Effects of elevated O3, alone and in combination with elevated CO2, on tree leaf chemistry and insect herbivore performance: a meta-analysis

We reviewed the effects of elevated ozone (O₃), alone and in combination with elevated carbon dioxide (CO₂) on primary and secondary metabolites of trees and performance of insect herbivores by means of meta‐analysis. Our database consisted of 63 studies conducted on 22 species of trees and published between 1990 and 2005. Ozone alone had no overall effect on concentrations of carbohydrates or nutrients, whereas in combination with CO₂, elevated O₃ reduced nutrient concentrations and increased carbohydrate concentrations. In contrast to primary metabolites, concentrations of phenolics and terpenes were significantly increased by 16% and 8%, respectively, in response to elevated O₃. Effects of ozone in combination with elevated CO₂ were weaker than those of ozone alone on phenolics, but stronger than those of ozone alone on terpenes. The magnitude of secondary metabolite responses depended on the type of ozone exposure facility and increased in the following order: indoor growth chamber 3 than gymnosperms, as shifts in concentrations of carbohydrate and phenolics were observed in the former, but not in the latter. Elevated O₃ had positive effects on some indices of insect performance: pupal mass increased and larval development time shortened, but these effects were counteracted by elevated CO₂. Therefore, despite the observed increase in secondary metabolites, elevated O₃ tends to increase tree foliage quality for herbivores, but elevated CO₂ may alleviate these effects. Our meta‐analysis clearly demonstrated that effects of elevated O₃ alone on leaf chemistry and some indices of insect performance differed from those of O₃+CO₂, and therefore, it is important to study effects of several factors of global climate change simultaneously.     

Psidium guajava Paluma responses to environmental conditions and ozone concentrations in the urban forest of São Paulo,SE-Brazil

Tropospheric ozone concentrations (O₃) are continuously increasing with harmful effects on vegetation. Ozone uptake triggers oxidative stress that may promote adverse changes in various plant physiological processes. The aim of this study was to evaluate the relationship between meteorological conditions and O₃ exposure in foliar injuries, gas exchanges and antioxidant enzyme activity in Psidium guajava Paluma. Saplings of Paluma were exposed in a site contaminated by O_3. Foliar injuries, gas exchanges, and the activity of superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR) were evaluated weekly. Results were analyzed by multivariate analysis. The O₃ exposure reduced carbon assimilation and induced the onset of foliar injuries. The enzymatic activities were more related to meteorological conditions than to O₃ exposure, indicating that these enzymes are not this species’ first line of defense. However, our finding indicates that Paluma may be considered a response indicator, given its presentation of foliar injuries even at low O₃ concentrations.     

Ozone mediators effect on “in vitro” scratch wound closure

The beneficial effect of low doses of ozone on wound healing has been well documented and attributed mainly to its bactericidal and pro-oxidant properties. Because ozone itself does not penetrate the cells but immediately reacts with polyunsaturated fatty acids, its effects are the results of oxidative mediators. Among the molecule produces by the interaction of ozone with biological systems, there are HNE and H₂O₂. At today, the cellular mechanisms accounting for the positive effects of mild ozonization on wound closure are still largely unexplored. The aim of the present study was to evaluate the effect of different non-toxic doses of ozonated saline ranging from 2 to 300?μM, in an in vitro wound scratch model by the use of human keratinocytes. The results showed that ozonated saline is able to improve in vitro wound healing by stimulating cell proliferation as measured by BrdU assay and PCNA protein levels. In order to better elucidate the molecules that play the main role in the beneficial effect of ozonated saline in wound healing, HNE and H₂O₂ were used alone or in combination to mimic ozonated saline effect. Surprisingly, keratinocytes treated with different doses of HNE and H₂O₂ did not significantly improve the wound closure, while the combination of the two compounds was able to improve wound closure. In addition, Nrf2 pathways were also activated as determined by its translocation to the nucleus and the increased HO1 gene expression. The present work suggests that ozonated saline effect on wound closure is the results of the combination of more molecules among which HNE and H₂O₂ play a key role.     

