Spring leaf flush in aspen (Populus tremuloides) clones is altered by long-term growth at elevated carbon dioxide and elevated ozone concentration

Early spring leaf out is important to the success of deciduous trees competing for light and space in dense forest plantation canopies. In this study, we investigated spring leaf flush and how long-term growth at elevated carbon dioxide concentration ([CO₂]) and elevated ozone concentration ([O₃]) altered leaf area index development in a closed Populus tremuloides (aspen) canopy. This work was done at the Aspen FACE experiment where aspen clones have been grown since 1997 in conditions simulating the [CO₂] and [O₃] predicted for ∼2050. The responses of two clones were compared during the first month of spring leaf out when CO₂ fumigation had begun, but O₃ fumigation had not. Trees in elevated [CO₂] plots showed a stimulation of leaf area index (36%), while trees in elevated [O₃] plots had lower leaf area index (−20%). While individual leaf area was not significantly affected by elevated [CO₂], the photosynthetic operating efficiency of aspen leaves was significantly improved (51%). There were no significant differences in the way that the two aspen clones responded to elevated [CO₂]; however, the two clones responded differently to long-term growth at elevated [O₃]. The O₃-sensitive clone, 42E, had reduced individual leaf area when grown at elevated [O₃] (−32%), while the tolerant clone, 216, had larger mature leaf area at elevated [O₃] (46%). These results indicate a clear difference between the two clones in their long-term response to elevated [O₃], which could affect competition between the clones, and result in altered genotypic composition in future atmospheric conditions.     

Exposure to elevated ozone levels differentially affects the antioxidant capacity and the redox homeostasis of two subtropical Phaseolus vulgaris L. varieties

Ozone (O₃) has become one of the most toxic air pollutants to plants worldwide. However, investigations on O₃ impacts on crops health and productivity in South America countries are still scarce. The present study analyzed the differences on the enzymatic and non-enzymatic antioxidant system in foliar tissue of two subtropical Phaseolus vulgaris varieties exposed to high O₃ concentration. Both varieties were negatively impacted by the pollutant, but the responses between each variety were quite distinct. Results revealed that Irai has higher constitutive levels of reactive oxygen species (ROS) and ascorbate (AsA) concentration, but lower total thiol levels and catalase immunocontent. In this variety catalase protein concentration was increased after O₃ exposure, indicating a better cellular capacity to reduce hydrogen peroxide. On the opposite, Fepagro 26-exposed plants increased ROS generation and AsA concentration, but had the levels of total thiol content and catalase protein unchanged. Furthermore, O₃ treatment reduced the levels of chlorophylls a and b, and the relationship analysis between the chlorophyll ratio (a/b) and protein concentration were positively correlated indicating that photosynthetic apparatus is compromised, and thus probably is the biomass acquisition on Fepagro 26. Differently, O₃ treatment of Irai did not affect chlorophylls a and b content, and loss on the protein content was lower. Altogether, these data suggest that early accumulation of ROS on Fepagro 26 are associated with an insufficient leaf antioxidant capacity, which leads to cell structure disruption and impairs the photosynthesis. Irai seems to be more tolerant to O₃ toxic effects than Fepagro 26, and the observed differences on O₃ sensitivity between the two varieties are apparently based on constitutive differences involved in the maintenance of intracellular redox homeostasis.     

Temporal processes that contribute to nonlinearity in vegetation responses to ozone exposure and dose

Ozone interacts with plant tissue through distinct temporal processes. Sequentially, plants are exposed to ambient O₃ that (1) moves through the leaf boundary layer, (2) is taken up into plant tissue primarily through stomata, and (3) undergoes chemical interaction within plant tissue, first by initiating alterations and then as part of plant detoxification and repair. In this paper, we discuss the linkage of the temporal variability of apoplastic ascorbate with the diurnal variability of defense mechanisms in plants and compare this variability with daily maximum O₃ concentration and diurnal uptake and entry of O₃ into the plant through stomata. We describe the quantitative evidence on temporal variability in concentration and uptake and find that the time incidence for maximum defense does not necessarily match diurnal patterns for maximum O₃ concentration or maximum uptake. We suggest that the observed out-of-phase association of the diurnal patterns for the above three processes produces a nonlinear relationship that results in a greater response from the higher hourly average O₃ concentrations than from the lower or mid-level values. The fact that these out-of-phase processes affect the relationship between O₃ exposure/dose and vegetation effects ultimately impact the ability of flux-based indices to predict vegetation effects accurately for purposes of standard setting and critical levels. Based on the quantitative aspect of temporal variability identified in this paper, we suggest that the inclusion of a diurnal pattern for detoxification in effective flux-based models would improve the predictive characteristics of the models. While much of the current information has been obtained using high O₃ exposures, future research results derived from laboratory biochemical experiments that use short but elevated O₃ exposures should be combined with experimental results that use ambient-type exposures over longer periods of time. It is anticipated that improved understanding will come from future research focused on diurnal variability in plant defense mechanisms and their relationship to the diurnal variability in ambient O₃ concentration and stomatal conductance. This should result in more reliable O₃ exposure standards and critical levels.     

