Micropropagation of <Emphasis Type="Italic">Ailanthus altissima</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis> heavy metal tolerance

Ailanthus altissima, a fast-growing and contamination-resistant species is investigated for its use in areas contaminated by heavy metals. A micropropagation protocol for A. altissima was developed, cultured shoots were tested for in vitro heavy metals tolerance. Proliferation rate and shoot length were affected by 6-benzylaminopurine (BAP) and Murashige and Skoog’s (MS) salt concentrations, best results were obtained in full strength MS medium supplemented with 1.32 or 2.64 μM BAP. Rooting percentage was strongly influenced by indole-3-butyric acid. Cultures of A. altissima exposed to heavy metals demonstrated a tolerance comparable to species already utilized in phytoremediation.     

Expression profiling of <Emphasis Type="Italic">Crambe abyssinica</Emphasis>under arsenate stress identifies genes and gene networks involved in arsenic metabolism and detoxification

Background  

Arsenic contamination is widespread throughout the world and this toxic metalloid is known to cause cancers of organs such as liver, kidney, skin, and lung in human. In spite of a recent surge in arsenic related studies, we are still far from a comprehensive understanding of arsenic uptake, detoxification, and sequestration in plants. Crambe abyssinica, commonly known as 'abyssinian mustard', is a non-food, high biomass oil seed crop that is naturally tolerant to heavy metals. Moreover, it accumulates significantly higher levels of arsenic as compared to other species of the Brassicaceae family. Thus, C. abyssinica has great potential to be utilized as an ideal inedible crop for phytoremediation of heavy metals and metalloids. However, the mechanism of arsenic metabolism in higher plants, including C. abyssinica, remains elusive.     

A plant genetically modified that accumulates Pb is especially promising for phytoremediation

From a number of wild plant species growing on soils highly contaminated by heavy metals in Eastern Spain, Nicotiana glauca R. Graham (shrub tobacco) was selected for biotechnological modification, because it showed the most appropriate properties for phytoremediation. This plant has a wide geographic distribution, is fast-growing with a high biomass, and is repulsive to herbivores. Following Agrobacterium mediated transformation, the induction and overexpression of a wheat gene encoding phytochelatin synthase (TaPCS1) in this particular plant greatly increased its tolerance to metals such as Pb and Cd, developing seedling roots 160% longer than wild type plants. In addition, seedlings of transformed plants grown in mining soils containing high levels of Pb (1572 ppm) accumulated double concentration of this heavy metal than wild type. These results indicate that the transformed N. glauca represents a highly promising new tool for use in phytoremediation efforts.     

Nicotianamine Over-accumulation Confers Resistance to Nickel in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Nicotianamine is a methionine derivative involved in iron homeostasis, able to bind various other metals in vitro. To investigate its role in vivo, we expressed a nicotianamine synthase cDNA (TcNAS1) isolated from the polymetallic hyperaccumulator Thlaspi caerulescens in Arabidopsis thaliana. Transgenic plants expressing TcNAS1 over-accumulated NA, up to 100-fold more than wild type plants. Furthermore, increased NA levels in different transgenic lines were quantitatively correlated with increased nickel tolerance. The tolerance to nickel is expressed at the cellular level in protoplast experiments and is associated with an increased NA content. We have also shown that the most NA-over accumulating line showed a high tolerance to nickel and a significant Ni accumulation in the leaves when grown on nickel-contaminated soil. Our results highlight a new potential role for nicotianamine in heavy metal tolerance at the cellular but also at the whole plant level, easily transposable to a non-tolerant non-hyperaccumulator species. These results open new perspectives for the modulation of nicotianamine content in plants for phytoremediation.     

