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Glomalin-related soil protein (GRSP) has demonstrated significant potential for water purification and remediation of heavy metals in soils; however, its redox reactivity for As(III) sequestration and the corresponding redox-active component are still poorly understood. This study investigated the photochemical properties of GRSP and its mechanism of oxidation/adsorption of As(III). The results showed that UV irradiation triggered electron transfer and the production of reactive oxygen species (ROS) in GRSP, thereby facilitating As(III) oxidation with promotion rates ranging from 43.34 % to 111.1 %. The oxidation of As(III) occurred both on the GRSP photoforming holes and in the ROS reaction from the oxygen reduction products of the photoforming electrons. OH• and H2O2 played an important role in the oxidation of As(III) by GRSP, especially under alkaline conditions. Moreover, the presence of Fe(III) in GRSP facilitated the formation of OH• and its the oxidation capacity towards As(III). The binding of As(III) to the -COOH, -OH, and -FeO groups on the GRSP surface occurred through surface complexation. Overall, these findings provided new insights into the roles of the redox-active moieties and Fe(III) on GRSP in the promoted oxidation of As(III), which would help to deepen our understanding of the migration and transformation of As(III) in soils.

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Processes of water retention and movement and the hydraulic conductivity are altered in the rhizosphere. The aim of this study was to investigate the physical-hydric properties of soil aggregates in the rhizosphere of annual ryegrass (Lolium multiflorum) cropped in a Kandiudalfic Eutrudox, taking into account aspects related to soil aggregate stability. Soil aggregates from rhizosphere soil (RZS) and soil between plant rows (SBP) were used to determine soil water retention curves (SWRCs) and saturated hydraulic conductivity (Ksat). In addition, properties related to soil aggregate stability, such as water-dispersible clay, soil organic carbon (SOC), and microbial activity, were also assessed. The higher microbial activity observed in the RZS was facilitated by increased SOC and microbial activity, resulting in improved soil aggregation (less water-dispersible clay). For nearly all measured matric potentials, RZS had a higher water content than SBP. This was attributed to the stability of aggregates, increase in SOC content, and the root exudates, which improved soil water retention. The increase in total porosity in RZS was associated with improved soil aggregation, which prevents deterioration of the soil pore space and results in higher Ksat and hydraulic conductivity as a function of the effective relative saturation in RZS compared to SBP.

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Corn (Zea mays L.) is an important annual grain that is cultivated as a food staple around the world. The current study examined the effect of wastewater and a combination of biological and organic fertilizers on the morphological and phytochemical traits of corn, using a factorial experiment based on a randomized complete block design with three replications. The first factor was biological and organic fertilizers at seven levels, including the control (no fertilization), bacterial biological fertilizers (NPK) along with iron and zinc Barvar biofertilizers, fungal biofertilizers made from Mycorrhiza and Trichoderma, biochar, a combination of bacterial and fungal biofertilizers, and a combination of bacterial and fungal biofertilizers with biochar. The second factor was irrigation at two levels (conventional irrigation and irrigation with wastewater). The traits studied included the morphological yield, phenols, flavonoids, polyphenols, glomalin, cadmium content in plant parts, and translocation factor (TF). The results disclosed that the best treatment in regard to the morphological traits was related to conventional water + biochar + mycorrhiza + Trichoderma + NPK. The highest phenol and flavonoid content were observed when biochar + mycorrhiza + Trichoderma + NPK treatments were used in both water treatments. Also, the wastewater + biochar + mycorrhiza + Trichoderma + NPK treatment demonstrated the highest total glomalin and phenylalanine ammonia-lyase (PAL) activity. The obtained results demonstrate that combined biological and organic fertilizer use on corn plants can effectively alleviate the deleterious effects of cadmium present in wastewater.

