Effect of simultaneous establishment of Sedum alfredii and Zea mays on heavy metal accumulation in plants

Land application of biosolids to improve agricultural productivity is a cost-effective approach for resource recovery. Unfortunately, municipal biosolids often contain high concentrations of heavy metals, including zinc and copper. In this study, a co-cropping technique was investigated using a known zinc hyperaccumulator, Sedum alfredii with a grain crop, Zea mays. After a 3-mo growth trial, the results indicate that when Z. mays is co-cropped with S. alfredii, heavy metals accumulated in the grains were significantly reduced when compared to monoculture cropping. Co-cropping improved the growth of both plant species. In addition, the biosolids maintained stable pH, N-P-K concentrations, germination potential, and water content after the plant treatment, regardless of the plant species used in the trial. In conclusion, co-cropping with hyperaccumulators may be an effective approach to reducing the risk of contaminant uptake in edible crops.     

Effects of bacteria on enhanced metal uptake of the Cd/Zn-hyperaccumulating plant, Sedum alfredii

To investigate the effects of bacteria (Burkholderia cepacia) on metal uptake by the hyperaccumulating plant, Sedum alfredii, a hydroponic experiment with different concentrations of Cd and Zn was conducted. It was found that inoculation of bacteria on S. alfredii significantly enhanced plant growth (up to 110% with Zn treatment), P (up to 56.1% with Cd treatment), and metal uptake (up to 243% and 96.3% with Cd and Zn treatment, respectively) in shoots, tolerance index (up to 134% with Zn added treatment), and better translocation of metals (up to 296% and 135% with Cd and Zn treatment, respectively) from root to shoot. In the ampicillin added treatment with metal addition, stimulation of organic acid production (up to an increase of 133% of tartaric acid with Cd treatment) by roots of S. alfredii was observed. The secretion of organic acids appears to be a functional metal resistance mechanism that chelates the metal ions extracellularly, reducing their uptake and subsequent impacts on root physiological processes.     

COLONIZATION AND MODULATION OF HOST GROWTH AND METAL UPTAKE BY ENDOPHYTIC BACTERIA OF SEDUM ALFREDII

Sedum alfredii Hance is a Zn and Cd co-hyperaccumulating plant species found in an old mining area in China. Four bacterial strains, Burkholderia sp. SaZR4, Burkholderia sp. SaMR10, Sphingomonas sp. SaMR12 and Variovorax sp. SaNR1, isolated from surface-sterilized S. alfredii plants were used to investigate their endophytic nature and root colonization patterns and effects on phytoextraction of Zn and Cd. Laser scanning confocal microscopy revealed that gfp-tagged SaZR4, SaMR12, and SaNR1 cells formed biofilms on roots and that SaZR4 and SaMR12 cells could invade root tissues. SaMR10 showed the lowest total population associated with S. alfredii and little effect on plant growth and phytoextraction. SaZR4 significantly promoted Zn-extraction but not Cd-extraction. SaMR12 and SaNR1 significantly promoted plant growth in substrates supplemented with Zn or Cd and phytoextraction of Zn and Cd. Together, this study have shown that the four native endophytic bacteria differently colonize the host plants and modulate metal uptake and growth of host plant, and that SaMR12 and SaNR1 strains are promising assistants of S. alfredii plants for phytoremediation of Zn/Cd-contaminated soil.     

Influence of Rapeseed Cake on Heavy Metal Uptake by a Subsequent Rice Crop After Phytoextraction Using Sedum plumbizincicola

A glasshouse pot experiment was conducted to study the effects of phytoextraction by Sedum plumbizincicola and application of rapeseed cake (RSC) on heavy metal accumulation by a subsequent rice (Oryza sativa L.) crop in a contaminated paddy soil collected from east China. After phytoextraction by S. plumbizincicola the soil and brown rice Cd concentrations effectively declined. After phytoextraction, RSC application reduced brown rice Cd concentrations in the subsequent rice crop to 0.23–0.28 mg kg^?1, almost down to the standard limit (0.2 mg kg^?1). After phytoextraction and then application of RSC, the soil solution pH, dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) concentrations increased during early stages of rice growth resulting directly and indirectly in lowering the bioavailability of the heavy metals. Thus the grain yield of the subsequent rice crop increased and the heavy metals in the brown rice declined significantly. In this contaminated acid soil, growing the hyperaccumulator S. plumbizincicola and rice in rotation together with RSC application may therefore be regarded as a viable strategy for safe grain production and bioremediation.     

