Leaf morphology and ozone sensitivity of two open pollinated genotypes of black cherry (Prunus serotina) seedlings

Two black cherry families differing in sensitivity to ozone (O(3)) were used to test the hypothesis that leaf morphology is related to foliar sensitivity to O(3). Two uninjured sections of leaf tissue were sampled from a single leaf collected from each of 12 open-grown O(3)-tolerant and 12 O(3)-sensitive seedlings. Standard histological techniques and light microscopy were used to examine 11 morphological characteristics. The O(3)-sensitive genotype had significantly greater stomatal density, thinner palisade mesophyll layer and thicker spongy mesophyll layer, lower ratio of palisade to spongy mesophyll, greater leaf weight and leaf area. In addition, total leaf thickness, guard cell length, and specific leaf mass were also slightly greater for the O(3)-sensitive genotype.     

Physiological and foliar injury responses of Prunus serotina,Fraxinus americana,and Acer rubrum seedlings to varying soil moisture and ozone

Sixteen black cherry (Prunus serotina, Ehrh.), 10 white ash (Fraxinus americana, L.) and 10 red maple (Acer rubrum, L.) 1-year old seedlings were planted per plot in 1997 on a former nursery bed within 12 open-top chambers and six open plots. Seedlings were exposed to three different ozone scenarios (ambient air: 100% O3; non-filtered air: 98% ambient O3; charcoal-filtered air: 50% ambient O3) within each of two different water regimes (nine plots irrigated, nine plots non-irrigated) during three growing seasons.During the 1998 and 1999 growing season, leaf gas exchange, plant water relations, and foliar injury were measured. Climatic data,ambient- and chamber-ozone-concentrations were monitored. We found that seedlings grown under irrigated conditions had similar (in 1998) but significantly higher gas exchange rates (in 1999) than seedlings grown within non-irrigated plots among similar ozone exposures. Cherry and ash had similar ozone uptake but cherry developed more ozone-induced injury (< 34% affected leaf area, LAA) than ash (<5% LAA), while maple rarely showed foliar injury, indicating the species differed in ozone sensitivity. Significantly more severe injury on seedlings grown under irrigated conditions than seedlings grown under non-irrigated conditions demonstrated that soil moisture altered seedling responses to ambient ozone exposures.     

Field responses of Prunus serotina and Asclepias syriaca to ozone around southern Lake Michigan

Higher ozone concentrations east of southern Lake Michigan compared to west of the lake were used to test hypotheses about injury and growth effects on two plant species. We measured approximately 1000 black cherry trees and over 3000 milkweed stems from 1999 to 2001 for this purpose. Black cherry branch elongation and milkweed growth and pod formation were significantly higher west of Lake Michigan while ozone injury was greater east of Lake Michigan. Using classification and regression tree (CART) analyses we determined that departures from normal precipitation, soil nitrogen and ozone exposure/peak hourly concentrations were the most important variables affecting cherry branch elongation, and milkweed stem height and pod formation. The effects of ozone were not consistently comparable with the effects of soil nutrients, weather, insect or disease injury, and depended on species. Ozone SUM06 exposures greater than 13 ppm-h decreased cherry branch elongation 18%; peak 1-h exposures greater than 93 ppb reduced milkweed stem height 13%; and peak 1-h concentrations greater than 98 ppb reduced pod formation 11% in milkweed.     

Observations of ozone-induced foliar injury on black cherry (Prunus serotina, var. capuli) within the Desierto de Los Leones National Park, Mexico City

A survey for ozone-induced foliar injury of black cherry was conducted in mid-June 1995 within the Desierto de Los Leones National Park located southwest of Mexico City. Evaluations of the upper and lower tree crowns of 18 trees revealed evidence of significant upper surface stipple, leaf reddening and premature senescence on 72% of the trees. A general survey of an additional 169 trees disclosed that 41% exhibited similar symptoms. A gradient of increasing symptoms with increasing elevation was also evident. For the most part, asymptomatic trees were observed to be situated within well-shaded coves at the lower elevations with very few symptomatic trees present in these areas.     

Seasonal trends in reduced leaf gas exchange and ozone-induced foliar injury in three ozone sensitive woody plant species

Seasonal trends in leaf gas exchange and ozone-induced visible foliar injury were investigated for three ozone sensitive woody plant species. Seedlings of Populus nigra L., Viburnum lantana L., and Fraxinus excelsior L. were grown in charcoal-filtered chambers, non-filtered chambers and open plots. Injury assessments and leaf gas exchange measurements were conducted from June to October during 2002. All species developed typical ozone-induced foliar injury. For plants exposed to non-filtered air as compared to the charcoal-filtered air, mean net photosynthesis was reduced by 25%, 21%, and 18% and mean stomatal conductance was reduced by 25%, 16%, and 8% for P. nigra, V. lantana, and F. excelsior, respectively. The timing and severity of the reductions in leaf gas exchange were species specific and corresponded to the onset of visible foliar injury.     

