Biotechnology Advances 28 (2010) 367374

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Biotechnology Advances
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Research review paper

Using soil bacteria to facilitate phytoremediation

Bernard R. Glick
Department of Biology, University of Waterloo, 200 University Avenue West, Waterloo, Ontario, Canada N2L 3G1

a r t i c l e

i n f o

a b s t r a c t
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 eld. As an adjunct to various phytoremediation strategies and as part of an effort to make this technology more efcacious, 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 eld studies might be facilitated. 2010 Elsevier Inc. All rights reserved.

Article history: Received 23 September 2009 Received in revised form 2 February 2010 Accepted 2 February 2010 Available online 8 February 2010 Keywords: Phytoremediation Organics Metals Environmental contamination Soil bacteria Plant growth-promoting bacteria

Contents Introduction . . . . . . . . . . . . . . . . . . . Phytoremediation . . . . . . . . . . . . . . . . Phytoremediation of organics facilitated by bacteria 3.1. Biodegradative bacteria . . . . . . . . . . 3.2. Plant growth-promoting bacteria . . . . . . 3.3. An optimal system? . . . . . . . . . . . . 4. Phytoremediation of metals facilitated by bacteria . 5. Future prospects . . . . . . . . . . . . . . . . . Acknowledgements . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . 1. 2. 3. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 367 368 368 368 369 370 370 370 372 372

1. Introduction The population of planet Earth will reach the 7 billion mark around the beginning of 2010 and, at the current rate of increase, it is estimated that it will be around 8 billion by 2019 (http://www.ibiblio.org/lunarbin/ worldpop). Moreover, around 40% of deaths worldwide are caused by water, air and soil pollution and such environmental degradation, coupled with the growth in world population, are (considered to be) major causes behind the rapid (global) increase in human disease (http://www.sciencedaily.com/releases/2007/08/070813162438.htm). Thus, as a consequence of both increasing population and industrial

Tel.: + 1 519 888 4567x32058; fax: + 1 519 746 0614. E-mail address: glick@sciborg.uwaterloo.ca. 0734-9750/$ see front matter 2010 Elsevier Inc. All rights reserved. doi:10.1016/j.biotechadv.2010.02.001

technology, humanity has created a situation where many life forms, including humans, are increasingly at risk. That is, until relatively recently, it was generally believed that earth's atmospheric, terrestrial, and aquatic systems were sufcient to absorb and break down wastes from population centers, industry, and agriculture. Unfortunately, we now know that this is not true. Thus, notwithstanding recent global efforts to curb human activities that are detrimental to the environment and human health, we are faced with a world that is highly contaminated with a range of toxic metals and organic compounds. The problem of toxic waste disposal is enormous. For example, it was estimated that in 1993 approximately 275 million tons of hazardous waste was produced in the United States (Ziegler, 1993). Moreover, in 1996, in the United States the Environmental Protection Agency listed 39,925 sites on its inventory of uncontrolled waste sites (DeRosa et al., 1996). Of the many chemicals found in hazardous waste sites in the

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United States, the Agency for Toxic Substances and Disease has created a list of the 275 hazardous substances that pose the greatest threat to human health (http://www.atsdr.cdc.gov/cercla/07list.html). The top ten priority substances are given in Table 1. To avoid the toxicity associated with these hazardous compounds, several technologies and methods have been developed to remove them from polluted soils. Many of these methods include the physical removal of soil to landll sites or extraction through chemical or physical means. These techniques are rapid but, unfortunately, they are costly from both an economic and an environmental point of view, and could potentially have a deleterious impact on soil physical, chemical, and biological properties. In addition, the removal from the environment of many toxic compounds is complicated by the numerous classes and types of these chemicals. For example, many soils are contaminated with one or more metals, other inorganic compounds, radioactive material or various organic compounds. Of these, the metals may include lead, zinc, cadmium, selenium, chromium, cobalt, copper, nickel or mercury; the other inorganic compounds might include arsenic, sodium, nitrate, ammonia or phosphate; the radioactive compounds may be uranium, cesium or strontium; and the organic compounds may include chlorinated solvents like trichloroethylene; explosives such as trinitrotoluene (TNT) and 1,3,5trinitro-1,3,5-hexahydrotriazine (RDX); petroleum hydrocarbons including benzene, toluene and xylene (BTX), polycyclic aromatic hydrocarbons (PAHs); and pesticides such as atrazine and bentazon.

