Is glomalin an appropriate indicator of forest soil reactive nitrogen status?

In this paper we address total glomalin‐related soil protein (T‐GRSP) as a possible indicator of differences in forest soils related to reactive nitrogen and forest composition. We focused especially on the relationship between T‐GRSP (g kg⁻¹), soil organic carbon (SOC), and reactive nitrogen (N_r) availability among different categories of temperate forests and different horizons. Our study included 105 sampling sites divided into 5 categories, which vary in elevation and tree species composition (coniferous, deciduous, mixed). We detected significantly higher T‐GRSP and SOC in the F+H horizon under conifers. We assume that this observation might be attributed to suppression of decomposition of T‐GRSP and SOC by nature of coniferous litter. The lack of significant differences in T‐GRSP/SOC among the categories and the positive correlations between T‐GRSP and SOC in most of the categories confirmed the strong relationship of T‐GRSP with SOC. We found a significantly higher content of T‐GRSP in the F+H horizon for all studied forest categories. However, the contribution of T‐GRSP to SOC is significantly higher in the A horizon, which might be caused by stabilization of glomalin by mineral fraction, including clay minerals or by the belowground origin of glomalin. We found the increase of SOC with increasing N_r in the A horizon for most categories of forest. T‐GRSP follows this trend in the case of deciduous forests (decid), mixed forest (mixed), and mountain forests (mount). On the other hand, we detected a decrease of T‐GRSP with increasing N_r in the F+H horizon of coniferous forests (conif). Moreover the T‐GRSP/SOC decreases with the increase of N_r in the A horizon of conif, mixed and mount, which points to the higher sensitivity of forest with prevalence of coniferous trees. Our observations have confirmed an ecosystem‐specific relationship between T‐GRSP, SOC and N_r. We concluded that T‐GRSP in combination with T‐GRSP/SOC has the potential to reveal qualitative changes in soil organic matter (SOM) connected with increasing N_r.     

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.     

Organic carbon and nitrogen dynamics in different soil fractions between broad-leaved Korean pine forests and aspen–birch forests in northeastern China

Purpose

Both overharvesting and climate changes have greatly altered forest composition in northeastern China; however, forest-specific effects on soil organic carbon (SOC), N, and compositional features in different soil fractions have not yet been defined.

Materials and methods

By sampling from broad-leaved Korean pine forest (the climax vegetation) and aspen–birch forest (the secondary forest), five soil fractions were separated by a physicochemical soil fractionation method, and Fourier transform infrared spectroscopy, X-ray diffraction analysis, and X-ray photoelectron spectrometry were used for functional groups, mineral diffraction, and elemental composition determination together with SOC and N measurements.

Results and discussion

Aspen–birch forests tended to sequestrate more SOC in the slow fractions (sand and aggregate [SA] and easily oxidized fractions) and more N in the sensitive fractions (particulate and soluble fractions), indicating that in aspen–birch forests, high SOC sequestration (1.26-fold) coincided with the active and rapid N supply. Much higher percentages (13.1–40.5 %) of O–H and N–H stretching, O–H bending, and C=O, COO–, and C–H stretching, and also the much lower quartz grain size and mineral diffraction peaks in SA and acid-insoluble fraction (over 85 % of total soil mass), in aspen–birch forests were possibly associated with the 1.17- to 1.53-fold higher SOC compared to broad-leaved Korean pine forest. However, elemental composition on soil particles might marginally contribute to the SOC and N forest-dependent differences.

Conclusions

Considering the increase of aspen–birch forests and the decrease of broad-leaved Korean pine forests in historical and future scenarios in northeastern China, more SOC has been and also will sequestrate in intact soils and stable soil fractions, with more N in sensitive fractions, and these should be highlighted in evaluating forest C and N dynamics during forest successions in this region.

