Cu-contaminated Soil Research Articles

Long-term phytomanagement with compost and a sunflower – Tobacco rotation influences the structural microbial diversity of a Cu-contaminated soil

At a former wood preservation site contaminated with Cu, various phytomanagement options have been assessed in the last decade through physicochemical, ecotoxicological and biological assays. In a field trial at this site, phytomanagement with a crop rotation based on tobacco and sunflower, combined with the incorporation of compost and dolomitic limestone, has proved to be efficient in Cu-associated risk mitigation, ecological soil functions recovery and net gain of economic and social benefits. To demonstrate the long-term effectiveness and sustainability of phytomanagement, we assessed here the influence of this remediation option on the diversity, composition and structure of microbial communities over time, through a metabarcoding approach. After 9 years of phytomanagement, no overall effect was identified on microbial diversity; the soil amendments, notably the repeated compost application, led to shifts in soil microbial populations. This phytomanagement option induced changes in the composition of soil microbial communities, promoting the growth of microbial groups belonging to Alphaproteobacteria, many being involved in N cycling. Populations of Nitrososphaeria, which are crucial in nitrification, as well as taxa from phyla Planctomycetacia, Chloroflexi and Gemmatimonadetes, which are tolerant to metal contamination and adapted to oligotrophic soil conditions, decreased in amended phytomanaged plots. Our study provides an insight into population dynamics within soil microbial communities under long-term phytomanagement, in line with the assessment of soil ecological functions and their recovery.

The effect of Cu-resistant plant growth-promoting rhizobacteria and EDTA on phytoremediation efficiency of plants in a Cu-contaminated soil.

Remediation of heavy metal-contaminated soils is essential for safe agricultural or urban land use, and phytoremediation is among the most effective methods. The success of phytoremediation relies on the size of the plant biomass and bioavailability of the metal for plant uptake. This research was carried out to determine the effect of Ethylenediaminetetraacetic acid (EDTA) ligand and Cu-resistant plant growth-promoting rhizobacteria (PGPR) on phytoremediation efficiency of selected plants as well as fractionation and bioavailability of copper (Cu) in a contaminated soil. The test conditions included three plant species (maize: Zea mays L., sunflower: Helianthus annuus L., and pumpkin: Cucurbita pepo L.) and six treatments, comprising two PGPR strains (Pseudomonas cedrina K4 and Stenotrophomonas sp. A22), two PGPR strains with EDTA, EDTA, and control (without PGPR and EDTA). The combination of EDTA and PGPR enhanced the Cu concentration in both shoot and root tissues and increased the plant biomass. The Cu specific uptake was at a maximum level in the shoots of pumpkin plants when treated with the PGPR strain K4 + EDTA (202μgpot-1), and the minimum amount of Cu was recorded for sunflower with no PGPR or EDTA addition (29.6μgpot-1). The result of the PGPR-EDTA treatments showed that the combined application of EDTA and PGPR increased the shoot Cu-specific uptake approximately fourfold in pumpkin. Pumpkin with the highest shoot Cu specific uptake and maize with the highest root Cu specific uptake were the most effective plants in phytoextraction and phytostabilization, respectively. The effectiveness of different PGPR-EDTA treatments in increasing Cu specific uptake by crop plants was assessed by measuring the amount of Cu extracted from the rhizosphere soil adhering to the roots of crop species, by the use of the single extractants Diethylenetriamine pentaacetic acid (DTPA), H2O, NH4NO3, and NH4OAc. PGPR-EDTA treatments increased the amount of water-extractable Cu from rhizosphere soils more than ten times that of the control. The combined application of the EDTA and PGPR reduced the carbonated Fe and Mn oxide-bound Cu in the contaminated soil, and increased the soluble and exchangeable concentration of Cu. Pumpkin, with high shoot biomass and the highest shoot Cu specific uptake was found to be the most effective field crop in phytoextraction of Cu from the contaminated soil. The results of this pot study demonstrated that the EDTA+PGPR treatment could play an important role in increasing the Cu bioavailability and specific uptake by plants, and thus increasing the phytoremediation efficiency of plants in Cu-contaminated areas.

