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.     

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.     

Physiology, morphology, and ozone uptake of leaves of black cherry seedlings, saplings, and canopy trees

Patterns of ozone uptake were related to physiological, morphological, and phenological characteristics of different-sized black cherry trees (Prunus serotina Ehrh.) at a site in central Pennsylvania. Calculated ozone uptake differed among open-grown seedlings, forest gap saplings, and canopy trees and between leaves in the upper and lower crown of saplings and canopy trees. On an instantaneous basis, seedling leaves had the greatest ozone uptake rates of all tree size classes due to greater stomatal conductance and higher concentrations of ozone in their local environment. A pattern of higher stomatal conductance of seedlings was consistent with higher incident photosynthetically-active radiation, stomatal density, and predawn xylem water potentials for seedlings relative to larger trees. However, seedlings displayed an indeterminate pattern of shoot growth, with the majority of their leaves produced after shoot growth had ceased for canopy and sapling trees. Full leaf expansion occurred by mid-June for sapling and canopy trees. Because many of their leaves were exposed to ozone for only part of the growing season, seedlings had a lower relative exposure over the course of the growing season, and subsequently lower cumulative uptake, of ozone than canopy trees and a level of uptake similar to upper canopy leaves of saplings. Visible injury symptoms were not always correlated with patterns in ozone uptake. Visible symptoms were more apparent on seedling leaves in concurrence with their high instantaneous uptake rates. However, visible injury was more prevalent on leaves in the lower versus upper crown of canopy trees and saplings, even though lower crown leaves had less ozone uptake. Lower crown leaves may be more sensitive to ozone per unit uptake than upper crown leaves because of their morphology. In addition, the lower net carbon uptake of lower crown leaves may limit repair and anti-oxidant defense processes.     

Quantification of ozone uptake at the stand level in a Pinus canariensis forest in Tenerife, Canary Islands: an approach based on sap flow measurements

Ozone uptake was studied in a pine forest in Tenerife, Canary Islands, an ecotone with strong seasonal changes in climate. Ambient ozone concentration showed a pronounced seasonal course with high concentrations during the dry and warm period and low concentrations during the wet and cold season. Ozone uptake by contrast showed no clear seasonal trend. This is because canopy conductance significantly decreased with soil water availability and vapour pressure deficit. Mean daily ozone uptake averaged 1.9 nmol m(-2) s(-1) during the wet and cold season, and 1.5 nmol m(-2) s(-1) during the warm and dry period. The corresponding daily mean ambient ozone concentrations were 42 and 51 nl l(-1), respectively. Thus we conclude that in Mediterranean type forest ecosystems the flux based approach is more capable for risk assessment than an external, concentration based approach.     

Seasonal variation of gas exchange and pigmentation in branches of three grafted clones of mature ponderosa pine exposed to ozone and acid rain

Gas exchange and pigmentation responses of mature ponderosa pine (Pinus ponderosa Laws.) branches to ozone and acid rain exposure were investigated using three grafted clones growing in a managed seed orchard. Exposure of one-year-old foliage to twice ambient ozone (2 x AMB) resulted in significant decreases in net photosynthesis (Pn), stomatal conductance (gsw) and pigmentation relative to charcoal-filtered (CF) and ambient (AMB) ozone treatments. Ozone effects on gas exchange and pigmentation were most pronounced during late-season and differed significantly among clones. Environmental parameters (e.g. light, vapor pressure deficit, and temperature) accounted for more variation in Pn than did cumulative ozone exposure. Minimal differences in gsw and Pn among ozone treatments occurred during seasonal periods of high temperature and evaporative demand. Negative effects of 2 x AMB ozone on gsw and pigmentation were greatest for the clones having highest and lowest phenotypic vigor under ambient conditions; the clone of moderate phenotypic vigor under ambient conditions was least sensitive to ozone. Application of simulated acid rain of pH 3.0, pH 5.1 or no rain (NR) had little impact on gas exchange or pigmentation.     

