Sulfide-induced dissimilatory nitrate reduction to ammonia in anaerobic freshwater sediments

Abstract: Different reduced sulfur compounds (H₂S, FeS, S₂O₃²⁻) were tested as electron donors for dissimilatory nitrate reduction in nitrate-amended sediment slurries. Only in the free sulfide-enriched slurries was nitrate appreciably reduced to ammonia (     ), with concomitant oxidation of sulfide to S⁰ (     ). The initial concentration of free sulfide appears as a factor determining the type of nitrate reduction. At extremely low concentrations of free S²⁻ (metal sulfides) nitrate was reduced via denitrification whereas at higher S²⁻ concentrations, dissimilatory nitrate reduction to ammonia (DNRA) and incomplete denitrification to gaseous nitrogen oxides took place. Sulfide inhibition of NO- and N₂O- reductases is proposed as being responsible for the driving part of the electron flow from S²⁻ to NH₄⁺.     

The response of Norway spruce seedlings to simulated acid mist

Four pot experiments are reported in which Norway spruce ( Picea abies (L.) Karst) seedlings, of different nutrient status, were treated with acid mist for one growing season in open-top chambers (OTCs). Combinations of H⁺, SO₄²⁻, NH₄⁺ and NO₃⁻ were applied at different frequencies of application and supplying different doses of S and N kg ha⁻¹. Plant growth, visible injury, frost hardiness and nutrient status were observed. These experiments were undertaken to improve our understanding of the interaction of environmental factors such as nutrition and mist-exposure frequency on seedling response to N and S deposition.
Both acidity (pH 2·7) and SO₄²⁻ ions were necessary to induce visible injury. Mist containing SO₄²⁻, H⁺ and to a lesser extent NH₄⁺ significantly reduced winter frost hardiness. Increasing the misting frequency, and to a lesser extent the overall dose, increased the likelihood of acid mist causing visible injury and reducing frost hardiness. Post-planting stress, low N status and needle juvenility increased the likelihood of acid mist causing visible injury. Increased plant vitality, adequate N status and growth rate reduced the likelihood of acid-mist-induced reductions in frost hardiness.
Principles underlying the responses of spruce seedlings treated in controlled conditions to acid mist are discussed.     

Comparison of ion transportation before and after egg hatching in Amphinemura sp. (Plecoptera)

Abstract.  An increase in egg size with embryonic development in stoneflies is believed to result from the uptake of water by osmosis. The present study aims to investigate whether a selective ion transport through egg membranes exists before hatching, and whether ions are released after hatching. Viable and nonviable egg masses are incubated in Petri dishes filled with water, and the concentrations of the ions F⁻, Cl⁻, SO₄²⁻, NO₃⁻, Na⁺, K⁻, Mg²⁺ and Ca²⁺ in the water are determined. The ion transport of an egg mass before and after hatching and a nonviable egg mass is then calculated. Before hatching, Cl⁻, SO₄²⁻, NO₃⁻, Na⁺, Mg²⁺ and Ca²⁺ are taken up from the surrounding water into the inner egg. These ions are selectively taken into the egg. After hatching, Cl⁻, SO₄²⁻, Na⁺, Mg²⁺ and Ca²⁺ are released into the surrounding water. The amount of these ions released after hatching is lower than the amount taken up before hatching. Ions that are not released after hatching are considered to be used in embryonic development.     

A rapid and sensitive ion chromatographic technique for the determination of sulfate and sulfate reduction rates in freshwater lake sediments

Abstract Newly developed low capacity columns were used in suppressed ion chromatography for rapid and highly reproducible determination of SO₄²⁻ in porewater samples from freshwater sediments without preconcentration of samples. With a 50 μl injection the detection limit for SO₄²⁻ was ca. 50 pmol (= 1 μ M) with a precision of 1–3% at the 10–200 μM level and <1% at concentrations above 200 μM. SO₄²⁻ could be measured in 4–5 min with the routinely used eluent (3.0 mM NaHCO₃/0.8 mM Na₂CO₃). When the strength of the eluent was increased to 3.0 mM NaHCO₃/2.0 mM Na₂CO₃, sulfate analysis was possible in less than 3 min, provided that samples were nitrate-free. Under these conditions S₂O₃²⁻ could also be sensitively determined in about 6 min. Examples of application of the method are given for measurements of sulfate reduction rates in freshwater sediment samples from Lake Constance.     

