Heavy metal tolerance genes: prospective tools for bioremediation

Unlike compounds that can be broken down, the remediation of most heavy metals and radionuclides requires removal from contaminated sources. Plants can extract inorganics, but effective phytoextraction requires plants that produce high biomass, grow rapidly and possess high capacity-uptake for the inorganic substrate. Either the existing hyperaccumulator plants must be bred for increased growth and biomass, or that hyperaccumulation traits must be engineered into fast growing, high biomass plants. The latter approach requires fundamental knowledge of the molecular mechanisms in the uptake and storage of inorganics. Much has been learned in recent years on how plants and certain fungi chelate and transport cadmium. This progress has been facilitated by the use of Schizosaccharomyces pombe as a model system. As target genes are identified in a model organism, their sequences can be modified for expression in a heterologous host or aid in the search of homologous genes in more complex organisms. Moreover, as plant nutrient uptake is intrinsically linked to the association with rhizospheric fungi, elucidating metal sequestration in this fungus permits additional opportunities for engineering rhizospheric microbes to assist in phytoextraction.     

The effectiveness of arbuscular-mycorrhizal fungi and Aspergillus niger or Phanerochaete chrysosporium treated organic amendments from olive residues upon plant growth in a semi-arid degraded soil

Arbuscular mycorrhizal (AM) fungi and a residue from dry olive cake (DOC) supplemented with rock phosphate (RP) and treated with either Aspergillus niger (DOC-A) or Phanerochaete chrysosporium (DOC-P), were assayed in a natural, semi-arid soil using Trifolium repens or Dorycnium pentaphyllum plants. The effects of the AM fungi and/or DOC-A were compared with P-fertilisation (P) over eleven successive harvests to evaluate the persistence of the effectiveness of the treatments. The biomass of dually-treated plants after four successive harvests was greater than that obtained for non-treated plants or those receiving the AM inoculum or DOC-A treatments after eleven yields. The AM inoculation was critical for obtaining plant growth benefit from the application of fermented DOC-A residue. The abilities of the treatments to prevent plant drought stress were also assayed. Drought-alleviating effects were evaluated in terms of plant growth, proline and total sugars concentration under alternative drought and re-watering conditions (8th and 9th harvests). The concentrations of both compounds in plant biomass increased under drought when DOC-A amendment and AM inoculation were employed together: they reinforced the plant drought-avoidance capabilities and anti-oxidative defence. Water stress was less compensated in P-fertilised than in DOC-A-treated plants. DOC-P increased D. pentaphyllum biomass, shoot P content, nodule number and AM colonisation, indicating the greater DOC-transforming ability of P. chrysosporium compared to A. niger. The lack of AM colonisation and nodulation in this soil was compensated by the application of DOC-P, particularly with AM inoculum. The management of natural resources (organic amendments and soil microorganisms) represents an important strategy that assured the growth, nutrition and plant establishment in arid, degraded soils, preventing the damage that arises from limited water and nutrient supply.     

Biohythane production from pineapple peel using Methanosarcina mazei enhanced with palm oil mill effluent (POME) sludge in single-stage anaerobic digestion

Biohythane production via single-stage anaerobic digestion (AD) is an effective way for sustainable energy recovery from lignocellulosic biomass. In this paper, biohythane was produced through the AD process from pineapple peel waste substrate using purely cultured Methanosarcina mazei with the enhancement of palm oil mill effluent (POME) sludge as the inoculum. This study focuses on the effects of the lignocellulosic pre-treatment method, the addition of POME sludge into M. mazei culture medium as inoculum, and various operational conditions (food to microorganisms (F/M) ratios, temperature, pH) on gas production performances. The experimental results indicate that these parameters influenced the efficiency of biohythane production by producing the peak maximum biohythane production rate values (HPR_max) and (MPR_max), H₂:CH₄ = 1.93:0.67 L/L-d, and biohythane yield (HY) and (MY), H₂:CH₄ = 1.18:0.55 mL/L-substrate. This study demonstrates that biohythane gas (H₂ + CH₄ + CO₂) production from pineapple waste can be accelerated by M. mazei only with the enhancement of POME sludge through single-stage AD system under mesophilic batch process conditions.     