2. Phytoremediation While some organic compounds can be metabolized (i.e., remediated) by bacteria that may either be found in or added to the soil, in the absence of plants, this process is usually slow and inefcient, in part as a consequence of the relatively low numbers of these degradative microorganisms in soil (Brookes and McGrath, 1984). On the other hand, the use of plants to remediate polluted soils (i.e., phytoremediation; Salt et al., 1995) is a potentially clean, effective and relative inexpensive technology that is likely to be readily accepted by a concerned public. However, this approach also has its drawbacks, as few plant species can naturally tolerate and/or accumulate high concentrations of the above mentioned environmental contaminants. Some organic compounds can be directly degraded and completely mineralized by plant enzymes through phytodegradation (Alkorta and Garbisu, 2001; Wild et al., 2005); many plants produce, and often secrete to the environment, enzymes that can degrade a wide range of organic compounds. However, inorganic pollutants cannot be degraded. Inorganic pollutants must either be stabilized in the soil to make them less bioavailable (i.e., phytostabilization); extracted, transported, and accumulated in plant tissues (i.e., phytoextraction); or transformed into volatile forms (i.e., phytovolatilization) (Pilon-Smits, 2005). Phytoremediation efciency for metals is often limited by the bioavailability of the metal in soil, plant root development, and the level of tolerance of the plant to each particular metal (Pilon-Smits, 2005).

Unfortunately, many of the plants that are most effective at removing metals from the soil, i.e., hyperaccumulators such as Thlaspi caerulescens (Alpine pennycress) and Alyssum bertolonii, are small and slow growing, thus reducing their potential for metal phytoextraction from soil on a large scale (Khan et al., 2000). To be effective for the remediation of metal polluted soils, plants must be tolerant to one or more metals, highly competitive, fast growing, and produce a high aboveground biomass. Because of their high biomass and extensive root system, trees are considered to be attractive for phytoremediation; however, metal accumulation by trees is generally low. Metal phytoextraction (as well as plant growth) can sometimes be facilitated by soil microorganisms living in intimate association with plant roots (Shilev et al., 2001). In addition, the biodegradation of recalcitrant organic compounds in the soil is often enhanced around the roots of plants. Following root exudation, the proliferation of specic groups of microorganisms, able to aggressively colonize the root surface and affect plant growth, occurs (Kloepper et al., 1989). As a consequence of the high level of nutrients that plants release into the soil as root exudates, the concentration of bacteria in the immediate vicinity of plant roots (i.e. the rhizosphere) is typically 10- to 1000-fold greater than the bacterial concentration that is found in the bulk soil. Some rhizosphere microorganisms can act directly on organic pollutants using their own degradative capabilities (phytostimulation or rhizodegradation) (Kuiper et al., 2004). As well, some soil bacteria can positively affect plants by improving growth and health (Glick, 1995), enhancing root development (Gamalero et al., 2002, 2004), or increasing plant tolerance to various environmental stresses (Glick, 2004). In turn, larger and healthier plants are better able to phytoremediate both organic and inorganic contaminants. This review provides an overview of the involvement of soil bacteria in assisted phytoremediation. Since various aspects of the work described here have previously been reviewed, from several different perspectives (Glick, 2003; Khan, 2005; Newman and Reynolds, 2005; Pilon-Smits, 2005; Krmer, 2005; Pilon-Smits and Freeman, 2006; Arshad et al., 2007; Jing et al., 2007; Zhuang et al., 2007; Doty, 2008; Kamaludeen and Ramasamy, 2008; Reichenauer and Germida, 2008; Yang et al., 2009; Gamalero et al., 2009; Gerhardt et al., 2009; Rajkumar et al., 2009; Weyens et al., 2009), this article is directed toward developing an understanding of the underlying principles of bacterially assisted phytoremediation as elaborated in the more recent scientic literature. 3. Phytoremediation of organics facilitated by bacteria Bacteria may interact with and affect the growth of plants in a variety of ways. Some bacteria are phytopathogenic and actively inhibit plant growth; others (plant growth-promoting bacteria) can facilitate the growth of plants using a wide range of different mechanisms; and there are a large number of soil bacteria that do not appear to affect the growth of plants one way or the other, although this may vary as a function of a range of different soil conditions (Glick, 1995). 3.1. Biodegradative bacteria

