Ozone stress response of leaf BVOC emission and photosynthesis in mountain birch (Betula pubescens spp. czerepanovii) depends on leaf age.

Oxidative stress from ozone (O3) causes plants to alter their emission of biogenic volatile organic compounds (BVOC) and their photosynthetic rate. Stress reactions from O3 on birch trees can result in prohibited plant growth and lead to increased BVOC emission rates as well as changes in their compound blend to emit more monoterpenes (MT) and sesquiterpenes (SQT). BVOCs take part in atmospheric reactions such as enhancing the production of secondary organic aerosols (SOA). As the compound blend and emission rate change with O3 stress, this can influence the atmospheric conditions by affecting the production of SOA. Studying the stress responses of plants provides important information on how these reactions might change, which is vital to making better predictions of the future climate. In this study, measurements were taken to find out how the leaves of mature mountain birch trees (Betula pubescens ssp. czerepanovii) respond to different levels of elevated O3 exposure in situ depending on leaf age. We found that leaves from both early and late summers responded with induced SQT emission after exposure to 120 ppb O3. Early leaves were, however, more sensitive to increased O3 concentrations, with enhanced emission of green leaf volatiles (GLV) and tendencies of both induced leaf senescence as well as poor recovery in the photosynthetic rate between exposures. Late leaves had more stable photosynthetic rates throughout the experiment and responded less to exposure at different O3 levels.

Accumulation of antimony and lead in leaves and needles of trees: The role of traffic emissions.

Antimony (Sb) is a toxic metalloid, which has been increasingly used in the brake lining of vehicles, and increased concentrations have been found in soils near abundant traffic. However, since very few investigations of Sb accumulation in urban vegetation have been undertaken there exists a knowledge gap. We studied the concentrations of Sb in leaves and needles of trees in the Gothenburg City area, Sweden. In addition, lead (Pb), also associated with traffic, was investigated. Sb and Pb concentrations of Quercus palustris leaves at seven sites with contrasting traffic intensity varied substantially, correlated with the traffic-related PAH (polycyclic aromatic hydrocarbon) air pollution at the sites and increased during the growing season. Sb but not Pb concentrations were significantly higher in needles of Picea abies and Pinus sylvestris near major roads compared to sites at larger distances. In Pinus nigra needles at two urban streets both Sb and Pb were higher compared to an urban nature park environment, emphasising the role of traffic emissions for these elements. A continued accumulation of Sb and Pb in three years old needles of Pinus nigra, two years old needles of Pinus sylvestris and eleven years old needles of Picea abies was observed. Our data suggest a pronounced link between traffic pollution and Sb accumulation in leaves and needles, where the particles carrying Sb seem not to be transported very far from the source. We also conclude that there exists a strong potential for Sb and Pb bioaccumulation over time in leaves and needles. Implications of these findings are that increased concentrations of toxic Sb and Pb are likely to prevail in environments with high traffic intensity and that Sb can enter the ecological food chain by accumulation in leaves and needles, which is important for the biogeochemical cycling.

Polycyclic aromatic hydrocarbon (PAH) accumulation in Quercus palustris and Pinus nigra in the urban landscape of Gothenburg, Sweden.

