Spatial distribution of glomalin-related soil protein and its relationships with root mycorrhization,soil aggregates,carbohydrates, activity of protease and β-glucosidase in the rhizosphere of Citrus unshiu

Relationships between the spatial distributions of glomalin-related soil protein (GRSP) and soil aggregates, carbohydrates or relevant enzymes are poorly studied. We found that two categories of GRSP, the easily extractable Bradford-reactive soil protein (EE-BRSP) and total BRSP (T-BRSP), respectively ranged between 0.3–0.6 and 0.5–0.8 mg/g DW soil, and these two BRSPs decreased with the increase of soil depth (0–40 cm) in the rhizosphere of a 22-year-old Citrus unshiu orchard. Both EE-BRSP and T-BRSP were significantly positively correlated with mycorrhization, 0.25–0.50 mm soil water-stable aggregates, water-extractable or hydrolyzable carbohydrates, and β-glucosidase, but significantly negatively correlated with protease. Our results demonstrate that the spatial distribution of GRSP is significantly affected by mycorrhization, soil carbohydrate, β-glucosidase and protease.     

A method for the determination of β-glucosidase activity in soil

A method is described for the rapid and simple assay of soil β-glucosidase activity. It involves colorimetric estimation of ρ-nitrophenol released by β-glucosidase activity when soil is incubated in McIlvaine buffer (pH 4.8) with ρnitrophenyl βd-glucoside and toluene at 30°C for 1 hr. The method has been applied to three different soils. The range of β-glucosidase activity in cultivated soils was from 10.1 to 15.2 mµ mole per min per gram of dried soil. K_m value for ρ-nitrophenyl β-d-glucoside was 3.3 × 10^-4 M. Optimum pH was 4.8.     

Extraction of β-glucosidase activity from PEA field soil

Brown compounds with β-glucosidase activity were extracted from a cultivated soil in 0.1 m phosphate buffer (pH 7), 0.3 m KCl and 10 mm EDTA. A curvilinear relationship of the Langmuir type was observed between the solution volume and β-glucosidase activity of the extract. The results indicated the β-glucosidase was extracellular and was adsorbed on the surfaces of soil particles by ionic bonds. The preparation was treated with protamine sulfate and its enzymatic properties were investigated. The activity of the preparation was optimal at pH 5.4 and was lost after 10 min at 80°C. The extract hydrolyzed p-nitrophenyl β-glucoside(100), phenyl β-glucoside(19), salicin(8), amygdalin(33), cellobiose(52) and gentiobiose(55) (relative activities shown in parentheses). But, the extract had no effect on methyl β-glucoside and phlorizin. The substrate specificity and optimum pH of the enzymatic activity of the soil extract was similar to those of β-glucosidases from various fungi.     

Hydrolysis of cellobiose by β-glucosidase in the presence of soil minerals - Interactions at solid-liquid interfaces and effects on enzyme activity levels

