Method for Estimating the Content of the Chloroform-Extractable Green Fraction (CEGF) in HCl-DMSO Extract of Soils

Chloroform-extractable green fraction (CEGF) was detected in the supernatant obtained by alkali precipitation from the HCI-dimethylsulfoxide (DMSO) extract of Pg-rich soil. In the alkaline solution, the color of CEGF was green and CEGF showed strong Pg-like absorption bands. Ultraviolet and visible (UV-VIS) spectral analysis and gel chromatography on Sephadex G-50 were performed to compare several properties between CEGF and Pg. CEGF, which was purified by gel chromatography on Bio-Beads SX-1, displayed strong absorption bands at 609, 562, 445 and 280 inn in the alkaline solution. These absorption bands were almost similar to those of Pg. Furthermore, the UV-VIS spectrum of CEGF in the organic solvents showed a similar characteristic pattern of 4,9-dihydroxyperylene-3,10-quinone (DHPQ), which was considered to be a chromophore of Pg. Based on the results of gel chromatography on Sephadex G-50, CEGF mainly consisted of two fractions, corresponding to the G₂ and G₃ fractions of Pg. These results indicated that the method for extracting CEGF from Pg-rich soil in the present study was easy and selective and that CEGF was one of the components of, or a closely related substance to Pg. A colorimetric method for the estimation of the CEGF content in soils was developed. The calibration curve of CEGF was linear over a wide range of contents from 2.75 to 220 mg L⁻¹. The CEGF content in twelve samples of various soils was examined. CEGF was detected in all the soil sampled (5 orders) including three samples (3 orders) where Pg was not detected, and the content ranged from 0.07 to 1.66 g kg⁻¹ (dry soil). Therefore, the method for estimating the CEGF content in soils developed in the present study was found to be suitable for various soil orders and it was assumed that CEGF occurred in various soil orders.     

Analysis of the green fraction of humic acids

The green fraction of humic acids (HAs), Pg, was fractionated by gel chromatography on Sephadex G-50. Repeated chromatography of the crude Pg obtained by the first chromatography of HA yielded a sharp peak (G₁) and two broad bands (G₂) of purified Pg. The recovery of Pg in G₁ and G₂ as percentages of the total Pg content of the HA were 16.2 and 14.5%, respectively. Distinct peaks were detected at 615, 570, 450 and 280 nm in the UV-visible spectra of both G₁ and G₂ in alkali solutions. Estimates of weight-average molecular weight were 9.7 * 10³ for G₁ and 1.23 * 10⁴ for G₂. In the ¹H-NMR spectra the percentage of hydroxyl groups of 4,9-dihydroxy-perylene-3, 10-quinone (DHPQ) nuclei, which are known to be the chromophore of Pg, was 4.1 and 4.5% of the total H, respectively. Based on these values and the H/C ratios, DHPQ-C was estimated as 28% of total C. ¹³C-NMR spectra obtained using inverse-gated decoupling yielded 40–41% aromatic C, which suggested the presence of aromatic rings other than DHPQ. All of the IR, ¹³C- and ¹H-NMR spectra indicated that the two purified Pg samples largely differed from each other in their contents of alkyl and polysaccharide components.     

Application of Near Infrared Reflectance Spectroscopy for the Assessment of Soil Quality in a Long‐Term Pasture

