Research Advances on the Mechanisms of Heavy Metal Tolerance in Plants

Heavy metals impact on the cytoplasmic function in a number of different ways, principally by their binding to protein sulflhdryl groups, by producing a deficiency of essential ions and, eventually, by substituting the essemial ions. Other modes of toxicity are possible, including disruption of cell transport processes and oxidative damage by free radicals generated by metal redox cycling. Plants have developed a variety of biochemical defense strategies to prevent heavy metal poisoning. The possible defense mechanism in plant may involve: metal binding to cell walls, avoidance of uptake these toxic metal ions, reduction of heavy metal transport across the cell membrane, active efflux, compartmentalization and metal chelation. Phytochelatins that can tightly bind and sequester metals may play an important role in the accumulation of heavy metals and preventing them from entering the cell metabolic pathway, the rates of high molecular weight (HMW) metal phytochelatin complexes (Cd-Sa-complex) formation may be an important determinant of the plant tolerance. In addition, plants possess several antioxidant defense systems to protect themselves from the oxidative stress by heavy metals.     

植物耐重金属机理研究进展

由于工业“三废”和机动车尾气的排放、污水灌溉及农药、除草剂和化肥的使用,严重地污染了土壤、水质和大气,其中土壤中的重金属（Ｈｇ、Ｃｄ、Ａｓ、Ｃｕ和Ａｌ）污染更为严重［１］。重金属在植物根、茎、叶及籽粒中的大量累积,不仅严重地影响植物的生长和发育［１～...     

Bacterial resistances to mercury and copper.

Heavy metals are toxic to living organisms. Some have no known beneficial biological function, while others have essential roles in physiological reactions. Mechanisms which deal with heavy metal stress must protect against the deleterious effects of heavy metals, yet avoid depleting the cell of a heavy metal which is also an essential nutrient. We describe the mechanisms of resistance in Escherichia coli to two different heavy metals, mercury and copper. Resistance of E. coli to mercury is reasonably well understood and is known to occur by transport of mercuric ions into the cytoplasmic compartment of the bacterial cell and subsequent reductive detoxification of mercuric ions. Recent mutational analysis has started to uncover the mechanistic detail of the mercuric ion transport processes, and has shown the essential nature of cysteine residues in transport of Hg(II). Resistance to copper is much less well understood, but is known to involve the increased export of copper from the bacterial cell and modification of the copper; the details of the process are still being elucidated. Expression of both metal resistance determinants is regulated by the corresponding cation. In each case the response enables the maintenance of cellular homeostasis for the metal. The conclusions drawn allow us to make testable predictions about the regulation of expression of resistance to other heavy metals.     

Modification of heavy metal uptake efficiency by modified chitosan/anionic surfactant systems

The presence of toxic heavy metals in natural environments entails a potential health hazard for humans. Metal contaminants in these environments are usually tightly bound to colloidal particles and organic matter. On the other hand, the potential of these metals towards chelation by different chelating agents presents a good characteristic for their removal from the environment. On this basis, two chitosan/anionic surfactant complexes were prepared and evaluated for their ability to remove heavy metals from aqueous solutions. The experimental results of the uptake of metal ions including Cu²⁺, Sn²⁺, Co²⁺ and Ni²⁺ are reported in this study. The results show that modified chitosan with short‐spacer group cross‐linkers has a higher potential for heavy metal uptake than long‐chain cross‐linker‐modified chitosan. Also, increasing the electronegativity of the heavy metals increases their uptake from the medium. Increasing the time of exposure of the heavy metals to the modified polymer increases the efficiency of the metal uptake process.     

Resistance to cadmium, cobalt, zinc, and nickel in microbes.

