Tannic acid reduces recovery of water-soluble carbon and nitrogen from soil and affects the composition of Bradford-reactive soil protein

Tannins are plant-derived polyphenolic compounds that precipitate proteins, bind to metals and complex with other compounds. Solutions of tannic acid, or other phenolic compounds, were added to soil samples to determine if they would affect recovery of soluble soil carbon (WSC) or –nitrogen (WSN) or influence the extraction and composition of Bradford-reactive soil protein (BRSP), associated with glomalin. Tannic acid-C added with water was not completely recovered from samples and the amount of total net WSC and WSN recovered was reduced, suggesting formation of insoluble complexes. By comparison, non-tannin phenolics like gallic acid, or methyl gallate, had little effect on extraction of WSC or WSN while a simple gallotannin derived from tannic acid, 1,2,3,4,6-penta-O-galloyl-d-glucose (PGG), inhibited extraction most. The C and N concentrations in BRSP increased when soil samples were treated with tannic acid or PGG before extraction, a procedure that includes autoclaving. Increases were greatest in the 10–20 cm compared to 0–5 cm depth. Accompanying these were declines in the ratio of absorbance at 465 and 665 nm (E4/E6 ratio) of BRSP extracts suggesting formation of larger or heavier molecules. In contrast, C and N composition in lyophilized BRSP was unaffected or even slightly reduced when tannic acid or PGG were added to the BRSP extract solution after the extraction process. We conclude that some tannins can reduce the solubility of labile soil C and N, at least temporarily and given unpredictability of response associated with phenolic substances, the Bradford assay should not be relied on to quantify pools or composition of soil proteins like glomalin.     

Behavior of Bradford-reactive substances is consistent with predictions for glomalin

There is considerable controversy concerning detection in soils of the protein, glomalin, which is produced by arbuscular mycorrhizal fungi. Glomalin was originally defined as a substance that cross reacts with a monoclonal antibody formed against a substance in the cell walls of an arbuscular mycorrhizal fungus. Thus, one can use an immunological approach to detect glomalin. However, it was recently shown that other proteins cross react with the antibody. The other, more common, approach involves assay of soil protein using the Bradford reaction. This approach assumes that the Bradford assay is specific to protein, and that the assayed protein is largely glomalin, either because other proteins are in low concentration, or because the extraction process eliminates the possibility of their detection. These assumptions, however, have been called into question recently. One way to test whether the Bradford assay can be useful in quantifying glomalin is to determine whether the concentrations of Bradford-reactive substances are consistent with predictions for glomalin. For example, if recently produced glomalin is more labile than older glomalin, the concentrations of the two fractions should not be highly correlated. Moreover, when a contrast is established between mycorrhizal and nonmycorrhizal vegetation, recently produced glomalin should soon occur in higher concentration in soils supporting mycorrhizal vegetation. Older glomalin should not be found in higher concentrations in the soils of mycorrhizal vegetation until some time later. We tested these predictions by employing the Bradford assay during the course of a three-year field experiment in which canola (nonmycorrhizal) and soy (mycorrhizal) were grown in separate plots in year 1, both of which were followed by maize (mycorrhizal) in years 2 and 3. The correlation between the concentrations of fraction 1 Bradford-reactive substances (also known as easily extractable glomalin and frequently assumed to be recently produced) and fraction 2 (the more difficult-to-extract fraction and frequently assumed to be older glomalin), was very poor. In year 1, the concentration of fraction 1 was significantly greater in soy plots than in canola plots. Finally, fraction 2 was only significantly higher in the former soy plots than in former canola plots in years 2 and 3. These data support the hypothesis that the Bradford assay was useful in detecting glomalin in this case.     

