Cu-contaminated Soil Research Articles

Biochar addition enhances remediation efficiency and rapeseed yield in copper-contaminated soil

IntroductionSoil contamination with copper (Cu) threatens ecological security and human health. Rapeseed demonstrates potential in remediating copper-contaminated soil, and biochar-assisted phytoremediation is increasingly being employed to improve remediation efficiency. However, the combined application of them has not been thoroughly studied in terms of the synergistic effects and the mechanisms of their interaction.MethodsIn this regard, this study conducted a pot experiment to evaluate biochar-assisted remediation under Cu-contaminated soil with varying biochar application rates; Furthermore, the plant physiological mechanism and soil physicochemical properties involved in the biocharrapeseed system was explored.ResultsOur results showed that the exchangeable pool of copper in soil decreased by 10.0% and 12.3% with adding 5% biochar (BC1) and 10% biochar (BC2) relative to control (BC0), respectively, prior to rapeseed cultivation. The rapeseed cultivation for one season further reclaimed 4.9%, 9.0%, and 13.6% of the available copper in this soil by root extraction under the BC0, BC1, and BC2 treatments, respectively. The overall copper concentration in plants decreased by 23.7% under BC2 and 13.3% under BC1 compared to BC0. However, the plant’s dry biomass at BC1 and BC2 treatments increased by 1.7-fold and 2.7-fold relative to BC0, which offset the negative impact of the decreased copper concentration on phytoremediation. Physiological analysis showed adding 10% biochar decreased the MDA content by 36% in the leaf and 49% in the root, compared to BC0. The transmission electron microscopy for cell wall ultrastructure in root tips showed that biochar addition in Cu-contaminated soil increased the mechanical strength of the celL wall, explicitly increasing the thickness of the secondary cell wall. Further cell wall components analysis revealed a remarkable increment of the pectin content in BC2 relative to BC0, increased by 56% in the leaf and 99% in the root, respectively. Additionally, 10% biochar application led to a roughly 2-fold increase in seed yield via ameliorating the soil physicochemical properties and increasing the rapeseed growth.DiscussionThese findings offer insights into synergistic rapeseed-biochar use for Cu-contaminated soil remediation.

Sustainable stabilization/solidification of Cu-contaminated seasonal frozen soil by epoxy resin: environmental risk assessment and engineering applicability

There are significantly toxic Cu contaminants in seasonal frozen soil areas under industrial production and mineral exploitation. However, the hydration reactions of mainstream alkaline curing agents such as cement are disturbed by the solidification and thawing of moisture, and their solidification/stabilization is ineffective to heavy metals. Therefore, there is an urgent requirement to optimize the use of current curing agents to improve the solidification/stabilization (S/S) effect of Cu contaminants in seasonal frozen soil areas. In this study, the Epoxy resin (EP) with excellent waterproofing and frost resistance was incorporated into artificially prepared Cu-contaminated soil to maintain a steady engineering application strength and inhibit the outward diffusion of toxic Cu contaminants under freeze-thaw cycles. The freeze-thaw resistance of EP-cured Cu-contaminated soil and the feasibility of EP remediation technology have been investigated, including mechanical properties, environmental effects and microstructure. The decline in mechanical strength and the increment in Cu leaching during freeze-thaw cycles are effectively suppressed by the remediation of EP. Even after 8 freeze-thaw cycles, there is merely a mechanical strength decline of 5%, solely a secondary Cu leaching of 4.74mg/L, and astonishingly a leachable index of 11.70 in the specimens with 12% EP dosage. The expansion phenomenon of pores and clay fractures under freeze-thaw cycles were gradually alleviated after incorporation of EP. The above results demonstrate the Cu-contaminated seasonal frozen soil remedied by EP are high-strength, basically non-toxic, and environmentally friendly material which are suitable for in-situ stabilization/solidification in seasonal frozen soil areas.

The effects of EDTA and Trichoderma species on growth and Cu uptake of maize (Zea mays) plants grown in a Cu-contaminated soil.

