Evaluation of sulphur cycling in managed forest stands by means of stable S-isotope analysis

Sulphur cycling was evaluated in a 20 to 60 year old Norway spruce (Picea abies L. Karst) ecosystem in the Black Forest near Schluchsee, SW Germany, by means of stable sulphur isotope analysis.Soil and plant material were analysed for S-content and S-isotopic composition to gather information on the S-distribution in the ecosystem. Two out of three adjacent watershed areas, highly comparable to each other were fertilized with MgSO₄ and (NH₄)₂SO₄ respectively, where sulphate was enriched in the ³⁴S-isotope compared to the sulphur present in the ecosystem. As the fertilizer S served as a tracer, comparison of the S-isotopic composition of total and inorganic S in the soil and S in spruce needles from both the treated and the control sites led to new information of S-turnover processes.The S-isotopic composition of spruce needles changed markedly after the fertilizer application. Within half a year a shift towards the S-isotopic composition of the fertilizers sulphate indicated uptake of the sulphate by the trees, although this uptake did not become visible with the S content of the needles.Regarding the soil, a shift in the S-isotopic composition of the total sulphur was not that striking as with the needles, although the phosphate extractable sulphate showed a clear shift towards the S-isotopic composition of the fertilizer sulphate.     

Effect of nitrogen nutrition on the export of sulphur from leaves in soybean

Soybean plants were grown in complete solution for 33 days and then transferred to medium containing inadequate sulphur (5 t M) and nitrogen at 15, 7.5, 2 or 0.25 mt M. In mature leaves (L1 and L2), and leaves that were 70% expanded at day 33 (L3), the net loss of sulphur over the ensuing 25 days was inversely related to the level of nitrogen nutrition. Leaf 5, which formed during the study period, exhibited complementary characteristics; the increase in the sulphur content was inversely related to the level of nitrogen nutrition even though low nitrogen nutrition supported less growth. L4, which was 31% expanded at day 33, exhibited intermediate characteristics. 35S-Labelled sulphate was supplied to all of the plants for 48 h at day 31 and was distributed principally to L3 at day 33. During early development, L5 became heavily labelled but, at low nitrogen nutrition, the massive import of total sulphur into L5 during the late stages of development was not accompanied by a commensurate increase in 35S-label, indicating that redistribution of soluble sulphur from mature leaves was not involved. The loss of sulphur from mature leaves was parallelled by similar changes in nitrogen at all levels of nitrogen nutrition. Collectively, the data suggest that a common mechanism, presumably proteolysis, is involved in the export of sulphur and nitrogen from mature leaves and that this process is inhibited at high levels of nitrogen nutrition, even under conditions of sulphur deficiency.     

Changes in the chemical content of birch (<Emphasis Type="Italic">Betula pendula </Emphasis>Roth) leaves in the air polluted Krusne hory mountains

Analyses of birch leaves (Betula pendula Roth) sampled from trees in the air polluted Krusne hory mountains and Decin sandstone highlands were performed in 1995 and 1998. The results revealed a connection between reduction of air pollutants and overall decrease in sulphur and nitrogen content of the leaves. Simultaneously, in many stands an increase in calcium content and a decrease in contents of phosphorus, magnesium and potassium occurred. The content of microelements and non-nutrient elements did not change.     

Combustion losses of sulphur from conifer foliage: Implications of chemical form and soil nitrogen status

The proportion of total sulphur lost during combustion (600 °C) of Douglas-fir (Pseudotsuga menziesii) foliage is reduced from> 90% to 65–70% as the SO₄-S concentration increases from 10% to 45–50% of the total S content. Foliar SO₄-S content is decreased by improvement of plant nitrogen status, suggesting that alterations to soil N availability may influence S transfer to the atmosphere during biomass burning.     

Extractable sulphate and organic sulphur in soils and their availability to plants

