Phytoremediation : A technologies to remediate soil contaminated

with heavy metals.

V.K. Sinhal1, Alok Srivastava2 & V.P. Singh2
1. Maharaja Agrasen Mahavidyalaya, Bareilly, U.P. India
2. Deptt. of Plant Science, M.J.P. Rohilkhand University, Bareilly U.P. India

Abstract:
It is almost impossible to imagine soil without atleast trace levels of
heavy metals. Natural and anthropogenic activities concentrated some of
these elements in certain areas upto hazardous levels for living organisms.
The non-biodegradable metals accumulate in the environment and
subsequently contaminate the food chain. The elevated level of heavy
metals poses a risk to human health. These heavy metals drastically can alter
the metabolic activities of organism. Heavy metals may be carcinogenic,
mutagenic and teratogenic. Thus phytoremediation of heavy metals deserves
great attention. Most of the conventional remedial technologies are
expensive and may inhibit the soil fertility. In contrast, phytoremediation is
a cost effective, environmentally friendly pleasing approach most suitable
for developing countries. Among several subsets of phytoremediation, the
widely studied strategies are (a) phytoextration, (b) phytofiletreation, (c)
phytovolatilization, (d) phytostabilization, (e) phytodegradation and (f)
rhizodegradation. Now a days new efficient metal hyperaccumulators are
being used in phytoremediation & phytomining. Various techniques to
enhance phytoremediation and utilization of by-products have been
elaborated. Present investigation reports about the mobility, bioavailablity
and plant response to heavy metals and future trends in phytoremediation to
remediate the soil and water contaminated with heavy metals.
Key words: Environmental pollution, Heavy metals, Phytoremediation
Hyperaccumulator
1

1.

Introduction
Anthropogenic activities like mining,

combustion of fossil fuels,

phosphate fertilizers, military activities, metal working industries

and

natural processes such as volcanic eruptions, continental dust etc. lead to

emission of heave metals and accumulation of these chemicals in
environment (Hooda, 2007). It is evident that anthropogenic sources are
responsible for pollution and environmental degradation in order to exploit
nature for means of livelihood. All components of the biosphere are facing
threats of pollution by a variety of organic/inorganic pollution because of
manmade activities that alter the normal biogeochemical cycle (Ali et al.
2013).
Various engineering based methods such as soil excavation, soil
washing or burning or pump and treat systems are already being used to
remediate metal contaminated soils. However, these non biological
techniques are not fully acceptable as they destroy the biotic components of
soil and are technically difficult and expensive to implement (Hooda, 2007).
Now a days phytoremedation is an emerging promising approach to
remediate the polluted environments (Pulford and Watson 2003; Sinhal et al.
2011). This technology is an alternative or complimentary one that could be
applied along with or instead of mechanical congenital cleaning
methodologies which mostly require high capital input, labour and intensive
energy. Certain plants have endogenous, genetic, biochemical and
physiological qualities to combat against the soil, water and air pollution
(Cunningham and Berti 1993; Meagher, 2000). Phytoremediation takes
advantage of uptake capabilities of plant root systems, together with the
translocation, bioaccumulation and pollutant storage/degradation abilities of
the entire plant body.

Heavy metals are essentially nonbiodegradable and therefore

accumulate in the environment. The accumulation of heavy metals in soils
and waters poses a risk to the environment and human health. These heavy
metals accumulate in the body tissues of living organism and shows
phytotoxic, eytogentic & biochemical effects at dangerous level (Sinhal et
al. 2007). On the basic role of heavy metal in biological systems they are
categorized into essential and non-essential. Fe, Mn, Zn, Cu etc. are some
essential metals which are needed by plants in minute quantities whereas
Cd, Pb, As, Hg, Cr, etc. are non-essential heavy metals not needed by living
organism for metabolic activities (Peng et al. 2009; Sanchoz Chardi et al
2009; Dabonne et al. 2010). Heavy metals concentrations beyond threshold
limits have adverse effect on biological system because they can alter or
interfere the normal functioning of living organisms. The present
investigation focuses studies on the phytoremediation technologies
especially phytoextraction and future trends in phytoremediation.
2.

Catastrophic effect of Heavy metals on metabolism of organisms

Different heavy metals differently effect the metabolic activities of

organisms. Cd is of particulars concern to human health because it

accumulates in the body with a half-life exceeding 10 years and has been
linked with renal tubular dysfunction (Buchet et al. 1990), pulmonary
emphysema (Ryan et al; 1982), and possibly osteoporosis (Bhattacharyya et
al. 1988). Heavy metals like Pb, Hg, As, Cr etc. at very low concentration
shows phytotoxic effects in term of reduced levels of protein carbohydrate,
nitrogen, chlorophyll, carotenoids (Sinhal et al. 2010). and also reduction in
activities of enzymes like Nitrate reductare (NR), Glutamate dehydrogenase
(GDH) and Glutamine oxoglutarate amino transferase (GOGAT). Heavy
metals cause oxidative stress (Mudipalli, 2008) by formation of free
redicals. Oxidative stress refers to enhanced generation of reactive oxygen
3

species (ROS) which can overwhelm cells intrinsic antioxidant defenses

and lead to cell damage or death (Das et al 2008; Krystofova et. al. 2009).
Furthermore heavy metals can replace essential metals in pigments or
enzymes disrupting their function (Malayeri et al., 2008).
3.

