Appl Biochem Biotechnol (2012) 167:1550–1559

DOI 10.1007/s12010-012-9657-0

The Assessment of Phytoremediation Potential of Invasive

Weed Amaranthus spinosus L.

M. Devi Chinmayee & B. Mahesh & S. Pradesh & I. Mini &

T. S. Swapna

Received: 30 October 2011 / Accepted: 19 March 2012 /

Published online: 19 April 2012
# Springer Science+Business Media, LLC 2012

Abstract Presently the environment is heavily polluted by various toxic metals, which
creates danger for all living beings. Heavy metals are toxic above certain threshold
levels. Phytoremediation is an emerging technology which is quite a novel technique
of cleaning polluted sites through the use of plants. Phytoremediation methods are
comparatively cheap and ecologically advantageous, compared to conventional and
physicochemical methods like precipitation, evaporation and chemical reduction. In
this respect, plants can be compared to solar-driven pumps capable of extracting and
concentrating certain elements from their environment. Amaranthus spinosus, an
invasive weed seen on road sides and bare land belonging to the family Amarantha-
ceae, was selected for the present study. A greenhouse experiment was conducted and
consisted of a range-finding test and definitive test for various concentrations of heavy
metals Cu, Zn, Cr, Pb and Cd. Plants were grown in soil treated with different
concentration of metals depending upon the threshold level. The bio-organics of the
plant such as soluble sugar, protein, lipid, phenol, amino acid and photosynthetic
pigments were estimated after 30 days of treatment. The bio-organics showed pro-
found variation in response to accumulation of heavy metals. Accumulation of Cu, Pb and Cd
was high in the roots followed by stem and leaves and that of Zn and Cr remained high in aerial
parts. A steady increase was noticed in the bioaccumulation of copper, zinc and cadmium on
enhancing the concentration of the corresponding metal in the soil. The bioconcentration factor
and translocation factor were above unity in most of the treatments and increased as the
concentration of treatment increased which indicated that A. spinosus is a potential agent for
heavy metal accumulation and translocation.

Keywords Phytoremediation . Amaranthus spinosus . Amaranthaceae . Heavy metals .

BCF . TF

M. D. Chinmayee : B. Mahesh : S. Pradesh : I. Mini : T. S. Swapna (*)

Department of Botany, University College Trivandrum, Thiruvananthapuram, Kerala, India
e-mail: drswapnats@gmail.com
Appl Biochem Biotechnol (2012) 167:1550–1559 1551

Introduction

Phytoremediation refers to use of plants to remove pollutants from the environment or

to render them harmless [1]. Green plants are not only the lungs of nature with an
ability of purifying impure air by photosynthesis, but some species also have the
unique ability to uptake, tolerate and even hyperaccumulate heavy metals and other
toxic substances from soils and water through roots and concentrate them in roots,
stem and leaves [2]. The heavy metal accumulation capacity of plants belonging to
the Amaranthaceae family was reported earlier. Amaranthus hybridus accumulated Pb,
Cd, Hg, Ni, Mn and Fe [3]. Amaranthus retroflexus, Gomphrena globosa and Celosia
argentea were identified as metal accumulators [4, 5]. Amaranthus tricolor showed
high cadmium-accumulating ability [6], and Alternanthera sessilis growing on sewage
sludges were found appropriate for metal accumulation [7]. The phytoremediating
capacity of weeds has emerged as their most effective and useful role which can be
elaborated and utilised in tackling the problem of metal contaminants in the biosphere.
The present study helps to assess the phytoremediation efficiency of an invasive
terrestrial weed Amaranthus spinosus L. under the stress of selected heavy metals
with special reference to the impact of heavy metals on bio-organics of the plant.
Reports on the impact of heavy metals on the bio-organics of A. spinosus remain few
and fragmentary which prompted the present investigation. Heavy metals selected for
study include two essential (copper and zinc) and three toxic (cadmium, lead and
chromium) elements.

