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.

Rhizosphere Bacteria Enhance Selenium Accumulation and Volatilization by Indian Mustard1

Indian mustard (Brassica juncea L.) accumulates high tissue Se concentrations and volatilizes Se in relatively nontoxic forms, such as dimethylselenide. This study showed that the presence of bacteria in the rhizosphere of Indian mustard was necessary to achieve the best rates of plant Se accumulation and volatilization of selenate. Experiments with the antibiotic ampicillin showed that bacteria facilitated 35% of plant Se volatilization and 70% of plant tissue accumulation. These results were confirmed by inoculating axenic plants with rhizosphere bacteria. Compared with axenic controls, plants inoculated with rhizosphere bacteria had 5-fold higher Se concentrations in roots (the site of volatilization) and 4-fold higher rates of Se volatilization. Plants with bacteria contained a heat-labile compound in their root exudate; when this compound was added to the rhizosphere of axenic plants, Se accumulation in plant tissues increased. Plants with bacteria had an increased root surface area compared with axenic plants; the increased area was unlikely to have caused their increased tissue Se accumulation because they did not accumulate more Se when supplied with selenite or selenomethionine. Rhizosphere bacteria also possibly increased plant Se volatilization because they enabled plants to overcome a rate-limiting step in the Se volatilization pathway, i.e. Se accumulation in plant tissues.

Overexpression of ATP Sulfurylase in Indian Mustard Leads to Increased Selenate Uptake, Reduction, and Tolerance1

In earlier studies, the assimilation of selenate by plants appeared to be limited by its reduction, a step that is thought to be mediated by ATP sulfurylase. Here, the Arabidopsis APS1 gene, encoding a plastidic ATP sulfurylase, was constitutively overexpressed in Indian mustard (Brassica juncea). Compared with that in untransformed plants, the ATP sulfurylase activity was 2- to 2.5-fold higher in shoots and roots of transgenic seedlings, and 1.5- to 2-fold higher in shoots but not roots of selenate-supplied mature ATP-sulfurylase-overexpressing (APS) plants. The APS plants showed increased selenate reduction: x-ray absorption spectroscopy showed that root and shoot tissues of mature APS plants contained mostly organic Se (possibly selenomethionine), whereas wild-type plants accumulated selenate. The APS plants were not able to reduce selenate when shoots were removed immediately before selenate was supplied. In addition, Se accumulation in APS plants was 2- to 3-fold higher in shoots and 1.5-fold higher in roots compared with wild-type plants, and Se tolerance was higher in both seedlings and mature APS plants. These studies show that ATP sulfurylase not only mediates selenate reduction in plants, but is also rate limiting for selenate uptake and assimilation.

Overexpression of Glutathione Synthetase in Indian Mustard Enhances Cadmium Accumulation and Tolerance1

An important pathway by which plants detoxify heavy metals is through sequestration with heavy-metal-binding peptides called phytochelatins or their precursor, glutathione. To identify limiting factors for heavy-metal accumulation and tolerance, and to develop transgenic plants with an increased capacity to accumulate and/or tolerate heavy metals, the Escherichia coli gshII gene encoding glutathione synthetase (GS) was overexpressed in the cytosol of Indian mustard (Brassica juncea). The transgenic GS plants accumulated significantly more Cd than the wild type: shoot Cd concentrations were up to 25% higher and total Cd accumulation per shoot was up to 3-fold higher. Moreover, the GS plants showed enhanced tolerance to Cd at both the seedling and mature-plant stages. Cd accumulation and tolerance were correlated with the gshII expression level. Cd-treated GS plants had higher concentrations of glutathione, phytochelatin, thiol, S, and Ca than wild-type plants. We conclude that in the presence of Cd, the GS enzyme is rate limiting for the biosynthesis of glutathione and phytochelatins, and that overexpression of GS offers a promising strategy for the production of plants with superior heavy-metal phytoremediation capacity.

