Effects of nitrogen dioxide on biochemical and physiological characteristics of soybean

One month old soybean (Glycine max (L.) Merrill) cv. 'Williams' plants were exposed to nitrogen dioxide (NO2 at 0.1, 0.2, 0.3, and 0.5 microl liter(-1) and carbon filtered air (control), 7 h per day for five days, under controlled environment. Data were collected on net photosynthetic rate (PN), stomatal resistance (SR), and dark respiration rate (DR), immediately following the fifth day of exposure and 24 h after termination of exposure. Chlorophyll a (Ch a), chlorophyll b (Ch b), total chlorophyll (tot Ch) and foliar nitrogen (N) were measured before and after exposures. Growth characteristics--relative growth rate (RGR), net assimilation rate (NAR), leaf area ratio (LAR), and root shoot ratio (RSR) -- were computed for treated plants using standard growth equations. Increases of 18% and 23% in PN were observed immediately following exposure to 0.2 microl liter(-1) NO2 and after 24 h recovery period, respectively. With 0.5 microl liter(-1) NO2 treatment, reductions in PN of 23% and 50% were observed, immediately after exposure and following 24 h recovery, respectively. DR rates with 0.2 l liter(-1) treatment were higher than the control. Chlorophyll a and tot Ch showed significant reduction with 0.5 microl liter(-1) NO2 treatment. The percent reduction in Ch a and tot Ch with 0.5 microl liter(-1) NO2 were 45% and 47%, respectively. Increases in foliar nitrogen content after 0.2 and 0.3 microl liter(-1) NO2 treatments were 46% and 69%, respectively. Nitrogen dioxide at 0.5 microl liter(-1) reduced RGR and NAR by 47% and 51%, respectively. Leaf area ratio was 42% higher in 0.5 microl liter(-)1 NO2 treated plants, compared with the control; this increase was insufficient to compensate for the decrease in NAR resulting in a net decline in RGR. Nitrogen dioxide up to 0.2 microl liter(-1) increased PN and foliar-N content of soybean. With 0.5 microl liter(-1) NO2, significant decreases were observed in PN, leaf chlorophyll, foliar-N, NAR and RGR. Nitrogen dioxide up to 0.2 microl liter(-)1 has a favorable influence on overall growth characteristics of soybean; however, inhibitory effects were seen with NO2 treatment at 0.5 microl liter(-1).     

Growth and disease response of soybeans from early maturity groups to ozone and Fusarium oxysporum

The single and combined effects of ozone (O(3)) and Fusarium oxysporum on growth and disease expression of soybean genotypes differing in foliar sensitivity to O(3) were studied in the greenhouse. O(3) had no effect on root and hypocotyl rot severity of PI 153.283 (O(3)-sensitive, S) or PI 189.907 (O(3)-tolerant, T) maturity group I soybean lines. Plants of both genotypes infected with F. oxysporum and exposed to O(3) had greater reductions in relative growth rate (RGR), net assimilation rate (NAR), and had more stippled leaves per plant than Fusarium-free plants exposed to O(3). O(3) alone had a greater impact on shoot dry weight, RGR, and NAR of PI 153.283 (S) than of PI 189.907 (T). O(3) alone reduced shoot and root dry weights primarily through a depression in NAR and less through reduced leaf area. F. oxysporum alone reduced root dry weight at 35 days; however, infected plants responded with increases in root dry weight from 49 to 63 days. Similarly, F. oxysporum alone lowered early RGR but subsequent RGR decline was less rapid while NAR remained high, particularly during later sampling intervals. Infection by F. oxysporum that causes root and hypocotyl rot increased soybean sensitivity to O(3) by prolonging active vegetative growth.     

Effects of chronic ozone exposure on growth, root respiration and nutrient uptake of rice plants