Abstract

The objective of the present study was to assess the ability of near infrared reflectance spectroscopy (NIRS) to analyze chemical soil properties and to evaluate the effects of different phosphorus (P) and potassium (K) fertilization rates on soil quality in different layers of a long‐term pasture. The NIRS calibrations were developed for humus, total Kjeldahl nitrogen (N_Kjeldahl), and several humic substances (HA1, “mobile” humic acids fraction; ΣHA, sum of humic acids; FA1, “mobile” fulvic acids; ΣFA, sum of fulvic acids, etc.) using soil samples of rather heterogeneous origin, collected during 1999–2003. Different spectral preprocessing and the modified partial least squares (MPLS) regression method were explored to enhance the relation between the spectra and measured soil properties. The equations were employed for the quality prediction of a sod gleyic light loam (Cambisol) in five PK fertilization treatments. The soil was sampled in 2000 and 2003 in three field replicates at depths of 0–10, 10–20, 20–30, and 30–50 cm, n=60 samples yr^?1. The best coefficients of correlation, R², between the reference and NIRS‐predicted data were as follows: for N_Kjeldahl, 0.965; humus, 0.938; HA1, 0.903; HA2, 0.905; HA3, 0.924; ΣHA, 0.904; and FA1, 0.911; and ΣFA, 0.885. Our findings suggest that it is feasible to use NIRS for the assessment of the effects of the inorganic PK fertilizer on the soil quality in different depths of a long‐term pasture.     

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.     

Depth distribution of humic substances in Andosols in relation to land management and soil erosion

Although Andosols are relatively resistant to water erosion, they can be severely affected by changes in land use, resulting in accelerated erosion and loss of soil organic matter (SOM). We hypothesized that if the contents of specific components of SOM and organo–metallic complexes (humic acids –HAs–, fulvic acids –FAs–, sodium pyrophosphate extractable carbon –C_p–, aluminium –Al_p–, and iron –Fe_p–) consistently tend towards certain ratios in A and B horizons, they could be used to identify soils denuded by erosion. To test this hypothesis, we investigated the vertical distribution of humus components and certain ratios, namely C‐HA/C‐FA, C‐FA/total organic C (TOC), C_p/TOC and (Fe_p + Al_p)/C‐FA, in representative profiles of andic soils located in natural ecosystems with different degrees of human disturbance. Furthermore, we analysed these parameters in the topsoil of a natural protected area and in adjacent soils under different land use scenarios (natural reserve vs. traditional exploitation). We found that the ratios of C‐HA/C‐FA and, to a lesser extent, of C‐FA/TOC and C_p/TOC changed with depth in the selected soil profiles, but the values were characteristic of each type of soil horizon. The values of these ratios in the topsoils of the disturbed areas were closer to a B horizon than an A horizon. This pattern may be superimposed on pre‐existing gradients, such as those related to the type of natural vegetation. The use of these indices emerges as a possible land use and erosion indicator.     

FTIR‐spectroscopic characterization of humic acids and humin fractions obtained by advanced NaOH,Na4P2O7, and Na2CO3 extraction procedures

Aim of our study was the development of the methodological basis for the characterization of humic fractions of a long‐term field experiment. Humic acids (HAs) were extracted from three layers of a nontilled soil using three different extractants (1 M NaOH, 0.1 M Na₄P₂O₇, 1 M Na₂CO₃), and the humin fraction was enriched. NaOH as extractant for FTIR analysis of humic substances yields higher resolved IR spectra, especially in the important regions of stretching vibrations including aromatic and aliphatic groups and in the fingerprint area including amides, aliphats, and aromats than the other extractants. The NaOH extraction has lower extraction yields as compared to Na₄P₂O₇ and Na₂CO₃ and represents a different part of the soil organic matter (SOM). This is reflected by lower C : N ratios and higher E₄ : E₆ and fulvic acid–to–humic acid ratios as compared to the other extractants. The FTIR band areas of HA fraction obtained by NaOH showed an increase of the aromatic and carbonyl groups and a decrease of amide groups with increasing soil depth. Aliphatic groups showed contradicting results: The bands of the stretching vibrations increased, and the band of the bending vibrations decreased. We assume that band interactions in the bending vibrations were responsible for that phenomenon under the assumption of an increase of aliphatic groups with increasing soil depth. The IR bands of the enriched humin fraction showed a decreasing trend in case of both aliphatic bands deriving from stretching vibrations and an increase of aromatic characteristics with depth. Our study led to the conclusion that HA fractions obtained by 1 M NaOH represent a small and dynamic fraction indicated by the measured yields in combination with values of N_t, C : N, E₄ : E₆ ratios, and ratios of fulvic acids (FA) to HA. The humin fraction has a high contribution to the total organic C and represents a more stabilized fraction of SOM which still shows changes in its aromatic and aliphatic characteristics with soil depth.     

