Arbuscular mycorrhizal fungal interactions bridge the support of root-associated microbiota for slope multifunctionality in an erosion-prone ecosystem.

The role of diverse soil microbiota in restoring erosion-induced degraded lands is well recognized. Yet, the facilitative interactions among symbiotic arbuscular mycorrhizal (AM) fungi, rhizobia, and heterotrophic bacteria, which underpin multiple functions in eroded ecosystems, remain unclear. Here, we utilized quantitative microbiota profiling and ecological network analyses to explore the interplay between the diversity and biotic associations of root-associated microbiota and multifunctionality across an eroded slope of a Robinia pseudoacacia plantation on the Loess Plateau. We found explicit variations in slope multifunctionality across different slope positions, associated with shifts in limiting resources, including soil phosphorus (P) and moisture. To cope with P limitation, AM fungi were recruited by R. pseudoacacia, assuming pivotal roles as keystones and connectors within cross-kingdom networks. Furthermore, AM fungi facilitated the assembly and composition of bacterial and rhizobial communities, collectively driving slope multifunctionality. The symbiotic association among R. pseudoacacia, AM fungi, and rhizobia promoted slope multifunctionality through enhanced decomposition of recalcitrant compounds, improved P mineralization potential, and optimized microbial metabolism. Overall, our findings highlight the crucial role of AM fungal-centered microbiota associated with R. pseudoacacia in functional delivery within eroded landscapes, providing valuable insights for the sustainable restoration of degraded ecosystems in erosion-prone regions.

Nutrient limitation mediates soil microbial community structure and stability in forest restoration.

Soil microorganisms are often limited by nutrients, representing an important control of heterotrophic metabolic processes. However, how nutrient limitations relate to microbial community structure and stability remains unclear, which creates a knowledge gap to understanding microbial biogeography and community changes during forest restoration. Here, we combined an eco-enzymatic stoichiometry model and high-throughput DNA sequencing to assess the potential roles of nutrient limitation on microbial community structure, assembly, and stability along a forest restoration sequence in the Qinling Mountains, China. Results showed that nutrient limitations tended to decrease during the oak forest restoration. Carbon and phosphorus limitations enhanced community dissimilarity and significantly increased bacterial alpha diversity, but not fungal diversity. Stochastic assembly processes primarily structured both bacterial (average contribution of 74.73 % and 74.17 % in bulk and rhizosheath soils, respectively) and fungal (average contribution of 77.23 % and 72.04 % in bulk and rhizosheath soils, respectively) communities during forest restoration, with nutrient limitation also contributing to the importance of stochastic processes in the bacterial communities. The migration rate (m) for bacteria was 0.19 and 0.23, respectively in both bulk soil and rhizosheath soil, and was greater than that for the fungi (m was 1.19 and 1.41, respectively), indicating a stronger dispersal limitation for fungal communities. Finally, nutrient limitations significantly affected bacterial and fungal co-occurrence with more interconnections occurring among weakly nutrient-limited microbial taxa and nutrient limitations reducing community stability when nutrient availability changed during forest restoration. Our findings highlight the fundamental effects of nutrient limitations on microbial communities and their self-regulation under changing environmental resources.

Deciphering microbiomes dozens of meters under our feet and their edaphoclimatic and spatial drivers.

