Risk mitigation of genetically modified bacteria and plants designed for bioremediation

While the possible advantages of bioremediation and phytoremediation, by both recombinant microbes and plants, have been extensively reviewed, the biosafety concerns have been less extensively treated. This article reviews the possible risks associated with the use of recombinant bacteria and plants for bioremediation, with particular emphasis on ways in which molecular genetics could contribute to risk mitigation. For example, genetic techniques exist that permit the site-specific excision of unnecessary DNA, so that only the transgenes of interest remain. Other mechanisms exist whereby the recombinant plants or bacteria contain conditional suicide genes that may be activated under certain conditions. These methods act to prevent the spread and survival of the transgenic bacteria or plants in the environment, and to prevent horizontal gene flow to wild or cultivated relatives. Ways in which these genetic technologies may be applied to risk mitigation in bioremediation and phytoremediation are discussed.

     

Expression analysis in response to low temperature stress in blood oranges: implication of the flavonoid biosynthetic pathway

This minireview explores the environmental bioremediation mediated by genetically engineered (GE) bacteria and it also highlights the limitations and challenges associated with the release of engineered bacteria in field conditions. Application of GE bacteria based remediation of various heavy metal pollutants is in the forefront due to eco-friendly and lesser health hazards compared to physico-chemical based strategies, which are less eco-friendly and hazardous to human health. A combination of microbiological and ecological knowledge, biochemical mechanisms and field engineering designs would be an essential element for successful in situ bioremediation of heavy metal contaminated sites using engineered bacteria. Critical research questions pertaining to the development and implementation of GE bacteria for enhanced bioremediation have been identified and poised for possible future research. Genetic engineering of indigenous microflora, well adapted to local environmental conditions, may offer more efficient bioremediation of contaminated sites and making the bioremediation more viable and eco-friendly technology. However, many challenges are to be addressed concerning the release of genetically engineered bacteria in field conditions. There are possible risks associated with the use of GE bacteria in field condition, with particular emphasis on ways in which molecular genetics could contribute to the risk mitigation. Both environmental as well as public health concerns need to be addressed by the molecular biologists. Although bioremediation of heavy metals by using the genetically engineered bacteria has been extensively reviewed in the past also, but the bio-safety assessment and factors of genetic pollution have been never the less ignored.     

有机磷生物修复研究进展

目前,有机磷的生物修复还主要是微生物修复。但是植物修复更具优越性,因其花费更少、对环境更安全。然而植物对生长条件的要求相对较高,修复效率较低,应用还非常有限。本文综述了有机磷微生物修复和植物修复的研究进展,总结了已知的有机磷降解酶及其生物来源。结果表明,植物材料的筛选、土壤与OPs作用机理的研究、植物耐受和消除OPs的基因组学研究、植物-微生物联合降解体系的建立以及降解酶的植物根系分泌系统的利用是提高有机磷植物修复效率的重要途径。     

The use of transgenic plants in the bioremediation of soils contaminated with trace elements

The use of plants to clean-up soils contaminated with trace elements could provide a cheap and sustainable technology for bioremediation. Field trials suggested that the rate of contaminant removal using conventional plants and growth conditions is insufficient. The introduction of novel traits into high biomass plants in a transgenic approach is a promising strategy for the development of effective phytoremediation technologies. This has been exemplified by generating plants able to convert organic and ionic forms of mercury into the less toxic, volatile, elemental mercury, a trait that occurs naturally only in some bacteria and not at all in plants. The engineering of a phytoremediator plant requires the optimization of a number of processes, including trace element mobilization in the soil, uptake into the root, detoxification and allocation within the plant. A number of transgenic plants have been generated in an attempt to modify the tolerance, uptake or homeostasis of trace elements. The phenotypes of these plants provide important insights for the improvement of engineering strategies. A better understanding, both of micronutrient acquisition and homeostasis, and of the genetic, biochemical and physiological basis of metal hyperaccumulation in plants, will be of key importance for the success of phytoremediation.     