Metal contamination in soil poses a significant environmental concern worldwide, necessitating effective remediation strategies such as phytoremediation. The present study investigated the effects of EDTA dosage (1.5 and 3mmolkg-1) and two Trichoderma species (T. harzianum and T. aureoviride) on copper (Cu) content and growth of maize plants grown in a Cu-contaminated soil, as well as Cu fractionation in the soil. In the absence of EDTA, only inoculation with T. harzianum led to a significant increase in shoot biomass. Combining fungal inoculum with EDTA only yielded a significant increase in shoot biomass when using T. aureoviride at a low EDTA rate, highlighting the interplay between fungal species and EDTA rates on plant growth. Results also indicated that EDTA application increased Cu bioavailability, enhancing Cu dissolution and root (not shoot) Cu concentrations. Conversely, inoculation with both Trichoderma species reduced Cu mobility and bioavailability in soil, thereby decreasing the shoot Cu concentrations of plants. When combined with EDTA, only application of T. harzianum resulted in an enhanced shoot Cu concentration, whereas combined application of T. aureoviride and EDTA did not make a significant change compared to the corresponding control (no fungal inoculation, no EDTA), possibly due to a lower compatibility of the T. aureoviride isolate with EDTA. Our results demonstrated that EDTA application, in both non-inoculated and inoculated treatments, increased Cu availability by facilitating its redistribution and transformation from less plant-available fractions (residual, Fe/Mn oxide-bound, and carbonate-bound) to the more readily plant-available forms (water-soluble and exchangeable fractions). In conclusion, although individual Trichoderma application proved beneficial for phytostabilization by reducing Cu content and mitigating Cu toxicity in plants, the combined application of EDTA and a compatible Trichoderma isolate (here, the T. harzianum isolate) holds promise for enhancing the phytoextraction capacity of plants. Although using maize has the advantage of being a food crop, to optimize phytoextraction, plant species with superior metal tolerance and phytoextraction capabilities should be selected, exceeding those of maize.

Influence of soil pH and organic carbon content on the bioaccessibility of lead and copper in four spiked soils

Exploration of the association between heavy metal bioaccessibility (BAc) and soil properties is essential for rationalization of risk assessment and remediation of contaminated soil; however, the high complexity of soil systems often yield conflicting outcomes. To avoid erroneous conclusions, individual comparisons of soil properties is essential. Herein, we determined the changes in the BAc of Pb and Cu with the variation in soil pH and SOC content using Unified Bioaccessibility Research Group of Europe method, and validated these findings with in vivo mouse bioassays. Results indicated that the BAc of Pb and Cu in gastric and intestinal phases decreased by 1.76%–3.92% and 0.90%–3.27%, and by 0.41%–6.01% and 0.67%–1.59%, respectively, with every unit increase in soil pH. Furthermore, with every 1% increase in the absolute content of SOC, the BAc of Pb and Cu decreased by 4.04%–13.94% and 4.01%–34.7%, and by 8.98%–30.15% and 9.58%–20.03%, respectively. The in vivo bioassays results confirmed decrease in Pb concentrations in the liver, kidney, and blood of mice with the increase in Ferralosol pH and SOC content. These findings revealed that the health risks associated with accidental exposures to Pb- and Cu-contaminated soils with high pH and SOC level were relatively low, and the consistent in vivo and in vitro results for the BAc of Pb and Cu suggest the requirement for a swift and simple approach for assessing the risks of heavy metal contaminated soils. Thus, this study enhanced our understanding of the variations in risk assessments with soil properties of Pb- and Cu-contaminated soils, highlighting the role of soil characteristics in health risk assessment and remediation of contaminated soils.

Does copper contamination change thermotaxis of the soil arthropod Folsomia candida (Collembola)?

