Ecotoxicology and Environmental Safety 74 (2011) 20362039

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Ecotoxicology and Environmental Safety

journal homepage: www.elsevier.com/locate/ecoenv

Phytoremediation of stable Cs from solutions by Calendula alata, Amaranthus

chlorostachys and Chenopodium album
Roxana Moogouei a,n, Mehdi Borghei b, Reza Arjmandi c
a

Department of Environmental Engineering, North Tehran Branch, Islamic Azad University, Tehran, Iran
Department of Chemical and Petroleum Engineering, Sharif University of Technology, Tehran, Iran
c
Department of Environmental Management, Science and Research Branch, Islamic Azad University, Tehran, Iran
b

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 29 May 2011
Received in revised form
16 July 2011
Accepted 21 July 2011
Available online 11 August 2011

Uptake rate of 133Cs, at three different concentrations of CsCl, by Calendula alata, Amaranthus
chlorostachys and Chenopodium album plants grown outdoors was studied. These plants grow
abundantly in semi-arid regions and their varieties exist in many parts of the world. When exposed
to lowest Cs concentration 68 percent Cs was remediated by Chenopodium album. 133Cs accumulation in
shoots of Amaranthus chlorostachys reached its highest value of 2146.2 mg kg  1 at a 133Cs supply level
of 3.95 mg l  1 of feed solution. The highest concentration ratio value was 4.89 for Amaranthus
chlorostachys, whereas for the other tests it ranged from 0.74 to 3.33. Furthermore uptake of 133Cs by
all three species increased with increasing metal concentrations. The results also indicated that
hydroponically grown Calendula alata, Amaranthus chlorostachys and Chenopodium album could be used
as potential candidate plants for phytoremediation of solutions contaminated with Cs.
& 2011 Elsevier Inc. All rights reserved.

Keywords:
Phytoremediation
Cs accumulation
Calendula alata
Amaranthus chlorostachys
Chenopodium album

1. Introduction
There has been signicant interest in the use of plants for
remediation of environmental contamination (Meyers et al., 2008).
In recent years several preliminary cost effective analyses launch
that phytoremediation should conne a signicant portion of the
market for waste management. A lot of plant species have been
identied as hyperaccumulators, i.e., they have the ability to
accumulate high concentrations of metals, without impact on
their growth and development (Xiong, 1997). Many studies have
examined the ability of plants to remediate a variety of elements
from diverse media. The achievement of phytoremediation
depends on plant growth rate and obtaining high metal concentrations in plant shoots (Alloway et al., 1990; Tanhan et al., 2007).
Plants uptake system is dened as the system that involves in ions
uptake. The main components of the species-specic uptake
system are transporters and channels (Maestri et al., 2010).
Depending on the plants uptake system, and following organ
distribution of elements, their content and distribution is signicantly diversied (Verkleij et al., 2009). Moreover plant concentrations of metals may be inuenced by a variety of conditions.
Not only pH but also other ions concentrations and environmental

Corresponding author. Fax: 98 21 77902820.

E-mail addresses: r_moogoui@iau-tnb.ac.ir,
moogouei_roxana@yahoo.com (R. Moogouei).
0147-6513/$ - see front matter & 2011 Elsevier Inc. All rights reserved.
doi:10.1016/j.ecoenv.2011.07.019

conditions may interact with uptake of elements and sometimes

change the growth rate of plants (Massas et al., 2010). There has
been an enduring interest in selecting native plants that are
tolerant to pollutants and many studies have evaluated the
phytoremediation potential of native plants under eld conditions
(McGrath and Zhao, 2003). Experimental real life studies are
necessary and may have to include a range of contaminant
concentrations, mixtures of various contaminants, and different
experimental treatments. Plant selection is based on growth rate,
contaminant translocation, accumulation potential and tolerance
to contaminants. Singh et al. (2009) have found that plants
belonging to Chenopodiaceae, Amaranthaceae and Asteraceae
families are effective remediators of 137Cs. Interest in Cs distribution in plants and the movement of this element in ecosystems
extends back to the 1950s by the development of nuclear
technologies used for energy production (Cook et al., 2007). The
radioisotopes of cesium (134Cs and 137Cs) may be of special
concern because of their similar behavior to the necessary element
K in plants, their solubility in aquatic ecosystems, the volatilization, release and dispersal in major reactor accidents, and the great
quantity and persistence of 137Cs in spent fuel and reprocessed
wastes (Pipska et al., 2004; Pinder III et al., 2006). According to
studies carried out by Tsukada et al. (2002) and Vinichuk et al.
(2010) strong correlations exist between distribution of 137Cs and
stable Cs in plants. Moreover Soudek et al. (2006) did not nd any
differences between the uptake of radioactive and stable Cs
isotopes by Helianthus annuus L. Stable Cs is phytotoxic in solution

