Ozone exposure alters nutrients and stoichiometric ratios in different organs of four urban tree species despite limited negative effects on leaf physiology and plant growth and biomass

Abstract

To better understand the effects of ground-level ozone (O₃) on nutrients and stoichiometry in different plant organs, urban tree species Celtis sinensis, Cyclocarya paliurus, Quercus acutissima, and Quercus nuttallii were subjected to a constant exposure to charcoal-filtered air (CF), nonfiltered air (NF), or NF + 40, 60, or 80 nmol O₃ mol^–1 (NF40, NF60, and NF80) starting early in the summer of the growing season. At the end of summer, net CO₂ assimilation rate (A), stomatal conductance (g_s), leaf mass per area (LMA), and/or leaf greenness (SPAD) either were not significantly affected by elevated O₃ or were even higher in some cases during the summer compared with the CF or NF controls. LMA was significantly lower in autumn only after the highest O₃ exposures. Compared to NF, NF40 caused a large increase in g_s across species in late summer and more K and Mn in stems. At the end of the growing season, nutrient status and stoichiometric ratios in different organs were variously altered under O₃ stress; many changes were large and often species-specific. Across O₃ treatments, LMA was primarily associated with C and Mg levels in leaves and Ca levels in leaves and stems. NF40 enriched K, P, Fe, and Mn in stems, relative to NF, and NF60 enhanced Ca in leaves relative to CF and NF40. Moreover, NF resulted in a higher Ca/Mg ratio in leaves of Q. acutissima only, relative to the other O₃ regimes. Interestingly, across species, O₃ stress led to different nutrient modifications in different organs (stems + branches vs leaves). Thus, ambient and/or elevated O₃ exposures can alter the dynamics and distribution of nutrients and disrupt stoichiometry in different organs in a species-specific manner. Changes in stoichiometry reflect an important defense mechanism in plants under O₃, and O₃ pollution adds more risk to ecological stoichiometries in urban areas.

Keywords

Air pollution / Ground-level ozone / Urban green / Micronutrients / Nutrient cycling

References

[1] Agathokleous E, Saitanis CJ. Plant susceptibility to ozone: a tower of babel?. Sci Total Environ, 2020, 703 134962

[2] Agathokleous E, Saitanis CJ, Stamatelopoulos D, Mouzaki-Paxinou AC, Paoletti E, Manning WJ. Olive oil for dressing plant leaves so as to avoid O₃ injury. Water Air Soil Pollut, 2016, 227(8): 282

[3] Agathokleous E, Kitao M, Chu QN, Saitanis CJ, Paoletti E, Manning WJ, Watanabe T, Koike T. Effects of ozone (O₃) and ethylenediurea (EDU) on the ecological stoichiometry of a willow grown in a free-air exposure system. Environ Pollut, 2018, 238: 663-676

[4] Agathokleous E, Kitao M, Koike T. Ethylenediurea (EDU) effects on hybrid larch saplings exposed to ambient or elevated ozone over three growing seasons. J for Res, 2022, 33(1117-135

[5] Agathokleous E, Saitanis CJ, Agathokleous S, Savvides C, Calatayud V, Sicard P. isoAOT40: an improved ozone exposure index based on the Annual Ozone Spectrum Profile (AO3SP). J for Res, 2022, 33(6): 1949-1955

[6] Alejandro S, Höller S, Meier B, Peiter E. Manganese in plants: from acquisition to subcellular allocation. Front Plant Sci, 2020, 11: 300

[7] Allen AP, Gillooly JF. Towards an integration of ecological stoichiometry and the metabolic theory of ecology to better understand nutrient cycling. Ecol Lett, 2009, 12(5): 369-384

[8] Badia A, Vidal V, Ventura S, Curcoll R, Segura R, Villalba G. Modelling the impacts of emission changes on O₃ sensitivity, atmospheric oxidation capacity, and pollution transport over the Catalonia region. Atmos Chem Phys, 2023, 23(1810751-10774

[9] Bhattarai H, Tai APK, Val Martin M, Yung DHY. Impacts of changes in climate, land use, and emissions on global ozone air quality by mid-21st century following selected Shared Socioeconomic Pathways. Sci Total Environ, 2024, 906 167759

[10] Bloom AJ, Lancaster KM. Manganese binding to Rubisco could drive a photorespiratory pathway that increases the energy efficiency of photosynthesis. Nat Plants, 2018, 4(7): 414-422

[11] Box GEP, Cox DR. An analysis of transformations. J R Stat Soc Ser B Stat Methodol, 1964, 26(2): 211-243

