Testing unified theories for ozone response in C₄ species

There is tremendous interspecific variability in O₃  sensitivity among C₃  species, but variation among C₄  species has been less clearly documented. It is also unclear whether stomatal conductance and leaf structure such as leaf mass per area (LMA) determine the variation in sensitivity to O₃ across species. In this study, we investigated leaf morphological, chemical, and photosynthetic responses of 22 genotypes of four C₄ bioenergy species (switchgrass, sorghum, maize, and miscanthus) to elevated O₃ in side-by-side field experiments using free-air O₃ concentration enrichment (FACE). The C₄  species varied largely in leaf morphology, physiology, and nutrient composition. Elevated O₃ did not alter leaf morphology, nutrient content, stomatal conductance, chlorophyll fluorescence, and respiration in most genotypes but reduced net CO₂ assimilation in maize and photosynthetic capacity in sorghum and maize. Species with lower LMA and higher stomatal conductance tended to show greater losses in photosynthetic rate and capacity in elevated O₃ compared with species with higher LMA and lower stomatal conductance. Stomatal conductance was the strongest determinant of leaf photosynthetic rate and capacity. The response of both area- and mass-based leaf photosynthetic rate and capacity to elevated O₃ were not affected by LMA directly but negatively influenced by LMA indirectly through stomatal conductance. These results demonstrate that there is significant variation in O₃  sensitivity among C₄  species with maize and sorghum showing greater sensitivity of photosynthesis to O₃ than switchgrass and miscanthus. Interspecific variation in O₃  sensitivity was determined by direct effects of stomatal conductance and indirect effects of LMA. This is the first study to provide a test of unifying theories explaining variation in O₃  sensitivity in C₄ bioenergy grasses. These findings advance understanding of O₃ tolerance in C₄  grasses and could aid in optimal placement of diverse C₄ bioenergy feedstock across a polluted landscape.