Phyto Extraction

Review
Approaches for enhanced phytoextraction of heavy metals

Atul Bhargava
a,
*
, Francisco F. Carmona
b
, Meenakshi Bhargava
c,1
, Shilpi Srivastava
a
a
Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus, Gomti Nagar, Lucknow, UP 226010, India
b
Departamento de Agricultura del Desierto y Biotecnologa, Universidad de Arturo Prat, Avenida Arturo Prat 2120, Campus Huayquique, Iquique, Chile
c
Department of Chemistry, University of Allahabad, Allahabad, India
a r t i c l e i n f o
Article history:
Received 27 June 2011
Received in revised form
20 March 2012
Accepted 1 April 2012
Available online 27 April 2012
Keywords:
Phytoextraction
Heavy metals
Biochemical mechanisms
Cation transporters
Metallothioneins
Phytochelatins
Plant improvement
Transgenics
a b s t r a c t
The contamination of the environment with toxic metals has become a worldwide problem. Metal
toxicity affects crop yields, soil biomass and fertility. Soils polluted with heavy metals pose a serious
health hazard to humans as well as plants and animals, and often requires soil remediation practices.
Phytoextraction refers to the uptake of contaminants from soil or water by plant roots and their trans-
location to any harvestable plant part. Phytoextraction has the potential to remove contaminants and
promote long-term cleanup of soil or wastewater. The success of phytoextraction as a potential envi-
ronmental cleanup technology depends on factors like metal availability for uptake, as well as plants
ability to absorb and accumulate metals in aerial parts. Efforts are ongoing to understand the genetics
and biochemistry of metal uptake, transport and storage in hyperaccumulator plants so as to be able to
develop transgenic plants with improved phytoremediation capability. Many plant species are being
investigated to determine their usefulness for phytoextraction, especially high biomass crops. The
present review aims to give an updated version of information available with respect to metal tolerance
and accumulation mechanisms in plants, as well as on the environmental and genetic factors affecting
heavy metal uptake. The genetic tools of classical breeding and genetic engineering have opened the
door to creation of remediation cultivars. An overview is presented on the possible strategies for
developing novel genotypes with increased metal accumulation and tolerance to toxicity.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
The rising rate of human activities in the biosphere is posing
unprecedented threats that would ultimately lead to a disturbing
imbalance in the biosphere. Industrial activities such as chemical
works, service stations, metal fabrication shops, paper mills,
tanneries, textile plants, waste disposal sites and intensive agri-
culture are particularly guilty of polluting the environment
(Wong, 2003; Freitas et al., 2004). Heavy metal contamination of
soils has become a serious problem in areas of intense industry
and agriculture. A heavy metal is a member of an ill-dened
subset of elements that exhibit metallic properties, which mainly
includes the transition metals, some metalloids, lanthanides and
actinides (Babula et al., 2008). These are metallic chemical
elements that have a relatively high density and are toxic even at
low concentrations. The contamination of the environment with
heavy metals has become a worldwide problem that affects crop
yields, soil biomass and fertility, and leads to bioaccumulation of
metals in the food chain (Gratao et al., 2005; Rajkumar et al.,
2009). This is majorly due to pollution of agricultural soils by
increasing reliance on chemical fertilizers, which has imposed
a long-termrisk on environmental health (McLaughlin et al., 1999;
Wong et al., 2002). Industrialized countries have regulated the
emission of toxic substances, but in developing countries, rapid
industrial development and population explosion, coupled with
lack of pollution control has caused an enormous increase in
heavy metal contamination of agricultural soils (Ji et al., 2000).
Soils polluted with heavy metals pose a health hazard to humans
as well as plants and animals. Thus, heavy metals need to be
removed from the soil for agro-ecological sustainability and
human benet.
Various efcient soil cleanup techniques are available, but
most of them are costly, labour intensive and cause soil distur-
bances, and have thus found limited acceptability among the
communities. Conventional remediation methods usually involve
pneumatic fracturing, solidication/stabilization, vitrication,
excavation and removal of contaminated soil layer, physical
stabilization or washing of contaminated soils with strong acids or
heavy metal chelators (Steele and Pichtel, 1998; Khan et al., 2004;
* Corresponding author. Tel.: 91 522 2721931/32; fax: 91 522 2721934.
E-mail address: atul_238@rediffmail.com (A. Bhargava).
1
Present address: Delhi Public School, Indira Nagar, Lucknow, India.
Contents lists available at SciVerse ScienceDirect
Journal of Environmental Management
j ournal homepage: www. el sevi er. com/ l ocat e/ j envman
0301-4797/$ e see front matter 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jenvman.2012.04.002
Journal of Environmental Management 105 (2012) 103e120
Bhargava et al., 2012). Phytoremediation, using plants to cleanup
contaminated environment, is an idea that is attracting increasing
attention among scientists, remediation engineers, and environ-
mental professionals in government and industries. In situ
remediation using plants has the potential to be less expensive
than current technologies and simultaneously initiates both
detoxication of hazardous waste and site restoration. Phytor-
emediation technologies include phytoextraction, phytostabili-
zation, phytovolatilization, phytoltration and phytodegradation
(Fulekar et al., 2009; Marques et al., 2009). Phytoextraction
refers to the uptake of contaminants from soil or water by plant
roots and their translocation into the shoot, or any other
harvestable plant part, to remove contaminants and promote
long-term cleanup of soil or wastewater (Sas-Nowosielska et al.,
2008). In this approach, plants capable of accumulating heavy
metals are grown on contaminated sites and the metal-rich
aboveground biomass is harvested on maturity. As a result,
a fraction of the soil contaminant is removed. The success of
phytoextraction as a potential environmental cleanup technology
depends on factors like metal availability for uptake as well as
plant ability to absorb and accumulate metals in its aerial parts. As
per the economic feasibility, the harvested biomass is usually
incinerated or composted and rarely recycled for reuse (Prasad
and Freitas, 2003).
Plants ideal for phytoextraction should possess multiple traits
like ability to growoutside their area of collection, fast growth, high
biomass, easy harvesting and accumulation of a range of heavy
metals in their harvestable parts (Jabeen et al., 2009; Seth, 2011).
No plant is presently known that fulls all these criteria. However,
a rapidly growing non-accumulator plant could be modied and/or
engineered so that it achieves most of the above-mentioned attri-
butes. There has been signicant progress in determining the
biochemical and molecular basis for metal accumulation, which
provides us with a strong scientic basis to outline some strategies
for achieving this goal. This article rst aims to review studies that
are beginning to uncover the detailed mechanism behind phy-
toextraction of heavy metals in plants. The progress of research
focused on the unravelling of genetic and biochemical mechanisms
that confer the ability to accumulate heavy metals into plants is
described. Plant breeders and environmental researchers have long
strived to develop improved plant varieties which can be used for
effective phytoextraction. This aspect is been discussed in this
review, and the use of classical and biotechnological approaches
used for enhancing natural hyperaccumulation of heavy metals is
presented.
2. Plants as accumulators of heavy metals
The uptake of metals by plants is selective, with some being
preferentially acquired over others. Many heavy metals like nickel
(Ni), copper (Cu), manganese (Mn) and zinc (Zn) are essential
micronutrients and required by plants to grow and complete the
life cycle. According to Baker (1981), plants growing on metallif-
erous soils can be grouped into three categories: (i) Excluders e in
which the metal concentrations in the shoot are maintained up to
a critical value, at a low level across a wide range of soil concen-
tration. Excluders prevent uptake of toxic metals into root cells
(de Vos et al., 1991). Excluders can be used for stabilization of soil
and to avoid further spread of contamination due to erosion (Lasat,
2002). (ii) Accumulators e in which the metals are concentrated in
aboveground plant parts from low to high soil concentrations.
Accumulators do not prevent metals from entering the roots and
thus allow bioaccumulation of high concentration of metals.
(iii) Indicators e where internal concentration reects the external
levels (McGrath et al., 2002).
2.1. Hyperaccumulators
The discovery of hyperaccumulator plant species has revolu-
tionized phytomediation technology since these plants have an
innate capacity to absorb metal at levels 50e500 times greater than
average plants (Lasat, 2000). Hyperaccumulators are a subgroup of
accumulator species often endemic to naturally mineralized soils,
which accumulate high concentrations of metals in their foliage
(Baker and Brooks, 1989; Raskin et al., 1997). Metal hyper-
accumulators are naturally capable of accumulating heavy metals
in their aboveground tissues, without developing any toxicity
symptoms. A metal hyperaccumulator is a plant that can concen-
trate the metals to a level of 0.1% (of the leaf dry weight) for Ni, Co,
Cr, Cu, Al and Pb; 1% for Zn and Mn; and 0.01% for Cd and Se (Baker
and Brooks, 1989; Baker et al., 2000). The time taken by plants to
reduce the amount of heavy metals in contaminated soils depends
on biomass production as well as on their bioconcentration factor
(BCF), which is the ratio of metal concentration in the shoot tissue
to the soil (McGrath and Zhao, 2003). It is determined by the
capacity of the roots to take up metals and their ability to accu-
mulate, store and detoxify metals while maintaining metabolism,
growth and biomass production (Gleba et al., 1999; Guerinot and
Salt, 2001; Clemens et al., 2002). With the exception of hyper-
accumulators, most plants have metal bioconcentration factors of
less than 1, which means that it takes longer than a human lifespan
to reduce soil contamination by 50% (Peuke and Rennenberg,
2005). Hyperaccumulators have a bioconcentration factor greater
than 1, sometimes reaching as high as 50e100. The relationship
between metal hyperaccumulation and tolerance is still a subject
of debate. Views range from no correlation between hyper-
accumulators and the degree of tolerance to metals (Baker et al.,
1994) to strong association between these traits (Chaney et al.,
1997). It is increasingly being realized that to cope with high
concentrations of metals in their tissue, plants must also tolerate
the metals that they accumulate.
There has long been a general agreement that metal hyper-
accumulation is an evolutionary adaptation by specialized plants to
live in habitats that are naturally rich in specic minerals that
confers on them the qualities of increased metal tolerance,
protection against herbivores or pathogens, drought tolerance, and
allelopathy (Boyd and Martens, 1992; Macnair, 1993). The hypoth-
esis of protection against pathogens and herbivores is considered
the most accepted one (Boyd and Martens, 1994; Huitson and
Macnair, 2003; Boyd, 2007; Noret et al., 2007; Galeas et al.,
2008). However, the mechanisms of metal uptake, tolerance to
high metal concentrations, and the exact roles that high level of
elements play in the survival of hyperaccumulators have continued
to be debated.
Hyperaccumulation of heavy metal ions is a striking phenom-
enon exhibited by approximately <0.2% of angiosperms (Baker and
Whiting, 2002; Rascio and Navari-Izzo, 2011). Metal hyper-
accumulators have been reported to occur in over 450 species of
vascular plants from 45 angiosperm families (Table 1) including
members of the Asteraceae, Brassicaceae, Caryophyllaceae, Cyper-
aceae, Cunoniaceae, Fabaceae, Flacourtiaceae, Lamiaceae, Poaceae,
Violaceae, and Euphorbiaceae (Padmavathiamma and Li, 2007), but
are well represented in Brassicaceae especially in the genera
Alyssumand Thlaspi, wherein accumulation of more than one metal
has been reported (Reeves and Baker, 2000; Prasad and Freitas,
2003; Verbruggen et al., 2009; Vamerali et al., 2010) (Table 1).
Pteris vittata (Chinese brake fern) is known to accumulate up to 95%
of the arsenic taken up fromsoil in its fronds (Ma et al., 2001; Zhang
et al., 2002). The best known angiosperm hyperaccumulator of
metals is Thlaspi (now: Noccaea) caerulescens (pennycress), which
can accumulate large amounts of Zn (39,600 mg/kg) and Cd
A. Bhargava et al. / Journal of Environmental Management 105 (2012) 103e120 104
(1800 mg/kg) without apparent damage (Brown et al., 1995; Basic
et al., 2006; Rascio and Navari-Izzo, 2011). This small, self-
pollinating diploid plant can easily grow under lab conditions and
therefore represents an excellent experimental systemfor studying
the mechanisms of metal uptake, accumulation and tolerance in
relation to metal phytoextraction. Apart from T. caerulescens,
Brassica juncea has also been used as a model system to investigate
the physiology and biochemistry of metal accumulation in plants.
2.2. High biomass crops
For successful and economically feasible phytoextraction, it is
necessary to use plants having a metal bioconcentration factor of 20
and a biomass production of 10 tonnes per hectare (t/ha); or plants
with a metal bioconcentration factor of 10 and a biomass produc-
tion of 20 t/ha (Peuke and Rennenberg, 2005). The rate of phy-
toextraction is directly proportional to plant growth rate and the
total amount of metal phytoextracted is correlated with the plant
biomass, which makes the process of phytoextraction very slow
(Shah and Nongkynrih, 2007). This necessitates the identication of
fast growing (largest potential biomass and greatest nutrient
responses) and strongly metal-accumulating genotypes. B. juncea,
while having one-third the concentration of Zn in its tissue, is
considered to be more effective at Zn removal from soil than
T. caerulescens, a known hyperaccumulator of Zn (Ebbs et al., 1997).
This advantage is primarily due to the fact that B. juncea produces
ten-times more biomass than T. caerulescens.
Recently, interest has arisen in the use of high-biomass crops for
phytoextraction of metals (Doty, 2008; Capuana, 2011). Fast-
growing trees are ideal low cost candidates for phytoextraction
due to their extensive root systems, high rates of water uptake and
transpiration, rapid growth, large biomass production and easy
harvesting with subsequent resprouting without disturbance of the
site (Peuke and Rennenberg, 2005). Several tree species have
evoked interest in the phytoremediation of metal contaminated
soils and show great prospects for heavy metal remediation
(Pulford and Watson, 2003; Rosselli et al., 2003; Meers et al., 2007;
Unterbrunner et al., 2007; Brunner et al., 2008; Domnguez et al.,
2008). Poplar and willow, though not hyperaccumulators, are
effective because of their greater biomass and deep root systems,
which makes them effective remediators of metal contamination.
Poplars can be grown in a wide range of climatic conditions and are
used with increasing frequency in short-rotation forestry systems
for pulp and paper production (Peuke and Rennenberg, 2005). This
raises the possibility of using plantations of poplars across several
multiyear cycles to remove heavy metals from contaminated soils.
Importantly, it is unlikely that poplars will enter the human food
chain or end up as feedstock for animals. Likewise, several species
of willow (Salix dasyclados, Salix smithiana and Salix caprea) display
good accumulation capabilities and remediation effectiveness,
Table 1
Important plant species that are metal hyperaccumulators.
Metal Number of hyperaccumulator
species reported
Plant species that accumulate
specic metals
Family Reference
Ni 320 Berkheya coddii Asteraceae Robinson et al. (1997); Moradi et al. (2010)
Alyssum serpyllifolium, A. bertolonii Brassicaceae Becerra-Castro et al. (2009); Barzanti et al. (2011)
Sebertia acuminata Sapotaceae Jaffre et al. (1976); Perrier (2004)
Phidiasia lindavii Acanthaceae Reeves et al. (1999)
Bornmuellera kiyakii Brassicaceae Reeves et al. (2009)
Cu 34 Ipomea alpina Convolvulaceae Cunningham and Ow (1996)
Crassula helmsii Crassulaceae Kpper et al. (2009)
Commelina communis Commelinaceae Wang and Zhong (2011)
Co 34 Haumaniastrum robertii Lamiaceae Brooks (1998)
Crotalaria cobalticola Fabaceae Oven et al. (2002)
Se 20 Astragalus bisulcatus Fabaceae Galeas et al. (2007)
Stanleya pinnata Brassicaceae Freeman et al. (2010); Hladun et al. (2011)
Zn 18 Thlaspi caerulescens Brassicaceae Kupper and Kochian (2010)
Arabis gemmifera, A. paniculata Brassicaceae Kubota and Takenaka (2003); Tang et al. (2009)
Sedum alfredii Crassulaceae Sun et al. (2005)
Arabidopsis halleri Brassicaceae Zhao et al. (2000)
Picris divaricata Asteraceae Du et al. (2011)
Pb 14 Sesbania drummondii Fabaceae Sahi et al. (2002); Sharma et al. (2004)
Hemidesmus indicus Apocynaceae Chandra Sekhar et al. (2005)
Arabis paniculata Brassicaceae Tang et al. (2009)
Plantago orbignyana Plantaginaceae Bech et al. (2011)
Mn 9 Austromyrtus bidwillii Myrtaceae Bidwell et al. (2002)
Phytolacca americana Phytolaccaceae Pollard et al. (2009)
Virotia neurophylla Proteaceae Fernando et al. (2006)
Gossia bidwillii Myrtaceae Fernando et al. (2007)
Maytenus founieri Celastraceae Fernando et al. (2008)
Cd 04 Thlaspi caerulescens Brassicaceae Basic et al. (2006)
Arabidopsis halleri Brassicaceae Dahmani-Muller et al. (2000); Bert et al. (2002)
Bidens pilosa Asteraceae Sun et al. (2009)
Cr na
a
Salsola kali Amaranthaceae Gardea-Torresday et al. (2005)
Leersia hexandra Poaceae Zhang et al. (2007)
Gynura pseudochina Asteraceae Mongkhonsin et al. (2011)
Tl na
a
Iberis intermedia Brassicaceae Leblanc et al. (1999)
Brassica oleracea Brassicaceae Al-Najar et al. (2005)
a
Not available.
similar to herbaceous hyperaccumulators like Arabidopsis halleri
and T. caerulescens, compensating lower metal content in shoots
with higher biomass production (Fischerov et al., 2006; Keller
et al., 2006; Meers et al., 2007). However, the use of perennial
tree species having extensive root systems with elevated metal
content would require excavation and disposal, especially short-
rotation coppice (SRC) after several harvests and at the process
end (Mench et al., 2010).
3. Endophytes and phytoextraction
In the last few years, a lot of research has brought forward the
role of endophytic bacteria in phytoextraction of pollutants (Doty,
2008). Endophytes are the microbes that inhabit the internal of
plant tissues without causing harm to the host (Kuklinsky-Sobral
et al., 2004). Endophytes can facilitate plant growth, increase
resistance of plants to pathogens, drought and herbivores (Bottini
et al., 2004; Saikkonen et al., 2006; Taghavi et al., 2010). Till now
most of the studies have focused on biodegradation of organic
pollutants and applications of endophytic bacteria for improving
phytoremediation of heavy metals. This has been slow because of
lack of valuable strains having the heavy metal resistance and
detoxication capacities (Luo et al., 2011).
Although most of the research on endophytes that assist in
phytoremediation has focused on bacteria, the arbuscular mycor-
rhizal (AM) fungi are also involved in uptake of elements into plants
(Doty, 2008). AM fungi are soil-borne obligate biotrophs that are
integral functioning parts of plant living in mutualistic association
with the roots of about 80% terrestrial plants (Smith and Read,
2008). AM fungi are also reported to be present on the roots of
plants growing on heavy metal-contaminated soils and play an
important role in metal accumulation and tolerance (Gaur and
Adholeya, 2004; Khade and Adholeya, 2009; Javaid, 2011;
Miransari, 2011). An important point about treating polluted soil
with mycorrhizal plants is the selection of appropriate AM species.
