Quantitative aspects of solid-state C-NMR spectra of humic substances from soils of volcanic systems

Cross-Polarisation Magic Angle Spinning Carbon-13 Nuclear Magnetic Resonance spectroscopy (CPMAS ¹³C-NMR) represents one of the most powerful tools to investigate soil organic matter (SOM) mainly because of its inherent capacity to provide a semi-quantitative evaluation of carbon distribution. A critical parameter during acquisition of CPMAS ¹³C-NMR spectra is the contact time required to obtain the cross-polarisation between proton and carbon nuclei. The procedure to evaluate the best contact time for the acquisition of a quantitative CPMAS ¹³C-NMR spectrum is to perform Variable Contact Time (VCT) experiments. In this work the structural features of a number of purified humic substances from Italian and Costarican volcanic soils were investigated by CPMAS ¹³C-NMR spectroscopy after having performed preliminary VCT experiments. The VCT experiments showed that the average contact times vary according to the origin and chemical structure of the humic material. The optimal contact times (OCT) for nine humic samples were between 250 and 800 μs These values were different from the time of 1000 μs that is commonly applied as the best average contact time for humic materials. Moreover, by comparing the NMR data to those obtained by elemental analysis (C/H ratio), it appeared that the efficiency of the cross-polarisation between protons and carbons, and hence the contact time, is affected not only by the number of protons, but also by their distribution over the molecules. The evaluation of errors in quantitative estimation of the different carbons revealed that the use of OCT generally reduced by half the loss of signals occurring when the average contact time of 1000 μs is used in CPMAS ¹³C-NMR spectra of humic substances.     

土壤腐殖质氮结构的X-光电能谱研究

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.     

Fate of humic acids isolated from natural humic substances

Composition of humic acids (HA) is a function of plant-derived inputs, degradation processes regulated by microorganisms, organo-mineral interactions and age. Characterization of different origin humic substances is important for evaluation of their contribution to stabile and labile carbon pool in the environment. The relative abundance of chemical components in HA isolated from soils, compost, commercial lignohumates, alginite, acadiane and lignite was studied with aim to quantify content of important biomarkers such as amino acid, lipids and polyphenols. HA were considered as a heterogeneous complex and high concentration of peptides, polyphenols and lipids was determined in acadian-HA to compare with soil-HA. Compost-HA contained much more amino acids to compare with soil-HA samples. Alginite-HA and lignite-HA were similar in biomarkers content to soil-HA. Fourier transform infrared spectroscopy confirmed that chemical composition and functional groups content differs with the origin, humification degree and the age of studied samples. Soil-HA are typically composed of a variety of ?OH, COOH?, C–O, C–H₂, (aliphatic and aromatic) groups, quinines, lignin fragments, polysaccharide, monosaccharide and proteins fragments, which are linked together by ?O?, ?NH?, ?H=, >C=O, metal ions and –S? groups. ¹³C NMR spectroscopy showed that aromatic carbon content was the highest in lignite-HA and soil-HA.     

Characterization of fungal melanins and soil humic acids by chemical analysis and infrared spectroscopy

Summary Humic acids from two Brazilian topsoils under savanna grassland and five soil fungal melanins were characterized by elemental, functional group and infrared analysis. C, N, total acidity, COOH, and phenolic OH contents were within the ranges reported for several other fungal melanins and soil humic acids. Compared with the soil humic acids, the infrared spectra of the fungal melanins showed greater detail, indicative of higher aliphaticity. They were similar to the type III infrared spectra of humic acids, which are characteristically high in proteinaceous material and polysaccharides. The infrared spectra of the humic acids from the two Brazilian soils studied were classified as type I, which includes most soil humic acids. Notwithstanding the greater detail, in some areas the fungal melanin spectra were similar to those reported for other fungal melanins and humic acids of different origins. The probable contribution of the melanic fungi to the formation of soil humic polymers is discussed.     