Extracellular enzymatic activities in soils are essential for the cycling of organic matter. These activities take place in multiphase environments where solid phases profoundly affect biocatalytic activities. Aspergillus niger is ubiquitous in soils; its β-glucosidase plays an important role in the degradation of cellulose, and therefore in the global carbon cycle and in the turnover of soil organic matter. However, the information on the interactions of this protein with soil minerals is very limited, and even less is known about their consequences for the hydrolysis of the natural substrate cellobiose. We therefore characterised the sorptive interactions of this enzyme with the soil minerals montmorillonite, kaolinite and goethite and quantified the resulting changes in the hydrolysis rate of cellobiose. Fractions of adsorbed protein, and the resulting catalytic activity loss, were lower for montmorillonite than for kaolinite and goethite at given experimental conditions; adsorption was 9.7 ± 7.3% for montmorillonite, 70.3 ± 3.1% for kaolinite and 71.4 ± 1.8% for goethite, respectively. Adsorption of the protein to the minerals caused a total decrease in the catalytic activity of 18.8 ± 3.4% for kaolinite and 17.9 ± 4.7% for goethite whereas it was not significant for montmorillonite. The average catalytic activity lost by the pool of adsorbed molecules was 26.8% for kaolinite and 25.0% for goethite. Both the amount of adsorbed protein and the resulting loss of catalytic activity were found to be independent of the specific surface areas yet were influenced by the electrical properties of the mineral surfaces. Under the experimental conditions, montmorillonite and kaolinite are negatively charged whereas goethite is positively charged. However, because of the adsorption of phosphate anions from the buffer, a charge reversal took place at the surface of goethite. This was confirmed by zeta (ζ)-potential measurements in phosphate buffer, revealing negative values for all the tested minerals. Indeed goethite interacted with the enzyme as a negatively charged surface: the amount of adsorbed protein and the resulting catalytic activity loss were very similar to those of kaolinite. Our results show that, even if an important fraction of β-glucosidase is adsorbed to the minerals, the catalytic activity is largely retained. We suggest that this strong activity retention in presence of soil minerals results from a selective pressure on A. niger, which benefits from the activity of the adsorbed, and thus stabilized, enzyme pool.     

Effects of copper on the decomposition of plant residues,microbial biomass,and β-glucosidase activity in soils

Abstract

Two types of soils (Brown Lowland soil and Ando soil), which were artificially enriched with different amounts of Cu, were incubated with or without pulverized orchard grass for 12 weeks at 25°C. For both soils with and without orchard grass amendment, the amount of CO₂ evolved over the 12-week period of incubation decreased by the enrichment with Cu at a concentration exceeding 1,000 mg kg^?1 soil. The decrease of the mineralization of added orchard grass in the Cu-enriched soil was conspicuous especially during the initial period of incubation. The amount of microbial biomass C at the end of the incubation was significantly reduced by the Cu enrichment regardless of the amendment with orchard grass. The relative decrease of the soil microbial biomass was much greater than that of the soil respiration. The amount of biomass C was negatively correlated with the amount of 0.1 M CaCl₂-extractable Cu as a logarithmic function. On the other hand, the β-glucosidase activity at the end of the incubation was not significantly affected by the presence of Cu in the soils without orchard grass amendment and increased with the increase in the amount of enriched Cu in the orchard grass-amended soils.     

Characterization and antioxidant activity of the complex of tea polyphenols and oat β-glucan

Few data are available about the effects of complexation of polyphenols with polysaccharide on their bioavailability. The complex of tea polyphenols (TP) with oat β-glucan was characterized by ultraviolet-visible spectrometry, Fourier transform infrared spectrometry, differential scanning calorimetry, atomic force microscopy, and solid-state (13)C NMR spectroscopy. The results indicated that the bonds which governed the interaction between TP and oat β-glucan were strong hydrogen bonds. The in vitro antioxidant activity of TP, β-glucan, their complex, and physical mixture was assessed using four systems, namely, DPPH(?), OH(?), and O(2)(?-) scavenging activities and reducing power. The complexation and blending of TP and β-glucan exhibited different impacts on the index of in vitro and in vivo antioxidant capacities. In the concentration range of 0.5-2.5 mg mL(-1), the complex had highest O(2)(?-) scavenging activity, whereas the highest OH(?) scavenging activity was found with the physical mixture. For antioxidant testing in vivo, there was no significant difference between the complex and the physical mixture in terms of glutathione peroxidase activity and levels of malondialdehyde and total antioxidant capacity in serums. However, the complex exhibited much higher activities of superoxide dismutase and glutathione peroxidase in livers than the physical mixture. The present study provided a deeper understanding of the influence of molecular interaction between TP and oat β-glucan on their antioxidant activities.     

