Models of stomatal conductance (gs) are based on coupling between gs and CO2 assimilation (Anet), and it is often assumed that the slope of this relationship ('g1') is constant across species. However, if different plant species have adapted to different access costs of water, then there will be differences in g1 among species. We hypothesized that g1 should vary among species adapted to different climates, and tested the theory and its linkage to plant hydraulics using four Eucalyptus species from different climatic origins in a common garden. Optimal stomatal theory predicts that species from sub-humid zones have a lower marginal water cost of C gain, hence lower g1 than humid-zone species. In agreement with the theory that g1 is related to tissue carbon costs for water supply, we found a relationship between wood density and g1 across Eucalyptus species of contrasting climatic origins. There were significant reductions in the parameter g1 during drought in humid but not sub-humid species, with the latter group maintaining g1 in drought. There are strong differences in stomatal behaviour among related tree species in agreement with optimal stomatal theory, and these differences are consistent with the economics involved in water uptake and transport for carbon gain.