Ethylenediurea (EDU): a research tool for assessment and verification of the effects of ground level ozone on plants under natural conditions

Ethylenediurea (EDU) has been widely used to prevent ozone (O₃) injury and crop losses in crop plants and growth reductions in forest trees. Successful use requires establishing a dose/response curve for EDU and the proposed plant in the absence of O₃ and in the presence of O₃ before initiating multiple applications to prevent O₃ injury. EDU can be used to verify foliar O₃ symptoms in the field, and to screen plants for sensitivity to O₃ under ambient conditions. Despite considerable research, the mode of action of EDU remains elusive. Additional research on the mode of action of EDU in suppressing O₃ injury in plants may also be helpful in understanding the mode of action of O₃ in causing injury in plants.     

Protection of plants from ambient ozone by applications of ethylenediurea (EDU): A meta-analytic review

A meta-analysis was conducted to quantitatively assess the effects of ethylenediurea (EDU) on ozone (O₃) injury, growth, physiology and productivity of plants grown in ambient air conditions. Results indicated that EDU significantly reduced O₃-caused visible injury by 76%, and increased photosynthetic rate by 8%, above-ground biomass by 7% and crop yield by 15% in comparison with non-EDU treated plants, suggesting that ozone reduces growth and yield under current ambient conditions. EDU significantly ameliorated the biomass and yield of crops and grasses, but had no significant effect on tree growth with an exception of stem diameter. EDU applied as a soil drench at a concentration of 200-400 mg/L has the highest positive effect on crops grown in the field. Long-term research on full-grown tree species is needed. In conclusion, EDU is a powerful tool for assessing effects of ambient [O₃] on vegetation.     

Use of the antiozonant ethylenediurea (EDU) in Italy: verification of the effects of ambient ozone on crop plants and trees and investigation of EDU's mode of action

Twenty-four experiments where EDU was used to protect plants from ozone (O₃) in Italy are reviewed. Doses of 150 and 450 ppm EDU at 2-3 week intervals were successfully applied to alleviate O₃-caused visible injury and growth reductions in crop and forest species respectively. EDU was mainly applied as soil drench to crops and by stem injection or infusion into trees. Visible injury was delayed and reduced but not completely. In investigations on mode of action, EDU was quickly (<2 h) uptaken and translocated to the leaf apoplast where it persisted long (>8 days), as it cannot move via phloem. EDU did not enter cells, suggesting it does not directly affect cell metabolism. EDU delayed senescence, did not affect photosynthesis and foliar nitrogen content, and stimulated antioxidant responses to O₃ exposure. Preliminary results suggest developing an effective soil application method for forest trees is warranted.     

Differential protection of ethylenediurea (EDU) against ambient ozone for five cultivars of tropical wheat

The antiozonant EDU (ethylenediurea) was used to assess the impact of ambient O₃ under field conditions on five cultivars of tropical wheat (Triticum aestivum L.). EDU solution (0 ppm and 400 ppm) was applied as soil drench (100 ml plant^?1) 10 days after germination (DAG) at an interval of 12 days. EDU-treated plants showed significant increments in stomatal conductance, photosynthetic rate, variable fluorescence, total chlorophyll, ascorbic acid, proline and protein contents and protective enzymes (POX, SOD and APX) activities in HUW468, HUW510 and HUW234 cultivars, while, a reverse trend was observed for lipid peroxidation. EDU application restored grain yield significantly by maintaining higher levels of antioxidants, metabolites and enzymes in cultivars HUW468 and HUW510. Sonalika and PBW343 showed least response of measured parameters under EDU treatment suggesting their greater resistance to O₃. EDU, thus proved its usefulness in screening suitable wheat cultivars for areas experiencing elevated concentrations of O₃.     

Protection of palak (Beta vulgaris L. var Allgreen) plants from ozone injury by ethylenediurea (EDU): Roles of biochemical and physiological variations in alleviating the adverse impacts