Spatial variation of nitrate–N and related soil properties at the plot-scale

Neglecting the spatial variation in soil nutrient status may result in unused yield potential and in environmental damage. Site-specific management has been suggested to reduce inappropriate fertilization that can adversely affect soil, ground and surface water. Decision criteria for determining variable-rate nitrogen fertilization are, however, lacking. This paper analyses the spatial variation of nitrate nitrogen (NO₃–N) and soil properties related to the N cycle at the plot-scale. Three 50×50 m plots were sampled in nested sampling designs of varying complexities. Classical statistics revealed a characteristic ranking in the variability of soil properties. Geostatistical analysis of the NO₃–N data from two plots showed that the small-scale variation found in one small subgrid was not typical for the small-scale variation in the entire plot, indicating bias in the sampling design. A trend component was found in the NO₃–N data and, consequently, the minimal requirement for the regionalized variable theory was not fulfilled. Problems due to design were overcome with a more complex nested sampling at the third plot. However, the spherical model fitted to the NO₃–N data of the first year explained only 21% of the total variance, whereas a pure nugget effect was observed in the second year. The water content data also showed a low structural variance, which was different in the two years. In contrast, two thirds of the variance of total carbon (C_t) and total nitrogen (N_t) could be explained by the fitted models. Seasonal variations, such as varying duration of snow cover, and extrinsic management effects, such as growing of a cover crop, may have contributed to the observed differences in variability between the years. Due to the low proportion of structural variance and the observation that spatial distribution was not stable with time, geostatistical analysis of NO₃–N and water contents data added only little information to classical statistical analysis. However, geostatistical analysis of total C and N contents provided a useful means to calculate spatial distribution patterns of these properties.     

Contact angles at the water–air interface of hydrocarbon-contaminated soils and clay minerals

Contact angles at the water–air interface have been measured for triturated preparations of clays and soils in order to assess changes in their hydrophobic properties under the effect of oil hydrocarbons. Tasks have been to determine the dynamics of contact angle under soil wetting conditions and to reveal the effect of chemical removal of organic matter from soils on the hydrophilicity of preparations. The potentialities of static and dynamic drop tests for assessing the hydrophilic–hydrophobic properties of soils have been estimated. Clays (kaolinite, gumbrine, and argillite) have been investigated, as well as plow horizons of soils from the Republic of Tatarstan: heavy loamy leached chernozem, medium loamy dark gray forest soil, and light loamy soddy-calcareous soil. The soils have been contaminated with raw oil and kerosene at rates of 0.1–3 wt %. In the uncontaminated and contaminated chernozem, capillary water capacity has been maintained for 250 days. The contact angles have been found to depend on the degree of dispersion of powdered preparation, the main type of clay minerals in the soil, the presence and amount of oxidation-resistant soil organic matter, and the soil–water contact time. Characteristic parameters of mathematical models for drop behavior on triturated preparations have been calculated. Contamination with hydrocarbons has resulted in a reliable increase in the contact angles of soil preparations. The hydrophobization of soil surface in chernozem is more active than in soils poorer in organic matter. The complete restoration of the hydrophilic properties of soils after hydrocarbon contamination is due to the oxidation of easily oxidizable organic matter at the low content of humus, or to wetting during several months in the absence of the mazut fraction.     

Isoelectric focusing of β-glucosidase humic-bound activity in semi-arid Mediterranean soils under management practices

Changes in β-glucosidase enzyme–humic complexes and conventional parameters (pH, total organic C, total N, water-soluble C, and bulk density) were studied in an almond-cropped soil prone to erosion under a rehabilitation practice. The experimental plan included three soil slopes (0%, 2%, and 6%) and two type of fertilization (organic and mineral), with sampling of rhizosphere and inter-row soils. The enzyme humic complexes were extracted by pyrophosphate, purified by ultrafiltration of the organic extracts on molecular mass exclusion membranes (mol wt > 10⁴) and fractionated by isoelectric focusing technique (IEF). The IEF on polyacrylamide rod gels with a restricted pH gradient ranging between 6.0 and 4.0 gave five humic bands on the basis of the little differences of their electric charges (pI). Under both organic and mineral fertilization, β-glucosidase activity bound to the fractionated humic substances, especially in the pH range 4.5–4.2 of the rhizosphere soil, was higher than that of the inter-row soil. This also occurred in 6% slope where the enzyme activity was lower than in soil with lower slopes. The higher number of the β-glucosidase active humic bands in rhizosphere than inter-row soil, particularly for the 0% slope, may be due to the presence of humic molecules capable of preserving the enzyme molecules in the active form, other than to the higher microbial activity synthesizing and releasing the tested enzymes.     