Table 1 U.S. Department of Health and Human Services, Agency for Toxic Substances and Disease Registry for 2007 (http://www.atsdr.cdc.gov/cercla/07list.html). Rank 1 2 3 4 5 6 7 8 9 10 Substance Arsenic Lead Mercury Vinyl chloride Polychlorinated biphenyls Benzene Cadmium Polycyclic aromatic hydrocarbons Benzo(A)Pyrene Benzo(B)Fluoranthene

It has been known for some time that many soil bacteria are able to degrade toxic organic compounds (Chakrabarty, 1981). With the discovery of a number of soil microorganisms that are capable of degrading xenobiotic chemicals including herbicides, pesticides, refrigerants, solvents, and other organic compounds, the notion that microbial degradation might provide a reasonable and effective means of disposing of toxic chemical wastes gained credence. Pseudomonas spp. are the most predominant group of soil microorganisms that biodegrade complex organic compounds, a process that typically requires the concerted efforts of several different enzymes. The genes that code for the enzymes of these biodegradative pathways are often located on large ( 50 kb to 200 kb) plasmids (Ghosal et al., 1985; Cork and Krueger, 1991).

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Degradative bacteria enzymatically convert xenobiotic, nonhalogenated aromatic compounds to either catechol or protocatechuate, compounds that are readily metabolized by almost all organisms. Halogenated aromatic compounds, which are the main components of most pesticides and herbicides, may also be degraded by these plasmidencoded enzymes, however, their rate of degradation is much slower. While addition of biodegradative bacteria to contaminated soils (i.e. bioremediation) is commonly effective at facilitating the breakdown of the contaminants in a laboratory setting, in the environment successful bioremediation is often more difcult to achieve. Equally problematic is the addition of plants to contaminated soils (i.e. phytoremediation) to metabolize and thereby remove toxic compounds from the soil. That is because even with plants that are somewhat tolerant of soil contaminants, plant growth is generally signicantly reduced so that the plants are unable to attain sufcient biomass to allow them to efciently degrade the contaminant within a reasonable time frame. One strategy that overcomes some of the limitations of both conventional bioremediation and phytoremediation includes utilizing biodegradative bacteria together with plants. In the past 15 years or so there have a number of published studies where scientists have successfully used this approach to remove organic toxicants from the soil. All of these biodegradative bacteria bind efciently to plant roots and some of them colonize the interior of the plant as well (i.e. they are endophytic). In addition, some of these biodegradative bacteria have plant growth-promoting, as well as biodegradative, activity. While most of the reported studies have been performed under more controlled laboratory, growth chamber or greenhouse conditions, a few studies have included eld trials. To date, the combined use of plants and biodegradative bacteria has been exploited/applied to remove petroleum products (Escalante-Espinosa et al., 2005; Radwan et al., 2005; Alarcn et al., 2008; Lin et al., 2008; AlAwadhi et al., 2009), polycyclic and other aromatic hydrocarbons (Daane et al., 2001; Barac et al., 2004; Taghavi et al., 2005; Sheng and Gong, 2006; Sheng et al., 2008d; Sheng et al., 2009; Germaine et al., 2009) and a variety of halogenated compounds (Brazil et al., 1995; Crowley et al., 1996; Sicilano and Germida, 1997; Yee et al., 1998; Nakamura et al., 2004; Leigh et al., 2006; Germaine et al., 2006; Liu et al., 2007; Al-Qurainy and AbdelMegeed, 2009; Uhlik et al., 2009) from contaminated soils. In addition to degrading polycyclic aromatic hydrocarbons per se, it has been suggested that some bacteria may make these relatively insoluble compounds more bioavailable. This could occur through the formation of a bacterial biolm directly on the surface of some crystal-like polycyclic aromatic hydrocarbons. This mechanism is likely to exist for only a limited number of bacteria that contain hydrophobic external surfaces. Bacteria with more hydrophilic surfaces would be dependent upon the (limited) concentration of polycyclic aromatic hydrocarbons in the bulk liquid. 3.2. Plant growth-promoting bacteria Plant growth-promoting bacteria may facilitate plant growth either indirectly or directly (Glick, 1995). The ability of plant growth-promoting bacteria to act as biocontrol agents against phytopathogens and thus indirectly stimulate plant growth may result from any one of a variety of mechanisms including antibiotic production, depletion of iron from the rhizosphere, induced systemic resistance, production of fungal cell wall lysing enzymes, and competition for binding sites on the root (Glick, 1995; Glick et al., 2007a,b). There are several ways in which plant growthpromoting bacteria can directly facilitate plant growth. They may x atmospheric nitrogen and supply it to plantsusually a minor component of the benet that the bacterium provides to the plant; synthesize siderophores which can sequester iron from the soil and provide it to plant cells which can take up the bacterial siderophoreiron complex; synthesize phytohormones such as auxins, cytokinins and gibberelins, which can act to enhance various stages of plant growth; solubilize minerals such as phosphorus, making them more readily available for plant growth; and synthesize the enzyme 1-aminocyclopropane-1-

