植物螯合肽及其功能

植物螯合肽（phytochelatin,PC）是一类富含Cys、由PC合酶以GSH为底物催化合成的小分子多肽,能通过Cys的-SH络合重金属.研究PC的合成机理及其重金属解毒机制、研究PC合酶和PC合酶基因的表达模式及其功能对于运用植物修复技术治理重金属污染的土壤和水体具有重要意义.     

植物络合素和植物络合素合酶的研究

植物络合素（Phytochelatins,PCs)是由于重金属离子诱导而在植物体内合成的一类小分子多肽,其结构式为（γ-Glu-Cys)n-Gly,(n=2-11);PCs能够螯合重金属,从而起到对对重金属解毒的作用,PCs并非基因的直接产物,而是由植物络合素合酶（phytochelatin syn-thase,PCS),以GSH为底物催化合成的;植物络合素合酶基因的表达是组成型的,重金属离子能够活化PCS,诱导PCs的合成。1989年,人们首次报道得到了部分纯化的PCS,10年后,3个研究小组分别于1999年同时克隆和鉴定了编码PCS的基因,这些结果不仅对于研究PCs的合成途径和模型的建立及植物抗重金属机制的探讨有重要意义,而且在利用基因工程改良植物抗重金属能力和净化环境污染方面有应用前景。     

植物防御素调控水稻镉积累的新机制

镉是我国农产品的主要重金属污染物之一。随着我国土壤重金属污染问题日益突出, 包括稻米在内的农产品重金属超标时常发生。如何防控重金属在作物可食部位的积累, 在保证农产品安全的同时将农田重金属进行移除修复, 已成为我国农业生产急需解决的问题。最近, 中科院上海生命科学院植物生理生态所龚继明研究组和中国水稻所钱前研究组克隆到1个特异调控镉在水稻(Oryza sativa)叶片中积累的主效QTL基因CAL1。CAL1编码1个植物防御素类似蛋白, 通过与镉进行螯合, 将镉从维管束木质部薄壁细胞中分泌出来, 进入木质部参与长距离转运, 从而定向调控镉在水稻叶片等营养器官的积累而不影响籽粒镉的积累。该研究加深了人们对重金属镉在植物体内的转运和再分配机理的认识, 同时也为培育秸秆镉高积累而籽粒镉含量达标的“修复型”水稻品种提供有价值的新基因。研究成果具有重要的理论意义和应用价值。     

构树转录因子BpbZIP1的鉴定及镉胁迫响应分析

重金属污染不仅影响土壤有效面积,限制植被分布,也会对食物链和人体健康造成危害,其中镉（Cd）污染尤为突出。选择重金属耐受性强的植物应用于尾矿区的土壤修复亟待进行。构树（Broussonetia papyrifera）是重金属污染土壤的先锋树种,为探明构树响应重金属胁迫的分子机制,本研究从构树中克隆获得1个碱性亮氨酸拉链（basic leucine zipper,bZIP）转录因子（命名为BpbZIP1）,对其基本生物信息、Cd胁迫响应及转化酵母的功能进行分析,预测BpbZIP1响应Cd的功能。结果显示,BpbZIP1基因开放阅读框长1 713 bp,编码的蛋白含570个氨基酸,分子量为62 902.38 Da,等电点为4.62。与拟南芥（Arabidopsis thaliana）AtbZIP1具有较近的进化关系。在150 μmol·L^-1 CdCl₂处理下,BpbZIP1基因能被不同程度的诱导表达,在3 h时BpbZIP1在根中的表达为对照的17.4倍。将BpbZIP1转入酵母能显著提高转基因酵母的抗Cd能力,当浓度高于0.6%时,转基因酵母的生长活力是对照的1.54~1.71倍。以上结果表明BpbZIP1基因能积极响应Cd胁迫,其表达可改善Cd胁迫耐受力,是构树Cd胁迫响应的重要基因。     

植物重金属超富集机理研究进展

植物超富集重金属机理主要涉及植物对金属离子高的吸收、运输能力,区域化作用及螯合作用等方面,其中跨膜运载蛋白的表达、调控对重金属超富集这一特性起了关键作用。金属阳离子运载蛋白家族主要包括CDF家族、NRAMP家族和ZIP家族等,在超富集植物中已克隆出多个家族的金属运载蛋白基因,这些基因的过量表达对重金属在细胞中的运输、分布和富集及提高植物的抗性方面发挥了重要作用。综述了近年来研究重金属超富集植物吸收、转运和贮存Zn、Ni、Cd等重金属的生理和分子机制所取得的主要进展。     