[12] Calabrese EJ, Mattson MP. How does hormesis impact biology, toxicology, and medicine?. NPJ Aging Mech Dis, 2017, 3: 13

[13] Cao JX, Shang H, Chen Z, Tian Y, Yu H. Effects of elevated ozone on stoichiometry and nutrient pools of Phoebe bournei (hemsl.) Yang and Phoebe zhennan S lee et F. N. Wei seedlings in subtropical China. Forests, 2016, 7(4): 78

[14] Carvalho MEA, Castro PRC, Azevedo RA. Hormesis in plants under Cd exposure: from toxic to beneficial element?. J Hazard Mater, 2020, 384 121434

[15] Chen ZL, Jian YQ, Wu Q, Wu J, Sheng WB, Jiang S, Shehla N, Aman S, Wang W. Cyclocarya paliurus (Batalin) Iljinskaja: botany, ethnopharmacology, phytochemistry and pharmacology. J Ethnopharmacol, 2022, 285 114912

[16] Cohen J. Statistical power analysis for the behavioral sciences, 2013Routledge

[17] Costantini D. Hormesis promotes evolutionary change. Dose Response, 2019, 17(21559325819843376

[18] Costantini D. Costantini D, Marasco V. For better or worse: benefits and costs of transgenerational plasticity and the transhormesis hypothesis. Development Strategies and Biodiversity: Darwinian Fitness and Evolution in the Anthropocene, 2022, Cham, Springer International Publishing37-49

[19] Costantini D, Borremans B. The linear no-threshold model is less realistic than threshold or hormesis-based models: an evolutionary perspective. Chem Biol Interact, 2019, 301: 26-33

[20] Costantini D, Metcalfe NB, Monaghan P. Ecological processes in a hormetic framework. Ecol Lett, 2010, 13(111435-1447

[21] Cotrozzi L. The effects of tropospheric ozone on oaks: a global meta-analysis. Sci Total Environ, 2021, 756 143795

[22] Elansary HO, Szopa A, Kubica P, Ekiert H, Mattar MA, Al-Yafrasi MA, El-Ansary DO, El-Abedin TKZ, Yessoufou K. Polyphenol profile and pharmaceutical potential of Quercus spp. bark extracts. Plants, 2019, 8(11): 486

[23] Elser JJ, Fagan WF, Kerkhoff AJ, Swenson NG, Enquist BJ. Biological stoichiometry of plant production: metabolism, scaling and ecological response to global change. New Phytol, 2010, 186(3593-608

[24] Emberson L. Effects of ozone on agriculture, forests and grasslands. Philos Trans A Math Phys Eng Sci, 2020, 378(2183): 20190327

[25] Erofeeva EA. Plant hormesis and Shelford’s tolerance law curve. J Forestry Res, 2021, 32(51789-1802

[26] Erofeeva EA. Environmental hormesis: from cell to ecosystem. Curr Opin Environ Sci Health, 2022, 29 100378

[27] Erofeeva EA, Gelashvili DB, Rozenberg GS. The modern concept of hormesis: an overview of the issue and its significance for ecology. Biol Bull Rev, 2023, 13(3): S229-S239

[28] Esmeijer-Liu AJ, Aerts R, Kürschner WM, Bobbink R, Lotter AF, Verhoeven JTA. Nitrogen enrichment lowers Betula pendula green and yellow leaf stoichiometry irrespective of effects of elevated carbon dioxide. Plant Soil, 2009, 316(1): 311-322

[29] Fu W, He XY, Xu S, Chen W, Li Y, Li B, Su LL, Ping Q. Changes in nutrients and decay rate of Ginkgo biloba leaf litter exposed to elevated O₃ concentration in urban area. PeerJ, 2018, 6 e4453

[30] Gao YQ, Yuan Y, Li QK, Kou L, Fu XL, Dai XQ, Wang HM. Mycorrhizal type governs foliar and root multi-elemental stoichiometries of trees mainly via root traits. Plant Soil, 2021, 460(1): 229-246

[31] Geihs MA, Moreira DC, López-Martínez G, Minari M, Ferreira-Cravo M, Carvajalino-Fernández JM, Hermes-Lima M. Commentary: ultraviolet radiation triggers “preparation for oxidative stress” antioxidant response in animals: similarities and interplay with other stressors. Comp Biochem Physiol Part A Mol Integr Physiol, 2020, 239 110585

[32] Gupta A, Yadav DS, Agrawal SB, Agrawal M. Sensitivity of agricultural crops to tropospheric ozone: a review of Indian researches. Environ Monit Assess, 2022, 194(12): 894