Characterization of Gaseous Ozone Decomposition in Soil

Laboratory scale batch experiments were performed to investigate the decomposition characteristics of gaseous ozone in porous media. The decomposition rates of gaseous ozone in several solid media were determined, and the relationship of moisture content with sorbed ozone molecules was evaluated. Ozone decomposition in control and glass beads packed columns followed second-order reaction kinetics, while ozone consumption in a sand-packed column demonstrated first-order kinetics with a rate constant of 0.0109 min^?1 and half-life of 1.0 h. The presence of typical metal oxides in the soil resulted in ozone consumption rates in the following order: hematite (Fe₂O₃) > silica-alumina (SiO₂Al₂O₃) > alumina (Al₂O₃) > silica (SiO₂). Ozone decomposition was highly dependent upon soil moisture content. Over 90% of the total ozone mass decomposed in the field soil with moisture content at less than 1 wt%, whereas as low as 5–15% of the total ozone mass degraded with moisture content at more than 2 wt%. In conclusion, ozone decomposition in soils was primarily controlled not only by soil organic matter but also by reactive metal oxides on the soil surface. These two factors were shown to be highly dependent upon soil moisture content.     

Flixweed Is More Competitive than Winter Wheat under Ozone Pollution: Evidences from Membrane Lipid Peroxidation,Antioxidant Enzymes and Biomass

To investigate the effects of ozone on winter wheat and flixweed under competition, two species were exposed to ambient, elevated and high [O₃] for 30 days, planted singly or in mixculture. Eco-physiological responses were examined at different [O₃] and fumigating time. Ozone reduced the contents of chlorophyll, increased the accumulation of H₂O₂ and malondialdehyde in both wheat and flixweed. The effects of competition on chlorophyll content of wheat emerged at elevated and high [O₃], while that of flixweed emerged only at high [O₃]. The increase of H₂O₂ and malondialdehyde of flixweed was less than that of wheat under the same condition. Antioxidant enzyme activities of wheat and flixweed were seriously depressed by perennial and serious treatment using O₃. However, short-term and moderate fumigation increased the activities of SOD and POD of wheat, and CAT of flixweed. The expression levels of antioxidant enzymes related genes provided explanation for these results. Furthermore, the increase of CAT expression of flixweed was much higher than that of SOD and POD expression of wheat. Ozone and competition resulted in significant reductions in biomass and grain yield in both winter wheat and flixweed. However, the negative effects on flixweed were less than wheat. Our results demonstrated that winter wheat is more sensitive to O₃ and competition than flixweed, providing valuable data for further investigation on responses of winter wheat to ozone pollution, in particular combined with species competition.     

Response of photosynthesis and chlorophyll fluorescence to acute ozone stress in tomato (Solanum lycopersicum Mill.)

The crop sensitivity to ozone (O₃) is affected by the timing of the O₃ exposure, by the O₃ concentration, and by the crop age. To determine the physiological response to the acute ozone stress, tomato plants were exposed to O₃ at two growth stages. In Experiment I (Exp. I), O₃ (500 μg m^?3) was applied to 30-d-old plants (PL30). In Experiment II (Exp. II), three O₃ concentrations (200, 350, and 500 μg m^?3) were applied to 51-d-old plants (PL51). The time of the treatment was 4 h (7:30–11:30 h). Photosynthesis and chlorophyll fluorescence measurements were done 4 times (before the exposure; 20 min, 20 h, and 2–3 weeks after the end of the treatment) using a LI-COR 6400 photosynthesis meter. The stomatal pore area and stomatal conductance were reduced as the O₃ concentration increased. Ozone induced the decrease in the photosynthetic parameters of tomato regardless of the plant age. Both the photosystem (PS) II operating efficiency and the maximum quantum efficiency of PSII photochemistry declined under the ozone stress suggesting that the PSII activity was inhibited by O₃. The impaired PSII contributed to the reduced photosynthetic rate. The greater decline of photosynthetic parameters was found in the PL30 compared with the PL51. It proved the age-dependent ozone sensitivity of tomato, where the younger plants were more vulnerable. Ozone caused the degradation of photosynthetic apparatus, which affected the photosynthesis of tomato plants depending on the growth stage and the O₃ concentration.     