Trees have the potential to improve urban air quality as leaves and needles capture air pollutants from the air, but further empirical data has been requested to quantify these effects. We measured the concentration of 32 polycyclic aromatic hydrocarbons (PAHs) in leaves of pin oak (Quercus palustris) and needles of black pine (Pinus nigra) in the City of Gothenburg, Sweden, during the summer of 2018. Oak leaves were collected twice (June, September), while one-year-old (C + 1) and three-year-old (C + 3) pine needles were sampled in June to study the temporal development of leaf/needle PAH concentrations. Specific leaf area (SLA) was estimated, which permitted calculation of leaf/needle area-based PAH content that were compared with the mass-based concentration. In addition, the air concentration of PAHs and NO2 was measured using passive samplers. There was a strong correlation between air concentrations of PAH and NO2, indicating that the pollutants to a large degree originate from the same sources. In the oak leaves there was a significant decrease in low molecular mass PAHs (L-PAH, mainly gaseous) between June and September, but a significant increase in high molecular mass PAHs (H-PAH, mainly particle-bound). There was a strong correlation between L-PAH concentration in leaves and in air indicating an influence of equilibrium processes between ambient air and leaf. In the pine needles, there was a significant increase of both L-PAH and H-PAH in three-year-old needles compared to one-year-old needles. Pine was superior to oak in accumulating PAHs from the air, especially for L-PAHs when comparing area-based content. However, H-PAH concentrations were higher in oak leaves compared to pine needles on a leaf mass basis, emphasizing the importance of how concentrations are expressed. The results from this study can contribute to the development of urban planning strategies regarding the effect of vegetation on air quality.

Narrowing uncertainties in the effects of elevated CO2 on crops.

Plant responses to rising atmospheric carbon dioxide (CO2) concentrations, together with projected variations in temperature and precipitation will determine future agricultural production. Estimates of the impacts of climate change on agriculture provide essential information to design effective adaptation strategies, and develop sustainable food systems. Here, we review the current experimental evidence and crop models on the effects of elevated CO2 concentrations. Recent concerted efforts have narrowed the uncertainties in CO2-induced crop responses so that climate change impact simulations omitting CO2 can now be eliminated. To address remaining knowledge gaps and uncertainties in estimating the effects of elevated CO2 and climate change on crops, future research should expand experiments on more crop species under a wider range of growing conditions, improve the representation of responses to climate extremes in crop models, and simulate additional crop physiological processes related to nutritional quality.

Ozone exposure- and flux-yield response relationships for maize.

A stomatal ozone (O3) flux-response relationship for relative yield of maize was established by parameterizing a Jarvis stomatal conductance model. For the function (fVPD) describing the limitation of stomatal conductance by vapor pressure deficit (VPD, kPa), cumulative VPD during daylight hours was superior to hourly VPD. The latter function is proposed as a methodological improvement of this multiplicative model when stomatal conductance peaks during the morning and it is reduced later as it is the case of maize in this experiment. The model agreed relatively well with the measured stomatal conductance (R2 = 0.63). Based on the comparison of R2 values of the response functions, POD6 (Phytotoxic Ozone Dose over an hourly threshold 6 nmol m-2 s-1) and AOT40 (accumulated hourly O3 concentrations over a threshold of 40 ppb) performed similarly. The critical levels based on POD6 and AOT40 for 5% reduction in maize yield were 1.17 mmol m-2 PLA and 8.70 ppm h, respectively. In comparison with other important crops, the ranking of sensitivity of maize strongly differed depending on the O3 metric used, AOT40 or POD6. The newly proposed response functions are relevant for O3 risk assessment for this crop in Asia.

Ozone impact on wheat in Europe, Asia and North America - A comparison.

Data from experiments where field-grown wheat was exposed to ozone were collated in order to compare the effects in Europe, Asia and North America using dose-response regression. In addition to grain yield, average grain mass and harvest index were included to reflect the influence of ozone on the crop growth pattern. In order to include as many experiments as possible, daytime average ozone concentration was used as the ozone exposure index, but AOT40, estimated from average ozone concentrations, was also used to compare the performance of the two exposure metrics. The response to ozone differed significantly between the continents only for grain yield when using AOT40 as the exposure index. North American wheat was less sensitive than European and Asian that responded similarly. The variation in responses across all three continents was smallest for harvest index, followed by grain mass and grain yield. The highly consistent effect on harvest index shows that not only effects on biomass accumulation, but also on the partitioning of biomass, are important for the ozone-induced grain yield loss in wheat. The average duration of daily ozone exposure was longer in European experiments compared to North American and Asian. It cannot be excluded that this contributed to the indicated higher ozone sensitivity in European wheat in relation to North American. The main conclusions from this study are that on the average the response of wheat to ozone was lower for the older North American experiments and that the ozone response of the growth pattern reflected by grain mass and harvest index did not differ between continents.

