The aim of this study was to evaluate gas exchange, efficiency of the photosynthetic apparatus, and antioxidant activity in Copaifera langsdorffii Desf. The seedlings were cultivated under different conditions of water availability, in order to improve the utilization efficiency of available water resources. The seedlings were cultivated in four different water retention capacities (WRC-25%, 50%, 75%, and 100%), and evaluated at four different time (T-30, 60, 90, and 120 days). During the experimental period, seedlings presented the highest values for carboxylation efficiency of Rubisco (A/Ci), intrinsic water use efficiency (IWUE = A/gs), chlorophyll index, and stomatal opening, when grown in the substrate with 75% WRC, but the stomatal index (SI) was less the 25% WRC. The efficiency of photosystem II was not significantly altered by the treatments. Comparison between the extreme treatments in terms of water availability, represented by 25% and 100% WRC, represent stress conditions for the species. Water availability causes a high activity of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase) in the plant.