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Although the formation, turnover, and accumulation of soil organic carbon (SOC) are driven by different fertilizer inputs and their subsequent microbial-mediated transformation, the relationship between changes in plant-derived and microbial-derived components and soil microbial life history strategies under different fertilization regimes has not been well explored. In this study, the changes in microbial necromass carbon (MNC), lignin phenols, and glomalin-related soil protein (GRSP), as well as soil microbial life history strategy were determined in a 16-year field experiment in response to different fertilization regimes, including a no-fertilizer control (C), conventional chemical NPK fertilization (NPK), and partial substitutions of the NPK in chemical fertilizers with a low (30 %) or high (60 %) level of straw (0.3S and 0.6S) or cattle manure (0.3M and 0.6M). The results showed that total lignin phenol content and its contribution to SOC were significantly increased by 88.7 % and 74.2 %, respectively, in high-level straw substitution treatment as compared to chemical fertilization. Both high-level straw and cattle manure substitution increased MNC and total GRSP contents, but did not alter their contributions to SOC compared to chemical fertilization. In fertilized treatments, the high-level cattle manure substitution had the lowest and highest bacterial and fungal K/r ratio, respectively. Bacterial K/r ratio was an important factor in predicting bacterial necromass carbon content and there was a significant negative correlation between them. The ratio of ectomycorrhizal to saprotrophic fungi and fungal diversity were important factors for predicting lignin phenol and GRSP contents, respectively. In addition, the SEMs modeling indicated that straw substitution directly affected lignin phenol and MNC accumulation, whereas cattle manure substitution indirectly affected MNC accumulation by affecting microbial life history strategies. In conclusions, agricultural residues inputs support the formation of a multiple carbon pool of SOC compared to chemical fertilization; and microbial life history strategy is an important driver of SOC formation and affects SOC accumulation and stability in agroecosystems.

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Industrial activities pose a significant ecological risk to water resources as they pollute surrounding waters with vanadium (V). Although the contribution of plants and substrates to V removal in constructed wetlands (CWs) has been reported, the role of arbuscular mycorrhizal fungi (AMF) is unclear. The aim of the present study was to investigate the role of AMF in V removal in CWs and to elucidate the underlying mechanisms. Reed plants (Phragmites australis) were inoculated with an AMF strain (Rhizophagus irregularis) in CW columns, creating AMF-inoculated (+AMF) and non-inoculated (-AMF) treatments. Three levels of influent V concentrations (low: 0.50 mg L-1, medium: 1.14 mg L-1 and high: 1.52 mg L-1) were examined. The + AMF treatment showed higher V removal (60%-98%) than the control (40%-82%) in all three conditions, although the difference was not significant in some cases. The mean mycorrhizal effects were 75%, 19%, and 28% for low, moderate, and high influent V concentrations, respectively. The +AMF treatment showed a higher GRSP-bonded V concentration (5.5 mg g-1) than the -AMF treatment (4.0 mg g-1). Furthermore, +AMF treatment showed larger plants with higher V concentrations in their tissues, accompanied by increased biological concentration factors and biological accumulation factors. Given the remarkable positive effect of AMF on V removal, our study suggests that treating AMF in CWs is a worthwhile approach.

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Glomalin-related soil protein (GRSP) is a significant component in the sequestration of heavy metal in soils, but its mechanisms for metal adsorption are poorly known. This study combined spectroscopic data with molecular docking simulations to reveal metal adsorption onto GRSP's surface functional groups at the molecular level. The EXAFS combined with FTIR and XPS analyses indicated that the adsorption of Cd(II), Sr(II), and Ni(II) by GRSP occurred mainly through the coordination of -OH and -COOH groups with the metal. The -COOH and -OH groups bound to the metal as electron donors and the electron density of the oxygen atom decreased, suggesting that electrostatic attraction might be involved in the adsorption process. Two-dimensional correlation spectroscopy revealed that preferential adsorption occurred on GRSP for the metal in sequential order of -COOH groups followed by -OH groups. The presence of the Ni-C shell in the Ni EXAFS spectrum suggested that Ni formed organometallic complexes with the GRSP surface. However, Sr-C and Cd-C were absent in the second shell of the Sr and Cd spectra, which was attributed to the adsorption of Sr and Cd ions with large hydration ion radius by GRSP to form outer-sphere complexes. Through molecular docking simulations, negatively charged residues such as ASP151 and ASP472 in GRSP were found to provide electrostatic attraction and ligand combination for the metal adsorption, which was consistent with the spectroscopic analyses. Overall, these findings provided new insights into the interaction mechanisms between GRSP and metals, which will help deepen our understanding of the ecological functions of GRSP in metal sequestration.