Effect of the substrate concentration and water activity on the yield and rate of the transfer reaction of β-galactosidase from Bacillus circulans

Prebiotic galactosyl oligosaccharides (GOS) are produced from lactose by the enzyme β-galactosidase. It is widely reported that the highest GOS levels are achieved when the initial lactose concentration is as high as possible; however, little evidence has been presented to explain this phenomenon. Using a system composed of the commercial β-galactosidase derived from Bacillus circulans known as Biolacta FN5, lactose and sucrose, the relative contribution of water activity, and substrate availability were assessed. Oligosaccharide levels did not appear to be affected by changes in water activity between 1.0 and 0.77 at a constant lactose concentration. The maximum oligosaccharide concentration increased at higher initial concentrations of lactose and sucrose, while initial reaction rates for transfer increased but remained constant for hydrolysis. This suggests that the high oligosaccharide levels achieved at the raised initial saccharide concentration are due to increases in reactions that form oligosaccharides rather than decreases in concurrent reactions, which degrade oligosaccharides. There were different effects from changing the initial concentration of lactose compared to sucrose, suggesting that the ability of lactose to act as a donor saccharide may be more important for increasing maximum oligosaccharide concentrations than the combined ability of both saccharides to act as galactosyl acceptors.     

Differentiation and geographical distribution of β-amylase isozyme in barley

This study reports on the investigation of the polymorphism and geographical distribution of -amylase isozymes by isoelectric-focusing (IEF) analysis in samples of world barley. The isozyme pattern of high thermostability type A and low thermostability type C varieties was restricted in isozyme type II. However, the isozyme pattern of the middle thermostability type B varieties was polymorphic. The -amylase isozyme of 1,554 lines of the middle thermostability group was investigated next. Besides types I and II, new isozyme types III and IV were observed. Most lines were grouped into type II in all growing areas, except for Europe where type I and type II were even in number. Types III and IV were observed in only one line in India and Afghanistan, respectively. In Northern Europe, especially in Denmark, type I was major. It was concluded that a mutant IEF isozyme type I originated from the type II prototype of the thermostability B group in Northern Europe.     

Effects of three biochar types on activity of β-glucosidase enzyme in two agricultural soils of different textures

ABSTRACT

Biochar has attracted significant attention due to the long-lasting nature, and prominent influence on soil characteristics. This study was conducted to evaluate changes in the activity of β-glucosidase enzyme (BG) in loamy and sandy loam texture soils following two winter wheat growing seasons. The experimental design was a randomized complete block with three replicates and four treatments. The treatments were two soils, three biochars (rice husk, corn cob and bean harvest residue), five biochar rates (BR) and five levels of mineral fertilizers (FR) or dairy effluent (DE). The fertilizers were applied at the beginning of each season, while biochars were applied only at the beginning of experiment. Soil samples were collected following the second season and analyzed for the BG activity. Addition of biochars reduced the BG activity and the decline was higher in sandy loam compared to loamy soils. Negative effect of biochar to BG activity was greater at the highest BR (3.0%) than the lower BR. Fertilizer additions along with DE biochar had significant effect on BG activity that increased with higher FR. Given the importance of BG activity in soil organic matter decomposition, biochar application can be considered a sustainable way of increase in carbon sequestration.     

Effects of fine bubble aeration at the sediment‒water interface on distributions of organic phosphorus fractions and related microbial activity in a heavily urban river

Journal of Soils and Sediments - Effects of aeration on distribution and release potential of organic phosphorus in sediments are of great significance. The aim of this study was to investigate...     

