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Members of the Diatrypaceae are predominantly saprotrophic on the decaying wood of angiosperms worldwide and the family has received little attention due to its difficult taxonomy. However, the recent detection of several pathogenic species, once considered saprotrophic, associated with the wood of diseased grapevines has increased interest in this family. The diversity of tropical species is less well known and more poorly sampled in phylogenetic studies than temperate species. In the present study, we investigated the diversity of diatrypaceous fungi from three areas in the Brazilian semi-arid region and performed phylogenetic analyses of the family based on the entire internal transcribed spacer (ITS) region and partial beta-tubulin gene. Twenty-eight new ITS and 19 new beta-tubulin sequences were generated, representing eight species distributed in five clades. Diatrypella atlantica, Eutypa guttulata, Eutypella cearensis, and Peroneutypa diminutispora are proposed here as new species, while Eutypella microtheca and P. curvispora are new records for Brazil. All eight species are described, illustrated, and discussed.