The loss of foliage through pruning of live branches may reduce tree growth or it may be compensated by photosynthetic up-regulation of the remaining crown. Here, the changes in light-saturated photosynthesis following pruning to remove 50% of green crown length were examined in 4-year-old Eucalyptus pilularis Sm. and Eucalyptus cloeziana F. Muell. trees. The objectives of the study were to: (1) compare leaf-level physiological (light-saturated photosynthesis ( A max), stomatal conductance ( g), transpiration ( T), dark respiration ( R d), quantum yield ( Φ), light compensation point ( Γ), water-use efficiency (WUE), nitrogen-use efficiency (NUE)) traits in species with contrasting crown dynamics and structure, (2) examine the effect of crown position on these traits, and (3) examine the effect of pruning on A max, g, T, WUE, NUE, leaf N and P concentrations and specific leaf area (SLA). Prior to pruning there were no differences in R d, Γ and Φ between E. pilularis and E. cloeziana but differences in A max, T, g, leaf N, leaf P, WUE, NUE and SLA. Whereas the rate of physiological processes ( A max, T, and g) and leaf N and P concentrations increased with crown height, R d, Γ, Φ and SLA declined along this vertical gradient, except in the upper crown of E. cloeziana where A max, T and g were not different to the lower crown. No up-regulation of photosynthesis or changes in leaf physiology occurred between 6 and 13 months after pruning in either species. The results provide an important basis for modelling pruning effects in process-based tree growth models.