Diagnostic parameters for selecting against novel spruce (Picea abies) decline: IV. Response of photosynthesis and transpiration to SO2+NO2 exposures

The dose- and time-response effects of 3 days of 6 h day-time sequential exposures to NO₂, SO₂ and SO₂+NO₂ of 0.45–1.81 μl l⁻¹ (ppm) SO₂ and 1.50–7.65 μl l⁻¹ NO₂ on photosynthesis, transpiration and dark respiration were examined for nine Carpatho-Ukrainian half-sib families and a population from the GFR ('Westerhof') of Norway spruce [ Piecea abies (L.) Karst.], all in their 5th growing season.
SO₂+NO₂ inhibited photosynthesis and transpiration and stimulated dark respiration more than SO₂ alone. SO₂ and SO₂+NO₂ at the lowest concentrations inhibited night transpiration, but increased it at the highest concentration, the strongest effects being obtained with combined exposures. Photosynthesis of the different half-sib families was affected significantly differently by SO₂+NO₂ exposures. NO₂ alone had no effects.
Sensitivity to transpiration decline correlated negatively with branch density. Height of trees correlated postitively with decline sensitivity in the seed orchard. The distribution of photosynthesis and transpiration sensitivities over all tested half-sib families correlated negatively with the distribution of decline sensitivity of their parents in a rural Danish seed orchard. The relative photosynthesis and transpiration sensitivities may thus serve as diagnostic parameters for selecting against novel spruce decline.     

CO2 enrichment and development of freezing tolerance in Norway spruce

Plant growth and adaptation to cold and freezing temperatures in a CO₂-enriched atmosphere have received little attention despite their predicted effects on plant distribution and productivity. In this study we looked at the interaction between elevated CO₂ and development of freezing tolerance in Norway spruce ( Picea abies (L.) Karst.). First-year seedlings were grown under controlled conditions in an atmosphere enriched in CO₂ (70 Pa) for one simulated growth season. We measured shoot growth, registered the timing of growth cessation and bud set, measured needle net photosynthetic rate, and determined needle carbohydrate concentration (fructose+pinitol, glucose, sucrose, inositol, raffinose and starch). Freezing tolerance (LT₅₀) was determined after exposing whole seedlings to temperatures ranging from −6.5 to −36.0°C and scoring for visual needle browning. Elevated CO₂ did not affect height growth or the timing of growth cessation and bud set. The only statistically significant effects of CO₂ treatment were on seedling dry weight, percent dry matter and starch content. During the three weeks after growth cessation and bud set, freezing tolerance increased from −10 to −35°C, and there was a marked increase in all soluble sugars except inositol. However, neither freezing tolerance nor the concentration of soluble sugars was significantly influenced by elevated CO₂.     

Free fatty acids, neutral and polar lipids in Hordeum vulgare exposed to long-term fumigation with SO2

Three 9-day-old cultivars of Hordeum vulgare L. (Barberousse, Gerbel and Panda) were exposed to low levels of SO₂ fumigation (40 ± 5 and 117 ± 20 ppb). After 48 days of treatment the seedlings were harvested for lipid analysis. In comparison to the control (plants exposed to charcoal-filtered air), the total lipid content of fumigated seedlings declined at 40 ppb SO₂ and even more so at 117 ppb in all three cultivars. A large reduction in diacylglycerols. polar lipids and free sterols was also observed after fumigations at both SO₂ concentrations, whereas the treatments resulted in an increase in triacylglycerols and free fatty acids. The percentage composition of total fatty acids and that of each lipid class were changed by the fumigations. resulting in an increase in the degree of unsaturation. No changes in the percentage composition of sterols occurred in the fumigated leaves. These results suggest that even if SO₂ may not directly oxidize unsaturated fatty acids at the low concentrations used here (which do not cause visible injury). it may alter lipid metabolism. This alteration. which was particularly evident in the polar lipids and sterols, could affect the functions associated with membrane stabilization, in which lipids plus sterols play a key part.     

