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.     

Persistent effects of low concentrations of SO2 and NO2 on photosynthesis in Scots pine (Pinus sylvestris) needles

In an open-field experiment, 50-year-old trees of Scots pine (Pinus sylvestris L.) were fumigated with low concentrations of SO₂ and NO₂ (10–15 nl I^?1) during the growing season in four consecutive years (1988 to 1991). Results from the autumn and early winter of 1991 and 1992 are presented. The maximum photochemical efficiency of photosystem II (PSII), as indicated by the ratio of variable to maximum fluorescence (F_v/F_M) was assessed in current and one-year-old needles from the top and the bottom of the canopy. Furthermore, simultaneous measurements of photosynthetic O₂ evolution and chlorophyll fluorescence were made in current-year needles at 20°C. In general, the F_v/F_M ratio as well as the gross rate of O₂ evolution in needles of fumigated trees was not significantly different from that in needles of control trees during the fumigation period. However, both current and one-year-old needles sampled in November and December 1991 from the top of the canopy of fumigated trees had significantly lower F_v/F_M values than corresponding needles of control trees. Similar differences in F_v/F_M correlated with the treatments were observed in needles from the bottom of the canopy, indicating that the depression of F_v/F_M in needles of fumigated trees was not due to an increased susceptibility to photoinhibition. In 1992, when no fumigation occurred, differences in F_v/F_M between the treatments were not significant during autumn and early winter. The gross rate of O₂ evolution at high irradiances was significantly lower in current-year needles of fumigated trees sampled in November and December 1991 than in those of control trees. Furthermore, a nearly identical linear relationship between the quantum yield of PSII electron transport determined from chlorophyll fluorescence and the quantum yield of O₂ evolution (gross rate of O₂ evolution/PPFD) was found during autumn and early winter. This appeared to be largely a result of changes in the thermal energy dissipation within PSII. The observed differences in photosynthetic characteristics correlated with the different treatments after the fumigation period is suggested to be mainly caused by increased sensitivity of the needles of fumigated trees to low and subfreezing temperatures. However, current-year needles of fumigated trees tended to have a lower N content than those of control trees, which may partly explain the differences in gross photosynthesis between fumigated and control trees.     

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.     

The effects of SO2 and O3 on the foliar nutrition of Scots pine, Norway spruce and Sitka spruce in the Liphook open-air fumigation experiment

Foliar elements were analysed in Scots pine, Sitka spruce and Norway spruce over a 6 year period before and during continuous exposure to SO₂ and O₃ in an open-air fumigation experiment. Sulphur dioxide treatment elevated foliar sulphur concentration in all species, and there were increases in foliar nitrogen in the two spruce species but not in pine. The concentrations of cations were frequently increased by SO₂ treatment, but there was no correlation between the sulphur concentration of needles and their total cation charge. SO₂-related elevations of foliar magnesium were correlated with the concentration of this element in soil solution, but the mechanism by which other cations were enhanced remains unclear. The only consistent effects on nutrient ratios were for SO₂ treatments to increase sulphur/cation ratios.     

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.     

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.     

Degradation of membrane lipid components and antiovidanl levels in Hordeum vulgare exposed to long-term fumigation with SO2

Hordeum vulgare L. (cv. Barberousse) seedlings were continuosly exposed to SO₂ (40±5 and 117±20 ppb) for 48 days and compared to control plants exposed to charcoal-filtered air. Glyco-and phospolipid amounts were strongly affected by exposure to SO_2:, and decreased with increasing SO₂, fumigation. The chloroplast membranes of fumigated leaves were characterized by lower quantities of rnonogalacto-syldiacylglycerol (MGDG) as well as of sulphoquinovosyldiacylglycerol (SQDG). Among the phospholipids a decrease in the phosphatidylglycerol (PG) content was observed, indicating that the chloroplast membranes are affected even by such low concentrations of SO₂ as used here. As for the other phospholipids, phosphatidylcholine (PC) and phosphatidylinositol (PI) suffered a reduction and phosphatidylethano-lairrine (PE) almost disappeared at the highest SO₂, concentration, whereas phosphatidic acid (PA) followed the opposite trend. The fatty acids of both glyco- and phospholipids fell sharply upon fumigation without any decline in the degree of unsataration. In particuSar, neither linoleic nor linolenic acid showed any evident change in level, suggesting that no lipid peroxidation of polytmsaturated fatty acid tails occurred. Furthermore, the electron spin resonance (ESR) showed no differences in the free radical levels between control and fumigated leaves. The level of malondialdehyde (MDA) decreased, while lipid-soluble antioxidants, as well as superoxide dismutase (SOD; EC 1.15.1.1) and catalase (EC 1.11.1.6) activities increased with the increase in SO₂ concentration, providing evidence that defence mechanisms against oxy-radical action are activated by the plants at low levels of SO₂,. Even if SO₂, at low concentrations may not directly oxidize polyunsaturated fatty acids, it may act as a deesterifying factor on each component of both glyco- and phospholipids.     

