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.     

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.     

Glomalin-related soil protein and water relations in mycorrhizal citrus (Citrus tangerina) during soil water deficit

Glomalin-related soil protein (GRSP), a glycoprotein of arbuscular mycorrhizal fungi (AMF) secreted into soil, governs the aggregate stability, but the role of GRSP in soil and plant water is sparsely studied. The 24-week-old red tangerine (Citrus tangerina) inoculated with Glomus etunicatum and G. mosseae were subjected to a soil drying for 12 days as soil water deficit (SWD). Length of SWD significantly reduced mycorrhizal colonization, soil hyphal length, and leaf and soil water potential (Ψ), but increased total GRSP (T-GRSP), easily extractable GRSP (EE-GRSP), and proportion of water-stable aggregates (WSAs) in >0.25 mm size, irrespective of AMF source. The AMF-inoculated seedlings showed significantly higher T-GRSP, EE-GRSP, and leaf/soil Ψ than non-AMF seedlings during SWD. A significantly positive correlation was observed for mycorrhizal colonization versus leaf or soil Ψ, and hyphal length versus leaf Ψ, suggesting that root intra- and extra-radical hyphae participated in water transport. Interestingly, in GRSP fractions, only T-GRSP was significantly positively correlated with 0.25–1 and >0.25 mm WSA and negatively with leaf and soil Ψ. These results revealed a strong glue function of T-GRSP (not EE-GRSP and hyphae) to alter the proportional distribution of WSA size, thereby aiding toward prevention of soil water loss for improving soil–plant water relations.     

Plant invasion of native grassland on serpentine soils has no major effects upon selected physical and biological properties

Plant invasions alter soil microbial community composition; this study examined whether invasion-induced changes in the soil microbial community were reflected in soil aggregation, an ecosystem property strongly influenced by microorganisms. Soil aggregation is regulated by many biological factors including roots, arbuscular mycorrhizal fungal hyphae, and microbially-derived carbon compounds. We measured root biomass, fungal-derived glomalin-related soil protein (GRSP), and aggregate mean weight diameter in serpentine soils dominated by an invasive plant (Aegilops triuncialis (goatgrass) or Centaurea solstitialis (yellow starthistle)), or by native plants (Lasthenia californica and Plantago erecta, or Hemizonia congesta). Root biomass tended to increase in invaded soils. GRSP concentrations were lower in goatgrass-dominated soils than native soils. In contrast, starthistle dominated soil contained a higher amount of one fraction of GRSP, easily extractable immunoreactive soil protein (EE-IRSP) and a lower amount of another GRSP fraction, easily extractible Bradford reactive soil protein (EE-BRSP). Soil aggregation increased with goatgrass invasion, but did not increase with starthistle invasion. In highly aggregated serpentine soils, small increases in soil aggregation accompanying plant invasion were not related to changes in GRSP and likely have limited ecological significance.     

Losses of glomalin-related soil protein under prolonged arable cropping: A chronosequence study in sandy soils of the South African Highveld

Residues of arbuscular mycorrhizal fungi (AMF) may be important for agroecosystem functioning due to their ability to promote soil aggregation, especially in coarse textured soils with little biomass input and low capacity to conserve soil organic matter (SOM). Our aim was to assess the fate of AMF residues with prolonged arable cropping in coarse textured soils in a subtropical savannah assuming that glomalin-related soil protein (GRSP), especially the MAb32B11-immunoreactive fraction, mainly constitutes material of AMF origin. In three agroecosystems on the South African Highveld, surface soils were sampled. The former grassland soils had a history of up to 98 yr of cropping. We measured four GRSP fractions: Bradford-reactive soil protein (BRSP) and immunoreactive soil protein (IRSP), and easily extractable fractions of both. The primary grassland sites exhibited generally low contents of SOM and low GRSP contents. Prolonged arable land use of former grassland soils reduced the content of GRSP further. The decline could be described with a mono-exponential function with rate constants ranging from 0.04 to 0.41 yr⁻¹. Depending on the GRSP fraction, steady-state conditions were reached after 11-92 yr on a level of 39-69% of the initial contents. We conclude that even though GRSP fractions had the same hypothesized origin, they comprised pools with different stability or replacement rate. Easily extractable IRSP was lost most rapidly. In contrast to carbon, nitrogen and microbial residue dynamics, GRSP contents were not reduced below a certain steady-state level, despite potentially negative management effects on AMF, such as tillage, inclusion of fallows into crop rotation and fertilization with inorganic phosphorus. The steady-state GRSP contents coincided with low, but steady agroecosystem yields under the given cropping management.     

