Effects of soil amendment with different carbon sources and other factors on the bioremediation of an aged PAH-contaminated soil

Carbon supplementation, soil moisture and soil aeration are believed to enhance in situ bioremediation of PAH-contaminated soils by stimulating the growth of indigenous microorganisms. However, the effects of added carbon and nitrogen together with soil moisture and soil aeration on the dissipation of PAHs and on associated microbial counts have yet to be fully assessed. In this study the effects on bioremediation of carbon source, carbon-to-nitrogen ratio, soil moisture and aeration on an aged PAH-contaminated agricultural soil were studied in microcosms over a 90-day period. Additions of starch, glucose and sodium succinate increased soil bacterial and fungal counts and accelerated the dissipation of phenanthrene and benzo(a)pyrene in soil. Decreases in phenanthrene and benzo(a)pyrene concentrations were effective in soil supplemented with glucose and sodium succinate (both 0.2 g C kg⁻¹ dry soil) and starch (1.0 g C kg⁻¹ dry soil). The bioremediation effect at a C/N ratio of 10:1 was significantly higher (P < 0.05) than at a C/N of either 25:1 or 40:1. Soil microbial counts and PAH dissipation were lower in the submerged soil but soil aeration increased bacterial and fungal counts, enhanced indigenous microbial metabolic activities, and accelerated the natural degradation of phenanthrene and benzo(a)pyrene. The results suggest that optimizing carbon source, C/N ratio, soil moisture and aeration conditions may be a feasible remediation strategy in certain PAH contaminated soils with large active microbial populations.     

Efficacy of bacterial consortium-AIE2 for contemporaneous Cr(VI) and azo dye bioremediation in batch and continuous bioreactor systems, monitoring steady-state bacterial dynamics using qPCR assays

Bacterial consortium-AIE2 with a capability of contemporaneous Cr(VI) reduction and azo dye RV5 decolourization was developed from industrial wastewaters by enrichment culture technique. The 16S rRNA gene based molecular analyses revealed that the consortium bacterial community structure consisted of four bacterial strains namely, Alcaligenes sp. DMA, Bacillus sp. DMB, Stenotrophomonas sp. DMS and Enterococcus sp. DME. Cumulative mechanism of Cr(VI) reduction by the consortium was determined using in vitro Cr(VI) reduction assays. Similarly, the complete degradation of Reactive Violet 5 (RV5) dye was confirmed by FTIR spectroscopic analysis. Consortium-AIE2 exhibited simultaneous bioremediation efficiencies of (97.8 ± 1.4) % and (74.1 ± 1.2) % in treatment of both 50 mg l⁻¹ Cr(VI) and RV5 dye concentrations within 48 h of incubation at pH 7 and 37°C in batch systems. Continuous bioreactor systems achieved simultaneous bioremediation efficiencies of (98.4 ± 1.5) % and (97.5 ± 1.4) % after the onset of steady-state at 50 mg l⁻¹ input Cr(VI) and 25 mg l⁻¹ input RV5 concentrations, respectively, at medium dilution rate (D) of 0.014 h⁻¹. The 16S rRNA gene copy numbers in the continuous bioreactor as determined by real-time PCR assay indicated that Alcaligenes sp. DMA and Bacillus sp. DMB dominated consortium bacterial community during the active continuous bioremediation process.     

Application of the knowledge-based approach to strain selection for a bioaugmentation process of phenanthrene- and Cr(VI)-contaminated soil

Aims: The objective of this study was to apply the knowledge‐based approach to the selection of an inoculum to be used in bioaugmentation processes to facilitate phenanthrene degradation in phenanthrene‐ and Cr(VI)‐co‐contaminated soils. Methods and Results: The bacterial community composition of phenanthrene and phenanthrene‐ and Cr(VI)‐co‐contaminated microcosms, determined by denaturing gradient gel electrophoresis analysis, showed that members of the Sphingomonadaceae family were the predominant micro‐organisms. However, the Cr(VI) contamination produced a selective change of predominant Sphingomonas species, and in co‐contaminated soil microcosms, a population closely related to Sphingomonas paucimobilis was naturally selected. The bioaugmentation process was carried out using the phenanthrene‐degrading strain S. paucimobilis 20006FA, isolated and characterized in our laboratory. Although the strain showed a low Cr(VI) resistance (0·250 mmol l^?1); in liquid culture, it was capable of reducing chromate and degrading phenanthrene simultaneously. Conclusion: The inoculation of this strain managed to moderate the effect of the presence of Cr(VI), increasing the biological activity and phenanthrene degradation rate in co‐contaminated microcosm. Significance and Impact of the Study: In this study, we have applied a novel approach to the selection of the adequate inoculum to enhance the phenanthrene degradation in phenanthrene‐ and Cr(VI)‐co‐contaminated soils.     

