It is know that many plants live in symbiosis with microorganisms that can be found on their interior, the endophytes. Environment and tissue type are modulating factors of this community, in which most of these microorganisms produce important antimicrobial molecules and they may be powerful biocontrol agents in agriculture. Thus, the aim of this study was to evaluate the community of Endophytic fungi from Eremanthus erythropappus in anthropogenic and natural areas (with human interaction, natural habitat and planned planting) of Minas Gerais State, Brazil, through cultivation-based approach and verify their antimicrobial activity against phytopathogenic fungal and pathogenic bacteria. The endophytic fungi isolated were identified by sequencing of the ITS region and subjected an in vitro antagonism test. The antagonisms that show antibiosis were submitted to tests on split plates to verify the volatile compound production. In the pairing testes, the endophytic fungi of the genera Cryptosporiopsis, Diaporthe, Xylaria, Paraconiothyrium and Camarosporium presented antibiosis against phytopathogenic fungi by releasing compounds in the medium. To our knowledge, this paper is the first report on the isolation of twelve genera fungi in E. erythropappus besides verifying their antagonist capacity, which opens the way for discovery of bioactive substances produced by endophytic fungi that inhibit pathogens.