Effects of ozone on growth, yield and leaf gas exchange rates of four Bangladeshi cultivars of rice (Oryza sativa L.)

To assess the effects of tropospheric O₃ on rice cultivated in Bangladesh, four Bangladeshi cultivars (BR11, BR14, BR28 and BR29) of rice (Oryza sativa L.) were exposed daily to charcoal-filtered air or O₃ at 60 and 100 nl l⁻¹ (10:00-17:00) from 1 July to 28 November 2008. The whole-plant dry mass and grain yield per plant of the four cultivars were significantly reduced by the exposure to O₃. The exposure to O₃ significantly reduced net photosynthetic rate of the 12th and flag leaves of the four cultivars. The sensitivity to O₃ of growth, yield and leaf gas exchange rates was not significantly different among the four cultivars. The present study suggests that the sensitivity to O₃ of yield of the four Bangladeshi rice cultivars is greater than that of American rice cultivars and is similar to that of Japanese rice cultivars and that O₃ may detrimentally affect rice production in Bangladesh.     

Effects of ozone on growth, yield and leaf gas exchange rates of two Bangladeshi cultivars of wheat (Triticum aestivum L.)

To clarify the effects of O₃ on crop plants cultivated in Bangladesh, two Bangladeshi wheat cultivars (Sufi and Bijoy) were grown in plastic boxes filled with Andisol and exposed daily to charcoal-filtered air or O₃ at 60 and 100 nl l⁻¹ (10:00-17:00) from 13 March to 4 June 2008. The whole-plant dry mass and grain yield per plant of the two cultivars at the final harvest were significantly reduced by the exposure to O₃. Although there was no significant effect of O₃ on stomatal diffusive conductance to H₂O of flag leaf, net photosynthetic rate of the leaf was significantly reduced by the exposure to O_3. The sensitivity of growth, yield, yield components and leaf gas exchange rates to O₃ was not significantly different between the two cultivars. The results obtained in the present study suggest that ambient levels of O₃ may detrimentally affect wheat production in Bangladesh.     

Leaf age affects the responses of foliar injury and gas exchange to tropospheric ozone in Prunus serotina seedlings

We investigated the effect of leaf age on the response of net photosynthesis (A), stomatal conductance (g_wv), foliar injury, and leaf nitrogen concentration (N_L) to tropospheric ozone (O₃) on Prunus serotina seedlings grown in open-plots (AA) and open-top chambers, supplied with either carbon-filtered or non-filtered air. We found significant variation in A, g_wv, foliar injury, and N_L (P < 0.05) among O₃ treatments. Seedlings in AA showed the highest A and g_wv due to relatively low vapor pressure deficit (VPD). Older leaves showed significantly lower A, g_wv, N_L, and higher foliar injury (P < 0.001) than younger leaves. Leaf age affected the response of A, g_wv, and foliar injury to O₃. Both VPD and N_L had a strong influence on leaf gas exchange. Foliar O₃-induced injury appeared when cumulative O₃ uptake reached 8-12 mmol m⁻², depending on soil water availability. The mechanistic assessment of O₃-induced injury is a valuable approach for a biologically relevant O₃ risk assessment for forest trees.     

Visible injury, crown condition, and growth responses of selected Italian forests in relation to ozone exposure

The impact of ozone on forest ecosystems in Italy is monitored within the CONECOFOR programme. Ozone levels are measured in 30 plots using passive samplers. Response parameters used are: crown condition (transparency), BAI (basal area increment), and visible symptoms on spontaneous vegetation. Levels of AOT40 are above the concentration-based critical level of 5 ppmh in all sites, but the evidence of impact on forest vegetation remains limited. Ozone is a predictor of crown transparency residuals in beech sites over two consecutive years, but the variance explained amounts to less than 10%. The relation between BAI reduction and ozone is even less certain. Transparency and BAI are more readily explainable in terms of ecological conditions of the site and climate fluctuations. The interpretation of visible symptoms is doubtful, and is conditioned by the prevailing ecological factors in the areas.     