The species selected from areas polluted with heavy metals are the
most efcient species which have attained the ability to survive
under metal stress conditions and hence can act more efciently
relative to the other AM species (Miransari, 2011). Mycorrhizal
fungi improve phytoextraction by making metals more available for
uptake by plants. Improved phytoextraction following mycorrh-
ization may be achieved by several mechanisms like increased
tolerance of plants to metals, better plant growth, increased
biomass production and greater metal concentrations in plant
tissues (Vamerali et al., 2010). Generally, species of genus Glomus
are predominant in the rhizosphere of plants growing in heavy
metal-contaminated soils (Khade and Adholeya, 2009; Bedini et al.,
2010). However, the genotypic variation makes it difcult to iden-
tify suitable AM fungi for the restoration of metal-contaminated
soils. Therefore, sustained efforts are required to identify heavy
metal-tolerant mycorrhizal strains for their ultimate application in
the management of metal-contaminated soils (Javaid, 2011).
4. Biochemical mechanisms in accumulators
Plants showing tolerance to toxic metals have a range of
mechanisms at the cellular and molecular level that might be
involved in the general homeostasis, detoxication and tolerance to
heavy metal stress (Hall, 2002). Four processes are generally
believed to be crucial for accumulation: uptake of metals by roots,
transport of metals from roots to shoot, complexation with
chelating molecules and compartmentalization into the vacuole
(Hall, 2002; McGrath and Zhao, 2003). For effective phytoex-
traction, metals must not only be taken up rapidly, but should also
be transported from the roots to aerial parts of the plant. Despite
the presence of large amounts of metals in the soil, the uptake of
metals is mainly inuenced by their bioavailable fraction (Vamerali
et al., 2010). Though abundant in nature, the actual bioavailability
of some metals is limited because of low solubility of metals in
oxygenated water and strong binding to soil particles. Metal
availability and mobility of metals in the rhizosphere is also inu-
enced by rhizospheric microbes as well as the root exudates. With
the exception of Fe, little is known about active mobilization of
trace elements by plant roots. Acidication of the rhizosphere,
exudation of carboxylates and mechanisms assisting in the acqui-
sition of phosphorus contribute to increasing the bioavailability of
certain micronutrients (Chaney et al., 2007). It is assumed that most
of the accumulated metals are bound to ligands like organic acids,
amino acids, peptides and proteins (Verbruggen et al., 2009). While
some aspects of metal detoxication by ligands have been discov-
ered, there is no complete picture of the different chelators
involved in different stages of the internal transport of metals in the
plant and storage in accumulators. There is no conclusive evidence
that hyperaccumulators exude specic chelators in the rhizosphere
to enhance metal uptake. Therefore, the release of specic chelators
associated with enhanced metal uptake and translocation needs
more extensive research (do Nascimento and Xing, 2006).
Several plants are known to possess highly specialized mecha-
nisms to stimulate metal bioavailability in the rhizosphere for
metal uptake. Siderophores are small molecular mass organic
compounds produced by microorganisms and members of family
Poaceae that are capable of enhancing the availability of iron for
uptake into roots (Neubauer et al., 2000; Devez et al., 2009).
Siderophores are specic Fe (III) ligands which form stable
complexes with metals like Cd, Cu, Ni, Pb and Zn (Nair et al., 2007).
The specic chelation of metals can be of great relevance to
decontamination of soil having high metal content. It was sug-
gested that transgenic plants could be developed to secrete metal
selective ligands into the rhizosphere, which could specically
solubilize elements of interest (Raskin, 1996; Neubauer et al., 2000).
The efciency of phytoextraction may be increased by growing
siderophore producing grass species in combination with accu-
mulator plants. Although this approach holds promise, phytosi-
derophores obtain their specicity not by chelation specically only
of Fe in soils, but from their uptake of Fe-phytosiderophores by
a membrane carrier (Parker et al., 1995; Yehuda et al., 1996;
Neubauer et al., 2000). Researchers feel that nding other biosyn-
thetic molecules with selective chelation ability, which plants can
make and secrete into the rhizosphere at adequate concentrations
and create a selective transport protein for the metal chelate seems
difcult, but worth examination to develop unique phytor-
emediation tools. Attempts have been made to identify ligands,
which were believed to be secreted by roots of hyperaccumulators
to increase the rate of Ni release from soil and/or uptake by roots
(Kramer et al., 2000; Pinel et al., 2003; Puschenreiter et al., 2003;
Wenzel et al., 2003a, 2003b). One such study compared the
hyperaccumulator Thlaspi with wheat species that secreted phy-
tosiderophores. The wheat rhizosphere solution contained
substantial levels of ligand(s) as compared to Thlaspi rhizosphere
solution that contained very little (Zhao et al., 2001). Several
researches support the model in which up-regulation or constitu-
tive high activity of element transporters in plasma membranes
allow plants to achieve hyperaccumulation (as opposed to using
secreted ligands) (Chaney et al., 2007). While ligand secretion and
other common root processes do not allow plants to achieve
hyperaccumulation, they can alter elemental speciation in
the rhizosphere and thereby inuence the phytoavailability of
elements in the rhizosphere. Although the concept of making
plants secreting chelating ligands into the rhizosphere and then
absorbing the metaleligand complex into roots remains plausible
for phytoextraction, there is no evidence that such an approach
could be developed into practical phytoextraction systems (Chaney
et al., 2007).
Apart from these natural mechanisms, addition of synthetic
chelates is known to stimulate the release of metals into soil solu-
tion and therefore enhance the potential for uptake into roots (Lasat,
2002). Several chelating agents such as ethylenediaminetetraacetic
acid (EDTA), trans-1, 2-cyclohexylenedinitrilotetraacetic acid
(CDTA), diethylenetriamine-pentaacetic acid (DTPA), ethylene
glycol-O,O
0
-bis-[2-amino-ethyl]-N,N,N
0
,N
0
, tetracetic acid (EGTA),
ethylenediamine di-o-hydroxyphenlyacetic acid (EDDHA), methyl-
glycinediacetate (MGDA), nitrilotriacetic acid (NTA) and citric acid
have been studied for their ability to mobilize metals and increase
metal accumulation in plant species. EDTA is most widely used in
phytoextraction research and has been successfully utilized to
remediate heavy metal contaminated soils (Chen et al., 2004). The
extent of heavy metal solubilization by chelation with organic
complexing agents follows the order of their stability constants,
which are determined in aqueous solutions using the ratio of metal
to chelating agent of 1:1. However despite the success of this
technology, concerns have been raised regarding the enhanced
mobility of metals in soil and their potential risk of leaching to
ground water (Cooper et al., 1999). In addition to the risk of metal
leaching, the use of chelators is also limited by their high cost.
Chelating agents which are more rapidly degraded by soil microbes
yet chelate metals well (eg. EDDS) are more expensive and do not
appreciably decrease metal leaching from treated soil (Meers et al.,
2005). Chaney et al. (2002) estimated that the cost for the amount of
EDTA reportedly needed to attain over 10 g Pb kg
1
dry shoots
would be about $30,000 ha
1
(10 mmol EDTA kg
1
soil for each
cropping).
Following mobilization, metals are rst bound to the cell wall,
which is an ion exchanger of comparatively low selectivity. Trans-
port systems and intracellular high-afnity binding sites then
mediate and drive uptake across the plasma membrane through
secondary transporters such as channel proteins and/or H
-
coupled carrier proteins (Chaney et al., 2007). A number of
important membrane transporter gene families have been identi-
ed and characterized by heterologous complementation screens
and sequencing of ESTs and plant genomes (Table 2). Several cation
transporters have been identied in recent years most of which are
in the ZIP (ZRT, IRT-like protein), nramp (natural resistance-
associated macrophage protein), ysl (Yellow-stripe-like trans-
porter), nas (Nicotinamine synthase), sams (S-adenosyl-methionine
synthetase), fer (Ferritin Fe (III) binding), cdf (cation diffusion
facilitator), hma (heavy metal ATPase) and ireg (Iron regulated
transporter) family (Guerinot, 2000; Williams et al., 2000; Talke
et al., 2006; van de Mortel et al., 2006; Kramer et al., 2007;
Memon and Schroder, 2009; Maestri et al., 2010) (Table 2).
Once inside the plant, further movement of metal containing
sap from roots to the aerial parts is controlled by root pressure as
well as the transpirational pull (Robinson et al., 2003). Metal
transport to the shoot primarily takes place through the xylem.
Efcient translocation of metal ions to the shoot requires radial
symplastic passage and active loading into the xylem (Clemens,
2006; Xing et al., 2008). Due to extreme toxicity of metals at
high intracellular concentrations, plants catalyze redox reactions
and alter the chemistry of these metal ions thereby allowing
their accumulation in non-toxic forms. Typical examples include
reduction of Cr
6
to Cr
3
in Eichornia crassipes (Lytle et al., 1998)
and As
5
to As
3
in B. juncea (Pickering et al., 2000). However, some
of the heavy metals like Pb, Zn and Cd do not occur in different
oxidation states. In some cases, intracellular metal is detoxied via
binding to low molecular mass organic compounds, localization in
the vacuoles as a metal-organic acid complex or by binding to
histidine (Persans et al., 1999; Kramer et al., 2000; Rascio and
Navari-Izzo, 2011). For some metals like Zn, various mechanisms
for regulation of cytoplasmic metal concentration have been put
forward that include sequestration in a subcellular organelle,
complexation to low molecular mass organic ligands, low uptake
Table 2
Important metal transporter genes in different plant species involved in heavy metal tolerance and accumulation.
Family Gene Plant Metal transported Reference
Zn-regulated transporter (ZRT) zip1-12 Arabidopsis thaliana Zn Weber et al. (2004); Roosens et al. (2008a,b)
zip4 Oryza sativa Zn Ishimaru et al. (2005)
zip Medicago truncatula Zn Lopez-Millan et al. (2004)
znt1-2 T. caerulescens Zn van de Mortel et al. (2006)
Fe-regulated transporter (IRT) irt1 Arabidopsis thaliana Fe Kerkeb et al. (2008)
irt1-2 Lycopersicon esculentum Fe Bereczky et al. (2003)
irt1-2 T. caerulescens Fe Schikora et al. (2006); Plaza et al. (2007)
Natural resistance-associated macrophage
proteins (NRAMP)
nramp1-3 Lycopersicon esculentum Fe Bereczky et al. (2003)
nramp4 Thlaspi japonicum Fe Mizuno et al. (2005)
nramp1 Malus baccata Fe Xiao et al. (2008)
Cation diffusion facilitator (CDF) mtp1 Arabidopsis thaliana Zn Kawachi et al. (2008)
mtp1 Arabidopsis halleri Zn Willems et al. (2007)
mtp1 Thlaspi goesingense Zn, Ni Kim et al. (2004)
mtp1 Nicotiana tabacum Zn, Co Shingu et al. (2005)
Al activated malate transporter (ALMT) almt1 Triticum sp. Al Sasaki et al. (2004)
almt1 Secale cereale Al Collins et al. (2008)
P-Type, ATPase (Heavy Metal Associated) hma8 Glycine max Cu Bernal et al. (2007)
hma9 Oryza sativa Cu, Zn, Cd Lee et al. (2007)
hma4 Arabidopsis halleri Cd Courbot et al. (2007)
hma3 Arabidopsis thaliana Co, Zn, Cd, Pb Morel et al. (2008)
Nicotianamine synthase (NAS) nas2, nas3 Arabidopsis halleri Zn Talke et al. (2006)
Copper transporter copt1 Arabidopsis thaliana Cu Sancenon et al. (2004)
Andres-Colas et al. (2010)
Yellow Stripe Like (YSL) ysl2 Arabidopsis thaliana Fe, Cu DiDonato et al. (2004)
ysl3 T. caerulescens Fe, Ni Gendre et al. (2006)
across the plasma membrane, precipitation as insoluble salts and
active extrusion across the plasma membrane into the apoplast
(Brune et al., 1994).
Chelation and sequestration of metals by particular ligands are
important mechanisms used by plants to deal with metal stress.
The two best-characterized metal-binding ligands in plant cells are
metallothioneins (MTs) (Table 3) and phytochelatins (PCs) (Cobbett,
2000; Cobbett and Goldsbrough, 2002). These ligands are widely
distributed in plants, and many physiological and genetic studies
indicate that PCs and MTs are critical for metal tolerance and
accumulation in plants (Hartley-Whitaker et al., 2001; van Hoof
et al., 2001; Cobbett and Goldsbrough, 2002). Both form stable
complexes with metals in the cytosol which can be later seques-
tered into the vacuole (Goldsbrough, 2000). PCs, a family of thiol-
rich peptides, consists of repetitions of the g-Glu-Cys dipeptide
followed by a terminal Gly with the basic structure (g-Glu-Cys)n-
Gly [(PC)n] where n is in the range of two to ve (Memon and
Schroder, 2009). Overproduction of PCs appears to be an induc-
ible rather than a constitutive mechanism, observed especially in
metal non-tolerant plants as part of their defence mechanism
against metals (Freeman et al., 2005). Phytochelatins are also
reported to be involved in arsenic detoxication (Schmoger et al.,
2000). MTs are low molecular mass cysteine rich proteins that
were originally isolated as Cu, Zn and Cd binding proteins in
animals (Gratao et al., 2005; Yang et al., 2009). MTs from plant and
animal sources are being explored for introduction in plants for
reducing metal accumulation in shoots by trapping the metal in
roots (Table 3). The overexpression of MTs can increase plant
tolerance to specic metals. A wide range of MT genes fromvarious
organisms like mouse, hamster, humans, yeast, cyanobacteria and
plants have been overexpressed in plants (Table 3). However, the
evaluation of transgenic plants expressing mammalian genes has
been limited to laboratory assessments only.
The vacuole is generally considered to be the main storage site
for metals in plant cells especially Cd and Zn. Compartmentaliza-
tion of metals in the vacuole is an important part of the tolerance
mechanism of some metal hyperaccumulator plants. The Ni
hyperaccumulator Thlaspi goesingense enhances its Ni tolerance by
compartmentalizing most of the intracellular leaf Ni into the
vacuole (Kramer et al., 2000). High level of metal ion transporter
TgMTP1 in T. goesingense is considered to be the main factor
responsible for the enhanced ability to accumulate metal ions
within shoot vacuoles (Persans et al., 2001). Intact vacuoles isolated
from tobacco and barley exposed to Zn have been shown to accu-
mulate this metal (Burken and Schnoor, 1996).
5. Factors affecting heavy metal uptake by plants
The uptake of heavy metals by plants depends on several factors
some of which are discussed below:
5.1. Soil factors
Several edaphic factors like sorptive capacity of soil, heavy metal
content, cation exchange capacity, soil pH and organic matter
content affect metal hyperaccumulation in plants (Alloway, 1995;
Tiller et al., 1995; Cheng, 2003; Chaney et al., 2007). Generally, only
a fraction of soil metal is readily available (bioavailable) for plant
uptake since the bulk of soil metals are commonly found as insol-
uble compounds unavailable for transport into roots (Lasat, 2002).
Cations of heavy metals are often bound to soil particles because of
soil cation exchange capacity. The cation exchange capacity is
a measure of the soils capacity to exchange ions. The negative
charges are supplied by clay and organic matter of the soil. The
binding afnity of cations reduces cation movement in vascular
plants. Thus, the higher the cation exchange capacity (CEC) of the
soil, the greater the sorption and immobilization of the metals.
Metal solubility and availability are dependent on soil charac-
teristics and are strongly inuenced by soil pH, which is considered
as the major factor inuencing the availability of elements in the
soil for plant uptake. Plants absorb mineral elements in the ionic
form in solution, and the presence of these forms is strongly
inuenced by matrix pH (Dzantor and Beauchamp, 2002). A lower
soil pH increases concentration of heavy metals in the solution by
decreasing their adsorption. In soil, the solution concentrations of
metal contaminants tend to increase with decreasing pHbecause of
their displacement from exchangeable sites on solid surfaces by
increasing activity of hydrogen ions as there is a decrease in pH.
This can increase the availability of the contaminant for plant
uptake, but can also result in concentrations of elements at levels
that are toxic to the plant. Many metal cations like Cd, Cu, Hg, Pb,
and Zn are reported to be more soluble and available in the soil
solution at low pH (below 5.5) (Blaylock and Huang, 2000). The
increased availability of metals at low pH has led phytoextraction
researchers to study incorporation of acidiers (NH
4
containing
fertilizers, organic and inorganic acids, and elemental S) into metal
contaminated soils to improve the success of phytoextraction.
However, inspite of the promise of some acidifying agents, little
research has been carried out on this subject and this needs further
investigation.
The organic content of the soil also has a strong bearing on the
extent of phytoextraction of heavy metals. The addition of peat and
manure is reported to increase Cu, Zn and Ni accumulation inwheat
(Narwal and Singh, 1998). Peat and manure are quite heteroge-
neous substances that can concurrently exert mobilizing and
stabilizing effects (Schmidt, 2003). Acid peat reduces soil pH, which
increases concentration of soluble metals in the soil. It also
increases the CEC of soils, provides sorption sites, reduces metal
mobility and promotes higher binding afnity (Schmidt, 2003).
The oxidation state of a metal contaminant also determines its
solubility and relative availability for uptake by plant systems. In
Table 3
Expression of metallothionein genes in transgenic plants.
MT gene Source Plant species genetically modied Reference
mt-IA Mouse (Mus musculus) Nicotiana tabacum Pan et al. (1994)
mt-b-glucuronidase fusion Chinese hamster (Cricetulus griseus) Nicotiana tabacum Hattori et al. (1994)
mt-II Humans (Homo sapiens) Nicotiana tabacum de Borne et al. (1998)
cup1 Yeast (Saccharomyces cerevisiae) Nicotiana tabacum Thomas et al. (2003)
tymt Cattail (Typha latifolia) Arabidopsis thaliana Zhang et al. (2004)
hmt Humans (Homo sapiens) Medicago varia Watrud et al. (2006)
mt-1 Mouse (Mus musculus) Lycopersicon exculentum Sheng et al. (2007)
smtA Cyanobacteria (Synechococcus sp.) Arabidopsis thaliana Xu et al. (2010)
femt3 Buckwheat (Fagopyrum sp.) Nicotiana debneyii Nikolic et al. (2010)
ccmt1 Pigeon pea (Cajanus cajan) Arabidopsis thaliana Sekhar et al. (2011)
psmtA Pea (Pisum sativum) Populus alba Turchi et al. (2012)
general, the oxidized forms of most common metal contaminants
are less soluble and consequently less available for plant uptake,
chromium being the exception.
