Significance of Bacillus subtilis strain SJ-101 as a bioinoculant for concurrent plant growth promotion and nickel accumulation in Brassica juncea

In this study, a nickel (Ni)-tolerant Bacillus subtilis strain SJ-101 was characterized based on the 16SrDNA homology and phylogenetic analysis. The role of this strain ascertained in facilitating Ni accumulation in the Indian mustard plant (Brassica juncea [L]. Czern and Coss) var. Pusa Bold (DIR-50), to elucidate the potential of Ni phytoremediation in combination with metal-tolerant rhizobacteria. The data revealed that the plants exposed to NiCl2 (1750 mg kg(-1)) in soil bioaugmented with strain SJ-101 have accumulated 0.147% Ni vis-à-vis 0.094% accumulation in dry biomass of the plants grown in uninoculated soil. The strain SJ-101 has also exhibited the capability of producing indole acetic acid (IAA) (55 microg ml(-1)), and solubilizing inorganic phosphate (90 microg ml(-1)) in specific culture media. The pot culture experiments clearly demonstrated the beneficial effects of bioinoculant strain SJ-101 with significant increase (p<0.05) in the plant growth attributes in untreated control soil. Furthermore, the protective effect of the strain SJ-101 against Ni phytotoxicity was evident in plants grown in soil treated with NiCl2 in concentration range of 250-1750 mg kg(-1). Thus, it is suggested that the strain SJ-101 owing to its intrinsic abilities of plant growth promotion, and attenuation of soil Ni by biosorption and bioaccumulation, could be exploited for bacteria-assisted phytoaccumulation of this toxic heavy metal from contaminated sites.     

Uptake of heavy metals and As by Brassica juncea grown in a contaminated soil in Aznalcóllar (Spain): the effect of soil amendments

Two crops of Brassica juncea (L.) Czern. were grown in a field experiment, at the site affected by the toxic spillage of acidic, metal-rich waste in Aznalcóllar (Seville, Spain), to study its metal accumulation and the feasibility of its use for metal phytoextraction. The effects of organic soil amendments (cow manure and mature compost) and lime on biomass production and plant survival were also assessed; plots without organic amendment and without lime were used as controls. Plots, with or without organic amendment, having pH < 5 were limed for the second crop. Soil acidification conditioned plant growth and metal accumulation. The addition of lime and the organic amendments achieved higher plant biomass production, although effects concerning metal bioavailability and accumulation were masked somewhat by pH variability with time and between and within plots. Tissue metal concentrations of B. juncea were elevated for Zn, Cu and Pb, especially in leaves of plants from plots with low pH values (maxima of 2029, 71 and 55 microg g(-1), respectively). The total uptake of heavy metals in the plants was relatively low, emphasising the problems faced when attempting to employ phytoextraction for clean-up of pluri-contaminated sites.     

Influence of plant growth promoting bacteria and Cr6+ on the growth of Indian mustard

The Cr(6+) resistant plant growth promoting bacteria (PGPB), Pseudomonas sp. PsA4 and Bacillus sp. Ba32 were isolated from heavy metal contaminated soils and their plant growth promoting activity on the Indian mustard (Brassica juncea) were assessed with different concentrations of Cr(6+) in soil. Production of siderophores and the solubilization of phosphate were observed in both strains, PsA4 and Ba32. Production of IAA was only observed in strain PsA4. Inoculation of PsA4 or Ba32 promoted the growth of plants at 95.3 and 198.3 microg of Cr(6+)g(-1) soil. The maximum growth was observed in the plants inoculated with strain PsA4. Both strains, PsA4 and Ba32 did not influence the quantity of accumulation of chromium in root and shoot system. The present observations showed that the strains PsA4 and Ba32 protect the plants against the inhibitory effects of chromium, probably due to the production of IAA, siderophores and solubilization of phosphate.     

