We report global gene expression patterns of poplar in response to chilling stress. A total of 1,085 significantly differentially expressed genes, involved in photosynthesis, signal transduction, and regulation of transcription, were identified. To understand the gene network underlying the response to chilling stress in the poplar, Populus simonii, we determined the genome transcript expression profile using an Affymetrix GeneChip with 56,000 genes. Our results revealed 11,626 cold-responsive genes, with 5,267 upregulated and 6,359 downregulated. In terms of biological processes, gene ontology (GO) analysis indicated that cold-induced genes were enriched in response to temperature stimulus, reactive oxygen species, and hormone stimulus. GO terms including cellular nitrogen compound metabolic processes, photosynthesis, and generation of precursor metabolites and energy were enriched in the cold-repressed genes. The functional annotation of differentially expressed genes revealed genes involved in photosynthesis, calcium/calmodulin-mediated signal transduction, abscisic acid (ABA) homeostasis and transport, and antioxidant defense systems. Gene expression analysis showed that the majority of genes involved in photosynthesis were repressed, but the THF1 gene was induced, suggesting that it may play an important role in the production of vesicles for leaf development under low-temperature conditions. Several genes involved in calcium/calmodulin-mediated signal transduction, ABA homeostasis and transport, and antioxidant defense systems were significantly induced under chilling stress, suggesting that they may act as positive regulators in the enhanced low-temperature tolerance of poplar. Several transcription factors had divergent expression patterns, suggesting they have variable functional responses to abiotic stress. This profile of global gene expression patterns during chilling stress will be valuable for future studies on the molecular mechanisms of chilling tolerance in woody plants.