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Background The paper presents an overview of air quality in the 27 member countries of the European Union (EU) and the United Kingdom (previous EU-28), from 2000 to 2017. We reviewed the progress made towards meeting the air quality standards established by the EU Ambient Air Quality Directives (European Council Directive 2008/50/EC) and the World Health Organization (WHO) Air Quality Guidelines by estimating the trends (Mann-Kendal test) in national emissions of main air pollutants, urban population exposure to air pollution, and in mortality related to exposure to ambient fine particles (PM2.5) and tropospheric ozone (O3). Results Despite significant reductions of emissions (e.g., sulfur oxides: ~ 80%, nitrogen oxides: ~ 46%, non-methane volatile organic compounds: ~ 44%, particulate matters with a diameter lower than 2.5 µm and 10 µm: ~ 30%), the EU-28 urban population was exposed to PM2.5 and O3 levels widely exceeding the WHO limit values for the protection of human health. Between 2000 and 2017, the annual PM2.5-related number of deaths decreased (- 4.85 per 106 inhabitants) in line with a reduction of PM2.5 levels observed at urban air quality monitoring stations. The rising O3 levels became a major public health issue in the EU-28 cities where the annual O3-related number of premature deaths increased (+ 0.55 deaths per 106 inhabitants). Conclusions To achieve the objectives of the Ambient Air Quality Directives and mitigate air pollution impacts, actions need to be urgently taken at all governance levels. In this context, greening and re‐naturing cities and the implementation of fresh air corridors can help meet air quality standards, but also answer to social needs, as recently highlighted by the COVID-19 lockdowns.

Ground-level ozone (O3) is the main phytotoxic air pollutant causing crop yield reduction in China. As the main grain producing area in China, the Yangtze River Delta (YRD) is facing serious O3 pollution. This study analyzed the hourly ground-level O3 observation data of 158 stations from 2014 to 2019 in YRD, and grain production data of 193 districts and counties. The exposure-response relationships based on AOT40 (accumulated hourly O3 concentration above 40 ppb) was used to estimate the yield loss and economic loss of two food crops (winter wheat and rice). This study used spatial interpolation and calculated the specific data values of each district and county in order to improve the assessment reliability. For years 2014–2019, averaged O3 concentration during the 75 days growing period of rice and wheat were 33.1–50.6 ppb and 32.2–48.0 ppb, AOT40 value were 5.2–12.0 ppm h and 4.6–9.4 ppm h, and the averaged relative yield losses were 4.9%–11.4% and 9.4%–19.3%, respectively. The trend of O3 in the YRD in a six-year period peaked in 2016 and 2017 for rice and winter wheat, respectively. During 2014–2017, the average estimated yield loss of rice was 2445 Mt. accounting for about 9.1% of the actual production, and the average estimated economic loss was about 1037 million USD; for winter wheat, it was 2025 Mt, 20.4% and 736 million USD, respectively. These results urge governments to provide effective policies and measures to control O3 pollution.

Maize is the second most important crop per harvested area in the world. The North China Plain (NCP) is a highly populated and relevant agricultural region in China, experiencing some of the highest ozone (O3) concentrations worldwide. It produces ~24% of the total maize production of China in years 2014–2017. For these years, we used observational O3 data in combination with geostatistic methods to estimate county-level production and economic losses due to O3 in the NCP. AOT40 (accumulated ozone exposure over an hourly threshold of 40 ppb) values during the maize growing season (90 days before maturity) progressively increased in the four consecutive years: 13.7 ppm h, 15.4 ppm h, 16.9 ppm h and 22.7 ppm h. Mean relative yield losses were 8.2% in 2014, 9.2% in 2015, 10.4% in 2016 and 13.4% in 2017. These yield losses, derived from exposure-response functions, resulted in crop production losses of 530.3 × 104 t, 617.8 × 104 t, 713.8 × 104 t, and 953.4 × 104 t, as well as economic losses of 2343 million USD, 2672 million USD, 1887 million USD, and 2404 million USD from 2014 to 2017. The NCP is a key area in China for monitoring the effectiveness of the clean air action policies aiming at reducing emissions of air pollutants. Despite these measures, O3 concentrations have increased in NCP, and reduction of this pollutant are challenging. We suggest an increase in the number of rural air quality stations for better characterizing O3 trends in cropland areas, as well as the application of different mitigation measures. They may involve more stringent air quality regulations and changes in crops, breeding tolerant cultivars and a crop management taking into account O3 pollution.

Ozone (O3) pollution can induce changes in plant growth and metabolism, and in turn, affects isoprene emission (ISO), but the extent of these effects may be modified by co-occurring soil water and nitrogen (N) availability. To date, however, much less is known about the combined effects of two of these factors on isoprene emission from plants. We investigated for the first time the combined effects of O3 exposure (CF, charcoal-filtered air; EO3, non-filtered air plus 40 ppb of O3), N addition (N0, no additional N; N50, 50 kg ha−1 year−1 of N) and moderate drought (WW, well-watered; WR, 40% of WW irrigation) on photosynthetic carbon assimilation and ISO emission in hybrid poplar at both leaf- and plant-level over time. Consistent with leaf-level photosynthesis (Pnleaf) and ISO (ISOleaf) responses, plant-level ISO (ISOplant) responses to O3, N addition and moderate drought were more marked after long exposure (September) than short exposure duration (July). EO3 significantly decreased ISOleaf and Pnleaf, while WR and N50 significantly increased them. Although O3 and water interacted significantly to affect Pnleaf over the exposure duration, neither N50 nor WR mitigated the negative effects of EO3 on ISOleaf. When ISO was scaled up to the plant level, the WR-induced increase in ISOleaf under EO3 was offset by a reduction in total leaf area. By contrast, effects of EO3 on ISOplant were not changed by N addition. Our results highlight that the dynamic effects on ISO emission change over the exposure duration depending on involved co-occurring factors and evaluation scales.

