Repeated applications of tannins and related phenolic compounds are retained by soil and affect cation exchange capacity

Retention of tannins, produced by plants, could be important for managing soil organic matter and nutrient cycling. However, we know little about the comparative retention of different classes of tannins and related compounds or if soils have a maximum storage capacity for them. To address these questions, forest, and pasture loam soils, collected at 0-5 cm (surface) and 10-20 cm (subsurface), were repeatedly treated with water (Control) or solutions containing condensed and hydrolyzable tannins or related phenolic subunits (10 mg g⁻¹ soil). Treatments included a polymeric flavonoid-based procyanidin from sorghum, catechin, tannic acid, β-1,2,3,4,6-penta-O-galloyl-d-glucose (PGG), gallic acid, and methyl gallate. After each application, soluble-C in supernatants was determined by oxidative-combustion infrared analysis and retention of treatment-carbon by soil was calculated as the difference between added and recovered soluble-C. An interaction between soil depth and treatment was evident through all applications with highest retention of both hydrophobic (PGG) and hydrophilic (procyanidin) tannins, compared to other phenolic compounds. For all treatments except gallic acid and methyl gallate, higher sorption occurred in surface soil, which contained more organic matter than subsurface soil. With each successive application, less additional treatment-C was retained by soil and the amount of C remaining in supernatants was correlated with the presence of phenolic substances. Cumulative retention by surface soil was more than 10.3, 8.5 and 6.4 mg C g⁻¹ soil for PGG, tannic acid, and procyanidin, several times higher than the other compounds. Soluble-C extracted from treated soil, with cool water (23 °C), was 1-2 orders of magnitude greater than Control samples and highly correlated with Prussian Blue (PB) phenolics, indicating some retained treatment-C was only weakly held on the soil. The final extraction, with hot water (80 °C), removed more soluble-C, particularly from surface samples, that contained fewer PB phenolics per unit soluble-C than cool water extracts. After all extractions more than 85% of sorbed procyanidin-C was retained by samples compared to 81% of methyl gallate, 79% of PGG, 74% of tannic acid, 50% of catechin, and 40% of the gallic acid. Total C, measured in soil after all extractions, was close to expected values, confirming tannins and phenolic compounds had remained in soil and were not otherwise lost. Cation exchange capacity was increased about 30% in subsurface and forest samples by PGG, a hydrolyzable tannin, but decreased by 30% and 35% in surface and pasture soil, respectively, by its monomer, gallic acid.     

Glomalin extraction and measurement

We investigated extraction from soil of glomalin, a glycoprotein produced by arbuscular mycorrhizal fungi, and we examined its measurement. The most commonly used protocols for extracting glomalin require autoclaving of soil in citrate solution, followed by centrifugation to separate the supernatant, and then measurement by either Bradford protein assay or enzyme-linked immunosorbent assay (ELISA). We found that lengthening the time of autoclaving increased easily extractable glomalin extraction. Delay of centrifugation after autoclaving, however, diminished Bradford-reactive substances in the supernatant, suggesting that extracted substances might be reversibly immobilized on soil particles. Surprisingly, increasing the volume of extraction solution did not accelerate extraction of “total glomalin”, but instead, substantially increased the amount extracted. Multiple autoclave cycles nevertheless denature glomalin, which may not be as heat-resistant as thought. Repeated 1-h autoclaving of supernatant diminished both its Bradford-reactive substances (7.3% h^?1) and immunoreactive protein (22% during the first hour and 9.5% h^?1 of the remainder thereafter), although a large initial volume of extractant could reduce the loss of immunoreactive protein. Proteins and polyphenols that survive the extraction process are measured non-specifically by the Bradford assay. When we added other glycoproteins to dry soils, we recovered a maximum 34% bovine serum albumin and 22% bovine mucin, primarily in the first two, 1-h extraction cycles. These added proteins may adhere to soil organic matter and thereby be protected from denaturation. In addressing the endpoint of glomalin extraction, we found that the Michaelis–Menten equation closely fits cumulative glomalin per extraction cycle such that its asymptote provides an objective estimate of total extractable glomalin for a given set of extraction conditions. Additionally, the equation provides a curvature parameter that reflects the soil-specific efficiency of an extraction protocol. Although the soils that we investigated with 7.6% or more soil organic matter had the most asymptotic total glomalin, they were extracted the least efficiently.     

