AR242-PP56-02 ARI 1 April 2005 13:36

Phytoremediation
Elizabeth Pilon-Smits
Biology Department, Colorado State University, Fort Collins, Colorado 80523;
email: epsmits@[Link]

Annu. Rev. Plant Biol. Key Words

2005. 56:1539
pollution, decontamination, metals, organics, bioremediation
doi: 10.1146/
[Link].56.032604.144214
Abstract
Copyright

c 2005 by
Annual Reviews. All rights Phytoremediation, the use of plants and their associated microbes for
reserved environmental cleanup, has gained acceptance in the past 10 years as
First published online as a a cost-effective, noninvasive alternative or complementary technology
Review in Advance on for engineering-based remediation methods. Plants can be used for
January 11, 2005 pollutant stabilization, extraction, degradation, or volatilization. These
1543-5008/05/0602- different phytoremediation technologies are reviewed here, including
0015$20.00 their applicability for various organic and inorganic pollutants, and most
suitable plant species. To further enhance the efciency of phytoreme-
diation, there is a need for better knowledge of the processes that affect
pollutant availability, rhizosphere processes, pollutant uptake, translo-
cation, chelation, degradation, and volatilization. For each of these pro-
cesses I review what is known so far for inorganic and organic pollutants,
the remaining gaps in our knowledge, and the practical implications for
designing phytoremediation strategies. Transgenic approaches to en-
hance these processes are also reviewed and discussed.

15
AR242-PP56-02 ARI 1 April 2005 13:36

their properties, organics may be degraded in

Contents the root zone of plants or taken up, followed
by degradation, sequestration, or volatiliza-
INTRODUCTION . . . . . . . . . . . . . . . . . . 16
tion. Organic pollutants that have been
Phytoremediation: Advantages,
successfully phytoremediated include organic
Limitations, Present Status . . . . . . 16
solvents such as TCE (the most common pollu-
Phytoremediation Technologies and
tant of groundwater) (90, 111), herbicides such
Their Uses . . . . . . . . . . . . . . . . . . . . . 18
as atrazine (22), explosives such as TNT (61),
BIOLOGICAL PROCESSES
petroleum hydrocarbons such as oil, gasoline,
AFFECTING
benzene, toluene, and PAHs (4, 93, 110), the
PHYTOREMEDIATION . . . . . . . . . 21
fuel additive MTBE (26, 59, 128), and poly-
Pollutant Bioavailability . . . . . . . . . . . . 21
chlorinated biphenyls (PCBs) (53).
Rhizosphere Processes and
Inorganic pollutants occur as natural ele-
Remediation . . . . . . . . . . . . . . . . . . . . 22
ments in the earths crust or atmosphere, and
Plant Uptake . . . . . . . . . . . . . . . . . . . . . . 24
human activities such as mining, industry, traf-
Chelation and Compartmentation in
c, agriculture, and military activities promote
Roots . . . . . . . . . . . . . . . . . . . . . . . . . . 25
their release into the environment, leading to
Translocation . . . . . . . . . . . . . . . . . . . . . . 26
toxicity (91). Inorganics cannot be degraded,
Chelation and Compartmentation in
but they can be phytoremediated via stabiliza-
Leaves. . . . . . . . . . . . . . . . . . . . . . . . . . 27
tion or sequestration in harvestable plant tis-
Degradation . . . . . . . . . . . . . . . . . . . . . . . 28
sues. Inorganic pollutants that can be phytore-
Volatilization . . . . . . . . . . . . . . . . . . . . . . 29
mediated include plant macronutrients such as
NEW DEVELOPMENTS IN
nitrate and phosphate (60), plant trace elements
PHYTOREMEDIATION . . . . . . . . . 30
such as Cr, Cu, Fe, Mn, Mo, and Zn (76),
nonessential elements such as Cd, Co, F, Hg,
Se, Pb, V, and W (15, 60), and radioactive iso-
topes such as 238 U, 137 Cs, and 90 Sr (34, 35, 87).
INTRODUCTION Phytoremediation can be used for solid,
liquid, and gaseous substrates. Polluted soils
Phytoremediation: Advantages, and sediments have been phytoremediated at
Limitations, Present Status military sites (TNT, metals, organics), agri-
Phytoremediation is the use of plants and cultural elds (herbicides, pesticides, metals,
Phytoremediation:
the use of plants and their associated microbes for environmental selenium), industrial sites (organics, metals,
their associated cleanup (99, 107, 108). This technology makes arsenic), mine tailings (metals), and wood treat-
microbes for use of the naturally occurring processes by ment sites (PAHs) (8, 41, 93, 101, 129). Polluted
environmental cleanup which plants and their microbial rhizosphere waters that can be phytoremediated include
ora degrade and sequester organic and inor- sewage and municipal wastewater (nutrients,
TCE: ganic pollutants. Phytoremediation is an ef- metals), agricultural runoff/drainage water (fer-
trichloroethylene cient cleanup technology for a variety of tilizer nutrients, metals, arsenic, selenium,
TNT: trinitrotoluene organic and inorganic pollutants. Organic pol- boron, organic pesticides, and herbicides), in-
lutants in the environment are mostly man dustrial wastewater (metals, selenium), coal pile
PAH: polycyclic
aromatic hydrocarbon made and xenobiotic to organisms. Many of runoff (metals), landll leachate, mine drainage
them are toxic, some carcinogenic. Organic (metals), and groundwater plumes (organics,
MTBE: methyl
tertiary butyl ether pollutants are released into the environment metals) (38, 42, 52, 60, 74, 101). Plants can also
via spills (fuel, solvents), military activities (ex- be used to lter air, both outdoors and indoors,
plosives, chemical weapons), agriculture (pes- from, e.g., NOx , SO2 , ozone, CO2 , nerve gases,
ticides, herbicides), industry (chemical, petro- dust or soot particles, or halogenated volatile
chemical), wood treatment, etc. Depending on hydrocarbons (64, 86).

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Phytoremediation has gained popularity (Eastern Europe) await remediation. Phyto-

