The hyperaccumulator Sedum plumbizincicola harbors metal-resistant endophytic bacteria that improve its phytoextraction capacity in multi-metal contaminated soil

Endophyte-assisted phytoremediation has recently been suggested as a successful approach for ecological restoration of metal contaminated soils, however little information is available on the influence of endophytic bacteria on the phytoextraction capacity of metal hyperaccumulating plants in multi-metal polluted soils. The aims of our study were to isolate and characterize metal-resistant and 1-aminocyclopropane-1-carboxylate (ACC) utilizing endophytic bacteria from tissues of the newly discovered Zn/Cd hyperaccumulator Sedum plumbizincicola and to examine if these endophytic bacterial strains could improve the efficiency of phytoextraction of multi-metal contaminated soils. Among a collection of 42 metal resistant bacterial strains isolated from the tissues of S. plumbizincicola grown on Pb/Zn mine tailings, five plant growth promoting endophytic bacterial strains (PGPE) were selected due to their ability to promote plant growth and to utilize ACC as the sole nitrogen source. The five isolates were identified as Bacillus pumilus E2S2, Bacillus sp. E1S2, Bacillus sp. E4S1, Achromobacter sp. E4L5 and Stenotrophomonas sp. E1L and subsequent testing revealed that they all exhibited traits associated with plant growth promotion, such as production of indole-3-acetic acid and siderophores and solubilization of phosphorus. These five strains showed high resistance to heavy metals (Cd, Zn and Pb) and various antibiotics. Further, inoculation of these ACC utilizing strains significantly increased the concentrations of water extractable Cd and Zn in soil. Moreover, a pot experiment was conducted to elucidate the effects of inoculating metal-resistant ACC utilizing strains on the growth of S. plumbizincicola and its uptake of Cd, Zn and Pb in multi-metal contaminated soils. Out of the five strains, B. pumilus E2S2 significantly increased root (146%) and shoot (17%) length, fresh (37%) and dry biomass (32%) of S. plumbizincicola as well as plant Cd uptake (43%), whereas Bacillus sp. E1S2 significantly enhanced the accumulation of Zn (18%) in plants compared with non-inoculated controls. The inoculated strains also showed high levels of colonization in rhizosphere and plant tissues. Results demonstrate the potential to improve phytoextraction of soils contaminated with multiple heavy metals by inoculating metal hyperaccumulating plants with their own selected functional endophytic bacterial strains.

Variations inter- et intra-spécifiques de la phytoextraction des éléments traces As, Cd, Cu, Ni, Pb et Zn chez trois cultivars de saule

La phytoextraction représente une solution environnementale prometteuse face au problème de contamination des sols en éléments traces (É.T). La présente étude s’intéresse aux différences intra et interspécifiques (S. purpurea, S. dasyclados, S. miyabeana) de trois cultivars de saule lorsqu’ils sont utilisés pour la phytoextration de six É.T. (As, Cd, Cu, Ni, Pb et Zn). Les objectifs sont (i) décrire les variations intrapécifiques du cultivar FISH CREEK (S. purpurea) lorsqu’il est utilisé pour la phytoextraction sur deux sites d’étude; et (ii) décrire les variations intra et interspécifiques des cultivars FISH CREEK (S. purpurea), SV1 (S. dasyclados) et SX67 (S. miyabeana) lorsqu’ils sont utilisés pour la phytoextraction d’un site d’étude. Les indicateurs de variations intra et interspécifiques sélectionnés sont les suivants : la biomasse totale, les concentrations en É.T. extraits et les facteurs de translocation (x ̅ pondérée des conc. É.T. dans les parties aériennes / conc. É.T. dans les racines). La contribution des propriétés du sol (degré de contamination, caractéristiques physicochimiques) à la phytoextraction a été évaluée. Les cultivars ont présenté des variations interspécifiques significatives. Cependant, les variations intraspécifiques sur un site d’étude étaient parfois plus importantes que celles mesurées entre les trois différents cultivars. L’amplitude des variations intraspécifiques que présentent le cultivar FISH CREEK sur deux sites d’étude est attribuée à l’influence du pH, de la minéralogie et au contenu en matière organique, lesquelles diffèrent entre les deux sites. Il a aussi été démontré que la phytoextraction des É.T. n’était pas systématiquement corrélée de façon positive avec le degré de contamination. Cela suggère que les concentrations en É.T. mesurées dans le sol ne peuvent pas expliquer à elles seules la variation des concentrations mesurées dans les tissus. L’implication des mécanismes de rétention dans le sol semblent être davantage responsable des variations observées. La compartimentation des É.T. suggère que le saule est efficace pour l’extraction du Cd et du Zn et qu’il est efficace pour la phytostabilisation de l’As, du Cu, du Ni, et du Pb. En ce qui concerne les quantités extraites, le cultivar FISH CREEK semble le plus performant dans la présente étude.

