Growth responses of three legume species exposed to simulated acid rain

Seedlings of Vicia faba L., Phaseolus multiflorus L. and Pisum sativum L. were raised during exposure to simulated acid rainfall treatments of pHs 5.6, 4.5, 3.5 and 2.5 at a rate of 30 mm per week. All three species were found to be adversely affected by the more acid pH 3.5 and pH 2.5 treatments after 7-8 weeks of exposure. There were total plant dry weight reductions of 40% for V. faba, 31% for P. sativum and 28% for P. multiflorus exposed to the pH 2.5 treatment, as compared to those grown in the control (pH 5.6 treatment). In addition, V. faba was found to be sensitive to the pH 4.5 treatment with an 18% reduction in total plant weights (compared to plants grown in the pH 5.6 treatment). In P. multiflorus, reduction in the dry weights of shoots in response to increasing acidity of rain was not accompanied by reduction in root weights, indicating an interference in the partitioning of assimilates. It is concluded that these three species, and V. faba in particular, may be growing below their potential in much of the UK.     

Stem growth reduction in mature Sitka spruce trees exposed to acid mist

An eighteen-year-old clone of Sitka spruce (Picea sitchensis (Bong.) Carr) growing in the field was used to evaluate the whole tree response of 'mature' Sitka spruce to acid mist treatment. The mist, an equimolar mixture of H(2)SO(4) and NH(4)NO(3) at pH 2.5 with or without particles (soda glass ballotini < 20 microm diameter), was applied twice weekly (equivalent to 4 mm precipitation week(-1)) throughout the growing season, May-November 1990-1992. The annual dose of S, N, H applied as mist (at 51, 48 and 3.3 kg ha(-1), respectively) was 2.5 times that measured in the Scottish uplands. Throughout the experiment there was no evidence of visible injury symptoms, yet there was a highly significant reduction (p < 0.02) in the stem-area increment relative to the stem area at the start, measured using vernier dendrometer bands. There was no significant difference between the (acid mist + particle) and the acid mist only treatments. The mean relative stem-area increment over two complete growing seasons (1991-1992) was 65% for control trees, but only 53% for acid-misted trees.     

Antioxidant responses to simulated acid rain and heavy metal deposition in birch seedlings

This study measured the responses of different anti-oxidants in 2-year-old birch (Betula pendula Roth) seedlings subjected to simulated acid rain (pH 4.0) and heavy metals (Cu/Ni), applied alone or in combination for 2 months. The applied concentrations of pollutants did not significantly affect seedling biomass or total glutathione levels. Acid rain alone increased superoxide dismutase (SOD) activity both in leaves and roots, while heavy metals alone inhibited SOD activity in roots. Both acid rain and heavy metals applied singly increased ascorbate peroxidase (APX) and guaiacol peroxidase (GPX) activities in leaves but decreased activities in roots. In contrast, acid rain and heavy metal treatments increased glutathione reductase (GR) activity in roots but not in leaves. Spraying birch seedlings with a mixture of acid rain and heavy metals increased SOD, APX and GPX activities in leaves and GR activity in roots. However, the effects of mixed pollutants on enzyme activities usually were less than the summed effects of individual pollutants. Enzyme responses also depended on where pollutants were applied: spraying pollutants onto the shoots initiated higher responses in SOD, APX and GPX than did application to the soil surface, while the opposite was true for GR.     

Effects of nitrogen with and without acidified sulphur on an ectomycorrhizal community in a Sitka spruce (Picea sitchensis Bong. Carr) forest

This preliminary study investigated the effects of enhanced nitrogen (NH4NO3 at 48 kg ha(-1) y(-1)), sulphur (Na2SO4 at 50 kg ha(-1) y(-1)), acidified nitrogen and sulphur (H2SO4 + NH4NO3) at pre-stated doses (pH 2.5), and acidified nitrogen and sulphur deposition at double these doses on the ectomycorrhizal community associated with a 13-year-old Sitka spruce (Picea sitchensis) forest. Sulphur deposition had little impact on below ground ectomycorrhizal diversity, but stimulated sporocarp production. Nitrogen inputs increased below ground colonisation compared to acidified nitrogen and sulphur, largely due to an increase in Tylospora fibrillosa colonisation. Sporocarp production and ectomycorrhizal root colonisation by Lactarius rufus were reduced in the nitrogen treated plots. These observations suggest that nitrogen deposition to a young plantation may suppress ectomycorrhizal fungi producing large sporocarps. It is proposed that enhanced nitrogen deposition increases ectomycorrhizal nitrogen assimilation, consuming more carbon and leaving less for extrametrical mycelium and sporocarp development.     

