Correspondance auteur: P. Hinsinger e-mail: philippe.hinsinger@supagro.inra.fr | International audience | We evaluated how root-induced changes in rhizosphere pH varied and interacted with Cu availability and bioavailability in an acidic soil. Rape was grown on a Cu-contaminated acidic soil, which had been limed at 10 rates. Soil Cu bioavailability was not influenced by liming. However, liming significantly decreased CaCl2-extracted Cu for pH between 3.7 and 5.1. Little effect was found for pH above 5.1. For soil pH < 4.4, CaCl2-Cu contents were smaller in rhizosphere than uncropped soil. Rhizosphere alkalisation occurred at pH < 4.8, while acidification occurred at greater pH. This explained the changes of CaCl2-Cu in the rhizosphere at low pH and the absence of pH dependency of Cu bioavailability to rape. In addition, apoplastic Cu in roots increased with increasing soil pH, most probably as a result of increased dissociation and affinity of cell wall compounds for Cu

Melatonin (M) is a pleiotropic molecule that improves plant growth and increases heavy metal tolerance. The role of M for improving plant growth and tolerance under cadmium (Cd) stress, and mitigation of Cd-induced toxicity has not yet been sufficiently examined. Therefore, here we conducted a glasshouse experiment to explore the influence of various M dosages on Cd detoxification and stress-tolerance responses of Brassica napus under high Cd content (30 mg kg⁻¹). The effects of M on the modulation of Cd tolerance in B. napus plants have been investigated using various growth attributes, Cd accumulation and tolerance indices, and secondary metabolic parameters. We found that Cd stress inhibited root growth (by 11.9%) as well as triggered reactive oxygen species accumulation (by 31.2%) and MDA levels (by 18.7%); however, exogenous M substantially alleviated the adverse effect of oxidative stress by decreasing levels of H₂O₂ (by 38.7%), MDA (by 13.8%) and EL (by 1.8%) in the Cd-stressed plants, as compared to the M-untreated plants (control). Interestingly, exogenous M reduced Cd accumulation in roots (∼48.2–58.3-fold), stem (∼2.9–5.0-fold) and leaves (∼4.7–6.6-fold) compared to control plants, which might be due to an M-induced defense and/or detoxification response involving a battery of antioxidants. Overall, addition of the exogenous M to the Cd-stressed plants profoundly enhanced Cd tolerance in B. napus relative to control plants. These results suggested the biostimulatory role (at the physiological and molecular level) of M in improving growth, Cd tolerance, and Cd detoxification in B. napus, which indicate the potentiality of M for green remediation of Cd contaminated soils. This green trial would provide a reference for producing renewable bioenergy crops under Cd stress in contaminated soils. However, these recommendations should be verified under field conditions and the potential mechanisms for the interaction between Cd and M should be explicitly explored.

A large amounts of arable land is facing a high risk of hexavalent chromium (Cr(VI)) pollution, which requires remediation using a low toxic agent. In this study, the remediation effect of amorphous iron pyrite (FeS₂₍ₐₘ₎) on Cr(VI) in Cr(VI)-contaminated soil was evaluated by systematically analyzing the variation of the leachability, bioaccessibility, phytotoxicity, and long-term stability of the remediated soil. The effectiveness of FeS₂₍ₐₘ₎ on the leachability was assessed by alkaline digestion and the toxicity characteristic leaching procedure (TCLP); the effect on the bioaccessibility was evaluated via the physiologically based extraction test (PBET) and the Tessier sequential extraction; the effect on the phytotoxicity was assessed via phytotoxicity bioassay (seed germination experiments) based on rape (Brassica napus L.) and cucumber (Cucumis Sativus L.), and the long-term stability of the Cr(VI)-remediated soil was appraised using column tests with groundwater and acid rain as the influents. The results show that FeS₂₍ₐₘ₎, with a stoichiometry of 4× exhibited a high efficiency in the remediation of Cr(VI) and decreased its leachability and bioaccessibility during the 30-day remediation period. In addition, seed germination rate, accumulation and translocation of Cr, and root and shoot elongation of rape and cucumber of remediated soil are not significantly different from those of clean soil, illustrating that FeS₂₍ₐₘ₎ is suitable for remediating Cr(VI) contaminated arable soil. The stabilization of Cr(VI) in contaminated soil using FeS₂₍ₐₘ₎ was maintained for 1575 days. The long-term effectiveness was further confirmed by the increasing amount of free Fe and Mn in the effluent and the decreasing redox potential. In summary, FeS₂₍ₐₘ₎ has an excellent efficiency for the remediation of Cr(VI), demonstrating it is a very promising alternative for use in the contaminated arable soil.

Sclerotinia stem rot (SSR), a soil-borne plant disease, cause the yield loss of oilseed rape. Selenium (Se), a beneficial element of plant, improves plant resistance to pathogens, and regulates microbial communities in soil. Soil microbial communities has been identified to play an important role in plant health. We studied whether the changes in soil microbiome under influence of Se associated with oilseed rape health. SSR disease incidence of oilseed rape and soil biochemical properties were investigated in Enshi district, “The World Capital of Selenium”, and soil bacterial and fungal communities were analyzed by 16S rRNA and ITS sequencing, respectively. Results showed that Se had a strong effect on SSR incidence, and disease incidence inversely related with plant Se concentration. Besides, soil Se enhanced the microbiome diversities and the relative abundance of PGPR (plant growth promoting rhizobacteria), such as Bryobacter, Nitrospirae, Rhizobiales, Xanthobacteraceae, Nitrosomonadaceae and Basidiomycota. Furthermore, Soil Se decreased the relative abundance of pathogenic fungi, such as Olpidium, Armillaria, Coniosporium, Microbotryomycetes and Chytridiomycetes. Additionally, Se increased nitrogen metabolism, carbohydrate metabolism and cell processes related functional profiles in soil. The enrichment of Se in plants and improvement of soil microbial community were related to increased plant resistance to pathogen infection. These findings suggested that Se has potential to be developed as an ecological fungicide for biological control of SSR.

