Leaf anatomy does not explain the large variability of mesophyll conductance across C₃ crop species

Increasing mesophyll conductance of CO₂ (g_m ) is a strategy to improve photosynthesis in C₃ crops. However, the relative importance of different anatomical traits in determining g_m in crops is unclear. Mesophyll conductance measurements were performed on 10 crops using the online carbon isotope discrimination method and the 'variable J' method in parallel. The influences of crucial leaf anatomical traits on g_m were evaluated using a one-dimensional anatomical CO₂ diffusion model. The g_m values measured using two independent methods were compatible, although significant differences were observed in their absolute values. Quantitative analysis showed that cell wall thickness and chloroplast stroma thickness are the most important elements along the diffusion pathway. Unexpectedly, the large variability of g_m across crops was not associated with any investigated leaf anatomical traits except chloroplast thickness. The g_m values estimated using the anatomical model differed remarkably from the values measured in vivo in most species. However, when the species-specific effective porosity of the cell wall and the species-specific facilitation effect of CO₂ diffusion across the membrane and chloroplast stoma were taken into account, the model could output g_m values very similar to those measured in vivo. These results indicate that g_m variation across crops is probably also driven by the effective porosity of the cell wall and effects of facilitation of CO₂ transport across the membrane and chloroplast stroma in addition to the thicknesses of the elements.