Heavy Metal Detoxification Research Articles

Hazardous or Advantageous: Uncovering the Roles of Heavy Metals and Humic Substances in Shilajit (Phyto-mineral) with Emphasis on Heavy Metals Toxicity and Their Detoxification Mechanisms.

Shilajit is a phyto-mineral diffusion and semi-solid matter used as traditional medicine with extraordinary health benefits. This study provides a comprehensive data on Shilajit with emphasis on heavy metal profile, associated toxicities, and metal detoxification mechanisms by humic substances present in Shilajit. Data was searched across papers and traditional books using Google Scholar, PubMed, Science Direct, Medline, SciELO, Web of Science, and Scopus as key scientific databases. Findings showed that Shilajit is distributed in almost 20 regions of the world with uses against 20 health problems as traditional medicine. With various humic substances, almost 11 biological activities were reported in Shilajit. This phyto-mineral diffusion possesses around 65 heavy metals including the toxic heavy metals like Cu, Al, Pb, As, Cd, and Hg. However, humic substances in Shilajit actively detoxify around 12 heavy metals. The recommended levels of heavy metals by WHO and FDA in herbal drugs is 0.20 and 0.30ppm for Cd, 1ppm for Hg, 10.00ppm for As and Pb, 20ppm for Cu, and 50ppm for Zn. The levels of reported metals in Shilajit were found to be lower than the permissible limits set by WHO and FDA, except in few studies where exceeded levels were reported. Shilajit consumption without knowing permissible levels of metals is not safe and could pose serious health problems. Although the humic substances and few metals in Shilajit are beneficial in terms of chelating toxic heavy metals, the data on metal detoxification still needs to be clarified.

Novel endolithic bacteria of phylum Chloroflexota reveal a myriad of potential survival strategies in the Antarctic desert.

The ice-free McMurdo Dry Valleys of Antarctica are dominated by nutrient-poor mineral soil and rocky outcrops. The principal habitat for microorganisms is within rocks (endolithic). In this environment, microorganisms are provided with protection against sub-zero temperatures, rapid thermal fluctuations, extreme dryness, and ultraviolet and solar radiation. Endolithic communities include lichen, algae, fungi, and a diverse array of bacteria. Chloroflexota is among the most abundant bacterial phyla present in these communities. Among the Chloroflexota are four novel classes of bacteria, here named Candidatus Spiritibacteria class. nov. (=UBA5177), Candidatus Martimicrobia class. nov. (=UBA4733), Candidatus Tarhunnaeia class. nov. (=UBA6077), and Candidatus Uliximicrobia class. nov. (=UBA2235). We retrieved 17 high-quality metagenome-assembled genomes (MAGs) that represent these four classes. Based on genome predictions, all these bacteria are inferred to be aerobic heterotrophs that encode enzymes for the catabolism of diverse sugars. These and other organic substrates are likely derived from lichen, algae, and fungi, as metabolites (including photosynthate), cell wall components, and extracellular matrix components. The majority of MAGs encode the capacity for trace gas oxidation using high-affinity uptake hydrogenases, which could provide energy and metabolic water required for survival and persistence. Furthermore, some MAGs encode the capacity to couple the energy generated from H2 and CO oxidation to support carbon fixation (atmospheric chemosynthesis). All encode mechanisms for the detoxification and efflux of heavy metals. Certain MAGs encode features that indicate possible interactions with other organisms, such as Tc-type toxin complexes, hemolysins, and macroglobulins.IMPORTANCEThe ice-free McMurdo Dry Valleys of Antarctica are the coldest and most hyperarid desert on Earth. It is, therefore, the closest analog to the surface of the planet Mars. Bacteria and other microorganisms survive by inhabiting airspaces within rocks (endolithic). We identify four novel classes of phylum Chloroflexota, and, based on interrogation of 17 metagenome-assembled genomes, we predict specific metabolic and physiological adaptations that facilitate the survival of these bacteria in this harsh environment-including oxidation of trace gases and the utilization of nutrients (including sugars) derived from lichen, algae, and fungi. We propose that such adaptations allow these endolithic bacteria to eke out an existence in this cold and extremely dry habitat.

