Elevated atmospheric carbon dioxide (Ca) usually reduces stomatal conductance, but the effects on plant transpiration in the field are not well understood. Using constant-power sap flow gauges, we measured transpiration from Quercus myrtifolia Willd., the dominant species of the Florida scrub-oak ecosystem, which had been exposed in situ to elevated Ca (350 µmol mol−1 above ambient) in open-top chambers since May 1996. Elevated Ca reduced average transpiration per unit leaf area by 37%, 48% and 49% in March, May and October 2000, respectively. Temporarily reversing the Ca treatments showed that at least part of the reduction in transpiration was an immediate, reversible response to elevated Ca. However, there was also an apparent indirect effect of Ca on transpiration: when transpiration in all plants was measured under common Ca, transpiration in elevated Ca-grown plants was lower than that in plants grown in normal ambient Ca. Results from measurements of stomatal conductance (gs), leaf area index (LAI), canopy light interception and correlation between light and gs indicated that the direct, reversible Ca effect on transpiration was due to changes in gs caused by Ca, and the indirect effect was caused mainly by greater self-shading resulting from enhanced LAI, not from stomatal acclimation. By reducing light penetration through the canopy, the enhanced self-shading at elevated Ca decreased stomatal conductance and transpiration of leaves at the middle and bottom of canopy. This self-shading mechanism is likely to be important in ecosystems where LAI increases in response to elevated Ca.