To clarify the response of growth and root functions to low concentrations of ozone (O(3)), rice plants (Oryza sativa L.) were exposed to O(3) at 0.0 (control), 0.05 and 0.10 ppm for 8 weeks from vegetative to early heading stages. Exposure to 0.05 ppm O(3) tended to slightly stimulate the dry weight of whole plants up to 5 weeks and then slightly decrease the dry weight of whole plants. However, these effects were statistically significant only at 6 weeks. Exposure to 0.10 ppm O(3) reduced the dry weight of whole plants by 50% at 5 and 6 weeks, and thereafter the reduction of the dry weight of whole plants was gradually alleviated. Those changes in dry weight can be accounted for by a decrease or increase in the relative growth rate (RGR). The changes in the RGR caused by 0.05 and 0.10 ppm O(3) could be mainly attributed to the effect of O(3) on the net assimilation rate. Root/shoot ratio was lowered by both 0.05 and 0.10 ppm O(3) throughout the exposure period. The root/shoot ratio which had severely decreased at 0.10 ppm O(3) in the first half period of exposure (1-4 weeks) became close to the control in the latter part of exposure (5-8 weeks). Time-course changes in NH(4)-N root uptake rate were similar to those in the root/shoot ratio especially for 0.10 ppm O(3). On the other hand, root respiration increased from the middle to later periods. Since it is to be supposed that plants grown under stressed conditions change the ratio of plant organ weight to achieve balance between the proportion of shoots to roots in the plant and their activity for maintaining plant growth, these changes in root/shoot ratio and nitrogen uptake rate under long-term exposure can be considered to be an adaptive response to maintain rice growth under O(3) stress.     

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.     

Effects of nitrogen dioxide on leaf chlorophyll and nitrogen content of soybean

One-month-old soybean (Glycine max [L.] Merrill), cultivar 'Williams', plants were exposed to nitrogen dioxide (0.1, 0.2, 0.3 and 0.5 ppm) and carbon filtered air (control), 7 h per day, for 5 days, under a controlled environment. Leaf chlorophyll content (Ch a, Ch b, and total Ch content) and foliar nitrogen content (%N) were determined before and after the exposure. The influence of NO(2) treatments up to 0.3 ppm on leaf chlorophyll content was negligible although a stimulatory effect was evident in Ch a and total Ch content with 0.2 ppm NO(2). Marked decline in Ch content was observed with 0.5 ppm treatment; the reductions in Ch a and total Ch were 45% and 47%, respectively. Foliar-N contents of plants treated with 0.2 and 0.3 ppm NO(2) were higher than the control; plants exposed to 0.5 ppm NO(2) showed a 41% reduction in foliar-N compared to pre-exposure values.     

Differential assimilation of nitrogen dioxide by 70 taxa of roadside trees at an urban pollution level

In order to screen for the best species for mitigating nitrogen dioxide (NO2) by plants at urban levels, we investigated assimilation of nitrogen dioxide by 70 taxa of woody plants that are mostly utilized as roadside trees. They were fumigated with 15N-labeled NO2 at 0.1 microl l(-1) for 8h, and the amount of reduced nitrogen derived from NO2 (in mg Ng(-1) dry weight) in the leaves (designated NO2 assimilation capability hereafter) were determined. Data were analyzed in the comparison with the previously reported ones obtained at 4 microl l(-1) NO2. Among the 70 taxa, the value of NO2 assimilation capability differed by a factor of 122 between the highest (Prunus yedoensis; 0.061) and the lowest (Cryptomeria japonica; 0.0005). Based on the analysis of NO2 assimilation capability values at 0.1 and 4 micro l(-1) NO2, the 70 taxa of woody plants appeared to be classified into four types; those of high NO2 assimilation and high NO2 resistance, those of high NO2 assimilation but low NO2 resistance, those of low NO2 assimilation and low NO2 resistance, and those of low NO2 assimilation but high NO2 resistance. The first, second, third and fourth types include 13, 11, 35 and 11 taxa, respectively. The broad-leaf deciduous trees may have advantages of high biomass and fast growth as compared with woody plants of other habits. Thus, four broad-leaf deciduous species, Robinia pseudo-acacia, Sophora japonica, Populus nigra and Prunus lannesiana, were concluded here to be the best phytoremediators for the urban air.     

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.     

Indoor and outdoor air quality investigation at schools in Hong Kong

Five classrooms in Hong Kong (HK), air-conditioned or ceiling fans ventilated, were chosen for investigation of indoor and outdoor air quality. Parameters such as temperature, relative humidity (RH), carbon dioxide (CO2), sulphur dioxide (SO2), nitric oxide (NO), nitrogen dioxide (NO2), respirable particulate matter (PM10), formaldehyde (HCHO), and total bacteria counts were monitored indoors and outdoors simultaneously. The average respirable particulate matter concentrations were higher than the HK Objective, and the maximum indoor PM10 level exceeded 1000 microg/m3. Indoor CO2 concentrations often exceeded 1000 microl/l in air-conditioning and ceiling fan classrooms, indicating inadequate ventilation. Maximum indoor CO2 level reached 5900 microl/l during class at the classroom with cooling tower ventilation. Increasing the rate of ventilation or implementation of breaks between classes is recommended to alleviate the high CO2 level. Other pollution parameters measured in this study complied with the standards. The two most important classroom air quality problems in Hong Kong were PM10 and CO2 levels.     

