Enhanced washing of polycyclic aromatic hydrocarbons from contaminated soils by the empowered surfactant properties of de novo O-alkylated humic matter.

Aqueous solutions of humic acid (HA) derivatized by a catalyzed O-alkylation reaction with methyl, pentyl, and benzyl groups at 40, 60, and 80% of total HA acidity were used to wash off polycyclic aromatic hydrocarbons (PAHs) from two contaminated soils. The enhanced surfactant properties enabled the alkylated HA to remove phenanthrene, anthracene, fluoranthene, and pyrene from both soils more extensively than the original unmodified HA, the 60% benzylation generally showing the greatest soil washing efficiency. For both soils, all alkylated HA revealed greater PAH removals than Triton X-100 nonionic surfactant, while the benzylated and methylated HA nearly and fully matched pollutants release by the anionic SDS in the coarse- and fine-textured soils, respectively. A consecutive second washing with 60% benzylated HA removed additional PAHs, in respect to the first washing, from the coarser-textured soil, except for fluoranthene, while removal from the finer-textured soil incremented even more for all PAHs. These findings indicate that the enhanced hydrophobicity obtained by a simple and unexpensive chemical derivatization of a natural humic surfactant can be usefully exploited in the washing of polluted soils, without being toxic to the soil biota and by potentially promoting the subsequent bio-attenuation of organic pollutants.

Varying the hydrophobicity of humic matter by a phase-transfer-catalyzed O-alkylation reaction.

An O-alkylation reaction catalyzed by tetrabutylammonium hydroxide (TBAH) as a phase-transfer agent was applied to a humic acid (HA) to modify its hydrophobic properties. The carboxyl and hydroxyl functional groups of HA acted as nucleophiles in substitution reactions (Sn2) with methyl iodide, pentyl bromide and benzyl bromide added in amounts equimolar to 20, 60 and 80% of HA total nucleophilic sites. The occurrence of O-alkylation was shown by DRIFT spectrometry, NMR spectroscopy, High Performance Size Exclusion Chromatography (HPSEC) and elemental analysis of reaction products. DRIFT spectra showed changes in C-H stretching and bending regions following the insertion of methyl and pentyl groups, while the incorporation of benzyl groups revealed the characteristics aromatic C-H stretching bands. Both liquid- and solid-state NMR spectra revealed characteristic signals for alkyl/aryl esters and ethers. HPSEC chromatograms of alkylated materials invariably displayed an increase in hydrodynamic volume in respect to the original HA, thereby suggesting that the enhanced hydrophobicity conveyed further associations among humic molecules. Analytical, HPSEC and spectroscopic results suggest that benzylation was the most effective reaction at all percentages of HA total nucleophilicity, followed, in the order, by pentylation and methylation, The benzylation reaction was used to improve reaction and work-up conditions and show that HA could be efficiently alkylated also with substantial reduction of TBAH amount, with no THF addition, increase of reaction time and of washing cycles to remove catalyst impurities. These findings indicate that the hydrophobicity of humic substances can be modulated through a mild O-alkylation reaction under a phase-transfer catalysis according to the extent of exposed HA nucleophilic sites. Such a structural modification of humic matter may have multiple chemical, environmental and biological applications.

More microbial manipulation and plant defense than soil fertility for biochar in food production: A field experiment of replanted ginseng with different biochars.

