Effect of long term fumigation with ozone on the turnover of the D-1 reaction center polypetide of photosystem II in spruce (Picea abies)

Long term fumigation of 4-year-old spruce trees with ozone concentrations up to 200 nl l⁻¹ has only minor effects on the photosynthetic activities measured as chlorophyll a fluorescence. Nevertheless, it drastically changes the turnover of the D-1 reaction center polypeptide of photosystem II. During summer, fumigation with ozone for 2 weeks resulted in an almost 4-fold stimulation of the light dependent incorporation of [¹⁴C] leucine into the D-1 protein in the exposed trees. The amount of immunodetectable D-1 protein remained constant when based on chlorophyll. This indicates that exposure to ozone stimulates both the synthesis and the degradation of the D-1 protein. When spruce trees were exposed during winter for 4 weeks to 100 and 200 nl l⁻¹ ozone, respectively, an almost 3-fold increase of the amount of immunodetectable D-1 protein per chlorophyll in the exposed trees was observed. This can be explained by a varying stimulation of D-1 protein synthesis and degradation depending on the different physiological conditions. Since so far the D-1 protein has been found only as a component of photosystem II reaction centers, one has to assume that the relative content of photosystem II reaction centers also increases under certain stress conditions. The increased turnover of the D-1 protein in trees exposed to ozone explains the synergistic effects of stress conditions and high light intensities often observed in the field.     

Responses of Photosynthesis,Carbohydrates and Antioxidants in Needles of Norway Spruce to Slow and Rapid Changes in Ozone

The goal of the present study was to examine the effects of slow and rapid changes of ozone (O₃) concentrations on the physiological behaviour of current-year needles of Norway spruce (Picea abies (L.) Karst.). For this purpose five-year-old spruce seedlings were exposed in growth chambers for 49 days to either charcoal-filtered air, slowly increasing O₃ concentrations from zero up to 100 nl I^?1 in weekly steps of 25 nl I^?1, or immediately to 100 nl I^?1 of O₃. During the investigation period gas exchange, carbohydrate and antioxidant contents of the current flush were measured. In needles which experienced slowly increasing O₃ concentrations, cumulative O₃ uptake was approximately 30 % lower than in needles continuously fumigated with 100 nl I^?1 of O₃. The higher 0₃ uptake in the permanent 100 nl I^?1 O₃ treatment caused a pronounced decline in net photosynthesis, in the efficiency of CO₂ uptake and in the starch content of the seedlings. Initially the ascorbate pool increased, but after 5 weeks of exposure ascorbate concentrations declined and were comparable to values obtained in charcoal-filtered controls, while the thiol contents were enhanced during fumigation with permanent 100 nl I-^?1 O₃. On the contrary, slowly increasing O₃ caused a significant increase in total needle ascorbate throughout the fumigation period, which probably prevented an O₃-induced decline in the photosynthetic machinery as photosynthesis was not affected although the thiol contents were not enhanced. Furthermore, starch content was slightly higher than in O₃-free controls. These results suggest that seedlings of Norway spruce have the possibility to acclimate to O₃ stress, as slowly increasing O₃ concentrations seemed to increase resistance and the seedlings were able to compensate.     

Concentration dependencies of some effects of ethylene on etiolated pea, peanut, bean, and cotton seedlings

The effects of a series of concentrations of ethylene (10, 20, 40, to 10,240 nl/l) on elongation, diameter, and geotropism of the stems and roots of etiolated seedlings of Pisum sativum L., Arachis hypogea L., Phaseolus vulgaris L., and Gossypium hirsutum L. were measured or observed. Of the 24 possible responses, 4 were unaffected at the concentrations used, 5 were affected slightly, and the remaining responses exhibited a 14-fold range of apparent half-maximum concentration dependencies (i.e. 95 nl/l for the effect on pea epicotyl geotropism to 1350 nl/l for the promotion of cotton hypocotyl diameter). Six or possibly eight of these responses appear to have the same concentration dependencies while the others fell in pairs or as individual responses. The data, if interpreted in a manner analogous to enzyme kinetics, are indicative of more than one primary mechanism for ethylene action in plants.     

