Background and aims The survival and coexistence of plants in water-limited environments are related to their ability to coordinate water acquisition and regulation of water loss. To assess the coordination among below and aboveground hydraulic traits and the diversity of water-use strategies, we evaluated rooting depth and several leaf hydraulic traits of 15 species in campos rupestres, a seasonally-dry biodiversity hotspot in central Brazil. Methods We assessed the depth of plant water acquisition by excavating roots and analyzing the stable isotope composition of hydrogen (δD) and oxygen (δ18O) in the xylem and soil water. We also measured mid-morning stomatal conductance, leaf-water potential at turgor loss point (ΨTLP) and pre-dawn leaf water potentials (ΨPD) during wet and dry seasons. Results We demonstrated that rooting depth is a good predictor of seasonal variations in stomatal conductance and ΨPD. Shallow-rooted plants had greater variation in stomatal conductance and ΨPD than deep-rooted plants. Woody plants with shallower roots also had lower ΨTLP than deep-rooted plants, revealing higher drought resistance. Conclusion We demonstrate that shallow-rooted species, more exposed to variation in water availability, have mechanisms to confer drought resistance through turgor maintenance. Our results support the theory of hydrological niche segregation and its underlying trade-offs related to drought resistance.