The role of biochar-microbe interaction in plant rhizosphere mediating soil-borne disease suppression has been poorly understood for plant health in field conditions. Chinese ginseng (Panax ginseng C. A. Meyer) is widely cultivated in Alfisols across Northeast China, being often stressed severely by pathogenic diseases. In this study, the topsoil of a continuously cropped ginseng farm was amended at 20 t ha-1, respectively, with manure biochar (PB), wood biochar (WB), and maize residue biochar (MB) in comparison to conventional manure compost (MC). Post-amendment changes in edaphic properties of bulk topsoil and the rhizosphere, in root growth and quality, and disease incidence were examined with field observations and physicochemical, molecular, and biochemical assays. In the 3 years following the amendment, the increases over MC in root biomass were parallel to the overall fertility improvement, being greater with MB and WB than with PB. Differently, the survival rate of ginseng plants increased insignificantly with PB but significantly with WB (14%) and MB (21%), while ginseng root quality was unchanged with WB but improved with PB (32%) and MB (56%). For the rhizosphere at harvest following 3 years of growing, the total content of phenolic acids from root exudate decreased by 56, 35, and 45% with PB, WB, and MB, respectively, over MC. For the rhizosphere microbiome, total fungal and bacterial abundance both was unchanged under WB but significantly increased under MB (by 200 and 38%), respectively, over MC. At the phyla level, abundances of arbuscular mycorrhizal and Bryobacter as potentially beneficial microbes were elevated while those of Fusarium and Ilyonectria as potentially pathogenic microbes were reduced, with WB and MB over MC. Moreover, rhizosphere fungal network complexity was enhanced insignificantly under PB but significantly under WB moderately and MB greatly, over MC. Overall, maize biochar exerted a great impact rather on rhizosphere microbial community composition and networking of functional groups, particularly fungi, and thus plant defense than on soil fertility and root growth.

Improved ginseng production under continuous cropping through soil health reinforcement and rhizosphere microbial manipulation with biochar: a field study of Panax ginseng from Northeast China.

The production of ginseng, an important Chinese medicine crop, has been increasingly challenged by soil degradation and pathogenic disease under continuous cropping in Northeast China. In a field experiment, an Alfisol garden continuously cropped with Chinese ginseng (Panax ginseng C. A. Meyer) was treated with soil amendment at 20 t ha-1 with maize (MB) and wood (WB) biochar, respectively, compared to conventional manure compost (MC). Two years after the amendment, the rooted topsoil and ginseng plants were sampled. The changes in soil fertility and health, particularly in the soil microbial community and root disease incidence, and in ginseng growth and quality were portrayed using soil physico-chemical assays, biochemical assays of extracellular enzyme activities and gene sequencing assays as well as ginsenoside assays. Topsoil fertility was improved by 23% and 39%, ginseng root biomass increased by 25% and 27%, and root quality improved by 6% and 18% with WB and MB, respectively, compared to MC. In the ginseng rhizosphere, fungal abundance increased by 96% and 384%, with a significant and insignificant increase in bacterial abundance, respectively, under WB and MB. Specifically, the abundance of Fusarium spp. was significantly reduced by 19-35%, while that of Burkholderia spp. increased by folds under biochar amendments over MC. Relevantly, there was a significant decrease in the abundance proportion of pathotrophic fungi but a great increase in that of arbuscular mycorrhizal fungi, along with an enhanced microbial community network complexity, especially fungal community complexity, under biochar amendments. Thus, biochar, particularly from maize residue, could promote ginseng quality production while enhancing soil health and ecological services, including carbon sequestration, in continuously cropped fields.

Macroaggregates Serve as Micro-Hotspots Enriched With Functional and Networked Microbial Communities and Enhanced Under Organic/Inorganic Fertilization in a Paddy Topsoil From Southeastern China.

Microbial communities of soil aggregate-size fractions were explored with molecular and networking assays for topsoil samples from a clayey rice paddy under long-term fertilization treatments. The treatments included no fertilizer (NF) as control, chemical fertilizer only (CF), chemical fertilizer with swine manure (CFM), and chemical fertilizer with rice straw return (CFS). Following a wet-sieving protocol, water-stable aggregates were separated into size fractions of large macroaggregates (L-MacA, >2,000 µm), macroaggregates (MacA, 2,000-250 µm), microaggregates (MicA, 250-53 µm), fine microaggregates (F-MicA, 53-2 µm), and fine clay (F-Clay, <2 µm). Mass proportion was 32.3-38.2% for F-MicA, 23.0-31.5% for MacA, 19.0-23.1% for MicA, 9.1-12.0% for L-MacA, and 4.9-7.5% for F-Clay, respectively. The proportion of MacA was increased, but F-Clay was reduced by fertilization, whereas the mean weight diameter was increased by 8.0-16.2% from 534.8 µm under NF to 621.5 µm under CFM. Fertilization affected bacterial 16S rRNA and fungal 18S rRNA gene abundance in F-MicA and F-Clay but not in aggregates in size larger than 53 µm. However, bacterial and fungal community α-diversities and community structures were quite more divergent among the fertilization treatments in all size fractions. Organic carbon and gene abundance of bacteria and fungi were enriched in both L-MacA and MacA but depleted in F-Clay, whereas microbial Shannon diversity was rarely changed by fraction size under the four treatments. L-MacA and MacA contained more bacteria of r-strategists and copiotrophs, whereas F-MicA and F-Clay were demonstrated with a higher abundance of K-strategists and oligotrophs. Guilds of parasitic and litter saprotrophic fungi were enriched in F-MicA but depleted in L-MacA. Furthermore, most of bacterial and fungal operational taxonomic units were strongly interacted in L-MacA and MacA rather than in MicA and F-Clay. Thus, MacA acted as micro-hotspots enriched with functional and networked microbial communities, which were enhanced with organic/inorganic fertilization in the rice paddy.