The effects of soil amendments on heavy metal bioavailability in two contaminated Mediterranean soils

Two heavy metal contaminated calcareous soils from the Mediterranean region of Spain were studied. One soil, from the province of Murcia, was characterised by very high total levels of Pb (1572 mg kg(-1)) and Zn (2602 mg kg(-1)), whilst the second, from Valencia, had elevated concentrations of Cu (72 mg kg(-1)) and Pb (190 mg kg(-1)). The effects of two contrasting organic amendments (fresh manure and mature compost) and the chelate ethylenediaminetetraacetic acid (EDTA) on soil fractionation of Cu, Fe, Mn, Pb and Zn, their uptake by plants and plant growth were determined. For Murcia soil, Brassica juncea (L.) Czern. was grown first, followed by radish (Raphanus sativus L.). For Valencia soil, Beta maritima L. was followed by radish. Bioavailability of metals was expressed in terms of concentrations extractable with 0.1 M CaCl2 or diethylenetriaminepentaacetic acid (DTPA). In the Murcia soil, heavy metal bioavailability was decreased more greatly by manure than by the highly-humified compost. EDTA (2 mmol kg(-1) soil) had only a limited effect on metal uptake by plants. The metal-solubilising effect of EDTA was shorter-lived in the less contaminated, more highly calcareous Valencia soil. When correlation coefficients were calculated for plant tissue and bioavailable metals, the clearest relationships were for Beta maritima and radish.     

Combined application of Triton X-100 and Sinorhizobium sp. Pb002 inoculum for the improvement of lead phytoextraction by Brassica juncea in EDTA amended soil

The process of EDTA-assisted lead phytoextraction from the Bovisa (Milan, Italy) brownfield soil was optimized in microcosms vegetated with Brassica juncea. An autochthonous plant growth-promoting rhizobacterium (PGPR), Sinorhizobium sp. Pb002, was isolated from the rhizosphere of B. juncea grown on the Pb-contaminated soil in presence of 2 mM EDTA. The strain was augmented (10(8) CFU g(-1) soil) in vegetated microcosms to stimulate B. juncea biomass production and, hence, its phytoextraction potential. Triton X-100 was also added to microcosms at 5 and 10 times the critical micelle concentration (cmc) to increase the permeability of root barriers to the EDTA-Pb complexes. Triton X-100 amendment determined an increase in Pb concentration within plant tissues. However it contextually exerted a phytotoxic effect. Sinorhizobium sp. Pb002 augmentation was crucial to plant survival in presence of both bioavailable lead and Triton X-100. The combination of the two treatments produced up to 56% increase in the efficiency of lead phytoextraction by B. juncea. The effects of these treatments on the structure of the soil bacterial community were evaluated by 16S rDNA denaturing gradient gel electrophoresis (DGGE).     

Phytoextraction of metals from a multiply contaminated soil by Indian mustard

The effects of nitrilotriacetate (NTA) and citric acid applications on metal extractability from a multiply metal-contaminated soil, as well as on their uptake and accumulation by Indian mustard (Brassica juncea) were investigated. Desorption of metals from the soil increased with chelate concentration, NTA being more effective than citric acid in solubilising the metals. Plants were grown in a sandy soil collected from a contaminated field site and polluted by Cd, Cr, Cu, Pb and Zn. After 43 days of plant growth, pots were amended with NTA or citric acid at 5 mmol kg-1 soil. Control pots were not treated with any chelate. Harvest of plants was performed 1 week after chelate addition. Soil water-, NH4NO3- and DTPA-extractable Cd, Cu, Pb and Zn fractions were enhanced only in the presence of NTA. In comparison to unamended plants, Indian mustard shoot dry weights suffered significant reductions following NTA application. NTA treatment increased shoot metal concentrations by a factor of 2-3, whereas citric acid did not induce any difference compared to the control. Chromium was detected in the above-ground tissues only after NTA amendment. Due to differences in dry matter yield, a significant enhancement of metal uptake was observed in NTA-treated plants for Cu and Zn.     

