Abstract: An in vitro method was developed for the rapid multiplication of Boesenbergia pulcherrima (Wall.) Kuntze, a threatened species of Zingiberaceae. Shoot tip explants developed from rhizome were transferred to different multiplication media containing various concentrations and combinations of BAP, KN and IAA. Though MS medium supplemented with single cytokinin simulated shoot development, showed poor multiplication. Shoot tip explants achieved better response in BAP than in KN. Addition of KN (0.5 mg L-1) to BAP (1.0 mg L-1) containing medium promoted the rate of multiplication and obtained average 5.8 shoots in 5 weeks. Since rooting was also observed along with caulogenesis, the plantlets were straight away transferred to greenhouse conditions. They were successfully established in the greenhouse condition with 85% survival. Hardened plantlets planted in the trial plot exhibited normal regeneration in the subsequent year and developed bulbils. In the following year, 65% of the tissue cultured plants exhibited regeneration. The vegetative and floral characters of the field grown plants were similar to their wild parents. Germination of bulbils produced from tissue cultured plants further confirm the suitability of in vitro plantlets for conservation purpose. The system suggests a feasible method for replenishing the wild population as the in vitro progenies accomplished their life cycle successfully in the field.