Effects of elevated O3 and CO2 on chlorophyll fluorescence and gas exchange in Scots pine during the third growing season

Naturally regenerated, 30-year-old Scots pines (Pinus Sylvestris L.) were grown in open-top chambers and exposed in situ to doubled ambient O(3), doubled ambient CO(2) and a combination of elevated O(3) and CO(2) from 15 April to 15 September for three growing seasons (1994-1996). To examine the effects of O(3) and/or CO(2) on photosynthesis, chlorophyll a fluorescence and gas exchange were measured simultaneously. Doubled ambient O(3) significantly decreased the rates of photosynthesis at all levels of photon flux density. This was related mainly to a significant decrease in the photochemical efficiency of photosystem II (PS II) and the rate of whole electron transport, rather than to a decrease in stomatal conductance. When measurements were made at doubled ambient concentration of CO(2) (700 micromol mol(-1)), doubled ambient CO(2) treatment did not lead to a significant change in the intrinsic capacity of photosynthesis, as manifested by no changes in PS II, the rate of electron transport, the maximal rate of photosynthesis and the apparent quantum yield of CO(2) assimilation. However, elevated CO(2) increased the sensitivity of stomatal conductance to light and decreased maximal stomatal conductance. When O(3) and CO(2) were combined, the O(3)-induced decrease in photosynthesis rate was reduced significantly by a high concentration of CO(2). This may be partly related to the decrease in stomatal conductance induced by the high concentration of CO(2). The complete mechanism behind this interaction is, however, still unclear.     

Differential response of CO2 uptake parameters of soil- and sand-grown phaseolus vulgaris (L.) plants to absorbed ozone flux

Shoots of a soil- or sand-grown dwarf bean variety were exposed to O(3) concentrations in the range of 500 to 900 ppb for up to 5 h. The measured exchange rates of water vapor and CO(2) during exposures were used to calculate stomatal and mesophyll conductances averaged across all leaves. Changes in conductances were related to exposure duration and absorbed O(3) totals (AOT). Both conductances were more sensitive to AOT in sand-grown plants, which also had more visible injury under comparable AOT values. Measurements of the relationship between CO(2) exchange and internal CO(2) concentration of single leaflets of treated plants also showed greater sensitivity of CO(2)-saturated photosynthesis in sand-grown plants. Diffusional processes were not likely to have been the cause of dissimilar responses because the O(3) absorption rate was lower in sand-grown plants. A difference in the scaveninng capacities in cells is suggested to be the cause of the differences in sensitivity to acute O(3) exposure.     

Does living in elevated CO2 ameliorate tree response to ozone? A review on stomatal responses

Short-term elevated O3 reduces photosynthesis, which reduces stomatal conductance (g(s)) in response to increased substomatal CO2 concentration (Ci). Further exposure causes stomata to become sluggish in response to environmental stimuli. Exposure to elevated CO2 stimulates rapid stomata closure in response to increased Ci. This reduction in g(s) may not be sustained over time as photosynthesis down-regulates and with it, g(s). The relationship between g(s) and photosynthesis may not be constant because stomata respond more slowly to environmental changes than photosynthesis, and because elevated CO2 may alter guard cell sensitivity to other signals. Also, reduced stomatal density (and g(s)) in response to long-term CO2 enrichment suggests sustained reduction in g(s). Elevated CO2 is believed to ameliorate the deleterious O3 effects by reducing g(s) and thus the potential O3 flux into leaves. Confirmation that g(s) acclimation to CO2 enrichment does not lessen over time is critical for developing meaningful O3 flux scenarios.     