Wheat is a major staple food and its sensitivity to the gas pollutant ozone (O3) depends on the cultivar. However, few chamber-less studies assessed current ambient O3 effects on a large number of wheat cultivars. In this study, we used ethylenediurea (EDU), an O3 protectant whose protection mechanisms are still unclear, to test photosynthetic pigments, gas exchange, antioxidants, and yield of 15 cultivars exposed to 17.4 ppm h AOT40 (accumulated O3 over an hourly concentration threshold of 40 ppb) over the growing season at Beijing suburb, China. EDU significantly increased light-saturated photosynthesis rate (Asat), photosynthetic pigments (i.e., chlorophyll and carotenoid), and total antioxidant capacity, while reduced malondialdehyde and reduced ascorbate contents. In comparison with EDU-treated plants (control), plants treated with water (no protection from ambient O3) significantly decreased yield, weight of 1000 grains, and harvest index by 20.3%, 15.1%, and 14.2%, respectively, across all cultivars. There was a significant interaction between EDU and cultivars in all tested variables with exception of Asat, chlorophyll, and carotenoid. The cultivar-specific sensitivity to O3 was ranked from highly sensitive (> 25% change) to less sensitive (

Five winter wheat cultivars were exposed to ambient (A-O3) and elevated (E-O3, 1.5 ambient) O3 in a fully open-air fumigation system in China. Ozone exposure- and flux based response relationships were established for seven physiological variables related to photosynthesis. The performance of the fitting of the regressions in terms of R2 increased when second order regressions instead of first order ones were used, suggesting that effects of O3 were more pronounced towards the last developmental stages of the wheat. The more robust indicators were those related with CO2 assimilation, Rubisco activity and RuBP regeneration capacity (Asat, Jmax and Vcmax), and chlorophyll content (Chl). Flux-based metrics (PODy, Phytotoxic O3 Dose over a threshold y nmol O3 m− 2 s− 1) predicted slightly better the responses to O3 than exposure metrics (AOTX, Accumulated O3 exposure over an hourly Threshold of X ppb) for most of the variables. The best performance was observed for metrics POD1 (Asat, Jmax and Vcmax) and POD3 (Chl). For this crop, the proposed response functions could be used for O3 risk assessment based on physiological effects and also to include the influence of O3 on yield or other variables in models with a photosynthetic component.

The effects of elevated ozone on C (carbon), N (nitrogen) and P (phosphorus) ecological stoichiometry and nutrient resorption in different organs including leaves, stems and roots were investigated in poplar clones 546 (P. deltoides cv. '55/56' × P. deltoides cv. 'Imperial') and 107 (P. euramericana cv. '74/76') with a different sensitivity to ozone. Plants were exposed to two ozone treatments, NF (non-filtered ambient air) and NF60 (NF with targeted ozone addition of 60 ppb), for 96 days in open top chambers (OTCs). Significant ozone effects on most variables of C, N and P ecological stoichiometry were found except for the C concentration and the N/P in different organs. Elevated ozone increased both N and P concentrations of individual organs while for C/N and C/P ratios a reduction was observed. On these variables, ozone had a greater effect for clone 546 than for clone 107. N concentrations of different leaf positions ranked in the order upper > middle > lower, showing that N was transferred from the lower senescent leaves to the upper ones. This was also indicative of N resorption processes, which increased under elevated ozone. N resorption of clone 546 was 4 times larger than that of clone 107 under ambient air (NF). However, elevated ozone (NF60) had no significant effect on P resorption for both poplar clones, suggesting that their growth was only limited by N, while available P in the soil was enough to sustain growth. Understanding ecological stoichiometric responses under ozone stress is crucial to predict future effects on ecological processes and biogeochemical cycles.

The relationships of CO2 assimilation under saturated-light conditions (Asat) with exposure- (AOTX, Accumulated Ozone exposure over a hourly Threshold of X ppb) and flux-based (PODY, Phytotoxic Ozone Dose over a hourly threshold Y nmol·m-2·s-1) O3 metrics was studied on three common urban trees, Fraxinus chinensis (FC), Platanus orientalis (PO) and Robinia pseudoacacia (RP). Parameterizations for a stomatal multiplicative model were proposed for the three species. RP was the species showing lower species-specific maximum stomatal conductance (gmax) and experiencing lower cumulative O3 uptake along the experiment, but in contrast it was the most sensitive to O3. PODY was slightly better than AOTX metric at estimating relative Asat (R-Asat)for PO and RB but not for FC. The best fittings obtained for the regressions between R-Asat and AOTX for FC, PO and RP were 0.904, 0.868, and 0.876, when the thresholds of X were 60ppb, 55ppb and 30ppb, respectively. However, AOT40 performed also well for all of them, with R2 always >0.83. For PODY, the highest R2 values for FC, PO and RB were 0.863, 0.897 and 0.911 at thresholds Y=7, 5 and 1nmolO3m-2s-1, respectively. Given the potentially higher O3 removal capacity of FC and PO by stomatal uptake and their lower sensitivity to this pollutant than RP, the former two species would be appropriate for urban gardens and areas where O3 levels are high. Parameterization and modeling of stomatal conductance for the main urban tree species may provide reliable estimations of the stomatal uptake of O3 and other gaseous pollutants by vegetation, which may support decision making on the most suitable species for green urban planning in polluted areas.