Phosphate and Ammonium Removal from Wastewaters Using Natural-Based Innovative Bentonites Impacting on Resource Recovery and Circular Economy.

The research objective of the study is the estimation of a novel low-cost composite material f-MB (Fe-modified bentonite) as a P and N adsorbent from wastewaters. Τhe present study aimed at examining the phosphate and ammonium removal efficiency from different types of wastewater using f-MB, by conducting bench-scale batch experiments to investigate its equilibrium characteristics and kinetics. The SEM analysis revealed that the platelets of bentonite in f-MB do not form normal bentonite sheets, but they have been restructured in a more compact formation with a great porosity. Regarding the sorption efficiencies (Qm), the maximum phosphate sorption efficiencies (Qm) calculated using the Langmuir model were 24.54, 25.09, 26.13, 24.28, and 23.21 mg/g, respectively, for a pH range of 5 to 9. In addition, the maximum NH4+-N adsorption capacities (Qm) calculated from the Langmuir model were 131.8, 145.7, 168.5, 156.7, and 159.6 mg/g, respectively, for a pH range from 5 to 9. Another important finding of this study is that f-MB can recover P from treated wastewater impacting on resource recovery and circular economy (CE). The modified clay f-MB performed the phosphate and ammonium recovery rates of 80% and 78.5%, respectively. Finally, f-MB can slowly release the largest proportion of phosphate and ammonium ions for a long time, thus extending the application of the f-MB material as a slow-release fertilizer and soil improver.

Quantitative assessment of the effects of biochar amendment on photosynthetic carbon assimilation and dynamics in a rice-soil system.

Biochar amendment has been proposed as a promising means to increase carbon (C) sequestration and simultaneously benefit plant productivity. However, quantifying the assimilation and dynamics of photosynthetic C in plant-soil systems under biochar addition remains elusive. This study established two experimental factors involving biochar addition and nitrogen (N) fertilization to quantitatively assess the effect of biochar on photosynthetic C fate in a rice plant-soil system. The rice plants and soil samples were collected and analyzed after 6-h pulse labeling with 13 CO2 at the tillering, jointing, heading and ripening stages. Biochar did not affect the proportions of photoassimilated carbon-13 (13 C) allocations in plant-soil systems. Nevertheless, biochar enhanced the 13 C contents in the shoot, root, and soil pools, especially when combined with N fertilization, and biochar increased the cumulative assimilated 13 C contents in the shoot, root, and soil pools by 23%, 14% and 20%, respectively, throughout the whole growth stage. Moreover, biochar addition significantly enhanced the N use efficiency (NUE) by c. 23% at the heading and ripening stages. In summary, biochar increases the content of photoassimilated C in plant-soil systems by improving plant productivity via enhancing NUE, thus resulting in a higher soil C sequestration potential.

Could biochar amendment be a tool to improve soil availability and plant uptake of phosphorus? A meta-analysis of published experiments.