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.     

Spatial distribution of arbuscular mycorrhizal fungi, glomalin and soil enzymes under the canopy of Astragalus adsurgens Pall. in the Mu Us sandland, China

To measure and manage plant growth in arid and semi-arid sandlands, improved understanding of the spatial patterns of desert soil resources and the role of arbuscular mycorrhizal (AM) fungi is needed. Spatial patterns of AM fungi, glomalin and soil enzyme activities were investigated in five plots located in the Mu Us sandland, northwestern China. Soils to 50 cm depth in the rhizosphere of Astragalus adsurgens Pall. were sampled. The study demonstrated that A. adsurgens Pall. could form strong symbiotic relationships with AM fungi. Arbuscular mycorrhizal fungal status and distributions were significantly different among the five studied plots. Correlation coefficient analysis demonstrated that spore density was significantly and positively correlated with soil organic carbon (SOC), soil acid phosphatase and to two Bradford-reactive soil protein (BRSP) fractions (P < 0.01). Colonization of arbuscules and vesicles were positively correlated with protease activity. The BRSP fractions were also significantly and positively correlated to edaphic factors (e.g. SOC, available nitrogen, and Olsen phosphorus) and soil enzymes (e.g. soil urease and acid phosphatase). The means of total BRSP and easily extractable BRSP were 0.95 mg g⁻¹ and 0.5 mg g⁻¹ in all data, respectively. The levels of BRSP in the desert soil were little lower than those in native and arable soils, but the ratios of BRSP to SOC were much higher than farmland soils. The results of this study support the conclusion that glomalin could be an appropriate index related to the level of soil fertility, especially in desert soil. Moreover, AM fungal colonizations and glomalin might be useful to monitor desertification and soil degradation.     

Effects of two soil reclamation techniques on the distribution of the organic N compounds in a 15N labelled burnt soil

The evolution of the soil organic-N forms and their bio-availability was studied in a ¹⁵N labelled and burnt soil (BLS) after two successive reclamation steps under greenhouse conditions: a 3-month growing period of Lolium, without (BLS-L) or with poultry manure addition (4 and 8 Mg ha^− 1: BLS + PM4-L and BLS + PM8-L), followed by a 12-month growing phase of pine seedlings (BLS-P, BLS + PM4-P and BLS + PM8-P). The results were compared with those obtained for the homologous labelled unburnt soil (LS, LS-L and LS-P) to evaluate the efficacy of these reclamation techniques in the mitigation of the drastic post-fire changes exhibited by the major biologically available N pool in terrestrial ecosystems: the soil organic N. The significant and steady decrease of the ¹⁵N enrichment observed in the unburnt soil during the successive plant growth cycles (LS > LS-L > LS-P) contrasts with the lack of significant changes, in both the content of total organic ¹⁵N and the atom % ¹⁵N in excess, among the treatments with the burnt soil (BLS ≥ BLS-L ≥ BLS-P). These results showed that: a) in LS, N mineralization proceeds faster for the recently incorporated N (¹⁵N enriched) than for the native N, supplying the growing vegetation with inorganic N more ¹⁵N enriched than the bulk soil N; and b) in BLS, soil combustion has reduced the usually higher biological availability of the recently added N to levels similar to those of the endogenous N.The re-vegetation with Lolium and Pinus and the addition of poultry manure mitigated the high differences observed in the size of the amino acid and the organic derived NH₄⁺–N pools due to the combustion process, which are usual between burnt and unburnt soils. Conversely, these burnt soil reclamation techniques (re-vegetation and poultry manure addition), even jointly used, were unable to reduce the huge differences observed between the burnt and the unburnt soils for the other N fractions considered (amides, amino sugars, hydrolysable unidentified-N, hydrolysable organic N and un-hydrolysable N) that accounted for more than 80% of the soil organic N. Consequently, it seems that without the introduction of N₂-fixing microorganisms or plants in the burnt soils the recovery of the natural soil organic N composition will take place slowly.     

