Bioresource Technology 77 (2001) 229±236

Review paper

Phytoextraction: a cost-e€ective plant-based technology for the

removal of metals from the environment
Carlos Garbisu a,*, Itziar Alkorta b
a
Departamento de Agrosistemas y Producci on Animal, NEIKER, Instituto Vasco de Investigaci on y Desarrollo Agrario, Berreaga 1,
E-48160 Derio, Spain
b
Departamento de Bioquõmica y Biologõa Molecular, Facultad de Ciencias, Universidad del Paõs Vasco, Apartado 644, E-48080 Bilbao, Spain
Accepted 31 July 2000

Abstract
Phytoremediation is an emerging technology that uses plants to clean up pollutants (metals and organics) from the environment.
Within this ®eld of phytoremediation, the utilization of plants to transport and concentrate metals from the soil into the harvestable
parts of roots and above-ground shoots is usually called phytoextraction. Most traditional remediation methods do not provide
acceptable solutions for the removal of metals from soils. By contrast, phytoextraction of metals is a cost-e€ective approach that
uses metal-accumulating plants to clean up these soils. Subsequently, the harvestable parts, rich in accumulated metals, can be easily
and safely processed by drying, ashing or composting. Some extracted metals can also be reclaimed from the ash, generating re-
cycling revenues. Phytoextraction appears a very promising technology for the removal of metal pollutants from the environment
and may be, at present, approaching comercialization. Ó 2001 Elsevier Science Ltd. All rights reserved.

Keywords: Phytoextraction; Metal; Phytoremediation; Soil pollution

1. Introduction: soil pollution and heavy metals
Soil pollution, a very important environmental
problem, has been attracting considerable public atten-
tion over the last decades. As a matter of fact, increas-
ingly widespread pollution has caused vast areas of land
to become non-arable and hazardous for both wildlife
and human populations. Nowadays, more and more
people consider that the magnitude of the pollution
problem in our soils calls for immediate action. Unfor-
tunately, the enormous costs associated with the re-
moval of pollutants from soils by means of traditional
physicochemical methods have been encouraging
companies to ignore the problem. In fact, the current
state-of-the-art technology for the remediation of metal-
polluted soils is the excavation and burial of the soil at a
hazardous waste site at an average cost of $1 000 000 per
acre (Raskin et al., 1997). Other common approaches
used to treat metal-polluted soils are ®xation (chemical
processing of the soil to immobilize the metals) and
leaching (using acid solutions or proprietary leachants
*
Corresponding author. Fax: +349-4452-2335.
E-mail address: cgarbisu@neiker.net (C. Garbisu).
to desorb and leach metals from soil followed by the
return of clean soil residue to the site) (Salt et al., 1995).
Apart from minimizing the impact of future incidents by
means of controlling pollution input, it is imperative to
deploy innovative technologies which could economi-
cally remediate polluted soils (Garbisu and Alkorta,
1997).
In contrast to many organic pollutants, which are
anthropogenic and often degraded in the soil, metals
occur naturally and are conserved (Wade et al., 1993).
Due to their immutable nature, heavy metals are a
group of pollutants of much concern. The danger of
heavy metals is aggravated by their almost inde®nite
persistence in the environment. Although some metals
are essential for life (i.e., they provide essential cofactors
for metalloproteins and enzymes), at high concentra-
tions they can act in a deleterious manner by blocking
essential functional groups, displacing other metal ions,
or modifying the active conformation of biological
molecules (Collins and Stotzky, 1989). In addition, they
are toxic for both higher organisms and microorgan-
isms. In fact, many metals a€ect directly various physi-
ological and biochemical processes causing reduction in
growth, inhibition of photosynthesis and respiration,
and degeneration of main cell organelles (Vangronsveld

0960-8524/01/$ - see front matter Ó 2001 Elsevier Science Ltd. All rights reserved.
PII: S 0 9 6 0 - 8 5 2 4 ( 0 0 ) 0 0 1 0 8 - 5
230 C. Garbisu, I. Alkorta / Bioresource Technology 77 (2001) 229±236
and Clijsters, 1994). Some metals are accumulated in
roots (especially Pb), probably due to some physiologi-
cal barriers against metal transport to aerial parts, while
others are easily transported in plants, for example, Cd.