臭氧胁迫对水稻生长以及C、N、S元素分配的影响

采用开顶式气室(Open-top Chamber, OTC),对水稻"3694繁"(Oryza sativa L., 3694 Fan)在浙江嘉兴进行田间原位臭氧(O₃)熏气实验,研究不同臭氧浓度熏气对水稻生长以及C、N,S元素分配的影响。实验设置分4个水平:过滤大气组(CF,10 nL/L)、自然大气组(NF,40 nL/L)和两个不同浓度的臭氧处理组(O₃-1:100 nL/L; O₃-2:150 nL/L)。主要结果表明:(1)开始臭氧熏气时,各个处理组单茎水稻各组分生物量没有差异. 在熏气后期(水稻成熟期),臭氧处理使单茎水稻根、茎和穗生物量显著下降,根冠比降低,株高显著降低,表明臭氧胁迫增加水稻地上部分的干物质分配,且对株高的影响可能大于对地上生物量的影响;(2)臭氧处理使水稻根和茎C元素含量下降,叶C元素含量上升,表明臭氧胁迫提高了叶片中碳分配,而降低了根和茎的碳分配;(3)各个组分N元素含量上升和碳氮比下降;(4)茎、叶和穗S元素含量上升,可能会增强水稻抗氧化系统的作用,从而抵抗臭氧胁迫。所有实验结果表明臭氧浓度升高会对水稻生长产生严重不利影响,从而导致水稻各个组分的C、N、S元素分配格局发生改变。     

How does elevated ozone impact soybean? A meta‐analysis of photosynthesis,growth and yield

Surface ozone concentrations ([O₃]) during the growing season in much of the northern temperate zone reach mean peak daily concentrations of 60 p.p.b. Concentrations are predicted to continue to rise over much of the globe during the next 50 years. Although these low levels of ozone may not induce visible symptoms on most vegetation, they can result in substantial losses of production and reproductive output. Establishing the vulnerability of vegetation to rising background ozone is complicated by marked differences in findings between individual studies. Ozone effects are influenced by exposure dynamics, nutrient and moisture conditions, and the species and cultivars that are investigated. Meta‐analytic techniques provide an objective means to quantitatively summarize treatment responses. Soybean has been the subject of many studies of ozone effects. It is both the most widely planted dicotyledonous crop and a model for other C₃ annual plants. Meta‐analytic techniques were used to quantitatively summarize the response of soybean to an average, chronic ozone exposure of 70 p.p.b., from 53 peer‐reviewed studies. At maturity, the average shoot biomass was decreased 34% and seed yield was 24% lower. Even in studies where [O₃] was < 60 p.p.b., there was a significant decrease in biomass and seed production. At low [O₃], decreased production corresponded to a decrease in leaf photosynthesis, but in higher [O₃] the larger loss in production was associated with decreases in both leaf photosynthesis and leaf area. The impact of ozone increased with developmental stage, with little effect on vegetative growth and the greatest effect evident at completion of seed filling. Other stress treatments, including UV‐B and drought, did not alter the ozone response. Elevated carbon dioxide significantly decreased ozone‐induced losses, which may be explained by a significant decrease in stomatal conductance.     

不同秧苗素质和移栽密度条件下臭氧胁迫对水稻光合作用、物质生产和产量的影响

近地层臭氧浓度升高使水稻生长受抑进而使产量下降,但这种影响是否因不同栽培条件而异尚不清楚。2011年依托先进的稻田臭氧FACE(Free Air gas Concentration Enrichment)技术平台,以汕优63为供试材料,臭氧设置大气臭氧浓度(Ambient)和高臭氧浓度(比Ambient高50%),秧苗素质设置弱苗(移栽时无分蘖)和壮苗(移栽时带两个分蘖),移栽密度设置低密度(16穴/m2)、中密度(24穴/m2)和高密度(32穴/m2),研究不同秧苗素质和移栽密度条件下臭氧胁迫对水稻生长和产量的影响。结果表明:高浓度臭氧使水稻结实期叶片SPAD值、净光合速率、气孔导度和蒸腾速率明显下降,但胞间CO2浓度和叶温无显著变化。高浓度臭氧对水稻拔节前物质生产量没有影响,但使拔节至抽穗期、抽穗至成熟期物质生产量平均分别降低13%和29%,进而使成熟期生物产量和籽粒产量均显著下降。方差分析表明,臭氧与秧苗素质间没有互作效应,但臭氧与移栽密度的互作对最终产量的影响达显著水平。以上结果表明,臭氧胁迫使水稻生长后期光合受阻,导致物质生产和产量显著下降;适当增加移栽密度可能会减少臭氧胁迫下水稻产量的损失。     

