Evaluation of chromium(VI) removal behaviour by two isolates of Synechocystis sp. in terms of exopolysaccharide (EPS) production and monomer composition

Chromium(VI) removal and its association with exopolysaccharide (EPS) production in cyanobacteria were investigated. Synechocystis sp. BASO670 produced higher EPS (548 mg L⁻¹) than Synechocystis sp. BASO672 (356 mg L⁻¹). While the EC₅₀ of the Cr(VI) for Synechocystis sp. BASO670 and Synechocystis sp. BASO672 were determined as 11.5 mg L⁻¹, and 2.0 mg L⁻¹, respectively, there was no relation between Cr(VI) removal and EPS production. Synechocystis sp. BASO672, which has higher EPS value, removed (33%) more Cr(VI) than Synechocystis sp. BASO670. Monomer compositions of EPS of each of the isolates were determined differently. Synechocystis sp. BASO672 which removed higher Cr(VI), had higher values of uronic acid and glucuronic acid (192 μg/mg and 89%, respectively). Our results showed that EPS might play a role in Cr(VI) tolerance. Monomer composition, especially uronic acid and glucuronic acid content of EPS may have enhanced Cr(VI) removal.     

Distinct and effective biotransformation of hexavalent chromium by a novel isolate under aerobic growth followed by facultative anaerobic incubation

A bacterial isolate (G161) with high Cr(VI)-reducing capacity was isolated from Cr(VI)-contaminated soil and identified as Leucobacter sp. on the basis of 16S rRNA gene sequence analysis. The isolate was a Gram-positive, aerobic rod. The hexavalent chromate-reducing capability of the isolate was investigated under three conditions of oxygen stress. The isolate was found to reduce Cr(VI) under all conditions but performed most effectively during aerobic growth followed by facultative anaerobic incubation. Under these conditions, the isolate tolerated K₂Cr₂O₇ concentrations up to 1,000 mg/l and completely reduced 400 mg/l K₂Cr₂O₇ within 96 h. The strain reduced Cr(VI) over a wide range of pH (6.0–11.0) and temperatures (15–45 °C) with optimum performance at pH?8.0 and 35 °C. The presence of other metals, such as Ca²⁺, Co²⁺, Cu²⁺, Mn²⁺, Ni²⁺, and Zn²⁺, induced no effect or else played a stimulatory role on Cr(VI)-reduction activity of the strain. The strain was tested for Cr(VI) removal in wastewaters and proved capable of completely reducing the contained Cr(VI). This is the novel report of a bacterial growth and Cr(VI)-reduction process under sequential aerobic growth and facultative anaerobic conditions. The study suggested that the isolate possesses a distinct capability for Cr(VI) reduction which could be harnessed for the detoxification of chromate-contaminated wastewaters.     

Medium optimization for the production of a novel bioflocculant from Halomonas sp. V3a′ using response surface methodology

The novel exopolysaccharide bioflocculant HBF-3 is produced by Halomonas sp. V3a′, which is a mutant strain of the deep-sea bacterium Halomonas sp. V3a. Response surface methodology (RSM) was employed to optimize the production medium for increasing HBF-3 production. Using a Plackett–Burman experimental design to aid in the first step of optimization, edible glucose, MgSO₄·7H₂O, and NH₄Cl were found to be significant factors affecting HBF-3 production. To determine the optimal concentration of each significant variable, a central composite design was employed. Based on response surface and canonical analysis, the optimum concentrations of the critical components were obtained as follows: edible glucose, 16.14 g/l; MgSO₄·7H₂O, 2.73 g/l; and NH₄Cl, 1.97 g/l. HBF-3 production obtained by using the optimized medium was 4.52 g/l, which was in close agreement with the predicted value of 4.55 g/l. By scaling up fermentation from flask to fermenter, HBF-3 production was further increased to 5.58 g/l.     