Heavy metals cannot be destroyed biologically (no
``degradation'', change in the nuclear structure of the
element, occurs) but are only transformed from one
oxidation state or organic complex to another. As a
consequence of the alteration of its oxidation state, the
metal may become either: (i) more water soluble and is
removed by leaching, (ii) inherently less toxic, (iii) less
water soluble so that it precipitates and then becomes
less bioavailable or removed from the contaminated site,
or (iv) volatilized and removed from the polluted area
(Garbisu and Alkorta, 1997). Heavy metals are present
in soil as natural components or as a result of human
activity. The primary sources of metal pollution are the
burning of fossil fuels, mining and smelting of metal-
liferous ores, downwash from power lines, municipal
wastes, fertilizers, pesticides and sewage.
2. Microbial remediation of metal-polluted soils
Microorganisms can detoxify metals by valence
transformation, extracellular chemical precipitation, or
volatilization. In fact, some microorganisms can enzy-
matically reduce a variety of metals in metabolic pro-
cesses that are not related to metal assimilation (Lovley,
1993): some bacteria obtain energy for growth by cou-
pling the oxidation of simple organic acids and alcohols,
hydrogen, or aromatic compounds, to the reduction of
Fe(III) or Mn(IV). Bacteria that use U(VI) as a terminal
electron acceptor may be useful for removing uranium
from contaminated sites. The reduction of the toxic
selenate and selenite to the insoluble and much less toxic
elemental selenium may be exploited to enhance re-
moval of these anions from contaminated sites (Combs
et al., 1996; Garbisu et al., 1995a, 1995b; 1997a, Garbisu
and Alkorta, 1997). The more toxic form of chromium,
Cr(VI), can also be detoxi®ed by bacterially mediated
reduction and the enzymatic mechanism responsible for
the reduction of Cr(VI) to Cr(III) is currently being
studied and may ultimately lead to a commercial bio-
remediation process (Garbisu et al., 1997b,c, 1998;
Ishibashi et al., 1990). Another natural reduction pro-
cess now being developed for commercial applications is
the transformation of mercuric ion, Hg(II), to volatile
metallic mercury, Hg(0). Microorganisms can also en-
zymatically reduce other metals such as technetium,
vanadium, molybdenum, gold, silver and copper, but
reduction of these metals has not been studied exten-
sively (Lovley, 1993).
Although it is true that microorganisms that use
metals as terminal electron acceptors or reduce them as
a detoxi®cation mechanism can be of use for the re-
moval of these pollutants from the environment (Gar-
bisu and Alkorta, 1997), it is certainly not less true that
when considering the remediation of a metal-polluted
soil, metal-accumulating plants o€er numerous advan-
tages over microbial processes since plants can actually
extract metals from the polluted soils, theoretically
rendering them clean (metal-free soils). In fact, although
a wide variety of bacterial, fungal, algal and plant sys-
tems are capable of concentrating toxic metals from
their surroundings, so far no cost-e€ective way exists to
retrieve small organisms from the soil (Ow, 1996).
Therefore, and in relation to the bioremediation of
heavy metals, microorganisms have been mostly used to
treat industrial waste streams, with the organisms either
immobilized onto di€erent support matrixes or in a free-
living state, enclosed in treatment tanks or other kinds
of reactor vessels. Subsequently, the metal-loaded bio-
mass can be either disposed of appropriately or, de-
pending on their concentrations, treated to recover the
metals. In the environment, as is the case for the in situ
bioremediation systems, bacteria are not e€ective as a
permanent, large-scale solution to heavy metal-polluted
areas, since this implies the ultimate removal of the
contaminated biomass from the site. As a consequence,
application of microbial bioremediation to the in situ
removal of heavy metals from polluted soils is mainly
limited to metal immobilization by precipitation or re-
duction (Summers, 1992).
Phytoremediation is also preferable to currently used
approaches for treating soils such as land®lling, ®xation,
leaching, etc., since it reclaims soil at the site, recycling it
in a biologically safe state rather than permanently
disposing of it by removal to a storage site (Salt et al.,
1995).
3. Phytoremediation
3.1. General aspects of phytoremediation
Phytoremediation, de®ned as the use of green plants
to remove pollutants from the environment or to render
them harmless (Cunningham and Berti, 1993; Raskin
et al., 1994), is being considered as a new highly prom-
ising technology for the remediation of polluted sites.
