Ozone risk assessment for agricultural crops in Europe: Further development of stomatal flux and flux–response relationships for European wheat and potato

Applications of a parameterised Jarvis-type multiplicative stomatal conductance model with data collated from open-top chamber experiments on field grown wheat and potato were used to derive relationships between relative yield and stomatal ozone uptake. The relationships were based on thirteen experiments from four European countries for wheat and seven experiments from four European countries for potato. The parameterisation of the conductance model was based both on an extensive literature review and primary data. Application of the stomatal conductance models to the open-top chamber experiments resulted in improved linear regressions between relative yield and ozone uptake compared to earlier stomatal conductance models, both for wheat (r²=0.83) and potato (r²=0.76). The improvement was largest for potato. The relationships with the highest correlation were obtained using a stomatal ozone flux threshold. For both wheat and potato the best performing exposure index was AF_st6 (accumulated stomatal flux of ozone above a flux rate threshold of 6 nmol ozone m⁻² projected sunlit leaf area, based on hourly values of ozone flux). The results demonstrate that flux-based models are now sufficiently well calibrated to be used with confidence to predict the effects of ozone on yield loss of major arable crops across Europe. Further studies, using innovations in stomatal conductance modelling and plant exposure experimentation, are needed if these models are to be further improved.     

Modelling stomatal responses of spring wheat (Triticum aestivum L. cv. Turbo) to ozone and different levels of water supply

Spring wheat (Triticum aestivum L. cv. Turbo) was exposed to different levels of ozone and water supply in open-top chambers in 1991. Air was charcoal filtered (CF), non-filtered (NF) and CF plus proportional addition of ambient or twice ambient ozone (CF1, CF2). Seasonal means of O(3), taken over 24 h, were 2.3, 20.6, 17.3, and 34.5 nl litre(-1) for CF, NF, CF1 and CF2 treatments, respectively. A split-plot design was used to obtain two levels of water supply: one-half of the pots was irrigated sufficiently not to show any symptoms of drought stress; the others were exposed to low water supply and received 50% of these amounts. Using a steady-state porometer approximately 800 measurements of stomatal conductance (g(s)) were made on flag leaves from 68 to 106 days after sowing. The measurements yielded only small differences of maximum conductance between the two levels of water supply. Therefore, low water supply did not protect wheat plants against ozone injury via reduced stomatal uptake in this experiment. To describe the effects of environmental variables on the stomatal behaviour, boundary-line analysis and non-linear regression analysis were used. Besides microclimatic parameters, the ozone dose of flag leaves was introduced as an independent variable affecting stomatal aperture. A well-defined boundary line for ozone dose was found, suggesting that increasing ozone dose caused stomatal closure in wheat flag leaves. But at high ozone doses, co-acting senescence seems also responsible for the decrease in stomatal conductance. A multiplicative boundary-line model was used to predict stomatal conductance from combinations of environmental variables. In the test carried out with the measurements of stomatal conductance, the model accounted only for 40% of the variation of g(s). Generalized stomatal response patterns of the herbaceous growth form, the dependence of the variables' age and ozone dose and the lack of an important factor influencing stomatal response (water status of the plant) in the model, are suggested as explanations of the poor results of the test.     

A comparison of two stomatal conductance models for ozone flux modelling using data from two Brassica species

In this study we tested and compared a multiplicative stomatal model and a coupled semi-empirical stomatal-photosynthesis model in their ability to predict stomatal conductance to ozone (g_st) using leaf-level data from oilseed rape (Brassica napus L.) and broccoli (Brassica oleracea L. var. italica Plenck). For oilseed rape, the multiplicative model and the coupled model were able to explain 72% and 73% of the observed g_st variance, respectively. For broccoli, the models were able to explain 53% and 51% of the observed g_st variance, respectively. These results support the coupled semi-empirical stomatal-photosynthesis model as a valid alternative to the multiplicative stomatal model for O₃ flux modelling, in terms of predictive performance.     

