Acid properties of fulvic and humic acids isolated from two acid forest soils under different vegetation cover and soil depth

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.     

The Effect of Severe Drought on the Evolution of Urban and Manure Wastes in an Agricultural Soil in Mediterranean Ecosystems

In semi‐arid Mediterranean soils, water availability is the most limiting factor, negatively affecting the organic matter (OM) degradation. The aim of this work is to study under controlled laboratory conditions how three sources of OM [municipal solid waste (MSW), sheep manure (SM) and cow manure (CM)] behave when they are applied to an agricultural soil subjected to a severe year‐long drought. In order to apply the same concentration of OM to the soil (16·92 Mg OM ha⁻¹), 2 kg of soil was mixed with 30, 67·41 and 55·25 Mg ha⁻¹ (dry matter) of MSW, CM and SM, respectively. Two levels of irrigation were employed: (i) watered soils and (ii) non‐watered soils. Soil's chemical properties [water soluble carbon (WSC), humic acids, fulvic acids and protein mass distribution], biological properties (soil microbial biomass carbon and o‐diphenoloxidase activity) and solid‐state ¹³C cross‐polarisation magic angle spinning nuclear magnetic resonance spectroscopy were determined. In watered soils, the soil microbial biomass carbon was higher in the SM than in CM and MSW treatments (9·9% and 23·1%, respectively). The WSC was significantly higher in SM than in CM (55·7%) and MSW (78·7%) treatments. A decrease in the content of O‐alkyl C and an increase in alkyl C, aromatic C and carboxyl C were observed. In non‐watered soils, the biochemical properties and alkyl C and alkyl/O‐alkyl ratio decreased, whereas WSC content and O‐alkyl C increased. These results indicated that the evolution of OM and the activity of the microbial community in non‐watered soils were very different to those in the watered soils. Copyright © 2016 John Wiley & Sons, Ltd.     

Humus quantity and quality of an entic Haplustoll under different soil‐crop management systems

Abstract

The effects of different management systems on the level and composition of humified organic matter in an entic Haplustoll from the semiarid Pampean region were studied. The systems were: TPc, wheat‐mixed pasture; TV, wheat (Triticum aestivum), oat (Avena sativa), corn (Zea mays) and triticale grasses; TP, wheat‐cattle grazing; and V, virgin, non cultivated. Humic acids were extracted, fractionated, and analyzed for their organic carbon (OC) content, elemental composition, and E4:E6 spectral ratios. The infrared (IR), electron spin resonance (ESR). and ¹³C‐NMR spectra were registered on these humic acids. The TP rotation showed the lowest humic acid‐carbon to fulvic acid‐carbon (HA‐C:FA‐C) ratio. The lower O:C ratio of humic acids from the cropped soils indicates a higher level of oxidation than that of the virgin one. The comparison of the different methodologies allowed us to conclude that crop rotations and conservation tillage were adequate to mantain the level and composition of the soil organic matter and humus which affected the soil fertility and level of productivity     

Analysis of the variable charge of two organic soils by means of the NICA-Donnan model

We have tested to see if the generic set of NICA‐Donnan model parameters, used to describe isolated humic substances, can also describe soil humic substances in situ. A potentiometric back‐titration technique was used to determine the variable surface charge of two organic peat soils at three different ionic strengths. The non‐ideal, competitive‐adsorption NICA‐Donnan model was used to simulate the surface charge, by assuming a bimodal distribution of H⁺ affinity on the soil solid phase. The model provided an excellent fit to the experimental data. The Donnan volume, V_D, varied slightly with ionic strength, although the variation was less than for humic substances in solution. The values obtained for the parameters that define the affinity distributions, the intrinsic proton binding constant (log K_i^int) and the heterogeneity of the site (m_i), were similar to those observed for isolated soil humic acids. The abundance of carboxylic groups in the whole soil represented 30% of the typical value for isolated soil humic acids. The composition of the organic matter of the whole soils, obtained by ¹³C CPMAS NMR, was comparable to the characteristic composition of soil humic acids.     

The influence of humus fractionation on the chemical composition of soil organic matter studied by solid-state 13C NMR

Four samples of soil organic matter and their humic acids, fulvic acids and humin were studied with solid-state ¹³CP MAS NMR. The whole soil samples were fractionated using NaOH and HCl in order to extract humic acids, fulvic acids and humin. This investigation indicates that conventional humus fractionation does not significantly change the content of different functional groups in soil.     

