Mechanisms Of Cadmium Resistance Research Articles

Genomic and transcriptomic analyses reveal insights into cadmium resistance mechanisms of Cupriavidus nantongensis strain E324

The cadmium-resistant Cupriavidus sp. strain E324 has been previously shown to have a high potential for use in cadmium (Cd) remediation, due to its high capacity for cadmium bioaccumulation. According to the comparative genomic analysis, the strain E324 was most closely related to C. nantongensis X1T, indicating that the strain E324 should be re-identified as C. nantongensis. To unravel the Cd tolerance mechanisms of C. nantongensis strain E324, the transcriptional response of this strain to acute Cd exposure was assessed using RNA-seq-based transcriptome analysis, followed by validation through qRT-PCR. The results showed that the upregulated Differentially Expressed Genes (DEGs) were significantly enriched in categories related to metal binding and transport, phosphate transport, and oxidative stress response. Consistently, we observed significant increases in both the cell wall and intracellular contents of certain essential metals (Cu, Fe, Mn, and Zn) upon Cd exposure. Among these, only the Zn pretreatment resulting in high Zn accumulation in the cell walls could enhance bacterial growth under Cd stress conditions through its role in inhibiting Cd accumulation. Additionally, the promotion of catalase activity and glutathione metabolism upon Cd exposure to cope with Cd-induced oxidative stress was demonstrated. Meanwhile, the upregulation of phosphate transport-related genes upon Cd treatment seems to be the bacterial response to Cd-induced phosphate depletion. Altogether, our findings suggest that these adaptive responses are critical mechanisms contributing to increased Cd tolerance in C. nantongensis strain E324 via the enhancement of metal-chelating and antioxidant capacities of the cells.

Molecular identification of Proteus mirabilis, Vibrio species leading to CRISPR-Cas9 modification of tcpA and UreC genes causing cholera and UTI

Heavy metal accumulation increases rapidly in the environment due to anthropogenic activities and industrialization. The leather and surgical industry produces many contaminants containing heavy metals. Cadmium, a prominent contaminant, is linked to severe health risks, notably kidney and liver damage, especially among individuals exposed to contaminated wastewater. This study aims to leverage the natural cadmium resistance mechanisms in bacteria for bioaccumulation purposes. The industrial wastewater samples, characterized by an alarming cadmium concentration of 29.6 ppm, 52 ppm, and 76.4 ppm—far exceeding the recommended limit of 0.003 ppm—were subjected to screening for cadmium-resistant bacteria using cadmium-supplemented media with CdCl2. 16S rRNA characterization identified Vibrio cholerae and Proteus mirabilis as cadmium-resistant bacteria in the collected samples. Subsequently, the cadmium resistance-associated cadA gene was successfully amplified in Vibrio species and Proteus mirabilis, revealing a product size of 623 bp. Further analysis of the identified bacteria included the examination of virulent genes, specifically the tcpA gene (472 bp) associated with cholera and the UreC gene (317 bp) linked to urinary tract infections. To enhance the bioaccumulation of cadmium, the study proposes the potential suppression of virulent gene expression through in-silico gene-editing tools such as CRISPR-Cas9. A total of 27 gRNAs were generated for UreC, with five selected for expression. Similarly, 42 gRNA sequences were generated for tcpA, with eight chosen for expression analysis. The selected gRNAs were integrated into the lentiCRISPR v2 expression vector. This strategic approach aims to facilitate precise gene editing of disease-causing genes (tcpA and UreC) within the bacterial genome. In conclusion, this study underscores the potential utility of Vibrio species and Proteus mirabilis as effective candidates for the removal of cadmium from industrial wastewater, offering insights for future environmental remediation strategies.