Phytoremediation is often also referred as botanical-
bioremediation or green remediation (Chaney et al.,
1997). This technology can be applied to both organic
and inorganic pollutants present in soil (solid substrate),
water (liquid substrate) or the air (Salt et al., 1998). In
this respect, plants can be compared to solar driven
pumps which can extract and concentrate certain ele-
ments from their environment (Salt et al., 1995). How-
ever, the ability to accumulate heavy metals varies
signi®cantly between species and between cultivars
within a species.
 C. Garbisu, I. Alkorta / Bioresource Technology 77 (2001) 229±236
Following harvest of pollutant-enriched plants, the
weight and volume of contaminated material can be
further reduced by ashing or composting. Metal-en-
riched plants can be disposed of as hazardous material
or, if economically feasible, used for metal recovery (Salt
et al., 1998).
Most existing remediation physicochemical technol-
ogies are meant primarily for intensive in situ or ex situ
treatment of relatively highly polluted sites, and thus are
not very suitable for the remediation of vast, di€usely
polluted areas where pollutants occur only at relatively
low concentrations and super®cially (Rulkens et al.,
1998). In this context, phytoremediation appears as a
very valid option since it is best suited for the remedi-
ation of these di€usely polluted areas and at much lower
costs than other methods. While the most heavily con-
taminated soils do not support plant growth, sites with
light to moderate toxic metal contamination can be
remediated by growing metal-accumulating plants
(Kumar et al., 1995). Examples of pollutants that could
possibly be removed by phytoremediation are heavy
metals, 2,4,6-trinitrotoluene, trichloroethylene, benzene,
toluene, ethylbenzene, and xylene (Rulkens et al., 1998).
More and more often, and provided that it will result
in lower costs and that the risks posed to human pop-
ulations and ecosystems are acceptable, the fact that the
remediation procedure may take rather long, as is the
case with phytoremediation methods, is not considered a
problem (Rulkens et al., 1998).
Interestingly enough, in the last few years, the pos-
sibility of planting metal hyperaccumulator crops over a
low-grade ore body or mineralized soil, and then har-
vesting and incinerating the biomass to produce a
commercial ``bio-ore'' has been proposed (Brooks et al.,
1998). This ``phytomining'' o€ers the possibility of ex-
ploiting ore bodies that are otherwise uneconomic to
mine and its e€ect on the environment is minimal when
compared with the erosion caused by opencast mining
(Brooks et al., 1998).
As a result of their association with speci®c ore de-
posits, many metallophyte plants are used in prospecting
for mineral deposits (Brooks, 1983).
3.2. Categories of phytoremediation
These include (Chaney et al., 1997; Salt et al., 1995,
1998; Raskin et al., 1997).
Phytoextraction: the use of plants to remove con-
taminants from soils. Pollutant-accumulating plants are
utilized to transport and concentrate contaminants
(metals or organics) from the soil into the above-ground
shoots; the term is mostly used to refer to metal removal
from soils. In some cases, roots can be harvested as well
(Kumar et al., 1995).
Phyto®ltration: the use of plant roots (rhizo®ltration)
or seedlings (blasto®ltration) to absorb or adsorb pol-
231
lutants, mainly metals, from water and aqueous-waste
streams. Plant roots or seedlings grown in aerated water
absorb, precipitate and concentrate toxic metals from
polluted e‚uents.
Phytostabilization: the use of plants to reduce the
bioavailability of pollutants in the environment. Plants
stabilize pollutants in soils, thus rendering them harm-
less and reducing the risk of further environmental de-
gradation by leaching of pollutants into the ground
water or by airborne spread.
Phytovolatilization: the use of plants to volatilize
pollutants. Plants extract volatile pollutants (e.g., sele-
nium, mercury) from soil and volatilize them from the
foliage.
Phytodegradation: the use of plants and associated
microorganisms (plant-assisted bioremediation) to de-
grade organic pollutants. Plant roots in conjunction
with their rhizospheric microorganisms are utilized to
remediate soils contaminated with organics; the air pu-
rifying uses of some plants.
Some authors (Stomp et al., 1994) also distinguish
between indirect and direct phytoremediation. In the case
of indirect phytoremediation, plants participate in the
detoxi®cation of pollutants via their support of symbi-
otic, root-associated microorganisms that actually ac-
complish contaminant detoxi®cation (this matches the
previously named plant-assisted bioremediation). On the
other hand, plants could participate directly through
contaminant uptake and subsequent contaminant im-
mobilization or degradation within the plant.
