Integrated induction of silver resistance determinants and production of extracellular polymeric substances in Cupriavidus metallidurans BS1 in response to silver ions and silver nanoparticles.

Although the antimicrobial mechanisms of nanomaterials have been extensively investigated, bacterial defense mechanisms associated with AgNPs have not been fully elucidated. We here report that dissolved Ag+ (>0.05 µg mL-1) displayed higher toxicity on cell growth of strain Cupriavidus metallidurans BS1 (GCA_003260185.2) in comparison to 2 and 20 nm AgNPs. The genes necessary for synthesis of distinct abundance and composition of extracellular polymeric substances (EPS) were induced in strain BS1 exposed to Ag stress. This resulted in 20.1% (Ag(I)-EPS) and 24.2% (2 nm AgNPs-EPS) of the CO band integrated intensities being converted into C-OH/C-O-C group vibrations and the Ag-O bond was formed between EPS and 20 nm AgNPs. Meanwhile, the expression of primary resistance genes of the cus, sil and cup operon encoding HME-RND-driven efflux systems as well as a PIB1-type ATPase (CupA) were significantly induced after exposure to Ag(I), 2 and 20 nm AgNPs, respectively. Furthermore, distinct genes involved in biosynthesis pathways responsible for production of EPS were induced to relieve the toxicity of Ag(I), 2 nm and 20 nm AgNPs. This combined action is one potential reason why strain BS1 displayed distinct resistances in response to Ag(I) compared to 2 and 20 nm AgNPs. This work will help in understanding processes important in bacterial defensive mechanisms to AgNPs.

Toxicity of antimony to plants: Effects on metabolism of N and S in a rice plant.

Excess antimony (Sb) has been shown to damage plant growth. Rice plants readily absorb a large amount of Sb after a long period of flooding, yet the mechanisms underlying Sb toxicity in plants have not been solved. This study was conducted to explore the effects of Sb on the uptake of N and S, and monitor the concentrations of reduced glutathione (GSH) and enzymes associated with these processes. In addition, we analyzed differentially expressed metabolites (DEMs) correlated with amino acids (AAs) and oligopeptides, specifically DEMs containing sulfur (S), GSH and indole-3-acetic acid (IAA). The results showed that antimonite [Sb(III)] inhibited shoot growth whereas antimonate [Sb(V)] stimulated shoot growth. Interestingly, Sb(III)5/10 enhanced shoot concentrations of total nitrogen (N), NH4+-N [only at Sb(III)10] and S; but reduced the shoot concentrations of NO3-N and soluble protein. Sb(III)5/10 addition significantly increased oxidized glutathione (GSSG) concentration and activities of glutathione peroxidase (GSH-Px) and glutathione S-transferase (GST) but non-significantly affected concentration of reduced glutathione (GSH) and activities of γ-glutamylcysteine synthetase (GCL) and glutathione reductase (GR), suggesting Sb(III) restricted GSH recycling. Addition of Sb (1) increased the abundance of DEMs associated with lignins, Ca uptake, toxicity/detoxification, and branched chain AAs; (2) decreased the abundance of AAs inclcuding isoleucine (Ile), leucine (Leu), tryptophan (Trp), tyrosine (Tyr) and histidine (His); (3) increased the abundance of arginine (Arg), putrescine (Put) and spermidine (Spd); and (4) affected methylation and acetylation of many AAs, especially acetylation.

A Sb(III)-specific efflux transporter from Ensifer adhaerens E-60.

