Search

Aims Revegetation is an effective measure to improve soil structure and nutrients in erosion-prone areas. However, little is known about the impact of diverse phytomanagement of revegetation on soil quality and ecological security near tailings.Methods We investigated the water-stability and soil aggregate nutrients and assessed the associated risk of heavy metal contamination under multiple phytomanagement (natural grassland, artificial forest, and artificial forest mixed with shrubs and herbs) near abandoned tailings on the Loess Plateau, comparing with the adjacent bare land.Results The results showed that phytomanagement significantly enhanced soil aggregate stability, as demonstrated by the rise in soil macro-aggregate proportion (> 0.25 mm, 10.5–22.4%) and geometric mean diameter (16.3–44.0%). Furthermore, the soil organic carbon content (SOC), glomalin-related soil protein (GRSP), aromatic-C, and alkene-C in macro-aggregates increased alongside aggregate stability enhancement. The increased stability of soil aggregates following phytomanagement could reduce the risk of heavy metal leaching, but the increased concentration of heavy metals in the aggregates. In addition, the management of artificial forests mixed with shrubs and herbs greatly reduced the ecological risk of heavy metal pollution compared to other phytomanagement. SOC and GRSP were identified as key factors influencing the risk of heavy metal contamination in soil aggregates following phytomanagement.Conclusion Our study highlights that revegetation of tailings improves soil quality and ecological security by increasing the stability of soil structure as well as SOC and GRSP within the aggregates. Artificial forests mixed with shrubs and herbs could be an optimal phytomanagement to achieve environmental sustainability in tailings areas.

Tea gardens, being a key agroecosystem type, are an important source of nitrous oxide (N2O) emissions. However, main factors that regulate N2O emissions following urea and organic matter amendments have yet to be clarified. To investigate the influence of different fertilization management measures on N2O emissions in tea garden soil, a 50-day laboratory incubation experiment was conducted. Five treatments were designed for this experiment: control (CK), urea (U), rapeseed cake (R), urea + rapeseed cake (2:1, UR1), and urea + rapeseed cake (1:2, UR2). N2O emission flux in the R treatment peaked at 14.12 μg kg−1 h−1 on day 19, which occurred later than the UR1, UR2, and U treatments. Cumulative N2O emissions from the R and UR2 treatments were 6073 and 4296 μg kg−1, respectively, which were greater by a factor of 11.2 and 7.9, respectively, compared to the U treatment. Moreover, N2O emissions of the UR1 and UR2 treatments were significantly lower than the R treatment. Additionally, N2O emissions were also significantly positively correlated to pH levels and microbial biomass carbon (MBC) content. MBC content had the most direct and greatest influence on soil N2O emissions, indicating that MBC could be the key limiting factor for N2O emissions in this experiment. A single application of rapeseed cake caused an increase in N2O emissions, whereas the combined application of rapeseed cake and a synthetic N fertilizer (urea) caused a decrease in N2O emissions. Results from this study offer potential strategies to mitigate soil N2O emissions from tea garden agroecosystems through improved field fertilization management.

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.

Agricultural practices such as fertilization considerably influence soil greenhouse gas fluxes. However, the effects of fertilization on greenhouse gases fluxes remain unclear in tea soil when soil nitrogen is low. In the present study, soil CO2 and CH4 fluxes under various fertilization treatments in tea soil were investigated during a 50-day period. The experiment consisted of five treatments: no fertilizer (CK), single nitrogen (urea, N), single oilseed rape cake fertilizer (R), nitrogen + cake fertilizer (2:1, NR1), and nitrogen + cake fertilizer (1:2, NR2). The fertilization proportion of NR1 and NR2 was determined by the nitrogen content of nitrogen fertilizer and cake fertilizer. The results revealed that the single application of nitrogen had no significant effect on soil CO2 flux. However, the addition of cake fertilizer significantly increased CO2 emissions through enhanced soil microbial biomass carbon (MBC). Additionally, CO2 emissions were directly proportional to the amount of carbon (C) in the fertilizer. All treatments were minor sinks for CH4 except for the treatment NR1. Specifically, the cumulative CH4 fluxes of NR1 and NR2 were significantly higher than rest of the three treatments, which implies that application of urea and oilseed rape cake reduced the capability of CH4 oxidation in tea soil. Structural equation models indicated that soil CO2 flux is significantly and positively correlated with soil dissolved organic carbon, MBC and soil pH, while mineral nitrogen content was the main factor affecting CH4 flux. Overall, the application of oilseed rape cake increased the oxidation of CH4 and promoted soil C sequestration but inevitably increased the soil CO2 emissions.

