Ozone and urban forests in Italy

Ozone levels along urban-to-rural gradients in three Italian cities (Milan, Florence, Bari) showed that average AOT40 values at rural and suburban sites were 2.6 times higher than those determined at urban sites. However, O₃ also exceeded the European criteria to protect forest health at urban sites, even when the standards for human health protection were met. For protecting street trees in Mediterranean cities, the objectives of measurement at urban sites should extend from the protection of human health to the protection of vegetation as well. A review of forest effects on O₃ pollution and of O₃ pollution on forest conditions in Italian cities showed that it was not possible to distinguish the effect of O₃ in the complex mixture of urban pollutants and stressors. A preliminary list of tree species for urban planning in the Mediterranean area shows the average tree capacity of O₃ removal and VOC emission.     

Crops' responses to ozone in Mediterranean environments

The Mediterranean environment, and most of the Italian peninsula, presents some peculiarities in terms of crop response to O₃ since most physiological mechanisms activated upon O₃ exposure, such as stomatal closure, often overlap and interact with those that underlie plant adaptation to drought and hyperosmotic stress, which are typical of these environments. OTC and EDU experiments have demonstrated that O₃ causes strong yield losses when crops are grown without water limitations. However, exposure to water or saline stress significantly reduced O₃ effects on crop yield. In this review, we present the methodological approaches that have been used to study plant-ozone interactions in Italy as well as biochemical, physiological and agronomic responses for representative cropping systems of the Mediterranean climate.     

Ethylenediurea (EDU): a research tool for assessment and verification of the effects of ground level ozone on plants under natural conditions

Ethylenediurea (EDU) has been widely used to prevent ozone (O₃) injury and crop losses in crop plants and growth reductions in forest trees. Successful use requires establishing a dose/response curve for EDU and the proposed plant in the absence of O₃ and in the presence of O₃ before initiating multiple applications to prevent O₃ injury. EDU can be used to verify foliar O₃ symptoms in the field, and to screen plants for sensitivity to O₃ under ambient conditions. Despite considerable research, the mode of action of EDU remains elusive. Additional research on the mode of action of EDU in suppressing O₃ injury in plants may also be helpful in understanding the mode of action of O₃ in causing injury in plants.     

Protection of plants from ambient ozone by applications of ethylenediurea (EDU): A meta-analytic review

A meta-analysis was conducted to quantitatively assess the effects of ethylenediurea (EDU) on ozone (O₃) injury, growth, physiology and productivity of plants grown in ambient air conditions. Results indicated that EDU significantly reduced O₃-caused visible injury by 76%, and increased photosynthetic rate by 8%, above-ground biomass by 7% and crop yield by 15% in comparison with non-EDU treated plants, suggesting that ozone reduces growth and yield under current ambient conditions. EDU significantly ameliorated the biomass and yield of crops and grasses, but had no significant effect on tree growth with an exception of stem diameter. EDU applied as a soil drench at a concentration of 200-400 mg/L has the highest positive effect on crops grown in the field. Long-term research on full-grown tree species is needed. In conclusion, EDU is a powerful tool for assessing effects of ambient [O₃] on vegetation.     

Use of the antiozonant ethylenediurea (EDU) in Italy: verification of the effects of ambient ozone on crop plants and trees and investigation of EDU's mode of action

Twenty-four experiments where EDU was used to protect plants from ozone (O₃) in Italy are reviewed. Doses of 150 and 450 ppm EDU at 2-3 week intervals were successfully applied to alleviate O₃-caused visible injury and growth reductions in crop and forest species respectively. EDU was mainly applied as soil drench to crops and by stem injection or infusion into trees. Visible injury was delayed and reduced but not completely. In investigations on mode of action, EDU was quickly (<2 h) uptaken and translocated to the leaf apoplast where it persisted long (>8 days), as it cannot move via phloem. EDU did not enter cells, suggesting it does not directly affect cell metabolism. EDU delayed senescence, did not affect photosynthesis and foliar nitrogen content, and stimulated antioxidant responses to O₃ exposure. Preliminary results suggest developing an effective soil application method for forest trees is warranted.     

