Phytoextraction of heavy metals by willows growing in biosolids under field conditions.

Biosolids produced by sewage treatment facilities can exceed guideline thresholds for contaminant elements. Phytoextraction is one technique with the potential to reduce these elements allowing reuse of the biosolids as a soil amendment. In this field trial, cuttings of seven species/cultivars of Salix(willows) were planted directly into soil and into biosolids to identify their suitability for decontaminating biosolids. Trees were irrigated and harvested each year for three consecutive years. Harvested biomass was weighed and analyzed for the contaminant elements: As, Cd, Cu, Cr, Hg, Pb, Ni, and Zn. All Salix cultivars, except S. chilensis, growing in soils produced 10 to 20 t ha(-1) of biomass, whereas most Salix cultivars growing in biosolids produced significantly less biomass (<6 t ha(-1)). Salix matsudana (30 t ha(-1)) and S. × reichardtii A. Kerner (18 t ha(-1)) had similar aboveground biomass production in both soil and biosolids. These were also the most successful cultivars in extracting metals from biosolids, driven by superior biomass increases and not high tissue concentrations. The willows were effectual in extracting the most soluble/exchangeable metals (Cd, 0.18; Ni, 0.40; and Zn, 11.66 kg ha(-1)), whereas Cr and Cu were extracted to a lesser degree (0.02 and 0.11 kg ha(-1)). Low bioavailable elements, As, Hg, and Pb, were not detectable in any of the aboveground biomass of the willows.

Simultaneous determination of organic acids and nitrate in xylem saps of the hyperaccumulator Alyssum murale by RP-HPLC after solid-phase extraction with nanosized hydroxyapatite.

An analytical approach based on solid-phase extraction (SPE) with nanosized hydroxyapatite and reversed-phase high performance liquid chromatography (RP-HPLC) for the simultaneous determination of organic acids (oxalic, malic, malonic, citric, and fumaric) and nitrate in xylem saps of the hyperaccumulator Alyssum murale is presented in this study. The optimum experimental conditions for the separation and determination of organic acids and nitrate are studied. The interference from nitrate on oxalic acid in RP-HPLC is eliminated by SPE with nanosized hydroxyapatite, and the simultaneous determination of organic acids and nitrate is achieved by RP-HPLC at the optimum chromatographic conditions. The accuracy of the method is confirmed with an average recovery ranging between 95.2% and 99.8%, the relative standard deviations (RSD) are less than 2.0%. This method is successfully applied to determine the organic acids and nitrate in xylem saps of the hyperaccumulator Alyssum murale.

Tolerance and accumulation of lead in Vetiveria zizanioides and its effect on oil production.

Experiments were conducted to evaluate lead tolerance and accumulation in vetiver grass Vetiveria zizanioides (L.), grown in hydroponics and a pot study and to examine the effect of lead on vetiver oil production. Elevated concentrations of lead decreased the length of shoots and roots of plants. However, vetiver grown in highly contaminated soils showed no apparent phytotoxicity symptoms. Lead concentrations in the shoots and roots of vetiver plants grown in hydroponics were up to 144 and 19530 mg kg(-1) and those grown in soil were 38 and 629 mg kg(-1), respectively. Lead had an effect on vetiver oil production and composition by stimulating oil yield and the number of its constituents. Oil yield ranged from 0.4-1.3%; the highest yields were found in plants grown in nutrient solution with 100 mg Pb l(-1) for 5 weeks (1.29%) and 7 weeks (1.22%). The number of total constituents of vetiver oil also varied between 47-143 compounds when lead was presentin the growth medium. The highest number (143) was found in plants grown in soil spiked with 1000 mg Pb kg(-1). The predominant compound was khusimol (10.7-18.1%) followed by (E)-isovalencenol (10.3-15.6%). Our results indicated that lead could increase the oil production of vetiver.

Cadmium tolerance and accumulation in cultivars of a high-biomass tropical tree (Averrhoa carambola) and its potential for phytoextraction.