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.     

Soil washing of Pb, Zn and Cd using biodegradable chelator and permeable barriers and induced phytoextraction by Cannabis sativa

The feasibility of combined phytoextraction and in situ washing of soil contaminated with Pb (1750 mg kg^–1), Zn (1300 mg kg^–1), and Cd (7.2 mg kg^–1), induced by the addition of biodegradable chelator, [S,S] stereoisomere of ethylenediamine discuccinate ([S,S]-EDDS), was tested in soil columns with hemp (Cannabis sativa) as the phytoextracting plant. The addition of [S,S]-EDDS (10 mmol kg^–1 dry soil) yielded concentrations of 1026±442, 330.3±114.7 and 3.84±1.55 mg kg^–1 of Pb, Zn and Cd in the dry above-ground plant biomass, respectively. These concentrations were 1926, 7.5, and 11 times higher, respectively, compared to treatments with no chelator addition. Horizontal permeable barriers, composed of a 3 cm high layer of nutrient enriched sawdust and vermiculite mixture, and a 3 cm layer of soil, vermiculite and apatite mixture, were positioned 20, 30 and 40 cm deep in the soil. In chelator treatments, barriers placed 30 cm deep reduced leaching of Pb, Zn and Cd by 435, 4 and 53 times, respectively, compared to columns with no barrier, where 3.0, 4.3 and 3.3% of total initial Pb, Zn and Cd, respectively, was leached during 6-weeks water irrigation after chelator addition. Lower positioned barriers were almost equally effective in preventing leaching of Pb than barriers positioned closer to the soil surface, less effective for Cd, and did not prevent leaching of Zn. 2.53% of total initial Pb and 2.83% of Cd was washed from the contaminated soil and accumulated into the barrier. Combined method was less effective than simulated ex situ soil washing, where 14.2, 5.5 and 24.5% of Pb, Zn and Cd, respectively, were removed after 1-h extraction, but comparable effective to 48-h extraction. Abbreviations: BCF – bioconcentration factor; EDTA – ethylene diaminetetraacetate; HM – heavy metal; PP – phytoextraction potential; [S,S]-EDDS – [S,S]-ethylenediamine disuccinate.     

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.     

Long-term field phytoextraction of zinc/cadmium contaminated soil by Sedum plumbizincicola under different agronomic strategies

In two long-term field experiments the zinc (Zn)/cadmium (Cd) hyperaccumulator Sedum plumbizincicola (S. plumbizincicola) was examined to optimize the phytoextraction of metal contaminated soil by two agronomic strategies of intercropping with maize (Zea mays) and plant densities. Soil total Zn and Cd concentrations decreased markedly after long-term phytoextraction. But shoot biomass and Cd and Zn concentrations showed no significant difference with increasing remediation time. In the intercropping experiment the phytoremediation efficiency in the treatment “S. plumbizincicola intercropped with maize” was higher than in S. plumbizincicola monocropping, and Cd concentrations of corn were below the maximum national limit. In the plant density experiment the phytoremediation efficiency increased with increasing plant density and 440,000 plants ha^?1 gave the maximum rate. These results indicated that S. plumbizincicola at an appropriate planting density and intercropped with maize can achieve high remediation efficiency to contaminated soil without affecting the cereal crop productivity. This cropping system combines adequate agricultural production with soil heavy metal phytoextraction.     