Nitrogen application is required to realize wheat yield stimulation by elevated CO2 but will not remove the CO2 -induced reduction in grain protein concentration.

Elevated CO2 (eCO2 ) generally promotes increased grain yield (GY) and decreased grain protein concentration (GPC), but the extent to which these effects depend on the magnitude of fertilization remains unclear. We collected data on the eCO2 responses of GY, GPC and grain protein yield and their relationships with nitrogen (N) application rates across experimental data covering 11 field grown wheat (Triticum aestivum) cultivars studied in eight countries on four continents. The eCO2 -induced stimulation of GY increased with N application rates up to ~200 kg/ha. At higher N application, stimulation of GY by eCO2 stagnated or even declined. This was valid both when the yield stimulation was expressed as the total effect and using per ppm CO2 scaling. GPC was decreased by on average 7% under eCO2 and the magnitude of this effect did not depend on N application rate. The net effect of responses on GY and protein concentration was that eCO2 typically increased and decreased grain protein yield at N application rates below and above ~100 kg/ha respectively. We conclude that a negative effect on wheat GPC seems inevitable under eCO2 and that substantial N application rates may be required to sustain wheat protein yields in a world with rising CO2 .

Closing the global ozone yield gap: Quantification and cobenefits for multistress tolerance.

Increasing both crop productivity and the tolerance of crops to abiotic and biotic stresses is a major challenge for global food security in our rapidly changing climate. For the first time, we show how the spatial variation and severity of tropospheric ozone effects on yield compare with effects of other stresses on a global scale, and discuss mitigating actions against the negative effects of ozone. We show that the sensitivity to ozone declines in the order soybean > wheat > maize > rice, with genotypic variation in response being most pronounced for soybean and rice. Based on stomatal uptake, we estimate that ozone (mean of 2010-2012) reduces global yield annually by 12.4%, 7.1%, 4.4% and 6.1% for soybean, wheat, rice and maize, respectively (the "ozone yield gaps"), adding up to 227 Tg of lost yield. Our modelling shows that the highest ozone-induced production losses for soybean are in North and South America whilst for wheat they are in India and China, for rice in parts of India, Bangladesh, China and Indonesia, and for maize in China and the United States. Crucially, we also show that the same areas are often also at risk of high losses from pests and diseases, heat stress and to a lesser extent aridity and nutrient stress. In a solution-focussed analysis of these results, we provide a crop ideotype with tolerance of multiple stresses (including ozone) and describe how ozone effects could be included in crop breeding programmes. We also discuss altered crop management approaches that could be applied to reduce ozone impacts in the shorter term. Given the severity of ozone effects on staple food crops in areas of the world that are also challenged by other stresses, we recommend increased attention to the benefits that could be gained from addressing the ozone yield gap.

Crop quality under rising atmospheric CO2.

Crops grown under elevated CO2 (eCO2) typically exhibit enhanced yields but at the same time decreased nutritional quality. The latter effect has often been explained as a growth dilution phenomenon, but this cannot be the only process involved since crop nutrient concentrations are decreased also when production is unaffected by eCO2. We review the current knowledge on eCO2 effects on crop nutritional quality with focus on the current understanding of the possible mechanisms and processes causing these effects. Emphasis is on crop nitrogen (N) and protein concentrations but effects on other nutrients and how they compare with those on N are also covered.

Ozone pollution will compromise efforts to increase global wheat production.