Effects of elevated CO2, elevated O3 and potassium deficiency on Norway spruce [Picea abies (L) Karst.]: seasonal changes in photosynthesis and non-structural carbohydrate content

Two clones of 5-year-old Norway spruce [Picea abies (L.) Karst.] were exposed to two atmospheric concentrations of CO2 (350 and 750 μmol mol^?1) and O₃ (20 and 75nmolmol^?1) in a phytotron at the GSF-Forschung-szentrum (Munich) over the course of a single season (April to October). The phytotron was programmed to recreate an artificial climate similar to that at a high elevation site in the Inner Bavarian Forest, and trees were grown in large containers of forest soil fertilized to achieve contrasting levels of potassium nutrition, designated well-fertilized or K-deficient. Measurements of the rate of net CO₂ assimilation were made on individual needle year age classes over the course of the season, chlorophyll fluorescence kinetics were recorded after approximately 23 weeks, and seasonal changes in non-structural carbohydrate composition of the current year's foliage were monitored. Ozone was found to have contrasting effects on the rate of net CO₂ assimilation in different needle age classes. After c. 5 months of fumigation, elevated O₃ increased (by 33%) the rate of photosynthesis in the current year's needles. However, O3 depressed (by 30%) the photo-synthetic rate of the previous year's needles throughout the period of exposure. Chlorophyll fluorescence measurements indicated that changes in photosystem II electron transport played no significant role in the effects of O₃ on photosynthesis. The reasons for the contrasting effects of O₃ on needles of different ages are discussed in the light of other recent findings. Although O₃ enhanced the rate at which CO₂ was fixed in the current year's foliage, this was not reflected in increases in the non-structural carbohydrate content of the needles. The transfer of ambient CO₂-grown trees to a CO₂-enriched atmosphere resulted in marked stimulation in the photosynthetic rate of current and previous year's foliage. However, following expansion of the current year's growth, the photosynthetic rate of the previous year's foliage declined. The extent of photosynthetic adjustment in response to prolonged exposure to elevated CO₂ depended upon the clone, providing evidence of intraspecific variation in the long-term response of photosynthesis to elevated CO₂. The increase in photosynthesis induced by CO₂ enrichment was associated with increased foliar concentrations of glucose, fructose and starch (but no change in sucrose) in the new growth. CO₂ enrichment significantly enhanced the photosynthetic rate of K-deficient needles, but there was a strong CO₂soil interaction in the current year's needles, indicating that the long-term response of trees to a high CO₂ environment may depend on soil fertility. Although the rate of photosynthesis and non-structural carbohydrate content of the new needles were increased in O₃-treated plants grown at higher levels of CO₂, there was no evidence that elevated CO₂ provided additional protection against O₃ damage. Simultaneous exposure to elevated O₃ modified the effects of elevated CO₂ on needle photosynthesis and non-structural carbohydrate content, emphasizing the need to take into account not only soil nutrient status but also the impact of concurrent increases in photochemical oxidant pollution in any serious consideration of the effects of climate change on plant production.     