Identifying the functional groups and the influence of synthetic chelators on Cd availability and microbial biomass carbon in Cd-contaminated soil

Synthetic chelators play an important role in boosting the microbial biomass carbon (MBC), dissolved organic carbon (DOC), and heavy metal solubility in a contaminated soil toward a sustainability of environment for agricultural crops. Castor plant was grown under different levels of Cd contaminated soil (?Cd and +Cd) following adding three chelating agents, ethylenediaminetetraacetic acid (H₄EDTA), nitriloacetic acid (H₃ NTA), and NH₄ citrate (ammonium citrate) to the soil at rates of 10, 15, and 25 mmol in 5 kg of soil per pot. The highest bioavailable Cd concentrations in soil and castor plant were obtained from NH₄ citrate and H₄EDTA treatments in the contaminated soil. Fourier transform infrared (FTIR) analysis showed that NH₄ citrate was the most effective chelator in Cd-contaminated soil. MBC and DOC contents were significantly increased and reached at 81.98–80.37 and 1.96–1.90 mg kg^?1 respectively, in the (H₃ NTA) and NH₄ citrate treatments in Cd-contaminated soil. Further research is needed to investigate the use of chelators in the phytoextraction of Cd-contaminated soils under field conditions and whether it may be beneficial in accelerating the phytoextraction of Cd through hyperaccumulating plants.     

Contribution of Ectomycorrhizal Fungi to Cadmium Uptake of Poplars and Willows from a Heavily Polluted Soil

Phytoextraction has been proposed in recent years as an environmentally and cost-efficient treatment technique for the remediation of heavy-metal contaminated sites. In particular, plants that are fast growing, metal accumulating, and economically interesting, such as sunflowers or trees, recently became more important in research on phytoextraction. Heavy metal uptake of trees can be strongly influenced by ectomycorrhizal fungi. We investigated the possibility of enhancing phytoextraction of Cd by willows (Salix viminalis) and poplars (Populus canadensis) in association with three well known ectomycorrhizal fungi (Hebeloma crustuliniforme, Paxillus involutus and Pisolithus tinctorius). A pot experiment was conducted using Cd polluted soil from a contaminated site. Four replicates of each combination of fungus and tree species, and controls without fungal inoculum, were set up. After a growth period of 11 weeks, yields and Cd concentrations in roots, stems, and leaves were measured. In addition, the total Cd uptake, the transfer to roots, and the translocation to stems and leaves were calculated. The association of P. canadensis with P. involutus led to a highly significant increase of Cd concentrations, in particular in the leaves, which contained 2.74 ± 0.34 mg Cd per kg dry matter. Compared to the control this is an enhancement of nearly 100%. The fungi also significantly enhanced the translocation from the roots to the leaves, leading to a concentration ratio (leaves/roots) of 0.32 ± 0.06 compared to 0.20 ± 0.02 of the control plants. Additionally, P. involutus significantly enhanced the total Cd extraction by P. canadensis. Similar effects were not observed by other fungi or in association with S. viminalis.     

Remediation and Safe Production of cd Contaminated Soil Via Multiple Cropping Hyperaccumulator Solanum nigrum L. and Low Accumulation Chinese Cabbage

Multiple crop experiment of hyperaccumulator Solanum nigrum L. with low accumulation Chinese cabbage Fenyuanxin 3 were conducted in a cadmium (Cd) contaminated vegetable field. In the first round, the average removal rate of S. nigrum to Cd was about 10% without assisted phytoextraction reagent addition for the top soil (0–20 cm) with Cd concentration at 0.53–0.97 mg kg^?1 after its grew 90 days. As for assisted phytoextraction reagent added plots, efficiency of Cd remediation might reach at 20%. However, in the second round, Cd concentration in Chinese cabbage was edible, even in the plots with assisted phytoextraction reagent added. Thus, multiple cropping hyperaccumulator with low accumulation crop could normally remediate contaminated soil and produce crop (obtain economic benefit) in one year, which may be one practical pathway of phytoremediating heavy metal contaminated soil in the future.     

Co-planting can phytoextract similar amounts of cadmium and zinc to mono-cropping from contaminated soils