Extraction and optimization of exopolysaccharide from Lactobacillus sp. using response surface methodology and artificial neural networks

Abstract

The microbial polysaccharides secreted and produced from various microbes into their extracellular environment is known as exopolysaccharide. These polysaccharides can be secreted from the microbes either in a soluble or insoluble form.Lactobacillus sp. is one of the organisms that have been found to produce exopolysaccharide. Exo-polysaccharides (EPS) have various applications such as drug delivery, antimicrobial activity, surgical implants and many more in different fields. Medium composition is one of the major aspects for the production of EPS from Lactobacillus sp., optimization of medium components can help to enhance the synthesis of EPS . In the present work, the production of exopolysaccharide with different medium composition was optimized by response surface methodology (RSM) followed by tested for fitting with artificial neural networks (ANN). Three algorithms of ANN were compared to investigate the highest yeild of EPS. The highest yeild of EPS production in RSM was achieved by the medium composition that consists of (g/L) dextrose 15, sodium dihydrogen phosphate 3, potassium dihydrogen phosphate 2.5, triammonium citrate 1.5, and, magnesium sulfate 0.25. The output of 32 sets of RSM experiments were tested for fitting with ANN with three algorithms viz. Levenberg–Marquardt Algorithm (LMA), Bayesian Regularization Algorithm (BRA) and Scaled Conjugate Gradient Algorithm (SCGA) among them LMA found to have best fit with the experiments as compared to the SCGA and BRA.     

Biosorption of Cr(VI) by Ceratocystis paradoxa MSR2 Using Isotherm Modelling,Kinetic Study and Optimization of Batch Parameters Using Response Surface Methodology

This study is focused on the possible use of Ceratocystis paradoxa MSR2 native biomass for Cr(VI) biosorption. The influence of experimental parameters such as initial pH, temperature, biomass dosage, initial Cr(VI) concentration and contact time were optimized using batch systems as well as response surface methodology (RSM). Maximum Cr(VI) removal of 68.72% was achieved, at an optimal condition of biomass dosage 2g L⁻¹, initial Cr(VI) concentration of 62.5 mg L⁻¹ and contact time of 60 min. The closeness of the experimental and the predicted values exhibit the success of RSM. The biosorption mechanism of MSR2 biosorbent was well described by Langmuir isotherm and a pseudo second order kinetic model, with a high regression coefficient. The thermodynamic study also revealed the spontaneity and exothermic nature of the process. The surface characterization using FT-IR analysis revealed the involvement of amine, carbonyl and carboxyl groups in the biosorption process. Additionally, desorption efficiency of 92% was found with 0.1 M HNO₃. The Cr(VI) removal efficiency, increased with increase in metal ion concentration, biomass concentration, temperature but with a decrease in pH. The size of the MSR2 biosorbent material was found to be 80 μm using particle size analyzer. Atomic force microscopy (AFM) visualizes the distribution of Cr(VI) on the biosorbent binding sites with alterations in the MSR2 surface structure. The SEM-EDAX analysis was also used to evaluate the binding characteristics of MSR2 strain with Cr(VI) metals. The mechanism of Cr(VI) removal of MSR2 biomass has also been proposed.     

Biosorption of chromium(VI) by spent cyanobacterial biomass from a hydrogen fermentor using Box-Behnken model

The study explores utilization of waste cyanobacterial biomass of Nostoc linckia from a lab-scale hydrogen fermentor for the biosorption of Cr(VI) from aqueous solution. The biomass immobilized in alginate beads was used for removal of the metal in batch mode optimizing the process conditions adopting response surface methodology (RSM). Kinetic studies were done to get useful information on the rate of chromium adsorption onto the cyanobacterial biomass, which was found to follow pseudo second-order model. Four important process parameters including initial metal concentration (10-100 mg/L), pH (2-6), temperature (25-45 °C) and cyanobacterial dose (0.1-2.0 g) were optimized to obtain the best response of Cr(VI) removal using the statistical Box-Behnken design. The response surface data indicated maximum Cr(VI) biosorption at pH 2-4 with different initial concentrations of the metal in the aqueous solution. The biosorbent could remove 80-90% chromium from solutions with initial metal concentration of 10-55 mg/L. Involvement of the surface characteristics of the biomass was studied through its scanning electron micrographs and Fourier transform infrared (FTIR) analysis.     