Chromatographische Unterscheidung von Huminstoffen und Nichthuminstoffen aus Schwarzerdehumus

Chromatographic distinction of humic and non humic substances in chernozem-humus Fulvic acids, brown humic acids and grey humic acids from a chernozem from the Austrian Marchfeld were analyzed chromatographically on controlled pore glass with pore diameters of 177 and 259 Å. Concentration of coloured material and concentration of carbon were recorded in the chromatogramms. Specific extinctions at 400 nm were calculated for unit of C and maximal values were taken as constants specific for “pure” humic substances. These were 445,6 ± 10,5,388,7 ± 19,3 and 128,9 ± 0,8 for “pure” grey humic acids, brown humic acids and fulvic acids, respectively. With the aid of these constants, “pure” humic substances could be distinguished from accompanying non humic substances. “Pure” humic substances amounted to 63%, non humic substances to 37% of extractable organic C. 53%, 22% and 25% of the carbon of “pure” humic substances were from grey humic acids, brown humic acids and fulvic acids, respectively.     

Effect of continuous compost application on humus composition and nitrogen fertility of soils in a field subjected to double cropping

We investigated the effect of continuous compost application on humus composition and N fertility of soils in a field subjected to double cropping (paddy rice and barley) for 25 years. Soil samples were collected from three different plots: (a) No-NF, fertilizer containing P and K but no N; (b) F, fertilizer containing N, P, and K; and (c) F+C, fertilizer plus compost. The amounts of total humus, extracted humus, and humic and fulvic acids increased in the order No-NF<F≪F+C. The amounts of humic and fulvic acids were 2.7 and 1.7 times larger in the F+C plot than in the F plot, respectively. The degree of humification of the humic acids decreased in the order No-NF<F<F+C. The absorption curves and ¹³C-NMR spectra (TOSS method) of the humic acids indicated the presence of lignin-like structure, and its degree was the strongest in the F+C plot. The ¹³C-NMR spectra showed distinct differences in the distribution of carbon species between humic and fulvic acids. In humic acids, the content of aromatic-C, ranging from 37 to 44%, was the highest among carbon species. In fulvic acids, the content of O-alkyl-C, ranging from 45 to 51%, was the highest. The amounts of phosphate buffer-extractable N (PEON) and total N (TN) increased in the order No-NF<F<F+C. The amounts of PEON and TN were 1.2 and 1.7 times larger in the F+C plot than in the F plot, respectively. Present and previous findings indicated that continuous compost application could improve various properties of soils in a field subjected to long-term double cropping.     

Reduction of Cr(VI) by soil humic acids

The rate of hexavalent chromium reduction by a soil humic acid (SHA) was investigated in aqueous solutions where concentrations of Cr(VI), H⁺, and SHA were independently varied. Rate experiments were done with a large excess of SHA over Cr(VI). Rates of reduction depend strongly on [H⁺], increasing with decreasing pH. Typical Cr(VI)-SHA reactions display a nonlinear reduction of Cr(VI) with time that cannot be modelled with simple first- or second-order rate equations. An empirical rate equation is developed for Cr(VI)-soil humic acid reactions over a range of experimental conditions. The model is in part based on a reactive continuum concept developed for soil fulvic acids. The rate equation describing Cr(VI) reduction by SHA is: R= -(k₀+k[H⁺]^1/2)[HCrO₄^?]^1/2X_e^?1, where k₀ is (8·3 ± 1·2) × 10^?12, s^?1k is (2·04 ± 0·05) × 10^?9 l^1/2 mol^?1/2 s^?1, and X_e is the equivalent fraction of SHA oxidized. The rate equation adequately models Cr(VI) reduction in an experiment with [Cr(VI)]₀ four times greater than the maximum concentration used in its derivation. Cr(VI) reduction at pH 3 by two other SHAs can also be modelled using the rate equation. The difference between the rate coefficients for the humic acid and the fulvic acid from the same soil was greater than the difference in the rate coefficients for humic acids from different soils.     

