Experiments were conducted to evaluate root traits associated with nutrient exploitation following defoliation in three coexisting perennial grasses in a semi-arid savanna. Root length density was determined within soil cores directly beneath plants, nitrogen uptake was evaluated by excised-root assay with (15NH4)2SO4, and mycorrhizal root colonization was estimated by observation of root segments. Root length density was lowest for Bouteloua curtipendula, intermediate for Eriochloa sericea, and highest for Aristida purpurea indicating that root length density was a more important trait for the mid-seral than the late-seral species. Rates of 15N uptake were greatest in the least grazing tolerant late-seral species, E. sericea, intermediate in the mid-seral species, A. purpurea, and lowest in the most grazing tolerant late-seral species, B. curtipendula. Two successive defoliations reduced 15N uptake 60% in the late-seral species with the greatest uptake rate (E. sericea), but not in species with lowest uptake rates (B. curtipendula). Root length colonization was consistently high (33–61%) in all three species suggesting that these C4 perennial grasses may function as obligate mycotrophs. Contrasting responses among the two late-seral species indicate that the least grazing tolerant species, E. sericea, appears best adapted for nutrient exploitation while the most grazing tolerant species, B. curtipendula, appears best adapted for efficient nutrient retention. Contrasting responses of nitrogen uptake to short-term defoliation parallel the population responses of these two coexisting late-seral species to long-term herbivory. These data indicate that herbivory may shift interspecific competitive interactions by mediating nutrient exploitation and that a trade-off may exist between nutrient exploitation and herbivory tolerance in these species.