Response to ambient ozone of two white clover (Trifolium repens L.cv. "Regal") clones, one resistant and one sensitive, grown in a Mediterranean environment

Two clones of white clover (Trifolium repens L.) differing in ozone tolerance were grown in southern Italy during 1997 and 1998 to study the effects of ambient ozone exposure on yield, leaf morphology and water use. Ambient ozone levels were high in both years with values exceeding the threshold for leaf injury reported in the literature. In both years ozone injury was observed on the sensitive clone (NC-S) but not on the resistant one (NC-R), and leaf and stolon dry matter production was significantly lower in NC-S than in NC-R. However, it cannot be excluded that other factors, such as high temperature, interacted with the effect of ozone on biomass production. The clones differed in morphological characteristics. Lower total leaf area in NC-S plants was due to a smaller number of leaves per plant, but the average area per leaf was higher in NC-S. Specific leaf weight and net assimilation rate were higher in the more productive clone (NC-R). Cumulative plant water use was higher in NC-R in each growth period because of the larger leaf area; by contrast, water use per unit leaf area was higher in NC-S, indicating higher leaf conductance to water vapour. The results suggest that ozone significantly reduces the yield of sensitive white clover plants under well-watered conditions, and that the difference in ozone tolerance between clover clones is related to differences in leaf morphology and water use.     

Integrated biomonitoring of air quality with plants and lichens: a case study on ambient ozone from central Italy

A biennial integrated survey, based on the use of vascular plants for the bioindication of the effects of tropospheric ozone together with the use of automatic analysers of ozone, as well as the mapping of lichen biodiversity was performed in the area of Castelfiorentino (Tuscany, central Italy). Photochemically produced ozone proved to be a fundamental presence during the warm season, with maximum hourly means reaching 114 ppb, exceeding the information threshold as fixed by EU: the use of supersensitive tobacco Bel-W3 confirmed the opportunity of carrying out detailed cost-effective monitoring surveys. The potential for didactical and educational implications of this methodology are appealing. Critical levels set up for the protection of vegetation have exceeded considerably. The comparison of biomass productivity in sensitive and resistant individuals (NC-S and NC-R white clover clones, in the framework of an European network) provided evidence that ambient ozone levels are associated with relevant reduction (up to 30%) in the performance of sensitive material; effects on flowering were also pronounced. The economic assessment of such an impact deserves attention. Mapping of epiphytic lichen biodiversity – which has been used to monitor air quality worldwide – was not related to ozone geographical distribution as depicted by tobacco response.     

Suitability of Nicotiana tabacum 'Bel W3' for biomonitoring ozone in São Paulo, Brazil

Nicotiana tabacum 'Bel W3' is a widely used sensitive bioindicator for ambient ozone, but it is rarely used in tropical countries. Our goal was to determine the suitability of this plant for biomonitoring ozone in the city of S?o Paulo by evaluating the relationships between leaf necroses and ozone under field conditions and measurements of chlorophyll a fluorescence and antioxidants in plants exposed to different concentrations of ozone in closed chambers. While a weak linear relationship between leaf injury and ozone concentrations (R(2)=0.10) was determined in the field, a strong linear relationship was observed in the chamber experiments. Maximum leaf injury was observed in plants submitted to 40 ppb, which coincided with a significant decrease in fluorescence and total ascorbic acid. The relationship between leaf damage observed in the field and ozone was improved when the concentrations were limited to 40 ppb (R(2)=0.28).     

Ozone risk assessment for plants: central role of metabolism-dependent changes in reducing power

The combination of stomatal-dependent ozone flux and total ascorbate level is currently presented as a correct indicator for determining the degree of sensitivity of plants to ozone. However, the large changes in carbon metabolism could play a central role in the strategy of the foliar cells in response to chronic ozone exposure, participating in the supply of reducing power and carbon skeletons for repair and detoxification, and modifying the stomatal mode of functioning. To reinforce the accuracy of the definition of the threshold for ozone risk assessment, it is proposed to also consider the redox pool (NAD(P)H), the ratio between carboxylases and the water use efficiency as indicators of the differential ozone tolerance of plants.     

Ambient ozone effects on gas exchange and total non-structural carbohydrate levels in cutleaf coneflower (Rudbeckia laciniata L.) growing in Great Smoky Mountains National Park

Ozone-sensitive and -tolerant individuals of cutleaf coneflower (Rudbeckia laciniata L.) were compared for their gas exchange characteristics and total non-structural carbohydrates at Purchase Knob, a high elevation site in Great Smoky Mountains National Park, USA. Photosynthesis and stomatal conductance decreased with increased foliar stipple. Sensitive plants had lower photosynthetic rates for all leaves, except the very youngest and oldest when compared to tolerant plants. Stomatal conductance decreased with increasing leaf age, but no ozone-sensitivity differences were found. Lower leaves had less starch than upper ones, while leaves on sensitive plants had less than those on tolerant plants. These results show that ambient levels of ozone in Great Smoky Mountains National Park can adversely affect gas exchange, water use efficiency and leaf starch content in sensitive coneflower plants. Persistence of sensitive genotypes in the Park may be due to physiological recovery in low ozone years.     

