Nitrogen fertilization induces dramatic changes in the growth and development of plants, including forest trees. In this study we examined short-term responses of hybrid poplar, Populus balsamifera ssp. trichocarpa x deltoides, to N fertilization. Glasshouse-grown saplings subjected to limiting, intermediate, and luxuriant levels of ammonium nitrate over a 28 d time course demonstrated rapid changes to whole-plant architecture and biomass accumulation. Nitrogen-associated shifts in allocation occurred in temporally distinct stages. Nitrogen availability modulated parameters that affect carbon gain, including light-saturated net photosynthesis and leaf area. These parameters were affected by N-induced changes to leaf maturation and senescence. Leaf area was also affected by N-induced sylleptic branch development. Genes encoding vegetative storage proteins and starch biosynthetic enzymes exhibited contrasting patterns of expression under differential N availability. A gene encoding a previously uncharacterized putative pectin methylesterase inhibitor displayed expression patterns comparable to the starch biosynthetic genes. The results of this study illustrate the phenotypic plasticity that P. balsamifera ssp. trichocarpa x deltoides exhibits in response to differential N availability.