Topless–related 2 conferred cadmium accumulation in wheat

Wheat is a vital food crop that faces threats from various abiotic and biotic stresses. Understanding the molecular mechanism of cadmium (Cd) resistance can provide valuable insights into the tolerance of wheat. Plant proteins known as Topless/Topless–Related (TPL/TPR) play a role in growth, development, defense regulation, and stress response. In this study, we identified TaTPR2 as being induced by Cd stress treatment. Upon Cd treatment, wheat plants overexpressing TaTPR2 exhibited better growth compared to wild–type (WT) plants. Moreover, the transgenic lines showed reduced accumulation of reactive oxygen species (ROS), along with significantly higher activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) compared to WT plants. Additionally, the transgenic lines exhibited lower levels of malondialdehyde (MDA) and electrolyte leakage compared to WT plants. Further analysis revealed that TabHLH41 directly binds to the E–box motif of the TaTPR2 promoter and positively regulates its expression. Overall, the overexpression of TaTPR2 in transgenic wheat resulted in reduced accumulation of Cd and ROS. These findings highlight the significance of the TabHLH41–TaTPR2 pathway as a crucial response to Cd stress in wheat.

The cadCA and cadB/DX operons are possibly induced in cadmium resistance mechanism by Frankia alni ACN14a

BackgroundCadmium (Cd2+) is one of the highly toxic heavy metals and is considered as a carcinogenic agent. Our aim was to confirm the ability of Frankia alni ACN14a to resist Cd2+ and to determine the genes involved in the resistance mechanism. ResultsF. alni ACN14a and Frankia casuarinae CcI3 hyphae showed up to 10 and 22 times Cd2+ accumulation when exposed to 1 mM Cd2+, respectively. Scanning electron microscopy (SEM) exhibited a stable Cd2+ precipitate on the cell surface, and the increase in Cd2+ weight level reached 16.45% when evaluated with SEM-EDAX analysis. The following two potential Cd2+ operons were identified: 1. cadCA operon, which encodes a copper-transporting P-type ATPase A (cadA, FRAAL0989) and an ArsR family regulator (cadC, FRAAL0988), with 37- and 70-fold increase in their expression by qRT-PCR, respectively and 2. cadB/DX, which encodes a putative cobalt-zinc-cadmium resistance protein (cadD, FRAAL3628) and heavy metal-associated domain protein (cadX, FRAAL3626), with 22- and 16-fold upregulation when exposed to Cd2+ stress. ConclusionsCd2+ tolerance by F. alni ACN14a involved efflux of Cd2+ outside the cells and binding it to the membrane surface. Our results indicate the existence of two cadmium-resistance mechanisms in Frankia strains, which support the idea of using them as a bioremediation agent.How to cite: Rehan M, Alhusays A, Serag AM, et al. The cadCA and cadB/DX operons are possibly induced in cadmium resistance mechanism by Frankia alni ACN14a. Electron J Biotechnol 2022;60. https://doi.org/10.1016/j.ejbt.2022.09.006.

Limited Role of Rhamnolipids on Cadmium Resistance for an Endogenous-Secretion Bacterium.

Rhamnolipids, a type of biosurfactant, represent a potential strategy for both enhancing organismic resistance and in situ remediation of heavy metals contaminations. In-depth study of the mechanism of rhamnolipids synthesis in response to heavy metals stress, is indispensable for a wide use of biosurfactant-secreting microbes in bioremediation. In this study, we employed the wild-type and the rhlAB deficient strain (ΔrhlAB) of Pseudomonas aeruginosa, a prototypal rhamnolipids-producing soil microorganism, to investigate its responses to cadmium resistance based on its physicochemical, and physiological properties. Compared with the wild-type strain, the ΔrhlAB were more sensitive to Cd-stress at low Cd concentration (<50 mg/L), whereas there was little difference in sensitivity at higher Cd concentrations, as shown by spot titers and cell viability assays. Secreted rhamnolipids reduced intracellular Cd2+ accumulation to alleviate Cd2+ stress, whereas endogenous rhamnolipids played a limited role in alleviating Cd2+ stress. Synthesized rhamnolipids exhibited a higher critical micelle concentration (CMC) (674.1 mg/L) and lower emulsification index (4.7%) under high Cd-stress, while these parameters showed no obvious changes. High Cd-stress resulted in high hydrophilic wild-type bacterial surface and lower bioremediation ability. This study could advance a deeper understanding of the mechanism of cadmium resistance and provide a theoretical foundation for the application of biosurfactant and biosurfactant-secreted bacterium in contaminant bioremediation.

