Enhanced phytoextraction: I. Effect of EDTA and citric acid on heavy metal mobility in a calcareous soil

Phytoextraction, the use of plants to extract heavy metals from contaminated soils, could be an interesting alternative to conventional remediation technologies. However, calcareous soils with relatively high total metal contents are difficult to phytoremediate due to low soluble metal concentrations. Soil amendments such as ethylene diaminetetraacetate (EDTA) have been suggested to increase heavy metal bioavailability and uptake in aboveground plant parts. Strong persistence of EDTA and risks of leaching of potentially toxic metals and essential nutrients have led to research on easily biodegradable soil amendments such as citric acid. In our research, EDTA is regarded as a scientific benchmark with which degradable alternatives are compared for enhanced phytoextraction purposes. The effects of increasing doses of EDTA (0.1,1,10 mmol kg(-1) dry soil) and citric acid (0.01, 0.05, 0.25, 0.442, 0.5 mol kg(-1) dry soil) on bioavailable fractions of Cu, Zn, Cd, and Pb were assessed in one part of our study and results are presented in this article. The evolution of labile soil fractions of heavy metals over time was evaluated using water paste saturation extraction (approximately soluble fraction), extraction with 1 M NH4OAc at pH 7 (approximately exchangeable fraction), and extraction with 0.5 M NH4OAc + 05 M HOAc + 0.02 M EDTA at pH 4.65 (approximately potentially bioavailable fraction). Both citric acid and EDTA produced a rapid initial increase in labile heavy metal fractions. Metal mobilization remained constant in time for soils treated with EDTA, but a strong exponential decrease of labile metal fractions was noted for soils treated with citric acid. The half life of heavy metal mobilization by citric acid varied between 1.5 and 5.7 d. In the following article, the effect of heavy metal mobilization on uptake by Helianthus annuus will be presented.     

Phytoextraction of copper from contaminated soil by Elsholtzia splendens as affected by EDTA, citric acid, and compost

Phytoextraction of copper (Cu) from contaminated soils greatly depends on the metal bioavailability in the soils and metal uptake ability of the plant. In this study, the effects of chelators [ethylenediamine tetraacetic acid (EDTA), citric acid (CA)] and compost amendments on Cu phytoextraction potential by a tolerant and accumulating plant species (E. splendens) were examined in two types of contaminated soils, ie., the mined soil from Cu-mined area (MS) and a paddy soil polluted by Cu refining (PS). The results showed that EDTA application at 2.5-5.0 mmol kg(-1) increased phytoextraction of Cu by four- and eight-fold from both MS and PS, respectively, which is mainly attributed to increased H2O extractable Cu in the soil. The Cu amount extracted by the shoots of E. splendens reached 800-1000 microg Cu plant(-1) from the MS and 400-700 microg Cu plant(-1) from the PS at EDTA application rates of 2.5-5.0 mmol kg(-1). The application of CA at 5.0 mmol kg(-1) had minimal effects on Cu extractability in both soils and slightly decreased Cu extraction efficiency by E. splendens. Plant biomass production was enhanced by CA at 0.25 mmol L(-1) in nutrient solution, but inhibited by CA at 5.0 mmol kg(-1) in both MS and PS. Increasing the compost rate significantly decreased H2O extractable Cu in the MS, but raised H2O-extractable Cu in the PS, which resulted mainly front the reduced exchangeable Cu in the MS and the increased exchangeable and organic fractions of Cu in the PS by compost. At high compost rate (5%), the shoots of E. splendens extracted 3.6-fold higher Cu from the PS than from the MS. These results indicate that, among the soil amendments, efficiency of Cu phytoextraction is enhanced mostly by 2.5-5.0 mmol kg(-1) EDTA, followed by 5% (w:w) compost, whereas < 5.0 mmol kg(-1) CA has minimal effects on Cu phytoextraction by E. splendens in the PS. As for the MS, only 2.5-5.0 mmol kg(-1) EDTA can elevate the efficiency of Cu, while 5% compost amendment and < 5.0 mmol kg(-1) CA application have no marked effects on Cu phytoextraction by E. splendens.     