Microbes inhabiting deep soil layers are known to be different from their counterpart in topsoil yet remain under investigation in terms of their structure, function, and how their diversity is shaped. The microbiome of deep soils (>1 m) is expected to be relatively stable and highly independent from climatic conditions. Much less is known, however, on how these microbial communities vary along climate gradients. Here, we used amplicon sequencing to investigate bacteria, archaea, and fungi along fifteen 18-m depth profiles at 20-50-cm intervals across contrasting aridity conditions in semi-arid forest ecosystems of China's Loess Plateau. Our results showed that bacterial and fungal α diversity and bacterial and archaeal community similarity declined dramatically in topsoil and remained relatively stable in deep soil. Nevertheless, deep soil microbiome still showed the functional potential of N cycling, plant-derived organic matter degradation, resource exchange, and water coordination. The deep soil microbiome had closer taxa-taxa and bacteria-fungi associations and more influence of dispersal limitation than topsoil microbiome. Geographic distance was more influential in deep soil bacteria and archaea than in topsoil. We further showed that aridity was negatively correlated with deep-soil archaeal and fungal richness, archaeal community similarity, relative abundance of plant saprotroph, and bacteria-fungi associations, but increased the relative abundance of aerobic ammonia oxidation, manganese oxidation, and arbuscular mycorrhizal in the deep soils. Root depth, complexity, soil volumetric moisture, and clay play bridging roles in the indirect effects of aridity on microbes in deep soils. Our work indicates that, even microbial communities and nutrient cycling in deep soil are susceptible to changes in water availability, with consequences for understanding the sustainability of dryland ecosystems and the whole-soil in response to aridification. Moreover, we propose that neglecting soil depth may underestimate the role of soil moisture in dryland ecosystems under future climate scenarios.

Deciphering the dual role of bacterial communities in stabilizing rhizosphere priming effect under intra-annual change of growing seasons.

The rhizosphere priming effect (RPE) is a widely observed phenomenon affecting carbon (C) turnover in plant-soil systems. While multiple cropping and seasonal changes can have significant impacts on RPE, the mechanisms driving these processes are complex and not yet fully understood. Here, we planted maize in paddy soil during two growing seasons having substantial temperature differences [May-August (warm season, 26.6 °C) and September-November (cool season, 23.1 °C)] within the same calendar year in southern China to examine how seasonal changes affect RPEs and soil C. We identified sources of C emissions by quantifying the natural abundance of 13C and determined microbial metabolic limitations or efficiency and functional genes related to C cycling using an enzyme-based biogeochemical equilibrium model and high-throughput quantitative PCR-based chip technology, respectively. Results showed that microbial metabolism was mainly limited by phosphorus in the warm season, but by C in the cool season, resulting in positive RPEs in both growing seasons, but no significant differences (9.02 vs. 6.27 mg C kg-1 soil day-1). The RPE intensity remained stable as temperature increased (warm season compared to a cool season), which can be largely explained by the simultaneous increase in the abundance of functional genes related to both C degradation and fixation. Our study highlights the simultaneous response and adaptation of microbial communities to seasonal changes and hence contributes to an understanding and prediction of microbially mediated soil C turnover under multiple cropping systems.

Microbial communities in terrestrial surface soils are not widely limited by carbon.

Microbial communities in soils are generally considered to be limited by carbon (C), which could be a crucial control for basic soil functions and responses of microbial heterotrophic metabolism to climate change. However, global soil microbial C limitation (MCL) has rarely been estimated and is poorly understood. Here, we predicted MCL, defined as limited availability of substrate C relative to nitrogen and/or phosphorus to meet microbial metabolic requirements, based on the thresholds of extracellular enzyme activity across 847 sites (2476 observations) representing global natural ecosystems. Results showed that only about 22% of global sites in terrestrial surface soils show relative C limitation in microbial community. This finding challenges the conventional hypothesis of ubiquitous C limitation for soil microbial metabolism. The limited geographic extent of C limitation in our study was mainly attributed to plant litter, rather than soil organic matter that has been processed by microbes, serving as the dominant C source for microbial acquisition. We also identified a significant latitudinal pattern of predicted MCL with larger C limitation at mid- to high latitudes, whereas this limitation was generally absent in the tropics. Moreover, MCL significantly constrained the rates of soil heterotrophic respiration, suggesting a potentially larger relative increase in respiration at mid- to high latitudes than low latitudes, if climate change increases primary productivity that alleviates MCL at higher latitudes. Our study provides the first global estimates of MCL, advancing our understanding of terrestrial C cycling and microbial metabolic feedback under global climate change.

Crop residue return sustains global soil ecological stoichiometry balance.

