Cichlid fishes are an important model system in evolutionary biology, primarily because of their exceptional diversity. However, while some cichlid assemblages, such as the ones of the African Great Lakes, have received considerable attention, others are not well studied, including many riverine species. Here, we focus on theAustraloheros autranispecies group and first report a new record ofAustraloherosin the upper Paranaíba River drainage, extending the known distribution range of this genus. Through Bayesian inference and maximum likelihood phylogenetic analyses of the mitochondrial cytochromebgene of these specimens as well as available sequences, we assigned the newly discovered population toAustraloheros barbosae. We corroborate the monophyly of theA. autranispecies group and the presence of three species in the upper/middle Paraíba do Sul River basin as well as molecular diagnostic characters for each. Finally, we provide evidence for a recent expansion ofA. barbosae.Supplementary InformationThe online version contains supplementary material available at 10.1007/s10228-022-00888-9.