Interactions between Sitka spruce, the green spruce aphid, sulphur dioxide pollution and drought

Sitka spruce trees, with and without the aphid Elatobium abietinum and/or drought treatment, were subjected to 25 nl litre(-1) of sulphur dioxide over a 2-month period. Aphids became three times as abundant on the fumigated trees if they were well watered and twice as abundant on trees from which water was withheld, compared with unfumigated controls. Growth parameters of the trees were little affected by pollution alone, but were substantially reduced by either aphids or drought. There was a significant interaction between SO(2) and aphids in a further reduction of both leader extension and root weight. Root weight was also reduced by 24% more than expected from the additive effects of the combined SO(2) and drought treatment.     

Mycorrhizal populations and fine root development on Norway spruce exposed to controlled doses of gaseous pollutants and simulated acidic rain treatments

Effects after long-term fumigation and simulated acidic rain (pH 4.0) treatment in open-top chambers (OTC) during 5 vegetation periods on ectomycorrhizal (EM) frequency, fine root structure and distribution of short roots were recorded for Norway spruce. Simultaneous application of ozone and SO(2) resulted in a significant decrease (P< or =0.01) in the proportion of EM root tips and an increase in the numbers of necrotic short roots. The observed differences were partly due to more than additive effects of ozone and SO(2). On the basis of EM and non-mycorrhizal (NM) short roots in the irrigated controls (OTC-V, pH 5.6, Ah-horizon: EM + NM/cm=0.05) the acid treatments resulted in drastic effects (P< or =0.05) on the branching density and the proportion of EM root tips and NM short roots. Relative numbers of short roots infected by indigenous EM fungi with trees in the fumigation treatments at pH 4.0 were similar: 1.5-1.9 EM tips/cm and differed noticeably from the control: 5.2 EM tips/cm. Again, increasing numbers of necrotic and NM short roots were noted. Parallel measurements by the ingrowth-core technique confirm the argument, that EM formation seems to be a reliable indicator of pollution-induced stress.     

Open-top chamber and field exposure of Sitka spruce to simulated acid mist: a comparison of results

Sitka spruce seedlings grown in pots of compost in open-top chambers (OTCs) or outside, in an agricultural loam soil, in different years, were treated with pH 2.5 acid mist comprising an equimolar solution, of H₂SO₄ and NH₄NO₃ at 1.6 mol m⁻³. The effects of rain, and frequency of acid mist applications on visible injury, nutrition and frost hardiness were evaluated in the OTC experiment. Similar evaluations were undertaken in the field where application frequency was dependent on windspeed, rain and direct sunlight. Rain washing significantly ameliorated the effect of acid mist on frost hardiness in OTCs. In the field experiment there was no effect of acid mist treatment. Increasing the frequency of misting in OTCs for the same exposure exacerbated the detrimental effects of acid mist, causing both visible damage and further increasing the sensitivity of the acid treated trees to freezing temperatures. The results show that the treatment environment can both ameliorate and exacerbate treatment effects.     

Anatomical and physiological responses of Colorado blue spruce to vehicle exhausts

In order to examine whether the leaves of the Colorado blue spruce (Picea pungens) are damaged or not by traffic pollution, the traits of the anatomy and physiology of its leaves are investigated by exposure to vehicle exhausts in a laboratory experiment lasting 30 days. The results show that both the anatomical structures and physiological traits of the leaves are significantly affected by vehicle exhausts. The anatomical structures, including epidermis, cuticle, palisade, and spongy parenchyma are modified when exposed to the high concentrations (≥0.4 mg/m³) of vehicle exhausts. However, physiological traits such as total chlorophyll content are not changed when exposed to different concentrations of vehicle exhaust. Unlike the total chlorophyll content, the electrical conductivities increased, whereas the POD activities decreased when presented in vehicle exhausts. The present study indicates that the Colorado blue spruce changes its anatomical structures and physiological traits to avoid possible damage by vehicle exhausts.     