Engineering plants for the phytodetoxification of explosives

Summary  Widespread contaimination of the environment by explosives resulting from the manufacture, disposal and testing of munitions is becoming a matter of increasing concern. Most explosives are considered to be a major hazard to biological systems due to their toxic and mutagenic effects. Interest on the bioremediation of land contaminated with explosives has recently been focused on phytoremediation. Unfortunately., whilst plants have many advantages for the remediation of contaminated land and water, they lack the catabolic versatility which enables microorganisms to mineralize such a wide diversity of xenobiotic compounds. This raised the interesting question as to whether the impressive biodegradative capabilities of soil bacteria could be combined with the high biomass and stability of plants to yield an optimal system for in situ bioremediation of explosive residues in soil. Our investigation into the degradation of explosive residues by soil bacteria resulted in the isolation of Enterobacter cloacae PB2, which is capable of utilizing nitrate ester explosives such as pentaerythritol tetranitrate (PETN) and nitroglycerin as the sole source of nitrogen for growth. We have successfully introduced PETN reductase, the enzyme initiating explosive degradation in this organism, into plants to create transgenic plants that degrade explosives. Since the bacterial degradative pathways for many classes of organic pollutant have been elucidated, this may be a generally applicable method of achieving bioremediation of contaminated soil in the environment.     

A comprehensive overview of elements in bioremediation

Sustainable development requires the development and promotion of environmental management and a constant search for green technologies to treat a wide range of aquatic and terrestrial habitats contaminated by increasing anthropogenic activities. Bioremediation is an increasingly popular alternative to conventional methods for treating waste compounds and media with the possibility to degrade contaminants using natural microbial activity mediated by different consortia of microbial strains. Many studies about bioremediation have been reported and the scientific literature has revealed the progressive emergence of various bioremediation techniques. In this review, we discuss the various in situ and ex situ bioremediation techniques and elaborate on the anaerobic digestion technology, phytoremediation, hyperaccumulation, composting and biosorption for their effectiveness in the biotreatment, stabilization and eventually overall remediation of contaminated strata and environments. The review ends with a note on the recent advances genetic engineering and nanotechnology have had in improving bioremediation. Case studies have also been extensively revisited to support the discussions on biosorption of heavy metals, gene probes used in molecular diagnostics, bioremediation studies of contaminants in vadose soils, bioremediation of oil contaminated soils, bioremediation of contaminants from mining sites, air sparging, slurry phase bioremediation, phytoremediation studies for pollutants and heavy metal hyperaccumulators, and vermicomposting.     

金属结合蛋白基因及其在清除重金属污染中的应用

一些微生物和植物由于对毒性金属具有独特的抗性机制,使得利用它们来清除日益严重的环境污染已发展成为一种十分有效的技术——生物修复。研究表明,不同的金属结合蛋白（如MT 和PC）,在生物忍耐和降解过量重金属毒性机制中起重要作用。愈来愈多的MT 和PC基因被克隆,并已成功地应用于生物遗传转化,这些转基因生物在清除重金属污染方面已显示出潜在的应用价值。 Abstract:Heavy metal pollution has become a global environmental hazard.The use of microorganisms and plants for the decontamination of heavy metals is recognized as a low lost and high efficiency method for cleaning up metal contamination.It shows that various metal-binding proteins such as metallothioneins (MTs) or phytochelatines (PCs) play an important role in defense systems and detoxification to heavy metals in organisms.Many genes of MTs and PCs have been cloned and utilized successfully in genetically modified bacteria and plants for increasing remediation capacity.These transgenic organisms have been displayed a great potential in bioremediation and phytoremediation of heavy metals.     

Rhizoremediation: a beneficial plant-microbe interaction

Worldwide, contamination of soil and ground water is a severe problem. The negative effects of pollutants on the environment and on human health are diverse and depend on the nature of the pollution. The search for alternative methods for excavation and incineration to clean polluted sites resulted in the application of bioremediation techniques. In this review, we describe some generally accepted bioremediation tools and subsequently focus on the combination of two approaches, phytoremediation and bioaugmentation, resulting in rhizoremediation. During rhizoremediation, exudates derived from the plant can help to stimulate the survival and action of bacteria, which subsequently results in a more efficient degradation of pollutants. The root system of plants can help to spread bacteria through soil and help to penetrate otherwise impermeable soil layers. The inoculation of pollutant-degrading bacteria on plant seed can be an important additive to improve the efficiency of phytoremediation or bioaugmentation.     

Endophyte-assisted phytoremediation: mechanisms and current application strategies for soil mixed pollutants

Abstract

Phytoremediation uses plants and associated microbes to remove pollutants from the environment and is considered a promising bioremediation method. Compared with well-described single contaminant treatments, the number of studies reporting phytoremediation of soil mixed pollutants has increased recently. Endophytes, including bacteria and fungi, exhibit beneficial traits for the promotion of plant growth, stress alleviation, and biodegradation. Moreover, endophytes either directly or indirectly assist host plants to survive high concentrations of organic and inorganic pollutants in the soil. Endophytic microorganisms can also regulate the plant metabolism in different ways, exhibiting a variety of physiological characteristics. This review summarizes the taxa and physiological properties of endophytic microorganisms that may participate in the detoxification of contaminant mixtures. Furthermore, potential biomolecules that may enhance endophyte mediated phytoremediation are discussed. The practical applications of pollutant-degrading endophytes and current strategies for applying this valuable bio-resource to soil phytoremediation are summarized.     