Ten soils collected from the major arable areas in Britain were used to assess the availability of soil sulphur (S) to spring wheat in a pot experiment. Soils were extracted with various reagents and the extractable inorganic SO₄-S and total soluble S(SO₄-S plus a fraction of organic S) were determined using ion chromatography (IC) or inductively-coupled plasma atomic emission spectrometry (ICP-AES), respectively. Water, 0.016 M KH₂PO₄, 0.01 M CaCl₂ and 0.01 M Ca(H₂PO₄)₂ extracted similar amounts of SO₄-S, as measured by IC, which were consistently smaller than the total extractable S as measured by ICP-AES. The amounts of organic S extracted varied widely between different extractants, with 0.5 M NaHCO₃ (pH 8.5) giving the largest amounts and 0.01 M CaCl₂ the least. Organic S accounted for approximately 30–60% of total S extracted with 0.016 M KH₂PO₄ and the organic C:S ratios in this extract varied typically between 50 and 70. The concentrations of this S fraction decreased in all soils without added S after two months growth of spring wheat, indicating a release of organic S through mineralisation. All methods tested except 0.5 M NaHCO₃-ICP-AES produced satisfactory results in the regression with plant dry matter response and S uptake in the pot experiment. In general, 0.016 M KH₂PO₄ appeared to be the best extractant and this extraction followed by ICP-AES determination was considered to be a good method to standardise on.     

Stable sulphur isotopes in plants: a review

The determination of the natural abundances of stable isotopes has become a useful method by which to study the transformations of elements in biological and ecological studies as well as to investigate the mechanisms of chemical reactions. Unlike carbon and nitrogen isotopes, however, stable sulphur isotopes are used infrequently, and their potential as tracers in biochemical and physiological studies are only beginning to be realized. This review provides an overview of research involving stable sulphur isotopes in studies of plant metabolism and pollution. Topics discussed include the mechanisms and accompanying isotopic fractionations involved during the uptake and assimilation of inorganic sulphur compounds by plants, the utility of plants as bioindicators of environmental sulphur pollution, and the emission of isotopically light H₂S by plants in response to high concentations of sulphur. Future advances in the field are proposed.     

Interactive effects of elevated atmospheric CO2, mycorrhization and drought on long-distance transport of reduced sulphur in young pedunculate oak trees (Quercus robur L.)

Pedunculate oak (Quercus robur L.) was germinated and grown under nutrient non-limiting conditions for a total of 10–15 weeks at ambient CO₂ concentration and 1100 μmol mol^–1 CO₂ either in the presence or the absence of the mycorrhizal fungus Laccaria laccata. Half of the oak trees of these treatments were exposed to drought during final growth by suspending the water supply for 21 d. Mycorrhization and elevated atmospheric CO₂ each enhanced total plant biomass per tree. Whereas additional biomass accumulation of trees grown under elevated CO₂ was mainly attributed to increased growth of lateral roots, mycorrhization promoted shoot growth. Water deficiency reduced biomass accumulation without affecting relative water content, but this effect was more pronounced in mycorrhizal as compared to non-mycorrhizal trees. Elevated CO₂ partially prevented the development of drought stress, as indicated by leaf water potential, but did not counteract the negative effects of water deficiency on growth during the time studied. Enhanced biomass accumulation requires adaption in protein synthesis and, as a consequence, enhanced allocation of reduced sulphur produced in the leaves to growing tissues. Therefore, allocation of reduced sulphur from oak leaves was studied by flap-feeding radiolabelled GSH, the main long-distance transport form of reduced sulphur, to mature oak leaves. Export of radiolabel proceeded almost exclusively in basipetal direction to the roots. The rate of export of radioactivity out of the fed leaves was significantly enhanced under elevated CO₂, irrespective of mycorrhization. A higher proportion of the exported GSH was transported to the roots than to basipetal stem sections under elevated CO₂ as compared to ambient CO₂. Mycorrhization did not affect ³⁵S export out of the fed leaves, but the distribution of radiolabel between stem and roots was altered in preference of the stem. Trees exposed to drought did not show appreciable export of the ³⁵S radioactivity fed to the leaves when grown under ambient CO₂. Apparently, drought inhibited basipetal transport of reduced sulphur at the level of phloem loading and/or phloem transport. Elevated CO₂ seemed to counteract this effect of drought stress to some extent, since higher leaf water potentials and improved ³⁵S export out of the fed leaves was observed in oak trees exposed to drought and elevated CO₂ as compared to trees exposed to drought and ambient CO₂.     

Effects of sulphur dioxide fumigation on soil system and growth behaviour ofVicia faba plants

Summary To determine the possible changes in soil characteristics and plant growth resulting from SO₂-exposure, field grownVicia faba plants were exposed to 0.5 ppm SO₂ for 4 h daily from 40 to 100 days of their ages. The major effects of SO₂ on soil system includes increases in contents of total sulphur, organic carbon and exchangeable Al; lowering of pH and decreases in contents of N, P and exchangeable K and Ca. Besides, SO₂ interfered with nutrient uptake and plant growth leading to reductions in lengths of root and shoot, numbers of leaves, nodules and pods, phytomass accumulation and net primary productivity.     

