Zn Sequestration Research Articles

Fabrication of mango-leaf biowaste mediated green magnetite (Fe3O4@MBE NPs) and modified EDTA/Fe3O4@MBE NCs for enhanced heavy metals sequestration: Characterisation, simulation modelling, mechanism, cost-effectiveness and electroplating wastewater performance

This work reports biogenic green fabricated novel magnetite nanoparticles (Fe3O4@MBE NPs) coated with mango-leaf biowaste extract and modified disodium ethylenediamine tetra-acetate (EDTA) functionalised nanocomposite (EDTA/Fe3O4@MBE NCs). Several approaches were used to characterise them, and their applicability as nanoadsorbents was exhaustively investigated in batch mode sequestration of Cd(II), Ni(II), and Zn(II) from synthetic solutions. The Brunauer Emmett and Teller’s surface areas of Fe3O4@MBE and EDTA/Fe3O4@MBE was determined to be 47.73 and 43.95 m2/g, respectively, with average porous diameters of 10.36 and 7.44 nm, revealing mesoporosity. EDTA/Fe3O4@MBE NCs exhibit soft ferromagnetism with a saturation magnetisation of 40.67 emu/g. Further, EDTA modification creates carboxyl and amino sites, improving adsorption performance. Adsorption was pH dependent, with optimal sequestration at 40 mg/L divalent metal ions, 0.5 g/L nanoadsorbent dosage, and 75 min of reaction duration at 27 °C. Among the simulation models employed, the Langmuir isotherm, pseudo second order kinetic, and Elovich kinetic were the best fitted and offered the highest capability to explain the adsorption process. The Langmuir monolayer adsorption capacity (qmax) of 147.06, 129.87, and 117.65 mg/g for Cd(II), Ni(II), and Zn(II) by EDTA/Fe3O4@MBE, respectively, were significantly higher than Fe3O4@MBE (120.48, 102.04, and 95.24 mg/g). The best-suited kinetics precisely describe chemisorption as the primary rate-limiting phase in divalent metal ions adsorption. Thermodynamics approved the endothermic nature and feasibility of divalent metals adsorption, with modest improvement in efficacy at higher temperatures. The ability of EDTA/Fe3O4@MBE to form stable surface complexes with divalent metals enhances adsorption even in the presence of other co-existing ionic species. EDTA/Fe3O4@MBE NCs magnetic separability, improved sequestration upto five recycles, and regenerability of ≥89.07 % demonstrate its cost-effectiveness in heavy metals sequestration. Finally, practicality study confirmed that EDTA/Fe3O4@MBE nanoadsorbent provides a practical and economically efficient option for heavy metals sequestration from electroplating wastewater; thus, it contributed towards UN-2030, SDG No-6.

Root Zn sequestration transporter heavy metal ATPase 3 from Odontarrhena chalcidica enhance Cd tolerance and accumulation in Arabidopsis thaliana

The Ni hyperaccumulator Odontarrhena chalcidica (formerly Alyssum murale), exhibits a significant capacity to accumulate Zn in the roots. However, the molecular mechanisms underlying the variation in Ni and Zn accumulation are poorly understood. Here, we isolated a homolog of heavy metal ATPase 3 from O. chalcidica (OcHMA3) and characterized its functions using heterologous systems. Phylogenetic analysis revealed that OcHMA3 protein shares 87.6 % identity with AtHMA3, with similar metal binding sites to other HMA3 proteins. Heterologous expression of OcHMA3 in yeast increased sensitivity to Cd, Ni and Zn, suggesting it functions as a broad-specificity transporter. Further investigation showed OcHMA3 is constitutively expressed in the roots and localized to the tonoplast. Overexpression of OcHMA3 in A. thaliana shoots increased its roots Zn concentrations by 41.9 % – 74.1 %. However, overexpression of OcHMA3 in roots enhanced its tolerance to Cd and increased roots Cd concentrations by 50.9 % – 90.6 %. Our findings indicated OcHMA3 is responsible for Zn sequestration in root vacuoles, likely leading to Zn retention in roots and subsequent Ni hyperaccumulation in shoots. This study elucidates the molecular mechanism of Ni and Zn accumulation in O. chalcidica, and identifies OcHMA3 as a potential gene for developing Zn-rich plants and for phytoextraction in Cd-contaminated soils.