Arbuscular mycorrhizal fungi contribute to C and N enrichment of soil organic matter in forest soils

Increasing evidence suggests that accretion of microbial turnover products is an important driver for isotopic carbon (C) and nitrogen (N) enrichment of soil organic matter (SOM). However, the exact contribution of arbuscular mycorrhizal fungi (AMF) to soil isotopic patterns remains unknown. In this study, we compared ¹³C and ¹⁵N patterns of glomalin-related soil protein (GRSP), which includes a main fraction derived from AMF, litter, and bulk soil in four temperate rainforests. GRSP was an abundant C and N pool in these forest soils, showing significant ¹³C and ¹⁵N enrichment relative to litter and bulk soil. Hence, cumulative accumulation of recalcitrant AMF turnover products in the soil profile likely contributes to ¹³C and ¹⁵N enrichment in forest soils. Further research on the relationship between GRSP and AMF should clarify the exact extent of this process.     

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.     

Mycorrhizal function on soil aggregate stability in root zone and root-free hyphae zone of trifoliate orange

Arbuscular mycorrhizal fungi (AMF) influence soil aggregate stability through their hyphae, roots, and glomalin-related soil protein (GRSP); however, the individual effect of these factors is difficult to distinguish. Pots separated by a 37-μm mesh bag buried in the middle of each pot was used to establish root zone (root + hyphae) and hyphae zone (roots free), where the Poncirus trifoliata seedlings were colonized by Funneliformis mosseae or Paraglomus occultum in root zone. AMF inoculation significantly increased shoot, root, and plant’s total biomass, soil organic carbon, GRSP fractions, 2–4 and 1–2 mm size water-stable aggregates, and mean weight diameter (MWD) in root or hyphae zone. Within root zone, root colonization and biomass presented stronger relationship with MWD than GRSP fractions. While, within hyphae zone, total of GRSP fraction was significantly correlated with MWD. The study, suggested further that root biomass and colonization were the main mechanisms in root zone for improving aggregate stability, whereas total of GRSP fractions was of paramount importance in hyphae zone. Mycorrhizal effect on aggregate stability was observed to be contrastingly different between root zone and hyphae zone.     

Glomalin-related soil protein responses to elevated CO2 and nitrogen addition in a subtropical forest: Potential consequences for soil carbon accumulation

