Genotypic variability was studied in two Mediterranean grass species, Bromus erectus and Dactylis glomerata, with regard to the response to CO2 of leaf total non-structural carbohydrate concentration ([TNC]lf), specific leaf area (SLA), and leaf carbon and nitrogen concentrations ([C]lf and [N]lf, respectively). Fourteen genotypes of each species were grown together on intact soil monoliths at ambient and elevated CO2 concentrations (350 and 700 μmol mol−1, respectively). In both species, the most consistent effect of elevated CO2 was an increase in [TNC]lf and a decrease in leaf nitrogen concentration when expressed either as total dry mass [Nm]lf, structural dry mass [Nmst]lf or leaf area [Na]lf. The SLA decreased only in D. glomerata, due to an accumulation of total non-structural carbohydrates and to an increase in leaf density. No genotypic variability was found for any variable in B. erectus, suggesting that genotypes responded in a similar way to elevated CO2. In D. glomerata, a genotypic variability was found only for [Cst], [Nm]lf, [Nmst]lf and [Na]lf. Since [Nm]lf is related to plant growth and is a strong determinant of plant–herbivore interactions, our results suggest evolutionary consequences of elevated CO2 through competitive interactions or herbivory.