Enhanced phytoextraction: II. Effect of EDTA and citric acid on heavy metal uptake by Helianthus annuus from a calcareous soil

High biomass producing plant species, such as Helianthus annuus, have potential for removing large amounts of trace metals by harvesting the aboveground biomass if sufficient metal concentrations in their biomass can be achieved. However, the low bioavailability of heavy metals in soils and the limited translocation of heavy metals to the shoots by mosthigh biomass producing plant species limit the efficiency of the phytoextraction process. Amendment of a contaminated soil with ethylene diamine letraacetic acid (EDTA) or citric acid increases soluble heavy metal concentrations, potentially rendering them more available for plant uptake. This article discusses the effects of EDTA and citric acid on the uptake of heavy metals and translocation to aboveground harvestable plant parts in Helianthus annuus. EDTA was included in the research for comparison purposes in our quest for less persistent alternatives, suitable for enhanced phytoextraction. Plants were grown in a calcareous soil moderately contaminated with Cu, Pb, Zn, and Cd and treated with increasing concentrations of EDTA (0. 1, 1, 3, 5, 7, and 10 mmol kg(-1) soil) or citric acid (0. 01, 0. 05, 0.25, 0.442, and 0.5 mol kg(-1) soil). Heavy metal concentrations in harvested shoots increased with EDTA concentration but the actual amount of phytoextracted heavy metals decreased at high EDTA concentrations, due to severe growth depression. Helianthus annuus suffered heavy metal stress due to the significantly increased bioavailable metal fraction in the soil. The rapid mineralization of citric acid and the high buffering capacity of the soil made citric acid inefficient in increasing the phytoextracted amounts of heavy metals. Treatments that did not exceed the buffering capacity of the soil (< 0.442 mol kg(-1) soil) did not result in any significant increase in shoot heavy metal concentrations. Treatments with high concentrations resulted in a dissolution of the carbonates and compaction of the soil. These physicochemical changes caused growth depression of Helianthus annuus. EDTA and citric acid added before sowing of Helianthus annuus did not appear to be efficient amendments when phytoextraction of heavy metals from calcareous soils is considered.

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.

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 soilamendments 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 (similar to soluble fraction), extraction with 1 M NH4OAc at pH 7 (similar to exchangeable fraction), and extraction with 0.5 M NH4OAc + 0.5 M HOAc + 0.02 M EDTA atpH 4.65 (similar to 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 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 annutis will be presented.

Potential of Brassic rapa, Cannabis sativa, Helianthus annuus and Zea mays for phytoextraction of heavy metals from calcareous dredged sediment derived soils

Remediation of soil pollution is one of the many current environmental challenges. Anthropogenic activity has resulted in the contamination of extended areas of land, the remediation of which is both invasive and expensive by conventional means. Phytoextraction of heavy metals from contaminated soils has the prospect of being a more economic in situ alternative. In addition, phytoextraction targets ecotoxicologically the most relevant soil fraction of these metals, i.e. the bioavailable fraction. Greenhouse experiments were carried out to evaluate the potential of four high biomass crop species in their potential for phytoextraction of heavy metals, with or without with the use of soil amendments (EDTA or EDDS). A calcareous dredged sediment derived surface soil, with high organic matter and clay content and moderate levels of heavy metal pollution, was used in the experiments. No growth depression was observed in EDTA or EDDS treated pots in comparison to untreated controls. Metal accumulation was considered to be low for phytoextraction purposes, despite the use of chelating agents. The low observed shoot concentrations of heavy metals were attributed to the low phytoavailability of heavy metals in this particular soil substrate. The mobilising effects induced by EDTA in the soil were found to be too long-lived for application as a soil amendment in phytoextraction. Although EDDS was found to be more biodegradable, higher effect half lives were observed than reported in literature or observed in previous experiments. These findings caution against the use of any amendment, biodegradable or otherwise, without proper investigation of its effects and the longevity thereof. (C) 2005 Elsevier Ltd. All rights reserved.

