Abstract: The tissue culture regeneration system of kiwifruit 'Hort16A' (Actinidia chinensis 'Hort16A') introduced from abroad was established. The callus formation rate and adventitious bud differentiation rate were investigated using theL₉(3⁴)orthogonal experiment, proliferation of adventitious buds were further studied using single factor randomized block design of experiment, and the rooting of kiwifruit was researched using theL₉(3⁴)orthogonal experiment. The result indicated that the highest callus formation rate (97.78%±1.92%) was investigated from the explants in the MS medium contained 0.05 mg/L NAA and 0.3 mg/L ZT after culturing 20 d. That the differentiation rate and height of adventitious buds were (84.44%±1.92%) and (1.52±0.29) cm respectively were the best differentiation effect surveyed on the explants in this medium after another 14 d culturing. The biggest proliferation rate (3.67±0.33) and height (2.93±0.12) cm of adventitious buds were surveyed on the buds in the MS medium contained 1.5 mg/L ZT after culturing 30 d. The best rooting that rooting ratio, root length, root width and number of roots generated per shoot were (98.9%±1.9%), (7.60±0.44) cm, (0.47±0.07) cm and (7.67±2.19) respectively was investigated from the shoots in the 1/2 MS medium contained 0.7 mg/L IBA and 0.1 g/L activated carbon.