The photosynthetic characterization of Populus euphratica and their response to increasing groundwater depth and temperature were analyzed based on net photosynthetic rate (PN), stomatal conductance (gs), intercellular CO2 concentration (Ci), transpiration rate (E), water use efficiency (WUE) and stomatal limitation (Ls) measured by a portable gas-exchange system (LI-6400) in the lower reaches of the Tarim River. Light-response curves were constructed to obtain light-compensation and light-saturation points (LCP and LSP), maximum photosynthetic rates (Pmax), quantum yields (AQY), and dark respiration rates (RD). The growth condition of P. euphratica, soil moisture, and groundwater depth in the plots were analyzed by field investigation. The results showed that the growth condition and photosynthetic characterization of P. euphratica were closely related to groundwater depth. The rational groundwater depth for the normal growth and photosynthesis was 3-5 m, the stress groundwater depth for mild drought was more than 5 m, for moderate drought was more than 6 m, for severe drought was more than 7 m. However, P. euphratica could keep normal growth through a strong drought resistance depended on the stomatal limitation and osmotic adjustment when it faced mild or moderate drought stress, respectively, at a normal temperature (25°C). High temperature (40°C) significantly reduced PN and drought stress exacerbated the damage of high temperature to the photosynthesis. Moreover, P. euphratica would prioritize the resistance of high temperature when it encountered the interaction between heat shock and water deficit through the stomata open unequally to improve the transpiration of leaves to dissipate overheating at the cost of low WUE, and then resist water stress through the osmotic adjustment or the stomatal limitation. and H. H. Zhou ... [et al.].