Endophytic microorganisms—promising applications in bioremediation of greenhouse gases

Bioremediation is a technique that uses microbial metabolism to remove pollutants. Various techniques and strategies of bioremediation (e.g., phytoremediation enhanced by endophytic microorganisms, rhizoremediation) can mainly be used to remove hazardous waste from the biosphere. During the last decade, this specific technique has emerged as a potential cleanup tool only for metal pollutants. This situation has changed recently as a possibility has appeared for bioremediation of other pollutants, for instance, volatile organic compounds, crude oils, and radionuclides. The mechanisms of bioremediation depend on the mobility, solubility, degradability, and bioavailability of contaminants. Biodegradation of pollutions is associated with microbial growth and metabolism, i.e., factors that have an impact on the process. Moreover, these factors have a great influence on degradation. As a result, recognition of natural microbial processes is indispensable for understanding the mechanisms of effective bioremediation. In this review, we have emphasized the occurrence of endophytic microorganisms and colonization of plants by endophytes. In addition, the role of enhanced bioremediation by endophytic bacteria and especially of phytoremediation is presented.     

PAH Phytoremediation: Rhizodegradation or Rhizoattenuation?

Dealing with soil contaminated with persistent organic pollutants (POP) is an increasing concern amplified by both regulatory constraints and the dramatic impact of human activities on the soil resource. The most used management options are treatments which totally eradicate the toxic compounds targeted. When possible, environmental-friendly processes should be used, and recent years have seen the emergence of green technologies using biological energies involving microorganisms (bioremediation) and plants (phytoremediation). Research has focused on phytoremediation and many have presented this technology as the process ideally combining efficiency, low cost and environmental acceptance. However, the applicability of phytoremediation on soils contaminated by bio-recalcitrant organic compounds, such as polycyclic aromatic hydrocarbons (PAH), has not yet proved as successful as expected. We propose here a review and discussion of the overall question of PAH status in soil and their potential for treatment. The limits and applicability of bioremediation technologies are discussed, and the specific beneficial effect of plants is objectively evaluated with a special interest to processes which lead to rhizoattenuation. Given the PAH high affinity to soil organic matter, availability is the main limitation to phytoremediation. In this context, bioavailability quantification remains an issue as well as the characterization of the recalcitrant fraction.     

Using soil bacteria to facilitate phytoremediation

In the past twenty years or so, researchers have endeavored to utilize plants to facilitate the removal of both organic and inorganic contaminants from the environment, especially from soil. These phytoremediation approaches have come a long way in a short time. However, the majority of this work has been done under more controlled laboratory conditions and not in the field. As an adjunct to various phytoremediation strategies and as part of an effort to make this technology more efficacious, a number of scientists have begun to explore the possibility of using various soil bacteria together with plants. These bacteria include biodegradative bacteria, plant growth-promoting bacteria and bacteria that facilitate phytoremediation by other means. An overview of bacterially assisted phytoremediation is provided here for both organic and metallic contaminants, with the intent of providing some insight into how these bacteria aid phytoremediation so that future field studies might be facilitated.     

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.     

Harnessing plant-bacteria-fungi interactions to improve plant growth and degradation of organic pollutants

Exploiting the potential of bacteria in phytoremediation for the removal of organic and inorganic pollutants from soils and (ground)water holds great promise. Besides bacteria, mycorrhizal fungi and free-living saprotrophs are well known for their strong degradative capacities and plant growth promotion effects, which makes them of high interest for use in different bioremediation strategies. To further increase the efficiency and successes of phytoremediation, interactions between plants and their associated microorganisms, both bacteria and fungi, should be further investigated, in addition to the close interactions between bacteria and fungi. Benefitting from an increased understanding of microbial community structure and assembly allows us to better understand how the holobiont can be modified to improve pollutant degradation and plant growth. In this review, we present an overview of insights in plant-bacteria-fungi interactions and the opportunities of exploiting these tripartite interactions to enhance the effectiveness of phytoremediation of organic pollutants.     

