In a world with climate change and environmental pollution, modern Biology is concerned with organismic susceptibility. At the same time, policy and decision makers seek information about organismic susceptibility. Therefore, information about organismic susceptibility may have far-reaching implications to the entire biosphere that can extend to several forthcoming generations. Here, we review a sample of approximately 200 published peer-reviewed articles dealing with plant response to ground-level ozone to understand how the information about susceptibility is communicated. A fuzzy and often incorrect terminology was used to describe the responsiveness of plants to ozone. Susceptibility was classified too arbitrarily and this was reflected to the approximately 50 descriptive words that were used to characterize susceptibility. The classification of susceptibility was commonly based on calculated probability (p) value. This practice is inappropriate as p values do not provide any basis for effect or susceptibility magnitude. To bridge the gap between science and policy decision making, classification of susceptibility should be done using alternative approaches, such as effect size estimates in conjunction with multivariate ordination statistics.

Ground-level ozone (O₃) pollution can adversely affect human health and vegetation, thus being an important environmental issue nowadays. Ozone biological monitoring (biomonitoring) is a method of O₃ monitoring by observing quantitative changes in living organisms physically present in a specific environment. Here, we provide a concise view of the field of O₃ biomonitoring, along with recent advances that are expected to advance this field in the future. We also recommend that O₃ biomonitoring is included in citizen science initiatives as well as in worldwide curricula of educational institutions. Policy-makers and general public may not understand biomonitoring data; hence, a major challenge is how to communicate the information to the audience in a way that permits the best comprehension.

Hormesis is a fundamental notion in ecotoxicology while competition between organisms is an essential notion in population ecology and species adaptation and evolution. Both sub-disciplines of ecology deal with the response of organisms to abiotic and biotic stresses. In ecotoxicology, the Linear-non-Threshold (LNT), Threshold and Hormetic models are used to describe the dominant responses of a plethora of endpoints to abiotic stress. In population ecology, the logistic, theta-logistic and the Allee effect models are used to describe the growth of populations under different responses to (biotic) stress induced by population density. The per capita rate of population increase (r) measures species fitness. When it is used as endpoint, the responses to population density seem to perfectly correspond to LNT, Threshold and Hormetic responses to abiotic stress, respectively. Our analysis suggests the Allee effect is a hormetic-like response of r to population density, an ultimate biotic stress. This biphasic dose-response model appears across different systems and situations (from molecules to tumor growth to population dynamics), is highly supported by ecological and evolutionary theory, and has important implications in most sub-disciplines of biology as well as in environmental and earth sciences. Joined multi-disciplinary efforts would facilitate the development and application of advanced research approaches for better understanding potential planetary-scale implications.

The nature of the dose-response relationship in the low dose zone and how this concept may be used by regulatory agencies for science-based policy guidance and risk assessment practices are addressed here by using the effects of surface ozone (O₃) on plants as a key example for dynamic ecosystems sustainability. This paper evaluates the current use of the linear non-threshold (LNT) dose-response model for O₃. The LNT model has been typically applied in limited field studies which measured damage from high exposures, and used to estimate responses to lower concentrations. This risk assessment strategy ignores the possibility of biological acclimation to low doses of stressor agents. The upregulation of adaptive responses by low O₃ concentrations typically yields pleiotropic responses, with some induced endpoints displaying hormetic-like biphasic dose-response relationships. Such observations recognize the need for risk assessment flexibility depending upon the endpoints measured, background responses, as well as possible dose-time compensatory responses. Regulatory modeling strategies would be significantly improved by the adoption of the hormetic dose response as a formal/routine risk assessment option based on its substantial support within the literature, capacity to describe the entire dose-response continuum, documented explanatory dose-dependent mechanisms, and flexibility to default to a threshold feature when background responses preclude application of biphasic dose responses.

