Effects of ozone exposure in open-top chambers on poplar (Populus nigra) and beech (Fagus sylvatica): a comparison

Rooted cuttings of poplar (Populus nigra) and seedlings of beech (Fagus sylvatica) were exposed to ozone in open-top chambers for one growing season. Three treatments were applied: charcoal-filtered (CF), non-filtered (NF) and non-filtered air plus 30 ppb (nl l(-1)) ozone (NF+). Extra ozone was only added on clear days, from 09:00 until 17:00-20:00. The AOT40s (accumulated exposure over a threshold of 40 ppb), calculated from April to September were 4055 ppb.h for the NF and 8880 ppb.h for the NF+ treatments. For poplar ozone exposure caused highly significant reductions in growth rate, light-saturated net CO(2) assimilation rate, stomatal conductance, F(v)/F(m) and chlorophyll content. The largest effects were observed in August at which time ozone concentrations were elevated. A reduction was noticed in new leaf production, while accelerated ageing and visible damage to leaves caused high leaf losses. For beech the responses were similar but less pronounced: ozone exposure resulted in non-significant growth reductions, slight changes in light-saturated photosynthesis and accelerated leaf abscission. The chlorophyll content of beech leaves was not affected by the ozone treatments. The results confirmed previous observations that fast-growing tree species, such as most poplar species and hybrids, are more sensitive and responsive to tropospheric ozone than slower-growing species, such as beech. The growth reductions observed and reported here for beech were within the range of those reported in relationship to the AOT40 (accumulated exposure over a threshold of 40 ppb) critical level for ozone.     

Effects of chronic elevated ozone exposure on gas exchange responses of adult beech trees (Fagus sylvatica) as related to the within-canopy light gradient

The effects of elevated O₃ on photosynthetic properties in adult beech trees (Fagus sylvatica) were investigated in relation to leaf mass per area as a measure of the gradually changing, within-canopy light availability. Leaves under elevated O₃ showed decreased stomatal conductance at unchanged carboxylation capacity of Rubisco, which was consistent with enhanced δ¹³C of leaf organic matter, regardless of the light environment during growth. In parallel, increased energy demand for O₃ detoxification and repair was suggested under elevated O₃ owing to enhanced dark respiration. Only in shade-grown leaves, light-limited photosynthesis was reduced under elevated O₃, this effect being accompanied by lowered F_v/F_m. These results suggest that chronic O₃ exposure primarily caused stomatal closure to adult beech trees in the field regardless of the within-canopy light gradient. However, light limitation apparently raised the O₃ sensitivity of photosynthesis and accelerated senescence in shade leaves.     

Shoot growth of mature Fagus sylvatica and Picea abies in relation to ozone

Epidemiological analysis of sequential growth data may be a tool in assessing ozone sensitivity of mature trees. Annual shoot growth of mature Fagus sylvatica in 83 Swiss permanent forest observation plots and of Picea abies in 61 plots was evaluated for 11 and 8 consecutive years, respectively, using branches harvested every 4 years. The data were assessed as annual deviation from average growth and related to fructification, ozone, meteorological parameters, and modelled soil water content using a mixed linear model. In beech, a significant association between ozone and shoot growth was observed which corresponded to a 7.4% growth reduction between 0 and 10 ppm h AOT40 (accumulated ozone over threshold 40). This is in the same order of magnitude as the response observed in experiments with seedlings. No interaction was found between ozone and drought parameters. In Norway spruce, shoot growth was neither associated with ozone nor with drought.     

Comparison of ozone uptake and sensitivity between a phytotron study with young beech and a field experiment with adult beech (Fagus sylvatica)

Chamber experiments on juvenile trees have resulted in severe injury and accelerated loss of leaves along with reduced biomass production under chronically enhanced O3 levels. In contrast, the few studies conducted on adult forest trees in the field have reported low O3 sensitivity. In the present study, young beech in phytotrons was more sensitive to O3 than adult beech in the field, although employed O3 regimes were similar. The hypotheses tested were that: (1) differences in O3 uptake were caused by the ontogenetically higher stomatal conductance of young compared to adult trees, (2) the experimental settings in the phytotrons enhanced O3 uptake compared to field conditions, and (3) a low detoxification capacity contributes to the higher O3 sensitivity of the young trees. The higher O3 sensitivity of juvenile beech in the phytotrons is demonstrated to relate to both the experimental conditions and the physiological responsiveness inherent to tree age.     