Decline in gypsy moth (Lymantria dispar) performance in an elevated CO2 atmosphere depends upon host plant species

Plant species differ broadly in their responses to an elevated CO₂ atmosphere, particularly in the extent of nitrogen dilution of leaf tissue. Insect herbivores are often limited by the availability of nutrients, such as nitrogen, in their host plant tissue and may therefore respond differentially on different plant species grown in CO₂-enriched environments. We reared gyspy moth larvae (Lymantria dispar) in situ on seedlings of yellow birch (Betula allegheniensis) and gray birch (B. populifolia) grown in an ambient (350 ppm) or elevated (700 ppm) CO₂ atmosphere to test whether larval responses in the elevated CO₂ atmosphere were species-dependent. We report that female gypsy moths (Lymantria dispar) reared on gray birch (Betula populifolia) achieved similar pupal masses on plants grown at an ambient or an elevated CO₂ concentration. However, on yellow birch (B. allegheniensis), female pupal mass was 38% smaller on plants in the elevated-CO₂ atmosphere. Larval mortality was significantly higher on yellow birch than gray birch, but did not differ between the CO₂ treatments. Relative growth rate declined more in the elevated CO₂ atmosphere for larvae on yellow birch than for those on gray birch. In preference tests, larvae preferred ambient over elevated CO₂-grown leaves of yellow birch, but showed no preference between gray birch leaves from the two CO₂ atmospheres. This differential response of gypsy moths to their host species corresponded to a greater decline in leaf nutritional quality in the elevated CO₂ atmosphere in yellow birch than in gray birch. Leaf nitrogen content of yellow birch dropped from 2.68% to 1.99% while that of gray birch leaves only declined from 3.23% to 2.63%. Meanwhile, leaf condensed tannin concentration increased from 8.92% to 11.45% in yellow birch leaves while gray birch leaves only increased from 10.72% to 12.34%. Thus the declines in larval performance in a future atmosphere may be substantial and host-species-specific.     

Reduced sulphur sources favour HgII reduction during anoxygenic photosynthesis by Heliobacteria

The consumption of rice has become a global food safety issue because rice paddies support the production of high levels of the potent neurotoxin, methylmercury. The production of methylmercury is carried out by chemotrophic anaerobes that rely on a diversity of terminal electron acceptors, namely sulphate. Sulphur can be a limiting nutrient in rice paddies, and sulphate amendments are often used to stimulate crop production, which can increase methylmercury production. Mercury (Hg) redox cycling can affect Hg methylation by controlling the delivery of inorganic Hg substrates to methylators in anoxic habitats. Whereas sulphur is recognized as a key substrate controlling methylmercury production, the controls sulphur exerts on other microbe‐mediated Hg transformations remain poorly understood. To explore the potential coupling between sulphur assimilation and anaerobic Hg^II reduction to Hg⁰, we studied Heliobacillus mobilis, a mesophilic anoxygenic phototroph representative from the Heliobacteriacea family originally isolated from a rice paddy. Here, we tested whether the redox state of the sulphur sources available to H. mobilis would affect its ability to reduce Hg^II. By comparing Hg⁰ production over a redox gradient of sulphur sources, we demonstrate that phototrophic Hg^II reduction is favoured in the presence of reduced sulphur sources such as thiosulphate and cysteine. We also show that cysteine exerts dynamic control on Hg cycling by affecting not only Hg's bioavailability but also its abiotic photoreduction under low light conditions. Specifically, in the absence of cells we show that organic matter (as yeast extract) and cysteine are both required for photoreduction to occur. This study offers insights into how one of the most primitive forms of photosynthesis affects Hg redox transformations and frames Heliobacteria as key players in Hg cycling within paddy soils, forming a basis for management strategies to mitigate Hg accumulation in rice.     

