Nutrition and the ozone sensitivity of birch (Betula pendula)

 Cuttings of a single birch clone (Betula pendula) were grown in field fumigation chambers throughout the growing season in either filtered air (control) or 90/40 nl O₃ l^–1 (day/night). Both regimes were split into plants under high and low nutrient supply (macro- and micronutrients). The stomatal density of leaves was increased by ozone but was lowered at high nutrition, while the inner air space was hardly affected by the treatments. Ozone induced macroscopic leaf injury regardless of nutrition, but leaf shedding was delayed in the low-fertilized plants, despite O₃ uptake being similar to that in high-fertilized plants. The leaf turn-over was enhanced in the O₃-exposed high-fertilized plants, but length growth and leaf formation of stems were not affected by ozone in either nutrient regime. Leaves of high-fertilized plants showed O₃-caused decline in photosynthetic capacity, water-use efficiency, apparent carbon uptake efficiency and quantum yield earlier as compared with low-fertilized plants, whereas chlorophyll fluorescence (F_V/F_M) and leaf nitrogen concentration were rather stable. CO₂ uptake rate and rubisco activity of young leaves compensated for the O₃ injury in the ageing leaves of the low-fertilized plants. In 8-week-old leaves, however, the O₃-induced decline in CO₂ uptake did not differ between the nutrient regimes and was associated with increased dark respiration rather than changed photorespiration. The balance between CO₂ supply and demand was lost, as was stomatal limitation on CO₂ uptake. High nutrition did not help leaves to maintain a high photosynthetic capacity and life span under O₃ stress. Received: 6 July 1996 / Accepted: 4 June 1997     

Ozone sensitivity in Triticum durum and T. aestivum with respect to leaf injury, photosynthetic activity and free radical content

Sensitivity to ozone is highly variable in cultivars of different wheat species, leading to differences in leaf injury and yield. Not much is known about the physiological background of these differences. The objective of this study was to compare the effects of ozone on photosynthetic parameters in Triticum aestivum L. (spring wheat cv. Nandu, winter wheat cv. Perlo) and Triticum durum Desf. (cv. Extradur). Plants cultivated in pots were exposed to 80 nmol mol^?1 ozone, or were used as control plants in a greenhouse. Stages of growth and senescence of single leaves were recorded. Light-saturated net photosynthesis, leaf conductance for water vapour, and chlorophyll fluorescence were measured. Stomatal limitation was calculated from CO₂ response curves, and the free radical content of whole leaves measured by EPR spectroscopy. Senescence of single leaves was enhanced by the ozone-treatment in all three cultivars, in the order Nandu > Perlo > Extradur. Development of whole plants was slightly delayed in Perlo and Nandu, but was accelerated significantly in Extradur. The rate of net photosynthesis under light saturation (A_sat) decreased significantly in older, ozone-fumigated leaves of Perlo and Nandu but not of Extradur. Leaf conductance (g₁) showed a similar behaviour, but stomatal limitation (l) was similar between ozone-treated and control plants. Thus, an ozone-induced closure of stomata was not the reason for the observed difference in A_sat. Perlo and Nandu showed a significant, only partly reversible decrease in F_v/F_m in ozone-fumigated leaves, whereas in Extradur the decrease was fully reversible only in older leaves. Whole leaves of Extradur, in contrast to Perlo and Nandu, showed no increase in EPR free radical signals. The higher ozone tolerance of Extradur was thus not caused by decreased ozone uptake via the stomata, but by a better ability of photosynthetically active mesophyll cells to cope with photooxidative stress.     

Effects of CO(2) Enrichment and Carbohydrate Content on the Dark Respiration of Soybeans

During the period of most active leaf expansion, the foliar dark respiration rate of soybeans (Glycine max cv Williams), grown for 2 weeks in 1000 microliters CO₂ per liter air, was 1.45 milligrams CO₂ evolved per hour leaf density thickness, and this was twice the rate displayed by leaves of control plants (350 microliters CO₂ per liter air). There was a higher foliar nonstructural carbohydrate level (e.g. sucrose and starch) in the CO₂ enriched compared with CO₂ normal plants. For example, leaves of enriched plants displayed levels of nonstructural carbohydrate equivalent to 174 milligrams glucose per gram dry weight compared to the 84 milligrams glucose per gram dry weight found in control plant leaves. As the leaves of CO₂ enriched plants approached full expansion, both the foliar respiration rate and carbohydrate content of the CO₂ enriched leaves decreased until they were equivalent with those same parameters in the leaves of control plants. A strong positive correlation between respiration rate and carbohydrate content was seen in high CO₂ adapted plants, but not in the control plants.

