Evolution of humic substances from unripe compost during incubation with lignolytic or cellulolytic microorganisms and effects on the lettuce growth promotion mediated byAzotobacter chroococcum

Some recalcitrant organic wastes, which contain a large proportion of lignin or cellulose, are not changed much by composting, and thus the effectiveness of the compost as fertilizer is usua.lly low. In this study, incubation of unripe compost with ligno-cellulolytic microorganisms —Trichoderma viride orBacillus spp. — was investigated to increase the degree of humification of the organic matter present, and improve ils quality as a soil amendment. High-performance liquid chromatography (HPLC) analyses together with humification indices and electrofocusing patterns were used to monitor the evolution of the humic substances during the incubation process. Plant growth effects exerted by Azotobacter chroococcum on lettuce plants growing on the previously incubated compost were affected by the length of incubation and by changes in the composition of humic substances. Higher organic matter content and better humification seem to be important factors for predictingA. chroococcum hebaviour in the rhizosphere.     

Composting And Humification

The development of humic substances in the course of four composting processes was monitored quantitatively, recording both relative and absolute contents. Relative data showed contrasting results if the humic substances (HS) were related to the dry matter (d.m.) or to volatile solids (VS). Humic substances were apparently formed because of a concentration effect due to organic matter degradation. If absolute contents were considered, a decrease in the humic substances was observed, above all in the early stages of the process, due, probably, to degradation of the organic material, such as proteins, carbohydrates and lipids, coextracted with the humic substances. Processing of the data in respect of humic substance content over 13 composting processes and one study on the degradation of plant residues in soils, confirmed that no net humic substances are formed during composting and that the humification should be interpreted merely as degradation of the organic matter associated with the humic substances, after uncovering what is known as the core of the humus. The concept of humification during the composting process therefore needs to be reviewed, bearing in mind that neither the method commonly used for humic substance extraction nor the relative results obtained enable it to be interpreted satisfactorily.     

Interaction of heavy metals with the organic matter of an ordinary chernozem

Lead, copper, and zinc interacted with the soil organic matter when applied to an ordinary chernozem in a pot experiment. Two years after the treatment, an appreciable part of the metals applied was found in the organic substances, predominantly in a loosely bound state. These organic substances were supposed to be organomineral complexes, the formation of which resulted in the partial destruction of humic acid molecules. These processes increased the content of aliphatic structures and the share of fulvic acids and decreased the content of humic acids in the organic matter of the chernozem.     

A comparative study of molecular weight distribution of water-soluble humic substances,humic acids,and fulvic acids extracted from sod-podzolic soils

The alteration of organic matter from three sod-podzolic soils in the course of the extraction of water-soluble humic substances was studied by means of size-exclusion chromatography using a combined UV-DOC detector. A comparison of water-soluble humic substances with humic and fulvic acids extracted from the same soils was carried out.     

Abbau von Huminstoffen durch Bodenmikroorganismen — eine Übersicht

Degradation of humic substances by soil microorganisms — a review Humic substances which represent differently extractable fractions of the soil organic matter exert multifarious effects on soil as a site for plant growth and a part of terrestrial environments. Among them especially humic acids and fulvic acids are subject to degradation and/or transformation by soil microorganisms. Several authors demonstrated the participation of different species of fungi, actinomycetes and also of non-mycelial aerobic or anaerobic bacteria in those processes under laboratory conditions. Indications exist that humic substances irrespective of their structure undergo degradation on cell surfaces due to the activity of exoenzymes. In this respect microbial phenoloxidases play an extraordinarily important role. The degradation rate of humic substances can be followed by optical, gravimetric and chemoanalytical methods as well as using biochemical and microbiological procedures (CO₂ release, microbial growth, biomass formation). An objective evaluation, however, can be hindered by the adsorption of humic substances on microbial biomass and sometimes also by formation of novel humic-like microbial metabolites. Therefore it is necessary to apply a multifactorial approach in the study of the degradation of humic substances which includes both quantitative and qualitative parameters. To better elucidate how these processes may occur under natural conditions, mixed populations of soil microorganisms should be predominantly involved in future studies.     