RuBPCO kinetics and the mechanism of CO2 entry in C3 plants

Abstract. The CO₂ partial pressure in the chloroplasts of intact photosynthetic C₃ leaves is thought to be less than the intercellular CO₂ partial pressure. The intercellular CO₂ partial pressure can be calculated from CO₂ and H₂O gas exchange measurements, whereas the CO₂ partial pressure in the chloroplasts is unknown. The conductance of CO₂ from the intercellular space to the chloroplast stroma and the CO₂ partial pressure in the chloroplast stroma can be calculated if the properties of photosynthetic gas exchange are compared with the kinetics of the enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBPCO). A discrepancy between gas exchange and RuBPCO kinetics can be attributed to a deviation of CO₂ partial pressure in the chloroplast stroma from that calculated in the intercellular space. This paper is concerned with the following: estimation of the kinetic constants of RuBPCO and their comparison with the CO₂ compensation concentration; their comparison with differential uptake of ¹⁴CO₂ and ¹²CO₂; and their comparison with O₂ dependence of net CO₂ uptake of photosynthetic leaves. Discrepancy between RuBPCO kinetics and gas exchange was found at a temperature of 12.5 °C, a photosynthetic photon flux density (PPFD) of 550 μmol quanta m^?2 s^?1, and an ambient CO₂ partial pressure of 40 Pa. Consistency between RuBPCO kinetics and gas exchange was found if CO₂ partial pressure was decreased, temperature incresed and PPFD decreased. The results suggest that a discrepancy between RuBPCO kinetics and gas exchange is due to a diffusion resistance for CO₂ across the chloroplast envelope which decreases with increasing temperature. At low CO₂ partial pressure, the diffusion resistance appears to be counterbalanced by active CO₂ (or HCO₃) transport with high affinity and low maximum velocity. At low PPFD, CO₂ partial pressure in the chloroplast stroma appears to be in equilibrium with that in the intercellular space due to low CO₂ flux.     

Physiological effects of a long term exposure to low concentrations of NH3, NO2 and SO2 on Douglas fir (Pseudotsuga menziesii)

The above-ground parts of two years old seedlings of Douglas fir (Pseudotsuga menziesii) were exposed to filtered air, NH₃, NO₂₊, SO₂ (66, 96 and 95 μg m^?3, respectively), to a mixture of NO₂+NH₃ (55 + 82 μg m^?3) or SO₂+NO₂ (128 + 129 μg m^?3), for 8 months in fumigation chambers. Both chlorophyll fluorescence and gas exchange measurements were carried out on shoots which had sprouted at the beginning of the exposure period. The chlorophyll fluorescence measurements were performed after 3 and 5 months of exposure (average shoot age 70 and 140 days, respectively). Light response curves of electron transport rate (J) were determined, in which J was deduced from chlorophyll fluorescence. In addition, light response curves of net CO₂ assimilation were determined after 5 months of exposure. After 3 months of exposure (average shoot age 70 days) all exposure treatments showed a lower maximum electron transport rate (J_max) as compared to the control shoots (filtered air). A large reduction (45%) was observed for shoots exposed to SO₂+NO₂. During the exposure period between 3 and 5 months (average shoot age 70 and 140 days, respectively) a decrease of J_max was observed for all treatments. J_max had further declined some time after termination of the exposure, when average shoot age was 310 days. Shoots exposed to SO₂ and SO₂+NO₂ also showed a reduction in maximum net CO₂ assimilation (P_max) as compared to the control shoots. However, shoots exposed to NO₂ showed no reduction and even a higher P_max was observed for shoots exposed to NH₃ or NO₂+NH₃. Needles of these treatments also showed a higher chlorophyll content which might explain the contradictory results obtained for these treatments: the increased amount of photosynthetic units counteracts the reduction in J_max and consequently no reduction in P_max is measured. Shoots exposed to SO₂ and SO₂+NO₂ also showed a reduction in maximum stomatal conductance (g_s). However, the stomatal opening was larger than could be expected on basis of their (maximum) CO₂ assimilation rate. Consequently, water use efficiency of these shoots was lower than that of the control shoots. Also shoots exposed to NO₂ had a lower water use efficiency due to a significantly higher maximum g_s. Shoots exposed to NH₃ showed a high transpiration rate in the dark, indicating imperfect stomatal closure.     

The effects of SO2 and O3 fumigation on acid deposition and foliar leaching in the Liphook forest fumigation experiment

