Ozone (O3) critical levels (CLe) are still poorly developed for herbaceous vegetation. They are currently based on single species responses which do not reflect the multi-species nature of semi-natural vegetation communities. Also, the potential effects of other factors like the nitrogen (N) input are not considered in their derivation, making their use uncertain under natural conditions.Exposure- and dose-response relationships were derived from two open-top chamber experiments exposing a mixture of 6 representative annual Mediterranean pasture species growing in natural soil to 4 O3 fumigation levels and 3 N inputs. The Deposition of O3 and Stomatal Exchange model (DO3SE) was modified to account for the multi-species nature of the canopy following a big-leaf approach. This new approach was used for estimating a multi-species phytotoxic O3 dose (PODy-MS). Response relationships were derived based on O3 exposure (AOT40) and flux (PODy-MS) indices.The treatment effects were similar in the two seasons: O3 reduced the aboveground biomass growth and N modulated this response. Gas exchange rates presented a high inter-specific variability and important inter-annual fluctuations as a result of varying growing conditions during the two years. The AOT40-based relationships were not statistically significant except when the highest N input was considered alone. In contrast, PODy-MS relationships were all significant but for the lowest N input level. The influence of the N input on the exposure- and dose-response relationships implies that N can modify the O3 CLe. However, this is an aspect that has not been considered so far in the methodologies for establishing O3 CLe. Averaging across N input levels, a multi-species O3 CLe (CLef-MS) is proposed POD1-MS = 7.9 mmol m⁻², accumulated over 1.5 month with a 95% confidence interval of (5.9, 9.8). Further efforts will be needed for comparing the CLef-MS with current O3 CLef based on single species responses.

In a free-air fumigation experiment with subalpine grassland, we studied long-term effects of elevated ozone (O3) and nitrogen (N) deposition on ecosystem N pools and on the fate of anthropogenic N. At three times during the seventh year of exposure, N pools and recovery of a stable isotope tracer (15N) were determined in above- and belowground plant parts, and in the soil. Plants were much better competitors for 15N than soil microorganisms. Plant N pools increased by 30–40% after N addition, while soil pools remained unaffected, suggesting that most of the extra N was taken up and stored in plant biomass, thus preventing the ecosystem from acquiring characteristics of eutrophication. Elevated O3 caused an increase of N in microbial biomass and in stabilized soil N, probably resulting from increased litter input and lower litter quality. Different from individual effects, the interaction between the pollutants remained partly unexplained.

Stomatal ozone flux and flux–response relationships were derived for winter wheat (Triticum aestivum L.) grown under fully open-air ozone fumigation. A stomatal conductance (gₛₜₒ) model developed for wheat in Europe was re-parameterized. Compared to European model parameterizations, the main changes were that the VPD and radiation response functions were made less and more restrictive, respectively, and that the temperature function was omitted. The re-parameterized gₛₜₒ model performed well with an r² value of 0.76. The slope and intercept of the regression between observed and predicted gₛₜₒ were not significantly different from 1 to 0, respectively. An ozone uptake threshold of 12 nmol m⁻² s⁻¹ was judged most reasonable for the wheat flux–response relationship in subtropical China. Judging from both flux- and concentration-based relationships, the cultivars investigated seem to be more sensitive to ozone than European cultivars. The new flux–response relationship can be applied to ozone risk assessment in subtropical regions.

The hypothesis was tested that O₃-induced changes in leaf-level photosynthetic parameters have the capacity of limiting the seasonal photosynthetic carbon gain of adult beech trees. To this end, canopy-level photosynthetic carbon gain and respiratory carbon loss were assessed in European beech (Fagus sylvatica) by using a physiologically based model, integrating environmental and photosynthetic parameters. The latter were derived from leaves at various canopy positions under the ambient O₃ regime, as prevailing at the forest site (control), or under an experimental twice-ambient O₃ regime (elevated O₃), as released through a free-air canopy O₃ fumigation system. Gross carbon gain at the canopy-level declined by 1.7%, while respiratory carbon loss increased by 4.6% under elevated O₃. As this outcome only partly accounts for the decline in stem growth, O₃-induced changes in allocation are referred to and discussed as crucial in quantitatively linking carbon gain with stem growth.

Air pollution is frequently proposed as a cause of the increased incidence of allergy in industrialised countries. We investigated the impact of ozone (O₃) on reactive oxygen species (ROS) and allergen content of ragweed pollen (Ambrosia artemisiifolia). Pollen was exposed to acute O₃ fumigation, with analysis of pollen viability, ROS and nitric oxide (NO) content, activity of nicotinamide adenine dinucleotide phosphate (NAD[P]H) oxidase, and expression of major allergens. There was decreased pollen viability after O₃ fumigation, which indicates damage to the pollen membrane system, although the ROS and NO contents were not changed or were only slightly induced, respectively. Ozone exposure induced a significant enhancement of the ROS-generating enzyme NAD(P)H oxidase. The expression of the allergen Amb a 1 was not affected by O₃, determined from the mRNA levels of the major allergens. We conclude that O₃ can increase ragweed pollen allergenicity through stimulation of ROS-generating NAD(P)H oxidase.

