Ozone risk assessment of common cypress (Cupressus sempervirens L.) clones and effects of Seiridium cardinale infection.

Cupressus sempervirens is a relevant species in the Mediterranean for its cultural, economic and landscape value. This species is threatened by Seiridium cardinale, the causal agent of the cypress canker disease (CCD). The effects of biotic stressors on O3 risk assessment are unknown and a comprehensive O3 risk assessment in C. sempervirens is missing. To fill these gaps, two clones of C. sempervirens, one resistant (Clone R) and one susceptible to CCD (Clone S), were subjected to three levels of O3 (Ambient Air - AA; 1.5 × AA; 2.0 × AA) for two consecutive years in an O3-free-air controlled exposure facility and artificially inoculated with S. cardinale. Both the exposure- (AOT40) and flux-based (PODy) indices were tested. We found that PODy performed better than AOT40 to assess O3 effects on biomass and the critical level for a 4% biomass loss was 2.51 mmol/m2 POD2. However, significant O3 dose-response relationships were not found for the inoculated cypresses because the combination of middle level O3 (1.5 × AA) and inoculation stimulated a biomass growth in Clone S as hormetic response. Moreover, we found a different inter-clonal response to both stressors with a statistically significant reduction of total and belowground biomass following O3, and lower root biomass in Clone S than in Clone R following pathogen infection. In summary, Clone R was more resistant to O3, and inoculation altered O3 risk via an hormetic effect on biomass. These results warrant further studies on how biotic stressors affect O3 responses and risk assessment.

Stress physiology of Moringa oleifera under tropospheric ozone enrichment: An ecotype-specific investigation into growth, nonstructural carbohydrates, and polyphenols.

Ozone (O3) is an oxidative pollutant that significantly threatens plant development and ecological dynamics. The present study explores the impact of O3 on Moringa (Moringa oleifera) ecotypes when exposed to ambient and elevated O3 levels. Elevated O3 concentrations resulted in significant reductions in total biomass for all ecotypes. Photosynthetic parameters, including stomatal conductance (gsto), CO2 assimilation (Pn), and carboxylation efficiency (K), decreased under elevated O3 in some ecotypes, indicating a detrimental effect on carbon assimilation. Nonstructural carbohydrate (NSC) levels in roots varied among ecotypes, with significant reductions in starch content observed under elevated O3, suggesting a potential shift towards soluble sugar accumulation and reallocation for antioxidant defense. Secondary metabolite analysis revealed increased polyphenol production, particularly quercetin derivatives, under elevated O3 in specific ecotypes, highlighting their role in mitigating oxidative stress. Interestingly, the glucosinolate content also varied, with some ecotypes exhibiting increased levels, suggesting a complex regulatory mechanism in response to O3 exposure. The study underscores the intrinsic variability among Moringa ecotypes in response to O3 stress, emphasizing the importance of genetic diversity for adaptation. The findings indicate that Moringa's metabolic plasticity, including shifts in NSC and SM production, plays a crucial role in its defense mechanisms against O3-induced oxidative stress. These insights are vital for optimizing the cultivation and utilization of Moringa in diverse environmental conditions, particularly in regions with elevated O3 levels.

Unraveling the difference of sensitivity to ozone between non-hybrid native poplar and hybrid poplar clones: A flux-based dose-response analysis.

Poplars are economically important tree crops and biologically important model plants, which are known to be sensitive to ozone (O3). Although surface O3 is considered as a significant global environmental issue because of its phytotoxicity and greenhouse effect, the knowledge of the dose-response (DR) relationships in poplars for the assessment of O3 risk is still limited. Hence, this study aimed at collecting data of studies with manipulative O3 exposures of poplars within FACE (Free Air Concentration Enhancement) and OTC (Open-Top Chamber) facilities. The datasets contain studies on hybrid poplar clones and a non-hybrid native poplar (Populus nigra L.) reporting both AOT40 (Accumulated exposure Over a Threshold of 40 ppb) and POD1 (Phytotoxic Ozone Dose above a threshold of 1 nmol m-2 Projected Leaf Area [PLA] s-1) to compare exposure- and flux-based indices. As a result, linear regression analysis showed that the flux-based POD1 was better than the exposure-based AOT40 to explain the biomass response of poplars to O3. From the DR relationships, a critical level (CL) of 5.7 mmol m-2 POD1 has been derived corresponding to 4% biomass growth reduction for hybrid poplar clones, which can be considered very sensitive to O3, while the non-hybrid native poplar was less sensitive to O3 (CL: 10.3 mmol m-2 POD1), although the potential risk of O3 for this taxon is still high due to very high stomatal conductance. Moreover, the different experimental settings (OTC vs. FACE) have affected the AOT40-based DR relationships but not the POD1-based DR relationships, suggesting that poplar responses to O3 were principally explained by stomatal O3 uptake regardless of the different experimental settings and exposure patterns. These results highlight the importance of the flux-based approach, especially when scaling up from experimental datasets to the O3 risk assessment for poplars at the regional or global scale.

