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Use of sunflower (Helianthus annuus L.) for Cu phytoextraction and oilseed production on Cu-contaminated topsoils was investigated in afield trial at a former wood preservation site. Six commercial cultivars and two mutant lines were cultivated in plots with and without the addition of compost (5% w/w) and dolomitic limestone (0.2% w/w). Total soil Cu ranged from 163 to 1170 mg kg(-1). In soil solutions, Cu concentration varied between 0.16-0.93 mg L(-1). The amendment increased soil pH, reduced Cu exposure and promoted sunflower growth. Stem length, shoot and capitulum biomasses, seed yield, and shoot and leaf Cu concentrations were measured. At low total soil Cu, shoot Cu mineralomass was higher in commercial cultivars, Le., Salut, Energic, and Countri, whereas competition and shading affected morphological traits of mutants. Based on shoot yield (7 Mg DW ha(-1)) and Cu concentration, the highest removal was 59 g Cu ha(-1). At high total soil Cu, shoot Cu mineralomass peaked for mutants (e.g., 52 g Cu ha(-1) for Mutant 1 line) and cultivars Energic and Countri. Energic seed yield (3.9 Mg air-DW ha(-1)) would be sufficient to produce oil Phenotype traits and shoot Cu removal depended on sunflower types and Cu exposure.

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 NH 4 NO 3-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 Mench et al. Phytomanagement of Cu-Contaminated Soils processing technologies and oilseeds as well. In overall, there is a net gain in soil physico-chemical properties and underlying soil functions. HIGHLIGHTS-Compost incorporated into Cu-contaminated soils improves the sunflower growth.-Soil organic matter increases in compost-amended soils.-Extractable soil Cu decreases in compost-amended soils.-Shoot Cu removal by sunflower reaches 26-88 g Cu ha −1 year −1 .

Plant species, spatial variability in plant diversity and vegetation cover were recorded at a French timber treatment site with Cu-contaminated soils (65-2600 mg/kg). Shoot biomass, shoot Cu concentration and accumulation were determined for each plant species found on 168 quadrats with increasing total Cu in soil and soil solution. A total of 91 species occurred on the site including four considered as invasive (Cyperus eragrostis, Phytolacca americana, Senecio inaequidens, and Sporobolus indicus). Species richness, Shannon index, vegetation cover and plant biomass decreased as soil Cu increased. At low soil Cu, members of the Poaceae were most frequent followed by Fabaceae, Rosaceae, and Asteraceae. At high soil Cu, Poaceae were again most frequent. Species known to form Cu-tolerant populations, i.e. Agrostis capillaris, A. stolonifera and Rumex acetosella were present. Shoot Cu concentration and accumulation were higher in plants growing in the most contaminated soils. At 2142 mg Cu/kg soil, shoot Cu accumulation peaked at 6 mg Cu/m² in A. capillaris, and its shoot Cu concentration (364 mg Cu/kg dry weight) exceeded the fodder Cu threshold for domestic livestock. In less Cu-contaminated soils some candidates were identified for sustainable phytoremediation with a potential financial return.

Trace elements (TEs) availability, biochemical activity and functional gene diversity was studied in a Cu-contaminated soil, revegetated for six years with a mixed stand of willow, black poplar, and false indigo-bush, and amended or not with compost plus dolomitic limestone (OMDL). The OMDL amendment significantly reduced Cu and As availability and soil toxicity, and increased the biochemical activity and microbial functional diversity assessed with the GEOCHIP technique, as compared to the unamended soil (Unt). The OMDL soil showed significantly higher abundance of 25 functional genes involved in decomposition organic compounds, and 11, 3 and 11 functional genes involved in the N, P and S biogeochemical cycles. Functional gene abundance was positively correlated with nutrient contents but negatively correlated with Cu availability and soil toxicity. The abundance of microbial functional genes encoding for resistance to various TEs also increased, possibly due to the microbial proliferation and lower Cu exposure in the presence of high total soil Cu concentration. Genes encoding for antibiotic resistance due to the co-occurrence of TEs and antibiotic resistant genes on genetic mobile elements. Overall, phytomanagement confirmed its potential to restore the biological fertility and diversity of a severely Cu-contaminated soil, but the increase of TEs and antibiotic resistant gene abundances deserve attention in future studies.

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Differential expression of soluble proteins was explored in roots of metallicolous (M) and non-M (NM) plants of Agrostis capillaris L. exposed to increasing Cu to partially identify molecular mechanisms underlying higher Cu tolerance in M plants. Plants were cultivated for 2 months on perlite with a CuSO4 (1-30 μM) spiked-nutrient solution. Soluble proteins extracted by the trichloroacetic acid/acetone procedure were separated with 2DE (linear 4-7 pH gradient). After Coomassie Blue staining and image analysis, 19 proteins differentially expressed were identified using LC-MS/MS and Expressed Sequence Tag (ESTs) databases. At supra-optimal Cu exposure (15-30 μM), glycolysis was likely altered in NM roots with increased production of glycerone-P and methylglyoxal based on overexpression of triosephosphate isomerase and fructose bisphosphate aldolase. Changes in tubulins and higher expressions of 5-methyltetrahydropteroyltriglutamatehomocysteine methyltransferase and S-adenosylmethionine synthase underpinned impacts on the cytoskeleton and stimulation of ethylene metabolism. Increased l-methionine and S-adenosylmethionine amounts may also facilitate production of nicotianamine, which complexes Cu, and of l-cysteine, needed for metallothioneins and GSH. In M roots, the increase of [Cu/Zn] superoxide dismutase suggested a better detoxification of superoxide, when Cu exposure rose. Higher Cu-tolerance of M plants would rather result from simultaneous cooperation of various processes than from a specific mechanism.