We used DNA sequence data to test the morphology-based taxonomy and examine the biogeography of the Brazilian Atlantic Rain Forest genus Promitobates. Most species are well differentiated morphologically, and a previous morphological phylogeny recovered the genus as monophyletic. However, some of these species have overlapping geographical distributions and considerable intraspecific variation, perhaps representing a species complex. Mitochondrial (12S rRNA and COI) and nuclear (ITS2 and 28S) genes were sequenced from 132 specimens collected from 27 localities. The results are consistent with significant mitochondrial introgression among the species P. ornatus, P. hatschbachi, P. lager, P. bellus and P. intermedius (the ‘P. ornatus species complex’), with one specimen identified as a hybrid between P. nigripes and this complex. A phylogeographic study of the complex was conducted using mitochondrial haplotypes. This revealed remarkably poor dispersal among populations, with only one case of a shared haplotype, and very low genetic diversity. The phylogeny showed a clear break between populations on either side of a narrow region of forest, suggesting an important historical event separated these lineages in the genus. The analyses also pointed to population breaks that date back several millions of years or extremely small effective population sizes, depending on the mutation rate.