     

Chemical characteristics of glomalin-related soil protein (GRSP) extracted from soils of varying organic matter content

Glomalin is described in the literature as a N-linked glycoprotein and the putative gene product of arbuscular mycorrhizal fungi (AMF). Since the link between glomalin and various protein fractions in soil is not yet clearly defined, glomalin-related soil protein (GRSP) more appropriately describes glomalin's existence in natural organic matter (NOM). The objective of this study was to examine the chemical characteristics of GRSP present in several mineral and organic soils of varying organic carbon content. GRSP was isolated using high temperature sodium citrate extraction followed by either trichloroacetic acid (TCA) or hydrochloric acid (HCl) precipitation. GRSP was characterized by quantitative solid-state ¹³C DPMAS NMR, infrared (IR) spectroscopy, elemental analysis, and the Bradford assay for protein content. GRSP accounted for 25% and 52% of total C in the mineral soils and organic soil, respectively. Molar C/N and H/C ratios reveal that GRSP has less nitrogen than bovine serum albumin (BSA), and that GRSP extracted from the Pahokee peat soil possessed a more unsaturated, and thus aromatic character relative to the mineral soil GRSP, respectively. GRSP's high aromatic (42-49%) and carboxyl (24-30%) carbon contents and low aliphatic (4-11%) and carbohydrate-type carbon contents (4-16%) suggests that GRSP does not resemble a typical glycoprotein. In fact, the NMR spectra of GRSP closely resemble that of humic acid. GRSP extracted from mineral and organic soils possessed the same NMR fingerprint regardless of the precipitation method used (i.e., either TCA or HCl). It is likely that the current GRSP extraction methods, because of their similarity to the method used to extract humic acid, are coextracting both materials.     

Arbuscular Mycorrhizal Fungi and Glomalin‐Related Soil Protein as Potential Indicators of Soil Quality in a Recuperation Gradient of the Atlantic Forest in Brazil

This study aimed at surveying arbuscular mycorrhizal fungi (AMF) species and glomalin‐related soil protein (GRSP) to understand their role as presumable biological indicators of soil quality in an undisturbed forest site (NT) and three sites with different management histories, soil textures, and different ages of recovery after reforestation for 20 (R20), 10 (R10) and 5 years (R05). Our objective was to determine how physical, chemical and microbiological soil attributes influence AMF species distribution, total‐GRSP (T‐GRSP) and easily extractable‐GRSP (EE‐GRSP). Glomus and Acaulospora were related to impacted sites, Gigaspora rosea to sites R10 and R20 that have different management histories and soil textures and Glomus geosporum to sites NT and R10, suggesting some influence of texture on its distribution. Scutellospora pellucida and other species were found only in one season. Correlations between EE‐GRSP and T‐GRSP on the one hand and total carbon and nitrogen, dehydrogenase and urease activity, microbial biomass carbon and microbial biomass nitrogen, on the other, reached values of 40–70% and were especially strong in summer. Soil bulk density had a negative and macroporosity a positive effect only on EE‐GRSP, suggesting the necessity to choose either EE‐GRSP or T‐GRSP as biological indicator depending on the soil characteristics and management. This study demonstrates the effect of recovery age, seasonality and other soil attributes on AMF and GRSP distribution and shows that these biological attributes may be used as indicators of soil quality in the Atlantic forest in Brazil. Copyright © 2013 John Wiley & Sons, Ltd.     

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.     

Characterization of glomalin as a hyphal wall component of arbuscular mycorrhizal fungi