CO2 photoassimilation and chlorophyll fluorescence in two clover species showing different response to O3

The current study evaluated photosynthetic processes in two clover species based on their performance under high O₃ conditions (150 nl l^–1 for 3 h). These species are Trifolium repens L. and Trifolium pratense L., which are well known for their different sensitivity to ozone. Ozone affected the two clover species very differently. In T. pratense, ozone induced visible symptoms of damage even though CO₂ photoassimilation, stomatal conductance and the electron transport rate were not affected. A decrease in the optimal quantum yield was observed in T. pratense immediately after the end of the period of O₃ stress but it reverted to a value similar to the control 24 h after removing the stress, indicating that non-irreversible photoinhibition had occurred. Data obtained for symptomatic T. pratense indicate that still-green living tissue of the leaf is able to carry out CO₂ assimilation; the only parameter found to be affected by O₃ was the efficiency of excitation capture. In T. repens, acute O₃ fumigation induced inhibition of photosynthetic activity, enhanced stomatal closure and increased the reduction state of the PSII primary acceptor. A possible explanation for the inhibition of photosynthesis could reside in inhibition of the Calvin cycle over-reducing PSII, thus increasing (1 – q_P) and increasing non-photochemical quenching.     

Interaction of ozone pollution and light effects on photosynthesis in a forest canopy experiment

Ozone pollution may reduce net carbon gain in forests, yet data from mature trees are rare and the effects of irradiance on the response of photosynthesis to ozone remain untested. We used an open-air system to expose 10 branches within the upper canopy of an 18-m-tall stand of sugar maple (Acer saccharum Marsh.) to twice-ambient concentrations of ozone (95nmol mol^?1, 0900 to 1700, 1 h mean) relative to 10 paired, untreated controls (45nmol mol^?1) over 3 months. The branch pairs were selected along a gradient from relatively high irradiance (PPFD 14.5 mol m^?2 d^?1) to deep shade (0.7mol m^?2 d^?1). Ozone reduced light-saturated rates of net photosynthesis (A_sat) and increased dark respiration by as much as 56 and 40%, respectively. Compared to sun leaves, shade leaves exhibited greater proportional reductions in A_sat and had lower chlorophyll concentrations, quantum efficiencies, and leaf absorptances when treated with ozone relative to controls. With increasing ozone dose over time, A_sat became uncoupled from stomatal conductance as ratios of internal to external concentrations of carbon dioxide increased, reducing water-use efficiency. Ozone reduced net photosynthesis and impaired stomatal function, with these effects depending on the irradiance environment of the canopy leaves. Increased ozone sensitivity of shade leaves compared to sun leaves has consequences for net carbon gain in canopies.     

Inhibition of the photosynthetic capacity of isolated chloroplasts by ozone

Isolated spinach chloroplasts have been used as a model system for studying the interaction of ozone, a component of photochemical smog, with plant membranes. Ozone bubbled into a suspension of isolated chloroplasts inhibits electron transport in both photosystems without uncoupling ATP production. Photosystem I (reduced 2,6-dichlorophenolindolphenol → NADP⁺) is a little more sensitive than photosystem II (H₂O → 2,6-dichlophenolindolphenol). Ozone does not act as an energy transfer inhibitor, since the drop in ATP production and high energy intermediate (measured by amine-induced swelling) is nearly parallel to the decline in electron transport. A reasonable hypothesis is that ozone disrupts the normal pathway of energy flow from light-excited chlorophyll into the photoacts by a disruption of the components of the membrane but not a general disintegration of the membrane. In addition, ozone does not seem to penetrate into the grana region through the outer membrane of intact plastids, since ozone lowers the bicarbonate-supported O₂ evolution but does not affect the rate of ferricyanide reduction in the same plastids after osmotic disruption. This would indicate that the effect of ozone on green plants, at low concentrations, may be due to the interaction of ozone with the first membrane it contacts and not directly with internal metabolic processes.     