Ammonium sulfate [(NH₄)₂SO₄] deposition and elevated ozone (O₃) concentrations may negatively affect plants and trophic interactions. This study aimed to evaluate for the first time the interactive effects of high (NH₄)₂SO₄ load and elevated O₃ levels on cauliflower (Brassica oleracea L.) under field conditions. Cauliflower seedlings were treated with 0 (AS0) or 50 (AS50) kg ha⁻¹ (NH₄)₂SO₄ and exposed to ambient (AOZ, ≈20 ppb) or elevated (EOZ, ≈55 ppb) O₃ for about one month, in a Free Air O₃ Concentration Enrichment (FACE) system. The oligophagous diamondback moth (Plutella xylostella Linnaeus, 1758) showed a clear preference towards the seedlings treated with AS50, which intensively grazed. Plant-herbivore interactions were driven by (NH₄)₂SO₄ availability, rather than O₃, via increased nitrogen content in the leaves. Further laboratory bioassays were followed to confirm the validity of these observations using polyphagous Eri silkmoth larvae (Samia ricini) as a biological model in a standardized experimental setup. Choice assays, where larvae could select leaves among leaf samples from the different experimental conditions, and no-choice assays, where larvae could graze leaves from just one experimental condition, were conducted. In the choice assay, the larvae preferred AS50-treated leaves, in agreement with the field observations with diamondback moth. In the no-choice assay, larval body mass growth was inhibited when fed with leaves treated with EOZ and/or AS50. Larvae fed with AS50-treated leaves displayed increased mortality. These observations coincide with higher NO₃ and Zn content in AS50-treated leaves. This study shows that plant-herbivore interactions can be driven by (NH₄)₂SO₄ availability, independently of O₃, and suggests that high N deposition may have severe health implications in animals consuming such plant tissues.

Key message: Plant-herbivore interactions are driven by high (NH₄)₂SO₄ availability, independently of O₃.

The United States Environmental Protection Agency (US EPA) has recently proposed changes to strengthen the transparency of its pivotal regulatory science policy and procedures. In this context, the US EPA aims to enhance the transparency of dose-response data and models, proposing to consider for the first time non-linear biphasic dose-response models. While the proposed changes have the potential to lead to markedly improved ecological risk assessment compared to past and current approaches, we believe there remain open issues for improving the quality of ecological risk assessment, such as the consideration of adaptive, dynamic and interactive effects. Improved risk assessment including adaptive and dynamic non-linear models (beyond classic threshold models) can enhance the quality of regulatory decisions and the protection of ecological health. We suggest that other countries consider adopting a similar scientific-regulatory posture with respect to dose-response modeling via the inclusion of non-linear biphasic models, that incorporate the dynamic potential of biological systems to adapt (i.e., enhancing positive biological endpoints) or maladapt to low levels of stressor agents.

Evaluations of ozone effects on vegetation across the globe over the last seven decades have mostly incorporated exposure levels that were multi-fold the preindustrial concentrations. As such, global risk assessments and derivation of critical levels for protecting plants and food supplies were based on extrapolation from high to low exposure levels. These were developed in an era when it was thought that stress biology is framed around a linear dose-response. However, it has recently emerged that stress biology commonly displays non-linear, hormetic processes. The current biological understanding highlights that the strategy of extrapolating from high to low exposure levels may lead to biased estimates. Here, we analyzed a diverse sample of published empirical data of approximately 500 stimulatory, hormetic-like dose-responses induced by ozone in plants. The median value of the maximum stimulatory responses induced by elevated ozone was 124%, and commonly <150%, of the background response (control), independently of species and response variable. The maximum stimulatory response to ozone was similar among types of response variables and major plant species. It was also similar among clades, between herbaceous and woody plants, between deciduous and evergreen trees, and between annual and perennial herbaceous plants. There were modest differences in the stimulatory response between genera and between families which may reflect different experimental designs and conditions among studies. The responses varied significantly upon type of exposure system, with open-top chambers (OTCs) underestimating the maximum stimulatory response compared to free-air ozone-concentration enrichment (FACE) systems. These findings suggest that plants show a generalized hormetic stimulation by ozone which is constrained within certain limits of biological plasticity, being highly generalizable, evolutionarily based, and maintained over ecological scales. They further highlight that non-linear responses should be taken into account when assessing the ozone effects on plants.

Ground-level ozone (O₃) levels are nowadays elevated in wide regions of the Earth, causing significant effects on plants that finally lead to suppressed productivity and yield losses. Ethylenediurea (EDU) is a chemical compound which is widely used in research projects as phytoprotectant against O₃ injury. The EDU mode of action remains still unclear, while there are indications that EDU may contribute to plants with nitrogen (N) when the soil is poor in N and the plants have relatively small leaf area. To reveal whether the N content of EDU acts as a fertilizer to plants when the soil is not poor in N and the plants have relatively large total plant leaf area, willow plants (Salix sachalinensis Fr. Schm) were exposed to low ambient O₃ levels and treated ten times (9-day interval) with 200 mL soil drench containing 0, 800 or 1600 mg EDU L⁻¹. Fertilizer was added to a nutrient-poor soil, and the plants had an average plant leaf area of 9.1 m² at the beginning of EDU treatments. Indications for EDU-induced hormesis in maximum electron transport rate (J_max) and ratio of intercellular to ambient CO₂ concentration (C_i:C_a) were observed at the end of the experiment. No other EDU-induced effects on leaf greenness and N content, maximum quantum yield of photosystem II (F_v/F_m), gas exchange, growth and matter production suggest that EDU did not act as N fertilizer and did not cause toxicity under these experimental conditions.