Trichoderma atroviride F6 improves phytoextraction efficiency of mustard (Brassica juncea (L.) Coss. var. foliosa Bailey) in Cd, Ni contaminated soils

Trichoderma atroviride F6, isolated from decaying feather and resistant to 100 mg l(-1) Cd2+ and 250 mg l(-1) Ni2+, was applied for rhizoremediation of Cd, Ni and Cd-Ni combination contaminated soils through association with Brassica juncea (L.) Coss. var. foliosa. The strain significantly alleviated the cellular toxicity of cadmium and nickel to plants. Inoculation of B. juncea (L.) Coss. var. foliosa with T. atroviride F6 resulted a 110%, 40% and 170% increase in fresh weight in Cd, Ni and Cd-Ni contaminated soils, respectively (P<0.05). The translocation factors and metal bioconcentration factors calculated for the inoculated plant were increased compared to the noninoculated plants. The results indicated that the efficiency of phytoextraction for B. juncea (L.) Coss. var. foliosa enhanced after inoculating with T. atroviride F6. The fungal treated plants grown in Cd-Ni combination contaminated soils showed higher phytoextraction efficiency than those in Cd or Ni contaminated soils. Thus, it is suggested that the fungus T. atroviride F6 endowed with organic-degrading capabilities could be exploited for fungi-assisted phytoremediation of mixed organic-metal contaminated soils.     

Growth performance and biochemical responses of three rice (Oryza sativa L.) cultivars grown in fly-ash amended soil

The disposal of fly-ash (FA) from coal-fired power stations causes significant economic and environmental problems. Use of such contaminated sites for crop production and use of contaminated water for irrigation not only decreases crop productivity but also poses health hazards to humans due to accumulation of toxic metals in edible grains. In the present investigation, three rice cultivars viz., Saryu-52, Sabha-5204, and Pant-4 were grown in garden soil (GS, control) and various amendments (10%, 25%, 50%, 75% and 100%) of FA for a period of 90 days and effect on growth and productivity of plant was evaluated vis-a-vis metal accumulation in the plants. The toxicity of FA at higher concentration (50%) was reflected by the reduction in photosynthetic pigments, protein and growth parameters viz., plant height, root biomass, number of tillers, grain and straw weight. However, at lower concentrations (10-25%), FA enhanced growth of the plants as evident by the increase of studied growth parameters. The cysteine and non-protein thiol (NP-SH) content showed increase in their levels up to 100% FA as compared to control, however, maximum content was found at 25% FA in Saryu-52 and Pant-4 and at 50% FA in Sabha-5204. Accumulation of Fe, Si, Cu, Zn, Mn, Ni, Cd and As was investigated in roots, leaves and seeds of the plants. Fe accumulation was maximum in all the parts of plant followed by Si and both showed more translocation to leaves while Mn, Zn, Cu, Ni and Cd showed lower accumulation and most of the metal was confined to roots in all the three cultivars. As was accumulated only in leaves and was not found to be in detectable levels in roots and seeds. The metal accumulation order in three rice cultivars was Fe > Si > Mn > Zn > Ni > Cu > Cd > As in all the plant parts. The results showed that rice varieties Saryu-52 and Sabha-5204 were more tolerant and could show improved growth and yield in lower FA application doses as compared to Pant-4. Thus, Sabha-5204 and Saryu-52 are found suitable for cultivation in FA amended agricultural soils for better crop yields.     

Wood pellet fly ash and bottom ash as an effective liming agent and nutrient source for rye grass (Lolium perenne L.) and oats (Avena sativa)

Fly ash (FA) and bottom ash (BA) from a softwood pellet boiler were characterized and evaluated as soil amendments. In a greenhouse study, two plant species (rye grass, Lolium perenne L. and oats, Avena sativa) were grown in three different treatments (1% FA, 1% BA, non-amended control) of a silty loam soil. Total concentrations of plant nutrients Ca, K, Mg, P and Zn in both ashes were elevated compared to conventional wood ash. Concentrations of Cd, Cr, Pb, Se and Zn were found to be elevated in the FA relative to BA and the non-amended soil. At 28 d, oat above-ground biomass was found to be significantly greater in soil amended with FA. Potassium and Mo plant tissue concentrations were significantly increased by addition of either ash, and FA significantly increased Zn tissue concentrations. Cadmium and Hg tissue concentrations were elevated in some cases. As soil amendments, either pellet ash is an effective liming agent and nutrient source, but high concentrations of Cd and Zn in FA may preclude its use as an agricultural soil amendment in some jurisdictions. Lower ash application rates than those used in this study (i.e. <1%) may still provide sufficient nutrients and effective neutralization of soil acidity.     