Effects of chronic elevated ozone exposure on gas exchange responses of adult beech trees (Fagus sylvatica) as related to the within-canopy light gradient

The effects of elevated O₃ on photosynthetic properties in adult beech trees (Fagus sylvatica) were investigated in relation to leaf mass per area as a measure of the gradually changing, within-canopy light availability. Leaves under elevated O₃ showed decreased stomatal conductance at unchanged carboxylation capacity of Rubisco, which was consistent with enhanced δ¹³C of leaf organic matter, regardless of the light environment during growth. In parallel, increased energy demand for O₃ detoxification and repair was suggested under elevated O₃ owing to enhanced dark respiration. Only in shade-grown leaves, light-limited photosynthesis was reduced under elevated O₃, this effect being accompanied by lowered F_v/F_m. These results suggest that chronic O₃ exposure primarily caused stomatal closure to adult beech trees in the field regardless of the within-canopy light gradient. However, light limitation apparently raised the O₃ sensitivity of photosynthesis and accelerated senescence in shade leaves.     

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.     

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.     

Reduction of stem growth and site dependency of leaf injury in Massachusetts black cherries exhibiting ozone symptoms

Leaf ozone symptoms in natural ecosystems are increasingly reported but ozone effects on tree growth and the mediation of site conditions are still little documented. This study tests two hypotheses: (1) leaf injury in black cherry is associated with decline in radial growth, (2) symptoms are more prevalent on mesic sites. On sites supporting black cherry across Massachusetts, tree growth and leaf ozone injury were surveyed in 1996 using a randomized plot network established in the 1960s. Forty-seven percent of 120 trees sampled for ozone symptoms were symptomatic with generally low levels of injury. Over a 31-year period symptomatic trees had 28% lower stem growth rates than asymptomatic trees. Ozone symptom expression was enhanced in well growing stands on moister, cooler and more elevated sites. Ozone appeared to increase environmental stress and had a more pronounced effect on growth in better growing black cherry stands. This complicates management decisions as thinning increases growth and moisture availability.     

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.     

Differences in growth, leaf senescence and injury, and stomatal density in birch (Betula pendula Roth.) in relation to ambient levels of ozone in Finland

The differences in growth, leaf senescence, visible ozone injuries and stomatal density between one coastal site (natural ozone) and two inland sites (natural and elevated ozone) in Finland were determined for saplings of Betula pendula clones grown under open-field conditions during two growing seasons. Responses in growth, leaf senescence, visible injuries, and stomatal density were determined in relation to cumulative ozone exposure accumulated over the thresholds of 30, 40 and 50 ppb (10(9)) during the exposure period. In addition, the effects of the different ozone exposures on ultrastructure of chloroplasts were studied. Increasing ozone exposure resulted in reduced shoot dry weight, stimulated (first year) or reduced (second year) height growth, accelerated autumn yellowing of leaves, increased stomatal density, visible symptoms and chloroplast injuries, and increased number and size of plastoglobuli. Newly expanded mature leaves in midsummer were more sensitive to ozone episodes than younger developing leaves in the early growing season. In most parameters, the best correlation was achieved with the exposure index AOT30. Ozone risk for birch is highest in the southern coastal area of Finland, where background ozone concentrations are higher than in inland sites.     

Effects of ozone impact on the gas exchange and chlorophyll fluorescence of juvenile birch stems (Betula pendula Roth.)

Effects of ozone impact on gas exchange and chlorophyll fluorescence of juvenile birch (Betula pendula) stems and leaves were investigated. Significant differences in the response of leaves and stems to ozone were found. In leaves, O3 exposure led to a significant decline in photosynthetic rates, whereas stems revealed an increased dark respiration and a concomitant increase in corticular photosynthesis. In contrast to birch leaves, corticular photosynthesis appeared to support the carbon balance of stems or even of the whole-tree under O3 stress. The differences in the ozone-response between leaves and stems were found to be related to ozone uptake rates, and thus to inherent differences in leaf and stem O3 conductance.     

Compensation processes of Aleppo pine (Pinus halepensis Mill.) to ozone exposure and drought stress

A long-term experiment was performed to study the effects of O3 and drought-stress (DS) on Aleppo pine seedlings (Pinus halepensis Mill.) exposed in open-top chambers. Ozone reduced gas exchange rates, ribulose-1,5-biphosphate carboxylase/oxygenase activity (Rubisco), aboveground C and needle N concentrations and C/N ratio and Ca concentrations of the twigs under 3 mm (twigs<3) and the aerial biomass. Also it increased phosphoenolpyruvate carboxylase (PEPc) and N and K concentrations of the twigs<3. Water stress decreased gas exchange rates, predawn needle water potential (PsiPd), C/N ratio, twigs<3 Ca, plant growth, aerial biomass and increased N, twigs with a diameter above 3 mm P and Mg concentrations. The combined exposure to both stresses increased N concentrations of twigs<3 and roots and aboveground biomass K content and decreased root C, maximum daily assimilation rate and instantaneous water use efficiency. The sensitivity of Aleppo pine to both stresses is determined by plant internal resource allocation and compensation mechanisms to cope with stress.     