Effects of simultaneous ozone exposure and nitrogen loads on carbohydrate concentrations, biomass, growth, and nutrient concentrations of young beech trees (Fagus sylvatica)

Beech seedlings were grown under different nitrogen fertilisation regimes (0, 20, 40, and 80 kg Nha(-1)yr(-1)) for three years and were fumigated with either charcoal-filtered (F) or ambient air (O3). Nitrogen fertilisation increased leaf necroses, aphid infestations, and nutrient ratios in the leaves (N:P and N:K), as a result of decreased phosphorus and potassium concentrations. For plant growth, biomass accumulation, and starch concentrations, a positive nitrogen effect was found, but only for fertilisations of up to 40 kg Nha(-1) yr(-1). The highest nitrogen load, however, reduced leaf area, leaf water content, growth, biomass accumulation, and starch concentrations, whereas soluble carbohydrate concentrations were enhanced. The ozone fumigation resulted in reduced leaf area, leaf water content, shoot growth, root biomass accumulation, and decreased starch, phosphorus, and potassium concentrations, increasing the N:P and N:K ratios. A combined effect of the two pollutants was detected for the leaf area and the shoot elongation, where ozone fumigation amplified the nitrogen effects.     

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

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

Seasonal ozone response of mature beech trees (Fagus sylvatica) at high altitude in the Bavarian forest (Germany) in comparison with young beech grown in the field and in phytotrons

Mature beech trees (Fagus sylvatica) grown at two different altitudes in the Bavarian forest were compared with young beech trees grown at nearby field sites or in phytotrons for their macroscopic and physiological responses to different ozone (O(3)) exposures. Cumulative O(3) exposure expressed as the sum of hourly mean concentrations above the canopy ranged between 100 and 150 microl l(-1) h, with the vertical O(3) profiles at the higher altitude site being enhanced by 30%. O(3) profiles at all sites were reduced by up to 20% with increasing depth within and beneath the canopy. The leaf discoloration that developed in the absence of premature leaf loss was similar in the sun foliage of mature and young trees (including plant grown in the phytotron). Injury became apparent at low O(3) exposures, expressed as accumulated hourly means over a threshold of 40 nl l(-1) (AOT40 <3.5 microl l(-1) h) at the lower site in both the mature trees and the young beech at the field site, but only occurred when AOT40 values reached 7 microl l(-1) h at the upper site, and 6 microl l(-1) h in the phytotrons. However, the association between injury and O(3) exposure was improved when cumulative ozone uptake to sun leaves was the ozone index, used with values of about 3 mmol m(-2) resulting in visible injury in both mature and young beech growing in phytotrons. Under high ozone exposure levels of inositol were lowered, whilst concentrations of lignin-like materials were enhanced in mature beech. Similar responses were observed in young beech grown in phytotrons. As the sun foliage was affected by only a small and variable extent each year, the seasonal O(3) impact at high altitude did not appear to pose an acute risk to mature beech trees.     

Screening of bean cultivars for their response to ozone as evaluated by visible symptoms and leaf chlorophyll fluorescence

Fourteen Italian cultivars of Phaseolus vulgaris were exposed to a single pulse of ozone (O(3), 150 nl l(-1)) or to filtered air (<3 nl l(-1)) for 3.5 h. O(3) sensitivity was assessed by recording the extent of visible symptoms, effects on chlorophyll (Chl) content and changes in Chl a fluorescence parameters. This paper reports the results of an initial screening of 14 bean cultivars that was used to select a small number of cultivars for further work. Seven cultivars showed visible symptoms of injury in the range of 2-60 h after the end of the O(3) fumigation. O(3) significantly depressed total Chl content in most cultivars and a significant correlation was found between Chl content and visible symptoms. Most cultivars showed a significant change in the F(v)/F(m) ratio, even when there were no visual symptoms. There was no relationship between the extent of visual symptoms and quenching coefficients, indicating that these parameters were of no use in the determination of sensitivity to O(3) stress.     

Ozone sensitivity of Fagus sylvatica and Fraxinus excelsior young trees in relation to leaf structure and foliar ozone uptake

