Abstract: In this report, tissue culture and regeneration system from leaves in vitro have been extensively investigated utilizing Populus Leucopyramidalis as experiment materials. The results showed that the anti-oxidation materials polyvinylpyrrolidone (PVP) and ascorbic acid (Vc) decreased browning efficiently; the best medium for proliferation was 1/2 MS+ BA0.3mg/L + NAA 0.05mg/L with differentiation percentage of 87% and proliferation multiples of 7.5 respectively, and there was a great significant difference compared with other treatments. The differentiation and regeneration of adventitious shoot from leaves increased significantly in the medium MS containing BA0.3mg/L and NAA0.05mg/L supplemented with 12mg/L sodium nitroprusside (SNP) in which regeneration frequency and average number of shoots were 100% and 12 respectively. The best rooting medium was 1/2 MS+ IBA0.3mg/L .