Reactions of cloned poplars to air pollution

Ozone-induced changes in ethylene production, ACC oxidase activity and the contents of ACC, MACC and free PAs were studied inPopulus nigra L. cv. Loenen with high ozone sensitivity as judged by the degree of chlorophyll degradation and premature leaf abscission. Ethylene production, ACC oxidase activity, ACC content and MACC levels were induced by the one-, two-, and three-week ozone exposure (36±9 ppb O₃ for 11 hours a day). In addition, increases in PA levels, especially in spermidine, were measured in ozone treated plants. The role of free PAs and MACC synthesis as possible antisenescence reactions is discussed.     

Growth-stage dependent crop yield response to ozone exposure

Data from four crop yield-loss field trials were examined to determine if analysis using an imposed phenological weighting function based on seasonal growth stage would provide a more accurate indication of impact of ozone exposure. Alfalfa (Medicago sativa L. cv. Moapa 69), dry bean (Phaseolus vulgaris L. cv. California Dark Red kidney), fresh market and processing tomato (Lycopersicon esculentum Mill. cv. 6718 VF and VF-145-B7879, respectively) were grown at 9-11 ambient field plots within southern California comprising an ambient gradient of ozone. The growing season for each crop was artificially divided into 'quarters' composed of equal numbers of whole days and roughly corresponding to specific growth stages. Ozone exposure was calculated for each of these 'quarters' and regressed against final crop yield using 163 different exposure statistics. Weighting functions were developed using reciprocal residual mean square (1/RMS) or percentage of the best 100 exposure statistics of the 163 tested (TOP100) for each of the quarters. The third quarter of the alfalfa season was clearly most responsive to ozone as measured by both of the weighting functions. Third quarter ozone was also weighted highest by both weighting functions for dry bean. Fresh market and processing tomato were each influenced the greatest by second quartero zone as demonstrated by both weighting functions. The occurrence of ozone during physiologically important events (flowering and initial fruit set in second quarter for tomato; pod development in third quarter for dry bean) appeared to influence the yield of these crops the greatest. Growth-stage-dependent phenological weighting of pollutant exposure may result in more effective predictions of levels of ozone exposure resulting in yield reductions.     

Chronic effects of water-borne PFOS exposure on growth, survival and hepatotoxicity in zebrafish: a partial life-cycle test

Perfluorooctane sulfonate (PFOS) is widely distributed and persistent in the environment and wildlife. The main aim of this study was to investigate the impact of long-term exposure to low concentrations of PFOS in zebrafish. Zebrafish fry (F(0), 14 d post-fertilization, dpf) were exposed via the water for 70 d to 0 (control), 10, 50 and 250 microg L(-1) PFOS, followed by a further 30 d to assess recovery in clean water. The effects on survival and growth parameters and liver histopathology were assessed. Although growth suppression (weight and length) was observed in fish treated with high concentrations PFOS during the exposure period, no mortality was observed throughout the 70 d experiment. Embryos and larvae (F(1)) derived from maternal exposure suffered malformation and mortality. Exposure to 50 and 250 microg L(-1) PFOS could inhibit the growth of the gonads (GSI) in the female zebrafish. Histopathological alterations, primary with lipid droplets accumulation, were most prominently seen in the liver of males and the changes were not reversible, even after the fish were allowed to recover for 30 d in clean water. The triiodothyronine (T(3)) levels were not significantly changed in any of the exposure groups. Hepatic vitellogenin (VTG) gene expression was significantly up-regulated in both male and female zebrafish, but the sex ratio was not altered. The overall results suggested that lower concentrations of PFOS in maternal exposure could result in offspring deformation and mortality.     

Evaluation of fluoranthene phytotoxicity in pea plants by Hill reaction and chlorophyll fluorescence

The effect of both increased concentrations (0.01 and 1 mg l(-1)) of fluoranthene (FLT) and the duration of exposure (18 and 25 days) on the growth and photosynthetic processes in pea plants (Pisum sativum L., cv. Garde) was investigated. FLT concentration in roots and shoot of pea plants was also determined. The obtained results demonstrated that the higher concentration of FLT (1 mg l(-1)) significantly inhibited the growth of the pea plants after 25 days of the application, also affected the content of photosynthetic pigments (chlorophyll a, b and carotenoids), and the primary photochemical processes of photosynthesis. In chlorophyll fluorescence parameters, the significant increase of F(0) values and the decrease of F(V)/F(M) and Phi(II) values was recorded. The Hill reaction of isolated chloroplasts of pea plants was significantly inhibited after 25 days by presence of FLT (0.01 and 1 mg l(-1)) in nutrient solution, while after 18 days no significant response of Hill reaction activity was recorded. The fluoranthene content in roots and shoot of pea plants increased with increasing FLT concentration in the environment and the substantial accumulation of FLT was observed in the roots.     

