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1.
The basic principles of the electrophoretic behaviour of humic substances (HS) still need to be systematically investigated. HS extracted from a Cryorthod (HS–1) and from an Haplumbrept (HS–2) were first fractionated by ultrafiltration to obtain two series of fractions of reduced polydispersity with a nominal mean relative molecular mass (M) of 7500, 20000, 40000, 75000 and 200 000 Da. Fractions extracted from the same soil behaved like an homologous series when tested by size exclusion chromatography, showing a linear relation of M with molecular size. When plotted against their mean M, the elution volumes of HS fractions extracted from the two different soils originated lines of different slope. Absolute electrophoretic mobilities of the fractions were determined in polyacrylamide gel slabs of increasing polyacrylamide concentration using a single buffer system; in all gels there was a close linear relation between the electrophoretic mobility and the logarithm of the mean M of HS fractions. Extrapolation of mobilities of HS–1 at zero gel concentration gave intercepts that did not differ significantly, showing that there was a constant mobility for all fractions in free solution. These results mean that charge differences have little effect on the electrophoretic mobilities of HS extracted from the same soil and imply the theoretical possibility of determining M distributions of HS by polyacrylamide gel electrophoresis in a single buffer system. Mobilities of HS–2 fractions were compared with those of HS–1: the latter fitted the regression equations of mobility against logarithm of the molecular weight obtained from HS–1 humic substances only in gels of small acrylamide concentration. Deviations were larger at small M, probably because of the increasing fulvic character of the fractions, and increased in gels of greater acrylamide concentration, indicating that charge differences may not be negligible when comparing humic substances extracted from different soils.  相似文献   

2.
Humic acids (HAs) from four soils were fractionated by size exclusion chromatography (SEC) on Sephadex G-75. Three fractions were obtained in all humic acids, collected and assayed by Polyacrylamide gel electrophoresis (PAGE). The unfractionated HA from each soil was used as reference. Each chromatographic fraction formed one electrophoretic zone corresponding closely to one band of the reference sample with some admixture of the fractions preceding or following. The results indicate that fractionation of HAs by tandem SEC-PAGE can be successfully used for obtaining fractions of reduced polydispersity and different electrophoretic mobilities. Pyrolysis/methylation-gas chromatography-mass spectrometry of the full size preparations of HA and fractions with exactly defined molecular size and electrophoretic mobility showed a different distribution in humic components, mainly lipids, lignin derivatives and N-containing compounds.  相似文献   

3.
 Humic substances are the most dynamic component of agricultural soils. In this study, humic substances were extracted from soils based on standard acid/base solubility and by gel filtration using Sephadex G25. Organic P is a component of humic substances, and inositol phosphates are considered to have high prominence in organic P. The objectives of the study were to determine the effects of municipal solid waste (MSW) compost on the amount of inositol phosphates (IP) present in soils, and compare methods of extracting humic substances from soils. Total IP (IPT) in soil extracts was determined following separation using a AG-1× 8 anion exchange resin, acid digestion and ICAP analysis for P. The percentage of IP (%IP) of the total P in the soil was also determined by colorimetric analysis. The soils were part of an experiment to study the effects of three rates of MSW compost and fertilizers on three crops grown in a Pugwash sandy loam (Humo-Ferric Podzol). The data were compared using ANOVA atP≤0.05; extraction methods, fractions [humic/fulvic, high/low molecular (MW)], and five treatments (three different rates of compost, fertilizer, and untreated soil) were compared. The gel filtration method extracted significantly more IPT and %IP than the solubility method. In the solubility method, humic and fulvic acids contained the same amounts of IPT and %IP, while the high MW fraction always contained more IPT and %IP than the low MW fraction in the gel filtration method. Fulvic acids and the low MW fraction contained similar amounts of IPT and %IP. There were differences in %IP between compost-treated soils and the non-compost-treated soils, although there were no differences in IPT due to rate of compost addition. Received: 20 October 1999  相似文献   