Behavioural thermoregulation (thermotaxis) is essential for soil invertebrates to evade thermal extremes in terrestrial environments. Extensive and continuous use of copper (Cu) based products has led to elevated Cu concentration in soils across the globe and in some areas reaching concentrations that are hazardous to soil invertebrates. We hypothesised that environmental stressors, for example, exposure to heavy metals may compromise the adaptive behavioural thermoregulation of organisms, but very little is known of such interactions. In this study, we chose Cu as a model toxicant and investigated the potential effect of Cu-contaminated soils on the behavioural thermoregulation of springtails (Folsomia candida). We measured the distribution of springtails when placed on a temperature gradient ranging from 6 to 46 °C and estimated their thermal preference as an indicator of behavioural thermoregulation. Results showed that within 60 min of being introduced to the thermal gradient, the distribution of springtails was unimodal with slight skewness towards high temperature. Springtails exhibited a consistent preferred temperature range of approximately 21–23 °C across all Cu exposure levels and time points. However, Cu contamination increased the frequency of springtails recorded along the gradient where temperature was above 30 °C. We interpreted this observation as Cu-exposed animals having an elevated risk of entering heat coma and not being able to evade noxious temperatures. We conclude that Cu contamination does not alter the thermal preference of F. candida but compromises their ability to tolerate extreme high temperature. Incorporating behavioural responses into ecotoxicological assessments provides ecologically relevant insights into the impacts of chemical pollution on soil ecosystems.

Molecular Mechanisms of Plant Responses to Copper: From Deficiency to Excess.

Copper (Cu) is an essential nutrient for plant growth and development. This metal serves as a constituent element or enzyme cofactor that participates in many biochemical pathways and plays a key role in photosynthesis, respiration, ethylene sensing, and antioxidant systems. The physiological significance of Cu uptake and compartmentalization in plants has been underestimated, despite the importance of Cu in cellular metabolic processes. As a micronutrient, Cu has low cellular requirements in plants. However, its bioavailability may be significantly reduced in alkaline or organic matter-rich soils. Cu deficiency is a severe and widespread nutritional disorder that affects plants. In contrast, excessive levels of available Cu in soil can inhibit plant photosynthesis and induce cellular oxidative stress. This can affect plant productivity and potentially pose serious health risks to humans via bioaccumulation in the food chain. Plants have evolved mechanisms to strictly regulate Cu uptake, transport, and cellular homeostasis during long-term environmental adaptation. This review provides a comprehensive overview of the diverse functions of Cu chelators, chaperones, and transporters involved in Cu homeostasis and their regulatory mechanisms in plant responses to varying Cu availability conditions. Finally, we identified that future research needs to enhance our understanding of the mechanisms regulating Cu deficiency or stress in plants. This will pave the way for improving the Cu utilization efficiency and/or Cu tolerance of crops grown in alkaline or Cu-contaminated soils.

Nighttime Warming Reduced Copper Concentration and Accumulation in Wheat Grown in Copper-Contaminated Soil by Affecting Physiological Traits

The changes in biomass (including yield), copper (Cu) concentration, and the accumulation of wheat (Triticum aestivum. L) in response to soil Cu pollution under nighttime warming had still not been explored. Hence, this study was carried out, and these variations were analyzed from a physiological perspective. Pot trials were performed at two levels of ambient temperatures (no-warming (NT) and average nighttime warming of 0.28 °C (WT)) and two levels of soil Cu concentrations (control check without Cu application (CK) and 100 mg/kg Cu application (Cu)). Soil was collected from the carbonate cinnamon soil region of central China. The warming effects of the passive nighttime warming system were prominent, and the average increment was 0.28 °C. Antioxidant enzyme activities were promoted by warming (p < 0.05) and Cu. The highest yield was achieved in NT-Cu, mainly attributed to relatively strong root activity and photosynthesis caused by supplemental Cu, but the Cu concentration in its grains was close to the threshold (10 mg/kg) for Cu concentration in foodstuff and could present a potential food safety risk. Though nighttime warming did not increase the total biomass and yield of wheat, it decreased the Cu accumulation of wheat grown in Cu-contaminated soil, especially in grains. Moreover, WT-CK and WT-Cu increased the Cu concentration in the roots and glumes and reduced the Cu concentration in grains by 13.09% and 55.84%, respectively, probably because of a lower transpiration rate. Among them, the Cu concentration of grains in WT-Cu was the lowest and significantly lower than other applications. Our findings reveal that nighttime warming has the potential to reduce the Cu risk of grains in wheat grown in the Cu-contaminated carbonate cinnamon soil region of central China and could then provide a theoretical reference for risk assessment of food quality for wheat subjected to dual stress from nighttime warming and Cu pollution.