The role of sulphur compounds for breeding success of Ips typographus L. (Col., Scolytidae) on Norway Spruce (Picea abies [L.] Karst.)

Bark beetles, especially Ips typographus L. represent a severe biotic threat for spruce ( Picea abies [L.] Karst.) at low altitudes in Europe. We compared sulphur (total S, SO₄²⁻, glutathione, cysteine, methionine), nitrogen (total N, NO₃⁻, total protein, free amino acids), carbon, total phosphorus and PO₄³⁻, tree vigour index (TVI) and water content of the phloem after felling, and their dependent changes (tdc) with the breeding success of I. typographus . Twenty trees were classified according to age (34/90 years) and crown density (high/intermediate/low). Water content was higher in young trees than in old trees, higher in the crown than at breast height, and decreased significantly within the 8-week study period. In old trees, breeding success, length of mother galleries and SO₄²⁻ were significantly higher, while total protein, NO₃⁻ and water content were significantly lower than in young trees. Trees with intermediate crown density provided the best breeding success for I. typographus and had significantly higher arginine content and C/N ratio as well as low amounts of phosphate and glutamine. During the period of bark beetle breeding, total sulphur, glutathione, protein, NO₃⁻, aspartate, glutamine, glutamate, arginine and γ-aminobutyrate decreased significantly. The results support previous investigations that I. typographus develops best in physiologically weakened trees.     

Plasmid-determined resistance to chromate in Pseudomonas aeruginosa

Abstract Resistance to chromate in five independent Pseudomonas aeruginosa clinical isolates was transferred by conjugation to P. aeruginosa strain PU21. All chromate-resistant transconjugants contained large plasmids that also conferred resistance to inorganic mercury. One of these plasmids, pUM505, increased the resistance to CrO₄²⁻ and decreased the accumulation of intracellular ⁵¹CrO₄²⁻ by the host cells as compared to the plasmidless strain PU21.     

The controlling influence of cell-surface electrical potential on the uptake and toxicity of selenate (SeO42–)

Root elongation in wheat seedlings ( Triticum aestivum L. cv. Atlas 66) was inhibited by micromolar activities of SeO₄^2–. SeO₄^2– inhibition was enhanced by supplementation of the rooting medium with CaCl₂, MgCl₂, SrCl₂, or the reduction of pH. These solute treatments, as well as the addition of tris (ethylenediamine)cobalt³⁺, enhanced the uptake of Se by the roots. The results are interpreted to reflect an elevated PM-surface activity of SeO₄^2– caused by solute-induced reductions of plasma membrane (PM) surface negativity. (PM-surface electrical potential is sometimes measured electrophoretically as the zeta potential.) This study complements an extensive literature documenting the suitability of an electrostatic model (Gouy-Chapman-Stern), based almost entirely upon experiments with cations rather than anions. The close correspondence among uptake, intoxication, and model-computed SeO₄^2– activity at the PM surface adds credibility to the model and its evaluated parameters. The model may be useful for the interpretation of other plant-anion interactions, and phosphate and sulphate nutrition in acidic soils are considered as examples.     

A LIGHT-SCATTERING TECHNIQUE FOR THE STUDY OF THE PERMEABILITY OF RAT BRAIN SYNAPTOSOMES IN VITRO

Abstract— (1) Swelling of synaptosomes was measured spectrophotometrically by recording changes in extinction at 520 nm.
(2) Synaptosomes behaved as osmometers in NaCl solutions. When the tonicity of the medium was changed, synaptosome volume changed in accordance with Boyle and van't Hoff's Law. These changes were reversed on restoring the tonicity of the medium.
(3) The rate at which a solute entered the synaptosome was determined from the rate of swelling in the presence of that solute. Permeability of synaptosomes to non-electrolytes was in the order glucose ≪ glycerol < thiourea = formamide < propylene glycol = dimethylsulphoxide.
(4) Synaptosomes were freely permeable to ammonium and acetate ions and impermeable to Ca²⁺, Mg²⁺, PO₄²⁻, SO₄²⁻ and oxalate ions.     