Sequential exhaustive extraction of a Mollisol soil, and characterizations of humic components, including humin, by solid and solution state NMR

A comprehensive sequential extraction procedure was applied to isolate soil organic components using aqueous solvents at different pH values, base plus urea (base‐urea), and finally dimethylsulfoxide (DMSO) plus concentrated H₂SO₄ (DMSO‐acid) for the humin‐enriched clay separates. The extracts from base‐urea and DMSO‐acid would be regarded as ‘humin’ in the classical definitions. The fractions isolated from aqueous base, base‐urea and DMSO‐acid were characterized by solid and solution state NMR spectroscopy. The base‐urea solvent system isolated ca. 10% (by mass) additional humic substances. The combined base‐urea and DMSO‐acid solvents isolated ca. 93% of total organic carbon from the humin‐enriched fine clay fraction (<2 μm). Characterization of the humic fractions by solid‐state NMR spectroscopy showed that oxidized char materials were concentrated in humic acids isolated at pH 7, and in the base‐urea extract. Lignin‐derived materials were in considerable abundance in the humic acids isolated at pH 12.6. Only very small amounts of char‐derived structures were contained in the fulvic acids and fulvic acids‐like material isolated from the base‐urea solvent. After extraction with base‐urea, the 0.5 m NaOH extract from the humin‐enriched clay was predominantly composed of aliphatic hydrocarbon groups, and with lesser amounts of aromatic carbon (probably including some char material), and carbohydrates and peptides. From the combination of solid and solution‐state NMR spectroscopy, it is clear that the major components of humin materials, from the DMSO‐acid solvent, after the exhaustive extraction sequence, were composed of microbial and plant derived components, mainly long‐chain aliphatic species (including fatty acids/ester, waxes, lipids and cuticular material), carbohydrate, peptides/proteins, lignin derivatives, lipoprotein and peptidoglycan (major structural components in bacteria cell walls). Black carbon or char materials were enriched in humic acids isolated at pH 7 and humic acids‐like material isolated in the base‐urea medium, indicating that urea can liberate char‐derived material hydrogen bonded or trapped within the humin matrix.     

Quantification of Humic and Fulvic Acids,Macro- and MicroNutrients and C/N Ratio in Organic Fertilizers

Humic substances are the main responsible for soil conditioning and are one of the major components of the composting product. However, there is still little information about their composition and mechanisms of activity. This study aimed to evaluate quantitatively the humic and fulvic fraction by size-exclusion liquid chromatography and the metal content by atomic absorption spectrometry. The chromatographic method used was efficient for humic and fulvic acids quantitation. Levels of 1.73 ± 0.03 and 1.6 ± 0.5 g kg^?1 were found for chicken manure regarding humic and fulvic acids, respectively, and 5.1 ± 0.9 and 1.2 ± 0.1 g kg^?1 for peat. The metal contents indicated the need of a mineral enrichment, because only the levels of iron reached the minimum recommended by legislation. It was also observed that evaluated organic fertilizers were in accordance with the specifications established by legislation in relation to carbon and total nitrogen and the pathogen levels.     

Chemical heterogeneity of humic substances: characterization of size fractions obtained by hollow-fibre ultrafiltration

To investigate the chemical heterogeneity of humic substances in relation to molecular size, fulvic and humic acids were extracted and purified from the surface horizon of a Humic Gleysol in northern Switzerland. A fractionation scheme using hollow‐fibre ultrafiltration cartridges was developed and used to obtain four size fractions of the humic acid with nominal molecular weight ranges > 300 kDa, 100–300 kDa, 30–100 kDa, and 10–30 kDa. The fulvic acid and all humic acid fractions were characterized by size exclusion chromatography, elemental analysis (C, H, N, S), as well as spectroscopic techniques including UV‐VIS, CP‐MAS ¹³C‐NMR, FT‐IR, and fluorescence spectroscopy. Clear chemical differences between the humic acid size fractions were observed. Smaller size fractions of the soil humic acid contained more chargeable functional groups and a larger percentage of aromatic carbon than the larger size fractions. Conversely, the percentage of aliphatic carbon increased with increasing apparent molecular weight. The chemical composition of the smallest humic acid fraction differed clearly from the fulvic acid fraction, despite similar apparent molecular size and carboxyl carbon content. Small humic acids contained much more aromatic carbon and less aliphatic carbon than the fulvic acid fraction. Apparently, humic size fractions differ in their chemical composition, which can have important implications for their environmental behaviour.     