Resistance properties and adaptation mechanism of cadmium in an enriched strain, Cupriavidus nantongensis X1T

Bacterial adaption to heavy metal stress is a complex and comprehensive process of multi-response regulation. However, the mechanism is largely unexplored. In this study, cadmium (Cd) resistance and adaptation mechanism in Cupriavidus nantongensis X1T were investigated. Strain X1T could resist the stress of 307 mg/L Cd2+ and remove 70% Cd2+ in 48 h. Spectroscopic analyses suggested interactions between Cd2+ with C-N, -COOH, and -NH ligands of extracellular polymeric substances. Whole-genome sequencing found that the resistance of Cd2+ in strain X1T was caused by the joint action of Czc and Cad systems. Cd2+ at 20 mg/L elicited differential expression of 1157 genes in strain X1T. In addition to the reported effects of uptake, adsorption, effluxion, and accumulation system, the oxidative stress system, Type-VI secretory protein system, Fe-S protein synthesis, and cysteine synthesis system in strain X1T were involved in the Cd2+ resistance and accumulation. The intracellular accumulation content of Cd2+ in strain X1T was higher than the extracellular adsorption content made strain X1T to be an important resource strain in the bioremediation of Cd-contaminated sewage. The results provide a theoretical network for understanding the complex regulatory system of bacterial resistance and adaptation of Cd against stressful environments.

Cloning and bioinformatics analysis of cadmium-resistant gene TASFT2 in wheat

Cadmium is a non-essential trace element which is highly toxic to plants. Because of its high mobility and toxicity, it has become a hot topic to study the molecular mechanism of cadmium uptake and transport by plants and to cultivate new crop varieties resistant to cadmium and low cadmium accumulation. Cd enters into the plant body, it will be absorbed by the root system and gradually transported to the above-ground part. Plants reduce toxic effects by absorbing and transporting heavy metals in different chemical forms and storing them in different organs and tissues. Under cadmium stress, plants produce a variety of physiological and biochemical mechanisms that limit cadmium absorption and transfer to reduce cadmium damage. Cadmium stress induces the expression level of metallothionine gene in gramineous crops (wheat and rice), which has a positive effect on improving plant resistance to cadmium and alleviating cadmium toxicity. It is not clear how the gene causes tolerance to heavy metals. Therefore, this experiment cloned the gene and analyzed the biological information to find the mechanism of cadmium resistance.&#x0D; The full length of TaSFT2 gene was cloned by RT-PCR. The sequence analysis showed that ORF gene was 684bp, encoding 228 amino acids, with a molecular weight of 58.542kD and an isoelectric point of 9.16. The results of evolutionary tree analysis showed that wheat TaSFT2 was closely related to maize ZmGot1/Sft2 gene and rice OsGot1/Sft2 protein.&#x0D; The basic information of TaSFT2 gene can be obtained by chromosome location analysis, intron/exon analysis, ORF analysis and expression profile analysis, etc. By analyzing the basic properties of TaSFT2 protein, hydrophobicity analysis, transmembrane region prediction, signal peptide prediction and similarity prediction, the properties of gene-encoded protein can be preliminarily determined and predicted. In particular, hydrophobicity analysis and transmembrane region prediction can be used to predict whether the gene is membrane protein, which has important reference significance for determining the direction of experimental research.

Cadmium resistance mechanisms of a functional strain Enterobacter sp. DNB-S2, isolated from black soil in Northeast China

Enterobacter sp. DNB-S2 is an efficient DBP-degrading bacterial strain, which was proved to be highly resistant to cadmium (MIC was 12.5 mM) and the involved mechanisms were verified in this study. The results showed the maximu values of extracellular adsorbed and intracellular accumulated Cd2+ contents accounted for 24.67% and 4.78% of the initial Cd2+ content in the medium. The extreme low intracellular Cd accumulation reduced the risk of cadmium interfering with normal cell metabolism. TEM exhibited cell structure of DNB-S2 kept intact even at high Cd concentrations. Some precipitation particles were formed inside cells, which were further identified to be Cd–O–P, Cd (OH)2, CdCO3, and CdS by XPS. Furthermore, Cd resistant genes were predicted on DNB-S2 genome, and the presence of 11 alkaline phosphatase encoding genes provided genetic basis for the formation of cadmium phosphate precipitation, which illustrated intracellular precipitation is one of the important intracellular Cd detoxification mechanisms in strain DNB-S2. Noteworthily, 8 efflux proteins were annotated possibly responsible for cadmium efflux and they were all predicted located on cytoplasmic membrane. In a word, integrity of cell structure, the formation of cadmium precipitation and efficient exclusion primarily constitute Cd tolerance mechanisms of strain DNB-S2. This strain exhibits promising application to the bioremediation of cadmium-DBP contaminated bites.