Glomalin‐related soil protein in French temperate forest soils: interference in the Bradford assay caused by co‐extracted humic substances

Thermostable soil protein, known as glomalin, is an important component of soil carbon stocks. Thought to originate from endomycorrhizal fungi, Glomales, this operationally‐defined fraction of soil organic matter contains proteins of diverse origin as well as non‐protein material, including humic substances. Accumulation results from the balance between production/release and subsequent degradation. Quantification of the protein is subject to uncertainty because of the co‐extraction of other components that interfere with the Bradford assay. We studied 10 topsoils from French temperate forests, taken from the national forest monitoring network (Renecofor). Two fractions were extracted, easily extractable (EE) at neutral pH and total extractable (T) at pH 8. Protein was quantified with the colorimetric Bradford method, either by direct calibration using bovine serum albumin (BSA) or by extrapolation of the standard addition plot of BSA. Solubilized organic matter was characterized by using absorbance at 465 and 665 nm and by three‐dimensional fluorescence excitation‐emission spectroscopy. Neither soil properties nor forest cover influenced glomalin‐related soil protein (GRSP) content. Direct assay gave the GRSP_EE to be about 1 g kg^?1 soil, and GRSP_T in the range 3–10 g kg^?1, accounting for about 2% of soil organic carbon and about 15% of soil nitrogen. Standard addition plots indicated a two to sixfold under‐estimation of protein in total extracts, caused by negative interference with the Bradford assay. The GRSP_EE was correlated significantly with both estimates of GRSP_T. Under‐estimation of GRSP_T by direct assay was not related to the E4:E6 ratio but was correlated significantly with the intensity of absorbance at either 460 or 660 nm and with one of the fluorescence peaks. We conclude that GRSP_EE is not necessarily more recent than GRSP_T and that both fractions may be probes of protein content, but that absolute contents may be under‐estimated because of co‐extracted humic substances.     

Glomalin-related soil protein: Assessment of current detection and quantification tools

Despite the widely acknowledged importance of arbuscular mycorrhizal fungi (AMF) in soil ecology, quantifying their biomass and presence in field soils is hindered by tedious techniques. Hence biochemical markers may be useful, among which glomalin-related soil protein (GRSP) could show a particular promise. Presently GRSP is operationally defined, its identification resting solely on the methods used to extract it from soil (citric acid buffer and autoclaving) and the assays (Bradford/enzyme-linked immunosorbent assay (ELISA) with a monoclonal antibody) utilized to detect it. The current assumption is that most non-heat stable soil proteins except glomalin are destroyed during the harsh extraction procedure. However, this critical assumption has not been tested. The purpose of this research was to challenge the GRSP extraction process to determine the accuracy of the Bradford method as a measure of glomalin; and to provide some assessment of the specificity of the ELISA monoclonal antibody. In two studies we spiked soil samples either with known quantities of a glycoprotein (BSA: bovine serum albumin) or with leaf litter from specific sources. After extraction 41-84% of the added BSA was detected with the Bradford method. This suggests that the currently used extraction procedure does not eliminate all non-glomalin proteins. Also, ELISA cross-reactivity against BSA was limited, ranging from 3% to 14%. Additions of leaf litter also significantly influenced GRSP extraction and quantification suggesting that plant-derived proteins, as would occur in the field, had a similar effect as BSA. Litter additions decreased the immunoreactive protein values, suggesting interference with antibody recognition. We conclude that the use of GRSP, especially Bradford-based detection, in the assessment of AMF-derived substances within field soils is problematic, it may be inappropriate in situations of significant organic matter additions.     

Bradford-reactive soil proteins and aggregate stability under abandoned versus tilled olive groves in a semi-arid calcisol

Glomalin, a substance produced by arbuscular mycorrhizal fungi, is reported to play a role in soil aggregation, but this role has been questioned in soils rich in calcium carbonate. We studied the relationship between aggregation stability and glomalin in a Haplic Calcisol comparing abandoned and active cultivation of olive groves. Abandonment was associated with increases in soil organic carbon, the percentage of water stable aggregates (WSA_1-2mm), and easily extractable and total Bradford-reactive soil protein. WSA_1-2mm was strongly positively correlated with both easily extractable and total Bradford-reactive soil protein. While easily extractable Bradford-reactive soil protein measured in both stable and unstable aggregates did not show any significant differences, Bradford-reactive soil protein was twice as high in stable than in unstable aggregates under both tillage and abandonment. Our results suggest that Bradford-reactive soil protein influences aggregate stability, even in soils with low organic matter and high calcium carbonate contents. However, more research is needed to elucidate the role of easily extractable Bradford-reactive soil protein in soil aggregation.     

