A multidisciplinary and structured investigation of three suspected clusters of transverse upper limb reduction defects in France.

INTRODUCTION: Between 2019-2021, facing public concern, a scientific expert committee (SEC) reanalysed suspected clusters of transverse upper limb reduction defects (TULRD) in three administrative areas in France, where initial investigations had not identified any risk exposure. We share here the national approach we developed for managing suspicious clusters of the same group of congenital anomalies occurring in several areas. METHODS: The SEC analysed the medical records of TURLD suspected cases and performed spatiotemporal analyses on confirmed cases. If the cluster was statistically significant and included at least three cases, the SEC reviewed exposures obtained from questionnaires, environmental databases, and a survey among farmers living near to cases' homes concerning their plant product use. RESULTS: After case re-ascertainment, no statistically significant cluster was observed in the first administrative areas. In the second area, a cluster of four children born in two nearby towns over two years was confirmed, but as with the initial investigations, no exposure to a known risk factor explaining the number of cases in excess was identified. In the third area, a cluster including just two cases born the same year in the same town was confirmed. DISCUSSION: Our experience highlights that in the event of suspicious clusters occurring in different areas of a country, a coordinated and standardised approach should be preferred.

Limited effects of crop foliar Si fertilization on a marginal soil under a future climate scenario.

Growing crops on marginal lands is a promising solution to alleviate the increasing pressure on agricultural land in Europe. Such crops will however be at the same time exposed to increased drought and pathogen prevalence, on already challenging soil conditions. Some sustainable practices, such as Silicon (Si) foliar fertilization, have been proposed to alleviate these two stress factors, but have not been tested under controlled, future climate conditions. We hypothesized that Si foliar fertilization would be beneficial for crops under future climate, and would have cascading beneficial effects on ecosystem processes, as many of them are directly dependent on plant health. We tested this hypothesis by exposing spring barley growing on marginal soil macrocosms (three with, three without Si treatment) to 2070 climate projections in an ecotron facility. Using the high-capacity monitoring of the ecotron, we estimated C, water, and N budgets of every macrocosm. Additionally, we measured crop yield, the biomass of each plant organ, and characterized bacterial communities using metabarcoding. Despite being exposed to water stress conditions, plants did not produce more biomass with the foliar Si fertilization, whatever the organ considered. Evapotranspiration (ET) was unaffected, as well as water quality and bacterial communities. However, in the 10-day period following two of the three Si applications, we measured a significant increase in C sequestration, when climate conditions where significantly drier, while ET remained the same. We interpreted these results as a less significant effect of Si treatment than expected as compared with literature, which could be explained by the high CO2 levels under future climate, that reduces need for stomata opening, and therefore sensitivity to drought. We conclude that making marginal soils climate proof using foliar Si treatments may not be a sufficient strategy, at least in this type of nutrient-poor, dry, sandy soil.

Plant testing with hemp and miscanthus to assess phytomanagement options including biostimulants and mycorrhizae on a metal-contaminated soil to provide biomass for sustainable biofuel production.

The need of biofuels from biomass, including sustainable aviation fuel, without using agricultural land dedicated to food crops, is in constant demand. Strategies to intensify biomass production using mycorrhizal fungi, biostimulants and their combinations could be solutions for improving the cultivation of lignocellulosic plants but still lack well-established validation on metal-contaminated soils. This study aimed to assess the yield of Miscanthus x giganteus J.M. Greef & Deuter and Cannabis sativa L. grown on a metal-contaminated agricultural soil (11 mg Cd, 536 mg Pb and 955 mg Zn kg-1) amended with biostimulants and/or arbuscular mycorrhizal fungi, and the shoot Cd, Pb and Zn uptake. A pot trial was carried out with soil collected from a field near a former Pb/Zn smelter in France and six treatments: control (C), protein hydrolysate (a mixture of peptides and amino acids, PH), humic/fulvic acids (HFA), arbuscular mycorrhizae fungi (AMF), PH combined with AMF (PHxAMF), and HFA combined with AMF (HFAxAMF). Metal concentrations in the soil pore water (SPW), pH and electrical conductivity were measured over time. Miscanthus and hemp shoots were harvested on day 90. Both PH and PHxAMF treatments increased SPW Cd, Pb, and Zn concentrations (e.g. by 26, 1.9, and 22.9 times for miscanthus and 9.7, 4.7, and 19.3 times for hemp in the PH and PHxAMF treatments as compared to the control one, respectively). This led to phytotoxicity and reduced shoot yield for miscanthus. Conversely, HFA and HFAxAMF treatments decreased SPW Cd and Zn concentrations, increasing shoot yields for hemp and miscanthus. Shoot Cd, Pb, and Zn uptakes peaked for PH and PHxAMF hemp plants (in µg plant-1, Cd: 310-334, Pb: 34-38, and Zn: 232-309 for PHxAMF and PH, respectively), while lowest values occurred for PH miscanthus plants mainly due to low shoot yield. Overall, this study suggested that humic/fulvic acids can be an effective biostimulant for increasing shoot biomass production in a metal-contaminated soil. These results warrant further investigations of the HFAxAMF in field trials.