Averrhoa carambola is a high-biomass tropical tree that has been identified as a Cd accumulator. In the present study, field survey, pot, and hydroponic experiments were conducted to investigate the variation of Cd tolerance and accumulation in cultivars of A. carambola as well as its potential for phytoextraction. In the field survey, it was found that concentrations of Cd in aerial tissues of A. carambola varied greatly among sites and cultivars. The Cd bioconcentration factors (BCFs) and Cd removals by the field-grown A. carambola differed significantly among sites but not among cultivars. Nonetheless, all four carambola cultivars investigated were able to accumulate considerably high concentrations of Cd in their shoots, which indicated that the 4-yr-old carambola stands could remove 0.3 to 51.8% of the total Cd content in the top 20-cm soil layer. When cultured in Cd-spiked soils, the carambola cultivar Hua-Di always showed higher Cd tolerance than the other cultivars; however, this tendency was not confirmed by hydroponic experiment. The Cd BCFs of cultivar Thailand grown in soils with 6 and 12 mg Cd kg(-1) were highest among cultivars, whereas this trend was reversed at 120 mg Cd kg(-1) treatment. Nevertheless, the pot- and hydroponics-grown carambola cultivars generally showed higher capacities to tolerate and accumulate Cd, compared with the control species. The present results indicate that a strong ability to tolerate and accumulate Cd seems to be a trait at the species level in A. carambola, although some degree of variances in both Cd tolerance and accumulation exists among cultivars.

Characterization of foliar manganese (Mn) in Mn (hyper)accumulators using X-ray absorption spectroscopy.

Plant hyperaccumulation of the essential nutrient manganese (Mn) is a rare phenomenon most evident in the Western Pacific region, and differs from hyperaccumulation of other elements. Mn hyperaccumulators employ a variety of species-dependent spatial distribution patterns in sequestering excess foliar Mn, including primary sequestration in both nonphotosynthetic and photosynthetic tissues. This investigation employed synchrotron X-ray absorption spectroscopy (XAS) in a comparative study of Mn (hyper)accumulators, to elucidate in situ the chemical form(s) of foliar Mn in seven woody species from Australia, New Caledonia and Japan. Foliar Mn was found to predominate as Mn(II) in all samples, with strong evidence of the role of carboxylic acids, such as malate or citrate, as complexing ligands. Overall, the X-ray absorption near-edge spectroscopy (XANES) and extended X-ray absorption fine-structure spectroscopy (EXAFS) data appeared weighted against previous observations that oxalate binds excess Mn in Mn-(hyper)accumulating species.

Hypotheses, mechanisms and trade-offs of tolerance and adaptation to serpentine soils: from species to ecosystem level.

Understanding the relative importance of the abiotic environment and species interactions in determining the distribution and abundance of organisms has been a challenge in ecological research. Serpentine substrata are stressful environments for plant growth due to multiple limitations, collectively called the "serpentine syndrome". In the present review, our aim is not only to describe recent work in serpentine ecology, but also to highlight specific mechanisms of species tolerance and adaptation to serpentine soils and their effects on community structure and ecosystem functioning. We present hypotheses of the development of serpentine endemism and a description of functional traits of serpentine plants together with a synthesis of species interactions in serpentine soils and their effects on community structure and ecosystem productivity. In addition, we propose hypotheses about the effects of the 'serpentine syndrome' on ecosystem processes including productivity and decomposition.

The effect of nitrogen form on rhizosphere soil pH and zinc phytoextraction by Thlaspi caerulescens.

The phytoextraction of Zn may be improved by applying N fertilizers to increase the biomass and Zn content of shoots. Rhizosphere-pH change from uptake of different N forms will affect Zn phyto-availability in the rhizosphere and Zn phytoextraction. This glasshouse study examined the effect of N form on Zn phytoextraction by Thlaspi caerulescens (Prayon). The plants were grown in a Zn-contaminated soil (total Zn 250 mg kg-1 soil; pHwater 5.7) and supplied with (NH4)2SO4, Ca(NO3)2 or urea [(NH2)2CO]. The form was maintained by applying the nitrification inhibitor dicyandiamide. A biodegradable chelator ethylenediaminedisuccinic acid (EDDS) was included for comparison. The addition of N doubled the shoot biomass. The highest shoot Zn content occurred in the Ca(NO3)2 treatment and was associated with the highest rhizosphere pH. The lowest shoot dry weight occurred in the EDDS treatment. The Zn concentration in the shoots increased as the rhizosphere pH increased. A significant correlation occurred between Ca and Zn concentrations in the shoots. This study demonstrated that Ca(NO3)2 is a more effective treatment than , urea or EDDS for enhancing Zn phytoextraction in a mildly acidic soil.