Modelling annual pasture dynamics: Application to stomatal ozone deposition

Modelling ozone (O₃) deposition for impact risk assessment is still poorly developed for herbaceous vegetation, particularly for Mediterranean annual pastures. High inter-annual climatic variability in the Mediterranean area makes it difficult to develop models characterizing gas exchange behaviour and air pollutant absorption suitable for risk assessment. This paper presents a new model to estimate stomatal conductance (g_s) of Trifolium subterraneum, a characteristic species of dehesa pastures. The MEDPAS (MEDiterranean PAStures) model couples 3 modules estimating soil water content (SWC), vegetation growth and g_s. The g_s module is a reparameterized version of the stomatal component of the EMEP DO₃SE O₃ deposition model. The MEDPAS model was applied to two contrasting years representing typical dry and humid springs respectively and with different O₃ exposures. The MEDPAS model reproduced realistically the g_s seasonal and inter-annual variations observed in the field. SWC was identified as the major driver of differences across years. Despite the higher O₃ exposure in the dry year, meteorological conditions favoured 2.1 times higher g_s and 56 day longer growing season in the humid year compared to the dry year. This resulted in higher ozone fluxes absorbed by T. subterraneum in the humid year. High inter-family variability was found in gas exchange rates, therefore limiting the relevance of single species O₃ deposition flux modelling for dehesa pastures. Stomatal conductance dynamics at the canopy level need to be considered for more accurate O₃ flux modelling for present and future climate scenarios in the Mediterranean area.     

Are Bavarian Forests (southern Germany) at risk from ground-level ozone? Assessment using exposure and flux based ozone indices

Exposure and flux-based indices of O₃ risk were compared, at 19 forest locations across Bavaria in southern Germany from 2002 to 2005; leaf symptoms on mature beech trees found at these locations were also examined for O₃ injury. O₃ flux modelling was performed using continuously recorded O₃ concentrations in combination with meteorological and soil moisture data collected from Level II forest sites. O₃ measurements at nearby rural open-field sites proved appropriate as surrogates in cases where O₃ data were lacking at forest sites (with altitude-dependent average differences of about 10% between O₃ concentrations). Operational thresholds of biomass loss for both O₃ indices were exceeded at the majority of the forest locations, suggesting similar risk under long-term average climate conditions. However, exposure-based indices estimated higher O₃ risk during dry years as compared to the flux-based approach. In comparison, minor O₃-like leaf injury symptoms were detected only at a few of the forest sites investigated. Relationships between flux-based risk thresholds and tree response need to be established for mature forest stands for validation of predicted growth reductions under the prevailing O₃ regimes.     

Pretreatment with nitrogen dioxide modifies plant response to ozone

Plant growth inhibition by ozone is significantly affected by previous exposure to nitrogen dioxide. Experiments on the early growth of four crop species showed that daily pretreatment with NO₂ (0.08–0.10 ppm for 3 h) immediately prior to exposure to O₃ (0.08–0.10 ppm for 6 h) increased the inhibition of radish and wheat growth, decreased the inhibition of bush bean growth, but had no effect on the growth of mint. The magnitudes of the interactive effects indicate that in regions where relatively high concentrations of O₃ are produced by photochemical processes, for example, downwind from urban centres, assessments of the impact of O₃ on vegetation based on knowledge of response to O₃ alone may be seriously flawed.     

Advances in understanding ozone impact on forest trees: Messages from novel phytotron and free-air fumigation studies

Recent evidence from novel phytotron and free-air ozone (O₃) fumigation experiments in Europe and America on forest tree species is highlighted in relation to previous chamber studies. Differences in O₃ sensitivity between pioneer and climax species are examined and viewed for trees growing at the harsh alpine timberline ecotone. As O₃ apparently counteracts positive effects of elevated CO₂ and mitigates productivity increases, response is governed by genotype, competitors, and ontogeny rather than species per se. Complexity in O₃ responsiveness increased under the influence of pathogens and herbivores. The new evidence does not conflict in principle with previous findings that, however, pointed to a low ecological significance. This new knowledge on trees' O₃ responsiveness beyond the juvenile stage in plantations and forests nevertheless implies limited predictability due to complexity in biotic and abiotic interactions. Unravelling underlying mechanisms is mandatory for assessing O₃ risks as an important component of climate change scenarios.     

