Chlorophyll, carotenoids and the activity of the xanthophyll cycle

Clone spruce trees (Picea abies L. Karst.) were exposed in the Hohenheim open-top chambers to low levels of O(3) and SO(2), singly and in combination, and to simulated precipitation of two pH treatments (Seufert et al., this volume). At the end of five years of continuous exposure, needles from the 13-year-old trees were sampled and analysed for pigments content by means of HPLC (high pressure liquid chromatography). The pigment content was determined for three needle age classes. Chlorophyll a content, measured on a dry weight basis, was similar for all needle age classes in the control chambers receiving only the simulated rain treatments at pH 5.0 or 4.0, and the chamber receiving O(3) and the rain treatment at pH 4.0. Also, no differences were noted in one-year-old needles in the chambers with SO(2) and simulated precipitation at pH 4.0 and SO(2) + O(3) and simulated precipitation at pH 4.0. Reductions of approximately 10 and 35% were measured in two-year-old needles from the chambers with SO(2) and precipitation at pH 4.0, and SO(2) + O(3) and precipitation at pH 4.0. The three-year-old needles from these chambers had 40% lower chlorophyll a content compared to the control chambers. No treatment effects were seen on the molar ratios of chlorophyll b, the carotenes, lutein, neoxanthin, and the sum of carotenoids involved in the xanthophyll cycle, violaxanthin + antheraxanthin + zeaxanthin, to chlorophyll [Formula: see text]. The xanthophyll cycle, assayed in one-year-old needles under defined light conditions (520 microE m(-2) s(-1), while light) was active in all samples. Needles from the control chambers and the chambers with SO(2) and with O(3) behaved similarly and differed from the SO(2) + O(3) treated needles by a 50% higher zeaxanthin content reached under light.     

Ammonium, nitrate and nitrite nitrogen and nitrate reductase enzyme activity in groundnut (Arachis hypogaea L.) fields after diazinon, imidacloprid and lindane treatments

Impacts of diazinon (O,O-diethyl O-2-isopropyl-6-methylpyrimidin-4-yl phosphorothioate), imidacloprid [1-(6-chloro-3-pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine] and lindane (1,2,3,4,5.6-hexachlorocyclohexane) treatments on ammonium, nitrate, and nitrite nitrogen and nitrate reductase enzyme activities were determined in groundnut (Arachis hypogaea L.) field for three consecutive years (1997 to 1999). Diazinon was applied for both seed- and soil-treatments but imidacloprid and lindane were used for seed treatments only at recommended rates. Diazinon residues persisted for 60 days in both the cases. Average half-lives (t1/2) of diazinon were found 29.3 and 34.8 days respectively in seed and soil treatments. In diazinon seed treatment, NH4(+), NO3(-), and NO2(-) nitrogen and nitrate reductase activity were not affected. Whereas, diazinon soil treatment indicated significant increase in NH4(+)-N in a 1-day sample, which continued until 90 days. Some declines in NO3(-)N were found from 15 to 60 days. Along with this decline, significant increases in NO2(-)N and nitrate reductase activity were found between 1 and 30 days. Imidacloprid and lindane persisted for 90 and 120 days with average half-lives (t1/2) of 40.9 and 53.3 days, respectively. Within 90 days, imidacloprid residues lost by 73.17% to 82.49% while such losses for lindane residues were found 78.19% to 79.86 % within 120 days. In imidacloprid seed-treated field, stimulation of NO3(-)N and the decline in NH4+NO2(-)-N and nitrate reductase enzyme activity were observed between 15 to 90 days. However, lindane seed treatment indicated significant increases in NH4(+)-N, NO2(-)-N and nitrate reductase activity and some adverse effects on NO3(-)N between 15 and 90 days.     

Nitrate reductase activity as an indicator of ponderosa pine response to atmospheric nitrogen deposition in the San Bernardino Mountains

Determinations of nitrate reductase (NR) activity in ponderosa pine (Pinus ponderosa Dougl. ex. Laws.) needles were performed during summer 1994 in two areas (consisting of six different sites) with different nitrogen (N) deposition levels in the San Bernardino Mountains, southern California. Nitrate reductase activity was used as an integrative indicator of atmospheric nitrogen deposition to pine trees (direct uptake of N species from the atmosphere and N transported from the soil). Deposition of nitrate (NO3-) to pine branches was measured in order to determine dry atmospheric inputs of the oxidized N species to tree foliage. High NR activity was detected in all of the experimental sites. Activity of the enzyme was significantly higher at the locations characterized by higher NO3- deposition to branches--slight positive correlation between branch deposited NO3- and NR activity was found. However, high variability of NR in time and between the experimental sites discredit the NR assay as a reliable indicator of N deposition for ponderosa pine in the field conditions. This could be caused by substantial interference from other abiotic and biotic factors with tropospheric ozone as probably the most important one.     