Biosorption of halophilic fungal melanized membrane – PUR/melanin polymer for heavy metal detoxification with electrospinning technology

ABSTRACT Eradication of heavy metal pollution has become the prime priority over the conservation of water resources in the upcoming era. Herein, the study involved the halophilic fungal melanin from Curvularia lunata showed a promising biosorbent for the removal of toxic heavy metals which shows eco-friendly, cost-effective, high stability, and adsorbent efficiency. Polyurethane blended with fungal melanin polymers, makes polymeric nanofibrous membranes through electrospinning techniques. BET isotherms revealed the raw fungal melanin holds a surface area of 3.54 m2/g exhibiting type IV isotherms. BJH results in a total pore volume of 5.79 cc/g with a pore diameter of 6.54 ± 1 nm for pores smaller than 4544.8 Å. Exhibits Eumelanin properties were characterized by FE – SEM and FTIR functional elements. ICPMS confirmed the metal adsorption proficiency on both raw and melanized membranes before and after treatments. Over 17 heavy metals, Ni2+ were adsorbed with 100% efficiency by raw melanin alone with 42.48 µg/L of Ni2+ concentration in the water sample, whereas, Cu2+, Zn2+, Co2+, Cr2+, Pb2+, Mn2+, Al3+, Mo6+, Sb3+, Ba2+, Fe2+, and Mg2+ stands next with 99%. In this study, gentle/simple application of raw fungal melanin (without PUR tailored) can detoxify the maximum concentration of heavy metals present in the water bodies which are further used for irrigation and even drinking purposes. This mycoremediation approach can be easily adapted to industrial production than other high-performance membrane materials with minimal process modification, making it a promising strategy for improving the adsorption properties used in various applications after still furthermore investigation.

Unlocking nature’s clean-up crew: marine Bacillus species as agents of heavy metal detoxification

Unlocking nature’s clean-up crew: marine Bacillus species as agents of heavy metal detoxification

Enhanced use of chemical fertilizers and mitigation of heavy metal toxicity using biochar and the soil fungus Bipolaris maydis AF7 in rice: Genomic and metabolomic perspectives

Chemical fertilizers are the primary source of crop nutrition; however, their increasing rate of application has created environmental hazards, such as heavy metal toxicity and eutrophication. The synchronized use of chemical fertilizers and eco-friendly biological tools, such as microorganisms and biochar, may provide an efficient foundation to promote sustainable agriculture. Therefore, the current study aimed to optimize the nutrient uptake using an inorganic fertilizer, sulfate of potash (SOP) from the plant growth-promoting fungus Bipolaris maydis AF7, and biochar under heavy metal toxicity conditions in rice. Bioassay analysis showed that AF7 has high resistance to heavy metals and a tendency to produce gibberellin, colonize the fertilizer, and increase the intake of free amino acids. In the plant experiment, the co-application of AF7 +Biochar+MNF+SOP significantly lowered the heavy metal toxicity, enhanced the nutrient uptake in the rice shoots, and improved the morphological attributes (total biomass). Moreover, the co-application augmented the glucose and sucrose levels, whereas it significantly lowered the endogenous phytohormone levels (salicylic acid and jasmonic acid) in the rice shoots. The increase in nutrient content aligns with the higher expression of the OsLSi6, PHT1, and OsHKT1 genes. The plant growth traits and heavy metal tolerance of AF7 were validated by whole-genome sequencing that showed the presence of the heavy metal tolerance and detoxification protein, siderophore iron transporter, Gibberellin cluster GA4 desaturase, and DES_1 genes, as well as others that regulate glucose, antioxidants, and amino acids. Because the AF7 +biochar+inorganic fertilizer works synergistically, nutrient availability to the crops could be improved, and heavy metal toxicity and environmental hazards could be minimized.