Antimony is pervasive environmental toxic substance, and numerous genes encoding mechanisms to resist, transform and extrude the toxic metalloid antimony have been discovered in various microorganisms. Here we identified a major facilitator superfamily (MFS) transporter, AntB, on the chromosome of the arsenite-oxidizing bacterium Ensifer adhaerens E-60 that confers resistance to Sb(III) and Sb(V). The antB gene is adjacent to gene encoding a LysR family transcriptional regulator termed LysRars, which is an As(III)/Sb(III)-responsive transcriptional repressor that is predicted to control expression of antB. Similar antB and lysRars genes are found in related arsenic-resistant bacteria, especially strains of Ensifer adhaerens, and the lysRars gene adjacent to antB encodes a member of a divergent subgroup of putative LysR-type regulators. Closely related AntB and LysRars orthologs contain three conserved cysteine residues, which are Cys17, Cys99, and Cys350 in AntB and Cys81, Cys289 and Cys294 in LysRars, respectively. Expression of antB is induced by As(III), Sb(III), Sb(V) and Rox(III) (4-hydroxy-3-nitrophenyl arsenite). Heterologous expression of antB in E. coli AW3110 (Δars) conferred resistance to Sb(III) and Sb(V) and reduced the intracellular concentration of Sb(III). The discovery of the Sb(III) efflux transporter AntB enriches our knowledge of the role of the efflux transporter in the antimony biogeochemical cycle.

Boundary sample-based class-weighted semi-supervised learning for malignant tumor classification of medical imaging.

Medical image classification plays a pivotal role within the field of medicine. Existing models predominantly rely on supervised learning methods, which necessitate large volumes of labeled data for effective training. However, acquiring and annotating medical image data is both an expensive and time-consuming endeavor. In contrast, semi-supervised learning methods offer a promising approach by harnessing limited labeled data alongside abundant unlabeled data to enhance the performance of medical image classification. Nonetheless, current methods often encounter confirmation bias due to noise inherent in self-generated pseudo-labels and the presence of boundary samples from different classes. To overcome these challenges, this study introduces a novel framework known as boundary sample-based class-weighted semi-supervised learning (BSCSSL) for medical image classification. Our method aims to alleviate the impact of intra- and inter-class boundary samples derived from unlabeled data. Specifically, we address reliable confidential data and inter-class boundary samples separately through the utilization of an inter-class boundary sample mining module. Additionally, we implement an intra-class boundary sample weighting mechanism to extract class-aware features specific to intra-class boundary samples. Rather than discarding such intra-class boundary samples outright, our approach acknowledges their intrinsic value despite the difficulty associated with accurate classification, as they contribute significantly to model prediction. Experimental results on widely recognized medical image datasets demonstrate the superiority of our proposed BSCSSL method over existing semi-supervised learning approaches. By enhancing the accuracy and robustness of medical image classification, our BSCSSL approach yields considerable implications for advancing medical diagnosis and future research endeavors.

Selenite reduced cadmium uptake, interfered signal transduction of endogenous phytohormones, and stimulated secretion of tartaric acid based on a combined analysis of non-invasive micro-test technique, transcriptome and metabolome.

Selenium (Se) can reduce uptake and translocation of cadmium (Cd) in plants via plenty of ways, including regulation of root morphology. However, the underlying mechanisms on how Se will regulate root morphology under metal(loid) stresses are not fully illustrated. To fill up this knowledge gap, we investigated the effects of 0.5 mg L-1 selenite (Se(IV)) on root exudates, root morphology, root endogenous hormones, and Cd uptake efficiency of rice under the 1 mg L-1 Cd stress condition. The results showed that Se(IV) significantly reduced shoot and root Cd concentrations, and decreased Cd uptake efficiency via root hairs determined by a non-invasive micro-test (NMT) technology. When compared to the 1 mg L-1 Cd (Cd1) treatment, addition of 0.5 mg L-1 Se(IV) (1) significantly reduced root surface area and tip numbers, and non-significantly reduced root length, but significantly enhanced root diameter and root volume; (2) significantly enhanced concentrations of tartaric acid in the root exudate solution, root auxin (IAA) and root jasmonic acid (JA) via a UHPLC or a HPLC analysis; (3) significantly up-regulated metabolites correlated with synthesis of IAA, JA, gibberellin (GA), and salicylic acid, such as GA53, M-SA, (+/-)7-epi-JA, and derivatives of tryptophan and indole in the metabolome analysis. However, results of transcriptome analysis showed that (1) no upregulated differentially expressed genes (DEGs) were enriched in IAA synthesis; (2) some upregulated DEGs were found to be enriched in JA and GA53 synthesis pathways. In summary, although Se(IV) stimulated the synthesis of IAA, JA, and GA53, it significantly inhibited root growth mainly by 1) affecting signal transduction of IAA and GA; 2) altering IAA polar transport and homeostasis; and 3) regulating DEGs including SAUR32, SAUR36, SAUR76, OsSub33, OsEXPA8, OsEXPA18, and Os6bglu24.