Mitochondria, isolated simultaneously from the leaves of CO₂ enriched and control plants, showed no difference in NADH or malate-glutamate dependent O₂ uptake, and there were no observed differences in the specific activities of NAD⁺ linked isocitrate dehydrogenase and cytochrome c oxidase. Since the mitochondrial O₂ uptake and total enzyme activities were not greater in young enriched leaves, the increase in leaf respiration rate was not caused by metabolic adaptations in the leaf mitochondria as a response to long term CO₂ enrichment. It was concluded, that the higher respiration rate in the enriched plant's foliage was attributable, in part, to a higher carbohydrate status.

     

Effects of ozone on wild type and transgenic tobacco

Tocopherol cyclase (TC, encoded by gene VTE1) catalyzes the penultimate step of tocopherol synthesis. In this study we used wild type and transgenic tobacco plants overexpressing VTE1 from Arabidopsis to examine the role of tocopherol in ozone sensitivity. Wild type plants responded to an 4-h exposure to 300 nmol mol⁻¹ ozone by severe leaf necrosis while the transgenic lines exhibited limited injury. Compared with the wild type, VTE1-overexpressing plants had lower increase in hydrogen peroxide, malondialdehyde contents and ion leakage, and lower decrease of net photosynthetic rate 48 h following the ozone exposure. Transgenic plants also better maintained the structural integrity of the photosynthetic apparatus.     

Effects of CO 2 enrichment on photosynthesis,growth, and nitrogen metabolism of the seagrass Zostera noltii

Seagrass ecosystems are expected to benefit from the global increase in CO ₂ in the ocean because the photosynthetic rate of these plants may be C_i‐limited at the current CO ₂ level. As well, it is expected that lower external pH will facilitate the nitrate uptake of seagrasses if nitrate is cotransported with H⁺ across the membrane as in terrestrial plants. Here, we investigate the effects of CO ₂ enrichment on both carbon and nitrogen metabolism of the seagrass Zostera noltii in a mesocosm experiment where plants were exposed for 5 months to two experimental CO ₂ concentrations (360 and 700 ppm). Both the maximum photosynthetic rate (P_m) and photosynthetic efficiency (α) were higher (1.3‐ and 4.1‐fold, respectively) in plants exposed to CO ₂‐enriched conditions. On the other hand, no significant effects of CO ₂ enrichment on leaf growth rates were observed, probably due to nitrogen limitation as revealed by the low nitrogen content of leaves. The leaf ammonium uptake rate and glutamine synthetase activity were not significantly affected by increased CO ₂ concentrations. On the other hand, the leaf nitrate uptake rate of plants exposed to CO ₂‐enriched conditions was fourfold lower than the uptake of plants exposed to current CO ₂ level, suggesting that in the seagrass Z. noltii nitrate is not cotransported with H⁺ as in terrestrial plants. In contrast, the activity of nitrate reductase was threefold higher in plant leaves grown at high‐CO ₂ concentrations. Our results suggest that the global effects of CO ₂ on seagrass production may be spatially heterogeneous and depend on the specific nitrogen availability of each system. Under a CO ₂ increase scenario, the natural levels of nutrients will probably become limiting for Z. noltii. This potential limitation becomes more relevant because the expected positive effect of CO ₂ increase on nitrate uptake rate was not confirmed.     

Ozone uptake and its effect on photosynthetic parameters of two tobacco cultivars with contrasting ozone sensitivity

Leaf discs of the ozone tolerant tobacco (Nicotiana tabacum L.) cv. Bel B and of the ozone sensitive cv. Bel W3, were exposed to an acute ozone fumigation (300 ppb) for 3 h. We measured ozone uptake by leaves and physiological characteristics before, during and after the treatment, in order to determine if the different O₃ sensitivity was correlated to the leaf uptake. In the tolerant cv. Bel B, O₃ uptake was high during the first 2 h of ozone exposure and then decreased. In the sensitive cv. Bel W3, the rate of O₃ uptake decreased constantly during ozone fumigation. The estimated cumulative uptake over the treatment time was higher (200 ± 30 μmol m^–2) in Bel B than in Bel W3 (130 ± 12 μmol m^–2). Thus, the ozone sensitivity was not correlated with ozone uptake. Stomatal conductance and photosynthesis were significantly inhibited during the fumigation in both cultivars. However, these reductions were strong and irreversible in the cv. Bel W3, while in the cv. Bel B both parameters recovered in the post-fumigation period. Thus, ozone tolerance may be related to a sustained capacity of recovery. There was no linear correlation between ozone uptake and photosynthesis reduction, but a threshold of ozone uptake was found after which photosynthesis was substantially impaired. This threshold may or may not be reached under the same external ozone level, indicating that the AOT40 may not be a sufficiently accurate index for the detection of ozone damage in plants.     