Biochar increases plant water use efficiency and biomass production while reducing Cu concentration in Brassica juncea L. in a Cu-contaminated soil

Biochar has been recently used as an alternative strategy to improve soil quality and plant growth in metal contaminated soils. However, the effects of biochar on gas exchange parameters such photosynthetic rate (A), water use efficiency (WUE) and instantaneous carboxylation efficiency (ICE) in metal tolerant plant species in contaminated soils is still unknown. Such information is important to understand how different biochar types can influence plant biomass production and metal uptake. Hence, a greenhouse experiment was set up as a completely randomized design combining two types of crop residue biochar (coconut husk (CB) and orange shell (OB) and two rates of application (30 t ha−1 and 60 t ha−1)). A control treatment (no biochar) was also included. The aim of the study was to investigate the influence of biochar on the physiological performance, growth and concentration of copper (Cu) in the shoot of B. juncea plants in a Cu-contaminated soil. Besides reduced growth on the control soil, all other treatments increased plant growth. No toxicity symptom was observed in the plant, confirming its Cu tolerance. Biochar increased plant biomass by approximately 170% and reduced Cu concentration up to 51%. Application of 30 t ha−1 biochar significantly increased net photosynthesis in 59% (CB) and in 34% (OB) while reducing stomatal conductance in 40% and transpiration rate in 14% (CB) and 19% (OB). Application of 60 t ha−1 of biochar reduced stomatal conductance and transpiration rate in 51% and 60% (CB) and 26% and 7% (OB), respectively. All biochars improved the water use efficiency (WUE) in the plants (52–182%) for the instantaneous WUE and 80–162% for the intrinsic (IWUE). Therefore, biochar, especially CB, caused an improved regulation of the stomata aperture so that plant could maintain a high photosynthetic rate while efficiently controlling the use of water. This could be an important mechanism to reduce the excessive uptake of Cu by the plant.

The potential of two different Avena sativa L. cultivars to alleviate Cu toxicity

Agronomic strategies as intercropping might be applied to reduce plant-available copper (Cu) in Cu-contaminated soils. Thus, our aim was to characterize two different oat cultivars, Avena sativa L. cv. Fronteira and cv. Perona for their ability to tolerate and/or phytostabilize Cu.Copper toxicity reduced plant biomass of both cultivars. The exudate analysis revealed the presence of phenolic compounds and phytosiderophores, yet with a different pattern between the cultivars: cv. Fronteira showed a Cu-concentration and time-dependent release of phenolic compounds, while cv. Perona down-regulated this release during the second week of treatment. Copper concentration increased linearly in all the tissues analysed with increasing Cu concentration showing yet a different compartmentalization: cv. Fronteira and cv. Perona preferentially accumulated Cu in the apoplasm and symplast, respectively. This higher accumulation of Cu in the apoplasm strongly reduces the available binding sites, leading to a competitive absorption with other macro-and micronutrients (e.g. Ca, Mn, Zn). Furthermore, in both cultivars Cu toxicity led to a significant reduction of shoot phosphorus content.The ionomic profile and compartmentalization of Cu together with the root activities demonstrate the different tolerance mechanism towards Cu toxicity of the two oat cultivars. In particular, cv. Fronteira seems to adopt an exclusion strategy based on accumulating Cu in the apoplasm and on the exudation of phenolic compounds. Thus, this cultivar could reduce the mobility and the consequent soil bioavailability of Cu playing an important role as phytostabilizer plant in intercropping systems in Cu-contaminated vineyards or orchards.