Indication of airborne pollution by birch and spruce in the vicinity of copper smelter

The aim of the study was determination of air pollution impact of the copper smelter in Bor and its surroundings (Serbia) by assessing the suitability of birch (Betula pendula Roth.) and spruce (Picea abies L.) for the purposes of biomonitoring and comparing it with previously published data from the same study area. The concentrations of Cu, Zn, Pb and Mn in leaves/needles, branches, roots and soil were determined. Sampling was performed during 2009 in two zones with high load of air pollution due to copper mining and smelting activities, and one background zone. Metal accumulation and translocation was evaluated in terms of biological factors. In addition, plant enrichment factor was calculated. According to the results, plant foliage was not enriched through soil, which indicates absorption from the air, with both species acting as excluders of Cu, Pb, Zn and Mn. Leaves were more enriched with all the metals than needles, indicating a better response of birch to airborne pollution than spruce. Cluster analysis showed different level of pollution at the sites, while correlations between Cu and Pb obtained by Principal Component Analysis indicated their anthropogenic origin. Regarding previously published results, beside birch leaves, pine needles (which showed higher level of response to pollution compared to linden leaves) could be applied in air biomonitoring surveys near copper smelters.     

Biological responses to the chemical recovery of acidified fresh waters in the UK

We report biological changes at several UK Acid Waters Monitoring Network lakes and streams that are spatially consistent with the recovery of water chemistry induced by reductions in acid deposition. These include trends toward more acid-sensitive epilithic diatom and macroinvertebrate assemblages, an increasing proportional abundance of macroinvertebrate predators, an increasing occurrence of acid-sensitive aquatic macrophyte species, and the recent appearance of juvenile (<1 year old) brown trout in some of the more acidic flowing waters. Changes are often shown to be directly linked to annual variations in acidity. Although indicative of biological improvement in response to improving water chemistry, "recovery" in most cases is modest and very gradual. While specific ecological recovery endpoints are uncertain, it is likely that physical and biotic interactions are influencing the rate of recovery of certain groups of organisms at particular sites.     

Effects of ozone and simulated acid rain on birch seedling growth and formation of ectomycorrhizae

Four-week-old paper birch (Betula papyrifera Marsh.) seedlings, inoculated or non-inoculated with the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker & Couch and grown in steamed or non-steamed soil, were exposed to ozone (O(3)) and/or simulated acid rain (SAR). Plants were exposed to O(3) for 7 h per day on 5 days per week for 12 weeks. O(3) concentrations were maintained between 0.06 and 0.08 ppm. SAR was applied 10 min per day on 2 days per week. O(3), SAR, soil regime and mycorrhizal treatment did not significantly affect any of the measured variables. Interactions between O(3) and SAR, SAR and mycorrhizal treatment, soil regime and mycorrhizal treatment and ozone and soil regime had significant effects. Treatment of seedlings with pH 3.5 SAR caused increases in growth which were more apparent in birch exposed to O(3). Mucorrhizal treatment caused increased growth in non-steamed soil, while growth appeared to decrease in steamed soil. Birch seedlings grew much better in steamed soil. The implications of increased growth in steamed soil may demonstrate the importance of looking at the secondary effects of pollutants on soil-borne organisms.     

Trichloroacetic acid cycling in Sitka spruce saplings and effects on sapling health following long term exposure

Trichloroacetic acid (TCA, CCl(3)COOH) has been associated with forest damage but the source of TCA to trees is poorly characterised. To investigate the routes and effects of TCA uptake in conifers, 120 Sitka spruce (Picea sitchensis (Bong.) Carr) saplings were exposed to control, 10 or 100 microg l(-1) solutions of TCA applied twice weekly to foliage only or soil only over two consecutive 5-month growing seasons. At the end of each growing season similar elevated TCA concentrations (approximate range 200-300 ng g(-1) dwt) were detected in both foliage and soil-dosed saplings exposed to 100 microg l(-1) TCA solutions showing that TCA uptake can occur from both exposure routes. Higher TCA concentrations in branchwood of foliage-dosed saplings suggest that atmospheric TCA in solution is taken up indirectly into conifer needles via branch and stemwood. TCA concentrations in needles declined slowly by only 25-30% over 6 months of winter without dosing. No effect of TCA exposure on sapling growth was measured during the experiment. However at the end of the first growing season needles of saplings exposed to 10 or 100 microg l(-1) foliage-applied TCA showed significantly more visible damage, higher activities of some detoxifying enzymes, lower protein contents and poorer water control than needles of saplings dosed with the same TCA concentrations to the soil. At the end of each growing season the combined TCA storage in needles, stemwood, branchwood and soil of each sapling was <6% of TCA applied. Even with an estimated half-life of tens of days for within-sapling elimination of TCA during the growing season, this indicates that TCA is eliminated rapidly before uptake or accumulates in another compartment. Although TCA stored in sapling needles accounted for only a small proportion of TCA stored in the sapling/soil system it appears to significantly affect some measures of sapling health.     