Co-planting crops normally decreases the main crop yield due to the reduced soil surface area occupied by the main crop. However, in our previous experiments, co-planting Sedum alfredii, a shade-requiring, Cd and Zn-hyperaccumulating plant, with corn increased the biomass and metal phytoextraction of S. alfredii. This experiment was conducted to verify if co-planting another hyperaccumulator, Thlaspi caerulescens, with ryegrass (Lolium perenne) in a pot-trial could obtain a similar result. The soil was separated by two permeable nets with a 2 mm interface soil layer to obtain a shared rhizosphere zone. Soluble metal concentrations in the soil in different rooting zones were measured using 0.01 mol L^?1 CaCl₂ extraction. The results showed that the growth of T. caerulescens was significantly promoted by co-planting, with a growth increase of about 2-fold compared with monoculture growth. The total uptake of Cd and Zn by T. caerulescens was not decreased by co-planting, and resulted in similar phytoextraction rates for Cd (about 26.6% of the soil total Cd) and Zn (about 2.4% of the soil total Zn) when compared with monoculture, though the T. caerulescens population was decreased by 50% because of co-planting. Analysis of soil samples showed that T. caerulescens substantially reduced the concentrations of 0.01 mol L^?1 CaCl₂ extractable Cd and Zn throughout the soil, even in the interface area and the ryegrass rooting area. The ryegrass roots did not mobilize more metals for the co-planted T. caerulescens. Based on these results, existing grass on contaminated land could be partly left while planting metal hyperaccumulators for phytoremediation in order to reduce runoff from the contaminated soil. However a field scale trial would be required for these results to be verified.     

Rhizosphere concentrations of zinc and cadmium in a metal contaminated soil after repeated phytoextraction by Sedum plumbizincicola

A growth chamber pot experiment and a field plot experiment were conducted with the installation of rhizobags to study the effects of repeated phytoextraction by Sedum plumbizincicola on the bioavailability of Cd and Zn in the rhizosphere and bulk soil Repeated phytoextraction gave significantly lower Cd and Zn concentrations in both rhizosphere and bulk soil solutions compared with soil without repeated phytoextraction. The depletion rates of NH40Ac-extractable Zn in rhizosphere soil in each treatment (L-PS, L-NPS, H-PS, and H-NPS) were 59.7, 18.0, 16.3, and 18.6%, respectively. For NH40Ac-extractable Cd, the depletion rates in treatments L-PS, L-NPS, H-PS, and H-NPS were 6.67, 29.4, 40.3, and 41.4%, respectively. Plant shoot biomass decreased in the order H-PS > H-NPS > L-PS > L-NPS, with dry weights of 0.56, 0.42, 1.43, and 1.21 g pot(-1), respectively. Plant Cd uptake increased with increasing aqua-regia extractable metal concentrations. The NH4OAc extraction procedure was satisfactory to predict the bioavailability of Cd and Zn in rhizosphere soil in terms of shoot uptake by S. plumbizincicola with positive correlation coefficients of 0.545 (p < 0.05) and 0.452 (p < 0.05), respectively. The field study results show a slight decrease in water soluble and NH4OAc-extractable metals, a trend similar to that found in the pot experiment.     

Chelator-assisted phytoextraction of arsenic,cadmium and lead by Pteris vittata L. and soil microbial community structure response

Abstract

Using biodegradable chelators to assist in phytoextraction may be an effective approach to enhance the heavy-metal remediation efficiencies of plants. A pot experiment was conducted to investigate the effects of ethylenediamine disuccinic acid (EDDS), citric acid (CA), and oxalic acid (OA) on the growth of the arsenic (As) hyperaccumulator Pteris vittata L., its arsenic (As), cadmium (Cd), and lead (Pb) uptake and accumulation, and soil microbial responses in multi-metal(loid)-contaminated soil. The addition of 2.5-mmol kg^?1 OA (OA-2.5) produced 26.7 and 14.9% more rhizoid and shoot biomass, respectively compared with the control, while EDDS and CA treatments significantly inhibited plant growth. The As accumulation in plants after the OA-2.5 treatment increased by 44.2% and the Cd and Pb accumulation in plants after a 1-mmol kg^?1 EDDS treatment increased by 24.5 and 19.6%, respectively. Soil urease enzyme activities in OA-2.5 treatment were significantly greater than those in the control and other chelator treatments (p?<?0.05). A PCR–denatured gradient gel electrophoresis analysis revealed that with the addition of EDDS, CA and OA enhanced soil microbial diversity. It was concluded that the addition of OA-2.5 was suitable for facilitating phytoremediation of soil As and did not have negative effects on the microbial community.     

