Effects of microbial inoculations and surfactant levels on biologically- and chemically-assisted phytoremediation of lead-contaminated soil by maize (Zea Mays L.)

Assisted phytoremediation has been widely used for decontamination of potentially toxic elements contaminated soils. A greenhouse experiment was conducted to evaluate the effectiveness of different microbial inoculations and surfactant levels on the phytoremediation of a Pb-polluted calcareous soil by maize. The results showed that application of surfactant increased both root and shoot dry matter yields. Microbial inoculations, however, had no significant effect on the root or shoot dry matter yield. Mean Pb uptake in maize root or shoot increased only following the application of some surfactant levels. Inoculation with microorganisms significantly increased both mean Pb concentration and uptake in maize root but not in maize shoot. Application of 4?mmol surfactant kg?1 along with inoculation with Priformospora indica was effective in increasing Pb phytostabilisation potential. While the application of 2?mmol surfactant kg^?1 along with inoculation with Pseudomonas fluorescens was effective in increasing Pb phytoextraction potential. The fact that the values of translocation efficiencies were low in all treatments, demonstrated the low capability of maize for translocation of Pb from root to shoot. Inoculation with P. fluorescens was the most effective treatment in increasing metal micronutrient uptake. Microbial inoculation and surfactant levels enhanced Pb phytoremediation mostly through phytostabilisation of this metal by maize.     

缓释微胶囊EDTA强化玉米提取土壤中铅铜的效应研究

将EDTA转化成具有缓释性能的微胶囊EDTA（Cap-EDTA）作为螯合剂,采用大宝山矿区周边重金属复合污染农田土壤进行重金属连续批浸提及盆栽试验,比较研究Cap-EDTA和未微胶囊化EDTA（Ncap-EDTA）对土壤铅铜活化的动态变化及其对玉米吸收提取铅铜的影响。结果表明：Cap-EDTA处理土壤溶液中初始增溶的铅铜质量分数显著低于Ncap-EDTA处理,且使土壤溶液中的铅铜质量分数长时间持续保持在适度范围内。添加Cap-EDTA的处理显著提高玉米吸收提取铅铜的效率,如实施3 mmol·kg-1的Cap-EDTA使玉米地上部Pb的积累高达1.26 mg·pot-1,是对照处理的1.9倍,是等量Ncap-EDTA处理的1.4倍。因此,在进行污染土壤植物修复的EDTA调控时,采用微胶囊化EDTA,能降低因直接实施EDTA而带来的污染地下水风险,且显著提高玉米修复效率。     

根表铁膜对水稻铅吸收转运的影响

通过温室土壤盆栽试验研究不同生育期水稻根表铁膜形成对水稻吸收和转运Pb的影响。结果表明,两种水稻根表铁膜形成量(以DCB-Fe含量计)及铁膜中吸附的Pb量均随着生育期的延长而减少。水稻根表铁膜Fe含量与铁膜吸附的Pb量呈显著的正相关关系(r=0.798,p0.01)。水稻根系和茎叶吸收积累Pb随着不同品种和不同生育期而变化。品种NK57籽粒Pb含量显著高于品种YD6,前者为后者的1.9倍。从富集系数和分配比率来看,Pb主要富集在水稻的铁膜和根系中,而积累在茎叶和籽粒中的比率较少。Pb从水稻根表铁膜、根系和茎叶向籽粒中的转运系数在两个水稻品种间差异均不显著。水稻分蘖期和孕穗期根表铁膜量与根系Pb含量均呈显著的正相关关系(p0.05),但是成熟期水稻根表铁膜量与根系、茎叶和籽粒中Pb含量相关性均不显著,说明根表铁膜形成对水稻分蘖期和孕穗期吸收积累Pb有一定影响,但对水稻成熟期根系、茎叶和籽粒吸收积累Pb影响不大。     

The effect of application time of mobilising agents on growth and phytoextraction of lead by Brassica napus from a calcareous mine soil

