Sulphur oxidation by soil fungi including some species of mycorrhizae and wood-rotting basidiomycetes

Abstract The mycorrhizal fungi Amanita muscaria, Paxillus involutus, Hymenoscyphus ericae, Pisolithus tinctorius, Rhizopogon roseolus , and Suillus bovinus oxidized elemental sulphur to thiosulphate and sulphate in vitro. In some, but not all cases, tetrathionate was also formed. Limited oxidation of elemental sulphur by R. roseolus also occurred when growing in association with Pinus contorta in unsterilized peat. Although yeasts capable of oxidizing sulphur could not be isolated from a wide range of soils, a yeast-like fungus ( Monilia sp.) isolated from deciduous woodland soil oxidized elemental sulphur to sulphate, forming thiosulphate, but not tetrathionate. This fungus also oxidized tetrathionate to sulphate but showed only limited ability to oxidize thiosulphate to tetrathionate. Both Aspergillus niger and Trichoderma harzianum oxidized elemental sulphur in mixed culture with Mucor flavus . Larger amounts of sulphate were initially formed in mixed, compared to single culture; but by week 5 of the incubation period sulphate formation was greatest in single culture. The wood-rotting fungi, Hypholoma fasciculare and Phanerochaete velutina showed a limited ability to oxidize elemental sulphur in vitro but were incapable of oxidizing the element when growing as mycelial cords in non-sterilized soils. The relevance of these results to the possibility that fungi play a role in sulphur oxidation in soils is commented upon.     

The effect of sodium chloride and sulphate on sulphur oxidation in soil

Summary The effect of sodium chloride on sulphur oxidation in Terra Rossa and Rendzina soils was studied by incubation and perfusion techniques. Sulphur oxidation was observed at concentrations up to 8 per cent NaCl, but was completely arrested at 10 per cent sodium chloride. Sodium chloride caused a delay in the onset of sulphur oxidation, its rate being only slightly affected. A relationship between sulphate appearance and decrease in pH was observed only in sulphur-amended Terra Rossa soil. Under optimal conditions, 53 and 54 per cent of added sulphur (5000 ppm) was recovered as SO₄-S from the Terra Rossa and Rendzina soils, respectively. This maximal level of sulphate production was only slightly affected by the addition of sulphate up to 3000 ppm S.It was concluded that inhibition in further sulphur oxidation was not caused by sulphate accumulation.     

Dynamics of oxidation of inorganic sulphur compounds in upper soil horizons of spruce forests

Dynamics of oxidation of inorganic sulphur compounds to sulphate by the soil of spruce forests was investigated. Sulphide, sulphite and thiosulphate are oxidized to sulphate at a maximal rate at the beginning of the reaction, oxidation of elemental sulphur exhibits a lag phase. Linear relationships between the amounts of the produced sulphate and concentrations of substrates in the soil could be detected. On the basis of this finding a method for comparison of the oxidative activity of various soils was proposed.     

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.     

Sulphur oxidation by a Streptomyces sp. growing in a carbon-deficient medium and autoclaved soil

Streptomyces colonies, apparently all of the same species, were isolated from a range of soils using a polysulphide medium lacking an organic carbon source. Growth on this medium, and clearing of the otherwise white, opaque overlay, suggested that the organisms were capable of growing autotrophically. However, investigation of one of these isolates showed that it was unable to fix ¹⁴CO₂ and did not possess the enzyme ribulose bisphosphate carboxylase, showing that it was incapable of autotrophic growth. The isolate oxidized elemental sulphur, thiosulphate and tetrathionate to sulphate in vitro in carbon-deficient medium, and also oxidized elemental sulphur to sulphate when inoculated into autoclaved soil supplemented with sulphur. It also oxidized polysulphide when growing on Czapek Dox and plate count agars. The isolate can therefore grow heterotrophically in both carbon-rich media and in media lacking organic carbon — presumably by scavenging organic carbon from the laboratory atmosphere. The possible role of these organisms in sulphur oxidation in soils is commented upon.     

