Evidence is presented which suggests that the mesophyll cell walls of cotton leaves may influence observed rates of transpiration.The net diffusive flux of water vapour, from the upper and lower surfaces of a leaf, was compared with the flux of nitrous oxide through a leaf and evidence obtained of an extra resistance in the water-vapour pathway associated with water transport in the mesophyll cell walls.This extra resistance appeared to be insignificant at low transpiration rates and in turgid leaves, but increased with transpiration rate and dehydration. The most likely explanation for its origin appeared to be a reduction in hydraulic conductivity across the internal cuticle which lines the outer surfaces of the mesophyll cell walls. In turn this served to reduce the relative vapour pressure at the sites of evaporation.The experiments were conducted under conditions where stomatal opening was induced by CO2-free air. Under normal conditions stomatal closure would tend to reduce the development of this extra resistance. Even so, the results throw doubt on the validity of the long-standing assumption that the water-vapour pressure at the evaporation sites is equal to the saturation vapour pressure under all conditions.