3.3. Use of trees
According to some authors (Stomp et al., 1994), trees
potentially are the lowest-cost plant type to use for
phytoremediation. A number of tree species can grow on
land of marginal quality. This allows establishment of
trees on sites with low fertility and poor soil structure,
keeping costs low for plant establishment. Besides, trees
have the most massive root systems of all plants, which
penetrate the soil for several meters, farther than most
herbaceous plants (Stomp et al., 1994). In some tree
species above-ground biomass can be harvested, and
trees will resprout without disturbance of the site. This
attribute would be valuable if periodic removal of pol-
lutants sequestered in plant tissue were desirable as in
the case of heavy metals bound to wood (Stomp et al.,
1994).
3.4. Conclusions
With a sound footing in basic research and a new
regulatory environment supportive of innovative tech-
nologies, phytoremediation is increasingly being viewed
as a cost-e€ective and user-friendly alternative to tradi-
tional methods of environmental cleanup (Boyajian and
232 C. Garbisu, I. Alkorta / Bioresource Technology 77 (2001) 229±236
Carreira, 1997). Optimizing agronomic practices, such
as irrigation, fertilization, planting and harvest time and
the timing of amendment application, will increase the
eciency of the phytoremediation processes (Ensley
et al., 1997).
Phytoremediation is still a new ®eld which holds great
potential and in order to develop this potential requires
a multidisciplinary approach, spanning ®elds as diverse
as plant biology, agricultural engineering, agronomy,
soil science, microbiology and genetic engineering.
4. Phytoextraction of metals
The term ``phytoextraction'' mainly concerns the re-
moval of heavy metals or radionuclides from soil by
means of the uptake capabilities of plants. Plants can
accumulate heavy metals essential for growth and de-
velopment such as Fe, Mn, Zn, Cu, Mg, Mo, and pos-
sibly Ni. In addition, some of them have the capacity to
accumulate heavy metals with no known biological
functions, such as Cd, Cr, Pb, Co, Ag, Se and Hg (Baker
and Brooks, 1989; Raskin et al., 1994). For the metab-
olism of metals, plants require a balance between the
uptake of essential metal ions to maintain growth and
development and the ability to protect sensitive cellular
activity and structures from excessive levels of essential
and non-essential metals. Resistance of plants to heavy
metal ions can be achieved by an avoidance mechanism,
which includes mainly the immobilization of metal in
root and in cell walls. Tolerance to heavy metals is based
on the sequestration of heavy metal ions in vacuoles, on
binding them by appropriate ligands like organic acids,
proteins and peptides and on the presence of enzymes
that can function at high levels of metallic ions (Har-
borne, 1989; Robinson et al., 1994).
The phytoextraction of heavy metals represents one
of the largest economic opportunities for phytoremedi-
ation because of the size and scope of environmental
problems associated with metal-contaminated soils and
the competitive advantage o€ered by a plant-based
remediation technology (Raskin et al., 1997).
The ideal plant to be used in phytoextraction should
have the following characteristics:
· be tolerant to high levels of the metal;
· accumulate high levels of the metal in its harvestable
parts;
· have a rapid growth rate;
· have the potential to produce a high biomass in the
®eld;
· have a profuse root system.
The idea of using plants to remove metals from soils
came from the discovery of di€erent wild plants, often
endemic to naturally mineralized soils, that accumulate
high concentrations of metals in their foliage (Brooks
et al., 1979; Baker and Brooks, 1989; Raskin et al., 1997).
Curiously enough, the capacity of plants to concen-
trate metals has usually been considered a detrimental
trait since some plants are directly or indirectly re-
sponsible for a proportion of the dietary uptake of toxic
heavy metals by humans (Brown et al., 1994; Cunning-
ham et al., 1995). Dietary intake of heavy metals
through consumption of crop plants can have long-term
e€ects on human health (Ow, 1996). Naturally occurring
plants called ``metal hyperaccumulators'' can accumu-
late 10±500 times higher levels of elements than crops
(Chaney et al., 1997). According to Raskin et al. (1997),
the degree of accumulation of metals such as Ni, Zn,
and possibly Cu, observed in hyperaccumulators, often
reaches 1±5% of their dry weight (an order of magnitude
higher than those concentrations found in non-accu-
mulating plants growing nearby). Unfortunately, most
hyperaccumulators are relatively small in size, have slow
growth rates and we lack the technology for their large-
scale cultivation (Salt et al., 1995; Raskin et al., 1997).
