Rhizosphere processes and management in plant-assisted bioremediation (phytoremediation) of soils

Plant-assisted bioremediation or phytoremediation holds promise for in situ treatment of polluted soils. Enhancement of phytoremediation processes requires a sound understanding of the complex interactions in the rhizosphere. Evaluation of the current literature suggests that pollutant bioavailability in the rhizosphere of phytoremediation crops is decisive for designing phytoremediation technologies with improved, predictable remedial success. For phytoextraction, emphasis should be put on improved characterisation of the bioavailable metal pools and the kinetics of resupply from less available fractions to support decision making on the applicability of this technology to a given site. Limited pollutant bioavailability may be overcome by the design of plant–microbial consortia that are capable of mobilising metals/metalloids by modification of rhizosphere pH (e.g. by using Alnus sp. as co-cropping component) and ligand exudation, or enhancing bioavailability of organic pollutants by the release of biosurfactants. Apart from limited pollutant bioavailability, the lack of competitiveness of inoculated microbial strains (in particular degraders) in field conditions appears to be another major obstacle. Selecting/engineering of plant–microbial pairs where the competitiveness of the microbial partner is enhanced through a “nutritional bias” caused by exudates exclusively or primarily available to this partner (as known from the “opine concept”) may open new horizons for rhizodegradation of organically polluted soils. The complexity and heterogeneity of multiply polluted “real world” soils will require the design of integrated approaches of rhizosphere management, e.g. by combining co-cropping of phytoextraction and rhizodegradation crops, inoculation of microorganisms and soil management. An improved understanding of the rhizosphere will help to translate the results of simplified bench scale and pot experiments to the full complexity and heterogeneity of field applications.     

Microbial expression profiles in the rhizosphere of willows depend on soil contamination

The goal of phytoremediation is to use plants to immobilize, extract or degrade organic and inorganic pollutants. In the case of organic contaminants, plants essentially act indirectly through the stimulation of rhizosphere microorganisms. A detailed understanding of the effect plants have on the activities of rhizosphere microorganisms could help optimize phytoremediation systems and enhance their use. In this study, willows were planted in contaminated and non-contaminated soils in a greenhouse, and the active microbial communities and the expression of functional genes in the rhizosphere and bulk soil were compared. Ion Torrent sequencing of 16S rRNA and Illumina sequencing of mRNA were performed. Genes related to carbon and amino-acid uptake and utilization were upregulated in the willow rhizosphere, providing indirect evidence of the compositional content of the root exudates. Related to this increased nutrient input, several microbial taxa showed a significant increase in activity in the rhizosphere. The extent of the rhizosphere stimulation varied markedly with soil contamination levels. The combined selective pressure of contaminants and rhizosphere resulted in higher expression of genes related to competition (antibiotic resistance and biofilm formation) in the contaminated rhizosphere. Genes related to hydrocarbon degradation were generally more expressed in contaminated soils, but the exact complement of genes induced was different for bulk and rhizosphere soils. Together, these results provide an unprecedented view of microbial gene expression in the plant rhizosphere during phytoremediation.     

Metal accumulation and tolerance in wetland plants

This paper briefly reviews the progress in studies of wetland plants in terms of heavy metal pollution. The current research mainly includes the following areas: (1) metal uptake, translocation, and distributions in wetland plants and toxicological effects on wetland plants, (2) radial oxygen loss (ROL) of wetland plants and its effects on metal mobility in rhizosphere soils, (3) constitutional metal tolerance in wetland plants, and (4) mechanisms of metal tolerance by wetland plants. Although a number of accomplishments have been achieved, many issues still remain unanswered. The future research effort is likely to focus on the ROL of wetland plants affecting metal speciation and bioavailability in rhizosphere soils, and the development of rhizosphere management technologies to facilitate and improve practical applications of phytoremediation of metalpolluted soils.     

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.     

