Cytogenetic studies and hybridization experiment using two populations of Oryzias latipes from Korea were conducted in order to examine their systematic positions. In the chromosome number of specimens examined. O. sinensis had 46 with a pair of large metacentrics and O. latipes had 48 chromosomes without a pair of large metacentrics. However, the diploid chromosome number of the hybrid between O. latipes and O. sinensis was 47 and they had a large metarcentric chromosome in their karyotype. In the $F_2$ hybrids, distribution of chromosome number was variable among individuals. Hatching rates showed little difference between control and hybrids. However, hatchiabilties of $F_2$ hybrids were decreased by 83.3% as compared with $F_1$ offspring. Incidence of abnormal larvae was increased by 15.3% as compared with their parents. These results indicate that the two karyotype populations formerly regarded as two subspecies would be two distinct allopatric sibling species O. sinensis, and O. latipes.