5.2. Other environmental factors
The climate at a particular site poses the greatest and most
obvious limitation to the applicability of phytoextraction. Temper-
ature inuences transpiration, water chemistry, growth and
metabolism of plants and therefore, both uptake and elimination of
pollutants (Yu et al., 2005). The removal rates of metals by plants
are known to increase linearly with increase of temperature
(Yu et al., 2010). Baghour et al. (2001) found that potato plants
(Solanum tuberosum L.) showed higher uptake of Cr at high
temperature as compared to plants grown at low temperature. Yu
et al. (2010) investigated the removal of chromium from hydro-
ponic solution by hybrid willows plants to changes in temperature
ranging from 11 to 32

C and found the highest removal rate at
32

C. Temperature dependency of phytoremediation has been one
of the major limitations of the technology, especially in temperate
regions where most eld bioremediation is limited to the warmer
parts of the year. Technologies currently exist for warming up
contaminated matrices to allow bioremediation implementation
during cold periods, but it is questionable whether adoption of such
energy-demanding approaches can still make bioremediation the
favoured, cost effective alternative to other remediation schemes
(Dzantor and Beauchamp, 2002).
Environmental pollution may also exert complex inuence on
mineral accumulation as the plant roots absorb heavy metals from
the soils and aerosols penetrate from the atmosphere into the
plants through the surface of leaves. Minute metal particles stick to
the leaves but only some are absorbed. For example, Pb remains as
surface precipitate, while Cu, Cd and Zn can partially penetrate into
the leaves (Kabata-Pendias and Pendias, 1992).
5.3. Genetic factors
Plant genotype is considered as the most important factor
affecting heavy metal uptake by plants. A range of plant families
distributed over areas diverse geographically, but possessing
a common characteristic of natural enrichment for some specic
metal is known (Prasad and Freitas, 2003). At the species level,
numerous reports have conrmed the existence of signicant
genotypic differences in the heavy metal uptake and distribution
between and within species, and even within cultivars (Chardot
et al., 2005; Liu et al., 2005; Grant et al., 2007; Bhargava et al.,
2008a; Richau and Schat, 2009; Kramer, 2010; Hanikenne and
Nouet, 2011). Some genotypes respond positively to increased
heavy metal concentration in soil, while others may be inert or
show negative growth. In some cases, both the genomes of an
amphidiploids acting together are responsible for high metal
accumulation, like the AABB genome in B. juncea (Nanda-Kumar
et al., 1995).
The existence of metal accumulators demonstrates that some
plants have the genetic potential to clean contaminated soils. It has
been demonstrated that tolerance to high levels of metals by
hyperaccumulators is under genetic control (Macnair, 1993), which
allows the plants to produce specic molecules that react with
metals to form complexes. These complexes can then be stored
away from sensitive tissues. Understanding the genetic mechanism
of metal accumulation in hyperaccumulator species is important
because it facilitates the use of various approaches to genetic
improvement of plants for metal uptake. The genetic control of
heavy metal accumulation in plants is not well understood. Earlier
works suggested that metal tolerance is complex and is the result of
the action of a large number of genes (Antonovics et al., 1971).
However, recent studies have shown that metal tolerance is regu-
lated by a few major genes or sometimes by a single gene (Macnair,
1993; Clarke, 1995; Macnair et al., 2000). The hybridization of Zn
tolerant and non-tolerant species has brought out the fact that both
traits are genetically independent, and Zn tolerance is controlled by
a single gene. Several researchers have conducted interesting
genetic studies with the species A. halleri that is considered close to
Arabidopsis thaliana and has undergone a natural selection for Zn
tolerance (Bert et al., 2000; Roosens et al., 2008a, 2008b). A. halleri
is a Zn hyperaccumulator that is small in size and hyper-
accumulates Zn from Zn rich soils (Table 2). Zn tolerance and
hyperaccumulation have been found to be constitutive in A. halleri
(Chaney et al., 2007). Macnair et al. (1999) made crosses between
A. halleri and Arabidopsis lyrata subsp. petraea and subsequent
backcrosses to allow evaluation of the inheritance. From the anal-
ysis of the F
2
generation it was found that Zn hyperaccumulation
and tolerance were separate genetic properties under independent
genetic control in A. halleri. Bert et al. (2003) analyzed the co-
segregation of Cd tolerance and Cd accumulation in the progeny
from an A. halleri A. lyrata subsp. petraea F
1
plant backcrossed to
A. lyrata subsp. petraea (BC1). It was found that these characters
inherited independently, but the presence of co-segregation of Cd
tolerance with Zn tolerance, and Cd accumulation with Zn accu-
mulation suggested partial pleiotropic control of these characters.
Thus, Zn tolerance appeared to be controlled by a single gene, while
Cd and Zn hyperaccumulation was controlled by several genes
(Macnair et al., 1999; Bert et al., 2003). Evaluation of differences
between A. halleri accessions has shown small variation in metal
accumulation, but no apparent difference in inheritance of metal
accumulation (Bert et al., 2000).
Isolation of the quantitative trait loci (QTL) associated with
metal tolerance holds great promise for the identication of the
genes responsible for this adaptation (Roosens et al., 2008a, 2008b).
Mapping of quantitative trait loci (QTLs) for Zn and Cd tolerance has
shown 3 additive QTLs for Zn and Cd tolerance, with trait-
enhancing alleles originating from the A. halleri parent (Courbot
et al., 2007; Willems et al., 2007). The co-location of 1 QTL for Cd
tolerance with a Zn tolerance locus has conrmed the hypothesis of
partial pleiotropic control of Zn and Cd tolerance (Bert et al., 2003).
Deniau et al. (2006) mapped QTLs for Cd and Zn accumulation in an
F
2
cross between 2 plants and found 1 common locus for Cd and Zn
in root, 1 for Cd in root and Cd in shoot, and 1 for Zn in root and Zn
in shoot. The trait-enhancing alleles at the 3 Cd accumulation loci
were all derived from the parent having greater Cd accumulation
capacity, while both parents contributed trait-enhancing alleles to
the Zn accumulation loci. Deniau et al. (2006) found transgressive
segregation for Zn accumulation, but not for Cd accumulation. The
accumulation rates of Zn and Cd in the F
2
plants were signicantly
phenotypically correlated and governed by common genetic
determinants, as well as additionally by more metal-specic
determinants. Segregation studies in F
3
and F
4
families derived
froma cross betweenT. caerulescens accessions have shown that Zn
and Ni accumulation are pleiotropically controlled by the same
genes (Richau and Schat, 2009). The high-throughput technologies
like microarray support the idea that genes that are thought to be
involved in hyperaccumulation and hypertolerance are not species-
specic or novel, but rather differently expressed and regulated,
compared with non-hyperaccumulator species (Verbruggen et al.,
2009).
6. Plant improvement for enhanced phytoextraction
The goal of remediating metal contaminated soil is generally to
extract the metal from the large soil volume and transfer it to
a smaller volume of plant tissue for harvest and disposal. This is
due to the fact that metals cannot be metabolized or broken down
to less toxic forms. The amount of pollutant a plant can remove
from the soil is a function of its tissue concentration multiplied by
the quantity of biomass formed (Macek et al., 2007). Lowyield and
slow growth rates have been cited as limiting factors for the
development of effective metal phytoremediator plants (Brown
et al., 1995). Most of the known metal accumulating plants are
metal selective, show slow growth rate, produce relatively little
biomass and can be used for phytoextraction in their natural
habitats only (Kamnev and van der Lelie, 2000). Thus, while the
amounts of metal concentration per unit of plant biomass can be
high, the total amounts of metal removed at a site during a given
period can be quite low. For example, although T. caerulescens can
take up sufcient levels of metals to make harvesting and metal
recovery economical, they are often limited by their small biomass
(Meagher et al., 2000; Nedelkoska and Doran, 2000). Moreover,
the use of hyperaccumulator plants can be limited because of
less information about their agronomic characteristics, pest
management, breeding potential and physiological processes
(Cunningham et al., 1995). However, a rapidly growing non-
accumulator could be modied to enable it to achieve some
of the properties of hyperaccumulators. Two approaches are
currently being explored to develop and/or improve the metal
accumulating plants:
1) Conventional breeding, and 2) genetic engineering.
6.1. Conventional breeding
Conventional breeding approaches coupled with suitable
agronomic practices like soil fertilization and conditioning, proper
plant density, crop rotation, weed control and irrigation practices
can go a long way in enhancing the phytoextraction capacity vis-
-vis metals (Lasat, 2000). Traditional plant breeding uses the
available genetic diversity within a species to combine the traits
needed for successful phytoextraction (Li et al., 2003). Although
there have been efforts to study genotypic differences in metal
uptake and biomass in hyperaccumulator species, little effort has
been made to breed these species for domestication. Efcient
management and utilization of germplasm requires detailed
knowledge of the genetic diversity of agronomic traits for proper
characterization of populations to facilitate efcient synthesis of
breeding populations that are designed to accomplish specic
objectives (Bhargava et al., 2007, 2008a). In any crop-breeding
program, assessment of genetic diversity is invaluable as it helps
in the identication of diverse parental combinations to create
segregating progenies with maximum genetic variability and
facilitates introgression of desirable genes from diverse germ-
plasm into the available genetic base (Bhargava et al., 2007). The
existing genetic diversity in crops can be used for phytoextraction
by identifying easily cultivable, high biomass yielding plants and
practicing selection in future generations. Numerous reports are
available which suggest the manipulation of germplasm and use
of selection strategies to reduce heavy metal concentration in
different plant species (Li et al., 1997; Liu et al., 2005; Bhargava
et al., 2008a). Apart from selection strategy, conventional
breeding methodology also involves the transfer of hyper-
accumulator phenotype like metal tolerance and increased uptake
from small, slow growing, hyperaccumulator species to fast
growing, high biomass producing non-accumulator plants
through hybridization (Chaney et al., 2000; Grant et al., 2007;
Bhargava et al., 2008a, 2008b).
Researchers all over the world are looking for newplant species
having natural variability for heavy metal accumulation. Recently,
there have been reports on the utilization of the available genetic
diversity in underutilized crops for phytoextraction (Lasat et al.,
1998; Tamura et al., 2005; Bhargava et al., 2008a). Common
buckwheat (Fagopyrum esculentum Moench) is the rst known Pb
hyperaccumulator with high biomass productivity (Tamura et al.,
2005). F. esculentum is reported to naturally accumulate up to
4200 mg/g of Pb in the shoot. Amending the soil with methyl-
glycinediacetate (MGDA), a biodegradable chelator, resulted in
a 5-fold increase in the Pb shoot concentration. These ndings
qualify this species as an excellent candidate for remediating
Pb-contaminated soils. The red root pigweed (Amaranthus retro-
exus L.) is known to accumulate soil
137
Cs to levels which may
support a phytoextraction technology, while other crop plants are
less able to accumulate Cs (Lasat et al., 1998). Studies conducted on
another underutilized pseudocereal, Chenopodium, indicated
a large genetic variation for various heavy metals at species and
sub-species levels, coupled with high biomass (Bhargava et al.,
2008a). At the species level, Chenopodium quinoa showed high
accumulation of Zn, Cr and Cd, while Chenopodium album and the
American species Chenopodium bushianum accumulated high
amounts of Ni (Bhargava et al., 2008a). This genetic diversity could
be effectively exploited in breeding crops with high phytoex-
traction properties.
Somaclonal variation offers the possibility to obtain changes in
one or a few characters of an otherwise outstanding cultivar
without altering the remaining and often unique part of the
genotype. One future line of action can be to utilize somaclonal
variation for development of plants having improved phytoex-
traction capacity. Symmetric and asymmetric somatic hybridiza-
tions have recently gained importance as genetic modiers.
Brewer et al. (1999) raised somatic hybrids (protoplast electro-
fusion) between Zn hyperaccumulator T. caerulescens and Brassica
napus and found that the hybrids accumulated high levels of Zn
that would otherwise have been toxic to B. napus. Some of the
hybrids produced high biomass combined with high metal accu-
mulation and tolerance, making them attractive for Zn phytoex-
traction. This study clearly indicates that transfer of metal
hyperaccumulating phenotype is feasible. Likewise, Gleba et al.
(1999) also obtained somatic hybrids from a cross between
B. juncea and T. caerulescens that had signicant Pb accumulating
properties. Metal resistant traits were introduced into the high-
biomass Pb accumulator B. juncea using somatic hybridization.
T. caerulescens, a known Ni and Zn hyperaccumulator, was selected
as one of the parents for both symmetric and asymmetric hybrids
in which T. caerulescens protoplasts were irradiated before fusion.
Eighteen hybrids were regenerated of which 2 were found to be
fertile. One of these hybrids had vigorous growth, characteristic of
B. juncea and increased resistance when grown in Pb, Ni, and Zn
contaminated soil. The total amount of Pb phytoextracted by each
hybrid plant was much greater because of the large biomass
produced on the contaminated soil. Several other researches have
also reported the utilization of somatic hybridization to introduce
toxic metal resistant traits present in T. caerulescens into B. juncea
(Dushenkov et al., 2002; Alkorta et al., 2004). The hybrids so
obtained have demonstrated high metal accumulation potential,
tolerance to toxic metals, and good biomass production. In vitro
breeding and somaclonal variation have been used to improve the
potential of Indian mustard (B. juncea) to extract and accumulate
toxic metals (Nehnevajova et al., 2007). Calli from B. juncea were
cultivated on a modied MS medium and somaclones were
regenerated from metal-tolerant callus cells. Seven out of thirty
individual variants showed higher metal extraction than the
control plants. The improvement of metal shoot accumulation of
the best regenerant and metal extraction indicated that B. juncea
could be used for phytoremediation purpose (Nehnevajova et al.,
2007).
6.2. Genetic engineering
Several anatomical constraints severely restrict sexual compat-
ibility between taxa and pose serious limitations in developing
hybrids with increased phytoextraction capability. Biotechnology
has opened new gateways in phytoremediation technology by
offering the opportunity for direct gene transfer and overcoming
the limitations imposed by sexual incompatibility. The develop-
ment of transgenic plants with increased metal uptake, accumu-
lation and tolerance to toxicity is nowbeing considered a promising
alternative (Fig. 1). Genetic engineering is a technique that can be
applied advantageously to the development for ideal phytor-
emediation plants that combine high metal accumulating capacity
and high aboveground biomass yield (Krenlampi et al., 2000).
With the use of genetic engineering, it is feasible to manipulate
a plants capacity to tolerate, accumulate, and/or metabolize
pollutants, and to create an ideal plant for environmental cleanup.
Recent progress in determining the molecular basis of metal
accumulation and tolerance by hyperaccumulators has provided us
with a strong scientic basis to outline some strategies for
achieving this goal. Many genes are reported to be involved in
metal uptake, translocation, sequestration, chemical modication,
and tolerance (Table 4). The introduction and overexpression of the
hyperaccumulating genes into a non-hyperaccumulator plant could
be a possible way to enhance metal uptake, accumulation, tolerance
and detoxication process (Clemens et al., 2002). The over-
expression of genes encoding the rate-limiting gene product is
expected to lead a faster overall rate of the pathway and to more
efcient phytoremediation (Pilon-Smits and Pilon, 2002). Besides
this, the repression of an endogenous gene by inserting a gene of
reverse orientation (antisense technology) can also result in
enhanced metal uptake by plants (Shah and Nongkynrih, 2007).
The introduction of an additional metal binding domain to the
implemented protein further enhances the metal binding capacity
(Macek et al., 1996; Kotrba et al., 1999).
The fact that plants can benet from organic acid exudation in
a number of ways has aroused interest of biotechnologists to
increase organic acid exudation in crop and pasture species (Ryan
et al., 2001). Several reports on transgenic plants tolerant to the
presence of toxic levels of metals have appeared in recent
years (Table 5). In most of the studies, the overexpression of the
genes encoding for the enzymes of phytochelatin synthase, ACC
deaminase, S-metabolism, glutathione, Hg
2
-reductase, arsenate
reductase, aldolase/aldehyde reductase, enzymes of histidine
biosynthesis and metallothionein (MT)-genes have been effectively
carried out (Shah and Nongkynrih, 2007). The bioengineering of
transporter genes to manipulate the transport of metal ions inside
the cell has also been successfully exploited and a combination of
these genes in rapidly growing plant species has led to promising
results.
Several strategies have been successfully used to create trans-
genics that show promising properties for phytoremediation.
Table 5 provides an overview of the different approaches involved
in the production of transgenics for enhancing phytoextraction
capacity of crop plants for different heavy metals. One strategy for
increasing the efciency of phytoextraction involves increase in the
metal translocation to the shoot by increasing plant transpiration.
Gleba et al. (1998) observed that the genetically modied plants of
B. juncea with an increased transpiration phytoextract 104% more
lead than the wild type plants, making it a good candidate for eld
optimization and use. A simple and direct method for enhancing
the effectiveness of phytoextraction is to overexpress in transgenic
plants, the genes involved in metabolism, uptake, or transport of
specic pollutants. The introduction of these genes has been
successfully achieved using Agrobacterium tumefaciens-mediated
Search for metal accumulators/organisms
living in high metal stress
Isolation and cloning of genes conferring
metal tolerance and/or accumulation
Introduction of gene in the target plant;
Field trials
Study organism at
low and high metal
stress
Comparison with
close relatives
Characterization of
gene products in a
suitable model system
Introduction of the gene
in a model plant; Stress
induced studies
Transgenic plant for phytoremediation
Fig. 1. Development of metal tolerant/accumulator plant using genetic engineering (Adapted from Krenlampi et al., 2000).
Table 4
Some important genes involved in metal tolerance and accumulation.
Metal Gene Reference
Al aha2 Moffat (1999)
Fe frd3, Durrett et al. (2007)
Zn mtp1, mtp3 Gustin et al. (2009)
Cu cbf Szira et al. (2008)
Cd atcax2, atcax 4, mt1 Pan et al. (1994); Korenkov et al. (2007)
Ni reg2, sat-c Schaaf et al. (2006); Na and Salt (2011)
plant transformation. The overexpression of glutamylcysteine
synthetase has been accomplished by genetic engineering in Pop-
ulus angustifolia, Nicotiana tabacum and Silene cucubalus which
enhances heavy metal accumulation as compared to the wild type
plants (Fulekar et al., 2009). Tomato plants have been genetically
modied to express 1-amino-cyclopropane-1-carboxylic acid (ACC)
deaminase, a bacterial gene which resulted in enhanced metal
tolerance and increased uptake of a range of heavy metals (Grichko
et al., 2000). The overexpression of Escherichia coli gshI gene
encoding g-glutamylcysteine synthetase in B. juncea resulted in
increased concentration of Cd (up to 90%) in transgenic plants as
compared to wild types (Zhu et al., 1999). Another promising
approach is to overexpress enzymes catalyzing rate-limiting steps.