The Sensitivity of Net Photosynthesis in Several Plant Species to Short-term Fumigation with Sulphur Dioxide

The effects of exposure of up to 2 h with sulphur dioxide ona range of plant species was observed by measuring changes inthe rate of net photosynthesis under closely controlled environmentalconditions. Ryegrass, Lolium perenne ‘S23’ was thespecies most sensitive to SO₂; significant inhibition was detectedat 200 nl l^–1. Fumigations at 300 nl l^–1 also inhibitedphotosynthesis in field bean (Vicia faba cv. ‘Three FoldWhite’ and ‘Blaze’) and in barley (Hordeumvulgare cv. ‘Sonja’). No effect was detected inwheat (Triticum aestivum cv. ‘Virtue’) at concentrationsup to 600 nl l^–1 SO₂, or in oil-seed rape (Brassica napuscv. ‘Rafal’) except at 800 nl l^–1 SO₂). Recoverycommenced immediately after the fumigation was terminated andwas complete within 2 h when inhibition had not exceeded 20%during the SO₂ treatment. Key words: Sulphur dioxide, short-term fumigation, photosynthesis     

Ethylene effects on growing and gravireacting maize root segments

The effects of ethylene pretreatments (500 nl/l for 1 h) and treatments (100 nl/1 to 1000 nl/l for 6 h) on elongation and gravireaction of apical maize root segments were tested in light and in the dark. Ethylene was found to affect weakly root elongation and gravireaction, but to induce strong curvatures for root growing vertically.     

Filtering impacts of larval and sessile zebra mussels (Dreissena polymorpha) in western Lake Erie

Summary We assessed the feeding biology of veliger larvae of the introduced zebra mussel (Dreissena polymorpha Pallas) in laboratory experiments using inert microspheres as food analogues. Mean clearance rate on 2.87-m beads ranged between 247 and 420 L veliger^–1 day^–1. Clearance rate was unrelated to bead concentration up to 100 beads L^–1, but was positively correlated with veliger shell length. Clearance rates of Dreissena veligers are within the range of those reported for marine bivalve veligers of similar size and for herbivorous Great Lakes microzooplankton, but are orders of magnitude lower than those of settled, conspecific adults. The impact of settled zebra mussel grazing activities on phytoplankton stocks may be up to 1162 times greater than that exerted by veliger populations in western Lake Erie. Based on 1990 size-frequency distributions and associated literature-derived clearance rates, reef-associated Dreissena populations in western Lake Erie (mean depth 7 m) possess a tremendous potential to filter the water column (up to 132 m³ m^–2 day^–1) and redirect energy from pelagic to benthic foodwebs. Preliminary analyses indicate that chlorophyll a concentration is strongly depleted (<1 g L^–1) above Dreissena beds in western Lake Erie.     

Relationship of antioxidant enzymes to ozone tolerance in branches of mature ponderosa pine (Pinus ponderosa) trees exposed to long-term, low concentration, ozone fumigation and acid precipitation