Chemical–structural information from solid-state 13C NMR studies of a suite of humic materials from a lower montane forest soil,Colorado, USA

Chemically and physically fractionated samples extracted from the surface horizon of a soil developed under a mix of coniferous and deciduous vegetation in southwestern Colorado were studied. ¹³C NMR data on this soil's organic matter and its HF(aq)-washed residue, as well as the classic acid/base-separated humic fractions (humic acid, fulvic acid, humin), were examined for chemical–structural detail, e.g., the various structural functionalities present (especially lipids, carbohydrates, aromatics, polypeptides and carbonyl/carboxyls). Among the humic fractions, it was found that the lipid concentrations are in the order humic acid>fulvic acid= humin; for carbohydrates the order is fulvic acid>humin>humic acid; for aromatic carbons the order is humic acid>humin>fulvic acid; for polypeptides it is humic acid>fulvic acid>humin and for carbonyl/carboxyl species it is humin>humic acid>fulvic acid, but the differences are small. ¹³C spin–lattice relaxation times indicate that at least two types of “domains” exist in each, corresponding to “higher” and “lower” concentrations of paramagnetic centers, e.g., Fe³⁺.     

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.     

Incorporation of nitrogen from urea fertilizer into soil organic matter in rice paddy and cassava upland fields in Indonesia

Incorporation of newly-immobilized N into major soil organic matter fractions during a cropping period under paddy and upland cropping systems in the tropics was investigated in Jawa paddy fields with and without fish cultivation and a Sumatra cassava field in Indonesia. ¹⁵N-labelled urea (¹⁵N urea) was applied as basal fertilizer, and the soil samples were collected after harvest. The percentage of distribution of the residual N in soil from ¹⁵N urea into the humic acids, fulvic acid fraction, and humin were 13.1–13.9, 19.0–20.5, and 53.4–54.3%, respectively, for the Jawa paddy soils, and 14.9, 27.4, and 52.4%, respectively, for the Sumatra cassava soil. These values were comparable to the reported ones for other climatic zones. The percentage of distribution of ¹⁵N urea-derived N into humic acids was larger than that of total N into the same fraction in all the soils. The distribution into the fulvic acid fraction was also larger for ¹⁵N urea-derived N than for total N in the Jawa soils. Humic and non-humic substances in the fulvic acid fraction were separated using insoluble polyvinylpyrrolidone (PVP) into the adsorbed and non-adsorbed fractions, respectively. Less than 5% of the ¹⁵N urea-derived N in fulvic acid fraction was detected in the PVP-adsorbed fraction (generic fulvic acids). The proportion of non-hydrolyzable N remained after boiling with 6 M HCl in the ¹⁵N urea-derived N was 9.4–13.5%, 17.3–26.7%, and 8.4–16.6% for the humic acids, generic fulvic acids, and humin, respectively. The significantly low resistance to acid hydrolysis suggested that the ¹⁵N urea-derived N was less stable than the total N in soil regardless of the fractions of humus.     

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.     

Nitrogen use Rationalization and Boosting Wheat Productivity by Applying Packages ‎of Humic,Amino Acids,and Microorganisms

ABSTRACT

Integrated management of soil organic matter and nutritional status of crop plants is essential to sustain the production of organic farming systems. Thus, a 2–year field experiment was conducted to examine the effects of soil additions (192 kg N ha^–1, humic+192 kg N ha^–1, humic+144 kg N ha^–1 and humic+96 kg N ha^–1) and foliar applications (amino acids, Azotobacter+yeast, and amino acids plus Azotobacter+yeast) as various fertilizer resources on growth and yield of wheat. Results showed that humic+192 kg N ha^–1 × amino acids plus Azotobacter+yeast were the effective combination for producing the highest values of flag leaf area, total dry weight, tiller number m^–2, spike weight m^–2, and grain yield ha^–1. Under foliar application of amino acids plus Azotobacter+yeast, reducing N supply from recommended rate (192 kg N ha^–1) to 144 kg N ha^–1+ humic achieved higher values of all yield traits, with a saving of 25% of applied mineral nitrogen as well as enhancing nitrogen use efficiency.     