4.
The dynamics of incorporation of fresh organic residues into the various fractions of soil organic matter have yet to be clarified in terms of chemical structures and mechanisms involved. We studied by 13C‐dilution analysis and CPMAS‐13C‐NMR spectroscopy the distribution of organic carbon from mixed or mulched maize residues into specific defined fractions such as carbohydrates and humic fractions isolated by selective extractants in a year‐long incubation of three European soils. The contents of carbohydrates in soil particle size fractions and relative δ13C values showed no retention of carbohydrates from maize but rather decomposition of those from native organic matter in the soil. By contrast, CPMAS‐13C‐NMR spectra of humic (HA) and fulvic acids (FA) extracted by alkaline solution generally indicated the transfer of maize C (mostly carbohydrates and peptides) into humic materials, whereas spectra of organic matter extracted with an acetone solution (HE) indicated solubilization of an aliphatic‐rich, hydrophobic fraction that seemed not to contain any C from maize. The abundance of 13C showed that all humic fractions behaved as a sink for C from maize residues but the FA fraction was related to the turnover of fresh organic matter more than the HA. Removal of hydrophobic components from incubated soils by acetone solution allowed a subsequent extraction of HA and, especially, FA still containing much C from maize. The combination of isotopic measurements and NMR spectra indicated that while hydrophilic compounds from maize were retained in HA and FA, hydrophobic components in the HE fraction had chemical features similar to those of humin. Our results show that the organic compounds released in soils by mineralization of fresh plant residues are stored mainly in the hydrophilic fraction of humic substances which are, in turn, stabilized against microbial degradation by the most hydrophobic humic matter. Our findings suggest that native soil humic substances contribute to the accumulation of new organic matter in soils.  相似文献   

5.
Abstract

Comparisons were made between the chemical compositions of humic substances extracted from three soils covered by different vegetation and their biological activities assayed using 15‐ and 30‐day‐old seedlings of Pinus sylvestris and Picea abies. The growth, “α‐amylase and invertase activity were affected by humic fractions and by gibberellic acid (GA), indicating that humic matter had a gibberellin‐like activity. The isoenzymatic polymorphism in the electrophoretic patterns of esterase was influenced in a similar way by all humic fractions and by indoleacetic acid (IAA), which also suggested that the humic fractions exhibited an auxin‐like activity. The humic fractions extracted from the grassland, exhibiting higher amounts of phenolic and a considerable amount of carboxyl carbon, showed the best metabolic effect. The forest humic fractions, characterized by lower phenolic carbon content, appeared less effective in influencing plant metabolism, whereas the grassland‐forest humic substances proved to be even more less effective. The auxin‐ and the gibberellin‐like activities were related to a high content of phenolic and carboxylic groups. These results presented evidence that the biological activity of the humic substances was attributed to their chemical structure and to their functional groups, which could interact with hormone‐binding proteins in the membrane systems, evoking a hormone‐like response.  相似文献   

6.
The effect of a low molecular weight (<5kDa, LMW) and a high molecular weight (>5kDa, HMW) humic fractions on surface redox activities of oat roots was studied. Oxidation of the electron donor NADH and reduction of the artificial electron acceptor ferricyanide [K3Fe(CN)6] exogenously supplied to the roots both alone or in combination, was measured in the presence or absence of soil humic substances. HMW humic fraction inhibited NADH oxidation either in the presence or absence of ferricyanide, while LMW humic fraction inhibited NADH: ferricyanide oxidoreductase activity due to the contemporary addition of the two redox compounds to the solution bathing the roots. NADH: ferricyanide oxido‐reduction was partially due to the release of substances from the roots. However, the presence of soil humic fractions (LMW or HMW) did not significantly modify this behaviour. Rather, the inhibitory effect of soil humic substances was even more evident when the oxidoreduction solely due to the root activity was considered. The results confirm the presence of multiple types of oxidoreductase activities at the surface of oat roots and show that the two humic fractions HMW and LMW may differently affect these activities. Partial inhibition of NADH oxidase activity is interpreted as a possible way of interference of humic substances with metabolic processes involved in cell wall formation. The effects on surface redox activities are discussed in terms of the role of soil humic substances in promotion of plant growth.  相似文献   