Comparison of EDTA and EDDS as potential soil amendments for enhanced phytoextraction of heavy metals

Phytoextraction has been proposed as an alternative remediation technology for soils polluted with heavy metals or radionuclides, but is generally conceived as too slow working. Enhancing the accumulation of trace pollutants in harvestable plant tissues is a prerequisite for the technology to be practically applicable. The chelating aminopolycarboxylic acid, ethylene diamine tetraacetate (EDTA), has been found to enhance shoot accumulation of heavy metals. However, the use of EDTA in phytoextraction may not be suitable due to its high environmental persistence, which may lead to groundwater contamination. This paper aims to assess whether ethylene diamine disuccinate (EDDS), a biodegradable chelator, can be used for enhanced phytoextraction purposes. A laboratory experiment was conducted to examine mobilisation of Cd, Cu, Cr, Ni, Pb and Zn into the soil solution upon application of EDTA or EDDS. The longevity of the induced mobilisation was monitored for a period of 40 days after application. Estimated effect half lives ranged between 3.8 and 7.5 days for EDDS, depending on the applied dose. The minimum observed effect half life of EDTA was 36 days, while for the highest applied dose no decrease was observed throughout the 40 day period of the mobilisation experiment. Performance of EDTA and EDDS for phytoextraction was evaluated by application to Helianthus annuus. Two other potential chelators, known for their biodegradability in comparison to EDTA, were tested in the plant experiment: nitrilo acetic acid (NTA) and citric acid. Uptake of heavy metals was higher in EDDS-treated pots than in EDTA-treated pots. The effects were still considered insufficiently high to consider efficient remediation. This may be partly due to the choice of timing for application of the soil amendment. Fixing the time of application at an earlier point before harvest may yield better results. NTA and citric acid induced no significant effects on heavy metal uptake. (C) 2004 Elsevier Ltd. All rights reserved.

Phytoextraction of heavy metals from mine soils using hyperaccumulator plants.

Phytoextraction is an environmental-friendly and cost-effective technology that uses metal hyperaccumulator plants to remove heavy metals from soils. The metals are absorbed by the roots, transported and accumulated in the aerial parts of the plants, which can be harvested and eliminated. The aim of this work was to study some hyperaccumulator species that could be useful to decontaminate mine soils and also to investigate the bioavailability and uptake of these metals by plants with the addition of organic amendments. Pot experiments were performed with soil samples collected from two mining areas in the north of Madrid, where there was an intense mining activity more than 50 years ago. Three species (Thlaspi arvense, Brassica juncea and Atriplex halimus) were grown under controlled conditions in pots filled with contaminated soils mixed with 0 Mg, 30 Mg and 60 Mg per hectare of two different organic amendments: a commercial compost made of pine bark, peat and wood fiber and other made of horse and sheep manure and wood fiber. Plants were harvested at the end of their crop cycle and were digested in order to measure metal concentration (Zn, Cu and Cd) in roots and shoots. Highest plant metal concentration was observed in pots treated with pine bark amendment and with pure soil due to an increase in metal bioavailability with decreasing pH. Also in those treatments the total plant biomass was lower, even some plants could not germinate. On the contrary, there was a lower metal concentration in plant tissues of pots with manure because its higher pH whereas plant growth was significantly larger so there was an incresing amount of metals removed from soil by plants. Comparing the three species results indicate a higher total metal uptake in A. halimus than B. juncea and T. arvense. In conclusion, results show that pH affects metal bioavailability and uptake by hyperaccumulator plants. Addition of organic amendments could be a successful technique for stabilization of metals in contaminated soils.

