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.     

Phytoremediation of Heavy Metals: Physiological and Molecular Mechanisms

Heavy metals (HM) are a unique class of toxicants since they cannot be broken down to non-toxic forms. Concentration of these heavy metals has increased drastically, posing problems to health and environment, since the onset of the industrial revolution. Once the heavy metals contaminate the ecosystem, they remain a potential threat for many years. Some technologies have long been in use to remove, destroy and sequester these hazardous elements. Even though effective techniques for cleaning the contaminated soils and waters are usually expensive, labour intensive, and often disturbing. Phytoremediation, a fast-emerging new technology for removal of toxic heavy metals, is cost-effective, non-intrusive and aesthetically pleasing. It exploits the ability of selected plants to remediate pollutants from contaminated sites. Plants have inter-linked physiological and molecular mechanisms of tolerance to heavy metals. High tolerance to HM toxicity is based on a reduced metal uptake or increased internal sequestration, which is manifested by interaction between a genotype and its environment. The growing interest in molecular genetics has increased our understanding of mechanisms of HM tolerance in plants and many transgenic plants have displayed increased HM tolerance. Improvement of plants by genetic engineering, i.e., by modifying characteristics like metal uptake, transport and accumulation and plant’s tolerance to metals, opens up new possibilities of phytoremediation. This paper presents an overview of the molecular and physiological mechanisms involved in the phytoremediation process, and discusses strategies for engineering plants genetically for this purpose.     

Molecular mechanisms of heavy metal hyperaccumulation and phytoremediation

A relatively small group of hyperaccumulator plants is capable of sequestering heavy metals in their shoot tissues at high concentrations. In recent years, major scientific progress has been made in understanding the physiological mechanisms of metal uptake and transport in these plants. However, relatively little is known about the molecular bases of hyperaccumulation. In this paper, current progresses on understanding cellular/molecular mechanisms of metal tolerance/hyperaccumulation by plants are reviewed. The major processes involved in hyperaccumulation of trace metals from the soil to the shoots by hyperaccumulators include: (a) bioactivation of metals in the rhizosphere through root–microbe interaction; (b) enhanced uptake by metal transporters in the plasma membranes; (c) detoxification of metals by distributing to the apoplasts like binding to cell walls and chelation of metals in the cytoplasm with various ligands, such as phytochelatins, metallothioneins, metal-binding proteins; (d) sequestration of metals into the vacuole by tonoplast-located transporters. The growing application of molecular-genetic technologies led to the well understanding of mechanisms of heavy metal tolerance/accumulation in plants, and subsequently many transgenic plants with increased resistance and uptake of heavy metals were developed for the purpose of phytoremediation. Once the rate-limiting steps for uptake, translocation, and detoxification of metals in hyperaccumulating plants are identified, more informed construction of transgenic plants would result in improved applicability of the phytoremediation technology.     

丛枝菌根在植物修复重金属污染土壤中的作用

丛枝菌根（Arbuscular mycorrhizae，AM）是自然界中分布最广的一类菌根，AM真菌能与陆地上绝大多数的高等植物共生，常见于包括重金属污染土壤在内的各种生境中。在重金属污染条件下，AM真菌可以减轻重金属对植物的毒害，影响植物对重金属的吸收和转运，在重金属污染土壤的植物修复中显示出极大的应用潜力。重点介绍了AM真菌对植物重金属耐性的影响及其在植物提取和植物稳定中的应用等方面的进展，讨论了未来研究所面临的任务和挑战。     

植物根际促生菌及丛枝菌根真菌协助植物修复重金属污染土壤的机制

植物修复是一种前景广阔的重金属污染土壤的主要修复技术,在微生物的协助下效果更为显著。植物根际促生菌可通过分泌吲哚-3-乙酸(IAA)、产铁载体、固氮溶磷等方式促进植物生长、改善植物重金属耐受性,从而有效提高重金属污染土壤的植物修复效率。菌根真菌是土壤-植物系统中重要的功能菌群之一,可侵染植物根系改变根系形态和矿质营养状况,通过菌丝体吸附重金属,也可产生球囊霉素、有机酸、植物生长素等次生代谢产物改变重金属生物有效性。植物根际促生菌与丛枝菌根真菌可对植物产生协同促生作用,在重金属污染土壤修复中具有一定应用潜力。目前,国内外关于植物根际促生菌和丛枝菌根真菌互作已有大量研究,而二者的相互作用机理仍处于探索阶段。本文综述了近年来国内外植物根际促生菌和丛枝菌根真菌在重金属污染土壤植物修复中的作用机制,并对其研究前景进行展望。     