Drivers to improve metal(loid) phytoextraction with a focus on microbial degradation of dissolved organic matter in soils.

Bioaugmentation of soils can increase the mobilization of metal(loid)s from the soil-bearing phases. However, once desorbed, these metal(loid)s are mostly complexed to the dissolved organic matter (DOM) in the soil solution, which can restrict their availability to plants (roots mainly taking up the free forms) and then the phytoextraction performances. Firstly the main drivers influencing phytoextraction are reminded, then the review focuses on the DOM role. After having reminding the origin, the chemical structure and the lability of DOM, the pool of stable DOM (the most abundant in the soil) most involved in the complexation of metal(loid)s is addressed in particular by focusing on carboxylic and/or phenolic groups and factors controlling metal(loid) complexation with DOM. Finally, this review addresses the ability of microorganisms to degrade metal(loid)-DOM complexes as an additional lever for increasing the pool of free metal(loid) ions, and then phytoextraction performances, and details the origin of microorganisms and how they are selected. The development of innovative processes including the use of these DOM-degrading microorganisms is proposed in perspectives.

This review focuses on the available drivers to increase the pool of free (i.e. phytoavailable) metal(loid)s in the soil solution, with a specific focus on the ability of microorganisms to degrade dissolved organic matter for enriching this pool, and then to substantially improve phytoextraction performance.

Field assessment of organic amendments and spring barley to phytomanage a Cu/PAH-contaminated soil.

The INTENSE project, supported by the EU Era-Net Facce Surplus, aimed at increasing crop production on marginal land, including those with contaminated soils. A field trial was set up at a former wood preservation site to phytomanage a Cu/PAH-contaminated sandy soil. The novelty was to assess the influence of five organic amendments differing in their composition and production process, i.e. solid fractions before and after biodigestion of pig manure, compost and compost pellets (produced from spent mushroom substrate, biogas digestate and straw), and greenwaste compost, on Cu availability, soil properties, nutrient supply, and plant growth. Organic amendments were incorporated into the soil at 2.3% and 5% soil w/w. Total soil Cu varied from 179 to 1520 mg kg-1, and 1 M NH4NO3-extractable soil Cu ranged from 4.7 to 104 mg kg-1 across the 25 plots. Spring barley (Hordeum vulgare cv. Ella) was cultivated in plots. Changes in physico-chemical soil properties, shoot DW yield, shoot ionome, and shoot Cu uptake depending on extractable soil Cu and the soil treatments are reported. Shoot Cu concentration varied from 45 ± 24 to 140 ± 193 mg kg DW-1 and generally increased with extractable soil Cu. Shoot DW yield, shoot Cu concentration, and shoot Cu uptake of barley plants did not significantly differ across the soil treatments in year 1. Based on soil and plant parameters, the effects of the compost and pig manure treatments were globally discriminated from those of the untreated, greenwaste compost and digested pig manure treatments. Compost and its pellets at the 5% addition rate promoted soil functions related to primary production, water purification, and soil fertility, and the soil quality index.

Biomass partitioning of plants under soil pollution stress.