Water- and pyrophosphate-extractable humic substances fractions as a source of iron for Fe-deficient cucumber plants

The capacity of Fe-deficient cucumber plants to utilise water-extractable and pyrophosphate-extractable humic substances as a source of Fe was investigated. Plants were grown for 13 days in nutrient solution in the presence or absence of Fe and during the last 7 days water-extractable and pyrophosphate-extractable humic substances were added to the solution at a final concentration of 5 μg organic C ml^–1. The water-extractable humic fraction did not significantly modify leaf area and dry matter accumulation, leaf total Fe or chlorophyll content of cucumber plants adequately supplied with Fe. In contrast, pyrophosphate-extractable humic substances caused a slight but significant decrease of all the leaf parameters considered, with the exception of the chlorophyll content. Root Fe content of Fe-sufficient plants was decreased by more than 50% in the presence of each humified fraction. Addition of each humic fraction to Fe-deficient plants led to a partial disappearance of leaf chlorosis symptoms with a significant increase in chlorophyll and leaf Fe content. Fe content of roots was also significantly increased in Fe-deficient plants by the addition of humic substances to the nutrient solution. These results show that Fe-deficient cucumber plants can utilise Fe contained in the two fractions of humified organic matter. However, by calculating the amount of total Fe accumulated per plant in the presence of water-extractable or pyrophosphate-extractable humic substances, it could be seen that Fe contained in the water-extractable humic fraction was almost totally used by Fe-deficient cucumber plants, while that present in the pyrophosphate-extractable fraction could only be partially absorbed. The results strongly support a role of humified organic matter in Fe nutrition of plants and are discussed in terms of a possible interaction between soil humic substances and the biochemical mechanisms involved in the plant response to Fe deficiency. Received: 6 November 1996     

The distribution of the stable carbon isotope (13C/12C) in fractions of soil organic matter

The carbon-isotopic composition of fulvic and humic acid from the A horizons of eight soil types, developed under a wide variety of climatological conditions, was measured. The fulvic acid is always enriched in ¹³C as compared with the humic acid from the same soil by a rather constant factor of 0.9?. The fulvic acids are isotopically closer to the plant source of the organic matter and thus represent an intermediate stage in the formation of humic substances. Depth sections of peat soil showed that carbon isotopes can be used to evaluate the dynamic nature of the fulvic-acid fraction. With depth, a transfer of carbon groups from polysaccharides to fulvic acid is seen. Based on isotopic evidence it is shown that in addition to formation of β-humus, part of the fulvic acid is condensed with depth to a stable humic fraction — humin.     

Mikrosondenuntersuchungen an unterschiedlich stark mit Schwermetallen belasteten Böden. 2. Gehalte an Schwermetallen und anderen Elementen in Huminstoffaggregationen,Streustoffen und Holzkohlepartikeln

Electron microprobe studies on soil samples with varying heavy metal contamination. 2. Contents of heavy metals and other elements in aggregations of humic substances, litter residues and charcoal particles EMA point analysis show that the organic matter constituents of heavy metal contaminated soils are highly enriched with heavy metals. The maximal trace element accumulation were for Cu up to 13,000 mg/kg, for Zn up to 48,000 mg/kg, for Cd up to 2,100 mg/kg and for Pb up to 193,000 mg/kg. The affinity for the accumulation of the different heavy metals in aggregations of humic substances can be described by the sequence Cu > Pb ? Cd > Zn ? Ni > Co. In very strongly acidified humic top soil horizons the Pb and Cd accumulation in the organic matter constituents is in competition with the accumulation in Fe and Mn oxides. The heavy metal contents (especially of Cu) of the organic matter are often correlated with the content of organically bound calcium. The EMA results also show that high heavy metal amounts occur in combination with Ca-accumulations in the epidermis and the outer bark parenchym of decayed roots. EMA point analysis of the interior of fungus sclerotias show that sclerotias can contain high amounts of heavy metals, in particular lead (up to 49,700 mg Pb/kg). From statistical results of EMA point analysis follows that lead and other heavy metals attached to humic substances are not only bound as metal organic complexes but also as organic metal phosphate complexes. Also charcoal particles of polluted soils contain high amounts of heay metals. The accumulation affinity is quite similar to that of humic substances.     