Plant responses to H2S and SO2 fumigation. II. Differences in metabolism of H2S and SO2 in spinach

Fumigation of spinach (Spinacia oleracea L. cvs Estivato and Monosa) with H₂S or SO, for 1 to 6 days resulted in accumulation of sulfhydryl (SH) compounds in the shoots of both H₂S- and SO₂-exposed plants. The sulfate concentration in shoots of SO₂-exposed plants increased linearly with time. SH accumulation showed saturation kinetics as a function of time as well as H₂S concentration, ascribed to the internal H₂S concentration in the plant and the availability of substrates for glutathione synthesis, respectively. SH compounds accumulated more at lower exposure temperatures, whereas sulfate accumulation was more pronounced at higher temperatures. These results are discussed in relation to the possible foliar uptake of H₂S and SO₂, the temperature dependence of uptake and the water solubility of these gases. The possibility of SO₂-induced H₂S emission rather than sulfate accumulation as a source for SH accumulation is also discussed. Cessation of fumigation resulted in a decrease in SH compounds and sulfate content that could be accounted for by sulfur metabolism and growth, respectively.     

Gas exchange and carbon isotope composition of Ananas comosus in response to elevated CO2 and temperature

Ananas comosus L. (Merr.) (pineapple) was grown at three day/night temperatures and 350 (ambient) and 700 (elevated) μ mol mol^–1 CO₂ to examine the interactive effects of these factors on leaf gas exchange and stable carbon isotope discrimination ( Δ ,‰). All data were collected on the youngest mature leaf for 24 h every 6 weeks. CO₂ uptake (mmol m^–2 d^–1) at ambient and elevated CO₂, respectively, were 306 and 352 at 30/20 °C, 175 and 346 at 30/25 °C and 187 and 343 at 35/25 °C. CO₂ enrichment enhanced CO₂ uptake substantially in the day in all environments. Uptake at night at elevated CO₂, relative to that at ambient CO₂, was unchanged at 30/20 °C, but was 80% higher at 30/25 °C and 44% higher at 35/25 °C suggesting that phosphoenolpyruvate carboxylase was not CO₂-saturated at ambient CO₂ levels and a 25 °C night temperature. Photosynthetic water use efficiency (WUE) was higher at elevated than at ambient CO₂. Leaf Δ -values were higher at elevated than at ambient CO₂ due to relatively higher assimilation in the light. Leaf Δ was significantly and linearly related to the fraction of total CO₂ assimilated at night. The data suggest that a simultaneous increase in CO₂ level and temperature associated with global warming would enhance carbon assimilation, increase WUE, and reduce the temperature dependence of CO₂ uptake by A. comosus .     

The role of temperature in determining the stimulation of CO2 assimilation at elevated carbon dioxide concentration in soybean seedlings

Soybean ( Glycine max cv. Clark) was grown at both ambient (ca 350 μmol mol⁻¹) and elevated (ca 700 μmol mol⁻¹) CO₂ concentration at 5 growth temperatures (constant day/night temperatures of 20, 25, 30, 35 and 40°C) for 17–22 days after sowing to determine the interaction between temperature and CO₂ concentration on photosynthesis (measured as A, the rate of CO₂ assimilation per unit leaf area) at both the single leaf and whole plant level. Single leaves of soybean demonstrated increasingly greater stimulation of A at elevated CO₂ as temperature increased from 25 to 35°C (i.e. optimal growth rates). At 40°C, primary leaves failed to develop and plants eventually died. In contrast, for both whole plant A and total biomass production, increasing temperature resulted in less stimulation by elevated CO₂ concentration. For whole plants, increased CO₂ stimulated leaf area more as growth temperature increased. Differences between the response of A to elevated CO₂ for single leaves and whole plants may be related to increased self-shading experienced by whole plants at elevated CO₂ as temperature increased. Results from the present study suggest that self-shading could limit the response of CO₂ assimilation rate and the growth response of soybean plants if temperature and CO₂ increase concurrently, and illustrate that light may be an important consideration in predicting the relative stimulation of photosynthesis by elevated CO₂ at the whole plant level.     