Ameliorative effects of ethylenediurea (N-[2-(2-oxo-1-imidazolinidyl) ethyl]-N′ phenylurea, abbreviated as EDU) against ozone stress were studied on selected growth, biochemical, physiological and yield characteristics of palak (Beta vulgaris L. var Allgreen) plants grown in field at a suburban site of Varanasi, India. Mean eight hourly ozone concentration varied from 52 to 73 ppb which was found to produce adverse impacts on plant functioning and growth characteristics. The palak plants were treated with 300 ppm EDU at 10 days after germination at 10 days interval up to the plant maturity. Lipid peroxidation in EDU treated plants declined significantly as compared to non-EDU treated ones. Significant increment in F_v/F_m ratio in EDU treated plants as compared to non-EDU treated ones was recorded. EDU treated plants showed significant increment in ascorbic acid contents and reduction in peroxidase activity as compared to non-EDU treated ones. As a result of the protection provided by EDU against ozone induced stress on biochemical and physiological characteristics of palak, the morphological parameters also responded positively. Significant increments were recorded in shoot length, number of leaves plant⁻¹, leaf area and root and shoot biomass of EDU treated plants as compared to non-EDU treated ones. Contents of Na, K, Ca, Mg and Fe were higher in EDU treated plants as compared to non-EDU treated ones. The present investigation proves the usefulness of EDU in partially ameliorating ozone injury in ambient conditions.     

Crops' responses to ozone in Mediterranean environments

The Mediterranean environment, and most of the Italian peninsula, presents some peculiarities in terms of crop response to O₃ since most physiological mechanisms activated upon O₃ exposure, such as stomatal closure, often overlap and interact with those that underlie plant adaptation to drought and hyperosmotic stress, which are typical of these environments. OTC and EDU experiments have demonstrated that O₃ causes strong yield losses when crops are grown without water limitations. However, exposure to water or saline stress significantly reduced O₃ effects on crop yield. In this review, we present the methodological approaches that have been used to study plant-ozone interactions in Italy as well as biochemical, physiological and agronomic responses for representative cropping systems of the Mediterranean climate.     

A comparison of North American and Asian exposure–response data for ozone effects on crop yields

Modelling-based studies to assess the extent and magnitude of ozone (O₃) risk to agriculture in Asia suggest that yield losses of 5–20% for important crops may be common in areas experiencing elevated O₃ concentrations. These assessments have relied on European and North American dose–response relationships and hence assumed an equivalent Asian crop response to O₃ for local cultivars, pollutant conditions and climate. To test this assumption we collated comparable dose–response data derived from fumigation, filtration and EDU experiments conducted in Asia on wheat, rice and leguminous crop species. These data are pooled and compared with equivalent North American dose–response relationships. The Asian data show that at ambient O₃ concentrations found at the study sites (which vary between ～35–75 ppb 4–8 h growing season mean), yield losses for wheat, rice and legumes range between 5–48, 3–47 and 10–65%, respectively. The results indicate that Asian grown wheat and rice cultivars are more sensitive to O₃ than the North American dose–response relationships would suggest. For legumes the scatter in the data makes it difficult to reach any equivalent conclusion in relative sensitivities. As such, existing modelling-based risk assessments may have substantially underestimated the scale of the problem in Asia through use of North American derived dose–response relationships.     

Role of ethylene diurea (EDU) in assessing impact of ozone on Vigna radiata L. plants in a suburban area of Allahabad (India)

A field study was conducted to evaluate the suitability of ethylene diurea (N-[2-(2-oxo-1-imidazolidinyl)ethyl]-N'-phenylurea; EDU) in assessing the impact of O3 on mung bean plants (Vigna radiata L. var. Malviya Jyoti) grown in suburban area of Allahabad city situated in a dry tropical region of India. EDU is a synthetic chemical having anti-ozonant property. Mean monthly O3 concentration varied between 64 and 69 microg m(-3) during the experimental period. In comparison to EDU-treated plants, non-EDU-treated plants showed significant reductions in plant growth and yield under ambient conditions. Significant favourable effects of EDU-application were observed with respect to photosynthetic pigments, soluble protein, ascorbic acid and phenol contents. EDU-treated plants maintained higher levels of pigments, protein and ascorbic acid in foliage as compared to non-EDU-treated ones. The study clearly demonstrated that EDU alleviates the unfavourable effects of O3 on mung bean plants, and therefore can be used as a tool to assess the growth and yield losses in areas having higher O3 concentrations.     

Assessing the impact of ambient ozone on growth and productivity of two cultivars of wheat in India using three rates of application of ethylenediurea (EDU)

Three rates of ethylenediurea were used to assess the impact of ambient ozone on growth and productivity of wheat (Triticum aestivum L) cultivars "Malviya 533" (M 533) and "Malviya 234" (M 234) at a suburban site near Varanasi, India, beginning in December. Wheat plants were treated with EDU at 0, 150, 300 and 450 ppm as soil drenches at 10-day intervals. EDU treatment affected plant growth, with effects varying with cultivar, age, and EDU concentration. Seed yield was improved for M 533 at 150 ppm EDU, while yield improved for M 234 at 300 and 450 ppm EDU. M 533 appears to be more resistant to ozone than M 234. Overall results confirmed that EDU is very useful in assessing the effect of ambient ozone in India.     

