Influence of air pollutants and nutrient deficiency on D-1 protein content and photosynthesis in young spruce trees

Long-term fumigation with different mixtures of air pollutants, such as O₃, NO₂ and SO₂, in combination with potassium deficiency in 4-year-old spruce trees shows clearly that the photosynthetic apparatus reacts very sensitively to environmental stress. Even when no change in photosynthetic oxygen evolution of the leaves was measurable, the content of D-1 protein in the reaction center of photosystem II already increased significantly at low stress conditions. With increasing concentrations of pollutants, the content of D-1 protein decreased considerably. This pattern was observed with all combinations of gases, although they differed in effectiveness. The most potent effect on the content of D-1 protein was found with a combined fumigation of SO₂ plus NO₂. Obviously, the ratio between synthesis and degradation of the D-1 protein is influenced by stress factors. However, the content of D-1 protein was not strongly correlated with photosynthetic oxygen evolution in whole needles nor with electron transport rate in isolated thylakoids. Under controlled potassium deficiency, effects similar to those observed upon ozone fumigation developed. The results suggest unspecific stress answers for the D-1 content and the photosynthetic reactions rather than specific responses.     

Effects of abiotic stresses on the turnover of the D1 reaction centre II protein

In the 1990's, evidence has accumulated that various unfavourable environmental conditions substantially affect the turnover of the D₁ protein of reaction centre II, the psb A gene product. Biochemical, molecular and physiological studies in higher plants indicate that alterations of D₁ protein turnover occur under drought, nutrition deficiency, heat, chemical stress, ozone fumigation as well as UV-B and visible photo-stresses. The behaviour of photosystem II under these various stress conditions indicates that the response of D₁ protein turnover can be interpreted as a general adaptive response to environmental extremes.     

On the response of pigments and antioxidants of Pinus halepensis seedlings to Mediterranean climatic factors and long-term ozone exposure

An experiment was carried out in open-top chambers located in eastern Spain. One-yr-old Pinus halepensis Mill. seedlings were exposed during three consecutive summers to the following ozone (O₃) treatments: charcoal-filtered air (CFA), non-filtered air (NFA) or non-filtered air plus 40 nl l⁻¹ O₃, 9 h d⁻¹, 5 d wk⁻¹ (NFA+40). Seasonal variations in Aleppo pine performance were observed since reductions in chlorophyll and cellular peroxidase levels associated with increases in superoxide dismutase activity, were recorded during the summer. Similarly, a reduction in epoxidation state was found at midday during the summer, derived from an activation of the xanthophyll cycle associated to an increment in radiation and temperature levels.
The first O₃-induced effects were recorded in previous-year needles (1991) during the first summer exposure as an increase in extracellular and total peroxidase activities and in zeaxanthin levels in the NFA+40 treatment along with a trend to a higher SOD activity in this treatment. A carry-over effect was detected since a lower winter recovery of chlorophyll levels was found in the NFA+40 seedlings along with a reduction of xanthophyll levels. A reduction in chlorophyll levels was observed in the previous-year needles (1992) from the NFA+40 treatment at the end of the second fumigation period. Realistic ozone exposures induced alterations in plant antioxidative systems and plant pigments as shown in this paper. These observations together with the reductions in stomatal conductance and net photosynthesis recorded in the same experiment, indicate that Aleppo pine is a species sensitive to ozone.     