[33] Gupta A, Yadav DS, Agrawal SB, Agrawal M. Individual effects of high temperature and tropospheric ozone on tomato: a review. J Plant Growth Regul, 2023, 42(31421-1443

[34] Hopkins WG (2016) A New View of Statistics [WWW Document]. URL http://newstats.org (accessed 10.4.24)

[35] Hoshika Y, Agathokleous E, Moura BB, Paoletti E. Ozone risk assessment with free-air controlled exposure (FACE) experiments: a critical revisit. Environ Res, 2024, 255 119215

[36] Hoshika Y, Moura BB, Cotrozzi L, Nali C, Alfarraj S, Rennenberg H, Paoletti E. An assessment of ozone risk for date palm suggests that phytotoxic ozone dose nonlinearly affects carbon gain. Environ Pollut, 2024, 342 123143

[37] Imtiaz H, Mir AR, Corpas FJ, Hayat S. Impact of potassium starvation on the uptake, transportation, photosynthesis, and abiotic stress tolerance. Plant Growth Regul, 2023, 99(3): 429-448

[38] Ishfaq M, Wang YQ, Yan MW, Wang Z, Wu LQ, Li CJ, Li XX. Physiological essence of magnesium in plants and its widespread deficiency in the farming system of China. Front Plant Sci, 2022, 13 802274

[39] Ivatt PD, Evans MJ, Lewis AC. Suppression of surface ozone by an aerosol-inhibited photochemical ozone regime. Nat Geosci, 2022, 15: 536-540

[40] Jing T, Li JY, He YD, Shankar A, Saxena A, Tiwari A, Maturi KC, Solanki MK, Singh V, Eissa MA, Ding ZL, Xie JH, Awasthi MK. Role of calcium nutrition in plant physiology: advances in research and insights into acidic soil conditions—a comprehensive review. Plant Physiol Biochem, 2024, 210 108602

[41] Karabourniotis G, Liakopoulos G, Nikolopoulos D, Bresta P, Stavroulaki V, Sumbele S. “Carbon gain vs. water saving, growth vs. defence”: two dilemmas with soluble phenolics as a joker. Plant Sci, 2014, 227: 21-27

[42] Khan F, Siddique AB, Shabala S, Zhou MX, Zhao CC. Phosphorus plays key roles in regulating plants' physiological responses to abiotic stresses. Plants, 2023, 12(15): 2861

[43] Koerselman W, Meuleman AFM. The vegetation N: P ratio: a new tool to detect the nature of nutrient limitation. J Appl Ecol, 1996, 33(61441-1450

[44] Krohling CA, Eutrópio FJ, Bertolazi AA, Dobbss LB, Campostrini E, Dias T, Ramos AC. Ecophysiology of iron homeostasis in plants. Soil Sci Plant Nutr, 2016, 62(1): 39-47

[45] Lee Jones A, Ormondroyd A, Hayes F, Jeffers ES. Reflections of stress: Ozone damage in broadleaf saplings can be identified from hyperspectral leaf reflectance. Environ Pollut, 2024, 360 124642

[46] Li L, Wang XK, Niu JF, Cui J, Zhang QQ, Wan WX, Liu BJ. Effects of elevated atmospheric O₃ concentrations on early and late leaf growth and elemental contents of Acer truncatum Bung under mild drought. Acta Ecol Sin, 2017, 37(1): 31-34

[47] Li P, Feng ZZ, Catalayud V, Yuan XY, Xu YS, Paoletti E. A meta-analysis on growth, physiological, and biochemical responses of woody species to ground-level ozone highlights the role of plant functional types. Plant Cell Environ, 2017, 40(10): 2369-2380

[48] Li S, Yang Y, Wang HL, Li PW, Li K, Ren LL, Wang PY, Li BJ, Mao YH, Liao H. Rapid increase in tropospheric ozone over Southeast Asia attributed to changes in precursor emission source regions and sectors. Atmos Environ, 2023, 304 119776

[49] Lim YJ, Kwak MJ, Lee J, Kang D, Je SM, Woo SY. Korean flowering cherry (Prunus × yedoensisMatsum.) response to elevated ozone: physiological traits and biogenic volatile organic compounds emission. Hortic Environ Biotechnol, 2024, 65(6): 1025-1042