Photosynthetic inhibition and superoxide dismutase activity in soybean cultivars exposed to short-term ozone fumigations

Net photosynthesis (Pn) and mean stomatal conductance (gs) of soybean (Glycine max (M.) L.) cvs 'Dare', 'Essex' and 'Williams 82' were inhibited by exposure to 200 nliters liter(-1) ozone (O(3)) for 4 h. Changes in gas exchange rates were not uniform over time, and a near steady state condition was attained after fumigation for 2 h. There was no recovery of Pn or gs for up to 2 h following termination of O(3) application. The cv 'Dare' was considered most sensitive to O(3) since, in this cultivar, gs was least responsive during the initial 2.5 h of fumigation and the estimated internal O(3) flux was about 10% greater than in the other cultivars. Polyacrylamide gel electrophoresis of soybean leaf extracts, followed by activity staining for superoxide dismutase (SOD), revealed the presence of numerous major and minor bands. Cynide sensitivity of several isoforms indicated the presence of both Cu-Zn and Mn SODs. In cv 'Essex', each metalloform accounted for 50% of the total SOD activity. No qualitative differences in SOD banding patterns were evident among the three soybean cultivars before or after O(3) treatment. Ozone fumigation for 4 h did not significantly alter SOD activity in any cultivar. The contribution of gs, as a flux limitation, and SOD activity are discussed as tolerance mechanisms to O(3) in soybean.     

Yield responses of different crop species to long-term fumigation with sulphur dioxide in open-top chambers

Potted plants of commercial cultivars of rape (Brassica napus L., cv. 'callypso'), summer barley (Hordeum vulgare L., cvs. 'arena' and 'hockey') and bush beans (Phaseolus vulgaris L., cvs. 'rintintin' and 'rosisty') were continuously exposed in open-top chambers to sulphur dioxide (SO(2)) for the whole growing season in order to assess effects of this pollutant on growth and various yield parameters. Treatments consisted of charcoal-filtered air (CF) and CF supplemented with four levels of SO(2), resulting in mean exposure concentrations (microg m(-3)) of approximately 8, 50, 90, 140 and 190. With the exception of the 1000 seeds weight, which was slightly reduced, dry matter production and yield parameters of rape remained unaffected by all SO(2) concentrations or were even stimulated. Compared to CF vegetative growth of both bean cultivars was reduced by 10-26% at all SO(2) levels; however, with significant effects only for cv. 'rintintin'. While all SO(2) additions reduced significantly the yield (dry weight of pods) of the bean cultivar 'rosisty' between 17% and 32%, cv. 'rintintin' showed a significant reduction of up to 42% only at the two highest pollutant concentrations. Dry matter production of the barley cultivars was mainly impaired at SO(2) concentrations > 100 microg m(-3) with a reduction of 30-52%. While nearly all yield parameters of cv. 'hockey' reacted similar to the dry matter production, the yield of cv. 'arena' was reduced already at the low SO(2) levels. At a treatment concentration of 90 microg SO(2) m(-3) a significant yield loss of 30% was recorded. A reduction of the 1000 grains weight mainly contributed to these yield losses observed for both barley cultivars. From these results, it may be assumed that SO(2) concentrations within the range 50-90 microg m(-3) are potentially phytotoxic to some crop species.     

Population dynamics in Daphnia magna as modified by chronic tetradifon stress

Two Daphnia magna offsprings (animals from the first and third brood) whose parentals (F0-generation) were exposed during 21 days to different tetradifon (4-chlorophenyl 2,4,5-trichlophenyl sulfone) concentrations, were transferred to a pesticide free medium during 21 days (recovery period). The algae Nannochloris oculata (5 x 10(5) cells/mL) was used as food. In this recovery study, survival, growth and reproduction (mean total young per female, mean brood size, onset of reproduction and mean number broods per female) were assessed as individual parameters and the intrinsic rate of natural increase (r) as population parameter, for F1 generation (1st and 3rd broods). Reproduction was still reduced in F1 (1st and 3rd broods) generation daphnids from parentals (F0) exposed to 0.18 mg/L tetradifon and higher concentrations even after 21 days in clean water. However, survival was not significantly different (p > 0.05) in those F1 offsprings from parentals pre-exposed to the acaricide. Growth was still reduced in F1 daphnids from parentals pre-exposed to 0.10-0.44 mg/L tetradifon. The intrinsic rate of natural increase (r) was still affected in F1 generation daphnids, specially in those from the third brood. Therefore, F1 generation D. magna offsprings from a parental generation (F0) previously exposed to the acaricide tetradifon were not able to recuperate completely when a recovery period of 21 days was allowed.     