Characterization by isoelectric focusing of the organic matter of a regenerated soil

Abstract

Regeneration of a soil with a high degree of desertification was conducted by the addition of different doses of municipal solid waste organic matter (MSW). Five years after this treatment, humic substances were extracted from these soils and characterized by spectroscopy and isoelectric focusing. No significant differences between E4/E6 ratio and ?log K (Log A400 nm‐log A600 nm) were observed for humic substances extracted from treated and untreated soils. However, the isoelectric focusing (IEF) technique established differences between the humic substances from control and treated soils. The focusing pattern of the former showed a well defined band at pH 9.1 which nearly disappeared in the soils with high doses of MSW (1–2 %). However the organic matter which focused at pH 5.8 was present in all soils.     

Effect of tillage and liming on organic matter composition in a Rhodic Ferralsol from Southern Brazil

The effects of tillage and liming on degradation of organic matter of a Rhodic Ferralsol were investigated. Samples were taken from a tillage experiment and from a trial with different levels of lime application. Organic matter was separated into the fractions of undecomposed organic matter and humic substances by density fractionation. Chemical composition of the humic substances was determined by Kononova's fractionation method. Other chemical parameters measured were total organic C, pH, exchangeable cations, and the physical property determined was aggregate stability. Tillage, and to a lesser degree, liming, substantially decreased organic matter content when compared to samples taken from a virgin forest. The proportion of humic acids C (HC) varied most, so that the ratio of fulvic acids C (FC) to HC increased from 2.5 (virgin forest), to 3.4 (lime treatments and no-tillage) and to 5.8 (conventional tillage). Content of FC was affected by pH, while no distinct factors were found to influence HC content. Aggregate stability was best related to content of HC, confirming the importance of this fraction of organic matter for aggregation. It was concluded that especially excessive tillage greatly affects soil fertility because organic matter is decomposed to a great extent. However, a further need for research seems necessary to clarify interactions conclusively between changes of pH, adsorption of humic substances and other chemical properties in the entire profile of Ferralsols.     

Effect of soil invertebrates on the formation of humic substances under laboratory conditions

The complete polymerization of phenols and proteins (one of the processes involved in the formation of humic substances) was explained. It was shown that fly (Bibio marci) larvae and earthworms (Aporrectodea caliginosa) participate in the complete polymerization of phenols and proteins. In a laboratory experiment, invertebrates participated in the degradation of organic matter and the synthesis of humic substances, which was proved in experiments with ¹⁴C-labeled phenols and proteins. The same organic substances (phenols and proteins) without the impact of invertebrates were used as the control substances. The distributions of the ¹⁴C isotope in alkaline extracts separated by solubility in acids (humic and fulvic acids) was compared to those of the control substances. The portion of the ¹⁴C isotope in the humic acids in the excrements of Bibio marci was higher than that in the control substances. The content of ¹⁴C-labeled humic substances in the excrements of the earthworm Aporrectodea caliginosa exceeded the control values only in the experiment with proteins. When clay material was added to the organic substances, the portion of the ¹⁴C isotope in the humic acids increased in both experiments with phenols and proteins. When these substrates passed through the digestive tracts of the invertebrates, the polymerization of organic substances and the inclusion of proteins and phenols into humic acids occurred.     

Interpretation of humic acid coagulation and soluble soil organic matter using a calculated electrostatic potential

The coagulation of humic substances and its role in controlling the solubility of organic matter in soils are not well understood. We therefore studied the physico‐chemical behaviour of purified humic acid from forest soil coagulated with Na, Ca, Cu, Al at pH 4 and 6, and then modelled the behaviour with the Non‐Ideal Consistent Competitive Adsorption Donnan (NICA‐Donnan) model. We found that the coagulation of humic acid occurs when the Donnan potential is less negative than ?0.08 V. Based on this result, an empirical relation between the Donnan potential of humic acid and its concentration in solution was derived. In addition, the Donnan potential of the dissolved organic matter in the soil solution of six soil profiles from forests was calculated using the NICA‐Donnan model under the assumption that all the dissolved organic matter behaves as humic acid. The measured concentration of dissolved organic matter also decreases in a soil profile, as the calculated potential becomes less negative. The results are in many cases in semi‐quantitative agreement with the predicted concentration based on the humic acid coagulation experiment. Acid soils contain more dissolved organic matter, which may result from the presence of a fairly large fraction of more soluble organic molecules, such as fulvic acid.     