Rate-Limiting Steps in Selenium Assimilation and Volatilization by Indian Mustard1

Se can be accumulated by plants and volatilized to dimethylselenide, providing an attractive technology for Se phytoremediation. To determine the rate-limiting steps in Se volatilization from selenate and selenite, time- and concentration-dependent kinetics of Se accumulation and volatilization were studied in Indian mustard (Brassica juncea). Time-dependent kinetic studies showed that selenate was taken up 2-fold faster than selenite. Selenate was rapidly translocated to the shoot, away from the root, the site of volatilization, whereas only approximately 10% of the selenite was translocated. For both selenate- and selenite-supplied plants, Se accumulation and volatilization increased linearly with external Se concentration up to 20 μm; volatilization rates were also linearly correlated with root Se concentrations. Se-volatilization rates were 2- to 3-fold higher from plants supplied with selenite compared with selenate. Se speciation by x-ray absorption spectroscopy revealed that selenite-supplied plants accumulated organic Se, most likely selenomethionine, whereas selenate-supplied plants accumulated selenate. Our data suggest that Se volatilization from selenate is limited by the rate of selenate reduction, as well as by the availability of Se in roots, as influenced by uptake and translocation. Se volatilization from selenite may be limited by selenite uptake and by the conversion of selenomethionine to dimethylselenide.

The Role of EDTA in Lead Transport and Accumulation by Indian Mustard1

Indian mustard (Brassica juncea) plants exposed to Pb and EDTA in hydroponic solution were able to accumulate up to 55 mmol kg−1 Pb in dry shoot tissue (1.1% [w/w]). This represents a 75-fold concentration of Pb in shoot tissue over that in solution. A threshold concentration of EDTA (0.25 mm) was found to be required to stimulate this dramatic accumulation of both Pb and EDTA in shoots. Below this threshold concentration, EDTA also accumulated in shoots but at a reduced rate. Direct measurement of a complex of Pb and EDTA (Pb-EDTA) in xylem exudate of Indian mustard confirmed that the majority of Pb in these plants is transported in coordination with EDTA. The accumulation of EDTA in shoot tissue was also observed to be directly correlated with the accumulation of Pb. Exposure of Indian mustard to high concentrations of Pb and EDTA caused reductions in both the transpiration rate and the shoot water content. The onset of these symptoms was correlated with the presence of free protonated EDTA (H-EDTA) in the hydroponic solution, suggesting that free H-EDTA is more phytotoxic than Pb-EDTA. These studies clearly demonstrate that coordination of Pb transport by EDTA enhances the mobility within the plants of this otherwise insoluble metal ion, allowing plants to accumulate high concentrations of Pb in shoots. The finding that both H-EDTA and Pb-EDTA are mobile within plants also has important implications for the use of metal chelates in plant nutritional research.

Potencial de fitorremediação de diferentes plantas em solo contaminado por 14C-tebuthiuron

Este trabalho teve como objetivo avaliar a tolerância e a capacidade de fitorremediação de crotalária (Crotalaria spectabilis), sorgo (Sorghum bicolor), nabo forrageiro (Raphanus sativus), amendoim (Arachis hypogaea) e alfafa (Mendicago sativa) a solos contaminados por tebuthiuron. Para determinar a tolerância das plantas ao herbicida estas foram submetidas a 5 diferentes doses (300, 600, 1200, 2400 e 4800 gramas de ingrediente ativo por hectare gi.a. ha-1) e comparadas com uma testemunha que não recebeu aplicação do herbicida. Crotalária, Nabo forrageiro e alfafa se mostraram sensíveis ao herbicida, mesmo na menor dose de aplicação, enquanto sorgo se mostrou tolerante ao herbicida até a dose de 600 gi.a. ha-1 tendo apresentado fitointoxicação de 80% na dose de 1200 gi.a. ha-1 e amendoim foi tolerante até a dose de 4800 gi.a. ha-1 para a qual apresentou apenas 40% de fitointoxicação. Plantas de amendoim e sorgo foram submetidas a dose de contaminação de 600 gi.a. ha-1 aplicada por meio de uma solução de trabalho contendo 17,47kBq (Quilobecquerels) de 14C-tebuthiuron. As duas plantas foram capazes de remediar o solo, no entanto amendoim se mostrou mais eficiente reduzindo a contaminação em 75,8% enquanto sorgo retirou do solo 44,49% do herbicida contaminante