Materials and Methods

Bioassay Pot culture method was adopted for the bioassay. Healthy plants of A.
spinosus were collected from its natural habitat. Threshold level of each metal was
estimated, and three different treatment concentrations (T1, T2, T3) of each metal salt

Sugar content in Amaranthus spinosus under

heavy metal stress
450
Concentration mg g-1

400
350
300
250
200
150 Leaf
100 Stem
50 Root
0
Control

T1
T2

T3

T1

T2

T3

T1
T2

T3

T1

T2

T3

T1
T2

T3

Zn CU Cd Pb Cr
Treatments

Fig. 1 Sugar content in A. spinosus under heavy metal stress

1552 Appl Biochem Biotechnol (2012) 167:1550–1559

Protein content in Amaranthus spinosus under

heavy metal stress
Concentration mg g-1 100
90
80
70
60
50
40
Leaf
30
20 Stem
10
Root
0
Control
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
Zn CU Cd Pb Cr
Treatments

Fig. 2 Protein content in A. spinosus under heavy metal stress

were applied to soil. Garden soil was collected and uniformly saturated with varying
concentrations of copper sulphate (60, 200 and 300 mM), zinc sulphate (200, 280 and
350 mM), lead nitrate (30, 90 and 180 mM), chromium chloride (60, 180 and
360 mM) and cadmium sulphate (20, 25 and 30 mM). The test plants were grown
in pots containing 2 kg garden soil saturated with varying concentrations of metal
solution. Untreated soil was used to raise control plants, and the biochemical param-
eters were estimated after 1 month.

Analytical Techniques Bio-organics comprise a wide variety of biologically significant

macromolecules such as pigments, primary and secondary metabolites which characterize
and regulate various biological processes. Bio-organics in plant samples were estimated

Lipid content in Amaranthus spinosus under

heavy metal stress
35
Concentration in %

30

25

20

15
Leaf
10
Stem
5
Root
0
Control

T1

T2

T3

T1

T2

T3

T1

T2

T3

T1

T2

T3

T1

T2

T3

Zn CU Cd Pb Cr
Treatments

Fig. 3 Lipid content in A. spinosus under heavy metal stress

Appl Biochem Biotechnol (2012) 167:1550–1559 1553

Phenol content in Amaranthus spinosus under

heavy metal stress
2
1.8
Concentration mg g-1

1.6
1.4
1.2
1
0.8 Leaf
0.6
Stem
0.4
Root
0.2
0
Control
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
Zn CU Cd Pb Cr
Treatments

Fig. 4 Phenol content in A. spinosus under heavy metal stress

following standard procedures. Bio-organics in the control and treated plants such as total
soluble sugar [8], total soluble protein [9], total lipids [10], total phenol [11] and photosyn-
thetic pigments [12] were analysed.

Heavy Elements Estimation of the heavy metals (copper, zinc, cadmium, lead and chromi-
um) was carried out by subjecting the sample to wet digestion using a mixture of

Aminoacid content in Amaranthus spinosus

under heavy metal stress
4.5

4
Concentration mg g-1

3.5

2.5

2
Leaf
1.5
Stem
1 Root
0.5

0
Control

T1

T2

T3

T1

T2

T3

T1

T2

T3

T1

T2

T3

T1

T2

T3

Zn CU Cd Pb Cr
Treatments

Fig. 5 Amino acid content in A. spinosus under heavy metal stress

1554 Appl Biochem Biotechnol (2012) 167:1550–1559

Chlorophyll a content in Amaranthus spinosus

under heavy metal stress
Concentration mg g-1 0.018
0.016
0.014
0.012
0.01
0.008
0.006 Leaf
0.004 Stem
0.002
0
Control
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
Zn CU Cd Pb Cr
Treatments

Fig. 6 Chlorophyll a content in A. spinosus under heavy metal stress

concentrated nitric acid and perchloric acid (4:1) for 8 h and aspiring in atomic absorption
spectrophotometer [13]. The concentration of various heavy metals was computed and
expressed as milligrams per kilogram.
All experiments were performed in triplicate, and standard deviation, bioconcentration
factor [14] and translocation factor [15] were also computed.
Bioconcentration factor ¼ Metal concentrationin the plant tissue=Metal concentration in the soil

Translocation factor ¼ Metal concentration in stem and leaf =Metal concentration in root

Chlorophyll b content in Amaranthus spinosus

under heavy metal stress
0.012
Concentration mg g-1

0.01

0.008

0.006

0.004 Leaf
Stem
0.002

0
Control
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3

Zn CU Cd Pb Cr
Treatments

Fig. 7 Chlorophyll b content in A. spinosus under heavy metal stress

Appl Biochem Biotechnol (2012) 167:1550–1559 1555

Total Chlorophyll content in Amaranthus

spinosus under heavy metal stress
0.03

0.025
Concentration mg g-1

0.02

0.015

0.01 Leaf
Stem
0.005

0
Control
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
Zn CU Cd Pb Cr
Treatments

Fig. 8 Total chlorophyll content in A. spinosus under heavy metal stress

Results and Discussion

Bio-organics

The bio-organic molecules remained high in the leaves of the control plant and exhibited a
distribution pattern of leaf>stem>root. The concentration of bio-organics such as total
soluble sugars, total protein, lipids, phenol, amino acids and photosynthetic pigments