The annual yields in biomass of hyperaccumulators are
generally one to two orders of magnitude lower than
those of robust crop plants (Ow, 1996). Therefore, a lot
of research emphasis has been placed on the evaluation
of the metal-accumulating capacity of high biomass
plants that can be easily cultivated using established
agronomic practices. In this respect, many authors have
studied the metal-uptake capabilities of the cultivated
Brassica (mustard) species because of their relation to
wild metal-accumulating mustards (Kumar et al., 1995).
Either the existing hyperaccumulator plants must be
bred for increased growth and biomass, or hyperaccu-
mulation traits must be engineered into fast growing,
high biomass plants (Ow, 1996). However, according to
some authors (Chaney et al., 1997), natural metal hy-
peraccumulator phenotype appears to be much more
important than high yield ability when using plants to
remediate metal-contaminated soils.
The prevention of herbivory and disease is thought to
be the main function of this unique phenomenon of
hyperaccumulation (Baker and Brooks, 1989; Ernst
et al., 1990; Boyd and Martens, 1994, 1995; Boyd et al.,
1994).
As pointed out in the excellent review by Salt et al.
(1998) there are, at present, two strategies of phytoex-
traction: (1) chelate-assisted or induced phytoextraction
and (2) continuous phytoextraction.
4.1. Chelate-assisted or induced phytoextraction
Chelating agents have been used as soil extractants, a
source for micronutrient fertilizers, and to maintain
solubility of micronutrients in hydroponic solutions
(Salt et al., 1995). As pointed out by its name, this
strategy of phytoextraction is based on the fact that the
application of metal chelates to the soil signi®cantly
enhances metal accumulation by plants. This is due to
 C. Garbisu, I. Alkorta / Bioresource Technology 77 (2001) 229±236
the fact that under many circumstances, in the soil and
depending on the metal itself, it is common to ®nd cases
of low bioavailability, preventing the remediation pro-
cess. Due to this phenomenon of the low bioavailability
of metals in soils, it is common to observe that shoot
metal accumulation in hydroponically cultivated plants
greatly exceeds the metal accumulation measured in soil-
grown plants. For instance, lead (Pb) is usually ex-
tremely insoluble (i.e., not available for plant uptake) in
the normal range of soil pH (Raskin et al., 1997). Thus,
vegetation growing in heavily contaminated areas often
has less than 50 mg/g Pb in shoots (Cunningham et al.,
1995). Corn (Zea mays) and, to a lesser extent, ragweed
(Ambrosia artemisii®lia) have been identi®ed as good
accumulators of Pb (Huang and Cunningham, 1996).
But even plants, such as Brassica juncea, that have a
genetic capacity to accumulate Pb, will not contain
much Pb in roots or shoots if cultivated in Pb-contam-
inated soil (Raskin et al., 1997). A large proportion of
many metals remains sorbed to solid soil constituents.
The formation of metal±chelate complexes prevents
precipitation and sorption of the metals thereby main-
taining their availability for plant uptake (Salt et al.,
1995). The addition of chelates to the soil can also bring
metals into solution through desorption of sorbed spe-
cies, dissolution of Fe and Mn oxides, and dissolution of
precipitated compounds (Norwell, 1984).
Fortunately, the discovery that the application of
certain chelates to the soil increases the translocation of
heavy metals from soil into the shoots has opened a wide
new range of possibilities for this ®eld of metal phy-
toextraction (Blaylock et al., 1997). Ethylenediamine-
tetraacetic acid (EDTA), when applied to established
plants several days before harvest, has proven to be very
e€ective in facilitating the plant uptake of Cd, Cu, Ni,
Pb and Zn (Raskin et al., 1997). There are no reliable
reports on the natural accumulation of the most envi-
ronmentally important metallic pollutants (i.e., lead,
cadmium, arsenic, radionuclides such as U, Cs, Sr) (Salt
et al. 1998).