A comparative study of cadmium phytoextraction by accumulator and weed species

Phytoextraction has shown great potential as an alternative technique for the remediation of metal contaminated soils. The objective of this study was to investigate cadmium (Cd) phytoextraction ability of high biomass producing weeds in comparison to indicator plant species. The pot study conducted with 10 to 200 mg Cd kg(-1) soil indicated that Ipomoea carnea was more effective in removing Cd from soil than Brassica juncea. Among the five species, B. juncea accumulated maximum Cd, but I. carnea followed by Dhatura innoxia and Phragmytes karka were the most suitable species for phytoextraction of cadmium from soil, if the whole plant or above ground biomass is harvested. In the relatively short time of this experiment, I. carnea produced more than 5 times more biomass in comparison to B. juncea. There were significant differences (p < 0.05) between the shoot length and shoot mass of control and treated plants.     

Effect of industrial waste products on phosphorus mobilisation and biomass production in abattoir wastewater irrigated soil

This study evaluated the effect of alkaline industrial by-products such as flyash (FA) and redmud (RM) on phosphorus (P) mobilisation in abattoir wastewater irrigated soils, using incubation, leaching and plant growth (Napier grass [Pennisetum purpureum]) experiments. The soil outside the wastewater irrigated area was also collected and treated with inorganic (KH₂PO₄ [PP]) and organic (poultry manure [PM]) P treatments, to study the effect of FA and RM on P mobilisation using plant growth experiment. Among the amendments, FA showed the highest increase in Olsen P, oxalic acid content and phosphatase activity. The highest increase in Olsen P for PM treated non-irrigated soils showed the ability of FA and RM in mobilising organic P better than inorganic P (PP). There was over 85 % increase in oxalic acid content in the plant growth soils compared to the incubated soil, showing the effect of Napier grass in the exudation of oxalic acid. Both amendments (FA and RM) showed an increase in phosphatase activity at over 90 % at the end of the 5-week incubation period. The leaching experiment indicated a decrease in water soluble P thereby ensuring the role of FA and RM in minimising P loss to water bodies. FA and RM showed an increase in plant biomass for all treatments, where FA amended soil showed the highest increase as evident from FA’s effect on Olsen P. Therefore, the use of FA and RM mobilised P in abattoir wastewater irrigated soils and increased biomass production of Napier grass plants through root exudation of oxalic acid.     

Effects of inoculation of plant growth-promoting rhizobacteria on metal uptake by Brassica juncea

A greenhouse study was carried out with Brassica juncea to critically evaluate effects of bacterial inoculation on the uptake of heavy metals from Pb-Zn mine tailings by plants. Application of plant growth-promoting rhizobacteria, including nitrogen-fixing bacteria and phosphate and potassium solubilizers, might play an important role in the further development of phytoremediation techniques. The presence of these beneficial bacteria stimulated plant growth and protected the plant from metal toxicity. Inoculation with rhizobacteria had little influence on the metal concentrations in plant tissues, but produced a much larger above-ground biomass and altered metal bioavailability in the soil. As a consequence, higher efficiency of phytoextraction was obtained compared with control treatments.     