A brief history of metal recruitment in protozoan predation.

Metals and metalloids are used as weapons for predatory feeding by unicellular eukaryotes on prokaryotes. This review emphasizes the role of metal(loid) bioavailability over the course of Earth's history, coupled with eukaryogenesis and the evolution of the mitochondrion to trace the emergence and use of the metal(loid) prey-killing phagosome as a feeding strategy. Members of the genera Acanthamoeba and Dictyostelium use metals such as zinc (Zn) and copper (Cu), and possibly metalloids, to kill their bacterial prey after phagocytosis. We provide a potential timeline on when these capacities first evolved and how they correlate with perceived changes in metal(loid) bioavailability through Earth's history. The origin of phagotrophic eukaryotes must have postdated the Great Oxidation Event (GOE) in agreement with redox-dependent modification of metal(loid) bioavailability for phagotrophic poisoning. However, this predatory mechanism is predicted to have evolved much later - closer to the origin of the multicellular metazoans and the evolutionary development of the immune systems.

Using silkworm excrement to restore vegetation and soil ecology in heavily contaminated mining soils by multiple metal(loid)s: A recyclable sericulture measure.

Ecological restoration of heavily contaminated soils by multiple metal(loid)s in mining areas is very difficult. In this study, we provided an attractive measure of using silkworm excrement (SE) and its modified materials to restore the soil heavily contaminated by arsenic (As), antimony (Sb), cadmium (Cd), lead (Pb) and chromium (Cr). We investigated the adsorption capacities and the associated remediation mechanisms for antimonite [Sb(III)] and antimonate [Sb(V)] by raw SE, biochar-modified SE (BC700), iron-modified BC700 (MBC) and sulfhydryl-modified BC700 (SH). Then, we selected SE and SH to compare their outcomes to restore the vegetations and the soil bacterial communities in the investigated soil mentioned above. The results showed that SE displayed the best characteristics for metal(loid) physical adsorption. But SH conferred the strongest capacity to adsorb Sb (max 23.92 mg g-1), suggesting the process of chemical adsorption played a key role in adsorbing Sb via functional groups (-SH). SE and SH both significantly (1) promoted the growth of pakchoi (Brassica campestris L., New Zealand No.2), community abundance of soil bacteria (283-936 OTUs), and the quantity of bacterial genera correlated with resistance, plant growth promotion and specified carbon metabolism; (2) but reduced bacterial genera correlated with pathogenicity. In this study, we suggested an attractive recyclable measure to restore the disturbed ecological environment in mining areas, i.e, using mulberry to restore the vegetation→ using leaves of mulberry to rear silkworms→ using SE to immobilize metal(loid)s in soils growing mulberry or other plants.

Selenite affected photosynthesis of Oryza sativa L. exposed to antimonite: Electron transfer, carbon fixation, pigment synthesis via a combined analysis of physiology and transcriptome.

Selenium (Se) is a microelement that can counteract (a)biotic stresses in plants. Excess antimony (Sb) will inhibit plant photosynthesis, which can be alleviated by appropriate doses of Se but the associated mechanisms at the molecular levels have not been fully explored. Here, a rice variety (Yongyou 9) was exposed to selenite [Se(IV), 0.2 and 0.8 mg L-1] alone or combined with antimonite [Sb(III), 5 and 10 mg L-1]. When compared to the 10 mg L-1 Sb treatment alone, addition of Se in a dose-dependent manner 1) reduced the heat dissipation efficiency resulting from the inhibited donors, Sb concentrations in shoots and roots, leaf concentrations of fructose, H2O2 and O2•-; 2) enhanced heat dissipation efficiency resulting from the inhibited accepters value, concentrations of Chl a, sucrose and starch, and the enzyme activity of adenosine diphosphate glucose pyrophosphorylase, sucrose phosphate synthase, and sucrose synthase; but 3) did not alter gas exchange parameters, concentrations of Chl b and total Chl, enzyme activity of soluble acid invertase, and values of maximum P700 signal, photochemical efficiency of PSI and electron transport rate of PSI. Se alleviated the damage caused by Sb to the oxygen-evolving complex and promoted the transfer of electrons from QA to QB. When compared to the 10 mg L-1 Sb treatment alone, addition of Se 1) up-regulated genes correlated to synthesis pathways of Chl, carotenoid, sucrose and glucose; 2) disturbed signal transduction pathway of abscisic acid; and 3) upregulated gene expression correlated to photosynthetic complexes (OsFd1, OsFER1 and OsFER2).