7.
Studies were made to determine the rate of decomposition of some 14C-labeled microbial polysaccharides, microbial cells, glucose, cellulose and wheat straw in soil, the distribution of the residual 14C in various humic fractions and the influence of the microbial products on the decomposition of plant residues in soil. During 16 weeks from 32 to 86 per cent of the C of added bacterial polysaccharides had evolved as 14CO2. Chromobacterium violaceum polysaccharide was most resistant and Leuconostoc dextranicus polysaccharide least resistant. In general the polysaccharides, microbial cells, and glucose exerted little effect on the decomposition of the plant products. Upon incubation the 14C-activity was quickly distributed in the humic. fulvic and extracted soil fractions. The pattern of distribution depended upon the amendment and the degree of decomposition. The distribution was most uniform in the highly decomposed amendments. After 16 weeks the bulk of the residual activity from Azotobacter indicus polysaccharide remained in the NaOH extracted soil. From C. violaceum polysaccharide both the extracted soil and the humic acid fraction contained high activity. About 50–80 per cent of the residual activity from the 14C-glucose, cellulose and wheat straw amended soils could be removed by hydrolysis with 6 n HCl. The greater part of this activity in the humic acid fraction was associated with the amino acids and that from the fulvic acids and residual soils after NaOH extraction with the carbohydrates. About 8 16 per cent of the activity of the humic acid fraction was present in substances (probably aromatic) extracted by ether after reductive or oxidative degradation.  相似文献   

8.
Abstract

Soils contaminated with heavy metals constitute a serious and widespread ecological problem but to clean such soils requires strong chemicals such as polycarboxylates; frequently ethylenediaminetetraacetic acid and nitrilotriacetic acid are used. However, these compounds are synthetic and toxic and their replacement by natural products such as soluble humic substances as washing agents for cleaning heavy metal polluted soils would be environmentally very attractive. In fact, such a replacement seems possible at least on cadmium and copper contaminated soil inasmuch as humic substances, depending on the concentration, were found to extract up to 45% and 54% of total cadmium and copper from a highly contaminated calcareous soil. Even though higher amounts of the two metals were extracted by ethylenediaminetetraacetic acid and nitrilotriacetic acid, the humic substances undoubtedly extracted the most reactive fractions. However, the humic substances extracted only 4% of total lead and 17% of total nickel, whereas the percentages for the synthetic polycarboxylates were about 30% for nickel and lead. Ethylenediaminetetraacetic acid and nitrilotriacetic acid may therefore be replaced by humic substances as washing agents for cadmium, copper and maybe nickel contaminated soils, whereas they seem unsuited for cleaning lead contaminated soils, at least if the soils are as calcareous as the soil tested.  相似文献   

9.
The chemistry of soil organic nitrogen: a review   总被引：6，自引：0，他引：6  
1. From the data presented herein it is possible to deduce the following distribution of total N in humic substances and soils: proteinaceous materials (proteins, peptides, and amino acids) – ca. 40%; amino sugars – 5–6%; heterocyclic N compounds (including purines and pyrimidines) – ca. 35%; NH3–19%; approximately 1/4 of the NH3 is fixed NH4 +. Thus, proteinaceous materials and heterocyclics appear to be major soil N components. 2. Natural 15N abundance levels in soils and humic materials are so low that direct analysis by 15N NMR is very difficult or impossible. To overcome this difficulty, the soil or humic material is incubated with 15N-enriched fertilizer. Even incubation in the laboratory for up to 630 days does not produce the same types of 15N compounds that are formed in soils and humic materials over hundreds or thousands of years. For example, very few 15N-labelled heterocyclics are detected by 15N NMR. Does this mean that heterocyclics are not present? Or are the heterocyclics that are present not labelled under these experimental conditions and therefore not detected by the 15N NMR spectrometer ? Another possibility is that a large number of N heterocyclics occur in soils, but each type occurs in very low concentrations. Until the sensitivity is improved, 15N NMR will not provide results that can be compared with data obtained from the same soil and humic material samples by chemical methods and mass spectroscopy. 3. What is most important with respect to agricultural is that all major N forms in soils are available to organisms and are sources of NH3 or NH4 + for plant roots and microbes. Naturally, some of the NH3 will enter the N cycle. 4. From chemical and pyrolysis-mass spectrometric analyses it appears that N heterocylics are significant components of the SOM, rather than degradation products of other molecules due to pyrolysis. The arguments in favor of N heterocyclics as genuine SOM components are the following: a) Some N-heterocyclics originate from biological precursors of SOM, such as proteinaceous materials, carbohydrates, chlorophyll, nucleic acids, and alkaloids, which enter the soil system as plant residues or remains of animals. b) In aquatic humic substances and dissolved organic matter (DOM) at considerably lower pyrolysis temperatures (200 to 300°C), free and substituted N-heterocyclics such as pyrroles, pyrrolidines, pyridines, pyranes, and pyrazoles, have been identified by analytical pyrolysis (Schulten et al 1997b). c) Their presence in humic substances and soils was also detected without pyrolysis by gel chromatography – GC/MS after reductive acetylation (Schnitzer and Spiteller 1986), by X-ray photoelectron spectroscopy (Patience et al. 1992), and also by spectroscopic, chromatographic, chemical, and isotopic methods (Ikan et al. 1992). 5. While we can see light at the end of the tunnel as far as soil-N is concerned, further research is needed to identify additional N-containing compounds such as N- heterocyclics, to determine whether these are present in the soil or humic materials in the form in which they were identified or whether they originate from more complex structures. If the latter is correct, then we need to isolate these complex N-molecules and attempt to identify them.  相似文献   