Plant growth-promoting rhizobacteria (PGPR) and rhizobia are potentially advantageous in improving plant growth in heavy metal contaminated soils. However, only limited information is available in literature on the manner through which the co-inoculation of PGPR and rhizobia can potentially supply nutrients to benefit plant growth in heavy metal contaminated soil. Accordingly, this study investigated the effects of Paenibacillus mucilaginosus (PGPR) and Sinorhizobium meliloti (rhizobia) co-inoculation on soil nutrients, enzyme activities, and microbial biomass in copper (Cu) contaminated soil planted with alfalfa (Medicago sativa). Moreover, we assessed soil bacterial community structure using high-throughput Illumina sequencing of 16S rRNA genes. Results showed that PGPR and/or rhizobia inoculation improved alfalfa growth. In particular, we found that this co-inoculation approach decreased Cu accumulation (48.6%) in shoots compared to the control (uninoculated). Both partial least squares path modeling (PLS-PM) and the relative importance of regressors in the linear models identified that enzyme activities, microbial biomass, and microbial community structure in Cu contaminated soil were major controlling variables of soil nutrient availability. The co-inoculation treatment significantly increased soil carbon (C) and nitrogen (N) concentrations by increasing urease (55.6%), saccharase (29.5%), and β-glucosidase (31.4%) activities compared to the control. Furthermore, the rhizosphere microbial community structure in the co-inoculation treatment was mainly regulated by soil N concentrations (i.e., both total N and available N) while altering alpha diversity (α-diversity). The relative abundances of Firmicutes (including biomarkers of the Bacillus genus) and Acidobacteria were enriched in the co-inoculated treatment, which can potentially improve soil nutrient availability and subsequently benefit plant growth. These findings indicated that the co-inoculation of PGPR and rhizobia plays an important role in promoting plant growth in Cu contaminated soil. This is because this approach can increase soil nutrient availability by enhancing soil enzyme activities and regulating rhizosphere microbial community structure.

The effects and regulatory mechanisms of co-inoculation of plant-growth-promoting rhizobacteria (PGPRs) and rhizobium in plant-soil systems remain unclear, despite numerous reports that PGPRs or rhizobium can alleviate metal toxicity. We used the co-inoculation of the PGPR Paenibacillus mucilaginosus and the metal-resistant rhizobium Sinorhizobium meliloti for exploring the physiological and biochemical responses of the plant-soil system in metal-contaminated soil. The co-inoculation with the PGPR and rhizobium significantly increased the nutrient (N, P, and K) contents in plant tissues and promoted plant growth in soil contaminated with copper (Cu). Stress from Cu-induced reactive oxygen species and lipid peroxidation were largely attenuated by the co-inoculation by increasing the activities of antioxidant enzymes. The contents and uptake of Cu in plant tissues increased significantly in the co-inoculation treatment compared with the uninoculated control and individual inoculation treatment. Co-inoculation with PGPR and rhizobium significantly increased soil microbial biomass, enzymatic activities, total nitrogen, available phosphorus, and soil organic matter contents compared with the uninoculated control. Interestingly, co-inoculation also affected the composition of the rhizospheric microbial community, and slightly increased rhizospheric microbial diversity. These improvements of the soil fertility and biological activity also had a beneficial impact on plant growth under Cu stress. Our results suggested that alfalfa co-inoculated with PGPR and rhizobium could increase plant growth and Cu uptake in metal-contaminated soil by alleviating plant Cu stress and improving soil biochemical properties. These results indicate that the co-application of PGPR and rhizobium can have a positive effect on the biochemical responses of alfalfa-soil systems in soil contaminated by heavy metals and can provide an efficient strategy for the phytomanagement of metal-contaminated land.

Being signaling molecules, nitric oxide (NO) and hydrogen sulfide (H2S) can mediate a wide range of physiological processes caused by plant metal toxicity. Moreover, legume-rhizobium symbiosis has gained increasing attention in mitigating heavy metal stress. However, systematic regulatory mechanisms used for the exogenous application of signaling molecules to alter the resistance of legume-rhizobium symbiosis under metal stress are currently unknown. In this study, we examined the exogenous effects of sodium nitroprusside (SNP) as an NO donor additive and sodium hydrosulfide (NaHS) as a H2S donor additive on the phytotoxicity and soil quality of alfalfa (Medicago sativa)-rhizobium symbiosis in lead/cadmium (Pb/Cd)-contaminated soils. Results showed that rhizobia inoculation markedly promoted alfalfa growth by increasing chlorophyll content, fresh weight, and plant height and biomass. Compared to the inoculated rhizobia treatment alone, the addition of NO and H2S significantly reduced the bioaccumulation of Pb and Cd in alfalfa-rhizobium symbiosis, respectively, thus avoiding the phytotoxicity caused by the excessive presence of metals. The addition of signaling molecules also alleviated metal-induced phytotoxicity by increasing antioxidant enzyme activity and inhibiting the level of lipid peroxidation and reactive oxygen species (ROS) in legume-rhizobium symbiosis. Also, signaling molecules improved soil nutrient cycling, increased soil enzyme activities, and promoted rhizosphere bacterial community diversity. Both partial least squares path modeling (PLS-PM) and variation partitioning analysis (VPA) identified that using signaling molecules can improve plant growth by regulating major controlling variables (i.e., soil enzymes, soil nutrients, and microbial diversity/plant oxidative damage) in legume-rhizobium symbiosis. This study offers integrated insight that confirms that the exogenous application of signaling molecules can enhance the resistance of legume-rhizobium symbiosis under metal toxicity by regulating the biochemical response of the plant-soil system, thereby minimizing potential health risks.