Although soil ecological stoichiometry is constrained in natural ecosystems, its responses to anthropogenic perturbations are largely unknown. Inputs of inorganic fertilizer and crop residue are key cropland anthropogenic managements, with potential to alter their soil ecological stoichiometry. We conducted a global synthesis of 682 data pairs to quantify the responses of soil carbon (C), nitrogen (N), and phosphorus (P) and grain yields to combined inputs of crop residue plus inorganic fertilizer compared with only inorganic fertilizer application. Crop residue inputs enhance soil C (10.5%-12%), N (7.63%-9.2%), and P (2.62%-5.13%) contents, with an increase in C:N (2.51%-3.42%) and C:P (7.27%-8.00%) ratios, and grain yields (6.12%-8.64%), indicating that crop residue alleviated soil C limitation caused by inorganic fertilizer inputs alone and was able to sustain balanced stoichiometry. Moreover, the increase in soil C and C:N(P) ratio reached saturation in ~13-16 years after crop residue return, while grain yield increase trend discontinued. Furthermore, we identified that the increased C, N, and P contents and C:N(P) ratios were regulated by the initial pH and C content, and the increase in grain yield was not only related to soil properties, but also negatively related to the amount of inorganic N fertilizer input to a greater extent. Given that crop residual improvement varies with soil properties and N input levels, we propose a predictive model to preliminary evaluate the potential for crop residual improvement. Particularly, we suggest that part of the global budget should be used to subsidize crop residue input management strategies, achieving to a win-win situation for agricultural production, ecological protection, and climate change mitigation.

Microbial nutrient limitations limit carbon sequestration but promote nitrogen and phosphorus cycling: A case study in an agroecosystem with long-term straw return.

Soil fertility can be increased by returning crop residues to fields due to the cooperative regulation of microbial metabolism of carbon (C) and nutrients. However, the dose-effect of straw on the soil C and nutrient retention and its underlying coupled microbial metabolic processes of C and nutrients remain poorly understood. Here, we conducted a comprehensive study on soil nutrients and stoichiometry, crop nutrient uptake and production, microbial metabolic characteristics and functional attributes using a long-term straw input field experiment. We estimated the microbial metabolic limitations and efficiency of C and nitrogen (N) use (CUE and NUE) via an enzyme-based vector-TER model, biogeochemical-equilibrium model and mass balance equation, respectively. In addition, the absolute abundances of 20 functional genes involved in the N- and P-cycles were quantified by quantitative PCR-based chip technology. As expected, straw input significantly increased C and N stocks, C: nutrients, crop nutrient uptake and growth. However, the C sequestration efficiency decreased by approximately 6.1 %, and the N2O emission rate increased by 0.5-1.0 times with the increase in straw input rate. Interestingly, the microbial metabolism was more limited by P when straw input was <8 t ha-1 but was reversed when straw input was 12 t ha-1. The enhanced nutrient limitation reduced both the CUE and the NUE of microbes and then upregulated genes associated with the hydrolysis of C, the mineralization of N and P, and denitrification, which consequently influenced C and N losses as well as crop growth. This study highlights that soil C and nutrient cycling are strongly regulated by microbial metabolic limitation, suggesting that adding the appropriate limiting nutrients to reduce nutrient imbalances caused by straw input is conducive to maximizing the ecological benefits of straw return.

Synergistic interplay between Azospirillum brasilense and exogenous signaling molecule H2S promotes Cd stress resistance and growth in pak choi (Brassica chinensis L.).

Inoculation with growth-promoting rhizobacteria inoculation and the addition of exogenous signaling molecules are two distinct strategies for improving heavy metal resistance and promoting growth in crops through several mechanisms. However, whether rhizobacteria and phyllosphere signaling molecules can act synergistically alleviate heavy metal stress and promote growth and the mechanisms underlying these effects remain unclear. Here, a novel strategy involving the co-application of growth-promoting rhizobacteria and an exogenous signaling molecule was developed to reduce cadmium (Cd) phytotoxicity and promote pak choi growth in Cd-contaminated soil. We found that the co-application of Azospirillum brasilense and hydrogen sulfide (H2S) resulted in significant improvements in shoot biomass and antioxidant enzyme content and a decline in the levels of Cd translocation factors. In addition, this co-application significantly improved pak choi Cd resistance. Furthermore, we observed a significant negative correlation between abscisic acid concentration and Cd content of pak choi and a positive correlation between H2S concentration and biomass. These findings revealed that the co-application of rhizobacteria and exogenous signaling molecules synergistically promoted the growth of vegetable crops subjected to heavy metal stress. Our results may serve as a guide for improving the food safety of crops grown in soil contaminated with heavy metals.