Intraspecific responses of six Indian clover cultivars under ambient and elevated levels of ozone

Intraspecific variation in six cultivars of clover Trifolium alexandrinum L., (Bundel, Wardan, JHB-146, Saidi, Fahli, and Mescavi) has been studied with ambient and elevated O₃ (ambient?+?10 ppb O₃) in open top chambers. Significant effect of elevated O₃ was detected on different morphological, physiological, and biochemical parameters depicting differential response among the test cultivars. Results showed that the magnitude of O₃ induced foliar injury symptoms varied in all the cultivars. Ozone significantly depressed photosynthetic rate, stomatal conductance, and photosynthetic efficiency, although variations were cultivar specific. Ozone treatment diminished total biomass of all the cultivars; reduction was highest in Wardan with least O₃ resistance followed by Bundel, JHB-146, Saidi, Mescavi, and Fahli. According to the cumulative sensitive index, variations in the sensitivity showed that two cultivars (Wardan and Bundel) were sensitive to elevated O₃, while other three cultivars (Fahli, Saidi, and Mescavi) were resistant, and JHB-146 showed intermediate sensitivity. Therefore, the present study supported the selection of sensitive cultivar of clover as a bioindicator for O₃ under Indian conditions for the areas experiencing higher concentrations of O₃.     

The annual variation in stomatal ammonia compensation point of rye grass (Lolium perenne L.) leaves in an intensively managed grassland

The stomatal ammonia compensation point for ammonia (NH₃) of an intensively managed pasture of rye grass (Lolium perenne L.) was followed from mid January till November 2000. Leaf samples were taken every week. Simultaneously, the ambient NH₃ concentration was measured. Meteorological data (temperature, wind speed, rainfall and radiance) were collected from a nearby field station. The vacuum infiltration technique was used to isolate the apoplastic solution of the leaves. From the determined ammonium (NH₄⁺) concentration and pH in the apoplast, the gaseous NH₃ concentration inside the leaves was calculated, i.e. the so-called stomatal compensation point (χ_s).Temperature appeared to have a predominant effect on χ_s, partly by affecting the equilibrium between gaseous NH₃ inside the leaf and NH₃ dissolved in the apoplast and partly by affecting physiological processes influencing the NH₄⁺ concentration in the apoplast. Results of the present study suggest that these temperature effects were counteracting. On one hand temperature increase during early spring stimulated NH₃ volatilisation from the apoplast, on the other hand it led to a decline in apoplastic NH₄⁺ from 0.9 to 0.2 mM, thereby diminishing the emission potential of the leaf. The low NH₄⁺ concentrations during spring and summer coincided with a low total leaf N content (<3% dw). However, there was no clear relationship between these two variables. The total N content of the leaf tissue is therefore an inadequate parameter for prediction of the potential NH₃ emission from rye grass leaves. No annual trend was found for the apoplast pH. With a few exceptions, pH varied between 5.9 and 6.5 throughout the experimental period.The calculated values for χ_s varied between 0.5 and 4 μg m⁻³. The gaseous NH₃ concentrations inside the grass leaves were, with a few exceptions, always smaller than the measured ambient NH₃ concentrations. The present study indicates that under the current ambient NH₃ concentrations in the Netherlands, the grass canopy is unlikely to be a major source of NH₃ emission.     