Introduction of high-performing crop cultivars and crop/soil water management practices that increase the stomatal uptake of carbon dioxide and photosynthesis will be instrumental in realizing the United Nations Sustainable Development Goal (SDG) of achieving food security. To date, however, global assessments of how to increase crop yield have failed to consider the negative effects of tropospheric ozone, a gaseous pollutant that enters the leaf stomatal pores of plants along with carbon dioxide, and is increasing in concentration globally, particularly in rapidly developing countries. Earlier studies have simply estimated that the largest effects are in the areas with the highest ozone concentrations. Using a modelling method that accounts for the effects of soil moisture deficit and meteorological factors on the stomatal uptake of ozone, we show for the first time that ozone impacts on wheat yield are particularly large in humid rain-fed and irrigated areas of major wheat-producing countries (e.g. United States, France, India, China and Russia). Averaged over 2010-2012, we estimate that ozone reduces wheat yields by a mean 9.9% in the northern hemisphere and 6.2% in the southern hemisphere, corresponding to some 85 Tg (million tonnes) of lost grain. Total production losses in developing countries receiving Official Development Assistance are 50% higher than those in developed countries, potentially reducing the possibility of achieving UN SDG2. Crucially, our analysis shows that ozone could reduce the potential yield benefits of increasing irrigation usage in response to climate change because added irrigation increases the uptake and subsequent negative effects of the pollutant. We show that mitigation of air pollution in a changing climate could play a vital role in achieving the above-mentioned UN SDG, while also contributing to other SDGs related to human health and well-being, ecosystems and climate change.

A framework for assessing urban greenery's effects and valuing its ecosystem services.

Ongoing urban exploitation is increasing pressure to transform urban green spaces, while there is increasing awareness that greenery provides a range of important benefits to city residents. In efforts to help resolve associated problems we have developed a framework for integrated assessments of ecosystem service (ES) benefits and values provided by urban greenery, based on the ecosystem service cascade model. The aim is to provide a method for assessing the contribution to, and valuing, multiple ES provided by urban greenery that can be readily applied in routine planning processes. The framework is unique as it recognizes that an urban greenery comprises several components and functions that can contribute to multiple ecosystem services in one or more ways via different functional traits (e.g. foliage characteristics) for which readily measured indicators have been identified. The framework consists of five steps including compilation of an inventory of indicator; application of effectivity factors to rate indicators' effectiveness; estimation of effects; estimation of benefits for each ES; estimation of the total ES value of the ecosystem. The framework was applied to assess ecosystem services provided by trees, shrubs, herbs, birds, and bees, in green areas spanning an urban gradient in Gothenburg, Sweden. Estimates of perceived values of ecosystem services were obtained from interviews with the public and workshop activities with civil servants. The framework is systematic and transparent at all stages and appears to have potential utility in the existing spatial planning processes.

A unifying explanation for variation in ozone sensitivity among woody plants.

Tropospheric ozone is considered the most detrimental air pollutant for vegetation at the global scale, with negative consequences for both provisioning and climate regulating ecosystem services. In spite of recent developments in ozone exposure metrics, from a concentration-based to a more physiologically relevant stomatal flux-based index, large-scale ozone risk assessment is still complicated by a large and unexplained variation in ozone sensitivity among tree species. Here, we explored whether the variation in ozone sensitivity among woody species can be linked to interspecific variation in leaf morphology. We found that ozone tolerance at the leaf level was closely linked to leaf dry mass per unit leaf area (LMA) and that whole-tree biomass reductions were more strongly related to stomatal flux per unit leaf mass (r2  = 0.56) than to stomatal flux per unit leaf area (r2  = 0.42). Furthermore, the interspecific variation in slopes of ozone flux-response relationships was considerably lower when expressed on a leaf mass basis (coefficient of variation, CV = 36%) than when expressed on a leaf area basis (CV = 66%), and relationships for broadleaf and needle-leaf species converged when using the mass-based index. These results show that much of the variation in ozone sensitivity among woody plants can be explained by interspecific variation in LMA and that large-scale ozone impact assessment could be greatly improved by considering this well-known and easily measured leaf trait.

Current surface ozone concentrations significantly decrease wheat growth, yield and quality.