Simultaneous bioremediation of Cr(VI) and lindane in soil by actinobacteria

Environments co-contaminated with metals and organic compounds are difficult to remediate. Actinobacteria is an important group of microorganisms found in soils, with high metabolic versatility and potential for bioremediation. In this paper, actinobacteria were used to remediate soil co-contaminated with Cr(VI) and lindane. Five actinobacteria, tolerant to Cr(VI) and lindane mixture were selected: Streptomyces spp. A5, A11, M7, and MC1, and Amycolatopsis tucumanensis DSM 45259. Sterilized soil samples were inoculated with actinobacteria strains, either individually or as a consortium, and contaminated with Cr(VI) and lindane, either immediately or after 7 days of growth, and incubated at 30 °C during 14 days. All actinobacteria were able to grow and remove both contaminants, the consortium formed by Streptomyces spp. A5, M7, MC1, and A. tucumanensis showed the highest Cr(VI) removal, while Streptomyces sp. M7 produced the maximum lindane removal. In non-sterile soil samples, Streptomyces sp. M7 and the consortium removed more than 40% of the lindane, while Streptomyces sp. M7 demonstrated the greatest Cr(VI) removal. The most appropriate strategy for bioremediation of Cr(VI) and lindane co-contaminated soils would be the inoculation with Streptomyces sp. M7.     

Production,Purification, and Antibiofilm Activity of a Novel Exopolysaccharide from Arthrobacter sp. B4

A novel exopolysaccharide (EPS), namely, B4-EPS, is produced by Arthrobacter sp. B4. Response surface methodology (RSM) was employed to optimize the fermentation medium for increasing B4-EPS production. Based on Plackett–Burman design (PBD), glucose, yeast extract, and KH₂PO₄ were selected as significant variables, which were further optimized by a central composite design (CCD). According to response surface and canonical analysis, the optimal medium was composed of 16.94 g/L glucose, 2.33 g/L yeast extract, and 5.32 g/L KH₂PO₄. Under this condition, the maximum yield of B4-EPS reached about 8.54 g/L after 72 hr of batch fermentation, which was pretty close to the predicted value (8.52 g/L). Furthermore, B4-EPS was refined by column chromatography. The main homogeneous fraction (B4-EPS1) was collected and applied to assay of antibiofilm activity. B4-EPS1 exhibited a dose-dependent inhibitory effect on biofilm formation of Pseudomonas aeruginosa PAO1 without antibacterial activity. About 86.1% of biofilm formation of P. aeruginosa PAO1 was inhibited in the presence of 50 µg/mL B4-EPS1, which was more effective than the peer published data. Moreover, B4-EPS1 could prevent biofilm formation of other strains. These data suggest B4-EPS may represent a promising strategy to combat bacterial biofilms in the future.     

Isolation,identification and characterization of a novel Rhodococcus sp. strain in biodegradation of tetrahydrofuran and its medium optimization using sequential statistics-based experimental designs

Statistics-based experimental designs were applied to optimize the culture conditions for tetrahydrofuran (THF) degradation by a newly isolated Rhodococcus sp. YYL that tolerates high THF concentrations. Single factor experiments were undertaken for determining the optimum range of each of four factors (initial pH and concentrations of K₂HPO₄ · 3H₂O, NH₄Cl and yeast extract) and these factors were subsequently optimized using the response surface methodology. The Plackett–Burman design was used to identify three trace elements (Mg²⁺, Zn²⁺and Fe²⁺) that significantly increased the THF degradation rate. The optimum conditions were found to be: 1.80 g/L NH₄Cl, 0.81 g/L K₂HPO₄ · 3H₂O, 0.06 g/L yeast extract, 0.40 g/L MgSO₄ · 7H₂O, 0.006 g/L ZnSO₄ · 7H₂O, 0.024 g/L FeSO₄ · 7H₂O, and an initial pH of 8.26. Under these optimized conditions, the maximum THF degradation rate increased to 137.60 mg THF h⁻¹ g dry weight in Rhodococcus sp. YYL, which was nearly five times of that by the previously described THF degrading Rhodococcus strain.     