Molecular Mechanism of Metal Ion Toxicity

Ochaia (1987) pointed out that the molecular mechanisms of meals ion
toxicity can be divided into following five general groups (i) Displacing
essential metals ions from bio-molecules and other biologically functional
units, (ii) Inhibiting the synthesis of essential functional groups of biomolecules including enzymes and polynucleotides, (iii) Modifying the
conformation of biomolecules likes enzymes and polynucleotide, (iv)
Disrupting the integrity of biomolecules, (v) Modifying other biologically
active compounds.
4.

Basic Principles of Phytoremediation

(i)

Uptake and transport of phytoremediation technology is based

on the interaction among plant and metals. Phytoremediation
technologies can be applied only if the contaminants are in
contact with roots or uptaken by roots. Due to this plasma
membrane transporters are a subject of research for
phytoremediation technology implementation.
Heavy metal uptaken involves the same kind of transporters
which otherwise provide macro-and micronutients entrance.
Perfus-Barboech

et.

al.

(2002)

have

domostrated

the

involvement of Ca channels in Cd uptake in Arabidopsis

thalliana. The possibility of Cd mimiking Ca in plant cells can
also justify its toxicity with perturbration of metabolism and

homeostasis of vital element. Studies performed with plant cell

protoplasts have tried to ascertain if differences in transport in
sink tissues could explain the different behaviour of
hyperaccumulator plants (Cosio et al. 2004). The results
obtained with Arabidopsis halleri and Thlaspi caerulescence
show that plasma membrane of leaf cells do not account for
differences in transport. Therefore it has been hypothesized that
other mechanism may be active to direct the metals to their
subcellular compartments, where they stored; vacuoles and
lignocellulosic material such as cell wall may be among them.
Arguello et al. (2007) noticed that metal cation tranporter plays
key roles in the metabolic stress. P (1B)- ATPase are the only
subgroup of P-ATPase that contribute to heavy metal
homeostasis presented in most organisms. P (1B)-ATPase not
only mediate metal ion mobilization and uptake in roots, but
also contribute to the metals transport, storage and tolerance in
shoots especially in heavy metal hyperaccumulators.
(4.2) Accumulation and Sequestration Precism
In hyperaccumulator plants metal accumulation is completed in
two steps, firstly, plants accumulate metals in their roots and secondly
they transport these metals to arrial organs by protein transporters
(Thangavel & Subburaam, 2004; & Sinhal et al; 2010). While in nonaccumulator plants after the uptake on metal ions, they are
sequestered in vacuoles by binding with appropriate ligands like
organic acids, proteins and preptides and presence of enzymes that
can function at high level of metallic ions. Hyperaccumulator plants
usually have a shoot/root metal concentration ratio of >1, whereas
non-hyperaccumulator plants have the ration of <1 ie. non
5

hyperaccumulator plants have higher metal concentration s in roots

than in shoots (Shen et. al. 2011; Badr et al. 2012).
The standard for hyperaccumulators has not been defined
scientifically (Nazir et al, 2011). However individual authors or
research group have defined hyperaccumulator. Reeves (1992)
attempted to illustrate Ni hyperaccumulation with greater precision as
a Ni hyperaccumulator is a plant in which a Ni concentration of at
least 1000 mg kg-1 dr. wt. has been recorded in the above ground
tissue in at least one specimen growing in its natural habitat. For
establishing hyperaccumulator status, above ground tissue should be
regarded as plant foliage only. The phrase growing in its natural
habitat explains that hyperaccumulators must achieve metal
hyperaccumulation while remaining healthy enough to maintain a
self-sustaining population (Van der Ent et at. 2013). Ali et al (2013)
noticed that Baker and Brooks (1989) criteria for hyperaccumulation
is most cited criteria (with 1376 citation so far) which states that
hyperaccumulators are plant species, which accumulate greater than
100 mg kg-1 dry weight Cd, or greater than 1000 mg kg-1 dry weight
Ni Cu and Pb or greater than 10000 mg kg -1 dry weight Zn & Mn in
their shoots when grown on metal rich soils. Van der Ent et al. (2013)
admit that criteria commonly used for hyperaccumulation of some
metals are unnecessarily conservative and propose that criteria for
hyperaccumulation of such metals be lowered. They recommend the
following concentration criteria for different metals and metalloids in
dried foliage with plants growing in their natural habitats; 100 mg kg 1
for Cd, Se & Ti; 300 mg kg-1 for Co, Cu and Cr; 1000 mg kg-1 for Ni,
Pb and As; 3000 mg kg-1 for Zn; 10,000 mg Kg-1 for Mn. Van der Ent
et al. (2013) also point out that hyperaccumulator have to be recorded