Selenite reduced uptake/translocation of cadmium via regulation of assembles and interactions of pectins, hemicelluloses, lignins, callose and Casparian strips in rice roots.

Selenium (Se) can reduce cadmium (Cd) uptake/translocation via regulating pectins, hemicelluloses and lignins of plant root cell walls, but the detailed molecular mechanisms are not clear. In this study, six hydroponic experiments were set up to explore the relationships of uptake/translocation inhibition of Cd by selenite (Se(IV)) with cell wall component (CWC) synthesis and/or interactions. Cd and Se was supplied (alone or combinedly) at 1.0 mg L-1 and 0.5 mg L-1, respectively, with the treatment without Cd and Se as the control. When compared to the Cd1 treatment, the Se0.5Cd1 treatment 1) significantly increased total sugar concentrations in pectins, hemicelluloses and callose, suggesting an enhanced capacity of binding Cd or blocking Cd translocation; 2) stimulated the deposition of Casparian strips (CS) in root endodermis and exodermis to block Cd translocation; 3) stimulated the release of C-O-C (-OH- or -O-) and CO (carboxyl, carbonyl, or amide) to combine Cd; 4) regulated differential expression genes (DEGs) and metabolites (DMs) correlated with synthesis and/or interactions of CWSs to affect cell wall net structure to affect root cell division, subsequent root morphology and finally elemental uptake; and 5) stimulated de-methylesterification of pectins via reducing expression abundances of many DMs and DEGs in the Yang Cycle to reduce supply of methyls to homogalacturonan, and regulated gene expressions of pectin methylesterase to release carboxyls to combine Cd; and 6) down-regulated gene expressions associated with Cd uptake/translocation.

Citrobacter portucalensis Sb-2 contains a metalloid resistance determinant transmitted by Citrobacter phage Chris1.

Bacterial adaptation to extreme environments is often mediated by horizontal gene transfer (HGT) via genetic mobile elements. Nevertheless, phage-mediated HGT conferring bacterial arsenic resistance determinants has rarely been investigated. In this study, a highly arsenite and antimonite resistant bacterium, Citrobacter portucalensis strain Sb-2, was isolated, and genome analysis showed that several putative arsenite and antimonite resistance determinants were flanked or embedded in prophages. Furthermore, an active bacteriophage carrying one of the ars clusters (arsRDABC arsR-yraQ/arsP) was obtained and sequenced. These genes encoding putative arsenic resistance determinants were induced by arsenic and antimony as demonstrated by RT-qPCR, and one gene arsP/yraQ of the ars cluster was shown to give resistance to MAs(III) and Rox(III), thereby showing function. Here, we were able to directly show that these phage-mediated arsenic and antimony resistances play a significant role in adapting to As- and Sb-contaminated environments. In addition, we demonstrate that this phage is responsible for conferring arsenic and antimony resistances to C. portucalensis strain Sb-2.

Toxicity of different forms of antimony to rice plants: Photosynthetic electron transfer, gas exchange, photosynthetic efficiency, and carbon assimilation combined with metabolome analysis.