Survival and recovery of perennial forage grasses under prolonged Mediterranean drought: II. Water status, solute accumulation, abscisic acid concentration and accumulation of dehydrin transcripts in bases of immature leaves

Effects of ozone on spring wheat ( Triticum aestivum L. cv. Satu) were studied in an open-top chamber experiment during two growing seasons (1992–1993) at Jokioinen in south-west Finland. The wheat was exposed to filtered air (CF), non-filtered air (NF), non-filtered air+35 nl l⁻¹ ozone for 8 h d⁻¹ (NF⁺) and ambient air (AA). Each treatment was replicated five times. Two wk after anthesis, after 4 wk of ozone treatment (NF⁺, 45 nl l⁻¹ 1000–1800 hours, seasonal mean) the net CO₂ uptake of wheat flag leaves was decreased by c . 40% relative to CF and NF treatments, both initial and total activity of Rubisco and the quantity of protein-bound SH groups were decreased significantly. Added ozone also significantly accelerated flag leaf senescence recorded as a decrease in chloroplast size. The effect was significant 2 wk after anthesis, and senescence was complete after 4 wk. In the CF and NF treatments senescence was complete 5 wk after anthesis. The significant effect of ozone on the chloroplasts and net CO₂ uptake 2 wk after anthesis did not affect the grain filling rate. However, since the grain filling period was shorter for ozone fumigated plants, kernels were smaller. The decrease in 1000-grain weight explained most of the yield reduction in the plants under NF⁺ treatment. The results indicate that wheat plants are well buffered against substantial decrease in source activity, and that shortened flag leaf duration is the major factor causing ozone-induced yield loss.     

The response of sensitive and tolerant clones of Populus tremuloides to dynamic ozone exposure under controlled environmental conditions

Both sensitive and tolerant clones of aspen ( Populus tremuloides ) were exposed to ozone using four different exposure regimes under controlled environmental conditions. Based on data on ambient ozone from 10 cities in the USA, three treatments of 4-wk exposure to the same SUM06 (an accumulation of hourly O₃ concentrations greater than 0.06 ml l⁻¹) were constructed. The regimes allowed us to investigate: (a) the importance of long (3 wk, treatment 1) versus short (1 wk, treatment 2) duration of regimes with high peaks; (b) the effect of treatments with variable peak occurrence (treatments 1 and 2) versus uniform peak occurrence (treatment 3) during the exposure period. Nonfumigated control plants were maintained at ozone concentrations <10 nl l⁻¹. Bifacial black necrosis, a typical symptom of ozone injury on aspen leaves, occurred on both clones after 2 wk exposure. Up to 60% of the leaves on the sensitive clone were injured, with an average of 6% of total leaf area injured. In the tolerant clone only 10% of the leaves were injured, with less than 1% of the total leaf area symptomatic. The severity of injury was consistently greatest in treatment 2, followed by treatments 1 and 3, respectively. The interval between peak exposures was less important than the occurrence of peaks versus a stable maximum concentration. Premature leaf abscission occurred in the sensitive clone. Measures of gas exchange demonstrated reduced photosynthesis under ozone fumigation, but exposure regime was not a significant factor. Concentrations of two antioxidants, ascorbic acid and glutathione, were almost always greater in the resistant than in the sensitive clone, but the differences were not statistically significant. The levels of these antioxidants in aspen leaves did not change with ozone fumigation or leaf age.     

Net CO2 exchange of Pinus taeda shoots exposed to variable ozone levels and rain chemistries in field and laboratory settings

Net CO₂ exchange rates (CERs) were measured in seedlings of two loblotly pine ( Pinus taeda L.) families following 6- or 13-week exposures to ozone (charcoalfiltered or ambient air + O₃) and acid rain treatments (pH 3.3, 4.5 and 5.2). Ozone exposures (14 or 170 nl l⁻¹) were made in open-top chambers, and in continously stirred tank reactors (14, 160 or 320 nl l⁻¹) located in the field and laboratory, respectively. The CERs of whole shoots were measured in an open infrared gas analysis system at 6 levels of photosynthetic photon flux density (0, 33, 60, 410, 800 and 1660 μmol m⁻² s⁻¹). Treatment effects were not consistent between field- and laboratory-exposed seedlings. Ozone-treated field seedlings exhibited statistically significant reductions in light-saturated CER of 12.5 and 25% when measured at 6 and 13 weeks, respectively. Laboratory seedlings exhibited mixed responses to O₃, with one family showing reduced CER only after 6 weeks of O₃ exposure and the other only after 13 weeks (O₃ >160 nl l⁻¹ for both). After 13 weeks of exposure, pH 3.3, and 4.5 rain treatments enhanced light-saturated CER by an average of 52% over that observed in seedlings exposed to the pH 5.2 treatment. Enhanced CERs due to acid rain were of the same magnitude (3–5 μmol CO₂g⁻¹ s⁻¹) as ozone-induced CER reductions. No differences in dark respiration were detected between treatments. Although ozone and acid rain treatments altered seedling CER, the differences were not translated into altered final plant dry weights over the 13-week exposure period.     