Solubilization of iron by water‐extractable humic substances

The capability of water‐extractable humic substances (WEHS) to solubilize Fe from sparingly soluble Fe‐hydroxide was studied. Addition of WEHS (1.7 mmol organic C l^—1) to a dialysis tube containing labeled insoluble Fe‐hydroxide caused an increase in the amount of ⁵⁹Fe measured in the external solution. The humic fraction was also able to solubilize Fe from soil samples, with levels comparable to those obtained using a solution containing 100�μM DTPA. By measuring the amount of ⁵⁹Fe eluted from soil columns pre‐loaded with ⁵⁹Fe‐WEHS it was possible to evaluate the mobility of Fe complexed to the humic molecules. The recovery of ⁵⁹Fe varied from 2% to 25% in respect to the soil type used. The ability of Fe‐WEHS to serve as an Fe source for the phytosiderophore hydroxy‐mugineic acid (HMA) was also analyzed. The removal of ⁵⁹Fe from the Fe‐WEHS complex by HMA was demonstrated by adding the phytosiderophore to a dialysis tube containing the ⁵⁹Fe‐WEHS complex. The observations suggested a ligand exchange between the phytosiderophore and the humic fraction. The results indicate that WEHS is able to increase the amount of Fe present in the soil solution, possibly by forming mobile complexes with the micronutrient. These complexes could act as easily available Fe sources in Fe acquisition processes by both monocot and dicot plants, playing an important role particularly in soils with low available Fe.     

Characteristics of humic substances in peat of selected peatlands from north-eastern Poland

Abstract

Peat samples collected in six peatlands located in north-eastern Poland were analysed. Two of the investigated psedands were fens, two were transitional bogs and two of them were raised bogs. All peat deposits were investigated in the whole stratigraphic profile, and peat samples were chosen according to the differentiation of peat genus in deposit. pH in water and KCl, degree of decomposition, ash content, carbon content as well as the ratio of humic to falvic acid were evaluated. The highest degree of peat decomposition was found in wood peat (Alneti), and the in moss peat (Bryaleti). The strongest humification was observed in low peat of genus Limno-Phragmitioni (hypnum-moss peat) and Magnocaricioni (sedgeous peat).     

腐殖酸物质对磷在褐土中迁移的影响

采用室内土柱实验研究了腐殖酸和腐殖酸钠在与磷肥共施时对磷在褐土中迁移的影响。结果表明,与单施磷酸二氢钙相比,共施腐殖酸钠缩短了磷在土壤中的迁移距离,明显降低了施肥点附近微域中的水溶性磷和酸溶性磷含量,磷向土体中的迁移量也显著下降;相比之下,共施腐殖酸则明显增加了磷的迁移距离和迁移量,对肥际微域中不同形态磷的含量均有显著提高作用。因此,腐殖酸促进了磷在褐土中的迁移,适宜与磷肥配合施用,而腐殖酸钠不适合与磷肥共施。     

固态13C和15N核磁共振法研究15N标记土壤的腐殖质组分

Five humic fractions were obtained from a uniformly ^15N-labelled soil by extraction with 0.1 mol L^-1 Na4P2O7,0.1mol L^-1 NaOH ,and HF/HCl-0.1 mol L^-1 NaOH,consecutively,and analyzed by ^13C and ^15N CPMAS NMR (cross polarization and magic angle spinning nuclear magnetic resonace).Compared with those of native soils humic fractions studied as a whole contained more alkyls ,methoxyls and O-alkyls,being 27%-36%,17%-21%and 36%-40%,respectively,but fewer aromatics and carboxyls(bein 14%-20% and 13%-90%,respectively),Among those humic fractions ,the humic acid(HA)and fulvic acid(FA) extracted by 0.1 mol L^-1 Na4P2O7 contained slightly more carboxyls than corresponding humic fractions extracted by 0.1 mol L^-1 NaOH ,and the HA extacted by 0.1 mol L^-1 NaOH after treatment with HF/HCl contained the least aromatics and carboxyls.The distribution of nitrogen functional groups of soil humic fractions studied was quite similar to each other and also quite similar to that of humic fraction from native soils.More than 75% of total N in each fraction was in amide from,with 9%-13% present as aromatic and /or aliphatic amines and the remainder as heerocyclic N.     

