Physiological effects of long term exposure to low concentrations of SO2 and NH3 on poplar leaves

Shoots of poplar (Populus euramericana L. cv. Flevo) were exposed to filtered air, SO₂, NH₃ or a mixture of SO₂ and NH₃ for 7 weeks in fumigation chambers. After this exposure gas exchange measurements were carried out using a leaf chamber. As compared to leaves exposed to filtered air, leaves pretreated with 112 μg m^?3 SO₂ showed a small reduction in maximum CO₂ assimilation rate (P_max) and stomatal conductance (g_s). They also showed a slightly higher quantum yield and dark respiration. In addition, the fluorescence measurements indicated that the Calvin cycle of the leaves pretreated with 112 μg m^?3 SO₂ was more rapidly activated after transition from dark to light. An exposure to 64 μg m^?3 NH₃ had a positive effect on P_max, stomatal conductance and NH₃ uptake of the leaves. This positive effect was counteracted by an SO₂ concentration of 45 μg m^?3. The exposure treatments appeared to have no effect on the relationship between net CO₂-assimilation and g_s. Also, no injury of the leaf cuticle or of epidermal cells was observed. Resistance analysis showed that NH₃ transfer into the leaf can be estimated from data on the boundary layer and stomatal resistance for H₂O transfer and NH₃ concentration at the leaf surface, irrespective of whether the leaves are exposed for a short or long time to NH₃ or to a mixture of NH₃ and SO₂. In contrast SO₂ uptake into the leaves was only partly correlated to the stomatal resistance. The results suggest a large additional uptake of this gas by the leaves. The possibility of a difference in path length between SO₂ and H₂O molecules is proposed.     

Natural CO2 springs in Italy: a resource for examining long-term response of vegetation to rising atmospheric CO2 concentrations

It is estimated that more than 100 geothermal CO₂ springs exist in central-western Italy. Eight springs were selected in which the atmospheric CO₂ concentrations were consistently observed to be above the current atmospheric average of 354μmol mol^-1. CO₂ concentration measurements at some of the springs are reported. The springs are described, and their major topographic and vegetational features are reported. Preliminary observations made on natural vegetation growing around the gas vents are then illustrated. An azonal pattern of vegetation distribution occurs around every CO₂ spring regardless of soil type and phytoclimatic areas. This is composed of pioneer populations of a Northern Eurasiatic species (Agrostis canina L.) which is often associated with Scirpus lacustris L. The potential of these sites for studying the long-term response of vegetation to rising atmospheric CO₂ concentrations is discussed.     

The impact of elevated CO2 on growth and photosynthesis in Agrostis canina L. ssp. monteluccii adapted to contrasting atmospheric CO2 concentrations

 The aim of this study was to characterise growth and photosynthetic capacity in plants adapted to long-term contrasting atmospheric CO₂ concentrations (C _a). Seeds of Agrostis canina L. ssp. monteluccii were collected from a natural CO₂ transect in central-western Italy and plants grown in controlled environment chambers at both ambient and elevated CO₂ (350 and 700 μmol mol⁻¹) in nutrient-rich soil. Seasonal mean C _a at the source of the plant material ranged from 610 to 451 μmol CO₂ mol⁻¹, derived from C₄ leaf stable carbon isotope discrimination (δ¹³C). Under chamber conditions, CO₂ enrichment stimulated the growth of all populations. However, plants originating from elevated C _a exhibited higher initial relative growth rates (RGRs) irrespective of chamber CO₂ concentrations and a positive relationship was found between RGR and C _a at the seed source. Seed weight was positively correlated with C _a, but differences in seed weight were found to explain no more than 34% of the variation in RGRs at elevated CO₂. Longer-term experiments (over 98 days) on two populations originating from the extremes of the transect (451 and 610 μmol CO₂ mol⁻¹) indicated that differences in growth between populations were maintained when plants were grown at both 350 and 700 μmol CO₂ mol⁻¹. Analysis of leaf material revealed an increase in the cell wall fraction (CWF) in plants grown at elevated CO₂, with plants originating from high C _a exhibiting constitutively lower levels but a variable response in terms of the degree of lignification. In vivo gas exchange measurements revealed no significant differences in light and CO₂ saturated rates of photosynthesis and carboxylation efficiency between populations or with CO₂ treatment. Moreover, SDS-PAGE/ LISA quantification of leaf ribulose bisphosphate carboxylase/oxygenase (Rubisco) showed no difference in Rubisco content between populations or CO₂ treatments. These findings suggest that long-term adaptation to growth at elevated CO₂ may be associated with a potential for increased growth, but this does not appear to be linked with differences in the intrinsic capacity for photosynthesis. Received: 16 August 1996 / Accepted: 19 October 1996     

Growth of coniferous trees exposed to SO2 and O3 using an open-air fumigation system

