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.     

Effects of SO2, NO2, and O3 on population development and morphological and physiological parameters of native herb layer species in a beech forest

The native ground vegetation of a beech forest was fumigated with moderate doses of SO(2), NO(2), and O(3) in open-top chambers. The effects of fumigants on growth and above-ground development were dependent on species. The treatments caused early senescence in several of the species present. The epicuticular waxes were attacked by fumigation, which was shown by higher wettability of the leaf surfaces and by leaching of ions. Interspecific differences were observed in the responses of transpiration and photosynthesis to fumigation. Similar patterns of effects on transpiration and photosynthesis, however, were found in the same species. Carbohydrate metabolism was altered by fumigation, leading to starch accumulation in the leaves. Besides effects on higher plants, fumigations also resulted in alterations of the soil microflora. The bacteria/fungi ratio was depressed at the fumigation plots.     

Effects of reactive hydrocarbons and hydrogen peroxide on antioxidant activity in cherry leaves

One-year-old cherry trees were fumigated with propene and gas-phase hydrogen peroxide, singly and in combination, in controlled-environment chambers for an 8-week period during the summer season. A UV light source was included with the combined propene and hydrogen peroxide regime to provide a source of hydroxyl radicals and ozone, and thus all the constituents of a photochemical smog. Measurements were made of soluble protein concentration and of glutathione reductase activity in leaf extracts from two or three leaf classes in plants from each treatment regime at the end of each fumigation period. Significant increases in soluble protein concentration with respect to the controls were found in plants fumigated with propene and hydrogen peroxide. The occurrence and extent of these differences depended on the leaf class and on the timing of the fumigation period over the summer with respect to bud break. The activity of glutathione reductase was found to be significantly increased in mature lower leaves of plants which had been fumigated with hydrogen peroxide. This effect was independent of the timing of fumigation with respect to bud break. Enzyme activity was also increased in propene and in propene plus hydrogen peroxide treatments, but only when plants were fumigated early in the growth season.     

Persistent stimulation of CO2 assimilation and stomatal conductance by summer ozone fumigation in Norway spruce

CO(2) assimilation rate, stomatal conductance and chlorophyll content of current and previous years' needles of Norway spruce were measured in May 1988, 205 days after the cessation of ozone fumigation during the summer of 1987. Rates of assimilation were consistently higher for both needle year age classes for ozone fumigated trees in comparison to control trees, although only statistically significant for part of the day for current year's needles. A 26% and 48% stimulation, overall, in mean daily rates of assimilation for current and previous years' needles of ozone fumigated trees was observed. This was due to an enhanced apparent quantum yield and light saturated rate of assimilation of ozone fumigated trees. The temperature response regression of assimilation versus temperature was also greater, such that at any given temperature, assimilation was higher for ozone treated trees than control trees. Stomatal conductance was greater for ozone fumigated trees than the controls, but this was only marginally statistically significant. Moreover, there was a consistent increase in chlorophyll content in both year classes in ozone-treated trees. These results are discussed in relation to a possible long term effect of ozone fumigation upon the processes of conifer winter hardening and spring de-hardening.     

Effects of sulfur dioxide on growth, photosynthesis and enzyme activities of Chinese Guger-Tree seedlings

The Chinese Guger-Tree (Schima superba Gard et Champ var. superba) is an important harwood species in Taiwan where the ambient SO(2) concentration in some areas is high. Seedlings were raised in field chambers with and without SO(2) to determine whether this species is affected by this pollutant. After 4 weeks of exposure to 325 ppb of SO(2), the photosynthetic rate of seedlings decreased immediately. During the fumigation period, stem height growth was not inhibited, however, the stem diameter growth was reduced significantly. The dry weight of leaves was unchanged, while stem, root and total seedling weight were lower than those of control plants. Sulfhydryl groups in leaves increased by 75%, whereas they did not change in roots following SO(2) uptake. Superoxide dismutase in leaves did not change, however, peroxidase activity increased significantly. Results suggest that ambient SO(2) in some areas in Taiwan may affect the physiology and growth of the Chinese Guger-Tree.     