Effects of elevated CO2 concentration on the polyamine levels of field-grown soybean at three O3 regimes

Effects of increased ozone (O3) and carbon dioxide (CO2) on polyamine levels were determined in soybean (Glycine max L. Merr. cv. Clark) grown in open-top field chambers. The chamber treatments consisted of three O3 regimes equal to charcoal filtered (CF), non-filtered (NF), and non-filtered plus 40 nl litre(-1) O3 and CO2 treatments equal to 350, 400 and 500 microl litre(-1) for a total of nine treatments. Leaf samples were taken at three different times during the growing season. Examination of growth and physiological characteristics, such as photosynthesis, stomatal resistance, and shoot weight, revealed that increasing CO2 ameliorated the deleterious effects of increased O3. Results from the initial harvest, at the pre-flowering growth stage (23 days of treatment), showed that increasing O3 at ambient CO2 caused increases in putrescine (Put) and spermidine (Spd) of up to six-fold. These effects were lessened with increased CO2. Elevated CO2 increased polyamines in plants treated with CF air, but had no effect in the presence of ambient or enhanced O3 levels. Leaves harvested during peak flowering (37 days of treatment) showed O3-induced increases in Put and Spd at ambient CO2 concentrations. However, increased CO2 levels inhibited this response by blocking the O3-induced polyamine increase. Leaves harvested during the pod fill stage (57 days of treatment) showed no significant O3 or CO2 effects on polyamine levels. Our results demonstrate that current ambient O3 levels induce the accumulation of Put and Spd early in the growing season and that further increases in O3 could result in even greater polyamine increases. These results are consistent with a possible antiozonant function for polyamines. The ability of increased CO2 to protect soybeans from O3 damage, however, does not appear to involve polyamine accumulation.     

Ozone-induced changes in CO2 assimilation, O2 evolution and chlorophyll a fluorescence transients in barley

Spring barley (Hordeum vulgare cv. Klaxon) plants, 9 days old, were exposed to 0.05, 0.10 or 0.15 microl litre(-1) ozone (O3) for 12 days. Fumigation was administered for 7 h between 9.00 h and 16.00 h each day. Using conventional IRGA equipment, the carbon dioxide exchange rate (CER) was shown to decrease with increasing concentration of O3 during the exposure period, falling to 60% of the control value at the highest O3 concentration. Transpiration rates and stomatal conductance showed similar trends. Light saturation curves, obtained using a leaf disc oxygen electrode, demonstrated that O3-treated leaves had lower apparent quantum yields (QY) and generally lower rates of O2 evolution at saturating light and CO2 levels. Oscillations in chlorophyll a fluorescence, normally observed in control plants, could not be detected after O3 treatment and could only be restored to some extent by feeding the phosphate sequestering agent D-mannose to the leaves.     

Interactive effects of CO2 and O3 on a ponderosa pine plant/litter/soil mesocosm.

To study individual and combined impacts of two important atmospheric trace gases, CO2 and O3, on C and N cycling in forest ecosystems; a multi-year experiment using a small-scale ponderosa pine (Pinus ponderosa Laws.) seedling/soil/litter system was initiated in April 1998. The experiment was conducted in outdoor, sun-lit chambers where aboveground and belowground ecological processes could be studied in detail. This paper describes the approach and methodology used, and presents preliminary data for the first two growing seasons. CO2 treatments were ambient and elevated (ambient + 280 ppm). O3 treatments were elevated (hourly averages to 159 ppb, cumulative exposure > 60 ppb O3, SUM 06 approximately 10.37 ppm h), and a low control level (nearly all hourly averages <40 ppb. SUM 06 approximately 0.07 ppm h). Significant (P < 0.05) individual and interactive effects occurred with elevated CO2 and elevated O3. Elevated CO2 increased needle-level net photosynthetic rates over both seasons. Following the first season, the highest photosynthetic rates were for trees which had previously received elevated O3 in addition to elevated CO2. Elevated CO2 increased seedling stem diameters, with the greatest increase at low O3. Elevated CO2 decreased current year needle % N in the summer. For 1-year-old needles measured in the fall there was a decrease in % N with elevated CO2 at low O3, but an increase in % N with elevated CO2 at elevated O3. Nitrogen fixation (measured by acetylene reduction) was low in ponderosa pine litter and there were no significant CO2 or O3 effects. Neither elevated CO2 nor elevated O3 affected standing root biomass or root length density. Elevated O3 decreased the % N in coarse-fine (1-2 mm diameter) but not in fine (< 1 mm diameter) roots. Both elevated CO2 and elevated O3 tended to increase the number of fungal colony forming units (CFUs) in the AC soil horizon, and elevated O3 tended to decrease bacterial CFUs in the C soil horizon. Thus, after two growing seasons we showed interactive effects of O3 and CO2 in combination, in addition to responses to CO2 or O3 alone for a ponderosa pine plant/litter/soil system.     

