Seasonal patterns of ascorbate in the needles of Scots Pine (Pinus sylvestris L.) trees: correlation analyses with atmospheric O3 and NO2 gas mixing ratios and meteorological parameters

In the present field study the role of ascorbate in scavenging the harmful atmospheric trace gases O3 and NO2 was examined. For this purpose ascorbate contents were determined in needles of adult Scots pine trees (Pinus sylvestris L.) during three consecutive years. Ascorbate contents were correlated with ambient tropospheric O3 and NO2 concentrations and with meteorological parameters. The results showed a strong correlation of atmospheric O3 but not of atmospheric NO2 concentrations with the apoplastic content of ascorbate during the seasonal course. Ascorbate contents in needle extracts did not correlate with ambient trace gas concentrations. In the apoplastic space, but not in needle extracts ascorbate contents correlate highly significantly with global radiation. From these results it is assumed that apoplastic ascorbate in Scots pine needles is adapted to the actual atmospheric O3 concentration to mediate immediate detoxification of O3, while the atmospheric O3 concentration itself is largely dependent on light intensity.     

Apoplastic ascorbate contributes to the differential ozone sensitivity in two varieties of winter wheat under fully open-air field conditions

We studied leaf apoplastic ascorbates in relation to ozone (O₃) sensitivity in two winter wheat (Triticum aestivum L.) varieties: Yangfumai 2 (Y2) and Yangmai 16 (Y16). The plants were exposed to elevated O₃ concentration 27% higher than the ambient O₃ concentration in a fully open-air field from tillering stage until final maturity. The less sensitive variety Y16 had higher concentration of reduced ascorbate in the apoplast and leaf tissue by 33.5% and 12.0%, respectively, than those in the more sensitive variety Y2, whereas no varietal difference was detected in the decline of reduced ascorbate concentration in response to elevated O₃. No effects of O₃ or variety were detected in either oxidized ascorbate or the redox state of ascorbate in the apoplast and leaf tissue. The lower ascorbate concentrations in both apoplast and leaf tissue should have contributed to the higher O₃ sensitivity in variety Y2.     

A cumulative ozone uptake-response relationship for the growth of Norway spruce saplings

Norway spruce saplings [Picea abies (L.) Karst.] were exposed during four growing seasons to different ozone treatments in open-top chambers: charcoal filtered air (CF), non-filtered air (NF) and non-filtered air with extra ozone (NF+, 1.4xambient concentrations). The CF and NF+ ozone treatments were combined with phosphorous deficiency and drought stress treatments. The total biomass of the trees was harvested at different intervals during the experimental period. The ozone uptake to current-year needles of the Norway spruce saplings was estimated using a multiplicative stomatal conductance simulation model. There was a highly significant correlation between the reduction of total biomass and the estimated cumulative ozone uptake, which did not vary when different thresholds were applied for the rate of ozone uptake. The reduction of the total biomass was estimated to 1% per 10 mmol m(-2) cumulated ozone uptake, on a projected needle area basis.     

Ozone Injury to Pinus Strobus

In a study of the ozone-induced needle blight of eastern white pine in central New York, acute injuries naturally induced on field trees during a year of relatively low ozone concentrations (1 966) were compared with injuries induced during a year of higher concentrations (1967). Injuries were more frequent and severe and were associated with higher mean ozone concentrations in 1967 than in 1966. Characteristic symptoms were induced on foliage of pine branches exposed to controlled doses of ozone as low as 7 ± 1 pphm for four hours or 3 ± 1 pphm for 48 hours. Such doses were equalled or exceeded two and four times, respectively, in the field during the 1 967 season. Ozone sensitivity of pine needle tissue was increased by fumigation in atmospheres containing water mist. High concentrations of ozone (40-60 pphm) caused general injury of foliage of both ozone-susceptible and resistant trees; the symptoms were unlike those found in the field or caused by fumigation with low concentrations of ozone.     