Overexpression of phytochelatin synthase (<Emphasis Type="Italic">AtPCS</Emphasis>) in rice for tolerance to cadmium stress

Phytoremediation is an important strategy adapted by plants to sequester and/or detoxify pollutants. Phytochelatins, a family of cysteine-rich thiol-reactive peptides, bind to various heavy metals and metalloids making them good candidates for phytoremediation. Phytochelatin synthase catalyses the final step in the biosynthesis of phytochelatins and can be used as a strategy to improve tolerance against heavy metals. In the present study, an AtPCS gene was overexpressed in rice following the in planta transformation approach. Stringent screening strategies were standardized to select putative transformants under a Cd stress of 125 μM at both seedling and plant levels. Molecular analysis by PCR in 18 tolerant plants confirmed the transgene integration and absence of Agrobacterium. Genomic Southern analysis further confirmed the integration of the T-DNA as a single copy. The stability of the T-DNA in the progeny of 5 selected T₁ generation plants was confirmed by tolerance assay, molecular characterization and biochemical analysis for the reduced glutathione, phytochelatin content and lipid peroxidation. This strategy is discussed as a potential mechanism to enhance the tolerance of rice plants to Cd stress.     

Phytoremediation of heavy metal contaminated soil potential by woody plants on Tonglushan ancient copper spoil heap in China

Fast-growing metal-accumulating woody plants are considered potential candidates for phytoremediation of metals. Tonglushan mining, one of the biggest Cu production bases in China, presents an important source of the pollution of environment. The sample was collected at Tonglushan ancient copper spoil heap. The aims were to measure the content of heavy metal in the soil and woody plants and to elucidate the phytoremediation potential of the plants. The result showed that soil Cu, Cd and Pb were the main contamination, the mean contents of which were 3166.73 mg/kg, 3.66 mg/kg and 137.06 mg/kg respectively, which belonged to severe contamination. Fourteen species from 14 genera of 13 families were collected and investigated; except for Ligutrum lucidum, the other 13 woody plants species were newly recorded in this area. In addition, to assess the ability of metal accumulation of these trees, we proposed accumulation index. Data suggested that Platanus × acerilolia, Broussonetia papyrifera, Ligutrum lucidum, Viburnum awabuki, Firmiana simplex, Robina pseudoacacia, Melia azedarach and Osmanthus fragrans exhibited high accumulated capacity and strong tolerance to heavy metals. Therefore, Platanus × acerilolia and Broussonetia papyrifera can be planted in Pb contaminated areas; Viburnum awabuki, Firmiana simplex, Robina pseudoacacia and Melia azedarach are the suitable trees for Cd contaminated areas; Viburnum awabuki, Melia azedarach, Ligutrum lucidum, Firmiana simplex, Osmanthus fragrans and Robina pseudoacacia are appropriate to Cu, Pb and Cd multi-metal contaminated areas.     

Toxicity, accumulation, and removal of heavy metals by three aquatic macrophytes

A comprehensive understanding of the uptake, tolerance, and transport of heavy metals by plants will be essential for the development of phytoremediation technologies. In the present paper, we investigated accumulation, tissue and intracellular localization, and toxic effects of cadmium (Cd), lead (Pb), zinc (Zn), and copper (Cu) in three aquatic macrophytes (the angiosperms Lemna minor and Elodea canadensis, and the moss Leptodictyum riparium). We also tested and compared their capacity to absorb heavy metal from water under laboratory conditions. Our data showed that all the three species examined could be considered good bioaccumulators for the heavy metals tested. L. riparium was the most resistant species and the most effective in accumulating Cu, Zn, and Pb, whereas L. minor was the most effective in accumulating Cd. Cd was the most toxic metal, followed by Pb, Cu, and Zn. At the ultrastructural level, sublethal concentrations of the heavy metals tested caused induced cell plasmolysis and alterations of the chloroplast arrangement. Heavy metal removal experiments revealed that the three macrophytes showed excellent performance in removing the selected metals from the solutions in which they are maintained, thus suggesting that they could be considered good candidates for wastewaters remediation purpose.     

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.     

植物促生菌提高植物重金属耐受性研究进展

近年来植物修复技术因其独特的优势而被广泛关注。许多植物被认为是有价值的利用资源, 然而, 最有实际使用价值的植物对重金属的耐受性有限, 实际应用中变得越来越困难。植物促生菌资源对环境无污染, 具有独特的多样性和巨大的潜力。随着资源的开发和技术的发展, 微生物调控将会使植物修复技术变得更加可行和更有价值。回顾近年来新兴的微生物调控技术, 主要是植物促生菌的筛选、鉴定和应用价值。     

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.     