Seasonal activity of superoxide dismutase (SOD, EC 1.15.1.1). ascorbate peroxidase (APOD, EC 1.11.1.11) and guaiacol-oxidizing enzymes (GPODs, EC 1.11.1.7) was examined in needles of 12- to 15-year-old ponderosa pine (Pinus ponderosa Laws.) trees which received ozone (O₃) and acid precipitation treatment. Individual branches were enclosed in branch exposure chambers delivering either charcoal-filtered (O₃-reduced) air, ambient air, or air with twice ambient (2 x ambient) concentrations of O₃. Acid precipitation treatments were rain of pH 3.0 or 5.1 or no rain. Changes in antioxidant enzyme activity were not a consistent response to O₃ fumigation or acid precipitation, but when observed, they occurred most often in the O₃-sensitive clone and in symptomatic, fumigated branches. In the second year of fumigation. O₃ fleck symptoms appeared on needles of the sensitive clone as early as July and APOD activities were significantly increased by O₃ at all sampling dates. In the tolerant clone, antioxidant enzyme activities were not significantly changed by O₃ in the first season of fumigation (March to December 1990), not even during an episode when ambient O₃ concentrations reached 125 nl 1^?1 (240 nl 1^?1 in 2x ambient chambers). No foliar symptoms were observed on needles of the tolerant clone during this year. However, in the second year of fumigation (1992), O₃ fleck symptoms were observed on the tolerant clone and APOD activities were significantly increased in previous-year needles. The tolerant clone had SOD, APOD, and GPOD activities at least 40% higher than those of the sensitive clone before fumigation and 65, 178, and 119% higher, respectively, during both years of fumigation. The higher constitutive levels of these enzymes may have protected against foliar injury in 1990, however in 1992 we concluded that the stimulations in antioxidant enzyme activities observed in symptomatic branches of both clones were a consequence of O₃ injury. Total (intra- and extracellular) activities of the antioxidant enzymes did not appear to be good indicators of O₃ tolerance. Phenotypically, the O₃-tolerant clone was much more vigorous and in both years of fumigation, gas exchange rates were 30 to 71% higher than in the sensitive clone (P. D. Anderson, unpublished data). The greater vigor of the tolerant clone may allow more carbon allocation to protective and repair processes which include, but are not restricted to, the turnover of antioxidant enzymes and metabolites.     

The influence of ionic concentration on population parameters,development time,activity, and respiration rate of Arctodiaptomus spinosus (Daday) (Calanoida,Copepoda)

Summary For A. spinosus, significant correlations were found between population parameters and the ecological parameters of alkalinity and temperature. For the birth rate, 43.33% of variance is explained by alkalinity and 56.33% by alkalinity and temperature. Alkalinity, and alkalinity and temperature combined account for 48.22% and 70.23% respectively of the observed variances in nauplii hatching. It was demonstrated that, provided sufficient time for adaptation is allowed, the activity of individuals is not affected by differences in the alkalinity of the medium within the normal range of concentration. Embryonic development does not appear to be influenced by the ionic concentration, but significant increases in time of development of the nauplii (at alkalinity 100 meq/l: 8.8 days/20°C) and of the copepodites (at alkalinity 100 meq/l: 12.6 days/20°C) were recorded at both low and high ionic concentrations. The lowest respiration rates (3.7 nl O₂·h^-1·g dw^-1/20°C) occurred at an alkalinity of 100 meq/l. Rates increased at lower (10 meq/l: 5.7 nl) and at higher (400 meq/l: 11.9 nl) alkalinities.     

Stimulation of the diadinoxanthin cycle by UV-B radiation in the diatom Phaeodactylum tricornutum

In this study we show that the diadinoxanthin cycle in the diatom Phaeodactylum tricornutum is stimulated by mild UV-B radiation. High steady state concentrations of diatoxanthin established during a period of strong actinic illumination with white light (300 mol photons m-2 s-1 PAR) are further increased if weak UV-B (3 mol photons m-2 s-1) is additionally applied. Short term increases in the diatoxanthin concentration caused by UV-B strongly correlate with a stoichiometric decrease in diadinoxanthin. The UV-B dependent increase in diatoxanthin is correlated with a concommitant enhancement of non-photochemical quenching of chlorophyll fluorescence and a decrease in the quantum efficiency of oxygen evolution. This indicates that UV-B induced diatoxanthin functions in thermal energy dissipation. Possible scenarios for a stimulation of the diadinoxanthin cycle by UV-B are discussed.     