Nitrogen dioxide induced changes in level of free fatty acids, triglyceride, esterified fatty acid, ganglioside and lipase activity in the guinea pig brain.

The biochemical response to controlled inhalation of nitrogen dioxide (NO2) was studied in 18 male guinea pigs. Animals were exposed to 2.5, 5.0, and 10 ppm NO2 for 2h daily for 35 consecutive days, and the results compared with six control animals exposed to filtered air for 2h daily for same period. Five biochemical parameters, including triglyceride, free fatty acids, esterified fatty acid, ganglioside and lipase activity were measured immediately after the last day of exposure. At 2.5 ppm NO2 inhalation no significant changes occurred in any region of the central nervous system (CNS). While as the dose concentration was increased to 5 and 10 ppm nitrogen dioxide, significant dose-related alteration were observed in the levels of triglyceride, free fatty acid, esterified fatty acid, ganglioside and lipase activity in the different regions of the guinea pig CNS.     

Injury and physiological responses of Larrea tridentata (DC) Coville exposed in situ to sulphur dioxide

The response of shrubs of Larrea tridentata (DEC) Coville (creosotebush) exposed to sulphur dioxide (SO(2)) was evaluated using in situ plants of the Majove Desert. Larrea was exposed to acute levels of 0.3 to 2.0 microl litre(-1) SO(2) for periods up to 13 days using field chambers or an open-air fumigation system. Plants exposed in the spring exhibited considerable leaf injury (necrosis and defoliation) when exposed to 2.0 microl litre(-1) SO(2), and in the autumn had leaf injury when exposed to >0.4microl litre(-1) SO(2). Injured plants had higher transpiration rates, less negative water pressure potentials, and/or lower photosynthetic rates than control plants. It is likely that Larrea would not be injured by the typically low SO(2) concentrations and dry environmental conditions of the Mojave Desert. However, if injury were to occur, it would be accompanied by changes in plant-water relations and photosynthesis, followed by recovery after the SO(2) stress was removed.     

Automobile Exhaust Control Devices

Irritant gases in concentrations that occur in polluted atmospheres might play a role in the degranulation and histamine release processes of mast cells in lung tissue. To test this hypothesis, young rats weighing 140-150 g were exposed to 1 ppm nitrogen dioxide for 2 hr. One group was killed immediately, and another group 24-27 hr after exposure. A third group was exposed to 0.5 ppm nitrogen dioxide for 4 hr and killed immediately. Animals serving as controls were placed for 1 hr into the exposure chamber ventilated with ambient air. Standard histological preparations were made after Carnoy’s fixative and subsequent staining with toluidine blue. The mast cells of the control animals appeared relatively intact with no evidence of disorientation. The cells of the animals exposed to NO2 and sacrificed immediately revealed rupture and loss of cytoplasmic granules with some disorientation. These changes were observed in the pleura, bronchi, and surrounding tissue with the effects more marked in the mediastinum. The mast cells of exposed animals sacrificed about 24-27 hr after discontinuing the exposure showed in some cases a combination of ruptured and intact cells with a predominance of the latter, and in other cases could not be differentiated from the controls. These findings indicate that 24 hr or more are required to reverse the acute effects of NO2 inhalation. The toxicological implications will be discussed. The release of granular substances in the lung tissue when NO2 is inhaled signifies the onset of an acute inflammation.     

Physiological responses of glyphosate-resistant and glyphosate-sensitive soybean to aminomethylphosphonic acid, a metabolite of glyphosate