Arbuscular mycorrhizal fungi (AMF) produce a protein, glomalin, quantified operationally in soils as glomalin-related soil protein (GRSP). GRSP concentrations in soil can range as high as several mg g⁻¹ soil, and GRSP is highly positively correlated with aggregate water stability. Given that AMF are obligate biotrophs (i.e. depending on host cells for their C supply), it is difficult to explain why apparently large amounts of glomalin would be produced and secreted actively into the soil, since the carbon could not be directly recaptured by the mycelium (and benefits to the AMF via increased soil structure would be diffuse and indirect). This apparent contradiction could be resolved by learning more about the pathway of delivery of glomalin into soil; namely, does this occur via secretion, or is glomalin tightly bound in the fungal walls and only released after hyphae are being degraded by the soil microbial community? In order to address this question, we grew the AMF Glomus intraradices in in vitro cultures and studied the release of glomalin from the mycelium and the accumulation of glomalin in the culture medium. Numerous protein-solubilizing treatments to release glomalin from the fungal mycelium were unsuccessful (including detergents, acid, base, solvents, and chaotropic agents), and the degree of harshness required to release the compound (autoclaving, enzymatic digestion) is consistent with the hypothesis that glomalin is tightly bound in hyphal and spore walls. Further, about 80% of glomalin (by weight) produced by the fungus was contained in hyphae and spores compared to that released into the culture medium, strongly suggesting that glomalin arrives mainly in soil via release from hyphae, and not primarily through secretion. These results point research on functions of glomalin and GRSP in a new direction, focusing on the contributions this protein makes to the living mycelium, rather than its role once it is released into the soil.     

Decomposition and the contribution of glomalin-related soil protein (GRSP) in heavy metal sequestration: Field experiment

Glomalin is a metal-sorbing glycoprotein excreted by arbuscular mycorrhizal fungi (AMF). One method of estimating glomalin in soils is as glomalin-related soil protein (GRSP). In this study the role of GRSP in sequestering Pb and Cd was investigated in an in situ field experiment. The effect of metal sequestration on the subsequent decomposition of GRSP was also investigated. GRSP was determined using the Bradford method as total glomalin-related soil protein (T-GRSP) and as easily extractable glomalin-related soil protein (EE-GRSP). After 140 days, GRSP bound Pb accounted for 0.21–1.78% of the total Pb, and GRSP bound Cd accounted for 0.38–0.98% of the total Cd content in the soil. However when compared on a soil organic matter (SOM) basis, only 4% of the Pb or Cd was bound to the GRSP fraction of the SOM compared with 40–54% of the Pb or Cd bound to the humin and fulvic acids in the SOM fraction. In soils contaminated with the highest levels of Pb and Cd, the T-GRSP (EE-GRSP) decomposition after 140 days was reduced by 8.0 (6.6)% and 7.0 (7.5)%, respectively, when compared with the controls. In the high Pb or Cd treatment groups we found that the fraction of metal bound to GRSP increased even though the total GRSP content declined over time. The mass ratio between Pb and GRSP-carbon changed from 2.3 to 271.4 mg (100 g)⁻¹ in all Pb levels soil, while with the high-Cd treatment group the mass ratio between Cd and GRSP-carbon (0.36 mg (100 g)⁻¹) was higher than the mass ratio seen with Cd-bound humic acid fractions. Our in situ field study shows that while GRSP does bind Pb and Cd, in the soils we investigated, the levels are insignificant compared to soil organic matter such as humic and fulvic acids.     

Re-examining the glomalin-purity of glomalin-related soil protein fractions through immunochemical,lectin-affinity and soil labelling experiments

Due to analytical similarities with the mycorrhizal glycoprotein glomalin, ubiquitous citrate and heat-extractable soil protein fractions have been assumed to be predominantly glomalin-stabilised within soil. Often termed glomalin-related soil protein (GRSP), little however is actually known of the “glomalin-purity” of these soil fractions. We undertook western and lectin blots and crossed immuno/lectin affinity electrophoresis (CIE/CLAE) analysis of “easily extractible” GRSP fractions, as well as liquid chromatography-tandem mass spectrometry (LC–MS/MS) of “total” GRSP fractions. To further test whether soil saprobes contribute to GRSP production, we amended soil with ¹⁴C-sucrose and examined whether ¹⁴C could be traced in the GRSP pool over a 500-day incubation period.While only four of six bands on SDS–PAGE profiles of easily extracted GRSP reacted with anti-glomalin MAb32B11 and the lectin Con A under our blotting conditions, CIE/CLAE indicated the presence of a single protein moiety in the easily extractible GRSP pool. LC–MS/MS analysis of total GRSP pooled from various soils also showed that although traces of protein tentatively assignable to soil bacteria were present in GRSP, their concentrations were low. Additionally, specific activity of total GRSP in ¹⁴C-labelled soil was relatively depleted compared to the bulk soil and soil microbial biomass. This suggests that little GRSP of heterotrophic origin was laid down over the incubation period, although the potential presence of a pre-existing ¹⁴C-free GRSP background, as well as of low microbial dynamics in the absence of any further substrate inputs to the soil warrant caution with this inference.     