Mitigating potential of polystyrene microplastics on bioavailability, uptake, and toxicity of copper in maize (Zea mays L.)

The coexistence of polystyrene microplastics (PSMPs) and copper (Cu) has become a pressing issue for croplands. However, limited literature is available regarding the interaction of PSMPs with essential micronutrients in Cu-contaminated soils. Therefore, the present study aimed to analyze the immobilization potential of PSMPs for micronutrient bioavailability in soil and Cu toxicity in maize (Zea mays L.). A pot experiment was conducted with maize variety “Islamabad gold” exposed to varying Cu concentrations (0, 50, 100, 200, and 400 mg/kg) and PSMPs (150–250 μm size, 0, 1, and 3% w/w) via soil spiking for 60 days. The concentrations of essential micronutrients (Zn, Cu, Mn, Fe) in soil and plant tissues were measured using an atomic absorption spectrophotometer. Moreover, malondialdehyde (MDA) and antioxidant activities (superoxide dismutase, ascorbate peroxidase, catalase, and peroxidase) were recorded. The concentration of Cu showed significant reduction in post-harvesting soil by 21, 24.8, 27.6, 29.2, and 30.2% from Cu0 to Cu400 mg/kg respectively from pre-sowing soil. On the other hand, the addition of 1%PSMPs and 3%PSMPs declined Cu by 16, 21.6, 24.4, 25.9, 27.8, and 12.6, 16.5, 19.9, 23.2, 25% from Cu0 to Cu400 mg/kg respectively. Maize showed significant improvement in growth under combined exposure of Cu and 3% PSMPs compared to individual exposure. The MDA level was decreased under the combined presence of Cu and PSMPs compared to individual Cu exposure. The percentage difference with 1%PSMPs was 98.1, 95.0, 92.0, 90.0, and 89.6%, while with 3%PSMPs was 93.2, 93.2, 87.7, 81.4, and 79.2% from Cu0 to Cu400 mg/kg respectively. Moreover, the impact of PSMPs was more prominent at a 3% dose compared to a 1% dose. The findings provided significant knowledge about the potential of PSMPs to mitigate Cu toxicity in maize. Future research should incorporate a variety of particle size distributions at natural conditions for variety-specific differences.

Unravelling the combined impacts of drought and Cu in barley plants – double trouble?

The occurrence of drought in soils, particularly in those contaminated by metals, poses a current threat to crops, as these factors can interact and induce unique stress responses. Therefore, this study mainly focused on understanding the crosstalk between drought and copper (Cu) stress in the physiology of the barley (Hordeum vulgare L.) plant. Using a bifactorial experimental design, seedlings were grown in a natural soil under the following treatments: plants continuously irrigated in uncontaminated soil for 14 days (control); plants continuously irrigated in Cu-contaminated soil (115 mg Cu kg−1) for 14 days (Cu); plants only irrigated during the initials 7 days of growth in uncontaminated soil (drought); plants co-exposed to Cu and drought (combined). After 14 days of growth, the results revealed that drought prevented Cu bioaccumulation in barley roots, which were still severely affected by the metal, both individually and in combination with the water deficit. Furthermore, individual and combined exposure to these stressors resulted in impaired photosynthetic performance in barley plants. Despite the increased activation of enzymatic and non-enzymatic antioxidant defence mechanisms, particularly in the green organs, the plants co-exposed to both stress factors still showed higher oxidative damage, severely impacting biomass production.

