The evolution of diffusive and biochemical capacities for photosynthesis was predominantly shaped by [CO₂] with a smaller contribution from [O₂]

The atmospheric concentration of carbon dioxide ([CO₂]) and oxygen ([O₂]) directly influence rates of photosynthesis (P_N) and photorespiration (R_PR) through the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO). Levels of [CO₂] and [O₂] have varied over Earth history affecting rates of both CO₂ uptake and loss, alongside associated transpirative water-loss. The availability of CO₂ has likely acted as a stronger selective pressure than [O₂] due to the greater specificity of RubisCO for CO₂. The role of [O₂], and the interaction of [O₂] and [CO₂], in plant evolutionary history is less understood. We exposed twelve phylogenetically diverse species to combinations of sub-ambient, ambient and super-ambient [O₂] and [CO₂] to examine the biochemical and diffusive components of P_N and the possible role of [O₂] as a selective pressure. Photosynthesis, photosynthetic capacity and stomatal, mesophyll and total conductance to CO₂ were higher in the derived eudicot and monocot angiosperms than the more basal ferns, gymnosperms and basal angiosperms which originated in atmospheres characterised by higher CO₂:O₂ ratios. The ratio of R_PR:P_N was lower in the monocots, consistent with greater carboxylation capacity and higher stomatal and mesophyll conductance making easier CO₂ delivery to chloroplasts. The effect of [O₂] and [CO₂] on P_N/R_PR was less evident in more derived species with a higher conductance to CO₂. The effect of [O₂] was less apparent at high [CO₂], suggesting that atmospheric [O₂] may only have exerted a strong selective pressure on plant photosynthetic processes during periods characterised by low atmospheric CO₂:O₂ ratios. Current rising [CO₂] will predominantly enhance P_N rates in species with low diffusive conductance to CO₂.