Sensitivity analysis of a parameterization of the stomatal component of the DO3SE model for Quercus ilex to estimate ozone fluxes

A sensitivity analysis of a proposed parameterization of the stomatal conductance (g(s)) module of the European ozone deposition model (DO(3)SE) for Quercus ilex was performed. The performance of the model was tested against measured g(s) in the field at three sites in Spain. The best fit of the model was found for those sites, or during those periods, facing no or mild stress conditions, but a worse performance was found under severe drought or temperature stress, mostly occurring at continental sites. The best performance was obtained when both f(phen) and f(SWP) were included. A local parameterization accounting for the lower temperatures recorded in winter and the higher water shortage at the continental sites resulted in a better performance of the model. The overall results indicate that two different parameterizations of the model are needed, one for marine-influenced sites and another one for continental sites.     

Derivation of ozone flux-yield relationships for lettuce: a key horticultural crop

Ozone flux-response relationships were derived for lettuce, employing a multiplicative approach to model the manner in which stomatal conductance is influenced by key environmental variables, using a dataset collected during field experimentation in Crete and yield-response relationships derived from parallel open-top chamber experiments. Regional agronomic practices were adopted throughout. Computed versus measured data revealed that the derived model explained 51% (P<0.001) of the observed variation in stomatal conductance. Concentration-based indices were compared with flux-based indices. Analyses revealed a significant relationship between accumulated stomatal ozone flux and yield employing flux threshold cut-offs up to 4 nmol m(-2) s(-1). Regressions employing very low or zero flux thresholds resulted in the strongest yield-flux relationships (explaining approximately 80% (P<0.05) of the variation in the dataset).     

Canopy profiles of photosynthetic parameters under elevated CO2 and N fertilization in a poplar plantation

A poplar plantation has been exposed to an elevated CO2 concentration for 5 years using the free air CO2 enrichment (FACE) technique. Even after such a long period of exposure, leaves of Populus x euramericana have not shown clear signs of photosynthetic acclimation. Only at the end of the growing season for shade leaves was a decrease of maximum velocity of carboxylation (Vcmax) observed. Maximum electron transport rate (Jmax) was increased by FACE treatment in July. Assimilation rates at CO2 partial pressure of 400 (A400) and 600 (A600) micromol mol(-1) were not significantly different under FACE treatment. Most notably FACE significantly decreased stomatal conductance (g(s)) both on upper and lower canopy leaves. N fertilization increased N content in the leaves on mass basis (Nm) and specific leaf area (SLA) in both CO2 treatments but did not influence the photosynthetic parameters. These data show that in poplar plantations the long-term effects of elevated CO2 on photosynthesis do not differ considerably from the short-term ones even with N deposition.     

Foliage responses of spruce trees to long-term low-grade sulfur dioxide deposition

Foliage on spruce trees (Picea rubens Sarg.) growing on dry SO(2) deposition zones (dry SO(2) deposition ranging from 0.5 and 8.5 S kg ha(-1) year(-1)) downwind from a SO(2) emission source was analyzed to assess chronic effects of long-term low-grade SO(2) deposition on net photosynthesis, stomatal conductance, dark respiration, stomatal antechamber wax structures, elemental concentrations in and on foliage (bulk and surficial concentrations), and types of epiphytic fungi that reside in the phylloplane. Elemental distributions on stomatal antechambers, on fungal colonies, and on smooth surfaces between stomates and fungus colonies were determined with a scanning electronic microscope (SEM) by way of X-ray scanning. It was found that net photosynthesis of newly developed spruce foliage (current-year, and 1-year-old) was not significantly affected by the local SO(2) deposition rates. Sulfur dioxide deposition, however, may have contributed to the gradual decrease in net photosynthesis with increasing needle age. Dark respiration rates were significantly higher on foliage taken from high SO(2) deposition zones. Stomatal rod-web structures deteriorated to flakes with increasing needle age and increasing SO(2) deposition. Further inspection of the needle surfaces revealed an increasing abundance of fungal colonies with increasing needle age. Many fungal taxa were isolated and identified. It was found that black yeasts responded positively, and Xylohypha pinicola responded negatively to high rates of SO(2) deposition. Surficial concentrations of elements such as P, S, K, Cl, Ca were about 10 times higher on fungal colonies than on smooth needle surfaces. Surficial Ca contents on 4 or 5-year-old needles decreased with increasing SO(2) deposition, but surficial S concentrations remained the same. In contrast, bulk foliar Ca and S concentrations increased with increasing SO(2) deposition.     