Stomatal uptake of O3 in aspen and aspen-birch forests under free-air CO2 and O3 enrichment

Rising atmospheric carbon dioxide (CO₂) may alleviate the toxicological impacts of concurrently rising tropospheric ozone (O₃) during the present century if higher CO₂ is accompanied by lower stomatal conductance (g_s), as assumed by many models. We investigated how elevated concentrations of CO₂ and O₃, alone and in combination, affected the accumulated stomatal flux of O₃ (AFst) by canopies and sun leaves in closed aspen and aspen-birch forests in the free-air CO₂-O₃ enrichment experiment near Rhinelander, Wisconsin. Stomatal conductance for O₃ was derived from sap flux data and AFst was estimated either neglecting or accounting for the potential influence of non-stomatal leaf surface O₃ deposition. Leaf-level AFst (AFst_l) was not reduced by elevated CO₂. Instead, there was a significant CO₂ × O₃ interaction on AFst_l, as a consequence of lower values of g_s in control plots and the combination treatment than in the two single-gas treatments. In addition, aspen leaves had higher AFst_l than birch leaves, and estimates of AFst_l were not very sensitive to non-stomatal leaf surface O₃ deposition. Our results suggest that model projections of large CO₂-induced reductions in g_s alleviating the adverse effect of rising tropospheric O₃ may not be reasonable for northern hardwood forests.     

Detoxification and repair process of ozone injury: from O3 uptake to gene expression adjustment

Plants react to O₃ threat by setting up a variety of defensive strategies involving the co-ordinated modulation of stress perception, signalling and metabolic responses. Although stomata largely controls O₃ uptake, differences in O₃ tolerance cannot always be ascribed to changes in stomatal conductance but cell protective and repair processes should be taken into account. O₃-driven ROS production in the apoplast induces a secondary, active, self-propagating generation of ROS, whose levels must be finely tuned, by many enzymatic and non-enzymatic antioxidant systems, to induce gene activation without determining uncontrolled cell death. Additional signalling molecules, as ethylene, jasmonic and salicylic acid are also crucial to determine the spreading and the containment of leaf lesions. The main recent results obtained on O₃ sensing, signal transduction, ROS formation and detoxification mechanisms are here discussed.     

Effects of chronic elevated ozone exposure on gas exchange responses of adult beech trees (Fagus sylvatica) as related to the within-canopy light gradient

The effects of elevated O₃ on photosynthetic properties in adult beech trees (Fagus sylvatica) were investigated in relation to leaf mass per area as a measure of the gradually changing, within-canopy light availability. Leaves under elevated O₃ showed decreased stomatal conductance at unchanged carboxylation capacity of Rubisco, which was consistent with enhanced δ¹³C of leaf organic matter, regardless of the light environment during growth. In parallel, increased energy demand for O₃ detoxification and repair was suggested under elevated O₃ owing to enhanced dark respiration. Only in shade-grown leaves, light-limited photosynthesis was reduced under elevated O₃, this effect being accompanied by lowered F_v/F_m. These results suggest that chronic O₃ exposure primarily caused stomatal closure to adult beech trees in the field regardless of the within-canopy light gradient. However, light limitation apparently raised the O₃ sensitivity of photosynthesis and accelerated senescence in shade leaves.     

Elevated CO2 response of photosynthesis depends on ozone concentration in aspen

The effect of elevated CO₂ and O₃ on apparent quantum yield (?), maximum photosynthesis (P_max), carboxylation efficiency (V_cmax) and electron transport capacity (J_max) at different canopy locations was studied in two aspen (Populus tremuloides) clones of contrasting O₃ tolerance. Local light climate at every leaf was characterized as fraction of above-canopy photosynthetic photon flux density (%PPFD). Elevated CO₂ alone did not affect ? or P_max, and increased J_max in the O₃-sensitive, but not in the O₃-tolerant clone. Elevated O₃ decreased leaf chlorophyll content and all photosynthetic parameters, particularly in the lower canopy, and the negative impact of O₃ increased through time. Significant interaction effect, whereby the negative impact of elevated O₃ was exaggerated by elevated CO₂ was seen in Chl, N and J_max, and occurred in both O₃-tolerant and O₃-sensitive clones. The clonal differences in the level of CO₂ × O₃ interaction suggest a relationship between photosynthetic acclimation and background O₃ concentration.     