ATP sulfurylase (APS) is a rate-limiting enzyme in the selenium
detoxication process which facilitates the reduction of selenate to
selenite. The overexpression of an ATP sulfurylase (APS) from
A. thaliana in B. juncea led to three times more uptake and accu-
mulation of metals in the transgenic plants as compared to the wild
types (Pilon-Smits et al., 1999). In another study, Indian mustard
plants overexpressing cystathionine gamma synthase (CGS) were
developed. It was observed that the transgenics had enhanced
tolerance to selenite and volatilized Se two to three times faster
thanwild type, while at the same time accumulating less Se in roots
and shoots (van Huysen et al., 2003). However, inspite of the initial
success with B. juncea, the plant has never been shown to be useful
in eld phytoextraction. It has poor yield and rapidly owers if
grown under warm conditions. Also the plant has no metal toler-
ance and no metal accumulation in absence of added chelators.
Modication or overexpression of the enzymes that are involved
in the synthesis of PCs is a promising approach to enhance heavy
metal tolerance and accumulation in plants and has been tried to
genetically transform high biomass plants into efcient phytor-
emediators (Zhu et al., 1999). Several attempts have been made to
increase the formation of PCs by overexpressing genes encoding
enzymes that could stimulate the synthesis of cysteine and gluta-
thione. The induction of PCs by Cd suggests that increasing
biosynthesis of phytochelatins would improve tolerance and phy-
toextraction, but there has been no supporting evidence that this
approach would yield a Cd phytoextraction plant. Cd tolerance has
been increased to a maximum 3e7 fold by high expression of PC
synthase (Heiss et al., 2003), which is trivial as compared to the 200
times higher tolerance of Zn and Cd of T. caerulescens (Chaney et al.,
2005; Wang et al., 2006). Similar ndings have been observed
when researchers tried to alter Cd accumulation in tobacco shoots
by transgenic expression of phytochelatin or metallothioneins,
despite extensive testing of many constructs (Lugon-Moulin et al.,
2004).
For some metals such as Hg and selenium, a promising strategy
is to convert the metal to a volatile form for release and dilution
into the atmosphere. The most spectacular application of biotech-
nology in phytoremediation has been the engineering of plants
capable of removing methyl-Hg from contaminated soils (Rugh
et al., 1996; Pilon-Smits and Pilon, 2000). Arabidopsis plants were
the rst to be engineered with a gene, mercuric ion reductase
(merA), from a bacterium that was resistant to Hg (Rugh et al.,
1996). Later, transgenic Arabidopsis plants were raised wherein
merB enzyme was targeted to the endoplasmic reticulum
(Bizily et al., 2003), which showed a 10e70 times higher specic
activity to degrade organic Hg than transgenic plants with
cytoplasmic merB. This was followed by transformation of yellow
poplar (Liriodendron tulipifera) and Eastern cottonwood (Populus
deltoides) with merA, leading to increased tolerance to ionic Hg
(Rugh et al., 1998; Che et al., 2003). Transgenic cottonwood
shoots had normal growth on medium containing 25 mM Hg (II),
a concentration of Hg that killed the wild-type shoots. The trans-
genic plants produced up to 4-fold more elemental Hg than wild-
type plants, demonstrating that the plants take up and transform
Hg to the less toxic form. Successful transformation of rice (Oryza
sativa) has been achieved for enhanced Hg remediation (Heaton
et al., 2003). The merA transgenic rice tolerated concentrations of
Hg
2
that killed the wild-type controls and steadily converted the
Hg
2
to its less toxic volatile form (Heaton et al., 2003).
Table 5
Transgenic approaches for enhancement of heavy metal uptake in different plants.
Metal Plant Methodology/Approach Reference
Ni Brassica napus Arabidopsis Expression of bacterial ACC deaminase gene Stearns et al. (2005)
Expression of nicotianamine synthase cDNA (TcNAS1) Pianelli et al. (2005)
Cd Brassica juncea Overexpression of gamma-glutamylcysteine synthetase and glutathione synthetase Bennett et al. (2003)
Reisinger et al. (2008)
Arabidopsis Overexpression of Arabidopsis phytochelatin synthase (AtPCS1) Lee et al. (2003)
Arabidopsis Expression of yeast ABC transporter family member, YCF1 Song et al. (2003)
Pb Arabidopsis Expression of yeast ABC transporter family member, YCF1 Song et al. (2003)
Brassica juncea Overexpression of a yeast cadmium factor 1 (YCF1) Bhuiyan et al. (2011a)
Brassica juncea Overexpression of ATP-binding cassette (ABC) transporter gene AtATM3 Bhuiyan et al. (2011b)
Cu Nicotiana tabacum Expression of yeast metallothionein (CUP1) gene Thomas et al. (2003)
Populus alba Expression of P
S
MT
A1
gene from Pisum sativum Balestrazzi et al. (2009)
Arabidopsis Expression of yeast copper-dependent transcription factor ACE1 Xu et al. (2009)
Zn Arabidopsis Overexpression of Zn Induced Facilitator 1 (ZIF 1) Haydon and Cobbett (2007)
Brassica juncea Overproduction of the g-glutamylcysteine synthetase and glutathione synthetase Bennett et al. (2003)
Nicotiana tabacum Overexpression of glyoxalase pathway enzymes Singla-Pareek et al. (2006)
Hg Arabidopsis Expression of bacterial gamma-glutamylcysteine synthetase gene under control
of a strong constitutive actin regulatory sequence (A2)
Li et al. (2005)
Nicotiana tabacum Expression of ppk gene specied bacterial polyphosphate (polyP) Nagata et al. (2006)
Nicotiana tabacum Expression of a modied bacterial Hg reductase, merA gene Heaton et al. (2005)
As Arabidopsis Expression of bacterial gamma-glutamylcysteine synthetase gene under control
of a strong constitutive actin regulatory sequence (A2)
Li et al. (2005)
Brassica juncea Overexpression of ATP sulfurylase Wangeline et al. (2004)
Brassica napus Expression of Enterobacter cloacae UW4 1-aminocyclopropane-1-carboxylate (ACC)
deaminase (EC 4.1.99.4) gene
Nie et al. (2002)
Bizily et al. (2000) accomplished the full pathway from methyl
Hg to the least toxic metallic Hg by expressing both merA and
organomercurial lyase (merB) within the same plant. Arabidopsis
plants transformed with both genes were tolerant to concentra-
tions of methyl-Hg 50 times higher than the concentrations to
which wild-type plants were tolerant and 10 times higher than
those to which plants transformed with merB alone were tolerant.
Transformation of Spartina alterniora, the common wetland
plant, with both merA and merB has been achieved, resulting in
increased tolerance of the transgenic plants to phenylmercuric
acetate and mercuric chloride (Czako et al., 2006). Recently,
Eastern cottonwood was transformed with both merA and merB
(Lyyra et al., 2007). These transgenic cottonwoods were strongly
resistant to toxic phenylmercuric acetate, and had an increased
rate of detoxication that was 2e3 times faster than that of the
control plants. In the transgenics, merB catalyzes the protonlysis of
the CeHg bond with the release of Hg
2
, which is subsequently
converted by merA to Hg
0
, a less toxic volatile element that is
released into the atmosphere. Such genetically modied plants
release 10 times of elemental Hg as compared to non-transformed
plants.
Following the transgenic approach, only Hg has been demon-
strated to work in the eld (Heaton et al., 2003), but public
acceptance has been difcult because Hg
0
is volatilized at the soil
surface and will eventually be re-deposited on soil or water
(Chaney et al., 2007). Unfortunately, the contaminant was not
destroyed but merely transformed from soil-bound ionic Hg
2
to
airborne elemental Hg
0
.
Clonal propagation has opened the door to the creation of tree
remediation cultivars i.e. perennial trees having excellent metal
uptake (Stomp et al., 1993). Work is underway to screen tree species
for their ability to tolerate, take up, translocate, sequester, and
detoxify heavy metal ions. Clone stability and in vitro phytoex-
traction capacity of vegetative clones of Populus canescens that
included two transgenic clones (ggs11 and lgl6) were studied as
in vitro leaf disc cultures (Gyulai et al., 2005). The transgenic poplar
cyt-ECS (ggs11) clone, as stimulated by the presence of Zn, showed
elevated heavy metal (Cu) uptake as compared to the non-
transformed clone. This suggests that gshI-transgenic poplars
may be suitable for phytoremediation of soils contaminated with
Zn and Cu (Gyulai et al., 2005). Future research may focus on
optimizing metal accumulation in cloned plants and developing
clones of trees that exhibit a high degree of accumulation and larger
biomass.
7. Limitations
Phytoextraction seems to hold promise for the remediation of
metal polluted soils. Nevertheless, ongoing research reveals that
the applicability of the technique is debatable since the practical
implications are not so evident (van Nevel et al., 2007). Presently,
the technology is limited due to long period required for cleanup,
restricted number of target metals that can be extracted, limited
depth that can be assessed by the roots, decline in phytoextraction
efciency under increasing metal concentrations and the lack of
knowledge on the agronomic practices and management (Keller
et al., 2003; Ernst, 2005; McGrath et al., 2006; Robinson et al.,
2006; Audet and Charest, 2007). Also, the complexity of hyper-
accumulation has not been fully understood, either at the tissue or
at the subcellular level. Phytoextraction seems possible for only As
and Ni, while for the other metals the technology still appears to be
far from practice. The vast majority of the hyperaccumulator
species discovered so far are Ni hyperaccumulators (Table 1), while
species accumulating Cu, Pb, Cd, Zn, Co and As are much less
numerous (do Nascimento and Xing, 2006).
Most of the data on the performance of phytoremediating
transgenic plants are based on observations made in controlled
conditions (laboratory) often on growth media, rather than in the
eld. Therefore, it is important to conrm the performance of
phytoremediation systems on large-scale contaminated sites. The
bioavailability of the contaminants on the contaminated sites
appears to be a major factor in the discrepancy between lab and
eld conditions. A better understanding of soil properties and the
physio-chemical factors inuencing the solubility of toxic
compounds is likely to help in the improvement of on-site plant
performances in the future (Singh et al., 2003). There is also an
urgent need to gain indepth knowledge about the molecular
mechanisms that allow plants to remediate polluted soils, partic-
ularly with respect to hyperaccumulation and hypertolerance. The
identication of genes involved in the acquisition and the homeo-
stasis of toxic materials, along with an understanding of the way
they are regulated is likely to improve phytoremediation tech-
nology to a great extent (Cluis, 2004).
8. Is phytoextraction commercially viable?
Metal-contaminated soil can be remediated by physical, chem-
ical or biological techniques. Chemical and physical treatments of
metal contaminated soils irreversibly affect soil properties, destroy
biodiversity and may render the soil useless as a medium for plant
growth (Padmavathiamma and Li, 2007). There is a need to develop
suitable cost-effective biological soil remediation techniques to
remove contaminants without affecting soil fertility. Phytoex-
traction is one of the low cost techniques for contaminated soil
remediation (Table 6).
In the absence of long-termsite trials, phytoextraction efciency
should be evaluated using models. Phytoextraction projects can be
evaluated on the amount of metals being removed from the soil in
relation to the total amount present in the soil (Mertens et al., 2005,
2006) and on the time needed for remediation. However, assess-
ment of feasibility of phytoextraction based on modelling has its
own limitations. It has been suggested that the biomass production
might decrease over time due to nutrient depletion in the soil after
several croppings or pest infections (van Nevel et al., 2007). To
mitigate this, fertilizers, pest control and crop rotation might be
necessary. In natural conditions, plant growth may be limited by
other environmental variables, such as low pH, low water avail-
ability, salinity or insufcient aeration, whereas experimental
conditions are generally optimal. Lastly, a large area of metal-
contaminated soils is polluted with more than one element,
while few plant species can extract high concentrations of more
than one element (Ernst, 2005). This polymetallicity strongly
affects the productivity even of metal resistant plants, causing the
extraction period to become too long to be economically feasible
(Ernst, 2005; Robinson et al., 2006).
Development of a commercially viable technology for using
hyperaccumulator plant species to phytoextract metals requires the
identication or creation of an ideal phytoextraction plant, opti-
mization of soil and crop management practices, and development
Table 6
Cost comparison of different cleanup techniques for metal
pollution (Glass, 1999b).
Process Cost (US $/t)
Land lling 100e500
Vitrication 75e425
Chemical treatment 100e500
Electrokinetics 20e200
Phytoextraction 5e40
of methods for biomass processing and extraction (Li et al., 2003).
Phytoextraction can be applied either to metal-contaminated soils
or to ores that cannot be economically extracted by traditional
mining technology. In the former case, phytoextraction is a type of
phytoremediation, while the term phytomining has been applied
to the latter case where the economic value of the recovered metal
is the primary motive (Li et al., 2003). The value of the phytoex-
tracted metal, if it is recycled, can help to defray the costs of phy-
toremediation. Phytomining includes the generation of revenue by
extracting soluble metals produced by the plant biomass ash, also
known as bio-ore. If phytoextraction could be combined with
biomass generation and is commercially utilized as an energy
source, then it could be turned into a protable operation, with the
residual ash available to be used as an ore (Cunningham and Ow,
1996; Brooks, 1998; Padmavathiamma and Li, 2007). With some
metals like Ni, Zn and Cu, the value of reclaimed metal may provide
an additional incentive for phytoremediation (Chaney et al., 1997;
Thangavel and Subhuram, 2004). The total world remediation
market is reported to be approximately US $15e18 billion per year
(Memon and Schroder, 2009). Several companies and research
groups around the world are pursuing phytomining strategies.
B. juncea and other plant species are reported to accumulate as
much as 20 mg/kg of gold in greenhouse experiments when plants
were supplemented with a solubilizing agent (ammonium thiocy-
anate) (Prasad, 2003). Some commercial companies are gaining
economic benets from phytoextraction not just by recovering
extracted metals fromplant biomass, but also by using the biomass
for energy generation (Glass, 1999a, 2000). There is evidence for
extraction of high purity Ni fromNi-contaminated Alyssumbiomass
(Bani et al., 2007; Chaney et al., 2008). The commercially viable
options include either the recycling of Ni rich biomass in metal-
lurgy or use as Ni fertiliser to correct Ni deciency in eld crops. A
signicant number of companies based on phytoextraction have
emerged that have created growing industries in North America
and in several European countries. Smaller, but emerging markets
exist in developing nations, particularly in portions of Asia. There
are about 10 companies in Canada and 20 in Europe which are
conducting research or which have carried out commercial reme-
diation using phytoremediation or related technologies. Numerous
commercial projects have been undertaken in several countries
around the world though some of these companies are pursuing
phytoremediation at the research stage only (Glass, 2000).
9. Prospects
The advantages of using metal accumulating plants for the
removal of metals from contaminated soils include lower costs,
generation of recyclable metal-rich plant residue, applicability to
a range of toxic metals, minimal environmental disturbance and
public acceptance. Plants with increased metal accumulation
properties may also be utilized to enhance crop productivity in
areas with suboptimal metal levels, or as fortied food and feed
(Guerinot and Salt, 2001). But, inspite of these advantages, phy-
toextraction is still an emerging technology and concerted efforts
are needed if this environmental friendly technology is to be
exploited.
The sequencing of complete genome of hyperaccumulators
could go a long way in identifying promising functional noncoding
regions and to narrow the focus for experimental tests (Wray and
Babbitt, 2008). The discovery of metal related genes with the aid
of genome sequencing will open up newavenues for the creation of
transgenics having desired properties that would help in estab-
lishing phytoextraction as a potent technology for environmental
cleanup. The understanding of genome evolution in the hyper-
accumulators should be improved by merging ecological and
molecular genomics (Verbruggen et al., 2009). The search for
signatures of recent adaptive evolution across candidate genes for
metal tolerance or accumulation could be a promising approach.
Apart from constitutive overexpression of a single gene, several
genes may be expressed simultaneously in specic cellular
components under specic conditions. Several approaches could be
followed to achieve this goal. These include identication of the
metal transporter proteins and introducing genes encoding trans-
porter molecules to enhance the ability of metal ions to enter plant
cells (Tong et al., 2004); overexpression of the enzyme phytoche-
latin synthase (PS) in plants that would lead to increased metal
tolerance and accumulation; overproduction of nicotianamine to
manipulate metal translocation and tolerance as well as iron uptake
in cereals (Pilon-Smits and Pilon, 2002); overproduction of histi-
dine (His) since the genes involved in His biosynthesis have been
cloned (Persans et al., 1999) and preliminary studies suggesting
that histidine overproducing plants have enhanced Ni tolerance
(Kramer and Chardonnens, 2001).
However, the use of transgenics in nature for phytoextraction
should be preceded by a thorough risk assessment study on a case-
by-case basis and weighing the benets and risks as compared to
alternative technologies. For phytoremediation to be a viable, cost
effective alternative, plants will need to alter or detoxify the
contaminants rather than simply accumulate or displace them.
Theoretical assessment of risks associated with the use of metal-
volatilizating plants has pointed that the use of transgenic plants
having phytoextraction capacity is relatively safe (Lin et al., 2000;
Meagher et al., 2000; Rugh et al., 2000). Still, concerns have been
raised about the safety of phytoextraction that range from metals
entering the food chain through herbivores, accumulation of metals
in the topsoil and dispersion of plant material to adjacent envi-
ronments (Perronnet et al., 2000; Linacre et al., 2003; Mertens
et al., 2005, 2007). Although the risk of transgenic plants or their
genes escaping into the environment is not considered to be
a signicant problem, the transgenic gene frequency should be
analyzed for a number of generations over polluted and non-
contaminated soils by a greenhouse or pilot eld experiment
(Pilon-Smits and Pilon, 2002). To further minimize the risk of
outcrossing to wild relatives, the use of transgenics should be
undertaken in consonance with classical breeding approaches.
Transgenic plant species should be chosen that have no compatible
wild relatives, male-sterile transgenics may be bred, and the plants
may be harvested before owering (Pilon-Smits and Pilon, 2002).
Field trials with transgenic poplars were carried out in former
copper-mining regions of Russia (Middle Urals, Swerdlovsk oblast)
and Germany (Saxonia Anhalt, district Mansfelder Land) to assess
the biosafety risk of transgenic poplars developed for the remedi-
ation of contaminated soils by elucidating the stability of the
transgene under actual eld conditions and the possibility of
horizontal gene transfer to microorganisms present in the rhizo-
sphere. Preliminary results showed that the transgenic poplars
were genetically stable with no indications of any impact on the
environment (Peuke and Rennenberg, 2005).
10. Conclusion
Phytoremediation is an environment friendly green technology
involving living plants which offers a cost-effective means for
cleaning metal-contaminated soils. In order to exploit its full
potential, a comprehensive understanding is needed on as to how
metal uptake, transport, and trafcking across plant membranes
and distribution, tolerance, sensitivity, etc., take place under
different environments (Aruna Kumara, 2011). The technology is
more applicable to soils or ores that cannot be economically
enriched by traditional mining. Biotechnology would enable the
identication of new genes to tackle the issue of environmental
cleanup. Several crop species are being researched upon and await
incorporation of such genes not only for cleaning up the heavy
metals but also to extend the area under cultivation. The challenge
facing phytoextraction should be faced jointly by scientists, envi-
ronmental engineers and science administrators to prove the
technologys efcacy at pilot sites. It is incumbent upon researchers
to conduct basic laboratory work as well as follow newapproaches
for identifying and solving diverse scientic issues posed by metal
phytoextraction.
