Trees adapted to mesic and xeric habits may differ in a suite of physiological responses that affect leaf-level carbon balance, including the relationship between photosynthesis (A) and respiration at night (R(n)). Understanding the factors that regulate physiological function in mesic and xeric species is critical for predicting changes in growth and distribution under changing climates. In this study, we examined the relationship between A and R(n), and leaf traits that may regulate A and R(n), in six Eucalyptus species native to mesic or xeric ecosystems, during two 24-h cycles in a common garden under high soil moisture. Peak A and R(n) generally were higher in xeric compared with mesic species. Across species, A and R(n) covaried, correlated with leaf mass per area, leaf N per unit area and daytime soluble sugar accumulation. A also covaried with g(s), which accounted for 93% of the variation in A within species. These results suggest that A and R(n) in these six Eucalyptus species were linked through leaf N and carbohydrates. Further, the relationship between A and R(n) across species suggests that differences in this relationship between mesic and xeric Eucalyptus species in their native habitats may be largely driven by environmental factors rather than inter-specific genetic variation.