Transgenic plants in phytoremediation: recent advances and new possibilities

Phytoremediation, the use of plants and their associated microbes to remedy contaminated soils, sediments, and groundwater, is emerging as a cost-effective and environmentally friendly technology. Due in large part to its aesthetic appeal, this technology has gained increasing attention over the past 10 years. Phytoremediation uses different plant processes and mechanisms normally involved in the accumulation, complexation, volatilization, and degradation of organic and inorganic pollutants. Certain plants, called hyperaccumulators, are good candidates in phytoremediation, particularly for the removal of heavy metals. Phytoremediation efficiency of plants can be substantially improved using genetic engineering technologies. Recent research results, including overexpression of genes whose protein products are involved in metal uptake, transport, and sequestration, or act as enzymes involved in the degradation of hazardous organics, have opened up new possibilities in phytoremediation. This paper provides a critical review of the recent progress made toward the development of transgenic plants with improved phytoremediation capabilities and their potential use in environmental cleanup.     

Secretion of bacterial xenobiotic-degrading enzymes from transgenic plants by an apoplastic expressional system: an applicability for phytoremediation

In search of an effective method for phytoremediation of wastewater contaminated with organic compounds, we investigated the application of an apoplastic expressional system that secretes useful bacterial enzymes from transgenic plants into hydroponic media through the addition of a targeting signal. We constructed transgenic Arabidopsis expressing the aromatic-cleaving extradiol dioxygenase (DbfB), which degrades 2,3-dihydroxybiphenyl (2,3-DHB), and transgenic tobacco expressing haloalkane dehalogenase (DhaA), which catalyzes hydrolytic dechlorination of 1-chlorobutane (1-CB). Although crude leaf extracts of transgenic plants expressing cytoplasm-targeted degradative enzymes showed higher activity than did those from transgenic plants expressing apoplast-targeted enzymes, the hydroponic media of the latter showed 23.2 times (DbfB) and 76.4 times (DhaA) higher activity than plants containing the cytoplasm-targeted enzymes. Addition of crystalline 2,3-DHB to 100 mL of the hydroponic medium of transgenic or wild-type seedlings revealed that only medium from the transgenic Arabidopsis expressing apoplast-targeted DbfB showed rapid ring cleavage of 2,3-DHB. Transgenic tobacco expressing apoplast-targeted DhaA also resulted in the accumulation of the dehalogenation product 1-butanol in the hydroponic medium and showed a higher tolerance to 1-CB than wild-type or transgenic plants expressing cytoplasm-targeted DhaA. These results demonstrate the usefulness of the apoplastic expression of bacterial recombinant proteins in phytoremediation.     

野苋菜植物修复皮革工业铬污染土壤的研究

为选择超积累重金属和对气候具有耐性的植物,以达到利用植物修复重金属污染土壤的目的。采用田间种植,比较了超积累重金属植物大叶红苋菜(298)与野苋菜、向日葵、玉米等4种植物修复铬污染。根茎叶经晒干灼烧,回收铬处理,而实现无二次污染修复土壤。研究结果表明:这些植物的根茎叶对皮革工业铬污染具有较强的吸收和积累能力,野苋菜的地上部生物量分别是向日葵的1.5倍、玉米的1.2倍、大叶红苋菜(298)的1.05倍。对于地下部生物量而言,4种植物的高低顺序依次为野苋菜>大叶红苋菜(298)>玉米>向日葵。4种植物的根茎叶对铬累积量的高低顺序依次为野苋菜>大叶红苋菜(298)>向日葵>玉米。野苋菜能吸收富集铬重金属并且具有耐铬重金属的特性,基于其具有生物量较大、生长速度快、耐高温、耐干旱的特点,作为重金属污染的修复植物具有较好的应用前景。     

Natural revegetation in the vicinity of the former lead smelter in Zerjav, Slovenia

