More frequent droughts and an increased pressure on water resources, combined with social licence to operate, will inevitably decrease water resources available for fully irrigated cotton production. Therefore, the long-term future of the cotton industry will require more drought tolerant varieties that can perform well when grown in rainfed cropping regions often exposed to intermittent drought. A trait that limits transpiration (TRLim) under an increased vapour pressure deficit (VPD) may increase crop yield in drier atmospheric conditions and potentially conserve soil water to support crop growth later in the growing season. However, this trait has not been tested or identified in cotton production systems. This study tested the hypotheses that (1) genetic variability to the TRLim VPD trait exists among ten genotypes in the Australian cotton breeding program; (2) genotypes with a TRLim VPD trait use less water in high VPD environments, and (3) that variation in yield responses of cot