Enzymatic transformation of humic substances by NDO

Enzymatic transformation of humic acids (HA), fulvic acids (FA) and indole was examined using naphthalene 1,2-dioxygenase (NDO). NDO was used as a model for dioxygenase enzymes found in various microbial species. Indole was used as a model substrate for NDO-catalyzed reactions resulting in condensation products. Although NDO is not classified as a soil enzyme, all HA and FA tested were susceptible to NDO-induced transformation. The extent of NDO-specific NADH oxidation in solutions containing HA and FA paralleled the percent aromaticity of the HA and FA. Furthermore, the UV–Vis absorptive properties of NDO-treated HA and FA were altered in a manner suggesting condensation reactions similar to the formation of indigo from indole. Condensation reactions were enhanced in NDO-treated mixtures containing indole and an FA. NDO retained activity for 2 weeks under ambient conditions, and retained some enzymatic activity for 9 days based on detection of specific metabolites by HPLC, suggesting prolonged extracellular activity. Humic substances have not previously been known to be substrates for dioxygenases; even more significant was that dioxygenase enzymes can facilitate condensation reactions between indole-like functional groups well-known to be present in HA and FA. These results illustrate how dioxygenases can be potential humic-modifying enzymes when released into the environment upon microbial death and concurrent cell lysis which could alter the bioavailability of organic contaminants associated with dissolved organic matter through specific modulation of enzyme activity involving substrate competition.     

Soil organic matter composition as a factor affecting the accumulation of polycyclic aromatic hydrocarbons

Journal of Soils and Sediments - The objective of this study was to evaluate the mutual relations between the soil organic matter (SOM) fractions: fulvic acids (FA), humic acids (HA), humins (HN),...     

Abstracts of Nippon Nojohiryogaku Zasshi

Abstract

P type humic acid which has absorption bands near 615, 570, and 450 run due to the presence of dihydroxyperylenequinone (DHPQ) was investigated by gel-chromatography. Humic acid samples were fractionated using Sephadex G-50 as gel material and 0.1 n NaOH, with or without 1.5 m urea, as eluent. The absorption spectra of the effluent fractions were measured, and elution curves were drawn on the basis of the optical density at 600 nm.

In general, the intensity of the absorption bands due to DHPQ increased with increasing fraction number, accompanied by a change in effluent color from brown, greenish brown to green. Based on the elution curves, the effluent fractions of a humic acid containing a large amount of the green fraction were divided into six groups: B₁, B₂, BG, G₁, G₂, and G₃. These fraction groups, especially the G groups and cercosporin, a dihydroxyperylenequinone pig ment, were compared by absorption spectroscopy, ultrafiltration, ultimate analysis, TLC, and gel-chromatography.

The green fraction was composed of a non-humic, relatively lower-molecular weight polymer containing a considerable amount of DHPQ nucleus, while the brown humic fraction still maintained the characteristic light absorption due to DHPQ. It may be tentatively con cluded that the green fraction is not only a co-extracted impurity, but is also firmly associated with dark-colored humic substances.     

Influence of natural and synthetic humic substances on the activity of urease

The activity of a purified urease, obtained from Bacillus pasteurii, was inhibited by humic and fulvic acids obtained from an agricultural soil. Enzyme kinetic studies showed that the humic substances affected the affinity of the enzyme for its substrate (K_m) and the maximum velocity of the reaction (V_max). The V_max was inhibited to the same extent by both humic (HA) and fulvic (FA) acids, the precise effect depending on the pH and concentration of humic substance. At pH 4.0, HA concentrations of 25 pg cm^?3 and 10 μg cm^?3 inhibited the V_max by 38.5% and 20% respectively. HA and FA had similar effects on the K_m but in this case the lowering of the affinity of the enzyme for its substrate was not concentration dependent in the range 0–25 μg cm^?3 of humic substance. Typically, the affinity was decreased from a K_M of 50 mM in the control to 67 mM in the presence of HA and FA. The effects were not due primarily to the ash or N contents of the humic substances because de-ashed humic acid and synthetic model humic (made from catechol, guaiacol, pyrogallol, resorcinol and protocatechuic acid) and fulvic acid (made from polymaleic acid), containing virtually no ash or N, were equally as effective. The effect was not related to the phenolic monomers which, before polymerization, had no effect on urease activity.     