Alteration in Rhizosphere Soil Properties of Afforested <Emphasis Type="Italic">Rhamnus lycioides</Emphasis> Seedlings in Short-Term Response to Mycorrhizal Inoculation with <Emphasis Type="Italic">Glomus intraradices</Emphasis> and Organic Amendment

The reestablisment of autochthonous plant species is an essential strategy for recovering degraded areas under semiarid conditions. A field experiment was carried out to assess the short-term effect of two reafforestation methods involving mycorrhizal inoculation and compost addition on soil quality parameters and Rhamnus lycioides seedling growth. The nutrient content (NPK) and enzymatic activities (dehydrogenase, urease, protease-BAA, acid phosphatase and β-glucosidase) increased and bulk density decreased in the rhizosphere soil with the organic amendment. Biomass C of rhizosphere soil increased by at least 240% with respect to the control soil after mycorrhizal inoculation and the combination of compost addition + mycorrhizal inoculation. Both mycorrhizal inoculation and composted organic residue addition increased R. lycioides seedling growth in the same proportion. In the short term, we conclude that the application of both reafforestation methods not only enhances the establishment of R. lycioides seedlings, but also improves soil quality.     

Phytoremediation of heavy metals by <Emphasis Type="Italic">Eichhornia crassipes</Emphasis>

Eichhornia crassipes was tested for its ability to bioconcentrate 8 toxic metals (Ag, Cd, Cr, Cu, Hg, Ni, Pb, and Zn) commonly found in wastewater from industries. Young plants of equal size were grown hydroponically and amended with 0, 0.1, 0.3, 0.5, 1.0, 3.0, and 5.0 mM of each heavy metal individually for 21 days. The test plant had the lowest and the highest tolerance indices for Hg and Zn, respectively. A significant (P ≤ .05) reduction in biomass production was observed in metal treated plants compared with the control. All strace elements accumulated to higher concentrations in roots than in shoots. Trace element concentrations in tissues and the bioconcentration factors (BCF) were proportional to the initial concentration of individual metal in the growth medium and the duration of exposure. From a phytoremediation perspective, E. crassipes is a promising plant species for remediation of natural water bodies and/or wastewater polluted with low levels of Zn, Cr, Cu, Cd, Pb, Ag and Ni.     

接种根瘤菌和菌根真菌对三叶草生长及对土壤Pb2+吸收的影响

采用盆栽实验,设计了不接种(CK)、接种根瘤菌、接种丛枝菌根真菌、双接种根瘤菌和菌根真菌4种处理,探讨了三叶草的生长情况及对土壤中Pb2+吸收的影响。结果表明:三叶草对Pb具有一定的吸收能力,两次采样中,CK处理三叶草Pb浓度分别为15.38mg/kg和15.57mg/kg,均为各处理中最高;接种根瘤菌可明显提高三叶草生物量,接种菌根真菌能明显减少三叶草对土壤中Pb的吸收。接种根瘤菌和菌根真菌减少三叶草对Pb2+的吸收和向地上部运输。     

Enhancing phytoextraction: the effect of chemical soil manipulation on mobility, plant accumulation, and leaching of heavy metals

For heavy metal-contaminated agricultural land, low-cost, plant-based phytoextraction measures can be a key element for a new land management strategy. When agents are applied into the soil, the solubility of heavy metals and their subsequent accumulation by plants can be increased, and, therefore, phytoextraction enhanced. An overview is given of the state of the art of enhancing heavy metal solubility in soils, increasing the heavy metal accumulation of several high-biomass-yielding and metal-tolerant plants, and the effect of these measures on the risk of heavy metal leaching. Several organic as well as inorganic agents can effectively and specifically increase solubility and, therefore, accumulation of heavy metals by several plant species. Crops like willow (Salix viminalis L.), Indian mustard [Brassica juncea (L.) Czern.], corn (Zea mays L.), and sunflower (Helianthus annuus L.) show high tolerance to heavy metals and are, therefore, to a certain extent able to use the surpluses that originate from soil manipulation. More than 100-fold increases of lead concentrations in the biomass of crops were reported, when ethylenediaminetetraacetic acid (EDTA) was applied to contaminated soils. Uranium concentrations could be strongly increased when citric acid was applied. Cadmium and zinc concentrations could be enhanced by inorganic agents like elemental sulfur or ammonium sulfate. However, leaching of heavy metals due to increased mobility in soils cannot be excluded. Thus, implementation on the field scale must consider measures to minimize leaching. So, the application of more than 1 g EDTA kg(-1) becomes inefficient as lead concentration in crops is not enhanced and leaching rate increases. Moreover, for large-scale applications, agricultural measures as placement of agents, dosage splitting, the kind and amount of agents applied, and the soil properties are important factors governing plant growth, heavy metal concentrations, and leaching rates. Effective prevention of leaching, breeding of new plant material, and use of the contaminated biomass (e.g., as biofuels) will be crucial for the acceptance and the economic breakthrough of enhanced phytoextraction.     