Evaluation of Copper-Contaminated Marginal Land for the Cultivation of Vetiver Grass (Chrysopogon zizanioides) as a Lignocellulosic Feedstock and its Impact on Downstream Bioethanol Production

Metal-contaminated soil could be sustainably used for biofuel feedstock production if the harvested biomass is amenable to bioethanol production. A 60-day greenhouse experiment was performed to evaluate (1) the potential of vetiver grass to phytostabilize soil contaminated with copper (Cu), and (2) the impact of Cu exposure on its lignocellulosic composition and downstream bioethanol production. Dilute acid pretreatment, enzymatic hydrolysis, and fermentation parameters were optimized sequentially for vetiver grass using response surface methodology (RSM). Results indicate that the lignocellulosic composition of vetiver grown on Cu-rich soil was favorably altered with a significant decrease in lignin and increase in hemicellulose and cellulose content. Hydrolysates produced from Cu exposed biomass achieved a significantly greater ethanol yield and volumetric productivity compared to those of the control biomass. Upon pretreatment, the hemicellulosic hydrolysate showed an increase in total sugars per liter by 204.7% of the predicted yield. After fermentation, 110% of the predicted ethanol yield was obtained for the vetiver grown on Cu-contaminated soil. By contrast, for vetiver grown on uncontaminated soil a 62.3% of theoretical ethanol yield was achieved, indicating that vetiver has the potential to serve the dual purpose of phytoremediation and biofuel feedstock generation on contaminated sites.

Assessing the effect of pyrolysis temperature on the molecular properties and copper sorption capacity of a halophyte biochar

The capacity of biochar to take up heavy metals from contaminated soil and water is influenced by the pyrolysis temperature. We have prepared three biochar samples from Jerusalem artichoke stalks (JAS) by pyrolysis at 300, 500 and 700 °C, denoted as JAS300, JAS500, and JAS700, respectively. A variety of synchrotron-based techniques were used to assess the effect of pyrolysis temperature on the molecular properties and copper (Cu) sorption capacity of the samples. The content of oxygen-containing functional groups in the biochar samples decreased, while that of aromatic structures and alkaline mineral components increased, with a rise in pyrolysis temperature. Scanning transmission X-ray microscopy indicated that sorbed Cu(II) was partially reduced to Cu(I), but this process was more evident with JAS300 and JAS700 than with JAS500. Carbon K-edge X-ray absorption near edge structure spectroscopy indicated that Cu(II) cations were sorbed to biochar via complexation and Cu-π bonding. With rising pyrolysis temperature, Cu(II)-complexation weakened while Cu-π bonding was enhanced. In addition, the relatively high ash content and pH of JAS500 and JAS700 facilitated Cu precipitation and the formation of langite on the surface of biochar. The results of this investigation will aid the conversion of halophyte waste to useable biochar for the effective remediation of Cu-contaminated soil and water.

Potential of vermicompost and limestone in reducing copper toxicity in young grapevines grown in Cu-contaminated vineyard soil

Foliar fungicide application in grapevines increases the content of heavy metals such as copper (Cu) in vineyard soils, which may reach phytotoxic levels. The application of soil amendments such as limestone and vermicompost may reduce Cu availability and phytotoxicity. The study aimed to assess growth, physiological status and changes in root morphology in young grapevines grown for 12 months in Cu-contaminated soil with and without the application of soil amendments. Samples of a Typic Hapludalf soil were collected in a vineyard with more than 30 years of cultivation. The following treatments were used: 1) control (without amendment), 2) application of organic vermicompost (86.7 g kg−1) and 3) application of limestone (3 Mg ha−1). Grapevines (Paulsen 1103 rootstock) were transplanted and grown for 12 months in PVC soil columns. We assessed parameters of growth, photosynthesis and root morphology. Grapevines grown in soil treated with limestone showed increased growth, dry matter yield and photosynthetic efficiency. The highest Cu concentrations in root tissue were found in grapevines grown in control soil. The application of vermicompost in this study did not alleviate Cu toxicity. Grapevines grown in soil treated with vermicompost showed high manganese (Mn) concentration in shoots before the winter pruning, reflecting the high Mn concentrations in soil solution that caused Mn phytotoxicity, resulting in plant death after the winter pruning. The vermicompost used in this study is not suitable for agronomic use. The use of limestone was an effective strategy to reduce Cu availability and phytotoxicity.

