Chromosome banding patterns obtained by ammoniacal silver staining (Ag-AS) and alkaline Giemsa (CBG) have been analysed in several amphibian species of the genus Odontophrynus from South America. Ag-AS bands were found at secondary constrictions, mainly of chromosomes 4 and 11. The CBG technique revealed centromeric and telomeric constitutive heterochromatin on almost all chromosomes of these species. Moreover, intercalary bands were found at particular sites of several chromosomes. Some inter- and intra-population polymorphisms were found for the Ag-AS and C-banding patterns. The species variability in the number and position of the Ag-AS bands, known to be regions of active ribosomal cistrons, as well as the specific sites of intercalary heterochromatin, are used to discuss the possible evolutionary relationships among these species.