Electrokinetic-enhanced phytoremediation of uranium-contaminated soil using sunflower and Indian mustard

Abstract

Electrokinetic-enhanced phytoremediation is an effective technology to decontaminate heavy metal contaminated soil. In this study, we examined the effects of electrokinetic treatments on plant uptake and bioaccumulation of U from soils with various U sources. Redistribution of uranium in soils as affected by planting and electrokinetic treatments was investigated. The soil was spiked with 100?mg kg^–1 UO₂, UO₃, and UO₂(NO₃)₂. After sunflower and Indian mustard grew for 60 days, 1 voltage of direct-current was applied across the soils for 9 days. The results indicated that U uptake in both plants were significantly enhanced by electrokinetic treatments from soil with UO₃ and UO₂(NO₃)₂. U was more accumulated in roots than in shoots. Electrokinetic treatments were effective on lowering soil pH near the anode region. Overall, uranium (U) removal efficiency reached 3.4–4.3% from soils with UO₃ and uranyl with both plants while that from soil with UO₂ was 0.7–0.8%. Electrokinetic remediation treatment significantly enhanced the U removal efficiency (5–6%) from soils with UO₃ and uranyl but it was 0.8–1.3% from soil with UO₂, indicating significant effects of U species and electrokinetic enhancement on U bioaccumulation. This study implies the potential feasibility of electrokinetic-enhanced phytoremediation of U soils with sunflower and Indian mustard.     

EDTA and Pb—EDTA accumulation in Brassica juncea grown in Pb—amended soil

Previous studies have shown that EDTA is necessary to solubilize soil Pb and facilitate its transport from the soil to the above ground plant tissues. These studies have also suggested that Pb is accumulated in the plant tissue with transpiration as the driving force. We conducted further studies to evaluate the relationship between EDTA soil treatment, plant transpiration, and plant accumulation of Pb and EDTA. Indian mustard (Brassica juncea) plants were grown in soils containing Pb at three different concentrations (1.5, 3.0 and 4.8 mmol/kg) for 5 weeks before being treated with EDTA concentrations ranging from 0 to 10 mmol/kg. Plant shoots and xylem sap were collected and analyzed for Pb and EDTA content using ICP and HPLC, respectively. Water loss was measured for 7 days following EDTA application. Transpiration was not affected at <5 mmol/kg EDTA but, at 10 mmol/kg EDTA transpiration decreased by 80%, whereas accumulation of Pb and EDTA increased. In the Sassafras soil, Pb and EDTA accumulation in the plant shoots continued to increase as the applied EDTA concentration increased, except at the highest level (10 mmol/kg). In soil amended with 4.8 mmol/kg Pb and 10 mmol/kg EDTA, the concentrations of EDTA and Pb in shoots decreased and visible signs of phytotoxicity were observed. The results presented herein support recent studies in hydroponic systems showing that EDTA and Pb are taken up by the plant and suggest that Pb is translocated in the plant as the Pb-EDTA complex. The results also show that the maximum Pb accumulation by plants occurs by maximizing the concentration of the Pb-EDTA complex based on the EDTA extractable soil Pb. This revised version was published online in July 2006 with corrections to the Cover Date.     

Exploring the selenium phytoremediation potential of transgenic Indian mustard overexpressing ATP sulfurylase or cystathionine-gamma-synthase

Indian mustard (Brassica juncea) plants overexpressing ATP sulfurylase (APS transgenics) were previously shown to have higher shoot selenium (Se) levels and enhanced Se tolerance compared to wild type when supplied with selenate in a hydroponic system. Other transgenic Indian mustard overexpressing cystathionine-gamma-synthase (CGS) showed a higher Se volatilization rate, lower shoot Se levels, and higher Se tolerance than wild type, also in hydroponic studies. In the present study, these APS and CGS transgenics were evaluated for their capacity to accumulate Se from soil that is naturally rich in Se. Wild-type Indian mustard and the Se hyperaccumulator Stanleya pinnata were included for comparison. After 10 weeks on Se soil, the APS transgenics contained 2.5-fold higher shoot Se levels than wild type Indian mustard, similar to those of S. pinnata. The CGS transgenics contained 40% lower shoot Se levels than wild type. Shoot biomass was comparable for all Indian mustard types and higher than that of S. pinnata. These results obtained with these transgenics on soil are in agreement with those obtained earlier using hydroponics. The significance of these findings is that they are the first report on the performance of transgenic plants on Se in soil and show the potential of genetic engineering for phytoremediation.     

