The stingless bee Tetragonisca angustula Latreille 1811 is one of the most widespread bee species in the Neotropics, distributed from Mexico to Argentina. However, this wide distribution contrasts with the low distance that females travel to build new nests whereas nothing is known about male dispersion. Previous studies of T. angustula were ambiguous concerning its genetic structure and were based only on nuclear markers and on small and/or limited sample size. Here we evaluate the genetic structure of several populations of T. angustula by using mitochondrial DNA and microsatellites. These markers can help us to detect differences in the migratory behavior between males and females. Our results showed that the populations were highly differentiated suggesting that both females and males are low dispersers. Therefore, its continental distribution might consist of several different taxa.