An Air Quality Data Analysis System for Interrelating Effects,Standards, and Needed Source Reductions: Part 8. An Effective Mean O3 Crop Reduction Mathematical Model

A plant injury mathematical model, applied previously to acute and chronic leaf injury data, is used here to model National Crop Loss Assessment Network (NCLAN) data for 15 cultivars and to calculate species parameters from the cultivar analyses. Percent crop yield reduction is estimated as a function of a new parameter, the effective mean O₃ concentration: m_e = [(Σ c_h ^?1/v)/n]^?v, where c_h is the hourly average ambient O₃ concentration for each daytime hour (defined here as 9:00 A.M.–4:00 P.M., always standard time) of data available at an air sampling site for summer (defined here as June 1–August 31), n is the total number of such available hours, and v is an exposure time-concentration parameter, calculated here to be approximately –0.376. Crop yield reduction for soybean is calculated here as z = 0.478 In (tm_e ^2-66) – 0.42, where z is the Gaussian transform of percent crop reduction, t is the hours of exposure (525 h is used here; 7 h/day for 75 days), and In indicates that the natural logarithm is taken of the quantity within parentheses. Crop yield reductions for seven plant species are estimated with similar equations for each of the 1824 site-years of 1981–1983 hourly O₃ concentration data available in the National Aerometric Data Bank (NADB). County-average effective mean O₃ concentrations are indicated by shading on a U.S. map. State-average O₃ parameters and estimated percent crop yield reductions are tabulated. The National Ambient Air Quality Standard (NAAQS) for O₃ specifies that, on the average, the second highest daily maximum 1-h average O₃ concentration in a year shall not exceed 0.12 ppm. For years 1981-1983,71% of the NADB sites recorded annual second highest daily maximum 1-h average O3 concentrations below 0.125 ppm (for summer daytime hours). Ambient O₃ concentrations reduced the total U.S. crop yield an estimated 5% for years 1981–1983. (Summer, daytime, and all acronyms are always used herein as defined above.)     

Assessment of growth and yield losses in two Zea mays L. cultivars (quality protein maize and nonquality protein maize) under projected levels of ozone

Rapid industrialization and economic developments have increased the tropospheric ozone (O₃) budget since preindustrial times, and presently, it is supposed to be a major threat to crop productivity. Maize (Zea mays L.), a C4 plant is the third most important staple crop at global level with a great deal of economic importance. The present study was conducted to evaluate the performance of two maize cultivars [HQPM1: quality protein maize (QPM)] and [DHM117: nonquality protein maize (NQPM)] to variable O₃ doses. Experimental setup included filtered chambers, nonfiltered chambers (NFC), and two elevated doses of O₃ viz. NFC+15 ppb O₃ (NFC+15) and NFC+30 ppb O₃ (NFC+30). During initial growth period, both QPM and NQPM plants showed hormetic effect that is beneficial due to exposure of low doses of a toxicant (NFC and NFC+15 ppb O₃), but at later stages, growth attributes were negatively affected by O₃. Growth indices showed the variable pattern of photosynthate translocation under O₃ stress. Foliar injury in the form of interveinal chlorosis and reddening of leaves due to increased production of anthocyanin pigments was observed at higher concentrations of O₃. One-dimensional gel electrophoresis of leaves taken from NFC+30 showed reductions of major photosynthetic proteins, and differential response was observed between the two test cultivars. Decline in the number of male flowers at elevated O₃ doses suggested damaging effect of O₃ on reproductive structures which might be a cause of productivity losses. Variable carbon allocation pattern particularly to husk leaves, foliar injury, and damage of photosynthetic proteins led to significant reductions in economic yield at higher O₃ doses. PCA showed that both the cultivars responded more or less similarly to O₃ stress in their respective groupings of growth and yield parameters, but magnitude of their response was variable. It is further supported by difference in the significance of correlations between variables of yield and AOT40. Cultivar response reflects that QPM performed better than NQPM against elevated O₃.     

Effect of ambient-level gas-phase peroxides on foliar injury, growth, and net photosynthesis in Japanese radish (Raphanus sativus)

To investigate the effects of ambient-level gas-phase peroxides concurrent with O₃ on foliar injury, photosynthesis, and biomass in herbaceous plants, we exposed Japanese radish (Raphanus sativus) to clean air, 50 ppb O₃, 100 ppb O₃, and 2-3 ppb peroxides + 50 ppb O₃ in outdoor chambers. Compared with exposure to 100 ppb O₃, exposure to 2-3 ppb peroxides + 50 ppb O₃ induced greater damage in foliar injury, net photosynthetic rates and biomass; the pattern of foliar injury and the cause of net photosynthetic rate reduction also differed from those occurring with O₃ exposure alone. These results indicate for the first time that sub-ppb peroxides + 50 ppb O₃ can cause more severe damage to plants than 100 ppb O₃, and that not only O₃, but also peroxides, could be contributing to the herbaceous plant damage and forest decline observed in Japan's air-polluted urban and remote mountains areas.     