The effects of manganese precursors on Mn-based/TiO2 catalysts for catalytic reduction of NO with NH3

Manganese acetate (MnAc) and manganese nitrate (MnN) were employed as precursors for the preparation of MnAc)/TiO2, Mn (N)/TiO2, Mn(Ac)-Ce/TiO2, and Mn(N)-Ce/TiO2 by impregnation. These complexes were used as catalysts in the low-temperature selective catalytic reduction of NO with NH3. The influence of manganese precursors on catalyst characteristics, the reduction activity, and the stability of the catalysts to poisoning by H2O and SO2 were studied. Experiments showed that Mn(N) produced MnO2 with large grain sizes in Mn(N)/TiO2 catalyst. On the contrary, Mn(Ac) led to highly dispersed and amorphous Mn2O3 in Mn (Ac)/TiO2 catalyst, which had better catalytic activity and stability to SO2 at low temperatures. The doping of cerium reduced the differences in catalytic performance between the catalysts derived from different Mn precursors.     

Analyses of enzyme activities and other metabolic criteria after five years of fumigation

Enzymatic activity (peroxidase, glutamate dehydrogenase, glutamine synthetase), foliage buffering capacity, soluble protein and nitrogen content were measured in current and previous year needles from young spruce (Picea abies) and fir (Abies alba). The trees were exposed to low levels of SO(2) and/or O(3) and simulated acidic precipitation (pH 4.0) in open-top chambers from 1983 through 1988. Needle samples were taken during March 1988 at the end of the five-year fumigation period. Exposure to SO(2) substantially increased sulphur content in both needle age classes of spruce and fir, and concomitantly reduced the foliage buffering capacity index (BCI), whereas the combined fumigation with SO(2) and O(3) had no effect on BCI. Peroxidase activity was markedly higher in year-old needles compared to current-year needles. However, trees from the SO(2) and SO(2) + O(3) treatments exhibited statistically significant stimulated peroxidase activities. Similarly, changes in the activities of the nitrogen-metabolizing enzymes indicated an altered cellular function of the trees after the long-term pollution stress. Levels of activity of both glutamate dehydrogenase and glutamine synthetase were increased by exposure to SO(2), especially in spruce. Although glutamate dehydrogenase in spruce was affected by all treatments, such changes in activity were found in fir only with the SO(2) treatment. The highest activity of glutamine synthetase, however, occurred in the older needles of trees exposed to SO(2) + O(3). Total nitrogen concentration was either unaffected by the pollutant treatments or decreased in spruce compared to the controls. No statistically significant changes due to the fumigation were found in soluble protein concentrations. Results indicated that chronic exposure to air pollutants lead to alterations in metabolic processes in conifer needles, detectable either by changes in typical stress indicating values or by increases in ammonium assimilation capacity.     

Mineral content in the soil and tree foliage

Seedlings of fir (Abies alba Mill.) and spruce (Picea abies L. Karst.) were fumigated with SO(2), O(3) and SO(2) + O(3) in open-top chambers (OTCs) for almost 5 vegetation periods. As background stress, simulated rain of pH 4.0 was applied. Nutrient content of soil, soil solutions, and trees was investigated and balanced. In the upper partition of the soil high concentrations of exchangeable Ca(2+) were found in all chambers. The SO(2) and SO(2) + O(3) treatments led to increased Ca(2+), Mg(2+) and Mn(2+) concentrations in soil solution and the pool of exchangeable protons increased. This response was most evident in the SO(2) and SO(2) + O(3) chambers and less clear in the filtered pH 5.0 control chamber. In the SO(2) treatment increased Mn and S levels were found in the needles. Ca content in the needles showed a decreasing trend. O(3) alone had no consistent effect on needle nutrient content.     