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.     

Simulations of stomatal conductance and ozone uptake to Norway spruce saplings in open-top chambers

A simulation model was developed to estimate the stomatal conductance and ozone flux to Norway spruce saplings in open-top chambers. The model was parameterized against needle conductance measurements that were made on 4-6-year-old spruce saplings, grown in open-top chambers, in July-September during three different seasons. The spruce saplings were either maintained well watered or subject to a 7-8 week drought period in July-September each year. The simulated conductance showed a good agreement with the measured conductance for the well-watered as well as the drought stress-treated saplings. The simulations were significantly improved when different vapour pressure deficit (VPD) functions were applied for well-watered and drought-stressed spruce saplings. The cumulated ozone uptake which was calculated from the conductance simulations showed less variation between years, compared to the cumulative ozone exposure index AOT40 (accumulated exposure over a threshold of 40 ppb or nl l(-1)) for the corresponding time periods. Measurements in May 1995 demonstrated the occurrence of long-term 'memory-effects' from the drought stress treatments on the conductance. Memory-effects need to be considered when simulation models for stomatal conductance are to be applied to long-lived forest trees under a multiple stress situation.     

Physiological and foliar symptom response in the crowns of Prunus serotina, Fraxinus americana and Acer rubrum canopy trees to ambient ozone under forest conditions

The crowns of five canopy dominant black cherry (Prunus serotina Ehrh.), five white ash (Fraxinus americana L.), and six red maple (Acer rubrum L.) trees on naturally differing environmental conditions were accessed with scaffold towers within a mixed hardwood forest stand in central Pennsylvania. Ambient ozone concentrations, meteorological parameters, leaf gas exchange and leaf water potential were measured at the sites during the growing seasons of 1998 and 1999. Visible ozone-induced foliar injury was assessed on leaves within the upper and lower crown branches of each tree. Ambient ozone exposures were sufficient to induce typical symptoms on cherry (0-5% total affected leaf area, LAA), whereas foliar injury was not observed on ash or maple. There was a positive correlation between increasing cumulative ozone uptake (U) and increasing percent of LAA for cherry grown under drier site conditions. The lower crown leaves of cherry showed more severe foliar injury than the upper crown leaves. No significant differences in predawn leaf water potential (psi(L)) were detected for all three species indicating no differing soil moisture conditions across the sites. Significant variation in stomatal conductance for water vapor (g(wv)) was found among species, soil moisture, time of day and sample date. When comparing cumulative ozone uptake and decreased photosynthetic activity (P(n)), red maple was the only species to show higher gas exchange under mesic vs. drier soil conditions (P < 0.05). The inconsistent differences in gas exchange response within the same crowns of ash and the uncoupling relationship between g(wv) and P(n) demonstrate the strong influence of heterogeneous environmental conditions within forest canopies.     

Increasing risk for negative ozone impacts on vegetation in northern Sweden

Trends were found for increasing surface ozone concentrations during April-September in northern Sweden over the period 1990-2006 as well as for an earlier onset of vegetation growing season. The highest ozone concentrations in northern Sweden occurred in April and the ozone concentrations in April showed a strong increasing trend. A model simulation of ozone flux for Norway spruce indicated that the provisional ozone flux based critical level for forests in Europe is exceeded in northern Sweden. Future climate change would have counteracting effects on the stomatal conductance and needle ozone uptake, mediated on the one hand by direct effect of increasing air temperatures and on the other through increasing water vapour pressure difference between the needles and air. Thus, there is a substantial and increasing risk for negative impacts of ozone on vegetation in northern Sweden, related mainly to increasing ozone concentrations and an earlier onset of the growing season.     

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.     