Sorption of tannin and related phenolic compounds and effects on soluble-N in soil

Some tannins, plant-derived polyphenolic compounds, can rapidly affix to soil and affect the solubility of labile soil-N but a more complete understanding of the nature and persistence of tannin-soil interactions is needed. Forest and pasture soils from two depths were treated for 1 h with cool (23 ^°C) water (Control) or solutions that added 10 mg g⁻¹ soil tannic acid (TA), an imprecisely defined mixture of galloyl esters, gallic acid (GA), a phenol, or β-1,2,3,4,6-penta-O-galloyl-d-glucose (PGG), a hydrolyzable tannin. Soluble-C and N, in treatment supernatants, was measured to uncover evidence for sorption of treatments or effects on extraction of soil-N. Significant amounts of soluble-C, added with treatments, were not recovered in supernatants indicating sorption of nearly 90% of the PGG-C, about 75% of the TA-C but less than 25% of the GA-C in surface soil. Disappearance of soluble-C from treatment supernatants was accompanied by a corresponding reduction of total phenolic content. Treatments added a negligible amount of N to soil; but while PGG and TA reduced soluble-N, in extracts from surface soil, GA had little effect. Soluble-N in extracts was composed mainly of organic-N. Effects of tannins persisted in surface soil through 12 washings with hot water (80 ^°C), suggesting the formation of stable complexes with soil. The proportion of initial soil-C and N remaining after all extractions was higher in samples treated with PGG or TA than either the Control or GA treatment. We conclude PGG readily sorbs to soil and reduces the solubility of soil organic-N unlike GA, its simple monomeric constituent. These differences could be especially important near the surface where quantities of soil organic matter and biological activity are comparatively large and most easily affected by management.     

Impact of bovine urine deposition on soil microbial activity,biomass, and community structure

Nitrogen (N) from urine excreted by grazing animals can be transformed into N compounds that have detrimental effects on the environment. These include nitrate, which can cause eutrophication of waterways, and nitrous oxide, which is a greenhouse gas. Soil microbes mediate all of these N transformations, but the impact of urine on microbes and how initial soil conditions and urine chemical composition alter their responses to urine are not well understood. This study aimed to determine how soil inorganic N pools, nitrous oxide fluxes, soil microbial activity, biomass, and the community structure of bacteria containing amoA (nitrifiers), nirK, and nirS (denitrifiers) genes responded to the addition of urine over time. Bovine urine containing either a high (15.0 g K⁺ l^?1) or low salt content (10.4 g K⁺ l^?1) was added to soil cores at either low or high moisture content (hereafter termed dry and wet soil respectively; 35% or 70% water-filled pore space after the addition of urine). Changes in soil conditions, inorganic N pools, nitrous oxide fluxes, and the soil microbial community were then measured 1, 3, 8, 15, 29 and 44 days after urine addition. Urine addition increased soil ammonium concentrations by up to 2 mg g d.w.^?1, soil pH by up to 2.7 units, and electrical conductivity (EC) by 1.0 and 1.6 dS m^?1 in the low and high salt urine treatments respectively. In response, nitrate accumulation and nitrous oxide fluxes were lower in dry compared to wet urine-amended soils and slightly lower in high compared to low salt urine-amended soils. Nitrite concentrations were elevated (>3 μg g d.w.^?1) for at least 15 days after urine addition in wet urine-amended soils, but were only this high in the dry urine-amended soils for 1 day after the addition of urine. Microbial biomass was reduced by up to half in the wet urine-amended soils, but was largely unaffected in the dry urine-amended soils. Urine addition affected the community structure of ammonia-oxidising and nitrite-reducing bacteria; this response was also stronger and more persistent in wet than in dry urine-amended soils. Overall, the changes in soil conditions caused by the addition of urine interacted to influence microbial responses, indicating that the effect of urine on soil microbes is likely to be context-dependent.     