EDTA enhanced heavy metal phytoextraction: metal accumulation,leaching and toxicity

Synthetic chelates such as ethylene diamine tetraacetic acid (EDTA) have been shown to enhance phytoextraction of some heavy metals from contaminated soil. In a soil column study, we examined the effect of EDTA on the uptake of Pb, Zn and Cd by Chinese cabbage (Brassica rapa), mobilization and leaching of heavy metals and the toxicity effects of EDTA additions on plants. The most effective was a single dose of 10 mmol EDTA kg^–1 soil where we detected Pb, Zn and Cd concentrations that were 104.6, 3.2 and 2.3-times higher in the aboveground plant biomass compared to the control treatments. The same EDTA addition decreased the concentration of Pb, Zn and Cd in roots of tested plants by 41, 71 and 69%, respectively compared to concentrations in the roots of control plants. In columns treated with 10 mmol kg^–1 EDTA, up to 37.9, 10.4 and 56.3% of initial total Pb, Zn and Cd in soil were leached down the soil profile, suggesting high solubility of heavy metals-EDTA complexes. EDTA treatment had a strong phytotoxic effect on the red clover (Trifolium pratense) in bioassay experiment. Moreover, the high dose EDTA additions inhibited the development of arbuscular mycorrhiza. The results of phospholipid fatty acid analyses indicated toxic effects of EDTA on soil fungi and increased environmental stress of soil microfauna.     

Enhanced phytoextraction: in search of EDTA alternatives

Enhanced phytoextraction proposes the use of soil amendments to increase the heavy-metal content of above-ground harvestable plant tissues. This study compares the effect of synthetic aminopolycarboxylic acids [ethylenediamine tetraacetatic acid (EDTA), nitriloacetic acid (NTA), and diethylenetriamine pentaacetic acid (DTPA)] with a number of biodegradable, low-molecular weight, organic acids (citric acid, ascorbic acid, oxalic acid, salicylic acid, and NH4 acetate) as potential soil amendments for enhancing phytoextraction of heavy metals (Cu, Zn, Cd, Pb, and Ni) by Zea mays. The treatments in this study were applied at a dose of 2 mmol/kg(-1) 1 d before sowing. To compare possible effects between presow and postgermination treatments, a second smaller experiment was conducted in which EDTA, citric acid, and NH4 acetate were added 10 d after germination as opposed to 1 d before sowing. The soil used in this screening was a moderately contaminated topsoil derived from a dredged sediment disposal site. This site has been in an oxidized state for more than 8 years before being used in this research. The high carbonate, high organic matter, and high clay content characteristic to this type of sediment are thought to suppress heavy-metal phytoavailability. Both EDTA and DTPA resulted in increased levels of heavy metals in the above-ground biomass. However, the observed increases in uptake were not as large as reported in the literature. Neither the NTA nor organic acid treatments had any significant effect on uptake when applied prior to sowing. This was attributed to the rapid mineralization of these substances and the relatively low doses applied. The generally low extraction observed in this experiment restricts the use of phytoextraction as an effective remediation alternative under the current conditions, with regard to amendments used, applied dose (2 mmol/kg(-1) soil), application time (presow), plant species (Zea mays), and sediment (calcareous clayey soil) under study.     

Effect of drought, on transport and metabolism of abscisic acid in aeroponically grown Helianthus annuus

The effect of drought on transport and metabolism of radioactive abscisic acid (ABA) in roots and shoots of sunflower ( Helianthus annuus L. cv. Russian) was observed. Radioactivity from ABA moved freely all over the plants. Young shoot tissues, such as the growing apical bud or axillary buds released from apical dominance, were strong sinks for ABA. Mature tissues were effective exporters. Drought-induced alterations in the pattern of transport of radioactivity do not appear to be a major factor in the control of drought-induced changes in ABA levels. Metabolism of ABA occurred in all organs examined in stressed and unstressed plants. Labelled ABA and its metabolites moved in the xylem. Drought altered the quantity of radioactive metabolites and reduced the amount of radioactive ABA in extracts from the stressed plants.     

