Single node cuttings with one mature leaf were taken from Rosa ×hybrida `Baroness' and rooted in water culture. The plants were subjected to either 90% (high) or 70% (moderate) relative humidity (RH) in climate chambers. Single stem roses with intact roots were transferred to 40% (low) RH to investigate the stomatal response to water stress. Moderate RH plants showed decreasing leaf conductance from day 1 to day 3 during both light and dark phases, in contrast to high RH roses, which showed almost similar leaf conductances during the 3 days. Leaf samples were studied with a light microscope (LM) and a scanning electron microscope (SEM) to quantify morphological and structural changes. Epidermal imprints showed a significantly higher number of stomata and longer stomata, as well as a wider stomatal apertures on roses grown at high RH. The high RH leaves showed a reduced density of vascular tissue and thinner leaves when compared to moderate RH leaves. Enlarged intercellular air-space (ICA) was found due to a reduced number of spongy and palisade mesophyll cells. No obvious difference in shape, size, undulation or the structure of the epicuticular wax was observed in SEM between high and moderate RH grown leaves. In conclusion, roses subjected to high RH showed differences in leaf anatomy, stomatal morphology and stomatal function, which may explain the loss of water control of these plants. Stomatal ontogenesis should occur at RH conditions below 85% to secure roses with a high postharvest quality potential.