Ground-level ozone (O3) is an important phytotoxic air pollutant in China. In order to compare the sensitivity of common poplar clones to O3 in China and explore the possible mechanism, five poplar clones, clone DQ (Populus cathayana), clone 84 K (P. alba × P. glandulosa), clone WQ156 (P. deltoids × P. cathayana), clone 546 (P. deltoides cv. ‘55/56’ × P. deltoides cv. ‘Imperial’) and clone 107 (P. euramericana cv. ‘74/76’) were exposed to four O3 treatments. According to the date of the initial visible O3 symptom and the slopes of O3 exposure-response relationships with the relative light-saturated rate of CO2 assimilation, we found that clone DQ and clone 546 were the most sensitive to O3, clone 84 K and clone WQ156 were the less sensitive, and clone 107 was the most tolerant, which could provide a basis to select O3 tolerant clones for poplar planting at areas with serious O3 pollution. Elevated O3 significantly reduced photosynthetic parameters, total phenols content, potential antioxidant capacity, leaf mass per area and biomass of five poplar clones, and there were significant interactions between O3 and clones for most photosynthetic parameters. Elevated O3 also significantly increased malondialdehyde content and total ascorbate content. The responses of total antioxidant capacity for poplar clones to elevated O3 were different, as indicated by the increase for clone 107 and reduction for other clones under elevated O3 treatment. Our results on the sensitivity of different poplar clones to O3 are not related to leaf stomatal conductance, leaf constitutive antioxidant levels or leaf morphology of plant grown in clean air. The possible reason is little difference in leaf traits among clones within close species, suggesting that more properties of plants should be considered for exploring the sensitivity mechanism of close species, such as mesophyll conductance, antioxidant enzyme activity and apoplastic antioxidants.

Since the Industrial Revolution, the global ambient O3 concentration has more than doubled. Negative impact of O3 on some common crops such as wheat and soybeans has been widely recognized, but there is relatively little information about maize, the typical C4 plant and third most important crop worldwide. To partly compensate this knowledge gap, the maize cultivar (Zhengdan 958, ZD958) with maximum planting area in China was exposed to a range of chronic ozone (O3) exposures in open top chambers (OTCs). The O3 effects on this highly important crop were estimated in relation to two O3 metrics, AOT40 (accumulated hourly O3 concentration over a threshold of 40 ppb during daylight hours) and POD6 (Phytotoxic O3 Dose above a threshold flux of 6 nmol O3 m−2 s−1 during a specified period). We found that (1) the reduced light-saturated net photosynthetic rate (Asat) mainly caused by non-stomatal limitations across heading and grain filling stages, but the stomatal limitations at the former stage were stronger than those at the latter stage; (2) impact of O3 on water use efficiency (WUE) of maize was significantly dependent on developmental stage; (3) yield loss induced by O3 was mainly due to a reduction in kernels weight rather than in the number of kernels; (4) the performance of AOT40 and POD6 was similar, according to their determination coefficients (R2); (5) the order of O3 sensitivity among different parameters was photosynthetic parameters > biomass parameters > yield-related parameters; (6) Responses of Asat to O3 between heading and gran filling stages were significantly different based on AOT40 metric, but not POD6. The proposed O3 metrics-response relationships will be valuable for O3 risk assessment in Asia and also for crop productivity models including the influence of O3.

The effect of lockdown due to coronavirus disease (COVID-19) pandemic on air pollution in four Southern European cities (Nice, Rome, Valencia and Turin) and Wuhan (China) was quantified, with a focus on ozone (O3). Compared to the same period in 2017–2019, the daily O3 mean concentrations increased at urban stations by 24% in Nice, 14% in Rome, 27% in Turin, 2.4% in Valencia and 36% in Wuhan during the lockdown in 2020. This increase in O3 concentrations is mainly explained by an unprecedented reduction in NOx emissions leading to a lower O3 titration by NO. Strong reductions in NO2 mean concentrations were observed in all European cities, ~53% at urban stations, comparable to Wuhan (57%), and ~65% at traffic stations. NO declined even further, ~63% at urban stations and ~78% at traffic stations in Europe. Reductions in PM2.5 and PM10 at urban stations were overall much smaller both in magnitude and relative change in Europe (~8%) than in Wuhan (~42%). The PM reductions due to limiting transportation and fuel combustion in institutional and commercial buildings were partly offset by increases of PM emissions from the activities at home in some of the cities. The NOx concentrations during the lockdown were on average 49% lower than those at weekends of the previous years in all cities. The lockdown effect on O3 production was ~10% higher than the weekend effect in Southern Europe and 38% higher in Wuhan, while for PM the lockdown had the same effect as weekends in Southern Europe (~6% of difference). This study highlights the challenge of reducing the formation of secondary pollutants such as O3 even with strict measures to control primary pollutant emissions. These results are relevant for designing abatement policies of urban pollution., Graphical abstract Unlabelled Image, Highlights • Air quality during the COVID-19 lockdown in 4 European and 1 Chinese cities • The lockdown caused a substantial reduction in NOx in all cities (~ 56%) • Reductions in PM were much higher in Wuhan (~ 42%) than in Europe (~ 8%) • The lockdown caused an ozone increase in all cities (17% in Europe, 36% in Wuhan) • The lockdown effect on O3 production was higher than the weekend effect