重金属污染的植物修复及相关分子机制

随着近代工业的发展,土壤重金属污染问题日益严重。重金属即使在极低浓度下仍然可以对人畜造成健康上的威胁,因此迫切需要有效的修复方法对土壤进行修复。生物修复,特别是植物修复目前已经成为重金属污染修复的重要手段之一,了解相关植物的重金属解毒和积累分子机制是提高修复效率、解决重金属污染问题的基础。文中以土壤修复方式为起点,结合植物吸收积累重金属以及解毒的相关分子机制研究,探讨了植物修复的发展现状以及趋势。     

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。     

Influence of Soil Chemistry and Plant Physiology in the Phytoremediation of Cu,Mn, and Zn

Different anthropogenic sources of metals can result from agricultural, industrial, military, mining and urban activities that contribute to environmental pollution. Plants can be grown for phytoremediation to remove or stabilize contaminants in water and soil. Copper (Cu), manganese (Mn) and zinc (Zn) are trace essential metals for plants, although their role in homeostasis in plants must be strictly regulated to avoid toxicity. In this review, we summarize the processes involved in the bioavailability, uptake, transport and storage of Cu, Mn and Zn in plants. The efficiency of phytoremediation depends on several factors including metal bioavailability and plant uptake, translocation and tolerance mechanisms. Soil parameters, such as clay fraction, organic matter content, oxidation state, pH, redox potential, aeration, and the presence of specific organisms, play fundamental roles in the uptake of trace essential metals. Key processes in the metal homeostasis network in plants have been identified. Membrane transporters involved in the acquisition, transport and storage of trace essential metals are reviewed. Recent advances in understanding the biochemical and molecular mechanisms of Cu, Mn and Zn hyperaccumulation are described. The use of plant-bacteria associations, plant-fungi associations and genetic engineering has opened a new range of opportunities to improve the efficiency of phytoremediation. The main directions for future research are proposed from the investigation of published results.     

我国土壤重金属污染植物吸取修复研究进展

我国从上世纪90年代中后期开始土壤重金属（含类金属砷）污染的植物吸取修复研究及技术探索,先后发现了一批具有较高研究价值和应用前景的铜、砷、镉、锰等重金属的积累或超积累植物,并从重金属耐性和超积累生理机制、植物吸取修复的根际过程与机制、吸取修复强化措施和修复植物处置与资源化利用等方面进行了研究,同时开展了植物吸取修复技术的示范与应用,已有一些较成功的植物修复工程应用案例,使我国重金属污染土壤植物修复技术,尤其是植物吸取修复技术在国际上产生了较强的影响力。本文就近年来我国土壤重金属污染植物吸取修复研究进展进行了综述,并对今后的发展趋势进行了展望。     

A Review on Mechanisms of Plant Tolerance and Role of Transgenic Plants in Environmental Clean-up

This article deals with the advances and implication of phytoremediation technologies with emphasis on remediation of toxic heavy metals from contaminated soil. Most of the conventional remedial technologies are expensive and inhibit the soil fertility and cause negative impacts on various ecosystem services. However, phytoremediation is a cost effective and ecofriendly approach, which does not adversely affect soil properties and ecosystem services. In recent years, major progresses have been made in understanding the physiological mechanisms of metal uptake and transport in hyper accumulators. However, the molecular mechanisms of metal uptake, translocation, accumulation and detoxification in plants and their further implication in transgenic development for efficient phytoremediation are not well understood. In view of above, present review article brings together existing bits of information to create a new direction for future research, critical gap in knowledge and a new perspective on remediation of toxic pollutants.     

Biochemical changes and adaptive strategies of plants under heavy metal stress

Heavy metal contamination of soil, aqueous waste stream and ground water causes major environmental and human health problems. Heavy metals are major environmental pollutants when they are present in high concentration in soil and show potential toxic effects on growth and development in plants. Due to unabated, indiscriminate and uncontrolled discharge of hazardous chemicals including heavy metals into the environment, plant continuously have to face various environmental constraints. In plants, seed germination is the first exchange interface with the surrounding medium and has been considered as highly sensitive to environmental changes. One of the crucial events during seed germination entails mobilization of seed reserves which is indispensable for the growth of embryonic axis. But, metabolic alterations by heavy metal exposure are known to depress the mobilization and utilization of reserve food by affecting the activity of hydrolytic enzymes. Some plants possess a range of potential mechanisms that may be involved in the detoxification of heavy metals by which they manage to survive under metal stress. High tolerance to heavy metal toxicity could rely either on reduced uptake or increase planned internal sequestration which is manifested by an interaction between a genotype and its environment. Such mechanism involves the binding of heavy metals to cell wall, immobilization, exclusion of the plasma membrane, efflux of these toxic metal ions, reduction of heavy metal transport, compartmentalization and metal chelation by tonoplast located transporters and expression of more general stress response mechanisms such as stress proteins. It is important to understand the toxicity response of plant to heavy metals so that we can utilize appropriate plant species in the rehabilitation of contaminated areas. Therefore, in the present review attempts have been made to evaluate the effects of increasing level of heavy metal in soils on the key behavior of hydrolytic and nitrogen assimilation enzymes. Additionally, it also provides a broad overview of the strategies adopted by plants against heavy metal stress.     