Response of cellular antioxidants to ozone in wheat flag leaves at different stages of plant development

The effect of ozone (O3) on growth, yield and foliar antioxidants of spring wheat (Triticum aestivum L. cv. Turbo) was investigated in 1990 and 1991 in Braunschweig, Germany. Plants were grown full-season in pots in open-top chambers ventilated with charcoal-filtered (CF) air to which one or two levels of O3 were added. Mean 8 h day(-1) (10.00-18.00 h) O3 concentrations in the CF and CF + O3 treatments were 5.9, 61.2 and 92.5 nl litre(-1) in 1990, and 4.7 and 86.4 nl litre(-1) in 1991. Plants that received the high O3 level showed symptoms of premature senescence of the oldest leaves and yield reductions in both growing seasons. The contents of ascorbate and glutathione and the enzyme activities of ascorbate peroxidase and glutathione reductase were measured in symptomless flag leaves in weekly intervals before and after the beginning of anthesis. Leaf age had a significant effect on all antioxidants investigated. The O3 exposures of about 90 nl litre(-1) increased the activity of ascorbate peroxidase and the concentration of glutathione, but there were no pollutant effects on ascorbate content and glutathione reductase activity. Measurements of the antioxidant levels throughout one day showed no clear indications of diurnal changes in the antioxidative capacity in wheat flag leaves. The results are discussed in relation to the role of antioxidants in O3 detoxification.     

Assessment of the ozone tolerance of two soybean cultivars (Glycine max cv. Sambaíba and Tracajá) cultivated in Amazonian areas

Brazilian soybean cultivars (Glycine max Sambaíba and Tracajá) routinely grown in Amazonian areas were exposed to filtered air (FA) and filtered air enriched with ozone (40 and 80 ppb, 6 h/day for 5 days) to assess their level of tolerance to this pollutant by measuring changes in key biochemical, physiological, and morphological indicators of injury and in enzymatic and non-enzymatic antioxidants. Sambaíba plants were more sensitive to ozone than Tracajá plants, as revealed by comparing indicator injury responses and antioxidant stimulations. Sambaíba exhibited higher visible leaf injury, higher stomatal conductance, and a severe decrease in the carbon assimilation rate. Higher ozone level (80 ppb) caused an increase in cell death in both cultivars. Levels of malondialdehyde and hydrogen peroxide also increased in Tracajá exposed under 80 ppb. Sambaíba plants exhibited decreases in ascorbate and glutathione levels and in enzymatic activities associated with these antioxidants. The higher tolerance of the Tracajá soybean appeared to be indicated by reduced physiological injuries and lower stomatal conductance, which might decrease the influx of ozone and enhance oxidation-reduction reactions involving catalase, ascorbate peroxidase, ascorbate, and glutathione, most likely stimulated by higher hydrogen peroxide.     

Surface ozone in Yosemite National Park

During the summers of 2003 and 2005, surface ozone concentrations were measured with portable ozone monitors at multiple locations in and around Yosemite National Park. The goal of these measurements was to obtain a comprehensive survey of ozone within Yosemite, which will help modelers predict and interpolate ozone concentrations in remote locations and complex terrain. The data from the portable monitors were combined with concurrent and historical data from two long-term monitoring stations located within the park (Turtleback Dome and Merced River) and previous investigations with passive samplers. The results indicate that most sites in Yosemite experience roughly similar ozone concentrations during well-mixed daytime periods, but dissimilar concentrations at night. Locations that are well exposed to the free troposphere during evening hours tend to experience higher (and more variable) nocturnal ozone concentrations, resulting in smaller diurnal variations and higher overall ozone exposures. Locations that are poorly exposed to the free troposphere during nocturnal periods tend to experience very low evening ozone, yielding larger diurnal variations and smaller overall exposures. Ozone concentrations are typically highest for the western and southern portions of the park and lower for the eastern and northern regions, with substantial spatial and temporal variability. Back-trajectory analyses suggest that air with high ozone concentrations at Yosemite often originates in the San Francisco Bay Area and progresses through the Central California Valley before entering the park.     