carboxylate (ACC) deaminase, which can lower plant ethylene levels (Glick, 1995; Glick et al., 2007a,b). A bacterium may directly affect plant growth and development using any one or more of these mechanisms. Since many plant growth-promoting bacteria possess several of these traits, a bacterium may utilize different traits at various times during the life cycle of the plant. Typically, plant growth-promoting bacteria have little or no measurable effect on plant growth when the plants are cultivated under optimal and stress free conditions. Traditionally, the mechanism most often invoked to explain the various direct effects of plant growth-promoting bacteria on plants is the production of phytohormones, and most of the attention has focused on the role of the phytohormone auxin (Brown, 1974; Patten and Glick, 1996; Garcia de Salamone et al., 2005). However, in the last few years it has been found that a number of plant growth-promoting bacteria contain the enzyme ACC deaminase (Kishore and Kishore, 1992; Jacobson et al., 1994; Glick et al., 1995; Burd et al., 1998; Kaneko et al., 2000; Belimov et al., 2001, 2005; Babalola et al., 2003; Ma et al., 2003; Ghosh et al., 2003; Dey et al., 2004; Mayak et al., 2004a; Hontzeas et al., 2005; Madhaiyan et al., 2006; Shaharoona et al., 2006; Blaha et al., 2006; Saravanakumar and Samiyappan, 2006; Nadeem et al., 2007; Yue et al., 2007; Rodriguez et al., 2008) and that this enzyme can cleave the plant ethylene precursor ACC, and thereby lower the level of the phytohormone ethylene in a developing or stressed plant. The plant hormone ethylene plays an important role in root initiation and elongation, nodulation, senescence, abscission and ripening as well as in stress signaling (Mattoo and Suttle, 1991; Abeles et al., 1992; Arshad and Frankenberger, 2002). As part of a stress response, it inhibits root elongation, nodulation and auxin transport, induces hypertrophies, speeds aging and promotes senescence and abscission. During periods of environmental stress, plants produce high levels of stress ethylene. Moreover, much of the growth inhibition that occurs as a consequence of an environmental stress is the result of the response of the plant to the increased levels of stress ethylene which exacerbates the response to the stressor. However, ACC deaminase-containing bacteria can lower plant ethylene levels in plants and thereby provide some protection against the inhibitory effects of various stresses. ACC deaminase-containing plant growth-promoting bacteria have been used to protect plants against growth inhibition caused by: ooding (Grichko and Glick, 2001; Farwell et al., 2007); the presence of organic toxicants (Glick, 2003; Huang et al., 2004, 2005; Reed and Glick, 2005; Gurska et al., 2009); the presence of a variety of different metals (Burd et al., 1998; Belimov et al., 2001, 2005; Nie et al., 2002; Glick, 2003; Reed and Glick, 2005; Dell'Amico et al., 2005; Safranova et al., 2006; Farwell et al., 2006); high salt (Mayak et al., 2004b; Saravanakumar and Samiyappan, 2006; Cheng et al., 2007); phytopathogens (Wang et al., 2000; Hao et al., 2007); and drought (Mayak et al., 2004a). One of the main effects of bacterial IAA is the enhancement of lateral and adventitious rooting leading to improved mineral and nutrient uptake and root exudation that in turn stimulates bacterial proliferation on the roots (Dobbelaere et al., 1999; Lambrecht et al., 2000; Steenhoudt and Vanderleyden, 2000). The role of IAA synthesized by the plant growthpromoting bacterium Pseudomonas putida GR12-2, which produces relatively low levels of the phytohormone, in the development of the canola roots was studied following construction of an IAA-decient mutant of this strain (Patten and Glick, 2002). Seed inoculation with wildtype GR12-2 induced the formation of tap roots that were signicantly longer than the roots from seeds treated with the IAA-decient mutant and the roots from uninoculated seeds. It has been suggested that plant growth-promoting bacteria synthesizing IAA may prevent the deleterious effects of environmental stresses (Lindberg et al., 1985; Frankenberger and Arshad, 1995). For example, IAA stimulated lengthening of the root and shoot of wheat seedling exposed to high levels of salt (Egamberdieva, 2009). An increased tolerance of Medicago truncatula against salt stress was also observed in plants nodulated by the IAA-overproducing strain Sinorhizobium meliloti DR-64 (Bianco and Defez, 2009); plants inoculated with this mutant