Investigation of Cr(VI) adsorption onto chemically treated Helianthus annuus: Optimization using Response Surface Methodology

In the present study, chemically treated Helianthus annuus flowers (SHC) were used to optimize the removal efficiency for Cr(VI) by applying Response Surface Methodological approach. The surface structure of SHC was analyzed by Scanning Electron Microscopy (SEM) coupled with Energy Dispersive X-ray Analysis (EDX). Batch mode experiments were also carried out to assess the adsorption equilibrium in aqueous solution. The adsorption capacity (q_e) was found to be 7.2 mg/g. The effect of three parameters, that is pH of the solution (2.0-7.0), initial concentration (10-70 mg/L) and adsorbent dose (0.05-0.5 g/100 mL) was studied for the removal of Cr(VI) by SHC. Box-Behnken model was used as an experimental design. The optimum pH, adsorbent dose and initial Cr(VI) concentration were found to be 2.0, 5.0 g/L and 40 mg/L, respectively. Under these conditions, removal efficiency of Cr(VI) was found to be 90.8%.     

Production of 2,3-Butanediol from Sucrose by a <Emphasis Type="Italic">Klebsiella</Emphasis> Species

Chemical 2,3-butanediol is an important platform compound possessing diverse industrial applications. So far, it is mainly produced by using petrochemical feedstock which is associated with high cost and adverse environmental impacts. Hence, finding alternative routes (e.g., via fermentation using renewable carbon sources) to produce 2,3-butanediol are urgently needed. In this study, we report a wild-type Klebsiella sp. strain XRM21, which is capable of producing 2,3-butanediol from a wide variety of carbon sources including glucose, sucrose, xylose, and glycerol. Among them, fermentation of sucrose leads to the highest production of 2,3-butanediol. To maximize the production of 2,3-butanediol, fermentation conditions were first optimized for strain XMR21 by using response surface methodology (RSM) in batch reactors. Subsequently, a fed-batch fermentation strategy was designed based on the optimized parameters, where 91.2 g/L of 2,3-butanediol could be produced from substrate sucrose dosing in 100 g/L for three times. Moreover, random mutagenesis of stain XMR21 resulted in a highly productive mutant strain, capable of producing 119.4 and 22.5 g/L of 2,3-butanediol and ethanol under optimized fed-batch fermentation process within 65 h with a total productivity of 2.18 g/L/h, which is comparable to the reported highest 2,3-butanediol concentration produced by previous strains. This study provides a potential strategy to produce industrially important 2,3-butanediol from low-cost sucrose.     

Cr(Ⅵ)还原菌Microbacterium sp.BD6的分离鉴定及还原特性

【目的】从电镀厂下水道的淤泥中分离筛选Cr(Ⅵ)高效还原菌,并对其生长和还原特性进行研究,以期为Cr(Ⅵ)污染的生物修复提供优质的菌种资源和应用参考。【方法】采用富集培养法从淤泥中分离、筛选出Cr(Ⅵ)还原菌,通过生理生化及16S rRNA基因序列分析进行初步鉴定。采用单因素实验确定菌株的最佳培养条件和抵抗胁迫环境的能力,利用外加电子供体改善菌株的Cr(Ⅵ)还原能力,筛选出最佳电子供体研究对菌株还原的影响。【结果】经分离筛选得到1株Cr(Ⅵ)耐受还原菌,初步鉴定为微杆菌属(Microbacterium sp.),命名为BD6。菌株BD6适宜在中温、偏碱性的环境条件下生长,能耐受50.0 g/L NaCl的高盐环境。Mn^2+对菌种的生长表现出较高的抑制,Ni^2+、Zn^2+、Cd^2+的抑制作用较小,Cu^2)产生了一定的促进作用。Cr(Ⅵ)对BD6的最低抑菌浓度为1700 mg/L。添加甘油、果糖、乳糖、葡萄糖、丙酮酸钠作为电子供体促进了菌株对Cr(Ⅵ)的还原。选择甘油作为菌株还原Cr(Ⅵ)的最佳电子供体,无电子供体添加时菌株96 h内对100 mg/L Cr(Ⅵ)的还原率仅为69.63%,添加2 g/L的甘油菌株在36 h内的还原率达到了100%。通过加大甘油的添加量可以促进菌株对初始浓度较高Cr(Ⅵ)的还原,但要受到Cr(Ⅵ)的毒性限制。菌株的最适还原条件和最适生长条件吻合,在50.0 g/L NaCl的高盐条件和50 mg/L Cd^2+的毒性环境中,添加2 g/L的甘油,菌株对100 mg/L Cr(Ⅵ)的还原率分别为72 h 96.79%、54 h 99.86%。【结论】分离筛选得到的Microbacterium sp.BD6是一株潜在的可用于Cr(Ⅵ)污染生物还原修复的候选菌株。     

