Synthesis and properties of β-d-glucosidasephenolic copolymers as analogues of soil humic-enzyme complexes

Several β-d-glucosidase-phenolic copolymers were synthesized and three examined in detail: those containing l-tyrosine, pyrogallol and resorcinol. These copolymers were similar to naturallyoccurring soil humic-enzyme complexes in many ways: E₄/E₆ ratios, C, H, N and S content and IR spectra. The enzyme activity of the copolymers showed varying degrees of resistance to proteolysis, organic solvents, and storage at high temperatures. All immobilized enzymes had increased K_m values and decreased V_max values in comparison with soluble β-d-glucosidase (9.3 mM, 190μmol p-nitrophenol mg^?1h^?1); the β-d-glucosidase-resorcinol copolymer was the most active (10.5 mM, 104μmol p-nitrophenol mg^?1h^?1). β-d-Glucosidase activity was completely resistant to protease when the copolymer was fixed to bentonite clay but V_max values were reduced further (e.g. β-d-glucosidase-resorcinol-bentonite complex, 58.5μmol p-nitrophenol mg^?1 h^?1). On addition to soil, soluble β-d-glucosidase was rapidly inactivated (38% loss in 3 days, 80% loss in 21 days) whereas β-d-glucosidase-resorcinol/pyrogallol and β-d-glucosidase-L-tyrosine copolymers were comparatively stable (no loss in 9 days, 25% and 44% loss in 21 days). It is suggested that the copolymerization of enzyme during humic matter formation is a major factor leading to the stabilisation of soil enzymes and that adsorption and entrapment are comparatively insignificant.     

Use of 0·1 m pyrophosphate to extract urease from a podzol

Sodium pyrophosphatc (0·1 m) at pH 7.1 and 37°C extracted a significant fraction of urease from a podzol. Maximum extraction values were obtained after 18 h. The yields of soil organic matter and urease activity during the extraction show a different pattern: the extraction of non-specific organic matter precedes and may facilitate the following extraction of an active urease organo-complex. The urease extracted by pyrophosphate is about 30 40 per cent of the total urease activity, as shown by plotting the urease activity against the population changes of ureolytic microorganisms, both in the original and extracted soil. The number of ureolytic microorganisms is unaffected by pyrophosphate, and the extracted urease is assumed to be extracellular.     

Mineralization of dissolved organic carbon in mineral soil solution of two forest soils

Dissolved organic carbon (DOC) constitutes an important carbon input flux to forested mineral soils. Seepage from mineral subsoils contains only small amounts of DOC because of mineralization, sorption or the formation of particulate organic matter (POM). However, the relation between these processes is largely unknown. Therefore, the objective of this study was to quantify the mineralization of DOC from different depths of forest soils, and to determine degradation rate constants for rapidly and slowly degradable DOC pools. Mineralization of DOC and formation of POM in mineral soil solution from two forested sites in northern Bavaria (Germany) were quantified in a 97 days laboratory incubation experiment. Furthermore, spectroscopic properties such as specific UV absorption and a humification index derived from fluorescence emission spectrometry were measured before and after incubation. DOC in all samples turned out to belong mainly to the stable DOC pool (> 95 %) with half‐lives ranging from years to decades. Spectroscopic properties were not suitable to predict the mineralization of DOC from mineral soils. However, together with data on DOC from the forest floor and long‐term data on DOC concentrations in the field they helped to identify the processes involved in C sequestration in mineral subsoils. Mineralization, formation of POM, and probably sorption seem all to be responsible for maintaining low concentrations of DOC in the upper mineral soil. DOC below the upper mineral soil is highly resistant to mineralization, and thus the further decrease of DOC concentrations in the subsoil as observed under field conditions cannot be attributed to mineralization. Our results suggest that sorption and to some minor extent the formation of POM may be responsible for C sequestration in the subsoil.     