with government agencies and industry in the remediation may also become a technology of
past 10 years. This popularity is based in part choice for remediation projects in developing
on the relatively low cost of phytoremedia- countries because it is cost-efcient and easy to
tion, combined with the limited funds avail- implement.
able for environmental cleanup. The costs Phytoremediation has advantages but also
associated with environmental remediation are limitations. The plants that mediate the cleanup
staggering. Currently, $68 billion per year is have to be where the pollutant is and have to
spent for environmental cleanup in the United be able to act on it. Therefore, the soil prop-
States, and $2550 billion per year worldwide erties, toxicity level, and climate should allow
(47, 122). Because biological processes are ulti- plant growth. If soils are toxic, they may be
mately solar-driven, phytoremediation is on av- made more amenable to plant growth by adding
erage tenfold cheaper than engineering-based amendments, as described below. Phytoreme-
remediation methods such as soil excavation, diation is also limited by root depth because
soil washing or burning, or pump-and-treat the plants have to be able to reach the pollu-
systems (47). The fact that phytoremediation tant. Root depth is typically 50 cm for herba-
is usually carried out in situ contributes to ceous species or 3 m for trees, although cer-
its cost-effectiveness and may reduce exposure tain phreatophytes that tap into groundwater
of the polluted substrate to humans, wildlife, have been reported to reach depths of 15 m
and the environment. Phytoremediation also or more, especially in arid climates (88). The
enjoys popularity with the general public as limitations of root depth may be circumvented
a green clean alternative to chemical plants by deep planting of trees in boreholes (up
and bulldozers. Thus, government agencies like to 12 m) or pumping up polluted ground-
to include phytoremediation in their cleanup water for plant irrigation. Depending on the
strategies to stretch available funds, corpora- biological processes involved, phytoremedia-
tions (e.g., electric power, oil, chemical indus- tion may also be slower than the more es-
try) like to advertise their involvement with this tablished remediation methods like excavation,
environment-friendly technology, and environ- incineration, or pump-and-treat systems. Flow-
mental consultancy companies increasingly in- through phytoremediation systems and plant
clude phytoremediation in their package of degradation of pollutants work fairly fast (days
offered technologies. or months), but soil cleanup via plant accu-
The U.S. phytoremediation market now mulation often takes years, limiting applica-
comprises $100150 million per year, or 0.5% bility. Phytoremediation may also be limited
of the total remediation market (D. Glass, per- by the bioavailability of the pollutants. If only
sonal communication). For comparison, biore- a fraction of the pollutant is bioavailable, but
mediation (use of bacteria for environmental the regulatory cleanup standards require that
cleanup) comprises about 2% (47). Commer- all of the pollutant is removed, phytoremedia-
cial phytoremediation involves about 80% or- tion is not applicable by itself (43). Pollutant
ganic and 20% inorganic pollutants (D. Glass, bioavailability may be enhanced to some ex-
personal communication). The U.S. phytore- tent by adding soil amendments, as described
mediation market has growntwo- to three- below.
fold in the past 5 years, from $3049 million Nonbiological remediation technologies
in 1999 (47). In Europe there is no signi- and bio/phytoremediation are not mutually ex-
cant commercial use of phytoremediation, but clusive. Because pollutant distribution and con-
this may develop in the near future because centration are heterogeneous for many sites,
interest and funding for phytoremediation re- the most efcient and cost-effective remedia-
search are increasing rapidly, and many pol- tion solution may be a combination of different
luted sites in new European Union countries technologies, such as excavation of the most

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AR242-PP56-02 ARI 1 April 2005 13:36

Rhizofiltration: use
contaminated spots followed by polishing the Phytoremediation Technologies and
of plants in site with the use of plants. Such an integrated Their Uses
hydroponic setup for remediation effort requires a multidisciplinary
Plants and their rhizosphere organisms can be
ltering polluted water team of knowledgeable scientists.
used for phytoremediation in different ways
Phytoextraction: use This review aims to give a broad overview
(see Figure 1). They can be used as lters in
of plants to clean up of the state of the science of phytoremedia-
constructed wetlands (60) or in a hydroponic
pollutants via tion, with references to other publications that
accumulation in setup (100); the latter is called rhizoltration.
give more in-depth information. After an intro-
harvestable tissues Trees can be used as a hydraulic barrier to
duction to the various phytoremediation tech-
create an upward water ow in the root zone,
nologies, the plant processes involved in uptake,
preventing contamination to leach down, or
translocation, sequestration, and degradation of
to prevent a contaminated groundwater plume
organic and inorganic pollutants are reviewed
from spreading horizontally (90). The term
in the context of phytoremediation. Finally,
phytostabilization denotes the use of plants to
new developments including genetic engineer-
stabilize pollutants in soil (13), either simply by
ing are discussed with respect to their prospects
preventing erosion, leaching, or runoff, or by
for phytoremediation.
converting pollutants to less bioavailable forms

Figure 1
Phytoremediation
technologies used for
remediating polluted
water, soil, or air. The
red circles represent
the pollutant.

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AR242-PP56-02 ARI 1 April 2005 13:36

(e.g., via precipitation in the rhizosphere).

Plants can also be used to extract pollutants and
accumulate them in their tissues, followed by
harvesting of the (above ground) plant material.
This technology is called phytoextraction (15).
The plant material can subsequently be used
for nonfood purposes (e.g., wood, cardboard)
or ashed, followed by disposal in a landll or,
in the case of valuable metals, recycling of the
accumulated element. The latter is termed phy-
tomining (23).
Plants can facilitate biodegradation of
organic pollutants by microbes in their rhizo-
sphere (see Figure 2). This is called phytostim-
ulation or rhizodegradation (82). Plants can also
degrade organic pollutants directly via their
own enzymatic activities, a process called phy-
todegradation (82). After uptake in plant tissue,
certain pollutants can leave the plant in volatile
form; this is called phytovolatilization (118). buffer strips to intercept horizontal migration Figure 2
These various phytoremediation technologies of polluted ground water plumes and redirect Possible fates of
pollutants during
are not mutually exclusive; for instance, in a water ow upward (82). Natural attenuation is
phytoremediation: the
constructed wetland, accumulation, stabiliza- suitable for remote areas with little human use pollutant (represented
tion and volatilization can occur simultaneously and relatively low levels of contamination. Phy- by red circles) can be
(52). Because the processes involved in phy- toextraction is mainly used for metals and other stabilized or degraded
toremediation occur naturally, vegetated pol- toxic inorganics (Se, As, radionuclides) (9, 15). in the rhizosphere,
sequestered or
luted sites have a tendency to clean themselves Phytostimulation is used for hydrophobic or-
degraded inside the
up without human interference. This so-called ganics that cannot be taken up by plants but plant tissue, or
natural attenuation is the simplest form of phy- that can be degraded by microbes. Examples volatilized.
toremediation and involves only monitoring. are PCBs, PAHs, and other petroleum hydro-
The different phytoremediation technolo- carbons (62, 93). Phytodegradation works well
gies described above are suitable for different for organics that are mobile in plants such as Rhizodegradation/
classes of pollutants. Constructed wetlands have herbicides, TNT, MTBE, and TCE (21, 128). phytostimulation:
degradation of
been used for a wide range of inorganics includ- Phytovolatilization can be used for VOCs (128) pollutants in the
ing metals, Se, perchlorate, cyanide, nitrate, and such as TCE and MTBE, and for a few inor- rhizosphere due to
phosphate (52, 60, 92), as well as certain organ- ganics that can exist in volatile form, i.e., Se and microbial activity
ics such as explosives and herbicides (60, 63, Hg (52, 105). Phytodegradation:
83, 110). Rhizoltration in an indoor, contained Different phytotechnologies make use of breakdown of
setup is relatively expensive to implement different plant properties and typically differ- pollutants by plant
and therefore most useful for relatively small ent plant species are used for each. Favorable enzymes, usually
inside tissues
volumes of wastewater containing hazardous plant properties for phytoremediation in gen-
inorganics such as radionuclides (35, 87). The eral are to be fast growing, high biomass, com- Phytovolatilization:
release of pollutants by
principle of phytostabilization is used, e.g., petitive, hardy, and tolerant to pollution. In plants in volatile form
when vegetative caps are planted on sites con- addition, high levels of plant uptake, translo-
VOC: volatile organic
taining organic or inorganic pollutants, or when cation, and accumulation in harvestable tissues compound
trees are used as hydraulic barriers to prevent are important properties for phytoextraction
PCB: polychlorinated
leaching or runoff of organic or inorganic con- of inorganics. Favorable plant properties for biphenyl
taminants. Trees can also be used in so-called phytodegradation are large, dense root systems