Tropospheric ozone (O3) is a pollutant of widespread concern in the world and especially in China for its negative effects on agricultural crops. For the first time, yield and economic losses of wheat between 2014 and 2017 were estimated for the North China Plain (NCP) using observational hourly O3 data from 312 monitoring stations and exposure-response functions based on AOT40 index (accumulated hourly O3 concentration above 40 ppb) from a Chinese study. AOT40 values from 2014 to 2017 during the wheat growing seasons (75-days, 44 before and 30 after mid-anthesis) ranged from 3.1 to 14.9 ppm h, 4.9–17.5 ppm h, 7.3–17.6 ppm h, and 0.5–18.6 ppm h, respectively. The highest AOT40 values were observed in the Beijing-Tianjin-Hebei region. The values of relative yield losses from 2014 to 2017 were in the ranges of 6.4–30.5%, 10.0–35.8%, 14.9–34.1%, and 21.6–38.2%, respectively. The total wheat production losses in NCP for 2014–2017 accounted for 18.5%, 22.7%, 26.2% and 30.8% in the whole production, while the economic losses amounted to 6,292 million USD, 8,524 million USD, 10,068 million USD, and 12,404 million USD, respectively. The important impact of O3 in this area, which is of global importance, should be considered when assessing wheat yield production. Our results also show an increasing trend in AOT40, relative yield loss, total crop production loss and economic loss in the four consecutive years.

The effects of current and future elevated O3 concentrations (e[O3]) were investigated by a meta-analysis for poplar, a widely distributed genus in the Northern Hemisphere with global economic importance. Current [O3] has significantly reduced CO2 assimilation rate (Pn) by 33% and total biomass by 4% in comparison with low O3 level (charcoal-filtered air, CF). Relative to CF, an increase in future [O3] would further enhance the reduction in total biomass by 24%, plant height by 17% and plant leaf area by 19%. Isoprene emissions could decline by 34% under e[O3], with feedback implications in reducing the formation of secondary air pollutants including O3. Reduced stomatal conductance and lower foliar area might increase runoff and freshwater availability in O3 polluted areas. Higher cumulated O3 exposure over a threshold of 40 ppb (AOT40) induced larger reductions in Pn, total biomass and isoprene emission. Relationships of light-saturated photosynthesis rates (Asat), total biomass and chlorophyll content with AOT40 using a global dataset are provided. These relationships are expected to improve O3 risk assessment and also to support the inclusion of the effect of O3 in models addressing plantation productivity and carbon sink capacity.

Skin affections after sulfur mustard (SM) exposure include erythema, blister formation and severe inflammation. An antidote or specific therapy does not exist. Anti-inflammatory compounds as well as substances counteracting SM-induced cell death are under investigation. In this study, we investigated the benzylisoquinoline alkaloide berberine (BER), a metabolite in plants like berberis vulgaris, which is used as herbal pharmaceutical in Asian countries, against SM toxicity using a well-established in vitro approach. Keratinocyte (HaCaT) mono-cultures (MoC) or HaCaT/THP-1 co-cultures (CoC) were challenged with 100, 200 or 300 mM SM for 1 h. Post-exposure, both MoC and CoC were treated with 10, 30 or 50 mu M BER for 24 h. At that time, supernatants were collected and analyzed both for interleukine (IL) 6 and 8 levels and for content of adenylate-kinase (AK) as surrogate marker for cell necrosis. Cells were lysed and nucleosome formation as marker for late apoptosis was assessed. In parallel, AK in cells was determined for normalization purposes. BER treatment did not influence necrosis, but significantly decreased apoptosis. Anti-inflammatory effects were moderate, but also significant, primarily in CoC. Overall, BER has protective effects against SM toxicity in vitro. Whether this holds true should be evaluated in future in vivo studies.

Four genotypes of snap bean (Phaseolus vulgaris L.) were selected to study the effects of ambient ozone concentration at a cropland area around Beijing by using 450 ppm of ethylenediurea (EDU) as a chemical protectant. During the growing season, the 8 h (9:00–17:00) average ozone concentration was very high, approximately 71.3 ppb, and AOT40 was 29.0 ppm.h. All genotypes showed foliar injury, but ozone-sensitive genotypes exhibited much more injury than ozone-tolerant ones. Compared with control, EDU significantly alleviated foliar injury, increased photosynthesis rate and chlorophyll a fluorescence, Vcmax and Jmax, and seed and pod weights in ozone-sensitive genotypes but not in ozone-tolerant genotypes. EDU did not significantly affect antioxidant contents in any of the genotypes. Therefore, EDU effectively protected sensitive genotypes from ambient ozone damage, while protection on ozone-tolerant genotypes was limited. EDU can be regarded as a useful tool in risk assessment of ambient ozone on food security.