Aminomethylphosphonic acid (AMPA) is formed in glyphosate-treated glyphosate-resistant (GR) and glyphosate-sensitive (GS) soybean [Glycine max (L.) Merr.] plants and is known to cause yellowing in soybean. Although, AMPA is less phytotoxic than glyphosate, its mode of action is different from that of glyphosate and is still unknown. Greenhouse studies were conducted at Stoneville, MS to determine the effects of AMPA on plant growth, chlorophyll content, photosynthesis, nodulation, nitrogenase activity, nitrate reductase activity, and shoot nitrogen content in GR and GS soybeans. AMPA was applied to one- to two-trifoliolate leaf stage soybeans at 0.1 and 1.0 kg ha(-1), representing a scenario of 10% and 100% degradation of glyphosate (1.0 kg ae ha(-1) use rate) to AMPA, respectively. Overall, AMPA effects were more pronounced at 1.0 kg ha(-1) than at 0.1 kg ha(-1) rate. Visual plant injury (18-27%) was observed on young leaves within 3d after treatment (DAT) with AMPA at the higher rate regardless of soybean type. AMPA injury peaked to 46-49% at 14 DAT and decreased to 17-18% by 28 DAT, in both soybean types. AMPA reduced the chlorophyll content by 37%, 48%, 66%, and 23% in GR soybean, and 17%, 48%, 57%, and 22% in GS soybean at 3, 7, 14, and 28 DAT, respectively. AMPA reduced the photosynthesis rate by 65%, 85%, and 77% in GR soybean and 59%, 88%, and 69% in GS soybean at 3, 7, and 14 DAT, respectively, compared to non-treated plants. Similarly, AMPA reduced stomatal conductance to water vapor and transpiration rates at 3, 7, and 14 DAT compared to non-treated plants in both soybean types. Photosynthesis rate, stomatal conductance, and transpiration rate recovered to the levels of non-treated plants by 28 DAT. Plant height and shoot dry weight at 28 DAT; nodulation, nitrogenase activity at 10 DAT, and nitrate reductase activity at 3 and 14 DAT were unaffected by AMPA. AMPA reduced root respiration and shoot nitrogen content at 10 DAT. These results suggest that a foliar application of AMPA could indirectly reduce photosynthesis through decreased chlorophyll content in GR and GS soybean up to 14 DAT, but affected plants can recover to normal growth by 28 DAT.     

Differences in ability of NO2 absorption in various broad-leaved tree species

Absorption of nitrogen dioxide (NO(2)) by various broad-leaved tree species was determined by the (15)N dilution method. The tree seedlings were continuously exposed to 0.3 ppm (microl litre(-1)) NO(2) or the mixture of 0.3 ppm NO(2) and 0.1 ppm O(3) for 30 days. The total amount of NO(2)-nitrogen absorbed by a seedling during the 30-day exposure period primarily depended on the size of the seedling. Among the tested tree species, three cultivars of Populus showed the highest rate of NO(2) absorption per unit leaf area, reaching as much as 0.3 mg N per dm(2) per day. The absorption rates for Populus cultivars were more than four times greater than those for Viburnum or Cinnamomum which had the lowest rate. A highly significant correlation was recognised between the rate of NO(2) absorption and the stomatal conductance among the species. Three cultivars of Populus which had the highest rates of NO(2) absorption were most susceptible to the mixture of NO(2) and O(3). On the contrary, Cinnamomum, Viburnum and Quercus, which showed the lowest rate of NO(2) absorption, were very tolerant to the mixed gas. These results indicate that the species difference in susceptibility to the mixture of NO(2) and O(3) was mainly determined by the difference in rate of absorption of these gases. Exposure to NO(2) alone had no detrimental effect on the tested tree species.     

Phenolic compounds content in Pinus halepensis Mill. needles: a bioindicator of air pollution

Foliar phenol concentrations (total and simple phenols) were determined in Aleppo pine (Pinus halepensis Mill.) needles collected in June 2000, from 6 sites affected by various forms of atmospheric pollutants (NO, NO(2), NO(x), O(3) and SO(2)) monitored during two months. Results show an increase in total phenol content with exposure to sulphur dioxide and a reduction with exposure to nitrogen oxide pollution. p-Coumaric acid, syringic acid and 4-hydroxybenzoic acid concentrations increase with exposure to nitrogen oxide pollution, whereas gallic acid and vanillin decrease in the presence respectively of sulphur dioxide and ozone. This in situ work confirms the major interest of using total and simple phenolic compounds of P. halepensis as biological indicators of air quality.     

