Ozone (O3) is a photochemically formed reactive gas responsible for a decreasing carbon assimilation in plant ecosystems. Present in the atmosphere in trace concentrations (less than 100 ppbv), this molecule is capable of inhibiting carbon assimilation in agricultural and forest ecosystems. Ozone-risk assessments are typically based on manipulative experiments. Present regulations regarding critical ozone levels are mostly based on an estimated accumulated exposure over a given threshold concentration. There is however a scientific consensus over flux estimates being more accurate, because they include plant physiology analyses and different environmental parameters that control the uptake—that is, not just the exposure—of O3. While O3 is a lot more difficult to measure than other non-reactive greenhouse gases, UV-based and chemiluminescence sensors enable precise and fast measurements and are therefore highly desirable for eddy covariance studies. Using micrometeorological techniques in association with latent heat flux measurements in the field allows for the partition of ozone fluxes into the stomatal and non-stomatal sinks along the soil-plant continuum. Long-term eddy covariance measurements represent a key opportunity in estimating carbon assimilation at high-temporal resolutions, in an effort to study the effect of climate change on photosynthetic mechanisms. Our aim in this work is to describe potential of O3 flux measurement at the canopy level for ozone-risk assessment in established long-term monitoring networks.

Aminocyclopyrachlor is a pyrimidine carboxylic acid herbicide used to control broadleaf weeds and brush. Amending soil with activated charcoal is recommended to prevent off-site transport of aminocyclopyrachlor and non-target plant damage. We used the batch-equilibrium method to determine the concentration of aminocyclopyrachlor in a pseudo-steady state with biochar, soil, and biochar-soil systems (<10% biochar by weight). We observed that aminocyclopyrachlor is mobile in soils. Soil incorporation of activated charcoal removed nearly all of the aqueous aminocyclopyrachlor thereby limiting its bioavailability to non-target flora. On the other hand, biochars were less effective than activated charcoal. Biochar produced from olive mill waste feedstock was the most effective biochar that we assessed for reducing the aqueous herbicide concentration. Although these biochars reduced the aminocyclopyrachlor concentration, they would not be practical remediation media due to the extraordinarily high application rates required to reduce the concentration by 50% (2.13 × 105 kg ha−1–7.27 × 105 kg ha−1).

An account of histo-cytological and ultrastructural studies on ozone effect on crop and forest species in Italy is given, with emphasis on induced cell death and the underlying mechanisms. Cell death phenomena possibly due to ambient O3 were recorded in crop and forest species. In contrast, visible O3 effects on Mediterranean vegetation are often unclear. Microscopy is thus suggested as an effective tool to validate and evaluate O3 injury to Mediterranean vegetation. A DAB-Evans blue staining was proposed to validate O3 symptoms at the microscopic level and for a pre-visual diagnosis of O3 injury. The method has been positively tested in some of the most important crop species, such as wheat, tomato, bean and onion and, with some restriction, in forest species, and it also allows one to gain some very useful insights into the mechanisms at the base of O3 sensitivity or tolerance. Ozone-induced cell death is a frequent phenomenon in Mediterranean conditions, not only in the most sensitive crops but also in forest species.

The negative health effects of simulated acid mists and fluoranthene on juvenile Japanese red pine were investigated, and the methods of protection from these pollutants were examined. The needle gas exchange, chlorophyll fluorescence, chemical contents and visual damage to needles caused by acid mist applied alone or its conjunction with fluoranthene were investigated over 60 d and 20 d, respectively. Acid mist at pH 2 and 3 caused physiological and visual damage, which was enhanced by the addition of fluoranthene to the mist. However, fluoranthene and acid mist at pH 4 and 5 showed only minor effects. These findings indicate that acid mist may be more harmful to pine trees if it occurs in conjunction with polycyclic aromatic hydrocarbons. Moreover, suppression of the singular and additive effects of these compounds was achieved using mannitol, which may be widely applicable to suppression of reactive oxygen species-mediated plant damage.

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.

Plants react to O3 threat by setting up a variety of defensive strategies involving the co-ordinated modulation of stress perception, signalling and metabolic responses. Although stomata largely controls O3 uptake, differences in O3 tolerance cannot always be ascribed to changes in stomatal conductance but cell protective and repair processes should be taken into account. O3-driven ROS production in the apoplast induces a secondary, active, self-propagating generation of ROS, whose levels must be finely tuned, by many enzymatic and non-enzymatic antioxidant systems, to induce gene activation without determining uncontrolled cell death. Additional signalling molecules, as ethylene, jasmonic and salicylic acid are also crucial to determine the spreading and the containment of leaf lesions. The main recent results obtained on O3 sensing, signal transduction, ROS formation and detoxification mechanisms are here discussed. A dissection of the complex network of interacting mechanisms which determine the cell fate under ozone stress.

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. EDU was successful as a tool in determining ozone effects on vegetation in Italy, but while progress was made, its mode of action remains unresolved.

An Open-Top Chambers experiment on Fagus sylvatica and Quercus robur seedlings was conducted in order to compare the performance of an exposure-based (AOT40) and a flux-based approaches in predicting the appearance of ozone visible injuries on leaves. Three different ozone treatments (charcoal-filtered; non-filtered; and open plots) and two soil moisture treatments (watered and non-watered plots) were performed. A Jarvisian stomatal conductance model was drawn up and parameterised for both species and typical South Alpine environmental conditions, thus allowing the calculation of ozone stomatal fluxes for every treatment. A critical ozone flux level for the onset of leaf visible injury in beech was clearly identified between 32.6 and 33.6 mmol O3m-2. In contrast, it was not possible to identify an exposure critical level using the AOT40 index. Water stress delayed the onset of the leaf visible injuries, but the flux-based approach was able to take it into account accurately. Ozone fluxes are more accurate than AOT40 exposure index in predicting ozone visible foliar injury onset on beech seedlings in South Alpine environmental conditions.

Nitrate and ammonium concentration in wet deposition detrimentally impacted a sensitive pollution indicator species irrespective of the nitrogen dose.

Soybean [Glycine max (L.) Merr.] cultivars Essex and Forrest that exhibit differences in ozone (O3) sensitivity were used in greenhouse experiments to investigate the role of leaf extracellular antioxidants in O3 injury responses. Charcoal-filtered air and elevated O3 conditions were used to assess genetic, leaf age, and O3 effects. In both cultivars, the extracellular ascorbate pool consisted of 80e98% dehydroascorbic acid, the oxidized form of ascorbic acid (AA) that is not an antioxidant. For all combinations of genotype and O3 treatments, extracellular AA levels were low (1e30 nmol g 1 FW) and represented 3e30% of the total antioxidant capacity. Total extracellular antioxidant capacity was twofold greater in Essex compared with Forrest, consistent with greater O3 tolerance of Essex. The results suggest that extracellular antioxidant metabolites in addition to ascorbate contribute to detoxification of O3 in soybean leaves and possibly affect plant sensitivity to O3 injury.