This study was conducted to identify the taxonomic differences in the characteristics of Rhodeus ocellatus and Acheilognathus yamatsutae during their early life history via an interspecific hybridization experiment. Results showed that the hatching rates of the cross-bred eggs were 60% for cross OY (R. ocellatus ♀×A. yamatsutae ♂) and 70% for cross YO (A. yamatsutae ♀×R. ocellatus ♂). The dorsal fin rays of YO were similar to those of R. ocellatus. The hybridization test of A. yamatsutae and R. ocellatus living in the same water stream revealed that no interspecies reproductive isolation occurred. OY and YO showed similar maternal and paternal characteristic. The juvenile of OY did not resemble both paternal and maternal fin rays, indicating a difference. The hatching larvae of the hybrids showed the morphological characteristics of their parents but varied in terms of the shapes of their parts.