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).

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.

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.

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.

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.

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.

The magpie and the grapes: increasing ozone exposure impacts fruit consumption by a common corvid in a suburban environment.

BACKGROUND: The Eurasian magpie Pica pica is a resident bird species able to colonize farmlands and anthropized environments. This corvid shows a wide trophic spectrum by including fruits, invertebrates, small vertebrates and carcasses in its diet. A camera-trap experiment was carried out to test the effect of different ozone (O3 ) concentrations on potted Vitis vinifera plants, which resulted in different grape consumption rates by suburban birds. The test was performed at an Ozone-Free Air Controlled Exposure (FACE) facility, consisting of nine plots with three ozone (O3 ) levels: AA (ambient O3 concentration); and two elevated O3 levels, 1.5× AA (ambient air with a 50% increase in O3 concentration) and 2.0× AA (ambient air with a 100% increase in O3 concentration). Camera-traps were located in front of each treatment area and kept active for 24 h day-1 and for 5 days at a time over a period of 3 months to monitor grape consumption by birds. RESULTS: We collected a total of 38 videos. Eurasian magpies were the only grape consumers, with a total of 6.7 ± 3.3 passages per hour (mean ± SD) and no differences across the different O3 treatments. Grapes in the AA treatment were consumed significantly more quickly than those in the 1.5× AA treatment, which in turn, were consumed faster than those in the 2.0× AA treatment. At 3 days from the start of treatment, 94%, 53% and 22% berries from the AA, 1.5× AA and 2.0× AA treatments had been eaten, respectively. When the O3 was turned off, berries were consumed at the same rate among treatments. CONCLUSION: Increasing O3 concentrations limited grape consumption by magpies probably because O3 acted as a deterrent for magpies, although the lower sugar content recorded in the 2.0× AA berries did not affect the consumption when O3 was turned off. Our results provided valuable insights to mitigate human-wildlife conflicts in suburban environments. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A three-year free-air experimental assessment of ozone risk on the perennial Vitis vinifera crop species.

Tropospheric ozone (O3) is an oxidative air pollutant that promotes damage to several crops, including grapevine, which is considered moderately resistant to O3 stress. To study the O3 effect on this perennial crop species under realistic environmental conditions, a three-year experiment was performed using an innovative O3-FACE facility located in the Mediterranean climate region, where the target species, Vitis vinifera cv. "Cabernet sauvignon", was exposed to three O3 levels: ambient (AA), 1.5 × ambient (×1.5), and 2 × ambient (×2.0). A stomatal conductance model parameterization was conducted, and O3-exposure (AOT40) and flux-based indices (PODy) were estimated. An assessment of O3-induced visible foliar injury (O3_VFI) was conducted by estimating VFI_Incidence (percentage of symptomatic leaves per branch) and VFI_Severity (average percentage of O3_VFI surface in symptomatic leaves). Biomass parameters were used to assess the cumulative O3 effect and calculate the most appropriate critical levels (CL) for a 5% yield loss and for the induction of 5, 10, and 15% of O3_VFI. We confirmed that the O3 effect on this grapevine variety VFI was cumulative and that POD0 values accumulated over the two or three years preceding the assessment were better related to the response variables than single-year values, with the response increasing with increasing O3 level. The estimated CL for 5% yield loss based on the O3-exposure index was 25 ppm h AOT40 and 21 or 23 ppm h for a 10% of VFI_Incidence or VFI_Severity, respectively. The suggested flux-based index value for 5% yield loss was 5.2 POD3 mmol m-2, and for 10% of VFI_Incidence or VFI_Severity, the values were 7.7 or 8.6 POD3 mmol m-2, respectively. The results presented in this study demonstrate that O3 risk assessment for this grapevine varietyproduces consistent and comparable results when using either yield or O3_VFI as response parameter.

Impacts of ground-level ozone on sugarcane production.