Species- and age-dependent sensitivity to ozone in young plants of pea,wheat and spinach: Effects on acyl lipid and pigment content and metabolism

Acyl lipids and pigments were analyzed in young plants of garden pea, spring wheat and spinach exposed to < 5 or 65 nl l^?1 ozone 12 h per day for 6 days. In one set of experiments, the plants were exposed to ¹⁴CO₂ for 2 h 3 days prior to ozone exposure. The plants responded differently to the moderately enhanced level of ozone used Spinach was not at all sensitive while in both pea and wheat, leaves of different ages differed in ozone sensitivity. In pea, ozone sensitivity increased with leaf age. In the second and third oldest leaves, the amounts of galactolipids per leaf area and the proportions of 18:3 of the total lipid extract and of phosphatidylglycerol decreased. In the second oldest leaf, ozone also caused a decreased proportion of 18:3 of monogalactosyldiacylglycerol. In the fourth oldest leaf, lipid composition and galactolipid unsaturation was unaffected, but ozone caused decreased leaf expansion resulting in increased acyl lipid content per leaf area. In both the first and second leaves of wheat, ozone fumigation caused a marked decrease in the content of monogalactosyldiacylglycerol and in the first leaf, the contents of phosphatidylcholine and phosphatidylethanolamine increased. The proportion of 18:3 in phosphatidylcholine was larger in ozone-fumigated than in control plants, while the reverse applied for phosphatidylglycerol. In the oldest sampled leaves of pea and wheat, ozone caused an increase in the radioactivity associated with β-carotene, indicating increased turnover. Thus, while spinach was unaffected, in both pea and wheat ozone caused a decrease in the proportion of chloroplast membrane lipids to non-chloroplast membrane lipids in older leaves while younger leaves were less sensitive.     

Responses of transgenic poplar (Populus tremula x P. alba) overexpressing glutathione synthetase or glutathione reductase to acute ozone stress: visible injury and leaf gas exchange

Untransformed hybrid poplar (Populus tremula x P. alba) and transgenic lines overexpressing glutathione synthetase (GshS) in the cytosol (200-300-fold) or glutathione reductase (GR) either in the cytosol 5-fold) or in the chloroplast (150-200-fold) were exposed to 0 (control), 100, 200 or 300 nl l^-1 ozone for 3 d for 7 h d^-1. Following acute ozone stress treatments, wild-type and transgenic poplar suffered from visible foliar injury consisting of dark brown necrotic lesions on the laminae. Necrotic lesions were sharply separated from photosynthetically active cells by a band of red-violet discoloured cell lines showing yellow autofluorescence by blue light, and blue autofluorescence by UV-light excitation. When plants were exposed to 100 nl l^-1 ozone, leaf injury was in general negligible, but when 200 and 300 nl l^-1 ozone was applied, in both untransformed poplar and transgenic lines overexpressing GshS or GR up to 60% and 80%, respectively, visible injury developed on mature leaves. The mean percentage of injured leaf area amounted to 20-30% (200 nl l^-1) and 40-60% (300 nl l^-1). Irrespective of transformation, young leaves of poplar trees were only slightly affected by ozone treatments. In support of these observations, net CO2 assimilation rates of mature leaves were decreased by up to 65% (300 nl l^-1 ozone) in wild-type and transformed poplar, whereas net photosynthesis of young leaves remained unaffected even under severe stress conditions. Leaf conductance was significantly decreased by all ozone treatments, but was in the same range in young and mature leaves, and in wild-type and transformed poplar, pre- and post-exposure to ozone. It can therefore be assumed that the ozone doses effectively taken up into the leaf tissue were not dependent on leaf development and that the strength of the ozone stress exerted was similar in all types of poplar trees investigated in this study.From these data it is concluded that: (i) elevated foliar activities of glutathione synthetase or glutathione reductase alone are not sufficient to improve tolerance of hybrid poplar to acute ozone stress, and (ii) the sensitivity of poplar leaves to acute ozone stress is controlled by unknown factors closely related to leaf development rather than by foliar activities of glutathione synthetase and glutathione reductase, or leaf conductance.     

