Accumulation of recently photosynthesized sucrose in the guard-cell wall is the empirical foundation for a hypothesis that links the rates of photosynthesis, translocation, and transpiration (Plant Physiology 114, 109–118). Critical assumptions of this hypothesis were tested by use of Vicia faba, an apoplastic phloem loader. Following measurements of the leaflet-apoplastic-water volume (by P–V isotherm analysis) and the guard-cell wall volume (by 3-D analysis), intact leaflets were fed dilute solutions of mannitol, an impermeant non-toxic osmolyte. Even at bulk-leaflet mannitol concentrations that would have only a negligible osmotic effect on stomata, transpiration at constant temperature, water-vapour pressure, air movement and irradiance was diminished up to 25%, compared with controls. This effect on transpiration, a manifestation of smaller stomatal aperture size, was explained by accumulation of mannitol, up to 350 mol m−3, in the estimated aqueous volume of the guard-cell wall. The conclusion is that mannitol, a xenobiotic with structural similarity to sucrose, can move throughout the apoplast of a transpiring leaflet and accumulate in an osmotically significant concentration in the guard-cell wall. These data therefore provide support for a new role for sucrose as a signal metabolite that integrates essential functions of the whole leaf. In addition, the results raise questions about the physiological or experimental accumulation of other guard-cell-targeted apoplastic solutes such as plant growth regulators, particularly abscisic acid, and ions.