Effect of metal tolerant plant growth promoting Bradyrhizobium sp. (vigna) on growth, symbiosis, seed yield and metal uptake by greengram plants

The nickel and zinc tolerant plant growth promoting Bradyrhizobium sp. (vigna) RM8 was isolated from nodules of greengram, grown in metal contaminated Indian soils. The plant growth promoting (PGP) potentials of strain RM8 was assessed both in the presence and absence of nickel and zinc under in vitro conditions. Strain RM8 tolerated a high level of nickel (300 microg ml(-1)) and zinc (1400 microg ml(-1)) on yeast extract mannitol agar medium. Bradyrhizobium sp. (vigna) strain RM8 produced 13.3 microg ml(-1) of indole acetic acid in Luria Bertani broth at 100 microg ml(-1) of tryptophan which increased to 13.6 microg ml(-1) at 50 microg Ni ml(-1) and 13.5 microg ml(-1) at 300 microg Zn ml(-1). Strain RM8 was positive for siderophore, HCN and ammonia both in the absence and presence of nickel and zinc. The PGP activity of this strain was further evaluated with increasing concentrations of nickel and zinc using greengram as a test crop. The bioinoculant enhanced the nodule numbers by 82%, leghaemoglobin by 120%, seed yield by 34%, grain protein by 13%, root N by 41% and shoot N by 37% at 290 mg Ni kg(-1) soil. At 4890 mg Zn kg(-1) soil, the bioinoculant increased the nodule numbers by 50%, leghaemoglobin by 100%, seed yield by 36%, grain protein by 13%, root N by 47% and shoot N by 42%. The bioinoculant strain RM8 reduced the uptake of nickel and zinc by plant organs compared to plants grown in the absence of bioinoculant. This study suggested that the bioinoculant due to its intrinsic abilities of growth promotion and attenuation of the toxic effects of nickel and zinc could be exploited for remediation of metal from nickel and zinc contaminated sites.     

Influence of sodium nitrilotriacetate (NTA) and citric acid on phenolic and organic acids in Brassica juncea grown in excess of cadmium

Brassica juncea cv. 426308 was grown in soils containing 150 mg Cd(2+)kg(-1) soil. After 38 days, the soil was amended with two rates of citric acid or NTA (10 and 20 mmol kg(-1) soil). Control soil was not amended with chelates. Plants were harvested during growth, immediately before and seven days after chelate addition. Shoot composition of organic and phenolic acids and shoot Cd(2+) concentration were determined. Cadmium concentration remained constant during the growth and increased following NTA and citric acid amendments depending on chelate type and concentration. The highest increments in Cd(2+) were measured after the addition of NTA. Compared to the control, 10 and 20 NTA-treated plants showed two- and three-fold increases, respectively. At 150 mg Cd(2+)kg(-1) soil the amount of organic and phenolic acids in the leaves of B. juncea was always higher than that detected in the control. A direct correlation between organic acid concentration and cadmium content was detected both during growth and after chelate addition. On the contrary, the amount of phenols seemed to be correlated with the metal content only in non-amended and NTA-treated plants. The 10 and 20 citric acid additions caused 45% and 90% increases in shoot phenolic acids although cadmium content rose to a smaller extent. The inhibition of citrate synthase and the entrance of phosphoenolpyruvate in shikimate pathway leading to the formation of aromatic compounds might come into play. The increase in phenylalanine ammonialyase activity following citric acid amendments suggested this metabolic response.     

Isolation and characterization of a heavy metal-resistant Burkholderia sp. from heavy metal-contaminated paddy field soil and its potential in promoting plant growth and heavy metal accumulation in metal-polluted soil

A heavy metal-resistant bacterial strain was isolated from heavy metal-contaminated soils and identified as Burkholderia sp. J62 based on the 16S rDNA gene sequence analysis. The heavy metal- and antibiotic resistance, heavy metal solubilization of the isolate were investigated. The isolate was also evaluated for promoting plant growth and Pb and Cd uptakes of the plants from heavy metal-contaminated soils in pot experiments. The isolate was found to exhibit different multiple heavy metal and antibiotic resistance characteristics. Atomic absorption spectrometer analysis showed increased bacterial solubilization of lead and cadmium in solution culture and in soils. The isolate produced indole acetic acid, siderophore and 1-aminocyclopropane-1-carboxylate deaminase. The isolate also solubilized inorganic phosphate. Inoculation with the isolate was found to significantly (p < 0.05) increase the biomass of maize and tomato plants. Increase in tissue Pb and Cd contents varied from 38% to 192% and from 5% to 191% in inoculated plants growing in heavy metal-contaminated soils compared to the uninoculated control, respectively. These results show that heavy metal-solubilizing and plant growth promoting bacteria are important for plant growth and heavy metal uptake which may provide a new microbial enhanced-phytoremediation of metal-polluted soils.     