Effects of decadal exposure to interacting elevated CO2 and/or O3 on paper birch (Betula papyrifera) reproduction

We studied the effects of long-term exposure (nine years) of birch (Betula papyrifera) trees to elevated CO(2) and/or O(3) on reproduction and seedling development at the Aspen FACE (Free-Air Carbon Dioxide Enrichment) site in Rhinelander, WI. We found that elevated CO(2) increased both the number of trees that flowered and the quantity of flowers (260% increase in male flower production), increased seed weight, germination rate, and seedling vigor. Elevated O(3) also increased flowering but decreased seed weight and germination rate. In the combination treatment (elevated CO(2)+O(3)) seed weight is decreased (20% reduction) while germination rate was unaffected. The evidence from this study indicates that elevated CO(2) may have a largely positive impact on forest tree reproduction and regeneration while elevated O(3) will likely have a negative impact.     

Impacts of elevated CO2 and/or O3 on leaf ultrastructure of aspen (Populus tremuloides) and birch (Betula papyrifera) in the aspen FACE experiment.

Impacts of elevated atmospheric O3 and/or CO2 on three clones of aspen (Populus tremuloides Michx.) and birch (Betula papyrifera Marsh.) were studied to determine, whether or not elevated CO2 ameliorates O3-induced damage to leaf cells. The plants were exposed for 3 years at the Aspen FACE exposure site in Wisconsin (USA) prior to sampling for ultrastructural investigations on 19 June 1999. In the aspen clones, elevated CO2 increased chloroplast cover index, leaf and spongy mesophyll layer thickness, intercellular air space volume in mesophyll, amount of starch in chloroplasts and cytoplasmic lipids but decreased the number of plastoglobuli in chloroplasts. In contrast, elevated O3 decreased chloroplast cover index, starch content, and the proportion of cytoplasm and intercellular space in mesophyll, and increased the proportion of vacuoles, the amount of condensed vacuolar tannins and the number of plastoglobuli. Ozone also caused structural thylakoid injuries (dilation, distortion) and stromal condensation in chloroplasts, which was ameliorated by elevated CO2 by 5-66% in aspen clones and by 2-10% in birch. Birch ultrastructure was less affected by elevated CO2 or O3 stress compared to aspen. In the most O3-sensitive aspen clone, thinner leaves and cell walls, lower proportion of cell wall volume, and higher volume for vacuoles was found compared to more-tolerant clones.     

Effects of enhanced O3 and CO2 enrichment on plant characteristics in wheat and corn

The effects of CO(2) enrichment and O(3) induced stress on wheat (Triticum aestivum L.) and corn (Zea mays L.) were studied in field experiments using open-top chambers to simulate the atmospheric concentrations of these two gases that are predicted to occur during the coming century. The experiments were conducted at Beltsville, MD, during 1991 (wheat and corn) and 1992 (wheat). Crops were grown under charcoal filtered (CF) air or ambient air + 40 nl liter(-1) O(3) (7 h per day, 5 days per week) having ambient CO(2) concentration (350 microl liter(-1) CO(2)) or + 150 microl liter(-1) CO(2) (12 h per day.). Averaged over O(3) treatments, the CO(2)-enriched environment had a positive effect on wheat grain yield (26% in 1991 and 15% in 1992) and dry biomass (15% in 1991 and 9% in 1992). Averaged over CO(2) treatments, high O(3) exposure had a negative impact on wheat grain yield (-15% in 1991 and -11% in 1992) and dry biomass (-11% in 1991 and -9% in 1992). Averaged over CO(2) treatments, high O(3) exposure decreased corn grain yield by 9%. No significant interactive effects were observed for either crop. The results indicated that CO(2) enrichment had a beneficial effect in wheat (C(3) crop) but not in corn (C(4) crop). It is likely that the O(3)-induced stress will be diminished under increased atmospheric CO(2) concentrations; however, maximal benefits in crop production in wheat in response to CO(2) enrichment will not be materialized under concomitant increases in tropospheric O(3) concentration.     