Some factors influencing production of sulphate by oxidation of elemental sulphur and thiosulphate in upper horizons of spruce forest soils

Some factors influencing the oxidative activity of upper horizons of spruce forest soils (a mixture of fermentative and humus layers) toward intermediates of the oxidative part of the sulphur cycle were investigated. Preincubation of the soil with added cysteine, sulphide, elemental sulphur or thiosulphate was found to stimulate enzyme systems oxidating any of these compounds. Sulphite and sulphate were ineffective in this respect. The oxidation of elemental sulphur was stimulated by CaCO3, technical urea and high doses of superphosphate and potassium sulphate. It was inhibited by KH2PO4, pure urea, 40 % potassium salt, ammonium nitrate with calcium carbonate and the fertilizer NPK I. It proceeded at the highest rate at approximately 60 % capillary capacity (61 % of mass water content). Oxidation of thiosulphate was stimulated by KH2PO4, pure urea, superphosphate, potassium sulphate and only slightly by the fertilizer NPK I. It was inhibited by CaCO3, 40 % potassium salt and only slightly by ammonium nitrate with calcium carbonate. Potassium chloride, glucose and technical urea were without effect. The oxidation proceeded at the highest rate at 35 % maximal capillary capacity (48 % mass water content).     

Effect of some sulphur compounds on soil microflora of spruce rhizosphere

The effect of a long-term application of sulphite, thiosulphate and sodium sulphate on the soil microflora and spruce seedlings was investigated in a pot experiment. Sulphur compounds decreased the concentration of bacteria, including thiobacilli, increased the concentration of microscopic fungi and sulphate-reducing bacteria; they inhibited respiration, nitrification and oxidation of thiosulphate, stimulated ammonification and oxidation of elemental sulphur. In certain cases the spruce rhizosphere exhibited just the opposite effect. In the rhizosphere the sulphate-reducing bacteria were suppressed together with thiobacilli, whose unit oxidative activity increased substantially. Growth of seedlings was inhibited by sulphite and stimulated by thiosulphate and sulphate. Sulphite, the effects of which were similar to those of sulphur dioxide immissions, was the most effective compound. In regions influenced by immissions the soil is apparently intoxicated by the absorbed sulphite.     

Microbial oxidation of elemental sulphur in brown soil

Plant and Soil - Brown soil formed from loamy clay was examined for its ability to produce sulphate from added elemental suphur. At higher rates of sulphur applications the pH of a slightly acid...     

The role of soil microbes in plant sulphur nutrition

Chemical and spectroscopic studies have shown that in agricultural soils most of the soil sulphur (>95%) is present as sulphate esters or as carbon-bonded sulphur (sulphonates or amino acid sulphur), rather than inorganic sulphate. Plant sulphur nutrition depends primarily on the uptake of inorganic sulphate. However, recent research has demonstrated that the sulphate ester and sulphonate-pools of soil sulphur are also plant-bioavailable, probably due to interconversion of carbon-bonded sulphur and sulphate ester-sulphur to inorganic sulphate by soil microbes. In addition to this mineralization of bound forms of sulphur, soil microbes are also responsible for the rapid immobilization of sulphate, first to sulphate esters and subsequently to carbon-bound sulphur. The rate of sulphur cycling depends on the microbial community present, and on its metabolic activity, though it is not yet known if specific microbial species or genera control this process. The genes involved in the mobilization of sulphonate- and sulphate ester-sulphur by one common rhizosphere bacterium, Pseudomonas putida, have been investigated. Mutants of this species that are unable to transform sulphate esters show reduced survival in the soil, indicating that sulphate esters are important for bacterial S-nutrition in this environment. P. putida S-313 mutants that cannot metabolize sulphonate-sulphur do not promote the growth of tomato plants as the wild-type strain does, suggesting that the ability to mobilize bound sulphur for plant nutrition is an important role of this species.     

