SLIDE 2-Plants interact with the environment through signalling compounds that code for

their unique language. Chemical signals are widely distributed in plant and animal kingdom
and is secreted from one organism and affect other organisms in the neighbourhood and play
very important roles in sustainable ecosystems, such as dominance, succession and climax of
plant community, biodiversity, crop productivity and pollution abatement and restoration of
environment
SLIDE 3The use of plants to reduce contaminants and restore the soil resource is a cost-
effective method of reducing the risk to human and ecosystem health posed by contaminated
soil sites. And this phenomenon is termed phytoremediation.
Phyto means plant and remedium means restoring balance, defined as “the efficient
use of plants to remove, detoxify or immobilize environmental contaminants in a
growth matrix (soil, water or sediments) through the natural biological, chemical or
physical activities and related processes of the plants”.
Plants remove contaminants from the environment in various ways-
Phytoextraction – Also called phytoaccumulation, in which hyperaccumulator plants absorb
metals from soil through the root system
Phytostabilization – Also refers to as in-place inactivation. Plants immobilise contaminants in
the soil and groundwater through absorption and accumulation by roots, adsorption onto
roots,
Phytodegradation - It is also known as phytotransformation. It is the breakdown of
contaminants taken up by plants through metabolic processes within the plant or the
breakdown of contaminants external to the plant through the effect of compounds produced
by the plants.
Phytovolatilization – Uptake and transpiration of a contaminant by a plant, with the release of
contaminant or a modified form to the atmosphere.
Rhizofiltration – filtering water through a mass of roots to remove toxic substances. The
pollutants remain absorbed in or adsorbed onto the roots.
Here we observe tht phytoextrct and phytostabilize ad also rhizofiltrat are mechanism by
which plants decontaminate inorganic polltants eg heavy metals while through
phytodegradation and volatilization , organic pollutants such as petroleum by products can be
removed

SLIDE 5 It is an undemanding crop plant, by undemanding we mean tht it is not grown as a

commercial crop plant, but it has attributes which can be used in the commercial purposes.

Glucosinolates (GS) are a class of plant secondary metabolites that provide defense against
herbivores and may play an important role in pollinator attraction. But along with these roles,
it is observed that Exposition of A. thaliana a member of brassicaceae to cadmium ions leads
to the increased expression of the genes for glutathione synthetase (GS). All these findings
support importance of glucosinolates in cadmium detoxification (Lee et al. 2003).

SLIDE 6 Based on the some unrelated research done mostly on genome duplications to trace
the evolution of Cleomaceae and Brassicaceae, it was reported by Erik van den Bergh 2016,
that the Many Glucosinolate biosynthesis genes have expanded through polyploidy, gene
transposition and duplication, in Cleomaceae.
Therefore from the genetic point of view we tried searching new plants belonging to this less
studied sister family of Brassicaceae, to have phytoremediating capabilities

SLIDE 7 There are 3 members of Cleomaceae presently recorded in India. C. viscosa, C,

gynandra and C. rutidosperma . Among these, the predominant species that is found
ubiquitously C. rutidosperma. It is a common Annual herb, up to 1 m tall, that grows as a
weed in disturbed and ruderal habitats, principally in areas around local sewage disposal
areas disturbed ground, roadsides, gardens, crops and abandoned lands,. It has a wide range
of medicinal properties like anti helminthic, antidiabetic, wound healing. even
antitumourogeinc properties are also reported, supporting its age old use as a medicinal plant
in Tropical Africa.

SLIDE 10 Removal of Those metals are important since they are capable of decreasing crop
production due to the risk of bioaccumulation and biomagnification in the food chain. There’s
also the risk of superficial and groundwater contamination.

SLIDE 11On basis of the ref , the threshold concentration criteria for different metals and
metalloids in hyperaccumulator tissue : 100 ug/g for Cd, 1,000 ug/g for Pb; 3,000 ug/g for
Zn. Normal Cd levels are so low (0.03-5.0 ug/g in most plant species) that those plants
capable of concentrating this element to >100 ug/g should be regarded as Cd
hyperaccumulators (Baker et al. 1994a), having significant phytoremediation potential.

SLIDE 20 When compared to the threshold values mentioned by reeves et al, C. rutidosperma
can be a competent accumulator of Cd and Zinc, but does not match the value for Pb.

SLIDE 13 The 2nd part of my 1st objective comprises of the pot culture experiment. Here is
the general lay out of my experimental design. Completely randomised design was followed
with cdcl2 as the factor. normal garden soil was taken for the experiment. 6 diiferent
treatments were implied that are as follows. T1 contains only the soil witout any external
amendments and is considered as control. T2 to T6 contains CDCL2 concentrations ranging
from 10 to 200 mg/kg. thye pot size was ……. . 3 replicates were done fr each tyreatment and
10 seedlings per set were planted. A total of 18 sets were considered.