Molecular modeling of multi-target analogs of huperzine A and applications in Alzheimer's disease.

Given the diverse pathophysiological mechanisms underlying Alzheimer's disease, it is improbable that a single targeted drug will prove successful as a therapeutic strategy. Therefore, exploring various hypotheses in drug design is imperative. The sequestration of Fe(II) and Zn(II) cations stands out as a crucial mechanism based on the mitigation of reactive oxygen species. Moreover, inhibiting acetylcholinesterase represents a pivotal strategy to enhance acetylcholine levels in the synaptic cleft. This research aims to investigate the analogs of Huperzine A, documented in scientific literature, considering of these two hypotheses. Consequently, the speciation chemistry of these structures with Fe(II) and Zn(II) was scrutinized using quantum chemistry calculations, molecular docking simulations, and theoretical predictions of pharmacokinetics properties. From the pharmacokinetic properties, only two analogs, HupA-A1 and HupA-A2, exhibited a theoretical permeability across the blood-brain barrier; on the other hand, from a thermodynamic standpoint, the enantiomers of HupA-A2 showed negligible chelation values. The enantiomers with the most favorable interaction parameters were S'R'HupA-A1 (ΔGBIND = -40.0 kcal mol-1, fitness score = 35.5) and R'R'HupA-A1 (ΔGBIND = -35.5 kcal mol-1, fitness score = 22.61), being compared with HupA (ΔGBIND = -41.75 kcal mol-1, fitness score = 39.95). From this study, some prime candidates for promising drug were S'R'HupA-A1 and R'R'HupA-A1, primarily owing to their favorable thermodynamic chelating capability and potential anticholinesterase mechanism. Quantum chemistry calculations were carried out at B3LYP/6-31G(d) level, considering the IEF-PCM(UFF) implicit solvent model for water. The coordination compounds were assessed using the Gibbs free energy variation and hard and soft acid theory. Molecular docking calculations were conducted using the GOLD program, based on the crystal structure of the acetylcholinesterase protein (PDB code = 4EY5), where the ChemScore function was employed with the active site defined as the region within a 15-Å radius around the centroid coordinates (X = -9.557583, Y = -43.910473, Z = 31.466687). Pharmacokinetic properties were predicted using SwissADME, focusing on Lipinski's rule of five.

Sequestration of Ni (II), Pb (II), and Zn (II) utilizing biogenic synthesized Fe3O4/CLPC NCs and modified Fe3O4/CLPC@CS NCs: Process optimization, simulation modeling, and feasibility study.

The present study reports low-cost novel biogenic magnetite Citrus limetta peels carbon (Fe3O4/CLPC) nanocomposites and modified Fe3O4/CLPC@CS nanocomposites cross-linked with glutaraldehyde and subsequently employed in batch mode sequestration of heavy metals ions. Diverse techniques fully characterized them, and the influence of operating variables on adsorption reactions from aqueous solutions was investigated. The Brunauer, Emmett, and Teller (BET) surface areas of synthesized Fe3O4/CLPC and Fe3O4/CLPC@CS NCs were 53.91 and 32.16 m2/g, while the mesoporous diameters were 7.69 and 7.57nm, respectively. The Langmuir isotherm and Pseudo second order kinetic were well-fitting and capable of explaining the adsorption reaction. The Langmuir-based monolayer adsorption (qmax) for Fe3O4/CLPC@CS NCs was 82.65, 95.24, and 64.10mg/g, higher than Fe3O4/CLPC NCs, which were 70.92, 84.75, and 59.17mg/g for Ni (II), Pb (II), and Zn (II), respectively. Each metal's pseudo second order correlation coefficient (R2 ≥ 0.99) reveals that nanocomposites surface binding functional groups controlled the adsorption rate via chemisorption. Further, thermodynamic results confirm that each studiedmetal ions' adsorption was spontaneous, endothermic, and characterized by an increase in randomness. In addition to magnetic separability, three ad-desorption cycles yielded exceptional adsorption efficacy and > 93% regenerability. The present study also reveals the effective utilization of Fe3O4/CLPC and Fe3O4/CLPC@CS NCs as cost-effective magnetic separable green adsorbents for heavy metals sequestration from electroplating wastewater.