江淮丘陵地区油-稻轮作条件下土壤硫库变化研究

江淮丘陵地区下蜀系黄土母质发育的水稻土油2稻轮作试验表明, 油菜种植期间, 耕层土壤硫主要来源于耕层以下土层的补给, 其次是大气干湿沉降;该时期耕层土壤硫输出主要是油菜吸收, 其次是淋失.油菜种植期间耕层土壤硫输入量小于输出, 导致耕层土壤硫库下降8.76kg·hm^-2, 22%来自无机硫库的下降.水稻种植期间, 耕层土壤硫输入主要来自灌溉水, 其次是底土层的补给和大气干湿沉降;而硫输出主要是淋失, 其次是水稻吸收.耕层土壤硫输入量大于输出, 导致耕层土壤硫库增加18.69kg·hm^-2, 18%来自无机硫库的增加.全年油2稻轮作期间耕层土壤硫输入量大于输出, 导致耕层土壤硫库增加9.93kg·hm^-2, 13%来自无机硫库的增加.     

Some factors affecting the mineralization of organic sulphur in soils

Summary Factors affecting the release of sulphate from a number of eastern Australian soils were studied.All of the soils released sulphate when dried. The amounts released were influenced by the manner in which the soil was dried. Air-drying in the laboratory at 20°C released least sulphate.Sulphate was mineralized in all soils by incubation at 30°C but the amounts mineralized could not be related to soil type or any single soil property. The ratio of nitrogen mineralized: sulphur mineralized varied widely between soils and was generally appreciably greater than the ratio of total nitrogen: organic sulphur in the soils.A rapid flush of mineralization of both sulphur and nitrogen took place when some of the soils were rewetted and incubated after they had been dried in the laboratory and stored for 4 to 5 months. Following this, the rate of mineralization was similar to that in the original undried soil. During this flush, the enhancement of sulphur mineralization was relatively greater than that of nitrogen so that the ratio of nitrogen mineralized: sulphur mineralized was considerably smaller than that during later phases of the incubation or that of the original moist soil. Soils collected after they had remained dry in the field for a similar period of time did not show this type of mineralization although they had initially done so when collected moist and air-dried in the laboratory.The effects of temperature, soil moisture, toluene and formaldehyde, and the addition of calcium carbonate to soils on the mineralization of sulphur were similar to their effects on the mineralization of nitrogen.     

The removal of pyritic sulphur from coal by Leptospirillum-like bacteria

Summary The microbial oxidation of pyritic sulphur was studied in a 4.5-l airlift fermentor at pH 1.5 and 100 g/l pulp density. By microbial leaching with Leptospirillum-like bacteria 85% of the pyritic sulphur was removed within 40 days; 30% of the removed pyrite was oxidized to elemental sulphur, the rest being transformed to soluble sulphate. Accumulation of elemental sulphur could be avoided by using a mixed culture of Leptospirillum-like bacteria and Thiobacillus ferrooxidans. Apart from oxidation of elemental sulphur neither the pure nor the mixed culture showed a significant difference as to removal of pyrite.     

Effect of different moisture sequences and a short period of drying between them on the changes in sulphate levels in soils during incubation

Summary Effect of two different moisture sequences with a short period of drying between them on the changes in its sulphate levels during incubation revealed considerable immobilisation of sulphate in soils maintained either at 50% of water holding capacity or under waterlogged condition prior to drying. The extent of immobilisation was more pronounced when starch or inorganic sulphate sulphur were added. Drying resulted in release of available sulphate sulphur in the soil previously incubated at 50% of water holding capacity and the effect was more striking when starch was added. Whereas, drying caused a decrease in the amount in the soil kept under waterlogged condition. A short period of drying did release sulphate sulphur in available form.     