Quantifying Methane Emissions from Aquaculture Ponds in China.

Small ponds are important methane (CH4) sources. However, current estimates of CH4 emissions from aquaculture ponds are largely uncertain due to data paucity, especially in China─the largest aquaculture producer in the world. Here, we present a nationwide metadata analysis with a database of 55 field observations to examine total CH4 emissions from aquaculture ponds in China. We found that the annual CH4 fluxes from aquaculture ponds are much larger than those from reservoirs and lakes. The total CH4 emission from aquaculture ponds is 1.60 ± 0.62 Tg CH4 yr-1, with an average growth rate of ∼0.03 Tg CH4 yr-2 during the period 2008-2019. Compared with global major protein-producing livestocks, aquaculture species have a lower (63%) emission intensity, defined by the amount of CH4 emitted per unit of animal proteins. Our study highlights the essential contribution of China's aquaculture ponds to national CH4 emissions and the lower environmental cost of the aquaculture sector for future animal protein production. More field measurements with multi-scale observations are urgently needed to reduce the uncertainty of CH4 emissions from aquaculture ponds.

Microbial metabolic limitation and carbon use feedback in lead contaminated agricultural soils.

Potentially toxic elements (PTEs) pollution causes a great threat to microbial metabolism, which plays a vital role in studying soil nutrient cycling and predicting carbon (C) storage in agroecosystems. However, the responses of microbial metabolism characteristic to heavy metal contamination and the mechanisms through which microbial metabolism mediate nutrient cycling and C dynamics in contaminated soil remain elusive. Here, we performed an incubation experiment over 80 days to investigate the variations in microbial metabolic limitation under various Pb levels (0, 100, 500, 800, 1500, 2000, and 3000 mg Pb kg-1 dry soil) in cropland soil using extracellular enzymatic stoichiometry, and to reveal the impact of Pb stress on soil C storage by associating with microbial metabolic quotients (qCO2) and C use efficiency (CUE). The results showed microbial relative C limitation and phosphorus (P) limitation were observed in Pb-contaminated soils. Pb addition enhanced the microbial relative C limitation by approximately 7.3%, while decreasing the P limitation by approximately 12.3%. Furthermore, Pb addition led to higher qCO2 (from 8.75 to 108 µg C kg-1 MBC-1 d-1) duo to the increase of microbial relative C limitation, suggesting that the more CO2 was released of per unit of microbial biomass C. The increase of microbial relative C limitation reduced CUE (from 0.35 to 0.10) because of the change in microbial metabolism from growth to respiration maintenance under Pb stress. Consequently, the CUE and qCO2 together determined the loss of soil C. Our study reveals that microbial relative C limitation is the dominant driver of soil C loss and provides important knowledge of microbial metabolic limitation regulating soil C turnover in PTEs-contaminated agricultural soils.

Consistent Plant and Microbe Nutrient Limitation Patterns During Natural Vegetation Restoration.