Plant and Soil Responses to High and Low Diversity Grassland Restoration Practices

The USDA’s Conservation Reserve Program (CRP) has predominantly used only a few species of dominant prairie grasses (CP2 practice) to reduce soil erosion, but recently has offered a higher diversity planting practice (CP25) to increase grassland habitat quality. We quantified plant community composition in CP25 and CP2 plantings restored for 4 or 8 years and compared belowground properties and processes among restorations and continuously cultivated soils in southeastern Nebraska, USA. Relative to cultivated soils, restoration increased soil microbial biomass (P = 0.033), specifically fungi (P < 0.001), and restored soils exhibited higher rates of carbon (C) mineralization (P = 0.010). High and low diversity plantings had equally diverse plant communities; however, CP25 plantings had greater frequency of cool-season (C₃) grasses (P = 0.007). Older (8 year) high diversity restorations contained lower microbial biomass (P = 0.026), arbuscular mycorrhizal fungi (AMF) biomass (P = 0.003), and C mineralization rates (P = 0.028) relative to 8 year low diversity restorations; older plantings had greater root biomass than 4 year plantings in all restorations (P = 0.001). Low diversity 8 year plantings contained wider root C:N ratios, and higher soil microbial biomass, microbial community richness, AMF biomass, and C mineralization rate relative to 4 year restorations (P < 0.050). Net N mineralization and nitrification rates were lower in 8 year than 4 year high diversity plantings (P = 0.005). We attributed changes in soil C and N pools and fluxes to increased AMF associated with C₄ grasses in low diversity plantings. Thus, reduced recovery of AMF in high diversity plantings restricted restoration of belowground microbial diversity and microbially-mediated soil processes over time.     

Role of mycorrhizal fungi and phosphorus in the arsenic tolerance of basin wildrye

Revegetation of arsenic (As)-rich mine spoils is often impeded by the lack of plant species tolerant of high As concentrations and low nutrient availability. Basin wildrye [Leymus cinereus (Scribner & Merr.) A. L?ve] has been observed to establish naturally in soils with elevated As content and thus may be useful for the stabilization of As-contaminated soils. An experiment was conducted to evaluate how variable phosphorus (P) concentrations and inoculation with site-specific arbuscular mycorrhizal fungi influence As tolerance of basin wildrye. Basin wildrye was grown in sterile sand in the greenhouse for 16 weeks. Pots of sterile sand were amended to create one of four rates of As (0, 3, 15, or 50 mg As kg(-1)), two rates of P (3 or 15 mg P kg(-1)), and +/-mycorrhizal inoculation in a 2 x 4 x 2 factorial arrangement. After 16 weeks of growth, plants were harvested, shoots and roots thoroughly washed, and the tissue analyzed for total shoot biomass, total root and shoot As and P concentrations, and degree of mycorrhizal infection. Basin wildrye was found to be tolerant of high As concentrations allowing for vigorous plant growth at application levels of 3 or 15 mg As kg(-1). Arsenic was sequestered in the roots, with 30 to 50 times more As in the roots than shoots under low P conditions. Mycorrhizal infection did not confer As tolerance in basin wildrye nor did mycorrhizal fungi influence biomass production. Phosphorus concentrations of 15 mg kg(-1) effectively inhibited As accumulation in basin wildrye. Basin wildrye has the potential to be used for stabilization of As-rich soils while minimizing exposure to grazing animals following reclamation.     