Influence of rice straw biochar on growth, antioxidant capacity and copper uptake in ramie (Boehmeria nivea L.) grown as forage in aged copper-contaminated soil

Copper (Cu) contamination in agricultural soil poses severe threats to living organisms, and possible ecofriendly solutions need to be considered for Cu immobilization, such as using biochar. A pot study was conducted to examine the effectiveness of biochar derived from rice straw (RSB) at various application rates (0, 2.5, 5 and 10% w/w) to mitigate possible risks of Cu solubility and its uptake by ramie (Boehmeria nivea L.) as forage. The plant growth parameters as well as soil chemical properties (pH, electrical conductivity and cation exchange capacity) notably improved with the increasing RSB application. Moreover, prominent reduction was observed in soil bioavailable Cu concentration by 96% with RSB application of 10% relative to control. In addition, Cu content in B. nivea roots, leaves and stems decreased by 60, 28 and 22%, respectively, for 10% RSB application. It was noted that chlorophyll content and gas exchange parameters in leaves were significantly higher at 10% RSB application than in control. Furthermore, 10% RSB resulted in a greater reduction in oxidative stress from the Cu in soil. Thus, soil amendment with RSB demonstrated positive results for Cu stabilization in aged Cu-contaminated soil, thereby reducing its accumulation and translocation in B. nivea and mitigating livestock feed security risks.

Assessment of bacterial communities in Cu-contaminated soil immobilized by a one-time application of micro-/nano-hydroxyapatite and phytoremediation for 3 years

Heavy metals contamination of soil has been considered as a global environmental problem, and consequently various soil amendments have been widely used in immobilization. Previous studies have reported that micro-/nano-hydroxyapatite (MHA/NHA) as a novel chemical material could alleviate soil acidity and reduce the bioavailability of heavy metals. However, the mechanism of soil microorganism responding to the application of MHA/NHA is little studied. Presently, an in-situ field experiment was conducted to determine the effects of MHA/NHA and the other three traditional amendments including alkali slag (AS), lime (L) and apatite (AP) on soil copper (Cu) bioavailability and dominate bacterial population. The results showed that the application of MHA/NHA effectively increased soil pH and decreased soil available Cu content, and showed the highest increasing effects on the activities of urease, catalase and acid phosphatase. Compared with the control, MHA/NHA significantly changed the soil bacterial community structure and increased the bacterial abundance and diversity. Besides, analysis of the dominate population showed that the application of MHA/NHA decreased the relative abundance of acidophiles and the indicator of soil degradation. Additionally, the relative abundance of potential plant growth promoting bacteria increased with the addition of MHA/NHA, which was confirmed by the characteristics (the ability of producing indole acetic acid and siderophore) of bacterial strains. These results suggested that these dominate bacterial populations with significant changes may be regarded as the biomarkers for the recovery of soil ecological environment, which provides a theoretical basis for the ecological evaluation of MHA/NHA.

Mycorrhizal Inoculation Differentially Affects Grapevine's Performance in Copper Contaminated and Non-contaminated Soils.