Tropospheric ozone is known to adversely affect crops and other vegetation. Most studies have focussed on the effects of elevated ozone levels vs. present ambient. We investigated the effect of present ambient surface ozone (O3) concentrations vs. preindustrial on a range of agronomically important response variables in field-grown wheat, using results from 33 experiments (representing 9 countries, 3 continents, 17 cultivars plus one set of 4 cultivars) having both charcoal filtered (CF) and non-filtered (NF) air treatments. Average filtration efficiency was 62%, reducing the O3 concentration from 35.6±10.6SDppb in NF to 13.7±8.8SDppb in CF. Average CF concentrations were in the range of levels believed to represent pre-industrial conditions, while NF concentrations were 7% lower than in the ambient air at plant height on the experimental sites. NF had significant (p<0.05) negative effects compared to CF on grain yield (-8.4%), grain mass (-3.7%), harvest index (-2.4%), total above-ground biomass (-5.4%), starch concentration (-3.0%), starch yield (-10.9%), and protein yield (-6.2%). No significant effect was found for grain number and protein concentration. There was a significant relationship between the effect of filtration on grain yield and the difference in O3 concentration between NF and CF treatments. The average yield loss per ppb O3 removed was 0.38% and did not systematically vary with year of experiment (ranging from 1982 to 2010) or with the average O3 level in the experiments. Although there are many differences among the field experiments included in this meta-analysis (e.g. genotype, degree of O3 pollution of the site and year, nutrient and soil condition, filtration efficiency), our study clearly shows that there is a consistent and significant effect of present ambient O3 exposure on a range of important response variables in wheat, the most strongly affected being starch yield.

The relationship between birch pollen, air pollution and weather types and their effect on antihistamine purchase in two Swedish cities.

Exposure to elevated air pollution levels can aggravate pollen allergy symptoms. The aim of this study was to investigate the relationships between airborne birch (Betula) pollen, urban air pollutants NO2, O3 and PM10 and their effects on antihistamine demand in Gothenburg and Malmö, Sweden, 2006-2012. Further, the influence of large-scale weather pattern on pollen-/pollution-related risk, using Lamb weather types (LWTs), was analysed. Daily LWTs were obtained by comparing the atmospheric pressure over a 16-point grid system over southern Sweden (scale ~3000 km). They include two non-directional types, cyclonic (C) and anticyclonic (A) and eight directional types depending on the wind direction (N, NE, E…). Birch pollen levels were exceptionally high under LWTs E and SE in both cities. Furthermore, LWTs with dry and moderately calm meteorological character (A, NE, E, SE) were associated with strongly elevated air pollution (NO2 and PM10) in Gothenburg. For most weather situations in both cities, simultaneously high birch pollen together with high air pollution had larger over-the-counter (OTC) sales of antihistamines than situations with high birch pollen alone. LWTs NE, E, SE and S had the highest OTC sales in both cities. In Gothenburg, the city with a higher load of both birch pollen and air pollution, the higher OTC sales were especially obvious and indicate an increased effect on allergic symptoms from air pollution. Furthermore, Gothenburg LWTs A, NE, E and SE were associated with high pollen and air pollution levels and thus classified as high-risk weather types. In Malmö, corresponding high-risk LWTs were NE, E, SE and S. Furthermore, occurrence of high pollen and air pollutants as well as OTC sales correlated strongly with vapour pressure deficit and temperature in Gothenburg (much less so in Malmö). This provides evidence that the combination of meteorological properties associated with LWTs can explain high levels of birch pollen and air pollution. Our study shows that LWTs represent a useful tool for integrated daily air quality forecasting/warning.

Fertilizer efficiency in wheat is reduced by ozone pollution.