Carbon and nitrogen in operationally defined soil organic matter pools

Humic substances [humic acid (HA), fulvic acid (FA), and insoluble humin], particulate organic matter (POM), and glomalin comprise the majority (ca 75%) of operationally defined extractable soil organic matter (SOM). The purpose of this work was to compare amounts of carbon (C) and nitrogen (N) in HA, FA, POM, and glomalin pools in six undisturbed soils. POM, glomalin, HA, and FA in POM, and glomalin, HA, and FA in POM-free soil were extracted in the following sequence: (1) POM fraction separation from the soil, (2) glomalin extraction from the POM fraction and POM-free soil, and (3) co-extraction of HA and FA from the POM fraction and POM-free soil. Only trace amounts of HA and FA were present in the POM fraction, while POM-associated glomalin (POM-glomalin) and POM alone contributed 2 and 12%, respectively, of the total C in the soil. Mean combined weights for chemically extracted pools from POM and from POM-free soil were 9.92 g glomalin, 1.12 g HA, and 0.88 g FA kg⁻¹ soil. Total protein and C, N, and H concentrations showed that glomalin and HA were, for the most part, separate pools, although protein was detected in HA extracts. Even though percentage carbon was higher in HA than in glomalin, glomalin was a larger (almost nine times) operationally defined pool of soil organic C. Glomalin was also the largest pool of soil N of all the pools isolated, but all pools combined only contained 31% of the total N in the soil.     

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.     

Macromolecule‐P associations and inositol phosphates in some chilean volcanic soils of temperate regions

Abstract

Total organic P, humic and fulvic acid‐P associations and inositol phosphates in nine volcanic soils of southern Chile were determined. The concentration of organic P (Po) ranged from 654 to 1942 ppm accounting for 49% to 64% of total soil P. Phosphorus associated to humic (HA‐P) and fulvic acids (FA‐P) accounted for 51–68% and 32–49% of Po, respectively. Inositol penta‐ and hexaphosphates represented 42% to 67% of Po suggesting that significant amounts are associated with both humic and fulvic acids. Po content was significantly correlated to organic C, total soil P and HA‐P. HA‐P and FA‐P fractions obtained from the most representative soil were examined by dyalisis and gel filtration. While approximately 96% of HA‐P presented a molecular weight higher than 100,000 daltons, 53% of FA‐P had a molecular weight under 12,000 daltons. It is suggested that these more labile organic P forms would be more easily mineralized, thus increasing the available P pool.     

三江平原典型湿地土壤腐殖质的剖面分布及其组成特征

以三江平原腹地挠力河、别拉洪河、浓江河流域自然沼泽湿地为研究对象,研究了典型湿地土壤腐殖质的剖面分布及其组成特征,并探讨了土壤腐殖质与植被类型及土壤全氮含量的关系。三江平原典型湿地土壤草根层和泥炭层中胡敏酸(HA)和富里酸(FA)的累计含量在整个剖面总量中的比例均大于70%,各组分相对含量分别高于35%总有机碳和23%总有机碳,其剖面分布与土壤有机碳(SOC)的变化趋势一致,由表层向下层逐渐减少。除小叶章湿地土壤草根层HA/FA值为0.97外,三江平原典型湿地土壤HA/FA均大于1,其剖面均值为1.4~2.5,表明三江平原湿地为胡敏酸型土壤.腐殖质各组份含量及其HA/FA值因植被类型而异,而与土壤全氮含量呈显著线性相关(p<0.05)。     

Modifications of degradation-resistant soil organic matter by soil saprobic microfungi

