A natural intergeneric hybrid between Rhodeus pseudosericeus and Acheilognathus signifer was collected in their sympatric area, the Jucheongang (stream), the Namhangang (river), Korea. Morphological characters as well as mitochondrial cytochrome b (mt-cyb) gene were investigated to confirm the natural hybrid origin. In the comparison of morphological characters, the natural hybrid appeared to have intermediate characters between two parental species in several key characters such as the band of dorsal fins, the color pattern of body, and the body spot and stripe. Also, the collected specimen had an incomplete lateral line with a large number of pored scales. In analysis of mt-cyb gene, it revealed that the maternal species of the natural hybrid appeared to be R. pseudosericeus due to their 100% sequence identity. Thus, such results gave convincing evidence for the occurrence of a natural intergeneric hybrid between Rhodeus pseudosericeus and Acheilognathus signifer.