Ozone-induced nitrate formation in needles and leaves of Picea abies, Fagus sylvatica and Quercus robur

Much attention has been paid to ozone as a major cause of novel forest decline in Europe. In combination with acidic mist, O(3) has been observed to increase ion leaching. Besides cations lake Mg(2+), Ca(2+), K(+), NH(4)(+), considerable amounts of nitrate were found to be leached by acidic mist from needles of Norway spruce. Controlled fumigation experiments, with 100, 300, and 600 microg O(3)m(-3) over 22 days continuously, have led to a nitrate accumulation of 94.1 +/- 14.8, 119.4 +/- 28.7 and 198.9 +/- 14.9 microg NO(3)(-1) g(-1) FW, respectively, in leaves of Quercus robur. Similar values were found in leaves of Fagus sylvatica and current and previous year needles of Picea abies. Nitrate levels of controls receiving charcoal filtered air were well below 40 microg NO(3)(-) g (-1) FW. Statistically significant elevated nitrate levels were observed after only 48 h of continuous fumigation with 600 microg O(3)m(-3), in all tree species tested, and after 144 h in the 100 microg O(3)m(-3) treatment. In another experiment, trees of Picea abies were kept in two charcoal (C) and two Purafil plus charcoal (P/C) ventilated chambers, and fumigated with O and 500 microg O(3)m(-3) in cabinets of each filter-type in order to eliminate NO(x) from chamber air. After 29 days of continuous ozone fumigation, NO(3)(-) accumulation in needles amounted to 102.0 +/- 37.7 and 137.4 +/- 40.5 microg g(-1) FW in P/C and C-filtered chambers, respectively. Nitrate contents of controls were below 30 microg NO(3)(-)g(-1) FW at the end of the experiment. No significant differences in NO(3)(-) accumulation between filter treatments were observed. Since NO(x) was reduced by more than 95% in the Purafil/charcoal versus the charcoal treatment, NO(3)(-) accumulation in needles can be attributed predominantly to the influence of ozone and not to direct NO(2) uptake of needles by the possible oxidation of NO to NO(2) in the presence of ozone.     

Effects of NO2 and SO2 on selective catalytic reduction of nitrogen oxides by ammonia

The selective catalytic reduction (SCR) characteristics of NO and NO(2) over V(2)O(5)-WO(3)-MnO(2)/TiO(2) catalyst using ammonia as a reducing agent have been determined in a fixed-bed reactor at 200-400 degrees C. The presence of NO(2) enhances the SCR activity at lower temperatures and the optimum ratio of NO(2)/NO(x) is found to be 0.5. During the SCR reactions, there are some side reactions occurred such as ammonia oxidation and N(2)O formation. At higher temperatures, the selective catalytic oxidation of ammonia and the nitrous oxide formation compete with the SCR reactions. The denitrification (DeNO(x)) conversion decreases at lower temperatures but it increases at higher temperatures with increasing SO(2) concentration. The presence of SO(2) in the feeds inhibits N(2)O formation.     

Summer-time distribution of air pollutants in Sequoia National Park, California

Concentrations of air pollutants were monitored during the May November 1999 period on a network of forested sites in Sequoia National Park, California. Measurements were conducted with: (1) active monitors for nitric oxide (NO), nitrogen dioxide (NO2) and ozone (O3); (2) honeycomb denuder/filter pack systems for nitric acid vapor (HNO3), nitrous acid vapor (HNO2), ammonia (NH3), sulfur dioxide (SO2), particulate nitrate (NO3-), ammonium (NH4+), and sulfate (SO4(2-)); and (3) passive samplers for O3, HNO3 and NO2. Elevated concentrations of O3 (seasonal means 41-71 ppb), HNO3 (seasonal means 0.4-2.9 microg/m3), NH3 (seasonal means 1.6-4.5 microg/m3), NO3 (1.1-2.0 microg/m3) and NH4+ (1.0-1.9 microg/m3) were determined. Concentrations of other pollutants were low. With increasing elevation and distance from the pollution source area of O3, NH3 and HNO3 concentrations decreased. Ammonia and NH4+ were dominant N pollutants indicating strong influence of agricultural emissions on forests and other ecosystems of the Sequoia National Park.     

An investigation into the acid content of aerosols in the ambient air at the Taj Mahal, Agra

A chemical analysis of suspended particulate matter (SPM) collected near the world famous Taj Mahal monument at Agra has been carried out. SPM samples collected on glass fibre filters were analysed for water-soluble sulphate, nitrate, chloride and ammonium ions. The data were derived from over 200 samples (each of 24 h), collected continuously during the winter periods (October through to March) of 1984-1985 and 1985-1986. The SO(4)(2-) and NO(3)(-) components are acidic in nature causing corrosion and effects on visibility, and so were studied in more detail. Mean values for SO(4)(2-) and NO(3)(-) derived from two-year data are 7.2 microg m(-3) and 8.2 microg m(-3), respectively. The SO(4)(2-)/SO(2) and NO(3)(-)/NO(2) ratiosobserved indicate faster conversion of SO(2) to SO(4)(2-) than NO(2) to NO(3)(-), the maximum levels being in January. Thus, both SO(4)(2-) and NO(3)(-) results appear to offer more promising indices of air quality than do SPM data alone.     

