Ozone risk assessment with free-air controlled exposure (FACE) experiments: A critical revisit.

Since risk assessments of tropospheric ozone (O3) are crucial for agricultural and forestry sectors, there is a growing body for realistic assessments by a stomatal flux-based approach in Free-Air Controlled Exposure (FACE) facilities. Ozone risks are normally described as relative risks (RRs), which are calculated by assuming the biomass or yield at zero O3 dose as "reference". However, the estimation of the reference biomass or yield is challenging due to a lack of O3-clean-air treatment at the FACEs and the extrapolation without data in a low O3 range increases the bias for estimating the reference values. Here, we reviewed a current methodology for the risk assessment at FACEs and presented a simple and effective way ("modified Paoletti's approach") of defining RRs just using biomass or yield data with a range of expected impacts under the FACE conditions hypothesizing three possible scenarios based on prediction limits using 95% credible intervals (CI) (1. Best fit using the intercept as reference, 2. Optimistic scenario using a lower CI and 3. Worst scenario using an upper CI). As a result, O3-sensitive species show a relatively narrow effect range (optimistic vs. worst scenario) whereas a wide range of response may be possibly taken in resistant species. Showing a possible effect range allows for a comprehensive understanding of the potential risks and its uncertainties related to a species sensitivity to O3. As a supporting approach, we also recommend to use scientifically relevant tools (i.e., ethylenediurea treatments; mechanistic plant models) for strengthening the obtained results for the RRs against O3. Interestingly, the moderately sensitive or resistant species showed non-linear rather than linear dose-response relationships, suggesting a need for the flexible functional form for the risk assessment to properly describe the complex plant response such as hormesis, which depends on their plasticity to O3 stress.

Meteorological, chemical and biological evaluation of the coupled chemistry-climate WRF-Chem model from regional to urban scale. An impact-oriented application for human health.

Extreme climatic conditions, like heat waves or cold spells, associated to high concentrations of air pollutants are responsible for a broad range of effects on human health. Consequently, in the recent years, the question on how urban and peri-urban forests may improve both air quality and surface climate conditions at city-scale is receiving growing attention by scientists and policymakers, with previous studies demonstrating how nature-based solutions (NBS) may contribute to reduce the risk of population to be exposed to high pollutant levels and heat stress, preventing, thus, premature mortality. In this study we present a new modeling framework designed to simulate air quality and meteorological conditions from regional to urban scale, allowing thus to assess the impacts of both air pollution and heat stress on human health at urban level. To assess the model reliability, we evaluated the model's performances in reproducing several relevant meteorological, chemical, and biological variables. Results show how our modeling system can reliably reproduce the main meteorological, chemical, and biological variables over our study areas, thus this tool can be used to estimate the impact of air pollution and heat stress on human health. As an example of application, we show how common heat stress and air pollutant indices used for human health protection change when computed from regional to urban scale for the cities of Florence (Italy) and Aix en Provence (France).

Detection of morphological and eco-physiological traits of ornamental woody species to assess their potential Net O3 uptake.

Urban greening can improve cities' air quality by filtering the main gaseous pollutants such as tropospheric ozone (O3). However, the pollutant removal capacity offered by woody species strongly depends on eco-physiological and morphological traits. Woody species with higher stomatal conductance (gs) can remove more gases from the atmosphere, but other species can worsen air quality due to high O3 forming potential (OFP), based on their emitting rates of biogenic volatile organic compounds (bVOCs) and Leaf Mass per Area (LMA). Presently, there is a lack of data on eco-physiological (gs, bVOCs emissions) and foliar traits (LMA) for several ornamental species used in urban greening programs, which does not allow assessment of their O3 removal capacity and OFP. This study aimed to (i) parameterize gs, assess bVOCs emissions and LMA of 14 ornamental woody species commonly used in Mediterranean urban greening, and (ii) model their Net O3 uptake. The gs Jarvis model was parameterized considering various environmental conditions alongside isoprene and monoterpene foliar bVOCs emission rates trapped in the field and quantified by gas chromatography-mass spectrometry. The results are helpful for urban planning and landscaping; suggesting that Catalpa bignonioides and Gleditsia triacanthos have excellent O3 removal capacity due to their high maximum gs (gmax) equal to 0.657 and 0.597 mol H2O m-2 s-1. Regarding bVOCs, high isoprene (16.75 µg gdw-1 h-1) and monoterpene (13.12 µg gdw-1 h-1) emission rates were found for Rhamnus alaternus and Cornus mas. In contrast, no bVOCs emissions were detected for Camellia sasanqua and Paulownia tomentosa. In conclusion, 11 species showed a positive Net O3 uptake, while the use of large numbers of R. alaternus, C. mas, and Chamaerops humilis for urban afforestation planning are not recommended due to their potential to induce a deterioration of outdoor air quality.