Effect of pre-treatment with ethanolamine on the response of Helianthus annuus L. to salt stress

The accumulation of compatible solutes is one of the strategies that plants have developed to tolerate salt stress. Proline and betaine are the main metabolites that accumulate in various species of higher plants in response to salt stress. In Helianthus annuus L., pre-treatment of seeds with ethanolamine led to enhanced seedling tolerance to conditions of saline stress during germination, as evidenced by the greater growth of pretreated seedlings (EAS group) versus untreated seedlings (S group), evaluated through such parameters as length, water and chlorophyll content. During the germination period, a considerable increase was observed in proline levels (up to 300%) in seedlings subjected to saline stress, whereas in the EAS group, the proline increment was much smaller (20%). Starting from the fourth day of germination, betaine levels in seedlings pretreated with ethanolamine and then with water (EAW group) and in EAS showed a significant increase versus C (control) and S seedlings, possibly because such a precursor promotes betaine biosynthesis. This could be responsible for the enhanced growth observed in EAS versus S seedlings, as well as for preventing the decrease in chlorophyll content in the EAS group. The accumulation of betaine seems to correlate with the greater tolerance of these seedlings against stress induced by sodium chloride.     

Impact of chelator-induced phytoextraction of cadmium on yield and ionic uptake of maize

Enhanced phytoextraction uses soil chelators to increase the bioavailability of heavy metals. This study tested the effectiveness of ethylenediaminetetraacetic acid (EDTA) and citric acid in enhancing cadmium (Cd) phytoextraction and their effects on the growth, yield, and ionic uptake of maize (Zea mays). Maize seeds of two cultivars were sown in pots treated with 15 (Cd₁₅) or 30 mg Cd kg^?1 soil (Cd₃₀). EDTA and citric acid at 0.5 g kg^?1 each were applied 2 weeks after germination. Results demonstrated that the growth, yield per plant, and total grain weight were reduced by exposure to Cd. EDTA increased the uptake of Cd in shoots, roots, and grains of both maize varieties. Citric acid did not enhance the uptake of Cd, rather it ameliorated the toxicity of Cd, as shown by increased shoot and root length and biomass. Cadmium toxicity reduced the number of grains, rather than the grain size. The maize cultivar Sahiwal-2002 extracted 1.6% and 3.6% of Cd from soil in both Cd+ EDTA treatments. Hence, our study implies that maize can be used to successfully phytoremediate Cd from soil using EDTA, without reducing plant biomass or yield.     

Stomatal response to drying soil in relation to changes in the xylem sap composition of Helianthus annuus. II. Stomatal sensitivity to abscisic acid imported from the xylem sap

Sunflower plants [Helianthus annuus L.) were subjected to soil drought. Leaf conductance declined with soil water content even when the shoot was kept turgid throughout the drying period. The concentration of abscisic acid in the xylem sap increased with decreasing soil water content. No general relation could be established between abscisic acid concentration in the xylem sap and leaf conductance due to marked differences in the sensitivity of leaf conductance of individual plants to abscisic acid from the xylem sap. The combination of these results with data from Gollan, Schurr & Schulze (1992, see pp. 551–559, this issue) reveals close connection of the effectiveness of abscisic acid as a root to shoot signal to the nutritional status of the plant.     

海水胁迫对向日葵苗期生长及矿质营养吸收特性的影响

采用砂培法,研究了海水胁迫对向日葵幼苗生长及矿质营养吸收特性的影响。结果表明,海水胁迫下,向日葵幼苗株高、茎粗、干物重明显降低。幼苗根茎叶中Cl-,茎和叶中Mg2 、叶中Na 和Ca2 含量随海水浓度的增加而增加,根茎叶中K 、全氮和全磷含量随海水浓度升高而降低,但在10%和20%海水胁迫下,向日葵体内Na 、Cl-主要集中于根和茎中,叶中较少。海水胁迫下,向日葵幼苗各部位K /Na 始终是叶部最高,根部最低,且根茎叶中SK,Na值均大于1。因此,低浓度海水胁迫下向日葵幼苗对Na 和Cl-的截流作用、海水胁迫下幼苗根部对K 强的选择性吸收以及K 向地上部的选择性运输是向日葵具有一定耐盐性的主要原因。     

Enhanced Pb Absorption by Hordeum vulgare L. and Helianthus annuus L. Plants Inoculated with an Arbuscular Mycorrhizal Fungi Consortium