Effects of Ozone Exposure on Resistance of Wheat Genotypes to Pyrenophora tritici-repentis

Three spring wheat genotypes, susceptible, moderately resistant or resistant to Pyrenophora tritici-repentis (tan spot fungus) were exposed to charcoal-filtered air and to approx. 80, 160, 240 (g m^?3 ozone for five consecutive days (7 h per day). Visible leaf injury on seedling plants (three-leaf stage) was only observed after fumigation with 160 or 240 (g m^?3 O₃. Amount of injury was four-fold and 10-fold on the susceptible genotype when compared to resistant or moderately resistant genotype at the two highest concentration of ozone, respectively. Genotypic differences to O₃ tolerance were detected at the seedling growth stage (three-leaf stage) and flowering stage but not at the stem elongation stage. A significant increase in tan spot lesion area was observed only on O₃ predisposed second top most leaves of the susceptible genotype at all the three levels of ozone. Predisposition did not enhance tan spot development in resistant and moderately resistant genotypes. In a test with 12 wheat genotypes, a highly significant positive correlation (r = 0· 986, p < 0· 0001) was observed between ozone sensitivity (percent leaf area damaged due to 240 (g m^?3 ozone exposure) and tan spot development (mm² lesion area) following inoculation with P. tritici-repentis. It indicates that wheat genotypes resistant to the tan spot fungus might be tolerant to ozone damage.     

Effects of low level O3 exposure on leaf diffusive conductance and water-use efficiency in hybrid poplar

Abstract Young, amphistomatous hybrid poplar (Populus deltoides x trichocarpa) plants were exposed daily to either background (0.025 cm³ m^-3) or elevated (0.125 cm³ m^-3) concentrations of O₃. Levels of abaxial and adaxial leaf conductance were affected interactively by pollutant treatment, leaf age, and photon fluence rate. Consequently, conductance in O₃-treated leaves was sometimes higher and sometimes lower than in comparable control leaves, depending on leaf age or level of photon fluence rate. For example, at low photon fluence rate or in the dark, conductance was greater in O₃-treated than in control plants, while at high photon fluence rate that relationship was reversed. Exposure to O₃ also reduced the water-use efficiency and range of leaf conductance of individual leaves, and altered the relationship between the conductances of the two leaf surfaces (the ratio of abaxial to adaxial leaf conductance was increased). Furthermore, O₃ treatment resulted in diminished stomatal control of water loss; excised O₃-treated leaves had higher conductances and wilted sooner than excised control leaves of identical ages. Overall, the data indicate that exposure to O₃ resulted in impaired stomatal function.     

Radish plant behaviour under short-term elevated ozone fumigation

Effects of short-term ozone (O₃) fumigation on radish (Raphanus sativus L.) plants were examined in growth chambers under controlled environment conditions. Plants were exposed to 0 μg/m³ (reference), 80 μg/m³, 160 μg/m³ and 240 μg/m³ O₃ concentrations for 7 h per day for five days. Day/night temperature was 21°C/14°C and photoperiod 16 h. Chlorophyll content was evaluated spectrophotometrically. Chromatographic analysis of saccharides was also undertaken. The results showed that elevated O₃ inhibited the growth of radish rhizocarps, net assimilation rate and biomass accumulation. O₃ induced leaf desiccation, necrosis and premature senescence, but a typical reaction of plants to O₃ stress was the rapid regeneration of new leaves. O₃ inhibited accumulation of carotenoids more than chlorophylls. The higher photosynthetic pigment content in newly generated radish leaves may be regarded as an adaptation of the photosynthetic system to O₃. Leaf saccharide metabolism and incorporation depended on O₃ concentration. Rapid regeneration of new leaves and increased content of photosynthetic pigments is the typical reaction of radish plants to O₃ stress.     