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accumulated a high amount of proline, and showed enhanced levels of the antioxidant enzymes superoxide dismutase, peroxidase, glutathione reductase, and ascorbate peroxidase compared with plants inoculated with the parental strain. In addition to the more well characterized effects of microbial auxin and ethylene on plant growth, a number of plant growth-promoting bacteria synthesize cytokinins which can stimulate the growth of different crops under both stressed and non-stressed conditions (Ortiz Castro et al., 2008). 3.3. An optimal system? Given the fact that there were a large number of variables in the experiments discussed in this section, it isn't possible to distill this information and come up with one ideal set of conditions for all organic contaminant phytoremediation experiments. These variables include differences in: plant type, soil composition, endogenous bacteria, the nature, concentration and range of the contaminants, the temperature range, and the type (and physiological state) of added bacteria. This complexity notwithstanding, careful examination of the data suggest that certain considerations may facilitate the phytoremediation of organic environmental contaminants. These include: (i) inclusion of bacteria that are either engineered or selected to be able to both promote plant growth (e.g. through the provision of IAA) and degrade soil contaminants are generally superior to bacteria that can only promote plant growth or degrade soil contaminants [The major hurdle to the successful use of genetically engineered bacteria is political and regulatory, rather than scientic, in that these organisms are not currently considered to be acceptable for deliberate release into the environment.]; (ii) plant growth-promoting activity that is based on the activity of ACC deaminase appears to be universally effective in promoting plant growth under stressful conditions and is therefore a key determinant in facilitating phytoremediation; and (iii) endophytic bacteria appear to be more effective than bacteria that bind exclusively to the plant rhizosphere. 4. Phytoremediation of metals facilitated by bacteria The phytoremediation of metals that need to be removed from the environment is technically much more difcult than the phytoremediation of organic compounds that can be broken down in situ either in plants or in the soil. The major limitations of most metal phytoextraction processes are (i) the bioavailability of the target metal(s) and (ii) the ability of various plants to accumulate metals within their aboveground biomass (Raskin and Ensley, 2000). Scientists sometimes attempt to increase metal bioavailability through the addition of various chelating agents, a strategy that often works on a small scale in the laboratory but is much less effective in the eld. A large number of plants have been tested for their ability to take up high levels of metals and then translocate those metals from roots to leaves and shoots, however, many so-called hyperaccumulating plants do not produce sufcient biomass to make this process efcient in the eld (Raskin and Ensley, 2000). Although the use of soil bacteria (often plant growth-promoting bacteria) as adjuncts in metal phytoremediation can signicantly facilitate the growth of plants in the presence of high (and otherwise inhibitory) levels of metals, the bacteria typically do little or nothing to increase metal bioavailability. Following the initial demonstration that a nickel resistant ACC deaminase-containing plant growth-promoting bacterium could decrease the toxicity of nickel to canola plants (Burd et al., 1998), there have been a large number of reports of facilitation of metal phytoextraction through the addition of plant growth-promoting bacteria (Table 2). In the reported studies, which encompass a range of different plants, metals, soils and bacteria, the effect of adding bacteria is typically to facilitate plant growth and, when lower and