Phytoremediation of heavy metal contaminated sites is often limited by the low-bioavailability of the contaminants. Complexing agents can help to improve this technique by enhancing heavy metal solubility. Pot experiments were conducted to determine the best time for the application of ethylene diamine tetraacetic acid (EDTA) and sheep manure extract (SME) for phytoremediation of a contaminated soil by Brassica napus. The plant was grown on a mine calcareous soil treated with increasing concentrations of EDTA or SME 30 and 10?days before sowing (T1 and T2) and 10 and 30?days after sowing (T3 and T4). Soil available Pb and lead concentrations in plant organs of Brassica napus increased with EDTA concentration. The EDTA application before seed germination significantly reduced rapeseed seedling emergence and dry weight. However, the actual amount of phytoextracted Pb by rapeseed did not decrease significantly, due to severe growth depression, at high EDTA concentrations when it had applied at a suitable time (T4). SME application after sowing increased plant dry weight and Pb concentration in the soil solution and enhanced the accumulated metal concentrations in shoots and roots. Comparing the effect of EDTA and SME on Pb phytoextraction, the study showed that SME is not more effective than EDTA. The efficiency of EDTA on Pb phytoextraction by Brassica napus depends on the time of application. The most efficient treatment for Pb phytoextraction by rapeseed was application of 2?g EDTA/kg soil at T4.     

EDTA-assisted phytoextraction of lead from lead-contaminated soils by Echinochloa crusgalli var. frumentacea

The herbaceous plant Echinochloa crusgalli var. frumentacea is highly resistant to a wide range of heavy metal concentrations. In this study we tested the phytoextraction capacity of E. crusgalli var. frumentacea. Specifically, we compared the effect of EDTA on lead (Pb) accumulation in two groups of plants: those sown in lead contaminated soil and those transplanted to the contaminated soil as seedlings. The result of the time development of the Pb concentrations in the plants in the seedling and seed groups shows that for the seedling group, the effect of adding EDTA to the Pb-contaminated soil was even more pronounced in the shoots than the roots, which showed Pb concentrations 32-fold higher. Compared to the seedling group, the Pb concentrations in the roots of plants in the seed group were approximately 5 times higher in controls and 2 to 10 times higher in the presence of EDTA. Collectively, these results might be considered that EDTA elevates the bioavailability of Pb in soil and this native species is particularly suited to use in Pb phytoextraction.     

Influence of lead on growth and nutrient accumulation in canola (Brassica napus L.) cultivars

Canola (Brassica napus L.) is commonly used as a hyper-accumulator for phytoextraction of heavy metals from soil and water. Like many other heavy metals, lead (Pb) contaminates soil, water and air and thus it is a great problem. This study was conducted to investigate toxic effects of Pb on growth and nutrient uptake in four canola cultivars. Each of four cultivars of canola (Con-II, Con-III, Legend and Shiralee) was subjected to four levels of Pb (0, 30, 60 and 90 mg Pb kg(-1) of soil) from lead chloride [PbCl2]. Due to Pb toxicity, plant growth was adversely affected and relatively a severe reduction in root biomass (45.7%) was recorded. The Pb accumulation increased both in shoot and root, the highest being in root. The uptake of different nutrients, i.e., N, P, K, Ca, Mg, Zn, Cu and Mn was reduced (38.4, 32.8, 33.1, 49.6, 7.78, 52.0, 42.6 and 45.9%, respectively) in the shoots and that of N, Fe, Zn, and Cu in the roots (48.5, 33.2, 24.3 and 44.8%, respectively) of all canola cultivars. The root K, P, Zn and Mn and shoot P, Mg and Fe contents were less affected, the concentration of Pb, Ca and Mg in roots of all cultivars. Among canola cultivars Con-II and Con-III performed better than Legend and Shiralee in terms of growth (26.03%) and nutrient accumulation. Overall, plant growth and nutrient accumulation in the canola cultivars was hampered due to the presence of Pb.     

