Impact of coppicing on microclimate and understorey vegetation diversity in an ancient Mediterranean oak forest.

Coppicing is one of the oldest silvicultural practices and is still widely applied to produce renewable energy from broadleaf forests. However, the consequences on microclimate and understorey vegetation are still poorly understood, especially in Mediterranean oak forests. With the ongoing changes in the climate system and global biodiversity loss, a better understanding of how the forest temperature buffering capacity and below-canopy plant community are impacted by coppicing is crucial. Here we quantify microclimate and understorey vegetation changes in adjacent ancient coppice-with-standards and high forest stands dominated by oaks in Italy, where these systems have been applied for a long time. Air and soil temperatures were recorded for 2.5 years, and nested vegetation plots were used to analyse coppicing effects on species composition, taxonomic, phylogenetic, and functional diversity. Coppicing significantly reduced the forest temperature buffering capacity. The mean of the daily maximum temperatures over the entire period was 1.45 °C higher in the coppiced sites, whereas the mean of the daily minimum temperatures was 0.62 °C lower than in the high forest. Coppicing increased understorey species richness by favouring generalist taxa, but significantly decreased the proportion of forest specialists. The understorey community in coppiced forests consisted of more warm-adapted species. Moreover, coppicing also led to a loss of phylogenetic evenness and to shifts in diversity and community weighted mean Leaf Dry Matter content, pointing to habitat filtering and acclimation processes. In sum, we show that coppicing affects microclimate and understory vegetation in a direction that can exacerbate the effects of climate change, negatively affecting the oak forest specialist flora and its phylogenetic evenness.

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.

Edge effects on the realised soil seed bank along microclimatic gradients in temperate European forests.

Despite the crucial role of the seed bank in forest conservation and dynamics, the effects of forest edge microclimate and climate warming on germination responses from the forest seed bank are still almost unknown. Here, we investigated edge effects on the realised seed bank and seedling community in two types of European temperate deciduous forest, one in the Oceanic and one in the Mediterranean climatic region. Responses in terms of seedling density, diversity, species composition and functional type of the seed bank at the forest edge and interior were examined along latitudinal, elevational and stand structural gradients by means of soil translocation experiments. Moreover, we translocated soil samples from high to low elevation forests in the two regions, thus performing a warming simulation. Density, species diversity and mortality of the seedlings varied with region and elevation. Seedling density also differed between forest edge and interior position, while seedling cover mainly depended on forest structure. Both the edge and interior forest seed bank contained a high proportion of generalist species. In Belgium, a more homogeneous seed bank was found at the forest edge and interior, while in Italy compositional and ecological differences were larger: at the forest edge, more light and less moisture demanding seedling communities developed, with a higher proportion of generalists compared to the interior. In both regions, the upland-to-lowland translocation experiment revealed effects of warming on forest seed banks with thermophilization of the realised communities. Moreover, edge conditions shifted the seedling composition towards more light-demanding communities. The establishment of more light and warm-adapted species from the seed bank could in the long term alter the aboveground vegetation composition, with communities becoming progressively richer in light-demanding generalists and poorer in forest specialists.

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.

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.

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.

Ozone impairs the response of isoprene emission to foliar nitrogen and phosphorus in poplar.