from the natural habitat. They do not regard extreme accumulation

achieved through hydroponics or metal-amended spiked soils and
arficially acidified soils as hyperaccumulation. The do not consider
such experiments alone as capable of defining a species as a
hyperaccumulator. They argue that natural population must be
studied. An interesting break through that has emerged from
comparative

physiological

and

molecular

analyses

of

hyperaccumulators and related non-hyperaccumulators is that most

key steps of hyperaccumulation rely on different regulation and
expression of genes found in both kinds of plants. In particulars, a
determinant role in driving the uptake, translocation to leaves and
finally sequestration in vacuoles or cell wall of great amount of heavy
motals is played in hyperaccumulator by constitutive over expression
of genes encoding transmembrane transporters, such as members of
ZIP, HMA, MATE YSL and MTP families (Rascio 2011). Among the
hypothesis proposed to explain the function of hyperaccumulation,
most evidence has supported the elemental defense hypothesis
which states that plants hyperaccumulate heavy metals as a defense
mechanism against natural enemies such as hervivores. According to
the more recent hypothesis of joint effect heavy metal can operate
in concert with organic defensive compounds leading to enhanced
plant defense overall (Boyd, 2012).
(4.3) Biochemical mechanism of Tolerance
Plants have developed potential mechanisms at the cellular
level that might be involved in the detoxification and thus imparting
tolerance to heavy metal stress (Sunitha et al. 2014). Tolerant plant
species have evolved some mechanism to detoxify the heavy metal
stress, like immobilization of heavy metals, plasmas membrane
7

exclusion, restriction of uptake and transport of metals, synthesis of

specific heavy metal transports, chelation and sequestration of heavy
metal by ligands such as phytochelatins and metallothionoins,
induction of mechanism contrasting the effects of ROS (Reactive
Oxygen Species), induction of stress proteins, biosynthesis of proline,
polyamines and signaling molecules such as salicylic acid and nitric
oxide (Sharma and Dietz, 2009; Clemens, 2006). Genomic effects
have been promoted by the EC (European Commission) in research
project studying the properties of hyperaccumulator and results are
showing how different genes are induced by metals in those plants
and their congeners (Van De Mortal et al. 2004). Genetic mapping is
the method of choice in case of quantitative traits and some authors
are

building

maps

hyperaccumulation

of

and

quantitative
tolerance

in

trait

Loci

model

(QTLs)

plants

and

for
in

hyperaccumulators. Due to phylogenetic relationships with known

hyperaccumulators in the family Brassicaceae, Arabidopsis thaliana
is the best model plant available due to complete knowledge of the
genomic sequence and to the genetic knowledge. The group led by
Martin Broadley mapped CaTL involved in Cs accumulation of A.
thaliana (Payne et al. 2004). Several accession were analysed for Cs
accumulation, leading to the description of a 2-fold variation in Csaccumulation. Crosses among contrasting phenotypes and analysis in
segregating progenies led to mapping of putative QTLs on several
chromosomes; the existence of two QTLs on chromosomes I & V was
confirmed from the analysis of segregating populations from
indepnedent crosses. Mapping of candidate genes in these regions
will lead to new hypothesis about the structure and function of there
QTLs.

(4.4) Genetic Basis of Tolerance

According to Marmiroli et al. (2006) classical genetic studies
have been exploited in order to address the issue of genetic bases of
tolerance and/or accumulation in addition to genomic and proteomic
approaches towards gene identification. For this purpose, model
plants of choice have traditionally been Arabidopsis halleri and
Thalaspi caerulescens, two hyperaccumulators that can be crossed
with non-hyperaccumulator ecotypes or congeners for studies of traits
segregation. Genecological observations have demonstrated that
tolerance to one heavy metal is a trait which is independent from
accumulation of the same metal and also that tolerance is controlled
by few major genes. The same approach has been pursued in Thlaspi
with ecotypes differing in accumulation capacity (Zhao et al. 2003).
Segregation results suggest that 2 genes at least are responsible for Zn
accumulation, whereas for Cd accumulation more than one could be
involved. In Thlaspi Cd tolerance and accumulation segregates as
independent character.
5.

Phytoremediation Technologies
Phytoremediation exploits plants innate biological mechanism for

human benefits. Seven subsets of this technology as applicable to toxic

metal remediation from soil and water are: (i) Phytoextraction (ii)
Phytofilteration,

(iii)

Phytosabilization,

(iv)

Phytovolatilization

(v)

Phytodegradation (vi) Rhizodegradation, (vii) Phytodesalination.

(5.1) Phytoextration:
It is best approach to remove the contaminant especially heavy
metals, from soil or water by plant roots and their translocations in
above ground parts (Sinhal et al. 2010, Rafati et. al. 2011, Ali et. al.
9

2013). In metal polluted soil hyperaccumulating plants are

seeded/transplanted and are cultivated under established agriculture
methodologies.

