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Phytoremediation: Synergistic Use of Plants and Bacteria to Clean Up

the Environment

Article  in  Biotechnology Advances · September 2003

DOI: 10.1016/S0734-9750(03)00055-7 · Source: PubMed

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 Biotechnology Advances 21 (2003) 383 – 393
www.elsevier.com/locate/biotechadv

Phytoremediation: synergistic use of plants and

bacteria to clean up the environment
Bernard R. Glick *
Department of Biology, University of Waterloo, Waterloo, ON, Canada N2L 3G1

Abstract

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.
D 2003 Elsevier Inc. All rights reserved.

Keywords: Phytoremediation; Plants; Bacteria

1. The problem: environmental contamination

The removal from the environment of many potentially toxic compounds is complicated
by the numerous classes and types of these chemicals. For example, many soils are
contaminated with one or more metals, radioactive or inorganic compounds. Of these, the
metals may include lead, zinc, cadmium, selenium, chromium, cobalt, copper, nickel and
mercury; the radioactive compounds may be uranium, cesium or strontium; and the other

* Tel.: +1-519-888-4567; fax: +1-519-746-0614.

E-mail address: glick@sciborg.uwaterloo.ca (B.R. Glick).

0734-9750/03/$ - see front matter D 2003 Elsevier Inc. All rights reserved.
doi:10.1016/S0734-9750(03)00055-7
384 B.R. Glick / Biotechnology Advances 21 (2003) 383–393

inorganic compounds might include arsenic, sodium, nitrate, ammonia or phosphate. Soil
may become polluted with high concentrations of metals by either a natural phenomenon
such as proximity to an ore body, or as a consequence of industrial activities. The reme-
diation of heavily metal-contaminated soils often involves excavation and removal of soil to
‘‘secured’’ landfills, a technology that is expensive and requires site restoration. As an al-
ternative, in the past few years, several groups of scientists have begun to develop tech-
nological approaches to using certain plants to remove metal contaminants from the soil.
In addition to the above-mentioned inorganic compounds, soils and water systems may
also be contaminated with organic compounds including chlorinated solvents like
trichloroethylene; explosives such as trinitrotoluene (TNT) and 1,3,5-trinitro-1,3,5-hex-
ahydrotriazine (RDX); petroleum hydrocarbons including benzene, toluene and xylene
(BTX), polyaromatic hydrocarbons (PAHs); and pesticides such as atrazine and bentazon.
While many of these compounds can be metabolized by some soil bacteria, this process is
usually slow and inefficient, in part as a consequence of the relatively low numbers of
these degradative microorganisms in soil. However, there is mounting evidence that the
biodegradation of recalcitrant organic compounds in the soil is enhanced around the roots
of plants.

2. Phytoremediation: a solution to environmental contamination?

One recently developed method of environmental clean-up is called phytoremediation

(Cunningham and Berti, 1993; Kumar et al., 1995; Raskin et al., 1994, 1997). This
procedure may be defined as the use of plants to remove, destroy or sequester hazardous
substances from the environment. Phytoremediation of metals and other inorganic
compounds may take one of several forms: phytoextraction, the absorption and concen-
tration of metals from the soil into the roots and shoots of the plant; rhizofiltration, the use
of plant roots to remove metals from effluents; phytostabilization, the use of plants to
reduce the spread of metals in the environment; or phytovolatilization, the uptake and
release into the atmosphere of volatile materials such as mercury- or arsenic-containing
compounds. Phytoremediation of organic compounds may occur by phytostabilization;
phytostimulation, the stimulation of microbial biodegradation in the rhizosphere, the area
around the roots of plants; or by phytotransformation, the absorption and degradation of
organic contaminants by the plant.
Following the testing of a large number of different plants, a number of plants that can
naturally accumulate large amounts of metal have been identified and are being studied for
the phytoremediation of metals that are present in the environment. These plants, called
hyperaccumulators, are often found growing in areas with elevated metal concentrations in
the soil. Unfortunately, in the presence of very high concentrations of metals, even
hyperaccumulating plants attain only a small size. That is, high concentrations of metals
are inhibitory to the growth of plants, even those plants that are capable of hyper-
accumulating metals. Depending upon the amount of metal at a particular site and the type
of soil at that site, it could take 15– 20 years to completely remove the metal from the soil
and thereby remediate the site, even with hyperaccumulating plants. This is a timeframe
that is usually considered to be too slow for practical application.
 B.R. Glick / Biotechnology Advances 21 (2003) 383–393 385

A number of different types of plants are effective at stimulating the degradation of

organic molecules in the rhizosphere. Typically, these plants all have extensive and fibrous
roots, which form an extended rhizosphere; these plants include many common grasses as
well as corn, wheat, soybean, peas and beans. In addition, several varieties of trees can
take up and degrade some organic contaminants. For example, plants with phytotransfor-
mation activity may contain nitroreductases, which are useful for degrading TNT and other
nitroaromatics, dehalogenases for the degradation of chlorinated solvents and pesticides,
and laccases that can degrade anilines such as triaminotoluene.