Relationships between soil pH and microbial properties in a UK arable soil

Effects of changing pH along a natural continuous gradient of a UK silty-loam soil were investigated. The site was a 200 m soil transect of the Hoosfield acid strip (Rothamsted Research, UK) which has grown continuous barley for more than 100 years. This experiment provides a remarkably uniform soil pH gradient, ranging from about pH 8.3 to 3.7. Soil total and organic C and the ratio: (soil organic C)/(soil total N) decreased due to decreasing plant C inputs as the soil pH declined. As expected, the CaCO₃ concentration was greatest at very high pH values (pH > 7.5). In contrast, extractable Al concentrations increased linearly (R² = 0.94, p < 0.001) from below about pH 5.4, while extractable Mn concentrations were largest at pH 4.4 and decreased at lower pHs. Biomass C and biomass ninhydrin-N were greatest above pH 7. There were statistically significant relationships between soil pH and biomass C (R² = 0.80, p < 0.001), biomass ninhydrin-N (R² = 0.90, p < 0.001), organic C (R² = 0.83, p < 0.001) and total N (R² = 0.83, p < 0.001), confirming the importance of soil organic matter and pH in stimulating microbial biomass growth. Soil CO₂ evolution increased as pH increased (R² = 0.97, p < 0.001). In contrast, the respiratory quotient (qCO₂) had the greatest values at either end of the pH range. This is almost certainly a response to stress caused by the low p. At the highest pH, both abiotic (from CaCO₃) and biotic Co₂ will be involved so the effects of high pH on biomass activity are confounded. Microbial biomass and microbial activity tended to stabilise at pH values between about 5 and 7 because the differences in organic C, total N and Al concentrations within this pH range were small. This work has established clear relationships between microbial biomass and microbial activity over an extremely wide soil pH range and within a single soil type. In contrast, most other studies have used soils of both different pH and soil type to make similar comparisons. In the latter case, the effects of soil pH on microbial properties are confounded with effects of different soil types, vegetation cover and local climatic conditions.     

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.     

Role of soil water content in the carbon and nitrogen dynamics of Lumbricus terrestris L. burrow soil

We evaluated the role of soil water content in controlling C and N dynamics within the drilosphere created by the anecic earthworm Lumbricus terrestris (L.). Mesocosms (volume = 3.1 l) were each amended with corn litter and three earthworms. Control treatments received no earthworms and no other earthworm species were present in the soil. WET and DRY treatments received a total of 9.25 cm and 3.25 cm of water, respectively. Water was added on weeks 1, 3, 7, and 10 at a rate of 2.0 cm per mesocosm for WET treatments and 0.5 cm per mesocosm for DRY treatments. Mesocosms were sampled destructively after incubation at 18–20 °C for 0, 3, 7, and 13 weeks. The water content of WET burrow soil ranged from 0.12 g g⁻¹ to 0.18 g g⁻¹ and was significantly higher than in the DRY treatment throughout the incubation period. The live weight of earthworms was significantly higher in the WET treatment only on week 13, whereas litter consumption was significantly lower in the DRY treatment for week 13. Carbon mineralization, measured as CO₂ evolved after a 24-h incubation, was consistently higher in WET than in DRY burrow soil. Effects of differences in soil water content were also apparent for biomass C and metabolic quotient. Soil water content did no affect the total C concentration of burrow soil. DRY burrow soil had consistently lower levels of nitrate than WET soil throughout the experiment. Lower levels of ammonium and inorganic N were observed for WET burrow soil on weeks 3 and 7. Water content did not have a significant effect on burrow soil total N. We concluded that the water content of the drilosphere affects both C and N dynamics and can affect the speciation of inorganic N; yet, the effects of soil water content do not appear to result from differences in the feeding activities of anecic earthworms.     