SO2 and assimilatory sulfate reduction in beech leaves

The effect of SO₂ on the extractable activity of ATP sulfurylase (EC 2.7.7.4.). adenosine 5'-phosphosulfate sulfotransferase, ribulosebisphosphate carboxylase, chlorophyll, protein, sulfate, and amino acids was examined in leaves of potted grafts of beech ( Fagus sylvatica L.) treated in outdoor fumigation chambers. Addition of 0.025 and 0.075 μl SO₂ 1⁻¹ to unfiltered ambient air caused a decrease in the extractable activity of adenosine 5'-phosphosulfate sulfotransferase to about 20 to 30% of the controls. Neither the extractable activity of ATP sulfurylase and ribulosebisphosphate carboxylase nor the content in chlorophyll, total amino acids and protein were significantly affected by SO₂, but there was an increase in the sulfate content. Leaves treated with 0.075 μl SO₂ 1⁻¹ contained more alanine and cysteine and less serine than the controls. After transfer of the SO2-treated beech trees to control chambers there was an increase in adenosine 5'-phosphosulfate sulfotransferase activity, but no significant decrease in SO₂₋⁴-sulfur.     

Effects of raised CO2 on potential CH4 production and oxidation in, and CH4 emission from, a boreal mire

1 In a glasshouse experiment we studied the effect of raised CO₂ concentration (720 p.p.m.) on CH₄ emission at natural boreal peat temperatures using intact cores of boreal peat with living vascular plants and Sphagnum mosses. After the end of the growing season half of the cores were kept unnaturally warm (17–20 °C). The potential for CH₄ production and oxidation was measured at the end of the emission experiment.
2 The vascular cores ('Sedge') consisted of a moss layer with sedges, and the moss cores (' Sphagnum ') of Sphagnum mosses (some sedge seedlings were removed by cutting). Methane efflux was 6–12 times higher from the Sedge cores than from the Sphagnum cores. The release of CH ₄ from Sedge cores increased with increasing temperature of the peat and decreased with decreasing temperature. Methane efflux from Sphagnum cores was quite stable independent of the peat temperatures.
3 In both Sedge and Sphagnum samples, CO₂ treatment doubled the potential CH₄ production but had no effect on the potential CH₄ oxidation. A raised concentration of CO₂ increased CH₄ efflux weakly and only at the highest peat temperatures (17–20 °C).
4 The results suggest that in cool regions, such as boreal wetlands, temperature would restrict decomposition of the extra substrates probably derived from enhanced primary production of mire vegetation under raised CO₂ concentrations, and would thus retard any consequent increase in CH₄ emission.     

Induction of nitrate reductase in needles of Scots pine seedlings by NOX and NO3−

The possibility to induce nitrate reductase (NR; EC 1.6.6.2) in needles of Scots pine ( Pinus sylvestris L.) seedlings was studied. The NR activity was measured by an in vivo assay. Although increased NR activities were found in the roots after application of NO₃⁻, no such increase could be detected in the needles. Detached seedlings placed in NO₃⁻ solution showed increasing NR activities with increasing NO₃⁻ concentrations. Exposure of seedlings to NO_x (70–80 ppb NO₂ and 8–12ppb NO) resulted in an increase of the NR activity from 10–20 nmol NO₂⁻ (g fresh weight)⁻¹ h⁻¹ to about 400 nmol NO₂⁻ (g fresh weight)⁻¹ h⁻¹. This level was reached after 2–4 days of exposure, thereafter the NR activity decreased to about 200 nmol NO₂⁻ (g fresh weight)⁻¹ h⁻¹. Analyses of free amino acids showed low concentrations of arginine and glutamine in NO_x-fumigated seedlings compared to corresponding controls.     