Plant inoculation with arbuscular mycorrhizal fungi (AMF) is increasingly employed to enhance productivity and sustainability in agricultural ecosystems. In the present study, the potential benefits of AMF inoculation on young grapevines replanted in pots containing vineyard soil with high Cu concentration were evaluated. For this purpose, one-year-old cv. Touriga Nacional grapevines grafted onto 1103P rootstocks were further inoculated with Rhizoglomus irregulare or Funneliformis mosseae, or left non-inoculated, and maintained in a sterilized substrate under greenhouse conditions for three months. After this time, half of the plants were transplanted to containers filled with an Arenosol from a vineyard which had been artificially contaminated or not with 300 mg kg−1 of Cu. At the end of the growing season, soil nutrient concentration, soil dehydrogenase activity and mycorrhizal colonization rate were analyzed. Grapevine performance was assessed by measuring several vegetative growth and physiological parameters as well as nutrient concentrations in leaves and roots. In the non-contaminated soil, R. irregulare- and F. mosseae-inoculated plants had significantly greater root biomass than the non-inoculated ones. However, the opposite effect was observed in the Cu-contaminated soil, where non-inoculated plants performed better regarding shoot and root development. Concerning nutrient levels, an increase in Cu, Mg and Mn concentrations was observed in the roots of plants growing in the contaminated soil, although only Mn was translocated to leaves. This led to a large increase in leaf Mn concentrations, which was significantly higher in non-inoculated and F. mosseae- inoculated plants than in the R. irregulare- inoculated ones. Copper contamination induced a general decrease in leaf N, P and Fe concentrations as well as chlorosis symptoms. The largest decrease in N and P was observed in F. mosseae- inoculated plants, with 73 and 31.2%, respectively. However, these plants were the ones with the least decrease in Fe concentration (10% vs. almost 30% in the other two inoculation treatments). In conclusion, this study indicates that soil Cu levels can modify the outcome of AMF inoculations in young grapevines, disclosing new AMF-plant associations potentially relevant in vineyards with a tradition of Cu-based fungicide application.

Effects of copper on the growth, antioxidant enzymes and photosynthesis of spinach seedlings

Examination of plants with strong Cu tolerance and an understanding of their Cu-tolerance mechanisms are of considerable significance for the remediation of Cu-contaminated soil. Although spinach may be a plant with strong Cu tolerance, the threshold of Cu tolerance in this plant and its physiological response mechanisms to Cu are still unclear. In this study, we examined that the effects of different Cu concentrations on the growth parameters, antioxidant enzyme activities, and photosynthesis of spinach seedlings. The results showed that when treated with a low Cu concentration (100 mg L−1 CuSO4), the biomass of spinach seedlings increased, whereas the MDA content, the activities of antioxidant enzymes, Pn, gs and Tr were not significantly different from those in the control (P > 0.05), and Y(II), qP reached their maximum values, indicating that a low Cu concentration (100 mg L−1 CuSO4) had minimal negative effects on the life activities of spinach seedlings. In contrast, when treated with high Cu concentrations (800–1000 mg L−1 CuSO4), the total biomass of spinach seedlings was markedly decreased, the MDA contents increased, antioxidant enzyme activities initially increased and then decreased to varying degrees, the contents of chlorophyll, Pn, Tr, Fv/Fm, qP, NPQ, and Y(II) were all decreased. However the growth of spinach did not terminate, implying that the lethal threshold concentration of Cu for spinach is greater than 1000 mg L−1 CuSO4 used in this study. In summary, spinach exhibits a high tolerance to Cu and can be considered as an alternative plant for the remediation of Cu-contaminated soils.

Co-Amendment of S and Si Alleviates Cu Toxicity in Rice (Oryza Sativa L.) Grown on Cu-Contaminated Paddy Soil.

With irrigation using waste water, application of sewage sludge, and development of mine exploration, copper (Cu) contamination in some paddy fields has become increasingly serious. A greenhouse pot experiment was conducted using a factorial design with three sulfur (S) application rates (i.e., 0, 0.013, and 0.026 g S kg−1 soil) and three silicon (Si) application rates (i.e., 0, 0.05, and 0.1 g Si kg−1 soil) to test the effect of co-amendment of S and Si on alleviating Cu contamination in paddy soil. There were significant interaction effects between S and Si on soil Cu speciation and Cu uptake by rice plants (except brown rice). Sulfur addition decreased the content of soil-exchangeable Cu, whereas Si addition decreased the content of soil-reducible Cu, suggesting that co-amendment of S and Si generally reduced Cu availability. Copper was biominimized in the soil-rice plant system and rice root had the greatest Cu concentration (163–285 mg kg−1). Co-amendment of S and Si decreased the translocation of Cu from soil to rice root, possibly due to decreased soil Cu mobility and enhancement of the formation of iron plaque on rice root. Co-amendment of S-Si at a rate of 0.013 (S)–0.1 (Si) g kg−1 soil, respectively, was the optimal among all treatments.