Polluted sites are ubiquitous worldwide but how plant partition their biomass between different organs in this context is unclear. Here, we identified three possible drivers of biomass partitioning in our controlled study along pollution gradients: plant size reduction (pollution effect) combined with allometric scaling between organs; early deficit in root surfaces (pollution effect) inducing a decreased water uptake; increased biomass allocation to roots to compensate for lower soil resource acquisition consistent with the optimal partitioning theory (plant response). A complementary meta-analysis showed variation in biomass partitioning across published studies, with grass and woody species having distinct modifications of their root: shoot ratio. However, the modelling of biomass partitioning drivers showed that single harvest experiments performed in previous studies prevent identifying the main drivers at stake. The proposed distinction between pollution effects and plant response will help to improve our knowledge of plant allocation strategies in the context of pollution.

Relaunch cropping on marginal soils by incorporating amendments and beneficial trace elements in an interdisciplinary approach.

In the EU and world-wide, agriculture is in transition. Whilst we just converted conventional farming imprinted by the post-war food demand and heavy agrochemical usage into integrated and sustainable farming with optimized production, we now have to focus on even smarter agricultural management. Enhanced nutrient efficiency and resistance to pests/pathogens combined with a greener footprint will be crucial for future sustainable farming and its wider environment. Future land use must embrace efficient production and utilization of biomass for improved economic, environmental, and social outcomes, as subsumed under the EU Green Deal, including also sites that have so far been considered as marginal and excluded from production. Another frontier is to supply high-quality food and feed to increase the nutrient density of staple crops. In diets of over two-thirds of the world's population, more than one micronutrient (Fe, Zn, I or Se) is lacking. To improve nutritious values of crops, it will be necessary to combine integrated, systems-based approaches of land management with sustainable redevelopment of agriculture, including central ecosystem services, on so far neglected sites: neglected grassland, set aside land, and marginal lands, paying attention to their connectivity with natural areas. Here we need new integrative approaches which allow the application of different instruments to provide us not only with biomass of sufficient quality and quantity in a site specific manner, but also to improve soil ecological services, e.g. soil C sequestration, water quality, habitat and soil resistance to erosion, while keeping fertilization as low as possible. Such instruments may include the application of different forms of high carbon amendments, the application of macro- and microelements to improve crop performance and quality as well as a targeted manipulation of the soil microbiome. Under certain caveats, the potential of such sites can be unlocked by innovative production systems, ready for the sustainable production of crops enriched in micronutrients and providing services within a circular economy.

Potential ability of tobacco (Nicotiana tabacum L.) to phytomanage an urban brownfield soil.

The ability of tobacco (Nicotiana tabacum L. cv. Badischer Geudertheimer) for phytomanaging and remediating soil ecological functions at a contaminated site was assessed with a potted soil series made by fading an uncontaminated sandy soil with a contaminated sandy soil from the Borifer brownfield site, Bordeaux, SW France, at the 0%, 25%, 50%, 75%, and 100% addition rates. Activities of sandblasting and painting with metal-based paints occurred for decades at this urban brownfield, polluting the soil with metal(loid)s and organic contaminants, e.g., polycyclic aromatic hydrocarbons, in addition to past backfilling. Total topsoil metal(loid)s (e.g., 54,700 mg Zn and 5060 mg Cu kg-1) exceeded by seven- to tenfold the background values for French sandy soils, but the soil pH was 7.9, and overall, the 1M NH4NO3 extractable soil fractions of metals were relatively low. Leaf area, water content of shoots, and total chlorophyll (Chl) progressively decreased with the soil contamination, but the Chl fluorescence remained constant near its optimum value. Foliar Cu and Zn concentrations varied from 17.8 ± 4.2 (0%) to 27 ± 5 mg Cu kg-1 (100%) and from 60 ± 15 (0%) to 454 ± 53 mg Zn kg-1 (100%), respectively. Foliar Cd concentration peaked up to 1.74 ± 0.09 mg Cd kg-1, and its bioconcentration factor had the highest value (0.2) among those of the metal(loid)s. Few nutrient concentrations in the aboveground plant parts decreased with the soil contamination, e.g., foliar P concentration from 5972 ± 1026 (0%) to 2861 ± 334 mg kg-1 (100%). Vulnerability to drought-induced embolism (P50) did not differ for the tobacco stems across the soil series, whereas their hydraulic efficiency (Ks) declined significantly with increasing soil contamination. Overall, this tobacco cultivar grew relatively well even in the Borifer soil (100%), keeping its photosynthetic system healthy under stress, and contaminant exposure did not increase the vulnerability of the vascular system to drought. This tobacco had a relevant potential to annually phytoextract a part of the bioavailable soil Zn and Cd, i.e., shoot removals representing here 8.8% for Zn and 43.3% for Cd of their 1M NH4NO3 extractable amount in the potted Borifer soil.

