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.

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.

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.

To investigate the interactive effects of increasing [CO₂] and heat wave occurrence on isoprene (IE) and methanol (ME) emissions, Platanus orientalis was grown for one month in ambient (380 mmol mol 1) or elevated (800 mmol mol 1) [CO₂] and exposed to high temperature (HT) (38 C/4 h). In pre-existing leaves, IE emissions were always higher but ME emissions lower as compared to newly-emerged leaves. They were both stimulated by HT. Elevated [CO₂] significantly reduced IE in both leaf types, whereas it increased ME in newly-emerged leaves only. In newly-emerged leaves, elevated [CO₂] decreased photosynthesis and altered the chloroplast ultrastructure and membrane integrity. These harmful effects were amplified by HT. HT did not cause any unfavorable effects in pre-existing leaves, which were characterized by inherently higher IE rates. We conclude that: (1) these results further prove the isoprene's putative thermo-protective role of membranes; (2) HT may likely outweigh the inhibitory effects of elevated [CO₂] on IE in the future.

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.

A rapid, quantitative bioassay for detecting phytotoxic air pollutants has been developed. The technique uses wheat Triticum aestivum L. or tomato Lycopersicon esculentum L., seedlings growing on an agar medium in test-tubes. The seedlings are exposed to a pollutant in the test-tube and stress-ethylene induced by the pollutant is quantitatively measured by gas chromatography. Increases in ethylene production from seedlings exposed to a phytotoxic air pollutant as compared with controls not exposed to pollutants were related to the pollutant concentration.

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).

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.