Evolution of Sri Lanka's Giant Danios (Teleostei: Cyprinidae: Devario): Teasing apart species in a recent diversification.

The small, colourful freshwater fishes of the cyprinid genus Devario are among the many vertebrate groups that appear to have diversified on Sri Lanka, a continental Indian Ocean island, which is part of the Western Ghats-Sri Lanka Biodiversity Hotspot. Despite Sri Lanka having been connected with India via a wide isthmus intermittently until the Plio-Pleistocene and almost continuously since then, during sea-level low-stands, the number of species of Devario on Sri Lanka is comparable with that on the Indian Peninsula, some 25 times its size. Here, from a sampling of 27 Devario populations across Sri Lanka's major river basins and climatic zones, we present and analyze a phylogeny based on mitochondrial and nuclear DNA data. We show that five species of Devario occur on the island, all but one of which are endemic: Devario malabaricus is widespread throughout the lowlands and parts of central hills of Sri Lanka and southern India. A new narrow-range endemic, here described as D. memorialis sp. nov., was discovered in a remnant rainforest habitat at Aranayake (Ma Oya basin) in this study. It is immediately distinguished from Sri Lankan congeners by having only 8 (vs 9-12) branched dorsal-fin rays and by uncorrected pairwise genetic distances of more than 4.0% and 7.8% for cytochrome c oxidase subunit 1 and cytochrome b, respectively. Our results provide strong support for the monophyly of the entire Sri Lankan diversification of Devario. Divergence-timing analysis suggests the common ancestor dispersed to the island in the late Miocene, with the insular diversification in the island's south-western wet zone taking place during the Plio-pleistocene. There are signs of gene flow between the Indian and Sri Lankan populations of D. malabaricus until the late Pleistocene. Phylogenetic and haplotype-network analyses suggest basin-centric phylogeographic structure within the endemic species; D. malabaricus, however, shows little such structure in the island. Molecular and morphological analyses failed to identify D. annnataliae and D. udenii confidently as distinct species: they are considered synonyms of D. micronema. The morphological variation observed within D. micronema is likely attributable to polymorphism. The discordance between the mitochondrial and nuclear phylogenies for some samples of Devario in the present study suggest signs of mitochondrial introgression.