Abstract: Inter-specific hybrids of Oncorhynchus mykiss(♀) and Oncorhynchus masou masou(♂) were produced in this study．The results showed that the fertilization rate was 90.00％, eyed egg rate was 80.52%, the hatching rate was 90.68％ and the survival rate was 30.68％ at fry stage．The genetic analysis on the relationship between the F₁ progenies and their parents were carried out using microsatellite markers．The results are as follows: (1) Three pairs of primers only gave good images in Oncorhynchus mykiss. Ten pairs of primers gave clear bands in Oncorhynchus mykiss and Oncorhynchus masou masou, six showed specific bands and four showed no specific bands between the parents. (2) Genetic differentiation between the parents was significant at these microsatellite loci. Hybridization between Oncorhynchus mykiss and Oncorhynchus masou masou was feasible. The inherited character between parents and hybrid follows the Mendelian inheritance rule and they are indeed hybrids in genetic material. (3) The hybrid F₁ has a relative genetic similarity of 0.461 7 and 0.596 5 from Oncorhynchus mykiss and Oncorhynchus masou masou, separately. The hybrid F₁ has a genetic distance of 0.772 9 and 0.516 7 from Oncorhynchus mykiss and Oncorhynchus masou masou, separately. The former is farther than the later, which elucidates that the hybrid F₁ has an unequal heredity difference to its parents and a genetic inclination to its paternity. This is also supported by UPGMA. The results of the research will be useful for the further study on the hybridization between Oncorhynchus mykiss and Oncorhynchus masou masou.