Ascorbate in the leaf apoplast is a factor mediating ozone resistance in Plantago major

The role of ascorbate in mediating ozone resistance was examined in Plantago major L. Seedlings of eleven populations which exhibited differential resistance to ozone were fumigated in controlled environment chambers with charcoal/Purafil®-filtered air (CFA) or CFA plus 15 nmol·mol^–1 ozone overnight rising to a maximum between 12:00–16:00 hours of 75 nmol·mol^–1 for 14 d. Measurements of ascorbate content were made on apoplastic and symplastic extracts. Populations differed in their constitutive level of ascorbate in youngest fully expanded leaves, and regression analysis revealed a significant correlation between ascorbate content in ozone-treated leaves and the ozone resistance of the populations. The relationship was stronger using apoplastic ascorbate levels than with corresponding symplastic measurements. The ascorbate content of the youngest fully expanded leaf of an ozone sensitive population was increased by foliar application of ascorbate. No significant difference in stomatal conductance was found between control and ascorbate-treated plants. Following spraying, plants were fumigated with 400 nmol·mol^–1 ozone for 7 h. In control plants, ozone exposure resulted in extensive visible leaf damage (20–70 % at the end of the fumigation period) and decreased rates of CO₂ assimilation (–57 %). However, ascorbate treatment prevented the appearance of visible injury, and ameliorated the decline in photosynthesis induced by ozone (–26 %). Modelled data estimating the extent of protection afforded by apoplastic ascorbate against ozone supported the experimental observations. The results suggested that although apoplastic ascorbate plays an important role, other factors must also contribute to the mediation of ozone resistance in P. major.     

Differentiation and structural decline in the leaves and bark of birch (Betula pendula) under low ozone concentrations

Summary Leaf and bark structure of a birch clone (Betula pendula Roth) continuously exposed to charcoal-filtered air or charcoal-filtered air plus ozone (0.05, 0.075, 0.1 l 1^-1) was investigated throughout one growing season. Increasing ozone dose influenced leaf differentiation by reducing leaf area and increasing inner leaf air space, density of cells developing into stomata, scales and hairs. When approximately the same ozone dose had been reached, macroscopical and microscopical symptoms appeared irrespective of the ozone concentration used during treatment. Structural decline began in mesophyll cells around stomatal cavities (droplet-like exudates on the cell walls), continued with disintegration of the cytoplasma and ended in cell collapse. Epidermal cells showed shrinkage of the mucilaginous layer (related to water loss). Their collapse marked the final stage of leaf decline. When subsidiary cells collapsed, guard cells passively opened for a transitory period before collapsing and closing. With increasing ozone dose starch remained accumulated along the small leaf veins and in guard cells. IIK-positive grains were formed in the epidermal cells. This contrasted with the senescent leaves, where starch was entirely retranslocated. Injury symptoms in stem and petiole proceeded from the epidermis to the cambium. Reduced tissue area indicated reduced cambial activity. In plants grown in filtered air and transferred into ozone on 20 August, injury symptoms developed faster than in leaves formed in the presence of ozone. Results are discussed with regard to O₃-caused acclimation and injury mechanisms.     

Exposure of cottonwood plants to ozone alters subsequent leaf decomposition

Summary Cottonwood saplings were exposed to ozone or charcoal-filtered air in a closed chamber. After leaf abscission, decomposition of individual leaf discs was measured in containers of stream water. Exposure of plants to 200 ppb ozone for 5 h caused early leaf abscission and changes in the chemical composition of leaves at time of abscission. Early-abscised leaves from O₃-exposed plants had higher nitrogen, but decomposed more slowly than leaves from control plants. Leaves from O₃-exposed plants that abscised at the normal time had lower nitrogen content and lower specific leaf mass than control leaves, but decomposed at the same rate as leaves from control plants. The results imply that O₃ exposure can alter fundamental processes important to the functioning of detritus-based aquatic ecosystems.     

Carbohydrate and carbon metabolite accumulation responses in leaves of ozone tolerant and ozone susceptible spinach plants after acute ozone exposure