梯棱羊肚菌镉胁迫响应基因ATX1功能分析

梯棱羊肚菌Morchella importuna是一种可以大田覆土栽培的珍稀食用菌,而土壤重金属污染状况日益严重,对梯棱羊肚菌菌丝生长和子实体产品质量安全构成了潜在的威胁。本研究先采用镉离子胁迫处理梯棱羊肚菌菌丝体,RT-PCR检测发现候选基因ATX1的表达量显著下调。克隆梯棱羊肚菌ATX1基因,对ATX1p蛋白结构进行功能预测,发现ATX1p可能与铜离子转运及重金属胁迫相关。然后分别构建ATX1的超表达和RNAi基因沉默载体,采用农杆菌介导的转化方法,将其转入梯棱羊肚菌同核体菌株A50中,分别筛选到4个ATX1表达显著上调的超表达转化子和4个ATX1表达显著下调的RNAi基因沉默转化子,镉敏感性检测发现ATX1的RNAi基因沉默转化子表现为镉抗性增强,而ATX1超表达转化子则表现为镉抗性减弱。结果表明,梯棱羊肚菌ATX1基因表达与镉抗性呈负相关,ATX1p可能在梯棱羊肚菌镉胁迫响应过程中发挥着某种重要作用。     

镉污染水稻种子内生细菌的分离及其耐镉性和植物促生性研究

【目的】植物内生细菌在促进植物生长和重金属污染土壤修复中发挥着重要作用。本研究对镉污染水稻种子内生细菌进行分离和功能鉴定,从而确定其镉耐受性、植物促生性以及对镉胁迫下水稻种子萌发和幼苗生长的影响。【方法】采用乙醇-次氯酸钠联合灭菌法对水稻种子进行表面灭菌,采用标准平板培养法对内生细菌进行分离、纯化;对菌株16SrRNA基因进行扩增和测序,鉴定菌株的分类学地位;运用微量稀释法、电感耦合等离子体质谱法、比色法测定内生细菌的镉耐受性、镉去除率和植物促生性;采用浸种法进行菌剂侵染,观察施加菌剂对水稻种子萌发、幼苗生长和镉积累的影响。【结果】从两个水稻品种的种子中共分离得到133株内生细菌,分属于12个属的24个种。在低镉品种728B中分离得到83个菌株,主要包括假单胞菌属（34.94%）、芽孢杆菌属（28.92%）和类芽孢杆菌属（10.84%）菌株。在高镉品种博B中分离得到50个菌株,主要包括泛菌属（40%）、短小杆菌属（22%）和微杆菌属（12%）菌株。对24个代表性菌株的镉耐受性进行测定,发现镉离子对菌株的最小抑制浓度介于80–2 560μmol/L,其中芽孢杆菌HNR-4具有最高的镉耐受...     

强化龙葵富集镉根际促生菌的分离、筛选与鉴定

【背景】植物-微生物联合修复土壤重金属污染日渐兴起,获取与超富集植物高效互作的微生物是实现联合修复技术进步的关键。龙葵在镉农田污染修复中广泛应用。【目的】筛选可促进龙葵生长及富集镉的耐镉根际促生菌。【方法】从龙葵根际土分离耐镉菌株,筛选具有良好促生特性的菌株,水培试验考察镉胁迫下菌株对植物生长及镉富集能力的影响,确定可促进龙葵生长及富集镉的微生物菌株,通过生理生化特性和16S rRNA基因序列分析进行菌株的初步鉴定。【结果】分离得到NT1、AXY1、AW2和AW1四株强化龙葵富集镉促生菌,经鉴定分别为Lysinibacillus sp.、Beijerinckia fluminensis、Achromobacter animicus和Herbaspirillum huttiense。上述菌株均可有效促进龙葵生长,增加其株高和干物质积累,提高地上部镉富集量。其中,NT1可使株高、地上部干重分别增加31.33%和62.65%,AW2可使地上部镉富集量增加37.29%。【结论】筛选所得菌株可为提高植物修复效率提供实践依据,为研制田间施用生态功能菌剂做铺垫,用于农田镉污染的微生物-龙葵联合原位修复。     

