Mesophyll CO2 conductance and leakiness are not responsive to short- and long-term soil water limitations in the C4 plant Sorghum bicolor.

Breeding economically important C4 crops for enhanced whole-plant water-use efficiency (WUEplant ) is needed for sustainable agriculture. WUEplant is a complex trait and an efficient phenotyping method that reports on components of WUEplant , such as intrinsic water-use efficiency (WUEi , the rate of leaf CO2 assimilation relative to water loss via stomatal conductance), is needed. In C4 plants, theoretical models suggest that leaf carbon isotope composition (δ13 C), when the efficiency of the CO2 -concentrating mechanism (leakiness, ϕ) remains constant, can be used to screen for WUEi . The limited information about how ϕ responds to water limitations confines the application of δ13 C for WUEi screening of C4 crops. The current research aimed to test the response of ϕ to short- or long-term moderate water limitations, and the relationship of δ13 C with WUEi and WUEplant , by addressing potential mesophyll CO2 conductance (gm ) and biochemical limitations in the C4 plant Sorghum bicolor. We demonstrate that gm and ϕ are not responsive to short- or long-term water limitations. Additionally, δ13 C was not correlated with gas-exchange estimates of WUEi under short- and long-term water limitations, but showed a significant negative relationship with WUEplant . The observed association between the δ13 C and WUEplant suggests an intrinsic link of δ13 C with WUEi in this C4 plant, and can potentially be used as a screening tool for WUEplant in sorghum.