Polyphenolic compounds interfere with quantification of protein in soil extracts using the Bradford method

 The Bradford protein quantification assay is based on an absorbance shift in Coomassie brilliant blue G-250 (CBB). Samples extracted for glomalin, a protein produced by arbuscular mycorrhizal (AM) fungi, are quantified using the Bradford assay. CBB is known to react with polyphenolic substances, and co-extraction of glomalin and humic substances is known to occur. The effects of increasing concentrations polyphenolic compounds were measured. The addition of any amount of polyphenolic compounds increased the Bradford reactive fraction (BRF) of soil extract. Caution is required when interpreting BRF data, as comparison of BRF data from different studies or different field sites is problematic. The BRF may represent recalcitrant organic material in soil, though its relationship to AM fungi remains unclear.     

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.     

Glomalin-related soil protein contains non-mycorrhizal-related heat-stable proteins, lipids and humic materials

Glomalin is reportedly a stable and persistent protein produced in copious quantities by mycorrhizal fungi and may be an important pool of organic N in soil. Glomalin-related soil protein (GRSP), however, is only operationally defined by its extraction method, and has been only poorly characterized at best. The goal of this study was to characterize the molecular structures within GRSP. Synchrotron-based X-ray absorption near-edge structure (XANES) spectroscopy and pyrolysis field-ionization mass spectrometry (Py-FIMS) revealed that GRSP contains a consortium of proteins along with many impurities. Employing proteomic techniques, we found that glomalin itself may be a thioredoxin-containing chaperone; however, no homologies with proteins or DNA of mycorrhizal origin were detected. Proteomics techniques further revealed that this fraction contains large amounts of soil-related heat-stable proteins and proteins of non-mycorrhizal origin. Results of this research show that the current extraction procedure that defines GRSP yields a mixture of compounds and thereby overestimates glomalin stocks when quantified using the Bradford assay. The chemical nature of glomalin has yet to be conclusively determined; it is unlikely that the chemical structure of glomalin can be elucidated from the mixture extracted as GRSP. Instead, an investigation into the specific biochemistry of immunoreactive assays currently used to define GRSP, followed by proteomic characterization of monoxenic mycorrhizal cultures may be required to advance our understanding of the chemical nature and agronomic significance of GRSP in soils.     

Carbon and nitrogen in operationally defined soil organic matter pools

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

Applying an oxygen-based respiratory assay to assess soil microbial responses to substrate and N availability

Documented approaches for measuring soil microbial activities and their controlling factors under field conditions are needed to advance understanding of soil microbial processes for numerous applications. We manipulated field plots with carbon (C) and nitrogen (N) additions to test the capability of a respiratory assay to: (1) measure respiration of endogenous soil C in comparison to field-measured CO₂ fluxes; (2) determine substrate-induced respiratory (SIR) activities that are consistent with substrate availability in the field; and, (3) report N availability in the field based on assay responses with and without added N. The respiratory assay utilizes a microplate containing an oxygen-sensitive fluorescent ruthenium dye. Respiratory activities measured with this approach have previously been shown to occur within short (6–8 h) incubation periods using low substrate concentrations that minimize enrichment during the assay. Field treatments were conducted in a randomized full-factorial design with C substrate (casamino acids, glucose, or none) and inorganic N (±) as the treatment factors. With one exception, we found that respiration of endogenous soil C in the assay responded to the field treatments in a similar manner to CO₂ fluxes measured in the field. Patterns of SIR with low concentrations of added amino acid or carbohydrate substrate (200 μg C g⁻¹ soil) were consistent with field treatments. The ratio (N_ratio) of carbohydrate respiration with added N (25 μg N g⁻¹ soil) to the same without N in the assay was significantly (P < 0.05) decreased by field N amendment. The carbohydrate N_ratio exhibited a logarithmic relationship (r = 0.64, P < 0.05) with extractable inorganic soil nitrate and ammonium concentrations. These data significantly extend and support the capability of this oxygen-based respiratory assay to evaluate in situ soil activities and examine factors that limit these activities.     