Physiological and biochemical responses of two cultivars of wheat to elevated levels of CO2 and SO2, singly and in combination

In view of the present increasing trends of anthropogenic emissions of carbon dioxide (CO2) and sulphur dioxide (SO2) throughout the world, the present study was aimed at investigating the long-term influence of elevated concentrations of CO2 and SO2, singly and in combination on the physiological and biochemical characteristics of two cultivars of wheat (Triticum aestivum L. cv. Malviya 234 and HP1209). For this purpose, the plants were grown in open top chambers under field conditions and were fumigated with 600 ppm CO2, 0.06 ppm SO2 and 600 ppm CO2 + 0.06 ppm SO2 separately for 8 h daily (0800-1600 h) from germination to grain maturity. The individual treatment of SO2 advers#ely affected both the cultivars of wheat by reducing protein and starch contents. The respiration rate, total soluble sugars and total phenolics, however, increased in response to SO2. Stimulation of photosynthesis rate and reduction in stomatal conductance and transpiration rate were observed under CO2 treatment. Concentrations of total soluble sugars, starch and total phenolics increased in response to CO2 and CO2 + SO2 treatments. In combined treatment, CO2 modified the plant response to SO2 in both the cultivars. Elevated CO2 increased the photosynthesis rate under combined treatment. Higher levels of starch and soluble sugars under combined treatment provided extra carbon for SO2 detoxification. The pattern of intraspecific response of wheat to different treatments was more or less similar, but the magnitude of response differed significantly.     

Effects of ethylenediurea (EDU) on ozone-induced acceleration of foliar senescence in potato (Solanum tuberosum L.)

Experiments were conducted to examine the effects of the anti-ozonant ethylenediurea (EDU) and chronic ozone (O3) exposure on leaf physiology and senescence in an O3-sensitive potato cultivar (Solanum tuberosum L. cv. Norland). A dose-response experiment showed that an EDU concentration of 15 mg l(-1) soil (given as a soil drench) provided complete protection from accelerated foliar senescence induced by exposure to 0.1 microl l(-1) O3 for 5 h day(-1) for 11 days. EDU doses of 45 and 75 mg active ingredient l(-1) soil also gave protection but were associated with symptoms of toxicity and delayed senescence. In further experiments, plants were given 0 or 15 mg EDU l(-1) soil and exposed to clean air or 0.1 microl l(-1) O3 for 5 h day(-1) for 14 days. Chronic O3 exposure in the absence of EDU resulted in accelerated foliar senescence, characterized by early declines in net photosynthesis and Rubisco quantity in O3-treated plants relative to controls. EDU in the presence of O3 gave complete protection against symptoms of accelerated senescence. Senescence was not delayed in plants that received EDU in the absence of O3, and no symptoms of EDU toxicity were evident. The results suggest that EDU-induced tolerance to O3 was not based on 'anti-senescent' properties of this anti-ozonant.     

Sulfur dioxide effects on plants exhibiting Crassulacean Acid Metabolism

The effects of SO(2) on species exhibiting Crassulacean Acid Metabolism (CAM) were determined with short term-high concentration 'acute' greenhouse exposures (0.6 to 3.0 microl liter(-1) (ppm) SO(2) for 2 and 8 h), and long term-low concentration 'chronic' field exposures (0.35 to 0.90 microl liter(-1) SO(2) for 32 to 79 h periodically over 7 to 13 days). In the acute greenhouse exposures, visible injury was observed on Opuntia basilaris Engelm. & Bigel., exposed to 2.0 microl liter(-1) SO(2), but no injury was observed on Ananas comosus (L.) Merr., Bryophyllum blossfeldiana Poelln., Bryophyllum pinnata (Lam.) Pers., or Bryophyllum tubiflora (Harv.) Hamet, exposed to up to 2.8 microl liter(-1) SO(2) for 8 h. Stomatal conductance during the exposures averaged 0.067+/-0.021mol(-2)s(-1) for Opuntia basilaris, 0.029+/-0.008mol(-2)s(-1) for Ananas comosus, and 0.029+/-0.008mol m(-2)s(-1) for Bryophyllum pinnata. Opuntia basilaris was injured early during the day, but not at night; with the injury appearing as a white necrotic banding across just fully expanded pads. Moderately injured pads would regreen beginning 1 to 2 weeks after exposure. In chronic field exposures, no visible injury from SO(2) was observed on Opuntia basilaris, Dudleya arizonica Rose or Agave deserti Engelm. plants, grown either with supplemental irrigation or natural rainfall. In addition, in the field SO(2) had no effect on CO(2) uptake, total sulfur content, transpiration, or tissue acidity in either the light or the dark, or in irrigated vs natural rainfall plots.     