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Afforestation on degraded croplands has been proposed as an effective measure to promote ecosystem functions including soil organic carbon (SOC) sequestration. Glomalin-related soil protein (GRSP) plays a crucial role in promoting the accumulation and stability of SOC. Nevertheless, mechanisms underlying the effects of afforestation on GRSP accumulation have not been well elucidated. In the present study, 14 pairs of maize fields and plantation forests were selected using a paired-site approach in a karst region of southwest China. By measuring soil GRSP and a variety of soil biotic and abiotic variables, the pattern of and controls on GRSP accumulation in response to afforestation were explored. The average content of total GRSP (T-GRSP) and its contribution to SOC in the maize field were 5.22 ± 0.29 mg g-1 and 42.33 ± 2.25%, and those in the plantation forest were 6.59 ± 0.32 mg g-1 and 25.77 ± 1.17%, respectively. T-GRSP content was increased by 26.4% on average, but its contribution to SOC was decreased by 39.1% following afforestation. T-GRSP content decreased as soil depth increased regardless of afforestation or not. Afforestation increased T-GRSP indirectly via its positive effects on arbuscular mycorrhizal fungi biomass, which was stimulated by afforestation through elevating fine root biomass or increasing the availability of labile C and N. The suppressed contribution of T-GRSP to SOC following afforestation was due to the relatively higher increase in other SOC components than T-GRSP and the significant increase of soil C:N ratio. Our study reveals the mechanisms underlying the effects of afforestation on T-GRSP accumulation, and is conducive to improving the mechanistic understanding of microbial control on SOC sequestration following afforestation.

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Glomalin-related soil protein (GRSP) is a stable iron-organic carbon mixture that can enhance heavy metal sequestration in soils. However, the roles of GRSP in the transformation and fate of Cr(VI) have been rarely reported. Herein, we investigated the electrochemical and photocatalytic properties of GRSP and its mechanisms in Cr(VI) adsorption and reduction. Results showed that GRSP had a stronger ability for Cr(VI) adsorption and reduction than other biomaterials, with the highest adsorption amount of up to 0.126 mmol/g. The removal efficiency of Cr(VI) by GRSP was enhanced (4-7%) by ultraviolet irradiation due to the hydrated electrons produced by GRSP. Fe(II) ions, persistent free radicals, and oxygen-containing functional groups on the GRSP surface as electron donors participated in the reduction of Cr(VI) under dark condition. Moreover, Cr(III) was mainly adsorbed on the -COOH groups of GRSP via electrostatic interactions. Based on 2D correlation spectroscopy, the preferential adsorption occurred on the GRSP surface for Cr(VI) in the sequential order of CO → COO- → O-H → C-O. This work provides new insights into the Cr(VI) adsorption and reduction mechanism by GRSP. Overall, GRSP can serve as a natural iron-organic carbon for the photo-reduction of Cr(VI) pollution in environments.

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Coastal wetlands contribute to the mitigation of climate change through the sequestration of "blue carbon". Microbial necromass, lignin, and glycoproteins (i.e., glomalin-related soil proteins (GRSP)), as important components of soil organic carbon (SOC), are sensitive to environmental change. However, their contributions to blue carbon formation and the underlying factors remain largely unresolved. To address this paucity of knowledge, we investigated their contributions to blue carbon formation along a salinity gradient in coastal marshes. Our results revealed decreasing contributions of microbial necromass and lignin to blue carbon as the salinity increased, while GRSP showed an opposite trend. Using random forest models, we showed that their contributions to SOC were dependent on microbial biomass and resource stoichiometry. In N-limited saline soils, contributions of microbial necromass to SOC decreased due to increased N-acquisition enzyme activity. Decreases in lignin contributions were linked to reduced mineral protection offered by short-range-ordered Fe (FeSRO). Partial least-squares path modeling (PLS-PM) further indicated that GRSP could increase microbial necromass and lignin formation by enhancing mineral protection. Our findings have implications for improving the accumulation of refractory and mineral-bound organic matter in coastal wetlands, considering the current scenario of heightened nutrient discharge and sea-level rise.