When

maximum

plant

growth

and

metal

accumulation are achieved, plants from above ground levels are

harvested that results permanent removel of metals from the site.
(Ismail, 2012). The efficiency of phytoextraction depends on maney
factors like soil properties, speciation of heavy metals, bioavailablity
of heavy metals and plant species concerned. The plants which have
to

be

used

in

phytoextraction

should

possess

following

characteristics.
(a) Easy cultivation and harvest.
(b) Enhanced growth rate.
(c) Profusely branched root and shoot system.
(d) Widely distributed.
(e) Hyperaccumulating nature.
(f) Translocation of accumulated heavy metals from roots to shoots.
(g) Tolerant to heavy metals.
(h) Production of high biomass.
(i) Resistance to pathogens & pests.
(5.2) Phytofiltration:
In Phytofiltration plant roots (rhizofiltration) or seedlings (blasto
filtration) are grown in aerated water from where they participate and
concentrate toxic metals from contaminated effluents (Raskin et. al.
1997). In other words phytofiltration is the removed pollutants from
contaminated surface waters of or waste waters by plants
(Mukhopadhyay and Maiti, 2010). The techniques involve growing

10

plants hydroponically and transplanting into metal polluted water

from where plants absorb and concentrate the metal in their roots &
shoots. (Flathman & Lanza, 1998). The plants which have to be
utilized in rhizofiltration must posses fast growing roots with
capability for removing toxic metals from solution over extended
period of time. After saturation with metal contamination which form
precipitation over root surface, whole plants or roots are harvested for
disposal. This precipitation is caused by the root exudates and
changes in rhizospheres (Flatman & Lanza, 1998). The mechanisms
of phyto filtration are not necessarily similar for different metals.
Precipitation and exchangeable sorption are involved in case of Pb
(Ismail, 2012). The advantages of rhizofiltration include it ability tube
used as insitu or ex-situ applications and species other than
hyperaccumulators can also be used (Ghost & Singh, 2005).

(5.3) Phytostabilization:
It is also termed as phytorestoration or phytoimmobilization. It
is a technique used for stabilization of contaminants in contaminated
environment (Singh, 2012). In this remedial technique, plant
stabilizes wastes and prevent exposure pathway though wind & water
erosion, enables hydraulic control that restricts the vertical migration
of pollutants into ground

water, and immobilizes the pollutants

physically and chemically by root sorption and chemical fixation

with different soil amendments. Metals of different valances vary in
toxicity. By excreting special redox enzymes, plants skillfully convert
hazardous metals to a relatively less toxic state and decrease possible
metal stress and damage (Ali et. al, 2013). The reduction of Cr (VI) to
Cr (III) is widely studied, the latter being both less mobile and less
11

toxic (Wu et al, 2010).

The research of smith and Bradshaw (1992)

led to the developments of two cultivars of Agrosists tenuis, Sibth,

and one of Festuca rubra L. which are now commercially available
for the phytostabilization of Pb, Zn, & Cu contaminated soil. The
combination of tree and grasses work best for phyto stabilization
(Hooda, 2007). Fast transpiring tress such as Poplar maintain an
upword flow to prevent downword leaching, while grasses prevent
wind erosion and lateral runoff with dense root system. Further,
grasses do not accumulate as much metals in their shoots as dicot
species, minimizing exposure of wildlife to toxic elements (Pilon,
Smits, 2005). However phytostabilization is a temporary solution of
the problem but not permanent because heavy metals remain in the
soil. Only their environment is limited. Actually it is a management
strategy for stabilizing potentially toxic contaminants (Vangronsveld
et al., 2009).
(5.4) Phytovolatilization:
Phytovolatilization is the uptake of pollutants from soil by
plats, their conversion to volatile form and subsequent release into
atmosphere. It works well for organic pollutants but can be used for a
few inorganic that can exist in volatile form ie Se, Hg and As. The
mentioned process is controversial of all techniques due to its dubious
nature that whether release of these volatilized elements in
atmosphere is safe. The disadvantage is the volatilized element could
be recycled by precipitation and then deposit back into ecosystem
(Henry, 2000). It is fact that it does not remove the pollutant
completely, only it transfer from one segment (Soil) to on other
(atmosphere) (Padmavathiamma and Li 2007). Members of Brassica
genus and some microorganisms are particularly good volatilizers of
12

Se (Terry et al., 1992). Among the aquatic species, rice, rabbit foot
grass, Azolla and pickle weed are the best Se volatilizers (Lin
etal.2000 Zayad ital, 2000). Volatilization of Se involves assimilation
of inorganic Se into the organic selenoaminoacids selenocysteine
(SeCys) and selenomethionine (SeMet). The latter can be methylated
to form dimethylselenide (DMSe), which is volatile (Terry et al.
2000). In Hg, contaminated soil and sediments, microbial activity
converts the highly toxic Hg (II) into organomercurials and, under
optimum conditions, elemental Hg (which is far less toxic) enters the
biogeochemical cycle upon volatilization (Bizily et al. 2000). After
genetic modification of Arabidopsis thaliana L. and Nicotiana
tobaccum L. with bacterial organomercurial lyase (Mer B)

and

mercuric reductase (Mer A) genes (Rugh et al,1996, Heaton et al.