Foliage responses of spruce trees to long-term low-grade sulfur dioxide deposition

Foliage on spruce trees (Picea rubens Sarg.) growing on dry SO(2) deposition zones (dry SO(2) deposition ranging from 0.5 and 8.5 S kg ha(-1) year(-1)) downwind from a SO(2) emission source was analyzed to assess chronic effects of long-term low-grade SO(2) deposition on net photosynthesis, stomatal conductance, dark respiration, stomatal antechamber wax structures, elemental concentrations in and on foliage (bulk and surficial concentrations), and types of epiphytic fungi that reside in the phylloplane. Elemental distributions on stomatal antechambers, on fungal colonies, and on smooth surfaces between stomates and fungus colonies were determined with a scanning electronic microscope (SEM) by way of X-ray scanning. It was found that net photosynthesis of newly developed spruce foliage (current-year, and 1-year-old) was not significantly affected by the local SO(2) deposition rates. Sulfur dioxide deposition, however, may have contributed to the gradual decrease in net photosynthesis with increasing needle age. Dark respiration rates were significantly higher on foliage taken from high SO(2) deposition zones. Stomatal rod-web structures deteriorated to flakes with increasing needle age and increasing SO(2) deposition. Further inspection of the needle surfaces revealed an increasing abundance of fungal colonies with increasing needle age. Many fungal taxa were isolated and identified. It was found that black yeasts responded positively, and Xylohypha pinicola responded negatively to high rates of SO(2) deposition. Surficial concentrations of elements such as P, S, K, Cl, Ca were about 10 times higher on fungal colonies than on smooth needle surfaces. Surficial Ca contents on 4 or 5-year-old needles decreased with increasing SO(2) deposition, but surficial S concentrations remained the same. In contrast, bulk foliar Ca and S concentrations increased with increasing SO(2) deposition.     

Anchoring novel molecular biomarker responses to traditional responses in fish exposed to environmental contamination

The responses of Dicentrarchus labrax and Liza aurata to aquatic pollution were assessed in a contaminated coastal lagoon, using both traditional and novel biomarkers combined. DNA damage, assessed by comet assay, was higher in both fish species from the contaminated sites, whereas levels of cytochrome P450 1A1 gene expression were not significantly altered. The liver histopathological analysis also revealed significant lesions in fish from contaminated sites. Alterations in ras and xpf genes were analysed and additional pollutant-responsive genes were identified. While no alterations were found in ras gene, a downregulation of xpf gene was observed in D. labrax from a contaminated site. Suppression subtractive hybridization applied to D. labrax collected at a contaminated site, revealed altered expression in genes involved in energy metabolism, immune system activity and antioxidant response. The approach and results reported herein demonstrate the utility of anchoring traditional biomarker responses alongside novel biomarker responses.     

Throughfall chemistry and canopy interactions in a Sitka spruce plantation sprayed with six different simulated polluted mist treatments

Throughfall chemistry was studied in a mature Sitka spruce plantation in order to investigate canopy interactions, such as nitrogen absorption, cation leaching, and neutralization of rainfall passing through the canopy. The plantation had been exposed to six different simulated mist treatments including N (NH(4)NO(3)) and S (H(2)SO(4) at pH 2.5) in four replicated blocks since 1996. Throughfall and rainfall were collected from May to September 2000. The results showed that 30-35% of the applied N was retained by the canopy. There were linear relationships between the loss of H(+) and increased K(+), Mg(2+) and Ca(2+) deposition through the canopy. However these increases in K(+), Mg(2+) and Ca(2+) deposition accounted for only about 50% of total neutralization of the acidity. The relationship between the anion deficits in throughfall and the loss of H(+) implied that weak organic acid anions were involved in the neutralization of the acidity in throughfall.     

Effects of acid mist on mature grafts of Sitka spruce. Part I. Frost hardiness and foliar nutrient concentrations