The divalent cations of cobalt, zinc, and nickel are essential nutrients for bacteria, required as trace elements at nanomolar concentrations. However, at micro- or millimolar concentrations, Co2+, Zn2+, and Ni2+ (and "bad ions" without nutritional roles such as Cd2+) are toxic. These cations are transported into the cell by constitutively expressed divalent cation uptake systems of broad specificity, i.e., basically Mg2+ transport systems. Therefore, in case of a heavy metal stress, uptake of the toxic ions cannot be reduced by a simple down-regulation of the transport activity. As a response to the resulting metal toxicity, metal resistance determinants evolved which are mostly plasmid-encoded in bacteria. In contrast to that of the cation Hg2+, chemical reduction of Co2+, Zn2+, Ni2+, and Cd2+ by the cell is not possible or sensible. Therefore, other than mutations limiting the ion range of the uptake system, only two basic mechanisms of resistance to these ions are possible (and were developed by evolution): intracellular complexation of the toxic metal ion is mainly used in eucaryotes; the cadmium-binding components are phytochelatins in plant and yeast cells and metallothioneins in animals, plants, and yeasts. In contrast, reduced accumulation based on an active efflux of the cation is the primary mechanism developed in procaryotes and perhaps in Saccharomyces cerevisiae. All bacterial cation efflux systems characterized to date are plasmid-encoded and inducible but differ in energy-coupling and in the number and types of proteins involved in metal transport and in regulation. In the gram-positive multiple-metal-resistant bacterium Staphylococcus aureus, Cd2+ (and probably Zn2+) efflux is catalyzed by the membrane-bound CadA protein, a P-type ATPase. However, a second protein (CadC) is required for full resistance and a third one (CadR) is hypothesized for regulation of the resistance determinant. The czc determinant from the gram-negative multiple-metal-resistant bacterium Alcaligenes eutrophus encodes proteins required for Co2+, Zn2+, and Cd2+ efflux (CzcA, CzcB, and CzcC) and regulation of the czc determinant (CzcD). In the current working model CzcA works as a cation-proton antiporter, CzcB as a cation-binding subunit, and CzcC as a modifier protein required to change the substrate specificity of the system from Zn2+ only to Co2+, Zn2+, and Cd2+.     

Efflux-mediated heavy metal resistance in prokaryotes

What makes a heavy metal resistant bacterium heavy metal resistant? The mechanisms of action, physiological functions, and distribution of metal-exporting proteins are outlined, namely: CBA efflux pumps driven by proteins of the resistance-nodulation-cell division superfamily, P-type ATPases, cation diffusion facilitator and chromate proteins, NreB- and CnrT-like resistance factors. The complement of efflux systems of 63 sequenced prokaryotes was compared with that of the heavy metal resistant bacterium Ralstonia metallidurans. This comparison shows that heavy metal resistance is the result of multiple layers of resistance systems with overlapping substrate specificities, but unique functions. Some of these systems are widespread and serve in the basic defense of the cell against superfluous heavy metals, but some are highly specialized and occur only in a few bacteria. Possession of the latter systems makes a bacterium heavy metal resistant.     

From industrial sites to environmental applications with <Emphasis Type="Italic">Cupriavidus metallidurans</Emphasis>

Cupriavidus metallidurans CH34 and related strains are adapted to metal contaminated environments. A strong resistance to environmental stressors and adaptation make it ideal strains for survival in decreasing biodiversity conditions and for bioaugmentation purposes in environmental applications. The soil bacterium C. metallidurans is able to grow chemolithoautotrophically on hydrogen and carbon dioxide allowing a strong resilience under conditions lacking organic matter. The biofilm growth on soil particles allows coping with starvation or bad conditions of pH, temperature and pollutants. Its genomic capacity of two megaplasmids encoding several heavy metal resistance operons allowed growth in heavy metal contaminated habitats. In addition its specific siderophores seem to play a role in heavy metal sequestration besides their role in the management of bioavailable iron. Efflux ATPases and RND systems pump the metal cations to the membrane surface where polysaccharides serve as heavy metal binding and nucleation sites for crystallisation of metal carbonates. These polysaccharides contribute also to flotation under specific conditions in a soil-heavy metals–bacteria suspension mixture. An inoculated moving bed sand filter was constructed to treat heavy metal contaminated water and to remove the metals in the form of biomass mixed with metal carbonates. A membrane based contactor allowed to use the bacteria as well in a versatile wastewater treatment system and to grow homogeneously formed heavy metal carbonates. Its behaviour toward heavy metal binding and flotation was combined in a biometal sludge reactor to extract and separate heavy metals from metal contaminated soils. Finally its metal-induced heavy metal resistance allowed constructing whole cell heavy metal biosensors which, after contact with contaminated soil, waste, solids, minerals and ashes, were induced in function of the bioavailable concentration (Cd, Zn, Cu, Cr, Co, Ni, Tl, Pb and Hg) in the solids and allowed to investigate the speciation of immobilization of those metals. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Physiological responses of plants to heavy metals and the quantification of tolerance and toxicity

Abstract

Techniques available for assessing the tolerance of plants to heavy metal toxins are reviewed. All are based on physiological responses and range from long-term growth trials in metal-contaminated substrates, to rapid cytological tests. Problems associated with the ecophysiological interpretation of in vitro measurements of tolerance are considered. The implications of multiple tolerance, co-tolerance, constitutional tolerance, inducible tolerance and possible stimulatory effects of metals on plant responses are discussed.     