Histological changes in the gill, kidney and liver of Lahontan cutthroat trout, Salmo clarki henshawi, living in lakes of different salinity-alkalinity

Lahontan cutthroat trout thrive in saline-alkaline lakes, where other trout species often cannot survive. We examined Lahontan cutthroat trout from nine lakes in which salinity and alkalinity ranged from about 90 to 12 000 mg1⁻¹ and 60 to 3500mgl⁻¹ as HCO₃⁻ respectively, for sublethal histological changes in gill, kidney, and liver tissues. Gill chloride cell hyperplasia, gill lamellar epithelial separation, kidney glomerular swelling, blood congestion in kidneys, and deposition of hyalin droplets in kidney glomeruli, tubules, and hemopoietic tissues were the histological alterations statistically associated with differences in lakewater chemistry.
Deposition of hyalin in kidney tubules was the only histological change judged pathological and whose severity appeared sufficient to jeopardize normal organ function. Differences in lakewater chemistry explained nearly 90% of the variability observed in severity of tubular hyalin degeneration, and SO₄²⁻ was the ion most positively correlated with increasing tubular hyalin. Our results suggest that Lahontan cutthroat trout will develop slight to moderate hyalin degeneration in kidney tubules if stocked into lakes where salinity and SO₄²⁻ concentrations equal or exceed 5000 mgl⁻¹ and 2000mgl⁻¹, respectively.     

Root ammonium transport efficiency as a determinant in forest colonization patterns: an hypothesis

Ratios of ammonium (NH₄⁺) to nitrate (NO₃^–) in soils are known to increase during forest succession. Using evidence from several previous studies, we hypothesize that a malfunction in NH₄⁺ transport at the membrane level might limit the persistence of early successional tree species in later seral stages. In those studies, ¹³N radiotracing was used to determine unidirectional fluxes and pool sizes of NH₄⁺ and NO₃^– in seedlings of the late-successional species white spruce ( Picea glauca ) and in the early successional species Douglas-fir ( Pseudotsuga menziesii var. glauca ) and trembling aspen ( Populus tremuloides ). At high external NH₄⁺, the two early successional species accumulated excessive NH₄⁺ in the root cytosol, and exhibited high-velocity, low-efficiency (15% to 22%), membrane fluxes of NH₄⁺. In sharp contrast, white spruce had low cytosolic NH₄⁺ accumulation, and lower-velocity but much higher-efficiency (65%), NH₄⁺ fluxes. Because these divergent responses parallel known differences in tolerance and toxicity to NH₄⁺ amongst these species, we propose that they constitute a significant driving force in forest succession, complementing the discrimination against NO₃^– documented in white spruce (Kronzucker et al. 1997).     

Response of nitrogen metabolism to boron toxicity in tomato plants

Boron (B) toxicity has become important in areas close to the Mediterranean Sea where intensive agriculture has been developed. The objective of this research was to study the effects of B toxicity (0.5 m m and 2.0 m m B) on nitrogen (N) assimilation of two tomato cultivars that are often used in these areas. Leaf biomass, relative leaf growth rate (RGR_L), concentration of B, nitrate (NO₃⁻), ammonium (NH₄⁺), organic N, amino acids and soluble proteins, as well as nitrate reductase (NR), nitrite reductase (NiR), glutamine synthase (GS), glutamate synthetase (GOGAT) and glutamate dehydrogenase (GDH) activities were analysed in leaves. Boron toxicity significantly decreased leaf biomass, RGR_L, organic N, soluble proteins, and NR and NiR activities. The lowest NO₃⁻ and NH₄⁺ concentration in leaves was recorded when plants were supplied with 2.0 m m B in the root medium. Total B, amino acids, activities of GS, GOGAT and GDH increased under B toxicity. Data from the present study prove that B toxicity causes inhibition of NO₃⁻ reduction and increases NH₄⁺ assimilation in tomato plants.     