1998), plants have developed abilities to absorb elemental Hg (II)
and methyl mercury (Mer Hg) to atmosphere.
Although it is a passive process, it may be maximized by using
plant species with high transpiration rate, by over expression of
enzymes such as Cystathionine _V_ synthases that mediates S/Se
volatilization (Van Huysen et al, 2003) and by transferring gene for
Se volatilization from hyperaccumulatas to nonaccumulators (Le Duc
et al. 2004)
(5.5) Phytodegradation:
Phytodegradation is the breakdown of organics, taken up by the
plant to simpler molecules that are incorporated into plant tissues.
Phytodegradation is not dependent on rhizospheric microorganisms
(Vishnoi and Srivastava, 2008) Plants contain enzymes that can
breakdown and convert ammunition wastes, chlorinated solvent such

13

as trichloroethylene and other herbicides. The enzymes are usually

dehalogenases, oxygenases and reductase (Black, H, 1995).
According to Zozouli et al. (2014) many organic hazardous pollutants
such as bisphenol A (BPA) which are toxic and not easily
biodegradable can concerns for environmental pollution words wide.
They studied that whether Azolla filiculoides is able to remove BPA
from aquous solution. They noticed that Azolla filiculoides have 6090% BPA removal efficiency. The removal efficiency was more then
90% when BPA concentration was 5 ppm and amount of biomass 0.9
gr. They concluded that Azolla have good ability to remove the
organic compound from aquous

solution and removal efficiently

depended on the reaction time, initial BPA concentration, fern water

weight pH and temperature. The conventional method such as AOP
(Advance oxidation process) adsoption etc or high cost and consume
high energy and because all countries are faced with energy shortage
problem today; thus natural systems, for instance use of Azolla can be
good alternative to convontial system to remove of this compounds
from waste water. Recently, scientists have shown their interest in
studying phyto degradation of various organic pollutants including
synthetic herbicides & in secticides (Ali et al. 2013)
(5.6) Rhizodegradation:
Rhizodegradaion is the brakedown of organic pollutants in the
soil through soil microorganism of the rhizosphere (Mukhopadhyay
Maiti, 2010). Rhizosphere extends about 1mm around the root and is
under the influence of the plant (Pilon- Smits, 2005). The main reason
for the increased degradation of pollutants in the rhizosphere is likely
the increase in number and metabolic activities of the microbes.
Plants can stimulates microbial activity about 10-100 times higher in

14

the rhizosphere by the secretion of root exudates containing amino

acids, carbohydrates, flavonoides and amino acids According
Spaczynski (2012) soil microorganisms living in the rhizosphere play
invaluable role in the degradation of harmful organic compounds;
they are often much more involved in mineralization of xenoboiotics
than plants.
Through rhizodegradation many toxic compounds such as
pesticides,

polychlorinated

biphenyls,

polycyclic

aromatic

hydrocarbons petrolium compounds etc can easily be remove from

soil. In addition to secreting organic substrates for facilitating the
growth and activities of rhizosphere microorganicsms, plants also
provide

certain

enzymes

capable

of

degradation

organic

contaminants in soil (Kuiper et al.2004, Yadav et al .2010).

(5.7) Phytodesalination:
It is a recently reported and emerging technique to reclaim salt
affected soil (Zorrig et al. 2012). Phytodesalination refers to the use
of halophytic plants for removal of salts from salt effected soils in
order to enable them for supporting normal plants growth (Manousaki
and Kalogerakis, 2011; Sakai et al; 2012). It was postulated that salt
affected soil cover about 6% (more than 800 million ha) of the world
lands, which is mainly due to natural causes (Salt accumulation over
long period of time in arid and semi arid regions) or to humaninduced causes that affected in 2008 about 2% (32 million ha) of the
dry land formed areas and 20% (45 million ha) of the irrigated lands
in world (Zorrig et al; 2012). To overcome this problem, several
authors (Ravindran et al. 2007), Rabhit et al. 2009; Rabhi et al.
2010a; Rabhi et al. 2010b)

have been encouraging this biological

approach proving the efficiency of Na + -hyper accumulating plants to

15

desalinize the soil on which they are cultivated, especially in arid and
semiarid regions, where low precipitations expectations and
inappropriate irrigation system are unable to leach salts from the
rhizosphere. Soil phytodesalition is based on the capacity of some
halophytes to accumulate enormous sodium quantities in their shoots
(Rabhi, 2010b). Halophytic plants have been suggested to be
naturally better adapted to

cape with heavy metals compared to

glycophytic plants ( Manousaki and Kalogerakis 2011). According to

an estimation two halophytes, Suaeda maritima and Sesuvium
portulacastrum could remove 504 and 474 kg of sodium chlorode
(Nacl) respectivety from 1 hactare of saline soil in a period of 4
months. Therefore, S. maritima and S. portulacastrum could be
successfully used to accumulate Nacl from highly saline soils and
enable them for crop production after a few repeated cultivation and
hervest (Ravindran et al.2007).
6.