[50] Lyu XP, Li K, Guo H, Morawska L, Zhou BN, Zeren YZ, Jiang F, Chen CH, Goldstein AH, Xu XB, Wang T, Lu X, Zhu T, Querol X, Chatani S, Latif MT, Schuch D, Sinha V, Kumar P, Mullins B, Seguel R, Shao M, Xue LK, Wang N, Chen JM, Gao J, Chai FH, Simpson I, Sinha B, Blake DR. A synergistic ozone-climate control to address emerging ozone pollution challenges. One Earth, 2023, 6(8): 964-977

[51] Manzini J, Hoshika Y, Danti R, Moura BB, Paoletti E, Della Rocca G. Ozone risk assessment of common cypress (Cupressus sempervirens L.) clones and effects of Seiridium cardinale infection. J Environ Sci, 2024, 151: 441-453

[52] Matzek V, Vitousek PM. N: P stoichiometry and protein: RNA ratios in vascular plants: an evaluation of the growth-rate hypothesis. Ecol Lett, 2009, 12(8): 765-771

[53] Nakaji T, Kobayashi T, Kuroha M, Omori K, Matsumoto Y, Yonekura T, Watanabe K, Utriainen J, Izuta T. Growth and nitrogen availability of red pine seedlings under high nitrogen load and elevated ozone. Water Air Soil Pollut Focus, 2004, 4(2): 277-287

[54] Paoletti E, Sicard P, Hoshika Y, Fares S, Badea O, Pitar D, Popa I, Anav A, Moura BB, De Marco A. Towards long-term sustainability of stomatal ozone flux monitoring at forest sites. Sustain Horiz, 2022, 2 100018

[55] Paoletti E, Pagano M, Zhang L, Badea O, Hoshika Y. Allocation of nutrients and leaf turnover rate in poplar under ambient and enriched ozone exposure and soil nutrient manipulation. Biology, 2024, 13(4): 232

[56] Pérez-Harguindeguy N, Díaz S, Vendramini F, Cornelissen JHC, Gurvich DE, Cabido M. Leaf traits and herbivore selection in the field and in Cafeteria experiments. Austral Ecol, 2003, 28(6642-650

[57] Persson J, Fink P, Goto A, Hood JM, Jonas J, Kato S. To be or not to be what you eat: regulation of stoichiometric homeostasis among autotrophs and heterotrophs. Oikos, 2010, 119: 741-751

[58] Qi ZP, Fang XY, Xie YY, Wang L, Zhang YW, Zhao LG. Bioassay-guided isolation of anti-inflammatory constituents from Celtissinensis leaves. J Food Biochem, 2021, 45(1 e13580

[59] Querol X, Massagué J, Alastuey A, Moreno T, Gangoiti G, Mantilla E, Duéguez JJ, Escudero M, Monfort E, Pérez García-Pando C, Petetin H, Jorba O, Vázquez V, de la Rosa J, Campos A, Muñóz M, Monge S, Hervás M, Javato R, Cornide MJ. Lessons from the COVID-19 air pollution decrease in Spain: Now what?. Sci Total Environ, 2021, 779 146380

[60] Schmidt SB, Husted S. The biochemical properties of manganese in plants. Plants, 2019, 8(10): 381

[61] Sebastiano M, Messina S, Marasco V, Costantini D. Hormesis in ecotoxicological studies: a critical evolutionary perspective. Curr Opin Toxicol, 2022, 29: 25-30

[62] Shang B, Feng ZZ, Li P, Calatayud V. Elevated ozone affects C, N and P ecological stoichiometry and nutrient resorption of two poplar clones. Environ Pollut, 2018, 234: 136-144

[63] Shashikumar A, Bičárová S, Laurence DR. The effect of ozone on pine forests in South-Eastern France from 2017 to 2019. J Forestry Res, 2023, 34(2301-315

[64] Shen YB, Peng Y, Zhu XC, Li HM, Zhang LW, Kong FL, Wang J, Yu D. The phytochemicals and health benefits of Cyclocarya paliurus (Batalin) Iljinskaja. Front Nutr, 2023, 10: 1158158

[65] Shi C, Kitao M, Agathokleous E, Watanabe M, Tobita H, Yazaki K, Kitaoka S, Koike T. Foliar chemical composition of two oak species grown in a free-air enrichment system with elevated O₃ and CO₂. J Agric Meteorol, 2016, 72(1): 50-58

[66] Shi C, Watanabe T, Koike T. Leaf stoichiometry of deciduous tree species in different soils exposed to free-air O₃ enrichment over two growing seasons. Environ Exp Bot, 2017, 138: 148-163

[67] Sicard P, Agathokleous E, De Marco A, Paoletti E. Ozone-reducing urban plants: choose carefully. Science, 2022, 377(6606585