Biosorption of Cu(II) and Pb(II) ions from aqueous solution by natural spider silk

Aside from its excellent mechanical properties, spider silk (SS) would offer an active surface for heavy metal interaction due to its rich protein structure. The present study describes the potential use of natural (SS) as a sorbent of heavy metals from aqueous solutions. Single and multi-species biosorption experiments of heavy metals by natural SS were conducted using batch and column experiments. The biosorption kinetics, in general, was found to follow the second-order rate expression, and the experimental equilibrium biosorption data fitted reasonably well to Freundlich isotherm. From the Freundlich isotherm, the biosorption capacities of Cu(II) and Pb(II) ions onto SS were found as 0.20 and 0.007 mmol g?1, respectively. The results showed a decrease in the extent of metal ion uptake with lowering the pH.     

The relationship between co-exposure to multiple heavy metals and liver damage

BackgroundThe impact of heavy metal exposure on human health has attracted widespread attention of researchers, and the impact of heavy metal exposure on liver function has also been confirmed, however, more attention is paid to the impact of single or two heavy metal exposures, and most epidemiological studies focus on heavy metal pollution areas. In this study, rural residents in non-heavy metal-contaminated areas in Northwest China were selected as the research objects to explore the comprehensive effects of co-exposure to multiple heavy metals on the liver, which can provide certain reference and support for related research.ObjectivesThis study used a Bayesian nuclear machine model (BKMR) to evaluate the relationship between exposure to heavy metal mixtures and indicators of liver function in a population in rural Northwest China.ResultsExposure to higher concentrations of metal mixtures was positively correlated with total bilirubin, direct bilirubin, and aspartate aminotransferase, and negatively correlated with alanine aminotransferase, with Pb contributing the most to indicators of liver function. We also observed a possible interaction of Cd with other heavy metals in the effect of heavy metal mixtures on DB levels.ConclusionsConcurrent exposure to higher concentrations of heavy metal mixtures (Cr, Co, Cd, and Pb) in rural China was associated with indicators representing poor liver function, of which the effect of lead on liver function should be focused. More prospective epidemiological studies and animal experiments need to be carried out to determine this relationship and possible mechanism.     

细菌金属离子外排系统及金属稳态调控

铁、铜、锌、锰等金属离子是各类生物体生存和增殖所必需的微量元素,可影响生物体内蛋白酶活性、免疫反应、生理过程和抗感染机制。细菌感染过程中,宿主可通过限制或提高体内环境中金属离子的浓度来抑制细菌增殖,与此同时,细菌进化出各种转运系统以适应宿主体内金属离子水平的变化。由于不同细菌的金属离子外排系统在结构和生化特性上存在变异,它们呈现出独特的金属离子外排模式。本文根据现有文献报道及本团队研究结果,对铁、铜、锌和锰离子的细菌外排系统进行讨论和总结,旨在综述目前对细菌金属离子稳态调控机制研究进展的认识,为深入理解细菌金属稳态调控相关机制提供参考。     

Isolation of a novel<Emphasis Type="Italic"> Thermus thermophilus</Emphasis> metal efflux protein that improves<Emphasis Type="Italic"> Escherichia coli</Emphasis> growth under stress conditions

The mechanisms of metal ion transport in thermophilic organisms are poorly understood. Phage display-based screening of a Thermus thermophilus genomic library in Escherichia coli led to the identification of a novel metal cation efflux protein. The Thermus protein showed extensive sequence and putative structural conservation to Czr and Czc proteins in mesophilic bacterial and mammalian species. Expression of the gene in E. coli led to increased resistance to zinc and cadmium ions, but not to cobalt, in an effect that was apparently caused by increased efflux of metals from the cell. This increased resistance was inducible by zinc and cadmium and, to a lesser extent, by cobalt. Furthermore, E. coli cells containing the Thermus gene exhibited improved cell physiology and delayed cell lysis during recombinant protein production, leading to accumulation of higher levels of recombinant protein. The molecular basis and potential application of the findings are discussed.     