Differential Effects of Senescence on the Phloem Exports of Cadmium and Zinc from Leaves to Grains in Rice during Grain Filling.

In rice, non-essential toxic cadmium (Cd) and the essential nutrient zinc (Zn) share similar transport pathways, which makes it challenging to differentially regulate the allocation of these elements to the grain. The phloem is the main pathway for the loading of these elements into rice grains. It has long been accepted that tissue senescence makes the nutrients (e.g., Zn) stored in leaves available for further phloem export toward the grain. Whether senescence could drive the phloem export of Cd remains unclear. To this end, the stable isotopes 111Cd and 67Zn were used to trace the phloem export and the subsequent allocation of Cd and Zn from the flag leaves, where senescence was accelerated by spraying abscisic acid. Furthermore, changes upon senescence in the distribution of these elements among the leaf subcellular fractions and in the expression of key transporter genes were investigated. Abscisic acid-induced senescence enhanced the phloem export of Zn but had no impact on that of Cd, which was explained by the significant release of Zn from the chloroplast and cytosol fractions (concentrations decreased by ~50%) but a strong allocation of Cd to the cell wall fraction (concentration increased by ~90%) during senescence. Nevertheless, neither Zn nor Cd concentrations in the grain were affected, since senescence strengthened the sequestration of phloem-exported Zn in the uppermost node, but did not impact that of phloem-exported Cd. This study suggests that the agronomic strategies affecting tissue senescence could be utilized to differentially regulate Cd and Zn allocation in rice during grain filling.

Localized Infections with P. aeruginosa Strains Defective in Zinc Uptake Reveal That Zebrafish Embryos Recapitulate Nutritional Immunity Responses of Higher Eukaryotes.

The innate immune responses of mammals to microbial infections include strategies based on manipulating the local concentration of metals such as iron (Fe) and zinc (Zn), commonly described as nutritional immunity. To evaluate whether these strategies are also present in zebrafish embryos, we have conducted a series of heart cavity-localized infection experiments with Pseudomonas aeruginosa strains characterized by a different ability to acquire Zn. We have found that, 48 h after infection, the bacterial strains lacking critical components of the Zn importers ZnuABC and ZrmABCD have a reduced colonization capacity compared to the wild-type strain. This observation, together with the finding of a high level of expression of Zur-regulated genes, suggests the existence of antimicrobial mechanisms based on Zn sequestration. However, we have observed that strains lacking such Zn importers have a selective advantage over the wild-type strain in the early stages of infection. Analysis of the expression of the gene that encodes for a Zn efflux pump has revealed that at short times after infection, P. aeruginosa is exposed to high concentrations of Zn. At the same time, zebrafish respond to the infection by activating the expression of the Zn transporters Slc30a1 and Slc30a4, whose mammalian homologs mediate a redistribution of Zn in phagocytes aimed at intoxicating bacteria with a metal excess. These observations indicate that teleosts share similar nutritional immunity mechanisms with higher vertebrates, and confirm the usefulness of the zebrafish model for studying host-pathogen interactions.

Spatial distribution and sources of heavy metals in the sediment and soils of the Yancheng coastal ecosystem and associated ecological risks.