Effect of industrial immissions with high sulphur dioxide content on thiobacilli and oxidative activity of spruce forest soils towards inorganic sulphur compounds

Effect of industrial immissions with high sulphur dioxide content on the upper horizons of spruce forest soils in NW Bohemia was investigated. The content of sulphates, oxidative activity towards sulphide, elemental sulphur, thiosulphate and sulphite, concentration and species representation of thiobacilli in horizons F, H and A in regions highly affected by immissions (two localities) and in regions relatively less influenced (three localities) were followed. In the affected areas the sulphur content in the soil was higher, the species representation of thiobacilli was similar and their concentration was higher, the ability of the soil to oxidize thiosulphate was inhibited and oxidation of elemental sulphur was stimulated. The oxidation of sulphide and sulphite was not significantly affected by the immissions. Changes caused by immissions could be observed only in horizons F and H and did not involve horizons A.     

Oxidation of reduced sulphur compounds by intact cells of Thiobacillus acidophilus

Oxidation of reduced sulphur compounds by Thiobacillus acidophilus was studied with cell suspensions from heterotrophic and mixotrophic chemostat cultures. Maximum substrate-dependent oxygen uptake rates and affinities observed with cell suspensions from mixotrophic cultures were higher than with heterotrophically grown cells. ph Optima for oxidation of sulphur compounds fell within the pH range for growth (pH 2–5), except for sulphite oxidation (optimum at pH 5.5). During oxidation of sulphide by cell suspensions, intermediary sulphur was formed. Tetrathionate was formed as an intermediate during aerobic incubation with thiosulphate and trithionate. Whether or not sulphite is an inter-mediate during sulphur compound oxidation by T. acidophilus remains unclear. Experiments with anaerobic cell suspensions of T. acidophilus revealed that trithionate metabolism was initiated by a hydrolytic cleavage yielding thiosulphate and sulphate. A hydrolytic cleavage was also implicated in the metabolism of tetrathionate. After anaerobic incubation of T. acidophilus with tetrathionate, the substrate was completely converted to equimolar amounts of thiosulphate, sulphur and sulphate. Sulphide- and sulphite oxidation were partly inhibited by the protonophore uncouplers 2,4-dinitrophenol (DNP) and carbonyl cyanide m-chlorophenylhydrazone (CCCP) and by the sulfhydryl-binding agent N-ethylmaleimide (NEM). Oxidation of elemental sulphur was completely inhibited by these compounds. Oxidation of thiosulphate, tetrathionate and trithionate was only slightly affected. The possible localization of the different enzyme systems involved in sulphur compound oxidation by T. acidophilus is discussed.     

Oxidation of micronized elemental sulphur in soil

The rate of oxidation of micronized elemental sulphur in three soils was measured over a range of temperatures between 2 and 20°C. Temperature had a marked effect with a Q₁₀ (temperature coefficient) between 1.9–3.1. The period for 50% oxidation varied between 6–10 days at 20°C to between 36–42 days at 2°C. All the oxidation curves showed an initial lag. At 20°C the oxidation rate was four times that of flowers of sulphur and was related to the smaller particle size. Additives (wetting and dispersing agents) in the commercial micronized sulphur preparation used (‘Thiovit’) were inhibitory at high concentrations but stimulatory at low concentrations. The significance to field conditions is discussed.     

选择性抑菌剂对添加稻草红壤旱土团聚体磷素微生物转化的影响

为了进一步探明红壤旱土磷素微生物固持的机理,采用室内模拟培养试验研究了微生物类群对红壤旱土团聚体(0.2～2mm)磷素转化的作用.结果表明:在培养90d期间,添加稻草处理能显著提高红壤旱土团聚体的微生物生物量碳、生物量磷、提取磷(Olsen法)和有机磷的含量.在培养前期(5～30d),与添加稻草处理比较,稻草+真菌抑制剂(放线菌酮)、稻草+细菌抑制剂(四环素+链霉素硫酸盐)处理团聚体微生物生物量碳含量分别降低10.5%～31.8%和6.8%～11.6%,前者的降低幅度显著大于后者(P0.01),此后加入抑菌剂处理团聚体微生物生物量碳基本保持稳定.添加细菌抑制剂处理团聚体微生物生物量磷含量在培养5～20d期间比加真菌抑制剂处理高10.0%～28.8%,差异显著(P0.01).表明真菌和细菌均参与红壤旱土团聚体磷素的固持,但真菌的作用明显大于细菌.     