Effects of phytoextraction on heavy metal concentrations and pH of pore-water of biosolids determined using an in situ sampling technique.

Heavy metal concentrations and pH of pore-water in contaminated substrates are important factors in controlling metal uptake by plants. We investigated the effects of phytoextraction on these properties in the solution phase of biosolids and diluted biosolids in a 12-month phytoextraction column experiment. Phytoextraction using Salix and Populus spp. temporarily decreased pore-water pH of the substrates over the experimental period followed by a return to initial pH conditions. Salixxreichardtii and Populus balsamifera effectively extracted Ni, Zn and Cd and actively mobilized these metals from the solid to the solution phase. S.xreichardtii had the stronger effect on mobilization of metals due to its larger root system. Phytoextraction did not affect Cu in the solution phase of the biosolids. Heavy metals were leached down to lower depths of the columns during the phytoextraction process.

Foliar manganese accumulation by Maytenus founieri (Celastraceae) in its native New Caledonian habitats: populational variation and localization by X-ray microanalysis.

Hyperaccumulation by plants is a rare phenomenon that has potential practical benefits. The majority of manganese (Mn) hyperaccumulators discovered to date occur in New Caledonia, and little is known about their ecophysiology. This study reports on natural populations of one such species, the endemic shrub Maytenus founieri. Mean foliar Mn concentrations of two populations growing on ultramafic substrates with varying soil pHs were obtained. Leaf anatomies were examined by light microscopy, while the spatial distributions of foliar Mn in both populations were examined by qualitative scanning electron microscopy/energy dispersive spectroscopy (SEM/EDS). Plants growing on two different substrates were found to have very different mean dry weight (DW) foliar Mn concentrations. Light microscopy showed that the leaves had very distinct thick dermal structures, consisting of multiple layers of large cells in the hypodermis. In vivo X-ray microprobe analyses revealed that, in both populations, Mn sequestration occurred primarily in these dermal tissues. The finding here that foliar Mn is most highly localized in the nonphotosynthetic tissues of M. founieri contrasts with results from similar studies on other woody species that accumulate high Mn concentrations in their shoots.

Uptake and accumulation of arsenic by 11 Pteris taxa from southern China.

A field survey was conducted at a deserted arsenic (As) mine in Guangxi Province, China to explore new potential As hyperaccumulators. In addition, young plants of 11 Pteris taxa were grown in glasshouse conditions for 12 weeks on As-amended soils with 0, 50 and 200 mg As kg(-1). Results of the field survey showed that the fern Pteris fauriei accumulated over 1000 mg As kg(-1) in its fronds. Of the 11 Pteris taxa, Pteris aspericaulis, Pteris cretica var. nervosa, P. fauriei, Pteris multifida, P. multifida f. serrulata, and Pteris oshimensis were all found to hyperaccumulate As in addition to P. cretica 'Albo-Lineata' and Pteris vittata (already reported as As hyperaccumulators). However, Pteris ensiformis, Pteris semipinnata and Pteris setuloso-costulata showed no evidence of As hyperaccumulation. Results also revealed a constitutive property of As hyperaccumulation in different populations of P. cretica var. nervosa, P. multifida, P. oshimensis and P. vittata.

Nickel and other metal uptake and accumulation by species of Alyssum (Brassicaceae) from the ultramafics of Iran.

Some plants growing on serpentine (ultramafic) soils are able to hyperaccumulate nickel in their above-ground parts. The genus Alyssum L. contains the greatest number of Ni-hyperaccumulator plants so far reported. There are substantial areas of serpentine soils at many locations in Iran. This paper presents the analyses for Ni, Cr, Mn, Fe, Mg and Ca in soils and Alyssum species from the ultramafics of west and northwest Iran. Soil analysis for total elements in these areas indicates that typical concentrations of Ni, Cr, Mn, Fe, Mg and Ca are up to about 1240, 365, 800, 51,150, 152,390 and 11,790 microg g(-1), respectively. During this study, seven Alyssum species were collected. Analysis of leaf dry matter shows that Alyssum bracteatum can contain up to 2300 microg Nig(-1), while the other species contain much lower concentrations of Ni and other elements. A. bracteatum is endemic to Iran and the first Ni hyperaccumulator reported from this species.

Growth and lead accumulation by the grasses Vetiveria zizanioides and Thysanolaena maxima in lead-contaminated soil amended with pig manure and fertilizer: a glasshouse study.