Influence of ozone air pollution on plant-herbivore interactions. Part 2: Effects of ozone on feeding preference, growth and consumption rates of monarch butterflies (Danaus plexippus)

Effects of ozone fumigation of Asclepias curassavica L. and A. syriaca L. on feeding preference, growth, development, and nutritional indices of monarch larvae were investigated in conjunction with changes in specific leaf metabolites. While foliar chemistry was quite variable, fumigation generally decreased sugars and proteins, and increased amino acids and phenolics in A. curassavica. Effects were similar in A. syriaca except that sugars were generally increased while amino acids were usually not affected. On A. curassavica, 3rd instar larvae preferred ozone-treated leaves while 4th instars showed no preference; conversely, on A. syriaca, 3rd instars showed no preference while 4th instars preferred control leaves. Relative growth rate and relative consumption rate of 5th instars were greater on fumigated plants of both species, but other nutritional indices were unaffected. Larvae developed more rapidly on intact fumigated plants of both species than on the respective controls. The results suggest that enhanced feeding stimulation may be the primary cause of the altered behavior and performance on ozone-fumigated plants.     

Responses to ozone of insects feeding on a crop and a weed species

The influence of ozone on insect herbivore growth and population development was investigated. Fumigation of both pea (Pisum sativum L.) and dock (Rumex obtusifolius L.) at a range of O(3) concentrations between 21-206 nl litre(-1) produced changes in mean relative growth rates of the aphids Acyrthosiphon pisum Harris and Aphis rumicis L. of between 24 and -6% relative to controls. However, there was no evidence of a dose-related response to O(3) fumigation and no clear differences in aphid response when fumigated with the plant on prefumigated or previously unfumigated plant material. It is suggested that this may, in part, be due to the presence of NO contamination during O(3) fumigation. However, the MRGR of dock aphids was found to be greater on new compared to old leaves as well as the increase on the new growth and decrease on the old growth of fumigated plants relative to unfumigated controls. The size of egg batches of the chrysomelid beetle Gastrophysa viridula Degeer were found to be larger, survival and productivity of larvae was higher, and the food consumption lower on R. obtusifolius fumigated with 70 nl litre(-1) O(3) compared with unfumigated controls. This meant that these beetle larvae consumed less leaf area per mg of production on fumigated leaves probably because of their better nutritional quality and/or reduced leaf defences. However, the rate of development of larvae was similar on fumigated and control plants.     

Secondary effects of air pollution: ozone decreases brown rust disease potential in wheat

Young wheat plants were fumigated with 170 microg m(-3) ozone for 3 days, or with 210 microg m(-3) ozone for 7 days, for 7 hours a day. At the end of the fumigation period the plants were inoculated with brown rust (Puccinia recondita f. sp. tritici) uredospores. The development of new uredospore pustules on fumigated and control plants was evaluated as a measure of rust disease potential. The number of pustules on the ozone fumigated plants was greatly reduced in comparison with the number of plants treated with charcoal-filtered air.     

Effects of simultaneous ozone exposure and nitrogen loads on carbohydrate concentrations, biomass, growth, and nutrient concentrations of young beech trees (Fagus sylvatica)

Beech seedlings were grown under different nitrogen fertilisation regimes (0, 20, 40, and 80 kg Nha(-1)yr(-1)) for three years and were fumigated with either charcoal-filtered (F) or ambient air (O3). Nitrogen fertilisation increased leaf necroses, aphid infestations, and nutrient ratios in the leaves (N:P and N:K), as a result of decreased phosphorus and potassium concentrations. For plant growth, biomass accumulation, and starch concentrations, a positive nitrogen effect was found, but only for fertilisations of up to 40 kg Nha(-1) yr(-1). The highest nitrogen load, however, reduced leaf area, leaf water content, growth, biomass accumulation, and starch concentrations, whereas soluble carbohydrate concentrations were enhanced. The ozone fumigation resulted in reduced leaf area, leaf water content, shoot growth, root biomass accumulation, and decreased starch, phosphorus, and potassium concentrations, increasing the N:P and N:K ratios. A combined effect of the two pollutants was detected for the leaf area and the shoot elongation, where ozone fumigation amplified the nitrogen effects.     

Seasonal changes in the tissue-metal (Cd, Zn and Pb) concentrations in two ecophysiologically dissimilar earthworm species: pollution-monitoring implications

