The subfamily Potamotrygoninae, the only extant clade of elasmobranchs exclusive to freshwater environments, encompasses four genera and 38 species distributed across almost every major South American river basin. Despite their importance in the ornamental fish trade, the taxonomy and evolutionary relationships within potamotrygonines have not yet been resolved. Here, we present a comprehensive molecular phylogeny for the Neotropical freshwater stingrays, based on extensive species and population sampling (35 species and > 350 individuals from drainages across South America). Our phylogeny corroborates the monophyly of the genera Paratrygon and Heliotrygon and the monophyly of the Potamotrygon + Plesiotrygon clade. Within the Potamotrygon + Plesiotrygon clade, we identify a core Potamotrygon clade characterized by short branches, low nodal support and incongruence with current species-level taxonomy. In the core Potamotrygon clade, specimens of widespread species, such as Potamotrygon motoro and Potamotrygon orbignyi, do not form monophyletic lineages; instead, specimens from these species are often closely related to those of other species from the same river basins. These patterns could be caused by inaccurate taxonomy, hybridization, incomplete lineage sorting and rapid diversification. We discuss the conservation of Neotropical freshwater stingrays from a phylogenetic perspective and suggest ways to prioritize potamotrygonid conservation efforts with respect to endemism and evolutionary distinctiveness.

Evolutionary transitions between marine and freshwater ecosystems have occurred repeatedly throughout the phylogenetic history of fishes. The theory of ecological opportunity predicts that lineages that colonize species-poor regions will have greater potential for phenotypic diversification than lineages invading species-rich regions. Thus, transitions between marine and freshwaters may promote phenotypic diversification in trans-marine/freshwater fish clades. We used phylogenetic comparative methods to analyze body size data in nine major fish clades that have crossed the marine/freshwater boundary. We explored how habitat transitions, ecological opportunity, and community interactions influenced patterns of phenotypic diversity. Our analyses indicated that transitions between marine and freshwater habitats did not drive body size evolution, and there are few differences in body size between marine and freshwater lineages. We found that body size disparity in freshwater lineages is not correlated with the number of independent transitions to freshwaters. We found a positive correlation between body size disparity and overall species richness of a given area, and a negative correlation between body size disparity and diversity of closely related species. Our results indicate that the diversity of incumbent freshwater species does not restrict phenotypic diversification, but the diversity of closely related taxa can limit body size diversification. Ecological opportunity arising from colonization of novel habitats does not seem to have a major effect in the trajectory of body size evolution in trans-marine/freshwater clades. Moreover, competition with closely related taxa in freshwaters has a greater effect than competition with distantly related incumbent species.

The various configurations of the jaws, anterior branchial arches and neurocranium provide some of the fundamental synapomorphies, distinguishing the major lineages of sharks, rays and ratfishes. For the Neotropical freshwater stingrays (family Potamotrygonidae), putatively unique skeletal elements, the angular cartilages, are intermediate between the hyomandibula and the lower jaw. These skeletal elements have been proposed as a synapomorphy for the potamotrygonids, but confidence in this character has been limited by poor sampling and taxonomic uncertainty, particularly regarding their ontogeny and homology. The morphology of the angular cartilages across the family is described, and the nomenclature of these structures is standardized. In addition, developmental and functional hypotheses for the origins of the angular cartilages are proposed. These cartilages, formed within the hyomandibular–Meckelian ligament, are suggested to be fibrocartilaginous in origin and aid in jaw kinesis. The angular cartilages are corroborated as a synapomorphy for Potamotrygonidae, although there is great variation within the family and similar structures have arisen independently in other extinct and extant batoid lineages. In potamotrygonines, the angular cartilages appear to have been lost in piscivorous Heliotrygon and Paratrygon, while additional angular cartilages were gained, typically one at a time, in some lineages of Potamotrygon.