Phytoremediation & Valorization of Cu-contaminated Soils Through Cannabis sativa (L.) Cultivation: A Smart Way to Produce Cannabidiol (CBD) in Mediterranean Soils

PurposeThe potential of industrial hemp (Cannabis sativa L.) for phytoremediation of copper-contaminated Mediterranean soils was investigated. The accumulation of copper in hemp’s parts and the effect of contaminated soil on the production of cannabidiol (CBD) were, also, of primary concern.MethodsTwo soil types, a Clay Loam & a Sandy Clay Loam, were exposed to two levels of Cu contamination, with low & high Cu concentrations. The soils’ total and available Cu content, along with Cu in plant tissues, were determined. Height, fresh aboveground biomass and leaf chlorophyll content were further evaluated. Furthermore, the amount of cannabidiol (CBD) in the hemp flowers was determined.ResultsHemp appeared to be highly resistant, as can successfully grow in both soil types having low or high Cu levels. The maximum Cu content was detected in the highest part of the plant roots, with a progressive decline towards the upper parts of the plant (CuRoots > CuShoots > CuLeaf), in both soil types. It has been observed that more than 50% of the plant’s copper concentration is accumulated in the roots. Furthermore, the hemp plants cultivated in high Cu-contaminated soils produced greater amounts of cannabidiol (CBD).ConclusionΗemp proved to be a promising plant for phytostabilization in Cu-contaminated soils, as its above-ground biomass is almost free of metals and can be used further for fiber production. The presence of Cu in soils did not appear to disrupt the production of the important secondary metabolite CBD, but rather increased following increasing soil Cu content.Graphical

Potential of herbaceous plant species for copper (Cu) accumulation.

The removal of copper (Cu) in soils by green technology is less treated with urgency, as it is a plant micronutrient. We examined the efficiency of Cu shoot accumulation by herbaceous plants in Cu-contaminated and non-contaminated soils in Trhové Dusniky and Podles, respectively, in the Czech Republic. The total soil Cu content of 81mgkg-1 in Trhové Dusniky indicated a slight contamination level compared to 50mgkg-1, the permissible value by WHO, and < 35 in Podlesí, representing a clean environment. The Cu content was above the permissible value in plants (10mgkg-1 by WHO) in herbaceous speciesat the control site without trees: Stachys palustris L. (10.8mgkg-1), Cirsium arvense L. (11.3mgkg-1), Achillea millefolium L. (12.1mgkg-1), Anthemis arvense L. (13.2mgkg-1), and Calamagrostis epigejos L. (13.7mgkg-1). In addition, Hypericum maculatum Crantz (10.6mgkg-1), Campanula patula L. (11.3mgkg-1), C. arvense (15mgkg-1), and the highest accumulation in shoot of Equisetum arvense L. (37.1mgkg-1), all under the canopy of trees at the uncontaminated site, were above the WHO value. Leucanthemum Vulgare (Lam.) and Plantago lanceolata L. recorded 11.2mgkg-1 and 11.5mgkg-1, respectively, in the soil of the Cu-contaminated site. These herbaceous species can support the phyto-management of Cu-contaminated soils, especially E. arvense. Critical attention is well-required in the medicinal application of herbaceous plants in treating human ailments due to their Cu accumulation potentials above the threshold. Spontaneous surveys and analysis of Cu speciation in herbaceous species can reveal suitable plants to decontaminate soils and provide caution on consumable products, especially bioactive compounds.

Epoxy resin for solidification/stabilization of soil contaminated with copper (II): Leaching, mechanical, and microstructural characterization

Among different types of heavy metal-contaminated soil, copper (Cu)-contaminated soil is very serious, and the Cu concentration in it is usually very high. It is common to solidify/stabilize Cu-contaminated soil using alkaline cementitious material. However, the remediated Cu-contaminated soil fails to meet the requirements of environmental safety and load-bearing capacity. This dilemma in the remediation of Cu-contaminated soil hinders the effective utilization of land resources. In this study, epoxy resin (EP) was utilized to solidify/stabilize Cu-contaminated soil due to its stable and rapid curing performance and excellent resistance to acid, alkali, and salt erosion. The mechanical properties, environmental effects, and curing mechanism of EP-cured Cu-contaminated soil were investigated. The results showed that the application of EP significantly enhanced the unconfined compressive strength (UCS), cohesion and internal friction angle of Cu-contaminated soil. All specimens met the UCS criterion specified by the United States Environmental Protection Agency (USPEA), namely no less than 0.35 MPa, which indicated that those EP-cured Cu-contaminated soil were qualified for practical engineering applications. According to the toxicity characteristic leaching procedure (TCLP), the application of EP enhanced the stability of Cu in Cu-contaminated soil. The leaching index of Cu ranged from 11 to 14. A high leaching index showed that the S/S treatment was safe and effective and the remediated Cu-contaminated soil satisfied the environmental requirement for heavy metals. This study confirmed the feasibility of utilizing EP in the solidification/stabilization (S/S) technology to convert high-concentration Cu-contaminated soil into secure and stable engineering materials. The remediation of Cu-contaminated soil by EP lays a solid foundation for the safe treatment and reuse of heavy metal-contaminated land resources.

