Nitrogen content per unit leaf area (Narea) is a key variable in plant functional ecology and biogeochemistry. Narea comprises a structural component, which scales with leaf mass per area (LMA), and a metabolic component, which scales with Rubisco capacity. The co-ordination hypothesis, as implemented in LPJ and related global vegetation models, predicts that Rubisco capacity should be directly proportional to irradiance but should decrease with increases in ci : ca and temperature because the amount of Rubisco required to achieve a given assimilation rate declines with increases in both. We tested these predictions using LMA, leaf δ13C, and leaf N measurements on complete species assemblages sampled at sites on a north–south transect from tropical to temperate Australia. Partial effects of mean canopy irradiance, mean annual temperature, and ci : ca (from δ13C) on Narea were all significant and their directions and magnitudes were in line with predictions. Over 80 % of the variance in community-mean (ln) Narea was accounted for by these predictors plus LMA. Moreover, Narea could be decomposed into two components, one proportional to LMA (slightly steeper in N-fixers), and the other to Rubisco capacity as predicted by the co-ordination hypothesis. Trait gradient analysis revealed ci : ca to be perfectly plastic, while species turnover contributed about half the variation in LMA and Narea.