Abstract: In order to maintain the maternal excellent features in offspring and refrain from difficult rooting in vegetative propagation of hybrid aspen, established a juvenility and container rootinghardening micropropagation system in the present work. The result showed that explants taken from the sprouting between rootshoot junctions of maternal plants, sterilized by soaked 10% effective chlorine of sodium dichloroisocyanurate (NaDCC) solution for 25min, cultured on MS medium, 7591% of them regenerated juvenile shoots. The explants regenerated more than 3 juvenile shoots were selected and subcultured on the same fresh medium in each 30 days, each explant produced 122 shoots on average. The shoots from explants multiplicated one time before did container rooting and hardening. The shoots increased 373 times on MS medium containing 6BA 05mg/L and NAA 02mg/L, respectively. The rooting rate reached the highest at 9889% when IBA was 20mg/L in rooting medium. The container plantlets hardened in reversion and acclimatization by shifting and opening gradually the tissue culture bottles. The container rooted aspen plants survived at 925% in the acclimatization stage.