It was generally assumed that the accumulation of vegetative storage protein (VSP) in poplar trees and/or temperate hardwoods did not occur in spring. To test this assumption, the accumulation of the 32-kDa VSP and the differential expression of a gene encoding for the protein in poplars were investigated using light and electron microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDSPAGE) and reverse transcription-polymerase chain reaction (RT-PCR). We report, for the first time, that poplar trees initiated VSP accumulation in new shoots during the development of new shoots in spring under conditions of high temperature and long days. The amount of 32-kDa VSP increased gradually in the stem of new shoots and in two-year-old branches, but there were no detectable changes in its abundance in the bark tissues of trunks during new shoot development. Based on the presence of a 286-bp DNA fragment that is identical to the VSP gene bspA, encoding for the 32-kDa VSP in Populus deltoids Bartr. ex Marsh., the differential expression of the 32-kDa VSP gene in P. canadensis Moench was revealed by RT-PCR. The results indicated that the 32-kDa VSP gene was expressed strongly in new shoots, relative weakly in twoyear-old branches and was not expressed in the trunk during new shoot development. This pattern of VSP accumulation and VSP gene space-time differential expression may be an important mechanism by which stored nitrogen compounds are used preferentially to exogenously available nitrogen and, in addition, the dynamic pattern may also have a role in the regulation of nitrogen metabolism, especially nitrogen uptake by the roots.