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and high levels of degrading enzymes. A large growth, high biomass, and high tolerance and
root surface area also favors phytostimulation, accumulation of metals and other inorganics
as it promotes microbial growth; furthermore, (15, 107). A special category of plants are the
production of specic exudate compounds may so-called hyperaccumulators: plant species that
further promote rhizodegradation via specic accumulate one or more inorganic elements to
plant-microbe interactions (93). levels 100-fold higher than other species grown
In constructed wetlands for phytoremedi- under the same conditions (19). Hyperaccumu-
ation, a variety of emergent, submerged, and lators have been reported for As, Co, Cu, Mn,
oating aquatic species are used. Popular gen- Ni, Pb, Se, and Zn (7, 11, 77). These elements
era/species are cattail (Typha sp.), parrot feather are typically hyperaccumulated up to 0.11% of
(Myriophyllum sp.), Elodea sp., Azolla sp., duck- dry weight even from low external concentra-
weed (Lemna sp.), water hyacinth (Eichhornia tions. Despite these properties hyperaccumula-
crassipes), and Spartina sp. Poplar (Populus sp.) tors are not very popular for phytoremediation
and willow (Salix sp.) can be used on the edges because they are often slow growing and attain
of wetlands. For brackish water, certain species low biomass. So far only one hyperaccumula-
of Spartina are useful, as well as pickleweed (Sal- tor species, the Ni hyperaccumulator Alyssum
icornia sp.) and saltgrass (Distichlis spicata) (74). bertolonii, has been used for phytoremediation
For inorganics, the oating species water hy- in the eld (23, 73). The recently discovered
acinth, Azolla, and duckweed are popular be- As hyperaccumulating fern Pteris vittata may
cause they are good metal accumulators and also show promise for phytoextraction of As
can be harvested easily; cattail and poplar are (77).
also used because they are tolerant, grow fast, For phytostimulation of microbial degraders
and attain a high biomass. Aquatic plants that in the root zone, grasses such as fescue (Fes-
work well for organics remediation include par- tuca sp.), ryegrass (Lolium sp.), Panicum sp., and
rot feather and Elodea (83) because they have prairie grasses (e.g., Buchloe dactyloides, Bouteloua
high levels of organic-degrading enzymes. Rhi- sp.) are popular because they have very dense
zoltration involves aeration and therefore is and relatively deep root systems and thus a large
not limited to aquatic species; it often makes use root surface area (4). Mulberry trees also enjoy
of terrestrial species with large roots and good popularity for use in phytostimulation because
capacity to accumulate inorganics, such as sun- of their reported ability to produce phenolic
ower (Helianthus annuus) or Indian mustard compounds that stimulate expression of micro-
(Brassica juncea) (35). bial genes involved in PCB and PAH degra-
In a vegetative cap for phytostabilization, a dation (44, 72, 93). For phytodegradation of
combination of trees and grasses may be used. TCE and atrazine, poplar has been the most
Fast-transpiring trees such as poplar maintain popular and efcient species so far, owing to its
an upward ow to prevent downward leaching, high transpiration rate and capacity to degrade
while grasses prevent wind erosion and lateral and/or volatilize these pollutants (22, 110).
runoff with their dense root systems. Grasses Poplar is also the most-used species for phy-
tend to not accumulate inorganic pollutants in tovolatilization of VOCs because of its high
their shoots as much as dicot species (12), min- transpiration rate, which facilitates the move-
imizing exposure of wildlife to toxic elements. ment of these compounds through the plant
Poplar trees are very efcient at intercepting into the atmosphere. For volatilization of in-
horizontal groundwater plumes and redirect- organics, only Se has been investigated in de-
ing water ow upward because they are deep tail. In general, plant species that take up and
rooted and transpire at very high rates, creat- volatilize sulfur compounds also accumulate
ing a powerful upward ow (27, 82). and volatilize Se well because S and Se are
Popular species for phytoextraction are In- chemically similar and their metabolism oc-
dian mustard and sunower because of their fast curs via the same pathways (2). Members of the

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AR242-PP56-02 ARI 1 April 2005 13:36

Brassica genus are particularly good volatilizers tant, soil properties, environmental conditions,
CEC: cation
of Se (117). Among the aquatic species tested, and biological activity. Soils with small parti- exchange capacity
rice, rabbitfoot grass, Azolla, and pickleweed cle size (clay) hold more water than sandy soils,
were the best Se volatilizers (52, 74, 97, 133). and have more binding sites for ions, especially
Finally, when choosing plant species for a cations (CEC) (116). The concentration of or-
certain site, it is advisable to include species that ganic matter (humus) in the soil is also positively
grow locally on or near the site. These species correlated with CEC, as well as with the ca-
are competitive under the local conditions and, pacity to bind hydrophobic organic pollutants.
if they are growing on the site, can tolerate the This is because humus mainly consists of dead
pollutant. plant material, and plant cell walls have nega-
tively charged groups that bind cations, as well
as lignin that binds hydrophobic compounds
BIOLOGICAL PROCESSES (21).
AFFECTING Two important chemical properties of a pol-
PHYTOREMEDIATION Log Kow : the octanol:
lutant that affect its movement in soils are hy- water distribution
Phytoremediation effectively removes pollu- drophobicity and volatility. Hydrophobicity is coefcient, a measure
tants, but in many cases the underlying bio- usually expressed as the octanol:water partition for pollutant
logical mechanisms remain largely unknown. coefcient, or log Kow (121). A high log Kow hydrophobicity
To increase the efciency of phytoremediation corresponds with high hydrophobicity. Ex-
technologies, it is important that we learn more tremely hydrophobic molecules such as PCBs, DNAPL: dense
about the biological processes involved. These PAHs, and other hydrocarbons (log Kow > 3) nonaqueous phase
include plant-microbe interactions and other are tightly bound to soil organic matter and do liquid
rhizosphere processes, plant uptake, translo- not dissolve in the soil pore water. This lack of LNAPL: light
cation mechanisms, tolerance mechanisms bioavailability limits their ability to be phytore- nonaqueous phase
(compartmentation, degradation), and plant mediated, leading to their classication as re- liquid
chelators involved in storage and transport. calcitrant pollutants. Nonaqueous liquids may
Other processes that need more study are sink down to the ground water and, depending
movement of pollutants through ecosystems via on whether they are more or less dense than
the soil-water-plant system to higher trophic water, end up below the aquifer (DNAPLs) or
levels. In the following sections we follow the on top of the aquifer (LNAPLs). Organics with
path of pollutants toward, into, and within the moderate to high water solubility (log Kow < 3)
plant during phytoremediation. For each step will be able to migrate in the soil pore water to
I discuss what is known and not known about an extent that is inversely correlated with their
factors inuencing remediation, potential lim- log Kow .
iting steps for organic and inorganic pollutants, Pollutant volatility, expressed as Henrys law
and the practical implications for phytoremedi- constant (Hi ), is a measure of a compounds ten-
ation. Also, I discuss transgenic approaches that dency to partition to air relative to water (26).
have been or may be used to enhance phytore- Pollutants with Hi > 104 tend to move in the
mediation efciency at each step. air spaces between soil particles, whereas pol-
lutants with Hi < 106 move predominantly
in water. If Hi is between 104 and 106 ,
Pollutant Bioavailability compounds are mobile in both air and water.
For plants and their associated microbes to re- Both water-mobile and air-mobile organic con-
mediate pollutants, they must be in contact taminants can diffuse passively through plants.
with them and able to act on them. Therefore, While the fate of water-mobile organics is phy-
the bioavailability of a pollutant is important todegradation or sequestration, volatile organ-
for its remediation. Pollutant bioavailability de- ics can be rapidly volatilized by plants without
pends on the chemical properties of the pollu- chemical modication (18).