Effects of wood char and activated carbon on the hydrolysis of cellobiose by β-glucosidase from Aspergillus niger

Biochar amendments to soils have been suggested as a strategy to sequester carbon and therefore mitigate global climate change. The enrichment of soils with charred materials also increases their fertility. This fertilising effect of biochar may be caused by various mechanisms; an acceleration of nutrient cycling has been suggested as one such mechanism. The rate-limiting step in nutrient cycling is thought to be the extracellular enzymatic attack on biological macromolecules. In this study, therefore, the effects of chestnut wood char (specific surface area 2.0 m² g⁻¹) and of activated carbon (specific surface area approximately 900 m² g⁻¹) on an extracellular enzymatic reaction involved in the degradation of cellulose (i.e., hydrolysis of cellobiose by β-glucosidase from Aspergillus niger) were investigated. Cellobiose was not adsorbed by chestnut wood char, whereas activated carbon absorbed more than 97% of it. Both charred materials adsorbed more than 99% of β-glucosidase. For chestnut wood char, adsorption of the enzyme caused a decrease of approximately 30% in the reaction rate, whereas for activated carbon, the nearly complete absorption of both substrate and enzyme entirely inhibited the reaction. These results show that β-glucosidase from A. niger retains most of its activity when adsorbed to chestnut wood char and that the reaction it catalyses in nature is only slightly affected by this charred material. On the other hand, a material characterised by a high specific surface area and high porosity, such as activated carbon, can make even a highly soluble substrate unavailable for soil enzymes and therefore completely inhibit the reaction. Thus, charred materials may affect nutrient cycling mainly by regulating the availability of substrates: the degradation of highly soluble substrates may be accelerated by materials with low specific surface area, which maintain an active and protected enzyme pool, whereas materials with high specific surface and high porosity may slow down the degradation by making substrates unavailable.     

Greenhouse gas diffusive fluxes at the sediment–water interface of sewage-draining rivers

Purpose

The purposes of this study were to analyse the spatiotemporal variations in greenhouse gas diffusive fluxes at the sediment–water interface of sewage-draining rivers and natural rivers, and investigate the factors responsible for the changes in greenhouse gas diffusive fluxes.

Materials and methods

Greenhouse gas diffusive fluxes at the sediment–water interface of rivers in Tianjin city (Haihe watershed) were investigated during July and October 2014, and January and April 2015 by laboratory incubation experiments. The influence of environmental variables on greenhouse gas diffusive fluxes was evaluated by Spearman’s correlation analysis and a multiple stepwise regression analysis.

Results and discussion

Sewage-draining rivers were more seriously polluted by human sewage discharge than natural rivers. The greenhouse gas diffusive fluxes at the sediment–water interface exhibited obvious spatiotemporal variations. The mean absolute value of the CO₂ diffusive fluxes was seasonally variable with spring>winter>fall>summer, while the mean absolute values of the CH₄ and N₂O diffusive fluxes were both higher in summer and winter, and lower in fall and spring. The annual mean values of the CO₂, CH₄ and N₂O diffusive fluxes at the sewage-draining river sediment–water interface were ??123.26?±?233.78 μmol m^?2 h^?1, 1.88?±?6.89 μmol m^?2 h^?1 and 1505.03?±?2388.46 nmol m^?2 h^?1, respectively, which were 1.22, 4.37 and 134.50 times those at the natural river sediment–water interface, respectively. The spatial variation of the N₂O diffusive fluxes in the sewage-draining rivers and the natural rivers was the most significant. As a general rule, the more serious the river pollution was, the greater the diffusive fluxes of the greenhouse gases were. On average for the whole year, the river sediment was the sink of CO₂ and the source of CH₄ and N₂O. There were positive correlations among the CO₂, CH₄ and N₂O diffusive fluxes. The main influencing factor for CO₂ and N₂O diffusive fluxes was the water temperature of the overlying water; however, the key factors for CH₄ diffusive fluxes were the Eh of the sediment and the NH₄⁺-N of the overlying water.