The effect of an arbuscular mycorrhizal fungi (AMF) consortium conformed by (Glomus intraradices, Glomus albidum, Glomus diaphanum, and Glomus claroideum) on plant growth and absorption of Pb, Fe, Na, Ca, and ³²P in barley (Hordeum vulgare L.) and sunflower (Helianthus annuus L.) plants was evaluated. AMF-plants and controls were grown in a substrate amended with powdered Pb slag at proportions of 0, 10, 20, and 30% v/v equivalent to total Pb contents of 117; 5,337; 13,659, and 19,913 mg Pb kg^?1 substrate, respectively. Mycorrhizal root colonization values were 70, 94, 98, and 90%, for barley and 91, 97, 95, and 97%, for sunflower. AMF inoculum had positive repercussions on plant development of both crops. Mycorrhizal barley absorbed more Pb (40.4 mg Pb kg^?1) shoot dry weight than non-colonized controls (26.5 mg Pb kg^?1) when treated with a high Pb slag dosage. This increase was higher in roots than shoots (650.0 and 511.5 mg Pb kg^?1 root dry weight, respectively). A similar pattern was found in sunflower. Plants with AMF absorbed equal or lower amounts of Fe, Na and Ca than controls. H. vulgare absorbed more total P (1.0%) than H. annuus (0.9%). The arbuscular mycorrizal consortium enhanced Pb extraction by plants.     

Root development and heavy metal phytoextraction efficiency: comparison of different plant species in the field

Heavy metal phytoextraction is a soil remediation technique which implies the optimal use of plants to remove contamination from soil. Plants must thus be tolerant to heavy metals, adapted to soil and climate characteristics and able to take up large amounts of heavy metals. Their roots must also fit the spatial distribution of pollution. Their different root systems allow plants to adapt to their environment and be more or less efficient in element uptake. To assess the impact of the root system on phytoextraction efficiency in the field, we have studied the uptake and root systems (root length and root size) of various high biomass plants (Brassica juncea, Nicotiana tabacum, Zea mays and Salix viminalis) and one hyperaccumulator (Thlaspi caerulescens) grown in a Zn, Cu and Cd contaminated soil and compared them with total heavy metal distribution in the soil. Changes from year to year have been studied for an annual (Zea mays) and a perennial plant (Salix viminalis) to assess the impact of the climate on root systems and the evolution of efficiency with time and growth. In spite of a small biomass, T. caerulescens was the most efficient plant for Cd and Zn removal because of very high concentrations in the shoots. The second most efficient were plants combining high metal concentrations and high biomass (willows for Cd and Zn and tobacco for Cu and Cd). A large cumulative root density/aboveground biomass ratio (L_A/B), together with a relative larger proportion of fine roots compared to other plants seemed to be additional favourable characteristics for increased heavy metal uptake by T. caerulescens. In general, for all plants correlations were found between L _A/B and heavy metal concentrations in shoots (r=0.758^***, r=0.594^***, r=0.798^*** (P<0.001) for Cd, Cu and Zn concentrations resp.). Differences between years were significant because of variations in climatic conditions for annual plants or because of growth for perennial plants. The plants exhibited also different root distributions along the soil profile: T. caerulescens had a shallow root system and was thus best suited for shallow contamination (0.2 m) whereas maize and willows were the most efficient in colonising the soil at depth and thus more applicable for deep contamination (0.7 m). In the field situation, no plant was able to fit the contamination properly due to heterogeneity in soil contamination. This points out to the importance and the difficulty of choosing plant species according to depth and heterogeneity of localisation of the pollution.     

Effects of citric acid and the siderophore desferrioxamine B (DFO-B) on the mobility of germanium and rare earth elements in soil and uptake in Phalaris arundinacea

Effects of citric acid and desferrioxamine B (DFO-B) on the availability of Ge and selected rare earth elements (REEs) (La, Nd, Gd, Er) to Phalaris arundinacea were investigated. A soil dissolution experiment was conducted to elucidate the effect of citric acid and DFO-B at different concentrations (1 and 10 mmol L^?1 citric acid) on the release of Ge and REEs from soil. In a greenhouse, plants of P. arundinacea were cultivated on soil and on sand cultures to investigate the effects of citric acid and DFO-B on the uptake of Ge and REEs by the plants. Addition of 10 mmol L^?1 citric acid significantly enhanced desorption of Ge and REEs from soil and uptake into soil-grown plants. Applying DFO-B enhanced the dissolution and the uptake of REEs, while no effect on Ge was observed. In sand cultures, the presence of citric acid and DFO-B significantly decreased the uptake of Ge and REEs, indicating a discrimination of the formed complexes during uptake. This study clearly indicates that citric acid and the microbial siderophore DFO-B may enhance phytoextraction of Ge and REEs due to the formation of soluble complexes that increase the migration of elements in the rhizosphere.     

