In this work, proteomics was used to study the influence of both optimal and low temperatures on growth and development in a vernalization-requiring winter wheat (Triticum aestivum L. cv Cheyenne) after prolonged times of treatment. For this purpose, plants were grown at optimal temperature (20°C) for 14 days (zero point) after which half were transferred to conditioned chambers kept at 4°C for a period of 63 days. Cold tolerance, as estimated from lethal temperatures (LT(50)), and phenological development, as measured by final leaf number (FLN) and shoot apex dissection, were determined. Proteomic analysis indicated a down-accumulation of several photosynthesis-related proteins and a concomitant increase in abundance of some Calvin cycle enzymes. A cold-induced accretion of soluble sugars and proline was observed as well. In parallel, an increase of proteolysis accomplished by an up-modulation of TCA cycle enzymes was also noticed, probably suggesting an efficient recycling of amino acids as energy source. Proteomic analysis of plants grown at optimal temperature allowed to specifically discriminate cold-induced proteins and highlight molecular processes driven by vernalization. Among identified proteins typically involved in vernalization responses and floral transition we observed a marked increase of wrab17, wcor18 and glycine-rich RNA-binding proteins.