Über Veränderungen im Humuskomplex bei der Degradation von Schwarzerden (Tschernosemen)

Dynamics of organic matter during the degradation of Chernozems In the course of pedogenesis especially the amount and the composition of soil organic matter changes. This is investigated in the presented paper at 6 different developmentstages of a Chernozem-degradation-sequence from Hildesheim region. The mechanisms of this process can be deduced from the ‘degradation products’, found in the different soils. Using several methods for extraction of organic substances by successively stronger reagents or conditions, different amounts of fulvic and humic acids were got. The findings give indications to the different stability of bondings between mineral and organic substances in the soils and their change during degradation. The ascending exhaustive fractionating method was found to be the best for obtaining and-simultaneously - separating completely the organic matter in distinct franctions and also to earn a humus-clay and a silica humic acid component. Using this method, 5 fractions of the humus complex at any of profile-horizons were got in greater amounts and quantitative as far as qualitative determined. It was found that the loss of organic matter is differently distributed to the single humus fractions in the profiles and in the horizon-zones. Therefore it is not sufficient to determine only the total C- and N-content. Comparing the increase or decrease of humic substances in the upper resp. lower zones of the profiles of the sequence, the degradation process can be pursued retrospectively.     

Humic materials from an organic soil: A comparison of extractants and of properties of extracts

Five extraction procedures and thirteen extracting reagents, which included dipolar aprotic solvents, organic chelating agents, pyridine, ethylenediamine, sodium hydroxide, ion-exchange resins and two salts (sodium pyrophosphate and ethylenediamine hydrochloride), were used to extract humic materials from an organic soil. Extractabilities increased in the general order: salts < organic chelating agents < dipolar aprotic solvents < pyridine < ethylenediamine = sodium hydroxide, and the amounts of the soil organic matter extracted by the reagents in the series ranged from 13 to 63%. Gel chromatography techniques indicated that extracts in dipolar aprotic solvents were predominantly of intermediate and low molecular-weight values, and it is suggested that the more highly oxidised soil humic materials were extracted in these. The more efficient solvents extracted materials with a range (high—low) of molecular-weight values. Data from elemental analysis and from E.S.R. measurements indicated that ethylenediamine altered the chemical nature and the composition of extracts. Dipolar aprotics, by the same criteria, were found not to alter the humic extracts, and can be regarded as mild reagents for the extraction of a less representative (of the total) fraction of soil organic matter. Sodium hydroxide in solution, despite its oxidation effects, was the best of the reagents tested for isolating extracts which were representative of a wide range of soil humic substances.     

Transformation of organic matter from maize residues into labile and humic fractions of three European soils as revealed by 13C distribution and CPMAS-NMR spectra

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.     

Influences of Forest Decline on Various Properties of Soils at Mt. Hirugatake, Tanzawa Mountains, Kanto District, Japan

At Mt. Hirugatake in the Tanzawa Mountains, Kanto district, Japan, the deciduous broadleaved forests have rapidly declined. In our previous studies, we reported that the amount of soil organic matter had significantly decreased at the early and final stages of forest decline, and that the soil microbial biomass also showed a large decrease at these stages, suggesting that the composition of soil organic matter might have also changed with forest decline. To clarify the influences of forest decline on the composition of soil organic matter, the amount of humic substances, optical properties of humic acid, and the amount of soil carbohydrates in surface soils at different stages of forest decline were investigated. The amounts of humic acid and fulvic acid decreased to a lesser extent at the early and middle stages of forest decline, and showed a significant decrease at the final stage. As the amount of humin significantly decreased at the early stage, it was plausible that the distinct decrease in the total carbon content of the soil surface horizons at the early stage of forest decline was induced by the decrease in the amount of humin, and at the final stage, by the decrease in the amounts of humic acid and fulvic acid. The amount of soil carbohydrates did not change appreciably with forest decline although the soil organic matter content markedly decreased. It was suggested that most of the carbohydrates in the soil surface horizons were in a stabilized form consisting of complexes with humic substances, metals, and minerals, and would not be affected by the environmental changes associated with forest decline.     