Foliar antioxidant status of adult Mediterranean oak species (Quercus ilex L. and Q. pubescens Willd.) exposed to permanent CO2-enrichment and to seasonal water stress.

Foliar antioxidants were measured in adult individuals of holm oak (Quercus ilex L.) and white oak (Q. pubescens Willd.) growing in the field either within the vicinity of natural CO2 springs or at a nearby control site under ambient CO2, which had been previously exposed to either daily irrigation or no irrigation. In oak trees permanently exposed to elevated CO2 the activities of antioxidant enzymes tended to be lower and the ascorbate pool was larger and more in reduced form, suggesting an attenuation of the oxidative risk in the CO2-enriched trees. In the enriched individuals of both species. the imposition of water shortage significantly increased the size of the glutathione pool and the total superoxide dismutase activity in a species-specific manner. Moreover, water-stressed trees exposed to elevated CO2 tended to have higher catalase and ascorbate peroxidase activities than water-stressed control trees. Such changes may reflect the need for an enhanced compensatory effort when trees acclimated to elevated CO2 are exposed to oxidative stress-promoting environmental factors, such as water shortage.     

Canopy profiles of photosynthetic parameters under elevated CO2 and N fertilization in a poplar plantation

A poplar plantation has been exposed to an elevated CO2 concentration for 5 years using the free air CO2 enrichment (FACE) technique. Even after such a long period of exposure, leaves of Populus x euramericana have not shown clear signs of photosynthetic acclimation. Only at the end of the growing season for shade leaves was a decrease of maximum velocity of carboxylation (Vcmax) observed. Maximum electron transport rate (Jmax) was increased by FACE treatment in July. Assimilation rates at CO2 partial pressure of 400 (A400) and 600 (A600) micromol mol(-1) were not significantly different under FACE treatment. Most notably FACE significantly decreased stomatal conductance (g(s)) both on upper and lower canopy leaves. N fertilization increased N content in the leaves on mass basis (Nm) and specific leaf area (SLA) in both CO2 treatments but did not influence the photosynthetic parameters. These data show that in poplar plantations the long-term effects of elevated CO2 on photosynthesis do not differ considerably from the short-term ones even with N deposition.     

A statistical approach to estimate O3 uptake of ponderosa pine in a mediterranean climate

In highly polluted sites, stomatal behavior is sluggish with respect to light, vapor pressure deficit, and internal CO2 concentration (Ci) and poorly described by existing models. Statistical models were developed to estimate stomatal conductance (gs) of 40-year-old ponderosa pine at three sites differing in pollutant exposure for the purpose of calculating O3 uptake. Gs was estimated using julian day, hour of day, pre-dawn xylem potential and photosynthetic photon flux density (PPFD). The median difference between estimated and observed field gs did not exceed 10 mmol H2O m(-2) s(-1), and estimated gs within 95% confidence intervals. 03 uptake was calculated from hourly estimated gs, hourly O3 concentration, and a constant to correct for the difference in diffusivity between water vapor and 03. The simulation model TREGRO was also used to calculate the cumulative 03 uptake at all three sites. 03 uptake estimated by the statistical model was higher than that simulated by TREGRO because gas exchange rates were proportionally higher. O3 exposure and uptake were significantly correlated (r2>0.92), because O3 exposure and gs were highly correlated in both statistical and simulation models.     

Acquired changes in stomatal characteristics in response to ozone during plant growth and leaf development of bush beans (Phaseolus vulgaris L.) indicate phenotypic plasticity

