This work presents a phylogenetic study on the Myiobius genus, a passerine group recognized to Onychorhynchidae family, We estimated the evolutionary relationships among their species, as well as intraspecific analyzes in two of them, it was possible to recover their subespecies relationships, and biogeographic barriers influence, as rivers, in their diversification. For this, two mitochondrial markers (ND2 and COI) and two nuclear loci (ACO1 and βfib5) were used from 90 individuals with a wide distribution in South and Central America. The data confirmed reciprocal monophyly between lineages from west and from east of Andes; We recovered close relationship between M. barbatus / sulphureipygius / villosus species, and evolutionary distance of M. atricaudus; moreover, two of its sympatric and cryptic species (M. barbatus and M. atricaudus) still hybridize to each other, although they have been recovered as evolutionarily distant. It was verified that main Amazon basin rivers influenced the diversification of genus species, recovering populations from different interfluves as distinct evolutionarily from others.
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