An efficient protocol for shoot organogenesis and plant regeneration has been developed for Punica granatum L. cv. Kandhari Kabuli using cotyledon derived callus. Calli were induced from cotyledon explants procured from in vitro germinated seedlings on MS medium supplemented with various concentrations and combinations of plant growth regulators. Optimal callus formation (93.71 %) from cotyledon explants was obtained on MS medium supplemented with 4.0 mg/l BAP and 3.0 mg/l NAA. The highest shoot induction response (72.34 %) was recorded on MS medium containing 2.0 mg/l BAP and 2.0 mg/l NAA leading to maximum average no. of shoots per callus clump (7.16) and shoot length (1.98 cm). The highest percentage of root regeneration (72.50 %) was observed on MS medium supplemented with 0.10 mg/l NAA. The in vitro raised plantlets were successfully acclimatized and grew well in the greenhouse. All the regenerated plantlets appeared normal with respect to morphology and growth characteristics with 80 % survival rate.