The subfamily Xylocopinae has been recognized as the most basal lineage within the family Apidae, comprising four tribes; Allodapini, Ceratinini, Xylocopini and Manueliini. Relationships among the tribes are not well resolved with morphological data. In particular, Manueliini and Xylocopini have each been placed as the most basal lineage in separate analyses of the subfamily. While relationships within each tribe, excepting Manueliini, have been investigated using molecular data, these data have not been applied to examine the relationships among tribes, which remain controversial. Here we present results of molecular phylogenetic analyses using sequences of CoI, Cytb and EF-1aF1 from members of the four tribes of Xylocopinae. We used available data from other studies in combination with data generated for the three species of Manueliini. Competing phylogenetic hypotheses regarding the alternate positions proposed to Manueliini and Xylocopini were evaluated through statistical tests. The basal position of either Manueliini or Xylocopini has contrasting implications on the evolutionary history of nest architecture, which mediates the potential for contact between adult and immature individuals. Our results indicate that Manueliini is the most basal lineage of Xylocopinae, in agreement with an evolutionary transition from nests having completely sealed cells to nests lacking cells. A nest structure with closed cells prevents physical interactions between adult and immature stages, whereas an open structure provides the opportunity for interactions that may play an important role in the emergence of sociality.