Effects of ethylenediurea and ozone on the antioxidative systems in beans (Phaseolus vulgaris L.)

To study the biochemical mechanism of EDU protection against ozone injury, peroxidase, ascorbate-dependent peroxidase, and catalase activities, and the contents of ascorbic acid, dehydroascorbic acid, malondialdehyde and soluble protein were measured in Phaseolus vulgaris L. cv. Lit exposed to ozone and ethylenediurea (EDU) in open-top chambers. Plants not treated with EDU showed foliar bronzing due to ozone, while EDU-treated plants were not affected. EDU application modified the reaction of biochemical parameters to ozone. Soluble protein content was elevated by EDU. Peroxidase activity increased with ozone exposure in untreated plants only, while ascorbate-dependent peroxidase activity was lower in EDU treated plants. Catalase activity decreased in EDU-untreated plants. The ratio of ascorbic acid to dehydroascorbic acid was significantly increased in EDU treated plants. These results suggest that EDU might induce ascorbic acid synthesis and therefore provide the plant with a very potent antioxidant. Or the content of hydrogen peroxide was reduced due to other unknown processes and caused a delay in foliar senescence, regardless of whether these processes were ozone-induced or due to natural aging processes.     

Effects of ozone exposure in open-top chambers on poplar (Populus nigra) and beech (Fagus sylvatica): a comparison

Rooted cuttings of poplar (Populus nigra) and seedlings of beech (Fagus sylvatica) were exposed to ozone in open-top chambers for one growing season. Three treatments were applied: charcoal-filtered (CF), non-filtered (NF) and non-filtered air plus 30 ppb (nl l(-1)) ozone (NF+). Extra ozone was only added on clear days, from 09:00 until 17:00-20:00. The AOT40s (accumulated exposure over a threshold of 40 ppb), calculated from April to September were 4055 ppb.h for the NF and 8880 ppb.h for the NF+ treatments. For poplar ozone exposure caused highly significant reductions in growth rate, light-saturated net CO(2) assimilation rate, stomatal conductance, F(v)/F(m) and chlorophyll content. The largest effects were observed in August at which time ozone concentrations were elevated. A reduction was noticed in new leaf production, while accelerated ageing and visible damage to leaves caused high leaf losses. For beech the responses were similar but less pronounced: ozone exposure resulted in non-significant growth reductions, slight changes in light-saturated photosynthesis and accelerated leaf abscission. The chlorophyll content of beech leaves was not affected by the ozone treatments. The results confirmed previous observations that fast-growing tree species, such as most poplar species and hybrids, are more sensitive and responsive to tropospheric ozone than slower-growing species, such as beech. The growth reductions observed and reported here for beech were within the range of those reported in relationship to the AOT40 (accumulated exposure over a threshold of 40 ppb) critical level for ozone.     

Ozone differentially affects physiological and biochemical responses of two clover species; Trifolium repens and Trifolium pratense

The effect of acute ozone exposure (150 ppb for 3 h) on two clover species, white clover (Trifolium repens L.) and red clover (Trifolium pratense L.) was investigated through the analysis of 10 different physiological and biochemical parameters. Twenty-four hours after fumigation, visible symptoms of injury on leaves were observed only in red clover, but from the biochemical point of view, both species revealed significant ozone-induced modifications. A decrease in the photosynthetic efficiency as well as an increase in the de-epoxidation index and a decrease in the redox state of ascorbate were detected only in T. pratense leaves; no significant change in pigment content was found in either of the two species. On the other hand, both white and red clover showed, although to different extents, significant decreases in the activities of the antioxidant enzymes peroxidase and ascorbate peroxidase. Multivariate statistical analysis revealed not only that ozone affects both species, but also that they differentially respond to the pollutant, confirming the higher sensitivity of Trifolium pratense to ozone exposure.     

