Water deficit is the primary constraint of soybean [Glycine max (L.) Merr.] yield, and a physiological understanding of processes affected by water deficit is a key step in identifying and improving drought tolerance in soybean. The objectives of this research were to evaluate biomass and nitrogen accumulation patterns and water use efficiency (WUE) as possible mechanisms associated with the drought tolerance of Jackson. Biomass accumulation of Jackson was contrasted with the PI416937, which also has demonstrated tolerance to drought. For water-deficit treatment, total biomass accumulation was negligible for PI416937, but biomass accumulation continued at approximately 64 % of the well-watered treatment of Jackson. Transpirational losses for Jackson and PI416937 were approximately the same for the water-deficit treatment, indicating that Jackson had superior WUE. Isotopic discrimination of 13 C relative to 12 C also indicated that Jackson had higher WUE. Results indicated that increased WUE for Jackson under water deficit showed it was tolerant to drought rather than had an avoidance mechanism.