Soil chromium bioremediation: Synergic activity of actinobacteria and plants

The aim of this work was to evaluate a strategy to reduce the bioavailable chromium fraction in soil, using a Cr(VI) resistant microorganism, Streptomyces sp. MC1, under non sterile conditions, with maize plants as bioindicator and/or bioremediator.Soil samples were contaminated with 100, 200 and 400 mg kg⁻¹ of Cr(VI) or Cr(III). Bioavailable chromium (35%) was only detected in samples with Cr(VI). Soil samples with Cr(VI) 200 mg kg⁻¹ were inoculated with Streptomyces sp. MC1, and bioavailable chromium decreased up to 73%.Zea mays seedlings were planted in soil samples contaminated with chromium. Plantlets accumulated chromium mainly as Cr(III), and biomass decreased up to 88%. Streptomyces sp. MC1 was inoculated in soil samples contaminated with 200 mg kg⁻¹ of Cr(VI) and Z.mays seedlings were planted.Streptomyces sp. MC1 caused Z.mays biomass increase (57%), chromium accumulation and bioavailable chromium decreased up to 46% and 96%, respectively.This work constitutes the first contribution of cooperative action between actinobacteria and Z.mays in the bioremediation of Cr(VI) contaminated soil. The large removal capacity of bioavailable chromium by Streptomyces sp. MC1 and Z.mays infers that they could be successfully applied together in bioremediation of soils contaminated with Cr(VI).     

Evaluation of the bioremoval of Cr(VI) and TOC in biofilters under continuous operation using response surface methodology

In the present study, the bioremoval of Cr(VI) and the removal of total organic carbon (TOC) were achieved with a system composed by an anaerobic filter and a submerged biofilter with intermittent aeration using a mixed culture of microorganisms originating from contaminated sludge. In the aforementioned biofilters, the concentrations of chromium, carbon, and nitrogen were optimized according to response surface methodology. The initial concentration of Cr(VI) was 137.35 mg l⁻¹, and a bioremoval of 85.23% was attained. The optimal conditions for the removal of TOC were 4 to 8 g l⁻¹ of sodium acetate, >0.8 g l⁻¹ of ammonium chloride and 60 to 100 mg l⁻¹ of Cr(VI). The results revealed that ammonium chloride had the strongest effect on the TOC removal, and 120 mg l⁻¹ of Cr(VI) could be removed after 156 h of operation. Moreover, 100% of the Cr(VI) and the total chromium content of the aerobic reactor output were removed, and TOC removals of 80 and 87% were attained after operating the anaerobic and aerobic reactors for 130 and 142 h, respectively. The concentrations of cells in both reactors remained nearly constant over time. The residence time distribution was obtained to evaluate the flow through the bioreactors.     

A bioprocessing strategy that allows for the selection of Cr(VI)-reducing bacteria from soils

Anaerobic bacteria that reduce hexavalent chromium [Cr(VI)] to trivalent [Cr(III)] are common in soils and were used to develop a bioprocess employing a selection strategy. Indigenous Cr(VI)-reducers were enriched from Cr(VI)-contaminated soil under anaerobic conditions. The mixed culture was then tested for Cr(VI)-reducing activity in a chemostat, followed by transfer to a 1-L packed-bed bioreactor operated at 30°C for additional study. The support material used in the reactor consisted of 6-mm porcelain saddles. Cr(VI) concentrations in the liquid ranged from 140–750 mg L⁻¹. Cr(VI)-reducing bacteria were the dominant population with Cr(VI)-reduction rates of approximately 0.71 mg g⁻¹ dry cells h⁻¹ achieved at Cr(VI) concentrations of 750 mg L⁻¹. These results indicate a potential for selecting and maintaining indigenous Cr(VI)-reducers in a bioreactor for Cr(VI)-remediation of groundwater or soil wash effluents. Received 09 January 1996/ Accepted in revised form 15 November 1996     

Enhancement of hexavalent chromium reduction and electricity production from a biocathode microbial fuel cell