A Possible Role for Rhizobacteria in Water Treatment by Plant Roots

By accumulating Cd from solution, roots of hydroponically grown Indian mustard (Brassica juncea L.), a high biomass crop plant, were able to cause substantial reductions in the concentration of Cd in solution. The removal of Cd from solution was linearly correlated with Cd accumulation by roots. Screening of 300 different rhizobacterial isolates identified several that, when inoculated onto roots of Indian mustard, significantly enhanced the total amount of Cd removed from solution. Further investigations revealed that this enhancement was because of an overall increase in root biomass in the rhizobacterial-treated plants. Rhizobacteria were found to accumulate Cd from solution, and ultrastructural observations suggested that rhizobacteria promote the precipitation of Cd on the root surface. By precipitating Cd at the root surface, rhizobacteria reduce the amount of Cd taken up into roots, thereby protecting the plants, and in particular the roots, from the toxic effects of Cd. This reduced Cd toxicity allows for the increased proliferation of roots observed when plants are inoculated with certain rhizobacteria.     

Phytoremediation of arsenic and lead in contaminated soil using Chinese brake ferns (Pteris vittata) and Indian mustard (Brassica juncea)

Field and greenhouse experiments were performed to assess the performance of phytoremediation of arsenic and lead from contaminated soil at an EPA Superfund site (Barber Orchard). Chinese Brake ferns (Pteris vittata) were used to extract arsenic. On average, fern shoot arsenic concentrations were as high as 20 times the soil arsenic concentrations under field conditions. It was estimated that 8 years would be required to reduce the acid-extractable portion of soil arsenic to safe levels (40 mg/kg). The effect of soil pH on arsenic extraction was also investigated. Results indicate that increasing soil pH may improve arsenic removal. Indian mustard plants (Brassica juncea) were used under greenhouse conditions to phytoextract soil lead. EDTA was applied to soil and was found to improve lead extraction. When the EDTA concentration was 10 mmol EDTA/kg soil in soil containing 338 mg Pb/kg soil, mustard plants extracted approximately 32 mg of lead. In conclusion, phytoremediation would be a suitable alternative to conventional remediation techniques, especially for soils that do not require immediate remediation.     

The role of root damage in the chelate-enhanced accumulation of lead by Indian mustard plants

In the present study, increasing ethylenediaminetetraacetic acid (EDTA) concentration from 0 to 0.5 mmol L(-1) resulted in progressive increases in root elongation and in shoot and root dry matter (DM) of Indian mustard seedlings (Brassica juncea. L.) exposed at 0.5 mmol L(-1) of lead (Pb). The highest concentration of Pb in the shoots of Indian mustard reached 1140 mg kg(-1) dry weight (DW) in the treatment with 0.5 mmol L(-1) of Pb + 0.25 mmol L(-1) of EDTA. A significantly positive correlation was found between the concentrations of Pb and EDTA in the shoots of mustard. Roots were pretreated with an MC (methanol:trichloromethane) solution, 0.1 mol L(-1) of HCl, and 65 degrees C hot water. The plants were then exposed to 0.5 mmol L(-1) of Pb + 3 mmol L(-1) of EDTA in solution for 2 d. The pretreatments with MC, HCl, and hot water all increased the concentration of Pb in shoots by 14-, 7-, and 15-fold, respectively, compared with the shoots that had not been pretreated. Therefore, some physiological damage to roots would be useful to enhance the uptake of metal by plants and to minimize the application of doses of chelates in the practical operation of chelate-assisted phytoremediation.     

