The objective of the present investigation was to establish in vitro culture and plant regeneration methods from leaf base and rhizome bud explants of Z. officinale. The MS medium with 1.0 mg/l 2, 4-D proved to be the best for callus induction from leaf base explants. Shoot regeneration was achieved after subculturing the calli in different media formulation and 8.0 mg/l BA with 0.2 mg/l 2, 4-D was found to be the best for multiple shoot regeneration from callus through organogenesis. MS medium supplemented with 5.0 mg/l NAA was the best formulation for successful culture establishment as well as shoot proliferation from rhizome bud explant. Multiple shoot proliferation was noticed at 4th subculture in medium with 5.0 mg/l NAA and shoot proliferation was increased with the increased number of subculture. Activated charcoal (AC) enhanced multiple shoot proliferation and the optimum pH level for shoot formation was 5.5 - 6.0 in the medium. MS + 4.0 mg/l NAA proved to be the best for root induction. Rooted shoots (plantlets) were gradually acclimatized and successfully established in polybag soil.