MRP proteins as potential mediators of heavy metal resistance in zebrafish cells

Acquired resistance of mammalian cells to heavy metals is closely relevant to enhanced expression of several multidrug resistance-associated proteins (MRP), but it remains unclear whether MRP proteins confer resistance to heavy metals in zebrafish. In this study, we obtained zebrafish (Danio rerio) fibroblast-like ZF4 cells with resistance to toxic heavy metals after chronic cadmium exposure and selection for 6months. These cadmium-resistant cells (ZF4-Cd) were maintained in 5μM cadmium and displayed cross-resistance to cadmium, mercury, arsenite and arsenate. ZF4-Cd cells remained the resistance to heavy metals after protracted culture in cadmium-free medium. In comparison with ZF4-WT cells, ZF4-Cd cells exhibited accelerated rate of cadmium excretion, enhanced activity of MRP-like transport, elevated expression of abcc2, abcc4 and mt2 genes, and increased content of cellular GSH. Inhibition of MRP-like transport activity, GSH biosynthesis and GST activity significantly attenuated the resistance of ZF4-Cd cells to heavy metals. The results indicate that some of MRP transporters are involved in the efflux of heavy metals conjugated with cellular GSH and thus play crucial roles in heavy metal detoxification of zebrafish cells.     

Theczc operon ofAlcaligenes eutrophus CH34: from resistance mechanism to the removal of heavy metals

Summary The plasmid-borneczc operon ensures for resistance to Cd²⁺, Zn²⁺ and Co²⁺ ions through a tricomponent export pathway and is associated to various conjugative plasmids ofA. eutrophus strains isolated from metal-contaminated industrial areas. Theczc region of pMOL30 was reassessed especially for the segments located upstream and downstream the structural genesczc CBA. In cultures grown with high concentrations of heavy metals,czc-mediated efflux of cations is followed by a process of metal bioprecipitation. These observations led to the development of bioreactors designed for the removal of heavy metals from polluted effluents.     

植物对重金属耐性的分子生态机理

植物适应重金属元素胁迫的机制包括阻止和控制重金属的吸收、体内螯合解毒、体内区室化分隔以及代谢平衡等。近年来,随着分子生物学技术在生态学研究中的深入应用,控制这些过程的分子生态机理逐渐被揭示出来。菌根、根系分泌物以及细胞膜是控制重金属进入植物根系细胞的主要生理单元。外生菌根能显著提高寄主植物的重金属耐性,根系分泌物通过改变根际pH、改变金属物质的氧化还原状态和形成络合物等机理减少植物对重金属的吸收。目前,控制菌根和根系分泌物重金属抗性的分子生态机理还不清楚。但细胞膜跨膜转运器已得到深入研究,相关金属离子转运器被鉴定和分离,一些控制基因如铁锌控制运转相关蛋白(ZIP)类、自然抵抗相关巨噬细胞蛋白(Nramp)类、P_1B-type ATPase类基因已被发现和克隆。金属硫蛋白(MTs)、植物螯合素(PCs)、有机酸及氨基酸等是植物体内主要的螯合物质,它们通过螯合作用固定金属离子,降低其生物毒性或改变其移动性。与MTs合成相关的MT-like基因已经被克隆,PCs合成必需的植物螯合素合酶(PCS), 即γ-Glu-Cys二肽转肽酶(γ-ECS) 的编码基因已经被克隆,控制麦根酸合成的氨基酸尼克烟酰胺(NA)在重金属耐性中的作用和分子机理也被揭示出来。ATP 结合转运器(ABC)和阳离子扩散促进器(CDF) 是植物体内两种主要膜转运器,通过它们和其它跨膜方式,重金属被分隔贮藏于液泡内。控制这些蛋白转运器合成的基因也已经被克隆,在植物中的表达证实其与重金属的体内运输和平衡有关。热休克蛋白(HSP)等蛋白类物质的产生是一种重要的体内平衡机制,其分子机理有待进一步研究。重金属耐性植物在这些环节产生了相关响应基因或功能蛋白质,分子克隆和转基因技术又使它们在污染治理上得到了初步的应用。     