Surface ozone risk to human health and vegetation in tropical region: The case of Thailand.

Tropospheric ozone (O3) is a threat to vegetation and human health over the world, in particular in Asia. Knowledge on O3 impacts on tropical ecosystems is still very limited. An O3 risk assessment to crops, forests, and people from 25 monitoring stations across the tropical and subtropical Thailand during 2005-2018 showed that 44% of sites exceeded the critical levels (CLs) of SOMO35 (i.e., the annual Sum Of daily maximum 8-h Means Over 35 ppb) for human health protection. The concentration-based AOT40 CL (i.e., sum of the hourly exceedances above 40 ppb for daylight hours during the assumed growing season) was exceeded at 52% and 48% of the sites where the main crops rice and maize are present, respectively, and at 88% and 12% of the sites where evergreen or deciduous forests are present, respectively. The flux-based metric PODY (i.e., Phytotoxic Ozone Dose above a threshold Y of uptake) was calculated and was found to exceed the CLs at 1.0%, 1.5%, 20.0%, 1.5%, 0% and 68.0% of the sites where early rice, late rice, early maize, late maize, evergreen forests, and deciduous forests can grow, respectively. Trend analysis indicated that AOT40 increased over the study period (+5.9% year-1), while POD1 decreased (- 5.3% year-1), suggesting that the role of climate change in affecting the environmental factors that control stomatal uptake cannot be neglected. These results contribute novel knowledge on O3 threat to human health, forest productivity, and food security in tropical and subtropical areas.

Strategic roadmap to assess forest vulnerability under air pollution and climate change.

Although it is an integral part of global change, most of the research addressing the effects of climate change on forests have overlooked the role of environmental pollution. Similarly, most studies investigating the effects of air pollutants on forests have generally neglected the impacts of climate change. We review the current knowledge on combined air pollution and climate change effects on global forest ecosystems and identify several key research priorities as a roadmap for the future. Specifically, we recommend (1) the establishment of much denser array of monitoring sites, particularly in the South Hemisphere; (2) further integration of ground and satellite monitoring; (3) generation of flux-based standards and critical levels taking into account the sensitivity of dominant forest tree species; (4) long-term monitoring of N, S, P cycles and base cations deposition together at global scale; (5) intensification of experimental studies, addressing the combined effects of different abiotic factors on forests by assuring a better representation of taxonomic and functional diversity across the ~73,000 tree species on Earth; (6) more experimental focus on phenomics and genomics; (7) improved knowledge on key processes regulating the dynamics of radionuclides in forest systems; and (8) development of models integrating air pollution and climate change data from long-term monitoring programs.

Species-specific variation of photosynthesis and mesophyll conductance to ozone and drought in three Mediterranean oaks.

