Populus species are important resources for industry and in scientific study on biological and agricultural systems. Our objective was to enhance the frequency of plant regeneration in Himalayan poplar(Populus ciliata wall. ex Royle). The effect of TDZ alone and in combination with adenine and NAA was studied on the regeneration potential of petiole explants. The explants were excised from Himalayan poplar plants grown in glasshouses. After surface sterilization the explants were cultured on shoot induction medium. High percentage shoot regeneration(86 %) was recorded on MS medium supplemented with 0.004 mg L~-1TDZ and 79.7 mg L~-1adenine. The regenerated shoots for elongation and multiplication were transferred to MS + 0.5 mg L~-1BAP + 0.2 mg L~-1IAA +0.3 mg L-1GA3. Root regeneration from shoots developed in vitro was observed on MS medium supplemented with 0.10 mg L~-1IBA. Himalayan poplar plantlets could be produced within2 months after acclimatization in a sterile mixture of sand and soil. We developed a high efficiency plant regeneration protocol from petiole explants of P. ciliata. Populus species are important resources for industry and in scientific study on biological and agricultural systems. Our objective was to enhance the frequency of plant regeneration in Himalayan poplar (Populus ciliata wall. Ex Royle). The effect of TDZ alone and in combination with adenine and NAA was studied on the regeneration potential of petiole explants. The explants were excised from Himalayan poplar plants grown in glasshouses. High surface shoot the explants were cultured on shoot induction medium. High percentage shoot regeneration (86%) was recorded on MS medium supplemented with 0.004 mg L ~ -1TDZ and 79.7 mg L ~ -1adenine. The regenerated shoots for elongation and multiplication were transferred to MS + 0.5 mg L ~ -1 BAP + 0.2 mg L ~ -1 IAA +0.3 mg L ~ 1GA3. developed in vitro was observed on MS medium supplemented with 0.10 mg L ~ -1 IBA. Himalayan poplar plantlets could be produced within 2 months after acclimatization in a sterile mixture of sand a nd soil. We developed a high efficiency plant regeneration protocol from petiole explants of P. ciliata.