Economic assessment of flash co-pyrolysis of short rotation coppice and biopolymer waste streams

The disposal problem associated with phytoextraction of farmland polluted with heavy metals by means of willow requires a biomass conversion technique which meets both ecological and economical needs. Combustion and gasification of willow require special and costly flue gas treatment to avoid re-emission of the metals in the atmosphere, whereas flash pyrolysis mainly results in the production of (almost) metal free bio-oil with a relatively high water content. Flash co-pyrolysis of biomass and waste of biopolymers synergistically improves the characteristics of the pyrolysis process: e.g. reduction of the water content of the bio-oil, more bio-oil and less char production and an increase of the HHV of the oil. This research paper investigates the economic consequences of the synergistic effects of flash co-pyrolysis of 1:1 w/w ratio blends of willow and different biopolymer waste streams via cost-benefit analysis and Monte Carlo simulations taking into account uncertainties. In all cases economic opportunities of flash co-pyrolysis of biomass with biopolymer waste are improved compared to flash pyrolysis of pure willow. Of all the biopolymers under investigation, polyhydroxybutyrate (PHB) is the most promising, followed by Eastar, Biopearls, potato starch, polylactic acid (PLA), corn starch and Solanyl in order of decreasing profits. Taking into account uncertainties, flash co-pyrolysis is expected to be cheaper than composting biopolymer waste streams, except for corn starch. If uncertainty increases, composting also becomes more interesting than flash co-pyrolysis for waste of Solanyl. If the investment expenditure is 15% higher in practice than estimated, the preference for flash co-pyrolysis compared to composting biopolymer waste becomes less clear. Only when the system of green current certificates is dismissed, composting clearly is a much cheaper processing technique for disposing of biopolymer waste.     

Fungal inoculum properties: extracellular enzyme expression and pentachlorophenol removal by New Zealand trametes species in contaminated field soils

This study was conducted to improve the ability of indigenous New Zealand white-rot fungi to remove pentachlorophenol (PCP) from contaminated field soil. The effects of different bioaugmentation conditions on PCP removal and extracellular enzyme expression were measured in the laboratory. The conditions were fungal growth substrate and co-substrate composition, culture age, and Tween 80 addition to the contaminated soil. The fungi used were Trametes versicolor isolate HR131 and Trametes sp. isolate HR577. Maximum PCP removal was 70% after 7 wk from a 1043 mg kg(-1) PCP-contaminated soil inoculated with an 11-d-old fungal culture of T. versicolor isolate HR131. There was minimal production of undesirable pentachloroanisole by the fungi. Tween 80 addition had no affect on PCP removal. Poplar sawdust was more suitable as a fungal growth substrate and a co-substrate amendment for PCP removal and extracellular enzyme expression than the locally available pine and fir sawdust. Pentachlorophenol removal was not necessarily correlated with extracellular enzyme expression. The research results demonstrate that PCP biodegradation was affected by inoculum culture age, by the presence of a co-substrate amendment, and by growth substrate composition after white-rot fungal bioaugmentation into PCP-contaminated field soils.     

Growth and fecundity of earthworms: <Emphasis Type="Italic">Perionyx excavatus</Emphasis> and <Emphasis Type="Italic">Perionyx sansibaricus</Emphasis> in cattle waste solids