Inefficient use of fertilizers by crops increases the risk of nutrient leaching from agro-ecosystems, resulting in economic loss and environmental contamination. We investigated how ground-level ozone affects the efficiency by which wheat used applied nitrogen (N) fertilizer to produce grain protein (NEP, N efficiency with respect to protein yield) and grain yield (NEY, N efficiency with respect to grain yield) across a large number of open-top chamber field experiments. Our results show significant negative ozone effects on NEP and NEY, both for a larger data set obtained from data mining (21 experiments, 70 treatments), and a subset of data for which stomatal ozone flux estimates were available (7 experiments, 22 treatments). For one experiment, we report new data on N content of different above-ground plant fractions as well as grain K and P content. Our analysis of the combined dataset demonstrates that the grain yield return for a certain investment in N fertilizer is reduced by ozone. Results from the experiment with more detailed data further show that translocation of accumulated N from straw and leaves to grains is significantly and negatively affected by ozone, and that ozone decreases fertilizer efficiency also for K and P. As a result of lower N fertilization efficiency, ozone causes a risk of increased N losses from agroecosystems, e.g. through nitrate leaching and nitrous oxide emissions, a hitherto neglected negative effect of ozone. This impact of ozone on the N cycle implies that society is facing a dilemma where it either (i) accepts increased N pollution and counteracts ozone-induced yield reductions by increasing fertilization or (ii) counteracts N pollution under elevated ozone by reducing fertilization, accepting further yield loss adding to the direct effect of ozone on yield.

Epidemiological analysis of ozone and nitrogen impacts on vegetation - Critical evaluation and recommendations.

For human health studies, epidemiology has been established as important tool to examine factors that affect the frequency and distribution of disease, injury, and other health-related events in a defined population, serving the purpose of establishing prevention and control programs. On the other hand, gradient studies have a long tradition in the research of air pollution effects on plants. While there is no principal difference between gradient and epidemiological studies, the former address more one-dimensional transects while the latter focus more on populations and include more experience in making quantitative predictions, in dealing with confounding factors and in taking into account the complex interplay of different factors acting at different levels. Epidemiological analyses may disentangle and quantify the contributions of different predictor variables to an overall effect, e.g. plant growth, and may generate hypotheses deserving further study in experiments. Therefore, their use in ecosystem research is encouraged. This article provides a number of recommendations on: (1) spatial and temporal aspects in preparing predictor maps of nitrogen deposition, ozone exposure and meteorological covariates; (2) extent of a dataset required for an analysis; (3) choice of the appropriate regression model and conditions to be satisfied by the data; (4) selection of the relevant explanatory variables; (5) treatment of interactions and confounding factors; and (6) assessment of model validity.

Influence of urban vegetation on air pollution and noise exposure - A case study in Gothenburg, Sweden.

Air pollution levels (NO2, PAHs, O3) were investigated, before (BLE) and after (ALE) leaf emergence, in the urban landscape of Gothenburg, Sweden. The aims were to study the 1) spatial and temporal variation in pollution levels between urban green areas, 2) effect of urban vegetation on air pollution levels at the same distance from a major emission source (traffic route), 3) improvement of urban air quality in urban parks compared to adjacent sites near traffic, 4) correlation between air pollution and noise in a park. O3 varied little over the urban landscape. NO2 varied strongly and was higher in situations strongly influenced by traffic. Four PAH variables were included: total PAH, total particle-bound PAH, the quantitatively important gaseous phenanthrene and the highly toxic particle-bound benzo(a)pyrene. The variation of PAHs was similar to NO2, but for certain PAHs the difference between highly and less polluted sites was larger than for NO2. At a vegetated site, NO2 and particulate PAH levels were lower than at a non-vegetated site at a certain distance from a busy traffic route. This effect was significantly larger ALE compared to BLE for NO2, indicating green leaf area to be highly significant factor for air quality improvement. For particulate PAHs, the effect was similar BLE and ALE, indicating that tree bark and branches also could be an important factor in reducing air pollution. Parks represented considerably cleaner local environments (park effect), which is likely to be a consequence of both a dilution (distance effect) and deposition. Noise and air pollution (NO2 and PAH) levels were strongly correlated. Comparison of noise levels BLE and ALE also showed that the presence of leaves significantly reduced noise levels. Our results are evidence that urban green spaces are beneficial for urban environmental quality, which is important to consider in urban planning.

