The possibility that differences in stomatal conductance between upper and lower surfaces of amphistomatous leaves are adaptations to differences in CO2 exchange characteristics for the two surfaces was investigated. The ratio of upper to lower stomatal conductance was found to change little in response to light and humidity for well-watered sunflower (Helianthus annuus L.) plants. Stressing the plants (V, = -17 bars) and rewatering 1 day before gas exchange measurements reduced upper conductance more severely than lower in both indoor-and outdoor-grown plants, and caused small changes in conductance ratio with light and humidity. A similar pattern was found using outdoor grown sunflower and cocklebur (Xanthium strumarium L.) plants. Calculated intercellular CO2 concentrations for upper and lower surfaces were always close to identical for a particular set of environmental conditions for both sunflower and cocklebur, indicating that no differences in CO2 exchange characteristics exist between the two surfaces. By artificially creating a CO2 gradient across the leaf, the resistance to CO2 diffusion through the mesophyll was estimated and found to be so low that despite possible nonhomogeneity of the mesophyll, differences in CO2 exchange characteristics for the two surfaces are unlikely. It is concluded that differences in conductance between upper and lower stomates are not adaptations to differences in CO2 exchange characteristics. thermal conductivity for leaves (4) make significant temperature gradients across the leaf improbable in most leaves, and although small differences in ambient humidity may exist between adaxial and abaxial surfaces, under reasonably well-stirred conditions these differences are likely to be small. However, Jones and Slatyer (5) reported a higher mesophyll resistance for CO2 entering through the upper stomata than for the lower, and the data of Vaclavik (12) appear to support this conclusion. These data, plus consideration of the anisolateral nature of the mesophyll in many C3 dicotyledonous species, raise the possibility that different CO2 exchange characteristics may exist for CO2 entering through one surface or the other, caused by either differences in resistance to CO2 diffusion through the intercellular spaces or differences in photosynthetic characteristics between palisade and spongy mesophyll cells. Although differences in carbon metabolism between these two types of cells have been shown not to exist (7), differences in electron transport reactions are indicated by differences in fluorescence characteristics between upper and lower surfaces of leaves (1). Photosynthesis and transpiration were determined using a gas exchange system which allowed measurements of upper and lower surfaces independently. A clamp-on type chamber with the leaf forming the barrier between the two chambers was used, and pressure was equalized in the two chambers to prevent gas flow through the leaf. Light was provided by a 300-w cool-beam floodlight, or for later experiments, by a 400-w meta...