Cloning,sequence and characterization of a sunflower (Helianthus annuus L.) pathogen-induced gene showing sequence homology with auxin-induced genes from plants

The establishment of a plant-pathogen interaction involves changes in gene expressions in both organisms. To isolate Helianthus annuus genes whose expression is induced during processes of resistance to Plasmopara halstedii, a comparison of the expression pattern of healthy sunflowers was made with sunflowers infected with 2 races of P. halstedii, either virulent or avirulent, using differential display of mRNA. A full-length cDNA, HaAC1, representing a sunflower gene whose expression is enhanced during early stages of the incompatible interaction, was isolated. Different timing of RNA accumulation is observed between compatible and incompatible combinations. Sequence analysis and database search revealed significant homology with auxin-induced genes from plants. The expression of this gene, is also induced after treatment with 2,4-dichlorophenoxyacetic acid (2,4-D), salicylic acid (SA) and wounding.     

Immunofluorescent Localization of a Connexin 26-like Protein at the Surface of Mesophyll Protoplasts from Vicia faba L. and Helianthus annuus L.

Mesophyll protoplasts from three week old leaves of Helianthus annuus L. and from four week old leaves of Vicia faba L. were incubated with polyclonal, monospecific antibodies, raised against either cx 32 or cx 26 mouse liver connexin. Crossreactions were visualized by FITC-labeled anti-rabbit antibodies. Incubations with the cx 26 antibody resulted in fluorescing spots on protoplast surfaces of both plant species, indicating the presence of a polypeptide, immunologically related to the animal cx 26. A plant protein, exhibiting similarities to cx 26, would present a new member of connexin-like plant proteins. Controls, performed with preimmune serum or with the FITC-conjugate alone, were negative. Immunofluorescing spots were not obtained after incubations with the cx 32 antibody. Since the existence of a cx 32-like plant protein, associated with ultrastructures of plasmodesmata and the plasma membrane, is meanwhile established, several explanations for the failed attempt to demonstrate a cx 32 antibody labeling at protoplast surfaces are discussed.     

Comparison of EDTA- and Citric Acid-Enhanced Phytoextraction of Heavy Metals in Artificially Metal Contaminated Soil by Typha Angustifolia

A pot experiment was conducted to study the performance of EDTA and citric acid (CA) addition in improving phytoextraction of Cd, Cu, Pb, and Cr from artificially contaminated soil by T. angustifolia. T. angustifolia showed the remarkable resistance to heavy metal toxicity with no visual toxic symptom including chlorosis and necrosis when exposed to metal stress. EDTA-addition significantly reduced plant height and biomass, compared with the control, and stunted plant growth, while 2.5 and 5 mM CA addition induced significant increases in root dry weight. EDTA, and 5 and 10 mM CA significantly increased shoot Cd, Pb, and Cr concentrations compared with the control, with EDTA being more effective. At final harvest, the highest shoot Cd, Cr, and Pb concentrations were recorded in the treatment of 5 mM EDTA addition, while maximal root Pb concentration was found at the 2.5 mM CA treatment. However, shoot Cd accumulation in the 10 mM CA treatment was 36.9% higher than that in 2.5 mM EDTA, and similar with that in 10 mM EDTA. Shoot Pb accumulation was lower in 10 mM CA than that in EDTA treatments. Further, root Cd, Cu, and Pb accumulation of CA treatments and shoot Cr accumulation in 5 or 10 mM CA treatments were markedly higher than that of control and EDTA treatments. The results also showed that EDTA dramatically increased the dissolution of Cu, Cr, Pb, and Cd in soil, while CA addition had less effect on water-soluble Cu, Cr, and Cd, and no effect on Pb levels. It is suggested that CA can be a good chelator candidate for T. angustifolia used for environmentally safe phytoextraction of Cd and Cr in soils.     