Humus of soils of the Altai Mountains

Data on the humus composition and specific features of soils in the Altai Mountains obtained in the long-term studies initiated by R.V. Kovalev are discussed. The average statistical values for the content of the main humus components and their ratios in different soil types were obtained by processing the data on 307 soil pits. The comparison of the soils belonging to the same type and occurring in the northwestern, central, and southeastern regions of the Altai Mountains in terms of the C ha/C fa ratio (one of the integral indices of the humus composition) showed that there were no significant differences between them. The overlapping intervals of the average values of this ratio testified to this fact. For instance, the average C ha/C fa ratios in the mountain tundra soils of the regions mentioned amounted to 0.70 ± 0.03; 0.72 ± 0.02; and 0.69 ± 0.03, respectively, and, in the mountain meadow soils, they amounted to 0.67 ± 0.03; 0.69 ± 0.04; and 0.67 ± 0.03, respectively. The mountain brown forest soils that are components of the soil cover only in the northwestern and central regions also differ insignificantly by this parameter (0.88 ± 0.05 and 0.89 ± 0.03, respectively). In the soils of the Altai Mountains, the dependence between the portion of humic acids and the mean annual air temperature (HA (%) = 29.54 + 1.06T(°C), r = 0.71) and the ratio of the portion of fulvic acids to the mean annual precipitation (FA (%) = 9.70 + 0.029W, r = 0.74) was shown to be similar to those in all the soils of mountainous southern Siberia. These facts enabled us to apply regression equations for a quantitative reconstruction of the paleoclimate components according to the humus composition. Original Russian Text ￠ M.I. Dergacheva, E.I. Kovaleva, N.N. Ryabova, 2007, published in Pochvovedenie, 2007, No. 12, pp. 1416–1421.     

Use of alkaline soil extracts for 13C n.m.r. characterization of humic substances

Previous solution ¹³C n.m.r. studies of soil organic matter have been confined to isolated humic or fulvic acid fractions. A study of 10 crude alkaline extracts from five New Zealand topsoils has now shown that such purification is unnecessary. Spectra were not noticeably broadened by co-extracted iron in concentrations up to 2 mg cm^?3. Optimized pulse parameters were similar to those reported for a solution of a purified humic acid. Relaxation results support a generalized model for humic material, in which aromatic structures, carbohydrates, amino acids and polymethylene chains are linked together to form flexible macromolecules. Relative contributions from each component varied from soil to soil.     

Impact of plant species and atmospheric CO2 concentration on rhizodeposition and soil microbial activity and community composition

Both plant species and CO₂ concentration can potentially affect rhizodeposition and consequently soil microbial activity and community composition. However, the effect differs based on plant developmental stage. We focused on the effect of three plant species (forbs, grasses, and N₂‐fixers) at an early stage of development on root C deposition and fate, soil organic matter (SOM) mineralization and soil microbial community composition at ambient (aCO₂) and elevated (eCO₂) CO₂ levels. Plants were grown from seed, under continuous ¹³C‐labelling atmospheres (400 and 800 µmol mol^?1 CO₂), in grassland soil for three weeks. At the end of the growth period, soil respiration, dissolved organic C (DOC) and phospholipid fatty acid (PLFA) profiles were quantified and isotopically partitioned into root‐ and soil‐derived components. Root‐derived DOC (0.53 ± 0.34 and 0.26 ± 0.29 µg mL soil solution^?1) and soil‐derived CO₂ (6.14 ± 0.55 and 5.04 ± 0.44 µg CO₂‐C h^?1) were on average two times and 22% higher at eCO₂ than at aCO₂, respectively. Plant species differed in exudate production at aCO₂ (0.11 ± 0.11, 0.10 ± 0.18, and 0.58 ± 0.58 µg mL soil solution^?1 for Plantago, Festuca, and Lotus, respectively) but not at eCO₂ (0.20 ± 0.28, 0.66 ± 0.32, and 0.75 ± 0.15 µg mL soil solution^?1 for Plantago, Festuca, and Lotus, respectively). However, no differences among plant species or CO₂ levels were apparent when DOC was expressed per gram of roots. Relative abundance of PLFAs did not differ between the two CO₂ levels. A higher abundance of actinobacteria and G‐positive bacteria occurred in unplanted (8.07 ± 0.48 and 24.36 ± 1.18 mol%) and Festuca‐affected (7.63 ± 0.31 and 23.62 ± 0.69 mol%) soil than in Plantago‐ (7.04 ± 0.36 and 23.41 ± 1.13 mol%) and Lotus‐affected (7.24 ± 0.17 and 23.13 ± 0.52 mol%) soil. In conclusion, the differences in root exudate production and soil respiration are mainly caused by differences in root biomass at an early stage of development. However, plant species evidently produce root exudates of varying quality affecting associated microbial community composition.