Mineralization dynamics and biochemical properties during initial decomposition of plant and animal residues in soil

The use of organic residues as soil amendments or fertilisers may represent a valuable recycling strategy. In this study, a series of laboratory assays was performed to study the effects of the application of organic residues on C and N mineralization and biochemical properties in a Mediterranean agricultural soil. Two crop residues (straw and cotton) and two animal by-products (meat bone meal and blood meal) were added at three rates (5, 10 and 20 mg g^?1 on dry weight basis) to a moist (40% water holding capacity) sandy soil and incubated at 20 °C for 28 days. Each residue underwent a different mineralization pattern depending on the nature and complexity of its chemical constituents. In all cases, the addition of the waste produced, after a short lag-phase, an exponential increase in the soil respiration rate, reflecting the growth of microbial biomass. The amount of total extra CO₂-C evolved after 28 days, expressed as % in respect to added C, differed significantly (P < 0.005) among application doses: 5 > 10 > 20 mg g^?1 and residue type: meat bone meal > blood meal > cotton cardings > wheat straw. Plant residues led to a rapid immobilisation of N that affected microbial size and activity and further mineralization. Animal by-products produced an immediate and remarkable increase of mineral N in the soil. However, the large amounts of NH₄⁺ released in the soil at high rates of animal residues led, in some cases, to temporary adverse effects on microbial biomass growth and nitrification. All residues produced a significant increase in soil microbial biomass size and activity, being the intensity of the response related to their chemical properties.     

Microbial mineralization of biochar and wheat straw mixture in soil: A short-term study

A short-term incubation study was carried out to investigate the effect of biochar addition to soil on CO₂ emissions, microbial biomass, soil soluble carbon (C) nitrogen (N) and nitrate–nitrogen (NO₃–N). Four soil treatments were investigated: soil only (control); soil + 5% biochar; soil + 0.5% wheat straw; soil + 5% biochar + 0.5% wheat straw. The biochar used was obtained from hardwood by pyrolysis at 500 °C. Periodic measurements of soil respiration, microbial biomass, soluble organic C, N and NO₃–N were performed throughout the experiment (84 days). Only 2.8% of the added biochar C was respired, whereas 56% of the added wheat straw C was decomposed. Total net CO₂ emitted by soil respiration suggested that wheat straw had no priming effect on biochar C decomposition. Moreover, wheat straw significantly increased microbial C and N and at the same time decreased soluble organic N. On the other hand, biochar did not influence microbial biomass nor soluble organic N. Thus it is possible to conclude that biochar was a very stable C source and could be an efficient, long-term strategy to sequester C in soils. Moreover, the addition of crop residues together with biochar could actively reduce the soil N leaching potential by means of N immobilization.     

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.     

Macronutrients and metals released from soils by solutions of naturally occurring phenols