Antimony (Sb) is a toxic metalloid, and excess Sb causes damage to the plant photosynthetic system. However, the underlying mechanisms of Sb toxicity in the plant photosynthetic system are not clear. Hydroponic culture experiments were conducted to illustrate the toxicity differences of antimonite [Sb(III)] and antimonate [Sb(V)] to the photosynthetic system in a rice plant (Yangdao No. 6). The results showed that Sb(III) showed a higher toxicity than Sb(V), judging from (1) lower shoot and root biomass, leaf water moisture content, water use efficiency, stomatal conductance, net photosynthetic rate, and transpiration rate; (2) higher water vapor deficit, soluble sugar content, starch content, and oligosaccharide content (sucrose, stachyose, and 1-kestose). To further analyze the direction of the photosynthetic products, we conducted a metabonomic analysis. More glycosyls were allocated to the synthesis pathways of oligosaccharides (sucrose, stachyose, and 1-kestose), anthocyanins, salicylic acid, flavones, flavonols, and lignin under Sb stress to quench excess oxygen free radicals (ROS), strengthen the cell wall structure, rebalance the cell membrane, and/or regulate cell permeability. This study provides a complete mechanism to elucidate the toxicity differences of Sb(III) and Sb(V) by exploring their effects on photosynthesis, saccharide synthesis, and the subsequent flow directions of glycosyls.

Fe fortification limits rice Cd accumulation by promoting root cell wall chelation and reducing the mobility of Cd in xylem.

Fe biofortification and Cd mitigation in rice is essential for human health, thus the effects of fertilization with Fe on Cd uptake and distribution in rice need to be comprehensively studied. In this study, we investigated the roles of root iron (Fe)/manganese (Mn) plaques, root cell wall, organic acid, and expressions of Cd-transport related genes in restricting Cd uptake and translocation. The rice plants were exposed to 1 µM CdCl2 with or without the addition of three doses of Fe at 5, 50, and 500 µM EDTA-Na2Fe. The results showed that increasing supply of Fe remarkably reduced Cd accumulation in the shoots, mainly because of inhibited translocation of Cd from roots to shoots. As compared to 5 µM Fe treatment, 500 µM Fe significantly increased the ionic soluble pectin (ISP) content and decreased citric acid (CA) in the roots, thereby providing more Cd-binding sites in the cell wall of roots and reducing the mobility of Cd in xylem. Plant Fe status-mediated CA act as the main chelator for Cd mobilization, rather than through decreasing the pH. However, the plants supplied with low Fe or excess Fe facilitated the uptake of Cd in rice roots, as low Fe up-regulated the expression of Cd-transport related genes and excess Fe enhanced Cd enrichment on the root by iron plaque. Importantly, soil fertilization with Fe strongly reduced Cd accumulation in rice grain. Thus, optimizing the soil environmental Fe could effectively reduce Cd accumulation in the shoots by immobilizing Cd in the roots.

Characterization of Two Highly Arsenic-Resistant Caulobacteraceae Strains of Brevundimonas nasdae: Discovery of a New Arsenic Resistance Determinant.

Arsenic (As), distributed widely in the natural environment, is a toxic substance which can severely impair the normal functions in living cells. Research on the genetic determinants conferring functions in arsenic resistance and metabolism is of great importance for remediating arsenic-contaminated environments. Many organisms, including bacteria, have developed various strategies to tolerate arsenic, by either detoxifying this harmful element or utilizing it for energy generation. More and more new arsenic resistance (ars) determinants have been identified to be conferring resistance to diverse arsenic compounds and encoded in ars operons. There is a hazard in mobilizing arsenic during gold-mining activities due to gold- and arsenic-bearing minerals coexisting. In this study, we isolated 8 gold enrichment strains from the Zijin gold and copper mine (Longyan, Fujian Province, China) wastewater treatment site soil, at an altitude of 192 m. We identified two Brevundimonas nasdae strains, Au-Bre29 and Au-Bre30, among these eight strains, having a high minimum inhibitory concentration (MIC) for As(III). These two strains contained the same ars operons but displayed differences regarding secretion of extra-polymeric substances (EPS) upon arsenite (As(III)) stress. B. nasdae Au-Bre29 contained one extra plasmid but without harboring any additional ars genes compared to B. nasdae Au-Bre30. We optimized the growth conditions for strains Au-Bre29 and Au-Bre30. Au-Bre30 was able to tolerate both a lower pH and slightly higher concentrations of NaCl. We also identified folE, a folate synthesis gene, in the ars operon of these two strains. In most organisms, folate synthesis begins with a FolE (GTP-Cyclohydrolase I)-type enzyme, and the corresponding gene is typically designated folE (in bacteria) or gch1 (in mammals). Heterologous expression of folE, cloned from B. nasdae Au-Bre30, in the arsenic-hypersensitive strain Escherichia coli AW3110, conferred resistance to As(III), arsenate (As(V)), trivalent roxarsone (Rox(III)), pentavalent roxarsone (Rox(V)), trivalent antimonite (Sb(III)), and pentavalent antimonate (Sb(V)), indicating that folate biosynthesis is a target of arsenite toxicity and increased production of folate confers increased resistance to oxyanions. Genes encoding Acr3 and ArsH were shown to confer resistance to As(III), Rox(III), Sb(III), and Sb(V), and ArsH also conferred resistance to As(V). Acr3 did not confer resistance to As(V) and Rox(V), while ArsH did not confer resistance to Rox(V).