Effects of ozone on foliar leaching in Norway spruce (Picea abies L. Karst): confounding factors due to NOx production during ozone generation

Previous experiments with conifers fumigated with O(3), produced by air-operated electric discharge ozonators, have provided evidence that O(3) increases the leaching of NO(3)(-), NH(4)(+), K(+), Ca(2+), Mg(2+) and some other cations from needles, when the trees are treated with acid mist. This evidence has provided the foundation of the ozone-acid mist hypothesis of spruce decline. We report experiments with Norway spruce saplings fumigated with purified and unpurified O(3). The results show that the accumulation of NO(3)(-) in the needles arises from the rapid deposition of N(2)O(5) and HNO(3) formed from N(2) in the ozonator. An increase in removal of NH(4)(+), Na(+), Ca(2+), Mg(2+), Zn(2+) and Mn(2+) from the needles during soaking in H(2)SO(4), pH3, was also observed, which was related to the increase in NO(3)(-) but was independent of O(3) concentration. It is concluded that results of previous experiments cited in support of the ozone-acid mist hypothesis arose from effects which were at least partly caused by N(2)O(5) produced as a contaminant, and were incorrectly attributed to ozone. Other effects, such as growth stimulations, visible symptons, enhanced frost sensitivity, and infestation by pests or pathogens, which have been attributed to O(3) generated by electric discharge in air, should be interpreted with caution. Future experiments with ozone must eliminate this problem by either using O(2)-driven ozonators, or by purifying the output from air-driven ozonators using cold and/or water traps.     

Organism-induced accumulation of iron, zinc and arsenic in wetland soils

Four year old spruce (Picea abies (L.) Karst.) seedlings were planted in sand pots and supplied with nutrient solution. Three groups were formed, differing only in manganese nutrition (0.5 ppm, 2.5 ppm, 12.5 ppm, respectively). After three months, five individuals of each group were transferred to a dew chamber. For the next seven weeks the trees were sprayed in the evenings, the relative humidity overnight was kept high and the droplets were collected directly from the needles in the mornings. The trees were sprayed with HNO3 (pH 3.4) during the first three weeks to reduce the natural buffering capacity of the needles. After this time, the trees were sprayed with KCl (1 mM) solution, and NaHSO3 was added to the chamber resulting in SO2 concentrations usually between 50 and 150 microg m(-3). Needles and water samples were analysed. Foliar Ca seemed to be only a short-time buffer even under optimal Ca supply. A highly significant influence of managanese supply on manganese in needles and droplets was observed, as well as on sulphate, H+ and calcium concentrations in the droplets. The SO2 flux to trees treated with 12.5 ppm Mn was about twice as high as to trees treated with 0.5 ppm Mn. The conclusion is that this is due to a synergism between manganese leaching and catalysis of the SO2 oxidation by the leached Mn2+ ions. The results suggest a positive feedback between (moderate) acidification of soils and SO2 and NH3 inputs to terrestrial ecosystems.     

Phenolic compounds content in Pinus halepensis Mill. needles: a bioindicator of air pollution

Foliar phenol concentrations (total and simple phenols) were determined in Aleppo pine (Pinus halepensis Mill.) needles collected in June 2000, from 6 sites affected by various forms of atmospheric pollutants (NO, NO(2), NO(x), O(3) and SO(2)) monitored during two months. Results show an increase in total phenol content with exposure to sulphur dioxide and a reduction with exposure to nitrogen oxide pollution. p-Coumaric acid, syringic acid and 4-hydroxybenzoic acid concentrations increase with exposure to nitrogen oxide pollution, whereas gallic acid and vanillin decrease in the presence respectively of sulphur dioxide and ozone. This in situ work confirms the major interest of using total and simple phenolic compounds of P. halepensis as biological indicators of air quality.     