Interactive effects of ozone and low UV‐B radiation on antioxidants in spruce (Picea abies) and pine (Pinus sylvestris) needles

To study the role of low UV‐B radiation in modulating the response of antioxidants to ozone, 4‐year‐old pine ( Pinus sylvestris L.) and spruce ( Picea abies L.) seedlings potted in natural soil, were exposed in phytochambers to fluctuating ozone concentrations between 9 and 113 nl 1⁻¹ according to field data recorded at Mt Wank (1175 m above sea level, Bavaria, Germany) and two‐times ambient O₃ levels. UV‐B radiation was either added at a biologically effective level of ca 1.2 kJ m⁻² day⁻¹ , which is close to that found in March at Mt Wank, or was excluded by filters (<0.08 kJ m⁻² day⁻¹). After one growth phase current‐year needles were collected and analysed for antioxidative enzyme activities (superoxide dismutase, SOD, EC 1.15.1.1; catalase, CAT, EC 1.11.1.6; guaiacol peroxidase, POD, EC 1.11.1.7) and soluble antioxidants (ascorbate, glutathione). CAT, POD, ascorbate and glutathione, but not SOD, were increased in needles of both species in response to twice ambient O₃ levels. UV‐B radiation in the presence of ambient O₃ caused an increase in total SOD activity in spruce but had no effects on antioxidants in pine. Twice ambient O₃ levels together with low UV‐B radiation counteracted the O₃‐induced increases in ascorbate and CAT in pine but not in spruce. Under these conditions spruce needles showed the highest antioxidative protection and revealed no indication of lipid peroxidation. Pine needles exposed to UV‐B and elevated O₃ levels showed elevated lipid peroxidation and a 5‐fold increase in dehydroascorbate, suggesting that this species was less protected and suffered higher oxidative stress than spruce.     

Photoinhibition of photosynthesis and growth responses at different light levels in psbA gene mutants of the cyanobacterium Synechococcus

Photoinhibition of photosynthesis and growth responses at diffrent light levels (10, 120 and 250 μmol m⁻² s⁻¹) were studied in psbA gene mutants R2S2C3 ( psbAI gene present) and R2K1 ( psbAIIIpsbAIII genes present) of the cyanobacterium Synechococcus sp . PCC 7942 ( Anacystis nidulans R2). Mutant R2K1 (possessing form II of the D1 protein of photosystem II) was much more resistant to photoinhibition than the mutant R2S2C3 (possessing form I of the D1 protein). At moderate inhibitory light levels (100 to 300 μmol m⁻² s⁻¹) this was largely ascribed to an increased rsistance of the photosystem II reaction cetres possessing form II of the D1 protein. However, at higher light levels the higher resistance mutant R2K1 was assigned to a higher rate of photosystem II repair, i.e. turnover of the D1 protein. Moreover, our results support the hypothesis that photoinhibition of photosystem II and photoinhibitory induced quenching are due to separate processes. Results from growth experiments show that the R2K1 mutant has a slower growth rate than the R2S2C3 mutant but shows an increased survival under high light stress conditions. It is hypothesized that high resistance to photoinhibition, though allowing a better survival under high light, is not advantageous for optimal growth.     