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Soil pollution caused by organic pollutants and potentially toxic elements poses a serious threat to sustainable agricultural development, global food security and human health. Therefore, strategies for reducing soil pollution are urgently required. Arbuscular mycorrhizal fungi (AMF)-assisted phytoremediation is widely recognized for its ability to remediate slightly-contaminated soil. Glomalin-related soil protein (GRSP) production by AMF is considered a vital mechanism of AMF-assisted phytoremediation. GRSP is widespread in soils and may contribute to the remediation of slightly contaminated soils. GRSP facilitates stabilization of pollutants in soils by interacting with pollutants owing to its abundant functional groups, recalcitrance, and long turnover time. It also enhances soil bioremediation and phytoremediation by stimulating soil microbial activity, improving soil structure, and providing nutrients for plants. However, research on GRSP is still in its early stages, and studies on contaminated soil remediation are limited. The effectiveness of GRSP in situ remediation remains to be proved. This review summarizes current knowledge regarding the GRSP distribution and its contribution to the remediation of slightly contaminated soils. Additionally, we present strategies to increase the GRSP content in contaminated soils, as well as prospects for future studies on the use of GRSP in contaminated soil remediation. This study focuses on recent developments that aim to improve awareness of the role of GRSP in soil remediation and relevant future directions.

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Invasive alien plant species (IAPS) have the ability to change the biochemical properties and the arbuscular mycorrhizal fungal (AMF) community structure in their rhizosphere. Organic acids, microbial activity, and AMF play a key role in the invader's spread and also has interactions with the soil chemical factors. Our aim here was to assess the rhizosphere's biochemical factors, AMF community composition, and soil chemical properties associated with Cryptostegia madagascariensis (IAPS) and Mimosa tenuiflora (endemic plant species) from the Brazilian Seasonal Dry Forest. The highest values of total glomalin (5.87 mg g-1 soil), root colonization (54.5%), oxalic and malic acids (84.21 and 3.01 µmol g-1 , respectively), microbial biomass C (mg kg-1 ), Na+ (0.080 cmolc kg-1 ), Ca2+ (7.04 cmolc kg-1 ), and soil organic carbon (4.59 g kg-1 ) were found in the rhizosphere of C. madagascariensis. We found dissimilarities on AMF community structure considering the studied plant species: (i) Racocetra coralloidea, Dentiscutata heterogama, Dentiscutata cerradensis, Gigaspora decipiens, and AMF's richness were highly correlated with the rhizosphere of M. tenuiflora; and (ii). The rhizosphere of C. madagascariensis was highly correlated with the abundance of Claroideoglomus etunicatum, Rhizoglomus aggregatum, Funneliformis mosseae, and Funneliformis geosporum. The results of our study highlight the importance of considering C. madagascariensis as potential hosts for AMF species from Glomerales, and a potential plant species that increase the bioavailability of exchangeable Na and Ca at semi-arid conditions.

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Rhizophagus irregularis is the model species for arbuscular mycorrhizal fungi (AMF) research and the most widely propagated species for commercial plant biostimulants. Using asymbiotic and symbiotic cultivation systems initiated from single spores, advanced microscopy, Sanger sequencing of the glomalin gene, and PacBio sequencing of the partial 45S rRNA gene, we show that four strains of R. irregularis produce spores of two distinct morphotypes, one corresponding to the morphotype described in the R. irregularis protologue and the other having the phenotype of R. fasciculatus. The two spore morphs are easily distinguished by spore colour, thickness of the subtending hypha, thickness of the second wall layer, lamination of the innermost layer, and the dextrinoid reaction of the two outer spore wall layers to Melzer's reagent. The glomalin gene of the two spore morphs is identical and that of the PacBio sequences of the partial SSU-ITS-LSU region (2780 bp) obtained from single spores of the R. cf fasciculatus morphotype has a median pairwise similarity of 99.8% (SD = 0.005%) to the rDNA ribotypes of R. irregularis DAOM 197198. Based on these results, we conclude that the model AMF species R. irregularis is dimorphic, which has caused taxonomic confusion in culture collections and possibly in AMF research.