Studies of heavy metal pollution are essential for the protection of coastal environments. In this study, positive matrix factorization (PMF) and a GeoDetector model were used to evaluate the sources of heavy metal contamination and associated ecological risks along the Yancheng Coastal Wetland. The distribution of heavy metals was shown to be greatly affected by clay content, except for Cr in shoal. Components from 6.5 to 9φ have the strongest ability to absorb heavy metals, where the effects of Cd and Zn sequestration in the wetlands were most apparent. The abilities of various wetland environments to sequester heavy metals were shown to be Spartina alterniflora wetland > woodland > Phragmites australis wetland > aquaculture pond > shoal > paddy > meadow > dry land. The sources of the heavy metals included parent soil material (59%), agriculture (15%), and industrial pollutants (26%). According to the single-factor pollution index, there was no evidence of pollution except Cr and Pb. In general, the heavy metal pollution was insignificant. The order of pollution loading index was shoal > paddy field > dry land > Spartina Alterniflora wetland > aquaculture ponds > woodland > meadow > Phragmites australis wetland. The ecological harm of heavy metal exposure was slight except for Cd and Hg, where vehicle emissions appeared to be the main cause of heavy metal pollution.

Enhanced Zn(II) adsorption by chemically modified sawdust based biosorbents.

Heavy metals present in industrial effluents, when discharged into water channels, not only affect humans but also negatively impact plants and aquatic organisms. Sawdust is available readily in developing countries and can be used by small-scale industries for effluent water treatment containing low concentrations of bivalent zinc ions. This study explores the potential of sawdust-derived biosorbents, after boiling (SDB), chemical modification with formaldehyde (SDF), and sulfuric acid (SDS), for sequestration of Zn(II) from simulated wastewater as well as industrial effluents. The morphological analysis of the three biosorbents indicated a suitable porous structure with a pore size of 232.928 m2/g (SDB), 291.102 m2/g (SDF), and 498.873 m2/g (SDS). The functional analysis of native and metal-laden biosorbents indicated the role of - OH, - C = O, and - NH functional groups in Zn(II) binding. The process parameters were optimized and the spontaneous adsorption of Zn(II) was found to proceed by multilayer formation by following pseudo-second-order kinetics. SDS adsorbent (0.1g) exhibited a greater potential for removal of Zn(II) from industrial effluents as compared to SDB and SDF at pH = 6.0 with the equilibrium adsorption capacity of 45.87mg/g. Therefore, SDS could be a promising adsorbent for the treatment of wastewater in small-scale industries.

Dynamic regulation of Zn(II) sequestration by calgranulin C.

Calgranulin C performs antimicrobial activity in the human immune response by sequestering Zn(II). This biological function is afforded with the aid of two structurally distinct Ca(II)-binding EF hand motifs, wherein one of which bears an unusual amino acid sequence. Here, we utilize solution state NMR relaxation measurements to investigate the mechanism of Ca(II)-modulated enhancement of Zn(II) sequestration by calgranulin C. Using C13 /N15 CPMG dispersion experiments we have measured pH-dependent major and minor state populations exchanging on micro-to-millisecond timescale. This conformational exchange takes place exclusively in the Ca(II)-bound state and can be mapped to residues located in the EF-I loop and the linker between the tandem EF hands. Molecular dynamics (MD) simulations spanning nano-to-microsecond timescale offer insights into the role of pH-dependent electrostatic interactions in EF-hand dynamics. Our results suggest a pH-regulated dynamic equilibrium of conformations that explore a range of "closed" and partially "open" sidechain configurations within the Zn(II) binding site. We propose a novel mechanism by which Ca(II) binding to a non-canonical EF loop regulates its flexibility and tunes the antimicrobial activity of calgranulin C.