Effects of coal derived humic substances on water retention and structural stability of Mediterranean soils

Abstract. The potential for using coal-derived humic substances to improve the available water holding capacity (AWC) and aggregate stability of typical Mediterranean soils was evaluated in the laboratory using an agricultural surface (0–20 cm) soil from each of three regions of Italy, (Sicily, Tuscany and Venetia) and five rates of humic acids (HA), 0,0.05,0.10,0.50 and 1.00 g/kg. There were significant ( P < 0.05) differences between the field capacity (FC), permanent wilting point (PWP), and available water capacity (AWC) values of the controls and those treated with 0.05 g/kg of the HA. Beyond this rate, differences in these properties were not significant. At the 1.00 g/kg HA rate, the relative improvements in AWC over the three controls were 30%, 10% and 26%. Low rates (0.05 to 0.10 g/kg) of HA were also needed to obtain a 40 to 120% improvement in aggregate stability of these soils relative to the controls. These results indicate that the addition of highly humified organic matter such as coal-derived humic substances can improve the structural and water retention properties of degraded arable soils. However, since there is not yet any direct evidence that these humic materials can ameliorate soils under field conditions, field studies will be needed to validate these results.     

Identification of organically bound iodine in soil humic substances by size exclusion chromatography / inductively coupled plasma mass spectrometry (SEC / ICP-MS)

Abstract

Sulfur transformation in riee rhizosphere was investigated. Soil enzyme arylsulfatase in rhizosphere and non-rhizosphere soil, whieh is responsible for mineralization of organic sulfur to sulfate sulfur, was studied. The Michaelis constants of arylsulfatase from Maahas c1ay and Pila c1ay loam were 3.04 × 10^-4 M and 3.97 × 10^-4 M, respectively. The arylsulfatase of rhizosphere soil showed higher activity than that of non-rhizosphere soil. Applieation of sulfate had no marked elTect on the enzyme aetivity either in rhizosphere or non-rhizosphere soil under the submerged condition. This indieates that arylsulfatase activity under the submerged condition is not inhibited by applieation of sulfate. The amount of HI-reducible sulfur in the rhizosphere and non-rhizosphere soi! inereased with time. However, rhizosphere soil had a higher amount of HI-reducible sulfur than did non-rhizosphere. Thc ditl'erence in arylsulfatase activity between the rhizosphcre and non-rhizosphere soil was not directly associated with thc number of sulfur-redueing and -oxidizing bacteria.     

Inhibitory action of dissolved humic substances on the growth of soil bacteria degrading DDT

Abstract

Dissolved humic acid (HA) and fulvic acid (FA) prepared from a Dando brown forest soil (Typic Dystrochrept) inhibited the growth of soil bacteria degrading DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) in the culture. The population of DDT-degrading Gram-variable rod Bll6 decreased by the application of both HA and FA, suggesting the presence of bactericidal effect. Such inhibitory effect was stronger for HA and resulted in a lower degrading activity of DDT in the culture of Bll6. No inhibitory effect was observed on the growth of DDT-degrading Bacillus sp. B75. The electron spin resonance spectra showed the presence of organic free radicals in both HA and FA. The relative concentration of the radicals was higher in HA. Storage of HA solution for 3 months at 4°C decreased the concentration of the radicals as well as the inhibitory action. The addition of catalase decreased the inhibitory effect of humic acid. It is suggested that a hydroxy radical, which is derived from free radicals of humic substances, is involved in the inhibition of bacterial growth and degradation of DDT.     