SLIDE 14

SLIDE 15 to study the effect of cadmium on the phytophisiology, mainly 2 parameters were
considered change in biomass.

SLIDE 18 Here we can see that as compared to the control set, both fresh wt and dry wt are
not significantly different both in case of root and shoot. But an interestingly it was observed
that in case of 20ppm conc , there is a significant increase in the fresh and dry wt. this can be
explained by the fact that when a plant is in stress the plant tries to cope it up by enhancing its
recovery mechanism so much that the growth rate by the end results to be better than normal.
While when the stress still continues the plant defence sysytm normalizes and the growth
returns back to normal that is comparable to the control set. reference

Further analysis of parametrs are to be considered to clarify this reason like proline content,
etc.
But in case of the total chlorophyll content we do not see any of such trend. Here we can see
that there is no significant difference in the sample sts wrt to control set.

SLIDE 19 HERE we see that the roots have accumulated metal linearly wrt to conc , but in
case of the accumulation of cd in aerial parts of the plant, we observed that at the highest
concentration applied i.e 200ppm the plant can accumulate cd only and significantly higher
than all the other concentrations. This signifies that the plant has a typical threshold value
after which the plant tends to translocate the metal to its aerial parts but before it is reached
the plant tends to store the metal in its root itself thus immobilizing the same.

SLIDE 20 If we look at the linear relationship of the following based on the dosage conc. In
case of cd uptake by roots of c rutidosperma it is posivly and strongly correlated with the
conc of treatments while the uptake by aerial parts is not strongly correlated with the dosage ,
but even then if we consider the percentage removal of cd from the soil, we can see that it is
also positively and striongly correlated with the conc treatments. This implies that the plant
has a dose responsive manner of uptake of cd, the more is the conc of cd in the immediate
rhozosphere , the more efficient is the hyperaccumulation abilty of the plant .

SLIDE 21 Now if we look at the concentration factors indices that determine the class of the
plant as a phytoremediator species. Here for c rutidopserma we can see that BCF and BAF
both are much greater than 1 but in case of TF only at 200ppm the TF is more than 1 which
again states the fact that plant cant be called as a phytostabilizer in lower contaminated areas
but it can act as hyperaccumulator in areas which highly contaminated with cd.

Plants can increase their bioavailability by releasing a variety of root

exudates, which can change rhizosphere pH and increase heavy metal
solubility (Dalvi and Bhalerao, 2013). The bioavailable metal is sorbed at
the root surface and moves across the cellular membrane into the root cells.
The uptake of heavy metals into roots occurs mainly through two pathways,
apoplastic pathway (passive diffusion) and symplastic pathway (active
transport against electrochemical potential gradients and concentration
across the plasma membrane). The common uptake of heavy metals via
symplastic pathway is an energy-dependent process mediated by metal ion
carriers or complexing agents

Generally, there are two defense strategies adopted by plants to cope with
the toxicity of heavy metals: avoidance and tolerance. By these two
mechanisms, plants manage to maintain the cellular concentrations of
heavy metals below the toxicity threshold levels (Hall, 2002).

t works as the first line of defense at extracellular level through a range of

mechanisms such as root sorption, metal ion precipitation, and metal
exclusion (Dalvi and Bhalerao, 2013). Upon exposure to heavy metals,
plants first try to immobilize them either through root sorption

Once the heavy metal ions get entry into the cytosol, tolerance strategy is
adopted by the plants to cope with the toxicity of accumulated metal ions. It
is the second line of defense at intracellular level through various
mechanisms such as inactivation, chelation, and compartmentalization of
heavy metal ions (Dalvi and Bhalerao, 2013).

This is mainly achieved through chelation by complexation of heavy metal

ions with ligands. Through chelation, the concentrations of free metal ions
are reduced to relatively low levels. There are many organic and inorganic
ligands in the cytoplasm that mediate heavy metal chelation. The organic
compounds involved in heavy metal ion chelation include organic acids,
amino acids, phytochelatins (PCs), metallothioneins (MTs), and cell wall
proteins/pectins/polyphenols (Hall, 2002; Sharma and Dietz, 2006; Gupta
et al., 2013b). 

