Differences in growth, leaf senescence and injury, and stomatal density in birch (Betula pendula Roth.) in relation to ambient levels of ozone in Finland

The differences in growth, leaf senescence, visible ozone injuries and stomatal density between one coastal site (natural ozone) and two inland sites (natural and elevated ozone) in Finland were determined for saplings of Betula pendula clones grown under open-field conditions during two growing seasons. Responses in growth, leaf senescence, visible injuries, and stomatal density were determined in relation to cumulative ozone exposure accumulated over the thresholds of 30, 40 and 50 ppb (10(9)) during the exposure period. In addition, the effects of the different ozone exposures on ultrastructure of chloroplasts were studied. Increasing ozone exposure resulted in reduced shoot dry weight, stimulated (first year) or reduced (second year) height growth, accelerated autumn yellowing of leaves, increased stomatal density, visible symptoms and chloroplast injuries, and increased number and size of plastoglobuli. Newly expanded mature leaves in midsummer were more sensitive to ozone episodes than younger developing leaves in the early growing season. In most parameters, the best correlation was achieved with the exposure index AOT30. Ozone risk for birch is highest in the southern coastal area of Finland, where background ozone concentrations are higher than in inland sites.     

Tibouchina pulchra (Cham.) Cogn., a native Atlantic Forest species, as a bio-indicator of ozone: visible injury

Tibouchina pulchra saplings were exposed to carbon filtered air (CF), ambient non-filtered air (NF) and ambient non-filtered air+40 ppb ozone (NF+O3) 8 h per day during two months. The AOT40 values at the end of the experiment were 48, 910 and 12,895 ppb h(-1), respectively, for the three treatments. After 25 days of exposure (AOT40=3871 ppb h(-1)), interveinal red stippling appeared in plants in the NF+O3 chamber. In the NF chamber, symptoms were observed only after 60 days of exposure (AOT40=910 ppb h(-1)). After 60 days, injured leaves per plant corresponded to 19% in NF+O3 and 1% in the NF treatment; and the average leaf area injured was 7% within the NF+O3 and 0.2% within the NF treatment. The extent of leaf area injured (leaf injury index) was mostly explained by the accumulated exposure of ozone (r2=0.89; p<0.05).     

Psidium guajava 'Paluma' (the guava plant) as a new bio-indicator of ozone in the tropics

Psidium guajava 'Paluma' saplings were exposed to carbon filtered air (CF), ambient non-filtered air (NF), and ambient non-filtered air+40ppb ozone (NF+O(3)) 8h per day during two months. The AOT40 values at the end of the experiment were 48, 910 and 12 895ppbh(-1), respectively for the three treatments. After 5 days of exposure (AOT40=1497ppbh(-1)), interveinal red stippling appeared in plants in the NF+O(3) chamber. In the NF chamber, symptoms were observed only after 40 days of exposure (AOT40=880ppbh(-1)). After 60 days, injured leaves per plant corresponded to 86% in NF+O(3) and 25% in the NF treatment, and the average leaf area injured was 45% in NF+O(3) and 5% in the NF treatment. The extent of leaf area injured (leaf injury index) was explained mainly by the accumulated exposure of ozone (r(2)=0.91; p<0.05).     

Detached leaf culture: viability to evaluate 2,4-D toxicity symptoms in cotton apex leaves

To study the viability of detached leaf culture technique, studies were carried out with detached leaves from cotton apex (true trilobed leaves). The prepared leaves were sprayed with 2,4-D amine and ester, at rates of 10, 30, 70, and 100% of the recommended doses. Detached leaves without herbicide spray were used as controls. Simultaneously, a greenhouse experiment was conducted with the same treatments as used for the detached leaves experiment. Toxicity was measured through a 0-to-5 grading according to the percentage of affected leaf area in the detached leaves experiment or examining the affected rate of whole plant as indicated in the greenhouse. Results showed that the ester form of the herbicide induced earlier and more severe toxicity symptoms in detached leaves and greenhouse grown plants. Positive and significant correlations (p < 0.001) were found between toxicity results obtained at 7 and 14 days after application in detached leaves and greenhouse plants (r = 0.97 and 0.92, respectively). Negative, significant correlations (p < 0.005) were found between the toxicity levels found at 7 and 14 days after application in detached leaves and dry matter of cotton plants grown in the greenhouse (r= - 0.92 and -0.92, respectively).     