As one of the most important nutrients for plant growth, phosphorus was often poorly available in soil. While biochar addition induced improvement of soil structure, nutrient and water retention as well as microbial activity had been well known, and the effect of biochar soil amendment (BSA) on soil phosphorus availability and plant P uptake had been not yet quantitatively assessed. In a review study, data were retrieved from 354 peer-reviewed research articles on soil available P content and P uptake under BSA published by February 2019. Then a database was established of 516 data pairs from 86 studies with and without BSA in agricultural soils. Subsequently, the effect size of biochar application was quantified relative to no application and assessed in terms of biochar conditions, soil conditions, as well as experiment conditions. In grand mean, there was a significant and great effect of BSA on soil available P and plant P uptake by 65% and 55%, respectively. The effects were generally significant under manure biochar, biochar pyrolyzed under 300 °C, soil pH <5 and fine-textured soil, and soils that are very low in available P. Being significantly correlated to soil P availability (R2=0.29), plant P uptake was mostly enhanced with vegetable crops of high biomass yield. Overall, biochar amendment at a dosage up to 10 t ha-1 could be a tool to enhance soil availability and plant uptake of phosphorus, particularly in acid, heavy textured P-poor soils.

Amendment of straw biochar increased molecular diversity and enhanced preservation of plant derived organic matter in extracted fractions of a rice paddy.

While biochar enhanced carbon sequestration and stability of soil organic matter (SOM), changes in organic molecular composition in biochar-amended soils had been poorly addressed. In this study, molecular composition changes of a paddy topsoil 2 years following amendments at 10 t ha-1 OC equivalent with untreated (CS), manured (CM) and charred (CB) maize straw were compared to no amendment (CK). Topsoil SOM was sequentially extracted with ultrapure water (UWE), solvent (TSE), base hydrolysis (BHY) and CuO oxidation (CUO) and molecular compounds in these extracted fractions were detected quantitatively by GC/MS. Compared to CK, SOC content was increased respectively by 12% under CS and CM, and by 36% under CB. Fraction abundance both of UWE and CUO was increased but that of TSE unchanged under CS and CM, while that of BHY unchanged under CS but increased by > 60% under CM and CB, respectively. Under CB relative to CS and CM, abundance of TSE and BHY fractions was greatly increased but that of UWE and CUO unchanged. Specifically, abundances of water-soluble monosaccharides, low molecular weight organic acids and lignin-derived phenols, especially cinnamyl-based monomers, were all enhanced under CS, but abundances of n-alkanols, fatty acids in free lipids and di-acids and hydroxyl fatty acids in bound lipids were decreased under both CS and CM. In contrast, CB amendment increased abundances of n-alkanols, alkanes and sterols in free lipids while enhanced molecular and functional group diversity of UWE and TSE fractions. Overall, short-term crop residue amendment altered the abundance and molecular diversity of OM mainly associated with short-lived UWE and labile TSE fractions and biochar enhanced preservation of plant derived molecules mainly in lipids. Thus, returning crop residue as biochar could be a sustainable approach to enhance not only SOM pool but molecular diversity also in agricultural soils.

Effect of amendment of biochar supplemented with Si on Cd mobility and rice uptake over three rice growing seasons in an acidic Cd-tainted paddy from central South China.

To examine the effect of rice straw biochar and the synergism with silicon on Cd immobilization, a Cd-contaminated acidic sandy loam paddy, polluted from emissions from industrial activity, was chosen in central South China. A field trial was conducted over three rice growing seasons during 2016-2017. Rice straw biochar (BR), produced by the pyrolysis of rice straw pellets at 450 °C, was amended at 10 t/ha (BR1), 20 t/ha (BR2), and supplemented with 0.75 t/ha sodium silicate (SS) at 10 t/ha, (BR1 + SS) and 20 t/ha (BR2 + SS), compared to the control without biochar and sodium silicate (BR0). BR supplemented with Si enhanced Cd soil immobilization and decreased Cd accumulation in rice plant within three rice seasons. Compared to BR0, BR + SS reduced Cd concentrations in grains by 19.5-73.7%, higher than that of 8.6-50.2% following BR. Cd bio-concentration factor of the root was reduced by an average of 54.6% from BR + SS and by 19.0% from BR, compared to BR0 in last two rice seasons. BR + SS significantly increased soil pH and available Si, and soil CaCl2-Cd was negatively related to soil available Si (p < 0.05). The synergistic effect of BR and Si induced liming effect and co-precipitation of Cd with Si compounds during the aging process of biochar. Thus, we suggest that an alkaline silicon supplementation is used as an amendment to BR, which could be used as a strategic approach for tackling Cd contamination in South China rice paddies.