References
Alkorta, I., Hernandez-Allica, J., Becerril, J.M., Amezaga, I., Albizu, I., Garbisu, I., 2004.
Recent ndings on the phytoremediation of soils contaminated with environ-
mentally toxic heavy metals and metalloids such as Zn, Cd, Pb and arsenic.
Environ. Sci. Biotechnol. 3, 71e90.
Alloway, B.J., 1995. Heavy Metals in Soils. Blackie, London.
Al-Najar, H., Kaschl, A., Schulz, R., Romheld, V., 2005. Effects of thallium fractions in
the soil and pollution origin in thallium uptake by hyperaccumulator plants:
a key factor for assessment of phytoextraction. Intern. J. Phytoremed. 7, 55e67.
Andres-Colas, N., Perea-Garcia, A., Puig, S., Penarrubia, L., 2010. Deregulated copper
transport affects Arabidopsis development especially in the absence of
environmental cycles. Plant Physiol. 153, 170e184.
Antonovics, J., Bradsham, A.D., Turner, R.G., 1971. Heavy metal tolerance in plants.
Adv. Ecol. Res. 7, 1e85.
Aruna Kumara, K.K.I.U., 2011. Use of crop plants for removal of heavy metals. In:
Khan, M.S., Zaidi, A., Goel, R., Musarrat, J. (Eds.), Biomanagement of Metal-
Contaminated Soils. Springer, New York, pp. 439e457.
Audet, P., Charest, C., 2007. Heavy metal phytoremediation from a meta-analytical
perspective. Environ. Poll. 147, 231e237.
Babula, P., Adam, V., Opatrilova, R., Zehnalek, J., Havel, L., Kizek, R., 2008.
Uncommon heavy metals, metalloids and their plant toxicity: a review. Environ.
Chem. Lett. 6, 189e213.
Baghour, M., Moreno, D.A., Hernndez, J., Castilla, N., Romero, L., 2001. Inuence of
root temperature on phytoaccumulation of As, Ag, Cr and Sb in potato plants
(Solanum tuberosum L. var. Spunta). J. Environ. Sci. Health (A) 36, 1389e1401.
Baker, A.J.M., Brooks, R.R., 1989. Terrestrial higher plants which hyperaccumulate
metallic elements. A review of their distribution, ecology and phytochemistry.
Biorecovery 1, 81e126.
Baker, A.J.M., Whiting, S.N., 2002. In search of the Holy Grail e a further step in
understanding metal hyperaccumulation? New Phytol. 155, 1e7.
Baker, A.J.M., Reeves, R.D., Hajar, A.S.M., 1994. Heavy metal accumulation and
tolerance in British populations of the metallophyte Thlaspi caerulescens J. and
C. Presl (Brassicaceae). New Phytol. 127, 61e68.
Baker, A.J.M., McGrath, S.P., Reeves, R.D., Smith, J.A.C., 2000. Metal hyper-
accumulator plants: a review of the ecology and physiology of a biological
resource for phytoremediation of metal polluted soils. In: Terry, N., Banuelos, G.
(Eds.), Phytoremediation of Contaminated Soil and Water. Lewis Publishers,
Boca Raton, pp. 85e107.
Baker, A.J.M., 1981. Accumulators and excluders e strategies in the response of
plants to heavy metals. J. Plant Nutr. 3, 643e654.
Balestrazzi, A., Botti, S., Zelasco, S., Biondi, S., Franchin, C., Calligari, P., Racchi, M.,
Turchi, A., Lingua, G., Berta, G., Carbonera, D., 2009. Expression of the PsMT
A1
gene in white poplar engineered with the MAT system is associated with heavy
metal tolerance and protection against 8-hydroxy-20 deoxyguanosine
mediated-DNA damage. Plant Cell Rep. 28, 1179e1192.
Bani, A., Echevarria, G., Sulce, S., Morel, J.L., Mullai, A., 2007. In-situ phytoextraction
of Ni by a native population of Alyssum murale on an ultramac site (Albania).
Plant Soil 293, 79e89.
Barzanti, R., Colzi, I., Arnetoli, M., Gallo, A., Pignattelli, S., Gabbrielli, R., Gonnelli, C.,
2011. Cadmium phytoextraction potential of different Alyssum species.
J. Hazard. Mater. 196, 66e72.
Basic, N., Keller, C., Fontanillas, P., Vittoz, P., Besnard, G., Galland, N., 2006. Cadmium
hyperaccumulation and reproductive traits in natural Thlaspi caerulescens
populations. Plant Biol. 8, 64e72.
Becerra-Castro, C., Monterroso, C., Garcia-Leston, M., Prieto-Fernandez, A.,
Acea, M.J., Kidd, P.S., 2009. Rhizosphere microbial densities and trace
metal tolerance of the nickel hyperaccumulator Alyssum serpyllifolium subsp.
lusitanicum. Int. J. Phytoremed. 11, 525e541.
Bech, J., Duran, P., Roca, N., Poma, W., Snchez, I., Barcel, J., Boluda, R., Roca-
Prez, L., Poschenrieder, C., 2011. Shoot accumulation of several trace elements
in native plant species from contaminated soils in the Peruvian Andes.
J. Geochem. Explor.. doi:10.1016/j.gexplo.2011.04.007
Bedini, S., Turrini, A., Rigo, C., Argese, E., Giovannetti, M., 2010. Molecular charac-
terization and glomalin production of arbuscular mycorrhizal fungi colonizing
a heavy metal polluted ash disposal island, downtown Venice. Soil Biol.
Biochem. 42, 758e765.
Bennett, L.E., Burkhead, J.L., Hale, K.L., Terry, N., Pilon, M., Pilon-Smits, E.A., 2003.
Analysis of transgenic Indian mustard plants for phytoremediation of metal-
contaminated mine tailings. J. Environ. Qual. 32, 432e440.
Bereczky, Z., Wang, H.Y., Schubert, V., Ganal, M., Bauer, P., 2003. Differential regu-
lation of Nramp and IRT metal transporter genes in wild type and iron uptake
mutants of tomato. J. Biol. Chem. 278, 24697e24704.
Bernal, M., Testillano, P.S., Alfonso, M., Del Carmen, R.M., Picorel, R., Yruela, I., 2007.
Identication and subcellular localization of the soybean copper P1B-ATPase
GmHMA8 transporter. J. Struct. Biol. 158, 146e158.
Bert, V., Macnair, M.R., de Laguerie, P., Saumitou-Laprade, P., Petit, D., 2000. Zn
tolerance and accumulation in metallicolous and nonmetallicolous populations
of Arabidopsis halleri (Brassicaceae). New Phytol. 146, 225e233.
Bert, V., Bonnin, I., Saumitou-Laprade, P., de Laguerie, P., Petit, D., 2002. Do
Arabidopsis halleri from nonmetallicolous populations accumulate zinc and
cadmium more effectively than those from metallicolous populations? New
Phytol. 155, 47e57.
Bert, V., Meerts, P., Saumitou-Laprade, P., Salis, P., Gruber, W., Verbruggen, N., 2003.
Genetic basis of Cd tolerance and hyperaccumulation in Arabidopsis halleri.
Plant Soil 249, 9e18.
Bhargava, A., Shukla, S., Rajan, S., Ohri, D., 2007. Genetic diversity for morphological
and quality traits in quinoa (Chenopodium quinoa Willd.) germplasm. Genet.
Res. Crop Evol. 54, 167e173.
Bhargava, A., Shukla, S., Srivastava, J., Singh, N., Ohri, D., 2008a. Genetic diversity for
mineral accumulation in the foliage of Chenopodium spp. Scientia Hortic. 118,
338e346.
Bhargava, A., Shukla, S., Srivastava, J., Singh, N., Ohri, D., 2008b. Chenopodium:
a prospective plant for phytoextraction. Acta Physiol. Plant 30, 111e120.
Bhargava, A., Gupta, V.K., Singh, A.K., Gaur, R., 2012. Microbes for heavy metal
remediation. In: Gaur, R., Mehrotra, S., Pandey, R.R. (Eds.), Microbial Applica-
tions. I.K. International Publ., New Delhi, pp. 167e177.
Bhuiyan, M.S.U., Min, S.R., Jeong, W.J., Sultana, S., Choi, K.S., Song, W.Y., Lee, Y.,
Lim, Y.P., Liu, J.R., 2011a. Overexpression of a yeast cadmium factor 1 (YCF1)
enhances heavy metal tolerance and accumulation in Brassica juncea. Plant Cell
Tiss. Org. Cult. 105, 85e91.
Bhuiyan, M.S.U., Min, S.R., Jeong, W.J., Sultana, S., Choi, K.S., Lee, Y., Liu, J.R., 2011b.
Overexpression of AtATM3 in Brassica juncea confers enhanced heavy metal
tolerance and accumulation. Plant Cell Tiss. Org. Cult. 107, 69e77.
Bidwell, S.D., Woodrow, I.E., Batianoff, G.N., Sommer-Knudsen, J., 2002. Hyper-
accumulation of manganese in the rainforest tree Austromyrtus bidwillii
(Myrtaceae) from Queensland, Australia. Funct. Plant Biol. 29, 899e905.
Bizily, S.P., Rugh, C.L., Meagher, R.B., 2000. Phytodetoxication of hazardous
organomercurials by genetically engineered plants. Nat. Biotech. 18, 213e217.
Bizily, S.P., Kim, T., Kandasamy, M.K., Meagher, R.B., 2003. Subcellular targeting of
methylHg lyase enhances its specic activity for organic Hg detoxication in
plants. Plant Physiol. 131, 463e471.
Blaylock, M.J., Huang, J.W., 2000. Phytoextraction of metals. In: Raskin, I.,
Ensley, B.D. (Eds.), Phytoremediation of Toxic Metals: Using Plants to Clean-up
the Environment. John Wiley, New York, pp. 53e70.
Bottini, R., Cassan, F., Piccoli, P., 2004. Gibberellin production by bacteria and its
involvement in plant growth promotion and yield increase. Appl. Microbiol.
Biotechnol. 65, 497e503.
Boyd, R.S., Martens, S.N., 1992. The raison dtre for metal hyperaccumulation by
plants. In: Baker, A.J.M., Proctor, J., Reeves, R.D. (Eds.), The Vegetation of
Ultramac (Serpentine) Soils. Intercept, Andover, UK, pp. 279e289.
Boyd, R.S., Martens, S.N., 1994. Ni hyperaccumulated by Thlaspi montanum var.
montanum is acutely toxic to an insect herbivore. Oikos 70, 21e25.
Boyd, R.S., 2007. The defense hypothesis of elemental hyperaccumulation: status,
challenges and new directions. Plant Soil 293, 153e176.
Brewer, E.P., Saunders, J.A., Angle, J.S., Chaney, R.L., McIntosh, M.S., 1999. Somatic
hybridization between the Zn accumulator Thlaspi caerulescens and Brassica
napus. Theor. Appl. Genet. 99, 761e771.
Brooks, R.R., 1998. Plants That Hyperaccumulate Heavy Metals. CAB Intern.,
Wallingford.
Brown, S.L., Chaney, R.F., Angle, J.S., Baker, A.J.M., 1995. Zn and Cd uptake by
hyperaccumulator Thlaspi caerulescens and metal tolerant Silene vulgaris, grown
on sludge-amended soils. Environ. Sci. Tech. 29, 1581e1585.
Brune, A., Urbach, W., Dietz, K.J., 1994. Compartmentation and transport of Zn in
barley primary leaves as basic mechanisms involved in Zn tolerance. Plant Cell
Environ. 17, 153e162.
Brunner, I., Luster, J., Gnthardt-Goerg, M.S., Frey, B., 2008. Heavy metal accumu-
lation and phytostabilisation potential of tree ne roots in a contaminated soil.
Environ. Poll. 152, 559e568.
Burken, J.G., Schnoor, J.L., 1996. Phytoremediation: plant uptake of atrazine and role
of root exudates. J. Environ. Eng. 122, 958e963.
Capuana, M., 2011. Heavy metals and woody plants-biotechnologies for phytor-
emediation. iForest 4, 7e15.
Chandra Sekhar, K., Kamala, C.T., Chary, N.S., Balaram, V., Garcia, G., 2005. Potential
of Hemidesmus indicus for phytoextraction of lead from industrially contami-
nated soils. Chemosphere 58, 507e514.
Chaney, R.L., Malik, M., Li, Y.-M., Brown, S.L., Brewer, E.P., Angle, J.S., Baker, A.J.M.,
1997. Phytoremediation of soil metals. Curr. Opin. Biotech. 8, 279e284.
Chaney, R.L., Li, Y.-M., Angle, J.S., Baker, A.J.M., Reeves, R.D., Brown, S.L., Homer, F.A.,
Malik, M., Chin, M., 2000. Improving metal hyperaccumulator wild plants to
develop commercial phytoextraction systems: approaches and progress. In:
Terry, N., Bauelos, G.S. (Eds.), Phytoremediation of Contaminated Soil and
Water. CRC Press, Boca Raton, FL, pp. 131e160.
Chaney, R.L., Brown, S.L., Li, Y.-M., Angle, J.S., Stuczynski, T.I., Daniels, W.L.,
Henry, C.L., Siebielec, G., Malik, M., Ryan, J.A., Compton, H., 2002. Progress in
risk assessment for soil metals, and in-situ remediation and phytoextraction of
metals from hazardous contaminated soils. In: Proc. USEPA Conf.
Phytoremediation: State of the Science. 1e2 May 2000, Boston, MA. USEPA,
Washington, DC.
Chaney, R.L., Angle, J.S., McIntosh, M.S., Reeves, R.D., Li, Y.-M., Brewer, E.P.,
Chen, K.-Y., Roseberg, R.J., Perner, H., Synkowski, E.C., Broadhurst, C.L.,
Wang, S., Baker, A.J.M., 2005. Using hyperaccumulator plants to phytoextract
soil Ni and Cd. Z. Naturforsch. 60C, 190e198.
Chaney, R.L., Angle, J.S., Broadhurst, C.L., Peters, C.A., Tappero, R.V., Sparks, D.L.,
2007. Improved understanding of hyperaccumulation yields commercial
phytoextraction and phytomining technologies. J. Environ. Qual. 36,
1429e1443.
Chaney, R.F., Angle, J.S., Baker, A.J.M., Reeves, R.D., Roseberg, R.J., Simmons, R.W.,
Broadhurst, C.L., 2008. Phytoextraction and phytomining of Ni and Cd from
contaminated or mineralized soils. In: Bert, V. (Ed.), COST Action 859, Phyto-
technologies in Practice: Biomass Production, Agricultural Methods, Legacy,
Legal and Economic Aspects. Verneuil-en-Halatte, France, pp. 15e16.
Chardot, V., Massoura, S.T., Echevarria, G., Reeves, R.D., Morel, J.L., 2005. Phytoex-
traction potential of the Ni hyperaccumulators Leptoplax emarginata and
Bornmuellera tymphaea. Int. J. Phytoremed. 7, 323e335.
Che, D., Meagher, R.B., Heaton, A.C., Lima, A., Rugh, C.L., Merkle, S.A., 2003.
Expression of mercuric ion reductase in Eastern cottonwood (Populus deltoides)
confers mercuric ion reduction and resistance. Plant Biotech. J. 1, 311e319.
Chen, Y., Li, X.D., Shen, Z.G., 2004. Leaching and uptake of heavy metals by ten
different species of plants during an EDTA-assisted phytoextraction process.
Chemosphere 57, 187e196.
Cheng, S., 2003. Heavy metals in plants and phytoremediation. Environ. Sci. Poll.
Res. 10, 335e340.
Clarke, J.M., 1995. Inheritance of Grain Cd Concentration in Four Durum Wheat
Crosses. Agronomy Abstracts. American Society of Agronomy, Madison,
Wisconsin.
Clemens, S., Palmgren, M.G., Krmer, U., 2002. A long way ahead: understanding
and engineering plant metal accumulation. Trends Plant Sci. 7, 309e315.
Clemens, S., 2006. Toxic metal accumulation, responses to exposure and mecha-
nisms of tolerance in plants. Biochimie 88, 1707e1719.
Cluis, C., 2004. Junk-greedy greens: phytoremediation as a new option for soil
decontamination. Bioteach J. 2, 61e67.
Cobbett, C., Goldsbrough, P., 2002. Phytochelatins and metallothioneins: roles in
heavy metal detoxication and homeostasis. Ann. Rev. Plant Biol. 53, 159e182.
Cobbett, C.S., 2000. Phytochelatins and their roles in heavy metal detoxication.
Plant Physiol. 123, 825e832.
Collins, N.C., Shirley, N.J., Saeed, M., Pallotta, M., Gustafson, J.P., 2008. An ALMT1
gene cluster controlling aluminum tolerance at the Alt4 locus of rye (Secale
cereale L.). Genetics 179, 669e682.
Cooper, E.M., Sims, J.T., Cunningham, S.D., Huang, J.W., Berti, W.R., 1999. Chelate-
assisted phytoextraction of Pb from contaminated soils. J. Environ. Qual. 28,
1709e1719.
Courbot, M., Willems, G., Motte, P., Arvidsson, S., Roosens, N., Saumitou-Laprade, P.,
Verbruggen, N., 2007. A major quantitative trait locus for Cd tolerance in
Arabidopsis halleri colocalizes with HMA4, a gene encoding a heavy metal
ATPase. Plant Physiol. 144, 1052e1065.
Cunningham, S.D., Ow, D.W., 1996. Promises and prospects of phytoremediation.
Plant Physiol. 110, 715e719.
Cunningham, S.D., Berti, W.R., Huang, J.W., 1995. Phytoremediation of contaminated
soils. Tibtech 13, 393e397.
Czako, M., Feng, X., He, Y., Liang, D., Marton, L., 2006. Transgenic Spartina alterniora
for phytoremediation. Environ. Geochem. Health 28, 103e110.
Dahmani-Muller, H., van Oort, F., Gelie, B., Balabane, M., 2000. Strategies of heavy
metal uptake by three plant species growing near a metal smelter. Environ. Poll.
109, 231e238.
de Borne, F.D., Elmayan, T., de Roton, C., de Hys, L., Tepfer, M., 1998. Cadmium
partitioning in transgenic tobacco plants expressing a mammalian metal-
lothionein gene. Mol. Breed. 4, 83e90.
de Vos, C.H.R., Schat, H., de Waal, M.A.M., Voojs, R., Ernst, W.H.O., 1991. Increased
resistance to Cu-induced damage of root cell plasmalemma in Cu tolerant Silene
cucubalus. Physiol. Plant. 82, 523e528.
Deniau, A., Pieper, B., Ten Bookum, W.M., Lindhout, P., Aarts, M.G.M., Schat, H., 2006.
QTL analysis of Cd and Zn accumulation in the heavy metal hyperaccumulator
Thlaspi caerulescens. Theoret. Appl. Genet. 113, 907e920.