Tropospheric ozone (O3) impairs physiological processes of plants while nitrogen (N) deposition may cause imbalances in soil N and other nutrients such as phosphorus (P) suggesting an increase of P demand for plants. However, the combined effect of O3, soil N and P on isoprene emission from leaves has never been tested. We therefore examined isoprene emission in leaves of Oxford poplar clone exposed to O3 (ambient, AA [35.0 nmol mol-1 as daily mean]; 1.5 × AA; 2.0 × AA), soil N (0 and 80 kg N ha-1) and soil P (0, 40 and 80 kg P ha-1) in July and September in a Free-Air Controlled Exposure (FACE) facility. We also investigated the response of isoprene emission to foliar N, P and abscisic acid (ABA) contents in September because the 2-C-methylerythritol-5-phosphate (MEP) pathway of isoprenoid biosynthesis produces ABA. We found that O3 increased isoprene emission in July, which was associated to increased dark respiration, suggesting an activation of metabolism against O3 stress as an initial response. However, O3 decreased isoprene emission in September which was associated to reduced net photosynthesis. In September, isoprene emission was positively correlated with leaf N content and negatively correlated with leaf P content in AA. However, no response of isoprene emission to foliar N and P was found in elevated O3, suggesting that the isoprene responses to foliar N and P depended on the O3 exposure levels. Isoprene emission rate in 1.5 × AA and 2.0 × AA increased with increasing leaf ABA content, indicating accelerated senescence of injured leaves to favor new leaf growth when high O3 and nutritional availability in the soil were combined. Even though foliar N and P usually act as a proxy for isoprene emission rate, the impact of recent abiotic factors such as O3 should be always considered for modeling isoprene emission under climate change.

Ozone-induced impairment of night-time stomatal closure in O3-sensitive poplar clone is affected by nitrogen but not by phosphorus enrichment.

Nocturnal transpiration may be a key factor influencing water use in plants. Tropospheric ozone (O3) and availability of nutrients such as nitrogen (N) and phosphorus (P) in the soil can affect daytime water use through stomata, but the combined effects of O3, N and P on night-time stomatal conductance (gs) are not known. We investigated the effects of O3 and soil availability of N and P on nocturnal gs and the dynamics of stomatal response after leaf severing in an O3-sensitive poplar clone (Oxford) subjected to combined treatments over a growing season in an O3 free air controlled exposure (FACE) facility. The treatments were two soil N levels (0 and 80 kg N ha-1; N0 and N80), three soil P levels (0, 40 and 80 kg P ha-1; P0, P40 and P80) and three O3 levels (ambient concentration, AA [35.0 ppb as hourly mean]; 1.5 × AA; 2.0 × AA). The analysis of stomatal dynamics after leaf severing suggested that O3 impaired stomatal closure execution. As a result, nocturnal gs was increased by 2.0 × AA O3 in August (+39%) and September (+108%). Night-time gs was correlated with POD0 (phytotoxic O3 dose) and increased exponentially after 40 mmol m-2 POD0. Such increase of nocturnal gs was attributed to the emission of ethylene due to 2.0 × AA O3 exposure, while foliar abscisic acid (ABA) or indole-3-acetic acid (IAA) did not affect gs at night. Interestingly, the O3-induced stomatal opening at night was limited by N treatments in August, but not limited in September. Phosphorus decreased nocturnal gs, although P did not modify the O3-induced stomatal dysfunction. The results suggest that the increased nocturnal gs may be associated with a need to improve N acquisition to cope with O3 stress.

Estimation of the Allergenic Potential of Urban Trees and Urban Parks: Towards the Healthy Design of Urban Green Spaces of the Future.

The impact of allergens emitted by urban green spaces on health is one of the main disservices of ecosystems. The objective of this work is to establish the potential allergenic value of some tree species in urban environments, so that the allergenicity of green spaces can be estimated through application of the Index of Urban Green Zones Allergenicity (IUGZA). Multiple types of green spaces in Mediterranean cities were selected for the estimation of IUGZ. The results show that some of the ornamental species native to the Mediterranean are among the main causative agents of allergy in the population; in particular, Oleaceae, Cupressaceae, Fagaceae, and Platanus hispanica. Variables of the strongest impact on IUGZA were the bioclimatic characteristics of the territory and design aspects, such as the density of trees and the number of species. We concluded that the methodology to assess the allergenicity associated with urban trees and urban areas presented in this work opens new perspectives in the design and planning of urban green spaces, pointing out the need to consider the potential allergenicity of a species when selecting plant material to be used in cities. Only then can urban green areas be inclusive spaces, in terms of public health.

The passion fruit liana (Passiflora edulis Sims, Passifloraceae) is tolerant to ozone.

