Genetic improvement of heavy metal tolerance in plants by transfer of the yeast metallothionein gene (CUP1)

Cauliflower (Brassica oleracea var. botrytis) tolerates treatment with 25 µM CdCl2 for eight days, but is killed by that with a 50 µM concentration. However, even 15 µM CdCl2 is toxic in the presence of 1 mM L-buthionine sulfoximine (BSO), suggesting the presence of a Cd-inducible phytochelatin and its involvement in Cd-tolerance in cauliflower. To develop heavy metal-tolerant transgenic plants, we ligated the structural gene of yeast metallothionein gene (CUP1) downstream of CaMV35S promoter and introduced the fused gene into cauliflower. A Cd-tolerant transgenic cauliflower was selected, which grew well in the presence of 400 µM or less Cd, whereas the non-transformed cauliflower tolerated only up to 25 µM Cd. The transgenic cauliflower accumulated more Cd, especially in the upper leaves, than the non-transformed plant.In conclusion, by transfer of the yeast metallothionein gene into cauliflower increased Cd-tolerance and Cd-accumulating ability can be conferred to the plant.     

Heavy metal tolerance of transgenic tobacco plants over-expressing cysteine synthase

Cysteine synthase [O-acetyl-L-serine(thiol)lyase] catalyzes the final step for L-cysteine biosynthesis in plants. The tolerance of transgenic tobacco plants over-expressing cysteine synthase cDNA in cytosol (3F), chloroplasts (4F) and in both organelles (F1) was investigated towards heavy metals such as Cd, Se, Ni, Pb and Cu. The transgenic plants were significantly more tolerant than wild-type plants in agar medium containing Cd, Se and Ni. The F1 transgenic plants had a higher resistance than other transgenic lines towards these metals and could enhance accumulation of Cd in shoot. These results suggest that the transgenic plants over-expressing cysteine synthase both in cytosol and chloroplasts can be applicable to phyto-remediation of Cd from contaminated soils.     

Phytochelatins as biomarkers for heavy metal stress in maize (Zea mays L.) and wheat (Triticum aestivum L.): combined effects of copper and cadmium

Heavy metal contaminated soils often show increased levels of more than one metal, e.g. copper (Cu), cadmium (Cd), zinc (Zn), lead (Pb) or nickel (Ni). In case such soils are used for crop production, prediction of yield reduction or quality decline due to heavy metals in the soil is inadequate when based only on chemical soil analysis. The use of biomarkers such as phytochelatins (PC), non-protein thiols specifically induced in plants upon exposure to heavy metals, may be an additional tool or diagnostic criterion in heavy metal research and in practice. In the present work, Cu and Cd uptake and induction of PC synthesis are studied with hydroponically grown maize and wheat plants exposed to mixtures of the two metals. We observed a close positive relationship between the concentrations of Cd and PC in the plant shoot material. A decreased shoot concentration of Cd after addition of Cu, due to metal competition at common root absorption sites, coincided with lower shoot PC levels. Also differences in metal uptake and xylary metal transport among the two plant species were reflected in corresponding differences in PC concentration. The observed direct relationship between shoot PC concentration and the degree of metal-induced growth inhibition makes the use of PC promising for the purpose tested for.     

超富集植物对重金属耐受和富集机制的研究进展

超富集植物对重金属耐受和富集机制的研究成为近年来植物逆境生理研究的热点,在简要总结细胞壁沉淀、重金属螯合效应、酶活性机制和细胞区室化作用的基础上,概述了超富集植物对重金属的耐受机制,讨论了重金属跨根细胞质膜运输,共质体内运输、木质部运输和跨叶细胞膜运输的富集过程。     

重金属污染土壤的湿地生物修复技术

生物修复技术是近年来发展起来的一种有前途的污染治理技术,重金属的湿地植物稳定技术和植物萃取技术等植物修复技术是一研究热点;同时,针对微生物对重金属的生物积累、生物转化及生物修复作一分析,重金属的生物修复生理机制及其提高富集效率的条件作一综述,以期推动国内这一国际热点领域的研究。     

植物螯合肽及其在抗重金属胁迫中的作用

植物螯合肽(PCs)广泛存在于植物体中,与植物抗重金属胁迫关系密切。植物螯合肽及其复合物是一类富含半胱氨酸的低分子量化合物。现有研究证明,PCS由谷胱甘肽(GSH)为底物的酶促反应合成,其合成受相关基因的调控,从模式植物拟南芥的突变体中已分离到与PCS合成有关的几个基因。植物螯合肽首先与重金属离子结合形成低分子量(LMW)复合物,以此形态经由细胞质进入液泡后,再与一个分子的植物螯合肽结合,形成对植物组织毒性较小的高分子量(HMW)复合物,从而达到缓解重金属对植物的危害作用。就植物螯合肽及其复合物的结构、生物合成、基因调控及重金属解毒机理等进行了综述,并对今后的研究方向提出了一些看法。     