Poplar clones 546 (P. deltoides cv. '55/56'×P. deltoides cv. 'Imperial') and 107 (P. euramericana cv. '74/76') were exposed to five ozone concentrations in 15 open-top chambers (OTCs). Both ozone exposure (AOT40, Accumulation Over a Threshold hourly ozone concentration of 40ppb) and flux-based (POD7, Phytotoxic Ozone Dose above an hourly flux threshold of 7nmol O3 m-2 PLA (projected leaf area) s-1) response relationships were established with photosynthesis, leaf morphology and biomass variables. Increases in both metrics showed significant negative relationships with light-saturated photosynthesis rate, chlorophyll content, leaf mass per area, actual photochemical efficiency of PSII in the light and root biomass but not with stomatal conductance (gs), leaf and stem biomass. Ozone had a greater impact on belowground than on aboveground biomass. The ranking of these indicators from higher to lower sensitivity to ozone was: photosynthetic parameters, morphological index, and biomass. Clone 546 had a higher sensitivity to ozone than clone 107. The coefficients of determination (R2) were similar between exposure- and flux-based dose-response relationships for each variable. The critical levels (CLs) for a 5% reduction in total biomass for the two poplar clones were 14.8ppmh for AOT40 and 9.8mmol O3 m-2 PLA for POD7. In comparison, equivalent reduction occurred at much lower values in photosynthetic parameters (4ppmh for AOT40 and 3mmol O3 m-2 PLA for POD7) and LMA (5.8ppmh for AOT40 and 4mmol O3 m-2 PLA for POD7). While in recent decades different CLs have been proposed for several plant receptors especially in Europe, studies focusing on both flux-based dose-response relationships and CLs are still scarce in Asia. This study is therefore valuable for regional O3 risk assessment in Asia.

Despite the high ozone levels measured in China, and in Beijing in particular, reports of ozone-induced visible injury in vegetation are very scarce. Visible injury was investigated on July and August 2013 in the main parks, forest and agricultural areas of Beijing. Ozone injury was widespread in the area, being observed in 28 different species. Symptoms were more frequent in rural areas and mountains from northern Beijing, downwind from the city, and less frequent in city gardens. Among crops, injury to different types of beans (genera Phaseolus, Canavalia and Vigna) was common, and it was also observed in watermelon, grape vine, and in gourds. Native species such as ailanthus, several pines and ash species were also symptomatic. The black locust, the rose of Sharon and the Japanese morning glory were among the injured ornamental plants. Target species for broader bio-monitoring surveys in temperate China have been identified.

Seven experiments carried out in Italy and Spain have been used to parameterising a stomatal conductance model and establishing exposure- and dose-response relationships for yield and quality of tomato with the main goal of setting O3 critical levels (CLe). CLe with confidence intervals, between brackets, were set at an accumulated hourly O3 exposure over 40 nl l(-1), AOT40 = 8.4 (1.2, 15.6) ppm h and a phytotoxic ozone dose above a threshold of 6 nmol m(-2) s(-1), POD6 = 2.7 (0.8, 4.6) mmol m(-2) for yield and AOT40 = 18.7 (8.5, 28.8) ppm h and POD6 = 4.1 (2.0, 6.2) mmol m(-2) for quality, both indices performing equally well. CLe confidence intervals provide information on the quality of the dataset and should be included in future calculations of O3 CLe for improving current methodologies. These CLe, derived for sensitive tomato cultivars, should not be applied for quantifying O3-induced losses at the risk of making important overestimations of the economical losses associated with O3 pollution.

Current ozone (O3) levels are high enough to negatively affect vegetation and may become worse in the future. Ozone risk assessments have recently shifted from exposure-based to flux-based metrics. Modeling stomatal O3 fluxes requires hourly O3 and meteorological data, which are not always available. Large datasets of O3 concentrations measured with passive samplers exist worldwide, and usually provide weekly to monthly means. We tested the suitability of using aggregated data instead of hourly data for O3 flux calculations with 3-year time series of O3 data from 24 Spanish air quality stations. Five different approaches and three different parameterizations were tested. Ozone-averaged values in combination with hourly meteorological data provided the most robust estimates of accumulated O3 fluxes (Phytotoxic Ozone Dose with no threshold, POD0), and the median of the absolute percent error (MAPE) due to aggregation came close to 5%. Aggregations from 1 week to 1 month yielded similar errors, which is important in the cost-efficiency terms of the chosen passive sampler exposure periodicity. One major limitation of these approaches is that they are not suitable for high POD thresholds, and that accuracy of the measurements with passive samplers has to be strictly assured in order to finally obtain acceptable errors. A combination of meteorological data and O3 passive sampler measurements may be used to estimate O3 fluxes at remote forest sites as a valuable risk assessment tool. © 2016

Plant responses to enhanced ozone levels have been studied in two pairs of evergreen-deciduous species (Pistacia terebinthus vs. P. lentiscus; Viburnum lantana vs. V. tinus) in Open Top Chambers. Ozone induced widespread visible injury, significantly reduced CO(2) assimilation and stomatal conductance (g(s)), impaired Rubisco efficiency and regeneration capacity (V(c,max,)J(max)) and altered fluorescence parameters only in the deciduous species. Differences in stomatal conductance could not explain the observed differences in sensitivity. In control plants, deciduous species showed higher superoxide dismutase (SOD) activity than their evergreen counterparts, suggesting metabolic differences that could make them more prone to redox imbalances. Ozone induced increases in SOD and/or peroxidase activities in all the species, but only evergreens were able to cope with the oxidative stress. The relevancy of these results for the effective ozone flux approach and for the current ozone Critical Levels is also discussed.