Assessing the risk caused by ground level ozone to European forest trees: a case study in pine, beech and oak across different climate regions

Two different indices have been proposed for estimation of the risk caused to forest trees across Europe by ground-level ozone, (i) the concentration based AOT40 index (Accumulated Over a Threshold of 40 ppb) and (ii) the recently developed flux based AFstY index (Accumulated stomatal Flux above a flux threshold Y). This paper compares the AOT40 and AFstY indices for three forest trees species at different locations in Europe. The AFstY index is estimated using the DO(3)SE (Deposition of Ozone and Stomatal Exchange) model parameterized for Scots pine (Pinus sylvestris), beech (Fagus sylvatica) and holm oak (Quercus ilex). The results show a large difference in the perceived O(3) risk when using AOT40 and AFstY indices both between species and regions. The AOT40 index shows a strong north-south gradient across Europe, whereas there is little difference between regions in the modelled values of AFstY. There are significant differences in modelled AFstY between species, which are predominantly determined by differences in the timing and length of the growing season, the periods during which soil moisture deficit limits stomatal conductance, and adaptation to soil moisture stress. This emphasizes the importance of defining species-specific flux response variables to obtain a more accurate quantification of O(3) risk.     

Growth of soybean at future tropospheric ozone concentrations decreases canopy evapotranspiration and soil water depletion

Tropospheric ozone is increasing in many agricultural regions resulting in decreased stomatal conductance and overall biomass of sensitive crop species. These physiological effects of ozone forecast changes in evapotranspiration and thus in the terrestrial hydrological cycle, particularly in intercontinental interiors. Soybean plots were fumigated with ozone to achieve concentrations above ambient levels over five growing seasons in open-air field conditions. Mean season increases in ozone concentrations ([O₃]) varied between growing seasons from 22 to 37% above background concentrations. The objective of this experiment was to examine the effects of future [O₃] on crop ecosystem energy fluxes and water use. Elevated [O₃] caused decreases in canopy evapotranspiration resulting in decreased water use by as much as 15% in high ozone years and decreased soil water removal. In addition, ozone treatment resulted in increased sensible heat flux in all years indicative of day-time increase in canopy temperature of up to 0.7 °C.     

Statistical estimation of ozone exposure metrics

Data from recent experiments at North Carolina State University and other locations provide a unique opportunity to study the effect of ambient ozone on the growth of clover. The data consist of hourly ozone measurements over a 140 day growing season at eight sites in the US, coupled with clover growth response data measured every 28 days. The objective is to model an indicator of clover growth as a function of ozone exposure. A common strategy for dealing with the numerous hourly ozone measurements is to reduce these to a single summary measurement, a so-called exposure metric, for the growth period of interest. However, the mean ozone value is not necessarily the best summarization, as it is widely believed that low levels of ozone have a negligible effect on growth, whereas peak ozone values are deleterious to plant growth. There are also suspected interactions with available sunlight, temperature and humidity. A number of exposure metrics have been proposed that reflect these beliefs by assigning different weights to ozone values according to magnitude, time of day, temperature and humidity. These weighting schemes generally depend on parameters that have, to date, been subjectively determined. We propose a statistical approach based on profile likelihoods to estimate the parameters in these exposure metrics.     

Impact of increased springtime O3 exposure on Scots pine (Pinus sylvestris) seedlings in central Finland

Three-year-old Scots pine (Pinus sylvestris L.) seedlings were exposed to ambient or elevated ozone (O(3)) concentrations in open-air exposure fields in central Finland in 1995-97. Three different treatments were applied in 1996 and 1997: ambient air, elevated O(3) (1.3-1.5xambient) during the growing season (June-September) and elevated O(3) in March-September, i.e. the growing season including the springtime O(3) exposure. The ambient mean O(3) concentrations were 40% higher in springtime (March-May) compared to the concentrations during the growing seasons. Maximum O(3) concentrations were measured in April or early May, whereas a clear increase in the stomatal activity of the seedlings was observed by the middle of May. This suggests a low intake of O(3) by conifers despite the higher O(3) concentrations in spring. Stomatal conductance, and contents of chlorophyll and ribulosebisphosphate carboxylase/oxygenase (Rubisco) in current-year needles were not significantly affected by any O(3) treatment. Only a slight decrease in current-year shoot growth, slight increase in the abscission of 2-year-old needles and increased electron density of chloroplast stroma by springtime O(3) exposure suggest a rather small contribution of elevated springtime O(3) concentrations to total O(3) damage under current climatic conditions in Finland. However, the increases in springtime O(3) concentrations may enhance the cumulative effects of O(3) during long-term O(3) exposures.     