Isolation of the sulphur reductase and reconstitution of the sulphur respiration of Wolinella succinogenes

Wolinella succinogenes can grow at the expense of sulphur reduction by formate. The enzymes involved in the catalysis of this catabolic reaction have been investigated. From the results the following conclusions are drawn: 1. The enzyme isolated as a sulphide dehydrogenase from the cytoplasmic membrane of W. succinogenes is the functional sulphur reductase that operates in the electron transport from formate to sulphur. 2. The enzyme (M_r 200,000) consists essentially of one type of subunit with the M_r 85,000 and contains equal amounts of free iron and sulphide (120 mol/g protein), but no heme. It represents the first functional sulphur reductase ever isolated. 3. The electron transport chain catalyzing sulphur reduction by formate consists merely of formate dehydrogenase and sulphur reductase. A lipophilic quinone which mediates the transfer of electrons between enzymes in other chains, is apparently not involved. This is the first known example of a phosphorylative electron transport chain that operates without a quinone. 4. The same formate dehydrogenase appears to operate in the electron transport both with sulphur and with fumarate as the terminal electron acceptor in W. succinogenes.Abbreviations DMN 2,3-Dimethyl-1,4-naphthoquinone - DTT dithiothreitol - MK menaquinone (vitamin K₂) - PMSF phenylmethane sulfonylfluoride - Tricine N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]-glycine - Tea triethanolamine - Hepes 4-(2-hydroxyethyl)-1-piperazineethane sulfonate Dedicated to Professor F. Schneider (Philipps-Universität Marburg) on the occasion of his 60th birthday     

Effects of sulphur dioxide preservation of flax straw on fibre components and microbial colonisation during dew-retting

The changes in flax straw structure when treated with sulphur dioxide (SO₂) and consequent effect on dew-retting of the treated straw were investigated. The active oxoanions released by sulfurous acid degraded phenolics bound to the polysaccharides and lipids leading to an increase in water-soluble carbohydrates and polyphenols. As a result, proportions of hemicellulose, nitrogen, sulphur, ash and caustic weight loss were higher in treated straw while lipid was reduced. The coarseness of treated fibre was possibly caused by in situ dryback of the hydrolysed polysaccharides. The deposition of salts containing high concentrations of sulphur, calcium and potassium in the vicinity of the fibre bundles and epidermal layer was observed and investigated. In a dew-retting trial comprising SO₂-treated and untreated samples, the SO₂ treatment delayed microbial colonisation and, consequently, retting by nearly 2 wk compared to the controls. The changes in pH, fungal population and polysaccharide degrading enzymes were monitored. The differences in fibre yield, fineness and strength are presented.     

Mobilization of sulphur in soybean cotyledons during germination

Soybean seeds ( Glycine max L. cv , Stephens) contain a large amount of sulphur (ca 40 μ mol seed⁻¹), mostly in the insoluble fraction in the cotyledons. During germination in nutrient solution lacking sulphur the amount of insoluble sulphur decreases to very low levels. This is accompanied by a transitory increase in the pool of soluble sulphur which then declines. All of the sulphur lost from the cotyledons is quantitatively recovered in the seedling. In the short term, the root and the stem are the most important sinks for sulphur from the cotyledons but as growth proceeds the shoot becomes the dominant sink for remobilized sulphur. Within the shoot most of the sulphur is recovered in leaves L1 and L2. The growth of L3 and, to a lesser extent, L2, was retarded due to sulphur insufficiency. The cotyledons of plants treated with 20 μ M sulphate also exhibited mobilization of sulphur from the insoluble fraction except that the maximum rate of loss of sulphur occurred somewhat later. Plants grown with sulphate exhibited a net gain of sulphur and did not exhibit sulphur insufficiency. In these plants, endogenous sulphur from the cotyledons was directed into L1–L3 and this sulphur remained within these leaves for the duration of the experiment. The delivery of exogenous sulphur (supplied as [³⁵S]sulphate via the roots) to the leaves increased with leaf number. In leaves L1–L3, the level of exogenous sulphur in any one leaf declined with time, indicating that this sulphur was remobilized and did not mix with the sulphur derived from the cotyledons. It was concluded that the cotyledons are an important source of sulphur to support early plant growth and development of soybean.     

Effect of high rates of sulphur on the CaCO3 and CaSO4 content of calcareous soils

Samples of two calcareous soils from central Iraq were amended with 2, 6 and 10 mg g⁻¹ soil of agriculturegrade sulphur and incubated for 4, 8 and 12 weeks at 30°C and 70% water-holding capacity. At the end of each incubation period, soils were analyzed for sulphate, CaCO₃ and CaSO₄. The sulphate content of the soils increased, whereas the CaCO₃ content decreased, with increasing S levels and incubation time. The gypsum formed was in proportion to the sulphur oxidised and the calcium released from CaCO₃.     

