Stevia rebaudiana is a medicinally important plant, free-calorie with sweet test and beneficial for diabetic patient. In order to optimization of the micropropagation of this plant, three experiments were conducted by factorial experiment based on completely randomized design (CRD) with three replications. Shoot tip explants were cultured on MS media supplemented with Kin (0, 1 & 2 mg/l) + IAA (0, 0.5, 1 & 1.5 mg/l) (test 1), Kin (3 & 4 mg/l) with IAA (0.5, 1 & 1.5 mg/l) (test 2) and BAP (0, 1, 2, 3 & 4 mg/l) + IBA (0, 0.5, 1 & 1.5 mg/l) (test 3). The shoot number increased with increasing concentrations of Kin in media and in second experiment, the highest shoot number was produced in MS medium supplemented with 4 mg/l Kin + 0.5 mg/l IAA (with an average of 7 shoots per explant). However, the best result of micropropagation was obtained from MS medium containing 2 mg/l BAP + 1.5 mg/l IBA in test 3 (ith an average of 9 shoots per explant). Also, increasing BAP concentrations from 2 mg/l resulted to a significant reduction in shoot number and shoot length and quality of plantlets as well as increase in callusing. In addition, shoot number was dramatically increased by incubation the cultured explants in culture room for two months. The maximum rooting of developed shootlets was achieved from MS medium supplemented with1 mg/l IBA. The rooted plantlets were hardened in culture room and with high successfully established in soil.