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.     

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.     

Extraction of Glomalin and Associated Compounds with Two Chemical Solutions in Cultivated Tepetates of Mexico

Glomalin is a glycoprotein produced by the hyphae of arbuscular mycorrhizal fungi (AMF). The chemical methods usually employed to extract glomalin from the soil obtain something more than this pure glycoprotein, and therefore it would be better to call this fraction soil protein related to glomalin (SPRG) or glomalin associated with humic substances (GAHS). On this account, its isolation is controversial. The SPGR or GAHS has a significant influence on the physical, chemical, and biological properties of soils and could then be considered as an indicator of soil use change. In the present study, the storage of SPRG was evaluated, as well as carbon (C) associated with the latter (CG) and the content of soil organic C (SOC) in cultivated tepetates in the State of Mexico. Tepetates are hardened volcanic tuffs of the fragipan type, ameliorated for agricultural production. The specific objectives of the present study were (1) to evaluate the SPRG levels in tepetates, (2) measure the contribution of carbon (C) made by the SPRG to soil organic C (SOC), and (3) compare the extraction of SPRG with sodium pyrophosphate and sodium citrate. The samples used in this experiment came from 87 tepetate-cultivated plots (0–20 cm) located in the Texcoco River basin, State of Mexico. The levels of SPRG were observed among traces, 2.3 mg g^–1 for citrate and up to 5.6 mg g^–1 for pyrophosphate. The latter reactive allowed us to extract nearly three times more SPRG and two times more C-SPRG than sodium citrate (P?=?0.05) in the tepetates having about 4% of SOC; yet when the latter was less than 0.5%, the extraction levels with both solutions were similar. The SPRG and CG were closely correlated with SOC (r > 0.90). Tepetates have levels of SPRG similar to those observed in arid soils.     

Soil structural stability and erosion rates influenced by agricultural management practices in a semi‐arid Mediterranean agro‐ecosystem

Unsuitable agricultural practices can cause loss in soil quality and erodibility to thus increase or trigger desertification under Mediterranean conditions. A field experiment was performed at the El Teularet‐Sierra de Enguera Experimental Station (eastern Spain) to assess the influence during a 5‐yr period of different agricultural practices on physical and chemical indicators of soil quality (total and water‐soluble carbohydrates, glomalin‐related soil proteins (GRSP), total organic carbon, aggregate stability (AS), vegetation cover and soil erosion). The management practices included residual herbicide use, ploughing, ploughing + oats, addition of oat straw mulch and a control (land abandonment). Adjacent soil under natural vegetation was used as a reference for local, high‐quality soil and as a control for comparison with the agricultural soils under different management practices. Oat straw mulching led to higher levels of water‐soluble carbohydrates, GRSP and AS and lower soil erosion rates, resulting in values similar to those in the soil under native vegetation. The lowest levels of carbohydrates and GRSP were for the plots that were treated with herbicide or were ploughed. The maintenance of and increases in stable aggregates promoted by the different agricultural management practices over the years were attributed to increases in labile organic fractions such as carbohydrates and to the GRSP content. The results demonstrate that land abandonment (control plot) or the use of a cover (plants or straw) contributes to increases in soil quality and reduces the risk of erosion. The research also shows that sustainable agricultural management allows soil to recover and that the use of straw mulching is the most effective management strategy.     