Responses of banana fruit to treatment with 1-methylcyclopropene

Experiments were conducted to determine levels of 1-methylcyclopropene (1-MCP) exposure needed to prevent ethylene-stimulated banana fruit ripening, characterise responses of ethylene-treated fruit to subsequent treatment with 1-MCP, and to test effects of subsequent ethylene treatment on 1-MCP-treated fruit softening. Fruit softening was measured at 20°C and 90% relative humidity. One hour exposure at 20°C to 1000 nl 1-MCP/l essentially eliminated ethylene-stimulated ripening effects. Exposure for 12 h at 20°C to just 50 nl 1-MCP/l was similarly effective. Fruit ripening initiated by ethylene treatment could also be delayed with subsequent 1-MCP treatment. However, 1-MCP treatment only slowed down ripening of ethylene-treated fruit when applied at 1 day after ethylene and was ineffective when applied 3 or 5 days after ethylene treatment. The ripening response of fruit treated with 1-MCP and subsequently treated with ethylene varied with interval time between 1-MCP and ethylene treatments. As time increased, the response of 1-MCP-treated fruit to ethylene was enhanced. Responses to 0.1, 1, 10 or 100 µl ethylene/l concentrations were similar. Enzyme kinetic analysis applied to 1-MCP effects on ethylene-induced softening of banana fruit suggested that 1-MCP inhibition is by noncompetitive antagonism of ethylene binding.     

The response of sensitive and tolerant clones of Populus tremuloides to dynamic ozone exposure under controlled environmental conditions

Both sensitive and tolerant clones of aspen ( Populus tremuloides ) were exposed to ozone using four different exposure regimes under controlled environmental conditions. Based on data on ambient ozone from 10 cities in the USA, three treatments of 4-wk exposure to the same SUM06 (an accumulation of hourly O₃ concentrations greater than 0.06 ml l⁻¹) were constructed. The regimes allowed us to investigate: (a) the importance of long (3 wk, treatment 1) versus short (1 wk, treatment 2) duration of regimes with high peaks; (b) the effect of treatments with variable peak occurrence (treatments 1 and 2) versus uniform peak occurrence (treatment 3) during the exposure period. Nonfumigated control plants were maintained at ozone concentrations <10 nl l⁻¹. Bifacial black necrosis, a typical symptom of ozone injury on aspen leaves, occurred on both clones after 2 wk exposure. Up to 60% of the leaves on the sensitive clone were injured, with an average of 6% of total leaf area injured. In the tolerant clone only 10% of the leaves were injured, with less than 1% of the total leaf area symptomatic. The severity of injury was consistently greatest in treatment 2, followed by treatments 1 and 3, respectively. The interval between peak exposures was less important than the occurrence of peaks versus a stable maximum concentration. Premature leaf abscission occurred in the sensitive clone. Measures of gas exchange demonstrated reduced photosynthesis under ozone fumigation, but exposure regime was not a significant factor. Concentrations of two antioxidants, ascorbic acid and glutathione, were almost always greater in the resistant than in the sensitive clone, but the differences were not statistically significant. The levels of these antioxidants in aspen leaves did not change with ozone fumigation or leaf age.     

Probing the role of endogenous ethylene in the degreening of citrus fruit with ethylene antagonists

The ethylene antagonists, 2,5-norbornadiene (NBD) and silver nitrate, were used to probe the involvement of endogenous ethylene in the natural degreening of citrus fruit. Mature-green, detached Shamouti orange (Citrus sinensis L. Osbeck) fruit were treated with NBD vapor or dipped in solutions of silver nitrate. More than 80% of the chlorophyll was lost from control fruit after 8 days. NBD (0.11 mmole/liter) inhibited the loss of chlorophyll by 60%. NBD also antagonized the degreening induced by exogenous ethylene by 50%. Silver nitrate (0.1 mM) inhibited the loss of chlorophyll by 55%. Ethylene evolution of mature, green detached fruit was <2 nl.fruit^-1.h^-1 (ca. 13.5 nl.Kg^-1FW.h^-1) and did not change significantly for 7 days after harvest. NBD concentrations up to 0.22 mmole/liter did not enhance ethylene evolution. Not with-standing the extremely low amounts of ethylene evolved, the inhibition of degreening by NBD and silver nitrate suggests that endogenous ethylene is involved in the control of this process in mature citrus fruit.     