Phytoextraction of heavy metals from mine soils using hyperaccumulator plants

The aim of this work was to study some hyperaccumulator species that could be useful to decontaminate mine soils and also to investigate the bioavailability and uptake of these metals by plants with the addition of organic amendments. Pot experiments were performed with soil samples collected from two mining areas in the north of Madrid, where there was an intense mining activity more than 50 years ago. Three species (Thlaspi arvense, Brassica juncea and Atriplex halimus) were grown under controlled conditions in pots filled with contaminated soils mixed with 0 Mg, 30 Mg and 60 Mg per hectare of two different organic amendments: a commercial compost made of pine bark, peat and wood fiber and other made of horse and sheep manure and wood fiber. Plants were harvested at the end of their crop cycle and were digested in order to measure metal concentration (Zn, Cu and Cd) in roots and shoots. Highest plant metal concentration was observed in pots treated with pine bark amendment and with pure soil due to an increase in metal bioavailability with decreasing pH. Also in those treatments the total plant biomass was lower, even some plants could not germinate. On the contrary, there was a lower metal concentration in plant tissues of pots with manure because its higher pH whereas plant growth was significantly larger so there was an incresing amount of metals removed from soil by plants. Comparing the three species results indicate a higher total metal uptake in A. halimus than B. juncea and T. arvense. In conclusion, results show that pH affects metal bioavailability and uptake by hyperaccumulator plants. Addition of organic amendments could be a successful technique for stabilization of metals in contaminated soils.

Cadmium phytoextraction capacity of white lupine ( Lupinus albus L.) and narrow-leafed lupine ( Lupinus angustifolius L.) in three contrasting agroclimatic conditions of Chile

The phytoextraction process implies the use of plants to promote the elimination of metal contaminants in the soil. In fact, metal-accumulating plants are planted or transplanted in metal-contaminated soil and cultivated in accordance with established agricultural practices. The objective of the present study was to evaluate the productivity and Cd phytoextraction capacity of white lupine ( Lupinus albus L.) and narrow-leafed lupine ( Lupinus angustifolius L.), as well as the effect on residual Cd concentration in the soil. Both species of lupines were grown at three CdCl2 rates (0, 1, and 2 mg kg-1), under three agroclimatic conditions in Chile in 2013. In the arid zone (Pan de Azúcar, 73 mm precipitation), narrow-leafed lupine production was significantly (P < 0.05) higher than white lupine (4.55 vs. 3.26 Mg DM ha-1, respectively). In locations with higher precipitation (Santa Rosa, 670 mm; Carillanca, 880 mm), narrow-leafed lupine DM production was slightly higher than in Pan de Azúcar, but white lupine was approximately three times higher. Total plant Cd concentrations in white and narrow-leafed lupine increased as Cd rates increased in the three environments, but they were much higher in narrow-leafed lupine than white lupine; 150%, 58%, and 344% higher in Pan de Azúcar, Santa Rosa, and Carillanca, respectively. Cadmium uptake (g Cd ha-1) and apparent recovery were also higher (P < 0.05) in narrow-leafed lupine in two environments (Pan de Azúcar and Carillanca). These results suggest that narrow-leafed lupine present higher potential as phytoremediation species than white lupine.

Bacterial stimulation of copper phytoaccumulation by bioaugmentation with rhizosphere bacteria