Vegetation restoration is assumed to enhance carbon (C) sequestration in terrestrial ecosystems, where plant producers and microbial decomposers play key roles in soil C cycling. However, it is not clear how the nutrient limitation patterns of plants and soil microbes might change during vegetation restoration. We investigated the nutrient limitations of the plant and microbial communities along a natural vegetation restoration chronosequence (1, 8, 16, 31, and 50 years) following farmland abandonment in Qinling Mountains, China, and assessed their relationships with soil factors. The result showed that following natural vegetation restoration, the nitrogen (N) limitation of plant and microbial communities was alleviated significantly, and thereafter, it began to shift to phosphorus (P) limitation at a later stage. Plants showed P limitation 50 years after restoration, while microbial P limitation appeared 31 years later. The changes in plant nutrient limitation were consistent with those in microbial nutrient limitation, but soil microbes were limited by P earlier than plants. Random forest model and partial least squares path modeling revealed that soil nutrient stoichiometry, especially soil C:N ratio, explained more variations in plant and microbial nutrient limitation. Our study demonstrates that the imbalanced soil C:N ratio may determine the soil microbial metabolic limitation and further mediate the variation in plant nutrient limitation during natural vegetation restoration, which provides important insights into the link between metabolic limitation for microbes and nutrient limitation for plants during vegetation restoration to improve our understanding of soil C turnover in temperate forest ecosystems.

Microbial metabolic limitation of rhizosphere under heavy metal stress: Evidence from soil ecoenzymatic stoichiometry.

Slow nutrient turnover and destructed soil function were the main factors causing low efficiency in phytoremediation of heavy metal (HM)-contaminated soil. Soil ecoenzymatic stoichiometry can reflect the ability of soil microorganisms to acquire energy and nutrients, and drive nutrient cycling and carbon (C) decomposition in HM-contaminated soil. Therefore, for the first time, we used the enzymatic stoichiometry modeling to examine the microbial nutrient limitation in rhizospheric and bulk soil of different plants (Medicago sativa, Halogeton arachnoideus and Agropyron cristatum) near the Baiyin Copper Mine. Results showed that the main pollutants in this area were Cu, Zn, Cd, and Pb, while Cd and Zn have the greatest contribution according to the analysis of pollution load index (PLI). The activities of soil C-, nitrogen (N)-, and phosphorus (P)-acquiring enzymes in the rhizosphere of plants were significantly greater than that in bulk soil. Moreover, microbial C and P limitations were observed in all plant treatments, while the lower limitation was generally in the rhizosphere compared to bulk soil. The HM stress significantly increased microbial C limitation and decreased microbial P limitation, especially in the rhizospheric soil. The partial least squares path modeling (PLS-PM) further indicated that HM concentration has the greatest effects on microbial P limitation (-0.64). In addition, the highest enzyme activities and the lowest P limitation were observed in the rhizospheric and bulk soil of M. sativa, thereby implying that soil microbial communities under the remediation of M. sativa were steadier and more efficient in terms of their metabolism. These findings are important for the elucidation of the nutrient cycling and microbial metabolism of rhizosphere under phytoremediation, and provide guidance for the restoration of HM-contaminated soil.

The impacts of nitrogen addition on upland soil methane uptake: A global meta-analysis.

Elevated nitrogen (N) addition from anthropogenic activities has great impacts on soil methane (CH4) uptake, which could interrupt the existing global CH4 balance and cause feedbacks to climate and biogeochemical processes. Previous studies have come to inconsistent conclusions on both the quantification of the response of CH4 uptake to N addition and understanding of its underlying mechanisms, probably due to the lack of experimental data. Here, we conduct a broad meta-analysis of 90 papers to quantify the responses of CH4 uptake to N addition in upland soil. The results show that N addition has a significant negative impact on soil CH4 uptake (-19.25%), which is termed the N inhibition effect. Soil pH is identified as the dominant factor, with the other factors affecting the CH4 uptake through the alteration of soil pH. The N inhibition effect is observed to be large and significant in forest and grassland, but small and insignificant in farmland, because of the distinct composition of their methanotrophic communities. A threshold of the N addition level is identified at about 68 kg N ha-1 year-1, which indicates the lowest N inhibition effect. Furthermore, the convex relationship between response ratio of CH4 uptake (negative) and N addition duration indicates that a medium level of N addition duration has the largest N inhibition effect, and longer or shorter durations will both reduce the effect. Our analysis of the N inhibition effect implies that controlling the N addition level could effectively reduce the CH4 concentration in the atmosphere and thus relieve global warming.