Role of allophanes in the accumulation of glomalin‐related soil protein in tropical soils (Martinique,French West Indies)

Thermo‐stable, operationally defined soil protein, known as glomalin, may make an important contribution to carbon storage in soils. The term glomalin is used because this putative protein, or group of proteins, was originally thought to be produced only by Glomus fungi. There is currently little information on the glomalin‐related soil protein (GRSP) content of tropical soils, particularly allophanic soils that are known to have different carbon dynamics to temperate climate soils. We have measured the Bradford‐reactive GRSP content of soils sampled from forests and grasslands on the tropical island of Martinique and compared the observations with soil composition. Two operationally defined fractions of GRSP were measured, namely easily‐extractable and total GRSP. The contents of GRSP in moist soils were in the range of 2–36 g kg^?1, accounting for about 8% of soil organic carbon, and were greater in topsoils than in corresponding subsoils. Both the GRSP contents and the fraction of soil organic carbon attributed to GRSP were greater than those reported for temperate climate soils. Both total and easily extractable GRSP contents were positively correlated to soil organic carbon content. The fraction of soil organic carbon that could be attributed to soil protein decreased with increasing allophane content for allophanic soils. No other trends of GRSP content with soil properties or land use were found. GRSP extraction was decreased about seven‐fold by air‐drying of soils, confirming the irreversible change in the soil microstructure of allophanic soils. Total and easily extractable GRSP were correlated and we conclude that both are good probes of thermo‐stable soil protein content for these soils. No attempt was made to verify the fungal origin of the protein detected.     

Stabilization of oxidative enzymes in desert soil may limit organic matter accumulation

Phenol oxidase and peroxidase activities in desert grassland soils at the Sevilleta Long Term Ecological Research site in central New Mexico (USA) are far greater than those of temperate soils. Activity is uniformly distributed across particles ranging from >1 mm to <38 μm and is unaffected by autoclaving, in contrast to hydrolase activities. The sorbed enzymes are readily extractable and inactivated by boiling. High soil pH, high stabilized oxidative enzyme activity, and carbonates create optimal conditions for degradation of phenols which increase decomposition potentials and limit soil organic matter accumulation.     

A quick and sensitive method for the quantification of peroxidase activity of organic surface soil from forests

The purpose of the present study was to test the non-mutagenic compound 3,3′,5,5′-tetramethylbenzidine (TMB) as a model substrate for peroxidase in forest topsoil, as an alternative to the conventional substrate l-3,4-dihydroxyphenylalanine (l-DOPA). TMB was highly sensitive; linear absorbance changes of 0.6 were achieved within 20 min for 1000-fold diluted soil. Brief heating (denaturation) of the soil suspension gave a 34-fold reduction of TMB oxidation, indicating that the reaction measured by TMB was indeed an enzymatic reaction. TMB oxidation showed a narrow peak at pH 4.4. A proportional decrease in peroxidase activity, when the soil suspension was diluted, demonstrated that TMB estimates of peroxidase activity are directly comparable when corrected for differences in sample size. Oxidation of TMB was slow in the absence of H₂O₂ suggesting that TMB is a poor substrate for phenol oxidases. TMB oxidation was tested in nine different forest topsoils. The peroxidase activity, when normalised to the amount of soil organic matter, ranged from 1.4±0.1 Δabs₄₅₀ h^?1 mg^?1 to 34.9±4.3 Δabs₄₅₀ h^?1 mg^?1. In comparison, l-DOPA oxidation by soil peroxidases and commercial peroxidases gave inconsistent results, suggesting that one should be cautious when using l-DOPA as a soil peroxidase substrate. The high sensitivity of TMB, compared to l-DOPA, and the low interference from phenol oxidase and humic substances suggest that TMB is a better substrate than l-DOPA for estimation of peroxidase activity of forest topsoil.     