Copper bioreduction and nanoparticle synthesis by an enrichment culture from a former copper mine.

Microorganisms can facilitate the reduction of Cu2+ , altering its speciation and mobility in environmental systems and producing Cu-based nanoparticles with useful catalytic properties. However, only a few model organisms have been studied in relation to Cu2+ bioreduction and little work has been carried out on microbes from Cu-contaminated environments. This study aimed to enrich for Cu-resistant microbes from a Cu-contaminated soil and explore their potential to facilitate Cu2+ reduction and biomineralisation from solution. We show that an enrichment grown in a Cu-amended medium, dominated by species closely related to Geothrix fermentans, Azospira restricta and Cellulomonas oligotrophica, can reduce Cu2+ with subsequent precipitation of Cu nanoparticles. Characterisation of the nanoparticles with (scanning) transmission electron microscopy, energy-dispersive x-ray spectroscopy and electron energy loss spectroscopy supports the presence of both metallic Cu(0) and S-rich Cu(I) nanoparticles. This study provides new insights into the diversity of microorganisms capable of facilitating copper reduction and highlights the potential for the formation of distinct nanoparticle phases resulting from bioreduction or biomineralisation reactions. The implications of these findings for the biogeochemical cycling of copper and the potential biotechnological synthesis of commercially useful copper nanoparticles are discussed.

Farmyard manure, a potential organic additive to reclaim copper and Macrophomina phaseolina stress responses in mash bean plants

In the era of global warming, stress combinations instead of individual stress are realistic threats faced by plants that can alter or trigger a wide range of plant responses. In the current study, the cumulative effect of charcoal rot disease caused by notorious fungal pathogen viz., Macrophomina phaseolina was investigated under toxic levels of copper (Cu) in mash bean, and farmyard manure (FYM) was employed to manage stress. Therefore, Cu-spiked soil (50 and 100 mg/kg) was inoculated with the pathogen, and amended with 2% FYM, to assess the effect of intricate interactions on mash bean plants through pot experiments. Results demonstrated that the individual stress of the pathogen or Cu was more severe for morpho-growth, physio-biochemical, and expression profiles of stress-related genes and total protein in mash bean plants as compared to stress combinations. Under single Cu stress, a significant amount of Cu accumulated in plant tissues, particularly in roots than in upper ground tissues, while, under stress combination less Cu accumulated in the plants. Nonetheless, 2% FYM in soil encountered the negative effect of stress responses provoked by the pathogen, Cu, or both by improving health markers (photosynthetic pigments, reducing sugar, total phenolics) and oxidative stress markers (catalase, peroxidase, and polyphenol oxidase), together with regulating the expression of stress-related genes (catalase, ascorbate peroxidase, and cytokinin-resistant genes), and proteins, besides decreasing Cu uptake in the plants. FYM worked better at lower concentrations (50 mg/kg) of Cu than at higher ones (100 mg/kg), hence could be used as a suitable option for better growth, yield, and crop performance under charcoal rot disease stress in Cu-contaminated soils.

Tolerance and Environmental Effects of Chinese Milk Vetch (Astragalus sinicus L.) Planted in Cu-Contaminated Soil

Tolerance and Environmental Effects of Chinese Milk Vetch (Astragalus sinicus L.) Planted in Cu-Contaminated Soil

Toxicity, physiological, and morphological alterations of Indian camphorweed (Pluchea indica) in response to excess copper.