Conclusions

River sediment can be either a sink or a source of greenhouse gases, which varies in different levels of pollution and different seasons. Human sewage discharge has greatly affected the carbon and nitrogen cycling of urban rivers.

     

Glucose slows down the heat-induced aggregation of β-lactoglobulin at neutral pH

The behavior of β-lactoglobulin (β-Lg) during heat treatments depends on the environmental conditions. The influence of the presence or absence of a reducing sugar, namely, glucose, on the modification of the protein during heating has been studied using fluorescence, polyacrylamide gel electrophoresis (PAGE), size-exclusion chromatography (SEC), and transmission electron microscopy. Glycated products were formed during heating 24 h at 90 °C and pH 7. The fluorescence results revealed an accumulation of the advanced Maillard products and the formation of aggregates during heating. PAGE and SEC data suggested that the products in the control samples were essentially composed of covalently linked fibrillar aggregates and that their formation was faster than that for glycated samples. We showed that glucose affected the growing step of covalent aggregates but not the initial denaturation/aggregation step of native protein. Glucose-modified proteins formed a mixture of short fibrils and polydisperse aggregates. Our results revealed that β-Lg forms fibrils at neutral pH after heating and that glucose slows the formation of these fibrils.     

Microbial populations and the activity of the soil under agricultural and agricultural–pastoral systems

The effects of agricultural–pastoral and tillage practices on soil microbial populations and activities have not been systematically investigated. The effect of no-tillage (NT), no-tillage agricultural–pastoral integrated systems (NT-I) and conventional tillage (CT) at soil depths of 0–10, 10–20 and 20–30 cm on the microbial populations (bacteria and fungi), biomass-C, potential nitrification, urease and protease activities, total organic matter and total N contents were investigated. The crops used were soybean (in NT, NT-I and CT systems), corn (in NT and NT-I systems) and Tanner grass (Brachiaria sp.) (in NT-I system); a forest system was used as a control. Urease and protease activities, biomass-C and the content of organic matter and total N were higher (p < 0.05) in the forest soil than the other soils. Potential nitrification was significantly higher in the NT-I system in comparison with the other systems. Bacteria numbers were similar in all systems. Fungi counts were similar in the CT and forest, but both were higher than in NT. All of these variables were dependent on the organic matter content and decreased (p < 0.05) from the upper soil layer to the deeper soil layers. These results indicate that the no-tillage agricultural–pasture-integrated systems may be useful for soil conservation.     

Isolation and identification of sea buckthorn (Hippophae rhamnoides) phenolics with antioxidant activity and α-glucosidase inhibitory effect

This study was performed to evaluate the antioxidant and α-glucosidase inhibitory effects from the extract, fractions, and isolated compounds of sea buckthorn leaves. Six compounds, kaempferol-3-O-β-D-(6'-O-coumaryl) glycoside, 1-feruloyl-β-D-glucopyranoside, isorhamnetin-3-O-glucoside, quercetin 3-O-β-D-glucopyranoside, quercetin 3-O-β-D-glucopyranosyl-7-O-α-L-rhamnopyranoside, and isorhamnetin-3-O-rutinoside, were isolated from sea buckthorn leaf extracts. The butanol fraction (EC(50) = 1.81 μg/mL) along with quercetin 3-O-β-D-glucopyranoside (EC(50) = 1.86 μg/mL) had a higher DPPH radical-scavenging activity and showed stronger reducing power (OD(700) = 1.83 and 1.78, respectively). The butanol fraction (477 mg GAE/g) contained the highest amount of phenolic compounds and also the most powerful α-glucosidase inhibitory effect (86%) at 5 μg/mL. The results indicate that sea buckthorn leaf extracts could potentially be used for food additives and the development of useful natural compounds.