3. Plant growth-promoting bacteria

Beneficial free-living soil bacteria are generally referred to as plant growth-promoting

rhizobacteria and are found in association with the roots of many different plants (Glick et
al., 1999). The high concentration of bacteria around the roots, i.e., in the rhizosphere,
presumably occurs because of the presence of high levels of nutrients (especially small
molecules such as amino acids, sugars and organic acids) that are exuded from the roots of
most plants, and can then be used to support bacterial growth and metabolism (Whipp,
1990; Bayliss et al., 1997; Penrose and Glick, 2001).
Plant growth-promoting bacteria can positively influence plant growth and develop-
ment in two different ways: indirectly or directly (Glick et al., 1999). The indirect
promotion of plant growth occurs when these bacteria decrease or prevent some of the
deleterious effects of a phytopathogenic organism. Bacteria can directly promote plant
growth by providing the plant with a compound that is synthesized by the bacterium or by
facilitating the uptake of nutrients from the environment by the plant. Plant growth-
promoting bacteria may: fix atmospheric nitrogen and supply it to plants; synthesize
siderophores which can solubilize and sequester iron from the soil and provide it to plant
cells; synthesize several different phytohormones including auxins and cytokinins which
can enhance various stages of plant growth; have mechanisms for the solubilization of
minerals such as phosphorus which then become more readily available for plant growth;
and contain enzymes that can modulate plant growth and development (Brown, 1974;
Davison, 1988; Kloepper et al., 1989; Lambert and Joos, 1989; Patten and Glick, 1996;
Glick et al., 1998). A particular bacterium may affect plant growth and development using
any one, or more, of these mechanisms and a bacterium may utilize different mechanisms
under different conditions. For example, bacterial siderophore synthesis is likely to be
induced only in soils that do not contain sufficient levels of iron. Similarly, bacteria do not
fix nitrogen when sufficient fixed nitrogen is available.

3.1. The role of ACC deaminase in the reduction of plant ethylene

In higher plants, ethylene is produced from L-methionine via the intermediates, S-

adenosyl-L-methionine (SAM) and 1-aminocyclopropane-1-carboxylic acid (ACC; Yang
and Hoffman, 1984). The enzymes involved in this metabolic sequence are SAM
synthetase, which catalyzes the conversion of methionine to SAM (Giovanelli et al.,
1980); ACC synthase, which is responsible for the hydrolysis of SAM to ACC and 5V-
386 B.R. Glick / Biotechnology Advances 21 (2003) 383–393