Potential of Allyl Isothiocyanate to Control Rhizoctonia solani Seedling Damping Off and Seedling Blight in Transplant Production

The synthetic mustered flavouring essential oil, allyl isothiocyanate (AITC), was evaluated for its effect on suppression of Rhizoctonia solani growth in vitro, and in field soils for reducing inoculum density, saprophytic substrate colonization and seedling damping off and blight using snap bean and cabbage as indicator plants. In vitro growth was completely inhibited at the concentration of 50 μl/l. Inoculum density and saprophytic substrate colonization by the fungus in soil were not affected by AITC concentrations of 50 or 75 μl/kg soil. The inoculum density estimation by the use of soil‐drop technique created an artefact leading to an erroneous conclusion that the fungus was eradicated from soil within 1–3 days after AITC treatment at 150 or 200 μl/kg soil. The saprophytic substrate colonization showed that although the activity of R. solani was greatly reduced, the fungus still colonized 45% of the substrate units at these concentrations, and up to 100% at lower concentrations within 1 day after treatment. At higher concentrations the recovery rate from the substrates gradually declined over time to <6%. Drenching R. solani infested sandy‐loam or silty‐clay‐loam soil with water containing the emulsified AITC to provide 150 or 200 μl/l soil, a few days prior to planting, gave over 90% disease control in snap bean and cabbage, with no apparent phytotoxic effect. The effect of AITC was not influenced by the physical soil texture. AITC appears to have a good potential to replace methyl bromide fumigation of the substrate used for transplant production.     

Acetic Acid, Ethanol and Steam Effects on the Growth of Botrytis cinerea in vitro and Combination of Steam and Modified Atmosphere Packaging to Control Decay in Kiwifruit

The effects of acetic acid fumigation, ethanol fumigation, and steam heat treatment on growth of Botrytis cinerea in vitro were investigated. The effect of steam heat treatments in combination with modified atmosphere packaging (MAP) on Botrytis decay development on 'Hayward' kiwifruit was also studied. The fungus was grown in Petri dishes on potato dextrose agar. Ethanol fumigation with 100  μ l/l for 3 or 6 min, or 200  μ l/l for 6 min enhanced the growth of B. cinerea . The effect of acetic acid on growth of B. cinerea was time and dosage-dependent. Fumigation with 1  μ l/l for 6 min, 2  μ l/l for 3 min, and 4  μ l/l for 3 min promoted radial growth of the fungus when compared to the growth of the untreated control. Fumigation with 2  μ l/l for 6 min delayed the growth of the fungus for the first 6 days, while fumigation with 6  μ l/l for 3 min delayed the growth of the fungus after the sixth day. Fumigation with 4 or 6  μ l/l acetic acid for 6 min, and 8  μ l/l acetic acid for 3 or 6 min resulted in complete inhibition of fungal growth. Steam heat treatment at 45°C for 6 min, and at 48, 51, and 54°C for 3 or 6 min completely inhibited fungal growth in vitro . Furthermore, steam treatments at 47, 50, and 53°C for 3 or 6 min completely inhibited decay at the stem end of kiwifruit kept at 10°C in MAP for 12 days. However, none of the steam treatments inhibited decay in wounds on the surface of the fruit kept in MAP.     

Responses of citrus plants to ozone: leaf biochemistry, antioxidant mechanisms and lipid peroxidation.

The effects of ozone upon 3-year-old trees of Clementina mandarin (Citrus clementina Hort. ex Tan.) cv. Marisol exposed for 12 months to ambient (10 nl l(-1)) and high (30 and 65 nl l(-1)) concentrations in open top chambers (OTCs) were investigated. The data showed that in leaves, ozone reduced total chlorophylls, carotenoid and carbohydrate concentration, and increased 1-aminocyclopropane-1-carboxylic acid (ACC) content and ethylene production. In treated plants, the ascorbate leaf pool was decreased, while lipid peroxidation and solute leakage were significantly higher than in ozone-free controls. The data indicated that ozone triggered protective mechanisms against oxidative stress in citrus.     