The influence of soluble carbon and fertilizer nitrogen on nitric oxide and nitrous oxide emissions from two contrasting agricultural soils

Contradictory effects of simultaneous available organic C and N sources on nitrous oxide (N₂O), carbon dioxide (CO₂) and nitric oxide (NO) fluxes are reported in the literature. In order to clarify this controversy, laboratory experiments were conduced on two different soils, a semiarid arable soil from Spain (soil I, pH=7.5, 0.8%C) and a grassland soil from Scotland (soil II, pH=5.5, 4.1%C). Soils were incubated at two different moisture contents, at a water filled pore space (WFPS) of 90% and 40%. Ammonium sulphate, added at rates equivalent to 200 and 50 kg N ha^?1, stimulated N₂O and NO emissions in both soils. Under wet conditions (90% WFPS), at high and low rates of N additions, cumulative N₂O emissions increased by 250.7 and 8.1 ng N₂O–N g^?1 in comparison to the control, respectively, in soil I and by 472.2 and 2.1 ng N₂O–N g^?1, respectively, in soil II. NO emissions only significantly increased in soil I at the high N application rate with and without glucose addition and at both 40% and 90% WFPS. In both soils additions of glucose together with the high N application rate (200 kg N ha^?1) reduced cumulative N₂O and NO emissions by 94% and 55% in soil I, and by 46% and 66% in soil II, respectively. These differences can be explained by differences in soil properties, including pH, soil mineral N and total and dissolved organic carbon content. It is speculated that nitrifier denitrification was the main source of NO and N₂O in the C-poor Spanish soil, and coupled nitrification–denitrification in the C-rich Scottish soil.     

The influence of time, storage temperature, and substrate age on potential soil enzyme activity in acidic forest soils using MUB-linked substrates and l-DOPA

The purpose of this experiment was to evaluate whether soil storage and processing methods significantly influence measurements of potential in situ enzyme activity in acidic forest soils. More specifically, the objectives were to determine if: (1) duration and temperature of soil storage; (2) duration of soil slurry in buffer; and (3) age of model substrates significantly influence the activity of six commonly measured soil extracellular enzymes using methylumbelliferone (MUB)-linked substrates and l-dihydroxyphenylalanine (l-DOPA). Soil collected and analyzed for enzyme activity within 2 h was considered the best measure of potential in situ enzyme activity and the benchmark for all statistical comparisons. Sub-samples of the same soil were stored at either 4 °C or −20 °C. In addition to the temperature manipulation, soils experienced two more experimental treatments. First, enzyme activity was analyzed 2, 7, 14, and 21 days after collection. Second, MUB-linked substrate was added immediately (i.e. <20 min) or 2 h after mixing soil with buffer. Enzyme activity of soil stored at 4 °C was not significantly different from soil stored at −20 °C. The duration of soil storage was minimal for β-glucosidase, β-xylosidase, and peroxidase activity. N-acetyl-glucosaminidase (NAGase), phosphatase, and phenol oxidase activity appeared to change the most when compared to fresh soils, but the direction of change varied. Likewise, the activities of these enzymes were most sensitive to extended time in buffer. Fluorometric MUB and MUB-linked substrates generally had a 3-day shelf life before they start to significantly suppress reported activities when kept at 4 °C. These findings suggest that the manner in which acidic forest soils are stored and processed are site and enzyme specific and should not initially be trivialized when conducting enzyme assays focusing on NAGase, phosphatase, and phenol oxidase. The activities of β-glucosidase, β-xylosidase, and peroxidase are insensitive to storage and processing methods.     

Mineralization of carbon and nitrogen from fresh and anaerobically stored sheep manure in soils of different texture

A sandy loam soil was mixed with three different amounts of quartz sand and incubated with (¹⁵NH₄)₂SO₄ (60 g N g^-1 soil) and fresh or anaerobically stored sheep manure (60 g g^-1 soil). The mineralization-immobilization of N and the mineralization of C were studied during 84 days of incubation at 20°C. After 7 days, the amount of unlabelled inorganic N in the manure-treated soils was 6–10 g N g^-1 soil higher than in soils amended with only (¹⁵NH₄)₂SO₄. However, due to immobilization of labelled inorganic N, the resulting net mineralization of N from manure was insignificant or slightly negative in the three soil-sand mixtures (100% soil+0% quartz sand; 50% soil+50% quartz sand; 25% soil+75% quartz sand). After 84 days, the cumulative CO₂ evolution and the net mineralization of N from the fresh manure were highest in the soil-sand mixutre with the lowest clay content (4% clay); 28% fo the manure C and 18% of the manure N were net mineralized. There was no significant difference between the soil-sand mixtures containing 8% and 16% clay, in which 24% of the manure C and -1% to 4% of the manure N were net mineralized. The higher net mineralization of N in the soil-sand mixture with the lowest clay content was probably caused by a higher remineralization of immobilized N in this soil-sand mixture. Anaerobic storage of the manure reduced the CO₂ evolution rates from the manure C in the three soil-sand mixtures during the initial weeks of decomposition. However, there was no effect of storage on net mineralization of N at the end of the incubation period. Hence, there was no apparent relationship between net mineralization of manure N and C.     

