Conjoint analysis of physio-biochemical, transcriptomic, and metabolomic reveals the response characteristics of solanum nigrum L. to cadmium stress.

Cadmium (Cd) is a nonessential element in plants and has adverse effects on the growth and development of plants. However, the molecular mechanisms of Cd phytotoxicity, tolerance and accumulation in hyperaccumulators Solanum nigrum L. has not been well understood. Here, physiology, transcriptome, and metabolome analyses were conducted to investigate the influence on the S. nigrum under 0, 25, 50, 75 and 100 µM Cd concentrations for 7 days. Pot experiments demonstrated that compared with the control, Cd treatment significantly inhibited the biomass, promoted the Cd accumulation and translocation, and disturbed the balance of mineral nutrient metabolism in S. nigrum, particularly at 100 µM Cd level. Moreover, the photosynthetic pigments contents were severely decreased, while the content of total protein, proline, malondialdehyde (MDA), H2O2, and antioxidant enzyme activities generally increased first and then slightly declined with increasing Cd concentrations, in both leaves and roots. Furthermore, combined with the previous transcriptomic data, numerous crucial coding-genes related to mineral nutrients and Cd ion transport, and the antioxidant enzymes biosynthesis were identified, and their expression pattern was regulated under different Cd stress. Simultaneously, metabolomic analyses revealed that Cd treatment significantly changed the expression level of many metabolites related to amino acid, lipid, carbohydrate, and nucleotide metabolism. Metabolic pathway analysis also showed that S. nigrum roots activated some differentially expressed metabolites (DEMs) involved in energy metabolism, which may enhance the energy supply for detoxification. Importantly, central common metabolism pathways of DEGs and DEMs, including the "TCA cycle", "glutathione metabolic pathway" and "glyoxylate and dicarboxylate metabolism" were screened using conjoint transcriptomics and metabolomics analysis. Our results provide some novel evidences on the physiological and molecular mechanisms of Cd tolerance in hyperaccumulator S. nigrum plants.

Combined effects of cadmium and sulfamethoxazole on Eisenia fetida: Insights into accumulation, subcellular partitioning, biomarkers and toxicological responses.

Cadmium (Cd) and sulfamethoxazole (SMX) frequently coexist in farmlands, yet their synergistic toxicological impacts on terrestrial invertebrates remain unexplored. In this study, earthworms were exposed to artificial soils percolated with Cd (5 mg/kg), SMX (5 mg/kg) or combination of them for 7 days, followed by a 12-day elimination phase in uncontaminated soil. The uptake of Cd and SMX by the earthworms, along with their subcellular distribution, was meticulously analyzed. Additionally, a suite of biomarkers-including superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and weight loss-were evaluated to assess the health status of the earthworms and the toxicological effects of the Cd and SMX mixture. Notably, the cotreatment with Cd and SMX resulted in a significantly higher weight loss in Eisenia fetida (41.25 %) compared to exposure to Cd alone (26.84 %). Moreover, the cotreatment group exhibited substantially higher concentrations of Cd in the total internal body, fraction C (cytosol), and fraction E (tissue fragments and cell membranes) in Eisenia fetida compared to Cd alone counterparts. The combined exposure also significantly elevated the SMX levels in the total body and fraction C compared with the SMX-only treated earthworms. Additionally, Eisenia fetida subjected to the combined treatment showed markedly increased activities of SOD, CAT, and MDA compared to those treated with Cd alone. The effect addition indices (EAIs), ranging from 1.00 to 2.23, unequivocally demonstrated a synergistic effect of the combined treatments. Interestingly, relocating the earthworms to clean soil did not mitigate the observed adverse effects. These findings underscore the increased risk posed by the Cd-SMX complex to terrestrial invertebrates in agricultural areas.

Deciphering the mechanistic role of Bacillus paramycoides (PM51) and Bacillus tequilensis (PM52) in bio-sorption and phyto-assimilation of Cadmium via Linum usitatissimum L. Seedlings.