The ambient pollution climate at the Liphook forest fumigation site, where coniferous trees were fumigated with SO₂ and O₃, for 4 years under field conditions, was characteristic of the fringes of the areas where pollutant effects are a problem. Experimental treatments increased SO₂ concentrations to levels more characteristic of Eastern Europe, and summer O₃ concentrations by 30%. Deposition of SO₂ to the soil between the trees (inferred from shallow lysimeters) was significant, the deposition velocity being 2–1 mms^?1. Deposition to Scots pine and Sitka spruce canopies was greater, deposition velocities being 8.5 and 9.4 mm s^?1, respectively. These high values may perhaps be explained by co-deposition with NH₃. Calculations assume that dry deposition was the sole source of SO₄^2? gain in throughfall, and that there was no significant retention by the trees. There was a trend for O₃ to enhance SO₂ deposition to both soil and trees. Fumigation with SO₂ led to a significant increase in leaching of cations from foliage. Each species neutralized about 63% of the dry-deposited SO₂, predominantly by ion exchange for Ca and K. Equations are provided which allow calculation of foliar leaching given SO₂ concentrations or SO₄^2? deposition. Fumigation increased the rate of nutrient cycling considerably, without affecting foliar concentrations or damaging the trees. Ozone treatments did not enhance foliar leaching, calling into question some suggested mechanisms for the causes of forest decline.     

A branch bag and CO2 control system for long-term CO2 enrichment of mature Sitka spruce [Picea sitchensis (Bong.) Carr.]

This paper describes the construction and performance of branch bags and a CO₂ control system used to fumigate branches of mature Sitka spruce trees with air enriched in CO₂ (700 μmolmol^-1). It contains some examples of results obtained using the system over the course of the first two growing seasons. The branch bags have run continuously for 2 years with very few problems. CO₂ concentrations were within 20 μmol mol^-1 of the target concentration for more than 90% of the time. Temperatures within the bags were slightly higher than ambient (1–2 °C) and this had some effect on phenology. Attenuation of quantum flux density (photosynthetically active radiation) was 10–15%. The branch bag system has enabled investigation into the effects of elevated CO₂ on mature tissue without the problems and expense of fumigating whole trees. Growth in elevated CO₂ resulted in an increase in starch and a decrease in soluble protein content of needles. Stomatal conductance was higher in elevated CO₂ grown needles, and there was some evidence of an increase in photosynthetic capacity.     

Photosynthetic capacity,chloroplast pigments,and mineral content of the previous year's spruce needles with and without the new flush: analysis of the forest-decline phenomenon of needle bleaching

Summary Spruce (Picea abies) damage in the Fichtelgebirge (FRG) occurs as needle bleaching and a depression of CO₂ assimilation. Such injury may primarily result from the direct, above-ground effects of air pollution or indirect, below-ground changes in mineral uptake.Typically, the new flush of spruce needles is green and exhibits high photosynthetic capacity. Mies and Zöttl concluded that the older foliage is damaged when nutrients are withdrawn to supply the current year's needles. By removing the terminal buds of single branches in the spring, we produced an experimental set of the previous year's needles with greater mineral reserves than the control needles. During the course of the growing period, the performance of the experimental needles, which lacked competition from the new flush, was compared to that of the control needles of the same age-class on intact branches with the new flush.Throughout the experiment, chloroplast pigments of a healthy control tree were not affected by the elimination of the new flush. However, the chlorophyll and carotenoid content as well as the photosynthetic capacity of the previous year's needles on those branches of a heavily damaged tree where the new flush had been eliminated increased substantially. This increase was associated with an increase in minerals, which seemed to be deficient in the control needles with the new flush. Thus, in contrast to needles of the same age-class on intact branches with undisturbed new growth in the same atmospheric environment, the experimental needles escaped bleaching and a decrease in photosynthesis. It would seem that the bleaching and the loss in photosynthetic capacity typical of trees damaged by forest decline indirectly result from nutrient deficiencies through soil environment changes and/or root damage than directly from atmospheric pollutants.     

Plant responses to H2S and SO2 fumigation. I. Effects on growth, transpiration and sulfur content of spinach