Scots pine (Pinus sylvestris L.), Norway spruce (Picea abies (L.) Karst.) and Sitka spruce (Picea sitchensis Bong. Carr.) were planted as 2-year-old seedlings in an open-air fumigation facility at Liphook in southern England in March 1985. The soil was a humoferric podzol of pH 4. SO₂ fumigation began in May 1987 and continued until December 1990. Long-term mean SO₂ concentrations were 4,13 and 22 nmol mo^?1. Three plots, one at each SO₂ level, were also exposed to O₃ at an average of 1–3.times the ambient level. O₃ fumigation ran from March to December 1988, May to December 1989 and February to December 1990. Each species reacted differently to treatment. Scots pine showed no growth response to either pollutant, although other work on the site demonstrated a number of deleterious effects of SO₂ on this species, including increased leaf loss and foliar injury. Stem basal diameter growth of Norway spruce was depressed in SO₂-treated plots. In contrast, extension growth of shoots of Sitka spruce increased in SO₂-treated plots, in apparent response to codeposition of NH₃-N. However, diameter growth of Sitka spruce main stems did not increase. No effects of O₃ on growth were recorded for any species.     

Effects of SO2 on photosynthesis and ribulose bisphosphate carboxylase in pine tree seedlings

Fumigations of 4–7 week old seelings of Pinus silvestris , with ca 400 μg SO₂ m^-3 for 4–5 days resulted in a decrease both in net photosynthetic rate and in the levels of ribulose bisphosphate carboxylase activity of the primary needles. The effect on net photosynthesis was reversible. No visible damages were observed on the needles. Lower concentrations, ca 200 μg SO₂ m^-3, for ca 6 days inhibited net photosynthesis reversibly. The effect on net photosynthesis was not the result of stomata closure or of increased respiration. A dose-response relation for the inhibition of net photosynthesis and ribulose bisphosphate carboxylase is suggested. A similar relation was observed for the chlorophyll content of the needles. The reversible inhibition of photosynthesis was evident only at high quantum flux density, and the quantum yield of photosynthesis at rate limiting quantum flux was not affected. This was also suggested from an experiment with Scots pine in the field.     

The effect of open-air fumigation with SO2 and O3 on carbohydrate metabolism in Scots pine (Pinus sylvestris) and Norway spruce (Picea abies)

The carbohydrate metabolism of the needles of Scots pine (Pinus sylvestris) and Norway spruce (Picea abies) has been examined in trees that were exposed to SO₂, and O₃, in an open-air fumigation experiment located in the Liphook forest in southern England. Two-year-old seedlings were planted in 1985 in seven experimental plots. Five plots received fumigation treatments of SO₂, O₃ or a combination of these gases to give a 2 × 3 factorial design with one additional ambient plot Fumigation with SO₂, occurred from May 1987 to December 1990 and O₃, fumigation occurred from March to December 1988, May to December 1989 and February to December 1990. Five samples of needles for investigation of carbohydrate metabolism were taken between February and July 1989. The concentrations of soluble carbohydrates (including sucrose and hexoses) were greatly reduced in the needles taken from Scots pine growing in the treated plots, and were also reduced, but to a lesser extent, in the needles taken from Norway spruce. Little variation in the concentration of starch in the needles of either species was detected. The activities of the two final enzymes of sucrose synthesis, sucrose phosphate synthase and sucrose 6-phos-phate phosphatase, were greatly reduced in the needles of Scots pine and were also reduced, but to a lesser extent, in the needles of Norway spruce in the fumigated plots. These reductions could be correlated with decreases in rates of photosynthetic CO₂ assimilation determined by independent groups of researchers working on the Liphook site.     

An in vivo analysis of the effect of SO2 fumigation on photosynthesis in Zea mays.

Fumigation of leaves with SO₂ can reduce the capacity for photosynthetic CO₂ uptake even in the absence of visible symptoms of damage. In vitro studies suggest that this invisible injury to intact leaves could be affected by damage to each of the main stages in the photosynthetic process. Reduced stomatal apertures may also reduce photosynthesis following SO₂ fumigation. The responses of CO₂ uptake by leaves to intercellular CO₂ concentration and to absorbed light provide information for quantitative separation of the in vivo contribution of the different stages of photosynthesis to reduction in overall rate. This study uses these techniques to examine the basis of reduction in CO₂ uptake in Zea mays cv. LG11 leaves following short-term fumigation with SO₂. Fumigation with 33 μmol m^–3 SO₂ for 30 min reduced light saturated CO₂ uptake by about one-third. An even greater reduction in light limited CO₂ uptake was observed and with no significant change in light absorptance this was attributed to a reduced quantum yield of photosynthesis. The light saturated CO₂ uptake rate and the stomatal conductance decreased in parallel. However, the relationship of CO₂ uptake to the intercellular CO₂ concentration suggested that the reduced stomatal conductance did not account for the reduced rate of CO₂ uptake following fumigation. Both the initial slope and plateau of this relationship were significantly reduced, suggesting that both carboxylation efficiency and capacity for regeneration of CO₂ acceptor were diminished by SO₂ fumigation. The operating intercellular CO₂ concentration indicated that both processes were co-limiting, before and after fumigation. The time required for induction of photosynthetic CO₂ uptake on illumination was approximately doubled following SO₂ fumigation, showing that fumigation impairs the ability of the photosynthetic apparatus to adapt to fluctuations in light level.     