Critical levels for ozone effects on vegetation in Europe

The evidence of detrimental effects of ozone on vegetation in Europe, and the need to develop international control policies to reduce ozone exposures which are based on the effects of the pollutant, has led to attempts to define so-called critical levels of ozone above which adverse effects on trees, crops and natural vegetation may occur. This review is a critical assessment of the scientific basis of the concepts used to define critical levels for ozone and identifies the key limitations and uncertainties involved. The review focuses on the Level I critical level approach, which provides an environmental standard or threshold to minimise the effects of ozone on sensitive receptors, but does not seek to quantify the impacts of exceeding the critical level under field conditions. The concept of using the AOT (accumulated exposure over a threshold) to define long-term ozone exposure is demonstrated to be appropriate for several economically important species. The use of 40 ppb (giving the AOT40 index) as a threshold concentration gives a good linear fit to experimental data from open-top chambers for arable crops, but it is less certain that it provides the best fit to data for trees or semi-natural communities. Major uncertainties in defining critical level values relate to the choice of response parameter and species; the absence of data for many receptors, especially those of Mediterranean areas; and extrapolation to field conditions from relatively short-term open-top chamber experiments. The derivation of critical levels for long-lived organisms, such as forest trees, may require the use of modelling techniques based on physiological data from experimental studies. The exposure-response data which have been applied to derive critical levels should not be used to estimate the impacts of ozone over large areas, because of the uncertainties associated with extrapolation from the open-top chamber method, especially for forest trees, and because of spatial variation in atmospheric and environmental conditions, which may alter ozone uptake.     

Concentrations of mercury in otters (Lutra lutra L.) in Scotland in relation to rainfall

The incidence and severity of visible foliar ozone injury on black cherry (Prunus serotina) seedlings and saplings and tall milkweed (Asclepias exaltata) plants in Great Smoky Mountains National Park (GRSM) were determined by surveys along selected trails conducted during late summer 1992. The incidence (% injured plants) of ozone injury on black cherry was 47% and the percent injured leaves/injured plant and average leaf area injured were 43 and 6%, respectively. Maximum severity (avg. leaf area of the most severely injured leaf) was 12%. Black cherry seedlings and saplings exhibiting ozone injury were taller than non-injured plants. When insect feeding was present, it occurred 96% of the time on plants with ozone injury. Significantly more injury (p=0.007) on black cherry (% injured leaves/injured black cherry) occurred in the NW section of GRSM compared with the other Park sections. Regression analyses showed no relationships in ozone injury with respect to aspect, slope or elevation. Tall milkweed was evaluated twice during August for ozone injury. The incidence (% injured plants) of ozone injury was 74 and 79% for the first and second survey, respectively. The percentage of injured leaves per plant from the first to second survey was 63 to 79%, respectively. Tall milkweeds showing ozone injury were taller than the non-injured plants. The percentage of insect-damaged plants was 50% among plants without ozone injury and 60% among ozone-injured plants. Non-injured tall milkweed had fewer flowers and/or pods than the injured plants. Mean leaf area injured increased over time, and mean maximum leaf area injured increased from 8 to 11% during the same period. Regression analyses showed no differences in ozone injury regarding aspect, slope or elevation. Our findings indicate that ozone injury is widespread throughout the Park on sensitive vegetation.     

A comparison of two different approaches for mapping potential ozone damage to vegetation. A model study

Two very different types of approaches are currently in use today for indicating risk of ozone damage to vegetation in Europe. One approach is the so-called AOTX (accumulated exposure over threshold of Xppb) index, which is based upon ozone concentrations only. The second type of approach entails an estimate of the amount of ozone entering via the stomates of vegetation, the AFstY approach (accumulated stomatal flux over threshold of Y nmol m(-2) s(-1)). The EMEP chemical transport model is used to map these different indicators of ozone damage across Europe, for two illustrative vegetation types, wheat and beech forests. The results show that exceedences of critical levels for either type of indicator are widespread, but that the indicators give very different spatial patterns across Europe. Model simulations for year 2020 scenarios suggest reductions in risks of vegetation damage whichever indicator is used, but suggest that AOT40 is much more sensitive to emission control than AFstY values.     

