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Onychostoma virgulatum, new species, is described from the Qiupu River, a tributary on the Southern bank of the lower Yangtze River at Shitai County, southern Anhui Province, South China. It shares with O. fusiforme and O. meridionale the presence of a longitudinal dark brown stripe extending along the lateral line, a character separating them from all other congeners of the moderate-mouth group diagnosed by having a slightly arched or nearly transverse mouth opening (with the extremities slightly curved posteriorly), its width being equal to or slightly less than the width of head at this same point, and a short postlabial groove extending along half of the length of the lateral margin of the lower jaw. Onychostoma virgulatum differs from both in the presence of two pairs of barbels in adults, from O. fusiforme in the body depth, caudal-peduncle depth, and position of pelvic and anal fins, and from O. meridionale in the structure of the last simple dorsal-fin ray.