Growth, biochemical response and nutritional status of Angico-Vermelho (Parapiptadenia rigida (Bentham) Brenan) under the application of soil amendment in Cu-contaminated soil

Forest species Angico-Vermelho (Parapiptadenia rigida (Bentham) Brenan) is an alternative for the revegetation of areas contaminated with high levels of heavy metals such as copper (Cu). However, excess Cu may cause toxicity to plants, which is why the use of soil amendments can facilitate cultivation by reducing the availability of Cu in the soil. The aim of this study was to assess how the use of amendment can contribute to growth and nutritional status as well as reduce oxidative stress in Angico-Vermelho grown in Cu-contaminated soil. Samples of a Typic Hapludalf soil containing high Cu content were used for the application of four amendments (limestone, organic compost, Ca silicate and zeolite), in addition to a control treatment. The treatments were arranged in a completely randomized design, with four replicates. The use of amendments decreased Cu content available in soil and contributed to improve both plant nutritional status and its antioxidant response expressed by enzymatic activity. The application of the amendments, especially zeolite and Ca silicate, increased dry matter yield of Angico-Vermelho. Thus, the results presented here suggest that the use of amendments contributes to improving Cu-contaminated soils and favors revegetation with Angico-Vermelho.

Copper phytoextraction and phytostabilization potential of wild plant species growing in the mine polluted areas of Armenia

The assessment of phytoremediation potential of wild plant species in areas polluted by metals is very important, as it will enable the use of eco-friendly remediation methods in these territories. Sixteen native wild plant species growing in Cu-contaminated soils of a mining region in Armenia were investigated to reveal their phytoremediation potential. In roots (dry weight) of dominant plant species in this area the content of copper varied between 55 mg/kg ( Hypericum perforatum ) and 775 mg/kg ( Thymus kotschyanus ), and in shoots of plants in the range from 33 mg/kg ( Teucrium orientale ) to 243 mg/kg ( Phleum pratense ). Since the Cu accumulation capability of plants depends both on physiological peculiarities of the plants and on the content of Cu bioavailable in the soil, the studies were carried out in this direction and it was found out that the high contents of soil organic matter (SOM) and clay in the soil facilitated the decrease of the ratio Cu bioavailable /Cu total and as a result, the decrease of Cu accumulation capability of plants. Thymus kotschyanus , Phleum pratense and Achillea millefolium had the highest phytostabilization potential from all studied species due to high bioconcentration factors of their roots (BCF root ) and low translocation factors (TF) registered in these plants.

Intercropping of young grapevines with native grasses for phytoremediation of Cu-contaminated soils

Intercropping may be a strategy for phytoremediation of vineyard soils with high copper (Cu) content. The study aimed to evaluate the contribution of South American native grasses in limiting Cu availability and toxicity in soils grown with grapevines. The soil used in the experiment was collected in natural grassland with no history of cultivation. The samples were air-dried; acidity, P and K levels were corrected and samples were then incubated. We used three Cu levels - natural content (Dose 0) and the addition of 40 and 80 mg Cu kg−1 of soil (Dose 40 and 80). At each Cu dose, grapevine was grown in three cropping treatments: monocropping, intercropping with Paspalum plicatulum and intercropping with Axonopus affinis. In intercropping, two grass seedlings were transplanted into each experimental unit 35 days prior to the transplanting of the grapevines. The soil solution was sampled and ionic speciation was carried out. At 70 days after planting, we sampled the grapevines to determine dry matter, morphological parameters and nutrient concentration in the roots and shoots. Intercropping young grapevines with Paspalum plicatulum and Axonopus affinis was efficient in promoting the growth of young grapevines at moderate and low levels of Cu contamination by reducing Cu bioavailability. This indicates that maintaining native grasses in young vineyards is an effective strategy for phytoremediating Cu-contaminated soils and obtaining a grape production system with reduced interventions in the native environment, in addition to contributing to soil protection and nutrient cycling.