Indian camphorweed (Pluchea indica (L.) Less.) is used as herbal tea due to the presence of volatile aromatic oils and several phytochemical compounds. The aim of this study was to assess the impact of copper (Cu) contamination on the physiology and morphology of P. indica, and the health risks associated with its consumption as tea. The cuttings of P. indica were subjected to 0mM (control), 5mM (low Cu), and 20mM (excess Cu) of CuSO4 treatments for 1, 2, and 4weeks. Thereafter, Cu contamination as well as physiological and morphological parameters were assessed. Cu accumulation was higher in the root tissues of plants (25.8 folds higher as compared to the leaves) grown under 20mM CuSO4 for 4weeks. This increased Cu accumulation resulted in the inhibition of root length, root fresh weight, and root dry weight. Cu concentration was found maximum (1.36μgg-1 DW) in the leaf tissues under 20mM Cu exposure for 4weeks, with the highest target hazard quotient (THQ = 1.85), whereas Cu was not detected in control. Under exposure to 20mM Cu treatment for 4weeks, leaf greenness, maximum quantum yield of photosystem II, and photon yield of photosystem II diminished by 21.4%, 16.1%, and 22.4%, respectively, as compared to the control. Leaf temperature was increased by 2.5°C, and the crop stress index (CSI) exceeded 0.6 when exposed to 20mM Cu treatment for 2 and 4weeks; however, the control had a CSI below 0.5. This led to a reduced transpiration rate and stomatal conductance. In addition, the net photosynthetic rate was also found sensitive to Cu treatment, which resulted in decreased shoot and root growth. Based on the key results, it can be suggested that P. indica herbal tea derived from the foliage of plants grown under a 5mM Cu level (0.75μgg-1 DW) with a target hazard quotient below one aligns with the recommended dietary intake of Cu in leafy vegetables. The study recommends choosing cuttings from plants with a small canopy as plant material in the greenhouse microclimates to validate the growth performance in the Cu-contaminated soil and simulate the natural shrub architecture and life cycle.

Phytoremediation of copper-contaminated soils by drip or sprinkling irrigation coupled with intercropping.

Intercropping improves the phytoremediation of soil trace metal contamination. Dripping irrigation could further promote the phytoremediation of trace metals by influencing their speciation and total amount in soil. However, there is currently insufficient information elucidating this synergistic effect. In this study, the combined effect of drip irrigation and intercropping on the phytoremediation of Cu-contaminated soil was testified by investigating the changes of Cu spatial distribution and speciation in soil irrigated by dripping or sprinkling methods, as well as Cu bioconcentration factor and translocation factor by plants. Results showed that after a 30-day drip irrigation, the Cu level in soils near the drip outlet decreased by 4.7% and that in Triticum aestivum L. (T. aestivum) roots intercropped with Helianthus annuus L. (H. annuus) and Zea mays L. (Z. mays) dropped by 53.2% and 25.1%, respectively, relative to sprinkler irrigation. Meanwhile, the total Cu and exchangeable Cu levels in soils 6cm away from the drip outlet increased by 10.8% and 20.4% after 30days of drip irrigation, leading to 41.1% and 40.0% increases of Cu content in remediation plants H. annuus and Z. mays seedlings as compared to the values by sprinkler irrigation. Therefore, the drip irrigation enhanced the effect of intercropping on Cu phytoremediation.

Changes in morphology and yield attributes of Nigella sativa L. by copper (Cu) induced toxicity

The present study was conducted to evaluate the impact of Cu-induced toxicity on the growth, yield parameters, and Cu accumulation in the seeds of black cumin (Nigella sativa L.) plants. In this experiment, plants grown in earthen pots filled with 3 kg of garden soil were contaminated with different dilutions (1.5, 2.0, 2.5, 3.0, 3.5, 4.0) of Cu in millimoles (mM), except for the control. Growth, yield, and Cu bioaccumulation parameters were recorded and analyzed at the harvesting stage. Results revealed that all doses of Cu show a considerable negative effect on the growth and yield parameters of black cumin as the concentration of Cu increased in the soil and reduced all of the growth parameters significantly (p ≤ 0.05). With each Cu dose, the levels of chlorophyll a and b in the tissues of fresh leaves decreased significantly (p ≤ 0.05). Fruit and seed yield also decreased significantly (p ≤ 0.05) from the lower to the higher dose of Cu (1.5 mM to 4.0 mM). Cu bioaccumulation levels in the seeds of all treated plants (Control to 4.0 mM Cu) are under the threshold limit for Cu set by the Food Safety and Standards Authority of India (FSSAI) and World Health Organization (WHO) for spices and are safe for human consumption. Finally, due to the low seed yield in Cu-contaminated soil, it is advised to farmers that Nigella sativa should not be grown in Cu-contaminated soil to avoid economic loss.