less toxic levels of metals are present, to increase the amount of metal taken up by the plant. The bacteria that have been utilized for these studies are typically rst selected for resistance to the target toxic metal(s) and then they are often either selected or tested for the presence of the enzyme ACC deaminase and/or the ability to synthesize IAA and siderophores. These studies are done with the expectation that the selected metal resistant bacteria will be able to both proliferate and promote plant growth in the presence of high levels of toxic metals. While very few studies have denitively identied the precise mechanism(s) of plant growth promotion in the presence of metal, given that the totality of the data are consistent with the involvement of IAA, siderophores and ACC deaminase, most recent studies assume that demonstrating the presence of all or even some of these activities is sufcient to elaborate the mechanism. On the other hand, the data that most denitively demonstrates the role of IAA, ACC deaminase and siderophores in plant growth promotion utilizes bacterial mutants lacking or over-expressing these activities. This data suggests that (i) IAA promotes plant growth per se (Patten and Glick, 2002), (ii) ACC deaminase prevents stress ethylene from becoming overly inhibitory of plant growth (Glick et al., 2007b) and (iii) siderophores help the plants to acquire sufcient iron in the presence of overwhelming amounts of other (potentially competing) metals (Burd et al., 2000). In addition to the above mentioned mechanisms of plant growth promotion, several other bacterial traits may act to facilitate metal phytoremediation. For example, some researchers have genetically engineered the bacteria that they used as an adjunct to phytoremediation to produce various metal-binding peptides (Wu et al., 2006a; Ike et al., 2007) making some metals in the soil more bioavailable. In addition, a number of researchers have found that bacteria that facilitate phytoremediation sometimes have an active phosphate solubilization system which some workers believe may play a role in assisting metal uptake. As well, one study found that a bacterial strain that aided phytoremediation produced biosurfactants, again, possibly helping to make the metals more bioavailable (Sheng et al., 2008a). Finally, it is important to bear in mind that in nature approximately 90% of terrestrial plants are mycorrhizal, so that an understanding of the optimal conditions for phytoremediation in the eld must ultimately take the role of these benecial fungi into account.

5. Future prospects Bacterially assisted phytoremediation is a technology whose time has nearly arrived. In the past ten years, scientists have developed a much better understanding of precisely how various bacteria contribute to phytoremediation and the efcacy of these approaches has been demonstrated under laboratory conditions. For organic contaminants, this approach has already been found to be effective in the eld and it is likely that during the next ve to ten years its use will become widespread. That is not to say that no technical difculties remain, however, the problems appear to be tractable. On the other hand, to efciently phytoremediate metal-contaminated soils, it is rst necessary to address the problem of metal bioavailability. In this regard, it may be possible to genetically engineer bacteria to facilitate the bioavailability of metals that are tightly bound to soil particles. Finally, to employ bacterially assisted phytoremediation of either organic or metal contaminants on a large scale in the environment, it will be necessary to convince regulatory bodies in various jurisdictions that the deliberate release of selected, or even engineered, bacteria to the environment should be viewed not only as benign, but in fact as benecial. This will require scientists to ensure, based on a thorough understanding of the processes and interactions between plants, contaminants, soils and bacteria, that bacterially assisted phytoremediation is a highly reproducible and dependable process.