Modelling stomatal ozone flux across Europe

A model has been developed to estimate stomatal ozone flux across Europe for a number of important species. An initial application of this model is illustrated for two species, wheat and beech. The model calculates ozone flux using European Monitoring and Evaluation Programme (EMEP) model ozone concentrations in combination with estimates of the atmospheric, boundary layer and stomatal resistances to ozone transfer. The model simulates the effect of phenology, irradiance, temperature, vapour pressure deficit and soil moisture deficit on stomatal conductance. These species-specific microclimatic parameters are derived from meteorological data provided by the Norwegian Meteorological Institute (DNMI), together with detailed land-use and soil type maps assembled at the Stockholm Environment Institute (SEI). Modelled fluxes are presented as mean monthly flux maps and compared with maps describing equivalent values of AOT40 (accumulated exposure over threshold of 40 ppb or nl l(-1)), highlighting the spatial differences between these two indices. In many cases high ozone fluxes were modelled in association with only moderate AOT40 values. The factors most important in limiting ozone uptake under the model assumptions were vapour pressure deficit (VPD), soil moisture deficit (for Mediterranean regions in particular) and phenology. The limiting effect of VPD on ozone uptake was especially apparent, since high VPDs resulting in stomatal closure tended to co-occur with high ozone concentrations. Although further work is needed to link the ozone uptake and deposition model components, and to validate the model with field measurements, the present results give a clear indication of the possible implications of adopting a flux-based approach for future policy evaluation.     

Responses of hybrid poplar clones and red maple seedlings to ambient O(3) under differing light within a mixed hardwood forest

The responses of ramets of hybrid poplar (Populus spp.) (HP) clones NE388 and NE359, and seedlings of red maple (Acer rubrum, L.) to ambient ozone (O(3)) were studied during May-September of 2000 and 2001 under natural forest conditions and differing natural sunlight exposures (sun, partial shade and full shade). Ambient O(3) concentrations at the study site reached hourly peaks of 109 and 98 ppb in 2000 and 2001, respectively. Monthly 12-h average O(3) concentrations ranged from 32.3 to 52.9 ppb. Weekly 12-h average photosynthetically active radiation (PAR) within the sun, partial shade and full shade plots ranged from 200 to 750, 50 to 180, and 25 to 75 micromol m(-2) s(-1), respectively. Ambient O(3) exposure induced visible foliar symptoms on HP NE388 and NE359 in both growing seasons, with more severe injury observed on NE388 than on NE359. Slight foliar symptoms were observed on red maple seedlings during the 2001 growing season. Percentage of total leaf area affected (%LAA) was positively correlated with cumulative O(3) exposures. More severe foliar injury was observed on plants grown within the full shade and partial shade plots than those observed on plants grown within the sun plot. Lower light availability within the partial shade and full shade plots significantly decreased net photosynthetic rate (Pn) and stomatal conductance (g(wv)). The reductions in Pn were greater than reductions in g(wv), which resulted in greater O(3) uptake per unit Pn in plants grown within the partial shade and full shade plots. Greater O(3) uptake per unit Pn was consistently associated with more severe visible foliar injury in all species and/or clones regardless of differences in shade tolerance. These studies suggest that plant physiological responses to O(3) exposure are likely complicated due to multiple factors under natural forest conditions.     