Organic fertilization and mycorrhization increase copper phytoremediation by Canavalia ensiformis in a sandy soil

Organic fertilization and mycorrhization can increase the phytoremediation of copper-contaminated soils. The time of vermicomposting alters the properties of vermicompost, which can affect copper's availability and uptake. Therefore, this study sought to evaluate the effect of different organic fertilizers and mycorrhization on copper-contaminated soil phytoremediation. The soil was contaminated with 100 mg Cu kg-1 dry soil and received mineral fertilizer (MIN), bovine manure (CM), and vermicompost produced in 45 days (V45) or 120 days (V120), all in doses equivalent to 40 mg kg-1 dry soil of phosphorus. Half of the jack bean (Canavalia ensiformis) plants were inoculated with the arbuscular mycorrhizal fungus Rhizophagus clarus. At plant flowering, the dry mass and concentrations of Cu, Zn, Mn, Ca, Mg, P, and K in the soil, solution, and plant tissue were determined, in addition to mycorrhizal colonization, nodulation, photosynthetic pigments, and oxidative stress enzyme activity. Organic fertilization increased plant growth and copper accumulation in aerial tissues. These effects were more evident with the V120, making it suitable for use in copper phytoextraction. Mycorrhization increased root and nodule dry mass, making it recommended for phytostabilization. C. ensiformis nodulation in Cu-contaminated soils depends on vermicompost fertilization and mycorrhization. Hence, the copper phytoremediation by C. ensiformis is increased by using organic fertilization and mycorrhization.

Mechanism for combined application of biochar and Bacillus cereus to reduce antibiotic resistance genes in copper contaminated soil and lettuce

The remediation of agricultural soil contaminated by antibiotic resistance genes (ARGs) is of great significance for protecting food safety and human health. Reducing the availability of copper in soil may control coresistance to ARGs. However, the feasibility of applying nano-biochar and Bacillus cereus to mitigate the spread of ARGs in Cu contaminated soil remains unclear. Therefore, this study investigated the use of biochar with different particle sizes (2 % apple branch biochar and 0.5 % nano-biochar) and 3 g m−2B. cereus in a 60-day pot experiment with growing lettuce. The effects of single and combined application on the abundances of ARGs in Cu-contaminated soil (Cu = 200 mg kg−1) were compared, and the related mechanisms were explored. Studies have shown that the addition of biochar alone is detrimental to mitigating ARGs in soil–lettuce systems. The combined application of 3 g m−2B. cereus and 0.5 % nano-biochar effectively inhibited the proliferation of ARGs in Cu-contaminated soil, and 3 g m−2B. cereus effectively inhibited the proliferation of ARGs in lettuce. Partial least squares-path modeling and network analysis showed that bacterial communities and mobile genetic elements were the key factors that affected the abundances of ARGs in rhizosphere soil, and Cu resistance genes and bioavailable copper (acid extractable state Cu (F1) + reducing state Cu (F2)) had less direct impacts. The bacterial community was the key factor that affected the abundances of ARGs in lettuce. Rhodobacter (Proteobacteria), Corynebacterium (Actinobacteria), and Methylobacterium (Proteobacteria) may have been hosts of ARGs in lettuce plants. B. cereus and nano-biochar affected the abundances of ARGs by improving the soil properties and reducing the soil bioavailability of Cu, as well as directly or indirectly changing the bacterial community composition in soil and lettuce, thereby impeding the transport of ARGs to aboveground plant parts.