B.R. Glick / Biotechnology Advances 28 (2010) 367374 Table 2 Plant-bacteria combinations in various metal phytoremediation studies. Bacterium Kluyvera ascorbata SUD165 K. ascorbata SUD165, SUD165/26 Plant(s) Canola (Brassica napus) Canola, tomato (Lycopersicon esculentum), Indian mustard (Brassica juncea) Thlaspi caerulescens Metal(s) Nickel Nickel, lead, zinc Effect/Mechanism Increased biomass; ACC deaminase Increased biomass; ACC deaminase, siderophores

371

Reference Burd et al., 1998 Burd et al., 2000

Whiting et al., 2001 Enterobacter cloacae CAL2 Canola Arsenic Nie et al., 2002 Brevibacillus sp. Red clover (Trifolium Lead Decreased lead uptake; IAA Vivas et pratense) al., 2003 Microbacterium arabinogalactanolyticum Alyssum murale Nickel Increased nickel uptake; mechanism Abouunknown Shanab et al., 2003 Rhizosphere and endophytic bacteria Thlaspi goesingense Nickel ACC deaminase, siderophores Idris et al., 2004 Psuedomonas aspleni AC Canola Copper Increased biomass; IAA Reed and Glick, 2005 Variovorax paradoxus, Rhodoccus sp., Flavobacterium sp. Indian mustard Cadmium Increased root length; IAA, siderophores, Belimov et ACC deaminase al., 2005 P. putida KNP9 Mung bean (Vigna radiata) Cadmium, Increased biomass and decreased metal Tripathi et lead uptake; siderophores al., 2005 Rhizosphere bacteria Graminaceae grasses Cadmium, IAA, siderophores, ACC deaminase Dell'Amico zinc, nickel et al., 2005 Pseudomonas sp. A4, Bacillus sp. 32 Indian mustard Chromium Increased root and shoot length; IAA, Rajkumar siderophores, phosphate solubilization et al., 2006 Pseudomonas uorescens Sunower (Helianthus Arsenic Increased growth; mechanism unknown Shilev et annuus) al., 2006 P. uorescens, P. putida Canola Nickel Increased seed germination and growth; Ashour et mechanism unknown al., 2006 P. putida 06909 Sunower Cadmium Increased cadmium uptake and decreased Wu et al., 2006a toxicity; bacterium expresses a metalbinding peptide P. putida UW4, P. putida HS-2 Canola Nickel Increased biomass in the eld; IAA, ACC Farwell et deaminase al., 2006 Safranova P. brassicacearum AM3, P. marginalis Dp1 Pea (Pisum sativum) Cadmium Increased biomass and nutrient uptake; ACC deaminase et al., 2006 Sinorhizobium sp. Pb002 Indian mustard Lead Increased plant survival and lead uptake; Di ACC deaminase Gregorio et al., 2006 Increased biomass and metal Wu et al., Azotobacter chroococcum HKN-5 + B. Indian mustard Zinc, bioavailability 2006b megaterium HKP-1 + B. mucilaginosus HKK-1 copper, lead, cadmium B. subtilis SJ-101 Indian mustard Nickel Increased nickel uptake; IAA, phosphate Zaidi et al., solubilization 2006 Pseudomonas sp. RJ10, Bacillus sp. RJ16 Canola Cadmium Increased biomass and metal uptake; IAA Sheng and Xia, 2006 Nine different nickel resistant bacterial Alyssum murale Nickel Increased metal uptake; mechanism Aboustrains unknown Shanab et al., 2006 P. diminuta, Brevundimonas diminuta, Nitrobacteria irancium, Ochrobacterum Water hyacinth Chromium Increased metal uptake; mechanism Abouanthropi, B. cereus (Eichhornia crassipes) unknown Shanab et al., 2007 Mesorhizobium huakuii subsp. rengei B3 Chinese milk vetch Cadmium Increased metal accumulation; bacterium Ike et al., 2007 (Astragalus sinicus) expresses phytochelatin and metallothionein Li et al., Burkholderia cepacia Sedum alfredii Cadmium, Increased biomass, metal uptake and 2007 zinc translocation of metal to shoots; mechanism unknown P. putida ARB86 Arabidopsis thaliana Nickel Increased biomass and chlorophyll Someya et content al., 2007 Bradyrhizobium sp. RM8 Mung bean (also known as Nickel, Increased nodule number and plant Wani et greengram) zinc nutrition; IAA, siderophores al., 2007 Rhizobium sp. RP5 Pea Nickel, Increased nodule number and plant Wani et zinc nutrition; IAA, siderophores al., 2008 Burkholderia sp. J62 Indian mustard, corn (Zea Lead, Increased biomass and metal uptake; IAA, Jiang et al., mays), tomato cadmium siderophores, ACC deaminase 2008 Hussein, Increased metal uptake depending on Bacillus licheniformis, B. biosubtyl, B. Indian mustard Selenium, 2008 specic metalbacteria combination; thurnigiensis cadmium, chromium mechanism unknown (continued on next page)