Modifications of humic (HA) and fulvic (FA) acids in their solutions and in sterile soil by microfungal species and two well-known HA degraders were studied by measurement of total oxidizable carbon (OC), absorbances, enzyme activities and CO₂ release. The effect of glucose on FA and HA, and also minerals on FA utilization was also observed. Microfungi affected HA more than FA. Common microfungal species decolorized HA and decreased their molecular size (evaluated in terms of A₄/A₆ ratio). Some of them decreased aromaticity of HA and FA as the only carbon sources. They did not affect OC, although released CO₂ from FA. Under higher availability of mineral nutrients, the FA aromaticity increased and FA decolorization decreased. The molecular size of HA decreased in the presence of glucose. In the FA medium complemented by minerals, the known basidiomycete HA degrader, Trametes versicolor, decreased the amount of aromatic compounds in contrast to microfungal species Alternaria alternata, Clonostachys rosea, Exophiala cf. salmonis, Fusarium coeruleum, F. redolens, Penicillium canescens, Phoma sp. and another basidiomycete Phanerochaete chrysosporium. No microfungal species exhibited lignin peroxidase activity. On the other hand, activities of manganese peroxidase (MnP) were recorded for all species incubated in FA. Carbon dioxide produced from soil inoculated by microfungi negatively correlated with the decolorization, aromaticity and OC of/in FA reisolated from the soil. The results support the hypothesis that soil microfungi can attack both HA and FA and can represent an important factor in their transformations in arable soils. The enzyme involved in FA modifications is probably fungal MnP. We enriched a group of known HA and FA degraders and showed some abilities of a few frequent soil microfungal species. This can be one of the first but important step towards learning the functioning of carbon release from the big reservoir represented by humic substances in arable soils.     

施用有机物料对土壤镉形态的影响

采用室内培养试验,研究作物新鲜秸秆和腐熟猪粪对模拟镉（Cd）污染的土壤中Cd形态转化的动态影响。结果表明,各处理土壤交换态Cd含量随培养时间均逐渐降低。碳酸盐结合态和铁锰氧化物结合态Cd含量先增加后降低, 而有机质结合态和残渣态Cd含量则逐渐增加。添加秸秆可增加土壤交换态Cd含量,但随时间延长,增幅逐渐降低, 猪粪则可降低土壤交换态Cd含量。添加有机物后土壤交换态Cd含量的变化主要是由有机质结合态或残渣态Cd含量的变化而引起。秸秆和猪粪对土壤Cd形态的转化与土壤胡敏酸（HA）和富里酸（FA）的变化有关。秸秆对能活化土壤Cd的FA增加幅度大于对能钝化土壤Cd的HA增加幅度,降低HA/FA比,但降幅随时间逐渐减少; 猪粪在整个培养阶段对HA增加幅度均大于FA的增加幅度,增加HA/FA比。秸秆和猪粪均可降低潮土pH而提高红壤pH,但只有猪粪可通过提高红壤pH降低Cd向交换态转化。添加秸秆和猪粪后,Cd由低活性态向交换态转化与HA/FA呈显著负相关。     

Characterization of humic substances derived from swine manure-based compost and correlation of their characteristics with reactivities with heavy metals

Various composts contain a significant amount of humic substances including humic acid (HA) and fulvic acids (FAs). The FA fraction in soils is considered to be sensitive to agronomic and environmental factors. In this study, three fractions of humic substances, HA (MW > 1000 Da), FA (MW > 1000 Da), and FA (MW < 1000 Da) were extracted from swine manure-based compost and characterized, and then, their reactivities were correlated with heavy metals. Compositions of the three fractions of humic substances were characterized by elemental and total acidity analyses and electron spin resonance (ESR), Fourier transform infrared (FTIR), and 13C nuclear magnetic resonance with cross-polarization and magic-angle spinning spectroscopic techniques. Elemental analyses indicated that HA has higher contents of C, H, N, and S than those of FAs. However, FA (MW > 1000) and especially FA (MW < 1000) have higher contents of O than that of HA (MW > 1000). The g values of the ESR spectra of the three fractions showed that the organic free radical characteristics and the widths of the spectra and free radical concentrations of the three fractions are significantly different. The FTIR spectra indicated that HA (MW > 1000) is abundant in C=C bonds while FA (MW > 1000), especially FA (MW < 1000), are abundant in C=O bonds. In addition, 13C NMR spectra indicate that carboxyl contents of FA (MW > 1000), especially FA (MW < 1000), are higher than that of HA (MW > 1000). The sequence of the reactivity in terms of acidic functional groups was FA (MW < 1000) > FA (MW > 1000) > HA (MW > 1000). Elemental and functional group compositions of the three fractions significantly correlated with reported reactivities with heavy metals. The application of swine manure-based compost containing HA and FAs fractions to soil and associated environments may thus significantly affect the concerned reactions with organic and inorganic compounds including pollutants.     

