Abstract: Odontomachus chelifer are ants widely distributed in the Brazilian cerrado that belong to the group of the poneromorphs. The species is polygynous (more than one functional queen per nest) and participates in several important interactions such as seed dispersal. It is known that O. chelifer are susceptible to habitat fragmentation, which is capable of affecting the genetic diversity in their nests. In ants, the genetic variation of the colonies is directly influenced by polygyny, which can be mediated by environmental factors such as scarcity of resources. Thus, it is important to understand the potential effects of habitat fragmentation on polygyny and genetic structure of O. chelifer in order to conserve their ecological populations and roles. For this purpose, in this work we developed microsatellite markers for the species to investigate their genetic diversity and polygynous habit, as well as their possible associations with border effects in the Cerrado Ver menos