As(III) Exposure Induces a Zinc Scarcity Response and Restricts Iron Uptake in High-Level Arsenic-Resistant Paenibacillus taichungensis Strain NC1.

The Gram-positive bacterium Paenibacillus taichungensis NC1 was isolated from the Zijin gold-copper mine and shown to display high resistance to arsenic (MICs of 10 mM for arsenite in minimal medium). Genome sequencing indicated the presence of a number of potential arsenic resistance determinants in NC1. Global transcriptomic analysis under arsenic stress showed that NC1 not only directly upregulated genes in an arsenic resistance operon but also responded to arsenic toxicity by increasing the expression of genes encoding antioxidant functions, such as cat, perR, and gpx. In addition, two highly expressed genes, marR and arsV, encoding a putative flavin-dependent monooxygenase and located adjacent to the ars resistance operon, were highly induced by As(III) exposure and conferred resistance to arsenic and antimony compounds. Interestingly, the zinc scarcity response was induced under exposure to high concentrations of arsenite, and genes responsible for iron uptake were downregulated, possibly to cope with oxidative stress associated with As toxicity. IMPORTANCE Microbes have the ability to adapt and respond to a variety of conditions. To better understand these processes, we isolated the arsenic-resistant Gram-positive bacterium Paenibacillus taichungensis NC1 from a gold-copper mine. The transcriptome responding to arsenite exposure showed induction of not only genes encoding arsenic resistance determinants but also genes involved in the zinc scarcity response. In addition, many genes encoding functions involved in iron uptake were downregulated. These results help to understand how bacteria integrate specific responses to arsenite exposure with broader physiological responses.

Sulfur fertilization integrated with soil redox conditions reduces Cd accumulation in rice through microbial induced Cd immobilization.

Sulfate and water management can be respectively applied to control Cd accumulation in rice, but the interaction mechanisms remain unclear. Three water management coupled with five sulfate application concentrations were employed to investigate rice Cd uptake. Results showed there was a significant interaction between sulfate application and soil redox state, and the highest sulfate treatments reduced rice grain Cd by 63.2, 53.5, and 59.4% under the flooding, flooding-moist alternate (FM), and moist irrigation (M) conditions, respectively. It could be explained by the reduction in rhizosphere soil available Cd and lower transport coefficient from root to aboveground. The Desulfovibrio was demonstrated to participate in CdS precipitation, and its abundance was promoted by sulfate especially under flooding. Additionaly, sulfate application facilitated Cd bounded to FeMn oxides, as rhizosphere soil pH raising under flooding. Under FM and M treatments, sulfate application reduced the abundance of Fe-reducing bacteria Geobacter, and correspondingly reduced Fe and Cd availability in rhizosphere soil. Summarily, Cd transfer from soil to rice can be reduced by applying sulfate fertilizer; which is favored by higher soil moisture because of the higher abundance of Desulfovibrio and lower abundance of Geobacter.

Selenate regulates the activity of cell wall enzymes to influence cell wall component concentration and thereby affects the uptake and translocation of Cd in the roots of Brassica rapa L.