[68] Singh AA, Ghosh A, Agrawal M, Agrawal SB. Secondary metabolites responses of plants exposed to ozone: an update. Environ Sci Pollut Res Int, 2023, 30(3888281-88312

[69] Skórka M, Sieprawska A, Telk A. The implication of manganese surplus on plant cell homeostasis: a review. J Plant Growth Regul, 2023, 42(3): 1327-1341

[70] Taib M, Rezzak Y, Bouyazza L, Lyoussi B. Medicinal uses, phytochemistry, and pharmacological activities of Quercus species. Evid Based Complement Altern Med, 2020, 2020: 1920683

[71] Tallmadge GK (1977) Ideabook: the joint dissemination review panel. U.S. Dept. of Health, Education and Welfare, National Institute of Education, U.S. Office of Education, Washington

[72] Thor K. Calcium-nutrient and messenger. Front. Plant Sci, 2019, 10: 440

[73] USDA (2024) Nuttall Oak [WWW Document]. https://www.srs.fs.usda.gov/pubs/misc/ag_654/volume_2/quercus/nutallii.htm

[74] Wallin G, Karlsson PE, Selldén G, Ottosson S, Medin EL, Pleijel H, Skärby L. Impact of four years exposure to different levels of ozone, phosphorus and drought on chlorophyll, mineral nutrients, and stem volume of Norway spruce. Picea Abies Physiol Plant, 2002, 114(2): 192-206

[75] Wang T, Xue LK, Feng ZZ, Dai JN, Zhang YN, Tan Y. Ground-level ozone pollution in China: a synthesis of recent findings on influencing factors and impacts. Environ Res Lett, 2022, 17(6 063003

[76] Wang YT, Wang YC, Feng ZZ, Yuan XY, Zhao Y. The impacts of ambient ozone pollution on China’s wheat yield and forest production from 2010 to 2021. Environ Pollut, 2023, 330 121726

[77] Wang C, Tang RJ, Kou SH, Xu XS, Lu Y, Rauscher K, Voelker A, Luan S. Mechanisms of calcium homeostasis orchestrate plant growth and immunity. Nature, 2024, 627(8003382-388

[78] Wolf FM. Meta-analysis: quantitative methods for research synthesis, 1986, Beverly Hills, Sage Publications72p

[79] Xia LL, Lam SK, Kiese R, Chen DL, Luo YQ, van Groenigen KJ, Ainsworth EA, Chen J, Liu SW, Ma L, Zhu YH, Butterbach-Bahl K. Elevated CO₂ negates O₃ impacts on terrestrial carbon and nitrogen cycles. One Earth, 2021, 4(12): 1752-1763

[80] Yan RS, Wang HL, Huang C, An JY, Bai HM, Wang Q, Gao YQ, Jing SG, Wang YY, Su H. Impact of spatial scales of control measures on the effectiveness of ozone pollution mitigation in Eastern China. Sci Total Environ, 2024, 906 167521

[81] Ye H, Li C, Ye W, Zeng F, Liu F, Liu Y, Wang F, Ye Y, Lin F, Li J. Ye H, Li C, Ye W, Zeng F. Medicinal angiosperms of Ulmaceae. Common Chinese Materia Medica: Volume 5, 2021, Singapore, Springer Nature 6186

[82] Yuan ZY, Chen HYH. Decoupling of nitrogen and phosphorus in terrestrial plants associated with global changes. Nat Clim Change, 2015, 5: 465-469

[83] Zhang YL, Han ZY, Li XY, Zhang HL, Yuan XY, Feng ZZ, Wang P, Mu ZB, Song W, Blake DR, Ying Q, George C, Sheng GY, Peng PA, Wang XM. Plants and related carbon cycling under elevated ground-level ozone: a mini review. Appl Geochem, 2022, 144 105400

[84] Zhang K, Xian L, Shang B, Xu YS, Feng ZZ, Agathokleous E. Fluctuating ozone exposures caused trade-offs between vegetative growth and reproduction of two Chinese bean cultivars and ethylenediurea alleviated ozone phytotoxicities. J Environ Sci in Press, 2025

[85] Zhou XY, Yue X, Tian CG. Responses of ecosystem productivity to anthropogenic ozone and aerosols at the 2060. Earth’s Futur, 2024, 12(1E2023EF003781

[86] Zhuang MH, Lam SK, Li YC, Chen SL. Elevated tropospheric ozone affects the concentration and allocation of mineral nutrients of two bamboo species. Sci Total Environ, 2017, 577: 231-235

RIGHTS & PERMISSIONS

The Author(s)