ABSTRCT: Peach is one of the most important fruit crops in the world, belongs to family rosaceae."Nemaguard" peach (Prunus persica L. × Prunus davidiana) is used extensively as rootstock for peach cultivars. The rootstock and the grafted cultivar influence the vegetative and generative mass and the profitability of fruit production moreover in the efficiency of pest and disease management programs and fruit yield. This investigation was carried out to optimize invitro technique for micropropagation of Nemaguard rootstock. Effect of sodium hypochlorite concentrations (0.37, 0.50, 0.75, 1.50, 2.25 and 3.0%) and exposure periods (2, 4, 6 min) on success of sterilization of nemagurd peach explants was examined. Shoots which were sterilized with 3% NaOCl for 6 min, recorded the highest number of sterilized explants (9.7). Efect of cytokinin types and concentration on proliferation of Nemaguard peach was studied MS medium which supplemented with 2 mg/l BAP gave the highest shoot number (9.0 shoots/explant). While, MS medium supplemented with 2 mg/l ADSO4 in combination with 0.5 mg/l IAA resulted in shoot number (3.0 Shoots/explant), which clear that it is lower than the shoot proliferation in presence of BAP. The lowest shoot proliferation was obtained when MS medium supplemented with KIN. Concerning shoot length of nemagurd peach, all types of used cytokinin adversely affected shoot length; while IAA was promote shoot length; MS medium supplemented with 5 mg/l ADSO4 in combination with 0.5 mg/l IAA resulted in the highest shoot length (3.7 cm). Leaves number was affected by types and concentrations of cytokinins. The highest leaves number was possessed when MS medium supplemented with either 5 mg/l KIN in combination with 0.25 mg/l IAA (22 leaves / shoot) or 5 mg/l ADSO4 in combination with 0.5 mg/l IAA (22 leaves / shoot ). The produced shoots were successfully rooted (64%) when transplanted on half strength MS medium supplemented 0.5 mg/l IAA. The rooted plantlets showed 85% successes in acclimatization. Generally, it could be concluded that cytokinin types and concentration affected shoot proliferation of nemagurd peach. Through tested cytokinin, BAP is the most suitable cytokinin for multiplication of nemagurd peach.