The effects of ozone in four maple species, Acer campestre, A. opalus subsp. granatense, A. monspessulanum and A. pseudoplatanus were studied in OTC under two different experimental conditions: in charcoal filtered air (CF), and in non filtered air plus 30 ppb ozone (NF+30). The four species of maple showed contrasting sensitivity to ozone as demonstrated by visible injury development, gas exchange and chlorophyll a fluorescence, and growth measurements. Plant injury index (i.e. a combination of percentage of injured leaves and leaf surface affected) was more consistently related with physiological measurements than the onset of first symptom of visible injury. Differences in ozone sensitivity among species may be partly related to higher stomatal conductances in A. opalus and A. pseudoplatanus. In these two species, ozone produced significant reductions in CO2 assimilation under saturating light conditions (Asat), stomatal conductance (gs), transpiration rate (Tr) and Water Use Efficiency (WUE) (the latter also significantly declined in A. campestre) towards the end of summer, while intercellular CO2 concentrations (Ci) increased significantly. In asymptomatic leaves of A. opalus, neither stomatal limitation nor photoinhibitory damage (Fv/Fm) could explain the observed decline of Asat, and photosynthesis was down regulated by reducing the proportion of absorbed energy used in photochemistry (ΦPSII) at expenses of the energy dispersed non-photochemically (NPQ). Leaf N content also declined significantly in A. pseudoplatanus. Plants exposed to ozone showed a tendency to decrease growth, but it was not significant within the exposure period for any of the four species. The most sensitive species were A. opalus and A. pseudoplatanus, while the species with the smallest and more coriaceous leaves, A. monspessulanum, was the most resistant.

A biomonitoring study using the ozone-sensitive bioindicator plant Nicotiana tabacum cv. Bel-W3 was conducted in the city of Valencia (eastern Spain) and surrounding areas in 2002. Plants were exposed to ambient air at seven sites, including four traffic-exposed urban sites, a large urban garden and a suburban and a rural station, for six consecutive 2-week periods using highly standardised methods. Foliar injury was registered at all stations in at least one of the exposure periods. The urban stations submitted to intense traffic showed lower ozone injury than the less traffic-exposed stations. Strong changes in the intensity of ozone injury were observed for the different exposure periods. Leaf injury was significantly related to both mean ozone values (24 and 12 h means) and cumulative exposure indices (AOT20, AOT40). However, correlation strength was moderate (r s = 0.39 to 0.58), suggesting that the plant response to ozone was modified by environmental factors. The use of sensitive bioindicators like tobacco Bel-W3 in cities provides complementary information to that of continuously operating air quality monitors, as the impact of ambient ozone levels is directly measured.

[EN] An open-top chamber study was conducted to investigate the tissue and cellular-level foliar effects of ozone (O-3) on a Mediterranean evergreen species, the mastic plant (Pistacia lentiscus L.). Plants were exposed at three different O-3 levels, and leaf samples were collected periodically from the beginning of the exposure. Although no visible foliar injury was evident, alterations of the plastids and vacuoles in the mesophyll were observed. Senescence processes were accelerated with an anomalous stacking of tannin vacuoles, and a reduction in the size and number of the chloroplasts. Overall, most of the modifications induced by O-3 were consistent with previously reported observations on deciduous broadleaf species, with the exception of alterations in the cells covering the secretory channels, reported here as a new finding. Comments on the feasibility of using microscopy to validate O-3 related field observations and subtle foliar injury are also given. (C) 2004 Elsevier Ltd. All rights reserved.

In eastern Spain, studies combining the tracking and meso-scale circulations of air pollutants with the evaluation of their effects on plants have been undertaken since 1994. Meso-scale processes are very important from the point of view of how and where forest ecosystems are affected by point sources and regional air pollution in the Mediterranean area. The first results of these field surveys show that in 1994, 1995 and 1996, the distribution pattern of ozone visual injury (chlorotic mottle) in Pinus halepensis correlated with the penetration of pollutants transported by the sea-breeze into coastal valleys of Castellón (eastern Spain). In this tree species, longer needles are associated with higher chlorotic mottle, and ozone injury seems to be among the factors affecting needle retention and crown transparency.

The paper provides basic information about background, objectives and structure of O 3 SWE (Ozone at the permanent monitoring plots in South-Western Europe), an international co-operative project aimed at evaluating O 3 concentrations, cumulative exposure, uptake and effects on forest vegetation in four countries of South-Western Europe (France, Italy, Luxenbourg, Spain and Switzerland). The project covers a total of 83 permanent plots of the EU and UN/ECE intensive forest monitoring programme and span over three years of investigation (2000–2002). The O 3 SWE project aims to demonstrate how, using data collected routinely in an intensive forest monitoring network, O 3 exposure, flux and effects can be assessed and exceedances critically evaluated.