According to the economy theory, plants should preferentially allocate photosynthate to acquire below-ground resources under elevated atmospheric carbon dioxide (eCO₂) but decrease below-ground C allocation when nitrogen (N) is sufficient for plant growth. Arbuscular mycorrhizae (AM) represent a critical mechanism of below-ground nutrient acquisition for plants. The dynamics of arbuscular mycorrhizal fungi (AMF) could therefore reflect the response of plant C allocation under eCO₂ and N addition. We examined the responses of glomalin-related soil protein (GRSP) to eCO₂ (approximately 700 μmol mol⁻¹ CO₂) and/or N addition (100 kg N ha⁻¹ yr⁻¹ as NH₄NO₃) in a modeled subtropical forest to better understand its potential influence on soil C storage. We hypothesized that GRSP would increase under eCO₂ and decrease under N addition. Furthermore, the positive effects of eCO₂ on GRSP would be offset by extra N addition, and GRSP would remain unchanged under combined eCO₂ and N addition. Our results showed that the mean concentrations of easily extractable GRSP (EE-GRSP) and total GRSP (T-GRSP) were 0.35 ± 0.05 and 0.72 ± 0.13 mg C cm⁻³, respectively, which accounted for 2.76 ± 0.53% and 5.67 ± 0.92% of soil organic carbon (SOC) in the 0–10 cm soil layer. Elevated CO₂ significantly increased T-GRSP by 35.02% but decreased EE-GRSP by 5.09% in the top 10 cm soil layer. The opposite responses of T-GRSP and EE-GRSP to eCO₂ might result from an unchanged photosynthate investment to AMF with possible changes in their decomposition rates. The effect of N on GRSP was contrary to our hypothesis, i.e., there was a 1.72%–48.49% increase in T-GRSP and a slightly increase in EE-GRSP. Both EE-GRSP and T-GRSP concentrations increased under the combination of eCO₂ and N addition, which was inconsistent with our hypothesis. The significant increase of EE-GRSP under the combination of eCO₂ and N addition was partly caused by more rapid plant growth and reduced microbial diversity, and the marginal increase of T-GRSP indicated that the interaction between eCO₂ and N addition offset their independent effects. In addition, the relatively higher accumulation ratios of GRSP (22.6 ± 13.6%) compared with SOC (15.9 ± 9.4%) indicated that more rapid GRSP deposition in the soil might accelerate SOC accumulation under eCO₂ and N addition. Our results will improve the understanding of the functioning of GRSP in soil C sequestration under global environmental change scenarios.     

Priming effects in soil size fractions of a podzol Bs horizon after addition of fructose and alanine

It is well known that the addition of easily available substrates to soils can affect microbial activity and thus the mineralization of soil organic carbon (SOC). Up to now, little is known about the processes leading to these priming effects and which fractions of organic matter (OM) are affected. The objectives of this study were to determine if SOC associated with isolated soil size fractions showed different susceptibility to priming effects, whether these pools are easily depleted, or whether the amount of substrate addition affects the extent of priming effects. In an incubation experiment, the effect of the uniformly ¹⁴C‐labeled substrates fructose and alanine on the mineralization of the SOC of a Bs horizon of a Haplic Podzol was investigated. The soil sample was fractionated into the three soil size fractions sand, silt, and clay by a mild sonication followed by sieving and sedimentation. Additionally, nonfractionated soil of the horizon was included in the experiment. Every soil sample received four substrate additions repeated at weekly intervals with 3.325 μg substrate‐C (mg SOC)^–1 and a final addition of 13.3 μg substrate‐C (mg SOC)^–1 after 4 weeks. The respiration was determined hourly and ¹⁴CO₂ was analyzed every 2, 4, and 7 d after the respective substrate addition. After 56 d, between 42% and 58% of the added substrates had been mineralized. Both substrates strongly increased the mineralization of the OM in all fractions (positive priming effects). The priming effects were always higher after the addition of the high substrate dose than during the first 4 weeks when four small doses were added. In general, the priming effects increased with decreasing particle size. Alanine generally caused higher priming effects than fructose in the soil size fractions (up to 280% vs. 231%, respectively). This indicates that alanine serves not only as an energy substrate but also as a N source and, thus, also promotes microbial growth. The strong priming effects in the silt and clay fraction (133% and 125% with fructose, 172% and 168% with alanine) showed, that not only the labile pool of OM is affected, but also a more stable pool characterized by higher ¹⁴C ages. We assume that the stability of the OM in these fractions is not only due to recalcitrance or to interactions with the minerals, but that it may also be caused by a substrate limitation of the degrading microorganisms.     