Devez, A., Achterberg, E., Gledhill, M., 2009. Metal ion-binding properties of phy-
tochelatins and related ligands. Metal Ions Life Sci. 5, 441e481.
DiDonato, R.J., Roberts, L.A., Sanderson, T., Eisley, R.B., Walker, E.L., 2004.
Arabidopsis Yellow Stripe-Like2 (YSL2): a metal-regulated gene encoding
a plasma membrane transporter of nicotianamineemetal complexes. Plant J. 39,
403e414.
do Nascimento, C.W.A., Xing, B., 2006. Phytoextraction: a review on enhanced metal
availability and plant accumulation. Sci. Agric. 63, 299e311.
Domnguez, M.T., Maran, T., Murillo, J.M., Schulin, R., Robinson, B.H., 2008. Trace
element accumulation in woody plants of the Guadiamar Valley, SW Spain:
a large-scale phytomanagement case study. Environ. Poll. 152, 50e59.
Doty, S.L., 2008. Enhancing phytoremediation through the use of transgenics and
endophytes. New Phytol. 179, 318e333.
Du, R.J., He, E.K., Tang, Y.T., Hu, P.J., Ying, R.R., Morel, J.L., Qiu, R.L., 2011. How
phytohormone IAA and chelator EDTA affect lead uptake by Zn/Cd hyper-
accumulator Picris divaricata. Int. J. Phytoremed 13, 1024e1036.
Durrett, T.P., Gassmann, W., Rogers, E.E., 2007. The FRD3-mediated efux of citrate
into the root vasculature is necessary for efcient iron translocation. Plant
Physiol. 144, 197e205.
Dushenkov, S., Skarzhinskaya, M., Glimelius, K., Gleba, D., Raskin, I., 2002. Bioen-
gineering of a phytoremediation plant by means of somatic hybridization. Int. J.
Phytoremed. 4, 117e126.
Dzantor, E.K., Beauchamp, R.G., 2002. Phytoremediation, Part I: fundamental basis
for the use of plants in remediation of organic and metal contamination.
Environ. Prac. 4, 77e87.
Ebbs, S.D., Lasat, M.M., Brandy, D.J., Cornish, J., Gordon, R., Kochian, L.V., 1997. Heavy
metals in the environment: phytoextraction of cadmium and zinc from
a contaminated soil. J. Environ. Qual. 26, 1424e1430.
Ernst, W.H.O., 2005. Phytoextraction of mine wastes e options and impossibilities.
Chem. Erde 65 (S1), 29e42.
Fernando, D.R., Bakkaus, E.J., Perrier, N., Baker, A.J.M., Woodrow, I.E., Batianoff, G.N.,
Collins, R.N., 2006. Manganese accumulation in the leaf mesophyll of four tree
species: a PIXE/EDAX localization study. New Phytol. 171, 751e758.
Fernando, D.R., Woodrow, I.E., Bakkaus, E.J., Collins, R.N., Baker, A.J.M.,
Batainoff, G.N., 2007. Variability of Mn hyperaccumulation in the Australian
rainforest tree Gossia bidwillii (Myrtaceae). Plant Soil 293, 145e152.
Fernando, D.R., Woodrow, I.E., Jaffre, T., Dumontet, V., Marshall, A.T., Baker, A.J.M.,
2008. Foliar manganese accumulation by Maytenus founieri (Celastraceae) in its
native New Caledonian habitats: populational variation and localization by
X-ray microanalysis. New Phytol. 177, 178e185.
Fischerov, Z., Tlustos, P., Szkov, J.,

Sichorova, K., 2006. A comparison of phytor-
emediation capability of selected plant species for given trace elements.
Environ. Poll 144, 93e100.
Freeman, J.L., Garcia, D., Kim, D., Hopf, A., Salt, D.E., 2005. Constitutively elevated
salicylic acid signals glutathione-mediated Ni tolerance in Thlaspi Ni hyper-
accumulators. Plant Physiol. 137, 1082e1091.
Freeman, J.L., Tamaoki, M., Stushnoff, C., 2010. Molecular mechanisms of selenium
tolerance and hyperaccumulation in Stanleya pinnata. Plant Physiol. 153,
1630e1652.
Freitas, H., Prasad, M.N.V., Pratas, J., 2004. Plant community tolerant to trace
elements growing on the degraded soils of Sao Domingos mine in the south
east of Portugal: environmental implications. Environ. Int. 30, 65e72.
Fulekar, M.H., Singh, A., Bhaduri, A.M., 2009. Genetic engineering strategies for
enhancing phytoremediation of heavy metals. African J. Biotech. 8, 529e535.
Galeas, M.L., Zhang, L.H., Freeman, J.L., Wegner, M., Pilon-Smits, E.A.H., 2007.
Seasonal uctuations of selenium and sulfur accumulation in selenium hyper-
accumulators and related nonaccumulators. New Phytol. 173, 517e525.
Galeas, M.L., Klamper, E.M., Bennett, L.E., Freeman, J.L., Kondratieff, B.C., Quinn, C.F.,
Pilon-Smits, E.A., 2008. Selenium hyperaccumulation reduces plant arthropod
loads in the eld. New Phytol. 177, 715e724.
Gardea-Torresday, J.L., De la Rosa, G., Peralta-Videa, J.R., Montes, M.,
Cruz-Jimenez, G., Cano-Aguilera, I., 2005. Differential uptake and transport of
trivalent and hexavalent chromium by tumble wed (Salsola kali). Arch. Environ.
Contam. Toxicol. 48, 225e232.
Gaur, A., Adholeya, A., 2004. Prospects of arbuscular mycorrhizal fungi in phytor-
emediation of heavy metal contaminated soils. Curr. Sci. 86, 528e534.
Gendre, D., Czernic, P., Conjro, G., Pianelli, K., Briat, J.-F., Lebrun, M., Mari, S., 2006.
TcYSL3, a member of the YSL gene family from the hyperaccumulator Thlaspi
caerulescens, encodes a nicotianamine-Ni/Fe transporter. Plant J. 49, 1e15.
Glass, D.J., 1999a. US and International Markets for Phytoremediation, 1999e2000.
D.J. Glass Associates, Needham.
Glass, D.J., 1999b. Economic potential of phytoremediation. In: Raskin, I., Ensley, B.D.
(Eds.), Phytoremediation of Toxic Metals: Using Plants to Clean up the
Environment. John Wiley & Sons Inc, New York, pp. 15e31,.
Glass, D.J., 2000. The 2000 Phytoremediation Industry. D.J. Glass Associates,
Needham.
Gleba, D., Gleba, Y., Skarzhinskaya, M., Glimelius, K., Raskin, I., 1998. Transfer of
resistance to heavy metals from Thlaspi to Brassica by asymmetric hybridiza-
tion, Abstract of IXth International Cong. on Plant Tissue Culture, Jerusalem. 145
pp.
Gleba, D., Borisjuk, N.V., Borisjuk, L.G., Kneer, R., Poulev, A., Skarzhinskaya, M.,
Dushenkov, S., Logendra, S., Gleba, Y.Y., Raskin, I., 1999. Use of plant roots for
phytoremediation and molecular farming. Proc. Natl. Acad. Sci. (USA) 96,
5973e5977.
Goldsbrough, P., 2000. Metal tolerance in plants: the role of phytochelatins. In:
Terry, N., Banuelos, G. (Eds.), Phytoremediation of Contaminated Soil and Water.
Lewis Publishers, Boca Raton, pp. 221e233.
Grant, C.A., Clarke, J.M., Duguid, S., Chaney, R.L., 2007. Selection and breeding of
plant cultivars to minimize Cd accumulation. Sci. Total Environ 390, 301e310.
Gratao, P.L., Prasad, M.N.V., Cardoso, P.F., Lea, P.J., Azevedo, R.A., 2005.
Phytoremediation: green technology for the clean up of toxic metals in the
environment. Braz. J. Plant Physiol. 17, 53e64.
Grichko, V.P., Filby, B., Glick, B.R., 2000. Increased ability of transgenic plants
expressing the bacterial enzyme ACC deaminase to accumulate Cd, Co, Cu, Ni,
Pb and Zn. J. Biotech. 81, 45e53.
Guerinot, M.L., Salt, D.E., 2001. Fortied foods and phytoremediation: two sides of
the same coin. Plant Physiol. 125, 164e167.
Guerinot, M.L., 2000. The ZIP family of metal transporters. Biochim. Biophys. Acta
1465, 190e198.
Gustin, J.L., Loureiro, M.E., Kim, D., Na, G., Tikhonova, M., Salt, D.E., 2009.
MTP1-dependent Zn sequestration into shoot vacuoles suggests dual roles in Zn
tolerance and accumulation in Zn-hyperaccumulating plants. Plant J. 57,
1116e1127.
Gyulai, G., Humphreys, M., Bittsanszky, A., Skot, K., Kiss, J., Skotc, L., Gullner, G.,
Heywood, S., Szabo, Z., Lovatt, A., Radimszky, L., Roderick, H., Rennenberg, H.,
Abberton, M., Komives, T., Heszky, L., 2005. AFLP analysis and improved
phytoextraction capacity of transgenic gshI-poplar clones (Populus canescens
L.) for Cu in vitro. Z. Naturforsch. 60, 300e306.
Hall, J.L., 2002. Cellular mechanisms for heavy metal detoxication and tolerance.
J. Exp. Bot. 53, 1e11.
Hanikenne, M., Nouet, C., 2011. Metal hyperaccumulation and hypertolerance:
a model for plant evolutionary genomics. Curr. Opin. Plant Biol. 14, 252e259.
Hartley-Whitaker, J., Ainsworth, G., Vooijs, R., Ten Brookum, W., Schat, H.,
Meharg, A.A., 2001. Phytochelatins are involved in differential arsenate
tolerance in Holcus lanatus. Plant Physiol. 126, 299e306.
Hattori, J., Labbe, H., Miki, B.L., 1994. Construction and expression of a metal-
lothionein-b-glucuronidase gene fusion. Genome 37, 508e512.
Haydon, M.J., Cobbett, C.S., 2007. A novel major facilitator superfamily protein at the
tonoplast inuences Zn tolerance and accumulation in Arabidopsis. Plant
Physiol. 143, 1705e1719.
Heaton, A.C.P., Rugh, C.L., Kim, T., Wang, N.J., Meagher, R.B., 2003. Toward detoxi-
fying Hg-polluted aquatic sediments with rice genetically engineered for Hg
resistance. Environ. Toxicol. Chem. 22, 2940e2947.
Heaton, A.C.P., Rugh, C.L., Wang, N.J., Meagher, R.B., 2005. Physiological responses
of transgenic merA-tobacco (Nicotiana tabacum) to foliar and root mercury
exposure. Water Air Soil Poll. 161, 137e155.
Heiss, S., Wachter, A., Bogs, J., Cobbett, C., Rausch, T., 2003. Phytochelatin synthase
(PCS) protein is induced in Brassica juncea leaves after prolonged Cd exposure.
J. Exp. Bot. 54, 1833e1839.
Hladun, K.R., Parker, D.R., Trumble, J.T., 2011. Selenium accumulation in the oral
tissues of two Brassicaceae species and its impact on oral traits and plant
performance. Environ. Exp. Bot. 74, 90e97.
Huitson, S.B., Macnair, M.R., 2003. Does Zn protect the Zn hyperaccumulator
Arabidopsis halleri from herbivory by snails? New Phytol. 159, 453e459.
Ishimaru, Y., Suzuki, M., Kobayashi, T., Takahashi, M., Nakanishi, H., Mori, S.,
Nishizawa, N.K., 2005. OsZIP4, a novel zinc-regulated zinc transporter in rice.
J. Exp. Bot. 56, 3207e3214.
Jabeen, R., Ahmad, A., Iqbal, M., 2009. Phytoremediation of heavy metals: physio-
logical and molecular mechanisms. Bot. Rev. 75, 339e364.
Jaffre, T., Brooks, R.R., Lee, J., Reeves, R.D., 1976. Sebertia acuminata: a hyper-
accumulator of nickel from New Caledonia. Science 193, 579e580.
Javaid, A., 2011. Importance of arbuscular mycorrhizal fungi in phytoremediation of
heavy metal contaminated soils. In: Khan, M.S., Zaidi, A., Goel, R., Musarrat, J.
(Eds.), Biomanagement of Metal-Contaminated Soils. Springer, New York,
pp. 125e141.
Ji, G.L., Wang, J.H., Zhang, X.N., 2000. Environmental problems in soil and
groundwater induced by acid rain and management strategies in China. In:
Huang, P.M., Iskandar, I.K. (Eds.), Soils and Groundwater Pollution and Reme-
diation. CRC Press, London, pp. 201e224.
Kabata-Pendias, A., Pendias, H., 1992. Trace Elements in Soil and Plants. CRC Press,
Boca Raton, FL.
Kamnev, A.A., van der Lelie, D., 2000. Chemical and biological parameters as tools to
evaluate and improve heavy metal phytoremediation. Biosci. Rep. 20, 239e258.
Krenlampi, S., Schat, H., Vangronsveld, J., Verkleij, J.A.C., van der Lelie, D.,
Mergeay, M., Tervahauta, A.I., 2000. Genetic engineering in the improvement of
plants for phytoremediation of metal polluted soils. Environ. Poll. 7, 225e231.
Kawachi, M., Kobae, Y., Mimura, T., Maeshima, M., 2008. Deletion of a histidine-rich
loop of AtMTP1, a vacuolar Zn
2
/H
antiporter of Arabidopsis thaliana, stimu-

lates the transport activity. J. Biol. Chem. 283, 8374e8383.
Keller, C., Hammer, D., Kayser, A., Richner, W., Brodbeck, M., Sennhauser, M., 2003.
Root development and heavy metal phytoextraction efciency: comparison of
different plant species in the eld. Plant Soil 249, 67e81.
Keller, C., Diallo, S., Cosio, C., Basic, N., Galland, N., 2006. Cadmium tolerance and
hyperaccumulation by Thlaspi caerulescens populations grown in hydroponics
are related to plant uptake characteristics in the eld. Func. Plant Biol. 33,
673e684.
Kerkeb, L., Mukherjee, I., Chatterjee, I., Lahner, B., Salt, D.E., Connolly, E.L., 2008.
Iron-induced turnover of the Arabidopsis Iron-Regulated Transporter1 metal
transporter requires lysine residues. Plant Physiol. 146, 1964e1973.
Khade, H.W., Adholeya, A., 2009. Arbuscular mycorrhizal association in plants
growing on metal-contaminated and noncontaminated soils adjoining Kanpur
tanneries, Uttar Pradesh, India. Water Air Soil Poll. 202, 45e56.
Khan, F.I., Husain, T., Hejazi, R., 2004. An overview and analysis of site remediation
technologies. J. Environ. Manag. 71, 95e122.
Kim, D., Gustin, J.L., Lahner, B., Persans, M.W., Baek, D., Yun, D.J., Salt, D.E., 2004. The
plant CDF family member TgMTP1 from the Ni/Zn hyperaccumulator Thlaspi
goesingense acts to enhance efux of Zn at the plasma membrane when
expressed in Saccharomyces cerevisiae. Plant J. 39, 237e251.
Korenkov, V., Hirschi, K., Crutcheld, J.D., Wagner, G.J., 2007. Enhancing tonoplast
Cd/H antiporter activity increases Cd, Zn and Mn tolerance, and impacts root/
shoot Cd partitioning in Nicotiana tabacum L. Planta 226, 1379e1387.
Kotrba, P., Macek, T., Ruml, T., 1999. Heavy metal-binding peptides and proteins in
plants. Collect. Czech Chem. Commun. 64, 1057e1086.
Kramer, U., Chardonnens, A.N., 2001. The use of transgenic plants in the bioreme-
diation of soils contaminated with trace elements. Appl. Microbiol. Biotechnol.
55, 661e672.
Kramer, U., Pickering, I.J., Prince, R.C., Raskin, I., Salt, D.E., 2000. Subcellular locali-
zation and speciation of Ni in hyperaccumulator and non-accumulator Thlaspi
species. Plant Physiol. 122, 1343e1353.
Kramer, U., Talke, I., Hanikenne, M., 2007. Transition metal transport. FEBS Lett. 581,
2263e2272.
Kramer, U., 2010. Metal hyperaccumulation in plants. Annu. Rev. Plant Biol. 61,
517e534.
Kubota, H., Takenaka, C., 2003. Arabis gemmifera is a hyperaccumulator of Cd and Zn.
Int. J. Phytoremed. 5, 197e201.
Kuklinsky-Sobral, J., Arajo, W.L., Mendes, R., Geraldi, I.O., Pizzirani-Kleiner, A.A.,
Azevedo, J.L., 2004. Isolation and characterization of soybean-associated
bacteria and their potential for plant growth promotion. Environ. Microbiol.
6, 1244e1251.
Kupper, H., Kochian, L.V., 2010. Transcriptional regulation of metal transport genes
and mineral nutrition during acclimatization to cadmium and zinc in the Cd/Zn
hyperaccumulator, Thlaspi caerulescens (Ganges population). New Phytol. 185,
114e129.
Kpper, H., Mijovilovich, A., Gtz, B., Kroneck, P.M.H., Kpper, F.C., Meyer-
Klaucke, W., 2009. Complexation and toxicity of copper in higher plants (I):
characterisation of copper accumulation, speciation and toxicity in Crassula
helmsii as a new copper hyperaccumulator. Plant Physiol. 151, 702e714.
Lasat, M.M., Fuhrmann, M., Ebbs, S.D., Cornish, J.E., Kochian, L.V., 1998. Phytor-
emediation of a radiocesium-contaminated soil: evaluation of cesium-137
bioaccumulation in the shoots of three plant species. J. Environ. Qual. 27,
165e169.
Lasat, M.M., 2000. Phytoextraction of metals from contaminated soil: a review of
plant/soil/metal interaction and assessment of pertinent agronomic issues.
J. Hazard. Subs. Res. 2, 1e25.
Lasat, M.M., 2002. Phytoextraction of toxic metals: a review of biological mecha-
nisms. J. Environ. Qual. 31, 109e120.
Leblanc, M., Petit, D., Deram, A., Robinson, B., Brooks, R.R., 1999. The phytomining
and environmental signicance of hyperaccumulation of thallium by Iberis
intermedia from southern France. Econ. Geol. 94, 109e113.
Lee, S., Moon, J.S., Ko, T.S., Petros, D., Goldsbrough, P.B., Korban, S.S., 2003. Over-
expression of Arabidopsis phytochelatin synthase paradoxycally leads to
hypersensitivity to Cd stress. Plant Physiol. 131, 656e663.
Lee, S., Kim, Y.-Y., Lee, Y., An, G., 2007. Rice P1B-type heavy-metal ATPase, OsHMA9,
is a metal efux protein. Plant Physiol. 145, 831e842.
Li, Y.M., Chaney, R.L., Schneiter, A.A., Miller, J.F., Elias, E.M., Hammond, J.J., 1997.
Screening for low grain Cd genotypes in sunower, durum wheat and ax.
Euphytica 94, 23e30.