Relationship between molecular characteristics of soil humic fractions and glycolytic pathway and krebs cycle in maize seedlings

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.     

Characterization of water soluble organic matter in soils by size exclusion chromatography and fractionation with polyvinylpyrrolidone

Water extracts were obtained from four types of soils (Brown Lowland soil, Yellow soil with manure application for 6 years, non-allophanic Andosol, and allophanic Andosol), and the organic matter in the water extracts was fractionated according to the solubility in acid and adsorption onto polyvinylpyrrolidone (PVP). For the water extracts and their fractions, the amounts of organic C, total N, and anthrone-reactive C (ARC) were analyzed, and high performance size exclusion chromatography (HPSEC) was carried out. The PVP-non-adsorbed fulvic acid (FA) fraction accounted for the largest proportion of the total water-soluble organic C, ranging from 52% for the Yellow soil to 96% for the allophanic Amdosol, followed by the PVP-adsorbed FA and humic acid (HA) fractions. The water extract of the allophanic Andosol was characterized by the lack of HA fraction and a small proportion of PVP-adsorbed FA fraction. For all the water extract samples, more than 90% of the ARC was recovered in the PVP-non-adsorbed FA fraction. The proportion of ARC in the total organic C in the fraction was also highest in the PVP-non-adsorbed FA fraction. The molecular weight (MW) of the humic substances (HS) at peak maximum was estimated at 1,300 Da for the water extracts and their fractions from the Brown Lowland soil and non-allophanic Andosol samples by HPSEC using polyethylene glycols as MW standards. Manure application increased the MW of HS in the HA and PVP-adsorbed FA fractions. On the other hand, only a small amount of HS was found in the water extract of the allophanic Andosol by HPSEC.     

The significance of solid‐state 13C NMR spectroscopy of whole soil in the characterization of humic matter

Abstract

Differences in characteristics of humic matter were investigated by solid‐state CP/MAS ¹³C NMR spectroscopy of whole (nontreated) materials and their extracted humic fractions. Samples used in the analysis were lignite, a commercial humate AG, and the Bh horizons of a Mascotte and a Lawnwood soil. Humic fractions were extracted by the 0.1 M NaOH or Na₄P₂O₇ (pH 9.8) method. The humic (HA) and fulvic acid (FA) obtained were weighed and analysed for total acidity, carboxyl and phenolic‐OH group contents. Whole lignite, humate AG and soil samples, and the HA and FA fractions were analyzed by solid state CP MAS ¹³C NMR and infrared spectroscopy. Carbon, H, and N contents were determined by chemical analysis. NMR spectra of the combined HA+FA extracts resembled the spectra of the whole materials. No additional signals were detected, indicating that alien compounds were not produced during the extraction. The best spectra were obtained with HA samples produced by the NaOH method. These spectra closely resembled those of the untreated materials. Spectral and chemical differences noticed between the HA (or FA) fractions were attributed more to differences in origin than to the extraction procedure. Aliphatic, aromatic and carboxyl groups were the major components of HA from lignite and humate AG. In contrast, HA from the two Haplaquods were characterized by four major components: the aliphatic, polysaccharide, aromatic, and carboxyl groups. Regardless of origin, all the HA fractions contained similar functional groups, as indicated by their close similarities in infrared spectra.