Selecting drought-tolerant and more water-efficient wheat genotypes is a research priority, specifically in regions with irregular rainfall or areas where climate change is expected to result in reduced water availability. The objective of this work was to use high-throughput measurements with morphophysiological traits to characterize wheat genotypes in relation to water stress. Field experiments were conducted from May to September 2018 and 2019, using a sprinkler bar irrigation system to control water availability to eighteen wheat genotypes: BRS 254; BRS 264; CPAC 01019; CPAC 01047; CPAC 07258; CPAC 08318; CPAC 9110; BRS 394 (irrigated biotypes), and Aliança; BR 18_Terena; BRS 404; MGS Brilhante; PF 020037; PF 020062; PF 120337; PF 100368; PF 080492; and TBIO Sintonia (rainfed biotypes). The water regimes varied from 22 to 100% of the crop evapotranspiration replacement. Water stress negatively affected gas exchange, vegetation indices, and grain yield. High throughput variables TCARI, NDVI, OSAVI, SAVI, PRI, NDRE, and GNDVI had higher yield and morphophysiological measurement correlations. The drought resistance index indicated that genotypes Aliança, BRS 254, BRS 404, CPAC 01019, PF 020062, and PF 080492 were more drought tolerant.