Mesophyll conductance (gmCO2 ) is one of the most important components in plant photosynthesis. Tropospheric ozone (O3 ) and drought impair physiological processes, causing damage to photosynthetic systems. However, the combined effects of O3 and drought on gmCO2 are still largely unclear. We investigated leaf gas exchange during mid-summer in three Mediterranean oaks exposed to O3 (ambient [35.2 nmol mol-1 as daily mean]; 1.4 × ambient) and water treatments (WW [well-watered] and WD [water-deficit]). We also examined if leaf traits (leaf mass per area [LMA], foliar abscisic acid concentration [ABA]) could influence the diffusion of CO2 inside a leaf. The combination of O3 and WD significantly decreased net photosynthetic rate (PN ) regardless of the species. The reduction of photosynthesis was associated with a decrease in gmCO2 and stomatal conductance (gsCO2 ) in evergreen Quercus ilex, while the two deciduous oaks (Q. pubescens, Q. robur) also showed a reduction of the maximum rate of carboxylation (Vcmax ) and maximum electron transport rate (Jmax ) with decreased diffusive conductance parameters. The reduction of gmCO2 was correlated with increased [ABA] in the three oaks, whereas there was a negative correlation between gmCO2 with LMA in Q. pubescens. Interestingly, two deciduous oaks showed a weak or no significant correlation between gsCO2 and ABA under high O3 and WD due to impaired stomatal physiological behaviour, indicating that the reduction of PN was related to gmCO2 rather than gsCO2 . The results suggest that gmCO2 plays an important role in plant carbon gain under concurrent increases in the severity of drought and O3 pollution.

Experimental assessment of ozone risk on ecotypes of the tropical tree Moringa oleifera.

Ozone (O3) is an oxidative air pollutant that affects plant growth. Moringa oleifera is a tree species distributed in the tropical and subtropical regions. This species presents high morphological plasticity, which increases its ability to tolerate stressful conditions, but with no O3 risk assessment calculated so far. The present study assessed the O3 risk to different M. oleifera ecotypes using exposure-based index (AOT40) or flux-based index (PODy - where y is a threshold of O3 uptake). PODy considers the O3 uptake through the stomata and the consequence of environmental climate conditions on stomatal conductance (gsto); thus, it is efficient in assessing O3 risk. Five M. oleifera ecotypes were subjected to ambient (Amb.); middle (Mid. X1.5), and High (x2.0) O3 concentrations for 77 days in a free-air controlled exposure facility (FACE). Leaf biomass (LB) was evaluated, and the biomass loss was projected assuming a clean atmosphere (10 ppb as 24 h O3 average). The gsto parameterization was calculated using the Jarvis-type multiplicative algorithm considering several climate factors, i.e., light intensity, air temperature, air vapor pressure deficit, and AOT40. Ozone exposure harmed the LB of all ecotypes. The high gsto (~559 mmol H2O m-2 s-1) can be considered the reason for the species' O3 sensitivity. M. oleifera is adapted to hot climate conditions, and gsto was restricted with air temperature (Tmin) below ~ 9 °C. As expected, the PODy index performed better than the AOT40 for estimating the O3 effect on biomass losses. We recommend a y threshold of 4 nmol m-2 s-1 to incorporate O3 effects on M. oleifera LB. To not exceed a 4% reduction of LB for any M. oleifera genotype, we recommend the critical levels of 1.1 mmol m-2 POD4.

Stress markers and physiochemical responses of the Mediterranean shrub Phillyrea angustifolia under current and future drought and ozone scenarios.

