Reduced ascorbate pool and its maintenance are important determinants of O3 damage to net photosynthetic rate in Fagus crenata under elevated CO2 and soil N supply

Many studies have reported modification in the degree of O3 damage to photosynthesis by elevated CO2 and soil N supply. However, the mechanism underlying the modification is unclear. To clarify the important determinants in the degree of O3 damage to net photosynthetic rate (A) in the leaves of Fagus crenata (Siebold's beech) under elevated CO2 and with different soil N supply, F. crenata seedlings were grown for two growing seasons under combinations of two O3 levels (low concentration at approximately 4 nmol mol-1 and two times the ambient concentration), two CO2 levels (ambient and 700 mu mol mol-1), and three levels of soil N supply (0, 50 and 100 kg N ha- 1 year -1). During the second growing season, we determined A, stomatal conductance for calculating phytotoxic O3 dose (POD), antioxidant concentrations, and antioxidative enzyme activities in the leaves for evaluating O3 detoxification capacity. We calculated the O3-induced reduction in mean A (Delta Amean) during the second growing season using the data reported in our previous study and plotted it against mean daily POD without flux threshold (POD0). There was no significant linear nor non-linear relationship, suggesting that not only POD0 but also O3 detoxification capacity are important determinants of Delta Amean under elevated CO2 and N supply. We found significant negative linear relationships of Delta Amean per unit POD0 (Delta Amean/POD0) with reduced ascorbate concentration in the low O3 treatment, and with percentage of O3-induced change in activity of monodehydroascorbate reductase (MDAR). In addition, the Delta Amean/POD0 was positively and significantly correlated with the activity ratio of ascorbate peroxidase to MDAR. These results suggest that reduced ascorbate pool and its maintenance through the action of MDAR could be important determinants in the degree of O3 damage to net photosynthesis under elevated CO2 and soil N supply.