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This article focuses on the agronomic evaluation of the supplementation of mineral NPKMg fertilizers with elemental sulfur, magnesium, potassium, or ammonium sulfates in pot experiments with ryegrass growing in a sandy Arenosol with very low sulfur content. A benefit evaluation was carried out on the basis of biomass production, crop nutritional status, and changes in the content of soil organic matter fractions. Furthermore, the total number of bacteria, nitrogen-fixing bacteria, and fungi was estimated using the qPCR technique in soil samples after 60 days of ryegrass growth. The combined application of NPKMg and sulfur or sulfate fertilizers significantly increased the summary yield of three cuttings of fresh ryegrass biomass in the range of 32.3% to 82.7%. The application, especially in the form of sulfates, significantly decreased the content of free phenolic acids. Furthermore, compared to the control, i.e., soil with NPKMg applied alone, an increase in the content of glomalin-related proteins and a decrease in the amount of water-soluble organic carbon compounds were observed. Neither the number of DNA marker copy numbers of the total bacterial community nor the nitrogen-fixing bacteria were noticeably different. In turn, the total number of genetic markers for fungi was significantly higher in soils with potassium or ammonium sulfates compared to the control soil. The general results suggest that the application of sulfur fertilizers with NPKMg mineral fertilizer can benefit crops and support soil fertility due to the stabilization of aggregates and the decrease in water-soluble organic compounds.

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Glomalin-related soil protein (GRSP) has gained widespread attention because of its benefits to carbon sequestration, improving soil quality and fixing heavy metals. However, studies on how GRSP affects the environmental fate of organic contaminants are scarce. In this study, different types of GRSPs were isolated from forest soils and characterized to study the binding of GRSPs and PAHs under different environmental conditions. The results indicated that GRSPs contain abundant functional groups (such as -NH, -COOH, and CO) and material composition, like humic acid, proteins, and lipids. For the tested GRSPs, EE-GRSP has lower DOC, SUVA260 and SUVA280 values, as well as higher E2/E3 values, indicating that EE-GRSP has lower hydrophobicity and molecular weight. These properties can lead to strong interactions between GRSP and PAHs, especially with benzopyrene, which has a high Kow and Ksw and a large molecular size, with binding constants ranging from 16,119 to 163,697 L·kg-1. Furthermore, low pH (pH = 3) and temperature (15 °C) could increase GRSP's aggregation, enhance the GRSP binding ability with PAHs, whose binding constants were 11,595 and 5067.3 L·kg-1. Therefore, the binding between GRSP and PAHs may lead to changes in the fate of PAHs in the soil and affect the environmental risk of PAHs. The results presented here will deepen our understanding of the environmental function of GRSPs and provide a theoretical basis to further elucidate the mechanisms of GRSPs and organic pollutants.

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Glomalin-related soil protein (GRSP) is a hydrophobic protein released by arbuscular mycorrhizal fungi. It is an important component of the soil carbon pool, and it improves the soil aggregate structure; however, it remains unclear whether GRSP can enhance soil carbon sequestration and improve soil quality during rapid urbanization. The built-up area in Nanchang, China was the study area, and the proportion of impervious surface area was the parameter of urbanization intensity. A total of 184 plots (400 m2) were set up to collect soil samples (0-20 cm) for analysis. Aggregates of five particle sizes were sieved, and the percentage amounts of soil organic carbon (SOC) and GRSP for them were determined. The results showed that the easily extractable GRSP (EE-GRSP) and total GRSP (T-GRSP) contents of the four aggregates of <2 mm were 22-46% higher in low urbanization areas than those in high urbanization areas (p < 0.05), indicating that the higher urbanization intensity was associated with the lower GRSP content of different aggregates. The GRSP was significantly positively correlated with SOC (p < 0.05). Moreover, the contribution of GRSP to the SOC pool in the <0.25 mm aggregate was significantly higher than that in other aggregates. In addition, the EE-GRSP content was significantly positively correlated with mean weight diameter (MWD) and geometric mean diameter (GMD) in the four aggregates of <2 mm, whereas it was negatively correlated with fractal dimension (D) in the >2 mm, 1-2 mm and <0.053 mm aggregates. The T-GRSP content showed significant correlations only with MWD, GMD, and D in the 1-2 mm aggregate. This study revealed that increasing urbanization intensity can significantly reduce the GRSP content of different sized aggregates. Moreover, the GRSP content significantly promoted SOC sequestration, and the EE-GRSP content more significantly promoted soil aggregate stability than that of the T-GRSP. These findings provide new ideas for exploring the improvement of soil quality during the process of urbanization.

