Abstract: The young branches of rabbit eye blueberry ‘Tifblue’ were used as experimental materials, the effects of fungicides, medium and growth hormone on explant sterilization, cluster bud induction, subculture propagation, rooting and transplantation of tissue culture seedlings were studied by tissue culture method with different concentrations and types of agents, culture media and auxin, in order to provide reference for the construction of comprehensive and efficient rapid propagation system of blueberry.The results showed that the best sterilization method for explants was 10% NaClO solution disinfection for 15 minutes, the pollution was reduced to 17.34%,and the number of germination was 9.71.The optimal medium for inducing tufted buds in blueberry stem segment was 1/2 WPM+1.5 mg·L^-1 ZT+0.2 mg·L^-1 IBA,and the success rate was 68.36%.The optimal subculture medium was 1/2 WPM+0.1 mg·L^-1 NAA+1.5 mg·L^-1 ZT,the multiplication rate was 5.53,and the average plant reached 5.15 cm.The optimal rooting medium for tissue culture was 1/2 WPM+0.5 mg·L^-1 IBA+200 mg·L^-1 activated carbon（AC）,and the rooting rate, effective rooting rate, root length and number were 87.16%,87.16%,4.39 cm and 6.32,respectively.Moss was the best substrate for tissue culture and the survival rate was 93.47%.