Bo Ngam lead mine soils contain high concentrations of lead (up 1% total Pb) and low amounts of organic matter and major nutrients (N, P, K). A glasshouse study was conducted to compare growth performance, metal tolerance and metal uptake by two grasses, Thysanolaena maxima (Roxb.) O. Kuntze and four ecotypes of Vetiveria zizanioides (L.) Nash, syn. Chrysopogon zizanioides (L.) Roberty (three from Thailand: Surat Thani, Songkhla and Kamphaeng Phet, and one from Sri Lanka) and to study the effects of pig manure (20% and 40% w/w) and inorganic fertilizer (75 and 150 mg kg(-1)) amendments to this lead mine soil. The results showed that both T. maxima and V. zizanioides (Surat Thani and Songkhla) could tolerate high Pb concentrations in soil (10750 mg kg(-1)) and had very good growth performance. Application of pig manure increased electrical conductivity (EC) and reduced DTPA-extractable Pb concentration in the soils. Pig manure application improved the growth of vetiver, especially at 20%, application dosage. Vetiver had the highest biomass. T. maxima could not tolerate high EC values. The uptake by roots and transport of Pb to shoots of both species was reduced when soils were amended with pig manure. Application of inorganic fertilizer did not improve growth of vetiver but did improve that of T. maxima. Fertilizer application did not have any great influence on the Pb uptake in vetiver while T. maxima took up more Pb as a result of the fertilizer enhancing its biomass yield. Both species transported low Pb concentrations to shoots (8.3-179 mg kg(-1)) and accumulated higher concentrations in roots (107-911 mg kg(-1)). In summary, both species may be species well suited for phytostabilization in tropical lead mine areas.

Manganese accumulation in the leaf mesophyll of four tree species: a PIXE/EDAX localization study.

Little is known about the spatial distribution of excess manganese (Mn) in the leaves of tolerant plants. Recently, the first such study of a Mn hyperaccumulator showed that the highest localized Mn concentrations occur in the photosynthetic tissue. This is in contrast to reports based on localization of foliar accumulation of other heavy metals. Here, four tree species, Gossia bidwillii, Virotia neurophylla, Macadamia integrifolia and Macadamia tetraphylla, which hyperaccumulate or strongly accumulate Mn, were studied. Cross-sectional foliar Mn localization was carried out in situ using proton-induced X-ray emission/energy dispersive X-ray analysis (PIXE/EDAX). All four species contained photosynthetic tissues with multiple palisade layers. These were shown to be the primary sequestration sites for Mn. Mn was not detected in the epidermal tissues. The findings of this study demonstrate a concurrence of three traits in four tree species, that is, accumulation of excess Mn in the leaves, its primary sequestration in the photosynthetic tissues, and multiple-layer palisade mesophyll.

Cellular and subcellular compartmentation of Ni in the Eurasian serpentine plants Alyssum bracteatum, Alyssum murale (Brassicaceae) and Cleome heratensis (Capparaceae).

This study investigated the cellular and subcellular compartmentation of Ni in the Eurasian serpentine species Alyssum murale, Alyssum bracteatum and Cleome heratensis and a non-serpentine population of A. murale (as a control) grown in hydroponic culture. Plant growth responses and Ni uptake clearly revealed the higher Ni tolerance of serpentine plants than the non-serpentine plants. Serpentine A. murale and A. bracteatum grew better at elevated (0.01 mM) Ni in the nutrient solution, supporting the view that the Ni hyperaccumulators have a higher requirement for Ni than normal plants. Low shoot Ni content of C. heratensis in response to the high Ni treatments indicated that this species employs an avoidance strategy for Ni tolerance. Energy-dispersive X-ray microanalysis showed that Ni was highly concentrated in the cell walls and cell lumen, most likely the vacuoles, of leaf epidermis of A. murale and A. bracteatum rather than in the mesophyll cells. EDX spectra from leaves of the non-serpentine A. murale suggested that Ni accumulated in both epidermal and mesophyll cells but not in the epidermal cell walls. Growth reduction and Ni toxicity in plants of the non-serpentine A. murale could be due to accumulation of Ni in the lumen of leaf mesophyll cells. Our data suggest that cellular and subcellular compartmentation are both possible mechanisms for Ni tolerance employed by the serpentine A. murale and A. bracteatum.

Uptake and accumulation of lead by plants from the Bo Ngam lead mine area in Thailand.