The phytochelatin transporters AtABCC1 and AtABCC2 mediate tolerance to cadmium and mercury

Heavy metals such as cadmium (Cd) and mercury (Hg) are toxic pollutants that are detrimental to living organisms. Plants employ a two-step mechanism to detoxify toxic ions. First, phytochelatins bind to the toxic ion, and then the metal-phytochelatin complex is sequestered in the vacuole. Two ABCC-type transporters, AtABCC1 and AtABCC2, that play a key role in arsenic detoxification, have recently been identified in Arabidopsis thaliana. However, it is unclear whether these transporters are also implicated in phytochelatin-dependent detoxification of other heavy metals such as Cd(II) and Hg(II). Here, we show that atabcc1 single or atabcc1 atabcc2 double knockout mutants exhibit a hypersensitive phenotype in the presence of Cd(II) and Hg(II). Microscopic analysis using a Cd-sensitive probe revealed that Cd is mostly located in the cytosol of protoplasts of the double mutant, whereas it occurs mainly in the vacuole of wild-type cells. This suggests that the two ABCC transporters are important for vacuolar sequestration of Cd. Heterologous expression of the transporters in Saccharomyces cerevisiae confirmed their role in heavy metal tolerance. Over-expression of AtABCC1 in Arabidopsis resulted in enhanced Cd(II) tolerance and accumulation. Together, these results demonstrate that AtABCC1 and AtABCC2 are important vacuolar transporters that confer tolerance to cadmium and mercury, in addition to their role in arsenic detoxification. These transporters provide useful tools for genetic engineering of plants with enhanced metal tolerance and accumulation, which are desirable characteristics for phytoremediation.     

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。     

一株具有植物促生作用的重金属铅吸附菌的筛选与鉴定

【背景】随着工业化的发展,重金属污染逐渐成为主要的环境污染之一。微生物修复去除重金属污染成为近些年来新兴的修复方法,筛选开发具有良好修复功能的微生物菌株具有重要的现实意义。【目的】筛选具有促进植物生长作用的重金属修复菌株,为生物修复和植物促生等综合开发利用提供微生物资源。【方法】利用选择性培养基从淤泥中筛选重金属铅的抗性菌株,根据形态学观察、生理生化鉴定和16S rRNA基因序列分析对菌株进行分离鉴定,通过单因素分析不同培养条件对菌株生长的影响;采用原子吸收光谱法、比色法及平板对峙法等对菌株的重金属铅吸附率、无机磷溶解能力、吲哚乙酸(indole-3-acetic acid,IAA)分泌及拮抗镰刀菌效果等进行分析。【结果】从污染严重的塘泥中筛选到一株对重金属铅有较好吸附率的菌株,在150 mg/L Pb²⁺浓度下,对Pb²⁺的吸附率达90%以上;初步鉴定该菌株为蜡样芽孢杆菌,命名为SEM-15;菌株还具有较好的溶解无机磷、分泌IAA及拮抗镰刀菌的能力;菌株生长适应性强,可以在pH 10.0的强碱性环境下生长,该菌株具有很好的重金属铅污染修复及促生防病的应用潜力。【结论】菌株SEM-15是一株具有植物促生作用的重金属铅吸附菌株,在重金属污染土壤联合植物修复的应用中可能具有较好的开发价值。     

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。     

Chemical and enzymatic extraction of heavy metal binding polymers from isolated cell walls of Saccharomyces cerevisiae

Isolated cell walls of the yeast Saccharomyces cerevisiae were treated by either chemical (alkali and acid) or enzymatic (protease, mannanase or beta-glucuronidase) processes to yield partially purified products. These products were partially characterized by infrared analysis. They were subsequently reacted with heavy metal cation solutions and the quantity of metal accumulated by the cell wall material determined. The Cu(2+) ion (0.24, 0.36, 1.12, and 0.60 mumol/mg) was accumulated to a greater extent than either Co(2+) (0.13, 0.32, 0.43, and 0.32 mumol/mg) or Cd(2+) (0.17, 0.34, 0.39, and 0.32 mumol/mg) by yeast cell walls, glucan, mannan, and chitin, respectively The isolated components each accumulated greater quantities of the cations than the intact cell wall. Removal of the protein component of the yeast cell walls by Pronase caused a 29.5% decrease in metal accumulation by yeast cell walls per mass, indicating the protein is a heavy metal accumulating component. The data indicate that the outer mannan-protein layer of the yeast cell wall is more important than the inner glucan-chitin layer in heavy metal action accumulation. (c) 1994 John Wiley & Sons, Inc.     