Phytoremediation--a novel and promising approach for environmental clean-up

Phytoremediation is an eco friendly approach for remediation of contaminated soil and water using plants. Phytoremediation is comprised of two components, one by the root colonizing microbes and the other by plants themselves, which degrade the toxic compounds to further non-toxic metabolites. Various compounds, viz. organic compounds, xenobiotics, pesticides and heavy metals, are among the contaminants that can be effectively remediated by plants. Plant cell cultures, hairy roots and algae have been studied for their ability to degrade a number of contaminants. They exhibit various enzymatic activities for degradation of xenobiotics, viz. dehalogenation, denitrification leading to breakdown of complex compounds to simple and non-toxic products. Plants and algae also have the ability to hyper accumulate various heavy metals by the action of phytochelatins and metallothioneins forming complexes with heavy metals and translocate them into vacuoles. Molecular cloning and expression of heavy metal accumulator genes and xenobiotic degrading enzyme coding genes resulted in enhanced remediation rates, which will be helpful in making the process for large-scale application to remediate vast areas of contaminated soils. A few companies worldwide are also working on this aspect of bioremediation, mainly by transgenic plants to replace expensive physical or chemical remediation techniques. Selection and testing multiple hyperaccumulator plants, protein engineering ofphytochelatin and membrane transporter genes and their expression would enhance the rate of phytoremediation, making this process a successful one for bioremediation of environmental contamination. Recent years have seen major investments in the R&D, which have also resulted in competition of filing patents by several companies for economic gains. The details of science & technology related to phytoremediation have been discussed with a focus on future trends and prospects of global relevance.     

Role of soil microbes in the rhizospheres of plants growing on trace metal contaminated soils in phytoremediation

This article reviews recent developments in in situ bioremediation of trace metal contaminated soils, with particular reference to the microbial dynamics in the rhizospheres of plants growing on such soils and their significance in phytoremediation. In non-agricultural conditions, the natural role of plant growth promoting rhizobacteria (PGPR), P-solubilizing bacteria, mycorrhizal-helping bacteria (MHB) and arbuscular mycorrhizal fungi (AMF) in maintaining soil fertility is more important than in conventional agriculture, horticulture, and forestry where higher use of agrochemicals minimize their significance. These microbes initiate a concerted action when a particular population density is achieved, i.e. quorum sensing. AMF also recognize their host by signals released by host roots, allowing a functional symbiosis. AM fungi produce an insoluble glycoprotein, glomalin, which sequester trace elements and it should be considered for biostabilization leading to remediation of contaminated soils. Conclusions drawn from studies of metal uptake kinetics in solution cultures may not be valid for more complex field conditions and use of some combination of glasshouse and field experiments with organisms that occur within the same plant community is suggested. Phytoextraction strategies, such as inoculation of plants to be used for phytoremediation with appropriate heavy metal adapted rhizobial microflora, co-cropping system involving a non-mycorrhizal hyperaccumulator plant and a non-accumulator but mycorrhizal with appropriate AMF, or pre-cropping with mycotrophic crop systems to optimize phytoremediation processes, merit further field level investigations. There is also a need to improve our understanding of the mechanisms involved in transfer and mobilization of trace elements by rhizosphere microbiota and to conduct research on selection of microbial isolates from rhizosphere of plants growing on heavy metal contaminated soils for specific restoration programmes. This is necessary if we are to improve the chances of successful phytoremediation.     

Role of soil microbes in the rhizospheres of plants growing on trace metal contaminated soils in phytoremediation

This article reviews recent developments in in situ bioremediation of trace metal contaminated soils, with particular reference to the microbial dynamics in the rhizospheres of plants growing on such soils and their significance in phytoremediation. In non-agricultural conditions, the natural role of plant growth promoting rhizobacteria (PGPR), P-solubilizing bacteria, mycorrhizal-helping bacteria (MHB) and arbuscular mycorrhizal fungi (AMF) in maintaining soil fertility is more important than in conventional agriculture, horticulture, and forestry where higher use of agrochemicals minimize their significance. These microbes initiate a concerted action when a particular population density is achieved, i.e. quorum sensing. AMF also recognize their host by signals released by host roots, allowing a functional symbiosis. AM fungi produce an insoluble glycoprotein, glomalin, which sequester trace elements and it should be considered for biostabilization leading to remediation of contaminated soils. Conclusions drawn from studies of metal uptake kinetics in solution cultures may not be valid for more complex field conditions and use of some combination of glasshouse and field experiments with organisms that occur within the same plant community is suggested. Phytoextraction strategies, such as inoculation of plants to be used for phytoremediation with appropriate heavy metal adapted rhizobial microflora, co-cropping system involving a non-mycorrhizal hyperaccumulator plant and a non-accumulator but mycorrhizal with appropriate AMF, or pre-cropping with mycotrophic crop systems to optimize phytoremediation processes, merit further field level investigations. There is also a need to improve our understanding of the mechanisms involved in transfer and mobilization of trace elements by rhizosphere microbiota and to conduct research on selection of microbial isolates from rhizosphere of plants growing on heavy metal contaminated soils for specific restoration programmes. This is necessary if we are to improve the chances of successful phytoremediation.     