The effect of nitrogen source on photosynthesis of carob at high CO2 concentrations

Carob seedlings ( Ceratonia siliqua L. cv. Mulata), fed with nitrate or ammonium, were grown in growth chambers containing two levels of CO₂ (360 or 800 μl l⁻¹), three root temperatures (15, 20 or 25°C), and the same shoot temperature (20/24°C, night/day temperature). The response of the plants to CO₂ enrichment was affected by environmental factors such as the type of inorganic nitrogen in the medium and root temperature. Increasing root temperature enhanced photosynthesis rate more in the presence of nitrate than in the presence of ammonium. Differences in photosynthetic products were also observed between nitrate- and ammonium-fed carob seedlings. Nitrate-grown plants showed an enhanced content of sucrose, while ammonium led to enhanced storage of starch. Increase in root temperature caused an increase in dry mass of the plants of similar proportions in both nitrogen sources. The enhancement of the rates of photosynthesis by CO₂ enrichment was proportionally much larger than the resulting increases in dry mass production when nitrate was the nitrogen source. Ammonium was the preferred nitrogen source for carob at both ambient and high CO₂ concentrations. The level of photosynthesis of a plant is limited not only by atmospheric CO₂ concentration but also by the nutritional and environmental conditions of the root.     

Growth and photosynthetic response of three soybean cultivars to simultaneous increases in growth temperature and CO2

Three soybean ( Glycine max L. Merr.) cultivars (Maple Glen, Clark and CNS) were exposed to three CO₂ concentrations (370, 555 and 740 μmol mol⁻¹) and three growth temperatures (20/15°, 25/20° and 31/26°C, day/night) to determine intraspecific differences in single leaf/whole plant photosynthesis, growth and partitioning, phenology and final biomass. Based on known carboxylation kinetics, a synergistic effect between temperature and CO₂ on growth and photosynthesis was predicted since elevated CO₂ increases photosynthesis by reducing photorespiration and photorespiration increases with temperature. Increasing CO₂ concentrations resulted in a stimulation of single leaf photosynthesis for 40–60 days after emergence (DAE) at 20/15°C in all cultivars and for Maple Glen and CNS at all temperatures. For Clark, however, the onset of flowering at warmer temperatures coincided with the loss of stimulation in single leaf photosynthesis at elevated CO₂ concentrations. Despite the season-long stimulation of single leaf photosynthesis, elevated CO₂ concentrations did not increase whole plant photosynthesis except at the highest growth temperature in Maple Glen and CNS, and there was no synergistic effect on final biomass. Instead, the stimulatory effect of CO₂ on growth was delayed by higher temperatures. Data from this experiment suggest that: (1) intraspecific variation could be used to select for optimum soybean cultivars with future climate change; and (2) the relationship between temperature and CO₂ concentration may be expressed differently at the leaf and whole plant levels and may not solely reflect known changes in carboxylation kinetics.     

Elevated atmospheric [CO2] promotes frost damage in evergreen tree seedlings

Growth under elevated [CO₂] promoted spring frost damage in field grown seedlings of snow gum ( Eucalyptus pauciflora Sieb. ex Spreng.), one of the most frost tolerant of eucalypts. Freezing began in the leaf midvein, consistent with it being a major site of frost damage under field conditions. The average ice nucleation temperature was higher in leaves grown under elevated [CO₂] (– 5·7 °C versus – 4·3 °C), consistent with the greater incidence of frost damage in these leaves (34% versus 68% of leaves damaged). These results have major implications for agriculture, forestry and vegetation dynamics, as an increase in frost susceptibility may reduce potential gains in productivity from CO₂ fertilization and may affect predictions of vegetation change based on increasing temperature.     

Effects of sulphur dioxide on leaf photosynthesis: the role of temperature and humidity

The effect of temperature and humidity on SO₂–induced photosynthetic depression was determined in gas exchange experiments with leaves of Vicia faba , L. Stomatal behaviour was sensitive to humidity resulting in higher uptake rates of SO₂ and stronger reductions of photosynthesis at low VPD (vapour pressure deficit). After a fumigation period of 2 h, when the photosynthetic rate had stabilized, photosynthesis of leaves exposed to SO₂ at 8°C was reduced much more than at 18°C at the same rate of SO₂ uptake. Data analysis with a mechanistic model revealed that this effect was due to the slower rate of S(IV) oxidation at lower temperatures, resulting in higher accumulation of S(IV) and thus stronger reduction of photosynthesis. These results were confirmed by experimental analyses of the S(IV) concentration in leaves following fumigation, which showed that more S(IV) accumulated in leaves exposed at a lower temperature. This may explain the high sensitivity of plants exposed to SO₂ under winter conditions, when both VPD and temperature are low.     