Electron spin resonance evidence for the formation of free radicals in plants exposed to ozone

Electron spin resonance (ESR) spectroscopy was used to demonstrate that free radicals are formed in O₃-fumigated plant leaves prior to the formation of visible leaf injury. ESR signals with a g-value of 2.0037 to 2.0043, were observed in pea ( Pisum sativum L. cv. Feltham first) and bean ( Phaseolus vulgaris L. cv. Pinto) plants that had been fumigated for 4 h with 70–300 nl l⁻¹ of ozone after they had been treated with the spin-trap N- t -butyl-α-phenylnitrone (PBN). The size of the ESR signals increased with the concentration of ozone used but the nature of the trapped radicals could not be identified. However, further experiments using an inhibitor of ethylene biosynthesis, arninoethoxyvinyl glycine (AVG), showed that the reaction between ozone and ethylene is the cause for ozone toxicity.     

Responses to ozone in 12 provenances of European beech (Fagus sylvatica): genotypic variation and chamber effects on photosynthesis and dry-matter partitioning

Seedlings of 12 provenances of European beech ( Fagus sylvatica ) were exposed to ambient, non-filtered air (NF) or NF+50 nl l⁻¹ ozone (NF50) for 8 h d⁻¹ in open-top chambers (OTCs), from 1 June to 4 October 1995. In 1996 exposure was continued from 31 May to 1 October at four levels: charcoal-filtered air (CF), NF, NF50 and NF+100 nl l⁻¹ ozone (NF100). Provenances were grown for both seasons in outside reference plots. All treatments were replicated. Ozone did not affect gas exchange in the provenances until late in the second season. NF100 reduced photosynthesis by 18% in August 1996 compared to CF. In September, photosynthesis was reduced by 22% in NF50 and by 29% in NF100. After two seasons, ozone reduced the root:shoot ratio by 24% when comparing CF and NF100; this was caused by reductions in the root biomass. Ozone did not affect height growth or stem diameter, and there were no ozone×provenance interactions for any growth parameter. There was, however, a significant ozone×provenance interaction for photosynthesis, showing northwest European provenances to be more sensitive to ozone than southeast European provenances when comparing dose–response estimates. This is interpreted in terms of genetic adaptation of the photosynthetic apparatus to regional growing conditions. Seedlings in the chambers grew 45% taller, and had 28% more shoot biomass and 29% smaller root biomass, resulting in a 44% reduction of root:shoot ratios compared to seedlings outside. Increased temperature and decreased PAR inside the chambers relative to the outside were probably the main causes for the differences. The magnitude of the chamber effects in OTCs raises doubts about conclusions drawn from ozone exposures in such chambers. This and previous ozone experiments with OTCs may have reached inaccurate conclusions concerning the size of ozone responses due to chamber effects.     

Photosynthesis of two poplar clones under long‐term exposure to ozone

The photosynthetic response was studied in two clones ( Populus deltoides × maximowiczii Eridano and Populus × euramericana I‐214), known for their differential response to ozone (O₃) in terms of visible symptoms, when exposed to O₃ (60 nl l⁻¹ 5 h day⁻¹, 7 and 15 days). The photosynthetic ability was tested using gas exchange and chlorophyll fluorescence analysis. O₃ caused a decrease in the CO₂ assimilation rate at light saturation level in mature leaves of both clones. Alterations of Chl fluorescence parameters, in particular the F_v/F_m ratio and non‐photochemical quenching were also observed. The effects were similar for both clones and it could not be concluded that differential effects on electron transport capacity were responsible for the observed reduction in photosynthesis. The reduction of photosynthetic rate in Eridano was due mainly to a reduced mesophyll activity, as evidenced by the increase in intercellular CO₂ concentration and the minimal changes in stomatal conductance. In contrast, in I‐214, stomatal effects were primarily responsible, although effects on the mesophyll cannot be excluded. Data obtained indicate that the effects observed at the mesophyll level may be attributed to indirect effects caused by membrane disorders.     

