Multi-gene phylogeny of the primary freshwater crab genus Ptychophallus Smalley, 1964 (Pseudothelphusidae: Ptychophallinae) from the Neotropical region.

The diversity of primary freshwater crabs of Central America is underestimated and poorly known, specially when considering both Atlantic and Pacific drainages distribution. Among the family Pseudothelphusidae Ortmann, 1893, the genus Ptychophallus Smalley, 1964 was recently revised using morphological data and encompassing 14 valid species, which are distributed exclusively in Costa Rica and Panama. Despite the informative scenario constructed with this previous study, some uncertainties remained such as those related to the populations distributed along different drainages and the lack of complete information regarding the phylogenetic relationships among the species that belong to this genus. Our phylogenetic reconstruction based on multigenes revealed four lineages. Ptychophallus costaricensis, P. colombianus, P. lavallensis, P. montanus, and P. tumimanus are well-established species, while P. paraxanthusi, P. exilipes and P. tristani were recovered as non-monophyletic in some relationships. Ptychophallus tristani and P. paraxanthusi were determined to be close relatives one with other, but with some representatives showing no clear correspondence and positioning in the molecular phylogeny, which raised doubts about the validity as separate species and/or the accuracy of identifications for some GenBank sequences. Ptychophallus tumimanus and P. montanus are morphologically very similar, but they are positioned in distinct clades. The genetic results confirmed a previous proposal that P. barbillaensis is a junior synonym of P. uncinatus, and P. bilobatus is closely related to P. uncinatus. The findings revealed the presence of one peculiar lineage with one species from Costa Rica compound by the resurrected P. campylus and another species from Costa Rica, originally identified as "P. cf. exilipes"; its morphology, however, did not match with any other analyzed species, which requires further analysis. Finally, possible misidentification in some sequences deposited in the GenBank were detected and should be reviewed. The reconstructed molecular phylogeny for the genus that inhabits both Atlantic and Pacific drainages of Central America provides a better understanding of the knowledge on the evolution of freshwater crabs and represents key information that will serve as baseline for further taxonomic studies on the cryptic biodiversity of this group.