Mediterranean plants are particularly threatened by the exacerbation of prolonged periods of summer drought and increasing concentrations of ground-level ozone (O3). The aims of the present study were to (i) test if selected markers (i.e., reactive oxygen species, ROS; malondialdehyde, MDA; photosynthetic pigments) are able to discriminate the oxidative pressure due to single and combined stress conditions, and (ii) elucidate the physiochemical adjustments adopted by Phillyrea angustifolia (evergreen woody species representative of the maquis, also known as narrow-leaved mock privet) to perceive and counter to drought and/or O3. Plants were grown from May to October under the combination of two levels of water irrigation [i.e., well-watered (WW) and water-stressed (WS)] and three levels of O3 [i.e., 1.0, 1.5 and 2.0 times the ambient air concentrations, i.e. AA (current O3 scenario), 1.5 × AA and 2.0 × AA (future O3 scenarios), respectively], using a new-generation O3 Free Air Controlled Exposure (FACE) system. Overall, this species appeared relatively sensitive to drought (e.g., net CO2 assimilation rate and stomatal conductance significantly decreased, as well as total chlorophyll and carotenoid contents), and tolerant to O3 (e.g., as confirmed by the absence of visible foliar injury, the unchanged values of total carotenoids, and the detrimental effects on stomatal conductance, total chlorophylls and terpene emission only under elevated O3 concentrations). The combination of both stressors led to harsher oxidative stress. Only when evaluated together (i.e., combining the information provided by the analysis of each stress marker), ROS, MDA and photosynthetic pigments, were suitable stress markers to discriminate the differential oxidative stress induced by drought and increasing O3 concentrations applied singly or in combination: (i) all these stress markers were affected under drought per se; (ii) hydrogen peroxide (H2O2) and MDA increased under O3per se, following the gradient of O3 concentrations (H2O2: about 2- and 4-fold higher; MDA: +22 and + 91%; in 1.5 × AA_WW and 2.0 × AA_WW, respectively); (iii) joining together the ROS it was possible to report harsher effects under 2.0 × AA_WS and 1.5 × AA_WS (both anion superoxide and H2O2 increased) than under 2.0 × AA_WW (only H2O2 increased); and (iv) MDA showed harsher effects under 2.0 × AA_WS than under 1.5 × AA_WS (increased by 49 and 18%, respectively). Plants activated physiological and biochemical adjustments in order to partially avoid (e.g., stomatal closure) and tolerate (e.g., increased terpene emission) the effects of drought when combined with increasing O3 concentrations, suggesting that the water use strategy (isohydric) and the sclerophyllous habit can further increase the plant tolerance to environmental constraints in the Mediterranean area.

Date palm responses to a chronic, realistic ozone exposure in a FACE experiment.

Date palms are highly economically important species in hot arid regions, which may suffer ozone (O3) pollution equivalently to heat and water stress. However, little is known about date palm sensitivity to O3. Therefore, to identify their resistance mechanisms against elevated O3, physiological parameters (leaf gas exchange, chlorophyll fluorescence and leaf pigments) and biomass growth responses to realistic O3 exposure were tested in an isoprene-emitting date palm (Phoenix dactylifera L. cv. Nabut Saif) by a Free-Air Controlled Exposure (FACE) facility with three levels of O3 (ambient [AA, 45 ppb as 24-h average], 1.5 x AA and 2 x AA). We found a reduction of photosynthesis only at 2 x AA although some foliar traits known as early indicators of O3 stress responded already at 1.5 x AA, such as increased dark respiration, reduced leaf pigment content, reduced maximum quantum yield of PSII, inactivation of the oxygen evolving complex of PSII and reduced performance index PITOT. As a result, O3 did not affect most of the growth parameters although significant declines of root biomass occurred only at 2 x AA. The major mechanism in date palm for reducing the severity of O3 impacts was a restriction of stomatal O3 uptake due to low stomatal conductance and O3-induced stomatal closure. In addition, an increased respiration in elevated O3 may indicate an enhanced capacity of catabolizing metabolites for detoxification and repair. Interestingly, date palm produced low amounts of monoterpenes, whose emission was stimulated in 2 x AA, although isoprene emission declined at both 1.5 and 2 x AA. Our results warrant more research on a biological significance of terpenoids in plant resistance against O3 stress.

Water use strategy affects avoidance of ozone stress by stomatal closure in Mediterranean trees-A modelling analysis.

Both ozone (O3 ) and drought can limit carbon fixation by forest trees. To cope with drought stress, plants have isohydric or anisohydric water use strategies. Ozone enters plant tissues through stomata. Therefore, stomatal closure can be interpreted as avoidance to O3 stress. Here, we applied an optimization model of stomata involving water, CO2 , and O3 flux to test whether isohydric and anisohydric strategies may affect avoidance of O3 stress by stomatal closure in four Mediterranean tree species during drought. The data suggest that stomatal closure represents a response to avoid damage to the photosynthetic mechanisms under elevated O3 depending on plant water use strategy. Under high-O3 and well-watered conditions, isohydric species limited O3 fluxes by stomatal closure, whereas anisohydric species activated a tolerance response and did not actively close stomata. Under both O3 and drought stress, however, anisohydric species enhanced the capacity of avoidance by closing stomata to cope with the severe oxidative stress. In the late growing season, regardless of the water use strategy, the efficiency of O3 stress avoidance decreased with leaf ageing. As a result, carbon assimilation rate was decreased by O3 while stomata did not close enough to limit transpirational water losses.