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.     

Influence of ozone air pollution on plant-herbivore interactions. Part 1: Biochemical changes in ornamental milkweed (Asclepias curassavica L.; asclepiadaceae) induced by ozone

A series of fumigation experiments was conducted with bloodflower (Asclepias curassavica L.) in continuous-flow stirred reactors (CSTRs) to elucidate the effects of ozone on foliar concentrations of several primary and secondary plant metabolites relevant to herbivores. Plants 8 weeks of age were subjected to different ozone levels ranging from 0 to 134 nl liter(-1) for exposure periods up to 16 days. Leaves were analyzed for concentration of soluble carbohydrates, starch, free amino acids, soluble protein, total phenolics, and total cardenolides. Significant interactions between the linear effects of ozone concentration and exposure time were found for soluble carbohydrates, amino acids, cardenolides and phenolics. No significant treatment effects could be observed on foliar starch and protein concentration. The metabolic responses of plants to fumigation appeared to be altered by overall plant nutrition. It is possible that the metabolic changes observed in the host plant represent important changes in nutritional quality to insects.     

Removal of pyrene and benzo(a)pyrene from contaminated water by sequential and simultaneous ozonation and biotreatment.

The removal of pyrene and benzo(a)pyrene from contaminated water by sequential and simultaneous ozonation-bioremediation techniques was investigated. During the sequential treatment, ozonation using 0.5 or 2.5 mg/L ozone was used as a pretreatment process, whereas, during the simultaneous treatment process, ozonation of hydrocarbon-contaminated water at a predetermined duration using 0.5 mg/L ozone was made in the presence of microbial biomass. Ozonation was not beneficial for the removal of pyrene. However, despite a decreased specific biodegradation rate, ozonation improved the overall elimination of benzo(a)pyrene during both treatment processes. The overall removal of benzo(a)pyrene increased from 23 to 91% after exposure of the water to 0.5 mg/L ozone for 30 minutes during the simultaneous treatment process and further to 100% following exposure to 2.5 mg/L ozone for 60 minutes during the sequential treatment mode, demonstrating the benefits of combined ozonation-biological treatment for the removal of polycyclic aromatic hydrocarbons.     

Growth and yield responses of spring wheat (Triticum aestivum L. CV. Turbo) grown in open-top chambers to ozone and water stress

Spring wheat (Triticum aestivum L.) cv. Turbo was exposed to different levels of ozone and water supply in open-top chambers in 1991. The plants were grown either in charcoal filtered air (CF), not filtered air (NF), in charcoal filtered air with proportional addition of ambient ozone (CF1), or in charcoal filtered air with twice proportional addition of ambient ozone (CF2). The mean seasonal ozone concentrations (24 h mean) were 2.3, 20.6, 17.3, and 24.5 nl litre(-1) for CF, NF, CF1, and CF2 treatments, respectively. Ozone enhanced senescence and reduced growth and yield of the wheat plants. At final harvest, dry weight reductions were mainly due to reductions in ear weight. Grain yield loss by ozone mainly resulted from depressions of 1000 grain weight, whereas numbers of ears per plant and of grains per ear remained unchanged. Pollutants other than ozone did not alter the response to ozone, as was obvious from comparisons between CF1 and NF responses. Water stress alone did not enhance senescence, but also reduced growth and yield. However, yield loss mainly resulted from reductions in the number of ears per plant; 1000 grain weight was not influenced by water stress. No water supply by ozone treatment interactions were detected for any of the estimated parameters.     

Response of abscisic acid in soybean leaves to NO2 exposure

Soybean (Glycine max L. Merril) cv. 'Williams' plants were exposed 0.0, 0.05, 0.2 and 0.4 microl litre(-1) NO(2) under controlled environment. Amount of abscisic acid (ABA) was measured after either 7 h or 5 days of exposure as well as after 18 h of recovery period. For quantitative analysis of ABA enzyme-linked immunosorbent assay was used. A significant increase (160%) was observed in plants treated with 0.4 microl litre(-1) NO(2) for 5 days compared to an increase of only 82% on exposure for 7 h. Rate of recovery for plants treated with low doses of NO(2) was higher than those treated with higher doses.     