Alteration in uptake and translocation of essential nutrients in cabbage by excess lead

To observe the tolerance limit of lead phytotoxicity in cabbage (Brassica oleracea L.) var. Golden Aker plants were grown in refined sand with complete nutrient solution for 41 days. On 42nd day, pots with plants were separated into six lots. One lot was allowed to grow as such and was treated as control, in rest of the five lots, lead was applied at 0.1, 0.2, 0.4, 0.5 and 1.0 mM as lead nitrate. At d 75 (34 days after metal exposure), the lead toxicity symptoms as restricted growth was observed on plants at 1.0 mM lead supply. Excess lead (0.5 and 1.0 mM) developed interveinal chlorosis along the margins of young leaves. The affected leaves were reduced in size giving plant a rosette like appearance. Head size was markedly reduced at these (0.5 and 1.0 mM) levels of lead. At 0.5 mM the intensity of symptoms was markedly low. With an increase in lead supply, the concentration of lead and zinc was increased whereas that of P, S, Fe, Mn and Cu were decreased in various parts of cabbage. At 1.0 mM Pb, the concentration of lead was highest in roots and lowest in head. In leaves of cabbage the threshold of toxicity and toxicity values were 150 and 320 microg g-1 dry matter, respectively.     

Impact of ozone on the growth of birch (Betula pendula) saplings

Saplings of one half-sib family of birch, Betula pendula, were exposed to three levels of ozone in open-top chambers (OTCs) during two growing seasons 1997-1998. The ozone treatments were non-filtered air (NF, accumulated daylight AOT40 over the two growing seasons of 3.0 l l-1 h), non-filtered air with extra ozone (NF+, accumulated daylight AOT40 of 27.3 l l-1 h) and non-filtered air with additional extra ozone (NF++, accumulated daylight AOT40 of 120 l l-1 h). The birch saplings, including the roots, were harvested after the first and second growing seasons. After the first growing season, the NF++ treatment reduced the total wood biomass by 22%, relative to the NF treatment. There was no further reduction of the total wood biomass in the NF++ treatment after the second growing season. The root biomass was reduced by 30% after the first growing season. The shoot/root ratio, as well as the proportional biomass of leaves, were increased by ozone during both years. The ozone impact on the relative growth rate was estimated to -2% per 10 l l-1 h daylight AOT40 per growing season.     

Growth stimulation of Triticum aestivum seedlings under Cr-stresses by non-rhizospheric pseudomonad strains

Four chromium-resistant non-rhizospheric strains SPCr-1, SPCr-2, SPCr-3 and SPCr-4 (Pseudomonads), which were isolated from the effluents of an ICI paint factory and could tolerate 2-3 mg ml(-1) chromium in a minimal medium and 40 mg ml(-1) in a rich medium, were used to inoculate seeds of Triticum aestivum. Both inoculated and non-inoculated seeds were germinated and grown under different concentrations of chromium salts (K2CrO4, 0, 100, 250, 500; CrCl3, 0, 250, 500, 1000 microg ml(-1)). Germination and growth parameters were severely affected by chromium-salts. K2CrO4 had more drastic effects than CrCl3 treatments. Seedlings had a hard and brittle texture and showed symptoms of hypertrophy. Brown spots on leaves and stems were visible and the tips of leaves were bifurcated and curled. The root system was also impaired, ranging from a browning of the tip to complete destruction of cortical tissues. Under chromium-stress conditions, inoculated plants had significantly better germination and growth as compared to non-inoculated treatments. Bacterial growth enhancement of seedlings was associated with reduced chromium-uptake, increased auxin content and the formation of stress specific proteins. With bacterial inoculations, symptoms of chromium toxicity were reversed, especially at lower concentrations of chromium salts.     

Phytotoxicity to and uptake of flumequine used in intensive aquaculture on the aquatic weed, Lythrum salicaria L

Phytotoxicity of Flumequine on the aquatic weed Lythrum salicaria L. was determined by two laboratory models: a single concentration test, by which the effects of 100 mg l-1 were evaluated after 10, 20, 30 days and a multiple concentration test, by which the effects of 5000-1000-500-100-50 micrograms l-1 were evaluated after 35-day exposure. 100 mg l-1 are highly toxic and significantly decrease the growth of plants; this effect increases with time. Concentrations between 5000 and 50 micrograms l-1 induced hormesis in plants, by significantly increasing mean number and dimension of leaves and secondary roots. The effect is the highest at 50 micrograms l-1 and decreases with increase in concentration. Both toxic effect and hormesis can be related to plant drug uptake, quite high, in the order of micrograms g-1. The ecological implication of Flumequine contamination in aquatic environments and the possible use of Lythrum salicaria for bioremediation and/or monitoring technique are discussed.     