The effects of acid and acid/aluminum exposure on circulating plasma cortisol levels and other blood parameters in the rainbow trout, Salmo gairdneri

Softwater (Ca²⁺=50, Na⁺= 50(μequiv. l⁻¹) acclimated rainbow trout were fitted with chronic arterial catheters to allow for repetitive blood sampling. After 48 h recovery they were then exposed to either control (pH 6.5, Al = 0μg l⁻¹), acid (pH 4.8, Al = 0μg l⁻¹) or acid plus aluminum (pH 4.8, A1 = 112 μg l⁻¹) conditions for 72 h. Parameters measured included blood glucose, lactate, haemoglobin, haematocrit and plasma Na⁺, Cl⁻, protein and cortisol.
Exposure to pH 4'8 alone caused no mortality, a moderate ionoregulatory disturbance and a transient elevation in plasma cortisol. All other parameters were not significantly different from controls. Addition of aluminum to this exposure caused 100% mortality with a mean survival time of only 27.0 h. There was a marked decrease in plasma ions, hyperglycemia, lactate accumulation, haemoconcentration, red cell swelling, and a sharp rise in plasma cortisol becoming greatly increased as the fish neared death. The mechanism of toxicity of acute acid/aluminum exposure, the role for cortisol under such conditions, and the validity of cortisol and glucose as indicators of stress in fish are discussed.     

Chronic toxicity of ammonia to juvenile gilthead seabream Sparus aurata and related histopathological effects

Effect of 20 days exposure of juvenile gilthead seabream ( Sparus aurata ) to elevated levels of ammonia on growth and survival was examined in a continuous flow system. Suppressed growth and reduced survival were observed at concentrations of 8.2 and 13 mg l⁻¹ total ammonia-N (0.5 and 0.7 mg l⁻¹ un-ionized ammonia-N, respectively) and higher. The maximum acceptable toxic concentration (MATC) for growth was between 4.8 to 8.2 mg l⁻¹ total ammonia-N (0.3 and 0.5 mg l⁻¹ un-ionized ammonia-N, respectively). Fish exposed to high ammonia levels (13 mg l⁻¹ total ammonia-N, 0.7 mg l⁻¹ un-ionized ammonia-N) displayed clear signs of liver pathology. Existing evidence suggests that S. aurata is less sensitive to ammonia than other reported marine and freshwater fish.
Under certain conditions ammonia concentration in the intensive fish ponds in Eilat may exceed the no observed effect concentration for S. aurata .     

Differential ozone sensitivity of rice cultivars as indicated by visible injury and grain yield

Surface ozone pollution may cause reductions in rice ( Oryza sativa L.) yield. Ozone sensitivity in rice cultivars is often evaluated based on visible leaf injury at an early growth stage. However, it is not clear whether reduction in grain yield is related to visible injury. Therefore, visible damage and grain yield reduction were examined in Japanese and Asian rice cultivars exposed to ozone. In experiment 1, 3-week-old rice seedlings were exposed to ozone (min.: 20 nl·l⁻¹, max.:120 nl·l⁻¹) for 12 h in open-top chambers (OTCs). Visible leaf injury was quantified according to a leaf bronzing score. In experiment 2, rice plants were exposed to ozone in OTCs throughout the cropping season until grain harvest. Daily mean ozone concentrations were maintained at 2, 23, 28, 42, and 57 nl·l⁻¹ with a regular diurnal pattern of exposure. After harvest, grain yield was determined. Based on visible injury to the uppermost fully expanded leaf, the indica cultivar 'Kasalath' was most tolerant, and the japonica cultivar 'Kirara 397' was most sensitive to ozone. However, grain yields for both 'Kasalath' and 'Kirara 397' were significantly decreased after ozone exposure. The indica cultivar 'Jothi' suffered severe injury after ozone exposure but had no reduction in grain yield. Therefore, ozone sensitivity of rice cultivars evaluated by visible injury did not coincide with that evaluated by the reductions in grain yield. These results suggest that mechanisms that induce acute leaf injury do not relate to chronic ozone toxicity that reduces yield.     