Selenium (Se) can be used to counteract cadmium (Cd) toxicity in plants. However, mechanisms underlying the alleviation of Cd toxicity by Se have not been completely elucidated, especially those by which Se reduces Cd translocation. A hydroponic experiment was performed to illustrate the regulatory mechanisms of Cd transport by selenate (Se (VI)) in pakchoi (Brassica rapa L., LvYou 102). The results showed that this plant had a high accumulation capacity for Cd, and Se(VI) addition restricted Cd translocation from roots to shoots. Se(VI) exposure stimulated the concentrations of pectins and hemicellulose II but reduced the concentration of hemicellulose I in the roots. In many cases, the enzymes pectin methylesterase, polygalacturonase, and ß-galactosidase were dose-dependently triggered by Se(VI) under Cd exposure, but root calcium concentration was significantly lowered (p < 0.05). Xyloglucan endoglycosidase (hydrolase) was triggered by Se(VI) under 2 mg L-1 Cd exposure and cellulase was generally activated by Se(VI) under Cd stress. The above results suggest that Se(VI) up-regulates pectin methylesterase activity, stimulates synthesis of pectins, and down-regulates root Ca concentration to release free carboxyl groups to combine Cd. In this study, the relationships between enzyme activity (e.g., peroxidase, superoxidase and ß-galactosidase), hydrogen peroxide, cell wall structure strengthening/loosening, and Cd toxicity affected by Se(VI) were also discussed.

Identification of a MarR Subfamily That Regulates Arsenic Resistance Genes.

In this study, comprehensive analyses were performed to determine the function of an atypical MarR homolog in Achromobacter sp. strain As-55. Genomic analyses of Achromobacter sp. As-55 showed that this marR is located adjacent to an arsV gene. ArsV is a flavin-dependent monooxygenase that confers resistance to the antibiotic methylarsenite [MAs(III)], the organoarsenic compound roxarsone(III) [Rox(III)], and the inorganic antimonite [Sb(III)]. Similar marR genes are widely distributed in arsenic-resistant bacteria. Phylogenetic analyses showed that these MarRs are found in operons predicted to be involved in resistance to inorganic and organic arsenic species, so the subfamily was named MarRars. MarRars orthologs have three conserved cysteine residues, which are Cys36, Cys37, and Cys157 in Achromobacter sp. As-55, mutation of which compromises the response to MAs(III)/Sb(III). GFP-fluorescent biosensor assays show that AdMarRars (MarR protein of Achromobacter deleyi As-55) responds to trivalent As(III) and Sb(III) but not to pentavalent As(V) or Sb(V). The results of RT-qPCR assays show that arsV is expressed constitutively in a marR deletion mutant, indicating that marR represses transcription of arsV. Moreover, electrophoretic mobility shift assays (EMSAs) demonstrate that AdMarRars binds to the promoters of both marR and arsV in the absence of ligands and that DNA binding is relieved upon binding of As(III) and Sb(III). Our results demonstrate that AdMarRars is a novel As(III)/Sb(III)-responsive transcriptional repressor that controls expression of arsV, which confers resistance to MAs(III), Rox(III), and Sb(III). AdMarRars and its orthologs form a subfamily of MarR proteins that regulate genes conferring resistance to arsenic-containing antibiotics. IMPORTANCE In this study, a MarR family member, AdMarRars was shown to regulate the arsV gene, which confers resistance to arsenic-containing antibiotics. It is a founding member of a distinct subfamily that we refer to as MarRars, regulating genes conferring resistance to arsenic and antimony antibiotic compounds. AdMarRars was shown to be a repressor containing conserved cysteine residues that are required to bind As(III) and Sb(III), leading to a conformational change and subsequent derepression. Here we show that members of the MarR family are involved in regulating arsenic-containing compounds.

Application of inorganic selenium to reduce accumulation and toxicity of heavy metals (metalloids) in plants: The main mechanisms, concerns, and risks.