Assessment of competitive ability of two Indian wheat cultivars under ambient O3 at different developmental stages

The concentrations of O₃ are increasing, which may have potential adverse effects on crop yield. This paper deals with assessing the intraspecific variability of two wheat cultivars (PBW 343 and M 533) at different growth stages using open top chambers. Mean O₃ concentrations were 50.2 and 53.2 ppb, and AOT40 values were 9 and 12.1 ppm h, respectively, in 2008–2009 and 2009–2010. Reproductive stage showed higher AOT40 values (6.9 and 9.2 ppm h) compared to vegetative (2.23 and 2.9 ppm h). Critical levels of a 3-month AOT 40 of 3 ppm h led to 6 % yield reduction in two wheat cultivars for two consecutive years. Variations in photosynthesis rate, stomatal conductance (g_s), Fv/Fm ratio, photosynthetic pigments, primary and secondary metabolites, morphological parameters, and yield attributes were measured at vegetative and reproductive stages. Reductions in number of leaves, leaf area, total biomass, root/shoot ratio, RGR, photosynthetic pigments, protein content, and Fv/Fm ratio in PBW 343 were more than M 533 at reproductive stage. Photosynthetic rate did not vary between the cultivars, but g_s was higher in PBW 343 compared to M 533 under ambient O₃. Higher total phenolics and peroxidase activity were recorded in M 533 at reproductive stage conferring higher resistance at latter age. Results of O₃ resistance showed that M 533 was sensitive compared to PBW 343 during vegetative stage but developed more resistance at reproductive stage. PBW 343 with larger leaf area and high g_s is more sensitive than M 533 with smaller leaf area and low g_s. The study suggests that the sensitivity varied with plant growth stage, and the plant showing higher sensitivity during vegetative period developed more resistance during reproductive period due to higher defense mechanism. Though the yield reductions were same in both cultivars under ambient O₃, the mechanism of acquiring the resistance is different between the cultivars.     

Seasonal profiles of leaf ascorbic acid content and redox state in ozone-sensitive wildflowers

Cutleaf coneflower (Rudbeckia laciniata L.), crown-beard (Verbesina occidentalis Walt.), and tall milkweed (Asclepias exaltata L.) are wildflower species native to Great Smoky Mountains National Park (U.S.A.). Natural populations of each species were analyzed for leaf ascorbic acid (AA) and dehydroascorbic acid (DHA) to assess the role of ascorbate in protecting the plants from ozone stress. Tall milkweed contained greater quantities of AA (7-10 micromol g(-1) fresh weight) than crown-beard (2-4 micromol g(-1) fresh weight) or cutleaf coneflower (0.5-2 micromol g(-1) fresh weight). DHA was elevated in crown-beard and cutleaf coneflower relative to tall milkweed suggesting a diminished capacity for converting DHA into AA. Tall milkweed accumulated AA in the leaf apoplast (30-100 nmol g(-1) fresh weight) with individuals expressing ozone foliar injury symptoms late in the season having less apoplast AA. In contrast, AA was not present in the leaf apoplast of either crown-beard or cutleaf coneflower. Unidentified antioxidant compounds were present in the leaf apoplast of all three species. Overall, distinct differences in antioxidant metabolism were found in the wildflower species that corresponded with differences in ozone sensitivity.     

DNA damage in Populus tremuloides clones exposed to elevated O3

The effects of elevated concentrations of atmospheric tropospheric ozone (O₃) on DNA damage in five trembling aspen (Populus tremuloides Michx.) clones growing in a free-air enrichment experiment in the presence and absence of elevated concentrations of carbon dioxide (CO₂) were examined. Growing season mean hourly O₃ concentrations were 36.3 and 47.3 ppb for ambient and elevated O₃ plots, respectively. The 4th highest daily maximum 8-h ambient and elevated O₃ concentrations were 79 and 89 ppb, respectively. Elevated CO₂ averaged 524 ppm (+150 ppm) over the growing season. Exposure to O₃ and CO₂ in combination with O₃ increased DNA damage levels above background as measured by the comet assay. Ozone-tolerant clones 271 and 8L showed the highest levels of DNA damage under elevated O₃ compared with ambient air; whereas less tolerant clone 216 and sensitive clones 42E and 259 had comparably lower levels of DNA damage with no significant differences between elevated O₃ and ambient air. Clone 8L was demonstrated to have the highest level of excision DNA repair. In addition, clone 271 had the highest level of oxidative damage as measured by lipid peroxidation. The results suggest that variation in cellular responses to DNA damage between aspen clones may contribute to O₃ tolerance or sensitivity.     