Three Cd2+ resistant bacterium's minimal inhibition concentrations were assessed and their percentages of Cd2+ accumulation were determined by measurements using an atomic absorption spectrophotometer (AAS). The results revealed that two isolates Bacillus paramycoides (PM51) and Bacillus tequilensis (PM52), identified by 16S rDNA gene sequencing, showed a higher percentage of Cd2+ accumulation i.e., 83.78% and 81.79%, respectively. Moreover, both novel strains can tolerate Cd2+ levels up to 2000 mg/L isolated from district Chakwal. Amplification of the czcD, nifH, and acdS genes was also performed. Batch bio-sorption studies revealed that at pH 7.0, 1 g/L of biomass, and an initial 150 mg/L Cd2+ concentration were the ideal bio-sorption conditions for Bacillus paramycoides (PM51) and Bacillus tequilensis (PM52). The experimental data were fit to Langmuir isotherm measurements and Freundlich isotherm model R2 values of 0.999 for each of these strains. Bio sorption processes showed pseudo-second-order kinetics. The intra-diffusion model showed Xi values for Bacillus paramycoides (PM51) and Bacillus tequilensis (PM52) of 2.26 and 2.23, respectively. Different surface ligands, was investigated through Fourier-transformation infrared spectroscopy (FTIR). The scanning electron microscope SEM images revealed that after Cd2+ adsorption, the cells of both strains became thick, adherent, and deformed. Additionally, both enhanced Linum usitatissimum plant seed germination under varied concentrations of Cd2+ (0 mg/L, 250 mg/L,350 mg/L, and 500 mg/L). Current findings suggest that the selected strains can be used as a sustainable part of bioremediation techniques.

Abiogenic silicon: Interaction with potentially toxic elements and its ecological significance in soil and plant systems.

Abiogenic silicon (Si), though deemed a quasi-nutrient, remains largely inaccessible to plants due to its prevalence within mineral ores. Nevertheless, the influence of Si extends across a spectrum of pivotal plant processes. Si emerges as a versatile boon for plants, conferring a plethora of advantages. Notably, it engenders substantial enhancements in biomass, yield, and overall plant developmental attributes. Beyond these effects, Si augments the activities of vital antioxidant enzymes, encompassing glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD), among others. It achieves through the augmentation of reactive oxygen species (ROS) scavenging gene expression, thus curbing the injurious impact of free radicals. In addition to its effects on plants, Si profoundly ameliorates soil health indicators. Si tangibly enhances soil vitality by elevating soil pH and fostering microbial community proliferation. Furthermore, it exerts inhibitory control over ions that could inflict harm upon delicate plant cells. During interactions within the soil matrix, Si readily forms complexes with potentially toxic metals (PTEs), encapsulating them through Si-PTEs interactions, precipitative mechanisms, and integration within colloidal Si and mineral strata. The amalgamation of Si with other soil amendments, such as biochar, nanoparticles, zeolites, and composts, extends its capacity to thwart PTEs. This synergistic approach enhances soil organic matter content and bolsters overall soil quality parameters. The utilization of Si-based fertilizers and nanomaterials holds promise for further increasing food production and fortifying global food security. Besides, gaps in our scientific discourse persist concerning Si speciation and fractionation within soils, as well as its intricate interplay with PTEs. Nonetheless, future investigations must delve into the precise functions of abiogenic Si within the physiological and biochemical realms of both soil and plants, especially at the critical juncture of the soil-plant interface. This review seeks to comprehensively address the multifaceted roles of Si in plant and soil systems during interactions with PTEs.

Pleiotropic melatonin-mediated responses on growth and cadmium phytoextraction of Brassica napus: A bioecological trial for enhancing phytoremediation of soil cadmium.

Melatonin (MT) has recently gained significant scientific interest, though its mechanism of action in enhancing plant vigor, cadmium (Cd) tolerance, and Cd phytoremediation processes are poorly understood. Therefore, here we investigated the beneficial role of MT in improving growth and Cd remediation potential of rapeseed (Brassica napus). Plants, with or without MT (200 µM L-1), were subjected to Cd stress (30 mg kg1). Without MT, higher Cd accumulation (up to 99%) negatively affected plant growth and developmental feature as well as altered expression of several key genes (DEGs) involved in different molecular pathways of B. napus. As compared to only Cd-stressed counterparts, MT-treated plants exhibited better physiological performance as indicated by improved leaf photosynthetic and gaseous exchange processes (3-48%) followed by plant growth (up to 50%), fresh plant biomass (up to 45%), dry plant biomass (up to 32%), and growth tolerance indices (up to 50%) under Cd exposure. MT application enhanced Cd tolerance and phytoremediation capacity of B. napus by augmenting (1) Cd accumulation in plant tissues and its translocation to above-ground parts (by up to 45.0%), (2) Cd distribution in the leaf cell wall (by up to 42%), and (3) Cd detoxification by elevating phytochelatins (by up to 8%) and metallothioneins (by upto 14%) biosynthesis, in comparison to Cd-treated plants. MT played a protective role in stabilizing hydrogen peroxide and malondialdehyde levels in the tissue of the Cd-treated plants by enhancing the content of osmolytes (proline and total soluble protein) and activities of antioxidant enzymes (SOD, CAT, APX and GR). Transcriptomic analysis revealed that MT regulated 1809 differentially expressed genes (828 up and 981 down) together with 297 commonly expressed DEGs (CK vs Cd and Cd vs CdMT groups) involved in plant-pathogen interaction pathway, protein processing in the endoplasmic reticulum pathway, mitogen-activated protein kinase signaling pathway, and plant hormone signal transduction pathway which ultimately promoted plant growth and Cd remediation potential in the Cd-stressed plants. These results provide insights into the unexplored pleiotropic beneficial action of MT in enhancing in the growth and Cd phytoextraction potential of B. napus, paving the way for developing Cd-tolerant oilseed crops with higher remediation capacity as a bioecological trial for enhancing phytoremediation of hazardous toxic metals in the environment.