Does living in elevated CO2 ameliorate tree response to ozone? A review on stomatal responses

Short-term elevated O3 reduces photosynthesis, which reduces stomatal conductance (g(s)) in response to increased substomatal CO2 concentration (Ci). Further exposure causes stomata to become sluggish in response to environmental stimuli. Exposure to elevated CO2 stimulates rapid stomata closure in response to increased Ci. This reduction in g(s) may not be sustained over time as photosynthesis down-regulates and with it, g(s). The relationship between g(s) and photosynthesis may not be constant because stomata respond more slowly to environmental changes than photosynthesis, and because elevated CO2 may alter guard cell sensitivity to other signals. Also, reduced stomatal density (and g(s)) in response to long-term CO2 enrichment suggests sustained reduction in g(s). Elevated CO2 is believed to ameliorate the deleterious O3 effects by reducing g(s) and thus the potential O3 flux into leaves. Confirmation that g(s) acclimation to CO2 enrichment does not lessen over time is critical for developing meaningful O3 flux scenarios.     

Robustness of modelled ozone exposures and doses

This study evaluates the robustness of the AOTX and AF(st)Y indices for assessing the ozone-induced risk to vegetation. These indices represent the accumulated concentration and stomatal flux, respectively, above a threshold value. The robustness is expressed as the sensitivity to changes in inputs and the uncertainty due to input errors. The input data are taken from a regional-scale chemical transport model. Both indices show increasing sensitivity with increasing threshold values. The sensitivity depends on the threshold and the characteristics of the frequency distribution for concentrations and stomatal fluxes. AF(st)Y appears less sensitive than AOTX for the thresholds adopted for critical levels. The couplings between concentration gradients and deposition algorithms complicate the assessment of the total uncertainty. For AF(st)Y, the uncertainty due to the modelled stomatal conductance may sometimes increase, but sometimes decrease, the overall uncertainty significantly. In particular, the maximum stomatal conductance plays an important role in determining the uncertainty.     

Effects of elevated O3 and CO2 on chlorophyll fluorescence and gas exchange in Scots pine during the third growing season

Naturally regenerated, 30-year-old Scots pines (Pinus Sylvestris L.) were grown in open-top chambers and exposed in situ to doubled ambient O(3), doubled ambient CO(2) and a combination of elevated O(3) and CO(2) from 15 April to 15 September for three growing seasons (1994-1996). To examine the effects of O(3) and/or CO(2) on photosynthesis, chlorophyll a fluorescence and gas exchange were measured simultaneously. Doubled ambient O(3) significantly decreased the rates of photosynthesis at all levels of photon flux density. This was related mainly to a significant decrease in the photochemical efficiency of photosystem II (PS II) and the rate of whole electron transport, rather than to a decrease in stomatal conductance. When measurements were made at doubled ambient concentration of CO(2) (700 micromol mol(-1)), doubled ambient CO(2) treatment did not lead to a significant change in the intrinsic capacity of photosynthesis, as manifested by no changes in PS II, the rate of electron transport, the maximal rate of photosynthesis and the apparent quantum yield of CO(2) assimilation. However, elevated CO(2) increased the sensitivity of stomatal conductance to light and decreased maximal stomatal conductance. When O(3) and CO(2) were combined, the O(3)-induced decrease in photosynthesis rate was reduced significantly by a high concentration of CO(2). This may be partly related to the decrease in stomatal conductance induced by the high concentration of CO(2). The complete mechanism behind this interaction is, however, still unclear.     