Variation in ozone sensitivity among clones of Betula pendula and Betula pubescens

Forty clones of Betula pendula and 6 clones of Betula pubescens, originating from southern and central Finland, were ranked in order of ozone sensitivity according to visible injuries, growth and leaf senescense under low ozone exposure. The plants were fumigated in natural climatic conditions using an open-air exposure system during two growing seasons. Control plants were grown under ambient air, and the elevated-ozone exposures were 1.6x the ambient in 1994 and 1.7x the ambient in 1995. The differences in ozone sensitivity among clones were large. Ozone tolerance was related to thicker leaves and higher stomatal density as compared to sensitive clones. Ultrastructural ozone-induced symptoms were found in chloroplasts of sensitive clones. Increased number of visibly injured leaves on fumigated plants was correlated with reduced leaf formation, foliage area, shoot dry wt and number of stomata, and increased yellowing of leaves. The results suggest that a considerable proportion of birch trees, showing high sensitivity to ozone, are at risk if ambient ozone exposures increase.     

Incidence of ozone symptoms on vegetation within a National Wildlife Refuge in New Jersey, USA

During 1993-1996 and 2001-2003, we evaluated the percentage of plants (incidence) exhibiting ozone-induced foliar symptoms on vegetation within a National Wildlife Refuge located along the Atlantic Ocean coast of New Jersey, USA. Incidence varied among plant species and years. Bioindicator plants most sensitive to ozone, across all years, included native common milkweed (Asclepias syriaca) and wild grape (Vitis spp.), as well as introduced tree-of-heaven (Ailanthus altissima). Less sensitive bioindicators included Virginia creeper (Parthenocissus quinquefolia) and winged sumac (Rhus coppolina). Black cherry (Prunus serotina) and sassafras (Sassafras albidum) were least sensitive. The greatest incidence of ozone symptoms, across all plant species, occurred in 1996, followed by 2001>1995>1994>1993>2003>2002. A model was developed that showed a statistically significant relationship between incidence of ozone symptoms and the following parameters: plant species, Palmer Drought Severity Index, and the interaction of W126 x N100 measures of ambient ozone.     

CDNA microarray assessment for ozone-stressed Arabidopsis thaliana

Various detrimental factors in the environment damage plants, resulting in growth inhibition or withering. However, it is not easy to identify causal factors by visually inspecting the damaged plants. Therefore, we have developed a sensitive and reliable method for plant diagnosis, based on measuring changes in expression of a set of genes in a DNA microarray. With this method, we have been able to detect and discriminate between plants stressed by ozone, drought, or wounding.     

Using plant biomonitors and flux modelling to develop O3 dose-response relationships in Catalonia

We used tobacco Bel-W3 biomonitoring data and ozone flux modelling (WINDEP model) with the aim of developing the absorbed dose-response relationship, and comparing this approach with the most commonly used AOT40 (the sum of hourly ozone concentrations above a cut-off of 40 ppb during daylight hours, when global radiation exceeds 50 W m(-2)) in the estimation of exposure-damage curves. Leaf damage values were more related to OAD(15 days, potential) (potential ozone absorbed dose calculated over 15 consecutive days) than to AOT40 in all the studied stations. An OAD(15 days, potential) of 180 mg m(-2) was found to be the threshold for damage to the most sensitive species in this region under well watered conditions. The results show the applicability of the flux approach for risk assessment at the local scale, the improvement of the ozone damage estimation when the potential absorbed dose is modelled and used instead of just the ozone exposure, and finally, the possibilities opened by the use of biomonitoring networks.     

Influence of ozone and ethylenediurea (EDU) on growth and yield of bean (Phaseolus vulgaris L.) in open-top field chambers

Greenhouse and ambient air experiments have shown ethylene diurea (EDU) to be a strong and specific protective suppressant of ozone injury in plants. To examine how EDU affects plant responses to various ozone (O(3)) levels under controlled field conditions, Phaseolus vulgaris L. cv. Lit was treated with 150 ppm EDU every 14 days and exposed in open-top chambers to charcoal-filtered air (CF), nonfiltered air (NF) or two cf treatments with ozone added. The ozone treatments were proportional additions of one (CF1) and two (CF2) times ambient ozone levels. The mean ozone concentrations in the CF, NF, CF1 and CF2 treatments were 0.98, 14.1, 14.98 and 31.56 nl litre(-1). A two-way split plot ANOVA revealed that shoot dry weight was significantly reduced by ozone. EDU treatment was highly significant for leaf dry weight, root dry weight and shoot dry weight, but not for pod dry weight; leading to a higher biomass of EDU-treated plants. Ozone/EDU interactions were significant for root weight only, indicating that EDU reduced growth suppression by ozone. These results show that EDU action on plant biomass could be interpreted as a delay in senescence since EDU-treated plants showed a significant decreased biomass loss even in the CF treatment.     