S100A12 promotes Mn(II) binding to pneumococcal PsaA and staphylococcal MntC by Zn(II) sequestration

Human S100A12 (calgranulin C, EN-RAGE) is a Zn(II)-sequestering host-defense protein that contributes to the metal-withholding innate immune response against microbial pathogens. S100A12 coordinates Zn(II) ions at two His3Asp sites with high affinity. A similar His3Asp site found in calprotectin (S100A8/S100A9, calgranulin A/B), a closely related human S100 protein, can sequester divalent metal ions from the solute-binding proteins (SBPs) pneumococcal PsaA (pneumococcal surface protein A) and staphylococcal MntC (manganese transport protein C). Both SBPs are components of Mn(II) transporters and capture extracellular Mn(II) ions for subsequent delivery into the bacterial cytosol. Nevertheless, PsaA and MntC exhibit a thermodynamic preference for Zn(II) over Mn(II), and Zn(II) binding can interfere with Mn(II) acquisition. In this work, we have used a biotinylated variant of S100A12 to show that S100A12 can sequester Zn(II) ions from PsaA and MntC. Moreover, electron paramagnetic resonance (EPR) spectroscopy indicates that by sequestering Zn(II) from Zn(II)-bound PsaA and MntC, S100A12 promotes Mn(II) binding to the SBPs. These results inform the function of S100A12 in Zn(II) sequestration, and further suggest that Zn(II)-sequestering S100 proteins may inadvertently protect bacterial pathogens during infection.

AtTIP2;2 facilitates resistance to zinc toxicity via promoting zinc immobilization in the root and limiting root-to-shoot zinc translocation in Arabidopsis thaliana

Zinc (Zn) is an essential micronutrient for plants. However, excess Zn is toxic to non-accumulating plants like Arabidopsis thaliana. To cope with Zn toxicity, non-accumulating plants need to keep excess Zn in the less sensitive root tissues and restrict its translocation to the vulnerable shoot tissues, a process referred to as Zn immobilization in the root. However, the mechanism underlying Zn immobilization is not fully understood. In Arabidopsis, sequestration of excess Zn to the vacuole of root cells is crucial for Zn immobilization, facilitated by distinct tonoplast-localized transporters. As some members of the aquaporin superfamily have been implicated in transporting metal ions besides polar but non-charged small molecules, we tested whether Arabidopsis thaliana tonoplast intrinsic proteins (AtTIPs) could be involved in Zn immobilization and resistance. We found that AtTIP2;2 is involved in retaining excess Zn in the root, limiting its translocation to the shoot, and facilitating its accumulation in the leaf trichome. Furthermore, when expressed in yeast, the tonoplast-localized AtTIP2;2 renders glutathione (GSH)-dependent Zn resistance to yeast cells, suggesting that AtTIP2;2 facilitates the across-tonoplast transport of GSH-Zn complexes. Our findings provide new insights into aquaporins' roles in heavy metal resistance and detoxification in plants.

Zinc transport in rice: how to balance optimal plant requirements and human nutrition.

Zinc (Zn) is an essential micronutrient for both plants and animals, while its deficiency in crops and humans is a global problem that affects both crop productivity and human health. Since plants and humans differ in their Zn requirements, it is crucial to balance plant nutrition and human nutrition for Zn. In this review, we focus on the transport system of Zn from soil to grain in rice (Oryza sativa), which is a major dietary source of Zn for people subsiding on rice-based diets. We describe transporters belonging to the different families that are involved in the uptake, vacuolar sequestration, root-to-shoot translocation, and distribution of Zn, and discuss their mechanisms of regulation. We give examples for enhancing Zn accumulation and bioavailability in rice grains through the manipulation of genes that are highly expressed in the nodes, where Zn is deposited at high concentrations. Finally, we provide our perspectives on breeding rice cultivars with both increased tolerance to Zn-deficiency stress and high Zn density in the grains.

Transcriptional Regulation of Genes Involved in Zinc Uptake, Sequestration and Redistribution Following Foliar Zinc Application to Medicago sativa.