However, it has been described that high biomass
crop plants such as Indian mustard, corn and sun¯ower
could accumulate signi®cant amounts of Pb when in-
duced through the addition of metal chelates (Huang
and Cunningham, 1996; Blaylock et al., 1997). Blaylock
et al. (1997) demonstrated the simultaneous accumula-
tion of several metals (Pb, Cd, Cu, Ni, Zn) by Indian
mustard plants after applying metal chelates. According
to Blaylock et al. (1997) and Huang and Cunningham
(1996), plants can remove between 180 and 530 kg/ha of
Pb per year, making remediation of sites contaminated
with up to 2500 mg/kg Pb possible in under 10 years
(Salt et al., 1998). Metal accumulation eciency appears
to be directly related to the anity of the applied chelate
for the metal (Salt et al., 1998). Therefore, it can be
concluded that, for ecient phytoextraction to occur,
233
synthetic chelates having a high anity for the metal of
interest should be used: EDTA for lead, EGTA for
cadmium (Blaylock et al., 1997), possibly citrate for
uranium, etc. The mechanisms involved in metal-chelate
induced plant uptake and translocation of metals are
not well understood (Salt et al., 1998). Nonetheless, it is
likely that Pb is transported within plants as a Pb±
EDTA complex.
Apart from the addition of synthetic chelates, plants
secrete to the rhizosphere natural metal-chelating mol-
ecules to mobilize soil-bound metals. So far, only
phytosiderophores, iron-chelating compounds have
been studied in detail. Some of these phytosiderophores
include mugeneic and deoxymugeneic acids from barley
and corn, and avenic acid from oats (Welch et al., 1993).
There is also the possibility that metal-chelating pro-
teins, perhaps related to metallothioneins or phyto-
chelatins, may act as phytosiderophores (Raskin et al.,
1997).
Finally, plants roots increase metal bioavailability by
extruding protons to acidify the soil and mobilize the
metals. A similar mechanism has been observed for Fe
mobilization in some Fe-de®cient dicotyledonous plants
(Crowley et al., 1991). In fact, numerous studies have
shown that lowering the pH of a soil decreases the ad-
sorption of heavy metals and thus increases their con-
centration in the soil solution (Harter, 1983; Salt et al.
1995). It may be possible to increase metal availability
(and hence plant uptake) by means of maintaining a
moderately acid pH in the soil through the use of
ammonium containing fertilizers or soil acidi®ers (Salt
et al., 1995).
Roots can also reduce soil-bound metal ions by spe-
ci®c plasma-membrane bound metal reductases. For
instance, pea plants de®cient in Fe or Cu have an in-
creased ability to reduce Fe3‡ and Cu2‡ , which is cou-
pled with an increased uptake of Cu, Mn, Fe and Mg
(Welch et al., 1993).
4.2. Continuous phytoextraction
Continuous phytoextraction depends on the natural
ability of some plants to accumulate, translocate and
resist high amounts of metals over the complete growth
cycle. In this context, hyperaccumulators are the most
suitable plants for the phytoextraction of metal-polluted
soils. Unfortunately, as mentioned above, hyperaccu-
mulators for the most environmentally important me-
tallic pollutants (Pb, Cd, As, U) have not been reliably
described (Salt et al., 1998). Nonetheless, hyperaccu-
mulators can naturally accumulate more than 1% of
shoot dry biomass as Zn, Ni, or Se (Ba~ nuelos and
Meeks, 1990; Bauman, 1885; Brooks et al., 1979; Byers,
1935, 1936; Chaney et al., 1995; Reeves and Brooks,
1983; Salt et al., 1995, 1998). The existence of hyperac-
cumulators for metals other than Ni, Zn and Se is not
234 C. Garbisu, I. Alkorta / Bioresource Technology 77 (2001) 229±236
very clear and needs further substantiation (Salt et al.,
1998). However, it is important to emphasize that even
the best metal accumulators, such as Thlaspi caerules-
cens (Brassicaceae) (Baker et al., 1994), take a relatively
long time of continuous cultivation (13±14 years) to
clean a site of Ni or Zn (Salt et al., 1995).
Most metal-accumulating plant species known today
were discovered growing on soils containing high levels
of heavy metals. These plants are often endemic to these
types of soils, suggesting that metal accumulation is
associated with heavy metal resistance (Baker and
Brooks, 1989). The majority of hyperaccumulating
species discovered so far are restricted to tropical areas
(Baker and Brooks, 1989; Baker et al., 1993; Brooks
et al., 1993). The ®rst hyperaccumulators characterized
were members of the Brassicaceae and Fabaceae fami-
lies. The number of metal-accumulating taxa identi®ed
to date has been reported to be 397 (Salt et al., 1998).
Therefore, there is a continuous search for novel phy-
toextracting plants adapted to particular ecosystems and
climates. In general, and as mentioned above, some
studies (Salt et al., 1995) have demonstrated that the
ability to accumulate heavy metals varies greatly be-
tween species and between cultivars within a species.