An operon consisting of a P-type ATPase gene and a transcriptional regulator gene responsible for cadmium resistances in Bacillus vietamensis 151\u20136 and Bacillus marisflavi 151\u201325

BackgroundCadmium (Cd) is a severely toxic heavy metal to most microorganisms. Many bacteria have developed Cd2+ resistance.ResultsIn this study, we isolated two different Cd2+ resistance Bacillus sp. strains, Bacillus vietamensis 151–6 and Bacillus marisflavi 151–25, which could be grown in the presence of Cd2+ at concentration up to 0.3 mM and 0.8 mM, respectively. According to the genomic sequencing, transcriptome analysis under cadmium stress, and other related experiments, a gene cluster in plasmid p25 was found to be a major contributor to Cd2+ resistance in B. marisflavi 151–25. The cluster in p25 contained orf4802 and orf4803 which encodes an ATPase transporter and a transcriptional regulator protein, respectively. Although 151–6 has much lower Cd2+ resistance than 151–25, they contained similar gene cluster, but in different locations. A gene cluster on the chromosome containing orf4111, orf4112 and orf4113, which encodes an ATPase transporter, a cadmium efflux system accessory protein and a cadmium resistance protein, respectively, was found to play a major role on the Cd2+ resistance for B. vietamensis 151–6.ConclusionsThis work described cadmium resistance mechanisms in newly isolated Bacillus vietamensis 151–6 and Bacillus marisflavi 151–25. Based on homologies to the cad system (CadA-CadC) in Staphylococcus aureus and analysis of transcriptome under Cd2+ induction, we inferred that the mechanisms of cadmium resistance in B. marisflavi 151–25 was as same as the cad system in S. aureus. Although Bacillus vietamensis 151–6 also had the similar gene cluster to B. marisflavi 151–25 and S. aureus, its transcriptional regulatory mechanism of cadmium resistance was not same. This study explored the cadmium resistance mechanism for B. vietamensis 151–6 and B. marisflavi 151–25 and has expanded our understanding of the biological effects of cadmium.

Improving Cadmium Resistance in Escherichia coli Through Continuous Genome Evolution.

Cadmium (Cd) is a heavy metal that is extremely toxic to many organisms; however, microbes are highly adaptable to extreme conditions, including heavy metal contamination. Bacteria can evolve in the natural environment, generating resistant strains that can be studied to understand heavy-metal resistance mechanisms, but obtaining such adaptive strains usually takes a long time. In this study, the genome replication engineering assisted continuous evolution (GREACE) method was used to accelerate the evolutionary rate of the Escherichia coli genome to screen for E. coli mutants with high resistance to cadmium. As a result, a mutant (8mM-CRAA) with a minimum inhibitory concentration (MIC) of 8 mM cadmium was generated; this MIC value was approximately eightfold higher than that of the E. coli BL21(DE3) wild-type strain. Sequencing revealed 329 single nucleotide polymorphisms (SNPs) in the genome of the E. coli mutant 8mM-CRAA. These SNPs as well as RNA-Seq data on gene expression induced by cadmium were used to analyze the genes related to cadmium resistance. Overexpression, knockout and mutation of the htpX (which encodes an integral membrane heat shock protein) and gor (which encodes glutathione reductase) genes revealed that these two genes contribute positively to cadmium resistance in E. coli. Therefore, in addition to the previously identified cadmium resistance genes zntA and capB, many other genes are also involved in bacterial cadmium resistance. This study assists us in understanding the mechanism of microbial cadmium resistance and facilitating the application of heavy-metal remediation.