Li, Y.M., Chaney, R.L., Brewer, E., Roseberg, R., Angle, J.S., Baker, A., Reeves, R.,
Nelkin, J., 2003. Development of a technology for commercial phytoex-
traction of nickel: economic and technical considerations. Plant Soil 249,
107e115.
Li, Y., Dhankher, O.P., Carreira, L., Balish, R.S., Meagher, R.B., 2005. Arsenic and Hg
tolerance and Cd sensitivity in Arabidopsis plants expressing bacterial gamma-
glutamylcysteine synthetase. Environ. Toxicol. Chem. 24, 1376e1386.
Lin, Z.-Q., Schemenauer, R.S., Cervinka, V., Zayed, A., Lee, A., Terry, N., 2000.
Selenium volatilization from the soil e Salicornia bigelovii Torr. treatment
system for the remediation of contaminated water and soil in the San Joaquin
Valley. J. Environ. Qual. 29, 1048e1056.
Linacre, N.A., Whiting, S.N., Baker, A.J.M., Angle, J.S., Ades, P.K., 2003. Transgenics
and phytoremediation: the need for an integrated risk strategy assessment,
management and communication strategy. J. Phytoremed. 5, 181e185.
Liu, J., Zhu, Q., Zhang, Z., Xu, J., Yang, J., Wong, M.H., 2005. Variations in Cd accu-
mulation among rice cultivars and types and the selection of cultivars for
reducing Cd in the diet. J. Sci. Food Agric. 85, 147e153.
Lopez-Millan, A.F., Ellis, D.R., Grusak, M.A., 2004. Identication and characterization
of several new members of the ZIP family of metal ion transporters in Medicago
truncatula. Plant Mol. Biol. 54, 583e596.
Lugon-Moulin, N., Zhang, M., Gadani, F., Rossi, L., Koller, D., Krauss, M., Wagner, G.J.,
2004. Critical review of the science and options for reducing Cd in tobacco
(Nicotiana tabacum L.) and other plants. Adv. Agron. 83, 111e180.
Luo, S., Wan, Y., Xiao, X., Guo, H., Chen, L., Xi, Q., Zeng, G., Liu, C., Chen, J., 2011.
Isolation and characterization of endophytic bacterium LRE07 from cadmium
hyperaccumulator Solanum nigrum L. and its potential for remediation. Appl.
Microbiol. Biotechnol. 89, 1637e1644.
Lytle, C.M., Lytle, F.W., Yang, N., Qian, J.-H., Hansen, D., Zayed, A., Terry, N., 1998.
Reduction of Cr (VI) to Cr (III) by wetland plants: potential for in situ heavy
metal detoxication. Environ. Sci. Technol. 32, 3087e3093.
Lyyra, S., Meagher, R.B., Kim, T., Heaton, A., Montello, P., Balish, R.S., Merkle, S.A.,
2007. Coupling two Hg resistance genes in Eastern cottonwood enhances the
processing of organomercury. Plant Biotech. J. 5, 254e262.
Ma, L.Q., Komar, K.M., Tu, C., Zhang, W., Cai, Y., Kenelley, E.D., 2001. A fern that
hyperaccumulates arsenic. Nature 409, 579.
Macek, T., Mackov, M., Truksa, M., Singh-Cundy, A., Kotrba, P., Yancey, N.,
Scouten, W.H., 1996. Preparation of transgenic tobacco with a yeast metal-
lothionein combined with a polyhistidine tail. Chem. Listy 90, 690.
Macek, T., Kotrba, P., Svatos, A., Novakova, M., Demnerova, K., Mackov, M., 2007.
Novel roles for genetically modied plants in environmental protection. Trends
Biotech. 26, 146e152.
Macnair, M.R., Bert, V., Huitson, S.B., Saumitou-Laprade, P., Petit, D., 1999. Zn
tolerance and hyperaccumulation are genetically independent characters. Proc.
Royal Soc. Biol. Sci. B 266, 2175e2179.
Macnair, M.R., Tilstone, G.H., Smith, S.S., 2000. The genetics of metal tolerance and
accumulation in higher plants. In: Terry, N., Banuelos, G. (Eds.), Phytor-
emediation of Contaminated Soil and Water. Lewis Publishers, Boca Raton, FL,
pp. 235e250.
Macnair, M.R., 1993. The genetics of metal tolerance in vascular plants. New Phytol.
124, 541e559.
Maestri, E., Marmiroli, M., Visioli, G., Marmiroli, N., 2010. Metal tolerance and
hyperaccumulation: costs and trade-offs between trade and environment. Env.
Exp. Bot. 68, 1e13.
Marques, A.P.G.C., Rangel, A.O.S.S., Castro, P.M.L., 2009. Remediation of heavy metal
contaminated soils: phytoremediation as a potentially promising clean-up
technology. Crit. Rev. Environ. Sci. Tech. 39, 622e654.
McGrath, S.P., Zhao, F.J., 2003. Phytoextraction of metals and metalloids from
contaminated soils. Curr. Opin. Biotechnol. 14, 277e282.
McGrath, S.P., Zhao, F.J., Lombi, E., 2002. Phytoremediation of metals, metalloids and
radionuclides. Adv. Agron. 75, 1e56.
McGrath, S.P., Lombi, E., Gray, C.W., Caille, N., Dunham, S.J., Zhao, F.J., 2006.
Field evaluation of Cd and Zn phytoextraction potential by the hyper-
accumulators Thlaspi caerulescens and Arabidopsis halleri. Environ. Poll. 141,
115e125.
McLaughlin, M.J., Parker, D.R., Clarke, J.M., 1999. Metals and micronutrients e food
safety issues. Field Crops Res. 60, 143e163.
Meagher, R.B., Rugh, C.L., Kandasamy, M.K., Gragson, G., Wang, N.J., 2000. Engi-
neered phytoremediation of Hg pollution in soil and water using bacterial
genes. In: Terry, N., Banuelos, G. (Eds.), Phytoremediation of Contaminated Soil
and Water. Lewis Publishers, Boca Raton, FL, pp. 201e221.
Meers, E., Ruttens, A., Hopgood, M., Lesage, E., Tack, F.M.G., 2005. Potential of
Brassica rapa, Cannabis sativa, Helianthus annuus, and Zea mays for phytoex-
traction of heavy metals from calcareous dredged sediment derived soils.
Meers, E., Vandecasteele, B., Ruttens, A., Vangronsveld, J., Tack, F.M.G., 2007.
Potential of ve willow species (Salix spp.) for phytoextraction of heavy metals.
Environ. Exp. Bot. 60, 57e68.
Memon, A.R., Schroder, P., 2009. Implications of metal accumulation mechanisms to
phytoremediation. Environ. Sci. Poll. Res. 16, 162e175.
Mench, M., Lepp, N., Bert, V., Schwitzgubel, J.P., Gawronski, S.W., Schrder, P.,
Vangronsveld, J., 2010. Successes and limitations of phytotechnologies at eld
scale: outcome, assessment and outlook from COST Action 859. J. Soil Sedim 10,
1039e1070.
Mertens, J., Luyssaert, S., Verheyen, K., 2005. Use and abuse of trace metal
concentrations in plant tissue for biomonitoring and phytoextraction. Environ.
Poll. 138, 1e4.
Mertens, J., Vervaeke, P., Meers, E., Tack, F.M.G., 2006. Seasonal changes of metals in
willow (Salix sp.) stands for phytoremediation on dredged sediment. Environ.
Sci. Technol. 40, 1962e1968.
Mertens, J., van Nevel, L., De Schrijver, A., Piesschaert, F., Oosterbean, A., Tack, F.M.G.,
Verheyen, K., 2007. Tree species effect on the redistribution of soil metals.
Environ. Poll. 149, 173e181.
Miransari, M., 2011. Hyperaccumulators, arbuscular mycorrhizal fungi and stress of
heavy metals. Biotechnol. Adv. 29, 645e653.
Mizuno, T., Usui, K., Horie, K., Nosaka, S., Mizuno, N., Obata, H., 2005. Cloning of
three ZIP/NRAMP transporter genes from a Ni hyperaccumulator plant Thlaspi
japonicum and their Ni 2-transport abilities. Plant Physiol. Biochem. 43,
793e801.
Moffat, A.S., 1999. Engineering plants to cope with metals. Science 285, 369e370.
Mongkhonsin, B., Nakbanpote, W., Nakai, I., Hokura, A., Jearanaikoon, N., 2011.
Distribution and speciation of chromium accumulated in Gynura pseudochina
(L.) DC. Environ. Exp. Bot. 74, 56e64.
Moradi, A.B., Swoboda, S., Robinson, B., Prohaska, T., Kaestner, A., Oswald, S.E.,
Wenzel, W.W., Schulin, R., 2010. Mapping of nickel in root cross-sections of the
hyperaccumulator plant Berkheya coddii using laser ablation ICP-MS. Environ.
Exp. Bot. 69, 24e31.
Morel, M., Crouzet, J., Gravot, A., Auroy, P., Leonhardt, N., Vavasseur, A., Richaud, P.,
2008. AtHMA3, a P
1B
-ATPase allowing Cd/Zn/Co/Pb vacuolar storage in Arabi-
dopsis. Plant Physiol. 149, 894e904.
Na, G., Salt, D.E., 2011. Differential regulation of serine acetyltransferase is involved
in nickel hyperaccumulation in Thlaspi goesingense. J. Biol. Chem. 286,
40423e40432.
Nagata, T., Ishikawa, C., Kiyono, M., Pan-Hou, H., 2006. Accumulation of Hg in
transgenic tobacco expressing bacterial polyphosphate. Biol. Pharm. Bull. 29,
2350e2353.
Nair, A., Juwarkar, A.A., Singh, S.K., 2007. Production and characterization of side-
rophores and its application in arsenic removal from contaminated soil. Water
Air Soil Poll. 180, 199e212.
Nanda-Kumar, P.B.A., Dushenkov, V., Motto, H., Raskin, I., 1995. Phytoextraction: the
use of plants to remove heavy metals from soil. Environ. Sci. Tech. 29,
1232e1238.
Narwal, R.P., Singh, B.R., 1998. Effect of organic materials on partitioning, extract-
ability and plant uptake of metals in an alum shale soil. Water Air Soil Poll. 103,
405e421.
Nedelkoska, T.V., Doran, P.M., 2000. Hyperaccumulation of Cd by hairy roots of
Thlaspi caerulescens. Biotechnol. Bioeng. 67, 607e615.
Nehnevajova, E., Herzig, R., Erismann, K.-H., Schwitzgubel, J.-P., 2007. In vitro
breeding of Brassica juncea L. to enhance metal accumulation and extraction
properties. Plant Cell Rep. 26, 429e437.
Neubauer, U., Furrer, G., Kayser, A., Schulin, R., 2000. Siderophores, NTA, and citrate:
potential soil amendments to enhance heavy metal mobility in phytor-
emediation. Int. J. Phytoremed. 2, 353e368.
Nie, L., Shah, S., Rashid, A., Burd, G.I., Dixon, D.G., Glick, B.R., 2002. Phytoremediation
of arsenate contaminated soil by transgenic canola and the plant growth-
promoting bacterium Enterobacter cloacae CAL2. Plant Physiol. Biochem. 40,
355e361.
Nikolic, D.B., Samardzic, J.T., Bratic, A.M., Radin, I.P., Gavrilovic, S.P., Rausch, T.,
Maksimovic, V.R., 2010. Buckwheat (Fagopyrum esculentum Moench) FeMT3
gene in heavy metal stress: protective role of the protein and inducibility of
the promoter region under Cu
2
and Cd
2
treatments. J. Agric. Food Chem. 58,
3488e3494.
Noret, N., Meerts, P., Vanhaelen, M., Dos Santos, A., Escarr, J., 2007. Do metal-rich
plants deter herbivores? A eld test of the defence hypothesis. Oecologia 152,
92e100.
Oven, M., Grill, E., Golan-Goldhirsh, A., Kutchan, T.M., Zenk, M.H., 2002. Increase of
free cysteine and citric acid in plant cells exposed to cobalt ions. Phytochem-
istry 60, 467e474.
Padmavathiamma, P.K., Li, L.Y., 2007. Phytoremediation technology: hyper-
accumulation metals in plants. Plant Soil 184, 105e126.
Pan, A.H., Yang, M., Tie, F., Li, L., Che, Z., Ru, B., 1994. Expression of mouse
metallothionein-I gene confers Cd resistance in transgenic tobacco plants. Plant
Mol. Biol. 24, 341e351.
Parker, D.R., Chaney, R.L., Norvell, W.A., 1995. Equilibrium computer models:
applications to plant nutrition research. In: Loeppert, R.H. (Ed.), Chemical
Equilibrium and Reaction Models. SSSA Special Publ. No. 42. ASA and SSSA,
Madison, WI, pp. 163e200.
Perrier, N., 2004. Nickel speciation in Sebertia acuminata, a plant growing on
a lateritic soil of New Caledonia. Geoscience 336, 567e577.
Perronnet, K., Schwartz, C., Gerard, E., Morel, J.L., 2000. Availability of Cd and Zn
accumulated in the leaves of Thlaspi caerulescens incorporated into soil. Plant
Soil 227, 257e263.
Persans, M.W., Yan, X., Patnoe, J.-M.M.L., Kramer, U., Salt, D.E., 1999. Molecular
dissection of the role of histidine in Ni hyperaccumulation in Thlaspi
goesingense (Halacsy). Plant Physiol. 121, 1117e1126.
Persans, M.W., Nieman, K., Salt, D.E., 2001. Functional activity and role of cation-
efux family members in Ni hyperaccumulation in Thlaspi goesingense. Proc.
Natl. Acad. Sci. (USA) 98, 9995e10000.
Peuke, A.D., Rennenberg, H., 2005. Phytoremediation: molecular biology, require-
ments for application, environmental protection, public attention and feasi-
bility. EMBO Rep. 6, 497e501.
Pianelli, K., Mari, S., Marqus, L., Lebrun, M., Czernic, P., 2005. Nicotianamine
over-accumulation confers resistance to nickel in Arabidopsis thaliana.
Transgenic Res. 14, 739e748.
Pickering, I.J., Prince, R.C., George, M.J., Smith, R.D., George, G.N., Salt, D.E., 2000.
Reduction and coordination of arsenic in Indian mustard. Plant Physiol. 122,
1171e1177.
Pilon-Smits, E.A.H., Pilon, M., 2000. Breeding Hg-breathing plants for environ-
mental cleanup. Trends Plant Sci. 5, 235e236.
Pilon-Smits, E.A.H., Pilon, M., 2002. Phytoremediation of metals using transgenic
plants. Crit. Rev. Plant Sci. 21, 439e456.
Pilon-Smits, E.A.H., Hwang, S., Lytle, C., Zhu, Y., Tai, J.C., Bravo, R.C., Chen, Y.,
Leustek, T., Terry, N., 1999. Overexpression of ATP sulfurylase in Indian mustard
Pbs to increased selenate uptake, reduction, and tolerance. Plant Physiol. 119,
123e132.
Pinel, F., Leclerc-Cessac, E., Staunton, S., 2003. Relative contributions of soil chem-
istry, plant physiology, and rhizosphere-induced changes in speciation on Ni
accumulation in plant shoots. Plant Soil 255, 619e629.
Plaza, S., Tearall, K.L., Zhao, F.J., Buchner, P., McGrath, S.P., Hawkesford, M.J., 2007.
Expression and functional analysis of metal transporter genes in two con-
trasting ecotypes of the hyperaccumulator Thlaspi caerulescens. J. Exp. Bot. 58,
1717e1728.
Pollard, A.J., Stewart, H.L., Roberson, C.B., 2009. Manganese hyperaccumulation in
Phytolacca americana L. from the southeastern united states. Northeastern
Natur. 16, 155e162.
Prasad, M.N.V., Freitas, H., 2003. Metal hyperaccumulation in plants e biodiversity
prospecting for phytoremediation technology. Electronic J. Biotechnol. 6,
275e321.
Prasad, M.N.V., 2003. Phytoremediation of metal polluted ecosystems: hype for
commercialization. Russian J. Plant Physiol. 50, 686e700.
Pulford, I.D., Watson, C., 2003. Phytoremediation of heavy metal-contaminated land
by trees e a review. Environ. Int. 29, 529e540.
Puschenreiter, M., Wieczorek, S., Horak, O., Wenzel, W.W., 2003. Chemical changes
in the rhizosphere of metal hyperaccumulator and excluder Thlaspi species.
J. Plant Nutr. Soil Sci. 166, 579e584.
Rajkumar, M., Prasad, M.N.V., Freitas, H., Ae, N., 2009. Biotechnological applications
of serpentine bacteria for phytoremediation of heavy metals. Crit. Rev. Biotech.
29, 120e130.
Rascio, N., Navari-Izzo, F., 2011. Heavy metal accumulating plants: how and why do
they do it? And what makes them so interesting? Plant Sci. 180, 169e181.
Raskin, I., Smith, R.D., Salt, D.E., 1997. Phytoremediation of metals: using
plants to remove pollutants from the environment. Curr. Opin. Biotechnol. 8,
221e226.
Raskin, I., 1996. Plant genetic engineering may help with environmental cleanup.
Proc. Natl. Acad. Sci. (USA) 93, 3164e3166.
Reeves, R.D., Baker, A.J.M., 2000. Metal accumulating plants. In: Raskin, I.,
Ensley, B.D. (Eds.), Phytoremediation of Toxic Metals: Using Plants to Clean Up
the Environment. Wiley, New York, pp. 193e229.
Reeves, R.D., Baker, A.J.M., Borhidi, A., Berazain, R., 1999. Nickel hyperaccumulation
in the serpentine ora of Cuba. Ann. Bot. 83, 29e38.
Reeves, R.D., Adiguzel, N., Baker, A.J.M., 2009. Nickel hyperaccumulation in
Bornmuellera kiyakii and associated plants of the Brassicaceae from Kzlda
Derebucak (Konya), Turkey. Turk. J. Bot. 33, 33e40.
Reisinger, S., Schiavon, M., Terry, N., Pilon-Smits, E.A., 2008. Heavy metal tolerance
and accumulation in Indian mustard (Brassica juncea L.) expressing bacterial
gamma-glutamylcysteine synthetase or glutathione synthetase. Int. J. Phytor-
emed. 10, 440e454.
Richau, K.H., Schat, H., 2009. Intraspecic variation of Ni and Zn accumulation
and tolerance in the hyperaccumulator Thlaspi caerulescens. Plant Soil 314,
253e262.
Robinson, B.H., Brooks, R.R., Howes, A.W., Kirkman, J.H., Gregg, P.E.H., 1997. The
potential of the high-biomass nickel hyperaccumulator Berkheya coddii for
phytoremediation and phytomining. J. Geochem. Explor. 60, 115e126.
Robinson, B., Fernandez, J.E., Madejon, P., Maranon, T., Murillo, J.M., Green, S.,
Clotheir, B., 2003. Phytoextraction: an assessment of biogeochemical and
economic viability. Plant Soil 249, 117e125.