植物-菌根真菌联合修复重金属污染土壤

菌根是菌根真菌侵染植物根系后在植物根部形成的共生结构。菌根技术作为一种生物强化技术应用于重金属污染土壤的植物修复已引起研究者的广泛关注。目前大量研究表明菌根能强化植物对重金属的转运、富集及根系稳定化过程,并通过促进营养物质的吸收利用、稳定细胞内氧化还原平衡、调控抗逆性相关基因的表达以及改善根际微生态环境等方式提升寄主植物的抗逆性。本文在介绍菌根真菌在植物修复重金属污染的联合过程中的作用效应及机制的基础上,分析了目前限制该技术应用的瓶颈问题以及未来的研究方向,为植物-菌根真菌联合修复的推广应用提供理论基础。     

Metal resistance in Candida biofilms

Yeasts are often successful in metal-polluted environments; therefore, the ability of biofilm and planktonic cell Candida tropicalis to endure metal toxicity was investigated. Fifteen water-soluble metal ions, chosen to represent groups 6A to 6B of the periodic table, were tested against this organism. With in vitro exposures as long as 24 h, biofilms were up to 65 times more tolerant to killing by metals than corresponding planktonic cultures. Of the most toxic heavy metals tested, only very high concentrations of Hg2+, CrO4 (2-) or Cu2+ killed surface-adherent Candida. Metal-chelator precipitates could be formed in biofilms following exposure to the heavy metals Cu2+ and Ni2+. This suggests that Candida biofilms may adsorb metal cations from their surroundings and that sequestration in the extracellular matrix may contribute to resistance. We concluded that biofilm formation may be a strategy for metal resistance and/or tolerance in yeasts.     

Molecular Mechanisms and Genetic Basis of Heavy Metal Tolerance/Hyperaccumulation in Plants

Phytoremediation has gained increased attention as a cost-effective method for the remediation of heavy metal-contaminated sites. Because some plants possess a range of potential mechanisms that may be involved in the detoxification of heavy metals, they manage to survive under metal stresses. High tolerance to heavy metal toxicity could rely either on reduced uptake or increased plant internal sequestration,which is manifested by an interaction between a genotype and its environment. The growing application of molecular genetic technologies has led to increased understanding of mechanisms of heavy metal tolerance/accumulation in plants and, subsequently, many transgenic plants with increased heavy metal resistance,as well as increased uptake of heavy metals, have been developed for the purpose of phytoremediation. In the present review, our major objective is to concisely evaluate the progress made so far in understanding the molecular/cellular mechanisms and genetic basis that control the uptake and detoxification of metals by plants.     

Advances in Cotton Tolerance to Heavy Metal Stress and Applications to Remediate Heavy Metal-Contaminated Farmland Soil

Heavy metal-contaminated soil is one of the major environmental pollution problems of agricultural production and human health in the world. Remediation of heavy metals in soil is one of the most popular research subjects. Different remediation strategies have been reported to remove heavy metals from contaminated soil, among which phytoremediation is the most important one. Compared with other major crops, cotton shows the strongest and most widespread resistance to abiotic stresses, such as heavy metals. Although heavy metal stress adversely affects the growth and development of cotton, cotton possesses a set of sophisticated stress-resistance strategies. As the main product of cotton is nonedible fibers, which have a large biomass and strong heavy metal absorption and enrichment capacities, cotton is an ideal crop to restore heavy metal-contaminated soils and has unique advantages in terms of both ecological and economic benefits, with great application prospects. In this review, based on domestic and foreign research results in recent years, the effects of heavy metals on cotton growth and product quality were analyzed, the heavy metal absorption, accumulation, translocation and enrichment characteristics of cotton plants were summarized, and the adaptation and tolerance mechanisms of cotton to heavy metals were explored. Furthermore, the view that cotton is an effective crop to remediate heavy metal pollution in farmland soil has been proposed, and popularization and application suggestions for planting cotton to repair heavy metal pollution have been put forward to provide a reference for the comprehensive evaluation of the economic feasibility of cotton to repair heavy metal pollution in farmland soil.