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Inorganics are usually present as charged phytoextraction where EDTA is added to soil
cations or anions, and thus are hydrophilic. The shortly before plant harvesting, greatly increas-
bioavailability of cations is inversely correlated ing plant metal uptake (108). Before chelate-
with soil CEC. At lower soil pH, the bioavail- assisted phytoextraction is used in the eld, it is
ability of cations generally increases due to re- important to do a risk assessment study to de-
placement of cations on soil CEC sites by H+ termine possible effects of the chelator on metal
ions (116). The bioavailability of ions is also af- leaching. In other situations it may be desirable
fected by the redox conditions. Most terrestrial to decrease metal bioavailability if metals are
soils have oxidizing conditions, and elements present at phytotoxic levels or in phytostabiliza-
that can exist in different oxidation states will be tion. In such cases lime may be mixed in with
in their most oxidized form [e.g., as selenate, ar- the soil to increase the pH or organic matter to
senate, Cr(VI), Fe3+ ]. In aquatic habitats more bind metals (12, 20). Adding organic matter also
reducing conditions exist, which favor more re- decreases the bioavailability of hydrophobic or-
duced elemental forms [e.g., selenite, arsenite, ganics, whereas adding surfactants (soap) may
Cr(III), Fe2+ ]. The oxidation state of an ele- increase their bioavailability. For organics that
ment may affect its bioavailability (e.g., its sol- can exist in more or less protonated forms with
ubility), its ability to be taken up by plants, as different charges, manipulation of soil pH can
well as its toxicity. Other physical conditions also affect their solubility and ability to move
that affect pollutant migration and bioavail- into plants. Finally, water supply may be op-
ability are temperature and moisture. Higher timized to facilitate pollutant migration while
temperatures accelerate physical, chemical, and preventing leaching or runoff.
biological processes in general. Precipitation
will stimulate general plant growth, and higher
soil moisture will increase migration of water- Rhizosphere Processes and
soluble pollutants. The bioavailability of pollu- Remediation
tants may also be altered by biological activities, Rhizosphere remediation occurs completely
as described in the next section. In polluted soils without plant uptake of the pollutant in the
the more bioavailable (fraction of ) pollutants area around the root. The rhizosphere extends
tend to decrease in concentration over time approximately 1 mm around the root and is
due to physical, chemical, and biological pro- under the inuence of the plant. Plants re-
cesses, leaving the less or nonbioavailable (frac- lease a variety of photosynthesis-derived or-
tion of ) pollutants. Consequently, pollutants in ganic compounds in the rhizosphere that can
aged polluted soils tend to be less bioavailable serve as carbon sources for heterotrophic fungi
and more recalcitrant than pollutants in soil that and bacteria (16). As much as 20% of carbon
is newly contaminated, making aged soils more xed by a plant may be released from its roots
difcult to phytoremediate (93). (93). As a result, microbial densities are 1
Understanding the processes affecting pol- 4 orders of magnitude higher in rhizosphere
EDTA: ethylene
diamine tetra acetic lutant bioavailabilty can help optimize phy- soil than in bulk soil, the so-called general
acid toremediation efciency. Amendments may be rhizosphere effect (108). In turn, rhizosphere
added to soil that make metal cations more microbes can promote plant health by stimulat-
bioavailable for plant uptake. For instance, ing root growth (some microorganisms produce
adding the natural organic acids citrate or plant growth regulators), enhancing water and
malate will lower the pH and chelate metals mineral uptake, and inhibiting growth of other,
such as Cd, Pb, and U from soil particles, usu- NO pathogenic soil microbes (65).
ally making them more available for plant up- In rhizosphere remediation it is often dif-
take. The synthetic metal chelator EDTA is cult to distinguish to what extent effects are
also extremely efcient at releasing metals from due to the plant or to the rhizosphere mi-
soil. This principle is used in chelate-assisted crobes. Laboratory studies with sterile plants

22 Pilon-Smits
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and microbial isolates can be used to address rhizosphere by microbial activity include PAHs,
this question. Rhizosphere remediation may be PCBs, and petroleum hydrocarbons (62, 93).
a passive process. Pollutants can be phytosta- Plants can stimulate these microbial degrada-
bilized simply via erosion prevention and hy- tion processes. First, plant carbon compounds
draulic control as described above. There is also released into the rhizosphere facilitate a higher
passive adsorption of organic pollutants and in- microbial densitythe general rhizosphere ef-
organic cations to the plant surface. Adsorp- fect. Second, secondary plant compounds re-
tion of lipophilic organics to lignin groups in leased from roots may specically induce
the cell walls is called lignication (82). Rhi- microbial genes involved in degradation of the
zosphere remediation may also be the result organic compound, or act as a cometabolite
of active processes mediated by plants and/or to facilitate microbial degradation (44, 72, 93).
microbes. These processes may affect pollutant Better knowledge of these plant-microbe in-
bioavailability, uptake, or degradation. teractions is needed to more efciently design
Pollutant bioavailability may be affected by phytoremediation strategies or engineer more
various plant and/or microbial activities. Some efcient plant-microbe consortia.
bacteria are known to release biosurfactants Rhizosphere processes that favor phy-
(e.g., rhamnolipids) that make hydrophobic toremedation may be optimized by the choice
pollutants more water soluble (126). Plant exu- of plant species, e.g., plants with large and dense
dates or lysates may also contain lipophilic com- root systems for phytostimulation, or aquatic
pounds that increase pollutant water solubility plants for metal precipitation. If a certain ex-
or promote biosurfactant-producing microbial udate compound is identied to enhance phy-
populations (113). Furthermore, plant- and toremediation (e.g., a chelator or a secondary
microbe-derived enzymes can affect the solu- metabolite that stimulates microbial degrada-
bility and thus the bioavailability of organic pol- tion) plants can be selected or genetically en-
lutants via modication of side groups (131). gineered to produce large amounts of this
Bioavailability of metals may be enhanced compound. In one such study, overexpression of
by metal chelators that are released by plants citrate synthase in plants conferred enhanced
and bacteria. Chelators such as siderophores, aluminum tolerance, probably via enhanced cit-
organic acids, and phenolics can release metal rate release into the rhizosphere, which pre-
cations from soil particles. This usually makes vented Al uptake due to complexation (28). In
the metals more available for plant uptake (116) another approach to stimulate rhizosphere re-
although in some cases it can prevent up- mediation, certain agronomic treatments may
take (28). Furthermore, plants extrude H+ via be employed that favor the production of gen-
ATPases, which replace cations at soil CEC eral and specic exudate compounds, such as
sites, making metal cations more bioavailable clipping or fertilization (72). Inorganic fertil-
(116). Some plant roots release oxygen, such as izer is preferred over organic fertilizer (manure)
aquatic plants that have aerenchyma (air chan- for use in phytostimulation because the latter
nels in the stem that allow oxygen to diffuse to provides an easy-to-digest carbon source that
the root); this can lead to the oxidation of metals microbes may prefer to use instead of the or-
to insoluble forms (e.g., FeO3 ) that precipitate ganic pollutant.
on the root surface (60). Conversely, enzymes If the microbial consortia responsible for the
on the root surface may reduce inorganic pollu- remediation process are known, it may be pos-
tants, which may affect their bioavailability and sible to increase the abundance of these species
toxicity (e.g., CrVI to CrIII) (76). by the choice of vegetation. An alternative ap-
Organic pollutants may be degraded in the proach is to grow these microbial isolates in
rhizosphere by root-released plant enzymes or large amounts and add them to the soil, a pro-
via phytostimulation of microbial degradation. cess called bioaugmentation. Introducing non-
Examples of organics that are degraded in the native microbes to sites is considered ineffective