Induced Phytoextration or Chelant assisted PhytoextractionThe term chelate denotes a complex between metal and a chelating

agent and not the chelating agent itself. (Nowack and Van Briesen, 2005). A
shorter word for chelating agent is chelant or chelator. It is therefore
suggested for using the term chelant- enhanced phytoextraction. The
bioavailability of heavy metals in soil is a critical factor affecting the
efficiency of phytoextraction of target heavy metals. Low bioavailability of
heavy metal is a major limiting factor for phytoextraction. In induced or
chelate- assisted phytoextraction, different chelating agents life EDTA,
HEDTA, NTA, citric acid, elemental sulfure and ammonium sulfate are
added to soil to increase the bioavailability of heavy metals in soil for
upkate by plants (Lone et al. 2008, Sinhal et al. 2010, Sun et al, 2011b).
Sinhal et al. (2010) studied the effect of EDTA and citric acid on phyto
16

extraction potential of marigold to Zn, Cu,Pb, and Cd, and noticed that
EDTA

and

citric

acid

both

efficiently

enhanced

many

fold

phytoextractability of marigold to these metals. Wenzel et al. (2003)

hypothesized that free protonated EDTA enters the roots, subsequently
forming metal complexes that enhance metal transport to shoots. Lai et al
(2004) also suggested that EDTA forms metal-EDTA complex in soil which
becomes readily available for uptake and translocation to aerial parts of the
plants. Turgut et al. (2005) investigated the effect of citric acid on heavy
metal (Cd, Cr and Ni) uptake and translocation in Halianthus annuus us and
found that citric acid significantly increased the metal availability and
enhanced the metal accumulation many folds in shoots of plant. For the
chelant tested, the order of effectiveness in increasing Pb adsorption from
the soil was EDTA (ethylene-diamine tetra acetic acid) > HEDTA (Hydroxy
ethylene diaminetriacetic acid) > EDDHA (Ethylenediamine di (ohydroxyphenylacetic acid) (Hung et al. 1997). Bioavailability of the heavy
metals can also be increased by lowering soil pH since metal salts are
soluble in acidic media rather than in alkaline media. (Ali et al, 2013).
However, these chemical treatment can cause secondary pollution
problems. For example, synthetic chelant EDTA is non-biodegradable and
can leach into ground water supplies making an additional environmental
hazards. Furthermore, synthetic chelating agents can also be toxic to plants
at elevated levels (Sinhal et al. 2010). Thus proper care should be taken
when practicing induced phytoextraction (Zhao et al. 2011; Song et el.
2012).

17

7.

Quantification of Phytoextraction Efficiency

The phytoextractability or phytoexctraction efficiency of plants can

be calculated by bioconcentration factor and translocation factor.

Bioconcentration factor denotes the efficiency of a plant species to
accumulate heavy metals from surrounding environment. It can be calculate
as follows (Zhuang et al. 2007)
Bioconcentration Factor (BCF)

= Concentration of Metal in Harvested plant tissue

Concentration of some metal in soil

Translocation Factor denotes the efficiency of a plant species to

translocate accumulated metals from root to shoot. It can be calculate as
follows (Padmavathiamma & Li, 2007).
Translocation Factor (CTF)

= Concentration of Metal in shoots

Concentration of Metal in roots

Shahid et. al. (2014) studied the long term field metal extraction by
Pelargonium and its phytoextraction efficiency and noticed that lead (Pb)
was the element most efficiently transferred from soil to the plant, following
by Cd, Zn. Cu and As with BCF (Shoot) values of 0.22, 0.07, 0.05, 0.04,
0.01 and 0.05 respectively.
These low BCF (Shoot) values were due to very high concentrations
in the bulk soil, indeed. According to Liu et al. (2010) and FrancoHernandez et al. (2010), BCF (Shoot) values of heavy metals decrease with
increasing metal concentrations in the soil. Shahid et al. (2014) noticed that
most of the lead absorbed by roots is translocated to arial parts
(Translocation Factor = 1.6). Enhanced metal uptake from roots and
translocation to the shoots in hyperaccumulators in generated by specific
18

carrier protein members. Maestri et al. (2010) reviewed several studies that
reported the presence of metal transporters such as ZIP (ZRT/IRT-like
protein), CDF (cation diffusion facilitator) and HMA (Heavy metal ATPare)
among plant cells. According to Yoon et al. (2006), only plant species with
both BCF and TF greater than 1 have the potential to be used for
phytoextraction. Hyperaccumulators have BCF greater that 1 sometimes
reaching 50-100 (Cluis, 2004). BCF might have use for comparisions in
case of growing plants in homogenized soil or in hydroponic culture but has
little advantage over simple comparison of foliar metal concentration
(Vander Ent et al. 2013)
8.