Cu phytoextraction and biomass utilization as essential trace element feed supplements for livestock.

Copper (Cu), as an essential element, is added to animal feed to stimulate growth and prevent disease. The forage crop alfalfa (Medicago sativa L.) produced during Cu phytoextraction may be considered a biofortified crop to substitute the Cu feed additives for livestock production, beneficially alleviating Cu contamination in soils and reducing its input into agriculture systems. To assess this, alfalfa was grown in three similar soils with different Cu levels, i.e., 11, 439 and 779 mg kg-1 for uncontaminated soil (A), moderately Cu-contaminated soil (B) and highly Cu-contaminated soil (C), respectively. EDDS (Ethylenediamine-N,N'-disuccinic acid) was applied to the soils seven days before the first cutting at four rates (0, 0.5, 2 and 5 mmol kg-1) to enhance bioavailable Cu uptake. Alfalfa grew well in soils A and B but not in the highly Cu-contaminated soil. After applying EDDS, a significant biomass reduction of the first cutting shoot was only observed with 5 mmol kg-1 EDDS in the highly Cu-contaminated soil, with a 45% (P < 0.05) decrease when compared to the control. Alfalfa grown in the three soils gradually wilted after the first cutting with 5 mmol kg-1 EDDS, and Cu concentrations in the first cutting shoot were augmented strongly, by 250% (P < 0.05), 3500% (P < 0.05) and 6700% (P < 0.05) compared to the controls, respectively. Cu concentrations in alfalfa shoots were found to be higher in this study than in some fodder plants and further augmented in soils with higher Cu levels and with EDDS application. These findings suggest that alfalfa grown on clean soils or soils with up to 450 mg Cu kg-1 (with appropriate EDDS dosages) has the potential to be considered as a partial Cu supplementation for livestock. This research laid the foundation for the integration between Cu-phytoextraction and Cu-biofortification for livestock.

Water and soil contaminated by arsenic: the use of microorganisms and plants in bioremediation.

Owing to their roles in the arsenic (As) biogeochemical cycle, microorganisms and plants offer significant potential for developing innovative biotechnological applications able to remediate As pollutions. This possible use in bioremediation processes and phytomanagement is based on their ability to catalyse various biotransformation reactions leading to, e.g. the precipitation, dissolution, and sequestration of As, stabilisation in the root zone and shoot As removal. On the one hand, genomic studies of microorganisms and their communities are useful in understanding their metabolic activities and their interaction with As. On the other hand, our knowledge of molecular mechanisms and fate of As in plants has been improved by laboratory and field experiments. Such studies pave new avenues for developing environmentally friendly bioprocessing options targeting As, which worldwide represents a major risk to many ecosystems and human health.

LONG-TERM stability of arsenic in iron amended contaminated soil.

This study aimed at elucidating the long-term efficiency of soil remediation where chemical stabilization of arsenic (As) contaminated soil using zerovalent iron (Fe) amendments was applied. A combination of chemical extraction and extended X-Ray absorption fine structure (EXAFS) spectroscopy technique was applied on soils collected from five laboratory and field experiments in Sweden and France. All soils were treated with 1 wt% of zerovalent Fe grit 2-15 years prior to the sampling. The results indicate that all studied soils, despite the elapsed time since their amendment with Fe grit, had substantial amounts of ferrihydrite and/or lepidocrocite. These metastable and the most reactive Fe (oxyhydr)oxides (mainly ferrihydrite) were still present in substantial amounts even in the soil that was treated 15 years prior to the sampling and contributed most to the As immobilisation in the amended soils. This increases confidence in the long-term efficiency of As immobilisation using zerovalent Fe amendments. Both applied methods, sequential extraction and EXAFS, were in line for most of the samples in terms of their ability to highlight As immobilisation by poorly crystalline Fe phases.