Interactive effect of leaf age and ozone on mesophyll conductance in Siebold's beech.

Mesophyll conductance (Gm ) is one of the most important factors determining photosynthesis. Tropospheric ozone (O3 ) is known to accelerate leaf senescence and causes a decline of photosynthetic activity in leaves. However, the effects of age-related variation of O3 on Gm have not been well investigated, and we, therefore, analysed leaf gas exchange data in a free-air O3 exposure experiment on Siebold's beech with two levels (ambient and elevated O3 : 28 and 62 nmol mol-1 as daylight average, respectively). In addition, we examined whether O3 -induced changes on leaf morphology (leaf mass per area, leaf density and leaf thickness) may affect CO2 diffusion inside leaves. We found that O3 damaged the photosynthetic biochemistry progressively during the growing season. The Gm was associated with a reduced photosynthesis in O3 -fumigated Siebold's beech in August. The O3 -induced reduction of Gm was negatively correlated with leaf density, which was increased by elevated O3 , suggesting that the reduction of Gm was accompanied by changes in the physical structure of mesophyll cells. On the other hand, in October, the O3 -induced decrease of Gm was diminished because Gm decreased due to leaf senescence regardless of O3 treatment. The reduction of photosynthesis in senescent leaves after O3 exposure was mainly due to a decrease of maximum carboxylation rate (Vcmax ) and/or maximum electron transport rate (Jmax ) rather than diffusive limitations to CO2 transport such as Gm . A leaf age×O3 interaction of photosynthetic response will be a key for modelling photosynthesis in O3 -polluted environments.

Hyperspectral Reflectance of Light-Adapted Leaves Can Predict Both Dark- and Light-Adapted Chl Fluorescence Parameters, and the Effects of Chronic Ozone Exposure on Date Palm (Phoenix dactylifera).

High-throughput and large-scale measurements of chlorophyll a fluorescence (ChlF) are of great interest to investigate the photosynthetic performance of plants in the field. Here, we tested the capability to rapidly, precisely, and simultaneously estimate the number of pulse-amplitude-modulation ChlF parameters commonly calculated from both dark- and light-adapted leaves (an operation which usually takes tens of minutes) from the reflectance of hyperspectral data collected on light-adapted leaves of date palm seedlings chronically exposed in a FACE facility to three ozone (O3) concentrations (ambient air, AA; target 1.5 × AA O3, named as moderate O3, MO; target 2 × AA O3, named as elevated O3, EO) for 75 consecutive days. Leaf spectral measurements were paired with reference measurements of ChlF, and predictive spectral models were constructed using partial least squares regression. Most of the ChlF parameters were well predicted by spectroscopic models (average model goodness-of-fit for validation, R2: 0.53-0.82). Furthermore, comparing the full-range spectral profiles (i.e., 400-2400 nm), it was possible to distinguish with high accuracy (81% of success) plants exposed to the different O3 concentrations, especially those exposed to EO from those exposed to MO and AA. This was possible even in the absence of visible foliar injury and using a moderately O3-susceptible species like the date palm. The latter view is confirmed by the few variations of the ChlF parameters, that occurred only under EO. The results of the current study could be applied in several scientific fields, such as precision agriculture and plant phenotyping.

Isoprene is more affected by climate drivers than monoterpenes: A meta-analytic review on plant isoprenoid emissions.