Plants of one evergreen oak (Quercus ilex) and three deciduous oaks (Q. faginea, with small leaves; Q. pyrenaica and Q. robur, with large leaves) were exposed both to filtered air and to enhanced ozone levels in Open-Top Chambers. Q. faginea and Q. pyrenaica were studied for the first time. Based on visible injury, gas exchange, chlorophyll content and biomass responses, Q. pyrenaica was the most sensitive species, and Q. ilex was the most tolerant, followed by Q. faginea. Functional leaf traits of the species were related to differences in sensitivity, while accumulated ozone flux via stomata (POD1.6) partly contributed to the observed differences. For risk assessment of Mediterranean vegetation, the diversity of responses detected in this study should be taken into account, applying appropriate critical levels. © 2010 Elsevier Ltd. All rights reserved., We thank both the Ministerio de Medio Ambiente y Medio Rural y Maritimo (in collaboration with ICP-Forests), and the Conselleria de Medi Ambient, Aigua i Habitatge and Interreg III (ForMedOzone and VegetPollOzone projects) for supporting the OTC activity. Institut Universitario CEAM-UMH is also supported by Generalitat Valenciana and Fundacion Bancaja, benefiting from CONSOLIDER-INGENIO 2010 (GRACCIE) and Prometeo (Generalitat Valenciana) Programs. Filippo Bussotti and two anonymous referees are thanked for their useful comments. Carmen Martin is also thanked for taking care of the plants.

Ozone effects on the perennial forb Lamottea dianae were studied in an open-top chamber experiment. Ozone was found to induce reductions in CO 2 assimilation and water use efficiency in the leaves of this species. These reductions were mainly related to a decline in the in vivo CO 2 fixation capacity of Rubisco (V c,max), rather than to stomatal limitations or photoinhibitory damage (F v:F m). In addition to chloroplast degeneration, other observed effects were callose accumulation, formation of pectinaceous wart-like cell wall exudates and phloem alterations. Moreover, ozone exposure significantly reduced root dry biomass. The possible relevance of these adverse effects for Mediterranean forbs is commented. These results show that endemic plants can be very sensitive to ozone, suggesting that risks associated with this pollutant should be taken into account for conservation purposes. © 2011 Elsevier Inc., We thank the Conselleria de Medi Ambient, Aigua, territory i Habitatge and FEDER funds (project VegetPollOzone, Interreg IIIb, Meddoc) for supporting parts of this study. The Instituto Universitario CEAM-UMH is partly supported by Generalitat Valenciana, Fundacion Bancaja, and benefits form the projects CONSOLIDER-INGENIO 2010 (GRACCIE) and Prometeo Program (Generalitat Valenciana). Emilio Laguna and Mari Carmen Escriva are also thanked for providing us the plants and, together with Joan Perez, for their comments on aspects of the biology of this species. Carmen Martin is thanked for taking care of the plants. Four anonymous referees provided very useful comments for improving the manuscript.

11 páginas, 2 tablas, 3 figuras., Within a European biomonitoring programme, Italian ryegrass (Lolium multiflorum Lam.) was employed as accumulative bioindicator of airbornetraceelements (As, Cd, Cr, Cu, Fe, Ni, Pb, Sb, V, Zn) in urban agglomerations. Applying a highly standardised method, grass cultures were exposed for consecutive periods of four weeks each to ambient air at up to 100 sites in 11cities during 2000–2002. Results of the 2001 exposure experiments revealed a clear differentiation of traceelementpollution within and among local monitoring networks. Pollution was influenced particularly by traffic emissions. Especially Sb, Pb, Cr, Fe, and Cu exhibited a very uneven distribution within the municipal areas with strong accumulation in plants from traffic-exposed sites in the city centres and close to major roads, and moderate to low levels in plants exposed at suburban or rural sites. Accumulation of Ni and V was influenced by other emission sources. The biomonitoring sites located in Spanish city centres featured a much higher pollution load by traceelements than those in other cities of the network, confirming previously reported findings obtained by chemical analyses of dust deposition and aerosols. At some heavily-trafficked sites, legal thresholds for Cu, Pb, and V contents in foodstuff and animal feed were reached or even surpassed. The study confirmed that the standardised grass exposure is a useful and reliable tool to monitor and to assess environmental levels of potentially toxic compounds of particulate matter., This study was supported by the LIFE Environment Programme of the European Commission under the grant LIFE/99/ENV/D/000453.CEAM is partly supported by Generalitat Valenciana, Bancaja, and CONSOLIDER-INGENIO 2010 (GRACCIE).

Ozone-like visible injury was detected on Hibiscus syriacus plants used as ornamental hedges. Weekly spray of the antiozonant ethylenediurea (EDU, 300 ppm) confirmed that the injury was induced by ambient ozone. EDU induced a 75% reduction in visible injury. Injury was more severe on the western than on the eastern exposure of the hedge. This factor of variability should be considered in ozone biomonitoring programmes. Seeds were collected and seedlings were artificially exposed to ozone in filtered vs. not-filtered (+30 ppb) Open-Top Chambers. The level of exposure inducing visible injury in the OTC seedlings was lower than that in the ambient-grown hedge. The occurrence of visible injury in the OTC confirmed that the ozone sensitivity was heritable and suggested that symptomatic plants of this deciduous shrub population can be successfully used as ozone bioindicators. EDU is recommended as a simple tool for diagnosing ambient ozone visible injury on field vegetation.

