The Cambeva variegata group (CVG) is endemic to a region situated in the intersection of two endangered biodiversity hotspots, Cerrado and Atlantic Forest, and drained by two important South American river basins, the upper Rio Paraná and upper Rio São Francisco basins. Presently, CVG comprises two nominal species, besides some still undescribed. We first performed a molecular phylogenetic analysis (total of 3368 bp) for five species of the CVG and 30 outgroups, which supported the monophyly of the CVG and its inclusion in Cambeva. Most morphological character states distinguishing the CVG from congeners are also present in Scleronema, possibly consisting of plesiomorphic features. We also performed the first time-calibrated phylogeny of the group, which supported possible relationships between present geographical distribution patterns and palaeogeographical events. The estimated time of origin of CVG in the Middle Miocene is nearly contemporaneous to a past hydrographical configuration when part of the upper Rio Paraná basin was connected to the Rio São Francisco basin. The first CVG lineage split occurring in the Miocene end corresponds to a major break in that palaeo basin. Species diversification between the Pliocene and early Pleistocene is compatible with final drainage rearrangement. This study highlights the urgent need for more detailed studies on the diversity and phylogenetic relationships of still poorly known organisms in this highly diverse and threatened region.
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