We describe herein a new species of Vitreorana from the Cerrado biome at Serra da Canastra National Park, Vargem Bonita, Minas Gerais, Southeastern Brazil. Morphological and molecular data from one nuclear gene (proopiomelanocortin) show that the new species belongs to the genus Vitreorana, with a strong support of posterior probability of Bayesian inference. Vitreorana franciscana sp. nov. is characterized by its slender body; snout subovoid in dorsal view; outer tarsal and ulnar ridges with low and small, thickened tubercles; cloacal ornamentation composed of an aggregation of small, thickened tubercles, with two elevations on each side of anal opening forming a slight flap, all highlighted by white pigment; and reticulated arrangement of melanophores on dorsum. Furthermore, we describe the calling repertoire and provide data on natural history of the new species.