Spatial and temporal variability of ground-level ozone in Castilla-León (Spain)

Ozone is a secondary air pollutant that has received extensive attention in the literature, mainly because of the adverse effects that exposure to it can cause, particularly in vegetation during the growing season. Because meteorological conditions strongly influence the efficiency of photochemical processes leading to ozone formation and destruction, ground-level ozone air pollution is currently being considered as a regional-scale phenomenon rather than a local one. This role of O₃ as a regional-scale pollutant often implies the handling of large data sets in order to obtain information about its spatial and temporal variability patterns over a given broad region. Rotated principal component analysis (RPCA) is known to be one of the most powerful mathematical tools that can be used to achieve this aim. RPCA was applied in this paper to the summer and winter hourly time series of ground-level O₃, concentrations recorded during 2 consecutive years (1996–1997) at 26 urban and suburban sites in Castilla-León (Spain). This procedure permitted the identification of different subregions where O₃ concentrations show different spatio-temporal variability patterns. These variability patterns are mainly associated with the interaction of regional-level meteorological and anthropogenic factors. Some differences between winter and summer patterns were also found. Received: 5 April 1999 / Revised: 27 December 1999 / Accepted: 27 December 1999     

Activation of an oxidative burst is a general feature of sensitive plants exposed to the air pollutant ozone

Ozone exposure stimulates an oxidative burst in leaves of sensitive plants, resulting in the generation and accumulation of hydrogen peroxide (H₂O₂) in tobacco and tomato, and superoxide (O₂^–?) together with H₂O₂ in Arabidopsis accessions. Accumulation of these reactive oxygen species (ROS) preceded the induction of cell death, and both responses co‐occurred spatially in the periveinal regions of the leaves. Re‐current ozone exposure of the sensitive tobacco cv. Bel W3 in closed chambers or in the field led to an enlargement of existing lesions by priming the border cells for H₂O₂ accumulation. Open top chamber experiments with native herbaceous plants in the field showed that Malva sylvestris L. accumulates O₂^–? at those sites that later exhibit plant cell death. Blocking of ROS accumulation markedly reduced ozone‐induced cell death in tomato, Arabidopsis and M. sylvestris. It is concluded that ozone triggers an in planta generation and accumulation of H₂O₂ and/or O₂^–? depending on the species, accession and cultivar, and that both these reactive oxygen species are involved in the induction of cell death in sensitive crop and native plants.     

Ozone-mediated cytotoxicity after short-term exposure and its relation to the production of cellular metabolites (NO,H2O2)

Alveolar macrophages secrete numerous mediators, playing an important role in host defence. Among these mediators, nitric oxide (NO) and hydrogen peroxide (H₂O₂) are both involved in bactericidal killing and trigger the release of other cellular metabolites. We have analyzed the effect of an atmosphere polluted with ozone (0.03–0.5 ppm v/v) on the monocytic cell line THP-1, as a model for alveolar macrophages,in vitro. NO and H₂O₂ were chosen to evaluate cell response to ozone. Cell injury was evaluated using lactate dehydrogenase (LDH) liberation into the medium. An exposure to 0.5 ppm ozone proved to be more toxic to the cells, than 0.1 or 0.03 ppm, evidenced by more LDH being liberated and cytotoxicity reaching values up to 64%. For all ozone concentrations, H₂O₂ production reached a peak value after 10–15 min of exposure, after which the concentration of extracellular H₂O₂ production diminished rapidly. The highest NO concentrations were measured with 0.5 ppm ozone, reaching a maximum value of 1460 nmol/L per 5×10⁶ cells, which is 1.55 times higher than for nonexposed cells. Lower concentrations barely induced higher NO concentrations compared to nonexposed cells. The results indicate that ozone effects not only the viability of human monocytes but also the release of antibacterial and defense signaling molecules and suggest that ozone-mediated cytotoxicity may be related to the secretion of NO and H₂O₂. This revised version was published online in July 2006 with corrections to the Cover Date.     