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Acid rain is one of most serious environment problems in China. The types of acid rain have gradually transformed from sulfuric acid rain (SAR) to mixed acid rain (MAR) and nitric acid rain (NAR) in recent years. Roots are one source of soil organic carbon and play an important role in the formation of soil aggregates. However, the changes in acid rain type and the effect of root removal on soil organic carbon in forest ecosystems are poorly understood. In this study, we removed roots and simulated acid rain with different types (SO42-:NO3- ratio of 4:1, 1:1, and 1:4) for three years in Cunninghamia lanceolata (CP) and Michelia macclurei (MP) plantations to analyze the changes of soil organic carbon and physical properties and to measure the size and mean weight diameter (MWD) of aggregates. Results showed that root removal in C. lanceolata and M. macclurei remarkably reduced soil organic carbon pool by 16.7 % and 21.5%, and soil recalcitrant carbon by 13.5% and 20.0%, respectively. Root removal substantially decreased the MWD and proportion and organic carbon content of soil macroaggregates in M. macclurei, but not in C. lanceolata. Acid rain did not affect soil organic carbon pool and soil aggregate structures. Our results indicated that roots promote soil organic carbon stabilization and that their contribution to the stability of soil organic carbon varies with forest types. Moreover, soil organic carbon stabilization is not affected by different types of acid rain in the short term.

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Glomalin-related soil protein (GRSP) is a widespread recalcitrant soil protein complex that promotes the immobilization of metals in soils. Herein, we combined indoor simulation and field investigation to reveal the interfacial behaviors and ecological functions of GRSP to the three typical metals (Sr(II), Ni(II), and Cd(II)). The kinetic and isotherm data suggested that GRSP had a strong ability to adsorb the metals, which was closely related to the Hard-Soft-Acid-Base theory and the film diffusion mechanisms. Regarding environmental factors, the higher solution pH was beneficial to the adsorption of the metals onto GRSP, while the adsorption capacity decreased at lower or higher salinity due to the salting-out and Na+ competition effects. Moreover, Sr(II), Ni(II), and Cd(II) showed competitive adsorption onto GRSP, which was associated with the spatial site resistance effect. By comparing the retention factors of seven natural and artificial particles, GRSP had elevated distribution coefficients in high metal concentration, while its retention factors showed a relatively lower decrease, suggesting that GRSP had excellent buffer performance for a potential metal pollution emergency. Through the continental-scale coastal regions investigation, GRSP sequestered 1.05-3.11 µmol/g Ni, 0.31-1.49 µmol/g Sr, and 0.01-0.06 µmol/g Cd with 0.54-0.91 % of the sediment mass, demonstrating its strong ability to adsorb the metals. Therefore, we advocate that GRSP, as a recalcitrant protein complex, can be considered an effective tool for buffering capacity of metal pollution and environmental capacity within coastal wetlands.

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Agricultural ecosystems contribute significantly to atmospheric emissions of soil nitrous oxide (N2O), which exacerbate environmental pollution and contribute to global warming. Glomalin-related soil protein (GRSP) stabilizes soil aggregates and enhances soil carbon and nitrogen storage in agricultural ecosystems. However, the underlying mechanisms and relative importance of GRSP on N2O fluxes within soil aggregate fraction remain largely unclear. We examined the GRSP content, denitrifying bacterial community composition, and potential N2O fluxes across three aggregate-size fractions (2000-250 µm, 250-53 µm, and <53 µm) under a long-term fertilization agricultural ecosystem, subjected to mineral fertilizer or manure and their combination. Our findings indicated that various fertilization treatments have no discernible impact on the size distribution of soil aggregates, paving the way to further research into the impact of soil aggregates on GRSP content, the denitrifying bacterial community composition, and potential N2O fluxes. GRSP content increased with the increase in soil aggregate size. Potential N2O fluxes (including gross N2O production and N2O reduction and net N2O production) among aggregates were highest in microaggregates (250-53 µm), followed by macroaggregates (2000-250 µm) and lowest in silt + clay (<53 µm) fractions. Potential N2O fluxes had a positive response to soil aggregate GRSP fractions. The non-metric multidimensional scaling analysis revealed that soil aggregate size could drive the denitrifying functional microbial community composition, and deterministic processes play more critical roles than stochasticity processes in driving denitrifying functional composition under soil aggregate fractions. Procrustes analysis revealed a significant correlation between denitrifying microbial community, soil aggregate GRSP fractions, and potential N2O fluxes. Our study suggests that soil aggregate GRSP fractions influence potential nitrous oxide fluxes by affecting denitrifying microbial functional composition within soil aggregate.