Kinetics of FeS-mediated denitrification in sediments from the Camargue (Rhone delta, southern France)

Abstract Denitrification rates were measured in sediments after the addition of different concentrations of FeS. A decrease of the denitrification rate was observed when high concentrations of ferrous iron (> 10 mM) were present. In the experiments with no significant concentrations of free Fe²⁺, the relationship between NO₃⁻ reduction and FeS concentration followed Michaelis and Menten kinetics. The maximum rate was 0.273 mmol l⁻¹ d⁻¹, 6 times as much as the basal rate 0.046 mmol l⁻¹ d⁻¹, which was attributed to organic matter; the K_s was 1.45 mM FeS. The stoichiometry of the overall reaction involving NO₃⁻ reduction and the concomitant S²⁻ oxidation was also investigated. Measured ΔS/ΔN ratios ranged between 0.55 and 0.64, with ΣH₂S + SO₄²⁻ changing less than 10%. These values agree with the theoretically expected value of 0.56.     

Mice Deficient in Group IV Cytosolic Phospholipase A2 Are Resistant to MPTP Neurotoxicity

Abstract: Phospholipase A₂ (PLA₂) enzymes are critical regulators of prostaglandin and leukotriene synthesis, and they may also play an important role in the generation of intracellular free radicals. The group IV cytosolic form of phospholipase A₂ (cPLA₂) is regulated by changes in intracellular calcium concentration, and the enzyme preferentially acts to release arachidonic acid esterified at the sn -2 position of phospholipids. We examined the susceptibility of mice carrying a targeted mutation of the cPLA₂ gene to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity. Mutant mice have no functional cPLA₂ activity. Mice that were homozygous for the mutation (cPLA₂^−/−) were significantly resistant to MPTP-induced dopamine depletion as compared with littermate control (cPLA₂^+/+) and heterozygous mice (cPLA₂^+/−). These findings provide evidence that cPLA₂ plays a role in MPTP neurotoxicity and suggest that cPLA₂ may play a role in the development of Parkinson's disease in humans.     

Light activates H2 15O flow in rice: Detailed monitoring using a positron-emitting tracer imaging system (PETIS)

Water (H₂ ¹⁵O) translocation from the roots to the top of rice plants ( Oryza saliva L. cv. Nipponbare) was visualized over time by a positron-emitting tracer imaging system (PETIS). H₂ ¹⁵O flow was activated 8 min after plants were exposed to bright light (1 500 μmol m⁻² s⁻¹). When the light was subsequently removed, the flow gradually slowed and completely stopped after 12 min. In plants exposed to low light (500 μmol m⁻² s⁻¹), H₂ ¹⁵O flow was activated more slowly, and a higher translocation rate of H₂ ¹⁵O was observed in the same low light at the end of the next dark period. NaCl (80 m M ) and methylmercury (1 m M ) directly suppressed absorption of H₂ ¹⁵O by the roots, while methionine sulfoximine (1 m M ), abscisic acid (10 μ M ) and carbonyl cyanide m -chlorophenylhydrazone (10 m M ) were transported to the leaves and enhanced stomatal closure, reducing H₂ ¹⁵O translocation.     

