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.

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.

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.

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.

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.

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.

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.

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.

Metabolic and physiological alterations indicate that the tropical broadleaf tree Eugenia uniflora L. is sensitive to ozone.

Eugenia uniflora L. is an important fruit tree native to tropical South America that adapts to different habitats, thanks to its metabolic diversity and ability to adjust the leaf antioxidant metabolism. We hypothesized that this metabolic diversity would also enable E. uniflora to avoid oxidative damage and tolerate the enhanced ozone (O3) concentrations that have been registered in the (sub)tropics. We investigated whether carbohydrates, polyphenols and antioxidants are altered and markers of oxidative damage (ROS accumulation, alterations in leaf gas exchange, growth and biomass production) are detected in plants exposed to two levels of O3 (ambient air and twice elevated ozone level in a O3-FACE system for 75 days). Phytotoxic O3 dose above a threshold of 0 nmol m-2 s-1 (POD0) and accumulated exposure above 40 ppb (AOT40) were 3.6 mmol m-2 and 14.898 ppb h at ambient, and 4.7 mmol m-2 and 43.881 ppb h at elevated O3. Twenty-seven primary metabolites and 16 phenolic compounds were detected in the leaves. Contrary to the proposed hypothesis that tropical broadleaf trees are relatively O3 tolerant, we concluded that E. uniflora plants are sensitive to elevated O3 concentrations. Experimental POD0 values were lower than the critical levels for visible foliar O3, because of low stomatal conductance. In spite of this low stomatal O3 uptake, we found classic O3 injury, e.g. reduction in carbohydrates and fatty acids concentrations; non-significant changes in the polyphenol profile; inefficient antioxidant responses; increased contents of ROS and indicators of lipid peroxidation; reductions in stomatal conductance, net photosynthesis, root/shoot ratio and height growth. However, we also found some compensation mechanisms, e.g. increased leaf concentration of polyols for protecting the membranes, and increased leaf number for compensating the decline of photosynthetic rate. These results help filling the knowledge gap about tropical tree responses to O3.

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.

Economic impacts of ambient ozone pollution on wood production in Italy.

Worldwide, tropospheric ozone (O3) is a potential threat to wood production, but our understanding of O3 economic impacts on forests is still limited. To overcome this issue, we developed an approach for integrating O3 risk modelling and economic estimates, by using the Italian forests as a case study. Results suggested a significant impact of O3 expressed in terms of stomatal flux with an hourly threshold of uptake (Y = 1 nmol O3 m-2 leaf area s-1 to represent the detoxification capacity of trees), i.e. POD1. In 2005, the annual POD1 averaged over Italy was 20.4 mmol m-2 and the consequent potential damage ranged from 790.90 M€ to 2.85 B€ of capital value (i.e. 255-869 € ha-1, on average) depending on the interest rate. The annual damage ranged from 31.6 to 57.1 M€ (i.e. 10-17 € ha-1 per year, on average). There was also a 1.1% reduction in the profitable forest areas, i.e. with a positive Forest Expectation Value (FEV), with significant declines of the annual national wood production of firewood (- 7.5%), timber pole (- 7.4%), roundwood (- 5.0%) and paper mill (- 4.8%). Results were significantly different in the different Italian regions. We recommend our combined approach for further studies under different economic and phytoclimatic conditions.