Using simultaneous measurements of leaf gas exchange and chlorophyll fluorescence, we determined the excitation partitioning to photosystem II (PSII), the CO2/O2 specificity of ribulose-1,5-bisphosphate carboxylase/oxygenase, the dark respiration in the light, and the alternative electron transport rate to acceptors other than bisphosphoglycerate, and the transport resistance for CO2 in the mesophyll cells for individual leaves of herbaceous and tree species. The specificity of ribulose-1,5-bisphosphate carboxylase/oxygenase for CO2 was determined from the slope of the O2 dependence of the CO2 compensation point between 1.5 and 21% O2. Its value, on the basis of dissolved CO2 and O2 concentrations at 25.5[deg]C, varied between 86 and 89. Dark respiration in the light, estimated from the difference between the CO2 compensation point and the CO2 photocompensation point, was about 20 to 50% of the respiration rate in the dark. The excitation distribution to PSII was estimated from the extrapolation of the dependence of the PSII quantum yield on F/Fm to F = 0, where F is steady-state and Fm is pulse-satuarated fluorescence, and varied between 0.45 and 0.6. The alternative electron transport rate was found as the difference between the electron transport rates calculated from fluorescence and from gas exchange, and at low CO2 concentrations and 10 to 21% O2, it was 25 to 30% of the maximum electron transport. The calculated mesophyll diffusion resistance accounted for about 20 to 30% of the total mesophyll resistance, which also includes carboxylation resistance. Whole-leaf photosynthesis is limited by gas phase, mesophyll diffusion, and carboxylation resistances in nearly the same proportion in both herbaceous species and trees.