Growth, photosynthesis, and respiration of Chlorella sorokiniana after N-starvation. Interactions between light, CO2 and NH4+ supply

N-sufficient cells of Chlorella sorokiniana Shihira and Krauss, strain 211/8k, absorbed NH₄⁺ under light plus CO₂ conditions, when growth occurred, but not in darkness or in the absence of CO₂, when growth was inhibited. N-sufficient cells subjected to conditions of N-starvation for a 24-h period showed a marked loss of photosynthetic activity. Upon supply of NH₄⁺, N-starved cells sufflated with CO₂ air exhibited a time-dependent recovery of photosynthetic activity, both when suspended in light and in darkness. By contrast, growth only occurred in cells suspended in light. N-starved cells absorbed NH₄⁺ in darkness, but at a lower rate than in light. All of these data suggest that dark NH₄⁺ uptake is driven by N assimilation to recover from N-starvation and that the light-dependent NH₄⁺ uptake is driven by growth, being then influenced by conditions that affect recovery or growth. Unlike CO₂ conditions, in a CO₂-free atmosphere, absorption of NH₄⁺ by N-starved cells occurred at a higher rate in darkness than in light. Accordingly, resumption of photosynthetic potential after NH₄⁺ supply occurred in darkened cells, but not in illuminated cells. Respiratory activity of N-starved cells was enhanced up to 3-fold by NH₄⁺ and 2-fold by methylammonium, with different patterns, suggesting that respiratory enzymes were affected by N-metabolism, especially through short-term control mechanisms triggered by the expenditure of metabolic energy involved in N-metabolism.     

SELECTIVE PERMEABILITY OF THE EXTRACELLULAR ENVELOPE OF THE MICROALGA SPONDYLOSIUM PANDURIFORME (CHLOROPHYCEAE) AS REVEALED BY ELECTRON PARAMAGNETIC RESONANCE

The aim of this work was to investigate the role of the polysaccharide sheath of the microalga Spondylosium panduriforme (Chlorophyceae, Desmidiaceae) in the selective permeability and transport of molecules into the interior of the cell. We have used the electron paramagnetic resonance (EPR) technique applied to a variety of spin labels of a hydrophobic nature with different substitutents on the ring (−OH, =O, −N=C=S, −NH₃⁺, and others). The spin label EPR signals were destroyed as a consequence of metabolic processes once the spin probes had entered the cells. The decay time of the EPR signal was regulated by the diffusion mechanism across the polysaccharide sheath, cell wall, and membrane. To discriminate the effect of the polysaccharide sheath from that of the cell wall and membrane, the polysaccharide sheath was removed by ultrasonic treatment. The decay times for the cells without capsule were faster than those for intact cells, and a possible mechanism of interaction involving hydrogen bonds between the spin labels and the −OH groups of the polysaccharide sheath is presented. These were expressed by their diffusion and friction coefficients as derived from Ficks' Second Law and the Einstein-Stokes equation and were summarized in terms of diffusion coefficients ( D ₁) for the capsule medium in the order: =O < −OH < −phe < −H < −N=C=S; and for cell wall and membrane ( D ₂): −OH < −H < =O < −NH₃⁺≅−phe < −N=C=S. For the friction coefficients ( f ₁ and f ₂), the order was inverted. These results suggest the capsule plays a role in selectivity as a result of polar interactions with the spin labels.     

Effects of hypochlorite disinfection on the response of barley aleurone layers to gibberellic acid

Although the use of hypochlorite to disinfect seeds is widespread, the effects on tissues isolated from them have been largely ignored. Disinfection of barley ( Hordeum vulgare L. cv. Himalaya) half-seeds with hypochlorite solutions of ≥1.0% (w/v) available chlorine caused the pericarp to separate from the underlying tissues. Aleurone layers isolated from these grains had lower rates of oxygen consumption and released significantly less protein, PO₄³⁻ Mg²⁺, K⁺ and amylase (EC 3.2.1.1) into the medium in response to gibberellic acid (GA₃) than layers isolated from grains disinfected with a 0.1% hypochlorite solution. Disinfection with 1.0% hypochlorite also quantitatively altered the spectrum of proteins released into the incubation medium by the layers in response to GA₃.     