Enhancement of Cr (VI) reduction rate and power production from biocathode microbial fuel cells (MFCs) was achieved using indigenous bacteria from Cr (VI)-contaminated site as inoculum and MFC architecture with a relatively large cathode-specific surface area of 340–900 m² m⁻³. A specific Cr (VI) reduction rate of 2.4 ± 0.2 mg g⁻¹VSS h⁻¹ and a power production of 2.4 ± 0.1 W m⁻³ at a current density of 6.9 A m⁻³ were simultaneously achieved at an initial Cr (VI) concentration of 39.2 mg L⁻¹. Initial Cr (VI) concentration and solution conductivity affected Cr (VI) reduction rate, power production and coulombic efficiency. These findings demonstrate the importance of inoculation and MFC architecture in the enhancement of Cr (VI) reduction rate and power production. This study is a beneficial attempt to improve the efficiency of biocathode MFCs and provide a good candidate of bioremediation process for Cr (VI)-contaminated sites.     

Identification and hexavalent chromium reduction characteristics of <Emphasis Type="Italic">Pannonibacter phragmitetus</Emphasis>

A hexavalent chromium [Cr(VI)] reducing bacterial strain was isolated from chromium-containing slag. It was identified as Pannonibacter phragmitetus based on physiological, biochemical characteristics and 16S rRNA gene sequence analysis. This bacterium displayed great Cr(VI) reduction capability. The Cr(VI) could be completely removed in 24 h under anaerobic condition when the initial concentration was 1,917 mg L⁻¹, with the maximum reduction rate of 562.8 mg L⁻¹ h⁻¹. The Cr(VI) reduction rate increased with the increase of Cr(VI) concentration. P. phragmitetus was able to use many carbon sources such as lactose, fructose, glucose, pyruvate, citrate, formate, lactate, NADPH and NADH as electron donors, among which the lactate had the greatest power to promote the reduction process. Zn²⁺, Cd²⁺ and Ni²⁺ inhibited, while Cu²⁺, Pb²⁺, Mn²⁺ and Co²⁺ stimulated the reduction. The optimum pH and temperature for reduction were 9.0 and 30 °C, respectively. The results indicated that this strain had great potential for application in the bioremediation of chromate-polluted soil and water systems.     

Isolation and genetic identification of PAH degrading bacteria from a microbial consortium

Polycyclic aromatic hydrocarbons (PAH; naphthalene, anthracene and phenanthrene) degrading microbial consortium C2PL05 was obtained from a sandy soil chronically exposed to petroleum products, collected from a petrochemical complex in Puertollano (Ciudad Real, Spain). The consortium C2PL05 was highly efficient degrading completely naphthalene, phenanthrene and anthracene in around 18 days of cultivation. The toxicity (Microtox™ method) generated by the PAH and by the intermediate metabolites was reduced to levels close to non-toxic in almost 40 days of cultivation. The identified bacteria from the contaminated soil belonged to γ-proteobacteria and could be include in Enterobacter and Pseudomonas genus. DGGE analysis revealed uncultured Stenotrophomonas ribotypes as a possible PAH degrader in the microbial consortium. The present work shows the potential use of these microorganisms and the total consortium for the bioremediation of PAH polluted areas since the biodegradation of these chemicals takes place along with a significant decrease in toxicity.     

Permissible Value for Vanadium in Allitic Udic Ferrisols Based on Physiological Responses of Green Chinese Cabbage and Soil Microbes

Greenhouse experiments were conducted to study the permissible value of vanadium (V) based on the growth and physiological responses of green Chinese cabbage (Brassica chinensis L.), and effects of V on microbial biomass carbon (MBC) and enzyme activities in allitic udic ferrisols were also studied. The results showed that biomass of cabbage grown on soil treated with 133 mg V kg⁻¹ significantly decreased by 25.1% compared with the control (P < 0.05). Vanadium concentrations in leaves and roots increased with increasing soil V concentration. Contents of vitamin C (Vc) increased by 10.3%, while that of soluble sugar in leaves significantly decreased by 54.0% when soil V concentration was 133 mg kg⁻¹, respectively. The uptake of essential nutrient elements by cabbage was disturbed when soil V concentration exceeded 253 mg kg⁻¹. Soil MBC was significantly stimulated by 15.5%, while dehydrogenase activity significantly decreased by 62.8% and urease activity slightly changed at treatment of 133 mg V kg⁻¹ as compared with the control, respectively. Therefore, the permissible value of V in allitic udic ferrisols is proposed as 130 mg kg⁻¹.     