Metal accumulation and tolerance in wetland plants

This paper briefly reviews the progress in studies of wetland plants in terms of heavy metal pollution. The current research mainly includes the following areas: (1) metal uptake, translocation, and distributions in wetland plants and toxicological effects on wetland plants, (2) radial oxygen loss (ROL) of wetland plants and its effects on metal mobility in rhizosphere soils, (3) constitutional metal tolerance in wetland plants, and (4) mechanisms of metal tolerance by wetland plants. Although a number of accomplishments have been achieved, many issues still remain unanswered. The future research effort is likely to focus on the ROL of wetland plants affecting metal speciation and bioavailability in rhizosphere soils, and the development of rhizosphere management technologies to facilitate and improve practical applications of phytoremediation of metal-polluted soils.     

增施CO2对植物修复土壤DEHP污染的影响

修复效率低一直是植物修复技术需要解决的关键问题之一.基于我国的CO₂减排压力和CO₂对植物生长的必要性,选择C3植物绿豆和C4植物玉米作为修复植物,以DEHP为目标污染物,探索增施CO₂对植物修复土壤DEHP污染的影响.结果表明: DEHP对两种植物生长和根际微环境都产生了抑制性影响.增施CO₂后,两种植物地上干质量显著增加,叶片SOD酶活性明显下降,根际土壤碱性磷酸酶活性增加,根际微生物群落结构改变,根际耐DEHP胁迫微生物数量增加,表明增施CO₂对促进植物生长、增强植物抗DEHP胁迫能力、改善根际微环境有积极作用.增施CO₂还促进了两种植物对DEHP的吸收,特别是植物地下部分.这些共同作用导致增施CO₂后的两种植物根际DEHP残留浓度明显下降,土壤污染植物修复效率提高.整体上看,增施CO₂对C3植物绿豆的影响明显大于C4植物玉米.可以将增施CO₂ 作为强化植物修复过程的措施之一.     

Influence of the zinc hyperaccumulator Thlaspi caerulescens J. & C. Presl. and the nonmetal accumulator Trifolium pratense L. on soil microbial populations

Metal hyperaccumulator plants like Thlaspi caerulescens J. & C. Presl. are used for phytoremediation of contaminated soils. Since little is known about the rhizosphere of hyperaccumulators, the influence of T. caerulescens was compared with the effects of Trifolium pratense L. on soil microbes. High- and low-metal soils were collected near a zinc smelter in Palmerton, Penn. Soil pH was adjusted to 5.8 and 6.8 by the addition of Ca(OH)2. Liming increased bacterial populations and decreased metal toxicity to levels allowing growth of both plants. The effects of the plants on total (culturable) bacteria, total fungi, as well as cadmium- and zinc-resistant populations were assessed in nonrhizosphere and rhizosphere soil. Both plants increased microbial populations in rhizosphere soil compared with nonrhizosphere soil. Microbial populations were higher in soils planted with T. pratense, but higher ratios of metal-resistant bacteria were found in the presence of T. caerulescens. We hypothesize that T. caerutescens acidifies its rhizosphere. Soil acidification in the rhizosphere of T. caerulescens would affect metal uptake by increasing available metals around the roots and consequently, increase the selection for metal-resistant bacteria. Soil acidification may be part of the hyperaccumulation process enhancing metal uptake from soil.     

Rhizosphere Microbial Characterization in Petroleum-Contaminated Soil

Contamination of soil with petroleum compounds is of concern worldwide. Although there are a variety of physical and chemical technologies available to remediate petroleum waste sites, biological methods are often used due to lower cost and public acceptance. Growth and enhanced activity of microbial communities in contaminated soil is a key factor for the success of bioremediation. Establishing vegetation in petroleum-contaminated soil may enhance microbial activity and remediation success even further by providing root exudates to the rhizosphere microorganisms. In this study, microorganisms were characterized in petroleum-contaminated soils and sediments quantitatively and qualitatively based on enumeration and metabolic diversity assessments. Contaminated soils and sediments were obtained from a phytoremediation field demonstration project in California. Microbial numbers in the unvegetated soil, based on plate counts and most probable number of hydrocarbon degraders, were significantly lower than the vegetated soils. Metabolic microbial characterization using BIOLOG was also conducted and based on principle component analysis (PCA), there was a distinct difference between the metabolic diversity of microbial communities in vegetated and unvegetated soils. Results from this research indicate that the presence and type of plants, and level of contamination may greatly influence microbial communities in polluted soils.     