Chemically enhanced phytoextraction of risk elements from a contaminated agricultural soil using Zea mays and Triticum aestivum: performance and metal mobilization over a three year period

Enhanced phytoextraction using EDTA for the remediation of an agricultural soil contaminated with less mobile risk elements Cd and Pb originating from smelting activities in Príbram (Czech Republic) was assessed on the laboratory and the field scale. EDTA was applied to the first years crop Zea mays. Metal mobilization and metal uptake by the plants in the soil were monitored for two additional years when Triticum aestivum was planted. The application ofEDTA effectively increased water-soluble Cd and Pb concentrations in the soil. These concentrations decreased over time. Anyhow, increased concentrations could be still observed in the third experimental year indicating a low possibility of groundwater pollution after the addition of EDTA during and also after the enhanced phytoextraction process under prevailing climatic conditions. EDTA-applications caused phytotoxicity and thereby decreased biomass production and increased Cd and Pb uptake by the plants. Phytoextraction efficiency and phytoextraction potential were too low for Cd and Pb phytoextraction in the field in a reasonable time frame (as less than one-tenth of a percent of total Cd and Pb could be removed). This strongly indicates that EDTA-enhanced phytoextraction as implemented in this study is not a suitable remediation technique for risk metal contaminated soils.     

Removal of Heavy Metals from Soil Polluted with Effluents from a Paint Industry Using Helianthus annuus L. and Tithonia diversifolia (Hemsl.) as Influenced by Fertilizer Applications

Contamination of soils by effluents from industries is on the increase. There is the possibility of remediating these contaminated soils through the use of certain plants. This work investigated the remediating ability of Helianthus annuus and Tithonia diversifolia on the soil polluted with effluents from a paint industry in Ibadan, Nigeria. The experiment consisted of three treatments (H. annuus, T. diversifolia, and control) each replicated three times in a factorial combination of four different fertility managements, viz mineral fertilizer (MF); Grade A organomineral fertilizer (OMF); control₁, plants without fertilizer application; and control₂, where no fertilizer and no crop was planted using randomized complete block design. A total of 12 plots of 2 × 4 m² each per phytoplant were obtained. Each plot was planted with the viable seeds of the phytoplant at a spacing of 60 × 30 cm² and at the seed rate of four seeds per hole. The seedlings were thinned to two stands per hole 2 weeks after planting (WAP) and also weeded two times (2 and 5 WAP). After in situ second successive cultivation, percentage removal of heavy metals by Helianthus annuus with MF and OMF, respectively, were Cu 32.5 and 41.6; Pb 30.3 and 42.8; and Cd 44.5 and 56.7. Tithonia diversifolia, similarly, removed, respectively, Cu 16.9 and 23.4; Pb 36.9 and 43.7; and Cd 20.1 and 35.1. Lower percentages were removed in the controls where no fertilizer was applied. In the shoot of H. annuus with OMF, significantly (p< .05) higher values of 0.27, 1.72, and 0.11 mg kg^?1 of Cu, Pb, and Cd, respectively, were removed and stored at second cultivation as against 0.21, 3.39 and 0.08 mg kg^?1 in the shoot of T diversifolia. Lower values of Cu, Pb, and Cd were removed with MF, and also at first cultivation with OMF and MF. This study therefore recommends the use of sunflower plants, whether hybrids or wild-types along with the application of OMF for the effective remediation of soils contaminated with heavy metals, particularly in tropical climate.     

Heavy metals uptake by vegetables growing in sewage irrigated soil: relationship with heavy metal fractionation in soil