Effect of decabromodiphenyl ether (BDE 209) on soil microbial activity and bacterial community composition

In many areas of the world, polybrominated diphenyl ethers are ubiquitous due to their use as fire retardants. It is known that the hydrophobic characteristics of PBDEs cause them to sink in soil and sediment, yet their effect on microbes within the soil is not well understood. In this study, soil was treated with 1, 10, and 100 mg kg⁻¹ BDE 209 for up to 45 days. Treatment effects on soil enzymatic activities for urease and catalase were evaluated. The impact on the microbe community structure was estimated using denaturing gradient gel electrophoresis after polymerase chain reaction amplification of total genomic DNA, using bacterial variable V3 region targeted primers. The effects on the soil microbial community size and major bacterial groups were evaluated using fluorescence in situ hybridization analysis. Forty-five days after the addition of BDE 209, urease activity was suppressed by BDE 209, even at a concentration of 1 mg kg⁻¹. Catalase activity increased in the samples containing lower concentrations of BDE 209, but was suppressed in samples containing higher concentrations. The bacterial community also varied in response to the addition of BDE 209, and the variation of community composition differed among treatments. In addition, α, β and γ subclass proteobacteria decreased in the group of 100 mg kg⁻¹ BDE 209 spiked soil after 45 days of treatment. Throughout the experiment, no BDE 209 degradation was detected under darkness. These observations demonstrated that BDE 209 in soil, although of low bioavailability, had an adverse impact on the structure and function of the soil microbial community and microbial processes.     

Characterization of polycyclic aromatic hydrocarbons degradation and arsenate reduction by a versatile Pseudomonas isolate

A Pseudomonas isolate, designated PAHAs-1, was found capable of reducing arsenate and degrading polycyclic aromatic hydrocarbons (PAHs) independently and simultaneously. This isolate completely reduced 1.5 mM arsenate within 48 h and removed approximately 100% and 50% of 60 mg l⁻¹ phenanthrene and 20 mg l⁻¹ pyrene within 60 h, respectively. Using PAHs as the sole carbon source, however, this isolate showed a slow arsenate reduction rate (4.62 μM h⁻¹). The presence of arsenic affected cell growth and concurrent PAHs removal, depending on PAH species and arsenic concentration. Adding sodium lactate to the medium greatly enhanced the arsenate reduction and pyrene metabolism. The presence of the alpha subunit of the aromatic ring-hydroxylating dioxygenase (ARHD) gene, arsenate reductase (arsC) and arsenite transporter (ACR3(2)) genes supported the dual function of the isolate. The finding of latter two genes indicated that PAHAs-1 possibly reduced arsenate via the known detoxification mechanism. Preliminary data from hydroponic experiment showed that PAHAs-1 degraded the majority of phenanthrene (>60%) and enhanced arsenic uptake by Pteris vittata L. (from 246.7 to 1187.4 mg kg⁻¹ As in the fronds). The versatile isolate PAHAs-1 may have potentials in improving the bioremediation of PAHs and arsenic co-contamination using the plant-microbe integrated strategy.     

Characterization of a strain of <Emphasis Type="Italic">Pseudomonas putida</Emphasis> isolated from agricultural soil that degrades cadusafos (an organophosphorus pesticide)

Bacteria capable of degrading the pesticide, cadusafos, were isolated from agricultural soil using an enrichment method. In this way, five distinct cadusafos-degrading strains of Pseudomonas putidia were isolated, and were characterized using morphological and biochemical analysis, as well as 16S rRNA sequencing. Strain PC1 exhibited the greatest cadusafos degradation rate and was consequently selected for further investigation. Degradation of cadusafos by strain PC1 was rapid at 20 and 37°C, but was greatly reduced (~1.5-fold) by the presence of carbon sources. Strain PC1 was able to effectively degrade cadusafos in sterilized soil using low inoculum levels. The maximum degradation rate of cadusafos (V _max ) was calculated as 1.1 mg l⁻¹ day⁻¹, and its saturation constant (K _s ) was determined as 2.5 mg l⁻¹. Bacteria such as strain PC1, that use cadusafos as a carbon source, could be employed for the bioremediation of sites contaminated with pesticides.     