Epigeic earthworms (Oligochaeta) have been appeared as key organisms to convert organic waste resources into value-added products, i.e., vermicompost and worm biomass. The assessment of reproduction potential of composting earthworm may be beneficial for large-scale earthworm production. Although, the waste minimizing potential of Perionyx excavatus and Perionyx sansibaricus is well proved, but little information is available about their fecundity rate. In this study, the efforts have been made to explore the growth and reproduction biology of P. excavatus and P. sansibaricus, using cattle waste solid as culture substrate, under laboratory conditions. Earthworms were weighed weekly and number of cocoons produced per week assessed. Biomass productions, fecundity, maturation, natality all were significantly different between P. excavatus and P. sansibaricus. The highest mean individual biomass was 767.7 ± 18.4 mg and 612.6 ± 20.6 mg, respectively in P. sansibaricus and P. excavatus. However, the highest cocoon numbers occurred in P. excavatus (492.3 ± 13.6), significantly higher than P. sansibaricus (269.6 ± 17.1). Fecundity was slightly different in both species: 1.38 ± 0.77 cocoons adult worm⁻¹ week⁻¹ (P. excavatus) and 1.58 ± 0.74 cocoons adult worm⁻¹ week⁻¹ (P. sansibaricus). The hatchling success rate (%) was highest in P. excavatus. Overall natality (juveniles adult⁻¹ week⁻¹) was highest in P. sansibaricus (1.52) than P. excavatus (1.26), which suggests that P. sansibaricus may be a better candidate for rapid propagation of earthworms in cattle waste solid.     

Comparison of vesicular-arbuscular mycorrhizae in plants from disturbed and adjacent undisturbed regions of a coastal salt marsh in Clinton,Connecticut, USA

Roots of salt marsh plant speciesSpartina alterniflora, S. patens, Distichlis spicata, and others were examined for the presence of vesicular-arbuscular mycorrhizal (VAM) fungi. Samples were taken from introduced planted material in a salt marsh restoration project and from native material in adjacent marsh areas along the Indian River, Clinton, Connecticut, USA. After ten years the replanted area still has sites devoid of vegetation. The salt marsh plants introduced there were devoid of VAM fungi, while high marsh species from the adjacent undisturbed region showed consistent infection, leading the authors to suggest that VAM fungal infection of planting stocks may be a factor in the success of marsh restoration.     

Comparison of Four <Emphasis Type="Italic">Sesbania</Emphasis> Species to Remediate Pb/Zn and Cu Mine Tailings

A 6-month greenhouse pot trial was performed, aimed at screening appropriate Sesbania species for remediation of Pb/Zn and Cu mine tailings. Performances of young seedlings of four Sesbania species (S. cannabina, S. grandiflora, S. rostrata, and S. sesban) were compared with and without inoculation of rhizobia. Seedlings were planted in two types of tailings amended with garden soil or garden soil mixed with river sediment. The results indicated that inoculated plants generally produced a higher biomass than samples without inoculation. Pb/Zn mine tailings containing rather high concentrations of total and water-soluble Cu, Pb, and Zn were toxic to plant growth compared with Cu mine tailings, according to the growth performance of the four species. Sesbania sesban and S. rostrata showed superior growth performance, compared to the other two species. Thus, they can serve as pioneer species to modify the barren environment, by providing organic matter and essential nutrients such as nitrogen, upon decomposition, in a relatively short period of time. This is especially true for S. rostrata, which is an annual plant that forms both stem and root nodules. However, a longer-term field trial should be conducted to investigate if superior species can beneficially modify the habitat for the growth of subsequent plant communities.     

Competitive biomethanation using substrates in combination and by cross inoculation

Summary Quantitative studies on the biomethanation processes using a different biomass (goat dung, cow dung, buffalo dung, piggery waste, poultry waste and sewage) alone or in combination have been made. The dung samples have been found to be an efficient producer of biogas at a 1:2 dilution. Better yields of biogas are obtained in combination with other biomasses rather than when used alone. Judicious mixing of biomasses, however, is important. Competitive biomethanation of a biomass by other biomasses as a source for a wild population of microbes has been studied in vials using a cross-inoculation technique, i.e. using inoculum of one biomass on different sterile biomasses. The results show that the microbes are very specific and usually non-adaptive. Each inoculum outclasses others in using its natural biomass for methanation but reacts poorly when inoculated to other alien biomasses. Buffalo dung is to some extent adaptive in nature.Professor S.C. Lahiri is the senior author of this paper and he is ex-Head of the Department of Chemistry at the University of Kalyani where Nilanjan Chakravorty is a research fellow. Dr G.M. Sarkar is a senior lecturer in the Department of Botany at Ranaghat College, Ranaghat, Nadia, West Bengal, India.     