Ozone-induced changes in CO2 assimilation, O2 evolution and chlorophyll a fluorescence transients in barley

Spring barley (Hordeum vulgare cv. Klaxon) plants, 9 days old, were exposed to 0.05, 0.10 or 0.15 microl litre(-1) ozone (O3) for 12 days. Fumigation was administered for 7 h between 9.00 h and 16.00 h each day. Using conventional IRGA equipment, the carbon dioxide exchange rate (CER) was shown to decrease with increasing concentration of O3 during the exposure period, falling to 60% of the control value at the highest O3 concentration. Transpiration rates and stomatal conductance showed similar trends. Light saturation curves, obtained using a leaf disc oxygen electrode, demonstrated that O3-treated leaves had lower apparent quantum yields (QY) and generally lower rates of O2 evolution at saturating light and CO2 levels. Oscillations in chlorophyll a fluorescence, normally observed in control plants, could not be detected after O3 treatment and could only be restored to some extent by feeding the phosphate sequestering agent D-mannose to the leaves.     

The effects of ozone-exposed sugar maple seedlings on the biological performance and the feeding preference of the forest tent caterpillar (Malacosoma disstria Hbn.)

The effects of exposure of sugar maple (Acer saccharum Marsh.) to ozone on the entire larval stage of a native insect have not been previously investigated. This study reports the effects of sugar maple seedlings exposed to different ozone concentrations on the relative performance and the feeding preference of the forest tent caterpillar (Malacosoma disstria Hbn.). Three-year-old seedlings were set in nine open-top field chambers in the spring of 1992 and 1993. Three ozone concentrations were generated: charcoal-filtered ambient air (0x), ambient air (1x) and three times ambient air (3x). In 1992, female and male larval development time did not differ among ozone treatments. In 1993, female larvae reared on 3x developed faster than those on 0x and 1x, while male larvae were not affected. Ozone treatments did not influence pupal weights except for males in 1993 where pupae reared on 0x were heavier than 1x but did not differ from 3x. Larval and pupal survival rates were not affected by ozone in either year. Finally, 4th and 5th instar larvae showed a significant feeding preference for 3x foliage in 1993 but not in 1992. The response of the forest tent caterpillar to ozone exposed seedlings varied between years and could be more sensitive to annual climatic variations than ozone.     

Chlorophyll a fluorescence, antioxidant enzymes and lipid peroxidation in tomato in response to ozone and benomyl.

Ozone is a widely distributed phytotoxic air pollutant and is known to reduce the yield of several important agricultural crops in Spain. However, benomyl has been found to lessen the adverse impact of ozone on plants. We studied the effects of ozone and benomyl on chlorophyll a fluorescence, antioxidant enzymes, and lipid peroxidation in tomato (Lycopersicon esculentum Mill. cv. Tiny Tim) grown in open-top chambers in the field. Our results indicate that benomyl prevented the peroxidation of membrane lipids and increased protection of PSII from ozone. There was also a significant reduction in the activity of the antioxidant enzyme superoxide dismutase in ozone-exposed plants that had not been treated with benomyl. Comparing plants treated with benomyl to untreated plants we found that, on exposure to ozone, a greater fraction of light absorption energy was cycled through the photosynthetic system in benomyl-treated plants, as shown by the higher PSII-mediated electron flow and the higher fraction of open PSII reaction centers. The values analyzed in the fluorescence parameters and lipid peroxidation were similar for plants without benomyl grown in a charcoal-filtered environment and benomyl-treated plants exposed to ozone.     

Effects of sublethal exposure to a pesticide on levels of energetic compounds in Anguilla anguilla.

Abstract

The effect of two sublethal fenitrothion concentrations (0.02 and 0.04 mg/L) on the energy metabolism of the european eel Anguilla anguilla and its recovery from intoxication was investigated. Analysis of various parameters such as glycogen, proteins and total lipids was made on liver and muscle eel tissues after 2, 8, 12, 24, 32, 48, 56, 72 and 96 hr of fenitrothion (0,0‐dimethyl 0–3‐methyl‐4‐nitrophenyl phosphorothioate) exposure. Subsequently, the fish were allowed recovery periods of 8, 12, 24, 48, 72, 96, 144 and 192 hr in clean water, and the same parameters were again evaluated. The results obtained during the exposure to the pesticide as well as during the recovery phase were used to calculate the caloric content in both tissues of A. anguilla. A reduction in energy reserves in the selected tissues was observed after exposure to both fenitrothion concentrations and the caloric content in those animals was lower than in the controls. Most of the metabolic disorders did not persist after allowing recovery in clean water during a week.