Distribution, transformation and bioavailability of trivalent and hexavalent chromium in contaminated soil

The purpose of this study was to investigate solid-phase distribution, transformation, and bioavailability of Cr in Cr(III) and Cr(VI) contaminated soils. The effects of EDTA treatment on solid-phase distribution of Cr in soils were also examined. The results show that Cr in both initially Cr(III)- and Cr(VI)-contaminated soils was mainly present in the organic matter bound fraction. Chromium had similar solid-phase distribution and similar overall binding intensity in both Cr(III)- and Cr(VI)-contaminated soils after a growing season. Transformation between Cr(III) and Cr(VI) took place in both Cr(III)- and Cr(VI)-treated soils. Chromium in the Cr(III)-contaminated soils was mostly present as Cr(III), while Cr in Cr(VI)-treated soils was mainly transformed into Cr(III). About 2% of Cr in native non-treated soils was found as Cr(VI). EDTA treatment increased Cr in soluble and exchangeable fraction in Cr(III)-treated soils. In both Cr(III)- and Cr(VI)-contaminated soils, Cr in oxide bound and organic matter bound     

4种植物对133Cs和88Sr污染土壤的修复研究

以露地盆栽的苏丹草、向日葵、芥菜、萝卜4种植物为对象,研究它们对土壤中不同浓度(0、2.5、5.0、10.0、20.0、40.0mg/kg)133Cs、88Sr的吸收积累状况,并比较它们对133Cs、88Sr污染土壤的修复效率。结果显示:(1)4种植物单株生物量在各浓度处理下均表现为向日葵>萝卜>芥菜>苏丹草,但它们对133Cs的吸收能力为萝卜>苏丹草>向日葵>芥菜,单株133Cs累积量为向日葵>萝卜>苏丹草>芥菜,单株88Sr累积量表现为萝卜、向日葵>苏丹草>芥菜,而且4种植物对88Sr的吸收能力均强于133Cs。(2)萝卜在除10.0mg/kg133Cs外的各处理中富集系数均大于1,对土壤中133Cs的吸收能力较强;苏丹草在除5.0mg/kg133Cs处理外的转运系数均大于1,其余3种植物在各处理中的转运系数均低于1;88Sr在萝卜体内从根系向上转运到地上部分的能力明显高于其它3种植物,芥菜、向日葵次之。(3)4种植物对88Sr在体内向上的迁移转运能力均大于133Cs。研究表明,向日葵单株对133Cs、88Sr污染土壤的修复效率最高,萝卜次之,且向日葵和萝卜分别因其生物量和吸收能力优势而对被污染土壤中的133Cs和88Sr具有更强的提取能力。     

Overexpression of enzymes involved in glutathione synthesis enhances tolerance to organic pollutants in Brassica juncea

Transgenic Indian mustard (Brassica juncea) overexpressing y-glutamylcysteine synthetase (ECS) or glutathione synthetase (GS) were shown previously to have two-fold higher levels of glutathione and total nonprotein thiols, as well as enhanced cadmium tolerance and accumulation. Here, the hypothesis was tested that these transgenics have enhanced tolerance to organic pollutants, based on the reasoning that many organic xenobiotics are detoxified via conjugation to glutathione. Both the ECS and GS transgenics showed enhanced tolerance to atrazine: while root growth of wildtype seedlings was inhibited 50% by 100 mg L(-1) atrazine, ECS and GS root growth was inhibited 20-30% (P < 0.05). The tolerance of the transgenics to CDNB (1-chloro-2,4-dinitrobenzene). metolachlor, and phenanthrene was also somewhat higher than wild type, but these differences were not as pronounced. Each of the organics treatments significantly enhanced total nonprotein thiol levels in all plant types (2 to 12-fold). Overall, these results suggest that GSH biosynthesis is limiting for atrazine detoxification in Indian mustard and that overexpression of enzymes involved in GSH biosynthesis offers a promising approach to create plants with the enhanced capacity to tolerate not only heavy metals, but also certain organics.     

Hyperaccumulation of Lead by Roots, Hypocotyls, and Shoots of Brassica juncea

The effects of different concentrations of lead nitrate (10^–5 to 10^–3 M) on root, hypocotyl, and shoot growth of Indian mustard (Brassica juncea L. var. megarrhiza), and the uptake and accumulation of Pb²⁺ by its roots, hypocotyls, and shoots were investigated. Lead had no significant inhibitory effect on the root growth at concentrations of 10^–5 to 10^–4 M during the entire treatment, while at 10^–3 M, Pb slightly inhibited the root and shoot growth. B. juncea has ability to take up Pb from solutions and accumulate it in its roots, and transport and concentrate it. The Pb contents in the parts of plants treated with 10^–3 M Pb were greater than those of untreated plants, by factors of 230 in the roots, 170 in the hypocotyls, and 3 in the shoots.     