Macrophyte Potential to Treat Leachate Contaminated with Wood Preservatives: Plant Tolerance and Bioaccumulation Capacity.

Pentachlorophenol and chromated copper arsenate (CCA) have been used worldwide as wood preservatives, but these compounds can toxify ecosystems when they leach into the soil and water. This study aimed to evaluate the capacity of four treatment wetland macrophytes, Phalaris arundinacea, Typha angustifolia, and two subspecies of Phragmites australis, to tolerate and treat leachates containing wood preservatives. The experiment was conducted using 96 plant pots in 12 tanks filled with three leachate concentrations compared to uncontaminated water. Biomass production and bioaccumulation were measured after 35 and 70 days of exposure. There were no significant effects of leachate contamination concentration on plant biomass for any species. No contaminants were detected in aboveground parts of the macrophytes, precluding their use for phytoextraction within the tested contamination levels. However, all species accumulated As and chlorinated phenols in belowground parts, and this accumulation was more prevalent under a more concentrated leachate. Up to 0.5 mg pentachlorophenol/kg (from 81 µg/L in the leachate) and 50 mg As/kg (from 330 µg/L in the leachate) were accumulated in the belowground biomass. Given their high productivity and tolerance to the contaminants, the tested macrophytes showed phytostabilization potential and could enhance the degradation of phenols from leachates contaminated with wood preservatives in treatment wetlands.

Keep and promote biodiversity at polluted sites under phytomanagement.

The phytomanagement concept combines a sustainable reduction of pollutant linkages at risk-assessed contaminated sites with the generation of both valuable biomass for the (bio)economy and ecosystem services. One of the potential benefits of phytomanagement is the possibility to increase biodiversity in polluted sites. However, the unique biodiversity present in some polluted sites can be severely impacted by the implementation of phytomanagement practices, even resulting in the local extinction of endemic ecotypes or species of great conservation value. Here, we highlight the importance of promoting measures to minimise the potential adverse impact of phytomanagement on biodiversity at polluted sites, as well as recommend practices to increase biodiversity at phytomanaged sites without compromising its effectiveness in terms of reduction of pollutant linkages and the generation of valuable biomass and ecosystem services.

Field evaluation of one Cu-resistant somaclonal variant and two clones of tobacco for copper phytoextraction at a wood preservation site.

A Cu-resistant somaclonal tobacco variant (NBCu 10-8-F1, C1), its BaG mother clone (C3), and the FoP tobacco clone (C2) were cultivated at a wood preservation site on Cu-contaminated soils (239-1290 mg Cu kg-1 soil range) and an uncontaminated control site (CTRL, 21 mg Cu kg-1) to assess their shoot DW yields and potential use for bioavailable Cu stripping. The Cu concentration in the soil pore water varied between 0.15 and 0.84 mg L-1. Influences of Cu exposure and soil treatments, i.e., untreated soil (Unt), soils amended with compost and either dolomitic limestone (OMDL) or zerovalent iron grit (OMZ), on plant growth and shoot ionome were determined. All transplants survived and grew even at high total soil Cu. Shoots were harvested after 3 months (cut 1). Subsequently, bottom suckers developed and were harvested after 2 months (cut 2). Total shoot DW yield (cuts 1 + 2) varied between 0.8 and 9.9 t DW ha-1 year-1 depending on tobacco cultivars, soil treatments, and soil Cu exposure. It peaked for all cultivars in the OMDL plots at moderate Cu exposure (239-518 mg kg-1 soil), notably for the C2 plants. Cut 2 contributed for 11-43% to total shoot DW yield. Increase in shoot DW yield diluted shoot Cu concentration. At low Cu exposure, total shoot Cu removal peaked for the variant. At moderate Cu exposure, shoot Cu concentrations were similar in all cultivars, but total shoot Cu removal was highest for the C2 plants. At high Cu exposure (753-1140 mg kg-1), shoot Cu concentrations peaked for the C2 plants in the Unt plots, the C1 and C2 plants in the OMZ plot, and the C3 ones in the OMDL plots. Shoot Cu removal (in g Cu ha-1 year-1) ranged from 15.4 (C2 on the CTRL soil) to 261.3 (C2 on moderately contaminated OMDL soils). The C2 plants phytoextracted more Cu than the C1 and C3 ones in the Unt plots and in the OMDL plots at moderate Cu exposure. In the OMDL plots with high Cu exposure, shoot Cu removal was highest for the C1 plants. Soil amendments improved shoot Cu removal through increase in either shoot DW yield (OMDL-3-fold) or shoot Cu concentration (OMZ-1.3-fold). Increased shoot Cu concentration induced an ionome imbalance with increased shoot Al, Fe, B, and Mg concentrations and decreased P and K ones. Copper concentrations in plant parts varied in decreasing order: roots > leaves > inflorescence (cymes including seeds) > stem, whereas Cu removal ranked as roots > stem = leaves > inflorescence.