Phytoaccumulation prospects of cadmium and zinc by mycorrhizal plant species growing in industrially polluted soils

The natural vegetation growing along a wastewater channel was subjected to analyze the uptake of Cadmium (Cd) and Zinc (Zn) and their subsequent accumulation in aboveground and underground plant parts. Species which were mycorrhizal and growing in soils receiving industrially contaminated wastewater were collected along with their rhizospheric soil samples. The nearby uncontaminated control (reference) area was also subjected to sampling on similar pattern for comparison. Both Cd and Zn concentrations were significantly higher in soils of the study area as compared to the reference site. Five plant species i.e. Desmostachya bipinnata, Dichanthium annulatum, Malvastrum coromandelianum, Saccharum bengalense, and Trifolium alexandrinum were analyzed for metal uptake. The maximum phytoaccumulation of Cd was observed in Desmostachya bipinnata (20.41 μg g⁻¹) and Dichanthium annulatum (15.22 μg g⁻¹) for shoot and root tissues, respectively. However, Malvastrum coromandelianum revealed maximum Zn accumulation for both the shoot and the root tissues (134 and 140 μg g⁻¹, respectively). The examination of cleared and stained roots of the plants from both the areas studied revealed that all of them were colonized to a lesser or a greater degree by arbuscular mycorrhizal (AM) fungi. The Cd hyperaccumulating grasses i.e. Desmostachya bipinnata and Dichanthium annulatum, from study area had smaller root:shoot (R/S) ratio as compared to those growing on reference area indicating a negative pressure of soil metal contamination. The lower R/S ratio in the mycorrhizal roots observed was probably due to increased AM infection and its mediatory role in soil plant transfer of heavy metals. Furthermore, comparatively lower soil pH values in the study areas may have played a key role in making the overall phytoavailability of both the metals. Consequently variations in Cd and Zn tissue concentration among species were observed that also indicate the phytoaccumulation potential of the native species.     

有机酸对根际土壤中铅形态及其生物毒性的影响

采用根袋盆栽试验和连续浸提方法研究外源柠檬酸、EDTA对根际土壤中Pb形态转化及其生物毒性的影响。结果表明，有机酸能明显活化根际土壤中的Pb，高浓度(3mmol／L)比低浓度(0．5mmol／L)的柠檬酸对Pb的活化能力强；EDTA在低浓度(0．5mmol／L)时对Pb的活化能力就很强。柠檬酸、EDTA能增强土壤Pb的毒性，提高小麦根部对Pb的吸收，并促进Pb由根部向地上部转移。     

Introducing a new metal accumulator plant and the evaluation of its ability in removing heavy metals

A field study was conducted in a dried waste pool of a lead (Pb) mine in Arak (Iran) to find the accumulator plant(s) and to evaluate the amount of metal bioaccumulation in the root and shoot portion of the naturally growing vegetation. Concentrations of heavy metals were determined both in the soil and the plants that were grown in the dried waste pool. The concentrations of total Cu, Zn, Pb, and Ni in the waste pool were found to be higher than the natural soil and the toxic levels. The results showed that six dominant vegetations, namely, Centaurea virgata, Eleagnum angustifolia, Euphorbia macroclada, Gundelia tournefortii, Reseda lutea, and Scariola orientalis accumulated heavy metals. Based on the results, it was concluded that E. macroclada belonging to Euphorbiaceae is the best Pb accumulator and also a good accumulator for Zn, Cu, and Ni. The bioaccumulation ability of E. macroclada was evaluated in experimental pots. The study showed that the amount of heavy metals in polluted soils decreased several times during two years of phytoremediation. The accumulation of metal in the root, leaves, and shoot portions of E. macroclada varied significantly, but all the concentrations were within the toxic limits. Based on the obtained data, E. macroclada is an effective accumulator plant for soil detoxification and phytoremediation in critical conditions.     