A field survey of terrestrial plants growing on Bo Ngam lead mine area, Thailand, was conducted to identify species accumulating exceptionally high concentrations of lead. Plant and soil samples were collected from five areas. Lead concentrations in surface soil ranged from 325 to 142,400 mg/kg. The highest lead concentration in soil was found at the ore dressing plant area and lowest at a natural pond area. In different areas, the concentrations of lead in plants were different when comparing various study sites. A total of 48 plant species belonging to 14 families were collected from five sampling sites. Twenty-six plant species had lead concentrations more than 1000 mg/kg in their shoots. Three species (Microstegium ciliatum, Polygala umbonata, Spermacoce mauritiana) showed extremely high lead concentrations in their shoots (12,200-28,370 mg/kg) and roots (14,580-128,830 mg/kg).

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.

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.

Copper tolerance, uptake and accumulation by Phragmites australis.

Copper (Cu) uptake and accumulation of five populations of Phragmites australis growing in two mine sites (Parys Mountain, Wales, UK and Plombières, Belgium) contaminated with Cu and three 'clean' sites (Felixstowe, UK; Wisbech, UK and Mai Po, Hong Kong) were studied under field and glasshouse conditions. Cu tolerances in these populations of seedlings raised from seeds collected from the above five sites were also studied under glasshouse conditions. Although concentrations of Cu in the plant tissues (leaves, stems, rhizomes and roots) of P. australis and the associated soils from the Cu-contaminated sites were significantly higher than those of the plant tissues and the soils collected in the clean sites, small differences were found between the two Cu-contaminated populations and the three clean populations when seedlings were grown in 0.1 and 0.5 microg ml(-1) Cu treatment solutions. In general, different populations of seedlings showed similar growth responses, metal uptake and indices of Cu tolerance when cultured in the same Cu treatment solution for 3 weeks. There was insufficient evidence to support the hypothesis that the Cu-contaminated populations have evolved to Cu-tolerant ecotypes.

Soil Cd availability to Indian mustard and environmental risk following EDTA addition to Cd-contaminated soil.

A pot experiment was conducted to investigate the influence of EDTA on the extractability of Cd in the soil and uptake of Cd by Indian mustard (Brassica juncea). Twenty levels of soil Cd concentration ranging from 10 to 200 mg kg(-1) were produced by spiking aliquots of a clay loam paddy soil with Cd(NO3)2. One week before the plants were harvested EDTA was applied to pots in which the soil had been spiked with 20, 40, 60...200 mg Cd kg(-1). The EDTA was added at the rate calculated to complex with all of the Cd added at the 200 mg kg(-1) level. Control pots spiked with 10, 30, 50... 190 mg Cd kg(-1) received no EDTA. The plants were harvested after 42 days' growth. Soil water- and NH4NO3-extractable Cd fractions increased rapidly following EDTA application. Root Cd concentrations decreased after EDTA application, but shoot concentrations increased when the soil Cd levels were >130 mg kg(-1) and Cd toxicity symptoms were observed. The increases in soil solution Cd induced by EDTA did not increase plant total Cd uptake but appeared to stimulate the translocation of the metal from roots to shoots when the plants appeared to be under Cd toxicity stress. The results are discussed in relation to the possible mechanisms by which EDTA may change the solubility and bioavailability of Cd in the soil and the potential for plant uptake and environmental risk due to leaching losses to groundwater.

Evolutionary dynamics of nickel hyperaccumulation in Alyssum revealed by its nrDNA analysis.

• Molecular phylogeny based on ribosomal internal transcribed spacer (ITS) sequences was studied to investigate the phyletic relationships among some nickel (Ni)-hyperaccumulating and nonhyperaccumulating species of the genus Alyssum in relation to their geographic distribution and Ni-hyperaccumulating phenotype. • Thirty-seven samples belonging to 32 taxa were analysed by sequencing the polymerase chain reaction-amplified ITS region and performing neighbor joining, maximum parsimony and maximum likelihood phylogenetic analyses. • The ITS region in the sampled species varied from 221 to 307 bp of ITS1 and from 194 to 251 bp of ITS2. A total of 765 characters was used to infer the phylogeny and the average nucleotide variation detected was 15.15%. • Nickel-hyperaccumulation could have been lost or acquired independently more than once during the speciation of the genus. The geographical location of species could not be related to phylogenetic affinities.