Zinc (Zn) is an essential micronutrient for plants and animals, and Zn deficiency is a widespread problem for agricultural production. Although many studies have been performed on biofortification of staple crops with Zn, few studies have focused on forages. Here, the molecular mechanisms of Zn transport in alfalfa (Medicago sativa L.) were investigated following foliar Zn applications. Zinc uptake and redistribution between shoot and root were determined following application of six Zn doses to leaves. Twelve putative genes encoding proteins involved in Zn transport (MsZIP1-7, MsZIF1, MsMTP1, MsYSL1, MsHMA4, and MsNAS1) were identified and changes in their expression following Zn application were quantified using newly designed RT-qPCR assays. These assays are the first designed specifically for alfalfa and resulted in being more efficient than the ones already available for Medicago truncatula (i.e., MtZIP1-7 and MtMTP1). Shoot and root Zn concentration was increased following foliar Zn applications ≥ 0.1 mg plant−1. Increased expression of MsZIP2, MsHMA4, and MsNAS1 in shoots, and of MsZIP2 and MsHMA4 in roots was observed with the largest Zn dose (10 mg Zn plant−1). By contrast, MsZIP3 was downregulated in shoots at Zn doses ≥ 0.1 mg plant−1. Three functional gene modules, involved in Zn uptake by cells, vacuolar Zn sequestration, and Zn redistribution within the plant, were identified. These results will inform genetic engineering strategies aimed at increasing the efficiency of crop Zn biofortification.

Expression of a Brassica napus metal transport protein (BnMTP3) in Arabidopsis thaliana confers tolerance to Zn and Mn

The essential micronutrient elements zinc (Zn) and manganese (Mn) are crucial for plant growth and development. As an important oil crop, the yield and quality of rapeseed are affected by Zn and Mn toxicity. The cation diffusion facilitator (CDF) family of proteins play significant roles in maintaining intracellular ionic homeostasis and tolerance in plants. However, research on CDF proteins in rapeseed is lacking. In this study, the function of a Brassica napus cation diffusion facilitator/ metal tolerance protein (CDF/MTP) was investigated. The protein, abbreviated BnMTP3 is homologous to the Arabidopsis thaliana MTP3 (AtMTP3). Heterologous expression of BnMTP3 in yeast enhanced tolerance and intracellular sequestration of Zn and Mn. Expression of BnMTP3 in A. thaliana increased Zn and Mn tolerance and markedly increased Zn accumulation in roots. Quantitative RT-PCR analysis showed that BnMTP3 is primarily expressed in roots, and subcellular localization suggested that BnMTP3 is localized in the trans-Golgi network (TGN) and the prevacuolar compartment (PVC) in Arabidopsis and rape protoplast. After treatment with Zn and Mn, BnMTP3 was observed on the vacuolar membrane in transgenic Arabidopsis lines. These findings suggest that BnMTP3 confers Zn and Mn tolerance by sequestering Zn and/or Mn into the vacuole.

Nanoscale trace metal imprinting of biocalcification of planktic foraminifers by Toba\u2019s super-eruption

Bioactive metal releases in ocean surface water, such as those by ash falls during volcanic super-eruptions, might have a potentially toxic impact on biocalcifier planktic microorganisms. Nano-XRF imaging with the cutting-edge synchrotron hard X-ray nano-analysis ID16B beamline (ESRF) revealed for the first time a specific Zn- and Mn-rich banding pattern in the test walls of Globorotalia menardii planktic foraminifers extracted from the Young Toba Tuff layer, and thus contemporaneous with Toba’s super-eruption, 74,000 years ago. The intra-test correlation of Zn and Mn patterns at the nanoscale with the layered calcareous microarchitecture, indicates that the incorporation of these metals is syngenetic to the wall growth. The preferential Mn and Zn sequestration within the incipient stages of chamber formation suggests a selective incorporation mechanism providing a resilience strategy to metal pollution in the test building of planktic foraminifers.