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.     

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.     

Phytoremediation: novel approaches to cleaning up polluted soils

Environmental pollution with metals and xenobiotics is a global problem, and the development of phytoremediation technologies for the plant-based clean-up of contaminated soils is therefore of significant interest. Phytoremediation technologies are currently available for only a small subset of pollution problems, such as arsenic. Arsenic removal employs naturally selected hyperaccumulator ferns, which accumulate very high concentrations of arsenic specifically in above-ground tissues. Elegant two-gene transgenic approaches have been designed for the development of mercury or arsenic phytoremediation technologies. In a plant that naturally hyperaccumulates zinc in leaves, approximately ten key metal homeostasis genes are expressed at very high levels. This outlines the extent of change in gene activities needed in the engineering of transgenic plants for soil clean-up. Further analysis and discovery of genes for phytoremediation will benefit from the recent development of segregating populations for a genetic analysis of naturally selected metal hyperaccumulation in plants, and from comprehensive ionomics data--multi-element concentration profiles from a large number of Arabidopsis mutants.     

Screening of As-Accumulating Plants Using a Foliar Application and A Native Accumulation of As

The discovery of novel accumulating plants is useful for efficient phytoremediation due to the demands of various conditions of impacted sites such as land use, soil properties, concentration of pollutants, and climate. In the present study, we investigated foliar application or a field with highly bioavailable arsenic (As) to screen As-accumulating plants. Plants grown in the downstream of a hot springs area were analyzed for native As accumulation and As foliar application, and the rhizosphere soils were collected. The water-soluble As in the rhizosphere soils had a high average, 144 μg/kg, whereas total As was similar to normal soil in Japan. Among 34 herbaceous plants and 17 woody plants, Chelidonium majus var. asiaticum accumulated a relatively high As level, 8.07 mg/kg DW (93.6% of As added), that was not revealed by native accumulation. In a further pot experiment, C. majus accumulated a moderately high As level (314 mg/kg DW) in the roots but not in the shoot (30.1 mg/kg DW), and exhibited a low transfer factor (TF = 0.096). Thus, a foliar application would be a simple and high-throughput method to screen plants that accumulate and tolerate As. C. majus would be useful as a tool for phytostabilization of As.     

Phenotypic characterization of microbes in the rhizosphere of Alyssum murale

Metal hyperaccumulator plants like Alyssum murale are used for phytoremediation of Ni contaminated soils. Soil microorganisms are known to play an important role in nutrient acquisition for plants, however, little is known about the rhizosphere microorganisms of hyperaccumulators. Fresh and dry weight, and Ni and Fe concentrations in plant shoots were higher when A. murale was grown in non-sterilized compared to sterilized soils. The analysis of microbial populations in the rhizosphere of A. murale and in bulk soils demonstrated that microbial numbers were affected by the presence of the plant. Significantly higher numbers of culturable actinomycetes, bacteria and fungi were found in the rhizosphere compared to bulk soil. A higher percent of Ni-resistant bacteria were also found in the rhizosphere compared to bulk soil. Percentage of acid producing bacteria was higher among the rhizosphere isolates compared to isolates from bulk soil. However, proportions of siderophore producing and phosphate solubilizing bacteria were not affected by the presence of the plant. We hypothesize that microbes in the rhizosphere of A. murale were capable of reducing soil pH leading to an increase in metal uptake by this hyperaccumulator.     

Heavy metal pollution in aquatic ecosystems and its phytoremediation using wetland plants: an ecosustainable approach