Response of the biomining Acidithiobacillus ferrooxidans to high cadmium concentrations

Cadmium is a heavy metal present in contaminated soils. It has no biological role but when entering cells generates DNA damage, overexpression of stress response proteins and misfolded proteins, amongst other deleterious effects. Acidithiobacillus ferrooxidans is an acidophilic bacterium resisting high concentrations of heavy metals such as cadmium. This is important for industrial bioleaching processes where Cd+2 concentrations can be 5–100 mM. Cadmium resistance mechanisms in these microorganisms have not been fully characterized. A. ferrooxidans ATCC 53993 contains genes coding for possible metal resistance determinants such as efflux systems: P-type ATPases, RND transporters and cation diffusion facilitators. In addition, it has extra copies of these genes in its exclusive genomic island (GI). Several of these putative genes were characterized in the present report by determining their transcriptional expression profiles and functionality. Moreover, an iTRAQ proteomic analysis was carried out to explore new cadmium resistance determinants in this bacterium. Changes in iron oxidation components, upregulation of transport proteins and variations in ribosomal protein levels were seen. Finally, increased concentrations of exclusive putative cadmium ATPases present in strain ATCC 53993 GI and other non-identified proteins such as Lferr_0210, forming part of a possible operon, could explain its extreme cadmium resistance. SignificanceCadmium is a very toxic heavy metal present in mining operations and contaminated environments, it can affect all living organisms, including humans. Therefore, it is important to know the resistance mechanisms of bacteria highly resistant to this metal. These microorganisms in turn, can be used to bioremediate more efficiently environments highly polluted with metals. The results obtained suggest A. ferrooxidans strain ATCC 53993 can be an efficient bacterium to remove cadmium, copper and other metals from contaminated sites.

Characterization of a promiscuous cadmium and arsenic resistance mechanism in Thermus thermophilus HB27 and potential application of a novel bioreporter system

BackgroundThe characterization of the molecular determinants of metal resistance has potential biotechnological application in biosensing and bioremediation. In this context, the bacterium Thermus thermophilus HB27 is a metal tolerant thermophile containing a set of genes involved in arsenic resistance which, differently from other microbes, are not organized into a single operon. They encode the proteins: arsenate reductase, TtArsC, arsenic efflux membrane transporter, TtArsX, and transcriptional repressor, TtSmtB.ResultsIn this work we show that the arsenic efflux protein TtArsX and the arsenic responsive transcriptional repressor TtSmtB are required to provide resistance to cadmium. We analyzed the sensitivity to Cd(II) of mutants lacking TtArsX, finding that they are more sensitive to this metal than the wild type strain. In addition, using promoter probe reporter plasmids, we show that the transcription of TtarsX is also stimulated by the presence of Cd(II) in a TtSmtB-dependent way. Actually, a regulatory circuit composed of TtSmtB and a reporter gene expressed from the TtarsX promoter responds to variation in Cd(II), As(III) and As(V) concentrations.ConclusionsOur results demonstrate that the system composed by TtSmtB and TtArsX is responsible for both the arsenic and cadmium resistance in T. thermophilus. The data also support the use of T. thermophilus as a suitable chassis for the design and development of As-Cd biosensors.

PMK-1 p38 MAPK promotes cadmium stress resistance, the expression of SKN-1/Nrf and DAF-16 target genes, and protein biosynthesis in Caenorhabditis elegans

The mechanisms of cadmium (Cd) resistance are complex and not sufficiently understood. The present study, therefore, aimed at assessing the roles of important components of stress-signaling pathways and of ABC transporters under severe Cd stress in Caenorhabditis elegans. Survival assays on mutant and control animals revealed a significant promotion of Cd resistance by the PMK-1 p38 MAP kinase, the transcription factor DAF-16/FoxO, and the ABC transporter MRP-1. Transcriptome profiling by RNA-Seq on wild type and a pmk-1 mutant under control and Cd stress conditions revealed, inter alia, a PMK-1-dependent promotion of gene expression for the translational machinery. PMK-1 also promoted the expression of target genes of the transcription factors SKN-1/Nrf and DAF-16 in Cd-stressed animals, which included genes for molecular chaperones or immune proteins. Gene expression studies by qRT-PCR confirmed the positive effects of PMK-1 on DAF-16 activity under Cd stress and revealed negative effects of DAF-16 on the expression of genes for MRP-1 and DAF-15/raptor. Additional studies on pmk-1 RNAi-treated wild type and mutant strains provided further information on the effects of PMK-1 on SKN-1 and DAF-16, which resulted in a model of these relationships. The results of this study demonstrate a central role of PMK-1 for the processing of cellular responses to abiotic and biotic stressors, with the promoting effects of PMK-1 on Cd resistance mostly mediated by the transcription factors SKN-1 and DAF-16.