Improvement of alfalfa resistance against Cd stress through rhizobia and arbuscular mycorrhiza fungi co-inoculation in Cd-contaminated soil.

Rhizobia and arbuscular mycorrhiza fungi (AMF) are important symbiotic microbes that are advantageous to plants growing in metal-contaminated soil. However, it remains unclear how inoculated microbes affect rhizosphere microbial communities or whether subsequent changes in rhizosphere microbiomes contribute to improving plant resistance under metal stress. This study investigated the effects of rhizobia and AMF inoculation on alfalfa resistance to Cd stress. The response of rhizosphere microbial communities to inoculation and its role in increasing alfalfa' ability to cope with stress were further analyzed using high-throughput sequencing of 16S and ITS rRNA genes. Results showed that single rhizobia or AMF inoculation significantly improved alfalfa resistance to Cd stress, while their co-inoculation resulted in the greatest overall improvement. Improved resistance was reflected by the significant mitigation of Cd-induced lipid peroxidation and reactive oxygen species (ROS) stress caused by increases in antioxidant enzyme activities along with co-inoculation. Furthermore, co-inoculation significantly altered the rhizosphere microbial community structure by decreasing fungal community diversity and increasing bacterial community diversity. Results of partial least squares path modeling (PLS-PM) and variation partitioning analysis (VPA) showed that the rhizosphere bacterial community predominated over the fungal community with respected to improvements in resistance to Cd stress under the co-inoculation treatments. This improvement was specifically seen in the enrichment of certain key bacterial taxa (including Proteobacteria, Actinobacteria, Acidobacteria, and Chloroflexi) induced by the rhizobia and AMF co-inoculation, enhancing alfalfa' ability to uptake rhizosphere nutrients and reduce its release of photosynthetically-derived carbon (C) into soil. Our findings revealed that the co-inoculation of multiple symbiotic microbes can assist plants to effectively cope with Cd stress, providing a greater understanding of rhizosphere bacterial taxa in the microbe-induced phytomanagement.

A novel extracellular enzyme stoichiometry method to evaluate soil heavy metal contamination: Evidence derived from microbial metabolic limitation.

Heavy metal contaminates have become a significant threat to soil ecosystems due to their chronicity and universality in soil. Soil microbial metabolism plays a vital role in biogeochemical cycles and soil functions. However, the response of microbial metabolism to heavy metal contamination in soil remains elusive despite potentially offering important insight into the health and ecological consequences of soil ecosystems under such contamination. This study used extracellular enzyme stoichiometry models to identify the response of microbial metabolism to various heavy metal contaminants, while also revealing potential implications of heavy metal contaminates in soil ecosystems. Results showed that microbial metabolism was restricted by soil carbon (C) and phosphorus (P) within a heavy metal polluted area in Northwest China. Heavy metal stress significantly increased microbial C limitation while decreasing microbial P limitation. However, microbial C and P limitations both responded consistently to different heavy metals (i.e., Cd, Pb, Zn, and Cu). Heavy metals had the greatest effect on microbial C limitation (i.e., 0.720 of the total effects) compared to other soil properties, and soil with the lowest heavy metal concentration exhibited the lowest microbial C limitation, and vice versa. These results indicated that microbial metabolic limitation can robustly and sensitively reflect the degree of heavy metals pollution in soil. Additionally, increased microbial C limitation caused by heavy metal contaminants could potentially escalate C release by promoting soil C decomposition as well as increasing investments in enzyme production and the maintenance of metabolic processes. Consequently, potential C loss induced by heavy metal pollution on soil ecosystems may be extensive and significant. Generally, our results suggest the usefulness of extracellular enzyme stoichiometry as a new method from which to evaluate heavy metal soil pollution, while microbial metabolic limitation could potentially be a promising indicator.

Phosphorus recovery by core-shell γ-Al2O3/Fe3O4 biochar composite from aqueous phosphate solutions.