Leaf size and surface characteristics of Betula papyrifera exposed to elevated CO2 and O3

Betula papyrifera trees were exposed to elevated concentrations of CO₂ (1.4 × ambient), O₃ (1.2 × ambient) or CO₂ + O₃ at the Aspen Free-air CO₂ Enrichment Experiment. The treatment effects on leaf surface characteristics were studied after nine years of tree exposure. CO₂ and O₃ increased epidermal cell size and reduced epidermal cell density but leaf size was not altered. Stomatal density remained unaffected, but stomatal index increased under elevated CO₂. Cuticular ridges and epicuticular wax crystallites were less evident under CO₂ and CO₂ + O₃. The increase in amorphous deposits, particularly under CO₂ + O_3, was associated with the appearance of elongated plate crystallites in stomatal chambers. Increased proportions of alkyl esters resulted from increased esterification of fatty acids and alcohols under elevated CO₂ + O₃. The combination of elevated CO₂ and O₃ resulted in different responses than expected under exposure to CO₂ or O₃ alone.     

Stomatal characteristics and infection biology of Pyrenopeziza betulicola in Betula pendula trees grown under elevated CO2 and O3

Two silver birch clones were exposed to ambient and elevated concentrations of CO₂ and O₃, and their combination for 3 years, using open-top chambers. We evaluated the effects of elevated CO₂ and O₃ on stomatal conductance (g_s), density (SD) and index (SI), length of the guard cells, and epidermal cell size and number, with respect to crown position and leaf type. The relationship between the infection biology of the fungus (Pyrenopeziza betulicola) causing leaf spot disease and stomatal characteristics was also studied. Leaf type was an important determinant of O₃ response in silver birch, while crown position and clone played only a minor role. Elevated CO₂ reduced the g_s, but had otherwise no significant effect on the parameters studied. No significant interactions between elevated CO₂ and O₃ were found. The infection biology of P. betulicola was not correlated with SD or g_s, but it did occasionally correlate positively with the length of the guard cells.     

Are Bavarian Forests (southern Germany) at risk from ground-level ozone? Assessment using exposure and flux based ozone indices

Exposure and flux-based indices of O₃ risk were compared, at 19 forest locations across Bavaria in southern Germany from 2002 to 2005; leaf symptoms on mature beech trees found at these locations were also examined for O₃ injury. O₃ flux modelling was performed using continuously recorded O₃ concentrations in combination with meteorological and soil moisture data collected from Level II forest sites. O₃ measurements at nearby rural open-field sites proved appropriate as surrogates in cases where O₃ data were lacking at forest sites (with altitude-dependent average differences of about 10% between O₃ concentrations). Operational thresholds of biomass loss for both O₃ indices were exceeded at the majority of the forest locations, suggesting similar risk under long-term average climate conditions. However, exposure-based indices estimated higher O₃ risk during dry years as compared to the flux-based approach. In comparison, minor O₃-like leaf injury symptoms were detected only at a few of the forest sites investigated. Relationships between flux-based risk thresholds and tree response need to be established for mature forest stands for validation of predicted growth reductions under the prevailing O₃ regimes.     

Use of Plants for Air Pollution Monitoring

Leaf injury data from acute and chronic exposure studies of Dare soybean were regressed against the logarithms of exposure time and O₃ and SO₂ concentrations to develop a new two-pollutant leaf injury model (which explains 88% of the variance) and to calculate the parameters of best fit for this new model and a previously developed one-pollutant model. Using the calculated parameters, the percentage of leaf surface Injured over a growing season by O₃, SO₂, or both simultaneously was estimated for an ambient air sampling site located 2 miles from a coal burning power plant. For this site, the one- and two-pollutant models predicted that SO₂ effects would be negligible If SO₂ concentrations never exceeded the National Ambient Air Quality Standard (NAAQS) of 0.50 ppm, averaged over 3 h. However, calculations suggest that O₃ may injure up to 24% of Dare soybean leaf surface over a growing season even though the O₃ NAAQS of 0.12 ppm, averaged over 1 h, is never exceeded. Because the 3 h SO₂ standard is exceeded at very few places, the O₃ model is usually sufficient to estimate Dare soybean leaf Injury. Leaf injury is estimated by taking the logarithm of the summation of each daytime hour’s exponentiated O₃ concentration (c) measured at an ambient air sampling site over a growing season. This is expressed as: z = -0.0828 + 0.4876 in (Σco₃ ^2.618), where z is the Gaussian transform of percent leaf injury. The methods developed in this paper, using Dare soybean data as an example, may apply to other plants.     