Improved soil structure and citrus growth after inoculation with three arbuscular mycorrhizal fungi under drought stress

In a controlled potted experiment, citrus (Poncirus trifoliata) seedlings were inoculated with three species of arbuscular mycorrhizal (AM) fungi, Glomus mosseae, G. versiforme or G. diaphanum. Two soil-water levels (ample water, −0.10 MPa; drought stress, −0.44 MPa) were applied to the pots 4 months after transplantation. Eighty days after water treatments, the soils and the citrus seedlings were well colonized by the three AM fungi. Mycorrhizal fungus inoculation improved plant biomass regardless of soil-water status but decreased the concentrations of hot water-extractable and hydrolyzable carbohydrates of soils. Mycorrhizal soils exhibited higher Bradford-reactive soil protein concentrations than non-mycorrhizal soils. Mycorrhizas enhanced >2 mm, 1–2 mm and >0.25 mm water-stable aggregate fractions but reduced 0.25–0.5 mm water-stable aggregates. Peroxidase activity was higher in AM than in non-AM soils whether drought stressed or not, whereas catalase activity was lower in AM than non-AM soils. Drought stress and AM fungus inoculation did not affect polyphenol oxidase activity of soils. A positive correlation between the Bradford-reactive soil protein concentrations, soil hyphal length densities, and water-stable aggregates (only >2 mm, 1–2 mm and >0.25 mm) suggests beneficial effects of the AM symbiosis on soil structure. It concluded that AM fungus colonization enhanced plant growth under drought stress indirectly through affecting the soil moisture retention via glomalin's effect on soil water-stable aggregates, although direct mineral nutritional effects could not be excluded.     

Detection and quantification of the nematophagous fungus Hirsutella minnesotensis in soil with real-time PCR

A real-time PCR assay was developed to quantify in soil the fungus Hirsutella minnesotensis, an important parasite of secondary-stage juvenile (J2) of the soybean cyst nematode. A primer pair 5′-GGGAGGCCCGGTGGA-3′ and 5′-TGATCCGAGGTCAACTTCTGAA-3′ and a TaqMan probe 5′-CGTCCGCCGTAAAACGCCCAAC-3′ were designed based on the sequence of the ITS region of the rRNA gene. The primers were highly species-specific. The PCR reaction system was very sensitive and able to detect as few as 4 conidia g^?1 soil. Regression analysis showed similar slopes and efficiency on DNA from pure culture (y = ?3.587x + 41.017, R² = 0.9971, E = 0.9055) and from Log conidia g^?1 soil (y = ?3.855x + 37.669, R² = 0.9139, E = 0.8172), indicating that the real-time PCR protocol can reliably quantify H. minnesotensis in the soil. The real-time PCR assay was applied to 20 soil samples from soybean fields, and compared with a parasitism assay. The real-time PCR assay detected H. minnesotensis in six of the soils, whereas the parasitism assay detected H. minnesotensis in the same six soils and three additional soils. The real-time PCR assay was weakly correlated (R² = 0.49) with the percentage of parasitized J2 in the six soils, indicating that different types of soil may interfere the efficiency of the real-time PCR assay, possibly due to the effect of soil types on efficacy of DNA extraction. The parasitism assay appeared to be more sensitive than real-time PCR in detecting presence of H. minnesotensis, but real-time PCR was much faster and less costly and provided a direct assessment of fungal biomass. Using the two assays in combination can obtain more complete information about the fungus in soil than either assay alone. Hirsutella parasitism was widespread and detected in 13 of the 20 field soils, indicating that these fungi may contribute to suppressiveness of soybean cyst nematode in nature and likely have high biological control potential for the nematode.     