Bush bean (Phaseolus vulgaris L.) lines 'S156' (O3-sensitive)/'R123' (O3-tolerant) and cultivars 'BBL 290' (O3-sensitive)/'BBL 274' (O3-tolerant) were used to study the effects of O3 on stomatal conductance (gs), density, and aperture size on leaf and pod surfaces with the objective of establishing links between the degree of plant sensitivity to O3 and plasticity of stomatal properties in response to O3. Studies in open-top chambers (OTCs) and in continuously stirred tank reactors (CSTRs) established a clear relationship between plant developmental stages, degrees of O3 sensitivity and gs: while 'S156' had higher gs rates than 'R123' earlier in development, similar differences between 'BBL 290' and 'BBL 274' were observed at later stages. Gs rates on the abaxial leaf surfaces of 'S156' and 'BBL 290', accompanied by low leaf temperatures, were significantly higher than their O3-tolerant counterparts. Exposure to O3 in CSTRs had greater and more consistent impacts on both stomatal densities and aperture sizes of O3-sensitive cultivars. Stomatal densities were highest on the abaxial leaf surfaces of 'S156' and 'BBL 290' at higher O3 concentrations (60 ppb), but the largest aperture sizes were recorded on the adaxial leaf surfaces at moderate O3 concentrations (30 ppb). Exposure to O3 eliminated aperture size differences on the adaxial leaf surfaces between sensitive and tolerant cultivars. Regardless of sensitivity to O3 and treatment regimes, the smallest aperture sizes and highest stomatal densities were found on the abaxial leaf surface. Our studies showed that O3 has the potential to affect stomatal plasticity and confirmed the presence of different control mechanisms for stomatal development on each leaf surface. This appeared to be more evident in O3-sensitive cultivars.     

Modelling stomatal responses of spring wheat (Triticum aestivum L. cv. Turbo) to ozone and different levels of water supply

Spring wheat (Triticum aestivum L. cv. Turbo) was exposed to different levels of ozone and water supply in open-top chambers in 1991. Air was charcoal filtered (CF), non-filtered (NF) and CF plus proportional addition of ambient or twice ambient ozone (CF1, CF2). Seasonal means of O(3), taken over 24 h, were 2.3, 20.6, 17.3, and 34.5 nl litre(-1) for CF, NF, CF1 and CF2 treatments, respectively. A split-plot design was used to obtain two levels of water supply: one-half of the pots was irrigated sufficiently not to show any symptoms of drought stress; the others were exposed to low water supply and received 50% of these amounts. Using a steady-state porometer approximately 800 measurements of stomatal conductance (g(s)) were made on flag leaves from 68 to 106 days after sowing. The measurements yielded only small differences of maximum conductance between the two levels of water supply. Therefore, low water supply did not protect wheat plants against ozone injury via reduced stomatal uptake in this experiment. To describe the effects of environmental variables on the stomatal behaviour, boundary-line analysis and non-linear regression analysis were used. Besides microclimatic parameters, the ozone dose of flag leaves was introduced as an independent variable affecting stomatal aperture. A well-defined boundary line for ozone dose was found, suggesting that increasing ozone dose caused stomatal closure in wheat flag leaves. But at high ozone doses, co-acting senescence seems also responsible for the decrease in stomatal conductance. A multiplicative boundary-line model was used to predict stomatal conductance from combinations of environmental variables. In the test carried out with the measurements of stomatal conductance, the model accounted only for 40% of the variation of g(s). Generalized stomatal response patterns of the herbaceous growth form, the dependence of the variables' age and ozone dose and the lack of an important factor influencing stomatal response (water status of the plant) in the model, are suggested as explanations of the poor results of the test.     

The ability of apoplastic ascorbate to protect poplar leaves against ambient ozone concentrations: a quantitative approach

Shoots of a sensitive (Populus nigra 'Brandaris') and a more tolerant (Populus euramericana 'Robusta') poplar clones were exposed for 30 days to Filtered Air or ambient O3-concentrations in fumigation cabinets. At regular intervals were determined: gas exchange of the leaves, the internal air space (Vair) and apoplastic water volume (Vapo) and the reduced (ASA) and oxidized (DHA) ascorbate concentration in the apoplast and in the mesophyll cells. The apoplastic ASA-concentration was 0.2 mM at the start of the experiment for both cultivars, while the effective cell wall thickness, estimated from Vapo, varied from 0.3 to 0.6 micron. Model calculations revealed that only 30% of the O3 molecules entering the apoplast was intercepted at these values. The O3-treatment induced a decline in stomatal conductance, an increase in Vapo and in the apoplastic ASA-concentration. As a result the estimated O3-flux to the cell membrane strongly declined. However, these responses occurred after the O3-induced reduction in photosynthesis. Moreover, they did not prevent early senescence of the leaves at a prolonged exposure. Therefore, it is concluded that the increase in apoplastic ASA-concentration was rather a general stress reaction of the affected poplar leaf than a (specific) defence reaction induced by O3. Our results suggest that other factors than the scavenging efficiency of apoplastic ASA were responsible for the difference in O3 sensitivity between both poplar cultivars.     