[Link] Phytoremediation 23
AR242-PP56-02 ARI 1 April 2005 13:36

because they tend to be outcompeted by the to nutrients and are taken up inadvertently (e.g.,
established microbial populations. In another arsenate is taken up by phosphate transporters,
approach to optimize rhizosphere remediation, selenate by sulfate transporters) (1, 112). In-
the watering regime may be regulated to pro- organics usually exist as ions and cannot pass
vide an optimal soil moisture for plant and mi- membranes without the aid of membrane trans-
crobial growth. If redox reactions are involved porter proteins. Because uptake of inorganics
in the remediation process, periodic ooding depends on a discrete number of membrane
and draining of constructed wetlands may be ef- proteins, their uptake is saturable, following
fective to alternate reducing and oxidizing con- Michaelis Menten kinetics (80). For most ele-
ditions (62). ments multiple transporters exist in plants. The
model plant Arabidopsis thaliana, for instance,
has 150 different cation transporters (6), and
Plant Uptake 14 transporters for sulfate alone (56). Individ-
Uptake of pollutants by plant roots is differ- ual transporter proteins have unique proper-
Root concentration
factor (RCF): the ent for organics and inorganics. Organic pol- ties with respect to transport rate, substrate
ratio of pollutant lutants are usually manmade, and xenobiotic to afnity, and substrate specicity (low afnity
concentration in root the plant. As a consequence, there are no trans- transporters tend to be more promiscuous) (80).
relative to external porters for these compounds in plant mem- These properties may be subject to regulation
solution, used as a
branes. Organic pollutants therefore tend to by metabolite levels or regulatory proteins (e.g.,
measure for plant
uptake move into and within plant tissues driven by kinases). Furthermore, the abundance of each
simple diffusion, dependent on their chemi- transporter varies with tissue-type and envi-
cal properties. An important property of the ronmental conditions, which may be regulated
organic pollutant for plant uptake is its hy- at the transcription level or via endocytosis.
drophobicity (17, 121). Organics with a log Kow As a consequence, uptake and movement of
between 0.5 and 3 are hydrophobic enough to inorganics in plants are complex species- and
move through the lipid bilayer of membranes, conditions-dependent processes, and difcult
and still water soluble enough to travel into to capture in a model.
the cell uids. If organics are too hydrophilic When inorganic pollutants accumulate in
(log Kow < 0.5) they cannot pass membranes tissues they often cause toxicity, both directly
and never get into the plant; if they are too hy- by damaging cell structure (e.g., by causing ox-
drophobic (log Kow > 3) they get stuck in mem- idative stress due to their redox activity) and
branes and cell walls in the periphery of the indirectly via replacement of other essential nu-
plant and cannot enter the cell uids. Because trients (116). Organics tend to be less toxic to
the movement of organics into and through plants, partly because they are not accumulated
plants is a physical rather than biological pro- as readily and because they tend to be less re-
cess, it is fairly predictable across plant species active. Thus, when soils are polluted with a
and lends itself well to modeling (26). The ten- mixture of organics and metals the inorgan-
dency of organic pollutants to move into plant ics are most likely to limit plant growth and
roots from an external solution is expressed as phytoremediation. Phytoremediation of mixed
the root concentration factor (RCF = equilib- pollutants (organics and inorganics) is an un-
rium concentration in roots/equilibrium con- derstudied area, but very relevant because many
centratrion in external solution). sites contain mixed pollution.
In contrast, inorganics are taken up by bi- The presence of rhizosphere microbes can
ological processes via membrane transporter affect plant uptake of inorganics. For instance,
proteins. These transporters occur naturally mycorrhizal fungi can both enhance uptake of
because inorganic pollutants are either nutri- essential metals when metal levels are low and
ents themselves (e.g., nitrate, phosphate, cop- decrease plant metal uptake when metals are
per, manganese, zinc) or are chemically similar present at phytotoxic levels (46, 104). Also,

24 Pilon-Smits
AR242-PP56-02 ARI 1 April 2005 13:36

rhizosphere bacteria can enhance plant uptake thermore, altering plant production of chela-
of mercury and selenium (29). The mechanisms tor molecules can affect plant metal accumula-
of these plant-microbe interactions are still tion (39, 49, 54, 134, 135). Hyperaccumulator
largely unclear; microbe-mediated enhanced species offer potentially interesting genetic ma-
plant uptake may be due to a stimulatory ef- terial to be transferred to high-biomass species.
fect on root growth, microbial production of Constitutive expression of a Zn transporter in
metabolites that affect plant gene expression the root cell membrane is one of the underlying
of transporter proteins, or microbial effects on mechanisms of the natural Zn hyperaccumula-
bioavailability of the element (30). tor Thlaspi caerulescens (94). Research is ongoing
Depending on the phytoremediation strat- to isolate genes involved in metal hyperaccumu-
egy, pollutant uptake into the plant may be de- lation and hypertolerance.
sirable (e.g., for phytoextraction) or not (e.g.,
for phytostabilization). For either application,
plant species with the desired properties may Chelation and Compartmentation
be selected. Screening studies under uniform in Roots
conditions are a useful strategy to compare As mentioned above, plants can release com- GSH: glutathione
uptake characteristics of different species for pounds from their roots that affect pollutant
PC: phytochelatin
different pollutants. Agronomic practices may solubility and uptake by the plant. Inside plant
MT: metallothionein
also be employed to maximize pollutant uptake. tissues such chelator compounds also play a role
protein
Plant species may be selected for suitable root- in tolerance, sequestration, and transport of in-
ing depth and root morphology (88). Further- organics and organics (103). Phytosiderophores
more, plant roots can be guided to grow into are chelators that facilitate uptake of Fe and per-
the polluted zone via deep planting in a cas- haps other metals in grasses; they are biosynthe-
ing, forcing the roots to grow downward into sized from nicotianamine, which is composed
the polluted soil and to tap into polluted water of three methionines coupled via nonpeptide
rather than rainwater (88). Supplemental water bonds (57). Nicotianamine also chelates met-
(via irrigation) and oxygen (via air tube to roots) als and may facilitate their transport (115, 127).
may also facilitate pollutant uptake, and soil nu- Organic acids (e.g., citrate, malate, histidine)
trient levels may be optimized by fertilization. not only can facilitate uptake of metals into
Not only will nutrients promote plant growth roots but also play a role in transport, seques-
and thus uptake of the pollutant, they may also tration, and tolerance of metals (70, 107, 127).
affect plant uptake of pollutants via ion compe- Metals can also be bound by the thiol-rich pep-
tition at the soil and plant level. For instance, tides GSH and PCs, or by the Cys-rich MTs
supplying phosphate will release arsenate from (24). Chelated metals in roots may be stored
soils, making it more bioavailable; on the other in the vacuole or exported to the shoot via the
hand, phosphate will compete with arsenate for xylem. As described in more detail below, or-
uptake by plants because both are taken up by ganics may be conjugated and stored or de-
phosphate transporters (1). graded enzymatically. An overview of these pro-
It may also be possible to manipulate plant cesses is depicted in Figure 3.
accumulation by genetic engineering. A trans- Chelation in roots can affect phytoremedi-
genic approach that may be used to alter up- ation efciency as it may facilitate root seques-
take of inorganic pollutants is overexpression tration, translocation, and/or tolerance. Root
or knockdown of membrane transporter pro- sequestration may be desirable for phytostabi-
teins. This approach was used successfully to lization (less exposure to wildlife) whereas ex-
enhance accumulation of Ca, Cd, Mn, Pb, and port to xylem is desirable for phytoextraction.
Zn (5, 58, 123). The specicity of membrane If chelation is desirable, it may be enhanced
transporters for different inorganics may also be by selection or engineering of plants with
manipulated via protein engineering (102). Fur- higher levels of the chelator in question. Root