Use of the antiozonant ethylenediurea (EDU) in Italy: verification of the effects of ambient ozone on crop plants and trees and investigation of EDU's mode of action

Twenty-four experiments where EDU was used to protect plants from ozone (O₃) in Italy are reviewed. Doses of 150 and 450 ppm EDU at 2-3 week intervals were successfully applied to alleviate O₃-caused visible injury and growth reductions in crop and forest species respectively. EDU was mainly applied as soil drench to crops and by stem injection or infusion into trees. Visible injury was delayed and reduced but not completely. In investigations on mode of action, EDU was quickly (<2 h) uptaken and translocated to the leaf apoplast where it persisted long (>8 days), as it cannot move via phloem. EDU did not enter cells, suggesting it does not directly affect cell metabolism. EDU delayed senescence, did not affect photosynthesis and foliar nitrogen content, and stimulated antioxidant responses to O₃ exposure. Preliminary results suggest developing an effective soil application method for forest trees is warranted.     

Elevated ozone phytotoxicity ameliorations in mung bean {Vigna radiata (L.) Wilczek} by foliar nebulization of silicic acid and ascorbic acid

The present work provides an insight into the development of biochemical adaptations in mung beans against ozone (O₃) toxicity. The study aims to explore the O₃ stress tolerance potential of mung bean genotypes under exogenous application of growth regulators. The seeds of twelve mung bean genotypes were grown in plastic pots under controlled conditions in the glasshouse. Six treatments, control (ambient ozone level 40–45 ppb), ambient O₃ with ascorbic acid, ambient ozone with silicic acid, elevated ozone (120 ppb), elevated O₃ with ascorbic acid (10 mM), and elevated ozone with silicic acid (0.1 mM) were applied. The O₃ fumigation was carried out using an O₃ generator. The results revealed that ascorbic acid and silicic acid application decreased the number of plants with foliar O₃ injury symptoms in different degrees, i.e., zero, first, second, third, and fourth degrees; whereas 0–4 degree symptoms represent, no symptoms, symptoms occupying?<?1/4, 1/4–1/2, 1/2–3/4, and?>?3/4 of the total foliage area, respectively. Application of ascorbic acid and silicic acid also prevented the plants from the negative effects of O₃ in terms of fresh as well as dry matter production, leaf chlorophyll, carotenoids, soluble proteins and ascorbic acid, proline, and malondialdehyde (MDA) contents. Overall, silicic acid application proved more effective in reducing the negative effects of O₃ on mung bean genotypes as compared to that of the ascorbic acid. Three mung bean genotypes (NM 20–21, NM-2006, and NM-2016) were identified to have a better adaptive mechanism for O₃ toxicity tolerance and may be good candidates for future variety development programs.

     

Will photosynthetic capacity of aspen trees acclimate after long-term exposure to elevated CO2 and O3?

Photosynthetic acclimation under elevated carbon dioxide (CO₂) and/or ozone (O₃) has been the topic of discussion in many papers recently. We examined whether or not aspen plants grown under elevated CO₂ and/or O₃ will acclimate after 11 years of exposure at the Aspen Face site in Rhinelander, WI, USA. We studied diurnal patterns of instantaneous photosynthetic measurements as well as A/C_i measurements monthly during the 2004-2008 growing seasons. Our results suggest that the responses of two aspen clones differing in O₃ sensitivity showed no evidence of photosynthetic and stomatal acclimation under either elevated CO₂, O₃ or CO₂ + O₃. Both clones 42E and 271 did not show photosynthetic nor stomatal acclimation under elevated CO₂ and O₃ after a decade of exposure. We found that the degree of increase or decrease in the photosynthesis and stomatal conductance varied significantly from day to day and from one season to another.     

Inter- and intra-specific responses to elevated ozone and chamber climate in northern birches

We studied the responses of micropropagated, northern provenances of downy, mountain and silver birches to elevated ozone (O₃) and changing climate using open-top chambers (OTCs). Contrary to our hypothesis, northern birches were sensitive to O₃, i.e. O₃ levels of 31-36 ppb reduced the leaf and root biomasses by −10%, whereas wood biomass was affected to a lesser extent. The warmer and drier OTC climate enhanced growth in general, though there were differences among the species and clones, e.g. in bud burst and biomass production. Inter- and intra-specific responses to O₃ and changing climate relate to traits such as allocation patterns between the above- and belowground parts (i.e. root/shoot ratio), which further relate to nutrient and water economy. Our experiments may have mimicked future conditions quite well, but only long-term field studies can yield the information needed to forecast responses at both tree and ecosystem levels.