Salt-induced oxidative stress mediated by activated oxygen species in pea leaf mitochondria

The effect in vivo of salt stress on the activated oxygen metabolism of mitochondria, was studied in leaves from two NaCl-treated cultivars of Pisum sativum L. with different sensitivity to NaCl. In mitochondria from NaCl-sensitive plants, salinity brought about a significant decrease of Mn-SOD (EC 1. 15. 1. 1) Cu, Zn-SOD I (EC 1. 15. 1. 1) and fumarase (EC 4. 2. 1. 2) activities. Conversely, in salt-tolerant plants NaCl treatment produced an increase in the mitochondrial Mn-SOD activity and, to a lesser extent, in fumarase activity. In mitochondria from both salt-treated cultivars, the internal H₂O₂ concentration remained unchanged. The NADH- and succinate-dependent generation of O₂^.−radicals by submitochondrial particles and the lipid peroxidation of mitochondrial membranes, increased as a result of salt treatment, and these changes were higher in NaCl-sensitive than in NaCl-tolerant plants. Accordingly, the enhanced rates of superoxide production by mitochondria from salt-sensitive plants were concomitant with a strong decrease in the mitochondrial Mn-SOD activity, whereas NaCl-tolerant plants appear to have a protection mechanism against salt-induced increased O₂^.− production by means of the induction of the mitochondrial Mn-SOD activity. These results indicate that in the subcellular toxicity of NaCl in pea plants, at the level of mitochondria, an oxidative stress mechanism mediated by superoxide radicals is involved, and also imply a function for mitochondrial Mn-SOD in the molecular mechanisms of plant tolerance to NaCl.     

An alternative explanation for the post-disturbance NO3– flush in some forest ecosystems

The appearance of soil NO₃^– after forest disturbance is commonly ascribed to a higher availability of NH₄⁺ to autotrophic nitrifiers, or to a reduction in available-C resulting in lower microbial assimilation of NO₃^–. Alternatively, it has been proposed that increasing NH₄⁺ pools following disturbance could increase net nitrification by reducing microbial assimilation of NO₃^–. Forest floor material was collected from shelterwood harvest plots which displayed both low available-C and low NH₄⁺ pools, and where previous experiments had suggested the prevalence of heterotrophic nitrification. Subsamples were amended with incremental rates of glucose-C or NH₄⁺, and gross NO₃^– transformation rates were measured by isotope dilution. Glucose-C additions had little effect on the net difference between gross NO₃^– production and consumption rates. On the other hand, NH₄⁺ additions caused gross NO₃^– consumption processes to decrease sharply, while gross NO₃^– production processes remained constant. The results suggest that NH₄⁺ can have an immediate positive effect on net nitrification rates by suppressing NO₃^– assimilation and uptake systems.     

Effects of soil frost on nitrogen net mineralization, soil solution chemistry and seepage losses in a temperate forest soil

Freezing and thawing may alter element turnover and solute fluxes in soils by changing physical and biological soil properties. We simulated soil frost in replicated snow removal plots in a mountainous Norway spruce stand in the Fichtelgebirge area, Germany, and investigated N net mineralization, solute concentrations and fluxes of dissolved organic carbon (DOC) and of mineral ions (NH₄⁺, NO₃⁻, Na⁺, K⁺, Ca²⁺, Mg²⁺). At the snow removal plots the minimum soil temperature was −5 °C at 5 cm depth, while the control plots were covered by snow and experienced no soil frost. The soil frost lasted for about 3 months and penetrated the soil to about 15 cm depth. In the 3 months after thawing, the in situ N net mineralization in the forest floor and upper mineral soil was not affected by soil frost. In late summer, NO₃⁻ concentrations increased in forest floor percolates and soil solutions at 20 cm soil depth in the snow removal plots relative to the control. The increase lasted for about 2–4 months at a time of low seepage water fluxes. Soil frost did not affect DOC concentrations and radiocarbon signatures of DOC. No specific frost effect was observed for K⁺, Ca²⁺ and Mg²⁺ in soil solutions, however, the Na⁺ concentrations in the upper mineral soil increased. In the 12 months following snowmelt, the solute fluxes of N, DOC, and mineral ions were not influenced by the previous soil frost at any depth. Our experiment did not support the hypothesis that moderate soil frost triggers solute losses of N, DOC, and mineral ions from temperate forest soils.