Phytomanagement and Remediation of Cu-Contaminated Soils by High Yielding Crops at a Former Wood Preservation Site: Sunflower Biomass and Ionome

This long-term field trial aimed at remediating a Cu-contaminated soil to promote crop production and soil functions at a former wood preservation site. Twenty-eight field plots with total topsoil Cu in the 198-1,169 mg kg-1 range were assessed. Twenty-four plots (OMDL) were amended in 2008 with a compost (made of pine bark chips and poultry manure, OM, 5% w/w) and dolomitic limestone (DL, 0.2%), and thereafter annually phytomanaged with a sunflower-tobacco crop rotation. In 2013, one untreated plot (UNT) was amended with a green waste compost (GW, 5%) whereas 12 former OMDL plots received a second compost dressing using this green waste compost (OM2DL, 5%). In 2011, one plot was amended with the Carmeuse basic slag (CAR, 1%) and another plot with a P-spiked Linz-Donawitz basic slag (PLD, 1%). Thus six soil treatments, i.e., UNT, OMDL, OM2DL, GW, CAR, and PLD, were cultivated in 2016 with sunflower (Helianthus annuus L. cv Ethic). Shoots were harvested and their ionome analyzed. At high soil Cu contamination, the 1M NH4NO3-extractable vs. total soil Cu ratio ranked in decreasing order: Unt (2.35) > CAR (1.02), PLD (0.83) > GW (0.58), OMDL (0.44), OM2DL (0.37), indicating a lower Cu extractability in the compost-amended plots. All amendments improved the soil nutrient status and the soil pH, which was slightly acidic in the UNT soil. Total organic C and N and extractable P contents peaked in the OM2DL soils. Both OMDL and OM2DL treatments led to higher shoot DW yields and Cu removals than the GW, CAR, and PLD treatments. Shoot DW yields decreased as total topsoil Cu rose in the OMDL plots, on the contrary to the OM2DL plots, demonstrating the benefits to repeat compost application after 5 years. Shoot Cu concentrations notably of OMDL and OM2DL plants fitted into their common range and can be used by biomass processing technologies and oilseeds as well. In overall, there is a net gain in soil physico-chemical properties and underlying soil functions

Rhizophagus Clarus and Phosphorus in Crotalaria juncea: Growth, Glomalin Content and Acid Phosphatase Activity in a Copper-Contaminated Soil

ABSTRACT Crotalaria juncea is used as plant cover in grape vineyards in Brazil, which usually present soils with high copper (Cu) levels due to the application of Cu-based phyto-sanitary products. Under this condition an increase growth and cover of C. juncea is needed to improve the phytoremediation processes in those soils. Some alternatives to achieve this condition is the inoculation with arbuscular mycorrhizal fungi (AMF), which has demonstrated an important increase of plant growth in Cu-contaminated soils at different soil [...]