Impacts of land use change on soil aggregation and aggregate stabilizing compounds as dependent on time

Land use change is known to strongly affect soil aggregation and aggregate stabilizing compounds. In this study we wanted to gain insight into the temporal and spatial dimension of this process. Therefore, we studied water-stable aggregates, total organic carbon (TOC), carbohydrates and glomalin-related soil protein (GRSP) in a 110-year chronosequence of Stagnosols, which have been converted from pasture to cropland at different times in history. To describe the temporal dimension, the measured concentrations were approximated by an exponential decay function. The spatial dimension was assessed by analyzing the distribution of TOC, carbohydrates, GRSP, and ¹⁴C among the different aggregate-size fractions over the course of 110 years of cropland use. It was found that the TOC concentration decreased monoexponentially (R² = 0.92) from 195.2 t ha⁻¹ to 45.13 t ha⁻¹ in the first depth interval (0-20 cm) during the first 110 years after the conversion, and reached a new equilibrium 23 (±5) years after land use change. Carbohydrates and GRSP obtained a new equilibrium after 14 (±6) and 56 (±5) years in the same depth interval. The mean-weight diameter (MWD) of the water-stable aggregates reached a new equilibrium 33 (±2) years after the land use conversion. With respect to the spatial dimension we found that TOC, carbohydrates, and GRSP showed higher concentrations in the macroaggregates than in the microaggregates. The ratios of the distribution of TOC, carbohydrates and GRSP among macro- and microaggregates did not change significantly during the 110 years of arable use of the soils. The average age of the organic carbon in the different aggregate-size fractions analyzed by its ¹⁴C concentration showed of a wide range from 65 (±25) to 251 (±30) years, and did not change significantly during the cultivation of the sites. Thus, we conclude that water-stable aggregates and the concentrations of TOC, carbohydrates and GRSP reacted towards land use change at different speeds and reached a new equilibrium between 14 (±6) and 56 (±5) years after the conversion of land use. Secondly, we found, that the spatial distribution of aggregate stabilizing compounds was not significantly changed during the first 110 years of cultivation.     

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.

     

Effect of superphosphate applications on soil phosphorus and on crop phosphorus removal by the strawberry

Abstract

Rate of superphosphate application significantly increased the Bray #2 (easily acid soluble plus adsorbed P) and “AlPO₄”; fractions for 3 successive years. A single dolomitic limestone application increased “FePO₄”; by an average of 36% one year after application. Crop P removal by the strawberry cv. ‘Acadia’ was approximately 2–3 ppm each year and it was not possible to relate this small amount to any of the soil P fractions determined.     

Arbuscular mycorrhizal-like fungi and glomalin-related soil protein drive the distributions of carbon and nitrogen in a large scale

Purpose

Arbuscular mycorrhizal-like fungi (AM-like fungi) are crucial for ecosystem functioning and soil organic matter (SOM) is an indicator of soil quality. However, the spatial distribution of arbuscular mycorrhizal-like fungi, glomalin-related soil protein (GRSP) and SOM in a large scale is still unclear. The objectives of this study were to investigate the spatial distribution of SOM, arbuscular mycorrhizal-like fungi and GRSP, and reveal the potential relationship among them in a large scale across China.

Materials and methods

Soil samples (different in vegetation type, climate, and soil variables) were collected from 26 sites in a large scale across China. The soil properties including pH, total carbon (TC), total nitrogen (TN), and SOM were determined. Quantitative PCR amplification of the 18S rRNA gene was conducted to evaluate the abundance of arbuscular mycorrhizal-like fungi. The contents of easily extractable GRSP (EE-GRSP), difficultly extractable GRSP (DE-GRSP), and total GRSP (T-GRSP) were measured.

Results and discussion

Arbuscular mycorrhizal-like fungi abundance was significantly affected by the vegetation type and dramatically correlated with the soil TN and mean annual precipitation (MAP). EE-GRSP and DE-GRSP were more associated with the TC and TN content, respectively. The abundance of arbuscular mycorrhizal-like fungi significantly but weakly correlated with the T-GRSP and EE-GRSP. The SOM content positively correlated with the DE-GRSP and T-GRSP. Those results suggested that the arbuscular mycorrhizal-like fungi are a larger contributor to regulating the content of GRSP, which is an important indicator of the soil organic carbon pool.

Conclusions

Our results indicated that arbuscular mycorrhizal-like fungi abundance has a greater contribution to driving the distribution of soil C and N in a large scale by affecting the content of glomalin-related soil protein.