用转基因植物修复重金属污染的土壤

将新的性状转入高生物量植物中,以此开发高效的转基因植物修复系统,用于重金属污染的土壤修复是一项具有广阔应用前景的技术.大量实验表明,将细菌、真菌、动物、人类及植物本身与重金属脱毒相关的基因转入高生物量植物,异源表达产物可介导转基因植物耐受和高积累重金属及类似物.综述了这方面的研究进展.     

Investigating heavy-metal hyperaccumulation using Thlaspi caerulescens as a model system

Background

Metal-hyperaccumulating plant species are plants that are endemic to metalliferous soils and are able to tolerate and accumulate metals in their above-ground tissues to very high concentrations. One such hyperaccumulator, Thlaspi caerulescens, has been widely studied for its remarkable properties to tolerate toxic levels of zinc (Zn), cadmium (Cd) and sometimes nickel (Ni) in the soil, and accumulate these metals to very high levels in the shoot. The increased awareness regarding metal-hyperaccumulating plants by the plant biology community has helped spur interest in the possible use of plants to remove heavy metals from contaminated soils, a process known as phytoremediation. Hence, there has been a focus on understanding the mechanisms that metal-hyperaccumulator plant species such as Thlaspi caerulescens employ to absorb, detoxify and store metals in order to use this information to develop plants better suited for the phytoremediation of metal-contaminated soils.

Scope

In this review, an overview of the findings from recent research aimed at better understanding the physiological mechanisms of Thlaspi caerulescens heavy-metal hyperaccumulation as well as the underlying molecular and genetic determinants for this trait will be discussed. Progress has been made in understanding some of the fundamental Zn and Cd transport physiology in T. caerulescens. Furthermore, some interesting metal-related genes have been identified and characterized in this plant species, and regulation of the expression of some of these genes may be important for hyperaccumulation.

Conclusions

Thlaspi caerulescens is a fascinating and useful model system not only for studying metal hyperaccumulation, but also for better understanding micronutrient homeostasis and nutrition. Considerable future research is still needed to elucidate the molecular, genetic and physiological bases for the extreme metal tolerance and hyperaccumulation exhibited by plant species such as T. caerulescens.Key words: Zn, Cd, Ni, Thlaspi caerulescens, hyperacumulator, phytoremediation, heavy metal     

用转基因植物修复重金属污染的土壤

将新的性状转入高生物量植物中,以此开发高效的转基因植物修复系统,用于重金属污染的土壤修复是一项具有广阔应用前景的技术。大量实验表明,将细菌、真菌、动物、人类及植物本身与重金属脱毒相关的基因转入高生物量植物,异源表达产物可介导转基因植物耐受和高积累重金属及类似物。综述了这方面的研究进展。     

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.     

Dualities in plant tolerance to pollutants and their uptake and translocation to the upper plant parts

There is a duality in plant tolerance to pollutants and its response to the pollutants’ stress.On the one hand some plants, (hyper)tolerant to heavy metals, are able to hyperaccumulate these metals in shoots, which could be beneficial for phytoremediation purposes to clean-up soil and water. On the other hand tolerant food crops, exposed to heavy metals in their growth medium, may be dangerous as carriers of toxic metals in the food chain leading to food toxicity. There is an additional duality in plant tolerance to heavy metals and that is in food crops that are tolerant and/or hyperaccumulators, which could be used on one hand for phytoremediation, under controlled conditions and on the other hand for food fortification with essential metals.Similarly, plants are also exposed to a large number of xenobiotic organic pollutants. Because they generally cannot avoid these compounds, plants take up, translocate, metabolize and detoxify many of them. There is a large variability in tolerance (defence) mechanisms against organic pollutants among plant species. This includes production of reductants but also scavenger molecules like ascorbate and glutathione and expression of the P-450 defence system, and superfamilies of the enzymes glutathione- and glucosyl-transferases. Again, with view to organic pollutants, plant detoxification mechanisms might well protect the plant itself, but produce compounds with some deleterious potential for other organisms.In this review we discuss these dualities on the basis of examples of agricultural and ‘wild’ species exposed to metal contaminants (mainly Cd) and organic pollutants. Differences in uptake and translocation of various pollutants and their consequences will be considered. We will separately outline the effects of the organic and non-organic pollutants on the internal metabolism and the detoxification mechanisms and try to indicate the differences between both types of pollutants. Finally the consequences and solutions of these dualities in plant tolerance to pollutants will be discussed.     