Phenolic compounds produced by plants enter the soil by leaching and litter decomposition. The goal of this work is to determine the effect of phenolic compounds on solubility of plant macronutrients and metals in agroforestry systems. Soils from forest and pasture systems were repeatedly extracted with water (control) or phenolic solutions and then compared to a Mehlich 3 reference. The phenolics were aqueous solutions of tannic acid or β –1,2,3,4,6‐penta‐O‐galloyl‐D‐glucose (PGG) (hydrolyzable tannins), procyanidin (condensed tannin), or small phenolics catechin, gallic acid, or methyl gallate. The concentration of the macronutrients Ca, Mg, K, P, and S, and the metals Fe, Al, Mn, and Zn in the supernatants was determined by inductively‐coupled plasma spectroscopy. Cumulative extraction of macronutrients was generally similar to or less than the amount obtained by the Mehlich 3 extraction with the lowest recoveries obtained with the water control, PGG, and procyanidin. Metals tended to be somewhat more extractable from forest soil, especially with gallic acid, tannic acid or PGG treatments. Three mechanisms affected extraction of analytes by phenol‐containing solutions: (1) pH‐driven dissolution (Ca and Mg), (2) chelation of the metal (Al) by the polyphenol, or (3) reduction of the metal (Fe and Mn). Relatively low extraction of nutrients by some polyphenols is attributed to the tendency of some phenols to sorb to soil. This study demonstrates that tannins and related compounds change the solubility of macronutrients and metals in soils by a complex process that is not easily predictable from simple chemical properties of the phenolics.     

Impact of abundant Pheidole ant species on soil nutrients in relation to the food biology of the species

Impact of Pheidole sp., reportedly important in insect pest suppression in agroecosystems was studied on supporting agroecosystem services. This tropical ant species was found to be common and abundant in agroecosystems, with a high nest density and preference for the central, crop-growing zone of annual cropping systems. Physico-chemical characteristics of the debris soil were examined from nests located by the roadside and within two managed ecosystems. The debris soil had significantly higher concentrations of total C, N, P and NO₃-N along with higher water-holding capacity and moderate-sized soil particles in comparison to the control soil. The pH of the Pheidole sp. debris soil was shifted towards reduced alkaline conditions. Results reveal that annually, 2.44 kg/ha C, 0.071 kg/ha P, 0.628 kg/ha N and 0.009 kg/ha NO₃-N are added to the soil through the accumulation of organic refuse at the nest rim. This contributes to soil nutrient enhancement and is suggested to enhance ecosystem productivity. The high nutrient content of nest debris soil is linked to the predominance of arthropod carcasses (93.7% of the total organic refuse) in the refuse piles derived from the animal-based food (70.3%) brought to the nests by the foragers. Plant-based food was 29.6% (seeds, leaves, roots, etc.) of the total indicating a minor role of Pheidole sp. as a seed harvester. The results suggest an important role of Pheidole sp. in regulating the soil nutrients as an ecosystem engineer.     

Dynamic changes of bacterial community under the influence of bacterial-feeding nematodes grazing in prometryne contaminated soil

Microcosm experiments were carried out to study the effects of bacterial-feeding nematodes and prometryne on soil bacterial communities in contaminated soil. Prometryne (5 or 10 mg kg⁻¹ dry soil, that is, P5 or P10) and bacterial-feeding nematodes (5 or 10 individuals g⁻¹ dry soil, that is, N5 or N10), singly and in combination (P5N5, P5N10, P10N5, P10N10), were added to a nematode-free soil. An uncontaminated nematode-free soil was studied for comparison (Control). Bacterial-feeding nematode grazing boosted soil enzyme activities in contaminated soils, thus speeding up prometryne degradation. In the initial stage of the experiment, prometryne enhanced the soil enzyme activities too, but served the opposite purpose later. Denaturing gradient gel electrophoresis (DGGE) analysis indicated that prometryne contamination and nematode grazing over the incubation period exerted an obvious impact on Species richness (S), Shannon–Wiener index (H′) and Evenness (E_H) of soil bacteria, which increased initially, then decreased and increased again later. The cluster analysis of DGGE profiles showed that the similarity of soil bacterial communities in all treatments with indigenous microbes, P5, P5N5, P5N10, P10, P10N5, and P10N10 and the Control was 75%, 44%, 78% and 49% at Day 0, Day 8, Day 18 and Day 30, respectively. Compared to the Control, DGGE profiles displayed a varying characteristic bands pattern in all treatments over the incubation period with certain bands present in the treatments while not in the Control and vice versa, suggesting that bacterial-feeding nematode grazing and prometryne contamination affected soil bacterial communities evidently. Consequently, when added to contaminated soil, bacterial-feeding nematodes can contribute to restoration of contaminated sites by degrading toxic compounds like prometryne through enhanced microbial activity.     