Cutleaf coneflower (Rudbeckia laciniata L.) response to ozone and ethylenediurea (EDU)

Cutleaf coneflower (Rudbeckia laciniata L.) seedlings were placed into open-top chambers in May, 2004 and fumigated for 12 wks. Nine chambers were fumigated with either carbon-filtered air (CF), non-filtered air (NF) or twice-ambient (2×) ozone (O₃). Ethylenediurea (EDU) was applied as a foliar spray weekly at 0 (control), 200, 400 or 600 ppm. Foliar injury occurred at ambient (30%) and elevated O₃ (100%). Elevated O₃ resulted in significant decreases in biomass and nutritive quality. Ethylenediurea reduced percent of leaves injured, but decreased root and total biomass. Foliar concentrations of cell-wall constituents were not affected by EDU alone; however, EDU × O₃ interactions were observed for total cell-wall constituents and lignocellulose fraction. Our results demonstrated that O₃ altered the physiology and productivity of cutleaf coneflower, and although reducing visible injury EDU may be phytotoxic at higher concentrations.     

Dose-response studies with the antiozonant ethylenediurea (EDU), applied as a soil drench to two growth substrates, on greenhouse-grown varieties of Phaseolus vulgaris L

To study plant growth and yield effects of the antiozonant ethylenediurea (EDU), which is frequently used for ozone crop loss assessments, dose-response studies were carried out with potted bean plants under greenhouse conditions in winter and spring. Two cultivars of Phaseolus vulgaris L., differing in sensitivity to ozone (O(3)), were grown in unfiltered air on a sandy loam rich in organic matter and on a vermiculite-clay mixture. Four treatments of EDU at concentrations from 300 to 800 mg liter(-1) were given as a soil drench during plant development. Foliar symptoms of EDU phytoxicity were observed at all doses, and plant biomass, particularly pod dry weight, was considerably reduced to increasing doses of EDU. Primary and first trifoliate leaf weight in EDU-treated plants increased as did the number of buds, indicating an extension of vegetative growth and a delay of reproductive processes. 'BBL 290' beans, which are O(3)-sensitive, were injured by EDU more than the O(3)-tolerant 'BBL 274'. The phytotoxic effects of EDU were more pronounced in the synthetic growth substrate than in field soil. In a second experiment, EDU was applied in concentrations from 100 to 400 mg liter(-1) to 'BBL 290' plants, exposed to filtered air or simulated levels of O(3) pollution. In field soil, plant growth and biomass partitioning in filtered air was only slightly altered by EDU, although leaf injury due to EDU occurred. In the vermiculite-clay mix, the biomass of most plant organs, particularly that of roots, was linearly reduced with increasing EDU doses. O(3) did not cause any alteration in plant biomass in field soil-grown and EDU-treated plants. Ozone leaf injury, which affected 67% of primary leaf area in non-treated plants, was completely suppressed by EDU doses as low as 100 mg liter(-1). This indicates that low concentrations of EDU, which do not affect plant growth in field soil, provide sufficient protection from O(3) injury. The need for careful EDU dose-response studies prior to field assessments is emphasized.     

Assessment of growth and yield losses in two Zea mays L. cultivars (quality protein maize and nonquality protein maize) under projected levels of ozone