Changes in amino acid profile and metal content in seeds of Cicer arietinum L. (chickpea) grown under various fly-ash amendments

Seeds of Cicer arietinum L. plants are edible and a valuable source of protein. Accumulation of toxic metals in the edible part of the plant, grown in fields close to fly-ash (FA) landfills, may pose a threat to human health. In the present study, the effects of FA and its amendments with different ameliorants viz., garden soil (GS), press mud (PM) and saw dust (SD), on total soluble protein contents, amino acid composition and metal accumulation in seeds were investigated in var. CSG-8962 and var. C-235 of C. arietinum. Plants accumulated adequate amounts of essential metals viz. Fe, Cu, Zn in seeds, while the toxic metals such as Cd and Cr were taken up in smaller quantities. The accumulation of Cr and Cd was less in var. C-235 than var. CSG-8962. Amendment of FA with PM enhanced the amount of soluble protein and amino acids in both varieties and was found to be superior among all tested ameliorants. Both quantitative and qualitative analysis of amino acids showed better response in var. C-235 as compared to var. CSG-8962. Thus var. C-235 seems to be suitable for cultivation in FA contaminated areas due to more accumulation of essential metals and less accumulation of toxic metals in seeds. Application of PM may further improve the growth of plants and nutritional quality of seeds.     

Nitrilotriacetate- and citric acid-assisted phytoextraction of cadmium by Indian mustard (Brassica juncea (L.) Czernj, Brassicaceae)

In a pot experiment the effects of nitrilotriacetate (NTA) and citric acid applications on Cd extractibility from soil as well as on its uptake and accumulation by Indian mustard (Brassica juncea) were investigated. Plants were grown in a sandy soil with added CdS at four levels ranging from 50 to 200 mg Cd kg(-1) soil. After 30 days of growth, pots were amended with NTA or citric acid at 10 and 20 mmol kg(-1). Control pots were not treated with chelates. Harvest of plants was performed immediately before and one week after chelate addition. Soil water-, NH(4)NO(3)- and EDTA-extractable Cd fractions increased constantly with both increasing soil metal application and chelate concentration. Shoot dry weights did not suffer significant reductions with increasing Cd addition to the soil except for both NTA treatments in which at 200 mg Cd kg(-1) a 30% decrease in dry matter was observed. Generally, following NTA and citric acid amendments, Cd concentration in shoots increased with soil Cd level. However, due to Cd toxicity, at the highest metal application rate both NTA treatments lowered Cd concentration in the above-ground parts. Compared to the control, at 10 mmol kg(-1) citric acid did not change Cd concentration in shoots, whereas NTA-treated plants showed an about 2-fold increase. The addition of chelates at 20 mmol kg(-1) further enhanced Cd concentration in shoots up to 718 and 560 microg g(-1) dry weight in the NTA and citrate treatments, respectively.     

Translocation of metals from fly ash amended soil in the plant of Sesbania cannabina L. Ritz: effect on antioxidants