Copper contaminated areas pose environmental health risk to living organisms. Remediation processes are thus required for both crop production and industrial activities. This study employed bioaugmentation with copper resistant bacteria to improve phytoremediation of vineyard soils and copper mining waste contaminated with high copper concentrations. Oatmeal plant (Avena sativa L) was used for copper phytoextraction. Three copper resistant bacterial isolates from oatmeal rhizosphere (Pseudomonas putida A1 Stenotrophomonas maltophilia A2 and Acinetobacter calcoaceticus A6) were used for the stimulation of copper phytoextraction. Two long-term copper contaminated vineyard soils (Mollisol and Inceptisol) and copper mining waste from Southern Brazil were evaluated. Oatmeal plants substantially extracted copper from vineyard soils and copper mining waste. As much as 1549 mg of Cu kg(-1) dry mass was extracted from plants grown in Inceptisol soil. The vineyard Mollisol copper uptake (55 mg Cu kg(-1) of dry mass) in the shoots was significantly improved upon inoculation of oatmeal plants with isolate A2 (128 mg of Cu kg(-1) of shoot dry mass). Overall oatmeal plant biomass displayed higher potential of copper phytoextraction with inoculation of rhizosphere bacteria in vineyard soil to the extent that 404 and 327 g ha(-1) of copper removal were respectively observed in vineyard Mollisol bioaugmented with isolate A2 (S. maltophilia) and isolate A6 (A. calcoaceticus). Results suggest potential application of bacterial stimulation of phytoaccumulation of copper for biological removal of copper from contaminated areas. (C) 2010 Elsevier Ltd. All rights reserved.

Behavior of Eucalyptus grandis and E. cloeziana seedlings grown in arsenic-contaminated soil

Arsenic has been considered the most poisonous inorganic soil pollutant to living creatures. For this reason, the interest in phytoremediation species has been increasing in the last years. Particularly for the State of Minas Gerais, where areas of former mining activities are prone to the occurrence of acid drainage, the demand is great for suitable species to be used in the revegetation and "cleaning" of As-polluted areas. This study was carried out to evaluate the potential of seedlings of Eucalyptus grandis (Hill) Maiden and E. cloeziana F. Muell, for phytoremediation of As-polluted soils. Soil samples were incubated for a period of 15 days with different As (Na2HAsO4) doses (0, 50, 100, 200, and 400 mg dm-3). After 30 days of exposure the basal leaves of E. cloeziana plants exhibited purple spots with interveinal chlorosis, followed by necrosis and death of the apical bud at the 400 mg dm-3 dose. Increasing As doses in the soil reduced root and shoot dry matter, plant height and diameter in both species, although the reduction was more pronounced in E. cloeziana plants. In both species, As concentrations were highest in the root system; the highest root concentration was found in E. cloeziana plants (305.7 mg kg-1) resulting from a dose of 400 mg dm-3. The highest As accumulation was observed in E. grandis plants, which was confirmed as a species with potential for As phytoextraction, tending to accumulate As in the root system and stem.

Efficacité d'espèces ligneuses en symbiose mycorhizienne arbusculaire pour la phytoremédiation d'un site urbain contaminé

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Avaliação do potencial fitorremediador da mamona (Ricinus communis L.) e girassol (Helianthus annuus L.) quanto à remoção de chumbo e tolueno em efluentes sintéticos.

The development of research that aim to reduce or even eliminate the environmental impacts provided by anthropogenic actions. One of these main action is the discard of industrial waste in the biotic compartments such as soil, water and air, gained more space in academic settings and in private. A technique of phytoremediation involving the use of plants (trees, shrubs, creepers and aquatic) and their associated microorganisms in order to remove, degrade or isolate toxic substances to the environment. This study aimed to evaluate the potential for phytoremediation of castor bean (Ricinus communis L.) and sunflower (Helianthus annuus L.), wild crops suitable region of Rio Grande do Norte, to reduce concentrations of lead and toluene present in synthetic wastewater that simulate the characteristics of treated water production originated in the petrochemical Guamaré. The experiment was accomplished in randomized blocks in four replicates. Seeds of BRS Energy for the development of seedlings of castor beans and sunflower for Catissol 01, both provided by EMPARN (Empresa de Pesquisa Agropecuária do Rio Grande do Norte) were used. Lead concentrations tested were 250, 500 and 1000 mg/L called T2, T3 and T4, respectively, for toluene the concentrations used were 125, 256 and 501 μg/L, called T5, T6 and T7, respectively. The data for removal of lead in relation to sewage systems applied in castor bean and sunflower were 43.89 and 51.85% (T2), 73.60 and 73.74% (T3) and 85.66 and 87.80 % (T4), respectively, and toluene were approximately 52.12 and 25.54% (T5), 55.10 and 58.05% (T6) and 79.77 and 74.76% (T7) for castor and sunflower seeds, respectively. From the data obtained, it can be deduce that mechanisms involved in reducing the contaminants were of phytoextraction, in relation to lead and phytodegradation for toluene. However, it can be concluded that the castor bean and sunflower crops can be used in exhaust after-treatment of industrial effluents that have this type of contaminant