Mature grafts of five clones of Sitka spruce (Picea sitchensis Bong. Sarg.) were exposed to simulated acid mist composed of an equimolar mixture of sulphuric acid and ammonium nitrate at pH 2.5 and pH 5.0 in open-top chambers from May to November 1991. Treatments were applied on consecutive days, four times a week. The pH 2.5 treatment provided an overall dose three times higher than that received by forests in upland areas of Britain. Frost hardiness was assessed in November by freezing detached current year shoots at a range of temperatures and assessing the rate of electrolyte leakage Foliar nutrient concentrations were determined on the same shoots. Acid mist at pH 2.5 significantly reduced frost hardiness in four of the five clones; the temperature causing 50% shoot death (LT50) was increased by 0 to 7 degrees C. The clones varied in their level of hardiness, one clone being exceptionally frost sensitive. The frost hardiness of the frost sensitive clone was found to be less perturbed by acid mist than the hardiness of the more frost resistant clones. Mature grafts showed a smaller reduction in hardiness at an equivalent dose than that found previously with Sitka spruce seedlings. Compared with seedlings, grafts had lower absolute concentrations of foliar sulphur. Exposure to acid mist at pH 2.5 increased %S in current year foliage by <0.05% compared with absolute increases of more than 0.10% in current year foliage of seedlings. We conclude that the effect of acid mist on frost hardiness is likely to be less on mature trees than on seedlings and that the increased frost risk to mature trees of Sitka spruce from occult deposition alone is small.     

Relationship between foliar injury and changes in antioxidant levels in red and Norway spruce exposed to acidic mists

Elevated levels of total glutathione and enhanced activities of glutathione reductase and ascorbate peroxidase were found in needles of red spruce which had been exposed to acidic mists. Reduced levels of ascorbate were also detected. Such observations suggest that oxidative stress is involved in processes which resist foliar injury caused by acidic misting. Different ionic compositions of the acidic mists applied had pronounced effects on the levels of these antioxidants and the activities of related enzymes. Sulphate was found to be most effective in causing increases in glutathione, while additions of ammonium and/or nitrate mitigated the effects of sulphuric acid. Moreover, it is the composition of ions in the applied mists, rather than the levels of acidity, that determines the extent of the overall response of red spruce. By contrast, although acidic mists caused similar increases in glutathione content of Norway spruce needles, no statistically significant changes of ascorbate or related enzymic activities were found.     

Effects of simulated rain acidity on ectomycorrhizae of red spruce seedlings potted in natural soil

Formation of ectomycorrhizae of red spruce (Picea rubens) grown in natural soil was measured after seedlings were exposed to 25 or 50 applications of simulated rain of pH 5.5, 3.5 or 2.5. Ectomycorrhizae were quantified as the total number of ectomycorrhizal tips per centimeter, and as the number of ectomycorrhizal tips for each morphotype and for Cenococcum geophilum. Rain solutions were applied to the soil alone, to foliage and stem alone, or to entire potted seedlings. Final soil pH was linearly related to rain solution acidity. Lower base saturation, calcium and zinc content, and higher exchangeable acidity were observed after pH 2.5 treatments if the soil was exposed. Rain solutions and the subsequent changes in soil characteristics did not affect the total numbers of ectomycorrhizal tips. Four morphotypes of ectomycorrhizae observed for these seedlings were unaffected by simulated rain. However, the numbers of ectomycorrhizal tips formed by C. geophilum tended to increase with rain solution acidity after 50 applications. Method of rain deposition did not affect ectomycorrhizae, suggesting both plant and soil mediated responses may favor certain mycobionts. The results of this study indicate that short-term acidic deposition does not induce significant changes in the frequency of ectomycorrhizae, but higher numbers of C. geophilum tips suggest there may be changes in the relative occurrence of specific morphotypes of fungus species.     

Growth and photosynthetic responses to copper in wild grapevine

The present study evaluates the tolerance and accumulation potential of Vitis vinifera ssp. sylvestris under moderate and high external Cu levels. A greenhouse experiment was conducted in order to investigate the effects of a range of external Cu concentrations (0–23 mmol L⁻¹) on growth and photosynthetic performance by measuring gas exchange, chlorophyll fluorescence parameters and photosynthetic pigments. We also measured the total copper, nitrogen, phosphorus, sulphur, calcium, magnesium, iron, potassium and sodium concentrations in the plant tissues. All the experimental plants survived even with external Cu concentrations as high as 23 mmol L⁻¹ (1500 mg Cu L⁻¹), although the excess of metal resulted in a biomass reduction of 35%. The effects of Cu on growth were linked to a reduction in net photosynthesis, which may be related to the effect of the high concentration of the metal on photosynthetic electron transport. V. vinifera ssp. sylvestris survived with leaf Cu concentrations as high as 80 mg kg⁻¹ DW and growth parameters were unaffected by leaf tissue concentrations of 35 mg Cu kg⁻¹ DW. The results of our study indicate that plants of V. vinifera ssp. sylvestris from the studied population are more tolerant to Cu than the commercial varieties of grapevine that have been studied in the literature, and could constitute a basis for the genetic improvement of Cu tolerance in grapevine.