Implications of climate change for the stomatal flux of ozone: a case study for winter wheat

Climate change factors such as elevated CO2 concentrations, warming and changes in precipitation affect the stomatal flux of ozone (O3) into leaves directly or indirectly by altering the stomatal conductance, atmospheric O3 concentrations, frequency and extent of pollution episodes and length of the growing season. Results of a case study for winter wheat indicate that in a future climate the exceedance of the flux-based critical level of O3 might be reduced across Europe, even when taking into account an increase in tropospheric background O3 concentration. In contrast, the exceedance of the concentration-based critical level of O3 will increase with the projected increase in tropospheric background O3 concentration. The influence of climate change should be considered when predicting the future effects of O3 on vegetation. There is a clear need for multi-factorial, open-air experiments to provide more realistic information for O3 flux-effect modelling in a future climate.     

Ozone flux to Picea sitchensis (Bong) Carr and Picea abies (L) Karst during short episodes and the effects of these on transpiration and photosynthesis

Sitka spruce and Norway spruce were grown in controlled environments and then exposed to ozone (O3) for short periods as in mid-afternoon episodes experienced in the forest. For concentrations of between 20 and 300 nl litre(-1) there were linear relationships between exposure concentration and O3 uptake rates. Increasing photon flux densities increased rates of photosynthesis and transpiration, the increases being larger in actively growing than dormant seedlings. Physiological condition (dormancy or active growth), species and photon flux density were found to influence O3 flux via their effects on stomatal conductance. Exposure to 80 nl litre(-1) O3 resulted in consistent increases of stomatal conductance and there were also indications that water-use efficiency was decreased.     

Methane emissions from wastewater management

Gas exchange and ozone-induced foliar injury were intensively measured during a 6-day period in mid-August 1998 on leaves of Acer pseudoplatanus, Betula pendula, Corylus avellana, Fagus sylvatica, Fraxinus excelsior, Morus nigra, Prunus avium, Prunus serotina, Rhamnus cathartica, and Viburnum lantana at a forest nursery site in Canton Ticino, Switzerland. Plants were grown in four open plots (AA), four open-top chambers receiving carbon-filtered (CF) air, and four receiving non-filtered (NF) air. Significant variation in gas exchange (F > 12.7, P < 0.001) was detected among species with average net photosynthesis and average stomatal conductance differing by a factor of two. Species also varied significantly in foliar injury for those leaves for which we measured gas exchange (F = 39.6, P < 0.001). Fraxinus excelsior, M. nigra, P. avium, P. serotina, R. cathartica, and V. lantana showed more injury than A. pseudoplatanus, B. pendula, C. avellana, and Fagus sylvatica. Plants grown in CF chambers had significantly higher net photosynthesis (A) and stomatal conductance to water vapor (gwv), and lower foliar injury than plants grown in NF chambers and AA plots; interactions between species and ozone treatments were significant for all variables (F > or = 2.2, P < 0.05) except gwv (F = 0.7, P > 0.1). Although A and gwv decreased and foliar injury increased with leaf age, the magnitude of these changes was lower for plants grown in CF chambers than for plants grown in NF chambers and AA plots. Neither ozone uptake threshold (r = 0.26, P > 0.20) nor whole-plant injury (r = -0.15, P > 0.41) was significantly correlated with stomatal conductance across these species. It appears that the relationships between stomatal conductance and foliar injury are species-specific and interactions between physiology and environments and leaf biochemical processes must be considered in determining species sensitivity to ambient ozone exposures.     

