Because atmospheric carbon dioxide is the ultimate source of all land-plant carbon, workers have suggested that pCO(2) level may exert control over the amount of C-13 incorporated into plant tissues. However, experiments growing plants under elevated pCO(2) in both chamber and field settings, as well as meta-analyses of ecological and agricultural data, have yielded a wide range of estimates for the effect of pCO(2) on the net isotopic discrimination (Delta delta C-13(p)) between plant tissue (delta C-13(p)) and atmospheric CO2 (delta C-13(CO2)). Because plant stomata respond sensitively to plant water status and simultaneously alter the concentration of pCO(2) inside the plant (c(i)) relative to outside the plant (c(a)), any experiment that lacks environmental control over water availability across treatments could result in additional isotopic variation sufficient to mask or cancel the direct influence of pCO(2) on Delta delta C-13(p). We present new data from plant growth chambers featuring enhanced dynamic stabilization of moisture availability and relative humidity, in addition to providing constant light, nutrient, delta C-13(CO2), and pCO(2) level for up to four weeks of plant growth. Within these chambers, we grew a total of 191 C-3 plants (128 Raphanus sativus plants and 63 Arabidopsis thaliana) across fifteen levels of pCO(2) ranging from 370 to 4200 ppm. Three types of plant tissue were harvested and analyzed for carbon isotope value: above-ground tissues, below-ground tissues, and leaf-extracted nC(31)-alkanes. We observed strong hyperbolic correlations (R >= 0.94) between the pCO(2) level and Delta delta C-13(p) for each type of plant tissue analyzed; furthermore the linear relationships previously suggested by experiments across small (10-350 ppm) changes in pCO(2) (e. g., 300-310 ppm or 350-700 ppm) closely agree with the amount of fractionation per ppm increase in pCO(2) calculated from our hyperbolic relationship. In this way, our work is consistent with, and provides a unifying relationship for, previous work on carbon isotopes in C-3 plants at elevated pCO(2). The values for Delta delta C-13(p) we determined in our ambient pCO(2) chambers are consistent with the Delta delta C-13(p) values measured in large modern datasets of plants growing within the Earth's wettest environments, suggesting that it may be possible to reconstruct changes in paleo-pCO(2) level from plants that grew in consistently wet environments, if delta C-13(CO2) value and initial pCO(2) level can be independently quantified. Several implications arise for the reconstruction of water availability and water-use efficiency in both ancient and recent plant Delta delta C-13(p) values across periods of changing pCO(2) level. For example, the change in Delta delta C-13(p) implied by our relationship for the rise in pCO(2) concentration observed since 1980 is of the same magnitude (= similar to 0.7 parts per thousand) as the isotopic correction for changes in delta C-13(CO2) required by the input of C-13-depleted carbon to the atmosphere. For these reasons, only the portion of the terrestrial isotopic excursion that persists after accounting for changes in pCO(2) concentration should be used for the interpretation of a change in paleo-environmental conditions. (C) 2012 Elsevier Ltd. All rights reserved.