Rapid industrialization and economic developments have increased the tropospheric ozone (O₃) budget since preindustrial times, and presently, it is supposed to be a major threat to crop productivity. Maize (Zea mays L.), a C4 plant is the third most important staple crop at global level with a great deal of economic importance. The present study was conducted to evaluate the performance of two maize cultivars [HQPM1: quality protein maize (QPM)] and [DHM117: nonquality protein maize (NQPM)] to variable O₃ doses. Experimental setup included filtered chambers, nonfiltered chambers (NFC), and two elevated doses of O₃ viz. NFC+15 ppb O₃ (NFC+15) and NFC+30 ppb O₃ (NFC+30). During initial growth period, both QPM and NQPM plants showed hormetic effect that is beneficial due to exposure of low doses of a toxicant (NFC and NFC+15 ppb O₃), but at later stages, growth attributes were negatively affected by O₃. Growth indices showed the variable pattern of photosynthate translocation under O₃ stress. Foliar injury in the form of interveinal chlorosis and reddening of leaves due to increased production of anthocyanin pigments was observed at higher concentrations of O₃. One-dimensional gel electrophoresis of leaves taken from NFC+30 showed reductions of major photosynthetic proteins, and differential response was observed between the two test cultivars. Decline in the number of male flowers at elevated O₃ doses suggested damaging effect of O₃ on reproductive structures which might be a cause of productivity losses. Variable carbon allocation pattern particularly to husk leaves, foliar injury, and damage of photosynthetic proteins led to significant reductions in economic yield at higher O₃ doses. PCA showed that both the cultivars responded more or less similarly to O₃ stress in their respective groupings of growth and yield parameters, but magnitude of their response was variable. It is further supported by difference in the significance of correlations between variables of yield and AOT40. Cultivar response reflects that QPM performed better than NQPM against elevated O₃.     

Evaluation of physiological, growth and yield responses of a tropical oil crop (Brassica campestris L. var. Kranti) under ambient ozone pollution at varying NPK levels

A field study was conducted to evaluate the impact of ambient ozone on mustard (Brassica campestris L. var. Kranti) plants grown under recommended and 1.5 times recommended NPK doses at a rural site of India using filtered (FCs) and non-filtered open top chambers (NFCs). Ambient mean O₃ concentration varied from 41.65 to 54.2 ppb during the experiment. Plants growing in FCs showed higher photosynthetic rate at both NPK levels, but higher stomatal conductance only at recommended NPK. There were improvements in growth parameters and biomass of plants in FCs as compared to NFCs at both NPK levels with higher increments at 1.5 times recommended. Seed yield and harvest index decreased significantly only at recommended NPK in NFCs. Seed quality in terms of nutrients, protein and oil contents reduced in NFCs at recommended NPK. The application of 1.5 times recommended NPK provided protection against yield loss due to ambient O₃.     

Prediction of the Vertical Profile of Ozone Based on Ground-Level Ozone Observations and Cloud Cover

Abstract

A number of statistical techniques have been used to develop models to predict high-elevation ozone (O₃) concentrations for each discrete hour of day as a function of elevation based on ground-level O₃ observations. The analyses evaluated the effect of exclusion/inclusion of cloud cover as a variable. It was found that a simple model, using the current maximum ground-level O₃ concentration and no effect of cloud cover provided a reasonable prediction of the vertical profile of O₃, based on data analyzed from O₃ sites located in North Carolina and Tennessee. The simple model provided an approach that estimates the concentration of O₃ as a function of elevation (up to 1800 m) based on the statistical results with a ±13.5 ppb prediction error, an R² of 0.56, and an index of agreement, d ₁, of 0.66. The inclusion of cloud cover resulted in a slight improvement in the model over the simple regression model. The developed models, which consist of two matrices of 24 equations (one for each hour of day for clear to partly cloudy conditions and one for cloudy conditions), can be used to estimate the vertical O₃ profile based on the inputs of the current day’s 1-hr maximum ground-level O₃ concentration and the level of cloud cover.     

Effects of Chronic Doses of Ozone on Field-Grown Loblolly Pine: Seedling Responses in the First Year

A study was designed to examine responses of loblolly pine (Pinus taeda) to chronic exposure to ozone (O₃) in the field. Seedlings of four full-sib families of loblolly pine were planted in a field near Raleigh, NC, and exposed daily (May 27 to October 24, 1985) in open-top chambers to O₃ ranging from 0.5 to 1.96 times the O₃ concentration in non-filtered (NF) air. One-fourth of the plants in each plot were removed during each of two harvests (August and October) to measure effects of O₃ on plant growth. Plants of each family exhibited foliar symptoms characteristic of O3 injury after five months of exposure to any greater-than-ambient O₃ concentration, and one family exhibited symptoms after five months of exposure to NF air. Ozone dose-plant response relationships were quantified by regression for stem height, stem diameter, biomass, and other plant morphological and yield characteristics. All relationships were linear for three families, but one family exhibited no significant growth response relationship of O₃ dose. Dose-response equations suggest a maximum growth suppression of 10 percent for NF air compared to charcoal-filtered air (i.e., 0.5 × NF) in the first season of exposure.     