Distribution of sulphur forms in soils from beech and spruce forests of Mont Lozère (France)

From a quantitative inventory of sulphur forms and sulphur budget, the relation between the distribution of the various sulphur forms and the sulphate fluxes in three soil profiles has been addressed. These profiles are located in two forested watersheds at Mont Lozère. One has been sampled in a beech forest and the other two in a spruce forest and in a harvested plot of this spruce forest, respectively. The mean annual input-output budgets showed a sulphur immobilization in the soil cover of the three plots. In the preserved spruce forest plot, because of larger dry depositions, the sulphur immobilization is much greater than in other plots and occurs essentially in the B horizons. In the other two profiles, the dominant immobilization occurs in the parent material.The total sulphur content is very high in the forest floor reaching 2065 g S g^-1 in the litter of one of the soils under spruce. In the organo-mineral horizons of soils under spruce, the total sulphur content decreases with depth and ranges from 310 to 520 g S g^-1 in the A horizons to 100–200 g S g^-1 in the parent material. In the profile under beech, the total sulphur content is lower except in the parent material. In all cases, the organic sulphur is the major part of sulphur often representing more than 90% of total sulphur. In organo-mineral soil horizons of the spruce forest, the part of the sulphateesters is more important than in the soil of the beech forest, probably related to the different nature of the microbial activity in the spruce forest. In contrast, the humification processes are more efficient in the soil under beech, which can be due to the greater input of organic sulphur by litterfall. It appears that the dominant organic sulphur form varies as a function of microbial ecology and sulphate flux. The maximum of the inorganic sulphate is located at the base of the B horizons in the soil of the spruce forest and in the parent material of the soil under beech. In these horizons, the high content of inorganic sulphate can be related to the higher amounts of amorphous Fe and Al phases.     

Removal of H2S by the purple sulphur bacterium Ectothiorhodospira shaposhnikovii

When Ectothiorhodospira shaposhnikovii VKM B-1525 was used for desuphurization of biogas in the laboratory and in a pilot plant, there was complete oxidation of H₂S, the main product being elemental sulphur. The advatage of this culture over green bacteria is discussed.M.B. Vainshtein, G.I. Gogotova and N.-J. Heinritz are with the Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Moscow Region, 142292 Russia     

Copper toxicity in Chinese cabbage is not influenced by plant sulphur status,but affects sulphur metabolism‐related gene expression and the suggested regulatory metabolites

The toxicity of high copper (Cu) concentrations in the root environment of Chinese cabbage (Brassica pekinensis) was little influenced by the sulphur nutritional status of the plant. However, Cu toxicity removed the correlation between sulphur metabolism‐related gene expression and the suggested regulatory metabolites. At high tissue Cu levels, there was no relation between sulphur metabolite levels viz. total sulphur, sulphate and water‐soluble non‐protein thiols, and the expression and activity of sulphate transporters and expression of APS reductase under sulphate‐sufficient or‐deprived conditions, in the presence or absence of H₂S. This indicated that the regulatory signal transduction pathway of sulphate transporters was overruled or by‐passed upon exposure to elevated Cu concentrations.     

The mechanism of photosystem‐II inactivation during sulphur deprivation‐induced H2 production in Chlamydomonas reinhardtii

Sulphur limitation may restrain cell growth and viability. In the green alga Chlamydomonas reinhardtii, sulphur limitation may induce H₂ production lasting for several days, which can be exploited as a renewable energy source. Sulphur limitation causes a large number of physiological changes, including the inactivation of photosystem II (PSII), leading to the establishment of hypoxia, essential for the increase in hydrogenase expression and activity. The inactivation of PSII has long been assumed to be caused by the sulphur‐limited turnover of its reaction center protein PsbA. Here we reinvestigated this issue in detail and show that: (i) upon transferring Chlamydomonas cells to sulphur‐free media, the cellular sulphur content decreases only by about 25%; (ii) as demonstrated by lincomycin treatments, PsbA has a significant turnover, and other photosynthetic subunits, namely RbcL and CP43, are degraded more rapidly than PsbA. On the other hand, sulphur limitation imposes oxidative stress early on, most probably involving the formation of singlet oxygen in PSII, which leads to an increase in the expression of GDP‐L‐galactose phosphorylase, playing an essential role in ascorbate biosynthesis. When accumulated to the millimolar concentration range, ascorbate may inactivate the oxygen‐evolving complex and provide electrons to PSII, albeit at a low rate. In the absence of a functional donor side and sufficient electron transport, PSII reaction centers are inactivated and degraded. We therefore demonstrate that the inactivation of PSII is a complex and multistep process, which may serve to mitigate the damaging effects of sulphur limitation.