Water-soluble mercury induced by organic amendments affected microbial community assemblage in mercury-polluted paddy soil.

Mercury (Hg) pollution or organic amendments (OA) may individually induce changes in the microbial community of paddy soils. However, little is known regarding the interaction of Hg and OA and the effect of different OA applications on the microbial community assemblage in Hg-polluted paddy soil. A soil incubation experiment was performed by applying three organic amendments (OA), namely a food-waste compost (FC), and its HA and FA, into an Hg-polluted paddy soil to examine the changes in the microbial community and merA/merB gene abundance. The results showed that the OA treatments promoted total (SOC) and dissolved organic carbon (DOC) in soils, which may harbor copiotrophic bacteria. The HA and FA treatments decreased microbial diversity and richness along with an increase of water-soluble Hg (WHg) through the complexation of DOC to Hg, which may be mainly attributed to the enhanced Hg biotoxicity to soil microbiome induced by the increased WHg under these two treatments. Additionally, the WHg enhancement also contributed to the increase of Hg-resistant bacteria and merA/merB gene abundance, and consequently, induced changes in the microbial community. These results indicated the interaction of Hg and different OA induced the variation of WHg fraction in paddy soil, which played a fundamental role in the distinct responses of the microbial community assemblage. Collectively, the application of FA and HA to Hg-polluted soil should be limited considering Hg risk to microbiome, and FC can be an alternative.

Molecular characterization of organic matter in two calcareous soils: the effects of an acid decarbonation treatment.

The molecular composition of soil organic matter (SOM) of two calcareous soils highly rich in carbonates was assessed before and after decarbonation by acid washing with HCl through 13C-CPMAS-NMR spectroscopy and off-line thermochemolysis coupled with gas chromatography and mass spectrometry (THM-GC-MS). The acidic treatment promoted a considerable concentration of organic matter in both soils, thus improving the identification of molecules otherwise not easily detectable. Decarbonation induced only a slight loss of soil organic carbon (SOC), corresponding to 1.4 and 2.7% for A and B soils respectively. The acidic treatment also led to an increase in the organic carbon/total nitrogen (OC/N) ratio in soil A, while an opposite variation was found for the second soil. Moreover, variations in the concentration and molecular distribution of specific compound classes present in SOM were caused by the acid washing of soils. As confirmed by both 13C-CPMAS-NMR and thermochemolysis results, the molecules most susceptible to the acid treatment were the carbohydrates, lignin monomers (G14 and G15), fatty acids (C18 saturated and unsaturated), fatty acids of microbial origin (C15, C17, and C19), hydroxy acids (C16, C18), and dioic acids (C18) which represent the components weakly bound to the organic matrix. Our findings not only showed the efficacy of the decarbonation treatment of calcareous soils with 3 N HCl, but also indicated how the acidic washing can improve the differentiation of soils on the basis of SOM molecular characteristics. Graphical abstract.

Organic amendments affect dissolved organic matter composition and mercury dissolution in pore waters of mercury-polluted paddy soil.

Organic amendments (OA) have been applied in many mercury (Hg)-polluted paddy soils to meet increasing food demands with scarce land resources. However, little is known on the effects of different OAs on Hg dissolution and the composition of dissolved organic matter (DOM) in soil pore waters, both of which may be associated with Hg mobility. Consequently, DOM composition and Hg release levels were investigated in soil pore waters after applying food waste compost (FC), fulvic acids (FA) and humic acids (HA) to Hg-polluted paddy soils. FA and HA treatments promoted increased abundances of humic- and fulvic-like substances in pore water DOM while FC amendment increased soluble microbial by-products. FA amendment and high levels of both HA and FC amendments greatly promoted Hg dissolution in pore waters that could be attributed to the complexation of Hg with different DOM components. However, among all DOM components, only UVA fulvic and visible humic-like substances were positively correlated with Hg release levels and total organic carbon. These results indicate that discrepant DOM compositions are induced by different OA. Further, these differences may be associated with differential Hg dissolution in pore waters. Consequently, FA amendment and high level of FC or HA amendments should be limited to reduce potential Hg release into pore waters.