Ozone uptake (flux) as it relates to ozone-induced foliar symptoms of Prunus serotina and Populus maximowiziixtrichocarpa

Field studies were conducted during 2003 and 2004 from early June to the end of August, at 20 sites of lower or higher elevation within north-central Pennsylvania, using seedlings of black cherry (Prunus serotina, Ehrh.) and ramets of hybrid poplar (Populus maximowiziixtrichocarpa). A linear model was developed to estimate the influence of local environmental conditions on stomatal conductance. The most significant factors explaining stomatal variance were tree species, air temperature, leaf vapor pressure deficit, elevation, and time of day. Overall, environmental factors explained less than 35% of the variation in stomatal conductance. Ozone did not affect gas exchange rates in either poplar or cherry. Ozone-induced foliar injury was positively correlated with cumulative ozone exposures, expressed as SUM40. Overall, the amount of foliar injury was better correlated to a flux-based approach rather than to an exposure-based approach. More severe foliar injuries were observed on plants growing at higher elevations.     

Additive and antagonistic effects of ozone and salinity on the growth, ion contents and gas exchange of five varieties of rice (Oryza sativa L.)

Five varieties of rice (Oryza sativa L.) of varying salinity resistance were grown in non-saline and in saline conditions, with and without a repeated exposure to ozone at a concentration of 83 nmol mol(-1) giving an AOT40 (cumulative exposure above 40 nmol mol(-1)) of 3600 nmol mol(-1) h. Salinity caused a substantial reduction in shoot and root dry weight in all varieties, but the effect on root growth was proportionately less than on shoot growth. Ozone reduced root dry weight but the treatment used did not significantly affect shoot dry weight. Both salinity and ozone reduced plant height. The potassium concentration in the leaves of all five varieties was reduced by salinity, and by ozone in both saline and non-saline treatments. Ozone reduced the sodium concentration in plants grown at 50 mM NaCl but had no effect upon the chloride concentration. Carbon dioxide assimilation, transpiration and stomatal conductance were all reduced by salinity and by ozone and there was close quantitative similarity between the effects of ozone and/or salinity upon assimilation, stomatal conductance and transpiration. There were some antagonistic effects but there were additive effects of salinity and of ozone on root dry weight, plant height, shoot potassium concentration, photosynthesis, transpiration and stomatal conductance. The possible basis of the additive effects of salinity and ozone on gas exchange and mineral uptake are discussed.     

Interactive effects of ozone and elevated carbon dioxide on the growth and physiology of black cherry, green ash, and yellow-poplar seedlings

Potted seedlings of black cherry (Prunus serotina Ehrh.) (BC), green ash (Fraxinus pennsylvanica Marsh.) (GA), and yellow-poplar (Liriodendron tulipifera L.) (YP) were exposed to one of the four treatments: (1) charcoal-filtered air (CF) at ambient CO(2) (control); (2) twice ambient O(3) (2 x O(3)); (3) twice ambient CO(2) (650 microl l(-1)) plus CF air (2 x CO(2)); or (4) twice ambient CO(2) (650 microl l(-1)) plus twice ambient O(3) (2 x CO(2) + 2 x O(3)). The treatments were duplicated in eight continuously stirred tank reactors for 10 weeks. Gas exchange was measured during the last 3 weeks of treatment and all seedlings were destructively harvested after 10 weeks. Significant interactive effects of O(3) and CO(2) on the gas exchange of all three species were limited. The effects of elevated CO(2) and O(3), singly and combined, on light-saturated net photosynthesis (A(max)) and stomatal conductance (g(s)) were inconsistent across species. In all three species, elevated O(3) had no effect on g(s). Elevated CO(2) significantly increased A(max) in GA and YP foliage, and decreased g(s) in YP foliage. Maximum carbon exchange rates and quantum efficiencies derived from light-response curves increased, while compensation irradiance and dark respiration decreased in all three species when exposed to 2 x CO(2). Elevated O(3) affected few of these parameters but any change that was observed was opposite to that from exposure to 2 x CO(2)-air. Interactive effects of CO(2) and O(3) on light-response parameters were limited. Carboxylation efficiencies, derived from CO(2)-response curves (A/C(i) curves) decreased only in YP foliage exposed to 2 x CO(2)-air. In general, growth was significantly stimulated by 2 x CO(2) in all three species; though there were few significant growth responses following exposure to 2 x O(3) or the combination of 2 x CO(2) plus 2 x O(3). Results indicate that responses to interacting stressors such as O(3) and CO(2) are species specific.