Impact of transgenic Bt maize residues on the mycotoxigenic plant pathogen Fusarium graminearum and the biocontrol agent Trichoderma atroviride

Transformation of maize with genes encoding for insecticidal crystal (Cry) proteins from Bacillus thuringiensis (Bt) could have an impact on the saprophytic survival of plant pathogens and their antagonists on crop residues. We assessed potential effects on the mycotoxin deoxynivalenol (DON)-producing wheat and maize pathogen Fusarium graminearum and on the biocontrol agent Trichoderma atroviride. Purified Cry1Ab protein caused no growth inhibition of these fungi on agar plates. Cry1Ab concentrations above levels common in Bt maize tissue stimulated the growth of F. graminearum. The fungi were also grown on gamma-radiation-sterilized leaf tissue of four Bt maize hybrids and their non transgenic isolines collected at maize maturity on a field trial in 2002 and 2003. Both fungi degraded the Cry1Ab protein in Bt maize tissue. Fungal biomass quantification with microsatellite-based polymerase chain reaction (PCR) assays revealed differential fungal growth on leaf tissue of different maize varieties but no consistent difference between corresponding Bt and non-Bt hybrids. Generally, year of maize tissue collection had a greater impact on biomass production than cultivar or Bt transformation. The mycotoxin DON levels observed in maize tissue experiments corresponded with patterns in F. graminearum biomass, indicating that Bt transformation has no impact on DON production. In addition to bioassays, maize leaf tissue was analyzed with a mass spectrometer-based electronic nose, generating fingerprints of volatile organic compounds. Chemical fingerprints of corresponding Bt and non-Bt leaf tissues differed only for those hybrid pairs that caused differential fungal biomass production in the bioassays. Our results suggest that Cry1Ab protein in maize residues has no direct effect on F. graminearum and T. atroviride but some corresponding Bt/non-Bt maize hybrids differ more in composition than Cry protein content alone, which can affect the saprophytic growth of fungi on crop residues.     

Improved understanding of hyperaccumulation yields commercial phytoextraction and phytomining technologies

This paper reviews progress in phytoextraction of soil elements and illustrates the key role of hyperaccumulator plant species in useful phytoextraction technologies. Much research has focused on elements which are not practically phytoextracted (Pb); on addition of chelating agents which cause unacceptable contaminant leaching and are cost prohibitive; and on plant species which offer no useful phytoextraction capability (e.g., Brassica juncea Czern). Nickel phytoextraction by Alyssum hyperaccumulator species, which have been developed into a commercial phytomining technology, is discussed in more detail. Nickel is ultimately accumulated in vacuoles of leaf epidermal cells which prevents metal toxicity and provides defense against some insect predators and plant diseases. Constitutive up-regulation of trans-membrane element transporters appears to be the key process that allows these plants to achieve hyperaccumulation. Cadmium phytoextraction is needed for rice soils contaminated by mine wastes and smelter emissions with 100-fold more soil Zn than Cd. Although many plant species can accumulate high levels of Cd in the absence of Zn, when Cd/Zn>100, only Thlaspi caerulescens from southern France has demonstrated the ability to phytoextract useful amounts of Cd. Production of element-enriched biomass with value as ore or fertilizer or improved food (Se) or feed supplement may offset costs of phytoextraction crop production. Transgenic phytoextraction plants have been achieved for Hg, but not for other elements. Although several researchers have been attempting to clone all genes required for effective hyperaccumulation of several elements, success appears years away; such demonstrations will be needed to prove we have identified all necessary processes in hyperaccumulation.     

Soil amendments, plant age, and intercropping impact p,p'-DDE bioavailability to Cucurbita pepo