Modelling of stomatal conductance and ozone flux of Norway spruce: comparison with field data

It has been proposed that stomatal flux of ozone would provide a more reliable basis than ozone exposure indices for the assessment of the risk of ozone damage to vegetation across Europe. However, implementation of this approach requires the development of appropriate models which need to be rigorously tested against actual data collected under field conditions. This paper describes such an assessment of the stomatal component of the model described by Emberson et al. (2000. Modelling stomatal ozone flux across Europe. Environmental Pollution 110). Model predictions are compared with field measurements of both stomatal conductance (g(s)) and calculated ozone flux for shoots of mature Norway spruce (Picea abies) growing in the Tyrol Mountains in Austria. The model has been developed to calculate g(s) as a function of leaf phenology and four environmental variables: photosynthetic flux density (PFD), temperature, vapour pressure deficit (VPD) and soil moisture deficit (SMD). The model was run using climate data measured on site, although the SMD component was omitted since the necessary data were not available. The model parameterisation for Norway spruce had previously been collected from the scientific literature and therefore established independently from the measurement study. Overall, strong associations were found between model predictions and measured values of stomatal conductance to ozone (GO(3)) and calculated stomatal ozone flux (FO(3)). Average diurnal profiles of GO(3) and FO(3) showed good agreement between the field data and modelled values except during the morning period of 1990. The diurnal pattern of ozone flux was determined primarily by PFD and VPD, as there was little diurnal variation in ozone concentration. In general, the model predicted instances of high ozone flux satisfactorily, indicating its potential applicability in identifying areas of high ozone risk for this species.     

A cumulative ozone uptake-response relationship for the growth of Norway spruce saplings

Norway spruce saplings [Picea abies (L.) Karst.] were exposed during four growing seasons to different ozone treatments in open-top chambers: charcoal filtered air (CF), non-filtered air (NF) and non-filtered air with extra ozone (NF+, 1.4xambient concentrations). The CF and NF+ ozone treatments were combined with phosphorous deficiency and drought stress treatments. The total biomass of the trees was harvested at different intervals during the experimental period. The ozone uptake to current-year needles of the Norway spruce saplings was estimated using a multiplicative stomatal conductance simulation model. There was a highly significant correlation between the reduction of total biomass and the estimated cumulative ozone uptake, which did not vary when different thresholds were applied for the rate of ozone uptake. The reduction of the total biomass was estimated to 1% per 10 mmol m(-2) cumulated ozone uptake, on a projected needle area basis.     

Relations between crown condition and ozone and its dependence on environmental factors

It is now widely accepted that stomatal ozone uptake provides a more reliable indicator for the assessment of risk of ozone damage to forest trees than ozone exposure. Thus, environmental factors influencing stomatal opening strongly affect the sensitivity of forest trees towards high ozone concentrations. In this study the hydrological model WAWAHAMO is used to simulate stomatal opening dependent on air temperature, solar radiation, vapor pressure deficit and soil water potential. Then, a statistical analysis was carried out to investigate relations between crown defoliation and ozone concentrations dependent on stomatal opening. The results support the hypothesis that tropospheric ozone contributes to the observed crown defoliation. However, environmental factors strongly affect tree response to high ozone concentrations.     

Use of sap flow measurements to validate stomatal functions for mature beech (Fagus sylvatica) in view of ozone uptake calculations

For a quantitative estimate of the ozone effect on vegetation reliable models for ozone uptake through the stomata are needed. Because of the analogy of ozone uptake and transpiration it is possible to utilize measurements of water loss such as sap flow for quantification of ozone uptake. This technique was applied in three beech (Fagus sylvatica) stands in Switzerland. A canopy conductance was calculated from sap flow velocity and normalized to values between 0 and 1. It represents mainly stomatal conductance as the boundary layer resistance in forests is usually small. Based on this relative conductance, stomatal functions to describe the dependence on light, temperature, vapour pressure deficit and soil moisture were derived using multivariate nonlinear regression. These functions were validated by comparison with conductance values directly estimated from sap flow. The results corroborate the current flux parameterization for beech used in the DO₃SE model.