This review addresses the global problem of heavymetal pollution originating from increased industrialization and urbanization and its amelioration by using wetland plants both in a microcosm as well as natural/field condition. Heavymetal contamination in aquatic ecosystems due to discharge of industrial effluents may pose a serious threat to human health. Alkaline precipitation, ion exchange columns, electrochemical removal, filtration, and membrane technologies are the currently available technologies for heavy metal removal. These conventional technologies are not economical and may produce adverse impacts on aquatic ecosystems. Phytoremediation of metals is a cost-effective "green" technology based on the use of specially selected metal-accumulating plants to remove toxic metals from soils and water. Wetland plants are important tools for heavy metal removal. The Ramsar convention, one of the earlier modern global conservation treaties, was adopted at Ramsar, Iran, in 1971 and became effective in 1975. This convention emphasized the wise use of wetlands and their resources. This review mentions salient features of wetland ecosystems, their vegetation component, and the pros and cons involved in heavy metal removal. Wetland plants are preferred over other bio-agents due to their low cost, frequent abundance in aquatic ecosystems, and easy handling. The extensive rhizosphere of wetland plants provides an enriched culture zone for the microbes involved in degradation. The wetland sediment zone provides reducing conditions that are conducive to the metal removal pathway. Constructed wetlands proved to be effective for the abatement of heavymetal pollution from acid mine drainage; landfill leachate; thermal power; and municipal, agricultural, refinery, and chlor-alkali effluent. the physicochemical properties of wetlands provide many positive attributes for remediating heavy metals. Typha, Phragmites, Eichhornia, Azolla, Lemna, and other aquatic macrophytes are some of the potent wetland plants for heavy metal removal. Biomass disposal problem and seasonal growth of aquatic macrophytes are some limitations in the transfer of phytoremediation technology from the laboratory to the field. However, the disposed biomass of macrophytes may be used for various fruitful applications. An ecosustainable model has been developed through the author's various works, which may ameliorate some of the limitations. The creation of more areas for phytoremediation may also aid in wetlands conservation. Genetic engineering and biodiversity prospecting of endangered wetland plants are important future prospects in this regard.     

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.     

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.     

Phytoremediation of Organic Contaminants: A Review of Phytoremediation Research at the University of Washington

As overwhelmingly positive results have become available regarding the ability of plants to degrade compounds such as trichloroethylene, phytoremediation studies are expanding. Studies to determine the potential for phytoremediation of fully chlorinated compounds, such as carbon tetrachloride and tetrachloroethylene, brominated compounds, such as ethylene dibromide and dibromochloropropane, and nonhalogenated compounds, such as methyl-t-butyl ether (MTBE), are underway. When using phytoremediation, it is important to select not only a plant that is capable of degrading the pollutant in question, but also one that will grow well in that specific environment. In ecologically sensitive areas, such as the Hawaiian Islands, only plants native to the area can be used. One way to supplement the arsenal of plants available for remedial actions is to utilize genetic engineering tools to insert into plants those genes that will enable the plant to metabolize a particular pollutant. Hybrid technologies, such as using plants in pumping and irrigation systems, also enable plants to be used as a remedial method when the source of the pollutant is beyond the reach of plant roots, or when planting space directly over the pollutant is unavailable or restricted. Thus, the potential uses of phytoremediation are expanding as the technology continues to offer new, low-cost remediation options.     

Copper concentration in plants and in the rhizosphere as influenced by the iron status of tomato (Lycopersicon esculentum L.)

Changes of metal concentration that occur in the rhizosphere may arise from several processes including variation in the concentration of complexing ligands, pH or redox potential that can be influenced by the Fe status of the plant. The aim of this study was to assess for both acidic and calcareous, Cu-contaminated soils how Cu concentration in plants and in the rhizosphere was affected by the Fe status of a strategy I plant species. The change of soil solution pH, total solution Cu concentration and soil redox potential was monitored for 8 days in the rhizosphere of tomato (Lycopersicon esculentum L.) in response to contrasting Fe supply. The concentration of Cu in roots was enhanced under Fe deficiency in the acidic soils. Shoot Cu however did not vary with the Fe status of the plant. The plant Fe status had little effect on rhizosphere pH, redox potential or Cu concentration in solution in either acidic or calcareous soils. Marked differences in pH and solution Cu concentration were observed between rhizosphere and uncropped soils. Roots induced an increase in pH of acidic soils and a decrease in solution Cu concentration in all soils. The decrease in solution Cu concentration in acidic soils may be explained by the increase in rhizosphere pH. The proposed device provided new data on the fate of Cu in the rhizosphere and showed a positive correlation for the four soils considered together between the total Cu concentration in soil solution and root Cu concentration.