根系分泌物在重金属污染土壤生态修复中的作用机制研究进展

我国土壤重金属污染问题日益突出.作为一种绿色、安全的生物修复技术,植物修复技术备受关注.根系分泌物作为植物-土壤-微生物三者物质交换与信息传递的重要载体,是植物响应外界胁迫的重要生理生态指征,在植物修复过程中发挥关键作用.研究表明,根系分泌物能够有效调控根际微环境,提升植物抗逆能力,影响重金属在根际微域中的环境行为.传...     

Utilization of Multi-Tasking Non-Edible Plants for Phytoremediation and Bioenergy Source-A Review

Heavy metal contamination of land and freshwater resources is a serious concern worldwide. It adversely affects the health of animals, plants and humans. Therefore, remediation of toxic heavy metals must be highly considered. Unlike other techniques, phytoremediation is a holistic technology and can be used in large scale for soil remediation as it is costless, novel, environmentally-safe and solar-driven technology. Utilization of non-edible plants in phytoremediation is an ingenious technique as they are used to generate new bioenergy resources along with the remediation of contaminated soils. Some nonfood bioenergy crops such as Salix species, Miscanthus species, Populus species, Eucalyptus species, and Ricinus communis exhibit high capability to accumulate various metals and to grow in contaminated lands. However, there are still sustainable challenges facing coupling phytoremediation with bioenergy production from polluted lands. Therefore, there has long been a need for developing different strategies to resolve such challenges. In this article review, we will discuss the phytoremediation mechanism, the technique of phytoremediation coupling with bioenergy production, sustainable problems facing linking phytoremediation with energy production as well as possible strategies to enhance the efficiency of bioenergy plants for soil decontamination by improving their characteristics such as metal uptake, transport, accumulation, and tolerance.     

AM真菌对重金属污染土壤生物修复的应用与机理

土壤重金属污染威胁人类健康和整个生态系统,而高效、低耗、安全的生物修复技术显示出了极大的应用潜力,特别是利用植物-微生物共生体增强生物修复效应的应用.丛枝菌根(Arbuscular Mycorrhizae,AM)真菌是一类广泛分布于土壤生态系统中的有益微生物,能与90％以上的陆生高等植物形成共生体.研究发现,AM真菌能够增强宿主植物对土壤中重金属胁迫的耐受性.当前,利用AM真菌开展重金属污染土壤的生物修复已经引起环境学家和生态学家的广泛关注.基于此,围绕AM真菌在重金属污染土壤生物修复作用中的最新研究进展,从物理性防御体系的形成、对植物生理代谢的调控、生化拮抗物质的产生、基因表达的调控等角度探究AM真菌在重金属污染土壤生物修复中的作用机理,以期为利用AM真菌开展重金属污染的生物修复提供理论依据,并对本领域未来的发展和应用前景进行了展望.     

植物对重金属的吸收和分布

植物修复是利用植物来清除污染土壤中重金属的一项技术。该技术成功与否取决于植 物从土壤中吸取金属以及向地上部运输金属的能力。植物对金属的吸收主要取决于自由态离子活度。许多螯合剂能诱导植物对重金属的吸收。金属离子在液泡中的区域化分布是植物耐 重金属的主要原因。同时,细胞内的金属硫蛋白、植物螯合肽等蛋白质以及有机酸、氨基酸等在金属贮存和解毒方面也起重要作用。本文还论述了重金属在植物体内运输的生理及分子 方面的研究进展。     

植物对重金属的吸收和分布

植物修复是利用植物来清除污染土壤中重金属的一项技术。该技术成功与否取决于植物从土壤中吸取金属以及向地上部运输金属的能力。植物对金属的吸收主要取决于自由态离子活度。许多螯合剂能诱导植物对重金属的吸收。金属离子在液泡中的区域化分布是植物耐重金属的主要原因。同时,细胞内的金属硫蛋白、植物螯合脓等蛋白质以及有机酸、氨基酸等在金属贮存和解毒方面也起重要作用。本文还论述了重金属在植物体内运输的生理及分子方面的研究进展。     