Ground-level ozone (O₃) concentrations have been elevating in the last century. While there has been a notable progress in understanding O₃ effects on vegetation, O₃ effects on ecological stoichiometry remain unclear, especially early in the oxidative stress. Ethyelenediurea (EDU) is a chemical compound widely applied in research projects as protectant of plants against O₃ injury, however its mode of action remains unclear. To investigate O₃ and EDU effects early in the stress, we sprayed willow (Salix sachalinensis) plants with 0, 200 or 400 mg EDU L⁻¹, and exposed them to either low ambient O₃ (AOZ) or elevated O₃ (EOZ) levels during the daytime, for about one month, in a free air O₃ controlled exposure (FACE); EDU treatment was repeated every nine days. We collected samples for analyses from basal, top, and shed leaves, before leaves develop visible O₃ symptoms. We found that O₃ altered the ecological stoichiometry, including impacts in nutrient resorption efficiency, early in the stress. The relation between P content and Fe content seemed to have a critical role in maintaining homeostasis in an effort to prevent O₃-induced damage. Photosynthetic pigments and P content appeared to play an important role in EDU mode of action. This study provides novel insights on the stress biology which are of ecological and toxicological importance.

Increased mixing ratios of ground-level ozone (O₃) threaten individual plants, plant communities and ecosystems. In this sense, O₃ biomonitoring is of great interest. The O₃-sensitive S156 and the O₃-tolerant R123 genotypes of snap bean (Phaseolus vulgaris L.) have been proposed as a potential tool for active biomonitoring of ambient O₃. In the present study, an O₃ biomonitoring was conducted, with the S156/R123 tool, along with a monitoring of O₃ and other environmental conditions in an urban area in Athens, Greece, during the growing seasons of 2012 and 2013. Plant yield was evaluated to assess the effectiveness of AOT40 in interpreting O₃-induced phytotoxicity. Across the two genotypes, an approximately two times lower total number of pods – and consequently lower bulk mass of seeds – was found in 2012 than in 2013, although there was no significant difference in the final AOT40 between the two years. No significant differences were observed in the stomatal density or conductance between the two genotypes, whereas it was estimated that, in both genotypes, the abaxial leaf surface contributes 2.7 fold to O₃ intake in comparison to the adaxial one. By testing the role of ambient air temperature in outdoor plant environment chambers (OPECs), it was found that increased temperature limits mature pod formation and complicates interpretation of O₃ impacts in terms of S156/R123 yields ratios. This is the first study providing evidence for a hormetic response of plants to ambient air temperature. This study also points out the complexity of using yield as a measure of O₃ impact across different environments with the snap bean system, whereas visible foliar injury is more consistently related to O₃ effects.

The antiozonant and research tool ethylene diurea (EDU) is widely studied as a phytoprotectant against the widespread pollutant ground-surface ozone. Although it has been extensively used, its potential toxicity in the absence of ozone is unknown and its mode of action is unclear. The purpose of this research was to toxicologically assess EDU and to further investigate its mode of action using Lemna minor L. as a model organism. Application of EDU concentrations greater than 593 mg L⁻¹ (practically 600 mg L⁻¹) resulted in adverse inhibition of colony growth. As no-observed-toxic-effects concentration (NOEL) we recommend a concentration of 296 mg L⁻¹ (practically 300 mg L⁻¹). A hormetic response was detected, i.e. stimulatory effects of low EDU concentrations, which may indicate overcompensation in response to disruption in homeostasis. Growth inhibition and suppressed biomass were associated with impacted chlorophyll a fluorescence (Φ_PSII, q_P and ETR). Furthermore, EDU increased mesophyll thickness, as indicated by frond succulence index. Applications of concentrations ≥593 mg L⁻¹ to uncontrolled environments should be avoided due to potential toxicity to sensitive organisms and the environment.