The response of plant communities to pollution associated with the lead smelter in Zerjav, Slovenia, was investigated on spatial and temporal scales. In 2001, the total concentrations of contaminating metals in the soil measured at the most polluted plot were 59000 mg kg(-1) Pb, 180 mg kg(-1) Cd, and 3300 mg kg(-1) Zn. A negative correlation between the concentration of metals in the soil and plant biodiversity parameters along the gradient of pollution in 2001 was detected. Plant species lists were compiled in 2001 for plots located at different distances from the emission source and compared to that of 1981. In the period from 1981 to 2001, smelter emissions were reduced, and plant species richness increased at all examined plots. Among the successful survivals were some metal hyperaccumulators (Minuartia gerardii, Thlaspi praecox, and Biscutella laevigata). Of special interest were plants that survived the period of highest pollution. We believe that these species can be used in metal-degraded environments for natural revegetation to immobilize heavy metals. The ecosystem in the surroundings of the former smelter is presently recovering. Our results suggest that high metal concentrations in soil are a potential limiting factor for revegetation.     

Insights into the structural and functional evolution of plant genomes afforded by the nucleotide sequences of chromosomes 2 and 4 of Arabidopsis thaliana

The rapidly accumulating genome sequence data from the plant Arabidopsis thaliana allows more detailed analysis of genome content and organisation than ever before possible in plants. The genome shows a surprisingly high level of genetic redundancy, with as many as 75% of gene products showing significant homology to another protein of A. thaliana. Many duplicated genes occur in arrays of conserved order and indicate that A. thaliana is likely to have had a tetraploid ancestor. Analysis of the divergence of duplicated genome segments leads to the prediction of two major modes of plant genome evolution: macro-scale duplication and rearrangement of chromosomes and micro-scale translocations, duplication and loss of individual genes or small groups of genes.     

Identification of putative gene encoded on ORF16 of the 81 kb contig of Arabidopsis thaliana chromosome III as alpha-mannosidase

We isolated cDNA corresponding to open reading frame (ORF) 16 of the 81 kb contig of Arabidopsis thaliana chromosome III [Quigley., Nucleic Acids Res., 24, 4313-4318 (1996)] and expressed alpha-mannosidase activity in tobacco suspension-cultured cells, which revealed that ORF16 encodes alpha-mannosidase. We also suggested that Arabidopsis harbors three genes encoding alpha-mannosidase by homology search against the database.     

重金属污染对葡萄生化及安全品质影响的研究进展

对国内外有关重金属对葡萄生理生化、安全品质影响及缓解措施的最新研究进展进行归纳总结,旨在为相关领域研究人员系统了解葡萄对重金属污染的响应及其缓解机理提供参考。重金属污染对葡萄根系和茎生长具有抑制作用,影响氮、铁等营养元素吸收,并能使葡萄叶片卷曲,抑制光合作用。重金属胁迫会引起葡萄氧化胁迫,比如膜脂过氧化和蛋白质损伤等,进而导致植物代谢紊乱,甚至植株死亡。重金属在葡萄植株中积累因其种类和葡萄品种不同而不同,总体而言,葡萄根、茎、叶相对于葡萄果实对重金属的富集能力更强。在重金属污染的葡萄园内施用含硅材料和有机物类肥料,可以起到缓解重金属毒害,提升土壤环境质量的作用。     

Accumulation of Cu, Pb, Ni and Zn in the halophyte plant Atriplex grown on polluted soil

BACKGROUND: Three annual Atriplex species—A. hortensis var. purpurea, A. hortensis var. rubra and A. rosea—growing on soil with various levels of the heavy metals copper, lead, nickel, and zinc, have been investigated. RESULTS: Metal accumulation by Atriplex plants differed among species, levels of polluted soil and tissues. Metals accumulated by Atriplex were mostly distributed in root tissues, suggesting that an exclusion strategy for metal tolerance widely exists in them. The increased concentration of heavy metals in soil led to increases in heavy metal shoot and root concentrations of Ni, Cu, Pb and Zn in plants as compared to those grown on unpolluted soil. Accumulation was higher in roots than shoots for all the heavy metals. None of the plants were suitable for phytoextraction because no hyperaccumulator was identified. However, plants with a high bioconcentration factor and low translocation factor have the potential for phytostabilization. Similarly, the correlation between metal concentrations and translocations in plants (BCFs and TFs) using a linear regression was also statistically significant. CONCLUSION: Among the plants studied, var. purpurea was the most efficient in accumulating Pb and Zn in its shoots, whereas var. rubra was most suitable for phytostabilization of sites contaminated with Cu and Ni. Copyright © 2011 Society of Chemical Industry     