Field experiments were conducted to optimize the phytoextraction of weathered p,p'-DDE (p,p'-dichlorodiphenyldichloroethylene) by Cucurbita subspecies. The effects of two soil amendments, mycorrhizae or a biosurfactant, on p,p'-DDE accumulation was determined. Also, p,p'-DDE uptake was assessed during plant growth (12, 26, 38, and 62 d), and cultivars that accumulate weathered p,p'-DDE were intercropped with cultivars known not to have that ability. Cucurbita pepo L. ssp. pepo accumulated large amounts of the contaminant, having stem bioconcentration factors, amounts of p,p'-DDE translocated, and contaminant phytoextraction that were 14, 9.9, and 5.0 times greater than C. pepo L. ssp. ovifera (L.) D.S. Decker, respectively. During 62 d, the stem BCF (bioconcentration factor) for p,p'-DDE in subspecies pepo remained constant and the total amount of contaminant accumulated was correlated with plant biomass (r(2) = 0.86). For subspecies ovifera, the stem BCF was highest at 12 d (1.5) but decreased to 0.39 by 62 d, and p,p'-DDE removal was not correlated with plant biomass. Mycorrhizal inoculation increased p,p'-DDE accumulation by both subspecies by an average 4.4 times. For subspecies pepo, mycorrhizae increased the percentage of contaminant extracted from 0.72 to 2.1%. Biosurfactant amendment also enhanced contaminant accumulation by both subspecies, although treatment reduced subspecies ovifera biomass by 60%. The biosurfactant had no effect on the biomass of subspecies pepo, increased the average contaminant concentration by 3.6-fold, and doubled the overall amount of p,p'-DDE removed from the soil. Soil amendments that enhance the mobility of weathered persistent organic pollutants will significantly increase the amount of contaminant phytoextraction by Cucurbita pepo.     

Evaluation of heavy metals accumulation by two emergent macrophytes from the polluted soil: an experimental study

The concentrations of copper (Cu) and lead (Pb) in, and the biomass of, the different parts of Persicaria glabra (Willd.) Gamez and Juncellus alopecuroides (Rottb.) C.B.Cl. were evaluated while grown in pots under laboratory conditions. Cu and Pb were added as sulphates (50, 100, 200, 400 mg/kg) to the pots. Heavy metal concentrations in the plants were measured by atomic absorption spectrometry. Results reveal that the biomass of J. alopecuroides (particularly roots) was higher than P. glabra, and that the growth tendency of macrophytes decreased with increasing heavy metal concentration in the soil, while in P. glabra, biomass went on increasing with the increase in copper concentration. Heavy metal accumulation in the roots was more than in aerial parts, and, therefore, barring two exceptions, the transfer factor of heavy metals from roots to aerial parts showed as less than 1, suggesting less transfer of heavy metals from roots to aerial parts. Thus, these macrophytes are efficient accumulators of trace elements, particularly J. alopecuroides, which can be recommended for biofiltration of heavy metals from contaminated soils.     

Effect of liming on the distribution of heavy metals in untreated industrial sewage sludge produced in Pakistan for the cultivation of <Emphasis Type="Italic">Sorghum bicolor</Emphasis> (L.)

The recycling of sewage sludge on agriculture land represents an alternative, advantageous, disposal of this waste material. The aim of the present study was to evaluate the feasibility of using industrial sewage sludge, produced in Pakistan, as a fertiliser. Agricultural soil amended with 25% (w/w) sewage sludge with or without lime treatment was used for growing a variety of sorghum (PARC-SS-1). The mobility of the heavy metals (HMs) (Cd, Cu, Cr, Ni, Pb and Zn) and metalloid (As) in the untreated industrial waste water sewage sludge (UIWS) samples were assessed by applying a modified BCR (Community Bureau of Reference) sequential extraction procedure. The single extraction procedure comprised of the application of mild extractant (CaCl₂) and water for the estimation of the proportion of easily soluble metal fractions. The precision and accuracy of BCR was evaluated by using a certified reference material of soil amended with sewage sludge BCR 483. The plant available metal contents, as extracted by the deionised water and 0.01 M CaCl₂ solution and exchangeable fraction of BCR sequential, decreased with lime application in the range of 10–44% for As, Cr, Ni, Pb and Zn, except in the cases of Cd and Cu, where their mobility was increased by 10% and 24%, respectively. Sludge amendment enhanced the dry weight yield of sorghum and the increase was more obvious after liming up to 25%. The uptake of HMs were lower in test samples (3.2–21.8%), except for Cu and Cd, which was higher (4%), while they were below the permissible limit of these metals. The present experiment demonstrates that liming was important in factors facilitating the growth of sorghum in sludge-amended soil.