Robinson, B.H., Schulin, R., Nowack, B., Roulier, S., Menon, M., Clothier, B., Green, S.,
Mills, T., 2006. Phytoremediation for the management of metal ux in
contaminated sites. Forest Snow Landscape Res. 80, 221e234.
Roosens, N.H., Willems, G., Saumitou-Laprade, P., 2008a. Using Arabidopsis to
explore zinc tolerance and hyperaccumulation. Trends Plant Sci. 13, 208e215.
Roosens, N.H.C.J., Glenda, W., Ccile, G., Adeline, C., Pierre, S.L., 2008b. The use of
comparative genome analysis and synthetic relationships allows extrapolating
the position of Zn tolerance QTL regions from Arabidopsis halleri into Arabidopsis
thaliana. Plant Soil 306, 105e116.
Rosselli, W., Keller, C., Boschi, K., 2003. Phytoextraction capacity of trees growing on
a metal contaminated soil. Plant Soil 256, 265e272.
Rugh, C.L., Wilde, H.D., Stack, N.M., Thompson, D.M., Summers, A.O., Meagher, R.B.,
1996. Mercuric ion reduction and resistance in transgenic Arabidopsis thaliana
plants expressing a modied bacterial merA gene. Proc. Natl. Acad. Sci. (USA) 93,
3182e3187.
Rugh, C.L., Senecoff, J.F., Meagher, R.B., Merkle, S.A., 1998. Development of trans-
genic yellow poplar for Hg phytoremediation. Nat. Biotech. 16, 925e928.
Rugh, C.L., Bizily, S.P., Meagher, R.B., 2000. Phytoreduction of environmental
Hg pollution. In: Raskin, I., Ensley, B.D. (Eds.), Phytoremediation of Toxic
Metals e Using Plants to Clean Up the Environment. Wiley, New York,
pp. 151e171.
Ryan, P.R., Delhaize, E., Jones, D.L., 2001. Function and mechanism of organic
anion exudation from plant roots. Ann. Rev. Plant Physiol. Plant Mol. Biol. 5,
527e560.
Sahi, S.V., Bryant, N.L., Sharma, N.C., Singh, S.R., 2002. Characterization of a lead
hyperaccumulator shrub, Sesbania drummondii. Environ. Sci. Technol. 36,
4676e4680.
Saikkonen, K., Lehtonen, P., Helander, M., Koricheva, J., Faeth, S.H., 2006. Model
systems in ecology: dissecting the endophyteegrass literature. Trends Plant Sci.
11, 428e433.
Sancenon, V., Puig, S., Mateu-Andres, I., Dorcey, E., Thiele, D.J., Penarrubia, L., 2004.
The Arabidopsis copper transporter COPT1 functions in root elongation and
pollen development. J. Biol. Chem. 279, 15348e15355.
Sasaki, T., Yamamoto, Y., Ezaki, B., Katsuhara, M., Ahn, S.J., Ryan, P.R., Delhaize, E.,
Matsumoto, H., 2004. A wheat gene encoding an aluminum-activated malate
transporter. Plant J. 37, 645e653.
Sas-Nowosielska, A., Kucharski, R., Pogrzeba, M., KrzyZak, J., Kuperberg, J.M.,
Japenga, J., 2008. Phytoremediation technologies used to reduce environmental
threat posed by metal contaminated soils: theory and reality. In: Barnes, I.,
Kharytonov, M.M. (Eds.), Simulation and Assessment of Chemical Processes in
a Multiphase Environment. Springer, New York, pp. 285e297.
Schaaf, G., Honsbein, A., Meda, A.R., Kirchner, S., Wlpf, D., von Viren, N., 2006.
AtIREG2 encodes a tonoplast transport protein involved in iron-dependent
nickel detoxication in Arabidopsis thaliana roots. J. Biol. Chem. 281,
25532e25540.
Schikora, A., Thimm, O., Linke, B., Buckhout, T.J., Mller, M., Schmidt, W., 2006.
Expression, localization, and regulation of the iron transporter LeIRT1 in tomato
roots. Plant Soil 284, 101e108.
Schmidt, U., 2003. Enhancing phytoextraction: the effect of chemical soil manipu-
lation on mobility, plant accumulation and leaching of heavy metals. J. Environ.
Qual. 32, 1939e1954.
Schmoger, M.E.V., Oven, M., Grill, E., 2000. Detoxication of arsenic by phytoche-
latins in plants. Plant Physiol. 122, 793e801.
Sekhar, K., Priyanka, B., Reddy, V.D., Rao, K.V., 2011. Metallothionein 1 (CcMT1) of
pigeonpea (Cajanus cajan L.) confers enhanced tolerance to copper and
cadmium in Escherichia coli and Arabidopsis thaliana. Environ. Exp. Bot..
doi:10.1016/j.envexpbot.2011.02.017
Seth, C.S., 2011. A review on mechanisms of plant tolerance and role of transgenic
plants in environmental clean-up. Bot. Rev. (Online DOI 10.1007/s12229-011-
9092-x).
Shah, K., Nongkynrih, J.M., 2007. Metal hyperaccumulation and bioremediation.
Biol. Plant 51, 618e634.
Sharma, N.C., Gardea-Torresday, J.L., Parson Sahi, S.V., 2004. Chemical speciation of
lead in Sesbania drummondii. Environ. Toxicol. Chem. 23, 2068e2073.
Sheng, J., Liu, K., Fan, B., Yuan, Y., Shen, L., Ru, B., 2007. Improving zinc content and
antioxidant activity in transgenic tomato plants with expression of mouse
metallothionein-I by mt-1 gene. J. Agric. Food Chem. 55, 9846e9849.
Shingu, Y., Kudo, T., Ohsato, S., Kimura, M., Ono, Y., Yamaguchi, I., Hamamoto, H.,
2005. Characterization of genes encoding metal tolerance proteins isolated
from Nicotiana glauca and Nicotiana tabacum. Biochem. Biophys. Res. Comm.
331, 675e680.
Singh, O.V., Labana, S., Pandey, G., Budhiraja, R., Jain, R.K., 2003. Phytoremediation:
an overview of metallic ion decontamination from soil. Appl. Microbiol.
Biotechnol. 61, 405e412.
Singla-Pareek, S.L., Yadav, S.K., Pareek, A., Reddy, M.K., Sopory, S.K., 2006. Transgenic
tobacco overexpressing glyoxalase pathway enzymes grow and set viable seeds
in Zn-spiked soils. Plant Physiol. 140, 613e623.
Smith, S.E., Read, D.J., 2008. Mycorrhizal Symbiosis. Academic Press, San Diego.
Song, W.Y., Sohn, E.J., Martinoia, E., Lee, Y.J., Yang, Y.Y., Jasinski, M., Forestier, C.,
Hwang, I., Lee, Y., 2003. Engineering tolerance and accumulation of Pb and Cd in
transgenic plants. Nat. Biotech. 21, 914e919.
Stearns, J.C., Shah, S., Greenberg, B.M., Dixon, D.G., Glick, B.R., 2005. Tolerance of
transgenic canola expressing 1-aminocyclopropane-1-carboxylic acid deami-
nase to growth inhibition by Ni. Plant Physiol. Biochem. 43, 701e708.
Steele, M.C., Pichtel, J., 1998. Ex-situ remediation of metal contaminated superfund
soil using selective extractants. J. Environ. Eng. 124, 639e645.
Stomp, A.M., Han, K.L., Wilbert, S., Gordon, M.P., 1993. Genetic improvement of
tree species for remediation of hazardous wastes. In Vitro Cell. Dev. Biol. 29,
227e232.
Sun, Q., Ye, Z.H., Wang, X.R., Wong, M.H., 2005. Increase of glutathione in mine
population of Sedum alfredii: a Zn hyperaccumulator and Pb accumulator.
Phytochemistry 66, 2549e2556.
Sun, Y., Zhou, Q., Wang, L., Liu, W., 2009. Cadmium tolerance and accumulation
characteristics of Bidens pilosa L. as a potential Cd-hyperaccumulator.
J. Hazardous Mat. 161, 808e814.
Szira, F., Balint, A.F., Vagujfalvi, A., Galiba, G., 2008. Are Cbf genes involved in copper
tolerance? Acta Biologica Szegediensis 52, 205e207.
Taghavi, S., van der Lelie, D., Hoffman, A., Zhang, Y.-B., Walla, M.D., Vangronsveld, J.,
Newman, L., Monchy, S., 2010. Genome sequence of the plant growth promoting
endophytic bacterium Enterobacter sp. 638. PLoS Genet. 6, e1000943.
Talke, I., Hanikenne, M., Krmer, U., 2006. Zn dependent global transcriptional
control, transcriptional de-regulation and higher gene copy number for genes
in metal homeostasis of the hyperaccumulator Arabidopsis halleri. Plant Physiol.
142, 148e167.
Tamura, H., Honda, M., Sato, T., Kamachi, H., 2005. Pb hyperaccumulation and
tolerance in commom buckwheat (Fagopyrum esculentum Moench). J. Plant Res.
118, 355e359.
Tang, Y.-T., Qiu, R.-L., Zeng, X.-W., Ying, R.-R., Yu, F.-M., Zhou, X.-Y., 2009. Lead, zinc
cadmium accumulation and growth simulation in Arabis paniculata Franch. Env.
Exp. Bot. 66, 126e134.
Thangavel, P., Subhuram, C.V., 2004. Phytoextraction e role of hyper accumulators
in metal contaminated soils. Proc. Indian Natl. Sci. Acad. Part B 70, 109e130.
Thomas, J.C., Davies, E.C., Malick, F.K., Endreszl, C., Williams, C.R., Abbas, M.,
Petrella, S., Swisher, K., Perron, M., Edwards, R., Ostenkowski, P., Urbanczyk, N.,
Wiesend, W.N., Murray, K.S., 2003. Yeast metallothionein in transgenic tobacco
promotes Cu uptake from contaminated soils. Biotech. Prog. 19, 273e280.
Tiller, K.G., Oliver, D.P., Mclaughlin, M.J., Merry, R.H., Naidu, R., 1995. Managing Cd
contamination of agricultural land. In: Adriano, D.C. (Ed.), Biogeochemistry of
Trace Elements-2. Advances in Environmental Science. Science Reviews
Limited, Northwood, U.K.
Tong, Y.P., Kneer, R., Zhu, Y.G., 2004. Vacuolar compartmentalization: a second-
generation approach to engineering plants for phytoremediation. Trends
Plant Sci. 9, 7e9.
Turchi, A., Tamantini, I., Camussi, A.M., Racchi, M.L., 2012. Expression of a metal-
lothionein A1 gene of Pisum sativum in white poplar enhances tolerance and
accumulation of zinc and copper. Plant Sci. 183, 50e56.
Unterbrunner, R., Puschenreiter, M., Sommer, P., Wieshammer, G., Tlustos, P.,
Zupan, M., 2007. Heavy metal accumulation in tree growing on contaminated
sites in Central Europe. Environ. Poll. 148, 107e114.
Vamerali, T., Bandiera, M., Mosca, G., 2010. Field crops for phytoremediation of
metal-contaminated land. A review. Environ. Chem. Lett. 8, 1e17.
van de Mortel, J.E., Villanueva, L.A., Schat, H., Kwekkeboom, J., Coughlan, S.,
Moerland, P.D., Ver Loren van Themaat, E., Koornneef, M., Aarts, M.G.M., 2006.
Large expression differences in genes for iron and Zn homeostasis, stress
response, and lignin biosynthesis distinguish roots of Arabidopsis thaliana and
the related metal hyperaccumulator Thlaspi caerulescens. Plant Physiol. 142,
1127e1147.
van Hoof, N.A.L.M., Hassinen, V.H., Hakvoort, H.W.J., Ballintijn, K.F., Schat, H.,
Verkleij, J.A.C., Ernst, W.H.O., Karenlampi, S.O., Tervahauta, A.I., 2001. Enhanced
copper tolerance in Silene vulgaris (Moench) Garcke populations from copper
mines is associated with increased transcript levels of a 2b-type metal-
lothionein gene. Plant Physiol. 126, 1519e1526.
van Huysen, T., Abdel-Ghany, S., Hale, K.L., LeDuc, D., Terry, N., Pilon-Smits, E.A.H.,
2003. Overexpression of cystathionine-g-synthase enhances selenium volatili-
zation in Brassica juncea. Planta 218, 71e78.
van Nevel, L.V., Mertens, J., Oorts, K., Verheyen, K., 2007. Phytoextraction of metals
from soil e how far from practice? Environ. Poll. 150, 34e40.
Verbruggen, N., Hermans, C., Schat, H., 2009. Molecular mechanisms of metal
hyperaccumulation in plants. New Phytol. 181, 759e776.
Wang, H., Zhong, G., 2011. Effect of organic ligands on accumulation of copper in
hyperaccumulator and nonaccumulator Commelina communis. Biol. Trace Elem.
Res. 143, 489e499.
Wang, A.S., Angle, J.S., Chaney, R.L., Delorme, T.A., Reeves, R.D., 2006. Soil pH effects
on uptake of Cd and Zn by Thlaspi caerulescens. Plant Soil 281, 325e337.
Wangeline, A.L., Burkhead, J.L., Hale, K.L., Lindblom, S.D., Terry, N., Pilon, M.,
Pilon-Smits, E.A.H., 2004. Overexpression of ATP sulfurylase in Indian mustard:
effects on tolerance and accumulation of twelve metals. J. Environ. Qual. 33,
54e60.
Watrud, L.S., Misra, S., Gidamu, L., Shiroyama, T., Maggard, S., Giovanni, G.D., 2006.
Ecological risk assessment of alfalfa (Medicago varia L.) genetically engineered
to express a human metallothionein (hMT) gene. Water Air Soil Poll. 176,
329e349.
Weber, M., Harada, E., Vess, C., von Roepenack-Lahaye, E., Clemens, S., 2004.
Comparative microarray analysis of Arabidopsis thaliana and Arabidopsis halleri
roots identies nicotianamine synthase, a ZIP transporter and other genes as
potential metal hyperaccumulation factors. Plant J. 37, 269e281.
Wenzel, W.W., Bunkowski, M., Puschenreiter, M., Horak, O., 2003a. Rhizosphere
characteristics of indigenously growing nickel hyperaccumulator and excluder
plants on serpentine soil. Environ. Poll. 123, 131e138.
Wenzel, W.W., Unterbrunner, R., Sommer, P., Sacco, P., 2003b. Chelate-assisted
phytoextraction using canola (Brassica napus L.) in outdoors pot and lysimeter
experiments. Plant Soil 249, 83e96.
Willems, G., Drager, D.B., Courbot, M., Gode, C., Verbruggen, N., Saumitou-
Laprade, P., 2007. The genetic basis of Zn tolerance in the metallophyte
Arabidopsis halleri ssp. halleri (Brassicaceae): an analysis of quantitative trait
loci. Genetics 176, 659e674.
Williams, L.E., Pittman, J.K., Hall, J.L., 2000. Emerging mechanisms for heavy metal
transport in plants. Biochim. Biophys. Acta 1465, 104e126.
Wong, S.C., Li, X.D., Zhang, G., Qi, S.H., Min, Y.H., 2002. Heavy metals in agricultural
soils of the Pearl River Delta, South China. Environ. Poll. 119, 33e44.
Wong, M.H., 2003. Ecological restoration of mine degraded soils, with emphasis on
metal contaminated soils. Chemosphere 50, 775e780.
Wray, G., Babbitt, C.C., 2008. Enhancing gene regulation. Science 321, 1300e1301.
Xiao, H., Yin, L., Xu, X., Li, T., Han, Z., 2008. The Iron-regulated transporter,
MbNRAMP1, isolated from Malus baccata is involved in Fe, Mn and Cd
trafcking. Ann. Bot. 102, 881e889.
Xing, J.P., Jiang, R.F., Ueno, D., Ma, J.F., McGrath, S.P., Zhao, F.J., 2008. Variation in
root-to-shoot translocation of cadmium and zinc among different accessions of
the hyperaccumulators Thlaspi caerulescens and Thlaspi praecox. New Phytol.
178, 315e325.
Xu, J., Tian, Y.S., Peng, R.H., Xiong, A.S., Zhu, B., Jin, X.F., Gao, J.J., Hou, X.L., Yao, Q.H.,
2009. Yeast copper-dependent transcription factor ACE1 enhanced copper
stress tolerance in Arabidopsis. BMB Rep. 42, 752e757.
Xu, J., Tian, Y.S., Peng, R.H., Xiong, A.S., Zhu, B., Hou, X.L., Yao, Q.H., 2010. Cyano-
bacteria MT gene SmtA enhances zinc tolerance in Arabidopsis. Mol. Biol. Rep.
37, 1105e1110.
Yang, Z., Wu, Y., Li, Y., Ling, H.Q., Chu, C., 2009. OsMT1a, a type 1 metallothionein,
plays the pivotal role in zinc homeostasis and drought tolerance in rice. Plant
Mol. Biol. 70, 219e229.
Yehuda, Z., Shenker, M., Rmheld, V., Marschner, H., Hadar, Y., Chen, Y., 1996. The
role of ligand exchange in the uptake of iron from microbial siderophores by
gramineous plants. Plant Physiol. 112, 1273e1280.
Yu, X.Z., Trapp, S., Zhou, P.H., Hu, H., 2005. The effect of temperature on the rates of
cyanide metabolism of two woody plants. Chemosphere 59, 1099e1104.
Yu, X.Z., Peng, X.Y., Xing, L.Q., 2010. Effect of temperature on phytoextraction of
hexavalent and trivalent chromium by hybrid willows. Ecotoxicology 19,
61e68.
Zhang, W., Cai, Y., Tu, C., Ma, L.Q., 2002. Arsenic speciation and distribution in an
arsenic hyperaccumulating plant. Sci. Total Environ. 300, 167e177.
Zhang, Y.W., Tam, N.F.Y., Wong, Y.S., 2004. Cloning and characterization of type 2
metallothionein-like gene from a wetland plant, Typha latifolia. Plant Sci. 167,
869e877.
Zhang, X.H., Liu, J., Huang, H.T., Chen, J., Zhu, Y.N., Wang, D.Q., 2007. Chromium
accumulation by the hyperaccumulator plant Leersia hexandra Swartz.
Zhao, F.J., Lombi, E., Breedon, T., McGrath, S.P., 2000. Zinc hyperaccumulation and
cellular distribution in Arabidopsis halleri. Plant Cell Environ. 23, 507e514.
Zhao, F.J., Hamon, R.E., McLaughlin, M.J., 2001. Root exudates of the hyper-
accumulator Thlaspi caerulescens do not enhance metal mobilization. New
Phytol. 151, 613e620.
Zhu, Y.L., Pilon-Smits, E.A.H., Tarun, A.S., Weber, S.U., Jouanin, L., Terry, N., 1999. Cd
tolerance and accumulation in Indian mustard is enhanced by overexpressing
g-glutamylcysteine synthetase. Plant Physiol. 121, 1169e1177.

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