Relationships of soil properties with Mn and Zn distribution in acidic soils and their uptake by a barley crop

The extractability and solid-phase fractionation of manganese (Mn) and zinc (Zn) in acid-to-neutral agricultural soils from Central Spain was evaluated by sampling and analysing twenty-nine representative soils and by greenhouse cropping eleven of them with spring barley (Hordeum vulgare, L.). All soil samples were extracted with three chemical extractants commonly used for soil fertility evaluation (0.43 M HOAc, DTPA and Mehlich-3). The soil samples were also operationally determined in six steps with the following extractants: 1 M Mg(NO₃)₂ extractable (WSEX, water soluble plus exchangeable), 0.7 M NaOCl extractable (OC, organically complexed), 0.1 M NH₂OH·HCl extractable (MnOX, Mn-oxide), 0.2 M (NH₄)₂C₂O₄ + 0.2 M H₂C₂O₄ extractable (AFeOX, amorphous Fe-oxide), 0.2 M (NH₄)₂C₂O₄ + 0.2 M H₂C₂O₄ + 0.1 M ascorbic acid extractable (CFeOX, crystalline Fe-oxide), and HCl, HNO₃, and HF in mixture (RES, residual). Soil-extractable amounts for the three single extractants were highly correlated with each other for both metals. Distributions among metal fractions showed that Mn was mainly found in the MnOX fraction (30.9%, ranging from 13.0 to 51.2%), whereas Zn was predominantly found in the RES fraction (44.3%, ranging from 26.4 to 56.8%). The proportion of Mn fractions extracted from the soils was in the order as follows: CFeOX ≤ WSEX = OC ≤ AFeOX = RES < MnOX, whereas Zn was in the order: WSEX ≤ OC ≤ AFeOX < MnOX < CFeOX < RES. The soil properties that correlated best with the distribution of Mn and Zn forms in these soils were soil organic matter and pH. The “availability factor” values [AF = (WSEX + OC) 100 / total metal], were higher for Mn than for Zn in these soils. Plant metal concentrations (Y) and soil-extractable and sequential extracted fractions showed few significant correlations. However, it was possible to significantly predict the phytoavailability of Mn and Zn for barley using a series of empirical equations involving extractable metals, solid-phase fractions and soil properties as components. The R² values of the best-fit regression models ranging from 0.50 [Y-Zn = 19.3 + 6.32 (WSEX + OC)-Zn] to 0.92 [Y-Zn = 57.3 + 0.23 P − 8.56 pH + 20.6 DTPA-Zn].     

Effects of tillage and nitrogen management on soil chemical and physical properties after 23 years of continuous sorghum

Long-term tillage and nitrogen (N) management practices can have a profound impact on soil properties and nutrient availability. A great deal of research evaluating tillage and N applications on soil chemical properties has been conducted with continuous corn (Zea Mays L.) throughout the Midwest, but not on continuous grain sorghum (Sorghum bicolor (L.) Moench). The objective of this experiment was to examine the long-term effects of tillage and nitrogen applications on soil physical and chemical properties at different depths after 23 years of continuous sorghum under no-till (NT) and conventional till (CT) (fall chisel-field cultivation prior to planting) systems. Ammonium nitrate (AN), urea, and a slow release form of urea were surface broadcast at rates of 34, 67, and 135 kg N ha⁻¹. Soil samples were taken to a depth of 15 cm and separated into 2.5 cm increments. As a result of lime applied to the soil surface, soil pH in the NT and CT plots decreased with depth, ranging from 6.9 to 5.7 in the NT plots and from 6.5 to 5.9 in the CT plots. Bray-1 extractable P and NH₄OAc extractable K was 20 and 49 mg kg⁻¹ higher, respectively, in the surface 2.5 cm of NT compared to CT. Extractable Ca was not greatly influenced by tillage but extractable Mg was higher for CT compared to NT below 2.5 cm. Organic carbon (OC) under NT was significantly higher in the surface 7.5 cm of soil compared to CT. Averaged across N rates, NT had 2.7 Mg ha⁻¹ more C than CT in the surface 7.5 cm of soil. Bulk density (Δ_b) of the CT was lower at 1.07 g cm⁻³ while Δ_b of NT plots was 1.13 g cm⁻³. This study demonstrated the effect tillage has on the distribution and concentration of certain chemical soil properties.     

