A wide range of soybean cultivars is available on the market and understanding the physiological response and yield of these materials is fundamental to develop new management systems. Thus, the objective of the present study was to assess ecophysiological parameters and yield of soybean cultivars under field conditions. The experiment was carried out on a farm located in the municipality of Açailândia, Maranhão, Brazil. Three commercial cultivars were used (SC1, SC2 and SC3), and gas exchanges, SPAD index, Fv/Fm, photosynthesis index (PI), instantaneous water use efficiency (WUE) and intrinsic instantaneous of the use of water (iWUE) were assessed during the vegetative (V5) and reproductive (R5) stages. In addition, the biomass and production components were obtained. A randomized complete block design was used, with three cultivars and six replications. SC2 obtained the best mean for the photochemical variables. SC2 was more efficient at both development stages in WUE, but the maximum iWUE values were obtained in SC3. The SC2 cultivar obtained the best responses in the main variables analyzed, resulting in a higher yield.