Size-mediated foliar response to ozone in black cherry trees

Local ozone concentration and visible foliar injury were measured over the 1994 growing season on open-grown black cherry (Prunus serotina Ehrh.) trees of varying size (age) within forest stands and adjacent openings at a site in north-central Pennsylvania. Relationships were determined between visible ozone injury and ozone exposure, as well as calculated between injury and ozone uptake expressed as the product of stomatal conductance and ozone concentration. In addition, simultaneous measurements of visible symptoms and leaf gas exchange were also conducted to determine the correlation between visible and physiological injury and ozone exposure. By September, the amount of leaf area affected by visible foliar ozone injury was greatest for seedlings (46%), followed by canopy trees (20%) and saplings (15%). A large amount of variability in foliar ozone symptom expression was observed among trees within a size class. Sum40 and Sum60 (ozone concentration > 40 and > 60 nl liter(-1)) cumulative exposure statistics were the most meaningful indices for interpretation of foliar injury response. Seedlings were apparently more sensitive to ozone injury than larger trees because their higher rates of stomatal conductance resulted in higher rates of ozone uptake. Seedlings also had higher rates of early leaf abscission than larger trees with an average of nearly 30% of the leaves on a shoot abscised by 1 September compared to approximately 5% for larger trees. However, per unit ozone uptake into the leaf, larger trees exhibited larger amounts of foliar injury. The amount of visible foliar injury was negatively correlated (r(2) = 0.82) with net photosynthetic rates, but was not related to stomatal conductance. Net photosynthesis and stomatal conductance thus became uncoupled at high levels of visible foliar injury.     

Ozone fluxes above and within a pine forest canopy in dry and wet conditions

The physiological and physical processes controlling ozone dry deposition to vegetated surfaces are still not fully understood. In particular, the role of the understorey and the possible action of dew on ozone deposition have not received much attention so far. This paper presents the results of an experiment aimed at quantifying ozone dry deposition to a maritime pine forest in the “Les Landes” area in France. Ozone deposition fluxes were measured using the eddy-covariance technique above and within the canopy. We investigate the factors acting on ozone deposition in both dew-wetted and dry conditions. The values obtained for the ozone deposition velocity are well in the range of previously published measurements over coniferous forests. For the present forest, ozone uptake by the understorey is a significant portion of ozone deposition to the whole pine stand. The understorey contributes more to the overall ozone flux than to the other measured scalar fluxes (sensible heat and water vapour). During dry nights the surface conductance for ozone and the friction velocity are strongly correlated, showing that ozone deposition is largely controlled by dynamical processes. During the day, in dry conditions, the canopy stomatal conductance is the major parameter controlling ozone deposition. However, in winter, when the stomatal conductance is low, the influence of dynamical processes persists during day-time. It is also found that surface wetness associated with dew significantly enhances ozone deposition, during the night as well as in the morning.     

Seasonal and diurnal gas exchange differences in ozone-sensitive common milkweed (Asclepias syriaca L.) in relation to ozone uptake

Stomatal conductance and net photosynthesis of common milkweed (Asclepias syriaca L.) plants in two different soil moisture regimes were directly quantified and subsequently modeled over an entire growing season. Direct measurements captured the dynamic response of stomatal conductance to changing environmental conditions throughout the day, as well as declining gas exchange and carbon assimilation throughout the growth period beyond an early summer maximum. This phenomenon was observed in plants grown both with and without supplemental soil moisture, the latter of which should theoretically mitigate against harmful physiological effects caused by exposure to ozone. Seasonally declining rates of stomatal conductance were found to be substantial and incorporated into models, making them less susceptible to the overestimations of effective exposure that are an inherent source of error in ozone exposure indices. The species-specific evidence presented here supports the integration of dynamic physiological processes into flux-based modeling approaches for the prediction of ozone injury in vegetation.     