Types of Bioremediation and their Applications, in Heavy Metal Detoxification

Heavy metal contamination is the result of various human activities. Increased concentrations of heavy metals are responsible for causing lethal and chronic diseases in humans, affecting the metabolism of plants and causing metal toxicity in the environment. Available conventional techniques for metal detoxification are not found effective and economically feasible. Bioremediation is an ecologically and economically favorable technique that consists of natural biological processes to eliminate toxic heavy metal contaminants. Various microorganisms, fungi and plants can be used for heavy metal detoxification. This paper gives an idea about the concept, types, and principles of bioremediation.

Characterization of a Nicotiana tabacum phytochelatin synthase 1 and its response to cadmium stress.

Phytochelatin synthase (PCS) is a critical enzyme involved in heavy metal detoxification in organisms. In this study, we aim to comprehensively investigate the molecular and functional characteristics of the PCS1 gene from Nicotiana tabacum by examining its enzymatic activity, tissue-specific expression pattern, Cd-induced expression, as well as the impact on Cd tolerance and accumulation. The results demonstrated that the amino acid sequence of NtPCS1 shared a highsimilarity in its N-terminal region with PCS from other species. The enzymaticactivity of NtPCS1 was found to be enhanced in the order Ag2+ > Cd2+ > Cu2+ > Pb2+ > Hg2+ > Fe2+ > Zn2+. In addition, RT-PCR data indicated that NtPCS1 gene is constitutively expressed, with the highest expression observed in flowers, and that its transcript levels are up-regulated by CdCl2. When tobacco overexpressing NtPCS1 (PCS1 lines) were grown under CdCl2 stress, they produced more phytochelatins (PCs) than WT plants, but this did not result in increased Cd accumulation. However, in a root growth assay, the PCS1 lines exhibited hypersensitivity to Cd. The overexpression of NtPCS1 itself does not appear to be the primary cause of this heightened sensitivity to Cd, as the Arabidopsis thaliana Atpcs1 mutant overexpressing NtPCS1 actually exhibited enhanced tolerance to Cd. Furthermore, the addition of exogenous glutathione (GSH) progressively reduced the Cd hypersensitivity of the PCS1 lines, with the hypersensitivity even being completely eliminated. Surprisingly, the application of exogenous GSH led to a remarkably enhanced Cd accumulation in the PCS1 lines. This study enriches our understanding of the molecular function of the NtPCS1 gene and suggests a promising avenue for Cd tolerance through the heterologous expression of PCS genes in different species.

Bacterial tolerance and detoxification of cyanide, arsenic and heavy metals: Holistic approaches applied to bioremediation of industrial complex wastes.

Cyanide is a highly toxic compound that is found in wastewaters generated from different industrial activities, such as mining or jewellery. These residues usually contain high concentrations of other toxic pollutants like arsenic and heavy metals that may form different complexes with cyanide. To develop bioremediation strategies, it is necessary to know the metabolic processes involved in the tolerance and detoxification of these pollutants, but most of the current studies are focused on the characterization of the microbial responses to each one of these environmental hazards individually, and the effect of co-contaminated wastes on microbial metabolism has been hardly addressed. This work summarizes the main strategies developed by bacteria to alleviate the effects of cyanide, arsenic and heavy metals, analysing interactions among these toxic chemicals. Additionally, it is discussed the role of systems biology and synthetic biology as tools for the development of bioremediation strategies of complex industrial wastes and co-contaminated sites, emphasizing the importance and progress derived from meta-omic studies.

BcaSOD1 enhances cadmium tolerance in transgenic Arabidopsis by regulating the expression of genes related to heavy metal detoxification and arginine synthesis