10.
It has been shown that the distribution features of the hydrophilic and hydrophobic fractions of humic substances (HSs) in the upper genetic horizons of undisturbed podzolic soils and litho-barrier podzols are similar. The concentrations of the separate HS fractions in the mineral horizons are largely determined by the soil mineralogy. In forest litters of secondary phytocenoses developed after the cutting of spruce forests, an increase in the content of hydrophilic compounds has been observed on loamy deposits at a decrease in the portion of lignin-like organic compounds. An increase in the portion of Al-Fe-humus compounds has been revealed in the mineral soil horizons of secondary phytocenoses. It has been proposed to use the degree of hydrophilicity (D H ) for characterizing the features of the organic matter in taiga soils. An increase in the content of the fractions of occluded organic matter and that bound to minerals (density of 1.6–2.2 g/cm3) has been observed in the eluvial horizons of a young cutover area.  相似文献   

11.
Tandem size-exclusion chromatography and polyacrylamide gel electrophoresis were used for obtaining stable fractions of different molecular sizes and electrophoretic mobilities from chernozem humic acids (HAs). The obtained fractions were analyzed using solid-phase 13C NMR. The tendencies of the changes in the aromatic and aliphatic components of the HA fractions with different molecular sizes and electrophoretic mobilities were experimentally revealed. The aromatic-to-aliphatic carbon ratio Carom (165–108 ppm)/Caliph (108-0 ppm) was used for comparing the degrees of aliphaticity and aromaticity of the HA macromolecules. This ratio increased by more than five times when going from the high-molecular-weight to the low-molecular-weight fractions and largely determined their hydrophilic properties. The obtained results can be useful for the interpretation of the structural organization and the ecological functions of soil HAs and their fractions.  相似文献   

12.
We studied the acid‐base properties of 16 fulvic acids and 16 humic acids isolated from the surface (3–15 cm) and subsurface (> 45 cm) horizons of two types of acid forest soils, derived respectively from amphibolite and granite rocks, under five different types of vegetation. The observed differences between the contents of humic substances in the two types of soils were related to the degree of Al‐saturation of the soil organic matter, as indicated by the molar ratio between pyrophosphate extractable Al and C. Humic fractions were characterized in terms of elemental composition, and CPMAS 13C NMR spectrometry. The contents of carboxylic and phenolic groups were estimated by potentiometric titrations conducted in 0.1 m KNO3 in a nitrogen atmosphere. The fulvic acids contained more carboxylic groups but less phenolic groups than the humic acids: the ratio of phenolic to carboxylic groups in the humic acids was 0.48 ± 0.10 and in the fulvic acids 0.23 ± 0.05. The mean values of the protonation constants of each of the humic substance fractions can be used as generic parameters for describing the proton binding properties. The fulvic acids isolated from the subsurface horizon of the soil contained between 2.6 and 23% more carboxylic groups, and the humic acids between 8 and 43% more carboxylic groups than those isolated from the surface horizon of the same soil.  相似文献   