Enhanced Ni phytoextraction by effectiveness of chemical and biological amendments in sunflower plant grown in Ni-polluted soils

A greenhouse study was conducted as a completely randomised design in a factorial arrangement to assess how inoculation of AMF (arbuscular mycorrhizal fungus) and application of EDTA (ethylenediaminetetracetic acid) as biological and chemical amendments can affect the Ni (nickel) phytoremediation in Ni-polluted soils using sunflower plant. The results showed that the inoculation of AMF increased root colonisation while applying EDTA and high level of Ni decreased it. Microbial incubation has a positive effect on both shoot and root dry yields; however, co-application of Ni and EDTA demoted the growth rate. Shoot nutrients uptake of plants decreased as Ni levels increased. In inoculated plants, shoot uptake of Zn, Fe and Mn was higher in all Ni levels than non-inoculated plants. Ni uptake in plant shoots and roots increased with applying both AMF and EDTA. However, the mean Ni concentration and uptake in inoculated plants along with applying EDTA are higher in sunflower shoots than in roots. As Ni levels increased, Ni extraction and uptake efficiencies increased; it can be concluded co-application of EDTA and AMF was effective in increasing phytoextraction potential of sunflower plants in Ni-contaminated sites. This study highlights that AMF could be suitable for cleaning Ni-polluted areas and it could significantly contribute to phytoremediation technology.     

Characterisation of growth and biochemical response of Onopordum acanthium L. under lead stress as affected by microbial inoculation

Microbial associations may influence the negative effects of potentially toxic elements on plants. In a greenhouse experiment, the growth; biochemical response; and Pb, Fe, and Zn uptake of Onopordum acanthium L. were investigated in response to inoculation with arbuscular mycorrhizal fungi, AMF (a mixture of Funneliformis mosseae, Rhizophagus irregularis, and Rhizophagus fasciculatus) and plant growth-promoting rhizobacteria, PGPR (a mixture of Pseudomonas species including P. putida, P. fluorescens, and P. aeruginosa) at increased Pb levels in soil. The treatments were arranged as a factorial experiment based on a randomised complete block design. Results revealed that inoculation with AMF and PGPR decreased Pb toxicity in plants. Inoculated plants with AMF and PGPR had higher shoot and root dry weight compared with the non-inoculated plants. In this study, AMF and PGPR inoculation led to a significant increase (P?≤?.05) in chlorophyll a, b, chlorophyll a+b, carotenoid, proline, and relative water content of plants. Furthermore, AMF and PGPR inoculation likely played a more important role in growth and Pb uptake in O. acanthium L. Our results suggest that AMF and Pseudomonas bacteria could be effective bio-inoculants for enhancing the plant growth and Pb uptake by inhibiting the adverse effects of Pb in O. acanthium.     

Arbuscular mycorrhiza confers lead tolerance and uptake in Miscanthus sacchariflorus

The effects of an arbuscular mycorrhizal fungi (AMF) association on the growth, survival capabilities, nutrients and lead (Pb) uptake of Miscanthus sacchariflorus under different Pb concentrations were studied in the form of pot cultures. The treatments comprised inoculation or non-inoculation of the AMF, Gigaspora margarita, and the addition of three Pb concentrations to the soil (0, 100 and 1000?mg?kg^?1). The addition of Pb significantly decreased mycorrhizal colonisation. The inoculation of AMF with Pb increased chlorophyll content, Fv/Fm, total dry mass, indole-3-acetic acid (IAA), total nitrogen, and total phosphorus, whereas H₂O₂ level, indole-3-acetic acid oxidase (IAAO) activity, and peroxidase (POD) activity were low compared to those in the non-inoculated treatments. Moreover, the application of AMF together with Pb doses induces concentrations of Pb in the plant, where the higher dose of Pb (1000?mg?kg^?1) induces a lower content of Pb in the aerial part of the plant but a higher content in the root. G. margarita enhanced the tolerance of M. sacchariflorus against Pb toxicity, and facilitated the accumulation of Pb in the plant roots, whereas translocation to the shoots was inhibited at the highest dose Pb (1000?mg?kg^?1). However, in contaminated soil, the Pb removal capability of M. sacchariflorus with AMF was remarkable.     

