Based on 2354 bp of mitochondrial DNA (12S rRNA, ND4, cyt b) and 2573 bp of nuclear DNA (C-mos, ODC, R35), we re-examine the phylogenetic relationships of Nilssonia species. Individual and combined analyses of mitochondrial and nuclear DNA using Maximum Likelihood and Bayesian approaches confirm the monophyly of the genus. While mitochondrial data alone could not resolve the phylogenetic position of N. formosa, nuclear data support a sister group relationship of N. formosa and the remaining Nilssonia species. Combined analyses of mitochondrial and nuclear DNA suggest the following branching pattern, with N. formosa as the sister taxon of the remaining species: N. formosa + ((N. gangetica + N. leithii) + (N. hurum + N. nigricans)). Among the samples we studied is the first record of N. formosa for Yunnan, China, and the first record of wild-living N. nigricans for Bangladesh. In N. gangetica, each of the studied major river basins harbours a genetically distinct population, suggesting that at least three distinct management units should be distinguished: (1) Brahmaputra River; (2) Indus and Ganges Rivers plus Ganges Delta; and (3) Mahanadi River.
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