No‐tillage with half‐amount residue retention enhances microbial functional diversity,enzyme activity and glomalin‐related soil protein content within soil aggregates

A 3‐year field tillage and residue management experiment established in North China was used to analyse topsoil (0–15 cm) aggregation, and microbial functional diversity, enzyme activity and glomalin‐related soil protein (GRSP) content within aggregates. Compared with conventional tillage (CT), no‐tillage (NT) alone significantly (P < 0.05) increased organic C contents in 50–250 and <2 μm aggregates and decreased the proportion of C accumulated by 2–50 μm aggregates and microbial functional diversity indices in <2 μm aggregates. Regardless of tillage practice, both half‐amount (C50) and full (C100) residue retention tended to increase organic C and GRSP contents, or dehydrogenase and invertase activities, in certain aggregates. Under CT, a poorer performance of C50 than C100 was observed in maintaining Shannon index (H′) and Simpson index (D) in >250 and <2 μm aggregates, and also McIntosh index (U) in <2 μm aggregates, owing to insufficient residue and possible decreases in the distribution of decomposer micro‐organisms. Under NT, however, C50 was more effective than C100 in maintaining/elevating H′, D and U in all soil aggregates except for 50–250 μm, suggesting that surplus residue may induce worse soil conditions, decreasing heterotrophic microbial activities. Thus, NT with half‐amount residue retention improved soil physical–chemical–biological properties and could be a useful management practice in North China.     

Effects of part and whole straw returning on soil carbon sequestration in C3–C4 rotation cropland

Long‐term no‐tillage management and crop residue amendments to soil were identified as an effective measure to increase soil organic carbon (SOC). The SOC content, SOC stock (SOCs), soil carbon sequestration rate (CSR), and carbon pool management index (CPMI) were measured. A stable isotopic approach was used to evaluate the contributions of wheat and maize residues to SOC at a long‐term experimental site. We hypothesized that under no‐tillage conditions, straw retention quantity would affect soil carbon sequestration differently in surface and deep soil, and the contribution of C3 and C4 crops to soil carbon sequestration would be different. This study involved four maize straw returning treatments, which included no maize straw returning (NT‐0), 0.5 m (from the soil surface) maize straw returning (NT‐0.5), 1 m maize straw returning (NT‐1), and whole maize straw returning (NT‐W). The results showed that in the 0–20 cm soil layer, the SOC content, SOCs, CSR and CPMI of the NT‐W were highest after 14 years of no‐tillage management, and there were obvious differences among the four treatments. However, the SOC, SOCs, and CSR of the NT‐0.5 and NT‐W were the highest and lowest in 20–100 cm, respectively. The value of δ¹³C showed an obviously vertical variability that ranged from –22.01‰ (NT‐1) in the 0–20 cm layer to –18.27‰ (NT‐0.5) in the 60–80 cm layer, with enriched δ¹³C in the 60–80 cm (NT‐0.5 and NT‐1) and 80–100 cm (NT‐0 and NT‐W) layers. The contributions of the wheat and maize‐derived SOC of the NT‐0.5, NT‐1 and NT‐W increased by 11.4, 29.5 and 56.3% and by 10.7, 15.1 and 40.1%, relative to those in the NT‐0 treatment in the 0–20 cm soil layer, respectively. In conclusion, there was no apparent difference in total SOC sequestration between the NT‐0.5, NT‐1, and NT‐W treatments in the 0–100 cm soil layer. The contribution of wheat‐derived SOC was higher than that of maize‐derived SOC.     

Investigating the effects of wettability and pore size distribution on aggregate stability: the role of soil organic matter and the humic fraction