Consequences of elevated carbon dioxide and ozone for foliar chemical composition and dynamics in trembling aspen (Populus tremuloides) and paper birch (Betula papyrifera).

Atmospheric chemical composition affects foliar chemical composition, which in turn influences the dynamics of both herbivory and decomposition in ecosystems. We assessed the independent and interactive effects of CO2 and O3 fumigation on foliar chemistry of quaking aspen (Populus tremuloides) and paper birch (Betula papyrifera) at a Free-Air CO2 Enrichment (FACE) facility in northern Wisconsin. Leaf samples were collected at five time periods during a single growing season, and analyzed for nitrogen. starch and condensed tannin concentrations, nitrogen resorption efficiencies (NREs), and C:N ratios. Enriched CO2 reduced foliar nitrogen concentrations in aspen and birch; O3 only marginally reduced nitrogen concentrations. NREs were unaffected by pollution treatment in aspen, declined with 03 exposure in birch, and this decline was ameliorated by enriched CO2. C:N ratios of abscised leaves increased in response to enriched CO2 in both tree species. O3 did not significantly alter C:N ratios in aspen, although values tended to be higher in + CO2 + O3 leaves. For birch, O3 decreased C:N ratios under ambient CO2 and increased C:N ratios under elevated CO2. Thus, under the combined pollutants, the C:N ratios of both aspen and birch leaves were elevated above the averaged responses to the individual and independent trace gas treatments. Starch concentrations were largely unresponsive to CO2 and O3 treatments in aspen. but increased in response to elevated CO2 in birch. Levels of condensed tannins were negligibly affected by CO2 and O3 treatments in aspen, but increased in response to enriched CO2 in birch. Results from this work suggest that changes in foliar chemical composition elicited by enriched CO2 are likely to impact herbivory and decomposition, whereas the effects of O3 are likely to be minor, except in cases where they influence plant response to CO2.     

Impact of increased springtime O3 exposure on Scots pine (Pinus sylvestris) seedlings in central Finland

Three-year-old Scots pine (Pinus sylvestris L.) seedlings were exposed to ambient or elevated ozone (O(3)) concentrations in open-air exposure fields in central Finland in 1995-97. Three different treatments were applied in 1996 and 1997: ambient air, elevated O(3) (1.3-1.5xambient) during the growing season (June-September) and elevated O(3) in March-September, i.e. the growing season including the springtime O(3) exposure. The ambient mean O(3) concentrations were 40% higher in springtime (March-May) compared to the concentrations during the growing seasons. Maximum O(3) concentrations were measured in April or early May, whereas a clear increase in the stomatal activity of the seedlings was observed by the middle of May. This suggests a low intake of O(3) by conifers despite the higher O(3) concentrations in spring. Stomatal conductance, and contents of chlorophyll and ribulosebisphosphate carboxylase/oxygenase (Rubisco) in current-year needles were not significantly affected by any O(3) treatment. Only a slight decrease in current-year shoot growth, slight increase in the abscission of 2-year-old needles and increased electron density of chloroplast stroma by springtime O(3) exposure suggest a rather small contribution of elevated springtime O(3) concentrations to total O(3) damage under current climatic conditions in Finland. However, the increases in springtime O(3) concentrations may enhance the cumulative effects of O(3) during long-term O(3) exposures.     