methylthioadenosine (Kende, 1989); and ACC oxidase, which metabolizes ACC to

ethylene, carbon dioxide and cyanide (John, 1991).
In 1978, an enzyme capable of degrading ACC was isolated from Pseudomonas sp.
strain ACP (Honma and Shimomura, 1978). Since then, ACC deaminase has been detected
in the fungus, Penicillium citrinum (Honma, 1993), and in a number of bacterial strains
(Klee and Kishore, 1992; Jacobson et al., 1994; Glick et al., 1995; Campbell and
Thomson, 1996; Burd et al., 1998; Belimov et al., 2001; Ghosh et al., 2003; Ma et al.,
2003) and in the yeast, Hansenula saturnus (Minami et al., 1998), all of which originated
from the soil either as soil sample isolates or as microbes typically found in the soil. This
enzyme cleaves the plant ethylene precursor, ACC, to produce ammonia and a-ketobu-
tyrate. Glick et al. (1998) proposed that microorganisms that contain the enzyme ACC
deaminase can all act to promote plant growth since they can act as a sink for ACC and
thereby lower ethylene levels in a developing or stressed plant.
Ethylene is important for normal plant development in plants as well as for their
response to stress (Deikman, 1997). Ethylene is important during the early phase of plant
growth; it is required by many plant species for seed germination, and the rate of ethylene
production increases during germination and seedling growth (Abeles et al., 1992).
However, high levels of ethylene lead to inhibition of root elongation. A model was
developed whereby plant growth-promoting bacteria that possess the enzyme ACC
deaminase and are bound to seeds or roots of seedlings, can reduce the amount of plant
ethylene and the extent of its inhibition on root elongation (Glick et al., 1998).
In this model, the plant growth-promoting bacteria bind to the surface of either the seed
or root of a developing plant; in response to tryptophan and other small molecules in the
seed or root exudates (Whipp, 1990; Bayliss et al., 1997; Penrose and Glick, 2001), the
plant growth-promoting bacteria synthesize and secrete indole acetic acid (IAA) (Patten
and Glick, 1996, 2002), some of which may be taken up by the plant. This IAA, together
with endogenous plant IAA, can either stimulate plant growth or induce the synthesis of
ACC synthase, which converts SAM to ACC.
A portion of the ACC produced by this latter reaction is exuded from seeds or plant
roots along with other small molecules normally present in seed or root exudates (Bayliss
et al., 1997; Penrose and Glick, 2001). The ACC in the exudates is taken up by the bacteria
and is subsequently converted by ACC deaminase to ammonia and a-ketobutyrate. This
decreases the amount of ACC outside the plant so that the plant must exude increasing
amounts of ACC in order to maintain the equilibrium between internal and external ACC
levels. Thus, in the first instance, the bacteria induce the plant to synthesize more ACC
than it would otherwise need and, secondly, it stimulates the exudation of ACC from the
plant. As a result of the ACC deaminase-containing bacteria lowering the ACC level
within the plant and acting as a sink for ACC, the amount of ethylene that is produced by
the plant is also reduced.
When tested, strains of ACC deaminase-containing plant growth-promoting bacteria
were found to reduce the amount of ACC that was detectable by HPLC, and hence the
ethylene levels in canola seedlings were also lowered (Penrose et al., 2001). Moreover,
ACC deaminase-containing plant growth-promoting bacteria promote root elongation in a
variety of (ethylene sensitive) plants (Hall et al., 1996). There is also a considerable
decrease in the levels of ‘‘stress ethylene’’—the accelerated biosynthesis of ethylene
 B.R. Glick / Biotechnology Advances 21 (2003) 383–393 387

associated with biological and environmental stresses, and pathogen attack (Morgan and
Drew, 1997)—in plants grown in the presence of ACC deaminase-containing plant
growth-promoting bacteria. The deleterious effects of flooding on tomato plants (Grichko
and Glick, 2001) were decreased and the shelf life of the petals of ethylene sensitive cut
flowers was prolonged following treatment with ACC deaminase-containing plant growth-
promoting bacteria (Nayani et al., 1998). Moreover, biocontrol strains of bacteria carrying
ACC deaminase genes were able to more effectively protect plants against various
phytopathogens: cucumber plants were protected against Pythium damping-off, and potato
slices, exposed to Erwinia caratovora in a small sealed bag, were protected against
Erwinia soft rot (Wang et al., 2000). In addition, canola seedlings grown in the presence of
high levels of nickel, produced much less ethylene when the seeds were inoculated with an
ACC deaminase-containing nickel-resistant plant growth-promoting strain that (Burd et
al., 1998). It appears that the ‘‘stress ethylene’’ produced in each of these situations, and
the damage caused by it, was reduced by the activity of ACC deaminase that lowered the
level of ethylene produced by the plant.

4. The use of plant growth-promoting bacteria in phytoremediation

4.1. Metals

While plants grown on metal contaminated soils might be able to withstand some of the
inhibitory effects of high concentrations of metals within a plant, two features of most plants
could result in a decrease in plant growth and viability. That is, in the presence of high levels
of metals, most plants (i) synthesize stress ethylene and (ii) become severely depleted in the
amount of iron that they contained. Moreover, plant growth-promoting bacteria may be used
to relieve some of the toxicity of metals to plants. This could occur in two different ways. As
indicated earlier, the use of ACC deaminase-containing plant growth-promoting bacteria
would be expected to decrease the level of stress ethylene in a plant growing in soil that
contained high levels of metal. In addition, plants are able to take up and utilize complexes
between bacterial siderophores and iron. Plant siderophores bind to iron with a much lower
affinity than bacterial siderophores so that in metal contaminated soils a plant is unable to
accumulate a sufficient amount of iron unless bacterial siderophores are present.
Our primary objective was the development of a phytoremediation system that could be
used to help to remove nickel from soil as inexpensively and as quickly as possible. Prior
to this work, there were reports in the scientific literature that indicated that Brassica
juncea (Indian mustard) was a nickel-hyperaccumulating plant and could be used for this
purpose. However, preliminary laboratory experiments indicated that the growth of Indian
mustard, and the related plant Brassica campestris (canola), which could also accumulate
high levels of nickel and other metals, was significantly inhibited by the presence of
moderate amounts of nickel in the soil. In an effort to overcome the inhibition of plant
growth by nickel, a bacterium was isolated from a nickel contaminated soil sample; the
bacterium was (i) nickel-resistant, (ii) able to grow at the cold temperatures (i.e., 5– 10 jC)
that one expects to find in nickel contaminated soil environments in Canada and (iii) an
active producer of ACC deaminase (Burd et al., 1998). In total, there were approximately
388 B.R. Glick / Biotechnology Advances 21 (2003) 383–393