Microbial biomass C measurements in soil of the central highlands of Mexico

Microbial biomass is the key factor in nutrient dynamics in soil, but no information exist about it in soils of the central highlands of Mexico, a major agricultural area. We determined the microbial biomass in soils with a wide range of organic C and belonging to three soil texture classes. Twenty-four soils under different types of cultivation were sampled while microbial biomass C was measured with the chloroform fumigation incubation (CFI) and extraction technique (CFE). Microbial biomass C as measured with the CFI technique ranged from 138 to 2195 mg C kg⁻¹. The ninhydrin-positive compounds (NPC) and extractable C released with CFE increased with increased time of exposure to chloroform and on average 53% of NPC and 83% of extractable C was released after 1 day compared to that released after 10 days. The ratio of microbial biomass C as measured with the CFI method related to the NPC was 31.8 after 1 day and 20.0 after 10 days while the relationship with extractable C was 3.18 and 2.69, respectively. The relationship between microbial biomass C as measured by the chloroform fumigation incubation technique and the soluble C and ninhydrin-N rendered extractable after 1 and 10 days of chloroform fumigation for soils of the central highlands of Mexico were comparable to values reported for soils in other regions of the world. The factors determined in this study can thus be used to determine microbial biomass.     

Enzyme activities and diuron persistence in soil amended with vermicompost derived from spent grape marc and treated with urea

Mineral fertilizers, organic amendments, and pesticides are inputs commonly used in conventional farming practices. The aim of this study was to evaluate the effects of single or combined applications of spent grape marc-vermicompost, urea, and/or diuron on soil-enzyme activities and the persistence of this herbicide in soils with low organic carbon content. The application of vermicompost enhanced dehydrogenase (DHase) enzyme activity over time but altered soil urease activity to a very limited extent. The reduction in diuron concentrations and the increase in DHase activity indicated that the soil microorganisms were capable of degrading the ureic herbicide. Treatment with vermicompost and diuron had a stimulatory effect on soil microbial activity. On the whole, the application of diuron and urea to the vermicompost-amended soil raised DHase and urease activity to maximum levels (>3 μg INTF g^?1 h^?1 and >47 μg NH₄⁺ g^?1 h^?1, respectively). The application of urea to the unamended and vermicompost-amended soil decreased diuron persistence from 18.8 and 33 d to 12.5 and 15 d, respectively. Our findings show that although vermicompost additions reduce diuron availability, this boosts diuron degradation when combined with urea. These additions, under different soil management conditions, minimize the bioavailability and persistence of diuron and consequently the risk of leaching and seepage into aquifers. Compared with untreated soils, these types of treated soils could also improve agricultural sustainability and the quality of the environment.     

Microbial biomass in a semi arid soil of the central highlands of Mexico cultivated with maize or under natural vegetation

Microbial biomass (MB) is the key factor in nutrient dynamics in soil, but no information exists how clearing of vegetation to cultivate maize in the central highlands of Mexico might affect it. Soil MB was measured with the chloroform fumigation incubation (CFI) and fumigation extraction (CFE) techniques and the substrate-induced respiration (SIR) method in soil sampled under or outside the canopy of mesquite (Prosopis laevigata) and huisache (Acacia tortuoso), N₂ fixing shrubs, and from fields cultivated with maize. Microbial biomass C as measured with the CFI technique ranged from 122 mg C kg⁻¹ in agricultural soil to 373 mg C kg⁻¹ in soil sampled under mesquite shrubs. Microbial biomass N as measured with the CFI technique ranged from 11 mg N kg⁻¹ in agricultural soil to 116 mg N kg⁻¹ in soil sampled under mesquite shrub. The ratio of microbial biomass C as measured with CFI related to the ninhydrin-positive compounds (NPC) was 12.23 after 1 day and 8.43 after 10 days while the relationship with extractable C was 3.15 and 2.96, respectively. The metabolic quotient (qCO₂) decreased in the order OUTSIDE > MESQUITE > HUIZACHE > AGRICULTURE, and the microbial biomass:soil organic C ratio decreased in the order MESQUITE > HUIZACHE > OUTSIDE > AGRICULTURE using SIR to determine the microbial biomass. It was found that converting soil under natural vegetation to arable soil was not only detrimental for soil quality, but might be unsustainable as organic matter input is limited.