Sugarcane is a vital commodity crop often grown in (sub)tropical regions which have been experiencing a recent deterioration in air quality. Unlike for other commodity crops, the risk of air pollution, specifically ozone (O3), to this C4 crop has not yet been quantified. Yet, recent work has highlighted both the potential risks of O3 to C4 bioenergy crops, and the emergence of O3 exposure across the tropics as a vital factor determining global food security. Given the large extent, and planned expansion of sugarcane production in places like Brazil to meet global demand for biofuels, there is a pressing need to characterize the risk of O3 to the industry. In this study, we sought to a) derive sugarcane O3 dose-response functions across a range of realistic O3 exposure and b) model the implications of this across a globally important production area. We found a significant impact of O3 on biomass allocation (especially to leaves) and production across a range of sugarcane genotypes, including two commercially relevant varieties (e.g. CTC4, Q240). Using these data, we calculated dose-response functions for sugarcane and combined them with hourly O3 exposure across south-central Brazil derived from the UK Earth System Model (UKESM1) to simulate the current regional impact of O3 on sugarcane production using a dynamic global vegetation model (JULES vn 5.6). We found that between 5.6 % and 18.3 % of total crop productivity is likely lost across the region due to the direct impacts of current O3 exposure. However, impacts depended critically on the substantial differences in O3 susceptibility observed among sugarcane genotypes and how these were implemented in the model. Our work highlights not only the urgent need to fully elucidate the impacts of O3 in this important bioenergetic crop, but the potential implications air quality may have upon tropical food production more generally.

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.

Functional responses of two Mediterranean pine species in an ozone Free-Air Controlled Exposure (FACE) experiment.

Effects of the phytotoxic and widespread ozone (O3) pollution may be species specific, but knowledge on Mediterranean conifer responses to long-term realistic exposure is still limited. We examined responses regarding to photosynthesis, needle biochemical stress markers and carbon and nitrogen (N) isotopes of two Mediterranean pine species (Pinus halepensis Mill. and Pinus pinea L.). Seedlings were grown in a Free-Air Controlled Exposure experiment with three levels of O3 (ambient air, AA [38.7 p.p.b. as daily average]; 1.5 × AA and 2.0 × AA) during the growing season (May-October 2019). In P. halepensis, O3 caused a significant decrease in the photosynthetic rate, which was mainly due to a reduction of both stomatal and mesophyll diffusion conductance to CO2. Isotopic analyses indicated a cumulative or memory effect of O3 exposure on this species, as the negative effects were highlighted only in the late growing season in association with a reduced biochemical defense capacity. On the other hand, there was no clear effect of O3 on photosynthesis in P. pinea. However, this species showed enhanced N allocation to leaves to compensate for reduced photosynthetic N- use efficiency. We conclude that functional responses to O3 are different between the two species determining that P. halepensis with thin needles was relatively sensitive to O3, while P. pinea with thicker needles was more resistant due to a potentially low O3 load per unit mass of mesophyll cells, which may affect species-specific resilience in O3-polluted Mediterranean pine forests.

Ethylenediurea protects against ozone phytotoxicity not by adding nitrogen or controlling stomata in a stomata-unresponsive hybrid poplar.

Ozone (O3) pollution is a persistent environmental issue worldwide, which causes widespread damage to vegetation, deteriorating plant health and reducing plant productivity. Ethylenediurea (EDU) is a synthetic chemical that has been widely applied in scientific studies as a protectant against O3 phytotoxicities. Despite four decades of active research, the exact mechanisms to explain its mode of action remain unclear. Here, we aimed to reveal whether EDU's phytoprotective property is due to its control over stomatal regulation and/or its action as a nitrogen (N) fertilizer, utilizing stomatal-unresponsive plants of a hybrid poplar (Populus koreana × trichocarpa cv. Peace) grown in a free-air O3-concenctration enrichment (FACE) facility. Plants were treated with water (WAT), EDU (400 mg L-1), or EDU's constitutive amount of N every nine days, and exposed to ambient (AOZ) or elevated (EOZ) O3 during a growing season (June-September). EOZ led to extensive foliar injuries (but protected against rust disease), lower photosynthetic rate (A), impaired dynamics of responses of A to changes in light intensity, and smaller total plant leaf area. EDU protected against common phytotoxicities caused by EOZ without inducing stomatal closure, since stomatal conductance (gs) was generally unresponsive to the experimental treatments. EDU also modulated the dynamic response of A to light fluctuations under O3 stress. N addition acted as a fertilizer but did not satisfactorily protect plants against O3 phytotoxicities. The results suggest that EDU protects against O3 phytotoxicity not by adding N or controlling stomata, which provides a new insight into our understanding of the mode of action of EDU as a protectant against O3 phytotoxicity.

Chronic ozone exposure impairs the mineral nutrition of date palm (Phoenix dactylifera) seedlings.