13.
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14.
The effect of dissolved organic substances on the genesis and functioning of gley-podzolic soils in the lower reaches of the Northern Dvina River (Arkhangelsk oblast) has been studied. It is shown that polyphenols and low-molecular (<1000 a.u.) fractions of fulvic acids bound with Fe (II, III) and Mn (II) ions are present in the ELg and Bfg horizons of these soils and participate in alternating downward and upward migration processes. The presence of these compounds colors the soil mass into the dark gray color resembling the color of the proper humus horizon.  相似文献   

15.
Soil organic matter was extracted by a mixture of O.IM Na4P2O: O.IM NaOH from a chronosequence of weakly weathered soils developed on aeolian sand, and fractionated into humin (non-extractable), humic acid, and fulvic acid. The mass of total organic carbon in the profiles, the 14C content and the 13C/12C ratios were also determined. The weight of total carbon increased rapidly at first and then gradually without attaining a steady state. This trend was also shown by the humin and fulvic acid fractions, but the humic acid fraction appeared to have reached a maximum after about 3000 years. The order of total weights of the organic fractions was humin > fulvic acid > humic acid. The evidence suggests that the proportions of the humic fractions formed by decomposition are related to soil differences but not to vegetation. The greater part of the plant material found in the soils appears in the humin and fulvic acid fractions.  相似文献   

16.
Microbial communities are responsible for soil organic matter cycling and thus for maintaining soil fertility. A typical Orthic Luvisol was freed from organic carbon by thermal destruction at 600°C. Then the degradation and humification of 14C‐labelled maize straw by defined microbial communities was analysed. To study the role of microbial diversity on the humification of plant material, microcosms containing sterilized soil were inoculated with a natural microbial community or with microbial consortia consisting of bacterial and fungal soil isolates. Within 6 weeks, 41 ± 4% of applied 14C‐labelled maize straw was mineralized in the soil microcosms containing complex communities derived from a soil suspension, whilst the most efficient communities composed of soil isolates mineralized less than 35%. The humification products were analysed by solution state 13C‐NMR‐spectroscopy and gel permeation chromatography (GPC). The analyses of humic acids extracts by solution state 13C‐NMR‐spectroscopy revealed no difference in the development of typical chemical functional groups for humic substances during incubation. However, the increase in specific molecular size fractions of the extracted humic acids occurred only after inoculation with complex communities, but not with defined isolates. While it seems to be true that redundancy in soil microbial communities contributes to the resilience of soils, specific soil functions may no longer be performed if a microbial community is harshly affected in its diversity or growth conditions.  相似文献   

17.
X－ray photoelectron spectroscopy(XPS) was applied to examine the N structures of soil humic substances and some of their analogues.It was found that for soil humic substances XPS method gave similar results as those obtained by ^15N CPMAS NMR (cross-polarization magic-angle spinning nuclear magnetic resonance) method.70%-86% of total N in soil humic substances was in the form of amide,and 6%-13% was presented as ammes,with the remaining part as heterocyclic N.There was no difference in the distribution of the forms of N between the humic substances from soils formed over hundreds or thousands of years and the newly formed ones.For fulvic acid from weathered coal and benzoquinone-(NH4)2SO4 polymer the XPS results deviated significantly from the ^15N CPMAS NMR data.  相似文献   

18.
Abstract

A neutral phosphate buffer (NPB) extraction method has been used to estimate the amount of available N in soil. However, the properties of soil NPB-extractable organic N have not been fully elucidated. The purpose of the present study was to characterize the properties of organic matter in the NPB extracts of soils. The NPB extracts were obtained from three soil samples, and the organic matter in the extracts was separated into three fractions according to its solubility in acid and adsorption onto polyvinylpyrrolidone (PVP). High-performance size exclusion chromatography (HPSEC) with ultraviolet (UV) and fluorescence detections was applied to the NPB extracts and their fractions. The HPSEC analysis of the NPB extract revealed the presence of a single broad peak, irrespective of the detection methods. The broad peak was identified as humic substances using the on-flow measurements of UV absorption spectra and fluorescence emission spectra. Among the fractions, the PVP-non-adsorbed fulvic acid (FA) fraction accounted for the largest proportion of organic C or N in the NPB extract, followed by the PVP-adsorbed FA and humic acid (HA) fractions. The peak of humic substances was observed for all fractions using HPSEC with the on-flow measurement of UV absorption and fluorescence emission spectra. The molecular weight of the humic substances varied with each fraction. When the Coomassie Blue-reactive substances (CBRS) were quantified using a Bradford protein assay, they were detected in the NPB extract and almost half were distributed in the PVP-non-adsorbed FA fraction. However, humic substances were considered to be the main constituents of CBRS in the soil NPB extract because of their reactivity with Coomassie Blue and the absence of proteinaceous materials. Furthermore, an incubation experiment revealed that the organic matter available to microorganisms was included in the HA and PVP-non-adsorbed FA fractions. Based on the HPSEC analysis of the NPB extracts and their fractions, it was observed that the humic substances in the NPB extract, particularly in the HA and PVP-non-adsorbed FA fractions, were available to microorganisms.  相似文献   