During the summer of 2001, 2-year-old Fraxinus excelsior and Fagus sylvatica plants were subjected to ozone-rich environmental conditions at the Regional Forest Nursery at Curno (Northern Italy). Atmospheric ozone concentrations and stomatal conductance were measured, in order to calculate the foliar fluxes by means of a one-dimensional model. The foliar structure of both species was examined (thickness of the lamina and of the individual tissues, leaf mass per area, leaf density) and chlorophyll a fluorescence was determined as a response parameter. Stomatal conductance was always greater in Fraxinus excelsior, as was ozone uptake, although the highest absorption peaks did not match the peaks of ozone concentration in the atmosphere. The foliar structure can help explain this phenomenon: Fraxinus excelsior has a thicker mesophyll than Fagus sylvatica (indicating a greater photosynthesis potential) and a reduced foliar density. This last parameter, related to the apoplastic fraction, suggests a greater ability to disseminate the gases within the leaf as well as a greater potential detoxifying capacity. As foliar symptoms spread, the parameters relating to chlorophyll a fluorescence also change. PI (Performance Index, Strasser, A., Srivastava, A., Tsimilli-Michael, M., 2000. The fluorescence transient as a tool to characterize and screen photosynthetic samples. In: Yunus, M., Pathre, U., Mohanty, P., (Eds.) Probing Photosynthesis: Mechanisms, Regulation and Adaptation. Taylor & Francis, London, UK, pp. 445-483.) has proved to be a more suitable index than Fv/Fm (Quantum Yield Efficiency) to record the onset of stress conditions.     

Growth-stage dependent crop yield response to ozone exposure

Data from four crop yield-loss field trials were examined to determine if analysis using an imposed phenological weighting function based on seasonal growth stage would provide a more accurate indication of impact of ozone exposure. Alfalfa (Medicago sativa L. cv. Moapa 69), dry bean (Phaseolus vulgaris L. cv. California Dark Red kidney), fresh market and processing tomato (Lycopersicon esculentum Mill. cv. 6718 VF and VF-145-B7879, respectively) were grown at 9-11 ambient field plots within southern California comprising an ambient gradient of ozone. The growing season for each crop was artificially divided into 'quarters' composed of equal numbers of whole days and roughly corresponding to specific growth stages. Ozone exposure was calculated for each of these 'quarters' and regressed against final crop yield using 163 different exposure statistics. Weighting functions were developed using reciprocal residual mean square (1/RMS) or percentage of the best 100 exposure statistics of the 163 tested (TOP100) for each of the quarters. The third quarter of the alfalfa season was clearly most responsive to ozone as measured by both of the weighting functions. Third quarter ozone was also weighted highest by both weighting functions for dry bean. Fresh market and processing tomato were each influenced the greatest by second quartero zone as demonstrated by both weighting functions. The occurrence of ozone during physiologically important events (flowering and initial fruit set in second quarter for tomato; pod development in third quarter for dry bean) appeared to influence the yield of these crops the greatest. Growth-stage-dependent phenological weighting of pollutant exposure may result in more effective predictions of levels of ozone exposure resulting in yield reductions.     

Visible leaf injury in young trees of Fagus sylvatica L. and Quercus robur L. in relation to ozone uptake and ozone exposure. An Open-Top Chambers experiment in South Alpine environmental conditions

An Open-Top Chambers experiment on Fagus sylvatica and Quercus robur seedlings was conducted in order to compare the performance of an exposure-based (AOT40) and a flux-based approaches in predicting the appearance of ozone visible injuries on leaves. Three different ozone treatments (charcoal-filtered; non-filtered; and open plots) and two soil moisture treatments (watered and non-watered plots) were performed. A Jarvisian stomatal conductance model was drawn up and parameterised for both species and typical South Alpine environmental conditions, thus allowing the calculation of ozone stomatal fluxes for every treatment. A critical ozone flux level for the onset of leaf visible injury in beech was clearly identified between 32.6 and 33.6 mmolO3 m(-2). In contrast, it was not possible to identify an exposure critical level using the AOT40 index. Water stress delayed the onset of the leaf visible injuries, but the flux-based approach was able to take it into account accurately.     

Quantifying ozone uptake and its effects on the stand level of common beech (Fagus sylvatica L.) in Southern Germany

Stand level O(3) fluxes were calculated using water balance calculations for 21 Common beech (Fagus sylvatica L.) stands and O(3) data from 20 monitoring stations in Southern Germany. For this intention, the daily loss of water by evapotranspiration per stand area was set against the daily O(3) uptake. During the last 30 years, O(3) uptake ranges between 0 and 187 mmol ha(-1) d(-1) per stand area. Cumulative O(3) uptake (CUO(3)), ranging between 0.1 and 0.7 mmol m(-2) yr(-1) per stand area, shows increasing trends since 1971 with considerably greater values at high altitudes. Effects in radial growth were used to derive an initial approximate critical threshold value for O(3) impacts on the vitality and growth of mature beech stands in Southern Germany. It is concluded that this concept of O(3) flux estimation in combination with dendroecological analyses offers both a site specific and regional applicable approach to derive new critical levels for O(3).     