During two consecutive growing seasons, the same potted individuals of European aspen (Populus tremula), grown from root cuttings of one clone, were fumigated with either ambient air or ozone concentrations of 0 (control), 0.05 or 0.1 microlitre litre(-1). Structure and biomass of the annually formed branches were analysed after excision at the end of each season. Only at 0.1 microlitre litre(-1) was branch weight reduced, and crooked axes occurred in each season. During the second season, branch length and leaf sizes were strongly reduced, while many leaves displayed yellowish deficiency symptoms and lowered cation concentrations. Such leaves contrasted to those showing characteristic O3-bronzing. Although foliage density was enhanced due to reduced branch length, the area of attached foliage was limited by the small leaf sizes, necrotic leaves and premature leaf loss. During mid-summer of the second fumigation period, photosynthetic capacity, carboxylation efficiency and water-use efficiency (WUE) declined in (attached) yellowish and bronze leaves at 0.1 microlitre litre(-1), whereas green leaves at 0.05 microlitre litre(-1) displayed accelerated senescence in late summer while maintaining WUE. It is concluded that the differences in branch growth between the two growing seasons were caused in part by internal changes in those plant organs (root and basal stem), which had experienced both fumigation periods.     

Effect of chronic ozone fumigation on the photosynthetic process of poplar clones showing different sensitivity.

Rooted cuttings from two poplar clones (Populus x euramericana, I-214, and Populus deltoides x maximowiczii, Eridano) were exposed for 15 days to diurnal square-wave treatment with ozone (60 nL L-1 for 5 h day-1). Completely fully expanded leaves exposed to ozone showed a reduction in net CO2 assimilation rate as compared to the control leaves during whole exposure period in both the clones. The reduction was related to a strong stomatal closure in clone I-214, but also to an altered mesophyll activity ascribed to limitation of the dark reactions of photosynthetic process. The results obtained in leaves of I-214 subjected to long-term fumigation seem to support the view that the decrease in quantum yield of electron transport may be a mechanism to down-regulate photosynthetic electron transport so that production of ATP and NADPH would be in equilibrium with the decreased demand in the Calvin cycle. In Eridano the CO2 assimilation was reduced because of the exposure and any alteration in stomatal conductance was observed. Thus, chlorophyll fluorescence parameters showed that an inhibition of photosystem II had occurred (reduction in Fv/Fm ratio), while no alterations in quenching parameters were observed upon illumination. The results seem to indicate that an alternative sink for reducing equivalent, other than carbon metabolism is present.     

Tree age dependence and within-canopy variation of leaf gas exchange and antioxidative defence in Fagus sylvatica under experimental free-air ozone exposure

We characterized leaf gas exchange and antioxidative defence of two-year-old seedlings and 60-year-old trees of Fagus sylvatica exposed to ambient (1 x O3) or two-fold ambient (2 x O3) O3 concentrations (maximum of 150 ppb) in a free-air canopy exposure system throughout the growing season. Decline in photosynthesis from sun-exposed to shaded conditions was more pronounced in adult than juvenile trees. Seedling leaves and leaves in the sun-exposed canopy had higher stomatal conductance and higher internal CO2 concentrations relative to leaves of adult trees and leaves in shaded conditions. There was a weak overall depression of photosynthesis in the 2 x O3 variants across age classes and canopy positions. Pigment and tocopherol concentrations of leaves were significantly affected by canopy position and tree age, whereas differences between 1 x O3 and 2 x O3 regimes were not observed. Glutathione concentrations were significantly increased under 2 x O3 across both age classes and canopy levels. Seedlings differed from adult trees in relevant physiological and biochemical traits in ozone response. The water-soluble antioxidative systems responded most sensitively to 2 x O3 without regard of tree age or canopy position.     

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 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.     

Variation in ozone sensitivity among clones of Betula pendula and Betula pubescens

Forty clones of Betula pendula and 6 clones of Betula pubescens, originating from southern and central Finland, were ranked in order of ozone sensitivity according to visible injuries, growth and leaf senescense under low ozone exposure. The plants were fumigated in natural climatic conditions using an open-air exposure system during two growing seasons. Control plants were grown under ambient air, and the elevated-ozone exposures were 1.6x the ambient in 1994 and 1.7x the ambient in 1995. The differences in ozone sensitivity among clones were large. Ozone tolerance was related to thicker leaves and higher stomatal density as compared to sensitive clones. Ultrastructural ozone-induced symptoms were found in chloroplasts of sensitive clones. Increased number of visibly injured leaves on fumigated plants was correlated with reduced leaf formation, foliage area, shoot dry wt and number of stomata, and increased yellowing of leaves. The results suggest that a considerable proportion of birch trees, showing high sensitivity to ozone, are at risk if ambient ozone exposures increase.     

Ozone fumigation (twice ambient) reduces leaf infestation following natural and artificial inoculation by the endophytic fungus Apiognomonia errabunda of adult European beech trees

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

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.     