Volatile organic compounds from Italian vegetation and their interaction with ozone

Volatile Organic Compounds (VOCs) emitted from vegetation (particularly isoprenoids) represent an important source of atmospheric hydrocarbons almost double the anthropogenic source. When biogenic VOC mix with NO_x in the presence of UV radiation, ozone (O₃) is formed. In Italy, optimal conditions for O₃ formation in terms of VOC/NO_x ratios and abundance of UV radiation occur for long periods of the year. Moreover, Italian vegetation includes several species that are strong and evergreen isoprenoid emitters, and high temperatures for part of the year further stimulate these temperature-dependent emissions. We review emission of isoprenoids from Italian vegetation, current knowledge on the impact of rising O₃ levels on isoprenoid emission, and evidence showing that isoprenoids can increase both the O₃ flux to the plant and protection against oxidative stress because of their antioxidant functions. This trait not only influences plant tolerance to O₃ but also may substantially alter the flux of O₃ between atmosphere and biosphere.     

An Air Quality Data Analysis System for Interrelating Effects,Standards, and Needed Source Reductions: Part 10. Potential Ambient O3 Standards to Limit Soybean Crop Reduction

Soybean percent crop reduction was estimated as a function of ambient O₃ concentrations for each of 80 agricultural sites in the National Aerometric Data Bank (NADB) for each available year of data for years 1981-1985. Fourteen O₃ concentration statistics were calculated for each of the resulting 320 site-years of data. The two statistics that correlated best with estimated crop reduction were an effective mean O₃ concentration (1 percent of variance unexplained) and an arithmetic mean O₃ concentration (4 percent unexplained). The worst correlation of the 14 was for the statistic used in the present O₃ National Ambient Air Quality Standard (NAAQS), the second highest daily maximum 1-h O₃ concentration (42 percent unexplained). The number of site-years for estimated percent soybean yield reductions was plotted versus increasing O₃ concentrations for each of the 14 O₃ statistics. A maximum crop reduction line was drawn on each plot. These lines were used to estimate (and list) potential ambient O₃ standards for each of the 14 statistics that would limit soybean crop reduction at agricultural. NADB sites to 5, 10, 15, or 20 percent.     

Leaf age affects the responses of foliar injury and gas exchange to tropospheric ozone in Prunus serotina seedlings

We investigated the effect of leaf age on the response of net photosynthesis (A), stomatal conductance (g_wv), foliar injury, and leaf nitrogen concentration (N_L) to tropospheric ozone (O₃) on Prunus serotina seedlings grown in open-plots (AA) and open-top chambers, supplied with either carbon-filtered or non-filtered air. We found significant variation in A, g_wv, foliar injury, and N_L (P < 0.05) among O₃ treatments. Seedlings in AA showed the highest A and g_wv due to relatively low vapor pressure deficit (VPD). Older leaves showed significantly lower A, g_wv, N_L, and higher foliar injury (P < 0.001) than younger leaves. Leaf age affected the response of A, g_wv, and foliar injury to O₃. Both VPD and N_L had a strong influence on leaf gas exchange. Foliar O₃-induced injury appeared when cumulative O₃ uptake reached 8-12 mmol m⁻², depending on soil water availability. The mechanistic assessment of O₃-induced injury is a valuable approach for a biologically relevant O₃ risk assessment for forest trees.     

A comparison of North American and Asian exposure–response data for ozone effects on crop yields

Modelling-based studies to assess the extent and magnitude of ozone (O₃) risk to agriculture in Asia suggest that yield losses of 5–20% for important crops may be common in areas experiencing elevated O₃ concentrations. These assessments have relied on European and North American dose–response relationships and hence assumed an equivalent Asian crop response to O₃ for local cultivars, pollutant conditions and climate. To test this assumption we collated comparable dose–response data derived from fumigation, filtration and EDU experiments conducted in Asia on wheat, rice and leguminous crop species. These data are pooled and compared with equivalent North American dose–response relationships. The Asian data show that at ambient O₃ concentrations found at the study sites (which vary between ～35–75 ppb 4–8 h growing season mean), yield losses for wheat, rice and legumes range between 5–48, 3–47 and 10–65%, respectively. The results indicate that Asian grown wheat and rice cultivars are more sensitive to O₃ than the North American dose–response relationships would suggest. For legumes the scatter in the data makes it difficult to reach any equivalent conclusion in relative sensitivities. As such, existing modelling-based risk assessments may have substantially underestimated the scale of the problem in Asia through use of North American derived dose–response relationships.     