Influence of carbon amendments on soil denitrifier abundance in soil microcosms

It is known that carbon (C) amendments increase microbial activity in anoxic soil microcosm studies, however the effects on abundance of total and denitrifier bacterial communities is uncertain. Quantitative PCR was used to target the 16S rRNA gene for the total bacterial community, the nosZ functional gene to reflect a broad denitrifier community, and functional genes from narrow denitrifier communities represented by Pseudomonas mandelii and related species (cnorB_P) and Bosea/Bradyrhizobium/Ensifer spp. (cnorB_B). Repacked soil cores were amended with varying amounts of glucose and red clover plant tissue (0–1000 mg C kg^? 1 of soil) and incubated for 96 h. Carbon amendment significantly increased respiration as measured by cumulative CO₂ emissions. Inputs of red clover or glucose at 1000 mg C kg^? 1 of soil caused increased abundance in the total bacteria under the conditions used. There was about an approximate 2-fold increase in the abundance of bacteria bearing the nosZ gene, but only in treatments receiving 500 or 1000 mg C kg^? 1 of soil of glucose or red clover, respectively. Additions of ≥ 500 mg C kg^? 1 soil of red clover and ≥ 250 mg C kg^? 1 of glucose increased cnorB_P-gene bearing denitrifiers. Changes in abundance of the targeted communities were related to C availability in soil, as indicated by soil respiration, regardless of C source. Applications of C amendments at rates that would occur in agricultural soils not only increase microbial activity, but can also induce changes in abundance of total bacterial and denitrifier communities in studies of anoxic soil microcosms.     

Changes in enzyme activities of spruce (Picea balfouriana) forest soil as related to burning in the eastern Qinghai-Tibetan Plateau

The effect of forest fire on soil enzyme activity of spruce (Picea balfouriana) forest in the eastern Qinghai-Tibetan Plateau was assessed. Six specific enzymes were chosen for investigation: invertase, acid phosphatase, proteinase, catalase, peroxidase and polyphenoloxidase. It was found that the activities of invertase and proteinase were reduced by burning, but the activities of acid phosphatase, polyphenoloxidase and peroxidase increased. Meanwhile, burning significantly (P < 0.05) resulted in the decrease of concentrations of available N and K of 0–20 cm depth layer soil, and significantly (P < 0.05) decreased concentrations of organic matter content, total N and P, as well as available N, P and K in soil at both 20–40 and 40–60 cm depths except for available P at 20–40 cm soil depth. These results illustrated that burning could influence the enzyme activities and chemical properties of soil not only of upper but also lower soil layers. Correlation analysis indicated that invertase activities in 0–20 cm depth layer soil were significantly positively correlated with organic matter, total N and P, as well as available N and P. Furthermore, all six enzymes studied were sensitive to fire disturbance, and thus could be used as indicators of soil quality. Our study also showed that soil enzyme activities were associated with soil depth, decreasing from top to bottom in both burned and unburned spruce forests. The distribution pattern of soil enzyme activities suggested that the rate of organic matter decomposition and nutrient cycling depended on soil depth, which had important structural and functional characteristics in nutrient cycling dynamics and implications in plantation nutrient management. The finding that burning effects on enzyme activities and soil properties between different soil layers were homogenized was attributed to the 8-years’ regeneration of forest after burning.     