Carotnoid content in Amaranthus spinosus

under heavy metal stress
0.9
0.8
Concentration mg g-1

0.7
0.6
0.5
0.4
0.3 Leaf
0.2 Stem
0.1
0
Control
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3
T1
T2
T3

Zn CU Cd Pb Cr
Treatments

Fig. 9 Carotenoid content in A. spinosus under heavy metal stress

1556 Appl Biochem Biotechnol (2012) 167:1550–1559

Phytoaccumulation of Zinc in

Concentration mg Kg -1
Amarathus spinosus
500
400
300
Leaf
200
Stem
100
0 Root
Control 200 mM (T1) 280 mM (T2) 350 mM (T3)

Treatments

Fig. 10 Phytoaccumulation of zinc in A. spinosus

(Figs. 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10) exhibited profound variation in response to the

accumulation of heavy metals viz. copper, zinc, lead, cadmium and chromium.
The concentration of sugar exhibited increase in the plant parts of A. spinosus on
treatment with varying concentrations of essential elements like copper and zinc, while a
decrease was observed with toxic elements. The distribution of photosynthetic pigments
displayed variation not only in response to concentration of treatment but among bioparts.
Essential elements like copper and zinc enhanced chlorophyll a, total chlorophyll and
chlorophyll a to b ratio in leaves. Zinc caused an increase in carotenoid content at lower
concentration. Nonessential elements like lead and chromium enhanced almost all parame-
ters of pigment profile in the leaves of A. spinosus as compared to the control indicative of
the tolerance of the weed to these metals. Reports suggested that high amounts of chromium
accumulation in plant tissues resulted in significant inhibition in chlorophyll, protein content
and in vitro nitrate reductase activity [16]. Heavy metals like cadmium, lead and nickel were
reported to reduce the accumulation of chlorophyll in angiosperm plants [17], but in
Amaranthus, lead and chromium enhanced pigment production.

Heavy Metals

Zinc Accumulation of zinc in the leaves increased with increasing soil concentration
(Fig. 10). The phytoaccumulation of zinc ranged from 82 to 234, 64 to 348 and 78 to
364 mg kg−1 in the roots, stem and leaves, respectively.

Phytoaccumulation of copper in
Concentration mg Kg-1

Amarathus spinosus
200
150
Leaf
100
Stem
50
0 Root
Control 60mM (T1) 200mM(T2) 300 mM
(T3)

Treatments

Fig. 11 Phytoaccumulation of copper in A. spinosus

Appl Biochem Biotechnol (2012) 167:1550–1559 1557

Phytoaccumulation of Cadmium in
Amarathus spinosus

Concentration mg Kg-1
40

30

20 Leaf

10 Stem
Root
0
Control 20 mM (T1) 25 mM (T2) 30 mM (T3)

Treatments

Fig. 12 Phytoaccumulation of cadmium in A. spinosus

Copper The phytoaccumulation of copper by A. spinosus (Fig. 11) exhibited variation

among plant parts and concentration of treatment. Among the plant parts, the roots accu-
mulated more copper than the stem and leaf. A steady increase was noticed in the accumu-
lation of enhancing the copper concentration in the soil. The roots accumulated copper
ranging from 93.33 to 143.33 mg kg−1 while the stem from 23.33 to 133.33 mg kg−1 and leaf
from 49.99 to 79.99 mg kg−1.

Cadmium The cadmium content in the control plants was below detectable levels (Fig. 12).
Phytoaccumulation of cadmium in the roots (20 to 34 mg kg−1) was high compared to the
leaf (8 to 20 mg kg−1) and stem (14 to 27 mg kg−1).

Lead Accumulation of lead was high in the roots followed by the stem and leaves (Fig. 13).
The concentration of lead ranged from 43.33 to 86.66, 86.66 to 126.66 and 50.11 to
181.66 mg kg−1 in the roots, stem and leaves, respectively.