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.

Grazing prohibition is an effective measure in improving soil stability and ecological quality. However, only a limited number of studies have been published on the dominant factors that impact soil aggregate stability and their associated effects on nutrient distribution for different size soil aggregates under long-term grazing prohibition management. In this study, we investigated variation in soil aggregate stability and nutrient distribution characteristics in semiarid grassland sites under different grazing prohibition timeframes (0 years [GP0], 11 years [GP11], 26 years [GP26], and 36 years [GP36]). Results showed that organic carbon (C) and total nitrogen (TN) concentrations in soil aggregates decreased at GP11 before progressively increasing and reaching its highest value at GP36, and the total phosphorus (TP) concentration did not change significantly. Most nutrients accumulated in macroaggregates (> 0.25 mm) under grazing prohibition, and the nutrient stoichiometry in soil aggregates increased after 26 years. Compared to the control (GP0), the mean weight diameter (MWD) value of the soil stability index increased at GP11 (21.7%) and decreased at GP26 (18.9%). Attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) also showed that the proportion of stable organic C-related functional groups (i.e., alkene-C and aromatic-C) in macroaggregates were higher at GP11 and GP36 than at GP26. Furthermore, principal component analysis (PCA), partial least squares path modeling (PLS-PM), and the relative importance of regressors all showed that glomalin-related soil proteins (GRSP) and nutrients indirectly improved aggregate stability in semiarid grassland through their influence on the GRSP accumulation potential and nutrient stoichiometry. Generally, after 26 years grazing prohibition had a positive effect on soil aggregate stability and nutrient accumulation in the semiarid grassland sites investigated for this study. Results from this study provide a theoretical basis to select appropriate grazing prohibition timeframes under grassland management initiatives to optimize ecological quality measures in semiarid regions.

The application of green manure is a traditional and valuable practice for agroecosystem management. In northern China, the effects of green manure on production of the region’s major crops have been extensively investigated, but the inconsistent conclusions that these case studies have yielded cannot provide effective guidance for practical local agricultural production. Here, we conducted a meta-analysis to generate a comprehensive evaluation of the effects of green manure on soil properties and crop yield in this region. Our results shown that green manure improves soil quality effectively, decreasing soil bulk density by ∼ 5.6 %, increasing microbial biomass carbon by 28 %, and improving the activities of soil enzymes by 14 % ∼ 39 %. Among the different types of green manure, legume green manure more markedly increased both nitrate and hydrolysable nitrogen, while non-legume green manure more markedly increased available potassium. Soil gravimetric water content was decreased under green manure treatment. Maize yield was significantly increased under green manure by 11 % on the whole, while effects of green manure on wheat and potato were inconsistent. In summary, the application of green manure in northern China can improve soil quality significantly, and proper green manure use can improve cash crop yields.

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.

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.

Revegetation of semiarid lands depends upon soil microbial communities to supply nutrients for successive plant species, but microbial activity can be constrained by insufficient water. The objective of this study was to quantify the metabolic limitation of microbes by extracellular enzymatic stoichiometry, and to determine how this affected microbial carbon use efficiency (CUE) with biogeochemical equilibrium model. The study occurred in long-term revegetation experiment with seven successional stages (0, 11, 35, 60, 100, 130 and 150 years) in the Loess Plateau, China. Microbes maintained stoichiometric homeostasis in all successional stages, but plants did not. Microbial metabolism was limited by low soil phosphorus (P) concentration throughout the succession, whereas plants were limited by low soil P during the late successional stages (from 60 to 150 years) only. An increase in soil moisture during succession was associated with greater P limitation in microbes and plants. There was less microbial P limitation at the 35-year successional stage, and the greatest microbial P limitation occurred at the 130-year successional stage. The microbial C limitation followed a unimodal pattern through the vegetation succession and reached a maximum at 100 years of succession (the early forest stage). This coincided with the lowest microbial CUE at 100 years of succession (CUE was from 0.24 to 0.41), suggesting a change in the physiological responses from microbes (such as enzyme synthesis and the priming effect), that tended to reduce soil C sequestration. Our results indicate that soil moisture regulated microbial C and P metabolism during the vegetation succession in this semiarid region, which has implications for understanding how microbial metabolism affects soil C dynamics under vegetation restoration.