Biochar can act as an adsorbent for phosphate removal from water sources, which can be highly beneficial in limiting eutrophication and recycling elemental phosphorus (P). However, it is difficult to use a single biochar material to overcome problems such as low adsorption efficiency, difficulty in reuse, and secondary pollution. This study addresses these challenges using a novel core-shell structure γ-Al2O3/Fe3O4 biochar adsorbent (AFBC) with significant P uptake capabilities in terms of its high adsorption capacity (205.7 mg g-1), magnetic properties (saturation magnetization 24.70 emu g-1), and high reuse stability (91.0% removal efficiency after five adsorption-desorption cycles). The highest partition coefficient 1.04 mg g-1 µM-1, was obtained at a concentration of 322.89 µM. Furthermore, AFBC exhibited strong regeneration ability in multiple cycle trials, making it extremely viable for sustainable resource management. P removal mechanisms, i.e., electrostatic attraction and inner-sphere complexation, were explained using Fourier transform infrared (FT-IR) spectra and X-ray photoelectron spectroscopy (XPS) measurements. A surface complexation model was established by considering the formation of monodentate mononuclear and bidentate binuclear surface complexes of P to illustrate the adsorption process. Owing to its high adsorption efficiency, easy separation from water, and environmental friendliness, AFBC is a potential adsorbent for P recovery from polluted waters.

Co-inoculation effect of plant-growth-promoting rhizobacteria and rhizobium on EDDS assisted phytoremediation of Cu contaminated soils.

Chelants application can increase the bioavailability of metals, subsequently limiting plant growth and reducing the efficiency of phytoremediation. Plant growth-promoting rhizobacteria (PGPRs) and rhizobium have substantial potential to improve plant growth and plant tolerance to metal stress. We evaluated the effects of co-inoculation with a PGPR strain (Paenibacillus mucilaginosus) and a Cu-resistant rhizobium strain (Sinorhizobium meliloti) on the efficiency of biodegradable chelant (S,S-ethylenediaminedisuccinic acid; EDDS) assisted phytoremediation of a Cu contaminated soil using alfalfa. The highest total Cu extraction by alfalfa was observed in the EDDS-treated soil upon co-inoculation with the PGPR and rhizobium strains, which was 1.2 times higher than that without co-inoculation. Partial least squares path modeling identified plant oxidative damage and soil microbial biomass as the key variables influencing Cu uptake by alfalfa roots. Co-inoculation significantly reduced the oxidative damage to alfalfa by mitigating the accumulation of malondialdehyde and reactive oxygen species, and improving the antioxidation capacity of the plant in the presence of EDDS. EDDS application decreased microbial diversity in the rhizosphere, whereas co-inoculation increased microbial biomass carbon and nitrogen, and microbial community diversity. Increased relative abundances of Actinobacteria and Bacillus and the presence of Firmicutes taxa as potential biomarkers demonstrated that co-inoculation increased soil nutrient content, and improved plant growth. Co-inoculation with PGPR and rhizobium can be useful for altering plant-soil biochemical responses during EDDS-enhanced phytoremediation to alleviate phytotoxicity of heavy metals and improve soil biochemical activities. This study provides an effective strategy for improving phytoremediation efficiency and soil quality during chelant assisted phytoremediation of metal-contaminated soils.

Application of signaling molecules in reducing metal accumulation in alfalfa and alleviating metal-induced phytotoxicity in Pb/Cd-contaminated soil.