Enantioselective Damage of Diclofop Acid Mediated by Oxidative Stress and Acetyl-CoA Carboxylase in Nontarget Plant Arabidopsis thaliana

Diclofop-methyl (DM) is a widely used chiral herbicide, which rapidly hydrolyzes to its major metabolite diclofop acid (DC) after application. With a carbon chiral center, DC not only is an important ingredient of herbicidal activity, but also has a long half-life in soil. Studies so far have only considered the activity of racemic DM in target organisms, and the enantioselective toxicity in nontarget plants of DM and DC has yet to be explored. In this study, the enantioselective phytotoxicity of DC mediated by oxidative stress and the key enzyme ACCase in the fatty acid synthesis system on the model plant Arabidopsis thaliana was investigated. Significant differences between the two enantiomers were observed in phytotoxicity including growth inhibition, oxidative damage and alteration of key genes expression of ACCase, with R-DC showing greater toxicity to Arabidopsis thaliana than S-DC. The results of molecular docking showed that there was a stronger affinity between R-DC and the target enzyme carboxyltransferase domain of ACCase, likely leading to the enantioselective phytotoxicity of DC. This study suggested that chirality of both parent compounds and metabolites should be considered to improve our understanding of the environmental fate and risks of chiral pesticides.     

人工microRNAs干扰At3A06基因增强了拟南芥对菌核病的敏感性

基于本研究室自主开发的油菜防御cDNA芯片分析结果,油菜基因Bn3A06受菌核病诱导后特异上调表达。序列比对发现,该基因与拟南芥基因At3A06高度同源,后者编码289个氨基酸,功能未知。本研究用花序浸染法将At3A06基因的人工微RNA（amiRNA）干扰载体（该载体包含一个除草剂草胺磷抗性基因）转化拟南芥。经草胺磷筛选获得3株T1代转基因植株,PCR检测均为阳性;荧光定量PCR检测表明,与野生型对照相比,3个转基因植株的At3A06基因的表达量均显著降低;对其T2代进行菌核病抗性鉴定,结果显示转基因株系对菌核病的敏感性均显著增强（P〈0.05）。推测At3A06基因可能参与了植物的防御反应,与植株对核盘菌抗性相关。     

A facile transformation of Arabidopsis thaliana using ceramic supported propagation system

Arabidopsis thaliana can be a useful model plant in the field of plant molecular biology because it is a sole dicotyledonous plant whose genome sequence has been completely determined at the present time. To develop functional genomics of A. thaliana, a large number of transgenic plants must be produced. However, transformation of the plants requires wide space and long time. For these reasons, a highly efficient method has been desired to be developed. In order to save space, the transformation system was improved, based on a new technology for plant propagation using ceramic tubes. The new method using ceramic tubes facilitated to prepare plants with a uniform growth level. In addition, the colander procedure after infiltration, which is one of tedious procedures, could be omitted. The new system without soil usage could cut down the cleaning up after experiment.     

Impact of transgenic tobacco on trinitrotoluene (TNT) contaminated soil community

Environmental contamination with recalcitrant toxic chemicals presents a serious and widespread problem to the functional capacity of soil. Soil bacteria play an essential role in ecosystem processes, such as nutrient cycling and decomposition; thus a decrease in their biomass and community diversity, resulting from exposure to toxic chemicals, negatively affects the functioning of soil. Plants provide the primary energy source to soil microorganisms and affect the size and composition of microbial communities, which in turn have an effect on vegetation dynamics. We have found that transgenic tobacco plants overexpressing a bacterial nitroreductase gene detoxify soil contaminated with the high explosive 2,4,6-trinitrotoluene (TNT), with a significantly increased microbial community biomass and metabolic activity in the rhizosphere of transgenic plants compared with wild type plants. This is the first report to demonstrate that transgenic plants engineered for the phytoremediation of organic pollutants can increase the functional and genetic diversity of the rhizosphere bacterial community in acutely polluted soil compared to wild type plants.     