Cadmium (Cd), which is a nonessential heavy metal element for organisms, can have a severe impact on the growth and development of organisms that absorb excessive Cd. Studies have shown that Brassica carinata, a semiwild oil crop, has strong tolerance to various abiotic stresses, and RNA-seq has revealed that the B. carinata superoxide dismutase gene (BcaSOD1) likely responds to Cd stress. To elucidate the BcaSOD1 function involved in tolerance of Cd stress, we cloned the coding sequences of BcaSOD1 from a purple B. carinata accession and successfully transferred it into Arabidopsis thaliana. The subcellular localization results demonstrated that BcaSOD1 was primarily located in the plasma membrane, mitochondria and nucleus. Overexpression of BcaSOD1 in transgenic Arabidopsis (OE) effectively decreased the toxicity caused by Cd stress. Compared to the WT (wild type lines), the OE lines exhibited significantly increased activities of antioxidant enzymes (APX, CAT, POD, and SOD) after exposure to 2.5 mM CdCl2. The Cd content of underground (root) in the OE line was dominantly higher than that in the WT; however, the Cd content of aboveground (shoot) was comparable between the OE and WT types. Moreover, the qRT‒PCR results showed that several heavy metal detoxification-related genes (AtIREG2, AtMTP3, AtHMA3, and AtNAS4) were significantly upregulated in the roots of OE lines under Cd treatment, suggesting that these genes are likely involved in Cd absorption in the roots of OE lines. In addition, both comparable transcriptome and qRT-PCR analyses revealed that exogenous BcaSOD1 noticeably facilitates detoxification by stimulating the expression of two arginine (Arg) biosynthesis genes (AtGDH1 and AtGDH2) while inhibiting the expression of AtARGAH1, a negative regulator in biosynthesis of Arg. The Arg content was subsequently confirmed to be significantly enhanced in OE lines under Cd treatment, indicating that BcaSOD1 likely strengthened Cd tolerance by regulating the expression of Arg-related genes. This study demonstrates that BcaSOD1 can enhance Cd tolerance and reveals the molecular mechanism of this gene, providing valuable insights into the molecular mechanism of Cd tolerance in plants.

Culturable Bacterial Endophytes of Wild White Poplar (Populus alba L.) Roots: A First Insight into Their Plant Growth-Stimulating and Bioaugmentation Potential.

The white poplar (Populus alba L.) has good potential for a green economy and phytoremediation. Bioaugmentation using endophytic bacteria can be considered as a safe strategy to increase poplar productivity and its resistance to toxic urban conditions. The aim of our work was to find the most promising strains of bacterial endophytes to enhance the growth of white poplar in unfavorable environmental conditions. To this end, for the first time, we performed whole-genome sequencing of 14 bacterial strains isolated from the tissues of the roots of white poplar in different geographical locations. We then performed a bioinformatics search to identify genes that may be useful for poplar growth and resistance to environmental pollutants and pathogens. Almost all endophytic bacteria obtained from white poplar roots are new strains of known species belonging to the genera Bacillus, Corynebacterium, Kocuria, Micrococcus, Peribacillus, Pseudomonas, and Staphylococcus. The genomes of the strains contain genes involved in the enhanced metabolism of nitrogen, phosphorus, and metals, the synthesis of valuable secondary metabolites, and the detoxification of heavy metals and organic pollutants. All the strains are able to grow on media without nitrogen sources, which indicates their ability to fix atmospheric nitrogen. It is concluded that the strains belonging to the genus Pseudomonas and bacteria of the species Kocuria rosea have the best poplar growth-stimulating and bioaugmentation potential, and the roots of white poplar are a valuable source for isolation of endophytic bacteria for possible application in ecobiotechnology.

Dynamics of Humic Acid, Silicon, and Biochar under Heavy Metal, Drought, and Salinity with Special Reference to Phytohormones, Antioxidants, and Melatonin Synthesis in Rice.

This study aimed to develop a biostimulant formulation using humic acid (HA), silicon, and biochar alone or in combination to alleviate the lethality induced by combined heavy metals (HM-C; As, Cd, and Pb), drought stress (DS; 30-40% soil moisture), and salt stress (SS; 150 mM NaCl) in rice. The results showed that HA, Si, and biochar application alone or in combination improved plant growth under normal, DS, and SS conditions significantly. However, HA increased the lethality of rice by increasing the As, Cd, and Pb uptake significantly, thereby elevating lipid peroxidation. Co-application reduced abscisic acid, elevated salicylic acid, and optimized the Ca2+ and Si uptake. This subsequently elevated the K+/Na+ influx and efflux by regulating the metal ion regulators (Si: Lsi1 and Lsi2; K+/Na+: OsNHX1) and increased the expressions of the stress-response genes OsMTP1 and OsNramp in the rice shoots. Melatonin synthesis was significantly elevated by HM-C (130%), which was reduced by 50% with the HA + Si + biochar treatment. However, in the SS- and DS-induced crops, the melatonin content showed only minor differences. These findings suggest that the biostimulant formulation could be used to mitigate SS and DS, and precautions should be taken when using HA for heavy metal detoxification.