Photosynthetic performance, chloroplast pigments and mineral content of Norway spruce (Picea abies (L.) Karst.) exposed to SO2 and O3 in an open-air fumigation experiment

Photosynthetic performance, mineral content and chloroplast pigments were investigated in August-September 1988 and 1989 in Norway spruce trees (Picea abies (L.) Karst.) exposed to SO₂, and O₃ in an open-air fumigation facility at Liphook, England. The data do not suggest a treatment effect on the mineral content of the needles in terms of nutrient leaching from the foliage. In addition, there were no direct SO₂ and/or O₃ effects on the content and/or composition of the chloroplast pigments. However, the long-term application of SO₂ resulted in a depression of net photosynthesis under light saturation and ambient CO₂ (A ₃₄₀) which was probably caused by a treatment-related depression of the carboxylation efficiency (CE). In 1989, the supposed treatment effects were apparently masked by an insufficient N-supply and probably also by low water availability during summer. However, fumigation appeared to accelerate an N-deficiency-related decrease of CE, stomatal closure and the age-dependent development of the chlorophyll content of the needles. In 1989, an observed depression of the photosynthetic capacity (A₂₅₀₀) was in part accompanied by a decrease in light use efficiency (α), suggesting an enhanced photosensitivity resulting from the impact of several possible interacting stresses (drought, N deficiency and fumigation). The results support the general conclusion that long-term low-level SO₂ dosage adversely affects the photosynthetic performance of the needle, whether directly or indirectly, and may also interact with other environmental stresses. The findings of our investigations are discussed with regard to the hypothesis of forest decline in the mountain regions of the Fichtelgebirge (north-eastern Bavaria, Germany).     

Antioxidant systems in the leaf apoplast compartment of Pinus pinaster Ait. and Pinus radiata D. Don. Plants exposed to SO2

The activities of guaiacol peroxidase (GuPOD), ascorbate peroxidase (ASAp), superoxide dismutase (SOD) and ascorbate/glutathione cycle (AGC) enzymes, together with ascorbate (ASC) and glutathione contents, were determined in apoplastic-fluid and cell-wall fractions of needles of Pinus pinaster Ait. and Pinus radiata D. Don. exposed for up to 6 months to SO₂ (0.01 ppm or 0.30 ppm) in fumigation chambers. AGC enzyme activities (monodehydroascorbate reductase, dehydroascorbate reductase and glutathione reductase) were in all cases undetectable, as was glutathione content. In needles of P. pinaster plants exposed to SO₂, ascorbate content and all enzyme activities considered (except AGC enzymes) increased. The increases were most marked in response to the higher SO₂ concentration. In needles of P. radiata, similar but less marked responses were observed. These findings suggest a) that enzyme activities and ascorbate contents increase in order to deal with the reactive oxygen intermediates produced during long-term contamination with SO₂, and b) that P. pinaster has more effective defences against contamination of this type than P. radiata.     

SO2-dependent cation competition and compartmentalization in Norway spruce needles

Ion contents in needles from Norway spruce trees [Picea abies (L.) Karst.] growing in Würzburg and in the SO₂-polluted Erzgebirge mountains were analysed to quantify cations which accumulate together with sulphate. In Würzburg there was a positive correlation of potassium (0.680 ± 0.300 Eq Eq^?1 SO₄^?2), magnesium (0.415 ± 0.111 Eq Eq^?1 SO₄^?2) and zinc (0.059 ± 0.006 Eq Eq^?1 SO₄^2?). In the Erzgebirge, potassium was also the stoichiometrically most important cation (0–887 ± 0–180 Eq K⁺ Eq^?1 SO₄^2?). All other correlations examined were weak or statistically non-significant. At both sites the calcium content of spruce needles did not depend on the sulphate content. The lack of a role for Ca²⁺ in neutralizing sulphate is a consequence of the presence of free oxalic acid in needles. Soluble oxalic acid precipitates Ca²⁺, which thereby becomes unavailable as a counterion for SO₄^2?. The activity coefficients of Ca²⁺ and oxalate^2?, and the solubility product of Ca-oxalate, were determined from in vivo data. It is concluded that the chronic accumulation of atmospheric sulphate in spruce needle vacuoles depletes available potassium and thereby strongly interferes with spruce growth and canopy turnover. This leads to impaired spruce vitality, even at sites where acute SO₂ disease symptoms are absent.