Anthropogenic activities such as mining, industrialization and subsequent emission of industrial waste, and agricultural practices have led to an increase in the accumulation of metal(loid)s in agricultural soils and crops, which threatens the health of people; the risk is more pronounced for individuals whose survival depends on food sources from several contaminated regions. Selenium (Se) is an element essential for the normal functioning of the human body and is a beneficial element for plants. Se deficiency in the diet is a common issue in many countries around the world, such as China and Egypt. >40 diseases are associated with Se deficiency. In practice, Se compounds have been applied through foliar sprays or via base application of fertilizers to increase Se concentration in the edible parts of crops and to satisfy the daily Se intake. Moreover, Se at low concentrations has been used to mitigate the toxicity of many metal(loid)s. In this review, we present an overview of the latest knowledge and practices with regards to the utilization of Se to reduce the uptake/toxicity of metal(loid)s in plants. We have focused on the following issues: 1) the current status of understanding the mechanisms of detoxification and uptake restriction of metal(loid)s regulated by Se; 2) the optimal dose and speciation of Se, and stage of plant growth that is optimal for application; 3) the differences in the efficiency of different application methods of Se including seed priming, base application, and foliar spray of Se fertilizers; 4) the possibility of using Se along with other methods to reduce multiple metal(loid) accumulation in crops; and 5) potential risks when Se is used to reduce metal(loid) accumulation in crops.

Potential of cadmium resistant Burkholderia contaminans strain ZCC in promoting growth of soy beans in the presence of cadmium.

Bioremediation of Cd contaminated environments can be assisted by plant-growth-promoting bacteria (PGPB) enabling plant growth in these sites. Here a gram-negative Burkholderia contaminans ZCC was isolated from mining soil at a copper-gold mine. When exposed to Cd(II), ZCC displayed high Cd resistance and the minimal inhibitory concentration was 7 mM in LB medium. Complete genome analysis uncovered B. contaminans ZCC contained 3 chromosomes and 2 plasmids. One of these plasmids was shown to contain a multitude of heavy metal resistance determinants including genes encoding a putative Cd-translocating PIB-type ATPase and an RND-type related to the Czc-system. These additional heavy metal resistance determinants are likely responsible for the increased resistance to Cd(II) and other heavy metals in comparison to other strains of B. contaminans. B. contaminans ZCC also displayed PGPB traits such as 1-aminocyclopropane-1-carboxylate deaminase activity, siderophore production, organic and inorganic phosphate solubilization and indole acetic acid production. Moreover, the properties and Cd(II) binding characteristics of extracellular polymeric substances was investigated. ZCC was able to induce extracellular polymeric substances production in response to Cd and was shown to be chemically coordinated to Cd(II). It could promote the growth of soybean in the presence of elevated concentrations of Cd(II). This work will help to better understand processes important in bioremediation of Cd-contaminated environment.

Co-selection of antibiotic resistance genes, and mobile genetic elements in the presence of heavy metals in poultry farm environments.

Environmental selection of antibiotic resistance genes (ARGs) is considered to be caused by antibiotic or metal residues, frequently used in livestock. In this study we examined three commercial poultry farms to correlate the co-occurrence patterns of antibiotic and metal residues to the presence of ARGs. We quantified 283 ARGs, 12 mobile genetic elements (MGEs), 49 targeted antibiotics, 7 heavy metals and sequenced 16S rRNA genes. The abundance and type of ARG were significantly enriched in manure while soil harbored the most diverse bacterial community. Procrustes analysis displayed significant correlations between ARGs/MGEs and the microbiome. Cadmium (Cd), arsenic (As), zinc (Zn), copper (Cu) and lead (Pb) were responsible for a majority of positive correlations to ARGs when compared to antibiotics. Integrons and transposons co-occurred with ARGs corresponding to 9 classes of antibiotics, especially Class1 integrase intI-1LC. Redundancy analysis (RDA) and Variance partitioning analysis (VPA) showed that antibiotics, metals, MGEs and bacteria explain solely 0.7%, 5.7%, 12.4%, and 21.9% of variances of ARGs in the microbial community, respectively. These results suggested that bacterial composition and horizontal gene transfer were the major factors shaping the composition of ARGs; Metals had a bigger effect on ARG profile than detected antibiotics in this study.