Decoupling of microbial glucose uptake and mineralization in soil

The rate of organic matter turnover in soil is a critical component of the terrestrial carbon cycle and is frequently estimated from measurements of respiration. For estimates to be reliable requires that isotopically labelled substrate uptake into the soil microbial biomass and its subsequent mineralization occurs almost simultaneously (i.e. no time delay). Here we investigated this paradigm using glucose added to an agricultural soil. Immediately after collection from the field, various concentrations of ¹⁴C-labeled glucose (1 μM to 10 mM) were added to soil and the depletion from the soil solution measured at 1–60 min after substrate addition. ¹⁴CO₂ production from the mineralization of glucose was simultaneously measured. The microbial uptake of glucose from soil solution was concentration-dependent and kinetic analysis suggests the operation of at least two distinct glucose transport systems of differing affinity. At glucose concentrations reflecting those naturally present in the soil solution (54±10 μM), the half-time (t_1/2) of exogenous glucose was extremely rapid at ca. 30 s. At higher glucose concentrations (100 μM to 10 mM), the t_1/2 values for the high-affinity carrier were altered little, but increasing proportions of glucose were taken up by the low affinity transport system. Glucose mineralization by the soil microbial community showed a significant delay after its uptake into the microbial biomass suggesting a decoupling of glucose uptake and subsequent respiration, possibly by dilution of glucose in labile metabolite pools. By fitting a double first order kinetic equation to the mineralization results we estimated the t_1/2 for the first rapid phase of respiration at natural soil solution glucose concentrations to be 6–8 min, but at least 87% of the added glucose was retained in the microbial biomass prior to mineralization. Our results suggest that in this soil the soil solution glucose pool turns over 100–1000 times each day, an order of magnitude faster than when determined from measurements of mineralization. These results imply that traditional isotopic based measurements of substrate turnover measured using CO₂ may vastly underestimate their rate of cycling in soil.     

Nitrous oxide production in turfgrass systems: Effects of soil properties and grass clipping recycling

Soil N₂O emissions can affect global environments because N₂O is a potent greenhouse gas and ozone depletion substance. In the context of global warming, there is increasing concern over the emissions of N₂O from turfgrass systems. It is possible that management practices could be tailored to reduce emissions, but this would require a better understanding of factors controlling N₂O production. In the present study we evaluated the spatial variability of soil N₂O production and its correlation with soil physical, chemical and microbial properties. The impacts of grass clipping addition on soil N₂O production were also examined. Soil samples were collected from a chronosequence of three golf courses (10, 30, and 100-year-old) and incubated for 60 days at either 60% or 90% water filled-pore space (WFPS) with or without the addition of grass clippings or wheat straw. Both soil N₂O flux and soil inorganic N were measured periodically throughout the incubation. For unamended soils, cumulative soil N₂O production during the incubation ranged from 75 to 972 ng N g⁻¹ soil at 60% WFPS and from 76 to 8842 ng N g⁻¹ soil at 90% WFPS. Among all the soil physical, chemical and microbial properties examined, soil N₂O production showed the largest spatial variability with the coefficient of variation ~110% and 207% for 60% and 90% WFPS, respectively. At 60% WFPS, soil N₂O production was positively correlated with soil clay fraction (Pearson's r = 0.91, P < 0.01) and soil NH₄⁺–N (Pearson's r = 0.82, P < 0.01). At 90% WFPS, however, soil N₂O production appeared to be positively related to total soil C and N, but negatively related to soil pH. Addition of grass clippings and wheat straw did not consistently affect soil N₂O production across moisture treatments. Soil N₂O production at 60% WFPS was enhanced by the addition of grass clippings and unaffected by wheat straw (P < 0.05). In contrast, soil N₂O production at 90% WFPS was inhibited by the addition of wheat straw and little influenced by glass clippings (P < 0.05), except for soil samples with >2.5% organic C. Net N mineralization in soil samples with >2.5% organic C was similar between the two moisture regimes, suggesting that O₂ availability was greater than expected from 90% WFPS. Nonetheless, small and moderate changes in the percentage of clay fraction, soil organic matter content, and soil pH were found to be associated with large variations in soil N₂O production. Our study suggested that managing soil acidity via liming could substantially control soil N₂O production in turfgrass systems.