19.
The effect of endogeic earthworms (Octolasion tyrtaeum) and the availability of clay (Montmorillonite) on the mobilization and stabilization of uniformly 14C-labelled catechol mixed into arable and forest soil was investigated in a short- and a long-term microcosm experiment. By using arable and forest soil the effect of earthworms and clay in soils differing in the saturation of the mineral matrix with organic matter was investigated. In the short-term experiment microcosms were destructively sampled when the soil had been transformed into casts. In the long-term experiment earthworm casts produced during 7 days and non-processed soil were incubated for three further months. Production of CO2 and 14CO2 were measured at regular intervals. Accumulation of 14C in humic fractions (DOM, fulvic acids, humic acids and humin) of the casts and the non-processed soil and incorporation of 14C into earthworm tissue were determined.Incorporation of 14C into earthworm tissue was low, with 0.1 and 0.44% recovered in the short- and long-term experiment, respectively, suggesting that endogeic earthworms preferentially assimilate non-phenolic soil carbon. Cumulative production of CO2-C was significantly increased in casts produced from the arable soil, but lower in casts produced from the forest soil; generally, the production of CO2-C was higher in forest than in arable soil. Both soils differed in the pattern of 14CO2-C production; initially it was higher in the forest soil than in the arable soil, whereas later the opposite was true. Octolasion tyrtaeum did not affect 14CO2-C production in the forest soil, but increased it in the arable soil early in the experiment; clay counteracted this effect. Clay and O. tyrtaeum did not affect integration of 14C into humic fractions of the forest soil. In contrast, in the arable soil O. tyrtaeum increased the amount of 14C in the labile fractions, whereas clay increased it in the humin fraction.The results indicate that endogeic earthworms increase microbial activity and thus mineralization of phenolic compounds, whereas clay decreases it presumably by binding phenolic compounds to clay particles when passing through the earthworm gut. Endogeic earthworms and clay are only of minor importance for the fate of catechol in soils with high organic matter, clay and microbial biomass concentrations, but in contrast affect the fate of phenolic compounds in low clay soils.  相似文献   

20.
A humic acid (HA) isolated from a volcanic soil was separated in three fractions of decreasing molecular size (I, II and III) by preparative high performance size exclusion chromatography (HPSEC). The molecular content of the bulk soil HA and its size fractions was characterized by pyrolysis-GC-MS (thermochemolysis with tetramethylammonium hydroxide) and NMR spectroscopy. All soil humic materials were used to evaluate their effects on the enzymatic activities involved in glycolytic and respiratory processes of Zea mays (L.) seedlings. The elementary analyses and NMR spectra of the humic fractions indicated that the content of polar carbons (mainly carbohydrates) increased with decreasing molecular size of separated fractions. The products evolved by on-line thermochemolysis showed that the smallest size fraction (Fraction III) with the least rigid molecular conformation among the humic samples had the lowest content of lignin moieties and the largest amount of other non-lignin aromatic compounds. The bulk HA and the three humic fractions affected the enzyme activities related to glycolysis and tricarboxylic acid cycle (TCA) in different ways depending on molecular size, molecular characteristics and concentrations. The overall effectiveness of the four fractions in promoting the metabolic pathways was in the order: III>HA>II>I. The largest effect of Fraction III, either alone or incorporated into the bulk HA, was attributed to a flexible conformational structure that promoted a more efficient diffusion of bioactive humic components to maize cells. A better knowledge of the relationship between molecular structure of soil humic matter and plant activity may be of practical interest in increasing carbon fixation in plants and redirect atmospheric CO2 into bio-fuel resources.  相似文献   

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