Growth response of bigcone Douglas fir (Pseudotsuga macrocarpa) to long-term ozone exposure in southern California

Long-term radial growth of bigcone Douglas fir (Pseudotsuga macrocarpa) was studied throughout its range in the San Bernardino Mountains of southern California, where ambient ozone has been high for approximately the past 40 years. A gradient of both ozone concentration and precipitation exists from west (high) to east (low). Growth rates of bigcone Douglas fir are considerably lower since 1950 throughout the San Bernardino Mountains, with the largest growth reductions in the western part of the range where ozone exposure is highest. Needle retention is also somewhat lower at high ozone sites. Lower annual precipitation since 1950 may have some impact on long-term growth reductions, and short-term growth reductions induced by drought are an important component of long-term growth reductions at sites with high ozone exposure. An ozone-climate stress complex may be responsible for recent reductions in the growth of bigcone Douglas fir.     

Foliar response of black cherry (Prunus serotina) clones to ambient ozone exposure in central Pennsylvania

During late summer of 1996 and 1997 we examined ozone-induced foliar injury in a plantation of 111 black cherry trees (ramets) comprising 15 clones originating from wild ortets growing in the Allegheny National Forest, Pennsylvania, and the Monongahela National Forest, West Virginia. The experimental plantation was a clonal seed orchard in Centre County, Pennsylvania, started in 1971 using ortet buds grafted onto seedling rootstocks of mixed origin. Clones differed significantly in severity of foliar injury symptoms (F=31.83, p<0.001). One clone (R-12) had significantly more foliar injury with >50% leaf area affected than other clones during both years. In contrast, clone R-14, which is from the same area in northcentral Pennsylvania as R-12, exhibited significantly less injury (LAA<6%). Although ambient O(3) concentrations were similar in both years, foliar injury was significantly greater (15.7%) in 1996 than in 1997 (9.9%). This is probably explained by lower stomatal conductance in 1997 caused by drier and hotter weather patterns in June and July of that year. Despite very different weather patterns and overall levels of injury in 1996 and 1997, mean clonal injury was significantly correlated between both years of assessment (r=0.92, p<0.001). Within tree crowns, foliage in lower and inner crown positions was significantly more injured than foliage in upper and exterior crown positions. There was no evidence of geographically based population differences in sensitivity to foliar O(3) injury. On the contrary, results demonstrate that wild genotypes of proximal geographic origin may differ greatly in sensitivity.     

Ozone exposure, defoliation of beech (Fagus sylvatica L.) and visible foliar symptoms on native plants in selected plots of South-Western Europe

The relationships between crown defoliation of beech, visible foliar symptoms on native vegetation and ozone exposure were investigated on permanent monitoring sites in South-Western Europe in the years 2000-2002. Relationships between defoliation of beech and O3 (seasonal mean, 2-week maximum, AOT40) were investigated by means of multiple regression models (11 plots, 1-3 years of data each) and a model based on temporal autocorrelation of defoliation data (14 plots, 1-3 years of data each). Different multiple regression techniques were used. The four models generated (R2=0.71-0.85, explained variance in cross-validation 61-78%) identified several significant predictors of defoliation, with AOT40 (p=0.008) and foliar content of phosphorous (p=0.0002-0.0004) being common to all models. The autocorrelation model (R2=0.55; p<0.0001) was used to calculate expected defoliation on the basis of the previous year's defoliation, and model predictions were used as an estimate of expected defoliation under constant site and environmental condition. Residuals (predicted-measured) plotted against current AOT40 shows that a possible effect of ozone occurs only at very high AOT40 (>35,000 ppbh). O3-like visible foliar symptoms were recorded on 65 species at 47% of the common monitoring sites in 2001 and 38% in 2002. No relationship was found between O3 exposure, frequency of symptomatic sites and frequency of species with symptoms (R2=0.11; p>0.05). A number of questions related to the ecological and methodological basis of the survey were identified. Inherent sampling and non-sampling errors and multicollinearity of the data suggest great caution when examining results obtained from mensurational, correlative studies.     

A statistical model for FEV1 response to arbitrary dynamic ozone exposure conditions

Lung function response to inhaled ozone at ambient air pollution levels is known to be a function of ozone concentration, exposure duration, and minute ventilation. Most data-driven exposure-response models address exposures under static condition (i.e., with a constant ozone concentration and exercise pattern). Such models are simplifications, as both ambient ozone concentrations and normal human activity patterns change with time. The purpose of this study was to develop a dynamic model of response with the advantages of a statistical model (a relatively simple structure with few parameters). A previously proposed mechanistic model for changes in specific airways resistance was adapted to describe the percent change in forced expiratory volume in one second (FEV1). This model was then reduced using the fit to three existing exposure-response data sets as criterion. The resulting model consists of a single linear differential equation together with an algebraic logistic equation. Under restricted static conditions the model reduces to a logistic model presented earlier by the authors.     