Phytoremediation of Metals Using Transgenic Plants

An ideal plant for environmental cleanup can be envisioned as one with high biomass production, combined with superior capacity for pollutant tolerance, accumulation, and/or degradation, depending on the type of pollutant and the phytoremediation technology of choice. With the use of genetic engineering, it is feasible to manipulate a plant's capacity to tolerate, accumulate, and/or metabolize pollutants, and thus to create the ideal plant for environmental cleanup. In this review, we focus on the design and creation of transgenic plants for phytoremediation of metals. Plant properties important for metal phytoremediation are metal tolerance and accumulation, which are determined by metal uptake, root-shoot translocation, intracellular sequestration, chemical modification, and general stress resistance. If we know which molecular mechanisms are involved in these tolerance and accumulation processes, and which genes control these mechanisms, we can manipulate them to our advantage. This review aims to give a succinct overview of plant metal tolerance and accumulation mechanisms, and to identify possible strategies for genetic engineering of plants for metal phytoremediation. An overview is presented of what has been achieved so far regarding the manipulation of plant metal metabolism. In fact, both enhanced metal tolerance and accumulation have been achieved by overproducing metal chelating molecules (citrate, phytochelatins, metallothioneins, phytosiderophores, ferritin) or by the overexpression of metal transporter proteins. Mercury volatilization and tolerance was achieved by introduction of a bacterial pathway. The typical increase in metal accumulation as the result of these genetic engineering approaches is 2-to 3-fold more metal per plant, which could potentially enhance phytoremediation efficiency by the same factor. As for the applicability of these transgenics for environmental cleanup, results from lab and greenhouse studies look promising for several of these transgenics, but field studies will be the ultimate test to establish their phytoremediation potential, their competitiveness, and risks associated with their use.     

Opportunities and feasibilities for biotechnological improvement of Zn, Cd or Ni tolerance and accumulation in plants

Metals contaminate the soil when present in high concentrations causing soil and ultimately environmental pollution. “Phytoremediation” is the use of plants to remove pollutants from contaminated environments. Plants tightly regulate their internal metal concentrations in a process called “metal homeostasis”. Some species have evolved extreme tolerance and accumulation of Zn, Cd and Ni as a way to adapt to exposure to these metals. Such traits are beneficial for phytoremediation, however, most natural metal hyperaccumulator species are not adapted to agriculture and have low yields. A wealth of knowledge has been generated regarding metal homeostasis in plants, including hyperaccumulators, which can be used in phytoremediation of Zn, Cd and Ni. In this review, we describe the current state of Zn, Cd and Ni physiology in plants and the underlying molecular mechanisms. The ways to efficiently utilize this information in designing high biomass metal accumulator plants are discussed. The potential and application of genetic modification has extended our understanding about the mechanisms in plants dealing with the metal environment and has paved the way to achieve the goal of understanding metal physiology and to apply the knowledge for the containment and clean up of metal contaminated soils.     

植物螯合肽及其在重金属耐性中的作用

综述植物螯合肽的生物合成及其在重金属耐性中的作用．有毒重金属在土壤中的积累不仅影响作物的生长和产量形成,而且严重威胁农产品的安全性．植物对重金属的耐性和积累在种间和基因型之间存在着很大的差异,在重金属胁迫条件下植物螯合肽(PC)的合成是植物对胁迫的一种适应性反应,耐性基因型合成较多的PC谷胱苷肽是合成PC的前体,PC可与重金属螯合,并进一步转运至液泡贮存,使细胞质的重金属浓度降低,从而达到解毒效果．重金属诱导植物合成PC的遗传机理和生化途径有赖于分子生物学的深入研究,cD-敏感型拟南芥突变体Cad1-1(缺失GSH)和Cad2-1(缺失PC合成诱导酶)的分离及相关研究,佐证了PC在Cd-解毒中起关键作用．对PC在重金属污染土壤或水体的植物修复和农作物安全生产中的意义进行了讨论．     

芒属植物重金属耐性及其在矿山废弃地植被恢复中的应用潜力

芒属植物重金属耐性强,并且是重要的能源植物,其在矿山废弃地植被恢复中的应用备受关注.芒属植物对多种重金属耐性强,但不属于重金属超累积植物.目前的研究认为,根系代谢能力强、根际存在多种共生微生物及抗氧化和光合作用能力强是芒属植物重金属耐性强的重要原因,但更为全面的耐性机理需要深入研究.芒属植物在矿山废弃地植被恢复的应用潜力大,可以清除土壤重金属、改善土壤性质和促进生物多样性发展.本文总结分析了芒属植物生物学特性、重金属耐性特点、机理及其在矿山废弃地植被恢复中的应用潜力,提出了应用芒属植物进行矿山废弃地植被恢复的基本思路,并对芒属植物的重金属耐性机理及应用的未来研究方向进行展望,以期为利用芒属植物开展矿山废弃地植被恢复提供借鉴.