Large spatial and temporal gradients in rainfall and temperature occur across Australia. This heterogeneity drives ecological differentiation in vegetation structure and ecophysiology. We examined multiple leaf-scale traits, including foliar C isotope discrimination (Δ C), rates of photosynthesis and foliar N concentration and their relationships with multiple climate variables. Fifty-five species across 27 families were examined across eight sites spanning contrasting biomes. Key questions addressed include: (i) Does Δ C and intrinsic water-use efficiency (WUE ) vary with climate at a continental scale? (ii) What are the seasonal and spatial patterns in Δ C/WUE across biomes and species? (iii) To what extent does Δ C reflect variation in leaf structural, functional and nutrient traits across climate gradients? and (iv) Does the relative importance of assimilation and stomatal conductance in driving variation in Δ C differ across seasons? We found that MAP, temperature seasonality, isothermality and annual temperature range exerted independent effects on foliar Δ C/WUE . Temperature-related variables exerted larger effects than rainfall-related variables. The relative importance of photosynthesis and stomatal conductance (g ) in determining Δ C differed across seasons: Δ C was more strongly regulated by g during the dry-season and by photosynthetic capacity during the wet-season. Δ C was most strongly correlated, inversely, with leaf mass area ratio among all leaf attributes considered. Leaf N was significantly and positively correlated with MAP during dry- and wet-seasons and with moisture index (MI) during the wet-season but was not correlated with Δ C. Leaf P showed significant positive relationship with MAP and Δ C only during the dry-season. For all leaf nutrient-related traits, the relationships obtained for Δ C with MAP or MI indicated that Δ C at the species level reliably reflects the water status at the site level. Temperature and water availability, not foliar nutrient content, are the principal factors influencing Δ C across Australia.

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