Passiflora edulis Sims is a liana species of high economic interest and is an interesting model plant for understanding ozone action on disturbed vegetation. In this work we hypothesized that P. edulis has adaptive responses to oxidative stress that enable it to tolerate ozone damage based on its capacity to grow under a diversity of environmental conditions and to dominate disturbed areas. We exposed seedlings to three levels of ozone in a Free-Air Controlled Exposure (FACE) system (22, 41 and 58 ppb h AOT40 and 13.52, 17.24 and 20.62 mmol m-2 POD0, over 97 days) for identifying its tolerance mechanisms. Anatomical (leaf blade structure and fluorescence emission of chloroplast metabolites), physiological (leaf gas exchange, growth rate and biomass production) and biochemical (pigments, total sugars, starch, enzymatic and non-enzymatic antioxidant metabolites, reactive oxygen species and lipid peroxidation derivatives) responses were assessed. Ozone caused decreased total number of leaves, hyperplasia and hypertrophy of the mesophyll cells, and accelerated leaf senescence. However, O3 did not affect carbohydrates content, net photosynthetic rate, or total biomass production, indicating that the carboxylation efficiency and associated physiological processes were not affected. In addition, P. edulis showed higher leaf contents of ascorbic acid, glutathione (as well high ratio between their reduced and total forms), carotenoids, and flavonoids located in the chloroplast outer envelope membrane. Our results indicate that P. edulis is an O3-tolerant species due to morphological acclimation responses and an effective antioxidant defense system represented by non-enzymatic antioxidants, which maintained the cellular redox balance under ozone.

Can nutrient fertilization mitigate the effects of ozone exposure on an ozone-sensitive poplar clone?

We tested the independent and interactive effects of nitrogen (N; 0 and 80 kg ha-1), phosphorus (P; 0, 40 and 80 kg ha-1), and ozone (O3) application/exposure [ambient concentration (AA), 1.5 × AA and 2.0 × AA] for five consecutive months on biochemical traits of the O3-sensitive Oxford poplar clone. Plants exposed to O3 showed visible injury and an alteration of membrane integrity, as confirmed by the malondialdehyde by-product accumulation (+3 and +17% under 1.5 × AA and 2.0 × AA conditions, in comparison to AA). This was probably due to O3-induced oxidative damage, as documented by the production of superoxide anion radical (O2-, +27 and +63%, respectively). Ozone per se, independently from the concentrations, induced multiple signals (e.g., alteration of cellular redox state, increase of abscisic acid/indole-3-acetic acid ratio and reduction of proline content) that might be part of premature leaf senescence processes. By contrast, nutrient fertilization (both N and P) reduced reactive oxygen species accumulation (as confirmed by the decreased O2- and hydrogen peroxide content), resulting in enhanced membrane stability. This was probably due to the simultaneous involvement of antioxidant compounds (e.g., carotenoids, ascorbate and glutathione) and osmoprotectants (e.g., proline) that regulate the detoxification processes of coping with oxidative stress by reducing the O3 sensitivity of Oxford clone. These mitigation effects were effective only under AA and 1.5 × AA conditions. Nitrogen and P supply activated a free radical scavenging system that was not able to delay leaf senescence and mitigate the adverse effects of a general peroxidation due to the highest O3 concentrations.

Cross-talk between physiological and biochemical adjustments by Punica granatum cv. Dente di cavallo mitigates the effects of salinity and ozone stress.