Genomic pattern of adaptive divergence in Arabidopsis halleri, a model species for tolerance to heavy metal

Pollution by heavy metals is one of the strongest environmental constraints in human-altered environments that only a handful of species can cope with. Identifying the genes conferring to those species the ability to grow in polluted areas is a first step towards a global understanding of the evolutionary processes involved and will eventually improve phytoremediation practices. We used a genome-scan approach to detect loci under divergent selection among four populations of Arabidopsis halleri growing on either polluted or nonpolluted habitats. Based on a high density of amplified fragment length polymorphism (AFLP) markers (820 AFLP markers, i.e. ~1 marker per 0.3 Mb), evidence for selection was found for some markers in every sampled population. Four loci departed from neutrality in both metallicolous populations and thus constitute high-quality candidates for general adaptation to pollution. Interestingly, some candidates differed between the two metallicolous populations, suggesting the possibility that different loci may be involved in adaptation in the different metallicolous populations.     

Use of Cordia Africana in the Phytostabilization of Substrates from Excavations of the Ore Courtyard at the Port of Itaguai,Brazil

The objective of this study was to evaluate the phytostabilization of two substrates contaminated with heavy metals from excavations of the ore courtyard at the port of Itaguai, Brazil. Initially, an inventory of tree species located near the study area was performed to select species for phytostabilization. The species Cordia africana exhibited deeper roots, a larger diameter at chest height (DCH) and crown diameter, as well as higher concentrations of Zn, Cd, and Pb in the trunk, bark, and roots compared to the other investigated species. The tolerance of the selected species to the metals Zn, Cd, and Pb was subsequently assessed through a greenhouse test. The substrates used in the experiment also obtained from excavations at the ore courtyard and showed different levels of heavy metals, indicating either low contamination (Substrate 1) or high contamination (Substrate 2). Alkaline industrial waste steel slag (SS) was used as an amendment to reduce the solubility of heavy metals. Application of the amendment agent to the substrates reduced the bioavailability of heavy metals, favoring the growth of C. africana. This species presents potential for use in phytostabilization programs due to its tolerance for heavy metals and the observed higher accumulation of these metals in the roots and especially the trunk of this species compared to other vegetal parts.     

Isolation and characterization of heavy metal tolerant Gram-positive bacteria with bioremedial properties from municipal waste rich soil of Kestopur canal (Kolkata), West Bengal,India

The present study was conducted to determine the culturable bacterial profile from Kestopur canal (Kolkata, India) and analyze their heavy metal tolerance. In addition to daily sewage including solid and soluble wastes, a considerable load of toxic metals are released into this water body from industries, tanneries and agriculture, household as well as health sectors. Screening out microbes from such an environment was done keeping in mind their multifunctional application especially for bioremediation. Heavy metals are major environmental pollutants when present in high concentration in soil and show potential toxic effects on growth and development in plants and animals. Some edible herbs growing in the canal vicinity, and consumed by people, were found to harbour these heavy metals at sub-toxic levels. The bioconcentration factor of these plants being <1 indicates that they probably only absorb but not accumulate heavy metals. All the thirteen Grampositive bacteria isolated from these plants rhizosphere were found to tolerate high concentration of heavy metals like Co, Ni, Pb, Cr, Fe. Phylogenetic analysis of their 16S rDNA genes revealed that they belonged to one main taxonomic group — the Firmicutes. Seven of them were found to be novel with 92–95% sequence homology with known bacterial strains. Further microbiological analyses show that the alkaliphilic Bacillus weihenstephanensis strain IA1 and Exiguobacterium aestuarii strain CE1, with selective antibiotic sensitivity along with high Ni²⁺ and Cr⁶⁺ removal capabilities, respectively, can be prospective candidates for bioremediation.