Mineralization of carbon and nitrogen from cowpea leaves decomposing in soils with different levels of microbial biomass

Soils with greater levels of microbial biomass may be able to release nutrients more rapidly from applied plant material. We tested the hypothesis that the indigenous soil microbial biomass affects the rate of decomposition of added green manure. Cowpea (Vigna unguiculata L.) Walp.] leaves were added to four soils with widely differing microbial biomass C levels. C and N mineralization of the added plant material was followed during incubation at 30°C for 60 days. Low levels of soil microbial biomass resulted in an initially slower rate of decomposition of soil-incorporated green manure. The microbial biomass appeared to adjust rapidly to the new substrate, so that at 60 days of incubation the cumulative C loss and net N mineralization from decomposing cowpea leaves were not significantly affected by the level of the indigenous soil microbial biomass.     

Rhizodeposits of Trifolium pratense and Lolium perenne: their comparative effects on 2,4-D mineralization in two contrasting soils

Rhizosphere enhanced biodegradation of organic pollutants has been reported frequently and a stimulatory role for specific components of rhizodeposits postulated. As rhizodeposit composition is a function of plant species and soil type, we compared the effect of Lolium perenne and Trifolium pratense grown in two different soils (a sandy silt loam: pH 4, 2.8% OC, no previous 2,4-D exposure and a silt loam: pH 6.5, 4.3% OC, previous 2,4-D exposure) on the mineralization of the herbicide 2,4-D (2,4-dichlorophenoxyacetic acid). We investigated the relationship of mineralization kinetics to dehydrogenase activity, most probable number of 2,4-D degraders (MPN_2,4-D) and 2,4-D degrader composition (using sequence analysis of the gene encoding α-ketoglutarate/2,4-D dioxygenase (tfdA)). There were significant (P<0.01) plant-soil interaction effects on MPN_2,4-D and 2,4-D mineralization kinetics (e.g. T. pratense rhizodeposits enhanced the maximum mineralization rate by 30% in the acid sandy silt loam soil, but not in the neutral silt loam soil). Differences in mineralization kinetics could not be ascribed to 2,4-D degrader composition as both soils had tfdA sequences which clustered with tfdAs representative of two distinct classes of 2,4-D degrader: canonical R. eutropha JMP134-like and oligotrophic α-proteobacterial-like. Other explanations for the differential rhizodeposit effect between soils and plants (e.g. nutrient competition effects) are discussed. Our findings stress that complexity of soil-plant-microbe interactions in the rhizosphere make the occurrence and extent of rhizosphere-enhanced xenobiotic degradation difficult to predict.     

Retention and hydrolysable fraction of atmospherically deposited nitrogen in two contrasting forest soils in Switzerland

Nitrogen (N) from atmospheric deposition has been shown to be mainly retained in the organic soil layers of temperate forest ecosystems, but the mechanisms and the physico‐chemical fractions involved are still poorly defined. We performed a hot‐acid hydrolysis on ¹⁵N‐labelled soil samples collected 1 week, 3 months and 1 year following a single in situ application of either ¹⁵NO₃⁻ or ¹⁵NH₄⁺ in two montane forest ecosystems in Switzerland: Grandvillard (beech forest on a calcareous, well‐drained soil, 650 m above sea level) and Alptal (spruce forest on hydromorphic soil, 1200 m above sea level). After ¹⁵NH₄⁺ application, recovery rates in the soil were smaller in Alptal than in Grandvillard through a large rate of absorption by mosses. At both sites, the organic soil layers retained most of the tracers at all three sampling times between 1 week and 1 year. In Grandvillard, the hydrolysable fraction (hydrolysable N : total N) of ¹⁵N was on average 79% and thus similar to the hydrolysable fraction of native N. This similarity is probably because of the rapid incorporation of N into organic molecules, followed by stabilization of the recalcitrant N pool through organo‐mineral bonds with soil minerals. In Alptal, the ¹⁵N hydrolysable fraction was greater than that of native N, particularly after ¹⁵NH₄⁺ application (¹⁵N, 84%; native N, 72%). At both sites, ¹⁵N and the fraction of hydrolysable native N remained constant between 1 week and 1 year. This shows that both the recalcitrant and the hydrolysable pools are stable in the mid‐ to long‐term. We present arguments indicating that biological recycling through microbes and plants contributes to the stability of the hydrolysable N fraction.     