Genetic Engineering to improve phytoremediation

To breed plants having superior phytoremediation potential with high

biomass production can be an alternative to improve phytoremediation.

Considerable progress has been made recently in identifying plant genes
encoding metal ion transporters and their homologous in hyperaccumulator
plants. Therefore, it is hoped that genetic engineering may offer a powerful
new means by which to improve capacity of plants to remediate
environmental pollutants (Yang et al. 2005a, Mello-Farias & Chaves, 2008).
Brassica juncea was genetically engineered to investigate rate-limiting
factors for glutathione and phytochelatin production. To achieve this, in
Escherichia coli gshl gene was introduced. The Y-ECS transgonic seedlings
showed increasged tolerance to Cadmium and had higher concentrations of
phytochelatins, y-GluCys, glutathione, and total non protein thiols
compared to wild type seedling (Fulekar et al. 2008). Some genes have been
isolated and introduced into plants with increased heavy metal (Cd)
resistance and uptake, like At Nramps (Thomine et al. 2000), At Pcrs (Song
et al. 2004). CAD1 (Ha et al., 1999) from Arabidopsis, thaliana, library
enriched in Cd induced cDNAs from Datura innoxia (Louie et al, 2003) and

19

PCS CDNA clone (Heiss et al. 2003) from Brassica juncea. There are some
examples of transgenic plants for metal tolarance/phytoremediation, as
tobacco with accumulation of Cd, Ca, and Mn transformed with gene CAX2 (vacuolar transporters) from Arabidopsis thaliana (Hirschi et. al. 2000);
A. thaliana tolerant to Al, Cu, and Na with gene Glutathione-S-transfer ase
from tobacco (Ezaki et al. 2000) and rice (Goto et al. 1998; 1999) with
increased iron accumulation with gene Ferrethin from soybean. New
metabolic pathway can be introduced into plants for hyperaccumulation or
phytovolatilization in case of Mer A and Mer B genes which were
introduced into Arabidopsis and resulted into several fold increased
tolerance to Hg and volatilization of elemental mercury (Dhankher et al.
2002; Eapen & D Souza, 2005) by transgenic Arabidopsis.
9.

Utilization of phytoremediation by products

Phytoextraction involves repeated cropping of plants in contaminated

soil, until the metal concentration drops to acceptable levels. Theoretically,

metal removal can be accounted by determining metal concentration in
plant, multiplied by the biomass products, and comparing this with the
reduction in soil metal concentrations. Although this sounds simple, many
factors make it challenging in the disposal of contaminated plant material
(Jadia and Fulekar, 2011). After each cropping, the plant is removed from
the site; this leads to accumulation of huge quantity of hazardous biomass.
This hazardous biomass should be stored or disposed appropriately so that it
does not pose any risk to the environment. Composting and compaction has
been proposed as post harvest biomass treatment by some authors (Garbisu
& Alkorta 2001). Experiment carried out by Hetland et al. (2001), showed
that composting can significantly reduce the volume of harvested biomass,
however metal contaminated plant biomass would still require treatment
prior to disposal. Compaction of harvested plant material was proposed by
20

Blaylock and Huang (2000) for processing metal rich phytoextraction

residue.
Combustion and gasification are the most important sub-routes for
organized generation of electrical and thermal energy. Recovery of this
energy from biomass by burning or gasification could help make
phytoextraction more cost effective. Thermochemical energy conversion
best suits the phytoextraction biomass residue because it cannot be utilized
in any other way as fodder and fertilizers. Combustion is a crude method of
burning the biomass, but it should be under controlled conditions whereby
volume is reduced to 2-5% and ash can be disposed proerly. Gasification is
the process through which biomass material can be subject to series of
chemical changes to yield clean and combustive gas at high thermal
efficiencies. The process of gasification of biomass in a gasifier is a
complex

phenomenon; it involves drying, heating thermal decomposition

(pyrolysis) and gasification and combustion chemical reactions, which

occurs simultaneorly (Iyer et al; 2002). Bridgewater et al. (1999). reported
that pyrolysis is a novel method of municipal waste treatment that might
also be used for contaminated plant material. Pyrolysis decomposes material
under aerobic conditions; there is no emission to the air. The final products
are pyroltic fluid oil and coke; heavy metals will remain in the coke which
could be used in smelter. Helson et al. (1997) conducted low temperature
pyrolysis experiment with chromium copper and arsenate treated wood and
it was concluded that most of the metal was retained in pyrolysis residue.

21

10.