Isolation and characterization of chromium(VI)-reducing bacteria from tannery effluents and solid wastes

In the present investigation, five novel Cr(VI) reducing bacteria were isolated from tannery effluents and solid wastes and identified as Kosakonia cowanii MKPF2, Klebsiella pneumonia MKPF5, Acinetobacter gerneri MKPF7, Klebsiella variicola MKPF8 and Serratia marcescens MKPF12 by 16S rDNA gene sequence analysis. The maximum tolerance concentration of Cr(VI) as K₂Cr₂O₇ of the bacterial isolates was varying up to 2000 mg/L. Among the investigated bacterial isolates, A. gerneri MKPF7 was best in terms of reduction rate. The optimum temperatures for growth and Cr(VI) reduction by the bacterial isolates were 35 and 40 °C, respectively except A. gerneri MKPF7 which grew and reduced Cr(VI) optimally at 40 °C. The optimum pH for growth and Cr(VI) reduction by K. cowanii MKPF2, A. gerneri MKPF7 and S. marcescens MKPF12 was 7.0 whereas the optimum pH for growth and Cr(VI) reduction by K. pneumoniae MKPF5 and K. variicola MKPF8 were 7.0, 8.0 and 6.0, 7.0, respectively. All the bacterial isolates showed maximum tolerance against Ni²⁺ and Zn²⁺ whereas minimum tolerance was observed against Hg²⁺ and Cd²⁺. The bacteria isolated in the present study thus can be used as eco-friendly biological expedients for the remediation and detoxification of Cr(VI) from the contaminated environments.     

Bioreduction of Cr(VI) by Bacillus sp. QH-1 isolated from soil under chromium-containing slag heap in high altitude area

A chromium-reducing strain QH-1, identified as Bacillus sp., was isolated from soil under chromium-containing slag heap in Qinghai high altitude area, China. The strain was found to resist 200 mg/L Cr(VI), and Cr(VI) negatively affects the metabolic activity of the cells, as well as the cell morphology of Bacillus sp. QH-1. The reduction efficiency of Cr(VI) at concentrations of Cr(VI) 25 mg/L, 50 mg/L, 100 mg/L and 200 mg/L were 99.48 %, 65.99 %, 23.22 % and 6.99 %, respectively, decreasing with increasing initial Cr(VI) concentration. This indicates that the toxicity of Cr(VI) increased with concentration. Energy dispersive X-ray analysis revealed that there was insoluble Cr(III) generated during Cr(VI) reduction. In order to apply strain QH-1 to remove Cr(VI) from groundwater, factors of concentration of electron donors (glucose) and temperature were investigated in a synthetic medium. The results demonstrated that glucose could promote the reduction of Cr(VI) by this strain, and the general trend of Cr(VI) reduction increased with temperature within the range of 4 to 37 °C. Cr(VI) was reduced effectively at 25 °C and 37 °C, and all of Cr(VI) was reduced after 96 h at 37 °C, while the reduction was slow at 4 °C and 15 °C, and it almost ceased after about 120 h. These results could be potentially useful for the bioremediation of Cr(VI) in groundwater.     

Production and characterization of curdlan by Agrobacterium sp.