Use of ethylenediurea (EDU) to ameliorate ozone effects on purple coneflower (Echinacea purpurea)

Purple coneflower plants (Echinacea purpurea) were placed into open-top chambers (OTCs) for 6 and 12 weeks in 2003 and 2004, respectively, and exposed to charcoal-filtered air (CF) or twice-ambient (2x) ozone (O3) in 2003, and to CF, 2x or non-filtered (NF), ambient air in 2004. Plants were treated with ethylenediurea (EDU) weekly as a foliar spray. Foliar symptoms were observed in >95% of the plants in 2x-treated OTCs in both years. Above-ground biomass was not affected by 2x treatments in 2003, but root and total-plant biomass decreased in 2004. As a result of higher concentrations of select cell wall constituents (% ADF, NDF and lignin) nutritive quality was lower for plants exposed to 2x-O3 in 2003 and 2004 (26% and 17%, respectively). Significant EDU x O3 interactions for concentrations of cell wall constituents in 2003 indicated that EDU ameliorated O3 effects on nutritive quality. Interactions observed in 2004 were inconsistent.     

Elevated atmospheric ozone increases concentration of insecticidal Bacillus thuringiensis (Bt) Cry1Ac protein in Bt Brassica napus and reduces feeding of a Bt target herbivore on the non-transgenic parent

Sustained cultivation of Bacillus thuringiensis (Bt) transgenic crops requires stable transgene expression under variable abiotic conditions. We studied the interactions of Bt toxin production and chronic ozone exposure in Bt cry1Ac-transgenic oilseed rape and found that the insect resistance trait is robust under ozone elevations. Bt Cry1Ac concentrations were higher in the leaves of Bt oilseed rape grown under elevated ozone compared to control treatment, measured either per leaf fresh weight or per total soluble protein of leaves. The mean relative growth rate of a Bt target herbivore, Plutella xylostella L. larvae was negative on Bt plants in all ozone treatments. On the non-transgenic plants, larval feeding damage was reduced under elevated ozone. Our results indicate the need for monitoring fluctuations in Bt toxin concentrations to reveal the potential of ozone exposure for altering dosing of Bt proteins to target and non-target herbivores in field environments experiencing increasing ozone pollution.     

The antioxidative response system in Glycine max (L.) Merr. exposed to Deltamethrin, a synthetic pyrethroid insecticide

Forty-five-day-old plants of Glycine max (soybean) were exposed to several Deltamethrin (synthetic pyrethroid insecticide) concentrations (0.00%, 0.05%, 0.10%, 0.15% and 0.20%) through foliar spray in the field conditions. In the treated plants, as observed at the pre-flowering (10 DAT), flowering (45 DAT) and post-flowering (70 DAT) stages, lipid peroxidation, proline content and total glutathione content increased, whereas the total ascorbate content decreased, as compared with the control. Among the enzymatic antioxidants, activity of superoxide dismutase, ascorbate peroxidase and glutathione reductase increased significantly whereas that of catalase declined markedly in relation to increasing concentration of Deltamethrin applied. The changes observed were dose-dependent, showing a strong correlation with the degree of treatment.     

Effects of simultaneous ozone exposure and nitrogen loads on carbohydrate concentrations, biomass, and growth of young spruce trees (Picea abies)

Spruce saplings were grown under different nitrogen fertilization regimes in eight chamberless fumigation systems, which were fumigated with either charcoal-filtered (F) or ambient air (O3). After the third growing season trees were harvested for biomass and non-structural carbohydrate analysis. Nitrogen had an overall positive effect on the investigated plant parameters, resulting in increased shoot elongation, biomass production, fine root soluble carbohydrate concentrations, and also slightly increased starch concentrations of stems and roots. Only needle starch concentrations and fine root sugar alcohol concentrations were decreased. Ozone fumigation resulted in needle discolorations and affected most parameters negatively, including decreased shoot elongation and decreased starch concentrations in roots, stems, and needles. In fine roots, however, soluble carbohydrate concentrations remained unaffected or increased by ozone fumigation. The only significant interaction was an antagonistic effect on root starch concentrations, where higher nitrogen levels alleviated the negative impact of ozone.     

