The response of mesophyll conductance to nitrogen and water availability differs between wheat genotypes

•Increased mesophyll conductance may improve crop productivity and water-use efficiency.•The response of mesophyll conductance to water and nitrogen availability differed between wheat genotypes.•Leaf intrinsic water-use efficiency was more closely related to stomatal than to mesophyll conductance for wheat.•The limitation to photosynthesis due to mesophyll conductance increased when wheat plants were droughted. Increased mesophyll conductance (gm) has been suggested as a target for selection for high productivity and high water-use efficiency in crop plants, and genotypic variability in gm has been reported in several important crop species. However, effective selection requires an understanding of how gm varies with growth conditions, to ensure that the ranking of genotypes is consistent across environments. We assessed the genotypic variability in gm and other leaf gas exchange traits, as well as growth and biomass allocation for six wheat genotypes under different water and nitrogen availabilities. The wheat genotypes differed in their response of gm to growth conditions, resulting in genotypic differences in the mesophyll limitation to photosynthesis and a significant increase in the mesophyll limitation to photosynthesis under drought. In this experiment, leaf intrinsic water-use efficiency was more closely related to stomatal conductance than to mesophyll conductance, and stomatal limitation to photosynthesis increased more in some genotypes than in others in response to drought. Screening for gm should be carried out under a range of growth conditions.