Plasmid-encoded lactose metabolism and mobilized colistin resistance (mcr-9) genes in Salmonella enterica serovars isolated from dairy facilities in the 1980s.

Horizontal gene transfer by plasmids can confer metabolic capabilities that expand a host cell's niche. Yet, it is less understood whether the coalescence of specialized catabolic functions, antibiotic resistances and metal resistances on plasmids provides synergistic benefits. In this study, we report whole-genome assembly and phenotypic analysis of five Salmonella enterica strains isolated in the 1980s from milk powder in Munich, Germany. All strains exhibited the unusual phenotype of lactose-fermentation and encoded either of two variants of the lac operon. Surprisingly, all strains encoded the mobilized colistin resistance gene 9 (mcr-9), long before the first report of this gene in the literature. In two cases, the mcr-9 gene and the lac locus were linked within a large gene island that formed an IncHI2A-type plasmid in one strain but was chromosomally integrated in the other strain. In two other strains, the mcr-9 gene was found on a large IncHI1B/IncP-type plasmid, whereas the lac locus was encoded on a separate chromosomally integrated plasmidic island. The mcr-9 sequences were identical and genomic contexts could not explain the wide range of colistin resistances exhibited by the Salmonella strains. Nucleotide variants did explain phenotypic differences in motility and exopolysaccharide production. The observed linkage of mcr-9 to lactose metabolism, an array of heavy-metal detoxification systems, and other antibiotic resistance genes may reflect a coalescence of specialized phenotypes that improve the spread of colistin resistance in dairy facilities, much earlier than previously suspected.

Dietary Wood and Activated Charcoal Improved Ammonium Removal, Heavy Metals Detoxification, Growth Performance, Blood Biochemistry, Carcass Traits, and Histopathology of European Seabass.

A 120-day growth trial was completed to assess rearing water quality and fish performance in terms of growth, feed efficacy, digestive enzymes, immunity, and antioxidant activity of seabass fed an experimental diet (ED) supplemented with commercial wood charcoal (WC) and activated wood charcoal (AC). Three levels (0, 10, and 20 g) of WC and AC were administered, representing five treatments: control (CD) fish-fed ED without additives, (WC-1) fish-fed ED containing 10 g kg-1 WC, (WC-2) fish-fed ED containing 20 g kg-1 WC, (AC-1) fish-fed ED containing 10 g kg-1 AC, and (AC-2) fish-fed ED containing 20 g kg-1 AC. Three hundred fish (60.12 ± 0.20 g/fish) were stocked in 15 cement tanks (4.0 m × 2.0 m × 1.2 m, water volume 5 m3 each) at 20 fish/tank and a daily feed ration of 3% of body weight. Results revealed significant improvements with increased growth variables (final weight, weight gain, and specific growth rate), decreased FCR, and decreased ammonia levels and heavy metals (Cu, Cd, Fe, Mn, and Zn) content in rearing water, muscle, and liver with fish fed WC and AC supplemented diets. Furthermore, considerable improvements in digestive enzymes, immunity, and antioxidant activity, with enhanced kidneys, liver, intestines, gills, and spleen. Fish fed the WC-1 diet had a higher final weight (171.90 g), better FCR (1.25), and improved internal organs than the other groups.