[Link] Phytoremediation 25
AR242-PP56-02 ARI 1 April 2005 13:36

Figure 3
Tolerance mechanisms
for inorganic and
organic pollutants in
plant cells.
Detoxication
generally involves
conjugation followed
by active sequestration
in the vacuole and
apoplast, where the
pollutant can do the
least harm. Chelators
shown are
GSH: glutathione,
Glu: glucose,
MT: metallothioneins,
NA: nicotianamine,
OA: organic acids,
PC: phytochelatins.
Active transporters are
shown as boxes with
arrows.

sequestration and export to xylem might be owing straight from the soil solution or root
manipulated by overexpression or knockdown apoplast into the root xylem (116). Organic pol-
of the respective membrane transporters in- lutants pass the membrane between root sym-
volved. Unfortunately, little is known about plast and xylem apoplast via simple diffusion.
these tissue-specic transporters of inorganics. The TSCF is the ratio of the concentration of
The completion of the sequencing of the Ara- a compound in the xylem uid relative to the
bidopsis and rice genomes should accelerate the external solution, and is a measure of uptake
analysis of transporter gene families. into the plant shoot. Entry of organic pollu-
Transpiration stream tants into the xylem depends on similar pas-
concentration factor
sive movement over membranes as their uptake
(TSCF): the ratio of Translocation
pollutant into the plant. Thus, the TSCF for organics
concentration in xylem Translocation from root to shoot rst requires shows a similar correlation with hydrophobic-
uid relative to a membrane transport step from root sym- ity as RCF: Compounds with a log Kow be-
external solution, used plast into xylem apoplast. The impermeable tween 0.5 and 3 are most easily transported
as a measure for plant
suberin layer in the cell wall of the root endo- to the xylem and translocated to the shoot
translocation
dermis (Casparian strip) prevents solutes from (121).

26 Pilon-Smits
AR242-PP56-02 ARI 1 April 2005 13:36

Inorganics require membrane transporter the pollutant, as discussed above. Once inside
proteins to be exported from the root endo- the leaf symplast, the pollutant may be compart-
dermis into the root xylem. Some inorganics mentized in certain tissues or cellular locations.
are chelated during xylem transport by organic In general, toxic pollutants are sequestered in
acids (histidine, malate, citrate), nicotianamine, places where they can do the least harm to es-
or thiol-rich peptides (67, 95, 115, 127). For sential cellular processes. At the cellular level,
most inorganics it is still unclear via which pollutants are generally accumulated in the vac-
transporter proteins they are exported to the uole or cell wall (21, 24). At the tissue level they
root xylem and to whichif anychelators may be accumulated in the epidermis and tri-
they are bound during transport. Better knowl- chomes (50, 69).
edge of the transporters and chelators involved When pollutants are sequestered in tissues,
XAS: X-ray
in translocation of inorganics would facilitate they are often bound by chelators or form con- absorption
the development of transgenics with more ef- jugates (see Figure 3). Toxic inorganics are usu- spectroscopy
cient phytoextraction capacity. ally metals. Chelators that are involved in metal
Bulk ow in the xylem from root to shoot is sequestration include the tripeptide GSH ( -
driven by transpiration from the shoot, which glu-cys-gly) and its oligomers, the PCs. XAS
creates a negative pressure in the xylem that has shown that inorganics that were complexed
pulls up water and solutes (116). Plant tran- by PCs in vivo include Cd and As (95); there
spiration depends on plant properties and en- may be others since PC synthesis is induced
vironmental conditions. Plant species differ by various other metals (24). After chelation
in transpiration rate, due to metabolic differ- by GSH or PCs, an ABC-type transporter ac-
ences (e.g., C3/C4/CAM photosynthetic path- tively transports the metal-chelate complex to
way) and anatomical differences (e.g., surface to the vacuole, where it is further complexed by
volume ratio, stomatal density, rooting depth) sulde (24, 75). Organic acids such as malate
(116). Species such as poplar are phreatophytes, and citrate are also likely metal (e.g., Zn) chela-
or water spenders; they have long roots that tors in vacuoles, as judged from XAS (70). Fer-
tap into the ground water (27). Mature poplar ritin is an iron chelator in chloroplasts (120).
trees can transpire 2001000 liters of water per Additional metal-chelating proteins exist (e.g.,
day (38, 132). In addition to plant species com- MTs) that may play a role in sequestration and
position, vegetation height and density affect tolerance (e.g., of Cu) and/or in homeostasis
transpiration, as well as environmental con- of essential metals (48). There is still much to
ditions: Transpiration is generally maximal at be discovered about the roles of these differ-
high temperature, moderate wind, low rela- ent chelators in transport and detoxication of
tive air humidity, and high light (116). Con- inorganic pollutants.
sequently, phytoremediation mechanisms that Conjugation to GSH also plays a role in
GST:
rely on translocation and volatilization are most sequestration and tolerance of organic pol- GSH-S-transferases
effective in climates with low relative humidity lutants (78). A large family of GSTs with
and high evapotranspiration. different substrate specicities mediate conju-
gation of organics to GSH in the cytosol (55, 68,
89). The glutathione S-conjugates are actively
Chelation and Compartmentation transported to the vacuole or the apoplast by
in Leaves ATP-dependent membrane pumps (79, 81, 109,
Import into leaf cells from leaf xylem involves 130). An alternative conjugation-sequestration
another membrane transport step. Inorganics mechanism for organics in plants involves cou-
are taken up by specic membrane transporter pling a glucose or a malonyl-group to the
proteins. Organics enter the leaf symplast from organic compound, followed by transport of
the shoot xylem by simple diffusion, the rate of the conjugate to the vacuole or the apoplast
which depends on the chemical properties of (25). These conjugation steps are mediated by