Type I Interferons Ameliorate Zinc Intoxication of Candida glabrata by Macrophages and Promote Fungal Immune Evasion.

SummaryHost and fungal pathogens compete for metal ion acquisition during infectious processes, but molecular mechanisms remain largely unknown. Here, we show that type I interferons (IFNs-I) dysregulate zinc homeostasis in macrophages, which employ metallothionein-mediated zinc intoxication of pathogens as fungicidal response. However, Candida glabrata can escape immune surveillance by sequestering zinc into vacuoles. Interestingly, zinc-loading is inhibited by IFNs-I, because a Janus kinase 1 (JAK1)-dependent suppression of zinc homeostasis affects zinc distribution in macrophages as well as generation of reactive oxygen species (ROS). In addition, systemic fungal infections elicit IFN-I responses that suppress splenic zinc homeostasis, thereby altering macrophage zinc pools that otherwise exert fungicidal actions. Thus, IFN-I signaling inadvertently increases fungal fitness both in vitro and in vivo during fungal infections. Our data reveal an as yet unrecognized role for zinc intoxication in antifungal immunity and suggest that interfering with host zinc homeostasis may offer therapeutic options to treat invasive fungal infections.

Burkholderia pyrrocinia strain JK-SH007 affects zinc (Zn) accumulation and translocation in tomato

ABSTRACT Zinc (Zn) contamination in soil has been a serious problem world widely. Plant growth-promoting bacterium (PGPB) could have potential applications in Zn bioremediation. In this study, Burkholderia pyrrocinia strain JK-SH007 had a high tolerance to Zn and was able to produce indole acetic acid and siderophore, to solubilize inorganic phosphate. And it was found to promote tomato growth and tolerate to high concentration of Zn. We employed a novel combination of transmission electron microscopy (TEM) and scanning electron microscopy with electron dispersive X-ray analysis (SEM-EDX) for understanding the mechanism of Zn toxicity on plant tip morphology, ultrastructure and bacterium protection tomato cell. The high Zn soil supplement was highly toxic to the plants, biomass decreased, root growth inhibited, shoots rotted and chlorisis induced. ‘JK-SH007ʹ improved the relative tolerance of tomato to Zn, enhanced Zn sequestration, alleviated the associated plant damage relative to PGPB-free controls. ‘JK-SH007ʹ may be used to assist metal-hyperaccumulating plants in bioremediation.

Correlation of the expression of metal tolerance protein genes with zinc accumulation and distribution in Malus halliana seedlings under zinc toxicity stress

The micronutrient zinc plays an essential role in the physiological and metabolic processes in plants; however, it becomes very toxic when present in elevated amounts. To evaluate the correlation of metal tolerance proteins (MTP) with Zn accumulation and distribution in Malus halliana seedlings under zinc toxicity stress, seedlings with 8 true leaves were exposed to different Zn concentrations (4, 25, 50, 100 μmol·L−1) in a hydroponic culture system. We found that excessive Zn stress leads to a decrease in plant growth indexes and the accumulation and distribution of Zn in Malus halliana. With increasing levels of Zn stress, the Zn concentration in roots was significantly higher than in stems and leaves. At 100 μmol·L−1 Zn, the Zn concentration in roots, stems and leaves increased by 45.88, 26.51 and 25.43-fold when compare to CK, respectively. The increase of bioaccumulation factor (BAF) and the decrease of translocation factor (TF) all indicated that Zn was accumulated in the roots. Correlation and hierarchical clustering analysis found that the expression of MTP genes is closely associated with Zn concentrations. The MTP2, MTP11 and MTPC2-like genes were significantly negatively correlated with Zn concentration in roots and were resistant to excessive Zn concentration and sequestration. The expression of MTP1 had organ-specificity that was significantly higher in stems and leaves than that in roots. The expression of MTPC4-like was significantly positively correlated with Zn concentration under the treatment of 4 μmol·L−1 Zn, but the correlation decreased under the high concentration of Zn stress, which indicated that the expression of MTP gene was affected by Zn concentration.