CO2/H2O gas exchange parameters of one- and two-year-old needles of spruce and fir

Gas exchange was characterized in one- and two year-old spruce (Picea abies L. Karst.) and fir seedlings (Abies alba Mill.) which had been exposed to low levels of ozone, sulfur dioxide and simulated rain or a combination of all three variables in open top chambers from 1983 through 1988. The gas exchange measurements were carried out in March 1988 at the end of the five year experiment. The twigs examined did not exhibit any visible sign of injury, specifically no differences were apparent between trees under the treatments of simulated acidic rain at pH 5.0 and pH 4.0. The study of carbon dioxide response curves showed different effects of the pollutants on the tree species. One-Year-old spruce needles treated with O(3) and simulated acidic precipitation pH 4.0 showed noticeable reduction of net photosynthetic rate. Exposure to the combination O(3) and SO(2) at pH 4.0 resulted in a significant depression of photosynthesis in two-year-old needles Transpiration rate was not decreased to a similar extent. No changes either in photosynthesis or transpiration were found in spruce under fumigation with SO(2) alone. These results indicate that ozone is the principal cause of changes in photosynthetic performance of spruce. It alters mesophyll response rather than reducing stomatal conductance. The specific changes that occur in the mesophyll could be diagnosed as inactivation of a carbon fixing enzyme as well as damage of the electron transport system. Fir seem to be more tolerant to ozone. No changes in photosynthesis and transpiration following exposure to O(3) alone were found. However, SO(2) fumigation, alone or in combination with O(3), resulted in a marked decrease of photosynthetic performance. Particularly, carboxylation efficiency and also maximum carboxylation velocity were depressed indicating a reduction in carbon fixing enzyme activity. No differences between single and combined fumigation treatments regarding these variables were determined. However, parameters measured to determine changes in electron transport rate showed a higher depression in the presence of both pollutants. Transpiration also was reduced by SO(2).     

Factors Affecting Ozone Sensitivity and Susceptibility Of Cotton Plants

Cotton plants were investigated to ascertain the growth period of maximum sensitivity to ozone. Plants were grown from seed in the greenhouse to obtain a two-day interval age series to about three months old. Before fumigation with 0.2 to 1.0 ppm ozone for 1 hr, plants were preconditioned under standard growth chamber conditions with day temperatures of 32°, night temperatures of 21°, relative humidity of 50%, and a light intensity of 6 X 10⁴ ergs/cm²/sec. Also, stomatal opening was monitored with a resistance hygrometer. Ozone damage to leaves was assessed by rating per cent visible damage with an arbitrary scale. Approximately 0.6-0.7 ppm ozone exposure for 1 hr was necessary for visible damage. Leaves were susceptible to ozone when about three-quarters fully expanded. Leaves appearing on older plants became progressively less sensitive. Diurnally, susceptible leaves were only sensitive after several hours in the light and then lost sensitivity toward the end of a normal day. The two most critical questions concerning the nature of ozone sensitivity and susceptibility are: (7) what is the nature of susceptibility, and (2) what is the nature of sensitivity of susceptible leaves?     

Atmospheric flux of agricultural fumigants from raised-bed,plastic-mulch crop production systems

Atmospheric emission of methyl isothiocyanate (MITC), chloropicrin (CP), 1,3-dichloropropene (1,3-D), and dimethyl disulfide (DMDS) were measured in the field under fumigant application scenarios representative of raised bed–plastic-mulched crop production systems. For three fumigation sites located in Florida, cumulative emissions of 1,3-D, MITC and CP were less than 11%, 6% and 2%, respectively. For three fumigation sites in located in Georgia, cumulative emissions of MITC and CP were <13% and 12%, respectively while DMDS emissions varied from 37% to 95%. In the Florida sites, emission peak flux of CP occurred within the first 6 h after application. Peak emission of 1,3-D and MITC occurred between 100 and 144 h after application. In the Georgia sites where fumigated soil was covered by low density polyethylene (LDPE) plastic, emission peak flux of DMDS and MITC occurred between 12and 48 h after application. Key factors affecting atmospheric emissions were soil moisture, soil tilth and the resistance to fumigant diffusion of the plastic film used to cover soil following application. This study demonstrated reduced atmospheric emissions of agricultural fumigants under commercial production conditions when applied using good agricultural practices including soil water contents above field capacity, uniform soil tilth in the fumigation zone and the use of metalized or virtually impermeable films to further reduce fumigant emissions. The results of this study show a need for regional flux studies due to the various interactions of soil and climate with local agricultural land management practices.