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

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

Detached Leaf Culture

Abstract

To study the viability of detached leaf culture technique, studies were carried out with detached leaves from cotton apex (true trilobed leaves). The prepared leaves were sprayed with 2,4-D amine and ester, at rates of 10, 30, 70, and 100% of the recommended doses. Detached leaves without herbicide spray were used as controls. Simultaneously, a greenhouse experiment was conducted with the same treatments as used for the detached leaves experiment. Toxicity was measured through a 0-to-5 grading according to the percentage of affected leaf area in the detached leaves experiment or examining the affected rate of whole plant as indicated in the greenhouse. Results showed that the ester form of the herbicide induced earlier and more severe toxicity symptoms in detached leaves and greenhouse grown plants. Positive and significant correlations (p < 0.001) were found between toxicity results obtained at 7 and 14 days after application in detached leaves and greenhouse plants (r = 0.97 and 0.92, respectively). Negative, significant correlations (p < 0.005) were found between the toxicity levels found at 7 and 14 days after application in detached leaves and dry matter of cotton plants grown in the greenhouse (r = ?0.92 and ?0.92, respectively).     

Initial risk assessment for three beta-blockers found in the aquatic environment

Daphnia magna, Desmodesmus subspicatus and Lemna minor were used to determine the ecotoxicity of beta-blockers. Propranolol was the most toxic substance, with EC50s of 7.7 mg l-1 in the Daphnia test and 0.73 mg l-1 in the algal test. The toxicity of atenolol and metoprolol in the Daphnia test was very low, with EC50s of 313 and 438 mg l-1, respectively. In the algal test, metoprolol (EC50: 7.9 mg l-1) was more toxic than atenolol (EC50: 620 mg l-1). Lemna minor was the least sensitive species. All substances showed PEC/PNEC ratios <1, whereas propranolol with a ratio of 0.81 seems to be the most relevant substance and I recommend to perform chronic biotests to refine the estimate of the environmental risk. Taking bioconcentration factors (BCF) into account, resulting internal effect concentrations (IEC) differ only slightly, which means that the differences in the EC50 depend mainly on the diverse logP levels causing narcosis via disruption of membrane integrity. Predictions of toxicity using ECOSAR were close to the empirically measured data. Mixture toxicity of the three compounds could be predicted accurately using the concept of concentration addition.     

Bioaccumulation and physiological effects of mercury in Sesbania drummondii

The accumulation of mercury and its effect on growth, photosynthesis and antioxidative responses were studied in Sesbania drummondii seedlings. Mercury concentration in shoots as well as in the roots increased with increasing Hg concentrations in the growth solution. The accumulation of Hg was more in roots than shoots. At 100 mg l-1 Hg concentration, shoots accumulated 998 mg Hg kg -1 dry weight (dw) while roots accumulated 41,403 mg Hg kg-1 dw. Seedlings growth was not significantly affected at lower concentrations of Hg. A concentration of 100 mg l-1 Hg inhibited growth by 36.8%, with respect to control. Photosynthetic activity was assessed by measuring chlorophyll a fluorescence by determination of Fv/Fm and Fv/Fo values. Photosynthetic integrity was not affected up to 50 mg l-1 Hg concentration, however, concentrations higher than 50 mg l-1 affected photosynthetic integrity. Sesbania responded to Hg induced oxidative stress by modulating non-enzymatic antioxidants [glutathione (GSH) and non-protein thiols (NPSH)] and enzymatic antioxidants: superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR). Glutathione content and GSH/GSSG ratio increased up to a concentration of 50 mg l-1 while slight down at 100 mg l-1 Hg. The content of NPSH significantly increased with increasing Hg concentrations in the growth medium. The activities of antioxidative enzymes, SOD, APX and GR followed the same trends as antioxidants first increased up to a concentration of 50 mg l-1 Hg and then slight decreased. The results of present study suggest that Sesbania plants were able to accumulate and tolerate Hg induced stress using an effective antioxidative defense mechanisms.     

Growth responses of birch and Sitka spruce exposed to acidified rain

Seedlings of birch and Sitka spruce were grown on a range of British soils for 2 years and exposed to simulated acid rainfall treatments of pHs 5.6, 4.5, 3.5 and 2.5. Both species developed visible leaf injury patterns when exposed to the pH 2.5 treatment. In Sitka spruce this leaf injury was followed by high needle loss during the first winter and greater mortality. Generally, height growth of Sitka spruce was unaffected by treatments, but acid rainfall at pH 2.5 increased the height of birch. Mean height of both species was strongly affected by soil type. Significant soils x treatment effects on the heights of both species indicated that on some soils plant growth responses to the treatments did not fit the general pattern. Hence, while the results indicate that generally ambient acidities of rainfall in the UK are unlikely to adversely affect the growth of birch or Sitka spruce, plants growing on some soils may be susceptible to injury.     