A comparative study of the effects of chloride, sulfate and nitrate ions on the rates of decomposition of H2O2 and organic compounds by Fe(II)/H2O2 and Fe(III)/H2O2

The effects of chloride, nitrate, perchlorate and sulfate ions on the rates of the decomposition of hydrogen peroxide and the oxidation of organic compounds by the Fenton's process have been investigated. Experiments were conducted in a batch reactor, in the dark at pH < or = 3.0 and at 25 degrees C. Data obtained from Fe(II)/H2O2 experiments with [Fe(II)]0/[H2O2]0 > or = 2 mol mol(-1), showed that the rates of reaction between Fe(II) and H2O2 followed the order SO4(2-) > ClO4(-) = NO3- = Cl-. For the Fe(III)/H2O2 process, identical rates were obtained in the presence of nitrate and perchlorate, whereas the presence of sulfate or chloride markedly decreased the rates of decomposition of H2O2 by Fe(III) and the rates of oxidation of atrazine ([atrazine]0 = 0.83 microM), 4-nitrophenol ([4-NP]0 = 1 mM) and acetic acid ([acetic acid]0 = 2 mM). These inhibitory effects have been attributed to a decrease of the rate of generation of hydroxyl radicals resulting from the formation of Fe(III) complexes and the formation of less reactive (SO4(*-)) or much less reactive (Cl2(*-)) inorganic radicals.     

Physiology of young Norway spruce

In the Hohenheim experiment young spruce (Picea abies L. Karst.) were exposed to low levels of SO(2) and/or O(3) and acid precipitation. At the end of a five-year experimental period (1983-88) the following physiological parameters were examined: water soluble thiols, ascorbic acid, glutathionereductase activity and pigment content. Exposure to SO(2), leads to an increase in thiol content, to a slight decrease of ascorbic acid and to a pronounced decrease of pigments. O(3) exposure increases the content of ascorbic acid and decreases the thiols and the glutathione-reductase activity with no change in the pigment content. The combined exposure to SO(2), and O(3) results in the most distinct deviations compared to the control chamber response. These needles show the highest increase of ascorbic acid and thiols, the dry weight is decreased as is the glutathione-reductase activity and the pigment content is reduced. Consequences of these physiological alterations for the plant's health are discussed.     

Crop growth along a gradient of ambient air pollution

An experiment, designed to elucidate the relative importance of SO2, NO2, O3, and other environmental factors in influencing the performance of four cultivars of Trifolium pratense L. and Hordeum vulgare L., was performed by growing plants in situ along a transect from central London into the surrounding countryside. A multiple regression analysis provided evidence of significant effects of SO2, NO2, and, to a lesser extent, O3, on vegetative and reproductive growth parameters, although these differed according to pollutant, cultivar, species, and the parameter concerned. The significance of these findings for the impact of ambient air pollution on the growth of crops in the more polluted rural areas of western Europe is suggested by the fact that mean SO2, NO2, and O3 concentrations in the experimental area are less than 0.020 (39.2 microg/m3), 0.025 (47.75 microg/m3), and 0.030 ppm (58.8 microg/m3), respectively. The value of the technique is discussed with respect to other studies on the effects of low levels of air pollution on crops.     

Influence of solar UV radiation on the nitrogen metabolism in needles of Scots pine (Pinus sylvestris L.)

Needles of 20-year-old Scots pine (Pinus sylvestris L.) saplings were studied in an ultraviolet (UV) exclusion field experiment (from 2000 to 2002) in northern Finland (67 °N). The chambers held filters that excluded both UV-B and UV-A, excluded UV-B only, transmitted all UV (control), or lacked filters (ambient). UV-B/UV-A exclusion decreased nitrate reductase (NR) activity of 1-year-old needles of Scots pines compared to the controls. The proportion of free amino acids varied in the range 1.08-1.94% of total proteins, and was significantly higher in needles of saplings grown under UV-B/UV-A exclusion compared to the controls or UV-B exclusion. NR activity correlated with air temperature, indicating a “chamber effect”. The study showed that both UV irradiance and increasing temperature are significant modulators of nitrogen (N) metabolism in Scots pine needles.     