Relative sensitivity of greenhouse pot plants to long-term exposures of NO- and NO2-containing air

Thirty-five cultivars of pot plants of 20 families were exposed for 50-64 days in a greenhouse facility to either 1 microl litre(-1) NO with 0.5 microl litre(-1) NO2, or 1 microl litre(-1) NO2 with 0.1 microl litre(-1) NO for 15 h each day, with air which was free from these gases as the reference. A sensitivity ranking of the pot plants was compiled, with the highest priority on visible injuries, followed by growth reductions, primarily as a response to the NO-dominated exposures, simulating the NOx-polluted environment in direct-fired, CO2-enriched greenhouses. This treatment reduced the leaf dry weight more than the number and area of the leaves. Twenty-two cultivars were significantly injured, while two (Hibicus sp, Epipremnum pinnatum, green) were significantly improved. The NOx-sensitivity of pot plants was highest in cultivars with variegated, small or narrow leaves, and in the Moraceae family. Nine cultivars (Ficus elastica 'Robusta', F. benjamina, F. pumila 'Sonny', Dieffenbachia maculata 'Camilla', F. elastica 'Tineke', Epipremnum pinnatum 'Marble Queen', Begonia elatior 'Nelson', Cyclamen persica, Poinsettia 'Mini') were specifically sensitive to the NO-containing exposure; six were specifically sensitive to the NO2-containing exposure (F. elastica 'Robusta', Asparagus den. 'Sprengeri', Hedera helix 'Shamrock', Aspledium nidus, Aster novo-belgii, Hypoestes phyl. 'Betina'); and 12 (Soleirolia soleirolii, Asparagus den. 'Sprengeri', H. helix 'Ester', Codiaeum 'Pictum', Rosa 'Minimo Red', F. benjamina 'Starlight', Saintpaulia ionantha 'light blue', F. pumila, Rhododendron simsii, H. helix 'Shamrock', Hibiscus sp., E. pinnatum) were equally sensitive to mixtures dominated by either gas, as measured by at least one response parameter.     

The relationship between foliar injury, nitrogen metabolism, and growth parameters in ozonated soybeans

A single 12 h ozone exposure peaking at 0.20 ppm proved phytotoxic to greenhouse-grown 'Cutler 71' soybeans at each growth stage tested from V5 to R6. Visible injury occurred within 40 h on the unifoliodate leaves and middle-aged and older trifoliolates while the younger leaves were free from toxicity symptoms. In some instances visible injury was accompanied by a decrease in chlorophyll and an increase in leaf diffusive resistance. Although nitrogen fixation was not significantly altered except at early pod formation (R3), and nitrate reductase activity was significantly reduced only if the ozone exposure occurred at the time of maximal enzyme activity (V5), nitrogen content of the leaves was reduced by ozone treatment. Shoot dry weight was not affected 40 h after ozone treatment, but root dry weight was significantly reduced. Plants grown with supplemental NO(3)(-) were more sensitive to ozone than those dependent on fixed nitrogen. At plant maturity, there was no evidence of an ozone effect on shoot, root, or seed dry weight, NO(3)(-) -grown plants showed a significant increase in growth and yield over N(2)(-) plants; but no ozone effect was observed, despite the increased foliar sensitivity. Multiple ozone exposures at growth stages V3, R1 and R3 exacerbated the effects noted with a single episode and also reduced nitrogenase activity (reflected in specific and total nodule activity) and shoot and root dry weight. At plant maturity, there was again no evidence of a significant effect of multiple ozone treatment on shoot dry weight or seed yield although root weight remained low. The results would tend to support the hypothesis that older leaves of soybean do not make a significant contribution to seed yield. Although they may be injured by ozone during the reproductive phases of growth, seed yield may not be affected if the younger O(3)-tolerant leaves remain functional.     

Effects of ultraviolet-B radiation (UV-B) on growth and physiology of the dune grassland species Calamagrostis epigeios

Seedlings of Calamagrostis epigeios were exposed to four levels of UV-B radiation (280-320 nm), simulating up to 44% reduction of stratospheric ozone concentration during summertime in The Netherlands, to determine the response of this plant species to UV-B irradiation. After six weeks of UV-B treatment, total biomass of all UV-B treated plants was higher, compared to plants that had received no UV-B radiation. The increase of biomass did not appear to be the result of a stimulation of net photosynthesis. Also, transpiration rate and water use efficiency were not altered by UV-B at any exposure level. Pigment analysis of leaf extracts showed no effect of enhanced UV-B radiation on chlorophyll content and accumulation of UV absorbing pigments. UV-B irradiance, however, did reduce the transmittance of visible light (400-700 nm) of intact attached leaves, suggesting a change in anatomical characteristics of the leaves. Additionally, the importance of including an ambient UV-B treatment in indoor experiments is discussed.