o improve phytostabilization efficiency, organic or inorganic amendments

can be added to the contaminated soil. These soil amendments can alter
metal speciation, reduce heavy metal solubility and bioavailability by
changing pH value and redox status of the soil (Alvarenga et al.,
2009; Epelde et al., 2009; Burges et al., 2018). Moreover, the application of
amendments can increase the organic matter content and essential
nutrients of the soil and improve physicochemical and biological
properties, which can benefit plant colonization and improve water-holding
capacity.
Interestingly, microorganisms living in the rhizosphere, such as bacteria
and mycorrhiza, can assist phytostabilization. These microorganisms can
improve efficiency of heavy metal immobilization through adsorbing metals
onto their cell walls, producing chelators and promoting precipitation
processes (Göhre and Paszkowski, 2006; Mastretta et al., 2009; Ma et al.,
2011). They can also increase plant root surface and depth to facilitate
phytostabilization and even serve as a filtration barrier against heavy metal
ion translocation from roots to shoots (Göhre and Paszkowski, 2006).

The plant species for phytoextraction should possess the following

characteristics: (i) high tolerance to the toxic effects of heavy metals, (ii)
high extraction ability with accumulation of high levels of heavy metals in
aboveground parts, (iii) fast growing with high biomass production, (iv)
abundant shoots and extensive root system, (v) good adaptation to
prevailing environment, strong ability to grow in poor soils, easy cultivation
and harvest, (vi) highly resistant to pathogens and pests, be repulsive to
herbivores to avoid heavy metals entering into the food chain (Seth,
2012; Ali et al., 2013). Generally, hyperaccumulators are plant species
capable of accumulating very high levels of heavy metals in their
aboveground parts without phytotoxicity symptoms (Rascio and Navari-
Izzo, 2011; van der Ent et al., 2013).  The naturally occurring heavy metal
hyperaccumulator can accumulate metals at levels 100-fold greater than
common non-hyperaccumulating species under the same conditions
(Rascio and Navari-Izzo, 2011). Strictly, the definition of hyperaccumulator
should meet the following criteria: (1) the shoot-to-root ratio of heavy metal
concentration is greater than 1, which is a sign of efficient ability to
transport metals from roots to shoots (McGrath and Zhao, 2003; Marques
et al., 2009); (2) the shoot-to-soil ratio of heavy metal concentration is
greater than 1, indicating a higher capability to take up heavy metals from
soil (McGrath and Zhao, 2003); and (3) the concentration of the metal in
the shoot is higher than 10 mg/kg for Hg, 100 mg/kg for Cd and Se, 1,000
mg/kg for Co, Cu, Cr, Ni, and Pb, and 10,000 mg/kg for Zn and Mn (Baker
and Brooks, 1989).

Searching for effective hyperaccumulators is a key and the most

straightforward strategy for successful phytoremediation of heavy metals.
Currently, more than 450 plant species from at least 45 angiosperm
families have been identified as metal hyperaccumulators so far (Suman et
al., 2018), ranging from annual herbs to perennial shrubs and trees, such as
Brassicaceae, Fabaceae, Euphorbiaceae, Asterraceae, Lamiaceae, and
Scrophulariaceae families (Salt et al., 1998; Dushenkov, 2003). Some
species can even accumulate more than two elements, such as Sedum
alfredii, which can hyperaccumulate Zn, Pb, and Cd (He et al., 2002; Yang
et al., 2002, 2004). A list of some plants, which show high capacity of heavy
metal accumulation is given in Table 1. However, using edible crops for
phytoremediation should be avoided as heavy metals can accumulate in
edible parts of the plant and thus enter into the food chain by human or
animal consumption, raising concerns on human health. Hence, selection
of the non-edible hyperaccumulators is a key for efficient and safe
phytoremediation of heavy metals.

Use of plant-associated microorganisms (rhizosperic microorganisms) is

another approach to improve plant performance for phytoremediation. The
microbial community of the rhizosphere may directly stimulate root
proliferation and, thus, promote plant growth, increase heavy metal
tolerance and plant fitness (Gupta et al., 2013a; Fasani et al., 2018).
It has been shown that plant growth-promoting rhizobacteria (PGPR) have
large potential to improve phytoremediation efficiency. PGPR can promote
plant growth and fitness, protect plants against pathogens, increase plant
tolerance to heavy metals, improve plant nutrient uptake as well as heavy
metal uptake, and translocation (Ma et al., 2011). It has been reported
elsewhere that microorganisms in the rhizosphere significantly increase
heavy metal availability and uptake by plants (Vamerali et al.,
2010; Sheoran et al., 2011). These microorganisms can secrete enzymes and
chelate into the rhizosphere, which lead to the formation of heavy metal–
chelate complexes, thus improving heavy metal uptake and translocation
(Clemens et al., 2002).