Chronic ozone (O3) exposure in the atmosphere preferentially disturbs metabolic processes in the roots rather than the shoot as a consequence of reduced photosynthesis and carbohydrate allocation from the leaves to the roots. The aim of the present study was to elucidate if mineral nutrition is also impaired by chronic O3 exposure. For this purpose, date palm (Phoenix dactylifera) plants were fumigated with ambient, 1.5 × ambient and 2 × ambient O3 in a free air controlled exposure (FACE) system for one growing season and concentrations of major nutrients were analyzed in leaves and roots. In addition, concentrations of C and N and their partitioning between different metabolic C and N pools were determined in both organs. The results showed that calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn), sodium (Na) and potassium (K) acquisition by roots was diminished by O3 exposure of the shoot. For Ca, Mg, Fe and Zn reduced uptake by the roots was combined with reduced allocation to the shoot, resulting in a decline of foliar concentrations; for Na and K, allocation to the shoot was maintained at the expense of the roots. Thus, elevated O3 impaired both mineral uptake by the roots and partitioning of minerals between roots and shoots, but in an element specific way. Thereby, elevated O3 affected roots and shoots differently already after one growing season. However, considerable changes in total C and N concentrations and their partitioning between different metabolic pools upon chronic O3 exposure were not observed in either leaves or roots, except for reduced foliar lignin concentrations at 2 × ambient O3. Significant differences in these parameters were shown between leaves and roots independent of O3 application. The physiological consequences of the effects of chronic O3 exposure on mineral acquisition and partitioning between leaves and roots are discussed.

Visible Foliar Injury and Ecophysiological Responses to Ozone and Drought in Oak Seedlings.

To verify the responses of visible foliar injury (VFI), we exposed seedlings of three oak species for 4.5 months in an open air facility, using differing ozone (O3) and drought treatments: O3 (three levels from ambient to ×1.4 ambient), and drought (three levels of irrigation from 40% to 100% field capacity). We related the accumulated phytotoxic O3 dose (POD1) and cumulative drought index (CDI) to the O3 and drought VFI and assessed growth increment (height, diameter, leaf number), biomass (of all organs), and physiological parameters: net photosynthesis per plant (Pn), photosynthetic nitrogen (PNUE) and phosphorus use efficiency (PPUE)). The results indicated that an increase in POD1 promoted O3 VFI in Quercus robur and Quercus pubescens, while Quercus ilex was asymptomatic. The POD1-based critical level at the onset of O3 VFI was lower for Q. robur than for Q. pubescens (12.2 vs. 15.6 mmol m-2 POD1). Interestingly, drought reduced O3 VFI in Q. robur but increased it in Q. pubescens. Both O3 and drought were detrimental to the plant biomass. However, Q. robur and Q. pubescens invested more in shoots than in roots, while Q. ilex invested more in roots, which might be related to a hormetic mechanism. Pn, PNUE and PPUE decreased in all species under drought, and only in the sensitive Q. robur (PPUE) and Q. pubescens (PNUE) under O3. This study confirms that POD1 is a good indicator to explain the development of O3 VFI and helps a differential diagnosis of co-occurring drought and O3 VFI in oak forests.

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.

Season-long exposure of bilberry plants to realistic and future ozone pollution improves the nutraceutical quality of fruits.

Ozone (O3) is a phytotoxic air pollutant capable of limiting plant yield and growth, and altering the quality of edible plant products. This study aimed to investigate the effects of long-term O3 exposure at realistic and future concentrations (applied during fruit development) not only on morphological, physiological, and biochemical plant/leaf traits of Vaccinium myrtillus but also on its fruit yield and quality. Three-year-old saplings were grown from May to July under three levels of O3 concentration [1.0, 1.5 and 2.0 times the ambient air concentrations, denoted as AA, 1.5_AA and 2.0_AA], using a new-generation O3 Free Air Controlled Exposure system. Ozone induced oxidative pressure and membrane denaturation as confirmed by the accumulation of anion superoxide, hydrogen peroxide (•O2-: +39 and + 29%; H2O2: +55 and + 59% in 1.5_AA and 2.0_AA, respectively, compared with AA), and malondialdehyde by-product (1.4- and 2.5-fold higher than AA, in 1.5_AA and 2.0_AA, respectively). The observed oxidative burst likely affected several cellular structures interested by photosynthetic processes (e.g., decrease of the maximum rate of carboxylation: -30%). This constraint likely induced a decline in plant vitality and a different partitioning of biomass allocation between above and below organs. An accelerated maturation of bilberries due to O3 was reported, suggesting that plants grown under harsher environmental conditions suffered from metabolic changes associated with early ripening. Increasing O3 concentrations might be responsible for an alteration of the ratio between oxidation and reduction processes mechanisms that was followed by a loss of integrity of membranes, so limiting the availability of energy/resources, triggering enzymatic oxidation of phenols to red/purple pigments, and promoting fruit maturation. To the best of our knowledge, this is the first research showing that long-term O3 exposure during bilberry fruit development influenced not only several plant/leaf traits, but also fruit nutraceutical quality at the time of harvest.

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.