Influence of biochar and compost on phytoremediation of oil-contaminated soil

The use of pyrolyzed carbon, biochar, as a soil amendment is of potential interest for improving phytoremediation of soil that has been contaminated by petroleum hydrocarbons. To examine this question, the research reported here compared the effects of biochar, plants (mesquite tree seedlings), compost and combinations of these treatments on the rate of biodegradation of oil in a contaminated soil and the population size of oil-degrading bacteria. The presence of mesquite plants significantly enhanced oil degradation in all treatments except when biochar was used as the sole amendment without compost. The greatest extent of oil degradation was achieved in soil planted with mesquite and amended with compost (44% of the light hydrocarbon fraction). Most probable number assays showed that biochar generally reduced the population size of the oil-degrading community. The results of this study suggest that biochar addition to petroleum-contaminated soils does not improve the rate of bioremediation. In contrast, the use of plants and compost additions to soil are confirmed as important bioremediation technologies.     

Genomic and evolutionary comparisons of diazotrophic and pathogenic bacteria of the order Rhizobiales

Background

Species belonging to the Rhizobiales are intriguing and extensively researched for including both bacteria with the ability to fix nitrogen when in symbiosis with leguminous plants and pathogenic bacteria to animals and plants. Similarities between the strategies adopted by pathogenic and symbiotic Rhizobiales have been described, as well as high variability related to events of horizontal gene transfer. Although it is well known that chromosomal rearrangements, mutations and horizontal gene transfer influence the dynamics of bacterial genomes, in Rhizobiales, the scenario that determine pathogenic or symbiotic lifestyle are not clear and there are very few studies of comparative genomic between these classes of prokaryotic microorganisms trying to delineate the evolutionary characterization of symbiosis and pathogenesis.

Results

Non-symbiotic nitrogen-fixing bacteria and bacteria involved in bioremediation closer to symbionts and pathogens in study may assist in the origin and ancestry genes and the gene flow occurring in Rhizobiales. The genomic comparisons of 19 species of Rhizobiales, including nitrogen-fixing, bioremediators and pathogens resulted in 33 common clusters to biological nitrogen fixation and pathogenesis, 15 clusters exclusive to all nitrogen-fixing bacteria and bacteria involved in bioremediation, 13 clusters found in only some nitrogen-fixing and bioremediation bacteria, 01 cluster exclusive to some symbionts, and 01 cluster found only in some pathogens analyzed. In BBH performed to all strains studied, 77 common genes were obtained, 17 of which were related to biological nitrogen fixation and pathogenesis. Phylogenetic reconstructions for Fix, Nif, Nod, Vir, and Trb showed possible horizontal gene transfer events, grouping species of different phenotypes.

Conclusions

The presence of symbiotic and virulence genes in both pathogens and symbionts does not seem to be the only determinant factor for lifestyle evolution in these microorganisms, although they may act in common stages of host infection. The phylogenetic analysis for many distinct operons involved in these processes emphasizes the relevance of horizontal gene transfer events in the symbiotic and pathogenic similarity.     

Phytoremediation: synergistic use of plants and bacteria to clean up the environment

Phytoremediation is a relatively new approach to removing contaminants from the environment. It may be defined as the use of plants to remove, destroy or sequester hazardous substances from the environment. Unfortunately, even plants that are relatively tolerant of various environmental contaminants often remain small in the presence of the contaminant. To remedy this situation, plant growth-promoting bacteria that facilitate the proliferation of various plants especially under environmentally stressful conditions may be added to the roots of plants. These bacteria have been selected to lower the level of growth-inhibiting stress ethylene within the plant and also to provide the plant with iron from the soil. The net result of adding these bacteria to plants is a significant increase in both the number of seeds that germinate and the amount of biomass that the plants are able to attain, making phytoremediation in the presence of plant growth-promoting bacteria a much faster and more efficient process.