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Previous work has shown that stomata of growth chamber-grown Vicia faba leaves have an enhanced CO2 response when compared with stomata of greenhouse-grown plants. This guard cell response to CO2 acclimatizes to the environmental conditions on the transfer of plants between the two environments. In the present study, air relative humidity is identified as a key environmental factor mediating the changes in stomatal sensitivity to CO2. In the greenhouse environment, elevation of relative humidity to growth chamber levels resulted in an enhanced CO2 response, whereas a reduction in the light level to that comparable to growth chamber conditions had no effect on stomatal CO2 sensitivity. The transfer of plants between humidified and normal greenhouse conditions resulted in an acclimation response with a time-course matching that previously obtained in transfers of plants between greenhouse and growth chamber environments. The high stomatal sensitivity to CO2 of growth chamber-grown plants could be reduced by lowering growth chamber relative humidity and then restored with its characteristic acclimation time-course by an elevation of relative humidity. Leaf temperature was unchanged during this restoration, eliminating it as a primary factor in the acclimation response. Humidity regulation of stomatal CO2 sensitivity could function as a signal for leaves inside dense foliage canopies, promoting stomatal opening under low light, low CO2 conditions.

Relative humidity is a key factor in the acclimation of the stomatal response to CO2