HRMAS-NMR metabolomics of Aglianicone grapes pulp to evaluate terroir and vintage effects, and, as assessed by the electromagnetic induction (EMI) technique, spatial variability of vineyard soils.

Aglianicone is an autochthonous variety of black berried wine grape cultivated exclusively in southern Italy, but with limited information on the molecular composition of its grapes. HRMAS NMR spectroscopy was used to identify the primary metabolome of Aglianicone grape pulp from two different vineyards and the observed metabolomic changes reflected the local terroir conditions or harvesting years. Moreover, the berries pulp metabolome was related to Apparent Electrical Conductivity (ECa) maps deriving from on-site near-surface geophysical measurements of vineyard soils obtained by an electromagnetic induction (EMI) technique. Most of primary metabolites of Aglianicone grape varied systematically as a function of the spatial variability of soil properties as well as vineyards terroir and harvesting years. Therefore, the proposed approach that couples HRMAS NMR spectroscopy with ECa maps of vineyard soils represents an innovative support for wine producers who can select the best vine/soil combination to ensure the maximum wine quality.

Organic Carbon Sequestration in Soil Humic Substances As Affected by Application of Different Nitrogen Fertilizers in a Vegetable-Rotation Cropping System.

Little is known on the effect of application of different nitrogen (N) fertilizers on soil organic carbon (SOC) sequestration in soil humic substances (HS). We investigated HS molecular characteristics in an Orthic Acrisol, southwestern China, under 2-year field fertilization of a urea (U), a polymer-coated urea (PCU) and a biochar-coated urea (BCU) using 13C-CPMAS-NMR spectroscopy. Results showed that N fertilization promoted SOC sequestration into HS and favored alkyl-C and aromatic-C rather than O-alkyl-C and carbonyl-C for humic acids and humin in soil. Application of PCU and BCU may better enhance vegetable yield, SOC sequestration, and HS stability than the U application, which may benefit from longer time of N existence and higher total N in soil. Among the N treatments, BCU application mostly affected the compositions and stability of SOC in the HS for the OC input and prime effect of biochar for SOC transformation.

Replacing calcium with ammonium counterion in lignosulfonates from paper mills affects their molecular properties and bioactivity.

Lignosulfonates are important by-products of the paper industry and may be transformed into different commodities. We studied the molecular properties of ammonium (LS-AM) and calcium Lignosulfonates (LS-C) and evaluated their bioactivity towards the early development of maize plantlets. The FT-IR, 13C NMR and 1H-13C-HSQC-NMR spectra showed that the two lignosulfonates varied in hydroxyl, sulfonate and phenolic content, while DOSY-NMR spectroscopy suggested a similar diffusivity. High Performance Size Exclusion Chromatography (HPSEC) was used to simulate the effects of root-exuded acids and describe the conformational dynamics of both LS substrates in acidic aqueous solutions. This technique showed that LS-C was stabilized by the divalent Ca2+ counterion, thus showing a greater conformational stability than LS-AM, whose components could not be as efficiently aggregated by the monovalent NH4+ counter-ion. The plant bioassays revealed that LS-AM enhanced the elongation of the root system, whereas LS-C significantly increased both total and shoot plant weights. We concluded that the lignosulfonate bioactivity on plant growth depended on the applied concentrations, their molecular properties and conformational stability.

A molecular zoom into soil Humeome by a direct sequential chemical fractionation of soil.

A Humeomics sequential chemical fractionation coupled to advanced analytical identification was applied directly to soil for the first time. Humeomics extracted ~235% more soil organic carbon (SOC) than by the total alkaline extraction traditionally employed to solubilise soil humic molecules (soil Humeome). Seven fractions of either hydro- or organo-soluble components and a final unextractable humic residue were separated from soil. These materials enabled an unprecedented structural identification of solubilised heterogeneous humic molecules by combining NMR, GC-MS, and ESI-Orbitrap-MS. Identified molecules and their relative abundance were used to build up structure-based van Krevelen plots to show the specific contribution of each fraction to SOC. The stepwise isolation of mostly hydrophobic and unsaturated molecules of progressive structural complexity suggests that humic suprastructures in soil are arranged in multi-molecular layers. These comprised molecules either hydrophobically adsorbed on soil aluminosilicate surfaces in less stable fractions, or covalently bound in amorphous organo-iron complexes in more recalcitrant fractions. Moreover, most lipid molecules of the soil Humeome appeared to derive from plant polyesters rather than bacterial metabolism. An advanced understanding of soil humic molecular composition by Humeomics may enable control of the bio-organic dynamics and reactivity in soil.