Mobilization of heavy metals from contaminated paddy soil by EDDS,EDTA, and elemental sulfur

For enhanced phytoextraction, mobilization of heavy metals (HMs) from the soil solid phase to soil pore water is an important process. A pot incubation experiment mimicking field conditions was conducted to investigate the performance of three soil additives in mobilizing HMs from contaminated paddy soil (Gleyi-Stagnic Anthrosol): the [S, S]-isomer of ethylenediamine disuccinate (EDDS) with application rates of 2.3, 4.3, and 11.8 mmol kg⁻¹ of soil, ethylenediamine tetraacetate (EDTA; 1.4, 3.8, and 7.5 mmol kg⁻¹), and elemental sulfur (100, 200, and 400 mmol kg⁻¹). Temporal changes in soil pore water HM and dissolved organic carbon concentrations and pH were monitored for a period of 119 days. EDDS was the most effective additive in mobilizing soil Cu. However, EDDS was only effective during the first 24 to 52 days, and was readily biodegraded with a half-life of 4.1 to 8.7 days. The effectiveness of EDDS decreased at the highest application rate, most probably as a result of depletion of the readily desorbable Cu pool in soil. EDTA increased the concentrations of Cu, Pb, Zn, and Cd in the soil pore water, and remained effective during the whole incubation period due to its persistence. The highest rate of sulfur application led to a decrease in pH to around 4. This increased the pore water HM concentrations, especially those of Zn and Cd. Concentrations of HMs in the soil pore water can be regulated to a large extent by choosing the proper application rate of EDDS, EDTA, or sulfur. Hence, a preliminary work such as our pot experiment in combination with further plant experiments (not included in this study) will provide a good tool to evaluate the applicability of different soil additives for enhanced phytoextraction of a specific soil.     

Response of roselle (Hibiscus sabdariffa) to heavy metals contamination in soils with different organic fertilisations

The response of green roselle (Hibiscus sabdariffa) to Cu/Pb contamination and manure application in soil was investigated using pot experiments. Subsamples of a mineral soil were treated with increasing doses (0–500 mg kg^?1) of Cu/Pb only and/or amended (at 10% w/w) with poultry or swine manure. Roselle plants were grown, monitored for changes in growth rate and post-harvest aboveground dry biomass and tissue Cu/Pb concentrations were determined. The plants were typically greenish with linear growth profiles at all metal doses, indicating some level of tolerance. Dry biomass yields decreased as metal dose increased. Poultry manure enhanced roselle biomass yields better than swine manure. Tissue Cu/Pb concentrations increased linearly as metal doses increased in unamended soils; whereas nonlinear responses were observed in manure-amended soils. Soil-to-plant transfer factors, T _f (%) indicated that Cu (13≤T _f (% )≤60) was more phytoavailable to roselle than Pb (11≤T _f (% )≤20). Tissue metal concentrations were modelled from soil pH, organic matter, plant available and pseudototal metal; but the models appeared more reliable with plant available metal as a covariate than with pseudototal metal content. These observations may become useful whenever phytoextraction is the remedial option for soils moderately contaminated by toxic metals.     

Trace metal contents ofTaraxacum officinale (dandelion) as a convenient environmental indicator

Some vascular plants are known to concentrate trace metals and are regarded to be suitable indicators of atmospheric metal deposition. Among plant species used for biogeochemical studies dandelion (Taraxacum officinale.) is convenient for monitoring air/soil pollution. The plant commonly occurs in different ecosystems with relatively parallel stages of ontogenesis over a broad area of geographical regions. Its leaves and roots are easily accessible for sampling. Leaf to root ratios of metal concentrations in dandelion may indicate the source of metals during the growing season. Trace metals in leaves and roots of dandelion from 132 sites in Poland showed higher concentrations in the plants from the SW region compared to those from the NE region of the country. However, the differences were only statistically significant ( = 0.05) for Cd, Ni, and Pb. Geometric means of metal concentrations (mg kg^–1, air dried weight) in dandelion leaves of the SW and NE regions were: Cd 0.85, 0.52; Cr 0.99, 0.81; Cu 11.2, 11.1; Fe 184.4, 100.0; Mn 59.7, 51.4; Ni 2.1, 1.5; Pb 4.4, 3.0; and Zn 49.6, 41.3, respectively. Markedly higher concentrations of Cd, Pb, and Zn were found in the leaves of the dandelion over roots in the SW region. These metals are the most serious aerial pollutants in that part of the country.     

