The effects of cold acclimation on primary metabolism in actively growing poplar ( Populus tremula L. x P. tremuloides Michaux) were studied. Three-month-old poplar plants were exposed to chilling stress (4 degrees C) and compared to plant material kept at a control temperature (23 degrees C). This treatment did not affect the survival of the plants but growth was almost stopped. The freezing tolerance of the adult leaves increased from -5.7 degrees C for the control plants to -9.8 degrees C after 14 days of exposure to 4 degrees C. During acclimation, the evolution of soluble carbohydrate contents was followed in the leaves. Sucrose, glucose, fructose and trehalose accumulated rapidly under chilling conditions, while raffinose content increased after one week at 4 degrees C. Proteomic analyses, by bidimensional electrophoresis, performed during this stage revealed that a large number of proteins had higher expression, while much less proteins disappeared or had a lower abundance. MALDI-TOF-MS analyses enabled ca. 30 spots to be proposed for candidate proteins. Among the accumulating or appearing proteins proposed, about a third presented similarities with chaperone-like proteins (heat shock proteins, chaperonins). In addition, dehydrins and other late embryogenesis abundant proteins, i.e., stress-responsive proteins, detoxifying enzymes, proteins involved in stress signalling and transduction pathways were also activated or newly synthesised. Finally, cold exposure induced a decrease in the candidate proteins involved in cell wall or energy production.