[Link] Phytoremediation 27
AR242-PP56-02 ARI 1 April 2005 13:36

a family of glucosyltransferases and malonyl- or chelator proteins (49, 54, 134, 135). In ad-
transferases, and the transport steps by ATP- dition, enzymes that couple the chelator or
dependent pumps (21). conjugant to the pollutant (GSH transferases,
To be conjugated, the organic compound glucosyltransferases) may be overexpressed (40)
may need chemical modication to create or enzymes that modify organics to make them
suitable side groups for conjugation. These amenable to conjugation (32, 33, 51).
modication reactions can be oxidative or In all cases where potentially toxic pollutants
reductive. For example, cytochrome P450 are accumulated in plant tissues, phytoremedi-
monooxygenases catalyze an oxidative transfor- ation in the eld should include a risk assess-
mation, incorporating an O atom from oxygen ment study because the plant material may pose
into an organic molecule such as atrazine to a threat to wildlife. The degree of toxicity will
create a hydroxyl side group (25). Nitroreduc- depend on leaf concentration but also on the
tases are an example of enzymes that mediate form of the pollutant that is accumulated. Dur-
a reductive transformation, converting a nitro ing accumulation the toxicity of the pollutant
group of, e.g., TNT to an amino group (83). may change. To test the potential toxicity of the
Other enzymes that mediate modications of plant material, a laboratory digestibility study
organic pollutants include dioxygenases, per- may be done using model organisms or in vitro
oxidases, peroxygenases, and carboxylesterases simulations of animal digestion systems. In the
(21). Thus, accumulation of organic pol- eld, exposure to wildlife may be minimized by,
lutants typically comprises three phases: e.g., fencing, netting, noise, and scarecrows.
chemical modication, conjugation, and se-
questration (Figure 3). This sequence of events
has been summarized as the green liver model Degradation
because of its similarity to mammalian detox- Only organic pollutants can be phytoremedi-
ication mechanisms (21, 109). Some natu- ated via degradation. Inorganic elements are
ral functions of the enzymes and transporters undegradable and can only be stabilized or
involved are to biosynthesize and transport moved and stored. In phytodegradation plant
natural plant compounds such as avonoids, enzymes act on organic pollutants and catabo-
alkaloids, and plant hormones, and to defend lize them, either mineralizing them completely
against biotic stresses (78, 98). to inorganic compounds (e.g., carbon diox-
Uptake and accumulation in leaves with- ide, water and Cl2 ), or degrading them par-
out toxic effects are desirable properties for tially to a stable intermediate that is stored in
phytoextraction. To maximize these processes, the plant (82). This enzymatic degradation of
plants may be selected or engineered that have organics can happen in both root and shoot tis-
higher levels of transporters involved in uptake sue. Degradation within plant tissues is gener-
of an inorganic pollutant from the xylem into ally attributed to the plant, but may in some
the leaf symplast. Better knowledge of the trans- cases involve endophytic microorganisms (10).
porters involved in the process would be help- Phytodegradation involves some of the same
ful because this is still a largely unexplored area. classes of enzymes responsible for accumulation
Similarly, plants with high transporter activities in tissues. The modifying enzymes that create
from cytosol to vacuole can be more efcient at side groups on organics that increase solubil-
storing toxic inorganics (58, 114, 123). Seques- ity and enable conjugation also play a role in
tration and tolerance may also be enhanced by the initial steps of phytodegradation. Thus, en-
selection or engineering of plants with higher zyme classes involved in phytodegradation in-
production of leaf chelators or conjugates. This clude dehalogenases, mono- and dioxygenases,
can be mediated by higher levels of enzymes peroxidases, peroxygenases, carboxylesterases,
that produce these conjugates, e.g., enzymes laccases, nitrilases, phosphatases, and nitrore-
synthesizing GSH, PCs, glucose, organic acids, ductases (131). Also, if pollutants are only

28 Pilon-Smits
AR242-PP56-02 ARI 1 April 2005 13:36

partially degraded and the degradation prod- methylated to form dimethylselenide (DMSe),
ucts stored in plants, these are often conjugated which is volatile (119). Volatilization of the in-
and sequestered by the same mechanisms de- organics As and Hg has been demonstrated for
scribed above, involving GSH-S-transferases, microorganisms, but these elements do not ap-
malonyl- and glucosyltransferases, and ATP- pear to be volatilized to signicant levels by
dependent conjugate-transport pumps (21). (nontransgenic) plants (105).
These degradation products of pollutants that Many VOCs can be volatilized passively by
accumulate in vacuoles or apoplast of plant tis- plants. Volatile pollutants with a Henrys law
sues are called bound residues (21). Atrazine and constant Hi >106 that are mobile in both air
TNT are examples of organic pollutants that and water can move readily from the soil via the
are partially degraded followed by storage of transpiration stream into the atmosphere (18).
the degradation products as bound residues (14, In this way, plants act like a wick for VOCs
22). For TCE, different results were obtained to facilitate their diffusion from soil. Examples
in different studies: Overall, TCE appears to of organic pollutants that can be volatilized by
be in part volatilized by the plant, part is stored plants are the chlorinated solvent TCE and the
as bound residue, and part may be completely fuel additive MTBE (26, 90).
degraded (111). Phytoremediation of TCE is Because volatilization completely removes
a much-studied process, and the remaining un- the pollutant from the site as a gas, with-
certainty about its fate illustrates that still much out need for plant harvesting and disposal,
remains to be learned about the metabolic fate this is an attractive technology. In the case
of organics in plants. Better knowledge in this of Se, the volatile form was also reported to
respect would be benecial not only for further be 23 orders of magnitude less toxic than
improvement of phytoremediation efciency, the inorganic Se forms (119). Volatilization
but also for better estimating the potential risks may be promoted in several ways. Although
involved. volatilization of VOCs is passive, the pro-
Phytodegradation of organic pollutants may cess may be maximized by using phreatophyte
be optimized by selecting or engineering plant species with high transpiration rates and by
species with higher activities of the enzymes promoting transpiration (preventing stomatal
thought to be involved and rate-limiting. There closure through sufcient irrigation). For Se,
are some examples of promising transgenic ap- enzymes of the S assimilation pathway medi-
proaches. The expression in plants of bacterial ate Se volatilization, and overexpression of one
enzymes involved in reductive transformation of these, cystathionine- -synthase promotes Se
of TNT (tetranitrate reductase or nitroreduc- volatilization (124). In another approach, the
tase) resulted in enhanced plant tolerance and enzyme SeCys methyltransferase from a Se
degradation of TNT (45, 51). Also, the consti- hyperaccumulator species was expressed in a
tutive expression of a mammalian cytochrome nonaccumulator, also signicantly enhancing
P450 in tobacco resulted in an up to 640-fold Se volatilization (71). Volatilization of mercury
higher ability to metabolize TCE (33). by plants was achieved by introducing a bacte-
rial mercury reductase (MerA). The resulting
plants volatilized elemental mercury and were
Volatilization signicantly more Hg-tolerant (105).
Phytovolatilization is the release of pollutants If a toxic volatile pollutant is emitted by
from the plant to the atmosphere as a gas. plants during phytoremediation, the fate of the
Inorganic Se can be volatilized by plants and gas in the atmosphere should be determined as
microorganisms. Volatilization of Se involves part of risk assessment. Such a study was done
assimilation of inorganic Se into the organic for volatile Se and Hg, and the pollutant was re-
selenoaminoacids selenocysteine (SeCys) and portedly dispersed and diluted to such an extent
selenomethionine (SeMet). The latter can be that it did not pose a threat (74, 85).

