Experiments were performed to determine if growth at elevated partial pressure of CO2 altered the sensitivity of leaf water vapour conductance and rate of CO2 assimilation to the leaf-to-air difference in the partial pressure of water vapour (Δw). Comparisons were made between plants grown and measured at 350 and 700 μPa Pa−1 partial pressures of CO2 for amaranth, soybean and sunflower grown in controlled environment chambers, soybean grown outdoors in pots, and orchard grass grown in field plots. In amaranth, soybean and orchard grass, both the absolute and the relative sensitivity of conductance to Δw at the leaf surface were less in plants grown and measured at the elevated CO2. In sunflower, there was no change in the sensitivity of conductance to Δw for the two CO2 partial pressures. Tests in soybeans and amaranth showed that the change in sensitivity resulted from elevated CO2 during the measurement of the Δw response. Assimilation rate of CO2 was not altered by Δw in amaranth, which has C4 metabolism. In sunflower, the assimilation rate of plants grown and measured at elevated CO2 was insensitive to Δw, consistent with the response of assimilation rate to intercellular CO2 partial pressure in the prevailing range. In soybean, the sensitivity of assimilation rate to Δw was not different between CO2 treatments, in contrast to what would be expected from the response of assimilation rate to intercellular CO2 partial pressure.