Comparison of photosynthetic responses to manganese toxicity of deciduous broad-leaved trees in northern Japan

The effects of manganese (Mn) toxicity on photosynthesis of four tree species in northern Japan representing different successional traits were examined. The four species are: Betula ermanii (Be) and Alnus hirsuta (Ah) representing two early successional species, Ulmus davidiana var. japonica (Ud) as the mid-successional species, and Acer mono (Am) as the late successional species. Seedlings were grown hydroponically in a solution containing nutrients and Mn of four concentrations (1, 10, 50, 100 mg litre(-1)) for 50 days. Gas exchange measurements indicate that in all species, Mn accumulation in leaves resulted in the decline of light-saturated net photosynthetic rate ai ambient CO(2) pressure (35 Pa, Pn(amb)) and at saturating (5%) CO(2) pressure (Pn(sat)), and of carboxylation efficiency but has little effect on the maximum efficiency of photochemistry. Sensitivity to elevated levels of Mn differed among species where the decline of Pn(amb) was much more modest in the two early successional species of Be and Ah than the mid- and late successional species of Ud and Am. The same trends were observed in both Pn(sat) and carboxylation efficiency. Based on these results, we suggest that early successional species (Betula ermanii and Alnus hirsuta) have greater tolerance for excess Mn in leaves than mid- and late successional species.     

Boron toxicity in the rare serpentine plant, Streptanthus morrisonii

The release of boron-laden mist from the cooling towers of some geothermal power stations in northern California potentially threatens nearby populations of the rare serpentine plant, Streptanthus morrisonii F. W. Hoffm. To assess the tolerance of S. morrisonii to high levels of boron, the effect of boron on leaf condition, life history, germination rate, growth rate, allocation and photosynthesis was measured on plants grown in a greenhouse. Relative to other species, S. morrisonii was tolerant of excess boron. On serpentine soil, mild to moderate toxicity symptoms (older leaves exhibiting chlorosis and necrosis, but few leaves killed) were apparent when the boron concentration in applied nutrient solutions was 240-650 microm. Severe toxicity symptoms (significant leaf loss, young leaves with toxicity symptoms) were apparent when the applied solution was over 1000 microm boron. Above 1000 microm boron, S. morrisonii appeared unable to complete its life cycle. On a tissue basis, boron toxicity was first observed when leaf boron content was 40-90 micromol g(-1) dry weight. In leaves with severe boron toxicity (> 35% injury), the boron content was generally above 130 micromol g(-1) dry weight. These levels were an order of magnitude above the tissue boron content of plants in the field. Prior to the onset of pronounced boron toxicity symptoms, growth rate, allocation patterns, and photosynthesis were unaffected by high boron. These results indicate that inhibition of growth and photosynthesis occurred because of a loss of viable tissue due to boron injury, rather than a progressive decline as leaf boron levels increased.     

Toxicity of uranium and copper individually, and in combination, to a tropical freshwater macrophyte (Lemna aequinoctialis)

Copper (Cu) and uranium (U) are of potential ecotoxicological concern to tropical freshwater biota in northern Australia, as a result of mining activities. Few data are available on the toxicity of U, and no data are available on the toxic interaction of Cu and U, to freshwater biota. This study determined the toxicity of Cu and U individually, and in combination, to a tropical freshwater macrophyte, Lemna aequinoctialis (duckweed), in a synthetic soft water (27 degrees C; pH, 6.5; hardness, 40 mg CaCO3 l-1, alkalinity, 16 mg CaCO3 l-1), typical of many fresh surface waters in coastal northern Australia. The growth rate of L. aequinoctialis decreased with increasing Cu or U concentrations, with the concentration of Cu inhibiting growth by 50% (EC50) being 16+/-1.0 microg l-1, with a minimum detectable effect concentration (MDEC) of 3.2 microg l-1. The concentration of U inhibiting growth by 50% (EC50) was 758+/-35 microg l-1 with a MDEC of 112 microg l-1. The EC50 value for the exposure of L. aequinoctialis to equitoxic mixtures of Cu and U was significantly (P0.05) higher than one toxic unit (1.35; 95% confidence interval, 1.18-1.52), indicating that the combined effects of Cu and U are less than additive (antagonistic). Therefore, inhibition of the growth rate of L. aequinoctialis was reduced when Cu and U were present in equitoxic mixtures, relative to individual metal exposures. Since non-additive (e.g. antagonistic) interactions of metal mixtures cannot be predicted using current mixture models, these results have important potential implications for the protection of freshwater ecosystems through the derivation of national water quality guidelines.     