超富集植物遏蓝菜对重金属吸收、运输和累积的机制

遏蓝菜Thlaspi caerulescens可以在其地上部累积大量重金属如锌、镉等,是公认的超富集植物。由于该植物生物量小,不宜直接用于重金属污染的土壤植物修复,而被广泛作为一种模式植物来进行重金属富集机制研究。遏蓝菜对重金属离子的累积大致经过螯合剂解毒、地上部长距离运输以及在液泡中的储存等生理过程。已经发现的植物体内的金属螯合剂——有机酸、氨基酸、植物络合素(PCs)、金属硫蛋白(MT)和尼克烟酰胺NA等,区室化以及长距离运输相关的转运蛋白——ZIP(ZRT/IRTlike protein)、CDF(Cation diffusion facilitator)、Nramp(Natural resistance and macrophage protein)和HMA(Heavy metal ATPase)等家族,以上各种基因、多肽与蛋白等共同参与了植物对金属累积与耐受过程并发挥各自重要的作用。以下主要介绍了遏蓝菜重金属超富集相关的基因、多肽和蛋白,以及它们在重金属螯合作用和运输过程中的功能。     

湖南柿竹园矿区土壤重金属含量及植物吸收特征

矿区重金属污染十分严重,寻找和发现适合当地气候与土壤条件的重金属耐性植物是矿区植被恢复和污染土壤修复的前提。对我国湖南柿竹园有色金属矿区调查发现,该地区选矿厂的重金属污染问题普遍比尾砂库严重。选矿厂土壤砷、镉、铅、锌严重超标,尾砂库周围也受到不同程度的重金属污染。土壤重金属胁迫效应影响着植物物种分布,选矿厂物种分布较少,相比之下尾砂库的植物多样性较为丰富。柿竹园矿区植物对重金属的吸收表现为富集型(如蜈蚣草Pteris Vittata L .和苎麻Boehmerianivea (L .) Gaud.)、根部囤积型(如攀倒甑Patrinia villosa和木贼Equisetum hyemale)和规避型(如蔓出卷柏Selaginelladavidii Franch和芒草Miscanthus sinensis Andlerss)等3种类型。     

Understanding molecular mechanisms for improving phytoremediation of heavy metal-contaminated soils

Heavy metal pollution of soil is a significant environmental problem with a negative potential impact on human health and agriculture. Rhizosphere, as an important interface of soil and plants, plays a significant role in phytoremediation of contaminated soil by heavy metals, in which, microbial populations are known to affect heavy metal mobility and availability to the plant through release of chelating agents, acidification, phosphate solubilization and redox changes, and therefore, have potential to enhance phytoremediation processes. Phytoremediation strategies with appropriate heavy metal-adapted rhizobacteria or mycorrhizas have received more and more attention. In addition, some plants possess a range of potential mechanisms that may be involved in the detoxification of heavy metals, and 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.A coordinated network of molecular processes provides plants with multiple metal-detoxifying mechanisms and repair capabilities. The growing application of molecular genetic technologies has led to an 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. This article reviews advantages, possible mechanisms, current status and future direction of phytoremediation for heavy-metal–contaminated soils.     

A novel heavy metal ATPase peptide from <Emphasis Type="Italic">Prosopis juliflora</Emphasis> is involved in metal uptake in yeast and tobacco

Heavy metal pollution of agricultural soils is one of the most severe ecological problems in the world. Prosopis juliflora, a phreatophytic tree species, grows well in heavy metal laden industrial sites and is known to accumulate heavy metals. Heavy Metal ATPases (HMAs) are ATP driven heavy metal pumps that translocate heavy metals across biological membranes thus helping the plant in heavy metal tolerance and phytoremediation. In the present study we have isolated and characterized a novel 28.9 kDa heavy metal ATPase peptide (PjHMT) from P. juliflora which shows high similarity to the C-terminal region of P_1B ATPase HMA1. It also shows the absence of the invariant signature sequence DKTGT, and the metal binding CPX motif but the presence of conserved regions like MVGEGINDAPAL (ATP binding consensus sequence), HEGGTLLVCLNS (metal binding domain) and MLTGD, GEGIND and HEGG motifs which play important roles in metal transport or ATP binding. PjHMT, was found to be upregulated under cadmium and zinc stress. Heterologous expression of PjHMT in yeast showed a higher accumulation and tolerance of heavy metals in yeast. Further, transgenic tobacco plants constitutively expressing PjHMT also showed increased accumulation and tolerance to cadmium. Thus, this study suggests that the transport peptide from P. juliflora may have an important role in Cd uptake and thus in phytoremediation.     

