A phylogenetic analysis using fragments of two nuclear and two mitochondrial genes strongly supported sister group relationships between a new species and , corroborated by unique colour patterns of adult specimens and juveniles, and morphology of the autopalatine bone. The new species is distinguished from its closest congener, , by details of the colouration and number of branchiostegal rays. Both the new species and are endemic to the Rio Paraíba do Sul basin, in the Atlantic Forest of south-eastern Brazil, but occur in distant and disjunct areas. The new species, herein described, is endemic to the upper section of the Rio Grande drainage, a right tributary of the lower Rio Paraíba do Sul, an area situated in the Órgãos Mountain Range. is endemic to an area about 200 km across, in the upper section of the Rio Preto that drains the Itatiaia Massif. However, both species are only known from localities above 1100 m asl, suggesting that they cannot survive in ecological conditions at lower altitudes. This study indicates that efforts are necessary to provide more accurate data on the species diversity and distribution of in the biologically diverse and endangered Atlantic Forest of south-eastern Brazil.