Ozone sensitivity of Fagus sylvatica and Fraxinus excelsior young trees in relation to leaf structure and foliar ozone uptake

During the summer of 2001, 2-year-old Fraxinus excelsior and Fagus sylvatica plants were subjected to ozone-rich environmental conditions at the Regional Forest Nursery at Curno (Northern Italy). Atmospheric ozone concentrations and stomatal conductance were measured, in order to calculate the foliar fluxes by means of a one-dimensional model. The foliar structure of both species was examined (thickness of the lamina and of the individual tissues, leaf mass per area, leaf density) and chlorophyll a fluorescence was determined as a response parameter. Stomatal conductance was always greater in Fraxinus excelsior, as was ozone uptake, although the highest absorption peaks did not match the peaks of ozone concentration in the atmosphere. The foliar structure can help explain this phenomenon: Fraxinus excelsior has a thicker mesophyll than Fagus sylvatica (indicating a greater photosynthesis potential) and a reduced foliar density. This last parameter, related to the apoplastic fraction, suggests a greater ability to disseminate the gases within the leaf as well as a greater potential detoxifying capacity. As foliar symptoms spread, the parameters relating to chlorophyll a fluorescence also change. PI (Performance Index, Strasser, A., Srivastava, A., Tsimilli-Michael, M., 2000. The fluorescence transient as a tool to characterize and screen photosynthetic samples. In: Yunus, M., Pathre, U., Mohanty, P., (Eds.) Probing Photosynthesis: Mechanisms, Regulation and Adaptation. Taylor & Francis, London, UK, pp. 445-483.) has proved to be a more suitable index than Fv/Fm (Quantum Yield Efficiency) to record the onset of stress conditions.     

Modelling stomatal ozone flux and deposition to grassland communities across Europe

Regional scale modelling of both ozone deposition and the risk of ozone impacts is poorly developed for grassland communities. This paper presents new predictions of stomatal ozone flux to grasslands at five different locations in Europe, using a mechanistic model of canopy development for productive grasslands to generate time series of leaf area index and soil water potential as inputs to the stomatal component of the DO(3)SE ozone deposition model. The parameterisation of both models was based on Lolium perenne, a dominant species of productive pasture in Europe. The modelled seasonal time course of stomatal ozone flux to both the whole canopy and to upper leaves showed large differences between climatic zones, which depended on the timing of the start of the growing season, the effect of soil water potential, and the frequency of hay cuts. Values of modelled accumulated flux indices and the AOT40 index showed a five-fold difference between locations, but the locations with the highest flux differed depending on the index used; the period contributing to the accumulation of AOT40 did not always coincide with the modelled period of active ozone canopy uptake. Use of a fixed seasonal profile of leaf area index in the flux model produced very different estimates of annual accumulated total canopy and leaf ozone flux when compared with the flux model linked to a simulation of canopy growth. Regional scale model estimates of both the risks of ozone impacts and of total ozone deposition will be inaccurate unless the effects of climate and management in modifying grass canopy growth are incorporated.     

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.     

Ozone induced leaf loss and decreased leaf production of European Holly (Ilex aquifolium L.) over multiple seasons

European Holly (Ilex aquifolium L.) was used to study the impact of one short (28 day) ozone fumigation episode on leaf production, leaf loss and stomatal conductance (g(s)), in order to explore potential longer term effects over 3 growing seasons. Young I. aquifolium plants received an episode of either charcoal-filtered air or charcoal-filtered air with 70 nl l(-1) O(3) added for 7 h d(-1) over a 28 day period from June 15th 1996, then placed into ambient environment, Stoke-on-Trent, U.K. Data were collected per leaf cohort over the next three growing seasons. Ozone exposure significantly increased leaf loss and stomatal conductance and reduced leaf production over all subsequent seasons. Impact of the initial ozone stress was still detected in leaves that had no direct experimental ozone exposure. This study has shown the potential of ozone to introduce long-term phenological perturbations into ecosystems by influencing productivity over a number of seasons.