Targeting Cd coping mechanisms for stress tolerance in Brassica napus under spiked-substrate system: from physiology to remediation perspective.

Cadmium (Cd) is a prevalent, non-essential, carcinogenic, and hazardous heavy metal that reduces plant productivity and capacity of arable land area around the globe. In the present substrate-based pot study, seedlings of Brassica napus 180015 were grown equidistantly in the spiked-substrate medium for 60 days under increasing concentrations of Cd (0, 10, 20, 30, 40, 50 mg kg-1). Following harvest, the morpho-physio-biochemical, antioxidative, and Cd-induced tolerance responses were evaluated in B. napus under an increasing Cd stress regime. Additionally, these parameters were also investigated to select the plant's threshold tolerance limit for Cd under the spiked-substrate system. B. napus showed dynamic behavior regarding morpho-physio-biochemical attributes, including agronomic features, biomass, photosynthetic pigments, relative water content under increased Cd toxicity. Cd stress-induced hydrogen peroxide (H2O2) production with high MDA contents and passive EL, followed by the orchestration of both enzymatic (SOD, POD, APX, CAT, and GR) and non-enzymatic antioxidants (flavonoids, TPC, TPA, proline, and total soluble protein) up to a certain limit. In addition, Cd-induced stress upregulated transcriptional levels of antioxidative enzyme SOD, POD, APX, GR, and MT encoded genes in B. napus. The increasing trend of Cd accumulation in different tissues at the highest Cd concentration was as follows: root > leaf > stem. In spiked substrate system, B. napus demonstrated improved metal extractability performance and a high potential for phyto-management of low to moderate Cd contamination, implying that this study could be used for integrative breeding programs and decontaminating heavy metals in real contaminated scenarios.Novelty statementThis study provides an insight into Cd-coping mechanisms of oilseed rape involved in alleviating toxicity and simultaneous phyto-management of increasing Cd concentration under spiked substrate system. The current study is the first scientific evidence of using a Cd-spiked soilless substrate medium. The present study will further strengthen our understanding of Cd-instigated positive responses in B. napus. Furthermore, it will provide a useful basis for integrative breeding programs and decontaminating heavy metals in real contaminated scenarios.

A Facile Aptasensor for Instantaneous Determination of Cadmium Ions Based on Fluorescence Amplification Effect of MOPS on FAM-Labeled Aptamer.

Analytical performance and efficiency are two pivotal issues for developing an on-site and real-time aptasensor for cadmium (Cd2+) determination. However, suffering from redundant preparations, fabrications, and incubation, most of them fail to well satisfy the requirements. In this work, we found that fluorescence intensity of 6-carboxyfluorescein(FAM)-labeled aptamer (FAM-aptamer) could be remarkably amplified by 3-(N-morpholino)propane sulfonic acid (MOPS), then fell proportionally as Cd2+ concentration introduced. Importantly, the fluorescence variation occurred immediately after addition of Cd2+, and would keep stable for at least 60 min. Based on the discovery, a facile and ultra-efficient aptasensor for Cd2+ determination was successfully developed. The sensing mechanism was confirmed by fluorescence pattern, circular dichroism (CD) and intermolecular interaction related to pKa. Under the optimal conditions, Cd2+ could be determined rapidly from 5 to 4000 ng mL-1. The detection limit (1.92 ng mL-1) was also lower than the concentration limit for drinking water set by WHO and EPA (3 and 5 ng mL-1, respectively). More than a widely used buffer, MOPS was firstly revealed to have fluorescence amplification effect on FAM-aptamer upon a given context. Despite being sensitive to pH, this simple, high-performance and ultra-efficient aptasensor would be practical for on-site and real-time monitoring of Cd2+.