Biochar differentially affects the cycling and partitioning of low molecular weight carbon in contrasting soils

Biochar application to soils has received much attention due to the potential for dual benefits of improved fertility and carbon (C) sequestration. Whilst its effect on C and nitrogen (N) cycling in soils has been investigated previously, this has usually either focussed on the bulk soil organic matter, or a single compound such as glucose. Five low molecular weight dissolved organic C (LMWDOC) substrates (three sugars, one amino acid, one organic acid) were selected for a ¹⁴C-CLPP experiment from which turnover rate (t_1/2) and immediate carbon use efficiency (CUE) of the substrate were estimated. We demonstrated that whilst soil type had the greatest effect on soil microbial function, the addition of biochar also influenced microbial turnover and CUE of the substrates, most notably in the lowest fertility soil. We also identified that the relationship between turnover and CUE of the five substrates differed substantially, and the effect of biochar and soil type was more pronounced in the amino acid than the organic acid. This effect tended to be greatest in biochars produced at 450 °C, and less pronounced with the addition of biochars produced at 550 °C, though these trends were not consistent for all compounds in all soils tested. We conclude biochars and soils interact to manifest non-systematic differences in turnover rates of LMWDOCs, and thus a variety of mechanisms are likely responsible for this observation. As these compounds are most commonly found in the rhizosphere and can contribute a significant portion of photosynthetically-fixed C, and plant roots have been observed to grow preferentially around biochar particles, it is apparent that biochar may significantly affect the flow of LMWDOC through the microbial community in soils.     

Environmental impacts of abandoned dredged soils and sediments Available options for their handling, restoration and rehabilitation

Aim and background  

In the process of creating safe navigable waterways for oil exploitation, the companies operating in the Niger Delta generate tons of sulfidic spoils. These are often deposited over bank, mostly upon fringing mangroves, and abandoned. This leads to a myriad of environmental problems. The extent of these impacts is not exactly known, but was inferred from the activities of oil companies operating in the area. This paper describes the impacts following the disturbance of sulfidic sediment through dredging and by subsequent poor spoil management practices. Environmental impacts of exposed and abandoned sulfidic sediments. The practice of dumping and abandoning sulfidic dredged spoils along canal banks triggers a series of environmental problems leading to extreme acidification, heavy metal pollution, and general habitat degradation which prevent the re-colonization of the sites by native species. The resultant spoil dumps remain bare for several years and they become colonized by invasive species later. Later still they may become attractive to the local population as sites for houses, fishing camps and home gardens, which is nevertheless regarded as a positive impact.     

Earthworm burrowing activity of two non-Lumbricidae earthworm species incubated in soils with contrasting organic carbon content (Vertisol vs. Ultisol)