In general, it is difficult to measure air pollutant concentrations in remote areas, as they are mostly national parks and protected areas. Passive samplers provide an accurate and inexpensive method for measuring cumulative exposures of different air pollutants. They have been used to collect ozone data in both laboratory and field at different geographical scales. The objective of the present study is to fill the knowledge gap regarding air quality in remote areas of Spain, such as national parks and protected areas. Because there were no systematic data sets on the main air pollutants that could affect these areas, an air quality measurement network was established between 2001 and 2004 on 19 locations inside Spanish national parks and protected areas. The data collected suggest that ozone levels in mountainous areas are high enough to affect sensitive vegetation. Most of the locations registered moderate-to-high ozone levels, with important interannual variability. Altitudinal ozone gradients were observed in most of the parks with complex topography due to the establishment of local circulations that incorporate polluted air masses from polluted airsheds or even long-range transport (i.e., Canary Islands). Different latitude-dependent, yearly cycles were also observed, showing two, one, or no clear peaks depending on the region. These findings extend to the most southerly locations, except in the Canary Islands, where pollution transported from other regions in the upper transport layers probably led to the high concentrations observed.

International audience; Urban atmospheres contain complex mixtures of air pollutants including mutagenic and carcinogenic substances such as benzene, diesel soot, heavy metals and polycyclic aromatic hydrocarbons. In the frame of a European network for the assessment of air quality by the use of bioindicator plants, the Tradescantia micronucleus (Trad-MCN) test was applied to examine the genotoxicity of urban air pollution. Cuttings of Tradescantia clone #4430 were exposed to ambient air at 65 monitoring sites in 10 conurbations employing a standardised methodology. The tests revealed an elevated genotoxic potential mainly at those urban sites which were exposed to severe car traffic emissions. This bioassay proved to be a suitable tool to detect local ‘hot spots’ of mutagenic air pollution in urban areas. For its use in routine monitoring programmes, however, further standardisation of cultivation and exposure techniques is recommended in order to reduce the variability of results due to varying environmental conditions.

Ambient ozone concentrations were measured with passive samplers in the framework of the EU and UN/ECE Level II forest monitoring programme. Data from France, Italy, Luxembourg, Spain and Switzerland are reported for 2000-2002, covering the period from April to September. The number of plots increased from 67 in 2000 to 83 in 2002. The year 2001 experienced the highest ozone concentrations, reflecting more stable summer meteorological conditions. Average 6-month ozone concentrations above 45 ppb were measured this year in 40.3% of the plots, in contrast with the less than 21% measured in the other 2 years. Gradients of increasing ozone levels were observed from North to South and with altitude. Comments are made on the regional trends and on the time frame of the higher ozone episodes. Also, some recommendations enabling a better comparison between plots are provided.

Monitoring of forest condition and ozone (O3) at 83 sites in France, Italy, Luxenbourg, Spain and Switzerland resulted in a number of findings in relation to the knowledge of O3 exposure (concentration and cumulative AOT40), feasibility of the assessment of stomatal O3 flux and relationships between O3 and crown defoliation of beech and visible symptoms on native vegetation. However, the project provides evidence of issues to be addressed within the current monitoring system (data quality, validation sites and response indicators) and indications as to how the monitoring of O3 risk in the context of an effect-oriented monitoring program can be improved.

The onset and development of symptoms of three Sambucus species, S. ebulus, S. nigra and S. racemosa were studied in 2002 under three different experimental conditions, in charcoal filtered air (CF), and in two ozone enriched treatment: non filtered air plus 40 ppb ozone (NF+), and non filtered air plus 70 ppb ozone (NF++). The herb S. ebulus was more sensitive than the shrubs S. racemosa and S. nigra. Some plants of the three species showed visible injury below the AOT40 threshold of 10,000 ppb·h, established for protection of vegetation. Ozone produced a decrease in chlorophyll content in S. ebulus, and impaired both stomatal conductance and net photosynthesis in S. ebulus and S. nigra. A complementary study in 2004 with S. ebulus, confirmed a decrease in chlorophyll content after fumigation, associated to a decrease in N content of the leaves. Since S. ebulus is a widespread species in Europe and it is very sensitive to ozone, it could be a very appropriate plant for the biomonitoring studies across large areas in this continent.

EuroBionet, the ‘European Network for the Assessment of Air Quality by the Use of Bioindicator Plants’, is an EU-funded cooperative project currently consisting of public authorities and scientific institutes from 12 cities in 8 countries. In 2000, the bioindicator plants tobacco (Nicotiana tabacum Bel W3), poplar (Populus nigra ‘Brandaris’), spiderwort (Tradescantia sp. clone 4430), Italian rye grass (Lolium multiflorum italicum) and curly kale (Brassica oleracea acephala) were exposed to ambient air at 90 monitoring sites according to standardised methods. Visible injuries and growth parameters were assessed and the accumulation of toxic substances in leaves determined. The exposure of tobacco resulted in a gradient with low levels of ozone-induced foliar injury in N and NW Europe, and medium to high values in the southern and central regions. The results of heavy metal and sulphur analyses in rye grass samples generally showed low to very low sulphur and low to medium heavy metal concentrations in leaves. In some cities, however, local hot spots of heavy metal contamination were detected. Analyses of the PAH contents in curly kale leaves gave low to medium values, with locally elevated levels at traffic-exposed sites.