A high frequency plant regeneration pro-tocol via direct organogenesis has been develo-ped for Punica granatum L. cv. Kandhari Kabuli, a medicinally important horticultural tree. Cotyle-don explants procured from 15-20-days-old in vitro grown seedlings were used as explants. MS medium supplemented with 2.0 mg/l 6-benzylaminopurine (BAP) and 1.5 mg/l naphtha-lene acetic acid (NAA) was optimal for shoot re-generation, which resulted into highest frequen-cy of shoot regeneration (67.89%) along with maximum average number of shoots formed per explant (5.38) and average shoot length (3.62 cm). Shoot multiplication and elongation took place on the same medium. Half strength MS medium containing 0.1 mg/l NAA and 0.02% ac-tivated charcoal was found most effective for rooting of shoots resulting into 80.12% root re-generation frequency. Regenerated plantlets were transferred to plastic cups containing auto-claved peat moss and successfully acclimatized with 85% survival rate. This is the first report on in vitro plant regeneration through direct orga-nogenesis in P. granatum L. cv. Kandhari Kabuli using cotyledon explants.