Fraction distribution and migration of heavy metals in mangrove-sediment system under sulphur and phosphorus amendment

In this study, mangrove seedlings (Kandelia obovata (S. L.)) were cultivated in rhizo-boxes, which contain sediments collected from natural mangrove forest and modified with different rates of sulphate and phosphate. The fraction distributions of Zn, Cd, Cu, Ni and Pb in rhizosphere and non-rhizosphere sediments were studied by using a sequential extraction method. Metal concentrations in plant tissues and iron plaque on root surface were also determined to reveal migration variation of heavy metals in the plant-sediment system. The results showed that the activities of K. obovata roots enhance the reducible metals while reducing acid-extractable and oxidisable metals; sulphur amendment benefits the combination of metal ions with S^2– and therefore reduces the bioavailability of metal pollutants; addition of sulphur also improves the content of iron plaque on the root surface, which plays an important role in metal accumulation by K. obovata root tissue; the addition of sulphur markedly reduces the concentration of Cd in roots, but significantly enhanced the concentrations of Cu, Zn, Ni and Pb in roots. The results indicate that sulphur and phosphorus content in mangrove sediment, and the growth of mangrove plant can significantly influence the migration of heavy metals in the mangrove wetland ecosystem.     

Accumulation and phytoavailability of benzo[a]pyrene in an acid sandy soil

Effects of benzo[a]pyrene (B[a]P) on ryegrass (Lolium perenne L.) growth, plant accumulation and dissipiation of B[a]P in a red sandy soil (Hapli-Udic Argosol) were studied in a pot experiment. The plants were grown for 61 days in soil spiked with B[a]P at 0, 12.5, 25 and 50 mg kg⁻¹. Control pots without plants were also set up. Soil extractable B[a]P, plant shoot and root biomass, and concentrations of B[a]P in plant shoots and roots were determined. Ryegrass biomass was increased by addition of B[a]P and root B[a]P concentrations were significantly correlated with B[a]P application rate, but no such correlation was found for shoot B[a]P concentrations. This indicates that B[a]P enhanced the growth of the ryegrass. The extractable B[a]P concentration in the planted soil was significantly lower than that in the unplanted control soil at the rate of 50 mg B[a]P kg⁻¹. This indicates that ryegrass may help to dissipate B[a]P in soil at concentrations over 50 mg kg⁻¹ soil although the mechanism for this is not understood.     

Response of Lupinus albus to Pb–EDTA indicates relatively high tolerance

White lupin plants were grown in hydroponics with 0, 90 and 180 µmol L^?1 Pb(II) ethylenediaminetetraacetate complex for 30 days. Pb distribution (shoot/root ratio) was 0.34 and 0.46 for both Pb treatments. In the shoots, no decrease in biomass nor in photosynthetic pigment levels and no changes in the concentrations of malondialdehyde and glutathione were detected. In the roots, malondialdehyde increased by 20%, glutathione 2–3.6 times and phytochelatin concentrations 4–5 times. The high tolerance of white lupin makes it a valuable plant for phytoremediation of Pb-contaminated soil.     

Effects of zinc stress on growth and antioxidant enzyme responses of Kandelia obovata seedlings

This study evaluated the tolerance and accumulation potential in the mangrove Kandelia obovata under moderate and high levels of external Zn. A greenhouse experiment was conducted to investigate the effects of a range of external Zn concentrations (0–400 mg L^?1) on the growth of this species by counting the lateral root number and measuring the root length, leaf area, and total dry biomass. We also determined the Zn accumulation in plant tissues. K. obovata survived with external Zn concentrations of up to 400 mg L^?1, although the excess metal resulted in a biomass reduction of 34%. A significant increase in antioxidant enzyme activities occurred in roots of plants under high-level Zn stress, suggesting that K. obovata seedlings could tolerate up to 200 mg L^?1 zinc treatment.