Isoprene and monoterpenes (MTs) are among the most abundant and reactive volatile organic compounds produced by plants (biogenic volatile organic compounds). We conducted a meta-analysis to quantify the mean effect of environmental factors associated to climate change (warming, drought, elevated CO2 , and O3 ) on the emission of isoprene and MTs. Results indicated that all single factors except warming inhibited isoprene emission. When subsets of data collected in experiments run under similar change of a given environmental factor were compared, isoprene and photosynthesis responded negatively to elevated O3 (-8% and -10%, respectively) and drought (-15% and -42%), and in opposite ways to elevated CO2 (-23% and +55%) and warming (+53% and -23%, respectively). Effects on MTs emission were usually not significant, with the exceptions of a significant stimulation caused by warming (+39%) and by elevated O3 (limited to O3 -insensitive plants, and evergreen species with storage organs). Our results clearly highlight individual effects of environmental factors on isoprene and MT emissions, and an overall uncoupling between these secondary metabolites produced by the same methylerythritol 4-phosphate pathway. Future results from manipulative experiments and long-term observations may help untangling the interactive effects of these factors and filling gaps featured in the current meta-analysis.

A quantitative assessment of hormetic responses of plants to ozone.

Evaluations of ozone effects on vegetation across the globe over the last seven decades have mostly incorporated exposure levels that were multi-fold the preindustrial concentrations. As such, global risk assessments and derivation of critical levels for protecting plants and food supplies were based on extrapolation from high to low exposure levels. These were developed in an era when it was thought that stress biology is framed around a linear dose-response. However, it has recently emerged that stress biology commonly displays non-linear, hormetic processes. The current biological understanding highlights that the strategy of extrapolating from high to low exposure levels may lead to biased estimates. Here, we analyzed a diverse sample of published empirical data of approximately 500 stimulatory, hormetic-like dose-responses induced by ozone in plants. The median value of the maximum stimulatory responses induced by elevated ozone was 124%, and commonly <150%, of the background response (control), independently of species and response variable. The maximum stimulatory response to ozone was similar among types of response variables and major plant species. It was also similar among clades, between herbaceous and woody plants, between deciduous and evergreen trees, and between annual and perennial herbaceous plants. There were modest differences in the stimulatory response between genera and between families which may reflect different experimental designs and conditions among studies. The responses varied significantly upon type of exposure system, with open-top chambers (OTCs) underestimating the maximum stimulatory response compared to free-air ozone-concentration enrichment (FACE) systems. These findings suggest that plants show a generalized hormetic stimulation by ozone which is constrained within certain limits of biological plasticity, being highly generalizable, evolutionarily based, and maintained over ecological scales. They further highlight that non-linear responses should be taken into account when assessing the ozone effects on plants.

Effects of nitrogen and phosphorus imbalance on photosynthetic traits of poplar Oxford clone under ozone pollution.

Ozone (O3) pollution and the availability of nitrogen (N) and phosphorus (P) in the soil both affect plant photosynthesis and chlorophyll (Chl) content, but the interaction of O3 and nutrition is unclear. We postulated that the nutritional condition changes plant photosynthetic responses to O3. An O3-sensitive poplar clone (Oxford) was subject to two N levels (N0, 0 kg N ha- 1; N80, 80 kg N ha- 1), two P levels (P0, 0 kg P ha- 1; P80, 80 kg P ha- 1) and three levels of O3 exposure (ambient concentration, AA; 1.5 × AA; 2.0 × AA) over a growing season in an O3 free air controlled exposure (FACE) facility. The daily change of leaf gas exchange and dark respiration (Rd) were investigated at mid-summer (August). Chl a fluorescence was measured three times in July, August and September. At the end of the growing season, Chl content was measured. It was found that Chl content, the maximum quantum yield (Fv/Fm), Chl a fluorescence performance index (PI) and gas exchange were negatively affected by elevated O3. Phosphorus may mitigate the O3-induced reduction of the ratio of photosynthesis to stomatal conductance, while it exacerbated the O3-induced loss of Fv/Fm. Nitrogen alleviated negative effects of O3 on Fv/Fm and PI in July. Ozone-induced loss of net photosynthetic rate was mitigated by N in medium O3 exposure (1.5 × AA). However, such a mitigation effect was not observed in the higher O3 level (2.0 × AA). Nitrogen addition exacerbated O3-induced increase of Rd suggesting an increased respiratory carbon loss in the presence of O3 and N. This may result in a further reduction of the net carbon gain for poplars exposed to O3.