Soil organic matter (SOM) is an important factor influencing aggregate stability. Interactions between SOM and soil structure are widely studied, although the subtle relationship between SOM content, pore size distribution and aggregate stability is not fully known. Here we investigate such a relationship by means of a long‐term experiment established in 1962 in northeastern Italy, which considers different fertilizer practices (organic, mineral and mixed) applied to a continuous maize crop rotation. We measured wet stability of 1–2 mm aggregates subjected to different pretreatments. Both soil physical properties (such as pore size distribution and hydrophobicity) and chemical properties (soil organic and humic carbon content) affecting aggregate stability were considered. The chemical structure of humic substances was characterized by thermal and spectroscopic analyses (TG‐DTA, DRIFT and ¹H HR MAS NMR). The Pore‐Cor network model was then applied to evaluate the contribution of hydrophobicity and porosity to aggregate wetting. Our study suggests that SOM and its humic fraction can affect aggregate wetting and consequently slaking by modifying the pore size distribution with a shift from micropores (5–30 µm) and mesopores (30–75 µm) to ultramicropores (0.1–5 µm); hydrophobicity was also increased as a result of different humic composition. Spectroscopic analysis showed that hydrophobic compounds were mostly associated with complex humic molecules. Models of fast wetting dynamics, however, suggest that the contribution that hydrophobicity makes to aggregate stability, especially to soils with large carbon inputs, may not be the most significant factor.     

Decreased glomalin-related soil protein with nitrogen deposition in a 3-year-old Cunninghamia lanceolata plantation

Purpose

Glomalin-related soil protein (GRSP) is an essential component of soil organic C for maintaining soil quality and structure and plays a critical role in soil carbon (C) sequestration. However, how GRSP changes under nitrogen (N) deposition remains poorly understood.

Materials and methods

We assessed total GRSP (T-GRSP) and easily extractable GRSP (EE-GRSP) under a control (no N input), low N addition (LN, 40 kg N ha^?1 year^?1), and high N addition (HN, 80 kg N ha^?1 year^?1) treatments in 2015 and 2016 in a Chinese fir (Cunninghamia lanceolata) plantation in the subtropical China. We also analyzed soil properties contents and explored the stoichiometric ratios of soil organic C (SOC), total N (TN), and total phosphorus (TP) with GRSPs.

Results

Compared to the control, both T-GRSP and EE-GRSP were significantly reduced under the HN treatment, but had no significant difference under the LN treatment. The ratio of T-GRSP and EE-GRSP was reduced by the N addition. Soil organic C (SOC) and dissolved organic C (DOC) were significantly affected by N addition treatments. The ratios of GRSP-C to SOC and of EEGRSP-C to SOC ranged from 6.29 to 16.07% and 1.34 to 3.52%, respectively. T-GRSP and EE-GRSP were positively correlated with SOC/TN ratio, but negatively correlated with soil TN/TP and SOC/TP ratios.

Conclusion

Our results indicated that the GRSP reductions under N deposition in soil are mediated by soil C, N, and P stoichiometry, and particularly, the reduction of EE-GRSP by DOC. This study improved our mechanistic understanding of dynamics of GRSPs under increasing N enrichment in subtropical plantation ecosystems.

     

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.     

Straw mulching with fertilizer nitrogen: An approach for improving crop yield,soil nutrients and enzyme activities

Field experiments were conducted to study soil properties, soil enzymes activities, water use efficiency (WUE) and crop productivity after six years of soya bean straw mulching in the semi‐arid conditions of China. The experiment included four treatments: CK (Control), N (240 kg N ha^‐1), H (soya bean straw mulching at half rate 700 kg ha^‐1 with 240 kg N ha^‐1) and F (soya bean straw mulching at full rate 1,400 kg ha^‐1 with 240 kg N ha^‐1). Soil organic carbon (SOC), soil labile organic carbon (LOC), soil available N (AN), available P (AP) and enzyme activities were analysed after wheat harvesting in 2016 and 2017. Results show that straw amounts had positive effects on the soil fertility indices being higher for treatment F. The SOC, LOC, AN, AP and enzyme activities (i.e. saccharase, urease and alkaline phosphatase) were in the order of F > H > N > CK. High wheat grain yield and WUE were observed for F treatment. A total of six years mulching along with 240 kg ha^‐1 nitrogen fertilizer application is sufficient for wheat yield stability and improving soil properties except urease activities in the semi‐arid condition of China. However, the straw mulching amount should be further studied with minimum nitrogen fertilizer for an environment‐friendly and effective approach for improving the soil biological properties with adequate crop production on a sustainable basis in the semi‐arid region of China.