Identification of cadmium resistance and adsorption gene from Escherichia coli BL21 (DE3)

Cadmium resistance and adsorption genecapBfromEscherichia coliBL21 (DE3).

Cadmium resistance mechanism in Escherichia coli P4 and its potential use to bioremediate environmental cadmium.

A cadmium-resistant bacterium was isolated from industrial wastewater and identified as Escherichia coli (dubbed as P4) on the basis of morphological, biochemical tests and 16S rRNA ribotyping. It showed optimum growth at 30°C and pH 7. E. coli P4 found to resist Cd(+2) (10.6mM) as well as Zn(+2) (4.4mM), Pb(+2) (17mM), Cu(+2) (3.5mM), Cr(+6) (4.4mM), As(+2) (10.6mM), and Hg(+2) (0.53mM). It could remove 18.8, 37, and 56% Cd(+2) from aqueous medium after 48, 96, and 144h, respectively. Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM), and Energy-dispersive X-ray (EDX) analysis also confirmed the biosorption of Cd(+2) by E. coli P4. However, temperature and pH were found to be the most critical factors in biosorption of Cd(+2) by E. coli P4. Cd(+2) stress altered E. coli P4 cell physiology analyzed by measuring glutathione (GSH) and non-protein thiol (cysteine) levels which were increased up to 130 and 48%, respectively. Quantitative real-time polymerase chain reaction (qRT-PCR) showed alteration in the expression levels of ftsZ, mutS, clpB, ef-tu, and dnaK genes in the presence of Cd(+2). Total protein profiles of E. coli P4 in the absence and presence of Cd(+2) were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), which showed remarkable difference in the banding pattern. czcB gene, a component of czcCBA operon, was amplified from genomic DNA which suggested the chromosomal-borne Cd(+2) resistance in E. coli P4. Furthermore, it harbors smtAB gene which plays a significant role in Cd(+2) resistance.

Decreased accumulation of cadmium in Drosophila selected for resistance suggests a mechanism independent of metallothionein.

Heavy metals, including cadmium, are common contaminants in environments subject to human activity. Responses to exposure in the fruit fly, Drosophila melanogaster, are dosage-dependent and resistance is selectable. While metallothionein-mediated sequestration has been extensively studied as a mechanism of cadmium resistance, a link between selection for resistance and an increased accumulation of cadmium has yet to be demonstrated. To address this need, we have selected wild-type flies for cadmium resistance for 20 generations and tested metal content using mass spectrometry. Resistant flies were observed to contain lower levels of cadmium, arguing for a mechanism of cadmium resistance that is not mediated by increased sequestration. This, coupled with genetic evidence suggesting the involvement of factors located on the X chromosome, suggests a gene other than metallothionein may be involved in resistance in this line.

Cadmium Resistance Mechanism in Acidophilic and Alkalophilic Bacterial Isolates and their Application in Bioremediation of Metal-Contaminated Soil

Acidophilic and alkalophilic cadmium-resistant bacterial strains were isolated from the metal-contaminated soil of Panki thermal power plant, Kanpur, India, and two strains, namely SB21 (acidophilic) and SB20 (alkalophilic), were selected as representative strains on the basis of their high resistance to cadmium as compared to other strains. The phylogenetic analysis of the strains using 16s rDNA sequence revealed the strains to be Pseudomonas putida and Comamonas species strain, respectively. Furthermore, to determine the mechanism involved in cadmium resistance, a czc gene was amplified from the strains and sequenced. Homology of the sequences of the two strains, when compared with the available database using a BLASTn search, showed that the 650bp amplicons possess a partial czcA gene sequence. Moreover, the mechanism was confirmed by the results of atomic absorption spectroscopy and transmission electron microscopy studies, which supported the efflux mechanism in the alkalophilic strain SB20, but the acidophilic strain SB21 showed intracellular and periplasmic accumulation of metal in the cells in spite of the presence of the czcA gene, indicating the presence of multiple mechanisms of metal resistance in strain SB21. Further, the strains were characterized functionally for their bioremediation potential in cadmium-contaminated soil under acidic and alkaline conditions by performing an in-situ greenhouse experiment using mungbean plants. A marked increase in agronomical parameters was observed under acidic conditions in the presence of strain SB21. Moreover, the concentration of metal decreased in both plants and soil in the presence of these bioinoculants; however, strain SB21 was found to be a better decontaminator than SB20 and thus can further be employed in bioremediation applications.