Untying the regulatory roles of miRNAs in CuO-NPs stress response mechanism in maize: A genome-wide sRNA transcriptome analysis

MicroRNAs (miRNAs), a group of tiny non-coding RNAs play pivotal role in plant responses to environmental stress. The present small RNA transcriptome study aims to untie the role of miRNAs in CuO-NPs stress adaptation in maize seedlings. Restricted seedling growth, enhanced ROS generation and higher membrane damage were recorded under CuO-NPs [<50 nm, 8 mM] treatment. Deep sequencing reveals 7 up- and 36 down-regulated known miRNAs from CuO-NPs challenged leaves. Gene ontology study demonstrates involvement of CuO-NPs responsive miRNAs in a variety of biological processes including plant growth (miR159a, miR159b), redox homeostasis (miR156e, miR395a), detoxification of heavy metals (miR156e, miR827), signal transduction (miR156e, miR156d), and cell signalling (miR167b-3p, miR393a). Enhanced transcriptional abundance of ABC transporter G family member 41 isoform X2 and HM-associated isoprenylated plant protein 45 isoform X1 might be involved in sequestration and detoxification of excess Cu, essential for metal homeostasis in maize. The miR528-5p mediated up-regulation of superoxide dismutase does not give much protection against CuO-NPs induced oxidative stress damages as evident after histochemical staining with NBT. Moreover, CuO-NPs stress mediated down regulation of miR396 could be an underlying cause of the restricted seedling growth. Taken together, our findings provide insights into the miRNA-guided stress regulatory networks involved in plant's adaptive responses to CuO-NPs stress.

Heavy metals mitigation and growth promoting effect of endophytic Agrococcus terreus (MW 979614) in maize plants under zinc and nickel contaminated soil.

Heavy metals such as iron, copper, manganese, cobalt, silver, zinc, nickel, and arsenic have accumulated in soils for a long time due to the dumping of industrial waste and sewage. Various techniques have been adapted to overcome metal toxicity in agricultural land but utilizing a biological application using potential microorganisms in heavy metals contaminated soil may be a successful approach to decontaminate heavy metals soil. Therefore, the current study aimed to isolate endophytic bacteria from a medicinal plant (Viburnum grandiflorum) and to investigate the growth-promoting and heavy metal detoxification potential of the isolated endophytic bacteria Agrococus tereus (GenBank accession number MW 979614) under nickel and zinc contamination. Zinc sulfate and nickel sulfate solutions were prepared at the rate of 100 mg/kg and 50 mg/kg in sterilized distilled water. The experiment was conducted using a completely random design (CRD) with three replicates for each treatment. Inoculation of seeds with A. tereus significantly increased the plant growth, nutrient uptake, and defense system. Treatment T4 (inoculated seeds), T5 (inoculated seeds + Zn100 mg/kg), and T6 (inoculated seeds + Ni 100 mg/kg) were effective, but T5 (inoculated seeds + Zn100 mg/kg) was the most pronounced and increased shoot length, root length, leaf width, plant height, fresh weight, moisture content, and proline by 49%, 38%, 89%, 31%, 113%, and 146%, respectively. Moreover the antioxidant enzymes peroxidase and super oxidase dismutase were accelerated by 211 and 68% in contaminated soil when plants were inoculated by A. tereus respectively. Similarly the inoculation of A. tereus also enhanced maize plants' absorption of Cu, Mn, Ni, Na, Cr, Fe, Ca, Mg, and K significantly. Results of the findings concluded that 100 mg/kg of Zn and Ni were toxic to maize growth, but seed inoculation with A. tereus helped the plants significantly in reducing zinc and nickel stress. The A. tereus strain may be employed as a potential strain for the detoxification of heavy metals.

