Abstract: In recent years, the Eucalyptus dunnii had become an important afforestation tree species in southern China because of its fast growing and high cold-tolerance. But its seeds mainly depended on imports and the price is expensive, so to carry on the regeneration system of the research was of great significance to solve the problem of seedling. In this paper, a regeneration system of E. dunnii using seed and axillary bud as explants was established, the results showed: Using aseptic sowing technique of E. dunnii, cutting their hypocotyls and cotyledons and cultured in vitro, establishment of adventitious buds and plant regeneration of embryogenesis. The best culture medium was 1/2MS+6-BA0.5+NAA0.2+IBA0.5, the subculture cycle was 25 days, the subculture coefficiency reached 3.1. Furthermore, E. dunnii axillary buds were used to study organogenesis approach. The results showed that 1/2MS+6-BA0.5+NAA0.2+IBA0.5 medium also suitable for axillary buds induction and differentiation. Subculture cycle and propagation coefficient is consistent with the embryogenesis way. The axillary shoots were transferred to rooting medium 1/2MS+NAA0.2+IBA0.5 which come from embryogenesis and organogenesis pathway, 7 days later, successions generated roots, the rooting rate could reach to 70%-71% after 25 days. The results provided the theoretical and practical basis for the rapid propagation and factory production of E. dunnii.