Chemically enhanced phytoextraction of lead-contaminated soils

The effects of the combined application of soil fungicide (benomyl) and ethylenediaminetetraacetic acid (EDTA) on lead (Pb) phytoextraction by ryegrass (Lolium perenne) were examined. Twenty-five pots of Pb-contaminated soil (200 mg Pb kg(-1)) were seeded with ryegrass and randomly arranged into the following treatments: (1) Control, (2) benomyl, (3) EDTA, (4) benomyl and EDTA (B+E), and (5) benomyl followed by an application of EDTA 14 days later (B .. . E). Chemicals were applied when plants had reached maximum growth. Plants were analyzed for foliage Pb concentration using inductively coupled argon plasma (ICAP) spectrometry. The synergistic effects of the combined benomyl and EDTA application (treatments 4 and 5) were made evident by the significantly (p < 0.05) highest foliage Pb concentrations. However, the foliage dry biomass was significantly lowest for plants in treatments 4 and 5. The bioaccumulation factor (BF) and phytoextraction ratio (PR) were highest for plants in treatment 5 followed by plants in treatment 4.     

向日葵对苯胺废水的光合生理响应及净化效果

为探讨向日葵对苯胺废水的耐受性及其应用于苯胺污染废水植物修复的可行性,采用水培试验方法,以美国油葵1号为材料,研究了不同苯胺浓度胁迫对向日葵光合和叶绿素荧光参数的影响,以及向日葵对苯胺的吸收、积累和净化效果。研究发现:100 mg/L的苯胺对向日葵的净光合速率(P_n)、气孔导度(L_s)和蒸腾速率(T_r)有显著的促进作用,而对生长与叶绿素荧光相关参数无显著的影响。当苯胺浓度升高到200—400 mg/L时,向日葵的鲜重、净光合速率(P_n)、实际光化学效率(Φ_(PSⅡ)),最大光化学效率(F_v/F_m)以及光化学淬灭系数(qP)比对照组显著降低,而胞间CO_2浓度(C_i)显著升高,NPQ呈现出先升高后降低的趋势,并在苯胺浓度为200 mg/L时达到最大值。综合分析表明,当苯胺浓度升高到200 mg/L到400 mg/L时,苯胺对向日葵的净光合速率的抑制以非气孔因素为主。当苯胺浓度为500 mg/L时,导致向日葵死亡。另外,向日葵对100 mg/L苯胺废水中苯胺的去除率最高,达到80.97%。苯胺主要在向日葵的地上部分积累,随着苯胺浓度的升高,向日葵中叶片苯胺的浓度逐渐升高,茎中的苯胺含量变化不显著,而根中的苯胺含量较低。     

Enhancement of cr(lll) phytoaccumulation

Chromium (III) accumulation in high biomass agricultural crops, sunflower (Helianthus annum) and Indian mustard (Brassica juncea) was studied using four soils (pH 4.6 to 7.6) contaminated with different rates of CrCl₃.6H₂O in the presence of synthetic chelate and organic acids. Chromium is essential for normal glucose metabolism in humans and animals, but its contamination and recovery from soils is of environmental concern. Adding ethylenediaminetetraacetic acid (EDTA), citric acid, or oxalic acid to Cr(III)‐contaminated soils significantly increased Cr concentration in plant shoots and roots. Adding Cr(III) complexes of EDTA, citric acid, and oxalic acid to soils dramatically increased (>200‐fold) Cr concentration in shoots and roots. Plant growth was severely decreased but was dependent on soil type, chelate rate, form, and time of chelate application. Chelates and organic acids enhanced Cr(III) accumulation, but its toxic effects were not avoided. Chromium(III) complexes were as toxic to plants as Cr(VI). The phytoaccumulation and recovery of Cr(III) from soils were limited and depended on soil type.     