Simulations of stomatal conductance and ozone uptake to Norway spruce saplings in open-top chambers

A simulation model was developed to estimate the stomatal conductance and ozone flux to Norway spruce saplings in open-top chambers. The model was parameterized against needle conductance measurements that were made on 4-6-year-old spruce saplings, grown in open-top chambers, in July-September during three different seasons. The spruce saplings were either maintained well watered or subject to a 7-8 week drought period in July-September each year. The simulated conductance showed a good agreement with the measured conductance for the well-watered as well as the drought stress-treated saplings. The simulations were significantly improved when different vapour pressure deficit (VPD) functions were applied for well-watered and drought-stressed spruce saplings. The cumulated ozone uptake which was calculated from the conductance simulations showed less variation between years, compared to the cumulative ozone exposure index AOT40 (accumulated exposure over a threshold of 40 ppb or nl l(-1)) for the corresponding time periods. Measurements in May 1995 demonstrated the occurrence of long-term 'memory-effects' from the drought stress treatments on the conductance. Memory-effects need to be considered when simulation models for stomatal conductance are to be applied to long-lived forest trees under a multiple stress situation.     

Estimated tropospheric ozone levels on the southeast Spanish Mediterranean coast

The objective of this study is to estimate the spatial variability of tropospheric ozone in an area using a simple model. The area in this case was applied in the western Mediterranean basin. The study period was from May to September in 2003 and 2004.A multiple linear regression between ozone concentrations, altitude and distance to the precursor sources in a fluvial basin can be used to estimate ozone values at other sites during the warmer seasons. The correlation coefficients obtained with 2-week ozone values measured at five points with a passive sampling technique were high enough to apply the model (0.77<r<0.99).To verify the model, ozone concentrations were measured with passive samplers and continuous analyzers at some selected sites, and the values were compared with the estimated concentration. The results of the validations were satisfactory, in 80% of the measurements the estimated levels differ from measured less than 20%, which is included in the bound error for the type of sampler used in this study.     

Relationship between foliar injury and changes in antioxidant levels in red and Norway spruce exposed to acidic mists

Elevated levels of total glutathione and enhanced activities of glutathione reductase and ascorbate peroxidase were found in needles of red spruce which had been exposed to acidic mists. Reduced levels of ascorbate were also detected. Such observations suggest that oxidative stress is involved in processes which resist foliar injury caused by acidic misting. Different ionic compositions of the acidic mists applied had pronounced effects on the levels of these antioxidants and the activities of related enzymes. Sulphate was found to be most effective in causing increases in glutathione, while additions of ammonium and/or nitrate mitigated the effects of sulphuric acid. Moreover, it is the composition of ions in the applied mists, rather than the levels of acidity, that determines the extent of the overall response of red spruce. By contrast, although acidic mists caused similar increases in glutathione content of Norway spruce needles, no statistically significant changes of ascorbate or related enzymic activities were found.     

Economic assessment of the negative impacts of ozone on crop yields and forest production. A case study of the estate Ostads Säteri in southwestern Sweden

Ground level ozone concentrations, in combination with the prevailing climate, at the estate Ostads S?teri in southwestern Sweden were estimated to reduce the yield of wheat and potato ranging between 5% and 10%. Occasionally, in years with the highest ozone concentrations and/or climatic conditions favoring high rates of ozone uptake to the leaves, yield loss levels above 10% may occur. Based on simple extrapolation, these ozone-induced reductions of crop yields at Ostads S?teri represent a potential total annual yield loss in Sweden in the range of 24.5 million Euro for wheat and 7.3 million Euro for potato, respectively. A simulation of forest growth at Ostad S?teri predicted that prevailing mean ozone exposure during 1993-2003 had the potential to reduce forest growth by 2.2% and the economic return of forest production by 2.6%. Using this value for extrapolation to the national level, the potential annual economic loss for Sweden due to negative impacts of ozone on forest production would be in the range of 56 million Euro (2004 prices).     

High-resolution modelling of AOT40 and stomatal ozone uptake in wheat and grassland: a comparison between 2000 and the hot summer of 2003 in Switzerland

The aim was to compare the ozone risk for agricultural crops in Switzerland during the hot and dry year 2003 with the more 'normal' situation in 2000. An improved version of the Ozone DEposition Model ODEM was used at a 2 x 2 km resolution. The distribution of the index AOT40 was compared with the accumulated stomatal ozone flux, AF(st). Averaged AOT40 at 2 m and at canopy height was much higher in 2003 than in 2000, but inter-annual differences in AF(st) for wheat and grasslands were small due to the limiting effect of low soil water contents in 2003. AOT40 suggested larger potential yield losses in wheat in 2003, while using AF(st) with a threshold of 6 nmol m(-2) s(-1) (AF(st)6) yielded similar estimates for both years. The data show that modelling of AF(st) can be used to differentiate ozone risks between regions and years at a national scale.     