Abstract

Concentrations of the heavy metals Cd, Ni, Cr, Zn, Mn, Co and Cu in soils and vegetable samples (i.e. green chili and gourd) taken from six vegetable fields in the vicinity of Lahore, Pakistan were measured. These soils have been irrigated for a long time with untreated sewage effluents. A control site was selected that has history of fresh canal water irrigation. The sequential extraction procedure developed by Tessier was adopted to demarcate five metal fractions: exchangeable, acid soluble, reducible, oxidizable and residual. The extractants and digests were analysed by atomic absorption spectrometry. The fractionation procedure showed that all the metals were dominant (>50%) in the residual phase in control as well as in waste waster irrigated soils. The concentrations of all the metals in edible parts of the vegetables collected from wastewater irrigated soils, were above critical levels. The total metal concentrations in wastewater irrigated soils followed the order Mn> Co> Zn> Cr> Ni> Cu> Cd, while in control soils the ord er was Mn> Zn> Ni> Co> Cr> Cu> Cd. In order to understand the uptake of metals from soil to vegetables, correlation analyses were performed between metal concentrations in different fractions of soil and their concentrations in vegetables. Correlation analysis was performed at 95% and 99% confidence level. The meaningful significant negative correlation was observed between Cd_F5–Cd_chili,Cr_Total–Cr_gourd,Mn_F5–Mn_Chili,Gourd, Co_F5, _Total–Co_chili, Cu_Total–Cu_Chili which indicate the non availability of total content and residual fraction to studied vegetables. The positive correlation was observed between Cd_Fl-Cd_ChiU, Cd_Fl–Cd_gourd, Ni_{Fi, F3}–Ni_Gourd, Cr_{Fi, F3}, _F4–Cr_ChiU, Cr_F2–Cr_Gourd, Zn_{Fi, F2}–Zn_Gourd, Mn_F3–Mn_Chili, Mn_Fi–Mn_Gourd, Co_F2–Co_Chili, Cu_Fi–Cu_Chili. The positive correlation, especially for the first (exchangeable) second (acid soluble) and for the third (reducible) extraction steps, was obtained which indicate the bioavailability of these metal fractions to plant.     

Effect of an artificial metal pollution on nematode assemblage of a calcareous loamy chernozem soil

Heavy metal pollution in soils constitutes an acute problem threatening various groups of living organisms. Nematode samples were taken in the framework of a complex food-chain experiment which is being carried out after artificially contaminating a part of an agrobiotope. The 13 heavy metals and microelements involved in the study are the following: Al, As, Ba, Cd, Cr, Cu, Hg, Mo, Ni, Pb, Se, Sr, and Zn. Parallel plots receiving maximum concentrations equal to approx. 270 ppm were sampled twice during a complete vegetation period of winter wheat. Nematode abundance and richness as well as Maturity Index data revealed that Se and Cr had significantly negative effects on assemblages, while Zn appeared to have a generally positive influence on these parameters. Impacts by the other elements are less obvious or quite neutral (like that of Al). Favorable soil conditions and time elapsed (over 5 years) since the contamination, both resulting in a decrease of available element concentrations, may account for the not very pronounced effects. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effect of bacterial inoculation of strains of pseudomonas aeruginosa,alcaligenes feacalis and bacillus subtilis on germination,growth and heavy metal (cd,cr, and ni) uptake of brassica juncea

Bacterial inoculation may influence Brassica juncea growth and heavy metal (Ni, Cr, and Cd) accumulation. Three metal tolerant bacterial isolates (BCr3, BCd33, and BNi11) recovered from mine tailings, identified as Pseudomonas aeruginosa KP717554, Alcaligenes feacalis KP717561, and Bacillus subtilis KP717559 were used. The isolates exhibited multiple plant growth beneficial characteristics including the production of indole-3-acetic acid, hydrogen cyanide, ammonia, insoluble phosphate solubilization together with the potential to protect plants against fungal pathogens. Bacterial inoculation improved seeds germination of B. juncea plant in the presence of 0.1 mM Cr, Cd, and Ni, as compared to the control treatment. Compared with control treatment, soil inoculation with bacterial isolates significantly increased the amount of soluble heavy metals in soil by 51% (Cr), 50% (Cd), and 44% (Ni) respectively. Pot experiment of B. juncea grown in soil spiked with 100 mg kg^?1 of NiCl₂, 100 mg kg^?1 of CdCl₂, and 150 mg kg^?1 of K₂Cr₂O₇, revealed that inoculation with metal tolerant bacteria not only protected plants against the toxic effects of heavy metals, but also increased growth and metal accumulation of plants significantly. These findings suggest that such metal tolerant, plant growth promoting bacteria are valuable tools which could be used to develop bio-inoculants for enhancing the efficiency of phytoextraction.