Bio-mediated detoxification of heavy metal contaminated soil and phytotoxicity reduction using novel strain of Brevundimonas vancanneytii SMA3

Heavy metals pose a serious environmental threat on a global scale due to their toxicity towards livings. Therefore, removing harmful metals from the environment has become more challenging in recent years. The objective of this study is to isolate, examine, and characterize naturally existing bacteria that possess the ability to mitigate and detoxify heavy metals such as cadmium, mercury, and lead. The selected bacteria SMA3 actively demonstrated metal tolerance during screening and was then employed for biosorption study using a lab-scale technique. The bacterium belonged to Brevundimonas sp., according to 16 S rRNA analysis. To enhance the removal efficiency of SMA3, response surface methodology (RSM) was employed, resulting in the identification of optimized conditions (pH 7, temperature 30 °C and shaking speed 120 rpm) for achieving maximum removal percentage (69.5 % of Cd, 58.6 % of Hg, and 85.1 % of Pb) within 72 h. The structural changes induced by microbial treatment were demonstrated by comparing the findings of FESEM images and FTIR spectra confirming the disappearance of C ^ C, C]O peaks along with C]O, C–O–C, C–H, and O–H bond destabilization following bioaccumulation. Moreover, in terms of phytotoxicity evaluation, it was observed that the treated soil, containing both heavy metals and the selected potent bacterial strain, exhibited reduced toxicity, resulting in improved germination and growth parameters for the seeds of Solanum lycopersicum (tomato plant). Overall, the selected bacterial strain demonstrated its potential for effectively removing multiple metals from the metal contaminated environment.

Proteome insights of citric acid-mediated cadmium toxicity tolerance in Brassica napus L.

Cadmium (Cd) is a toxic substance that is uptake by plants from soils, Cd easily transfers into the food chain. Considering global food security, eco-friendly, cost-effective, and metal detoxification strategies are highly demandable for sustainable food crop production. The purpose of this study was to investigate how citric acid (CA) alleviates or tolerates Cd toxicity in Brassica using a proteome approach. In this study, the global proteome level was significantly altered under Cd toxicity with or without CA supplementation in Brassica. A total of 4947 proteins were identified using the gel-free proteome approach. Out of these, 476 proteins showed differential abundance between the treatment groups, wherein 316 were upregulated and 160 were downregulated. The gene ontology analysis reveals that differentially abundant proteins were involved in different biological processes including energy and carbohydrate metabolism, CO2 assimilation and photosynthesis, signal transduction and protein metabolism, antioxidant defense, heavy metal detoxification, plant development, and cytoskeleton and cell wall structure in Brassica leaves. Interestingly, several candidate proteins such as superoxide dismutase (A0A078GZ68) L-ascorbate peroxidase 3 (A0A078HSG4), glutamine synthetase (A0A078HLB2), glutathione S-transferase DHAR1 (A0A078HPN8), glutamine synthetase (A0A078HLB2), cysteine synthase (A0A078GAD3), S-adenosylmethionine synthase 2 (A0A078JDL6), and thiosulfate/3-mercaptopyruvate sulfur transferase 2 (A0A078H905) were involved in antioxidant defense system and sulfur assimilation-involving Cd-detoxification process in Brassica. These findings provide new proteome insights into CA-mediated Cd-toxicity alleviation in Brassica, which might be useful to oilseed crop breeders for enhancing heavy metal tolerance in Brassica using the breeding program, with sustainable and smart Brassica production in a metal-toxic environment.

Investigating the growth characteristics, oxidative stress, and metal absorption of chickpea (Cicer arietinum L.) under cadmium stress and in silico features of HMAs proteins

Heavy metal contamination can have a strong effect on the morphological and physiological characteristics of plants. In the present study, Cicer arietinum L. (chickpea) was exposed to different concentrations of cadmium (control, 2, 4, 8 μg Cd g-1 perlite) and the effect on plant growth and antioxidant enzymes were evaluated. The observed morphological changes in chickpea plant included stunted growth, reduced root system development and plant color change. A significant increase in enzyme activity of peroxidase, superoxide dismutase, catalase, and ascorbate peroxidase was observed at 4 μg Cd g-1 perlite, with a subsequent decrease when concentration was increased to 8 μg Cd g-1 perlite in the leaves of the plants. The highest cadmium levels were determined at a concentration of 8 μg Cd g-1 perlite. With the addition of 2 μg Cd g-1 perlite, manganese uptake in the aboveground part of the plant increased significantly, but then decrease at higher cadmium concentrations. In addition, zinc and copper levels decrease in the presence of cadmium. These results indicate that chickpea has a relatively high adsorption capacity for cadmium in aboveground tissues and special precautions should be taken when growing chickpea. In silico analysis led to the identification of 13 heavy metal ATPases (HMAs) in chickpea. These proteins contain 130 to 1032 amino acids with 3 to 18 exons. They are involved in the transfer of cadmium and zinc and help in heavy metal detoxification of plants. Bioinformatics studies have been conducted to better understand the mechanism by which the plant is able to combat heavy metal stress.