Mechanisms of plant growth stimulation by humic substances: The role of organo-iron complexes

Abstract

Stimulatory effects of humic substances (HS) on plant growth have been observed and widely documented. Studies have often shown positive effects on seed germination, root initiation and total plant biomass. The consistency of these observations has been uncertain, predominantly due to the lack of understanding of the plant growth promotion mechanism. Often these effects have been attributed to a direct effect of plant growth hormones; whereas in other instances the term “hormone-like activity” has been used to describe the plant growth stimulation (Chen and Aviad, Humic Substances in Soil and Crop Sciences: Selected Readings, American Society of Agronomy, Soil Science Society of America, 1990; Nardi et al., Humic Substances in Terrestrial Ecosystems, Elsevier Science B.V., 1996). Yet, investigators have been unable to prove that plant growth regulators are present in HS preparations, or the evidence provided remains unconvincing. An alternative hypothesis suggesting that growth enhancement of plants grown in nutrient solution (NS) containing HS is the result of improved micronutrient availability, Fe in particular, has been postulated and tested in the present study. Nutrient solutions containing N, P, K, Ca, Mg, S, B, Mo, Cu, Mn, Zn, and Fe at concentrations considered to be optimal for plant growth were tested for solubility of the Fe, Zn, and Mn, 7 days after preparation. In addition to control solutions at pH values of 5, 6, 7, and 7.5, 0 to 200 mg L^?1 of leonardite humic acid (HA) were added to the solutions and they were tested for Fe and Zn solubility. The HA greatly enhanced the maintenance in solution of Fe, in all the tested solutions, and Zn at pH 7.5. Mn mostly remained in solution in its inorganic forms. Plant growth experiments were carried out on both dicotyledonous plants (melons and soybean) and monocotytedonous Poaceae plants (ryegrass), due to the major difference in their Fe uptake mechanism. Plants grown in the absence of Fe exhibited severe Fe deficiency that could only partially be corrected with the addition of mineral Fe salts. The addition of HA or fulvic acid (FA) without addition of Fe, and Zn resulted in partial growth enhancement and correction of Fe deficiency, or none of the two, in the various experiments. This suggests that the growth enhancement effect observed in solutions containing Fe, Zn, and HS was related to the micronutrients rather than to phytohormones. However, the addition of Fe, Zn and either EDTA, HA or FA resulted in healthy, chlorophyll rich plants and enhanced growth, thereby providing evidence that improved Fe, and possibly Zn nutrition is a major mechanism of plant growth stimulation by HS. The use of the term hormone-like activity could be the result of the similarity of the physiological effects obtained in plants enjoying sufficient supply of Fe and Zn.     

Effect of cultivation on physical speciation of humic substances and plant nutrients in aggregate fractions of crusting soil from Zimbabwe

The effects of clearing and cultivation of Rhodic Kandiustalf from Mazowe (Zimbabwe) were studied by quantifying humic substances and plant nutrients (available and exchangeable cations) in aggregate fractions. It was found that cultivation was associated with a relative depletion of plant nutrients in coarse aggregate fractions (i.e. a virtual migration of soil fertility towards microaggregate soil compartments). This effect was connected with the organic matter distribution patterns in the physical fractions: cultivation has led to a relative concentration of the colloidal fractions (humic acids and fulvic acids) in the microaggregates, accompanied by a selective depletion of the latter organic fraction. It is suggested that the increased physicochemical importance of the microaggregate fractions after cultivation may be an index for the disruption of the original soil structure leading to crust formation and it may also be related to the effect of ploughing on the humification processes and the mobility of the colloidal organic fractions. Received: 1 August 1996     