Plants are exposed to a broad range of environmental stresses, such as salinity and ozone (O3), and survive due to their ability to adjust their metabolism. The aim of this study was to evaluate the physiological and biochemical adjustments adopted by pomegranate (Punica granatum L. cv. Dente di cavallo) under realistic field conditions. One-year-old saplings were exposed to O3 [two levels denoted as ambient (AO) and elevated (EO) O3 concentrations] and salinity [S (salt, 50 mM NaCl)] for three consecutive months. No salt (NS) plants received distilled water. Despite the accumulation of Na+ and Cl- in the aboveground biomass, no evidence of visible injury due to salt (e.g. tip yellow-brown lesions) was found. The maintenance of leaf water status (i.e. unchanged values of electrolytic leakage and relative water content), the significant increase of abscisic acid, proline and starch content (+98, +65 and +59% compared to AO_NS) and stomatal closure (-24%) are suggested to act as adaptive mechanisms against salt stress in AO_S plants. By contrast, EO_NS plants were unable to protect cells against the negative impact of O3, as confirmed by the reduction of the CO2 assimilation rate (-21%), accumulation of reactive oxygen species (+10 and +225% of superoxide anion and hydrogen peroxide) and malondialdehyde by-product (about 2-fold higher than AO_NS). Plants tried to preserve themselves from further oxidative damage by adopting some biochemical adjustments [i.e. increase in proline content (+41%) and induction of catalase activity (8-fold higher than in AO_NS)]. The interaction of the two stressors induced responses considerably different to those observed when each stressor was applied independently. An analysis of the antioxidant pool revealed that the biochemical adjustments adopted by P. granatum under EO_S conditions (e.g. reduction of total ascorbate; increased activities of superoxide dismutase and catalase) were not sufficient to ameliorate the O3-induced oxidative stress.

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.

Should we see urban trees as effective solutions to reduce increasing ozone levels in cities?

Outdoor air pollution is considered as the most serious environmental problem for human health, associated with some million deaths worldwide per year. Cities have to cope with the challenges due to poor air quality impacting human health and citizen well-being. According to an analysis in the framework of this study, the annual mean concentrations of tropospheric ozone (O3) have been increasing by on average 0.16 ppb year-1 in cities across the globe over the time period 1995-2014. Green urban infrastructure can improve air quality by removing O3. To efficiently reduce O3 in cities, it is important to define suitable urban forest management, including proper species selection, with focus on the removal ability of O3 and other air pollutants, biogenic emission rates, allergenic effects and maintenance requirements. This study reanalyzes the literature to i) quantify O3 removal by urban vegetation categorized into trees/shrubs and green roofs; ii) rank 95 urban plant species based on the ability to maximize air quality and minimize disservices, and iii) provide novel insights on the management of urban green spaces to maximize urban air quality. Trees showed higher O3 removal capacity (3.4 g m-2 year-1 on average) than green roofs (2.9 g m-2 year-1 as average removal rate), with lower installation and maintenance costs (around 10 times). To overcome present gaps and uncertainties, a novel Species-specific Air Quality Index (S-AQI) of suitability to air quality improvement is proposed for tree/shrub species. We recommend city planners to select species with an S-AQI>8, i.e. with high O3 removal capacity, O3-tolerant, resistant to pests and diseases, tolerant to drought and non-allergenic (e.g. Acer sp., Carpinus sp., Larix decidua, Prunus sp.). Green roofs can be used to supplement urban trees in improving air quality in cities. Urban vegetation, as a cost-effective and nature-based approach, aids in meeting clean air standards and should be taken into account by policy-makers.

Ozone risk assessment is affected by nutrient availability: Evidence from a simulation experiment under free air controlled exposure (FACE).

Assessing ozone (O3) risk to vegetation is crucial for informing policy making. Soil nitrogen (N) and phosphorus (P) availability could change stomatal conductance which is the main driver of O3 uptake into a leaf. In addition, the availability of N and P could influence photosynthesis and growth. We thus postulated that the sensitivity of plants to O3 may be changed by the levels of N and P in the soil. In this study, a sensitive poplar clone (Oxford) was subject to two N levels (N0, 0 kg N ha-1; N80, 80 kg N ha-1), three P levels (P0, 0 kg P ha-1; P40, 40 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) for a whole growing season in an O3 free air controlled exposure (FACE) facility. Flux-based (POD0 to 6) and exposure-based (W126 and AOT40) dose-response relationships were fitted and critical levels (CLs) were estimated for a 5% decrease of total annual biomass. It was found that N and P availability modified the dose-response relationships of biomass responses to O3. Overall, the N supply decreased the O3 CLs i.e. increased the sensitivity of poplar to O3. Phosphorus alleviated the O3-caused biomass loss and increased the CL. However, such mitigation effects of P were found only in low N and not in high N conditions. In each nutritional treatment, similar performance was found between flux-based and exposure-based indices. However, the flux-based approach was superior, as compared to exposure indices, to explain the biomass reduction when all nutritional treatments were pooled together. The best O3 metric for risk assessments was POD4, with 4.6 mmol m-2 POD4 as a suitable CL for Oxford poplars grown under various soil N and P conditions.