Soil pH Effects on Uptake of Cd and Zn by Thlaspi caerulescens

For phytoextraction to be successful and viable in environmental remediation, strategies that can optimize plant uptake must be identified. Thlaspi caerulescens is an important hyperaccumulator of Cd and Zn, whether adjusting soil pH is an efficient way to enhance metal uptake by T. caerulescens must by clarified. This study used two soils differing in levels of Cd and Zn, which were adjusted to six different pH levels. Thlaspi caerulescens tissue metal concentrations and 0.1 M Sr(NO₃)₂ extractable soil metal concentrations were measured. The soluble metal form of both Cd and Zn was greatly increased with decreasing pH. Lowering pH significantly influenced plant metal uptake. For the high metal soil, highest plant biomass was at the lowest soil pH (4.74). The highest shoot metal concentration was at the second lowest pH (5.27). For low metal soil, due to low pH induced Al and Mn toxicity, both plant growth and metal uptake was greatest at intermediate pH levels. The extraordinary Cd phytoextraction ability of T. caerulescens was further demonstrated in this experiment. In the optimum pH treatments, Thlaspi caerulescens extracted 40% and 36% of total Cd in the low and high metal soils, respectively, with just one planting. Overall, decreasing pH is an effective strategy to enhance phytoextraction. But different soils had various responses to acidification treatment and a different optimum pH may exist. This pH should be identified to avoid unnecessarily extreme acidification of soils.     

Phytoextraction potential of soils highly polluted with cadmium using the cadmium/zinc hyperaccumulator Sedum plumbizincicola

Abstract

A three-crop repeated phytoextraction experiment was conducted using four soils (S1–S4) highly polluted with cadmium (Cd) and two enhanced phytoextraction pot experiments using the most polluted soil (S4) to investigate the feasibility of Cd removal from highly polluted soils using the Cd/zinc (Zn)-hyperaccumulator Sedum plumbizincicola. Shoot biomass showed no significant difference during the repeated phytoextraction experiment on the four test soils and shoot Cd content showed a decreasing trend with the three consecutive crops in soils S1, S2, and S3 but not in soil S4. The Cd removal rates in soils S1, S2, S3, and S4 were 84.5, 81.6, 45.3, and 32.4%, respectively. Rice straw application increased Cd extraction efficiency by 42.6% but the addition of ethylenediaminedisuccinic acid, biochar or nitrogen had no effect on Cd remediation. Shoot Cd content increased significantly (1.57 and 1.71 times, respectively) at low (S₀-1) and high (S₀-2) sulfur addition rates. Soil extractable-Cd in S₀-1 after the experiment showed no significant difference from the control but was 2.43 times higher in S₀-2 than in the control. These results indicate that S. plumbizincicola shows good prospects for the phytoextraction of Cd from highly polluted soils and that the process can be enhanced by adding straw and/or sulfur to the soil.     

Cadmium hyperaccumulation leads to an increase of glutathione rather than phytochelatins in the cadmium hyperaccumulator Sedum alfredii

Sedum alfredii has been reported to be a cadmium (Cd) hyperaccumulator. Phytochelatins (PCs) and other thiol (SH)-containing compounds have been proposed to play an important role in the detoxification and tolerance of some heavy metals, but it is not clear whether PCs are responsible for Cd hyperaccumulation and tolerance in S. alfredii. In this study, two geographically isolated populations of S. alfredii were studied: one population grew on an old Pb/Zn mine site, while the other on a non-mine site. The mine population of this species exhibited a stronger heavy metal tolerance than in the other population. Root-to-shoot transport of Cd was higher in population located at the mine site than at the non-mine site. Considerable amounts of Cd were accumulated in leaves and stems of mine plants, while most Cd was distributed in roots of non-mine plants. Non-protein SH in plant tissues of two populations were further investigated by a HPLC pre-column derivatization system. Upon exposure to Cd, no PCs were detected in all tissues of mine population, while an appreciable amount of glutathione (GSH) was observed in the descending order of stem>root>leaf. The concentrations of GSH consistently increased with the increase of exogenous Cd concentrations and time. On the contrary, Cd exposure strongly induced the production of PCs (mainly PC(2) and PC(3)) and GSH in plant tissues of non-mine population, and the concentrations of GSH showed an initial drop over the duration of 7-d exposure. The present results provided strong evidence that PCs are not involved in Cd transport, hyperaccumulation and tolerance in mine population of S. alfredii.     