Agrobacterium sp. was studied for the production of curdlan by conventional one-factor-at-a-time technique and response surface methodology. Factors such as initial pH, urea concentration, sucrose concentration having the greatest influence on the curdlan production were identified. By using response surface methodology (RSM), the curdlan production by Agrobacterium sp. was increased significantly by 109%, from 2.4 g/L to 5.02 g/L when the strain was cultivated in the optimal medium developed by RSM as compared to conventional one-factor-at-a-time technique. The curdlan production rate of 0.84 g/(L h) was obtained when Agrobacterium sp. was cultivated in the optimal medium developed by RSM, which was the highest curdlan production rate reported to date. The infrared (IR) and NMR spectra, the thermogram of DSC and pattern of X-ray diffraction for the curdlan of the present study were almost identical to those of the authentic curdlan sample (from Alcaligenes faecalis; Sigma). The purified curdlan was a linear polysaccharide composed of exclusively β-(1,3)-glucosidic linkages with the molecular weight of 160,000 Da by GPC. The crystalline melting point (T_m), glass transition temperature (T_g) and X-ray diffraction of the sample indicated low crystallinity in the structure.     

Evaluation and elimination of inhibitory effects of salts and heavy metal ions on biodegradation of Congo red by Pseudomonas sp. mutant

In this study, it was attempted to evaluate the influences and also recommended some elimination methods for inhibitory effects offered by salts and heavy metal ions. Congo red dye solution treated with mutant Pseudomonas sp. was taken as a model system for study. The salts used in this study are NaCl, CaCl₂ and MgSO₄·7H₂O. Though the growth was inhibited at concentrations above 4 g/l, toleration was achieved by acclimatization process. In case of heavy metal ions, Cr (VI) showed low inhibition up to 500 mg/l of concentration, compared to Zn (II) and Cu (II). It was due to the presence of chromium reductase enzyme which was confirmed by SDS-PAGE. Zn (II) and Cu (II) ion inhibitions were eliminated by chelation with EDTA. The critical ion concentrations obtained as per Han-Levenspiel model for Cr (VI), Zn (II) and Cu (II) were 0.8958, 0.3028 and 0.204 g/l respectively.     

Toxicity of Hexavalent Chromium and Its Reduction by Bacteria Isolated from Soil Contaminated with Tannery Waste

An Arthrobacter sp. and a Bacillus sp., isolated from a long-term tannery waste contaminated soil, were examined for their tolerance to hexavalent chromium [Cr(VI)] and their ability to reduce Cr(VI) to Cr(III), a detoxification process in cell suspensions and cell extracts. Both bacteria tolerated Cr(VI) at 100 mg/ml on a minimal salts agar medium supplemented with 0.5% glucose, but only Arthrobacter could grow in liquid medium at this concentration. Arthrobacter sp. could reduce Cr(VI) up to 50 μg/ml, while Bacillus sp. was not able to reduce Cr(VI) beyond 20 μg/ml. Arthrobacter sp. was distinctly superior to the Bacillus sp. in terms of their Cr(VI)-reducing ability and resistance to Cr(VI). Assays with permeabilized (treated with toluene or Triton X 100) cells and crude extracts demonstrated that the Cr(VI) reduction was mainly associated with the soluble protein fraction of the cell. Arthrobacter sp. has a great potential for bioremediation of Cr(VI)-containing waste. Received: 13 June 2002 / Accepted: 13 September 2002     

Cr(VI) removal from aqueous solution by thermophilic denitrifying bacterium <Emphasis Type="Italic">Chelatococcus daeguensis</Emphasis> TAD1 in the presence of single and multiple heavy metals

Cr(VI) pollution is increasing continuously as a result of ongoing industrialization. In this study, we investigated the thermophilic denitrifying bacterium Chelatococcus daeguensis TAD1, isolated from the biofilm of a biotrickling filter used in nitrogen oxides (NO_X) removal, with respect to its ability to remove Cr(VI) from an aqueous solution. TAD1 was capable of reducing Cr(VI) from an initial concentration of 10 mg/L to non-detectable levels over a pH range of 7–9 and at a temperature range of 30–50°C. TAD1 simultaneously removed both Cr(VI) and NO₃^?-N at 50°C, when the pH was 7 and the initial Cr(VI) concentration was 15 mg/L. The reduction of Cr(VI) to Cr(III) correlated with the growth metabolic activity of TAD1. The presence of other heavy metals (Cu, Zn, and Ni) inhibited the ability of TAD1 to remove Cr(VI). The metals each individually inhibited Cr(VI) removal, and the extent of inhibition increased in a cooperative manner in the presence of a combination of the metals. The addition of biodegradable cellulose acetate microspheres (an adsorption material) weakened the toxicity of the heavy metals; in their presence, the Cr(VI) removal efficiency returned to a high level. The feasibility and applicability of simultaneous nitrate removal and Cr(VI) reduction by strain TAD1 is promising, and may be an effective biological method for the clean-up of wastewater.     