Modelling pH buffering and aluminium solubility in European forest soils

The pH buffering and aluminium solubility characteristics of acid soil are important in determining the soil's response to changes in precipitation acidity. The chemistry of soil organic matter (humic substances) plays a key role in both processes, yet is complex and still poorly understood. Nevertheless, models of humic substance chemistry have been developed, one of which is WHAM–S, which contains a model (Model V) of proton and metal binding at discrete sites on humic substances and considers electrostatic effects on the binding strength. Here we have tested the ability of WHAM–S to model solution pH and Al using batch titration studies on organic and mineral soil horizons from forested sites in Norway, Germany and Spain, with ambient pH values from 3.73 to 5.73. We optimized the model predictions by adjusting the amounts of soil aluminium and humic substances within defined limits, taking the contents of copper chloride‐extractable Al and the base‐extractable organic matter as starting values. The model simulated both pH and dissolved Al well with optimized amounts of aluminium and humic substances within the defined limits (root mean squared error for pH from 0.01 to 0.22, for p[Al]_aq (total dissolved Al) from 0.03 to 0.49, five data points). Control of dissolved Al by dissolved organic matter was important particularly at above‐ambient pH. In two mineral horizons we improved the fits by assuming that Al could precipitate as Al(OH)₃. The optimized model also gave reasonable predictions of pH and dissolved Al in supernatants obtained by repeated leaching of the soil horizons. The results show that humic substances dominate the control of pH and dissolved Al in most of the horizons studied. Control by Al(OH)₃ occurs but is the exception.     

Properties of neutral phosphate buffer extractable organic matter in soils revealed using size exclusion chromatography and fractionation with polyvinylpyrrolidone

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.     

Soil Organic Matter Content and Quality in Polyfactorial Field Experiments

The polyfactorial long-term field experiments founded in 1979 exist in four different sites till the present time. The experimental design and selected variants have been described in our previous paper (Lipavsky et al. 2002). Besides of the organic carbon and nitrogen contents in soil, dry matter of the main and second products, nitrogen uptake by the main and second products presented in our previous paper (Lipavsky et al, 2002), hot water soluble carbon (C hwl ), humic substances, e.g. humic and fulvic acids have been determined in soil samples taken in autumn each year from six selected variants of organic and mineral fertilisation. The results of the hot water soluble carbon content and humic and fulvic acids contents in the air dried soil samples have been evaluated in this paper. It was shown that humic substances are rather conservative and site specific soil properties that are not much affected by the cropping and fertilisation systems. Hot water soluble carbon, on the other hand, seems to be a dynamic part of the soil organic matter.     

沈阳市郊玉米连作土壤有机质组成及其对土壤结构的影响

通过对沈阳市郊玉米连作土壤的有机质、结构情况以及二者相互关系研究表明,土壤中有机质大致范围在16.31gkg-1～23.72gkg-1,腐殖质大致范围在15.17gkg-1～21.49gkg-1,非腐殖质大致范围在1.14gkg-1～2.59gkg-1;在腐殖质各组分中,胡敏素的含量最高,约占腐殖质总量的58.93%～66.34%,胡敏酸约占16.26%～22.87%,富里酸约占16.82%～18.20%,胡富比约在0.95～1.32之间;干筛处理后>0.25mm的土壤团聚体占到80%以上,其中以>2mm团聚体为主,湿筛处理后>0.25mm的土壤团聚体仅占20%左右,其中0.25～0.5mm团聚体为主,水稳性系数在17.33%～35.67%。对有机质及其各组分与土壤各级团聚体做的相关分析结果表明,有机质与0.25～5mm粒径组的团聚体呈显著相关,与>5mm团聚体不相关,且胡敏酸主要影响1～5mm大团聚体的形成。