A protocol of direct organogenesis from unexpanded leaflets of four Rosa rugosa (Rosa rugosa Thunb.) cultivars was developed.Effects of explants,induction period under dark condition,combinations of plant growth regulators were investigated.The results showed that the highest frequency of adventitious buds induction and plant regeneration were obtained by using a three-stage procedure:initially excised unexpanded leaflets were incubated on half-strength MS (1/2 MS) medium supplemented with 0.05 mg/L NAA and certain concentration TDZ and BA adjusted for each cultivar under the dark condition for 10-15 d and subsequently transferred into light condition for 20 d,adventitious buds excised from the leaves were finally transferred to the MS medium with 0.5 mg/L BA,0.01mg/L NAA in the light for elongation.Bud formation capacity was also significantly affected by the genotype.Among the four cultivars,the highest shoot regeneration response (70.6%) was obtained for ‘Tang White’.For cultivar ‘Puce Dragon’,there were significant effects on adventitious bud induction when incorporating of 1.0 mg/L BA and 0.3 mg/LTDZ on the basic medium.For cultivar ‘Tang White’,‘Puce Dragon’ and ‘Purple Branch’,keeping the cultures under darkness for 10 d promoted shoot induction of leaflets explants,while for ‘Tang Red’,the best results were obtained in darkness for 15 d.Elongated shoots were excised and rooted best on 1/2 MS medium supplemented with 0.5 mg/L IBA.Rooted plantlets,more than 4 roots and longer than 1 cm,were successfully transplanted to soil for acclimatization.