Protecting the photosynthetic performance of snap bean under free air ozone exposure.

Tropospheric ozone (O3) is a major air pollutant and causes serious injury to vegetation. To protect sensitive plants from O3 damage, several agrochemicals have been assessed, including cytokinin (e.g., kinetin, KIN) and ethylenediurea (EDU) with cytokinin-like activity. In higher plant, leaves are primarily injured by O3 and protective agrochemicals are often applied by leaf spraying. To our knowledge, the mitigating abilities of EDU and KIN have not been compared directly in a realistic setup. In the present research, impacts of elevated O3 (2× ambient O3, 24hr per day, for 8days) on an O3 sensitive line (S156) of snap bean (Phaseolus vulgaris), which is often used for biomonitoring O3 pollution, were studied in a free air controlled exposure system. The day before starting the O3 exposure, plants were sprayed with a solution of EDU (300ppm), KIN (1mmol/L) or distilled water, to compare their protective abilities. The results demonstrated that 2× ambient O3 inhibited net photosynthetic rate and stomatal conductance, increased the minimal fluorescence yield of the dark-adapted state, decreased the maximal quantum yield of PSII photochemistry, and led to visible injury. KIN and EDU alleviated the reduction of the photosynthetic performance, and visible injury under O3 fumigation. The plants sprayed with EDU showed greater ability to mitigate the O3 damage than those sprayed with KIN. Chlorophyll fluorescence imaging may have detected more precisely the differences in O3 response across the leaf than the conventional fluorometer.

Photosynthetic responses of Monarch birch seedlings to differing timings of free air ozone fumigation.

To study the effects of different periods of ozone (O3) fumigation on photosynthesis in leaves of the Monarch birch (Betula maximowicziana), we undertook free air O3 fumigation to Monarch birch seedlings at a concentration of 60 nmol mol(-1) during daytime. Plants were exposed to O3 at early, late or both periods in the growing season. The light-saturated net photosynthetic rate (A(sat)) in July and August was reduced by O3 exposure through a reduction in the maximum rate of carboxylation (V(c,max)). In early September, on the other hand, despite a reduction in V(c,max), A(sat) was not reduced by O3 due to a counteracting increase in the stomatal conductance. Through the experiment, there was no difference in sensitivity to O3 between maturing and matured leaves. We analyzed the relationship between A(sat), V(c,max) and accumulated stomatal O3 flux (AF(st)). Whereas V(c,max) decreased with increasing AF(st), the correlation between A(sat) and AF(st) was weak because the response of stomatal conductance to O3 was affected by season. We conclude photosynthetic response of Monarch birch to O3 exposure changes with season. This is due to the inconstant stomatal response to O3 but not due to the respose of biochemical assimilation capacity in chloroplasts.

Allocation of Nutrients and Leaf Turnover Rate in Poplar under Ambient and Enriched Ozone Exposure and Soil Nutrient Manipulation.

An excess of ozone (O3) is currently stressing plant ecosystems and may negatively affect the nutrient use of plants. Plants may modify leaf turnover rates and nutrient allocation at the organ level to counteract O3 damage. We investigated leaf turnover rate and allocation of primary (C, N, P, K) and secondary macronutrients (Ca, S, Mg) under various O3 treatments (ambient concentration, AA, with a daily hourly average of 35 ppb; 1.5 × AA; 2.0 × AA) and fertilization levels (N: 0 and 80 kg N ha-1 y-1; P: 0 and 80 kg N ha-1 y-1) in an O3-sensitive poplar clone (Oxford: Populus maximowiczii Henry × P. berolinensis Dippel) in a Free-Air Controlled Exposure (FACE) experiment. The results indicated that both fertilization and O3 had a significant impact on the nutrient content. Specifically, fertilization and O3 increased foliar C and N contents (+5.8% and +34.2%, respectively) and root Ca and Mg contents (+46.3% and +70.2%, respectively). Plants are known to increase the content of certain elements to mitigate the damage caused by high levels of O3. The leaf turnover rate was accelerated as a result of increased O3 exposure, indicating that O3 plays a main role in influencing this physiological parameter. A PCA result showed that O3 fumigation affected the overall allocation of primary and secondary elements depending on the organ (leaves, stems, roots). As a conclusion, such different patterns of element allocation in plant leaves in response to elevated O3 levels can have significant ecological implications.