Genetic analysis of the variation for mineral accumulation in the leaves and seeds of natural germplasm of Brassica rapa L. (AA) and the its derived forms extracted from an allotetraploid B.juncea L.(AABB)

Brassica rapa L. (2n = 20; AA) is a vegetable and oilseed crop that is grown all over the world. Its leaves, shoots, and seeds store significant amounts of minerals. We used inductively coupled plasma-optical emission spectroscopy (ICP-OES) to determine the concentrations of eleven minerals in the leaves and seeds of 195 advanced generation inbred lines, of which 92 represented natural (NR) B. rapa and the remaining 103 were derived (DR) from a set of mother genotypes originally extracted from an allotetraploid B. juncea (2n = 36; AABB). The inbred lines differed for the composition of leaf and seed minerals. Leaf concentrations of N, K, Zn, and Se were higher in the DR subpanel as compared to NR subpanel, along with high seed accumulations of K and Se. DArT genotyping and genome wide association mapping led to the identification of SNPs associated with leaf and seed mineral compositions. Chromosomes A03, A05, and A10 harboured the most associated loci. Annotations of the regions adjacent to respective GWAS peaks allowed prediction of genes known for acquisition, transport, and accumulation of minerals and heavy metal detoxification. Transcriptome analysis revealed differential expression patterns of the predicted candidates, with most genes either down-regulated in derived genotypes relative to natural forms or their expression being comparable between the two. General downregulation may be a consequence of extracting B. rapa from allotetraploid B. juncea through genome resection. Some of the identified SNPs may be used as DNA markers for breeding programmes designed to modify the leaf and seed mineral compositions.

Genome Analysis and Potential Ecological Functions of Members of the Genus Ensifer from Subsurface Environments and Description of Ensifer oleiphilus sp. nov.

The current work deals with genomic analysis, possible ecological functions, and biotechnological potential of two bacterial strains, HO-A22T and SHC 2-14, isolated from unique subsurface environments, the Cheremukhovskoe oil field (Tatarstan, Russia) and nitrate- and radionuclide-contaminated groundwater (Tomsk region, Russia), respectively. New isolates were characterized using polyphasic taxonomy approaches and genomic analysis. The genomes of the strains HO-A22T and SHC 2-14 contain the genes involved in nitrate reduction, hydrocarbon degradation, extracellular polysaccharide synthesis, and heavy metal detoxification, confirming the potential for their application in various environmental biotechnologies. Genomic data were confirmed by cultivation studies. Both strains were found to be neutrophilic, chemoorganotrophic, facultatively anaerobic bacteria, growing at 15-33 °C and 0-1.6% NaCl (w/v). The 16S rRNA gene sequences of the strains were similar to those of the type strains of the genus Ensifer (99.0-100.0%). Nevertheless, genomic characteristics of strain HO-A22T were below the thresholds for species delineation: the calculated average nucleotide identity (ANI) values were 83.7-92.4% (<95%), and digital DNA-DNA hybridization (dDDH) values were within the range of 25.4-45.9% (<70%), which supported our conclusion that HO-A22T (=VKM B-3646T = KCTC 92427T) represented a novel species of the genus Ensifer, with the proposed name Ensifer oleiphilus sp. nov. Strain SHC 2-14 was assigned to the species 'Ensifer canadensis', which has not been validly published. This study expanded the knowledge about the phenotypic diversity among members of the genus Ensifer and its potential for the biotechnologies of oil recovery and radionuclide pollution treatment.