Ozone (O3) is a phytotoxic air pollutant, the adverse effects of which on growth and photosynthesis are modified by other environmental factors. In this study, we examined the combined effects of O3, elevated CO2, and soil nitrogen supply on Siebold's beech seedlings. Seedlings were grown under combinations of two levels of O3 (low and two times ambient O3 concentration), two levels of CO2 (ambient and 700 ppm), and three levels of soil nitrogen supply (0, 50, and 100 kg N ha−1 year−1) during two growing seasons (2019 and 2020), with leaf photosynthetic traits being determined during the second season. We found that elevated CO2 ameliorated O3-induced reductions in photosynthetic activity, whereas the negative effects of O3 on photosynthetic traits were enhanced by soil nitrogen supply. We observed three-factor interactions in photosynthetic traits, with the ameliorative effects of elevated CO2 on O3-induced reductions in the maximum rate of carboxylation being more pronounced under high than under low soil nitrogen conditions in July. In contrast, elevated CO2-induced amelioration of the effects of O3 on stomatal function-related traits was more pronounced under low soil nitrogen conditions. Although we observed several two- or three-factor interactions of gas and soil treatments with respect to leaf photosynthetic traits, the shoot to root dry mass (S/R) ratio was the only parameter for which a significant interaction was detected among seedling growth parameters. O3 caused a significant increase in S/R under ambient CO2 conditions, whereas no similar effects were observed under elevated CO2 conditions. Collectively, our findings reveal the complex interactive effects of elevated CO2 and soil nitrogen supply on the detrimental effects of O3 on leaf photosynthetic traits, and highlight the importance of taking into consideration differences between the responses of CO2 uptake and growth to these three environmental factors.
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