We quantified a broad range of Arabidopsis thaliana (Col-0) leaf phenotypes for initially warm-grown (25/20 degrees C day/night) plants that were exposed to cold (5 degrees C) for periods of a few hours to 45 d before being transferred back to the warm, where leaves were allowed to mature. This allowed us to address the following questions: (1) For how long do warm-grown plants have to experience cold before developing leaves become irreversibly cold acclimated? (2) To what extent is the de-acclimation process associated with changes in leaf anatomy and physiology? We show that leaves that experience cold for extended periods during early development exhibit little plasticity in either photosynthesis or respiration, and they do not revert to a warm-associated carbohydrate profile. The eventual expansion rate in the warm was inversely related to the duration of previous cold treatment. Moreover, cold exposure of immature/developing leaves for as little as 5 d resulted in irreversible changes in the morphology of leaves that subsequently matured in the warm, with 15 d cold being sufficient for a permanent alteration of leaf anatomy. Collectively, these results highlight the impact of transitory cold during early leaf development in determining the eventual phenotype of leaves that mature in the warm.