The Immune Protein Calprotectin Impacts Clostridioides difficile Metabolism through Zinc Limitation.

The intestines house a diverse microbiota that must compete for nutrients to survive, but the specific limiting nutrients that control pathogen colonization are not clearly defined. Clostridioides difficile colonization typically requires prior disruption of the microbiota, suggesting that outcompeting commensals for resources is critical to establishing C. difficile infection (CDI). The immune protein calprotectin (CP) is released into the gut lumen during CDI to chelate zinc (Zn) and other essential nutrient metals. Yet, the impact of Zn limitation on C. difficile colonization is unknown. To define C. difficile responses to Zn limitation, we performed RNA sequencing on C. difficile exposed to CP. In medium containing CP, C. difficile upregulated genes involved in metal homeostasis and amino acid metabolism. To identify CP-responsive genes important during infection, we measured the abundance of select C. difficile transcripts in a mouse CDI model relative to expression in vitro Gene transcripts involved in selenium (Se)-dependent proline fermentation increased during infection and in response to CP. Increased proline fermentation gene transcription was dependent on CP Zn binding and proline availability, yet proline fermentation was only enhanced when Se was supplemented. CP-deficient mice could not restrain C. difficile proline fermentation-dependent growth, suggesting that CP-mediated Zn sequestration along with limited Se restricts C. difficile proline fermentation. Overall, these results highlight how C. difficile colonization depends on the availability of multiple nutrients whose abundances are dynamically influenced by the host response.IMPORTANCEClostridioides difficile infection (CDI) is the leading cause of postantibiotic nosocomial infection. Antibiotic therapy can be successful, yet up to one-third of individuals suffer from recurrent infections. Understanding the mechanisms controlling C. difficile colonization is paramount in designing novel treatments for primary and recurrent CDI. Here, we found that limiting nutrients control C. difficile metabolism during CDI and influence overall pathogen fitness. Specifically, the immune protein CP limits Zn availability and increases transcription of C. difficile genes necessary for proline fermentation. Paradoxically, this leads to reduced C. difficile proline fermentation. This reduced fermentation is due to limited availability of another nutrient required for proline fermentation, Se. Therefore, CP-mediated Zn limitation combined with low Se levels overall reduce C. difficile fitness in the intestines. These results emphasize the complexities of how nutrient availability influences C. difficile colonization and provide insight into critical metabolic processes that drive the pathogen's growth.

Sequestration of Zn into mixed pyrite-zinc sulfide framboids: A key to Zn cycling in the ocean?

Zinc (Zn) is an important micronutrient in the ocean, and fixation of Zn into organic, trace element-rich sediments is an important contributor to Zn cycling in the ocean. Framboidal sulfides are considered to be the major host for Zn in such settings. The sequestration of Zn into framboids via biotic or abiotic processes is not fully understood, which presents difficulties for interpretation of Zn isotope values in sediments. In this work, we describe a novel type of framboid with mixed pyrite and zinc sulfide (sphalerite or wurtzite) microcrystals from meta-pelites of the Otago Schist, New Zealand. A combination of optical microscopy, scanning electron microscopy (SEM) and nanoscale secondary ion mass spectrometry (NanoSIMS) were utilized to assess the association between Zn, pyrite and organic matter in framboids. The distribution of Zn in framboids is variable. Most pyrite microcrystals include minor amounts of Zn. Trace Zn is also observed to co-locate with organic matter, which occurs on the boundaries of pyrite microcrystals. Finally, Zn is found as single zinc sulfide microcrystals or zinc sulfide rims around pyrite microcrystals within individual framboids. These textures have not been recorded before, to our knowledge. The sequence of events that sequesters Zn into framboids may affect Zn isotope fractionation from seawater to continental margin sediments.