4  103 nickel-resistant bacteria per gram dry weight of soil, or about 1% of the total
bacterial population that was culturable on Tris-buffered low-phosphate medium. In order
to isolate plant growth promoting bacteria, all of the nickel-resistant isolates were tested
for the ability to grow on minimal medium with ACC as the sole source of nitrogen (Glick
et al., 1995). Approximately 7% of the nickel tolerant strains also had the ACC+
phenotype. Finally, nickel-resistant bacterial strains that were also able to grow on ACC
were tested for the ability to produce siderophores. Based on the idea that bacterial
siderophores might facilitate the uptake of iron by plants, the best siderophore producing
strain (designated SUD165) was selected for subsequent study. This strain was charac-
terized by fatty acid analysis as Kluyvera ascorbata. In laboratory tests, it was ascertained
that K. ascorbata SUD165 not only had the above-mentioned traits but could also promote
plant growth in the presence of high levels of nickel (Burd et al., 1998; Ma et al., 2001). At
all levels of nickel tested (i.e., 1 – 6 mM Ni2 +), using both a low and a high level of
bacterial cell treatment (cell suspension absorbance at 600 nm of 0.025 or 0.50), with both
canola and tomato plants, with both roots and shoots, and both in gnotobiotic growth
pouches and in pots with soil, the addition of K. ascorbata SUD165 significantly
decreased the toxicity of the added nickel. Moreover, the protective effect of K. ascorbata
SUD165 increased as the density of the cell suspension increased. While the bacterium K.
ascorbata SUD165 did not change the amount of nickel taken up per milligram dry weight
of either roots or shoots and thus had no influence on amount of nickel accumulated by the
plant, it did lower the amount of ethylene that was evolved by plants treated with nickel in
comparison to treat with nickel in the absence of the bacterium. The simplest explanation
of the data is that the bacterium protects the plant against the inhibitory effects of nickel-
induced stress ethylene formation.
To improve the performance of K. ascorbata SUD165, the bacterium was grown on a
minimal medium that did not contain any measurable iron. Of the tens of thousands of
bacteria plated, only a few colonies were able to grow under these conditions. It was
reasoned that these bacteria probably contained a spontaneous mutation that caused the
overproduction of the bacterial siderophores. This siderophore overproduction enabled the
bacterium to sequester a sufficient amount of iron, even though the iron was present at
extremely low levels, to permit these bacteria to grow. The amount of siderophores
produced by each spontaneous mutant was quantified and the strain that produced the
highest level of siderophores (about 100-fold more than the wild-type) was designated K.
ascorbata SUD165/26 and selected for additional study (Burd et al., 2000).
When the wild-type bacterium and the siderophore overproducing mutant were tested
in the laboratory, as expected both of them were observed to promote the growth of
tomato, canola and Indian mustard plants in the presence of inhibitory levels (generally 2
mM) of nickel, lead or zinc. In addition, the siderophore overproducing mutant decreased
the inhibitory effect of the added metal on plant growth significantly more than the wild-
type bacterium. Heavy metal contamination of soil is often associated with iron-deficiency
in a range of different plant species (Mishra and Kar, 1974). The low iron content of plants
that are grown in the presence of high levels of heavy metals generally results in these
plants becoming chlorotic since iron deficiency inhibits both chloroplast development and
chlorophyll biosynthesis (Imsande, 1998). Moreover, iron deficiency causes the plant to
synthesize stress ethylene.
 B.R. Glick / Biotechnology Advances 21 (2003) 383–393 389