The role of the root cell wall in the heavy metal tolerance ofAthyrium yokoscense

Cells of the roots ofA. yokoscense growing on metalliferous habitats were fractionated into their cell wall and cytoplasmic components. About 70–90% of the total copper, zinc and cadmium was located in the cell wall. Copper had a markedly greater affinity for the cell wall than zinc and cadmium, and was prevented from entering the cytoplasm. A large proportion of these heavy metals in the cell wall were exchanged as ions. The capacity of the cell wall for exchanging metal ions inA. yokoscense was higher than in other plants growing on metalliferous habitats. However, compared with different ferns unable to grow on metalliferous habitats, this capacity was not unique toA. yokoscense. Consequetly, the root cell wall ofA. yokoscense is considered to be an important site of metal ion storage and may play the role of an excretory organ for heavy metals. On the other hand, as proportion of the heavy metls was transported to the cytoplasm, where the metal content was much higher than the average for normal ferns. This would suggest thatA. yokoscense has another metabolic mechanism related to metal tolerance.     

Physiological and molecular mechanisms of heavy metal accumulation in nonmycorrhizal versus mycorrhizal plants

Uptake, translocation, detoxification, and sequestration of heavy metals (HMs) are key processes in plants to deal with excess amounts of HM. Under natural conditions, plant roots often establish ecto‐ and/or arbuscular‐mycorrhizae with their fungal partners, thereby altering HM accumulation in host plants. This review considers the progress in understanding the physiological and molecular mechanisms involved in HM accumulation in nonmycorrhizal versus mycorrhizal plants. In nonmycorrhizal plants, HM ions in the cells can be detoxified with the aid of several chelators. Furthermore, HMs can be sequestered in cell walls, vacuoles, and the Golgi apparatus of plants. The uptake and translocation of HMs are mediated by members of ZIPs, NRAMPs, and HMAs, and HM detoxification and sequestration are mainly modulated by members of ABCs and MTPs in nonmycorrhizal plants. Mycorrhizal‐induced changes in HM accumulation in plants are mainly due to HM sequestration by fungal partners and improvements in the nutritional and antioxidative status of host plants. Furthermore, mycorrhizal fungi can trigger the differential expression of genes involved in HM accumulation in both partners. Understanding the molecular mechanisms that underlie HM accumulation in mycorrhizal plants is crucial for the utilization of fungi and their host plants to remediate HM‐contaminated soils.     

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.     

水体重金属污染植物修复研究进展

植物修复技术作为一种废水处理新技术,具有投资少、效果好、产出高、环境效益好等优点,现已得到迅速发展及广泛应用。介绍了水生超富集植物的3种生态类型,造成水体污染重金属元素类型,植物应答重金属污染的检测方法,并对植物修复水体重金属污染的发展前景提出展望。     

Phytochelatin–metal(loid) transport into vacuoles shows different substrate preferences in barley and Arabidopsis

Cadmium (Cd) and arsenic (As) are toxic to all living organisms, including plants and humans. In plants, Cd and As are detoxified by phytochelatins (PCs) and metal(loid)‐chelating peptides and by sequestering PC–metal(loid) complexes in vacuoles. Consistent differences have been observed between As and Cd detoxification. Whereas chelation of Cd by PCs is largely sufficient to detoxify Cd, As–PC complexes must be sequestered into vacuoles to be fully detoxified. It is not clear whether this difference in detoxification pathways is ubiquitous among plants or varies across species. Here, we have conducted a PC transport study using vacuoles isolated from Arabidopsis and barley. Arabidopsis vacuoles accumulated low levels of PC₂–Cd, and vesicles from yeast cells expressing either AtABCC1 or AtABCC2 exhibited negligible PC₂–Cd transport activity compared with PC₂–As. In contrast, barley vacuoles readily accumulated comparable levels of PC₂–Cd and PC₂–As. PC transport in barley vacuoles was inhibited by vanadate, but not by ammonium, suggesting the involvement of ABC‐type transporters. Interestingly, barley vacuoles exhibited enhanced PC₂ transport activity when essential metal ions, such as Zn(II), Cu(II) and Mn(II), were added to the transport assay, suggesting that PCs might contribute to the homeostasis of essential metals and detoxification of non‐essential toxic metal(loid)s.