Distribution and speciation of chromium accumulated in Gynura pseudochina (L.) DC.

Soil and water contamination with chromium is an issue of recent concern in Thailand due to increases in industrial activity. Gynura pseudochina (L.) DC., a chromium tolerance plant, could be employed to address this problem via phytoremediation. To understand the tolerance mechanism, this study investigated the speciation and distribution of chromium accumulated in G. pseudochina (L.) DC. using AAS, XAFS, μ-XANES, μ-XRF imaging and EPR. The plants were separately treated with K₂Cr₂O₇ and Cr₂(SO₄)₃ in a hydroponic system. μ-XRF imaging clarified the distributions of Cr, Fe, Zn, Ca, Cl, K and S within the samples. In G. pseudochina (L.) DC. treated with Cr(VI) solution, the Cr was mainly distributed in the vascular bundle and periderm of the tuber, the stem xylem, the vein and the epidermis, including the trichome of the leaf tissues. This Cr distribution corresponded to those of Cu, Fe and Zn. In G. pseudochina (L.) DC. treated with Cr(III) solution, the Cr was distributed in the periderm of the tuber, the stem cortex, and the epidermis and parenchyma of the leaf tissues. μ-XANES and XAFS indicated that highly toxic Cr(VI) was reduced to the intermediate Cr(V) and accumulated as less toxic Cr(III), and EXAFS spectra showed that the reduced Cr(III) was bound to oxygen ligands. The coordination number (N) and the interatomic distance (R) to the first shell were approximately 3–4 (N) and 2 Å (R), respectively. EPR spectra of the plant samples treated with Cr(VI) revealed the presence of Cr(V) and Cr(III). Thus, Cr(III) and Cr(VI) were taken up into the vascular system and transported from the roots to the leaves. Cr(III) was distributed via the symplast system to the ground tissue and accumulated mainly in the stem cortex. Cr(VI) was transported to the xylem via the apoplast system, and the adsorption of Cr(VI) and its reduction to Cr(V) and Cr(III) occurred on oxygen ligands in the lignocellulosic structure of the xylem and vein.     

Biotoxic Effect of Chromium (VI) on Plant Growth-Promoting Traits of Novel Cellulosimicrobium funkei Strain AR8 Isolated from Phaseolus vulgaris Rhizosphere

The aim of this study was to investigate the effect of Cr(VI) on the plant growth-promoting traits of potential rhizobacterial strain isolated from Phaseolus vulgaris rhizosphere. A total of 36 rhizobacterial strains were recovered from the rhizosphere of P. vulgaris. Among these strains, the strain AR8 was specifically selected due to the highest resistance against heavy metals and the maximum production of plant growth-promoting substances. The rhizobacterial strain AR8 was identified as Cellulosimicrobium funkei (KM263188) following 16S rDNA gene sequencing. Strain AR8 solubilized phosphate and produced indole-3-acetic acid (32.57 µg/ml), exopolysaccharide (17.23 µg/ml), ammonia (54.16 µg/ml), catalase, biosurfactant, protease, amylase, and lipase. Under Cr(VI) stress, Cr(VI) concentration-dependent progressive decline in all plant growth-promoting traits of the C. funkei exposed was observed except for exopolysaccharide production, which consistently increased with increasing concentrations of Cr(VI). The root elongation assay resulted that the application of C. funkei strain AR8 significantly increased root length of test crops both in the presence and absence of Cr(VI) compared to uninoculated Cr(VI) treated plants. Moreover, AR8 generated a large number of colonies in diverse agricultural crops. Due to these intrinsic abilities, strain AR8 could be utilized for growth promotion as well as for the remediation of chromium in chromium-contaminated soil.