Cd2+ resistance mechanisms in Methanosarcina acetivorans involve the increase in the coenzyme M content and induction of biofilm synthesis

To assess what defence mechanisms are triggered by Cd(2+) stress in Methanosarcina acetivorans, cells were cultured at different cadmium concentrations. In the presence of 100 μM CdCl2, the intracellular contents of cysteine, sulfide and coenzyme M increased, respectively, 8, 27 and 7 times versus control. Cells incubated for 24 h in medium with less cysteine and sulfide removed up to 80% of Cd(2+) added, whereas their cysteine and coenzyme M contents increased 160 and 84 times respectively. Cadmium accumulation (5.2 μmol/10-15 mg protein) resulted in an increase in methane synthesis of 4.5 times in cells grown on acetate. Total phosphate also increased under high (0.5 mM) Cd(2+) stress. On the other hand, cells preadapted to 54 μM CdCl2 and further exposed to > 0.63 mM CdCl2 developed the formation of a biofilm with an extracellular matrix constituted by carbohydrates, DNA and proteins. Biofilm cells were able to synthesize methane. The data suggested that increased intracellular contents of thiol molecules and total phosphate, and biofilm formation, are all involved in the cadmium resistance mechanisms in this marine archaeon.

Metal Accumulation in Cell Wall: A Possible Mechanism of Cadmium Resistance by Pseudomonas stutzeri

A heavy metal resistant strain, Pseudomonas stutzeri (MTCC 101) has been investigated for its cadmium tolerance properties along with its antibiotic resistance. The organism could tolerate cadmium up to 1,200μg/mL with LD50 value 700μg/mL. The gene(s) involved in such high resistance appear(s) to be induced in the presence of the metal. Increasing concentrations of cadmium successively prolonged the lag phase of growth with delayed attainment of the stationary phase. Transmission electron microscope and scanning electron microscope-energy dispersive analysis of X-ray spectroscope analysis showed cadmium adsorption on the bacterial surface with morphological distortion. Atomic absorption spectrometric study corroborated this data, showing highest cadmium accumulation in the cell wall fraction of the bacteria. Additionally, the cell wall fraction showed synthesis of new proteins when grown under metal stress.

Contribution of Cell Walls, Nonprotein Thiols, and Organic Acids to Cadmium Resistance in Two Cabbage Varieties

To study possible cadmium (Cd) resistance mechanisms in cabbage (Brassica oleracea L.), several parameters of metal uptake, distribution, and complexation were compared between two varieties Chunfeng [CF (Cd-tolerant)] and Lvfeng [LF (Cd-sensitive)]. Results showed that CF contained significantly lower Cd concentrations in leaves and higher Cd concentrations in roots than LF. Approximately 70 to 74% and 66 to 68% of Cd taken up by LF and CF, respectively, was transported to shoots. More Cd was bound to the cell walls of leaves, stems, and roots in CF than in LF. The higher capacity of CF to limit Cd uptake into shoots could be explained by immobilization of Cd in root cell walls. Compared with control groups, Cd treatment also significantly increased concentrations of nonprotein thiols, phytochelatins (PCs), and citric acid in the leaves and roots of the two varieties; the increases were more pronounced in CF than in LF. Taken together, the results suggest that the greater Cd resistance in CF than in LF may be attributable to the greater capacity of CF to limit Cd uptake into shoots and complex Cd in cell walls and metal binding ligands, such as PCs and citric acid. However, the contributions of PCs and citric acid to Cd detoxification might be smaller than those in cell walls.