Properties of humic substances from the baltic sea and lake ermistu mud

Scope and Goal  Humic substances (HSs) are the product of microbial degradation, chemical polymerisation and oxidation of organic matter. HSs can be divided into different fractions: humic acid (HA), hymatomelanic acid (HMA), fulvic acid (FA) and humin. HSs play a fundamental role in accumulation and exchange processes of chemical compounds (metals and organic pollutants) in the environment. HSs can form soluble complexes that migrate long distances or precipitate, carrying bound cations with them. The migration/precipitation abilities depend on the metal ion, the ion charge, the degree of ionisation of the organic molecule, the ionic strength of the media, and the location of the metal ion. Objective  Estonia is rich in the Baltic Sea and lake sediments. Historically, mud has been used in human treatment as a curative mud. High-molecular HSs are an important part of curative mud and they have a strong effect on its properties. The curative mud which is used in human therapy may not be polluted with different organic and inorganic contaminants. The aim of this work is to characterise and compare HSs isolated from the Baltic Sea mud (Haapsalu Bay) and from the sediments of Lake Ermistu (Estonia). Results and Conclusions  We determined the yield of basic extraction of different HSs components from mud in the course of separation. We found that acid pre-treatment of mud increased the amount of extracted HSs more in the sea mud than in the lake mud. These results show that HSs are bound to the inorganic/organic structure of mud and are released during prolonged treatment with an acid. We performed elemental analysis of the different fractions of HS extracts. HMA fractions had the highest carbon content and the lowest nitrogen content. HMA contains more polysaccharides than amino acid residues. These subunits may cause a better solubility of HMA in water as compared to HA. Acid pre-treatment of the natural sea and the lake mud diminished the content of carbon in most of the HS fractions. The content of nitrogen in the sea and in lake FA diminished by about two times. We determined the metal content in the mud and its HS fractions. We found that the concentration of heavy metals Pb and Cr is lower than <0.08 mg/kg. As expected, the total metal concentration is considerably lower in the lake than in the sea mud. Acid pre-treatment of mud shows that the sea HA forms more stable Fe and Mg complexes, while the sea HMA contains more stable Zn and Cu complexes. The lake HSs result in more stable Mg complexes with HA and HMA fractions. Sea FA binds Cu and Mg better, but lake FA is more effective in binding Zn. To compare the amount of metals extracted from HA, HMA, FA (with that, remains in HS fractions), the metal concentration in the alkaline full extract of the sea mud was determined. Alkaline treatment removed 0.8% Mg, 7.9% Cu, 5.2% Zn and 3.8% Fe together with HSs from the sea mud, the rest remained in humin and in the mineral part of the mud. The following work-up was additionally carried out for most of the metals. So, we found that there was only 1.7% of Fe, 2.1 % of Mg, 23.2% of Zn and 45.2% of Cu left in HA, HMA and the FA fraction (as a sum) from the total amount of those metals in HSs. This means that Cu is the metal most strongly bound to HSs. On the bases of HS separation data we found that the order of stability of the metal-humate complexes is Cu>Zn>Mg>Fe. The ultraviolet spectra of HA, HMA and FA revealed that HMA had the biggest molar absorption and calculated aromaticity. Recommendations and Outlook. The characteristics of HSs, isolated from the Baltic Sea and Lake Ermistu mud reveal the difference of HSs from the sea and the lake mud. Also, different properties of HS fractions are observed. Metals are concentrated variously in different HS fractions. In all cases the content of Pb and Cr was low, meaning that the mud preparations are nontoxic in respect to these metals. The order of stability of metal-humate complexes is Cu>Zn>Mg>Fe. The order of affinity of metals to HS fractions obtained have to be extended to other metals of environmental interest. The ability of HSs to bind metals may make them a candidate for natural, environmentally safe substances to concentrate hazardous metals and to remove them from natural water reservoirs.     

Effect of nitrate and humic substances of different molecular size on kinetic parameters of nitrate uptake in wheat seedlings

The kinetic parameters of nitrate uptake (I_max, Km and C_min) were evaluated in young seedlings of Triticum durum L., cv. Appulo, exposed to nitrate and/or to soil‐extracted humic acids (HAs) of different molecular weight. The uptake was enhanced after induction at low levels of nitrate (50 μM KNO₃), while it was inhibited after induction at higher concentrations (2000 μM). The kinetic parameters of uptake were selectively influenced by pre‐treatment with HAs: total (TE) and, at a greater extent, low (LMS, < 3500 Da) molecular size humic fraction increased either the nitrate uptake rate (I_max) and the efficiency of the whole transport system (low Km and C_min), while an opposite result was evidenced in high molecular size (HMS, > 3500 Da)‐treated plants. An additive effect was shown when nitrate and humic substances were provided simultaneously: the uptake rate was enhanced in TE‐ and LMS‐treated plants, but was strongly delayed in HMS‐treated plants. Removal of nitrate and/or humic fractions de‐induced the system and NO₃^— uptake rate decreased. Exposure to HAs was not able to induce nitrate reductase activity in root and leaf tissues. Inhibitors of protein synthesis p‐fluorophenylalanine and cycloheximide reversed the positive effect of LMS fraction on nitrate uptake. This would support the hypothesis of a promoting effect of HAs on the molecular expression of proteins of the nitrate transport system.     