Application of microbial genes to recalcitrant biomass utilization and environmental conservation

Recent papers concerning the application of microbial genes to recalcitrant biomass utilization and environmental conservation are reviewed. Microbial genes have been integrated and expressed in plants and microorganisms. When cellulose-degrading enzyme genes are expressed in rice plants, the transgenic plants exhibit swollen cell walls which increases the digestibility of rice straw in the rumen. When genes encoding aromatic compound-degrading enzymes are expressed in plants, it is expected that aromatic compounds contaminating soil would be degraded during the growth of the transgenic plants. The former transgenic plants are utilized as feed and the latter for phytoremediation. Dockerin and cohesin interactions occurring in the cellulase complex, cellulosome, are applied to the construction of artificial enzyme complexes and protein purification by expressing the genes in transformed bacteria and/or silkworms, respectively. In the case of the forced expression of bacterial genes encoding chitinase and/or hydrogenase in the wild-type bacteria, chitin degradation and hydrogen gas production in the transformed bacteria occur at much higher rates than in the wild type.     

农杆菌介导microRNA398靶基因转化大豆

拟南芥miR398对低磷胁迫有响应。通过生物信息学方法预测找到大豆miR398a的靶基因铜/锌超氧化物歧化酶GmSODC（Copper/zinc superoxide dismutase）,通过RT-PCR扩增得到该酶的全长cDNA,采用LIC（Ligation-Independent cloning）法将GmSODC连接到双元植物表达载体pJG045上,构建成植物表达载体pSDC6。通过农杆菌介导的子叶节法将GmSODC基因导入大豆品种东农50中,获得了转化GmSODC的T1植株。采用Real-time PCR方法,对转基因植株进行GmSODC基因转录水平的相对定量分析,其中1株较非转基因植株表达量明显提高。     

重金属和抗生素抗性基因在土壤中的复合污染研究进展

抗生素滥用及其诱导产生的抗生素抗性基因(antibiotic resistance genes, ARGs)污染己经引起人们的广泛关注。与此同时,环境中重金属的污染状况也呈加剧趋势,与抗生素抗性基因不同,重金属在自然环境中很难被降解,二者同时存在不仅维持和加强了环境中的微生物对重金属的原有抗性,同时增强了对抗生素抗性基因的抗性。目前,重金属污染地区的养殖业中动植物细菌传染病的抗生素治疗总是无效的,其原因可能是重金属离子和抗生素抗性基因的协同抗性在选择压力下得到增强。本文总结了我国抗生素抗性基因和重金属元素在土壤中的污染状况及其复合污染特征,综述了这2类物质在土壤中的协同选择机制和综合修复措施,以期为重金属和抗生素抗性基因带来的污染所致危害的减轻和未来复合污染相关研究的深入提供参考。     

重金属污染土壤固化/稳定化修复技术的研究

重金属污染土壤的修复问题一直是国内外研究课题的难点.目前,对此类污染土壤的处理主要包括物理、化学、生物等修复方法,其中,物理法主要包括:客土法、换土法和深耕翻土法;而化学法主要包括:固定/稳定化技术、土壤淋洗技术和氧化还原技术;生物法主要包括植物修复和生物治理.基于以上方法,在重金属污染土壤的治理修复过程中,最常用的方...     

Fractionation of stable iron isotopes in higher plants

Although the fractionation of stable iron isotopes by biological processes in the environment is currently a matter of intense debate, the isotope fractionation associated with the growth of higher plants has, to date, not been characterized. We show that iron isotope fractionation induced by higher plants is substantial and also generates systematic plant-specific patterns. We suggest a hypothesis in which these patterns mirror the two different strategies that plants have developed to incorporate iron from the soil: reduction of Fe(III) in soils by strategy I plants results in the uptake of iron, which is depleted in 56Fe by up to 1.6 per mil relative to 54Fe when compared to the available Fe in soils; complexation with siderophores by strategy II plants results in the uptake of iron that is 0.2 per mil heavier than that in soils. Furthermore, younger parts of strategy I plants get increasingly depleted in heavy isotopes as the plant grows, while strategy II plants incorporate nearly the same isotope composition throughout. This points to entirely different translocation mechanisms between strategy I and II plants. Such presumably redox-related differences in translocation have been under debate up to now. We conclude that plant metabolism represents an important cause of isotopic variation in the biogeochemical cycling of Fe. Therefore, heavy stable metal isotope systems now start to be viable indicators of geosphere-biosphere metal transfer processes.