Exposure of spinach (Spinacia oleracea L. cv. Monosa) to 0.25 μl l_?1 H₂S reduced the relative growth rate by 26, 47 and 60% at 15, 18 and 25°C, respectively. Shoot to root ratio decreased in plants fumigated at 18 and 25°C. Growth of spinach was not affected by a 2-week exposure to 0.10 or 0.25 μl l_?1 SO₂. Both H₂S and SO₂ fumigation increased the content of sulfhydryl compounds and sulfate. A 2-week exposure to 0.25 μl l_?1 H₂S resulted in an increase in sulfhydryl and sulfate content of 250 to 450% and 63 to 248% in the shoots, respectively, depending on growth temperature. Exposure to 0.15 and 0.30 μl l_?1 H₂S at 20°C for 2 weeks resulted in a 46% increase in sulfate content of the shoots at 0.30 μl l_?1 and no detectable increase at 0.15 μl l_?1 H₂S; the sulfate content of the roots increased by 195 and 145% at 0.15 and 0.30 μl l_?1 H₂S, respectively. Fumigation with 0.25 μl l_?1 SO₂ at 20°C for 2 weeks resulted in an increase in sulfhydryl content and sulfate content in the shoots of 285% and 300 to 1100%. H₂S fumigation during the 12 h light period or only during the dark period resulted in identical growth reduction and accumulation of sulfhydryl compounds; they were about 50 and 67% of those observed in continuously exposed plants. H₂S- and SO₂-exposed plants showed an increased transpiration rate, which was mainly caused by an increased dark-period transpiration. No effect of H₂S and SO₂ on the water uptake of the plants and the osmotic potential of the leaves was detected. Plants fumigated with 0.25 μl l_?1 H₂S for 2 weeks were smaller and differed morphologically from the control plants by slightly more abaxially curved leaf margins. Cross sections of the leaves showed smaller cells at the margins and smaller and fewer air spaces. The increased transpiration in the H₂S-exposed plants is discussed in relation to the observed morphological changes.     

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.     

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.     

Changes in Photosystem Ⅱ Activity and Leaf Reflectance Features of Several Subtropical Woody Plants Under Simulated SO2 Treatment

The effects of simulated SO2 treatment on the photosynthetic apparatus were investigated in five subtropical forest plants, namely Plnus massonlana Lamb., Schlma superba Gardn. et Champ., Castanopsls flssa （Champ. ex Benth.） Rehd. et Wils., Acmena acuminatissima （BI.） Merr et Perry, and Cryptocarya concinna Hance. After leaf sections had been immersed in 0, 20, 50, and 100 mmol/L NaHSO3 for 20 h, total chlorophyll （Chl） content, Chl a/b, maximal photochemical efficiency, and the photochemical quantum yields of photosystem Ⅱ of all five woody plants were reduced to different degrees, whereas lutein content （Chl base） was increased. Two protective mechanisms, namely the xanthophyll cycle （de-epoxidation） and an anti-oxidant system （1,1-diphenyl-2-picrylhydrazyl radical-scavenging capacity）, showed differences in the degree of modulation under simulated SO2 treatment. Compared with control （distilled water treatment）, the revised normalized difference vegetation index, a leaf reflectance index, was lowered with Increasing concentrations of NaHSO3. Cryptocarya conclnna, a dominant species in the late succession stage of subtropical forests in South China, exhibited less sensitivity to NaHSO3. Conversely, Pinus massonlana, the pioneer hellophyte species, was most susceptible to NaHSO3 treatment. It Is suggested that SO2 pollution may accelerate the succession of subtropical forest.     

Leaf surface properties of Norway spruce needles exposed to sulphur dioxide and ozone in an open-air fumigation system at Liphook

Samples of current-year and 1-year-old foliage were taken from Norway spruce (Picea abies (L.) Karst.) trees in April 1991, 4 months after a 3–4 year controlled fumigation with O₃ and SO₂ in the open at Liphook, south-east England. Trees were grown in seven plots, and treated in a factorial design with three levels of SO₂ and two levels of O₃ (ambient and c. 1.3 × ambient), with an extra ambient air plot. All statistical analyses were made on plot means. Leaf wettability, as measured by the contact angle of water droplets, was significantly affected by needle age and by SO₂ treatment (P≤0–05. in older needles, decreasing with increasing SO₂ concentration. There was no effect of O₃ on wettability, and no effect of any treatment on amounts of surface wax extracted by immersion of needles in chloroform. Electrolyte leakage rates from detached current-year needles were not affected by prior exposure to O₃, but decreased significantly (P= 0.034) with increasing exposure to SO₂. There was no detectable effect of fumigation on the rate of water loss from detached needles. Similarly, there was no effect of fumigation on the dry weight/fresh weight ratio of needles. The total sulphur content of needles increased significantly (P≤0.0001) with exposure to SO₂ and with needle age. Amounts of water-extractable sulphate, however, varied greatly among plots, but with no pattern with respect to fumigation treatment. It is concluded that leaf wettability and electrolyte leakage rates may be good indicators of the persistent effects of SO₂ on Norway spruce growing in the open air, and that the observed changes in leaf surface properties in response to SO₂ fumigation have implications for the processes, both biotic and abiotic, that occur on leaf surfaces.     