Similar Effects of Ozone on Four Cultivars of Lettuce in Open Top Chambers During Winter

Ozone is the major phytotoxic air pollutant that reduces the yield of several agricultural crops in the Spanish Mediterranean area. We studied four lettuce cultivars (Lactuca sativa L.) for the effects of different O₃ concentrations during the winter on chlorophyll (Chl) a fluorescence, lipid peroxidation, and root length in outdoor open-top chambers. Under O₃ the photosynthetic quantum conversion declined while heat emissions increased in all cultivars; these results provide more evidence of non-filtered air with additional ozone (NFA+O₃) treatment compared with non-filtered air (NFA) and charcoal filtered ozone-free air (CFA). Changes in the Chl a fluorescence may be associated with an increase in membrane lipid peroxidation as well as with observed reduction of root length under O₃ stress.     

Effect of ozone on spore germination,spore production and biomass production in two <Emphasis Type="Italic">Aspergillus</Emphasis> species

The ability of ozone gas to reduce food spoilage is relatively well documented, but the developmental effects of the gas on food spoilage fungi are not well known. In this study two model aspergilli, Aspergillus nidulans and Aspergillus ochraceus were used to study the effects of ozone on spore germination, sporulation and biomass production. These effects were examined under three levels of ozone; two high level ozone exposures (200 and 300 μmol mol⁻¹) and one low level exposure (0.2 μmol mol⁻¹). The two species behaved noticeably different to each other. Ozone was more effective in reducing growth from spore inocula than mycelia. No spore production could be detected in A. nidulans exposed to continuous low level O₃, whereas the same treatment reduced spores produced in A. ochraceus by 94%. Overall the work suggests that ozone exposure is an effective method to prevent spread of fungal spores in a food storage situation.     

Ozone Inhibition of Photosynthesis in Chlorella sorokiniana

Exposure of Chlorella sorokiniana (07-11-05) to ozone inhibits photosynthesis. In this study, the effects of ozone on O₂ evolution and fluorescence yields are used to characterize this inhibition. At an ozone dose of about 3 micromoles delivered to 2 × 10⁹ cells, the photosynthetic rate of the cells is inhibited 50%, as indicated by a decrease in bicarbonate-stimulated O₂ evolution (control rate, 1.4 ± 0.3 × 10⁻¹⁵ moles per cell per minute).     

Protective effect of ozone treatment on the injury associated with hepatic ischemia-reperfusion: Antioxidant-prooxidant balance

The effects of ozone treatment on the injury associated to hepatic ischemia-reperfusion (I/R) was evaluated. Ozone treatment (1 mg/kg daily during 10 days by rectal insufflation) is shown to be protective as it attenuated the increases in transaminases (AST, ALT) and lactate levels observed after I/R. I/R leads to a decrease in endogenous antioxidant (SOD and glutathione) and an increase in reactive oxygen species (H₂O₂) with respect to the control group. However, ozone treatment results in a preservation (glutathione) or increase (SOD) in antioxidant defense and maintains H₂O₂ at levels comparable to those in the control group. The present study reports a protective effect of ozone treatment on the injury associated to hepatic I/R. The effectiveness of ozone could be related to its action on endogenous antioxidants and prooxidants balance in favour of antioxidants, thus attenuating oxidative stress.     

Influence of acid rain and ozone on soil heavy metals under loblolly pine trees: A field study

A field growth chamber study was conducted to determine the effects of ozone and simulated acid rain (SAR) on soil heavy metals. Loblolly pine (Pinus taeda L.), grown in open-top chambers, was exposed to three concentrations of ozone (charcoal filtered air with 0.026 µL O₃ L^-1, and two non-filtered treatments in which ozone concentrations were 0.074 µL L^-1 and 0.147 µL L^-1, respectively) and two levels of SAR (pH 3.5 and 5.2). Ozone was applied for 12 h d^-1 for 9 months and acid rain deposition was 125 mm event^-1. After 9 months exposure, soil pH, organic matter and DTPA-extractable heavy metals (Cd, Pb, Zn, Mn, Fe, Cu) were determined on soil samples collected from exposed chambers at two depths (0–15 cm and 15–30 cm). Simulated acid rain decreased the original soil pH. The concentrations of Cd, Pb and Mn at SAR pH 3.5 were significantly higher than at SAR pH 5.2. Ozone did not affect Zn, Fe and Cu, but a significant interaction between pH and O₃ on Mn, Pb and Cd was observed. Due to the poor drainage capacity of this soil, leaching of heavy metals was not observed.