Although the genus Pseudocercospora has a worldwide distribution, it is especially diverse in tropical and subtropical countries. Species of this genus are associated with a wide range of plant species, including several economically relevant hosts. Preliminary studies of cercosporoid fungi from Brazil allocated most taxa to Cercospora, but with the progressive refinement of the taxonomy of cercosporoid fungi, many species were relocated to or described in Pseudocercospora. Initially, species identification relied mostly on morphological features, and thus no cultures were preserved for later phylogenetic comparisons. In this study, a total of 27 Pseudocercospora spp. were collected, cultured, and subjected to a multigene analysis. Four genomic regions (LSU, ITS, tef1 and actA) were amplified and sequenced. A multigene Bayesian analysis was performed on the combined ITS, actA and tef1 sequence alignment. Our results based on DNA phylogeny, integrated with ecology, morphology and cultural characteristics revealed a rich diversity of Pseudocercospora species in Brazil. Twelve taxa were newly described, namely P. aeschynomenicola, P. diplusodonii, P. emmotunicola, P. manihotii, P. perae, P. planaltinensis, P. pothomorphes, P. sennae-multijugae, P. solani-pseudocapsicicola, P. vassobiae, P. wulffiae and P. xylopiae. Additionally, eight epitype specimens were designated, three species newly reported, and several new host records linked to known Pseudocercospora spp.