Bacterivory by starved marine heterotrophic nanoflagellates of two species which feed differently, estimated by uptake of dual radioactive-labelled bacteria

Abstract: The rates of ingestion of bacteria and of accumulation of bacterial biomass by hungry Pteridomonas danica and Paraphysomonas imperforata were measured using dual radioactive-labelled bacteria in experiments lasting 4–8 h. Pteridomonas continuously consumed 4–5 bacteria h⁻¹ throughout experiments lasting 8 h, irrespective of bacterial concentration above a threshold of about 5 × 10⁵ bacteria ml⁻¹, and continued to catch bacteria even below this density. The clearance rate of about 1 nl cell⁻¹ h⁻¹ at higher bacterial concentrations increased three or four times as bacterial numbers fell. Paraphysomonas cells, with only half the biomass of Pteridomonas , ingested up to 10 bacteria h⁻¹ at high bacterial concentrations, and gradually reduced the feeding rate, effectively ceasing to feed at 10⁶ bacteria ml⁻¹; their initial clearance rate of 1–2.5 nl cell⁻¹ h⁻¹ subsequently fell as low as 0.1 nl cell⁻¹ h⁻¹. Estimation of feeding rate by extrapolation from short-term experiments on such flagellates requires extreme caution. These flagellates, starved to levels typical of the natural environment, accumulated ingested bacterial biomass at an efficiency of between 16 and 21%, indicating that in nature they would recycle 80% or more of the nutrients contained in their food.     

Effects of air pollutants on the photosynthetic capacity of young Norway spruce trees. Response of single needle age classes during and after different treatments with O3, SO2, or NO2

Summary Potted young Norway spruce trees were exposed to different concentrations of the air pollutants ozone, sulphur dioxide and nitrogen dioxide under completely controlled environmental conditions. After the treatment, the potted trees were kept outdoors. Measurements of the maximum photosynthetic capacity (A₂₅₀₀) were performed with current-year and 1-year-old needles during and after exposure of the trees. In trees fumigated with nitrogen oxides no damage was found at the concentrations used, and the trees' ability to fix carbon dioxide was increased. Using SO₂, a rapid and marked decrease in A₂₅₀₀ was obtained within the first days of the experiment. This decrease did not continue further, but was reversed upon cessation of the fumigation. However, a clear dose-dependent decrease in A₂₅₀₀ occurred when trees were fumigated continuously with an ozone concentration of 450 nl l^–1 or more. The effect of ozone was not reversible, but continued during post-culture of the trees.     

Cellulase Activity, Endogenous Abscisic Acid, and Ethylene in Four Citrus Cultivars during Maturation

At maturity, the fruit of two early maturing orange cultivars, Hamlin and Pineapple (Citrus sinensis [L.] Osbeck), contained more ethylene and abscisic acid than the late maturing Valencia and Lamb Summer (C. sinensis [L.] Osbeck) cultivars. Ethylene (up to 95 nl/l in internal atmosphere) and abscisic (50 μg/kg dry weight flavedo) increased most rapidly in Pineapple, leading to increased cellulase activity and loosening of the fruit. Fruit of the two late maturing cultivars contained less than 25 nl/1 ethylene and 40 μg abscisic acid/kg dry weight of flavedo at peak maturity. Cellulase activity and loosening of the fruit of these late maturing cultivars was slight.     

Acaricidal activity and repellency of essential oil from Piper aduncum and its components against Tetranychus urticae

The chemical composition of essential oil of leaves of Piper aduncum L., growing wild in a fragment of the Atlantic Rainforest biome in northeastern Brazil, was determined through gas chromatography-mass spectrometry. The acaricidal activity and repellency of the essential oil and its components [dillapiole (0.28 g/ml), α-humulene (0.016 g/ml), (E)-nerolidol (0.0007 g/ml) and β-caryophyllene (0.0021 g/ml)] were evaluated in the laboratory against adults of Tetranychus urticae Koch. The mites were more susceptible to the oil in fumigation tests (LC(50) = 0.01 μl/l of air) than in contact test with closed Petri dish (LC(50) = 7.17 μl/ml); mortality was reduced by approximately 50 % in the latter test. The repellent action of the oil and toxicity by fumigation and contact did not differ significantly from the positive control (eugenol). The repellent activity was attributed to the components (E)-nerolidol, α-humulene and β-caryophyllene, whereas toxicity by fumigation and contact was attributed to β-caryophyllene. The effect of Piper oil and the role of its components regarding host plant preference with a two-choice leaf disk test are also discussed.