Evaluation of Vegetation Near Coal-burning Power Plants in Southwestern Pennsylvania. II. Ozone Injury on Foliage of Hybrid Poplar

The objective of this study was to determine if the incidence or severity of foliar injury induced by regional, ambient ozone was influenced by local emissions from a complex of coal-burning power plants in southwestern Pennsylvania. Plantings of an ozonesensitive hybrid poplar clone {Populus maximowizii x trichocarpa, clone NE 388) were established in 1972 at various distances and directions from the power plants. Foliar injury caused by ambient ozone was evaluated annually from 1973 to 1990 in early to mid- August. Data are presented for the 12-year period, 1979 to 1990 inclusive, for which the most complete data sets were available. Injury from ambient ozone varied spatially and temporally, but with little relationship to power plant location. There was an apparent negative relationship between emission trends and ozone-induced symptoms, but only for one power plant. The correlation between annual mean levels of ozone-induced stipple and frequency of days (per year) with a 1-hr ozone maximum exceeding 0.04 ppm was weak, but significant. Ozone-induced bifacial necrosis was not observed on the foliage of the hybrid poplar during the drought year of 1988 in spite of record high levels of ozone; however, ozoneinduced stipple was observed.     

Factors Affecting Ozone Sensitivity and Susceptibility Of Cotton Plants

Cotton plants were investigated to ascertain the growth period of maximum sensitivity to ozone. Plants were grown from seed in the greenhouse to obtain a two-day interval age series to about three months old. Before fumigation with 0.2 to 1.0 ppm ozone for 1 hr, plants were preconditioned under standard growth chamber conditions with day temperatures of 32°, night temperatures of 21°, relative humidity of 50%, and a light intensity of 6 X 10⁴ ergs/cm²/sec. Also, stomatal opening was monitored with a resistance hygrometer. Ozone damage to leaves was assessed by rating per cent visible damage with an arbitrary scale. Approximately 0.6-0.7 ppm ozone exposure for 1 hr was necessary for visible damage. Leaves were susceptible to ozone when about three-quarters fully expanded. Leaves appearing on older plants became progressively less sensitive. Diurnally, susceptible leaves were only sensitive after several hours in the light and then lost sensitivity toward the end of a normal day. The two most critical questions concerning the nature of ozone sensitivity and susceptibility are: (7) what is the nature of susceptibility, and (2) what is the nature of sensitivity of susceptible leaves?     

Detecting plant effects is necessary to give biological significance to ambient ozone monitoring data and predictive ozone standards

The use of mechanical monitors and passive samplers has made it possible to assess concentrations of ozone over wide areas and to develop air quality standards, like AOT40 and SUM60. Monitored ozone data and AOT40 and SUM60 are also used to predict ozone injury on local and regional scales. The data and the standards do not include or account for environmental and biological variables that affect ozone uptake and plant injury. Ground proofing via vegetation surveys must be done to verify and validate plant injury predictions. If this is not done, then the standards have no biological significance and are only exercises in air quality assessment.     

Influence of ozone pollution and climate variability on net primary productivity and carbon storage in China's grassland ecosystems from 1961 to 2000

Our simulations with the Dynamic Land Ecosystem Model (DLEM) indicate that the combined effect of ozone, climate, carbon dioxide and land use have caused China's grasslands to act as a weak carbon sink during 1961-2000. This combined effect on national grassland net primary productivity (NPP) and carbon storage was small, but changes in annual NPP and total carbon storage across China's grasslands showed substantial spatial variation, with the maximum total carbon uptake reduction of more than 400gm(-2) in some places of northeastern China. The grasslands in the central northeastern China were more sensitive and vulnerable to elevated ozone pollution than other regions. The combined effect excluding ozone could potentially lead to an increase of 14Tg C in annual NPP and 0.11Pg C in total carbon storage for the same time period. This implies that improvement in air quality could significantly increase productivity and carbon storage in China's grassland ecosystems.     

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.     

Risk assessments for forest trees: the performance of the ozone flux versus the AOT concepts

Published ozone exposure-response relationships from experimental studies with young trees performed at different sites across Europe were re-analysed in order to test the performance of ozone exposure indices based on AOTX (Accumulated exposure Over a Threshold of X nmol mol(-1)) and AF(st)Y (Accumulated Stomatal Flux above a threshold of Y nmol m(-2) s(-1)). AF(st)1.6 was superior, as compared to AOT40, for explaining biomass reductions, when ozone sensitive species with differing leaf morphology were included in the analysis, while this was not the case for less sensitive species. A re-analysis of data with young black cherry trees, subject to different irrigation regimes, indicated that leaf visible injuries were more strongly related to the estimated stomatal ozone uptake, as compared to the ozone concentration in the air. Experimental data with different clones of silver birch indicated that leaf thickness was also an important factor influencing the development of ozone induced leaf visible injury.