Once they have bound iron, microbial iron-siderophore complexes can be taken up by
plants and thereby serve as an iron source for plants (Bar-Ness et al., 1991). It was
therefore reasoned that the best way to prevent plants from becoming chlorotic in the
presence of high levels of heavy metals was to provide them with an associated
siderophore-producing bacterium that could provide a sufficient amount of iron to the
plant. Taken together, these results suggest a dual role for bacteria that facilitate plant
growth in the presence of heavy metals. On the one hand, the bacteria lower the level of
stress ethylene in the plant thereby allowing the plant to develop longer roots and thus
better establish itself during early stages of growth (Burd et al., 1998; Glick et al., 1998).
On the other hand, once the seedling is firmly established in the soil, the bacterium helps
the plant to acquire sufficient iron for optimal plant growth, in the presence of levels of
heavy metals that might otherwise make the acquisition of iron difficult (Burd et al., 2000).
When the siderophore overproducing mutant was tested in the field with soil that had
been contaminated with nickel over a period of many years, it was observed that both the
number of Indian mustard seeds that germinated in the nickel-contaminated soil, and the
size that the plants were able to attain was increased by 50 –100% by the addition of the
bacterium to the soil. While additional laboratory and field testing of the selected
bacterium is necessary, at this stage, the data look sufficiently promising to consider
commercialization.

4.2. Arsenate

Given the toxicity of arsenate to most plants, the development of a phytoremediation

scheme for the detoxification of arsenate-contaminated soils is not a simple matter. For
example, unlike what was observed with nickel, lead and zinc, arsenate-resistant plant
growth-promoting bacteria did not significantly protect plants from arsenate inhibition. On
the other hand, transgenic canola plants expressing a bacterial ACC deaminase under the
control of the cauliflower mosaic virus 35S promoter, which ensures that the transgene
will be expressed constitutively, were significantly more resistant to the toxic effects of
arsenate than were wild-type canola plants (Nie et al., 2002).
Following growth in the presence of arsenate, there were significant differences
between transgenic and non-transformed canola. Regardless of the presence or absence
of plant growth-promoting bacteria, a maximum of about 25% of the non-transformed
seeds germinated in the presence of arsenate. By contrast, about 70% of the transgenic
canola seeds germinated under the same conditions. Although a small ethylene pulse is
important in breaking seed dormancy in many plants, too much ethylene can inhibit plant
seed germination (Bewley and Black, 1985). In the presence of arsenate, ACC deaminase
may enhance the process of germination by hydrolyzing any excess ACC that forms as a
consequence of the stress, hence lowering the inhibitory level of ethylene in seeds.
The fresh and dry weights of canola roots and shoots, and the shoot chlorophyll
contents further support the hypothesis that lowering ethylene levels protects the plant
against arsenate inhibition. When grown in the presence of arsenate, the fresh and dry
weights of roots and shoots of transgenic canola, especially when they were treated with
the ACC deaminase-containing plant growth-promoting bacterium Enterobacter cloacae
CAL2, were much higher than with non-transformed canola. Other properties of this
390 B.R. Glick / Biotechnology Advances 21 (2003) 383–393

bacterial strain, in addition to ACC deaminase activity, may contribute to this result—the
bacterium synthesizes IAA, siderophores and antibiotics, all of which may stimulate plant
growth. In this regard, antibiotic-secreting plant growth-promoting bacterial strains can
inhibit the proliferation and subsequent invasion of phytopathogens, hence protecting
plants, already debilitated by arsenate in the soil, from further damage.
When biomass is considered in calculating the arsenate accumulation, each transgenic
canola plant accumulates approximately four times as much arsenate, on a dry-weight
basis, as non-transformed canola. The higher rate of germination of transgenic canola also
contributes to the total amount of arsenate accumulation. The significant increase of
arsenate accumulation made by transgenic canola in conjunction with plant growth-
promoting bacteria makes phytoremediation much more efficient. However, despite the
improved performance of transgenic versus wild-type canola, it is unlikely that this result
can be the basis of a large-scale process that is useful for the phytoremediation of arsenate
contaminated soils.