The molecular properties of biochar carbon released in dilute acidic solution and its effects on maize seed germination.

It is not yet clear whether the carbon released from biochar in the soil solution stimulates biological activities. Soluble fractions (AQU) from wheat and maize biochars, whose molecular content was thoroughly characterized by FTIR, 13C and 1H NMR, and high-resolution ESI-IT-TOF-MS, were separated in dilute acidic solution to simulate soil rhizospheric conditions and their effects evaluated on maize seeds germination activity. Elongation of maize-seeds coleoptile was significantly promoted by maize biochar AQU, whereas it was inhibited by wheat biochar AQU. Both AQU fractions contained relatively small heterocyclic nitrogen compounds, whose structures were accounted by their spectroscopic properties. Point-of-Zero-Charge (PZC) values and van Krevelen plots of identified masses of soluble components suggested that the dissolved carbon from maize biochar behaved as humic-like supramolecular material capable to adhere to seedlings and deliver bioactive molecules. These findings contribute to understand the biostimulation potential of biochars from crop biomasses when applied in agricultural production.

Secondary Structures in a Freeze-Dried Lignite Humic Acid Fraction Caused by Hydrogen-Bonding of Acidic Protons with Aromatic Rings.

A lignite humic acid (HA) was separated from inorganic and non-HA impurities (i.e., aluminosilicates, metals) and fractionated by a combination of dialysis and XAD-8 resin. Fractionation revealed a more homogeneous structure of lignite HA. New and more specific structural information on the main lignite HA fraction is obtained by solid-state nuclear magnetic resonance (NMR) spectroscopy. Quantitative (13)C multiple cross-polarization (multiCP) NMR indicated oxidized phenyl propane structures derived from lignin. MultiCP experiments, conducted on potassium HA salts titrated to pH 10 and pH 12, revealed shifts consistent with carboxylate and phenolate formation, but structural changes associated with enolate formation from aromatic beta keto acids were not detected. Two-dimensional (1)H-(13)C heteronuclear correlation (2D HETCOR) NMR indicated aryl-aliphatic ketones, aliphatic and aromatic carboxyl groups, phenol, and methoxy phenyl ethers. Acidic protons from carboxyl groups in both the lignite HA fraction and a synthetic HA-like polycondensate were found to be hydrogen-bonded with electron-rich aromatic rings. Our results coupled with published infrared spectra provide evidence for the preferential hydrogen bonding of acidic hydrogens with electron-rich aromatic rings rather than adjacent carbonyl groups. These hydrogen-bonding interactions likely result from stereochemical arrangements in primary structures and folding.

Copolymerization of 2,4-dichlorophenol with humic substances by oxidative and photo-oxidative biomimetic catalysis.

We evaluated the catalytic activity of a water-soluble iron-porphyrin in an oxidative coupling reaction to form covalent bonds between 2,4-dichlorophenol (2,4-DCP) and humic molecules. The biomimetic catalysis in the presence of H2O2 was tested in the dark and in daylight, and changes in reaction products were revealed by high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) spectroscopy. In all conditions, iron-porphyrin was effective in promoting complete disappearance of 2,4-DCP, although catalyst activity was enhanced in daylight (with a maximum turnover number of 85.13). Further evidence of the occurred covalent coupling between 2,4-DCP and humic molecules was revealed by diffusion-ordered nuclear magnetic resonance (DOSY-NMR) spectroscopy that showed a reduced diffusivity of 2,4-DCP after the catalytic reaction. These findings indicate that iron-porphyrin is an efficient catalyst for the covalent binding of polyhalogenated phenols to humic molecules, thereby suggesting that the copolymerization reactions may become a useful technology to remediate soils and waters contaminated by halogenated phenols and their analogues.