Phytoremediation of mercury by terrestrial plants

Mercury (Hg) pollution is a global environmental problem. Numerous Hg-contaminated sites exist in the world and new techniques for remediation are urgently needed. Phytoremediation, use of plants to remove pollutants from the environment or to render them harmless, is considered as an environment-friendly method to remediate contaminated soil in-situ and has been applied for some other heavy metals. Whether this approach is suitable for remediation of Hg-contaminated soil is, however, an open question. The aim of this thesis was to study the fate of Hg in terrestrial plants (particularly the high biomass producing willow, Salix spp.) and thus to clarify the potential use of plants to remediate Hg-contaminated soils. Plants used for phytoremediation of Hg must tolerate Hg. A large variation (up to 30-fold difference) was detected among the six investigated clones of willow in their sensitivity to Hg as reflected in their empirical toxicity threshold (TT95b), the maximum unit toxicity (UTmax) and EC50 levels. This gives us a possibility to select Hg-tolerant willow clones to successfully grow in Hgcontaminated soils for phytoremediation. Release of Hg into air by plants is a concern when using phytoremediation in practice. No evidence was found in this study that Hg was released to the air via shoots of willow, garden pea (Pisum sativum L. cv Faenomen), spring wheat (Triticum aestivum L. cv Dragon), sugar beet (Beta vulgaris L. cv Monohill), oil-seed rape (Brassica napus L. cv Paroll) and white clover (Trifolium repens L.). Thus, we conclude that the Hg burden to the atmosphere via phytoremediation is not increased. Phytoremediation processes are based on the ability of plant roots to accumulate Hg and to translocate it to the shoots. Willow roots were shown to be able to efficiently accumulate Hg in hydroponics, however, no variation in the ability to accumulate was found among the eight willow clones using CVAAS to analyze Hg content in plants. The majority of the Hg accumulated remained in the roots and only 0.5-0.6% of the Hg accumulation was translocated to the shoots. Similar results were found for the five common cultivated plant species mentioned above. Moreover, the accumulation of Hg in willow was higher when being cultivated in methyl-Hg solution than in inorganic Hg solution, whereas the translocation of Hg to the shoots did not differ. The low bioavailability of Hg in contaminated soil is a restricting factor for the phytoextraction of Hg. A selected tolerant willow clone was used to study whether iodide addition could increase the plant-accumulation of Hg from contaminated soil. Both pot tests and field trials were carried out. Potassium iodide (KI) addition was found to mobilize Hg in contaminated soil and thus increase the bioavailability of Hg in soils. Addition of KI (0.2–1 mM) increased the Hg concentrations up to about 5, 3 and 8 times in the leaves, branches and roots, respectively. However, too high concentrations of KI were toxic to plants. As the majority of the Hg accumulated in the roots, it might be unrealistic to use willow for phytoextraction of Hg in practice, even though iodide could enhance the phytoextraction efficiency. In order to study the effect of willow on various soil fractions of Hg-contaminated soil, a 5-step sequential soil extraction method was used. Both the largest Hg-contaminated fractions, i.e. the Hg bound to residual organic matter (53%) and sulphides (43%), and the residual fraction (2.5%), were found to remain stable during cultivations of willow. The exchangeable Hg (0.1%) and the Hg bound to humic and fulvic acids (1.1%) decreased in the rhizospheric soil, whereas the plant accumulation of Hg increased with the cultivation time. The sum of the decrease of the two Hg fractions in soils was approximately equal to the amount of the Hg accumulated in plants. Consequently, plants may be suitable for phytostabilization of aged Hg-contaminated soil, in which root systems trap the bioavailable Hg and reduce the leakage of Hg from contaminated soils.