Effect of chlorination on β-D-galactosidase activity of sewage bacteria and Escherichia coli

The effect of chlorine on β- D- galactosidase activity of sewage bacteria and Escherichia coli was studied. β- D- galactosidase activity of sewage was more resistant to chlorine than faecal coliform cultivability. At low initial dosage (0·05 mg Cl₂ l⁻¹) neither cultivability (colony-forming units (cfu)), nor enzyme activity of E. coli suspensions were severely impaired. When initial chlorine concentration was increased to 0·1 mg Cl₂ l⁻¹, the cfu number decreased whereas enzyme activity remained high, i.e. the enzyme activity calculated cfu⁻¹ increased. At higher chlorine doses both cfu and enzyme activity were reduced, but non-cultivable cells retained assayable activity after chlorination. Mean values of the enzyme activity calculated cfu⁻¹ decreased when the chlorine dosage was increased from 0·1 to 0·5 mg Cl₂ l⁻¹, but were not significantly different ( P > 0·05) for dosages of 0·2–0·7 mg Cl₂ l⁻¹. After chlorination, β- D- galactosidase activity of E. coli was less reduced than cfu and direct viable count numbers, but more reduced than 5-cyano-2-3, ditolyl tetrazolium chloride and total cell counts, and the enzyme activity represented an alternative activity parameter of chlorinated samples.     

Effect of acute and chronic ammonia and nitrite exposure on oxygen consumption and growth of juvenile big bellied seahorse

Juvenile big bellied seahorse Hippocampus abdominalis were exposed acutely and chronically to elevated ammonia and nitrite {24 h exposure: 0·01, 5·0, 10·1, 14·8 and 19·9 mg l⁻¹ total ammonia-nitrogen [TA-N] and <0·001, 74·4, 99·2 and 123·6 mg l⁻¹ [NO₂-N] nitrite-nitrogen and 35 days exposure: 0·11, 0·55, 1·67 and 3·07 mg l⁻¹ TAN and <0·001, 0·92, 4·67 and 9·10 mg NO₂-N l⁻¹}. Significant ( P <0·001) increases in oxygen consumption rate and ventilation frequency occurred at 14·8, 19·9 mg l⁻¹ TA-N and 99·2, 123·6 mg l⁻¹ NO₂-N for acutely exposed fish. Oxygen consumption rate was significantly ( P <0·05) elevated at 1·67 and 3·07 mg l⁻¹ TA-N in chronically treated fish and ventilation frequency increased significantly ( P <0·05) at 0·55, 1·67, 3·07 mg l⁻¹ TA-N and 4·59, 9·10 mg l⁻¹ NO₂-N. There were no significant differences in growth between controls and ammonia exposed fish. Mortalities occurred at 14·8, 19·9 mg l⁻¹ TA-N.     

In vitro regeneration and genetic transformation of the berberine-producing plant,Thalictrum flavum ssp. glaucum