4.3. Organic contaminants

Polycyclic aromatic hydrocarbons are a particularly recalcitrant group of contaminants

and are known to be highly persistent in the environment. There are many sources for PAH
contamination in soils including creosote, fossil fuel processing and steel production. It is
expensive and time consuming to remediate persistent contaminants such as PAHs from
soils, and the techniques used to remediate PAH-contaminated soils tend to be inefficient.
Physical removal of PAH-contaminated soil and washing of those soils with solvents is
expensive, and has met with mixed results. Bioreactors have been attempted, however the
contaminated soils must still be brought to the reactor for the cleanup. This is expensive,
and it damages the natural structure and texture of the soil. In situ microbial remediation
(i.e., bioremediation) has been attempted, but it is difficult to generate sufficient biomass
in natural soils to achieve an acceptable rate of movement of hydrophobic PAHs (which
are often tightly bound to soil particles) to the microbes where they can be degraded. As
well, PAHs are very stable compounds, and the initial oxidation step is biologically slow
and metabolically expensive. Thus, in most situations, microbial remediation alone is
probably too slow to be a realistic approach to this problem. In addition, relatively few
microorganisms can use high molecular weight PAHs as a sole carbon source. More
recently, there have been some improvements in the strategies for bacterial remediation of
contaminated soil, including inoculation with bacteria that were selected from PAH
contaminated sites, or supplementing contaminated soils with nutrients (Suthersan,
2002). Nonetheless, there has only been limited success with these techniques. For
bioremediation to be effective, the overall rate of PAH removal and degradation must
be accelerated above current levels. One way to achieve this is to increase the amount of
biomass in the contaminated soil. For this reason, the use of higher plants (i.e.,
phytoremediation) has received considerable attention (Cunningham et al., 1995; Cun-
ningham and Ow, 1996).
The advantages of phytoremediation compared to other approaches are: (1) it preserves
the natural structure and texture of the soil; (2) energy is primarily derived from sunlight;
(3) high levels of biomass in the soil can be achieved; (4) it is low in cost; and (5) it has the
 B.R. Glick / Biotechnology Advances 21 (2003) 383–393 391

potential to be rapid. Although using plants for remediation of persistent contaminants

may have advantages over other methods, many limitations exist for the large-scale
application of this technology. For example, many plant species are sensitive to
contaminants including PAHs so that they grow slowly and it is time consuming to
establish sufficient biomass for meaningful soil remediation. In addition, in most
contaminated soils, the number of microorganisms is depressed so that there are not
enough bacteria either to facilitate contaminant degradation or to support plant growth. To
remedy this situation, both degradative and plant growth-promoting bacteria may be added
to the plant rhizosphere.
Phytoremediation (i.e., degradation of organics in the presence of plants) alone is not
significantly faster than bioremediation (i.e., where biodegradation of the organics is by
microorganisms independent of plants) for removal of PAHs that include three rings or less
(Huang, El-Alawi, Penrose, Glick and Greenberg, submitted for publication), although
phytoremediation outperformed bacterial treatment with respect to removal of the larger,
more strongly soil bound PAHs.
Cultivating plants together with plant growth-promoting bacteria allowed the plants to
germinate to a much greater extent, and then to grow well and rapidly accumulate a large
amount of biomass. In addition, the plant growth-promoting bacteria in these experiments
increased PAH removal; this is probably due to an alleviation of a portion of the stress
imposed upon the plant by the presence of the PAHs. The plant growth-promoting bacteria
increased seed germination and plant survival in heavily contaminated soils, decreased the
plant dry weight to fresh weight ratio, increased the plant water content, helped plants to
maintain their chlorophyll contents and chlorophyll a/b ratio, and promoted plant root
growth. As a consequence of the treatment of plants with plant growth-promoting bacteria,
the plants provide a greater sink for the contaminants since they are better able to survive
and proliferate.

5. Conclusions

Although phytoremediation has received considerable attention recently, and there are
an increasing number of reports suggesting that it should become the technology of choice
for the clean up of various types of environmental contamination, this technology is still in
its infancy and it has yet to be used commercially to any extent. Nevertheless, it is
predicted to account for approximately 10 – 15% of the growing environmental remedia-
tion market by the year 2010. However, to realize the full potential of this technology, it is
necessary for plants to grow as large as possible in the presence of various environmental
contaminants. One way to achieve this goal is to utilize plant growth-promoting bacteria to
facilitate the growth of the plants used for phytoremediation.

Acknowledgements

Work from the author’s laboratory was supported by grants from the Natural Science
and Engineering Research Council, CRESTech (a province of Ontario Centre of
392 B.R. Glick / Biotechnology Advances 21 (2003) 383–393

Excellence) and Inco. The following individuals contributed to the work reviewed here:
Genrich Burd, Donna Penrose, Varvara Grichko, Lin Nie, George Dixon, Wenbo Ma,
Bruce Greenberg, XiaoDong Huang, Jiping Li, Saleh Shah and Cheryl Patten.

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