Glutathione S-transferases modulate Cu tolerance in Oryza sativa

Along with the widespread use of copper (Cu)-based fungicides and fertilizers, Cu-contaminated soil has been a major issue resulting in the loss of crop yield. To improve Cu tolerance in Oryza sativa, we previously carried out a screen of Cu sensitivity and obtained the Cu tolerant (B1139) and the Cu sensitive (B1195) rice varieties. Here, by analyzing an RNA-Seq transcriptome profile of root tips, 20 glutathione S-transferase (GST) genes were differently expressed between the Cu tolerant B1139 and the Cu sensitive B1195 when exposed to excess Cu2+. A more rapid decrease of root GST activity occurred in Cu-treated B1195 than in Cu-treated B1139, suggesting that different GST activity might affect Cu tolerance in rice. This is further supported by the observation of Cu hypersensitivity in rice seedlings deficient of the most abundant GST protein, OsGSTF2, demonstrating that GST proteins modulate Cu tolerance in rice. This, at least partially, leads to different Cu tolerance between B1139 and B1195. Furthermore, homologous modeling showed that a putative Cu2+ binding site, OsGSTF2H41 locates at the active center of GST enzyme. In vitro analysis demonstrated that Cu2+ interferes with the GST activity of OsGSTF2 protein, indicating that GST activity reduction by Cu2+ might be the binding of Cu2+ to the OsGSTF2H41 site. Consistently, amino acid replacement of histidine 41 with glycine (OsGSTF2H41G) resulted in decreased GST activity of OsGSTF2. Additionally, ectopic expression of OsGSTF2 enhances Cu tolerance in Escherichia coli, suggesting that the GST proteins might alleviate Cu toxicity by the binding of free Cu2+ in cells.

Rhizobium inoculation enhances copper tolerance by affecting copper uptake and regulating the ascorbate-glutathione cycle and phytochelatin biosynthesis-related gene expression in Medicago sativa seedlings

Despite numerous reports that legume-rhizobium symbiosis alleviates Cu stress in plants, the possible roles of legume-rhizobium symbiosis and the regulatory mechanisms in counteracting Cu toxicity remain unclear. Here, Sinorhizobium meliloti CCNWSX0020 was used for analyzing the effects of rhizobium inoculation on plant growth in Medicago sativa seedlings under Cu stress. Our results showed that rhizobium inoculation alleviated Cu-induced growth inhibition, and increased nitrogen concentration in M. sativa seedlings. Moreover, the total amount of Cu uptake in inoculated plants was significantly increased compared with non-inoculated plants, and the increase in the roots was much higher than that in the shoots, thus decreasing the transfer coefficient and promoting Cu phytostabilization. Cu stress induced lipid peroxidation and reactive oxygen species production, but rhizobium inoculation reduced these components’ accumulation through altering antioxidant enzyme activities and regulating ascorbate-glutathione cycles. Furthermore, legume-rhizobium symbiosis regulated the gene expression involved in antioxidant responses, phytochelatin (PC) biosynthesis, and metallothionein biosynthesis in M. sativa seedlings under Cu stress. Our results demonstrate that rhizobium inoculation enhanced Cu tolerance by affecting Cu uptake, regulating antioxidant enzyme activities and the ascorbate-glutathione cycle, and influencing PC biosynthesis-related gene expression in M. sativa. The results provide an efficient strategy for phytoremediation of Cu-contaminated soils.

An assessment of copper-contaminated soil using Kenaf (Hibiscus cannabinus l.) as a phytoaccumulator

A pot experiment was conducted to study the potentials of Kenaf (Hibiscus cannabinus L.) as a Copper (Cu) phytoaccumulator at different levels of artificial contamination. Copper was applied as cupric sulphate (CuSO4.5H2O) and the treatments were 0, 50, 100 and 150 mg kg-1 of Cu. Kenaf was grown in each of the treated pots for 10 weeks, following which, leaf, stem and root samples were collected and analysed for Cu contents. The soil was analysed for physicochemical properties [viz. pH, electrical conductivity (EC), organic carbon (OC), exchangeable acidity (EA), exchangeable cations (Ca2+, Mg2+, Na+, K+) and extractable and total Cu contents) before treatments/contamination and after harvesting. Kenaf showed symptoms of toxicity at 100 and 150 mg kg-1 treatments of Cu. It was observed that, compared with the levels of contamination of Cu, the concentration in kenaf was generally insignificant; thus, the concentration reduction in the soil at one cycle of cropping may not berealistic.  Copper treatments at different levels significantly changed soil pH and EC. Therefore, more cycles of growth are needed to effectively remediate Cu-contaminated soils using Kenaf as Cu phytoaccumulator.