Temporal processes that contribute to nonlinearity in vegetation responses to ozone exposure and dose

Ozone interacts with plant tissue through distinct temporal processes. Sequentially, plants are exposed to ambient O₃ that (1) moves through the leaf boundary layer, (2) is taken up into plant tissue primarily through stomata, and (3) undergoes chemical interaction within plant tissue, first by initiating alterations and then as part of plant detoxification and repair. In this paper, we discuss the linkage of the temporal variability of apoplastic ascorbate with the diurnal variability of defense mechanisms in plants and compare this variability with daily maximum O₃ concentration and diurnal uptake and entry of O₃ into the plant through stomata. We describe the quantitative evidence on temporal variability in concentration and uptake and find that the time incidence for maximum defense does not necessarily match diurnal patterns for maximum O₃ concentration or maximum uptake. We suggest that the observed out-of-phase association of the diurnal patterns for the above three processes produces a nonlinear relationship that results in a greater response from the higher hourly average O₃ concentrations than from the lower or mid-level values. The fact that these out-of-phase processes affect the relationship between O₃ exposure/dose and vegetation effects ultimately impact the ability of flux-based indices to predict vegetation effects accurately for purposes of standard setting and critical levels. Based on the quantitative aspect of temporal variability identified in this paper, we suggest that the inclusion of a diurnal pattern for detoxification in effective flux-based models would improve the predictive characteristics of the models. While much of the current information has been obtained using high O₃ exposures, future research results derived from laboratory biochemical experiments that use short but elevated O₃ exposures should be combined with experimental results that use ambient-type exposures over longer periods of time. It is anticipated that improved understanding will come from future research focused on diurnal variability in plant defense mechanisms and their relationship to the diurnal variability in ambient O₃ concentration and stomatal conductance. This should result in more reliable O₃ exposure standards and critical levels.     

Effects of elevated atmospheric CO2 and tropospheric O3 on tree branch growth and implications for hydrologic budgeting

The forest hydrologic budget may be impacted by increasing CO₂ and tropospheric O₃. Efficient means to quantify such effects are beneficial. We hypothesized that changes in the balance of canopy interception, stem flow, and through-fall in the presence of elevated CO₂ and O₃ could be discerned using image analysis of leafless branches. We compared annual stem flow to the results of a computerized analysis of all branches from the 2002, 2004, and 2006 annual growth whorls of 97 ten-year-old trees from the Aspen Free-Air CO₂ and O₃ Enrichment (Aspen FACE) experiment in Rhinelander, WI. We found significant effects of elevated CO₂ and O₃ on some branch metrics, and that the branch metrics were useful for predicting stem flow from birch, but not aspen. The results of this study should contribute to development of techniques for efficient characterization of effects on the forest hydrologic budget of increasing CO₂ and tropospheric O₃.     

Photosynthesis, chloroplast pigments, and antioxidants in Pinus canariensis under free-air ozone fumigation

High O₃ levels, driving uptake and challenging defense, prevail on the Canary Islands, being associated with the hot and dry summers of the Mediterranean-type climate. Pinus canariensis is an endemic conifer species that forms forests across these islands. We investigated the effects of ozone on photosynthesis and biochemical parameters of P. canariensis seedlings exposed to free-air O₃ fumigation at Kranzberg Forest, Germany, where ambient O₃ levels were similar to those at forest sites in the Canary Islands. The twice-ambient O₃ regime (2xO₃) neither caused visible injury-like chlorotic or necrotic spots in the needles nor significantly affected violaxanthin, antheraxanthin and zeaxanthin levels and the de-epoxidation state of the xanthophyll cycle. In parallel, stomatal conductance for water vapour, net photosynthesis, intercellular CO₂ concentration, chlorophyll fluorescence parameters, as well as antioxidant levels were hardly affected. It is concluded that presently prevailing O₃ levels do not impose severe stress on P. canariensis seedlings.