The utilization of forages grown on metal-contaminated soil can increase the risk of heavy metals entering the food chain and affecting human health because of elevated toxic metal concentrations. Meanwhile, hydrogen sulfide (H2S) and nitric oxide (NO) as signaling molecules are known to promote plant growth in metal-contaminated soils. However, the regulatory mechanisms of such molecules in plant physiology and soil biochemistry have not been well-documented. Hence, we investigate the role of the exogenous application of H2S and NO on alfalfa growth in lead/cadmium (Pb/Cd)-contaminated soil. Our results indicate that the signaling molecules increase the alfalfa chlorophyll and biomass content and improve alfalfa growth. Further, H2S and NO reduce the translocation and bioconcentration factors of Pb and Cd, potentially reducing the risk of heavy metals entering the food chain. These signaling molecules reduce metal-induced oxidative damage to alfalfa by mitigating reactive oxygen species accumulation and increasing antioxidant enzyme activities. Their exogenous application increases soil enzymatic activities, particularly of catalase and polyphenol oxidase, without significantly changing the composition and structure of rhizosphere bacterial communities. Interestingly, H2S addition enriches the abundance of plant-growth-promoting rhizobacteria in soil, including Nocardioides, Rhizobium, and Glycomyces. H2S is more effective than NO in improving alfalfa growth and reducing heavy-metal contamination of the food chain. These results provide new insights into the exogenous application of signaling molecules in alleviating metal-induced phytotoxicity, including an efficient strategy for the safe use of forages.

Patterns of soil microbial nutrient limitations and their roles in the variation of soil organic carbon across a precipitation gradient in an arid and semi-arid region.

The effects of precipitation patterns on the metabolism of soil microbes are poorly understood, especially in water-limited ecosystems where soil microorganisms play crucial roles in the turnover of soil organic carbon (SOC) and nutrients. We investigated the influence of the gradient levels of mean annual precipitation (MAP from 300 to 900 mm) on soil microbial metabolism in an arid and semi-arid grassland region located in Loess Plateau, China and identified relationships between microbial metabolic limitations and the variation of soil organic matter (SOM). Microbial metabolism in this arid and semi-arid region was limited by soil C and phosphorus (P) or nitrogen (N). Microbial C and P limitations decreased with the increase of MAP. Microbial C and P limitations were lowest in the areas with MAPs of 700-900 mm, whereas N limitation was observed in the areas with MAPs >700 mm. The results of a variation-partitioning analysis and partial least squares path modeling indicated that the microbial C and N/P limitations on regional scales were mainly determined by climate factors (MAP and mean annual temperature (MAT)), followed by vegetation biomass and soil properties. The extents of soil drying-rewetting processing caused by different MAPs directly affected microbial nutrient limitation. Our results suggested that the influence of precipitation variation on microbial metabolic limitation strongly governed SOM stability and that an increase in the rate of SOM decomposition with increasing precipitation could be caused by increased microbial nutrient limitation. SOM may be most stable at a MAP of 700 mm in the arid and semi-arid regions (300-900 mm MAP) where microbial nutrient limitation was lowest. This study provided novel insights into the responses of soil microbial metabolism to precipitation change and is an important step toward understanding the mechanisms of SOM stability in an arid and semi-arid grassland ecosystem under scenarios of precipitation variation.

Natural grassland as the optimal pattern of vegetation restoration in arid and semi-arid regions: Evidence from nutrient limitation of soil microbes.

Soil microbial metabolism is vital for nutrient cycling and aboveground ecosystem stability. A general understanding of microbial metabolism and nutrient limitation under human disturbance in arid and semi-arid regions, which are the largest and most fragile oligotrophic ecosystems globally, however, is still limited. We quantified and compared the characteristics of nutrient limitation of soil microbes under natural/artificial grassland and shrubland, an ecological forest, an economic forest, and sloped cropland in typical arid and semi-arid ecosystems on the Loess Plateau, China. Vegetation restoration significantly affected the activities of extracellular enzymes and ecoenzymatic stoichiometry mainly by affecting soil nutrients and nutrient stoichiometry. A vector analysis of enzyme activity indicated that microbial communities were co-limited by carbon (C) and phosphorus (P) in all types of vegetation restoration. Linear regression indicated that microbial C and P limitations were significantly correlated with the stoichiometry of soil nutrient, suggesting that the balance of nutrient stoichiometry is an important factor maintaining microbial metabolism and elemental homeostasis. C and P limitations in the microbial communities were the lowest in the natural grassland. This implies that both vegetation and microbial communities under the restoration pattern of natural grassland were more stable under environmental stress, so the restoration of natural grassland should be recommended as the preferred option for ecosystem restoration in these arid and semi-arid regions.