Chlorophyll a fluorescence, antioxidant enzymes and lipid peroxidation in tomato in response to ozone and benomyl.

Ozone is a widely distributed phytotoxic air pollutant and is known to reduce the yield of several important agricultural crops in Spain. However, benomyl has been found to lessen the adverse impact of ozone on plants. We studied the effects of ozone and benomyl on chlorophyll a fluorescence, antioxidant enzymes, and lipid peroxidation in tomato (Lycopersicon esculentum Mill. cv. Tiny Tim) grown in open-top chambers in the field. Our results indicate that benomyl prevented the peroxidation of membrane lipids and increased protection of PSII from ozone. There was also a significant reduction in the activity of the antioxidant enzyme superoxide dismutase in ozone-exposed plants that had not been treated with benomyl. Comparing plants treated with benomyl to untreated plants we found that, on exposure to ozone, a greater fraction of light absorption energy was cycled through the photosynthetic system in benomyl-treated plants, as shown by the higher PSII-mediated electron flow and the higher fraction of open PSII reaction centers. The values analyzed in the fluorescence parameters and lipid peroxidation were similar for plants without benomyl grown in a charcoal-filtered environment and benomyl-treated plants exposed to ozone.     

Effects of ozone impact on the gas exchange and chlorophyll fluorescence of juvenile birch stems (Betula pendula Roth.)

Effects of ozone impact on gas exchange and chlorophyll fluorescence of juvenile birch (Betula pendula) stems and leaves were investigated. Significant differences in the response of leaves and stems to ozone were found. In leaves, O3 exposure led to a significant decline in photosynthetic rates, whereas stems revealed an increased dark respiration and a concomitant increase in corticular photosynthesis. In contrast to birch leaves, corticular photosynthesis appeared to support the carbon balance of stems or even of the whole-tree under O3 stress. The differences in the ozone-response between leaves and stems were found to be related to ozone uptake rates, and thus to inherent differences in leaf and stem O3 conductance.     

Spatial variability and temporal stability of throughfall deposition under beech (Fagus sylvatica L.) in relationship to canopy structure

Although the spatial variability of throughfall (TF) in forest ecosystems can have important ecological implications, little is known about the driving factors of within-stand TF variability, particularly in deciduous forests. While the spatial variability of TF water amount and H+ deposition under a dominant beech (Fagus sylvatica L.) tree was significantly higher in the leafed period than in the leafless period, the spatial TF deposition patterns of most major ions were similar in both periods. The semiannual TF depositions of all ions other than H+ were significantly positively correlated (r=0.68-0.90, p<0.05) with canopy structure above sample locations throughout the entire year. The amounts of TF water and H+ deposition during the leafed period were negatively correlated with branch cover. We conclude that the spatial heterogeneity of ion deposition under beech was significantly affected by leaves in the growing period and by branches in non-foliated conditions.     

Responses of two birch (Betula pendula Roth) clones to different ozone profiles with similar AOT40 exposure

Saplings of two clones of European white birch (Betula pendula Roth) were exposed to three different ozone profiles resulting in same AOT40 value of 13–14 ppm h in a chamber experiment. The sensitive clone 5 and the more tolerant clone 2 were growing (1) under filtered air (=control), or (2) were exposed to 70 ppb ozone for 24 h d⁻¹ (=profile 1), (3) to 100 ppb ozone for 12 h d⁻¹ at 8:00–20:00 (=profile 2), or (4) to 200 ppb ozone for 4.5 h d⁻¹ at 9:30–14:00 (=profile 3) for 20 d. The saplings were determined for growth, visible leaf injuries, stomatal conductance, and concentrations of Rubisco, chlorophyll and carotenoids. Growth responses and induction of visible foliar injuries under different ozone profiles were variable, resulting in 4–17% lower dry mass of shoot, 16–46% reduction in stem height increment and 11–43% increase in visible injuries in clone 5, which was accompanied by higher leaf turnover rate under profile 3 indicating compensation growth. In clone 2, ozone-induced responses ranged from slight stimulation in stem height growth to 13% decrease in dry mass of shoot and 2–16% increase in visible injuries. Daytime stomatal conductance rates were lowered by 14–54% in clone 5 and 9–74% in clone 2, depending on profile. The additional power-weighted analyses revealed that high peak concentrations and exposure shape were important for induction of visible injuries in both clones and reduction in stomatal conductance in clone 5, whereas growth reductions were rather related to total cumulative exposure. The results indicate that profile of ozone exposure, night-time stomatal conductance (24 h flux), and recovery time for defence and compensations reactions should not be ignored in plant response and ozone flux modelling.