Optimal coordination and reorganization of photosynthetic properties in C₄ grasses

Each of >20 independent evolutions of C₄ photosynthesis in grasses required reorganization of the Calvin-Benson-cycle (CB-cycle) within the leaf, along with coordination of C₄ -cycle enzymes with the CB-cycle to maximize CO₂ assimilation. Considering the vast amount of time over which C₄ evolved, we hypothesized (i) trait divergences exist within and across lineages with both C₄ and closely related C₃ grasses, (ii) trends in traits after C₄ evolution yield the optimization of C₄ through time, and (iii) the presence/absence of trends in coordination between the CB-cycle and C₄ -cycle provides information on the strength of selection. To address these hypotheses, we used a combination of optimality modelling, physiological measurements and phylogenetic-comparative-analysis. Photosynthesis was optimized after the evolution of C₄ causing diversification in maximal assimilation, electron transport, Rubisco carboxylation, phosphoenolpyruvate carboxylase and chlorophyll within C₄ lineages. Both theory and measurements indicated a higher light-reaction to CB-cycle ratio (J_atpmax /V_cmax ) in C₄ than C₃ . There were no evolutionary trends with photosynthetic coordination between the CB-cycle, light reactions and the C₄ -cycle, suggesting strong initial selection for coordination. The coordination of CB-C₄ -cycles (V_pmax /V_cmax ) was optimal for CO₂ of 200 ppm, not to current conditions. Our model indicated that a higher than optimal V_pmax /V_cmax affects assimilation minimally, thus lessening recent selection to decrease V_pmax /V_cmax .