Rhizosphere bacteria

Zinc

Increased zinc uptake; mechanism unknown Increased biomass; ACC deaminase

372 Table 2 (continued) Bacterium Enterobacter sp. NBRI K28

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Plant(s) Indian mustard

Metal(s) Nickel, zinc, chromium Nickel Cadmium

Effect/Mechanism Increased biomass and metal uptake; IAA, siderophores, ACC deaminase, phosphate solubilization Increased seed germination and biomass; siderophores, IAA, ACC deaminase Some increased biomass and cadmium uptake, IAA, siderophores, biosurfactant production Increased biomass and rooting, and decreased cadmium uptake; IAA, siderophores, ACC deaminase, phosphate solubilization Increased biomass; IAA, siderophores, ACC deaminase, phosphate solubilization

Reference Kumar et al., 2008 Rodriguez et al., 2008 Sheng et al., 2008a Ganesan, 2008

P. putida HS-2 Bacillus sp. J119

Canola Canola, corn, sudan grass (Sorghum vulgare var. sudanense), tomato Black gram (Vigna mungo)

P. aeruginosa MKRh3

Cadmium

Pseudomonas sp. 29C, Bacillus sp. 4C

Indian mustard

Nickel

Pseudomonas sp. M6, Pseudomonas jessenii M15

Castor bean (Ricinus communis)

Nickel, copper, zinc

Proteus vulgaris KNP3

Pigeon pea (Cajanus cajan) Copper

10 different rhizosphere bacteria

Goat willow (Salix caprea)

Cadmium, zinc Nickel

Pseudomonas sp.

Chickpea (Cicer arietinum)

P. uorescens G10, Microbacterium sp. G16

Canola

Lead

Bacillus edaphicus NBT Pseudomonas sp. RJ10, Bacillus sp. RJ16

Indian mustard Tomato

Lead Cadmium, lead Zinc Cadmium Copper

Flavobacterium sp. Streptomyces tendae F4 Achromobacter xylosoxidans Ax10

Orychophragmus violaceus Sunower Indian mustard

Enterobacter aerogenes, Rahnella aquatilis

Indian mustard

Nickel, chromium

Rajkumar and Freitas, 2008a Increased biomass; IAA, ACC deaminase, Rajkumar phosphate solubilization and Freitas, 2008b Increased germination, biomass and Rani et al., chlorophyll, and decreased metal uptake; 2008 mechanism unknown Kuffner et Increased metal uptake; IAA, al., 2008 siderophores, ACC deaminase all not involved Increased biomass and decreased metal Tank and uptake; siderophores Saraf, 2008 Increased biomass and metal uptake; IAA, Sheng et al., 2008b siderophores, ACC deaminase, both strains are endophytes Increased biomass; IAA, siderophores, Sheng et ACC deaminase al., 2008c He et al., Increased root length, above ground 2009 biomass and above ground metal; siderophores, IAA, ACC deaminase Increased root length, biomass, metal He et al., uptake; mechanism unknown 2010 Decreased metal uptake and increased Dimpka et iron content; siderophores al., 2009 Ma et al., Increased root and shoot length and 2009 biomass; ACC deaminase, phosphate solubilization, IAA Increased biomass and metal uptake; IAA, Kumar et siderophores, ACC deaminase, phosphate al., 2009 solubilization

Acknowledgements The work from our laboratory that is reviewed here was supported by funds from the Natural Sciences and Engineering Research Council of Canada. References
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