Dry deposition of nitrogen and sulfur to Ponderosa and Jeffrey pine in the San Bernardino national forest in Southern California

Little is known about the concentrations, deposition rates, and effects of nitrogenous and sulfurous compounds in photochemical smog in the San Bernardino National Forest (SBNF) in southern California. Dry deposition of NO(3)(-) and NH(4)(+) to foliage of ponderosa pine (Pinus ponderosa Laws.) and Jeffrey pine (Pinus jeffreyi Grev. & Balf.) was correlated (R = 0.83-0.88) with historical average hourly O(3) concentations at 10 sites across an O(3) gradient in the SBNF. Mean deposition fluxes of NO(3)(-) to ponderosa and Jeffrey pine branches were 0.82 nmol M(-2)s(-1) at Camp Paivika (CP), a high-pollution site, and 0.19 nmol m(-2) s(-1) at Camp Osceola (CAO), a low-pollution site. Deposition fluxes of NH(4)(+) were 0.32 nmol m(-2) s(-1) at CP and 0.17 nmol m(-2) s(-1) at CAO, while mean values for SO(4)(2-) were 0.03 at CP and 0.02 nmol m(-2) s(-1) at CAO. Deposition fluxes to paper and nylon filters were higher in most cases than fluxes to pine branches at the same site. The results of this study suggest that an atmospheric concentration and deposition gradient of N and S compounds occurs along with the west-east O(3) gradient in the SBNF. Annual stand-level dry deposition rates for S and N at CP and CAO were estimated. Further studies are needed to determine if high N deposition loads in the SBNF significantly affect plant/soil nutrient relations, tree health, and the response of ponderosa pine to ozone.     

Anthropogenic impacts on natural nitrogen isotope variations in Pinus sylvestris stands in an industrially polluted area

Natural variations of the nitrogen isotopes 15N/14N (delta15N values) and the N concentrations of one-year-old needles from 7-12-year-old pine trees (Pinus sylvestris L.) were determined on 27 sites in the heavily polluted Leipzig-Halle region (former GDR). At three selected sites measurements were repeated over a period of 2 years. N concentrations and delta15N values in different needle age classes were compared at the three sites. The delta15N values of the N in the humus layer and the potential plant available N in the A(h) horizon of the local soil were determined. The 15N/14N isotope ratios (delta15N values) of one-year-old pine needles in the region of Leipzig-Halle were found to vary depending on their specific location by a factor of up to one order of magnitude (-9.6 per thousand to + 0.4 per thousand ). N concentrations in one-year-old pine needles varied between 0.71 and 1.38 mmol eq N g dw(-1). Pine stands with positive or slightly negative delta15N values and high N concentrations in one-year-old needles were concentrated around the cities of Leipzig and Halle and in the industrial areas. More negative delta15N values and lower N concentrations in one-year-old pine needles were found on sites at greater distances from the industrial agglomerations, mainly in the NE forested part. Site specific differences in the delta15N values of the N in the humus layer from three selected sites were similar to those found for the needles. No site specific differences, however, were found for the delta15N values of the water soluble nitrogen fraction from the mineral soil horizons of the same sites.     

CO2/H2O gas exchange parameters of one- and two-year-old needles of spruce and fir

Gas exchange was characterized in one- and two year-old spruce (Picea abies L. Karst.) and fir seedlings (Abies alba Mill.) which had been exposed to low levels of ozone, sulfur dioxide and simulated rain or a combination of all three variables in open top chambers from 1983 through 1988. The gas exchange measurements were carried out in March 1988 at the end of the five year experiment. The twigs examined did not exhibit any visible sign of injury, specifically no differences were apparent between trees under the treatments of simulated acidic rain at pH 5.0 and pH 4.0. The study of carbon dioxide response curves showed different effects of the pollutants on the tree species. One-Year-old spruce needles treated with O(3) and simulated acidic precipitation pH 4.0 showed noticeable reduction of net photosynthetic rate. Exposure to the combination O(3) and SO(2) at pH 4.0 resulted in a significant depression of photosynthesis in two-year-old needles Transpiration rate was not decreased to a similar extent. No changes either in photosynthesis or transpiration were found in spruce under fumigation with SO(2) alone. These results indicate that ozone is the principal cause of changes in photosynthetic performance of spruce. It alters mesophyll response rather than reducing stomatal conductance. The specific changes that occur in the mesophyll could be diagnosed as inactivation of a carbon fixing enzyme as well as damage of the electron transport system. Fir seem to be more tolerant to ozone. No changes in photosynthesis and transpiration following exposure to O(3) alone were found. However, SO(2) fumigation, alone or in combination with O(3), resulted in a marked decrease of photosynthetic performance. Particularly, carboxylation efficiency and also maximum carboxylation velocity were depressed indicating a reduction in carbon fixing enzyme activity. No differences between single and combined fumigation treatments regarding these variables were determined. However, parameters measured to determine changes in electron transport rate showed a higher depression in the presence of both pollutants. Transpiration also was reduced by SO(2).