The evaluation of plant-available sulphur in soils. II

Summary The availability to oats of adsorbed sulphate in soils and of sulphate impurity in calcium carbonate was studied in pot-culture experiments.When calcium carbonate was added to soils with pH values ranging from 5.7 to 7.4 the uptake of sulphur by oats was increased, due probably to enhanced mineralization of soil organic sulphur. When the calcium carbonate contained sulphate impurity the uptake of sulphur was further increased by an amount comparable with the release of sulphate which could be expected from a reaction of the calcium carbonate with the exchangeable hydrogen of the soil. Sulphate in excess of this amount appeared to be largely unavailable. Uptake of sulphur by oats from calcareous sands containing large amounts of insoluble sulphate associated with calcium carbonate also suggested that soil sulphur in this form had very low availability to plants.Substantial increases in the amounts of sulphur extracted by reagents commonly used for the determination of adsorbed sulphate in soils occurred when soils were airdried at about 20°C. Decreases in adsorbed sulphate in soils following the growth of oats in pot culture confirmed that adsorbed sulphate is readily available to plants.     

Water-soluble carbon in roots of rape and barley: impacts on labile soil organic carbon, arylsulphatase activity and sulphur mineralization

Investigating the impact of plant species on sulphur (S) availability in the rhizosphere soil is agronomically important to optimize S fertilization. Bulk, rhizosphere soils and the roots of field-grown rape and barley were sampled 7 times (every fortnight), from March to June, at plant maturity. Root carbon (C) and nitrogen (N) in water extract, along with soil SO₄²⁻-S, labile soil organic-C (HWC) and -N (HWN) in hot water extract, as well as soil arylsulphatase activity were then monitored. The average concentrations of both HWC and HWN were observed in the following decreasing order: rape rhizosphere soil >barley rhizosphere soil >bulk soil. In parallel, the average contents of water extractable-C and -N in rape roots were higher than those in barley roots. These results suggest that soil C and N contents in hot water extract (including rhizodeposition) were correlated with C and N released by roots. Great ARS activities found in rape rhizosphere soil were accompanied by great SO₄²⁻-S mineralization over time. Finally, bulk and rhizosphere soils of rape and barley were pooled from the seven samplings and incubated with the corresponding pooled root water-soluble C of both plant species and glucose-C. After 1 and 9 weeks, a greater net S mineralization (gross mineralization - immobilization) was observed with rape root water-soluble C than with barley root water-soluble C and glucose-C. Conjointly, we found a higher average value of ARS activity in rape rhizosphere than in barley rhizosphere soil. Our findings suggest that plant species, via their rhizodeposition, determine the dynamic of S in soil.     

模拟酸雨对不同土壤有机碳和作物秸秆分解的影响

为研究酸雨对不同pH值水稻土中有机碳分解的影响,选择酸性（pH 5.48）、碱性（pH 8.18）和中性（pH 6.70）水稻土（分别设置施用秸秆0、15 g·kg^-1土处理）在20 ℃条件下进行40 d的培养试验,各土壤组分别用pH值为6.0、4.5、3.0的模拟雨水将土壤含水量调为400 g·kg^-1(以风干土计).结果表明：秸秆、酸雨和土壤共同对土壤系统CO₂释放产生影响,秸秆的添加可显著提高土壤CO₂释放速率.培养期间,酸雨未显著影响土壤有机碳分解,但对土壤中作物秸秆的分解影响显著. pH 3.0酸雨处理下酸性和碱性土壤中秸秆40 d总分解量比pH 6.0处理高8%;酸雨抑制了中性水稻土中秸秆的分解,pH 3.0酸雨处理下秸秆40 d总分解量比pH 6.0处理低15%.pH 3.0酸雨处理下,酸性水稻土有机碳分解速率分别比中性和碱性水稻土高43%和50%（P<0.05）,秸秆在中性水稻土中分解量分别比其在酸性和碱性水稻土中低17%和16%（P<0.05）.