The objective of this study was to determine whether exposure of plants to ozone (O₃) increased the foliar levels of glucose, glucose sources, e.g., sucrose and starch, and glucose-6-phosphate (G6P), because in leaf cells, glucose is the precursor of the antioxidant, L-ascorbate, and glucose-6-phosphate is a source of NADPH needed to support antioxidant capacity. A further objective was to establish whether the response of increased levels of glucose, sucrose, starch and G6P in leaves could be correlated with a greater degree of plant tolerance to O₃. Four commercially available Spinacia oleracea varieties were screened for tolerance or susceptibility to detrimental effects of O₃ employing one 6.5 hour acute exposure to 25O nL O₃ L^-1 air during the light. One day after the termination of ozonation (29 d post emergence), leaves of the plants were monitored both for damage and for gas exchange characteristics. Cultivar Winter Bloomsdale (cv Winter) leaves were least damaged on a quantitative grading scale. The leaves of cv Nordic, the most susceptible, were approximately 2.5 times more damaged. Photosynthesis (Pn) rates in the ozonated mature leaves of cv Winter were 48.9% less, and in cv Nordic, 66.2% less than in comparable leaves of their non-ozonated controls. Stomatal conductance of leaves of ozonated plants was found not to be a factor in the lower Pn rates in the ozonated plants. At some time points in the light, leaves of ozonated cv Winter plants had significantly higher levels of glucose, sucrose, starch, G6P, G1P, pyruvate and malate than did leaves of ozonated cv Nordic plants. It was concluded that leaves of cv Winter displayed a higher tolerance to ozone mediated stress than those of cv Nordic, in part because they had higher levels of glucose and G6P that could be mobilized during diminished photosynthesis to generate antioxidants (e.g., ascorbate) and reductants (e.g., NADPH). Elevated levels of both pyruvate and malate in the leaves of ozonated cv Winter suggested an increased availability of respiratory substrates to support higher respiratory capacity needed for repair, growth, and maintenance.Abbreviations ADPG-PPiase ADPglucose pyrophosphorylase - ASC L-ascorbic acid - APX ascorbate peroxidase - Ce CO₂ concentration in air in the measuring cuvette during photosynthesis measurements - Ci CO₂ concentration in the leaf intercellular spaces during photosynthesis measurement - Chl chlorophyll - DHA dehydroascorbic acid - DHA reductase dehydroascorbate reductase - DHAP dihydroxyacetone phosphate - GAP glyceraldehyde-3-phosphate - Gluc glucose - GR glutathione reductase - G_sw stomatal conductance with units as mmol H₂O m^-2 s^-1 - GSSG oxidized glutathione - GSH reduced glutathione - G1P glucose-1-phosphate - G6P glucose-6-phosphate - G6P dehydrogenase glucose-6-phosphate dehydrogenase - 6PG 6-phosphogluconate - 6PG dehydrogenase 6-phosphogluconate dehydrogenase - F6P fructose-6-phosphate - FBP fructose-1,6-bisphosphate - MAL malate - MDHA reductase monodehydroascorbate reductase - PE post-emergence - PEP phosphoenolpyruvate - PGA 3-phosphoglycerate - Pi orthophosphate - PYR pyruvate - Pn net CO₂ photoas-similation in leaves - PPFD photosynthetic photon flux density with units of mol photons m^-2 s^-1 - PPRC pentose phosphate reductive cycle - RuBP ribulose-1,5-bisphosphate - rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - SLW specific leaf weight - TCA cycle tricarboxylic acid cycle - Triose-P DHAP+GAP     

The Transport of Oleanolic Acid in Calendula officinalis

The transport of oleanolic acid synthesized from ¹⁴CO₂ in the individual leaves of Calendula officinalis plants was investigated. It was proved that the rate and the direction of the transport depend on the stage of vegetation and the position of the leaf on the plant. The greatest transport rates of oleanolic acid were observed from the upper level leaves towards the inflorescence during the flowering period. The overblowing period was characterized by a slower migration of oleanolic acid from the leaf supplied with radioactive precursor. The migration was directed mainly to the root especially from the low level leaves. Using CH₃¹⁴COONa as precursor, it was established that the root is unable to synthesize oleanolic acid.     

Photosynthetic responses to elevated CO2and O3 in field-grown potato(Solanum tuberosum)