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.

Functional activity and functional gene diversity of a Cu-contaminated soil remediated by aided phytostabilization using compost, dolomitic limestone and a mixed tree stand.

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.

Morphological and functional responses of a metal-tolerant sunflower mutant line to a copper-contaminated soil series.

The potential use of a metal-tolerant sunflower mutant line for biomonitoring Cu phytoavailability, Cu-induced soil phytotoxicity, and Cu phytoextraction was assessed on a Cu-contaminated soil series (13-1020 mg Cu kg-1) obtained by fading a sandy topsoil from a wood preservation site with a similar uncontaminated soil. Morphological and functional plant responses as well as shoot, leaf, and root ionomes were measured after a 1-month pot experiment. Hypocotyl length, shoot and root dry weight (DW) yields, and leaf area gradually decreased as soil Cu exposure rose. Their dose-response curves (DRC) plotted against indicators of Cu exposure were generally well fitted by sigmoidal curves. The half-maximal effective concentration (EC50) of morphological parameters ranged between 203 and 333 mg Cu kg-1 soil, corresponding to 290-430 µg Cu L-1 in the soil pore water, and 20 ± 5 mg Cu kg-1 DW in the shoots. The EC10 for shoot Cu concentration (13-15 mg Cu kg-1 DW) coincided to 166 mg Cu kg-1 soil. Total chlorophyll content and total antioxidant capacity (TAC) were early biomarkers (EC10: 23 and 51 mg Cu kg-1 soil). Their DRC displayed a biphasic response. Photosynthetic pigment contents, e.g., carotenoids, correlated with TAC. Ionome was changed in Cu-stressed roots, shoots, and leaves. Shoot Cu removal peaked roughly at 280 µg Cu L-1 in the soil pore water.

Tobacco, Sunflower and High Biomass SRC Clones Show Potential for Trace Metal Phytoextraction on a Moderately Contaminated Field Site in Belgium.

Phytoextraction could be a potential management option for diffusely Cd-Zn-Pb-polluted agricultural land in Northeast Belgium. The use of high yielding crops with a sufficiently high metal accumulation is preferred as these are expected to both gradually decontaminate the soil while generating an income through biomass valorization. To find out which high biomass crop possessed the highest and most constant (in time) phytoextraction potential on these soils, different plant species and different mutants or clones of each species, were evaluated during consecutive years. Biomass production and metal accumulation of pre-selected tobacco somaclonal variants (Nicotiana tabacum L.) and pre-selected sunflower mutants (Helianthus annuus L.) were investigated for two productivity years, while the phytoextraction potential of experimental poplar (Populus) and willow (Salix) in short rotation coppice (SRC) was assessed at the end of the second cutting cycle (after two times four growing seasons). The tobacco clones and the sunflower mutants showed efficient extraction of, respectively, Cd and Zn, while the highest simultaneous extractions of Cd and Zn were gained with some SRC clones. Variation in biomass production and metal accumulation were high for all crops over the years. The highest biomass production was observed for the experimental poplar clone of the crossing type Populus deltoides (P. maximowiczii x P. trichocarpa) with 9.9 ton DW per ha per year. The remediation period to reach legal threshold values for the pseudo-total content of Cd in this specific soil was estimated to be at least 60 years. Combining estimated phytoextraction potential and economic and environmental aspects, the SRC option is proposed as the most suitable crop for implementing metal phytoextraction in the investigated area.