Legume-rhizobium symbiosis has been heavily investigated for their potential to enhance plant metal resistance in contaminated soil. However, the extent to which plant resistance is associated with the nitrogen (N) supply in symbiont is still uncertain. This study investigates the effect of urea or/and rhizobium (Sinorhizobium meliloti) application on the growth of Medicago sativa and resistance in metals contaminated soil (mainly with Cu). The results show that Cu uptake in plant shoots increased by 41.7%, 69%, and 89.3% with urea treatment, rhizobium inoculation, and their combined treatment, respectively, compared to the control group level. In plant roots, the corresponding values were 1.9-, 1.7-, and 1.5-fold higher than the control group values, respectively. Statistical analysis identified that N content was the dominant variable contributing to Cu uptake in plants. Additionally, a negative correlation was observed between plant oxidative stress and N content, indicating that N plays a key role in plant resistance. Oxidative damage decreased after rhizobium inoculation as the activities of antioxidant enzymes (catalase and superoxide dismutase in roots and peroxidase in plant shoots) were stimulated, enhancing plant resistance and promoting plant growth. Our results suggest that individual rhizobium inoculation, without urea treatment, is the most recommended approach for effective phytoremediation of contaminated land.

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.

Variations in soil microbial metabolism currently represent one of the greatest areas of uncertainty with regard to soil nutrient cycles and the control of terrestrial carbon (C) and nitrogen (N) loss and are poorly understood in agricultural ecosystems with intensive farming practices. In this study, extracellular enzymatic stoichiometry models and quantitative PCR techniques were used to examine microbial metabolic limitation and its relationship with N-cycling gene expression in semi-arid agricultural ecosystems considering four N fertilization levels (N 0, N 100, N 250, and N 400 kg N ha−1) and two agronomic strategies (film mulching and no mulching). Film mulching increased microbial C limitation (reflecting microbial C metabolism size; 0.189 of the total effects), while very small effects on microbial phosphorus (P) limitation were observed (-0.007 of the total effects). N fertilization increased the microbial demand for P (microbial P limitation; 0.504 of the total effects). Increased microbial C metabolism was mainly attributed to increased soil moisture content after film mulching, which enhanced microbial decomposition of organic C (high C-acquiring enzyme activities). Changes in nutrient stoichiometry and the increase in N availability due to N fertilization were largely responsible for increased microbial P limitation. Furthermore, microbial P limitation negatively affected the abundance of AOA amoA, AOB amoA (involved in nitrification), nirK, nirS, nosZ (involved in denitrification) genes, strongly inhibiting nitrification and denitrification potential (-0.743 and -0.761 of the total effects, respectively). The present results suggest that agricultural ecosystems with film mulching are conducive to organic residue decomposition, while appropriate P limitation under N fertilization could reduce the loss of N due to nitrification and denitrification in soil. This study highlights the importance of elemental stoichiometry-driven microbial metabolic variation in understanding soil nutrient cycles and optimizing agricultural practices.

Legume–rhizobium symbiosis plays an important role in agriculture and ecological restoration. However, the regulatory mechanisms of rhizobium in alleviating heavy metal stress through the biochemical response of plant-soil system is limited. In this study, alfalfa was inoculated with a copper (Cu)-resistant rhizobium, and its effect on plant growth and the spatial distribution of phosphatase in the rhizosphere under Cu stress was assessed. Our results showed that rhizobium inoculation alleviated Cu-induced growth inhibition, and increased the nitrogen and phosphorus content in alfalfa tissues. Moreover, inoculated plants had a higher Cu uptake than non-inoculated plants, with a much higher increase in the roots than in the shoots; thus, inoculation with rhizobium was shown to decrease the transfer coefficient and promote Cu phytostabilization. The zymograms illustrated that the distribution of phosphatase activities was associated with the presence of roots. Compared with the non-inoculated treatment, the rhizobium inoculation increased the hotspot areas of phosphatase by 26.1% and 39.3% at the Cu 0 and Cu 800 treatments, respectively. In addition, the available phosphorus in the soil showed negative correlations with soil phosphatase activity (p