Protocols have been developed for the in vitro regeneration and Agrobacterium -mediated genetic transformation of meadow rue, Thalictrum flavum ssp. glaucum . Ten-day-old seedlings were bisected along the embryonic axis and the cotyledons were co-cultured with various Agrobacterium tumefaciens strains for 3 days. The cotyledons were cultured on a shoot induction medium (B5 salts and vitamins, 30 g l⁻¹ sucrose, 2 mg l⁻¹ kinetin, and 3 g l⁻¹ Gelrite) containing 25 mg l⁻¹ hygromycin B as the selection agent and 250 mg l⁻¹ timentin to facilitate the elimination of Agrobacterium . Only the oncogenic A. tumefaciens strains A281 and C58 produced transgenic T. flavum callus tissues. A281 was the most effective strain producing hygromycin-resistant callus on 85% of the explants. Transgenic callus was subcultured on the shoot induction medium every 2 weeks. After 12 weeks, hygromycin-resistant shoots that formed on explants exposed to strain A281 were transferred to a root induction medium (B5 salts and vitamins, 25 mg l⁻¹ hygromycin B, 250 mg l⁻¹ timentin, and 3 g l⁻¹ Gelrite). Detection of the β -glucuronidase ( GUS ) gene using a polymerase chain reaction assay, the high levels of GUS mRNA and enzyme activity, and the cytohistochemical localization of GUS activity confirmed the genetic transformation of callus cultures and regenerated plants. The transformation process did not alter the normal content of berberine in transgenic roots or cell cultures; thus, the reported protocol is valuable to study the molecular and metabolic regulation of protoberberine alkaloid biosynthesis.     

Kinetic resolution of different recovery phases of photoinhibited photosystem II in cold-acclimated and non-acclimated spinach leaves

Leaf discs from spinach were exposed to a photon flux density of 1250 μmol m⁻²s⁻¹ at 5°C for 2 or 3 h in ambient air. Photoinhibition of photosystem II (PS II) was measured by means of chlorophyll fluorescence. Recovery of photosystem II was followed at 6°C and 20°C in low light or darkness for periods up to 12 h.
The experimental setup allowed kinetic resolution of different phases of recovery. The experiments revealed a temperature dependent dark recovery phase and two distinct light- and temperature dependent phases: (1) A relatively fast, light dependent recovery phase occurred in parallel with partial recovery of basic fluorescence at 6°C and 20°C. A population of PS II centers with very slow fluorescence induction kinetics, which had accumulated during photoinhibition treatment, disappeared during this phase. This fast recovery phase is proposed to represent reactivation of photoinhibited PS II, without dissassembly or incorporation of new D₁-protein. (2) A relatively slow light-dependent recovery phase took place at 20°C, but not at 6°C. In the presence of the chloroplast translation inhibitor streptomycin, part of the 2nd phase was inhibited. This phase is proposed to involve assembly of new Photosystem II centers, which is partly dependent on de novo synthesis of D₁-reaction center protein, but presumably is also using a preexisting pool of D₁-protein. Cold acclimation of the leaves resulted in a decreased sensitivity for photoinhibition of photosystem II. Recovery of photoinhibited photosystem II at 6°C of the cold-acclimated leaves was faster than in non-acclimated leaves, but this effect can be ascribed to diminished photoinhibitory damage.     

Beech trees exposed to high CO2 and to simulated summer ozone levels: Effects on photosynthesis, chloroplast components and leaf enzyme activity

Young trees of European beech ( Fagus sylvatica L.) were exposed in a phytotron to different levels of ozone and CO₂ under the climatic simulation of one vegetation period. High ozone levels were simulated similar to high ozone concentration in the field (up to 110 ppb), while CO₂ was added as 300 ppm to the present level of ca 380 ppm. Our study describes different aspects of photosynthesis from the leaf level to the reactions of selected thylakoid components at different harvest times during growth of the beech trees under the different fumigation regimes. Ozone effects appeared in the first weeks of the treatment as a stimulation of chlorophyll fluorescence (F_v/F_m), in oxygen production and in ribulose-1,5-bisphosphate carboxylase/oxygenase activity, while the summer and early autumn harvests showed strong reductions in these parameters. Only phosphoenolpyruvate carboxylase (PEPcase) activity remained higher under high ozone. The effects of high CO₂ appeared in general as a small stimulation in enzyme activity like PEPcase in spring. However, with increasing time of fumigation, reductions of all parameters were observed. Especially chlorophylls showed strong reductions under high CO₂. The combined treatment with high ozone plus high CO₂ resulted mostly in an amelioration of the negative ozone effects, although control levels were not reached.