The plants of Sesbania cannabina Ritz grown on different amendments of fly ash (FA), have shown a high accumulation of metals (Fe, Mn, Zn, Cu, Pb and Ni). The highest accumulation of Fe the and lowest level of Ni were recorded in these plants. The different amendments of fly ash with garden soil (GS) were extracted with DTPA and the levels of metals were found to be decreased with an increase in fly ash application ratio from 10% to 50% FA. The analysis of the results showed an increase in the level of malondialdehyde (MDA) content of the roots for all the exposure periods. The maximum increases of 136% (roots) and 120% (leaves) were observed in MDA content at 100% FA after 30 d of growth of the plant, compared to GS. The level of antioxidants was found to increase for all the exposure periods in the roots of the plants to combat metal stress. At 30 d, the maximum increase of 57% (ascorbic acid) and 78% (free proline) was observed in the roots of the plants grown on 100% and 10% FA, respectively, as compared to their respective GS. At 90 d, a maximum increase of 42% (cysteine) and 117% (NPSH) was recorded in the roots of the plants grown on 25% and 100% FA, respectively, as compared to their respective GS. In leaves, a significant increase in antioxidants i.e. cysteine, NPSH and free proline content was recorded after 30 d, whereas no such trend was observed for the rest of the exposure periods. The chlorophyll and carotenoid contents increased with an increase in the FA amendment ratio from 10% to 50% FA for all the exposure periods as compared to GS. In both roots and leaves, the level of protein content increased in all the amendments and 100% FA at 30 d as compared to GS. Thus, there is a balance in the level of MDA content and level of antioxidants in the plants at 90 d. In view of its tolerance, the plants may be used for phytoremediation of metals from fly ash contaminated sites and suitable species for plantation on fly ash land fills.     

Influence of metal resistant-plant growth-promoting bacteria on the growth of Ricinus communis in soil contaminated with heavy metals

The metal resistant-plant growth-promoting bacterial (PGPB) strains PsM6 and PjM15 isolated from a serpentine soil were characterized as Pseudomonas sp. and Pseudomonas jessenii, respectively, on the basis of their morphological, physiological, biochemical characteristics and 16S rDNA sequences. Assessment of plant growth-promoting parameters revealed the intrinsic ability of the strains for the utilization of 1-aminocyclopropane-1-carboxylic acid as the sole N source, solubilization of insoluble phosphate and production of indole-3-acetic acid (IAA). Further, a pot experiment was conducted to elucidate the effects of inoculating metal resistant PGPB on the plant growth and the uptake of Ni, Cu and Zn by Ricinus communis. Inoculation of Pseudomonas sp. PsM6 or P. jessenii PjM15 increased the shoot and root biomass of R. communis grown in non-contaminated and contaminated soil. However, the maximum biomass was observed in the plants inoculated with strain PjM15. This effect can be attributed to the solubilization of phosphate and production of IAA. Inoculation of Pseudomonas sp. PsM6 and PjM15 did not greatly alter the organ metal concentrations except Zn which concentration was higher in root, stem and leaf of inoculated plants. The results of metal extraction with PGPB strains showed that PsM6 was more efficient at solubilizing Zn than PjM15, and that PjM15 was better at solubilising Ni and Cu than PsM6. Owing to its wide action spectrum, the metal resistant PGPB could serve as an effective metal sequestering and growth-promoting bioinoculant for plants in metal-stressed soil. The present study has provided a new insight into the phytoremediation of metal-contaminated soil.     

A remediation strategy based on active phytoremediation followed by natural attenuation in a soil contaminated by pyrite waste

Phytoremediation of metal-polluted soils can be promoted by the proper use of soil amendments and agricultural practices. A 4-year phytoremediation programme was applied to a site affected by the toxic spill of pyrite residue at Aznalcóllar (Spain) in 1998, contaminated with heavy metals (Zn, Cu, Pb, Cd) and arsenic. This consisted of active phytoremediation, using organic amendments (cow manure and compost) and lime and growing two successive crops of Brassica juncea (L.) Czern., followed by natural attenuation without further intervention. Changes in soil pH, extractable metal and As concentrations, organic carbon content and microbial biomass was evaluated. The initial oxidation of metal sulphides from pyrite residues released soluble metals and reduced soil pH to extremely acidic values (mean 4.1, range 2.0-7.0). The addition of lime (up to 64 t ha(-1)) increased soil pH to adequate values for plant growth, resulting in a significant decrease in DTPA-extractable metal concentrations in all plots. The natural attenuation phase showed also a decrease in extractable metals. Organic treatments increased the soil total organic carbon, which led to higher values of microbial biomass (11.6, 15.2 and 14.9 g kg(-1) TOC and 123, 170 and 275 microg g(-1) biomass-C in control, compost and manure plots, respectively). Active phytoremediation followed by natural attenuation, was effective for remediation of this pyrite-polluted soil.