Particular emphasis has been placed on the evaluation
of shoot metal-accumulation capacity of the cultivated
Brassica (mustard) species because of their relation to
wild metal-accumulating mustards (Kumar et al., 1995).
Although the largest numbers of temperate-climate hy-
peraccumulating species belong to the Brassicaceae
(Baker and Brooks, 1989), in the tropics the Euphorbi-
aceae is the best represented group (Ensley et al., 1997).
Several examples of ®eld trials for the continuous
phytoextraction of metals have been reported: Thlaspi
caerulescens and Silene vulgaris for Cd and Zn, respec-
tively (Brown et al., 1995); Brassica oleracea, Raphanus
sativus, Thlaspi caerulescens, Alyssum lesbiacum, Alys-
sum murale and Arabidopsis thaliana for Zn, Cd, Ni, Cu,
Pb and Cr, respectively (Baker et al., 1991).
4.2.1. Microbes associated with phytoextraction
Plant-assisted bioremediation has been mainly con-
cerned with the degradation of organic pollutants and
the use of microorganisms to improve the plant±metal
uptake from soils has hardly been investigated. Roots
can employ rhizospheric organisms (mycorrhizal fungi
or root-colonizing bacteria) to increase the bioavail-
ability of metals (Raskin et al., 1997). However, it is
believed that plant uptake of certain mineral nutrients
such as Fe and Mn may be facilitated by rhizospheric
microorganisms (Barber and Lee, 1974; Crowley et al.,
1991). Similar results may be found for non-essential
heavy metals. Several strains of Bacillus and Pseudo-
monas increased the total amount of Cd accumulated by
Brassica juncea seedlings (Salt et al., 1995). From these
studies, it can be concluded that by populating the
rhizosphere with selected microorganisms during the
phytoextraction, it should be possible to enhance uptake
of heavy metals from soils.
4.2.2. Metal reactions in plants
Once inside the plant, most metals are too insoluble
to move freely in the vascular system so they usually
form carbonate, sulfate or phosphate precipitates im-
mobilizing them in apoplastic (extracellular) and sym-
plastic (intracellular) compartments (Raskin et al.,
1997). Unless the metal ion is transported as a non-
cationic metal chelate, apoplastic transport is further
limited by the high cation-exchange capacity of cell walls
(Raskin et al., 1997). In addition, some metals may be
transported to the shoots complexed to organic acids,
mainly citrate (Baker and Brooks, 1989; Senden et al.,
1992).
4.2.3. Genetics of phytoextraction
One way to supplement the arsenal of plants available
for remedial actions is to utilize genetic engineering tools
to insert into plants those genes that will enable the
plant to metabolize a particular pollutant (Newman
et al., 1997). In fact, one of the goals of plant genetic
engineering is to enhance the ability of plants to me-
tabolize many of the compounds that are of environ-
mental concern. Some laboratories are using traditional
breeding techniques, others are creating protoplast-
fusion hybrids, and still others are looking at the direct
insertion of novel genes to enhance the metabolic
capabilities of a variety of plants (Newman et al., 1997).
Major long-term improvements in phytoextraction
should come when scientists isolate genes from various
plant, bacterial and animal sources that can enhance the
metal-accumulating potential of the plants in which
these genes are inserted. Actually, there is increased
support for the idea that genetically modi®ed plants can
play an important role in extracting heavy metals from
contaminated sources (Mo€at, 1995). Besides, if the high
yield and hyperaccumulation traits could be bred into a
single plant, the e€ectiveness of metal extraction could
increase by one or two orders of magnitude (Ow, 1996).
By inserting an altered mercuric ion reductase gene
(merA) into Arabidopsis thaliana, Rugh et al. (1996)
reported the production of a mercury-resistant trans-
genic plant that volatilized mercury into the atmosphere.
Results to date suggest that the cost of phytoremedia-
tion of mercury-contaminated soils will be one-tenth to
one-hundredth the cost of other traditional engineering
methods, including land®lling, thermal treatments, and
chemical extraction.
Finally, to point out that although phytoextraction is
still an active topic of research, small-scale ®eld trials
with wild metal accumulators collected from naturally
contaminated soils have demonstrated the feasibility of
the phytoextraction approach.
 C. Garbisu, I. Alkorta / Bioresource Technology 77 (2001) 229±236
Phytoextraction appears a very promising technology
for the removal of metal pollutants from the environ-
ment and may be, at present, approaching comercial-
ization.
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