Behavior of mercury in a soil–plant system as affected by inoculation with the arbuscular mycorrhizal fungus Glomus mosseae

Effects of inoculation with the arbuscular mycorrhizal (AM) fungus Glomus mosseae on the behavior of Hg in soil–plant system were investigated using an artificially contaminated soil at the concentrations of 0, 1.0, 2.0, and 4.0 mg Hg kg⁻¹. Mercury accumulation was lower in mycorrhizal roots than in nonmycorrhizal roots when Hg was added at the rates of 2.0 and 4.0 mg kg⁻¹, while no obvious difference in shoot Hg concentration was found between mycorrhizal and nonmycorrhizal treatments. Mycorrhizal inoculation significantly decreased the total and extractable Hg concentrations in soil as well as the ratio of extractable to total Hg in soil. Equilibration sorption of Hg by soil was investigated, and the results indicated that mycorrhizal treatment enhanced Hg sorption on soil. The uptake of Hg was lower by mycorrhizal roots than by nonmycorrhizal roots. These experiments provide further evidence for the role of mycorrhizal inoculation in increasing immobilization of Hg in soil and reducing the uptake of Hg by roots. Calculation on mass balance of Hg in soil suggests the presence of Hg loss from soil presumably through evaporation, and AM inoculation enhanced Hg evaporation. This was evidenced by a chamber study to detect the Hg evaporated from soil.     

Sulphur protects mustard (<Emphasis Type="Italic">Brassica campestris</Emphasis> L.) from cadmium toxicity by improving leaf ascorbate and glutathione

In a pot-soil culture ameliorative effect of sulphur (S) (0 or 40 mg S kg⁻¹ soil) on cadmium (Cd) (0, 25, 50 and 100 mg Cd kg⁻¹ soil)-induced growth inhibition and oxidative stress in mustard (Brassica campestris L.) cultivar Pusa Gold was studied. Cadmium at 100 mg kg⁻¹ soil caused maximum increase in the contents of Cd and thiobarbituric acid reactive substances (TBARS) in leaves. Maximum reductions in growth (plant dry mass, leaf area), chlorophyll content, net photosynthetic rate (P_N) and the contents of ascorbate (AsA), glutathione (GSH) were observed with 100 mg Cd kg⁻¹ soil compared to control. The application of S helped in reducing Cd toxicity, which was greater for 25 and 50 mg Cd kg⁻¹ soil) compared to 100 mg Cd kg⁻¹ soil. Addition of S to Cd-treated plants showed decrease in Cd and TBARS content in leaves and restoration of growth and photosynthesis through increase in the contents of AsA and GSH. Net photosynthetic rate and plant dry mass were strongly and positively correlated with the contents of AsA and GSH. It is suggested that S may ameliorate Cd toxicity and protects growth and photosynthesis of mustard involving AsA and GSH.     

Potential Phytoextraction and Phytostabilization of Perennial Peanut on Copper-Contaminated Vineyard Soils and Copper Mining Waste

This study sought to evaluate the potential of perennial peanut (Arachis pintoi) for copper phytoremediation in vineyard soils (Inceptisol and Mollisol) contaminated with copper and copper mining waste. Our results showed high phytomass production of perennial peanut in both vineyard soils. Macronutrient uptakes were not negatively affected by perennial peanut cultivated in all contaminated soils. Plants cultivated in Mollisol showed high copper concentrations in the roots and shoots of 475 and 52 mg kg⁻¹, respectively. Perennial peanut plants showed low translocation factor values for Cu, although these plants showed high bioaccumulation factor (BCF) for both vineyard soils, Inceptisol and Mollisol, with BCF values of 3.83 and 3.24, respectively, being characterized as a copper hyperaccumulator plant in these soils. Copper phytoextraction from Inceptisol soil was the highest for both roots and entire plant biomass, with more than 800 mg kg⁻¹ of copper in whole plant. The highest potential copper phytoextraction by perennial peanut was in Inceptisol soil with copper removal of 2,500 g ha⁻¹. Also, perennial peanut showed high potential for copper phytoremoval in copper mining waste and Mollisol with 1,700 and 1,500 g of copper per hectare, respectively. In addition, perennial peanuts characterized high potential for phytoextraction and phytostabilization of copper in vineyard soils and copper mining waste.     