Identification of Thlaspi caerulescens genes that may be involved in heavy metal hyperaccumulation and tolerance. Characterization of a novel heavy metal transporting ATPase

Thlaspi caerulescens is a heavy metal hyperaccumulator plant species that is able to accumulate extremely high levels of zinc (Zn) and cadmium (Cd) in its shoots (30,000 microg g(-1) Zn and 10,000 microg g(-1) Cd), and has been the subject of intense research as a model plant to gain a better understanding of the mechanisms of heavy metal hyperaccumulation and tolerance and as a source of genes for developing plant species better suited for the phytoremediation of metal-contaminated soils. In this study, we report on the results of a yeast (Saccharomyces cerevisae) complementation screen aimed at identifying candidate heavy metal tolerance genes in T. caerulescens. A number of Thlaspi genes that conferred Cd tolerance to yeast were identified, including possible metal-binding ligands from the metallothionein gene family, and a P-type ATPase that is a member of the P1B subfamily of purported heavy metal-translocating ATPases. A detailed characterization of the Thlaspi heavy metal ATPase, TcHMA4, demonstrated that it mediates yeast metal tolerance via active efflux of a number of different heavy metals (Cd, Zn, lead [Pb], and copper [Cu]) out of the cell. However, in T. caerulescens, based on differences in tissue-specific and metal-responsive expression of this transporter compared with its homolog in Arabidopsis (Arabidopsis thaliana), we suggest that it may not be involved in metal tolerance. Instead, we hypothesize that it may play a role in xylem loading of metals and thus could be a key player in the hyperaccumulation phenotype expressed in T. caerulescens. Additionally, evidence is presented showing that the C terminus of the TcHMA4 protein, which contains numerous possible heavy metal-binding His and Cys repeats residues, participates in heavy metal binding. When partial peptides from this C-terminal domain were expressed in yeast, they conferred an extremely high level of Cd tolerance and Cd hyperaccumulation. The possibilities for enhancing the metal tolerance and phytoremediation potential of higher plants via expression of these metal-binding peptides are also discussed.     

植物修复重金属污染及内生细菌效应

土壤和水体的重金属污染已严重危害人类生存环境与健康。由于受重金属污染的环境分布广泛,迫切需要开发经济的清除环境重金属的技术。植物修复是通过绿色植物降解或移除环境污染物,有望成为重金属污染环境的原位修复技术。植物内生菌是指定殖于健康植物的各种组织和器官内部的细菌,被感染的宿主植物不表现出外在病症,耐重金属的内生菌在多种超富集植物中存在。在植物修复过程中,野生型内生菌或基因工程内生菌的抗性系统能降低重金属植物毒性,促进其迁移金属。耐重金属内生菌还可以通过固氮、溶解矿物元素及产生类植物激素、铁载体和ACC脱氨酶等产物促进植物的生长。主要综述目前植物-内生菌相互作用及其潜在的促进植物修复重金属污染的研究进展。     

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.     

解磷微生物修复土壤重金属污染研究进展

土壤重金属污染问题日益严重,具有普遍性、隐蔽性、表聚性、不可逆性等特点,已经成为环境污染治理中的热点、难点问题。解磷微生物能够依靠自身的代谢产物或通过与其他生物的协同作用,将土壤中的难溶性磷转化为可供植物吸收利用的磷,具有多重植物促生长功能和重金属解毒能力,可在重金属毒害水平下,促进植物生长、提高植物抗病能力、克服重金属对植物生长的不利影响,从而增强重金属修复植物的生存竞争力。从解磷微生物的研究现状入手,介绍了解磷微生物对土壤重金属污染的修复能力,综述了解磷微生物对土壤重金属污染修复的作用机制,分析了目前解磷微生物在重金属修复过程中存在的问题,并提出了今后研究的方向,为重金属污染土壤的修复提供了新思路。     

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.     

Vacuolar compartmentalization: a second-generation approach to engineering plants for phytoremediation

Engineering plants with greater metal tolerance and accumulation properties is the key to developing phytoremediators. A recent study by Won-Yong Song et al. has shown that overexpressing the yeast vacuolar transporter YCF1 increases Pb and Cd tolerance and consequently increases the accumulation of these metals in shoots of transgenic Arabidopsis plants even though expression levels of YCF1 were relatively low. This technology can be used to engineer advanced phytoremediators, increasing their ability to pump heavy metals into a safe compartment while requiring only a small amount of transporters rather than a large amount of chelating peptide material.     

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。