Photosynthetic CO2 fixation in spinach chloroplasts inhibited by pine chloroplasts or extracts of pine chloroplasts

Chloroplasts isolated from pine needles were found to be inactive with respect to CO₂ fixation. Since it was suspected that pine needles may contain substances inhibitory to photosynthesis, studies were carried out using photosynthetically active isolated spinach chloroplasts and chloroplasts isolated from pine needles. When isolated pine chloroplasts were suspended in buffer and were added to isolated spinach chloroplasts they inhibited photosynthetic CO₂ fixation. When the pine chloroplasts were separated from the medium by centrifugation, the separated pine chloroplasts severely inhibited CO₂ fixation by isolated spinach chloroplasts, but the supernatant solution from the pine chloroplasts was not inhibitory. As little as 5% pine chloroplasts (based on chlorophyll content) produced 50% inhibition of CO₂ fixation by the spinach chloroplasts. Studies of fixation of ¹⁴C-labelled CO₂ by spinach chloroplasts were carried out in which after 5 min photosynthesis the pine chloroplasts were added. It was found that the subsequent inhibition of spinach CO₂ fixation was neither due to any effect on the rate of export of photosynthetic metabolites from the chloroplasts to the medium, nor to a direct effect on the RUBP carboxylase reaction. The principal effect was found to be an inhibition of the conversion of fructose-1,6-bisphosphate and sedoheptulose-1,7-bisphosphate to the respective monophosphates and inorganic phosphate. From this finding it was concluded that a principal effect of the inhibition by pine chloroplasts is probably an inhibition either directly or indirectly of the bisphosphatase enzymes in the spinach chloroplasts. Based on its distribution between organic and aqueous acidic or neutral solutions, the inhibitory factor of the pine chloroplasts must be lipophilic. Most of the factor could be transferred to an aqueous phase in a strongly alkaline solution. Following subsequent acidification of the aqueous phase the activity could be completely transferred back into the organic phase. This procedure allowed for separation of the inhibitory factor from most of the pigments and other lipophilic substances present in the pine chloroplasts and yielded a preparation which could be subsequently fractionated by thin layer chromatography. UV absorption was found in two fast moving spots and at the origin. The fastest running spot from the thin layer chromatography plate was found to be the one containing most of the inhibitory activity.     

Effects of elevated O3 and CO2 concentrations on photosynthesis and stomatal conductance in Scots pine

Naturally regenerated Scots pines (Pinus sylvestris L.), aged 28–30 years old, were grown in open-top chambers and subjected in situ to three ozone (O₃) regimes, two concentrations of CO₂, and a combination of O₃ and CO₂ treatments From 15 April to 15 September for two growing seasons (1994 and 1995). The gas exchanges of current-year and 1-year-old shoots were measured, along with the nitrogen content of needles. In order to investigate the factors underlying modifications in photosynthesis, five parameters linked to photosynthetic performance and three to stomatal conductance were determined. Elevated O₃ concentrations led to a significant decline in the CO₂ compensation point (Г^*), maximum RuP₂-saturated rate of carboxylation (V_cmax), maximum rate of electron transport (J_max), maximum stomatal conductance (g_smax), and sensitivity of stomatal conductance to changes in leaf-to-air vapour pressure difference (?g_s/?D_v) in both shoot-age classes. However, the effect of elevated O₃ concentrations on the respiration rate in light (R_d) was dependent on shoot age. Elevated CO₂(700 μmol mol^?1) significantly decreased J_max and g_smax but increased R_d in 1-year-old shoots and the ?g_s/?D_v in both shoot-age classes. The interactive effects of O₃ and CO₂ on some key parameters (e.g. V_cmax and J_max) were significant. This may be closely related to regulation of the maximum stomatal conductance and stomatal sensitivity induced by elevated CO₂. As a consequence, the injury induced by O₃ was reduced through decreased ozone uptake in 1-year-old shoots, but not in the current-year shoots. Compared to ambient O₃ concentration, reduced O₃ concentrations (charcoal-filtered air) did not lead to significant changes in any of the measured parameters. Compared to the control treatment, calculations showed that elevated O₃ concentrations decreased the apparent quantum yield by 15% and by 18%, and the maximum rate of photosynthesis by 21% and by 29% in the current-year and 1-year-old shoots, respectively. Changes in the nitrogen content of needles resulting from the various treatments were associated with modifications in photosynthetic components.