Abstract: The current year vegetative shoots of ‘Wuci 2’ were used as explants, the effects of different disinfection methods and hormone concentration on the disinfection effect of explants, primary culture, proliferation culture and rooting culture were studied, in order to establish tissue culture system of Rosa roxburghii f.eseiosa Ku.The results showed that the survival rate was 85.28% after the explants were washed with water and disinfected with 75% alcohol for 15 seconds and 0.1% HgCl₂ for 8 minutes.The contaminated plants were washed in ultrasonic washing solution, adding carbendazim 60 mg·L^-1 and penicillin 40 mg·L^-1 in the washing solution, and adding carbendazim 60 mg·L^-1 and penicillin 40 mg·L^-1 in the medium was the best treatment.The suitable medium for the primary culture of ‘Wuci 2’ was MS+1 mg·L^-1 6-BA+0.1 mg·L^-1 NAA,and the germination rate reached 92.04%.The suitable proliferation culture medium was MS+0.2 mg·L^-1 6-BA+0.1 mg·L^-1 NAA,and the proliferation coefficient was 3.9.The rooting rate of ‘Wuci 2’ was 93.17% on the medium of 1/2MS+0.1 mg·L^-1 IBA+20 g·L^-1 sucrose+0.5 g·L^-1 activated carbon.The results of this study were of great significance for the industrial production and conservation of germplasm resources of ‘Wuci 2’.