The aim of this study was to investigate the burrowing activity of two earthworm species: the endogeic Drawida sinica and one undescribed Amynthas species incubated in Vertisol and Ultisol presenting different soil organic C content. Because of their contrasting feeding behaviours, we hypothesised that soil type would have a bigger influence on the burrowing activity of the endogeic than the anecic species. Repacked soil columns inoculated with earthworms for 30 days were scanned using X-ray tomography and the compiled images used to characterise the burrow systems. After scanning, the saturated hydraulic conductivity (K _sat) was also measured. The Amynthas species burrows were less numerous (30 vs. 180), more vertically oriented (57 vs. 37°), more connected from the surface to the bottom of the columns (73 vs. 5 cm³) and had a higher global connectivity index (83 vs. 28%) than those of D. sinica. The K _sat was threefold faster in columns incubated with Amynthas and was linked to the volume of percolating burrows (R ² = 0.81). The soil type did not influence Amynthas burrow characteristics. In contrast, there were 30% more D. sinica burrows in the Vertisol than in the Ultisol while other burrow characteristics were not affected. This result suggests that these burrows were more refilled with casts leading to shorter and discontinuous burrows. The K _sat was negatively related to the number of burrows (R ² = 0.44) but was not statistically different between the Vertisol and the Ultisol, suggesting a constant impact of this species on the K _sat. We found that a decrease in the amount of soil organic C by 50% had only a small influence on earthworm burrowing activity and no effect on the K _sat.     

pH regulation of carbon and nitrogen dynamics in two agricultural soils

Soil pH is often hypothesized to be a major factor regulating organic matter turnover and inorganic nitrogen production in agricultural soils. The aim of this study was to critically test the relationship between soil pH and rates of C and N cycling, and dissolved organic nitrogen (DON), in two long-term field experiments in which pH had been manipulated (Rothamsted silty clay loam, pH 3.5-6.8; Woburn sandy loam, pH 3.4-6.3). While alteration of pH for 37 years significantly affected crop production, it had no significant effect on total soil C and N or indigenous mineral N levels. This implies that at steady state, increased organic matter inputs to the soil are balanced by increased outputs of CO₂. This is supported by the positive correlation between both plant productivity and intrinsic microbial respiration with soil pH. In addition, soil microbial biomass C and N, and nitrification were also significantly positively correlated with soil pH. Measurements of respiration following addition of urea and amino acids showed a significant decline in CO₂ evolution with increasing soil acidity, whilst glucose mineralization showed no response to pH. In conclusion, it appears that changes in soil pH significantly affect soil microbial activity and the rate of soil C and N cycling. The evidence suggests that this response is partially indirect, being primarily linked to pH induced changes in net primary production and the availability of substrates. In addition, enhanced soil acidity may also act directly on the functioning of the microbial community itself.     

Long-term impact of reduced tillage and residue management on soil carbon stabilization: Implications for conservation agriculture on contrasting soils

Residue retention and reduced tillage are both conservation agricultural management options that may enhance soil organic carbon (SOC) stabilization in tropical soils. Therefore, we evaluated the effects of long-term tillage and residue management on SOC dynamics in a Chromic Luvisol (red clay soil) and Areni-Gleyic Luvisol (sandy soil) in Zimbabwe. At the time of sampling the soils had been under conventional tillage (CT), mulch ripping (MR), clean ripping (CR) and tied ridging (TR) for 9 years. Soil was fully dispersed and separated into 212–2000 μm (coarse sand), 53–212 μm (fine sand), 20–53 μm (coarse silt), 5–20 μm (fine silt) and 0–5 μm (clay) size fractions. The whole soil and size fractions were analyzed for C content. Conventional tillage treatments had the least amount of SOC, with 14.9 mg C g⁻¹ soil and 4.2 mg C g⁻¹ soil for the red clay and sandy soils, respectively. The highest SOC content was 6.8 mg C g⁻¹ soil in the sandy soil under MR, whereas for the red clay soil, TR had the highest SOC content of 20.4 mg C g⁻¹ soil. Organic C in the size fractions increased with decreasing size of the fractions. In both soils, the smallest response to management was observed in the clay size fractions, confirming that this size fraction is the most stable. The coarse sand-size fraction was most responsive to management in the sandy soil where MR had 42% more organic C than CR, suggesting that SOC contents of this fraction are predominantly controlled by amounts of C input. In contrast, the fine sand fraction was the most responsive fraction in the red clay soil with a 66% greater C content in the TR than CT. This result suggests that tillage disturbance is the dominant factor reducing C stabilization in a clayey soil, probably by reducing C stabilization within microaggregates. In conclusion, developing viable conservation agriculture practices to optimize SOC contents and long-term agroecosystem sustainability should prioritize the maintenance of C inputs (e.g. residue retention) to coarse textured soils, but should focus on the reduction of SOC decomposition (e.g. through reduced tillage) in fine textured soils.