<i>V. dahliae</i>) to 40% (<i>T. roseum</i>). A distinct zone of inhibition occurred between the endophytic <i>Diaporthe</i> and the pathogens <i>T. roseum, V. dahliae</i>, and <i>B. cinerea</i> in the co-growth combinations. As for the catabolic profile results, the most intensive utilization of carbon substrates was observed after 168 h of incubation. The growth of the analyzed strain was observed on 79 media containing carbohydrates, carboxylic acids, amino acids, amines and amides, polymers, and others. The most effective decomposition was observed in the polymers group, the least in amines and amides. Molecular identification indicated that this strain was closely related to the <i>Diaporthe eres</i> species complex.