How the redox state of tobacco 'Bel-W3' is modified in response to ozone and other environmental factors in a sub-tropical area?

This study intended to determine whether the redox state in plants of Nicotiana tabacum ‘Bel-W3’ fluctuates in response to the environmental factors in a sub-tropical area contaminated by ozone (São Paulo, SE - Brazil) and which environmental factors are related to this fluctuation, discussing their biomonitoring efficiency. We comparatively evaluated the indicators of redox state (ascorbic acid, glutathione, superoxide dismutase, ascorbate peroxidase, and glutathione reductase) and leaf injury in 17 field experiments performed in 2008. The redox state was explained by the combined effects of chronic levels of O₃ and meteorological variables 4-6 days prior to the plant sampling. Moderate leaf injury was observed in most cases. The redox state of tobacco decreases few days after their placement in the sub-tropical environment, causing them to become susceptible to oxidative stress imposed by chronic doses of O₃. Its bioindicator efficiency would not be diminished in such levels of atmospheric contamination.     

Modification of the biochemical pathways of plants induced by ozone: what are the varied routes to change?

When plants are observed under a low dose of ozone, some physiological and metabolic shifts occur. Barring extreme injury such as tissue damage or stomata closure, most of these disruptive changes are likely to have been initiated at the level of gene expression. The belief is oxidative products formed in ozone exposed leaves, e.g. hydrogen peroxide, are responsible for much of the biochemical adjustments. The first line of defense is a range of antioxidants, such as ascorbate and glutathione, but if this defense is overwhelmed, subsequent actions occur, similar to systemic acquired resistance or general wounding. Yet there are seemingly unrelated metabolic responses which are also triggered, such as early senescence. We discuss here the current understanding of gene control and signal transduction/control in order to increase our comprehension of how ozone alters the basic metabolism of plants and how plants counteract or cope with ozone.     

The glutathione status of mature Scots pines during the third season of UV-B radiation exposure

Impacts of UV-B radiation on the glutathione level were studied in mature Scots pine needles (Pinus sylvestris L.) during the third season of a UV-B field experiment. Studies were made on 4-week-old (July) to 14-week-old (September) current-year needles and 3-year-old needles which had their third UV-B-exposure season in progress. Depending on the season and the year (1996-98), the supplemental UV-B dose varied from 0.92 to 5.09 kJ m-2 day-1 UV-BBE compared to 0.47-2.44 kJ m-2 day-1 UV-BBE under the ambient treatment. Fully grown UV-B-treated current-year needles showed lower total glutathione concentrations after the vegetation period in September, whereas in UV-B-treated 3-year-old needles the total glutathione content was significantly lower and the proportion of oxidized glutathione (GSSG%) 56% higher in July. The significant differences in total glutathione in current-year needles in September and in active 3-year-old needles in July seem to indicate that the effect of enhanced UV-B radiation on glutathione status could be cumulative.     

Fine root dynamics of mature European beech (Fagus sylvatica L.) as influenced by elevated ozone concentrations

Fine root dynamics (diameter < 1 mm) in mature Fagus sylvatica, with the canopies exposed to ambient or twice-ambient ozone concentrations, were investigated throughout 2004. The focus was on the seasonal timing and extent of fine root dynamics (growth, mortality) in relation to the soil environment (water content, temperature). Under ambient ozone concentrations, a significant relationship was found between fine root turnover and soil environmental changes indicating accelerated fine root turnover under favourable soil conditions. In contrast, under elevated ozone, this relationship vanished as the result of an altered temporal pattern of fine root growth. Fine root survival and turnover rate did not differ significantly between the different ozone regimes, although a delay in current-year fine root shedding was found under the elevated ozone concentrations. The data indicate that increasing tropospheric ozone levels can alter the timing of fine root turnover in mature F. sylvatica but do not affect the turnover rate.