Heavy metals contamination in agricultural soil has become a worldwide problem, and soil characteristics modulate metal availability in soils. Four field experiments were conducted simultaneously to evaluate concentration and distribution of cadmium (Cd) and lead (Pb) in 39 oilseed rape cultivars at four agricultural locations with different contamination levels of Cd and Pb, as well as the influence of soil characteristics together with soil total and bioavailable Cd and Pb concentration on metal transfer from soil to oilseed rape. Shoot concentrations of Cd and Pb in oilseed rape cultivars ranged from 0.09 to 3.18 and from 0.01 to 10.5 mg kg⁻¹ across four sites. For most cultivars, Cd concentration in root or shoot were higher than pod and lowest in seed, while the highest Pb concentration was observed in root followed by shoot and seed. Stepwise multiple linear regression analysis allows for a better estimation of Cd and Pb concentration in oilseed rape while taking soil properties into consideration. The results demonstrated that Cd and Pb concentration in oilseed rape were correlated with soil organic matter (OM), cation exchange capacity (CEC), available phosphorus (AP), available potassium (AK), sand, soil total and available Cd and Pb concentration, and R² varied from 0.993 to 0.999 (P < 0.05). The Cd and Pb levels found in oilseed rape indicated its phytoextraction potential for Cd and Pb co-contaminated agricultural soils in winter without stopping agricultural activities.

Metabolic profiling in plants can be used to differentiate between treatments and to search for biomarkers for exposure. A methodology for processing Ultra-High-Performance Liquid Chromatography–Diode-Array-Detection data is devised. This methodology includes a scheme for selecting informative wavelengths, baseline removal, retention time alignment, selection of relevant retention times, and principal component analysis (PCA). Plant crude extracts from rapeseed seedling exposed to sublethal concentrations of glyphosate are used as a study case. Through this approach, plants exposed to concentrations down to 5 μM could be distinguished from the controls. The compounds responsible for this differentiation were partially identified and were different from those specific for high exposure samples, which suggests that two different responses to glyphosate are elicited in rapeseed depending on the level of exposure. The PCA loadings indicate that a combination of other metabolites could be more sensitive than the response of shikimate to detect glyphosate exposure.

Most available exposure–response relationships for assessing crop loss due to elevated ozone (O3) have been established using data from chamber and open-top chamber experiments, using a simulated constant O3 concentration exposure (square wave), which is not consistent with the diurnal variation of O3 concentration that occurs in nature. We investigated the response of oilseed rape (Brassica napus L.) to O3 as affected by two exposure regimes: one with a diurnal variation (CF100D) and another with a constant concentration (CF100). Although the two exposure regimes have the same mean O3 concentration and accumulated O3 concentration above 40 ppb (AOT40), our results show that O3 at CF100D reduced biomass and number of pods/plant more than O3 at CF100. Both O3 exposures resulted in larger seed weights/100 pods compared to CF. Numbers of seeds/100 pods were reduced by CF100, while numbers of seeds/100 pods in the CF100D chambers were comparable to those in CF. Our results suggest that chamber experiments that use a constant O3 exposure may underestimate O3 effects on biomass and yields. Diurnal variation of O3 concentration should be considered when designing O3 exposure experiment.

The growth of transgenic canola (Brassica napus) expressing a gene for the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase was compared to non-transformed canola exposed to flooding and elevated soil Ni concentration, in situ. In addition, the ability of the plant growth-promoting bacterium Pseudomonas putida UW4, which also expresses ACC deaminase, to facilitate the growth of non-transformed and transgenic canola under the above mentioned conditions was examined. Transgenic canola and/or canola treated with P. putida UW4 had greater shoot biomass compared to non-transformed canola under low flood-stress conditions. Under high flood-stress conditions, shoot biomass was reduced and Ni accumulation was increased in all instances relative to low flood-stress conditions. This is the first field study to document the increase in plant tolerance utilizing transgenic plants and plant growth-promoting bacteria exposed to multiple stressors. Using transgenic plants and plant growth-promoting bacteria as phytoremediation methods increased plant tolerance at a metal-contaminated field site under low flood conditions.

Soil chemistry of toxic metalloids and metals differs, making their simultaneous removal difficult. Soil contaminated with As, Pb, Zn and Cd was washed with oxalic acid, Na-dithionite and EDTA solution. Toxic elements were removed from the washing solution by alkalinisation with CaO to a pH 12.5: As was co-precipitated with Fe from Fe-EDTA chelate formed after the soil washing. The toxic metals precipitated after substitution of their EDTA chelates with Ca. The novel method was scaled up on the ReSoil® platform. On average, 60, 76, 29, and 53% of As, Pb, Zn, and Cd were removed, no wastewater was generated and EDTA was recycled. Addition of zero-valent iron reduced the toxic elements’ leachability. Remediation was most effective for As: phytoaccessibility (CaCl₂ extraction), mobility (NH₄NO₃), and accessibility from human gastric and gastrointestinal phases were reduced 22, 104, 6, and 51 times, respectively. Remediation increased pH but had no effect on soil functioning assessed by fluorescein diacetate hydrolysis, dehydrogenase, β-glucosidase, urease, acid and alkaline phosphatase activities. Brassica napus produced 1.9 times more biomass on remediated soil, accumulated no As and 5.0, 2.6, and 9.0 times less Pb, Zn and Cd, respectively. We demonstrated the novel remediation technology as cost-efficient (material cost = 41.86 € t⁻¹) and sustainable.