Potato plants (Solanum tuberosum L. cv. Bintje) were grown to maturity in open-top chambers under three carbon dioxide (CO₂; ambient and 24 h d⁻¹ seasonal mean concentrations of 550 and 680 μmol mol⁻¹) and two ozone levels (O₃; ambient and an 8 h d⁻¹ seasonal mean of 50 nmol mol⁻¹). Chlorophyll content, photosynthetic characteristics, and stomatal responses were determined to test the hypothesis that elevated atmospheric CO₂ may alleviate the damaging influence of O₃ by reducing uptake by the leaves. Elevated O₃ had no detectable effect on photosynthetic characteristics, leaf conductance, or chlorophyll content, but did reduce SPAD values for leaf 15, the youngest leaf examined. Elevated CO₂ also reduced SPAD values for leaf 15, but not for older leaves; destructive analysis confirmed that chlorophyll content was decreased. Leaf conductance was generally reduced by elevated CO₂, and declined with time in the youngest leaves examined, as did assimilation rate (A). A generally increased under elevated CO₂, particularly in the older leaves during the latter stages of the season, thereby increasing instantaneous transpiration efficiency. Exposure to elevated CO₂ and/or O₃ had no detectable effect on dark-adapted fluorescence, although the values decreased with time. Analysis of the relationships between assimilation rate and intercellular CO₂ concentration and photosynthetically active photon flux density showed there was initially little treatment effect on CO₂-saturated assimilation rates for leaf 15. However, the values for plants grown under 550 μmol mol⁻¹ CO₂ were subsequently greater than in the ambient and 680 μmol mol⁻¹ treatments, although the beneficial influence of the former treatment declined sharply towards the end of the season. Light-saturated assimilation was consistently greater under elevated CO₂, but decreased with time in all treatments. The values decreased sharply when leaves grown under elevated CO₂ were measured under ambient CO₂, but increased when leaves grown under ambient CO₂ were examined under elevated CO₂. The results obtained indicate that, although elevated CO₂ initially increased assimilation and growth, these beneficial effects were not necessarily sustained to maturity as a result of photosynthetic acclimation and the induction of earlier senescence.     

Growth and maintenance respiration in leaves of northern red oak seedlings and mature trees after 3 years of ozone exposure

A two-component model of growth and maintenance respiration is used to study the response of northern red oak (Quercus rubra L.) seedlings and 32-year-old trees to sub-ambient (10 μmol h; cumulative dose based on 7 h daily mean), ambient (43 μmol h), and twice-ambient (85 μmolh) ozone. The relative growth rates (RGR) of leaves sampled from seedlings and trees were similar across treatments, as were specific leaf respiration rates (SRR). Growth coefficients estimated from the SRR versus RGR relationship averaged 25-3 mol CO2 kg^?1 leaf dry mass produced for seedlings and 21-5 mol kg^?1 for trees. Maintenance coefficients ranged from 0-89 to 1-07 mol CO₂ kg^?1 leaf dry mass d^?1 for seedlings and from 0-64 to 0-84 mol kg-1 d^?1 for trees. Neither coefficient was affected by ozone. Leaves sampled throughout the growing season also showed little response of respiration to ozone. This occurred despite a 30% reduction in net photosynthesis for trees grown at twice-ambient ozone. These results suggest that growth and maintenance respiration in young northern red oak leaves are not affected by ozone and that in older leaves injury can occur without a parallel increase in so-called ‘maintenance’ respiration.     

Long-term effects of SO2 on plants,SO2 metabolism and regulation of intracellular pH

The impact of SO₂ on the ionic balance of plants and its implications for intracellular pH regulation was studied to find explanations for long-term effects of SO₂. When sulphur, taken up as SO₂ by the shoots of plants, is not assimilated in organic compounds, but stored as sulphate, an equivalent amount of H⁺ is produced. These H⁺ ions are not buffered chemically, but removed by metabolic processes.On the basis of knowledge on metabolic buffering mechanisms a conceptual model is proposed for the removal of shoot-generated H⁺ by (i) OH^- ions, produced in the leaves when sulphate and nitrate are assimilated in organic compounds and/or by (ii) OH^- ions produced by decarboxylation of organic anions (a biochemical pH stat mechanism). The form in which nitrogen is supplied largely determines the potential of the plant to neutralize H⁺ in the leaves during SO₂ uptake by the proposed mechanisms.In field experiments with N₂ fixing Vicia faba L. crops, the increase of sulphate in the shoots of SO₂-exposed plants was equivalent in charge to the decrease of organic anion content, calculated as the difference between inorganic cation content (C) and inorganic anion content (A), indicating that H⁺ ions produced in the leaves following SO₂ uptake were partly removed by OH^- from sulphate reduction and partly by decarboxylation of organic anions.The appearance of chronic SO₂ injury (leaf damage) in the field experiment at the end of the growing period is discussed in relation to the impact of SO₂ on the processes involved in regulation of intracellular pH. It is proposed that the metabolic buffering capacity of leaf cells is related to the rates of sulphate and nitrate reduction and the import rate of organic anions, rather than to the organic anion content in the vacuoles of the leaf cells.