Optimization and enhancement of soil bioremediation by composting using the experimental design technique

The objective of this study was the application of the experimental design technique to optimize the conditions for the bioremediation of contaminated soil by means of composting. A low-cost material such as compost from the Organic Fraction of Municipal Solid Waste as amendment and pyrene as model pollutant were used. The effect of three factors was considered: pollutant concentration (0.1–2 g/kg), soil:compost mixing ratio (1:0.5–1:2 w/w) and compost stability measured as respiration index (0.78, 2.69 and 4.52 mg O₂ g⁻¹ Organic Matter h⁻¹). Stable compost permitted to achieve an almost complete degradation of pyrene in a short time (10 days). Results indicated that compost stability is a key parameter to optimize PAHs biodegradation. A factor analysis indicated that the optimal conditions for bioremediation after 10, 20 and 30 days of process were (1.4, 0.78, 1:1.4), (1.4, 2.18. 1:1.3) and (1.3, 2.18, 1:1.3) for concentration (g/kg), compost stability (mg O₂ g⁻¹ Organic Matter h⁻¹) and soil:compost mixing ratio, respectively.     

Isolation and characterization of an arsenate-reducing bacterium and its application for arsenic extraction from contaminated soil

A Gram-negative anaerobic bacterium, Citrobacter sp. NC-1, was isolated from soil contaminated with arsenic at levels as high as 5,000 mg As kg⁻¹. Strain NC-1 completely reduced 20 mM arsenate within 24 h and exhibited arsenate-reducing activity at concentrations as high as 60 mM. These results indicate that strain NC-1 is superior to other dissimilatory arsenate-reducing bacteria with respect to arsenate reduction, particularly at high concentrations. Strain NC-1 was also able to effectively extract arsenic from contaminated soils via the reduction of solid-phase arsenate to arsenite, which is much less adsorptive than arsenate. To characterize the reductase systems in strain NC-1, arsenate and nitrate reduction activities were investigated using washed-cell suspensions and crude cell extracts from cells grown on arsenate or nitrate. These reductase activities were induced individually by the two electron acceptors. This may be advantageous during bioremediation processes in which both contaminants are present.     

Bioremediation of PAH-contaminated soil with fungi – From laboratory to field scale

The purpose of this study was to develop a fungal bioremediation method that could be used for soils heavily contaminated with persistent organic compounds, such as polyaromatic hydrocarbons (PAHs). Sawmill soil, contaminated with PAHs, was mixed with composted green waste (1:1) and incubated with or without fungal inoculum. The treatments were performed at the laboratory and field scales. In the laboratory scale treatment (starting concentration 3500 mg kg⁻¹, sum of 16 PAH) the high molecular weight PAHs were degraded significantly more in the fungal-inoculated microcosms than in the uninoculated ones. In the microcosms inoculated with Phanerochaete velutina, 96% of 4-ring PAHs and 39% of 5- and 6-ring PAHs were removed in three months. In the uninoculated microcosms, 55% of 4-ring PAHs and only 7% of 5- and 6-ring PAHs were degraded. However, during the field scale (2 t) experiment at lower starting concentration (1400 mg kg⁻¹, sum of 16 PAH) the % degradation was similar in both the P. velutina-inoculated and the uninoculated treatments: 94% of the 16 PAHs were degraded in three months. In the field scale experiment the copy number of gram-positive bacteria PAH-ring hydroxylating dioxygenase genes was found to increase 1000 fold, indicating that bacterial PAH degradation also played an important role.     

Accumulation and Speciation of Selenium in Plants as Affected by Arbuscular Mycorrhizal Fungus <Emphasis Type="BoldItalic">Glomus mosseae</Emphasis>

Effects of arbuscular mycorrhizal fungus (Glomus mosseae) on the accumulation and speciation of selenium (Se) in alfalfa, maize, and soybean were investigated by using Se(IV)-spiked soil. Mycorrhizal inoculation decreased Se accumulation in roots and shoots of all the plants at Se spiked level of 0 or 2 mg kg⁻¹, while an increased Se accumulation was observed in alfalfa shoots and maize roots and shoots at the spiked level of 20 mg kg⁻¹. Concentration of inorganic Se (especially Se(VI)) in roots and shoots of the three plants was much higher in mycorrhizal than non-mycorrhizal treatment. Mycorrhizal inoculation decreased the portion of total organic Se in plant tissues with the exception of alfalfa and maize shoots at Se spiked level of 20 mg kg⁻¹, in which organic Se portion did not reduced greatly (<5%) for mycorrhizal treatment. Mycorrhizal effects on alfalfa and maize were more obvious than on soybean in terms of root colonization rate, biomass, and Se accumulation.