Cadmium accumulation and its effect on the in vitro growth of woody fleabane and mycorrhized white birch

The effect of Cd on woody fleabane (Dittrichia viscosa (L.) Greuter) and white birch (Betula celtiberica Rothm. & Vasc.) was examined. Woody fleabane and white birch were grown in vitro in Murashige, T., Skoog, F., [1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473-479] (MS) plus Cd (10 mg Cd kg(-1)) and except for root length in white birch, plant development was inhibited when Cd was added. Cd accumulation in above-ground tissues showed differences among clones, reaching 1300 and 463 mg Cd kg(-1) dry wt. in selected clones of woody fleabane and white birch, respectively. Tolerance of Paxillus filamentosus (Scop) Fr. to Cd was also examined before mycorrhization. Plants of mycorrhized white birch grown in the presence of Cd had a better development and accumulated more Cd in their shoots than the non-mycorrhized ones. The use of selected clones of woody fleabane and the mycorrhization of white birch enhance extraction efficiency from contaminated soils in phytoremediation programs.     

The effects of simulated acid rain on the growth of three herbaceous species grown on a range of British soils

Seedlings of winter barley, perennial ryegrass and white clover were grown on a range of British soils for 21-24 weeks and exposed to simulated acid rainfall treatments of pHs 5.6, 4.5, 3.5 and 2.5. Whilst leaves of white clover developed leaf lesions after 18 weeks of exposure to the pH 2.5 treatments, there were no signs of visible injury to the other two species. At harvest, it was noted, for all species, that there was a large amount of variation in the sizes of individual plants and this made it difficult to detect differences between the treatments for plants on an individual soil. However, in combined analyses for all soils, it was found that the treatments had substantial effects on the yields of plants. The yields of both winter barley and clover were highly correlated with rainfall pH, showing substantial reductions in the more acid rains as compared to the pH 5.6 (control) treatment. In contrast, plants of perennial ryegrass produced higher yields of shoots at the most acid (pH 2.5) as compared to the other treatments. It was noted that the pH 2.5 treatment resulted in a generally lower soil pH at the termination of the experiment.     

Sources of errors in assessing ozone visible symptoms on native vegetation

This paper aims to identify the problems regarding the evaluation of ozone (and ozone-like) symptoms, by examining the results of the 4th UN/ECE ICP-Forests Intercalibration Course for the assessment of ozone-induced visible symptoms. Trees, shrubs and herbaceous species were evaluated in a tree nursery, at Lattecaldo (Switzerland) and under open field conditions at Moggio (Italy). The main findings were: (i) the most expert surveyors tended to be grouped in the same cluster and, during the field exercises, they tended to assess in a more conservative manner compared to the less trained participants; (ii) the agreement was greater in assessing the absence rather than the presence of symptoms; (iii) typical interveinal stippling on the upper leaf surface was more accurately evaluated than discoloration; (iv) uncertainties resulted mainly for species which showed greater variability in their symptom manifestation, and for certain herbaceous species.     

Visible and microscopic injury in leaves of five deciduous tree species related to current critical ozone levels

Because the current critical level of ozone (O(3)) for forest trees is based only on one species, the responses of five deciduous tree species were differentiated in a climate chamber experiment. The number of symptomatic leaves per tree was significantly increased, and stomatal conductance was decreased under 50% ambient+30 nl l(-1) O(3) as compared to 'normal' senescence at 50% ambient [O(3)]. Species with a high stomatal conductance did not show earlier or more leaf injury symptoms. The additional 30 nl l(-1) O(3) induced specific pectinaceous cell wall protrusions, phenolic cell wall incrustations, tonoplast vesicles, and inhomogeneous, condensed/precipitated phenolic material in the vacuoles. Due to added O(3), cell senescence was accelerated with increased electron-density of the cytoplasm, and initial chloroplast degeneration. The slow degeneration process started in mesophyll cells, and expanded into epidermal and finally guard cells. Because of the large variance in biomass between individuals and species, the current critical level is supported by the assessment of visible leaf symptoms rather than growth reduction.