     

Glomalin amount and compositional variation,and their associations with soil properties in farmland,northeastern China

Glomalin‐related soil protein (GRSP) is well‐known for its soil conditioning functions, but compositional traits are rarely considered. Farmland in northeastern China is the most important commercial grain basis, and soil degradation becomes the bottleneck for keeping crop productivity. The objective of this study was to uncover the possible associations between GRSP (amount and composition) and soil properties, and make suggestions for soil improvement from soil glomalin rehabilitation in northeastern China. Here, spatial variation in GRSP amount (Easily‐extractable‐GRSP, EE‐GRSP; Total‐GRSP, T‐GRSP) and its compositional traits from infrared spectroscopy, UV‐absorbance, X‐ray diffraction (XRD) and 3‐D fluorescence spectroscopy were surveyed in 360 soil samples across northeastern China, and their association with 11 soil properties were also analyzed for finding the possible influence of soil properties on GRSP composition in farmland. There about 3‐fold spatial variation in GRSP amount was observed, while functional group variations were ranged from 1.2‐fold (O–H & N–H stretching) to 2.4‐fold (C–O stretching & O–H bending of –COOH) in different locations. The XRD showed that grain size was 113–180Å and crystallinity was 0.71–1.42%, and GRSP contained seven fluorescent compounds of tyrosine‐like, tryptophan‐like, fulvic acid‐like, soluble microbial byproduct, humic acid‐like, nitrobenzoxadidole‐like, and calcofluor white‐like. Both, EE‐GRSP and T‐GRSP positively associated with soil organic carbon (SOC), soil N (SON), soil P (SOP), alkali‐hydrolyzed N (AN), available P (AP), available K (AK), and soil water, while negatively associated with soil pH and soil bulk density. Structural equation model (SEM) analysis indicates that direct effects on GRSP amounts were mainly from soil bulk density (coefficient: –0.27), soil pH (coefficients: –0.51 to –0.57), SOC (coefficients: 0.51 to 0.69) and AP (coefficients: 0.18 to 0.26), while all other soil properties had indirect effects on GRSP amounts via their close associations with these four parameters. Compared with the GRSP amounts, soil properties laid fewer effects on GRSP compositional traits. Of 16 compositional traits, five of them showed possible regulations from soil properties, which were three infrared functional groups (IR‐II: aliphatic C–H stretching; IR‐V: C–O stretching & O–H bending of –COOH; IR‐VII: O–H binding) and two fluorescent compounds (tyrosine‐like and humic acid‐like). SEM analysis indicates that soil water, pH and EC could directly affect IR‐II, IRV, tyrosine‐like and humic acid‐like, while available nutrients showed more evident influences on infra‐red functional groups than total amounts of N, P and K. Moreover, SOC, as a media of various soil nutrients, gave the strongest influence on GRSP compositional traits. As a supplement to previous studies, we found that GRSP is a mixture of different fluorescent compounds with different functional groups. Our findings highlight that soil properties could strongly change both GRSP accumulation in soil and their compositional traits, and the definition of the most probable soil properties in regulating glomalin amount and composition in this paper could favor good soil management in farmland at northeastern China.     

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.     

Characterization of recently 14C pulse-labelled carbon from roots by fractionation of soil organic matter