Past, present and future concentrations of ground-level ozone and potential impacts on ecosystems and human health in northern Europe.

This review summarizes new information on the current status of ground-level ozone in Europe north of the Alps. There has been a re-distribution in the hourly ozone concentrations in northern Europe during 1990-2015. The highest concentrations during summer daytime hours have decreased while the summer night-time and winter day- and night-time concentrations have increased. The yearly maximum 8-h mean concentrations ([O3]8h,max), a metric used to assess ozone impacts on human health, have decreased significantly during 1990-2015 at four out of eight studied sites in Fennoscandia and northern UK. Also the annual number of days when the yearly [O3]8h,max exceeded the EU Environmental Quality Standard (EQS) target value of 60ppb has decreased. In contrast, the number of days per year when the yearly [O3]8h,max exceeded 35ppb has increased significantly at two sites, while it decreased at one far northern site. [O3]8h,max is predicted not to exceed 60ppb in northern UK and Fennoscandia after 2020. However, the WHO EQS target value of 50ppb will still be exceeded. The AOT40 May-July and AOT40 April-September metrics, used for the protection of vegetation, have decreased significantly at three and four sites, respectively. The EQS for the protection of forests, AOT40 April-September 5000ppbh, is projected to no longer be exceeded for most of northern Europe sometime before the time period 2040-2059. However, if the EQS is based on Phytotoxic Ozone Dose (POD), POD1, it may still be exceeded by 2050. The increasing trend for low and medium range ozone concentrations in combination with a decrease in high concentrations indicate that a new control strategy, with a larger geographical scale than Europe and including methane, is needed for ozone abatement in northern Europe.

Differential effects of ozone on photosynthesis of winter wheat among cultivars depend on antioxidative enzymes rather than stomatal conductance.

Five modern cultivars of winter wheat (Triticum aestivum L.): Yangmai16 (Y16), Yangmai 15 (Y15), Yangfumai 2 (Y2), Yannong 19 (Y19) and Jiaxing 002 (J2) were investigated to determine the impacts of elevated ozone concentration (E-O3) on photosynthesis-related parameters and the antioxidant system under fully open-air field conditions in China. The plants were exposed to E-O3 at 1.5 times the ambient ozone concentration (A-O3) from the initiation of tillering to final harvest. Pigments, gas exchange rates, chlorophyll a fluorescence, antioxidants contents, antioxidative enzyme activity and lipid oxidation were measured in three replicated plots throughout flag leaf development. Results showed that significant O3 effects on most variables were only found during the mid-grain filling stage. Across five cultivars, E-O3 significantly accelerated leaf senescence, as indicated by increased lipid oxidation as well as faster declines in pigment amounts and photosynthetic rates. The lower photosynthetic rates were mainly due to non-stomatal factors, e.g. lower maximum carboxylation capacity and electron transport rates. There were strong interactions between O3 and cultivar in photosynthetic pigments, light-saturated photosynthesis rate and chlorophyll a fluorescence with O3-sensitive (Y19, Y2 and Y15) and O3-tolerant (J2, Y16) cultivars being clearly differentiated in their responses to E-O3. E-O3 significantly influenced the antioxidative enzymes but not antioxidant contents. Significant interactions between O3 and cultivar were found in antioxidative enzymes, such as SOD and CAT, but not in stomatal conductance (gs). Therefore, it can be concluded that antioxidative enzymes rather than gs or antioxidants are responsible for the differential responses to E-O3 among cultivars. These findings provide important information for the development of accurate modeling O3 effects on crops, especially with respect to the developmental stage when O3 damage to photosynthesis becomes manifest.