Small structures with big impact: Multi-walled carbon nanotubes enhanced remediation efficiency in hyperaccumulator Solanum nigrum L. under cadmium and arsenic stress.

With the fast development of nanotechnology, nanomaterials are being increasingly applied for the remediation of contaminated soils. However, few researches have been reported on the complex interactions of carbon nanotubes with heavy metal (loid)s in phytoremediation. Here, we conduct a pot experiment to investigate the effects of multi-walled carbon nanotubes (MWCNTs) on the plant growth and behavior of heavy metal (loid)s in hyperaccumulator-soil system. Cd hyperaccumulator Solanum nigrum L. (S. nigrum) were cultivated in Cadmium (Cd) and Arsenic (As) contaminated soils amended with MWCNTs at 100, 500, and 1000 mg kg-1 for 60 days, respectively. The application of MWCNTs increased the shoot length and plant dry biomass by 5.56%∼25.13% and 5.23%∼27.97%. Whereas, root and leaf growth were inhibited in 1000 mg kg-1 MWCNTs treatments. Meanwhile, MWCNTs at 500 mg kg-1 significantly enhanced the accumulation of heavy metal (loid)s in S. nigrum（18.29% for Cd and 32.47% for As）and alleviated co-contamination induced toxicity, by motivating plant growth, stimulating antioxidant enzymatic activities, and increasing micronutrient content (p < 0.05). The bio-concentration factor of As was decreased (15.31-28.08%) under MWCNTs application, which plays an important role in the alleviation of phytotoxicity. Besides, bioavailable Cd and As were reduced in rhizosphere soils, and the most significant reduction (16.29% for Cd and 8.19% for As) were shown in 500 mg kg-1 MWCNTs treatment. These findings demonstrate that suitable concentration of MWCNTs can enhance remediation efficiency. Our study gives a strong evidence to promote the phytoremediation for co-contaminated soils by using nanomaterials.

Label-Free and Sensitive Determination of Cadmium Ions Using a Ti-Modified Co3O4-Based Electrochemical Aptasensor.

The current work demonstrates an electrochemical aptasensor for sensitive determination of Cd2+ based on the Ti-modified Co3O4 nanoparticles. In this unlabeled system, Ti-modified Co3O4 nanoparticles act as current signal amplifiers modified on the screen-printed carbon electrode (SPCE) surface, while the derivative aptamer of Cd2+ works as a target recognizer. In addition, the sensing is based on the increase in electrochemical probe thionine current signal due to the binding of aptamer to Cd2+ via specific recognition. In the current study, key parameters, including aptamer concentration, pH, and incubation time were optimized, respectively, to ensure sensing performance. Cyclic voltammetry was used not only to characterize each preparation and optimization step, but also to profile the bindings of aptamer to Cd2+. Under optimal conditions, Cd2+ can be determined in a linear range of 0.20 to 15 ng/mL, with a detection limit of 0.49 ng/mL, significantly below the maximum concentration limit set by the U.S. Environmental Protection Agency. Based on comparative analysis and the results of recovery test with real samples, this simple, label-free but highly selective method has considerable potential and thus can be used as an in-situ environmental monitoring platform for Cd2+ testing.

Two plant growth promoting bacterial Bacillus strains possess different mechanisms in adsorption and resistance to cadmium.