The inability of physical and chemical techniques to separate soil organic matter into fractions that have distinct turnover rates has hampered our understanding of carbon (C) and nutrient dynamics in soil. A series of soil organic matter fractionation techniques (chemical and physical) were evaluated for their ability to distinguish a potentially labile C pool, that is ‘recent’ root and root‐derived soil C. ‘Recent’ root and root‐derived C was operationally defined as root and soil C labelled by ¹⁴CO₂ pulse labelling of rye grass–clover pasture growing on undisturbed cores of soil. Most (50–94%) of total soil + root ¹⁴C activity was recovered in roots. Sequential extraction of the soil + roots with resin, 0.1 m NaOH and 1 m NaOH allocated ‘recent’ soil + root ¹⁴C to all fractions including the alkali‐insoluble residual fraction. Approximately 50% was measured in the alkali‐insoluble residue but specific activity was greater in the resin and 1 m NaOH fractions. Hot 0.5 m H₂SO₄ hydrolysed 80% of the ¹⁴C in the alkali‐insoluble residue of soil + roots but this diminished specific activity by recovering much non‐¹⁴C organic matter. Pre‐alkali extraction treatment with 30% H₂O₂ and post‐alkali treatment extractions with hot 1 m HNO₃ removed organic matter with a large ¹⁴C specific activity from the alkali‐insoluble residue. Density separation failed to isolate a significant pool of ‘recent’ root‐derived ¹⁴C. The density separation of ¹⁴C‐labelled roots, and roots remixed with non‐radioactive soil, showed that the adhesion of soil particles to young ¹⁴C‐labelled roots was the likely cause of the greater proportion of ¹⁴C in the heavy fraction. Simple chemical or density fractionations of C appear unsuitable for characterizing ‘recent’ root‐derived C into fractions that can be designated labile C (short turnover time).     

Development of liquid chromatography mass spectrometry method for analysis of organic N monomers in soil

The aim of this study was to develop an analytical procedure based on liquid chromatography-mass spectrometry (LC–MS) for analysis of monomeric organic N compounds in soil extracts. To benchmark the developed LC–MS method it was compared with a capillary electrophoresis–mass spectrometry (CE–MS) method recently used for analysis of small organic N monomers in soil. The separation was optimized and analytical performance assessed with 69 purified standards, then the LC–MS method was used to analyse soil extracts. Sixty-two out of 69 standards were analysable by LC–MS with separation on a hydrophilic interaction liquid chromatography column. The seven compounds that could not be analysed were strongly cationic polyamines. Limits of detection for a 5 μL injection ranged between 0.002 and 0.38 μmol L⁻¹, with the majority (49 out of 62) having limits of detection better than 0.05 μmol L⁻¹. The overall profile and concentration of small organic N monomers in soil extracts was broadly similar between LC–MS and CE–MS, with the notable exception of four ureides that were detected by LC–MS only. In soil extracts that had been concentrated ten-fold the detection and quantification of (some) organic N compounds was compromised by the presence of large amounts of inorganic salts. The developed LC–MS method offered advantages and disadvantages relative to CE–MS, and a combination of the two methods would achieve the broadest possible coverage of organic N in soil extracts.     

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.     

Water-stable aggregates, glomalin-related soil protein, and carbohydrates in a chronosequence of sandy hydromorphic soils

The conversion of pasture to cropland leads to a decline of aggregation in topsoils and to a decrease of aggregate-binding agents such as carbohydrates and glomalin-related soil protein (GRSP). Till now, studies on soil aggregation focused either on carbohydrates or on GRSP as a binding agent in aggregates. In this study we analyse the development of the relationship between carbohydrates, GRSP, TOC and aggregate-stability following land-use change. Furthermore, we discuss the contents of carbohydrates, GRSP and TOC in each of the aggregate fractions. For these purposes, a chronosequence of sites, which were converted from pasture to cropland at different periods in history, was established. To get further insight into the impact of different types of land-use, also soils under forest, either afforested or permanent, were studied. The mean-weight diameter (MWD) of water-stable aggregates, the carbohydrate, and the GRSP content were determined in 49 soils. It was found that the MWD of the water-stable aggregates decreased monoexponentially (R² = 0.66) by 66% during the first 46 years after conversion of the soils from pasture to cropland. During the same period, the carbohydrate content decreased very rapidly after the land use change by 64% and the GRSP content decreased more slowly by 57%. The MWD of the forest soils were in the same range as those of the permanent pasture soils although they exhibit significantly higher TOC contents, which indicate that other stabilization mechanisms are dominant in forest soils, less important in the chronosequence soils. TOC, carbohydrates and the GRSP contents were sigmoidally correlated with the MWD. Among the four water-stable aggregate fractions TOC and carbohydrates exhibited high contents in the macroaggregates and were less present in the microaggregates. GRSP, in contrast, was more equally distributed among the four water-stable aggregate fractions.