Chromium The concentration of chromium was high in the roots followed by the stem and
leaves (Fig. 14). The bioassay revealed that the concentration of chromium varied from 30 to
22, 15 to 22 and 18 to 28 mg kg−1 in the leaves, stem and roots, respectively.

Bioconcentration Factor and Translocation Factor BCF is the ratio of the metal concen-
tration found within the tissues over the metal concentration found in the soil. The greater is
the coefficient, the greater will be the uptake of heavy metal. The values computed herein

Phytoaccumulation of Lead in
Amarathus spinosus
Concentration mg Kg-1

250
200
150
Leaf
100
Stem
50
Root
0
Control 30 mM (T1) 90 mM (T2) 180 mM (T3)

Treatments

Fig. 13 Phytoaccumulation of lead in A. spinosus

1558 Appl Biochem Biotechnol (2012) 167:1550–1559

Phytoaccumulation of Chromium
in Amarathus spinosus

Concentration mg Kg-1
80

60

40 Leaf

20 Stem
Root
0
Control 60 mM (T1) 180 mM (T3) 360 mM (T3)

Treatments

Fig. 14 Phytoaccumulation of chromium in A. spinosus

were above unity in most of the treatments and increased as the concentration of treatment
increased, indicating the metal accumulation capability of the weed. BCF in the bioparts
under chromium stress displayed decrease as the concentration of treatment increased, while
an increase was recorded under cadmium stress.
TF is indicative of the ability of the plant to translocate metal from the roots to shoot. A
TF value greater than 1 is indicative of metal accumulation and transport into the different
plant parts, and less than 1 is suggestive of storage of metal in roots. TF values of lead,
cadmium and zinc were above unity in Amaranthus, indicative of translocation and storage
in aerial parts. TF values of copper reached unity at higher concentration. Copper and
cadmium were accumulated in the roots at lower concentration of treatment, while translo-
cation became efficient at higher concentration. Chromium remained in the roots as treat-
ment concentration increased. Plants with both bioconcentration factor and translocation
factor greater than 1 (TF and BCF >1) have the potential to be used in phytoextraction.
Besides, plants with bioconcentration factor greater than 1 and translocation factor less than
1 (BCF >1 and TF <1) have the potential for phytostabilization [18]. A few plant species
have been reported to accumulate metal to high concentrations in the above-ground parts and
are called hyperaccumulators [19]. A. spinosus was not a hyperaccumulator of metals as it
accumulated metals less than 1,000 mg kg−1, but it exhibited phytoaccumulation efficacy.
Restriction of upward movement from roots into shoots can be considered as one of the
tolerance mechanism [20] (Table 1).
In general, the concentration of bio-organic molecules such as total soluble sugars, total
protein, lipids, phenol, amino acids and photosynthetic pigments exhibited profound varia-
tion in response to the accumulation of heavy metals which is in accordance with reports that
explain the effect of uptake of cadmium on plant metabolism through reduction in the

Table 1 Bioaccumulation factor and translocation factor in A. spinosus

Bioparts Copper (mM) Zinc (mM) Lead (mM) Cadmium (mM) Chromium (mM)

60 200 300 200 280 300 30 90 180 20 25 30 60 180 360

Leaf 0.57 0.52 0.55 0.60 0.85 2.15 0.51 0.58 0.68 2.67 2.00 4.00 1.33 0.68 0.65
Stem 0.26 0.27 0.91 0.50 0.59 2.06 1.00 1.09 1.07 4.30 5.00 5.80 1.83 1.36 0.31
Root 1.08 1.01 0.98 0.64 1.60 1.12 0.58 1.00 0.70 6.10 4.10 6.10 0.60 0.40 0.35
TF 0.78 0.79 1.41 1.73 1.21 3.04 2.5 1.65 2.4 1.10 1.45 1.44 4.70 5.10 2.64
Appl Biochem Biotechnol (2012) 167:1550–1559 1559

biomass, chlorophyll, protein, soluble sugars and amino acids [21]. Accumulation of copper,
lead and cadmium was high in the roots followed by the stem and leaves of A. spinosus.
Accumulation of zinc and chromium remained high in the aerial parts exhibiting efficient
translocation. A steady increase was noticed in the bioaccumulation of copper, zinc and
cadmium on enhancing the concentration of the corresponding metal in the soil. The present
investigation emphasized the capability of A. spinosus in heavy metal accumulation and its
effective role in pollution abatement in an eco-friendly and cost-effective manner, in turn
leading to the sustainable utilisation of the weed.

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