铬渣堆放场地土壤的污染过程、影响因素及植物修复

铬渣堆放场地的土壤污染已成为重要的环境问题之一,并引起广泛关注。为了对铬渣堆场铬污染情况有更加详细的了解,本文对铬渣污染土壤的2个基本过程(铬渣中铬的水平迁移过程与垂直运移过程)、土壤有机质、pH、Eh和含水量、土壤类型及其无机胶体组成以及地下水运动方向等影响其迁移的因素进行了分析。在此基础上,对铬超积累植物的筛选、铬超积累植物的富集机制、铬渣堆场周围污染土壤的植物修复及其机理进行了概述。虽然目前对铬污染土壤的植物修复还处于起步阶段,但利用超积累植物对铬渣污染场地进行修复前景广阔。     

Insect Diversity in Phytoremediation and Bioaccumulation of Se

A variety of plant species are being considered for the phytoremediation of selenium (Se) contaminated soils in agricultural regions of central California. Use of this plant-based technology may also attract a wide range of insects to these Se-accumulating plants. The first field study surveyed the diversity of insects attracted to tall fescue, birdsfoot trefoil, kenaf, and Indian mustard. Over 7500 specimens were collected by a sweep net collection technique for one complete growing season. Most of the 84 families identified were associated with beneficial insects, although pestiferous insects, for example, thrips, aphids, lygus, and leafhoppers, were also found. In the second study the bioaccumulation of Se in the cabbage looper [Trichoplusia ni (Hübner)] was investigated on Indian mustard grown in Se-rich water culture solution. Neonate larvae were transferred to plants and fed on Se-treated and no Se treated plants (controls) for 14 days, respectively. Pupae were collected from each treatment and incubated until adult insects emerged. Almost 50% fewer pupae were collected from Se-treated plants compared with “controls”, resulting in fewer adult insects. Selenium concentrations were as high as 3173 μg Se kg^-1 DW in adult insects hatched from Se-treated plants compared with <5 μg Se kg^-1 DW in insects from “controls”. Based on both studies, we concluded that insect diversity should be determined and insects monitored for bioaccumulation of Se on phytoremediation sites in agricultural regions.     

Chelate-Enhanced Phytoremediation of Soils Polluted with Heavy Metals

In general, hyperaccumulators are low biomass, slow-growing plants. High biomass non-hyperaccumulator plants by themselves are not a valid alternative for phytoextraction as they also have many limitations, such as small root uptake and little root-to-shoot translocation. In this context, chemically-induced phytoextraction (based on the fact that the application of certain chemicals, mostly chelating agents, to the soil significantly enhances metal accumulation by plants) has been proposed as an alternative for the cleaning up of metal polluted soils. But chelate-induced phytoextraction increases the risk of adverse environmental effects due to metal mobilization during extended periods of time. In order to minimize the phytotoxicity and environmental problems associated with the use of chelating agents, nowadays, research is being carried out on the gradual application of small doses of the chelating agent during the growth period. However, EDTA utilization in the future will most likely be limited to ex situconditions where control of the leachates can be achieved. There are other mobilizing agents which are much less harmful to the environment such as citric acid, NTA, and particularly EDDS. Research should also be aimed towards more innovative agronomic practices. Environmentally safe methods of chelate-induced phytoextraction must be developed before steps towards further development and commercialization of this remediation technology are taken. Most importantly, more applied projects in this field are needed to clarify the real potential and risks of this technology.     

Genetic Variability in the Progeny of Androgenic Dihaploid Plants and Selection of High Agronomic Performing Lines in Brassica Juncea

Androgenic lines of Brassica juncea cv. PR-45 raised by anther culture, were screen for genetic variation. 393 androgenic plants were transferred to pots to study the R₀ generation. These plants showed substantial variation for different characters. Seed progenies of 27 lines of R₀ plants were sown in the field to study R₁ generation. Androgenic plants within lines were significantly homogeneous for the various agronomic characters studied. Two lines were shorter (18 – 20 %) than the control plants, with a remarkable feature of early maturation. Three lines showed 27 – 31 % higher yield than the parent cultivar.