Interactive effects of ozone and elevated carbon dioxide on the growth and physiology of black cherry, green ash, and yellow-poplar seedlings

Potted seedlings of black cherry (Prunus serotina Ehrh.) (BC), green ash (Fraxinus pennsylvanica Marsh.) (GA), and yellow-poplar (Liriodendron tulipifera L.) (YP) were exposed to one of the four treatments: (1) charcoal-filtered air (CF) at ambient CO(2) (control); (2) twice ambient O(3) (2 x O(3)); (3) twice ambient CO(2) (650 microl l(-1)) plus CF air (2 x CO(2)); or (4) twice ambient CO(2) (650 microl l(-1)) plus twice ambient O(3) (2 x CO(2) + 2 x O(3)). The treatments were duplicated in eight continuously stirred tank reactors for 10 weeks. Gas exchange was measured during the last 3 weeks of treatment and all seedlings were destructively harvested after 10 weeks. Significant interactive effects of O(3) and CO(2) on the gas exchange of all three species were limited. The effects of elevated CO(2) and O(3), singly and combined, on light-saturated net photosynthesis (A(max)) and stomatal conductance (g(s)) were inconsistent across species. In all three species, elevated O(3) had no effect on g(s). Elevated CO(2) significantly increased A(max) in GA and YP foliage, and decreased g(s) in YP foliage. Maximum carbon exchange rates and quantum efficiencies derived from light-response curves increased, while compensation irradiance and dark respiration decreased in all three species when exposed to 2 x CO(2). Elevated O(3) affected few of these parameters but any change that was observed was opposite to that from exposure to 2 x CO(2)-air. Interactive effects of CO(2) and O(3) on light-response parameters were limited. Carboxylation efficiencies, derived from CO(2)-response curves (A/C(i) curves) decreased only in YP foliage exposed to 2 x CO(2)-air. In general, growth was significantly stimulated by 2 x CO(2) in all three species; though there were few significant growth responses following exposure to 2 x O(3) or the combination of 2 x CO(2) plus 2 x O(3). Results indicate that responses to interacting stressors such as O(3) and CO(2) are species specific.     

Visible and microscopic injury in leaves of five deciduous tree species related to current critical ozone levels

Because the current critical level of ozone (O(3)) for forest trees is based only on one species, the responses of five deciduous tree species were differentiated in a climate chamber experiment. The number of symptomatic leaves per tree was significantly increased, and stomatal conductance was decreased under 50% ambient+30 nl l(-1) O(3) as compared to 'normal' senescence at 50% ambient [O(3)]. Species with a high stomatal conductance did not show earlier or more leaf injury symptoms. The additional 30 nl l(-1) O(3) induced specific pectinaceous cell wall protrusions, phenolic cell wall incrustations, tonoplast vesicles, and inhomogeneous, condensed/precipitated phenolic material in the vacuoles. Due to added O(3), cell senescence was accelerated with increased electron-density of the cytoplasm, and initial chloroplast degeneration. The slow degeneration process started in mesophyll cells, and expanded into epidermal and finally guard cells. Because of the large variance in biomass between individuals and species, the current critical level is supported by the assessment of visible leaf symptoms rather than growth reduction.     

Tree age dependence and within-canopy variation of leaf gas exchange and antioxidative defence in Fagus sylvatica under experimental free-air ozone exposure

We characterized leaf gas exchange and antioxidative defence of two-year-old seedlings and 60-year-old trees of Fagus sylvatica exposed to ambient (1 x O3) or two-fold ambient (2 x O3) O3 concentrations (maximum of 150 ppb) in a free-air canopy exposure system throughout the growing season. Decline in photosynthesis from sun-exposed to shaded conditions was more pronounced in adult than juvenile trees. Seedling leaves and leaves in the sun-exposed canopy had higher stomatal conductance and higher internal CO2 concentrations relative to leaves of adult trees and leaves in shaded conditions. There was a weak overall depression of photosynthesis in the 2 x O3 variants across age classes and canopy positions. Pigment and tocopherol concentrations of leaves were significantly affected by canopy position and tree age, whereas differences between 1 x O3 and 2 x O3 regimes were not observed. Glutathione concentrations were significantly increased under 2 x O3 across both age classes and canopy levels. Seedlings differed from adult trees in relevant physiological and biochemical traits in ozone response. The water-soluble antioxidative systems responded most sensitively to 2 x O3 without regard of tree age or canopy position.