The effects of temperature and season on the O2 consumption of third-instar larvae of the goldenrod gall fly (Eurosta solidaginis)

Abstract. Third-instar larvae of the goldenrod gall fly ( Eurosta solidaginis Fitch) live inside ball galls on goldenrod plants from summer to the following spring.Because galls are highly exposed to the weather, larvae experience substantial variations in body temperature.This study documents the oxygen consumption of gall fly larvae with regard to the effects of ambient temperature, seasonal conditioning, and prior exposure to subzero temperature.The body mass of larvae doubles between the late summer and the autumn; it subsequently undergoes a modest decline by early winter.The O₂, consumption of field-acclimatized larvae increases with ambient temperature, especially between 0 and 10°C (Q₁₀= 2.6-3.4).The thermal sensitivity of metabolism declines at higher ambient temperatures, most notably during the autumn/early winter.After exposure to 15°C for 1 week, autumn and early winter larvae maintain much lower rates of O₂ consumption than do late summer specimens.Prior exposure to -5°C for 24 h did not influence the O₂ consumption of larvae.Low thermal sensitivity of O₂ consumption, especially at higher ambient temperatures, is an energy-sparing mechanism during seasonal inactivity.Indeed, the persistence of this metabolic pattern in larvae exposed to 15°C suggests that they have entered a state of diapause.     

Effects of atmospheric SO2 on Azolla and Anabaena symbiosis

The water fern Azolla pinnata R. Br. was fumigated for 1 week with either 25, 50 or 100 nl 1⁻¹ SO₂. The symbiosis of Azolla with Anabaena azollae (spp.) was severely damaged by atmospheric SO₂ even at concentrations as low as 25 nl 1⁻¹, with significant reductions in growth, reduction of C₂H₂, NH₃ assimilation, protein synthesis, and heterocyst development. These disturbances appear to be mainly responsible for the extreme sensitivity of this fern to atmospheric SO₂. Changes in violaxanthin/antheraxanthin and epoxy-lutein/lutein ratios also indicate that free radical products are induced by atmospheric SO₂. These results suggest that the Azolla-Anabaena symbiotic system is a very responsive and reliable lower plant model to study the detailed effects of total sulphur deposition upon the balances between various important plant metabolic processes.     

Chloroplast ultrastructure in two Crassulacean species and an F1 hybrid with differing biomass δ13C values

Abstract. The chloroplasts of two species of the Crassulaceae and their F₁ hybrid were compared by electron microscopy. The two species had contrasting leaf tissue δ¹³C values of −25°/_∞ ( Sedum greggii ) and −13°/_∞ ( Cremnophila linguifolia ), and the F₁ hybrid had a value of − 18°/_∞. S. greggii had a mean of 8.9 thylakoids per granum in contrast to C. linguifolia which had a mean of only 3.8 thylakoids per granum. The F₁ hybrid had a mean of 6.4 thylakoids per granum. Crystaloids were observed in S. greggii and the hybrid but not in C. linguifolia     

CATABOLISM OF PROSTAGLANDIN ENDOPEROXIDES INTO PROSTAGLANDIN E2 AND F2x BY THE RAT BRAIN

Abstract— Tritium labeled prostaglandin (PG) endoperoxides PGG₂ and PGH₂ were rapidly transformed (2 min, 37°C) in good yield (> 50%) by homogenates of whole rat brain into a mixture of products including prostaglandin E₂ and F_2x: under similar conditions (10min. 37°C) tritium labeled arachidonic acid remained essentially unoxidised. The ratio of PGE-like products: PGF_2x formed was approx 0.5 as determined by radio thin layer chromatography. This ratio changed to unity when homogenates of cerebral cortex or cerebral hemispheres were employed. On the other hand cerebellar homogenates formed PGF_2x in much greater amounts. The structures of the products were confirmed by mass spectrometry and were further supported by experiments using octadeuterio-endoperoxides. In the latter experiments the resulting PGE₂ and PGF_2x contained the expected seven and eight deuterium atoms respectively. Evidence for the formation of heptadeuterio PGD₂. heptadeuterio-6-keto-PGF₁, and hexadeuterio 12-hydroxyheptadecatrienoic acid was also obtained by mass spectrometry. These experiments demonstrate for the first time in brain tissue the biosynthesis of labeled prostaglandins from exogenous tritiated and deuterated precursors.     