[Link] Phytoremediation 29
AR242-PP56-02 ARI 1 April 2005 13:36

NEW DEVELOPMENTS IN This has also resulted in practical phytoreme-

PHYTOREMEDIATION diation resources, such as online databases of
plant species that may be useful for cleanup of
In the past 10 years phytoremediation has
different types of pollutants (84) (PHYTOPET
gained acceptance as a technology and has been
lists species particularly useful for cleanup of
acknowledged as an area of research. There
petroleum hydrocarbons and PHYTOREM
has already been a substantial increase in our
lists plants that are recommended for metals
knowledge of the mechanisms that underlie the
and metalloids). The U.S. Environmental Pro-
uptake, transport, and detoxication of pollu-
tection Agency also maintains a phytoremedi-
tants by plants and their associated microbes.
ation Web site ([Link] with a
Still, large gaps in our knowledge await further
wealth of information for researchers and the
research, as indicated above. Phytoremediation
general public (e.g., citizens guides, phytore-
efciency is still limited by a lack of knowl-
mediation resource guide) (37, 38).
edge of many basic plant processes and plant-
Future eld phytoremediation projects
microbe interactions. There is also a need for
should benet from (more) collaboration be-
more phytoremediation eld studies to demon-
tween research groups and industry so that
strate the effectiveness of the technology and
they can be designed to address hypotheses and
increase its acceptance.
gain scientic knowledge in addition to meeting
Continued phytoremediation research
cleanup standards. Future eld phytoremedia-
should benet from a (more) multidisciplinary
tion projects will also benet from coordinated
approach, involving teams with expertise at all
experimental design across projects so that re-
organization levels, to study the remediation of
sults can be better compared.
pollutants from the molecule to the ecosystem.
An interesting development in phytoreme-
Phytoremediation research at universities is
diation is its integration with landscape ar-
generally carried out by scientists with exper-
chitecture. Remediation of urban sites (parks,
tise at a certain organizational level (e.g., plant
nature areas) may be combined with an attrac-
molecular biology, plant biochemistry, plant
tive design so that the area may be used by the
physiology, ecology, or microbiology) and of a
public during and after the remediation process
certain subset of pollutants (e.g., heavy metals,
while minimizing risk (66). Other sites that are
herbicides, TNT, or PAHs). Because research
phytoremediated may be turned into wildlife
on phytoremediation of organics and inorgan-
sanctuaries, like the Rocky Mountain Arse-
ics requires different expertise they are carried
nal in Denver, once one of the most polluted
out in different research communities, with
sites in the United States ([Link]
more engineers studying organics and more bi-
[Link]/).
ologists studying inorganics. These researchers
Another new development in phytoremedi-
do not interact optimally, in part because of a
ation is the use of transgenic plants. Knowledge
lack of phytoremediation conferences and sci-
gained from plant molecular studies in the past
entic journals that cover inorganics and organ-
10 years has led to the development of some
ics equally. Because 64% of polluted sites con-
promising transgenics that show higher toler-
tain mixtures of organics and inorganics (36),
ance, accumulation, and/or degradation capac-
phytoremediation would benet from more
ity for various pollutants, as described above. So
collaborative studies by teams of researchers
far, these transgenics have mainly been tested
from different backgrounds, to combine ex-
in laboratory studies using articially contami-
pertise in phytoremediation of both types of
nated medium rather than soils from the eld,
pollution and at multiple organization levels.
let alone eld studies. However, this is start-
Despite the remaining gaps in our knowl-
ing to change. One eld phytoremediation
edge, research has yielded much useful knowl-
study using transgenic Indian mustard plants
edge for phytoremediation, as described above.
that overexpress enzymes involved in sulfate/

30 Pilon-Smits
AR242-PP56-02 ARI 1 April 2005 13:36

selenate reduction and in accumulation of GSH shoot-specic expression of another) or that ex-
was just completed (96, 134, 135). Three types press a transgene only under certain environ-
of transgenic Indian mustard plants that over- mental conditions (31). Also, genetic engineer-
express enzymes involved in sulfate/selenate re- ing of the chloroplast genome offers a novel
duction and in accumulation of GSH showed way to obtain high expression without the risk
enhanced Se accumulation in the eld when of spreading the transgene via pollen (106). In
grown on soil polluted with Se, B, and other another totally new approach, it was shown to
salts (G. Banuelos, N. Terry, D. LeDuc, E. be possible to genetically manipulate an en-
Pilon-Smits & C. Mackey, unpublished re- dophytic microorganism, leading to enhanced
sults). Earlier, these same transgenics showed toluene degradation (10).
enhanced capacity to accumulate Se and heavy As transgenics are being tested in the eld
metals (Cd, Zn) from polluted soil from the eld and the associated risks assessed, their use may
in greenhouse experiments (12, 125). Another become more accepted and less regulated, as
eld experiment testing Hg volatilizing (MerA) has been the case for transgenic crops. Also,
poplar trees is presently underway (D. Glass, as more information becomes available about
personal communication). the movement of pollutants in ecosystems and
In the coming years, mining of the genomic the associated risks, the rules for cleanup tar-
sequences from Arabidopsis thaliana and rice and gets may be adjusted depending on future use
availability of new genomic technologies should of the site, bioavailability of the pollutant, and
lead to the identication of novel genes impor- form of the pollutant. Because phytoremedia-
tant for pollutant remediation, including regu- tion only remediates the bioavailable fraction
latory networks (e.g., transcription factors) and of the pollution, stringent cleanup targets limit
tissue-specic transporters. The expression of the applicability of this technology. If targets
these genes may then be manipulated in high- can be adjusted to focus on the bioavailable
biomass species for use in phytoremediation. (i.e., toxic) fraction of the pollutant, phytore-
Other new developments in plant genetic en- mediation could become more widely applica-
gineering are tailored transgenics that overex- ble. This would reduce cleanup costs and enable
press different enzymes in different plant parts the cleanup of more sites with the limited funds
(e.g., root-specic expression of one gene and available.

SUMMARY POINTS
1. Plants and their associated microbes can remediate pollutants via stabilization, degrada-
tion in the rhizosphere, degradation in the plant, accumulation in harvestable tissues, or
volatilization.
2. Phytoremediation offers a cost-effective and environment-friendly alternative or com-
plementary technology for conventional remediation methods such as soil incineration
or excavation and pump-and-treat systems.
3. Although phytoremediation works effectively for a wide range of organic and inorganic
pollutants, the underlying biological processes are still largely unknown in many cases.
Some important processes that require further study are plant-microbe interactions,
plant degradation mechanisms for organics, and plant transport and chelation mecha-
nisms for inorganics.
4. New knowledge and plant material obtained from research is being implemented for
phytoremediation in the eld. The rst eld tests with transgenic plants are showing

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AR242-PP56-02 ARI 1 April 2005 13:36

promising results. As more results demonstrating the effectiveness of phytoremediation

become available its use may continue to grow, reducing cleanup costs and enabling the
cleanup of more sites with the limited funds available.

ACKNOWLEDGMENTS
The authors research is supported by National Science Foundation Grant MCB9982432 and
U.S. Department of Agriculture NRI grant #2003-35318-13758.

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