An assessment of ozone risk for date palm suggests that phytotoxic ozone dose nonlinearly affects carbon gain.

Tropospheric ozone (O3) is a significant phytotoxic air pollutant that has a negative impact on plant carbon gain. Although date palm (Phoenix dactylifera L.) is a globally important crop in arid or semi-arid regions, so far O3 risk assessment for this species has not been reported. This study estimated leaf- and plant-level photosynthetic CO2 uptake for understanding how elevated levels of O3 affects date palm biomass growth. Ozone risks to date palm plants were assessed based on exposure- (AOT40) or flux-based indices (Phytotoxic Ozone Dose, PODy, where y is a threshold of uptake). For this purpose, plants were exposed to three levels of O3 [ambient air, AA (45 ppb as daily average); 1.5 × AA; 2.0 × AA] for 92 days in an O3 Free-Air Controlled Exposure facility. According to the model simulations, the negative effects of O3 on plant-level net photosynthetic CO2 uptake were attributed to reduced gross photosynthetic carbon gain and increased respiratory carbon loss. Season-long O3 exposure and elevated temperatures promoted the negative O3 effect because of a further increase of respiratory carbon loss, which was caused by increased leaf temperature due to stomatal closure. POD1 nonlinearly affected the photosynthetic CO2 uptake, which was closely related to the variation of dry mass increment during the experiment. Although the dose-response relationship suggested that a low O3 dose (POD1 < 5.2 mmol m-2) may even positively affect photosynthetic CO2 uptake in date palms, stomatal O3 uptake at the current ambient O3 levels has potentially a negative impact on date palm growth. The results indicate 5.8 mmol m-2 POD1 or 21.1 ppm h AOT40 as critical levels corresponding to a 4% reduction of net CO2 uptake for date palm, suggesting that this species can be identified as a species moderately sensitive to O3.

Untangling the role of leaf age specific osmoprotectant and antioxidant responses of two poplar clones under increasing ozone concentrations.

Plants possess different degrees of tolerance to abiotic stress, which can mitigate the detrimental effect of environmental inputs affecting carbon balance. Less is known about the functions of osmoprotectants in scavenging of reactive oxygen species (ROS), generated at different sites depending on leaf age. This study aimed to clarify the osmotic adjustments adopted by old and young leaves of Oxford and I-214 poplar clones [differing in ozone (O3) sensitivity] to cope with three levels of O3 [ambient (AA), and two elevated O3 levels]. In both clones, the impact of intermediate O3 concentrations (1.5 × AA) on ROS production appeared to be leaf age-specific, given the accumulation of hydrogen peroxide (H2O2) observed only in old leaves of the Oxford plants and in young leaves of the I-214 ones (2- fold higher than AA and +79%, respectively). The induction of an oxidative burst was associated with membrane injury, indicating an inadequate response of the antioxidative systems [decrease of lutein and ß-carotene (-37 and -85% in the old leaves of the Oxford plants), accumulation of proline and tocopherols (+60 and +12% in the young leaves of the I-214 ones)]. Intermediate O3 concentrations reacted with unsaturated lipids of the plasma membrane in old and young leaves of the Oxford plants, leading to an increase of malondialdehyde by-products (more than 2- fold higher than AA), while no effect was recorded for I-214. The impact of the highest O3 concentrations (2.0 × AA) on ROS production did not appear clone-specific, which may react with cell wall components by leading to oxidative pressure. Outcomes demonstrated the ability of young leaves of I-214 plants in contain O3 phytotoxic effects.