In 2006, a controlled infection study was performed in the ‘Kranzberger Forst’ to address the following questions: (1) Will massive artificial inoculation with Apiognomonia errabunda override the previously observed inhibitory effect of chronic ozone? (2) Can biochemical or molecular markers be detected to account for the action of ozone? To this end six adult beech trees were chosen, three ozone fumigated (2× ozone) and three control trees (ambient = 1× ozone). Spore-sprayed branches of sun and shade crown positions of each of the trees, and uninoculated control branches, were enclosed in 100-L plastic bags for one night to facilitate infection initiation. Samples were taken within a five-week period after inoculation. A. errabunda infestation levels quantified by real-time PCR increased in leaves that were not fumigated with additional ozone. Cell wall components and ACC (ethylene precursor 1-amino cyclopropane-1-carboxylic acid) increased upon ozone fumigation and may in part lead to the repression of fungal infection. Chronic sublethal ozone exposure reduces both natural and artificial infestation of beech leaves by the endophytic fungus Apiognomonia errabunda.

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.

Common air pollutants, such as nitrogen oxides (NOₓ), emitted in diesel exhaust, and ozone (O₃), have been implicated in the decline of pollinating insects. Reductionist laboratory assays, focused upon interactions between a narrow range of flowering plant and pollinator species, in combination with atmospheric chemistry models, indicate that such pollutants can chemically alter floral odors, disrupting the cues that foraging insects use to find and pollinate flowers. However, odor environments in nature are highly complex and pollination services are commonly provided by suites of insect species, each exhibiting different sensitivities to different floral odors. Therefore, the potential impacts of pollution-induced foraging disruption on both insect ecology, and the pollination services that insects provide, are currently unknown. We conducted in-situ field studies to investigate whether such pollutants could reduce pollinator foraging and as a result the pollination ecosystem service that those insects provide. Using free-air fumigation, we show that elevating diesel exhaust and O₃, individually and in combination, to levels lower than is considered safe under current air quality standards, significantly reduced counts of locally-occurring wild and managed insect pollinators by 62–70% and their flower visits by 83–90%. These reductions were driven by changes in specific pollinator groups, including bees, flies, moths and butterflies, and coincided with significant reductions (14–31%) in three different metrics of pollination and yield of a self-fertile test plant. Quantifying such effects provides new insights into the impacts of human-induced air pollution on the natural ecosystem services upon which we depend.

Frequency and intensity of wildfire occurrences are dramatically increasing worldwide due to global climate change, having a devastating effect on the entire ecosystem including plants. Moreover, distribution of fire-smoke can influence the natural environment over very long distances, i.e. hundreds of kilometres. Dry plant matter contains 0.1–0.9% (w/w) sulphur, which is mainly released during combustion into the atmosphere as sulphur dioxide (SO₂) resulting in local concentrations of up to 3000 nL L⁻¹. SO₂ is a highly hazardous gas, which enters plants mostly via the stomata. Toxic sulphite is formed inside the leaves due to conversion of SO₂. Plants as sessile organisms cannot escape from threats, why they evolved an impressive diversity of molecular defence mechanisms. In the present study, two recent wildfires in Germany were evaluated to analyse the effect of SO₂ released into the atmosphere on deciduous trees: the Meppen peat fire in 2018 and the forest fire close to Luebtheen in 2019. Collected leaf material from beech (Fagus sylvatica) and oak (Quercus robur) was examined with respect to detoxification of sulphur surplus due to the exposure to elevated SO₂. An induced stress reaction in both species was indicated by a 1.5-fold increase in oxidized glutathione. In beech leaves, the enzymatic activities of the sulphite detoxification enzymes sulphite oxidase and apoplastic peroxidases were increased 5-fold and a trend of sulphate accumulation was observed. In contrast, oaks did not regulate these enzymes during smoke exposure, however, the constitutive activity is 10-fold and 3-fold higher than in beech. These results show for the first time sulphite detoxification strategies of trees in situ after natural smoke exposure. Beech and oak trees survived short-term SO₂ fumigation due to exclusion of toxic gases and different oxidative detoxification strategies. Beeches use efficient upregulation of oxidative sulphite detoxification enzymes, while oaks hold a constitutively high enzyme-pool available.