Stomatal and non-stomatal limitations in savanna trees and C₄ grasses grown at low, ambient and high atmospheric CO₂

By the end of the century, atmospheric CO₂ concentration ([CO₂]_a) could reach 800 ppm, having risen from ∼200 ppm ∼24 Myr ago. Carbon dioxide enters plant leaves through stomata that limit CO₂ diffusion and assimilation, imposing stomatal limitation (L_S). Other factors limiting assimilation are collectively called non-stomatal limitations (L_NS). C₄ photosynthesis concentrates CO₂ around Rubisco, typically reducing L_S. C₄-dominated savanna grasslands expanded under low [CO₂]_a and are metastable ecosystems where the response of trees and C₄ grasses to rising [CO₂]_a will determine shifting vegetation patterns. How L_S and L_NS differ between savanna trees and C₄ grasses under different [CO₂]_a will govern the responses of CO₂ fixation and plant cover to [CO₂]_a – but quantitative comparisons are lacking. We measured assimilation, within soil wetting–drying cycles, of three C₃ trees and three C₄ grasses grown at 200, 400 or 800 ppm [CO₂]_a. Using assimilation–response curves, we resolved L_S and L_NS and show that rising [CO₂]_a alleviated L_S, particularly for the C₃ trees, but L_NS was unaffected and remained substantially higher for the grasses across all [CO₂]_a treatments. Because L_NS incurs higher metabolic costs and recovery compared with L_S, our findings indicate that C₄ grasses will be comparatively disadvantaged as [CO₂]_a rises.