Improvement of the 15N dilution method for estimation of absorption of NOx by plants supplied with 15N-labelled fertilizer

To develop further the methods for estimation of NOx absorption by plants supplied with ¹⁵N-labelled fertilizer, we proposed a new calculation method, total N fixed method (TNF), and compared with the ¹⁵N dilution method and the classical mass balance method (MB).
Hydroponically grown soybean plants were supplied with ¹⁵N-labelled nitrate and exposed to 200–250 nl l⁻¹ NO₂ for 7 d. The proportions of the N derived from NO₂ to total N in exposed plants were estimated by the three methods.
The reported rates of NO₂ absorption by several plant species, estimated by the ¹⁵N dilution method, were recalculated using the TNF method. The results of the two methods were compared and showed that: (1) The ¹⁵N dilution method overestimated the content of NO₂-N in exposed plants compared with the MB method whilst the TNF method produced estimations of NO₂-N closer to those by the MB method when the plants were supplied with 5 m M nitrate. (2) The differences in estimations between the MB method and either the ¹⁵N dilution method or the TNF method increased with decreasing supply of ¹⁵N-labelled nitrate to roots.     

Growth and solute leakage in Hordeum vulgaris exposed to long-term fumigation with low concentrations of SO2

Plants of three commercial cultivars of Hordeum vulgaris L. (Barberousse, Gerbel, Panda) were continuously exposed to two concentrations of SO₂ (40±5 and 117±20 ppb) against a control (charcoal-filtered air). Experiments were performed in fumigation chambers. Fumigation was started 9 days after seeding and plant material was harvested after 48 days. In none of the three cultivars did visible symptoms of injury appear on the leaves after exposure to SO₂, while they all showed a reduction in plant height and dry matter of shoots and roots and a significant increase in the shoot/root dry weight ratios depending on the cultivars. The time course of conductivity, and K⁺ Ca²⁺ and sugar effluxes were determined for 24 h. Leaves from cvs Gerbel and Panda exposed to SO₂ exhibited a progressive increase in conductivity, and in Ca²⁺ and sugar efflux (expressed as percentages of their total effluxes). Plusmn; efflux of the three cultivars and conductivity in cv. Barberousse increased only after 24 h incubation. A relative leakage ratio (RLR) was calculated from the UV absorption at 280 nm after 24 h incubation of the leaf strips and the total 280 nm UV absorption, obtained after liquid N₂ killing of the tissues. The RLR was well correlated with the total amino acids and sugars found in the leachate and can be used as a test for determining the tissue response to SO₂.     

The combination effect of pH, SO2, ethanol and temperature on the growth of Leuconostoc oenos

A factorial design was used to determine the main effects and interactions of pH (3.6, 3.8 and 4.0), SO₂ (25 and 50 mg/l), ethanol (7%, 10% and 13%, v/v) and temperature (15°C and 25°C) on the growth of 54 Leuconostoc oenos strains. These differed greatly in their ability to survive adverse conditions. Under the test conditions, temperature had a profound effect on growth. The power pH values of 3.6 and 3.8 had a marked inhibitory effect on growth, as did at the lower temperature of 15°C. The higher concentrations of SO₂ (50 mg/l) and ethanol (13%) had the greatest inhibitory effect on growth, and with a synergistic effect when both SO₂ and ethanol were added to the test medium. Based on these results, five strains were selected that were best capable of surviving the adverse conditions studied. These strains will be used for further study on the induction of malolactic fermentation in wine.