Abstract: We used leaves (12-d-old) of Rosa hybrida as explants and MS medium to examine the effect of combinations and concentrations of growth regulators on callus, adventitious buds and root induction. The optimal medium for inducing calli was MS medium with 3.0 mg·L^–12,4-D+1.0 mg·L^–16-BA; 100% callus induction ratio was obtained. 6-BA played a key role in adventitious bud induction. Adventitious buds were regenerated from calli on MS medium containing 1.0 mg·L^–16-BA, (0.05–0.5) mg·L^–1NAA and (0.02–0.2) mg·L^–1TDZ. The callus redifferentiation ratio was about 92.6% in MS medium supplied with 1.0 mg·L^–16-BA+0.1 mg·L^–1NAA+0.02 mg·L^–1TDZ. Auxin is an important factor for inducing rooting; 100% rooting was obtained on 1/4 MS medium containing 0.1 mg·L^–1NAA. We established a tissue culture and high-frequency regeneration system of R. hybrida.