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The relationship between glomalin-related soil protein (GRSP) and soil aggregation has been a hot topic of research for its close link to soil stability and quality. However, the short-term cultivation of Eucalyptus poses serious threats to soil stability and nutrient stocks, and the effects of GRSP on soil aggregate stability and macronutrient accumulation remain unclear. The aim is to clarify the potential mechanisms affecting soil aggregate stability and macronutrient accumulation in short-term Eucalyptus plantations. Five Eucalyptus urophylla × Eucalyptus grandis plantations with different cultivation periods (1-5 years) in this study were investigated, and a native evergreen broadleaf forest (0 year) was selected as control. The mean weight diameter index increased in the first 3 years and then significantly decreased during 5 years cultivation of Eucalyptus. Soil organic carbon (SOC) and total nitrogen also decreased after planting Eucalyptus for 3 years, but variation in total phosphorus was not obvious. The relative abundance of Glomeraceae and Claroideoglomeraceae decreased in the 5-year-old Eucalyptus plantations and was positively correlated with GRSP content. In pathway modeling, nutrient-acquisition enzyme activities positively affected GRSP and macronutrient content. Total GRSP (T-GRSP) had higher total effects than easily extractable GRSP on soil aggregate stability, and positively correlated with SOC in macroaggregates. Both T-GRSP and SOC had positive and direct effects on soil aggregate stability. Variance partitioning analysis further explained the contribution of GRSP and SOC to aggregate stability, particularly in >2 and 2-0.25 mm macroaggregates. Our results suggested that GRSP was directly associated with SOC content and soil aggregate stability, and was a potential key factor affecting soil aggregate stability in Eucalyptus plantations. Improving T-GRSP and SOC are efficient approaches for preventing the gradual deterioration of soil aggregate stability. Short-term cultivation should be carefully used in Eucalyptus plantations, and a new cultivation period is needed.

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Technogenic soil (technosol) developed from coal fly ash (FA) landfilling has been considered a critical environmental problem worldwide. Drought-tolerant plants often naturally grow on FA technosol. However, the impact of these natural revegetations on the recovery of multiple ecosystem functions (multifunctionality) remains largely unexplored and poorly understood. Here we assessed the response of multifunctionality, including nutrient cycling (i.e., carbon, nitrogen, and phosphorus), carbon storage, glomalin-related soil protein (GRSP), plant productivity, microbial biomass carbon (MBC), microbial processes (soil enzyme activities), and soil chemical properties (pH and electrical conductivity; EC) to FA technosol ten years' natural revegetation with different multipurpose species in Indo-Gangetic plain, and identified the key factors regulating ecosystem multifunctionality during reclamation. We evaluated four dominant revegetated species: Prosopis juliflora, Saccharum spontaneum, Ipomoea carnea, and Cynodon dactylon. We found that natural revegetation initiated the recovery of ecosystem multifunctionality on technosol, with greater recovery under higher biomass-producing species (P. juliflora and S. spontaneum) than lower biomass-producing ones (I. carnea and C. dactylon). The individual functions (11 of the total 16 variables) at higher functionality (70 % threshold) also exhibited this pattern among revegetated stands. Multivariate analyses revealed that most of the variables (except EC) significantly correlated with multifunctionality, indicating the capability of multifunctionality to consider the tradeoff between individual functions. We further performed structural equation modeling (SEM) to detect the effect of vegetation, pH, nutrients, and microbial activity (MBC and microbial processes) on ecosystem multifunctionality. Our SEM model predicted 98 % of the variation in multifunctionality and confirmed that the indirect effect of vegetation mediated by microbial activity is more important for multifunctionality than their direct effect. Collectively, our results demonstrate that FA technosol revegetation with high biomass-producing multipurpose species promotes ecosystem multifunctionality and emphasizes the significance of microbial activity in the recovery and maintenance of ecosystem attributes.

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