腐殖酸对磷在红壤中有效性的影响

An investigation was conducted to study the effect of humic substance （HS） on the phosphorus （P） solubility in acidic soil. The soil （2.5 g）, HS （0, 0.5, and 2.5 g）, and P as monocalcium phosphate （0.31 and 1.25 g P kg^-1 soil） were mixed with 50 mL distilled water and two different sequences of adding HS and P were used. The results indicated that the P concentration in water and 0.01 mol L-1 CaCl2 solution increased with increasing amounts of humic substance. The concentrations of Fe and Al were also increased. However, Olsen P decreased with increasing amount of humic substance. Water-soluble P concentrations from P rates at 0.31 and 1.25 g P kg^-1 soil in the treatment with 0.5 g （2.5 g） humic substance addition were 360% and 70% （500% and 90%） higher, respectively, than those in the treatment with no humic substance addition. P extracted by 0.01 mol L^-1 CaCl2 in the treatments with 0.5 and 2.5 g humic substance addition was increased by 400% and 540%, respectively, compared with that in the treatment without humic substance at the rate of 0.31 g P kg^-1 soil, while the corresponding P concentrations were increased by 80% and 90% at the rate of 1.25 g P kg^-1 soil. The order of mixing humic substance and phosphate did not significantly affect desorbed P and labile P extracted with CaCl2.     

Formulation of humic substances coated fertilizer and its use to enhance K fertilizer use efficiency for tomato under greenhouse conditions

A greenhouse experiment was conducted at Land Resources Research Institute, NARC, Islamabad to examine the impact of humic substances (HSs) coating on potassium fertilizers use efficiency. Tomato variety “Rio Grande” was used. The treatments applied were T1= Control (N, P at 250 and100?mg kg^?1 respectively), T2?=?N, P?+?K at 200?mg kg^?1 as SOP, T3?=?N, P?+?K at 200?mg kg^?1 as NPK blend, T4?=?N, P?+?K at 200?mg kg^?1 HSs coated SOP) and T5?=?N, P?+?K at 200?mg kg^?1 HSs coated NPK blend. Results indicated a positive impact of sole and HSs coated products on agronomic traits, nutrient concentration, fruit quality traits, flower number, fruit umber, fruit weight, chlorophyll contents, fresh and dry biomass, tissue water contents, diameter and fruit mineral composition. The response of afore said traits to applied treatment varied.     

黑土、栗钙土和潮土胡敏酸分子结构的差异性分析

以北方典型的黑土、栗钙土和潮土提取的胡敏酸为研究对象,通过元素分析、傅里叶红外光谱仪及13C核磁共振仪对三者进行结构分析。不同胡敏酸的各种元素含量略有差别,各种官能团含量也不同。元素分析结果来看H/C原子比潮土最高,栗钙土次之,黑土最少;O/C的原子比三者没有明显的差别;C/N比呈现由高到低的次序分别为黑土、栗钙土、潮土。3种胡敏酸的红外光谱吸收峰相似,但吸收强度有差别,如3 352 cm-1和2 931 cm-1处潮土和栗钙土信号强度相当,黑土较低,而在2 580 cm-1处三者信号差异不明显,1 720,1 620,1 420,1 220 cm-1这4个吸收峰信号强弱均为黑土最强,栗钙土次之,潮土最弱。而从核磁共振图谱可以看出潮土结构中含有更多的多糖和蛋白质类结构单元;黑土有机质含量高,芳香化程度也较高。