Microorganisms are promising biosorbents for decontaminating cadmium-polluted soil or water systems, but the underlying remediation mechanisms are still unclear. In this study, the cadmium biosorption mechanisms and capabilities of plant growth-promoting microorganisms (Bacillus megaterium NCT-2 and Bacillus paranthracis NT1) were investigated. Batch biosorption experiments showed that the optimal biosorption conditions for B. megaterium NCT-2 and B. paranthracis NT1 were pH 6.0, a biomass dosage of 1.0 g L-1, and an initial Cd2+ concentration of 10 mg L-1, and pH 8.0, a biomass dosage of 1.0 g L-1, and an initial Cd2+ concentration of 10 mg L-1, respectively. The biosorption processes of both biosorbents were well described by the pseudo-second order kinetic model, which indicated that the biosorption of Cd2+ was mainly chemisorption. The intracellular accumulation portion of adsorbed Cd2+ in B. megaterium NCT-2 was much higher than in B. paranthracis NT1 (43.11% and 3.25%, respectively), which resulted in the lower cadmium tolerance (14 mg L-1 and 280 mg L-1, respectively) and higher cadmium removal efficiency (46.79% and 20.45%, respectively) of B. megaterium NCT-2 compared to B. paranthracis NT1. SEM-EDS and FTIR analysis suggested the probable interactions of Cd2+ with the biosorbent surface ligands, such as -OH, -NH, -SO3, CO and -COOH during surface adsorption. Results of qRT-PCR illustrated that the difference in cadmium resistant mechanism and adsorption performance between B. megaterium NCT-2 and B. paranthracis NT1 may be regulated by the genes cadA, zitB, khtT, and bshA and cadA, trkA, czcD, and bshA, respectively. Our results revealed that these two biosorbents have the potential for further use in the development of cadmium remediation technologies and could provide insight into the mechanisms of cadmium biosorption.

Characterization of a novel cotton MYB gene, GhMYB108-like responsive to abiotic stresses.

Transcriptional factors are the major regulators of plant signaling pathways in response to environmental stresses i.e., drought, salinity and cold. Hereby, the GhMYB108-like was characterized to determine whether it regulate these stresses. The GhMYB108-like cDNA consisted of 1107 base pairs (bp) with 807 open reading frame encoded a protein of 268 amino acids. Its isoelectric point and molecular weight are 5.51 and 30.3 kDa respectively. Phylogenetic analysis and online databases revealed that GhMYB108-like proteins are closely related with the Arabidopsis thaliana MYB2. Important cis-elements were detected in the promotor region of GhMYB108-like responding to stresses and phytohormones. The 3D structure of GhMYB108-like protein has been predicted. In addition, various physico-chemical properties of GhMYB108-like have been determined. Subcellular localization confirmed that GhMYB108-like are nuclear localized protein. Quantitative expression analysis showed that polyethylene glycol and salt treatments significantly induced the expression of GhMYB108-like. Overall, our findings suggest that GhMYB108-like is an important gene that would plays important regulatory role in response to drought and salt stresses.

Optimization of NPK fertilization combined with phytoremediation of cadmium contaminated soil by orthogonal experiment.

In the current experiment, influence of NPK composition on the Cd contaminated soil-plant (Solanum nigrum L.) system as well as the phytoremediation efficiency were comprehensively studied. The composition of NPK was optimized for a sustainable phytoremediation and simultaneous agronomic technique in Cd-contaminated soil by orthogonal (L14) experiment, aimed to achieve plant productivity and maximum phytoremediation potential enhancement. Results showed that different treatments of NPK composition enhanced soil properties including saccharase, urease, catalase and acid phosphatase activities as compared to the control treatment, however, soil pH was slightly decreased by 3.64%~6.67% with different composition of NPK treatments. Plant biomass and Cd concentration in the aboveground part (stem and leaves) of S. nigrum were significantly (P < 0.05) enhanced by 14.19%~48.97% and 38.50%~127.15% as compared to control plants with the addition of NPK fertilizers having different composition. Meanwhile, with the application of NPK fertilizer root/shoot Cd ratio and translocation factor (TF) was significantly decreased, however, bioconcentration factor (BCF) was increased as compared to control. Additionally, different composition of NPK fertilizers significantly increased photosynthetic pigments (chlorophyll a, chlorophyll b, total chlorophyll, and carotenoids) and soluble protein in comparison to control. The activities of antioxidant enzymes in S. nigrum including ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and glutathione reductase (GR) were increased, while malonaldehyde (MDA) and proline contents were decreased. Principal component analysis (PCA) showed that N3P2K2 treatment had the highest comprehensive score amongst other studied treatments of NPK composition, owing to its optimal composition for the investigated soil-S. nigrum system. Moreover, it was found that optimal composition (N3P2K2) of fertilizer resulted in increase of the plant resistance to Cd and the efficiency of phytoextraction. Therefore, it is suggested to all the small-holder famers and scientific community that precise composition of NPK fertilizer should be utilized according to soil properties, environmental conditions and plant requirements under Cd-stress condition in order to achieve maximum biomass, Cd uptake efficiency as well phytoremediation potential in moderately Cd contaminated soil.