This study explored reductions in tissue nitrogen concentration ([N]) at elevated CO2 concentrations ([CO2]), and changes in plant water and N uptake. Eucalyptus saligna Sm. seedlings were grown under three [CO2] levels (preindustrial (280 μL L–1), current (400 μL L–1) or projected (640 μL L–1)) and two air temperatures (current, (current + 4°C)). Gravimetric water use, leaf gas exchange and tissue dry mass and %N were determined. Solid-state 15N-NMR spectroscopy was used for determining the partitioning of N chemical groups in the dry matter fractions. Water use efficiency (WUE) improved with increasing [CO2] at ambient temperature, but strong leaf area and weak reductions in transpiration rates led to greater water use at elevated [CO2]. High temperature increased plant water use, such that WUE was not significantly stimulated by increasing [CO2] at high temperature. Total N uptake increased with increasing [CO2] but not temperature, less than the increase recorded for plant biomass. Tissue [N] decreased with rising [CO2] and at high temperature, but N use efficiency increased with rising [CO2]. Total N uptake was positively correlated with total water use and root biomass under all treatments. Growth [CO2] and temperature did not affect the partitioning of 15N among the N chemical groups. The reductions of tissue [N] with [CO2] and temperature were generic, not specific to particular N compounds. The results suggest that reductions in tissue [N] are caused by changes in root N uptake by mass flow due to altered transpiration rates at elevated [CO2] and temperature.