Long-term exposure of plants to elevated partial pressures of CO 2 (pCO 2 ) often depresses photosynthetic capacity. The mechanistic basis for this photosynthetic acclimation may involve accumulation of carbohydrate and may be promoted by nutrient limitation. However, our current knowledge is inadequate for making reliable predictions concerning the onset and extent of acclimation. Many studies have sought to investigate the effects of N supply but the methodologies used generally do not allow separation of the direct effects of limited N availability from those caused by a N dilution effect due to accelerated growth at elevated pCO 2 . To dissociate these interactions, wheat (Triticum aestivum L.) was grown hydroponically and N was added in direct proportion to plant growth. Photosynthesis did not acclimate to elevated pCO 2 even when growth was restricted by a low-N relative addition rate. Ribulose-1, 5-bisphosphate carboxylase/oxygenase activity and quantity were maintained, there was no evidence for triose phosphate limitation of photosynthesis, and tissue N content remained within the range recorded for healthy wheat plants. In contrast, wheat grown in sand culture with N supplied at a fixed concentration suffered photosynthetic acclimation at elevated pCO 2 in a low-N treatment. This was accompanied by a significant reduction in the quantity of active ribulose-1, 5-bisphosphate carboxylase/oxygenase and leaf N content.Growth at elevated pCO 2 frequently brings about change in plant physiology that is commonly interpreted as acclimation (Drake et al., 1997). Photosynthesis is inextricably involved because CO 2 is the substrate in C 3 species that is limiting at the current atmospheric pCO 2 . However, results from investigations on the effects of elevated pCO 2 on photosynthesis have been inconsistent. The stimulatory response brought about when pCO 2 is suddenly increased (Long, 1991) has often been found to decline with increasing duration of exposure (for review, see Gunderson and Wullschleger, 1994;Sage, 1994; Drake et al., 1997), but some experiments have failed to find any long-term effect, either in controlled environments (Radoglou and Jarvis, 1990;Wong, 1990) or in the field (Arp and Drake, 1991;Jones et al., 1995;Pinter et al., 1996). Why, then, is the acclimatory response so varied? Species differences can no doubt account for some of the variability, but often the same species in apparently similar conditions can yield different results with different investigators (Sage, 1994). This fact in itself suggests that there may be some uncontrolled factor(s) in the experimental design that may be crucial to the acclimatory response of photosynthesis.Evidence that additional factors may be interacting with the CO 2 response was brought to prominence by Arp (1991), who, after reviewing the data from several investigations using a variety of experimental designs, suggested that root restriction by pot size had a significant effect on the acclimatory response. Limited rooting volume was suggested to creat...