Limitations of Phytoremediation
Although phytoextraction is a cost effective promising approach for

remediation of heavy metal contaminated sites. It is most effective if soil

contamination is limited to within 3 feet of the surface and if ground water
is within 10 feet of the surface (Cunninghum et al. 1997). It is applicable to
sites with low to moderate soil contamination over large areas, and to sites
with large volumes of ground-water with low levels of contamination that
have to be cleaned to low (strict) standers. Furthermore, there are some
more limitations to phytoextraction (Maiti, 2010, Sinhal et al. 2010. Naess
et al. 2011, Ramamurthy and Memarian, 2012)
- High concentration of hazardous materials can be toxic to plants.
- It may be seasonal, depending on location.
- Required long time to clean-up the contaminated sites.
- It can transfer contamination across media, eg. from soil to air.
- It is not effective for strongly sorbed (Example. PCBs) and weakly
sorbed contaminants.
- The toxicity and bioavailability of biodegradation products is not
always known.
- Products may be mobilized into ground water or bio-accumulated
in animals.
- It is still in the demonstration stage.
11.

Future trends in Phytoremediation

One of the key aspects to the acceptance of phytoremediation pertains

to the measurement of its performance, ultimate utilization of by-products

and its overall economic viability. Currently most research is limited to
22

laboratory and greenhouse scale studies and only a few studies have been
conducted to test the efficiency of phytoextraction in actual field. There are
many factors, that may affect phytoremediation in field, viz. variation in
nutrients, temperature, soil pH, precipitation, moisture and uneven
distribution of contaminants (Vangronsveld et al., 2009). Genetic evaluation
of hyperaccumulators growing in metal contaminated soil and associated
microbes would provide the researchers with a gene pool to be used in
genetic manipulation of other non-accumulators and production of
transgenics (Hooda, 2007). Phytoremediation efficiency of different plants
for specific target heavy metals has to be tested in field conditions in order
to realize the feasibility of this technology for commercialization. (Ali et al.
2013). Commercially to enhance public acceptance phytoremediation can be
integrated with landscape architecture such as remediation of partially
contaminated urban sites may be combined with an attractive design so that
the area may be used as a park or some other recreational place by the
public after remediation process. There is need to optimizing the plants for
phytoremediation and to gain new knowledge about the fate and transport of
metals/metalloid in plants and discovery of new hyperaccumulators plants
and their proper management.
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40

Table: List of Hyperaccumulator Plants

Sl.No.

Plant Species

Metal

Metal
Accumulation
(mg kg-1)

Reference

1.

Alyssum murale

Ni

4730-20100

Bani et al (2010)

2.

Arabis paniculata

Cd

1127

Zeng et al (2009)

3.

Astragalus racemosus

Se

14,900

Beath et al. (1937)

4.

Brassica juncea

Ni

3916

Saraswat and Rai (2009)

5.

Berkheya coddii

Ni

18000

Mesja sz-Przybylowicz et. al (2004)

6.

Corrigiola telephiifolia

As

2110

Garcia Salagado et al (2012)

7.

Eichornia crassipes

Cr

6000

Lytle et al. (1998)

8.

Eleocharis acicularis

Cu

20200

Sakakibera et al (2011)

9.

Euphorbia cheiradenia

Pb

1138

Chehregani and Malayeri (2007)

10.

Ipomea alpina

Cu

12,300

11.

Isatis pinnatiloba

Ni

1441

Altinozlu et al (2012)

12.

Iberis intermedia

Ti

3070

Leblana et al. (1999)

13.

Potentilla griffithii

Zn

19,600

14.

Phragmites australis

Cr

4825

Calheiros et al. (2008)

15.

Pteris vittata

As

8331

Kalve et al. (2011)

16.

Rorippa globosa

Cd

>100

Wei et al. (2008)

17.

Sesbania drummondi

Cd

1687

Israr et el (2006)

18.

Sedum alfredii

Zn

18,799

Jin and Liu (2009)

19.

Sedum alfredii

Cd

2183

Jin and Liu (2009)

20.

Sorgham sudanense

Cu

5330

Wei et al (2008)

21.

Thlaspi calrulescons

Zn

10410

Banasova and Horak (2008)

22.

Tegetus crecta

Zn

526

Sinhal et al (2010)

23.

Tegetus crecta

Pb

393

Sinhal et al (2010)

24.

Tegetus crecta

Cd

333

Sinhal et al (2010)

Baker and Walker (1989)

Hu et al. (2009)

41

Table-2: Phytoremediation includes the following process

and mechanisms of contaminant

Sl.No.

Process

Mechanism

Contaminant

1.

Phytoextraction

Hyperaccumulation

Inorganics

2.

Phytofilleration

Sequesteration of pollutants

Organics/Inorganics

3.

Phytovolatilization Volatlization of Pollutants by

Leaves

Organics/Inorganics

4.

Phytostabilization Complexation

Inorganics

5.

Phytodegradation Degradation of organics in plant Organics

tissues

6.

Rhizodegradation Degradation of organics in

rhizosphere

Organics

7.

Phytodesalination Removal of excess salts by

halophytes

Inorganics

42

Soil
Chemistry
Plant Molecular
Biology

Environmental
Engineering

Soil
Microbiology

Phytoremediation

Ecology

Environmental
Toxicology

Plant
Genetics
Environmental
Biotechnology

Schematic representation of interdisciplinary nature of Phytoremediation research

Fig. 1

43