Effects of Soil Type and Genotype on Cadmium Accumulation by Rootstalk Crops: Implications for Phytomanagement

The variations of Cd accumulation in three rootstalk crop species (radish, carrot and potato) were investigated by using twelve cultivars grown in acidic Ferralsols and neutral Cambisols under two Cd treatments (0.3 and 0.6 mg kg^?1) in a pot experiment. The result showed that the total Cd uptake was significantly affected by genotype, soil type and interaction between them, suggesting the importance of selecting proper cultivars for phytoextraction in a given soil type. Among the cultivars tested, potato cultivar Luyin No.1 in Ferralsols and radish cultivar Zhedachang in Cambisols exhibited the highest Cd phytoextraction efficiency in aerial parts (4.45% and 0.59%, respectively) under 0.6 mg kg^?1Cd treatment. Furthermore, the Cd concentrations in their edible parts were below the National Food Hygiene Standard of China (0.1 mg kg^?1, fresh weight). Therefore, phytomanagement of slightly Cd-contaminated soils using rootstalk crops for safe food production combined with long-term phytoextraction was feasible, and potato cultivar Luyin No.1 for Ferralsols and radish cultivar Zhedachang for Cambisols were promising candidates for this approach.     

Optimizing phytoremediation of heavy metal-contaminated soil by exploiting plants' stress adaptation

Soil phytoextraction is based on the ability of plants to extract contaminants from the soil. For less bioavailable metals, such as Pb, a chelator is added to the soil to mobilize the metal. The effect can be significant and in certain species, heavy metal accumulation can rapidly increase 10-fold. Accumulation of high levels of toxic metals may result in irreversible damage to the plant. Monitoring and controlling the phytotoxicity caused by EDTA-induced metal accumulation is crucial to optimize the remedial process, i.e. to achieve maximum uptake. We describe an EDTA-application procedure that minimizes phytotoxicity by increasing plant tolerance and allows phytoextraction of elevated levels of Pb and Cd. Brassica juncea is tested in soil with typical Pb and Cd concentrations of 500 mg kg-1 and 15 mg kg-1, respectively. Instead of a single dose treatment, the chelator is applied in multiple doses, that is, in several small increments, thus providing time for plants to initiate their adaptation mechanisms and raise their damage threshold. In situ monitoring of plant stress conditions by chlorophyll fluorescence recording allows for the identification of the saturating heavy metal accumulation process and of simultaneous plant deterioration.     

Cadmium uptake and xylem loading are active processes in the hyperaccumulator Sedum alfredii

Sedum alfredii is a well known cadmium (Cd) hyperaccumulator native to China; however, the mechanism behind its hyperaccumulation of Cd is not fully understood. Through several hydroponic experiments, characteristics of Cd uptake and translocation were investigated in the hyperaccumulating ecotype (HE) of S. alfredii in comparison with its non-hyperaccumulating ecotype (NHE). The results showed that at Cd level of 10 microM measured Cd uptake in HE was 3-4 times higher than the implied Cd uptake calculated from transpiration rate. Furthermore, inhibition of transpiration rate in the HE has no essential effect on Cd accumulation in shoots of the plants. Low temperature treatment (4 degrees C) significantly inhibited Cd uptake and reduced upward translocation of Cd to shoots for 9 times in HE plants, whereas no such effect was observed in NHE. Cadmium concentration was 3-4-fold higher in xylem sap of HE, as compared with that in external uptake solution, whereas opposite results were obtained for NHE. Cadmium concentration in xylem sap of HE was significantly reduced by the addition of metabolic inhibitors, carbonyl cyanide m-chlorophenylhydrazone (CCCP) and 2,4-dinitrophenol (DNP), in the uptake solutions, whereas no such effect was noted in NHE. These results suggest that Cd uptake and translocation is an active process in plants of HE S. alfredii, symplastic pathway rather than apoplastic bypass contributes greatly to root uptake, xylem loading and translocation of Cd to the shoots of HE, in comparison with the NHE plants.