An investigation of widespread ozone damage to the soybean crop in the upper Midwest determined from ground-based and satellite measurements

Elevated concentrations of ground-level ozone (O₃) are frequently measured over farmland regions in many parts of the world. While numerous experimental studies show that O₃ can significantly decrease crop productivity, independent verifications of yield losses at current ambient O₃ concentrations in rural locations are sparse. In this study, soybean crop yield data during a 5-year period over the Midwest of the United States were combined with ground and satellite O₃ measurements to provide evidence that yield losses on the order of 10% could be estimated through the use of a multiple linear regression model. Yield loss trends based on both conventional ground-based instrumentation and satellite-derived tropospheric O₃ measurements were statistically significant and were consistent with results obtained from open-top chamber experiments and an open-air experimental facility (SoyFACE, Soybean Free Air Concentration Enrichment) in central Illinois. Our analysis suggests that such losses are a relatively new phenomenon due to the increase in background tropospheric O₃ levels over recent decades. Extrapolation of these findings supports previous studies that estimate the global economic loss to the farming community of more than $10 billion annually.     

Trends and Variability of Ground-Level O3 in Connecticut over the Period 1981–1997

Abstract

The temporal and spatial characteristics of ground-level (tropospheric) O₃ measured at 10 monitoring stations in Connecticut were studied from 1981 to 1997. To detect the O₃ trend caused by changes in precursor emissions, moving average filters and a linear least-squared regression model were used to eliminate the short-term variation and effects of temperature from raw time-series O₃ data. The results showed a significant decrease in the number of days exceeding the National Ambient Air Quality Standards (NAAQS) and a small change in total O₃ concentration. The analysis indicated that the variation of daily maximum O₃, caused by changes in emissions, explained more than 10% of the total O₃ variation in Bridgeport and East Hartford during the past 17 years. Meanwhile, a strong weekly cycle was also found in O₃ time-series data, resulting in lower O₃ concentration in urban areas than in nonur-ban areas, implying that land use and land cover have impacts on the spatial distribution of ground-level O₃ in Connecticut.     

A Comparison of Nonlinear Regression and Neural Network Models for Ground-Level Ozone Forecasting

ABSTRACT

A hybrid nonlinear regression (NLR) model and a neural network (NN) model, each designed to forecast next-day maximum 1-hr average ground-level O₃ concentrations in Louisville, KY, were compared for two O₃ seasons—1998 and 1999. The model predictions were compared for the forecast mode, using forecasted meteorological data as input, and for the hindcast mode, using observed meteorological data as input. The two models performed nearly the same in the forecast mode. For the two seasons combined, the mean absolute forecast error was 12.5 ppb for the NLR model and 12.3 ppb for the NN model. The detection rate of 120 ppb threshold exceedances was 42% for each model in the forecast mode. In the hindcast mode, the NLR model performed marginally better than the NN     

An assessment of the impact of ambient ozone on field-grown crops in New Jersey using the EDU method: part 2-soybean (Glycine max (L.) Merr.)

The yields of eleven commercially grown soybean cultivars were compared in ethylenediurea (EDU)-treated and non-treated field plots in New Brunswick, New Jersey, over a 4 year period. No statistically significant difference between treatments was found for any cultivar; the inference being ambient ozone did not adversely affect soybean yield. Succeeding field experiments supported this interpretation of the data. 'Sanilac' white bean, a legume known to be more sensitive to O(3) than soybean, was found to produce a significantly greater yield in EDU-treated than non-treated plots, unlike a companion planting of 'Williams 82' soybean which did not exhibit the differential response. The results indicated that the specific EDU protocol used in the soybean experiments is capable of detecting an ozone effect in a legume. Moreover, in a concurrent greenhouse experiment the yield of EDU-treated Sanilac white bean was not significantly different from non-treated plants in the absence of ozone pollution. In a dose-response field experiment during a year of unusually high O(3) pollution, yield of 'Williams 82' increased slightly with each EDU increment up to 500 ppm and decreased at 1000 ppm. The difference between non-treated and EDU-treated plants, however, was not statistically significant. There was no evidence to suggest that the EDU concentration (500 ppm) used in previous soybean experiments reduced seed yield. Fortuitously, the tolerance of commercially-grown soybean to ambient ozone is at least partially conditioned by the practce of not irrigating the crop. The New Jersey results are in agreement with reports from Maryland, Georgia and Tennessee in which an adverse impact of ambient O(3) was not found in soybean, but contrary to a current predictive model.