Karst and non-karst forests occur in the same region in south-west China, but the soil water and mineral nutrients availability are different between the forests. Our hypothesis was that the leaves of karst trees would be better adapted to dry, nutrient-poor conditions than those of trees in a nearby non-karst forest. We compared the gas exchange, anatomical characteristics and mineral nutrient concentrations in leaves from 21 tree species in a tropical karst forest and 19 species in a nearby non-karst forest in south-west China. We found that the leaves of karst trees had higher P concentrations, photosynthetic capacity and water use efficiency, and greater adaxial and abaxial epidermis thickness than leaves of non-karst forest trees. Evergreen and deciduous trees differed more significantly in leaf functional traits in the karst forest than in the non-karst forest. The leaf palisade: spongy mesophyll thickness ratio was positively correlated with stomatal conductance and negatively correlated with photosynthetic water use efficiency in the karst forest but not in the non-karst forest. Our findings indicate that karst forest trees are more conservative in water use, whereas soil P deficiency could be a major limiting factor for the growth of non-karst forest trees.