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.

Testing a ratio of photosynthesis to O3 uptake as an index for assessing O3-induced foliar visible injury in poplar trees.

Visible foliar injury by ozone (ozone visible injury) is known as a biomarker to assess potential phytotoxicity of ozone. We investigated ozone visible injury in an ozone-sensitive poplar (Oxford clone) under a 2-year free-air controlled exposure (FACE) experiment and calculated three ozone indices (i.e., accumulative ozone exposure over 40 ppb during daylight hours (AOT40), phytotoxic ozone dose above a flux threshold of 0 nmol m-2 s-1 (POD0), and the cumulative value of the ratio of hourly ozone uptake to net photosynthesis (ΣU/P n ) to assess the critical level (CL) at the time of the first symptom onset of ozone visible injury. We tested the hypothesis that ozone injury depends both on the amount of ozone entering a leaf and on the capacity for biochemical detoxification or repair with photosynthesis as a proxy. The CLs at the time of the first symptom onset of ozone visible injury were 19 ppm h for AOT40, 26 mmol m-2 for POD0, and 1.2 mol mol-1 for ΣU/P n in Oxford clone at the ozone FACE experiment. Our findings were then verified by 4-year observation-based data in central Italy on Oxford clone and white poplar (Populus alba L.). These observation-based data indicated that we found ozone visible injury in Oxford clone even though AOT40 was relatively low (11.7 ppm h). On the other hand, when values of POD0 and ΣU/P n exceeded over the CLs, the occurrence of initial symptoms in Oxford clone was shown. White poplar did not show ozone visible injury. ΣU/P n of white poplar at the field sites reached ~1.0 mol mol-1 (less than the CL = 1.2 mol mol-1, which was obtained from O3 FACE) during May-September, although the values of POD0 were relatively high in white poplar (44-47 mmol m-2 during May-September). The result implies that ozone injury may have occurred in poplars when stomatal ozone flux exceeded the critical range of tolerance due to the assimilate shortage for repair and defense against ozone stress.

Responses of serpentine plants to pine invasion: Vegetation diversity and nickel accumulation in species with contrasting adaptive strategies.

Introduction of non-native trees is one of the major threats to ecosystem integrity and biodiversity. Stands of maritime pine (Pinus pinaster Ait.) introduced decades ago represent a threat to the specialized plant communities of serpentine outcrops in Italy. This study investigates the effects of such invasions at the community and species level, based on vegetation sampling in three selected sites with comparable environmental conditions. Pine cover caused a decrease of α-diversity by lowering the species evenness of the community, though species richness was not negatively affected. Compositional changes between the two habitats were significant but not clearly associated with a decrease in taxonomic distinctness in the pine stands. As many as nine indicator species were found in the open vegetation, along with the obligate endemics Odontarrhena bertolonii and Armeria denticulata. Both of them declined in the pine stands. Here, an increase in the phytoavailable nickel fraction was associated with a decrease in total nickel concentration in the soil, via mobilization of the metal caused by lowering of pH induced by the conifer litter. The nickel-hyperaccumulator O. bertolonii was able to maintain high metal concentrations in the shoots despite a decrease in root concentration, resulting in a higher shoot/root ratio in the pine stands (~20). Conversely, shoot/root ratio in the non-accumulator Plantago holosteum was <1 and not affected by the conifer, as well as its abundance in this anthropogenic habitat. Contrasting responses of the two species were likely due to their different sensitivity to modified light and soil conditions, whereas stability of shoot nickel-concentration in O. bertolonii did not support increased predation by natural enemies as one of the causes for its decline under the conifer. Progressive thinning of these stands is advocated to limit soil nickel mobilization and to restore a unique ecosystem with its endemic metallophytes.