R. Moogouei et al. / Ecotoxicology and Environmental Safety 74 (2011) 20362039

culture exceeding 200 mM (Borghei et al., 2011). The main purpose

of this work has been largely to evaluate the potential of
Chenopodium album, Amaranthus chlorostachys and Calendula alata
(Borghei et al., 2011) to phytoremediate stable cesium as an
analog of radioactive Cs. Moreover metal concentration in shoots
of plants was compared with those in roots. Chenopodium album,
Amaranthus chlorostachys and Calendula alata are fast growing
plants that grow on wide geographical locations in arid and semiarid regions of the world including Iran. There is no report on the
use of these plants for phytoremediation of stable Cs from
solutions as well as nuclear waste.
2. Materials and methods
Three plant species (Amaranthus chlorostachys var. Chlorostachys, Calendula
alata Rech. F., Fl. Iranica and Chenopodium album) were used in this study with the
aims to evaluate their potential for phytoremediation of Cs solutions and their
tolerance to Cs.
2.1. Plant material and hydroponics culture
Healthy seeds of Amaranthus chlorostachys, Calendula alata and Chenopodium
album were surface sterilized by one percent sodium hypochlorite for 20 min.
Calendula alata seeds were sown in a substrate containing perlite and vermiculite
3:1 (v/v) moistened with distilled water for four weeks until seedlings with two
leaf pairs were established. Amaranthus chlororstachys and Chenopodium album
seeds were germinated in sand. Then one-month-old plantlets were transplanted
in plastic trays containing 10 L nutrient solutions. The composition of macro
elements per 100 L solution was as follows: 100 ml NH4H2PO4 (115 g l  1); 600 ml
KNO3 (107 g l  1); 400 ml Ca (NO3)2  4H2O (236 g l  1); 200 ml MgSO4  7H2O
(246 g l  1); 150 ml Fe-EDTA (5 g l  1). The composition of micro elements
(100 ml of solution all together) also was: H3BO3 (0.38 g l  1); ZnSO4  7H20
(0.22 g l  1); MnSO4  4H2O (1.02 g l  1); CuSO4  5H2O (0.08 g l  1); (NH4)6Mo7O24 
4H2O (0.02 g l  1). Solution pH was adjusted to 5.5 through 5.8 with 0.1 M NaOH
or 0.1 M HNO3 and continuously aerated with a pump (Islam et al., 2007). Nutrient
solutions renewed at every 10th day. Level of the solutions in trays was made up
with nutrient solution when required. Each tray contained 24 plants. Plants were
grown outdoors with temperature ranging from 31 1C through 40 1C (maximum
daily temperature) and 17 1C through 28 1C (minimum daily temperature) with
natural light during the experiment. After four weeks plantlets with uniform size
were selected and transferred to 1 L asks (Singh et al., 2009).
2.2. Experiments using hydroponically grown plants
2.2.1. Remediation of Cs solutions contaminated with Cs
The roots of cultured plants were washed thoroughly with distilled water and
plants were incubated with roots immersed in 1 L solution with three different
Cs concentrations. The treatment samples included: (1) control sample free of Cs,
samples 2, 3 and 4 containing 0.5, 2 and 5 mg l  1 of CsCl, respectively. Consequently
the concentration of Cs  ions in the solutions was 0.47, 1.58 and 3.95 mgl  1. The
experiment was arranged with each treatment in triplicate samples. The treatment
group was exposed to CsCl solution for a period of 15 days in 1500 ml asks (Singh
et al., 2009). pH of the solution was adjusted to 5.5. The average root lengths were
300 mm (Calendula alata), 350 mm (Amaranthus chlorostachys) and 200 mm
(Chenopodium album). Those for plants shoots were 350 mm (Calendula alata),
400 mm (Amaranthus chlorostachys) and 300 mm (Chenopodium album). Calendula alata
and Amaranthus chlorostachys plants had a massive root system. Each ask contained
three plants, which represented one replicate. Plants grown in water served as control
samples. Distilled water was used for solution preparation and for make-up of lost
water. After treatment period samples of solutions were drawn out from the solutions
and analyzed for Cs concentrations. In all experiments Cs contents of solutions were
determined using atomic absorption spectrophotometry (Variance Spectra AA-55B).
The percentage metal uptake was calculated from

2.2.3. Concentration ratio

The concentration ratio (CR), dened as the ratio of metal concentrations in
plant shoots to those in the roots (Gonzaga et al., 2006; Bidar et al., 2007) was
calculated to check the effectiveness of plants in translocating metals to their
aerial parts (Dahmani-Muller, et al., 2000; Zab"udowska et al., 2009).
2.2.4. Statistical analysis
The experiments were performed in triplicate and the statistical analysis was
performed using Statistical Analysis System (SAS) software package. To conrm
the variability of results, all the data were subjected to analysis of variance to
consider the signicance differences. Moreover means comparison between data
was obtain using Duncan test.

3. Result
3.1. Cs remediation from solutions using hydroponically
grown plants
In the present study, the plants were found to be efcient in
remediating solutions contaminated with Cs (Table 1). As it is
presented in Fig. 1 when these plants were exposed to lowest Cs
concentration 68 percent Cs was remediated by Chenopodium
album that was the highest remediation percentage found in
this study.
3.2. Cs concentration in plants
As it is presented in Fig. 2, increased amounts of Cs in solutions
led to signicantly higher Cs concentrations in the shoots of
Calendula alata, Chenopodium album and Amaranthus chlorostachys.
Amaranthus chlorostachys showed a signicant accumulation of Cs.
This plant accumulated signicantly more Cs in shoots than the
other plants in all three treatments, and in treatment 1 (0.50 mg  1
CsCl) Calendula alata roots showed the lowest Cs concentration.
In this study all treated plants continued to produce new organs.
3.3. Cs concentration ratios in plants
Both the highest level of Cs accumulation and the highest
ability to translocate it from roots to shoots were observed in
Amaranthus chlorostachys. As shown in Table 2, the CR (concentration ratio) value was 4.89 for Amaranthus chlorostachys,
whereas for the other tests it ranged from 0.74 to 3.33.

4. Discussion
In this study, the percentage uptake of Cs was highest for
Chenopodium album at a Cs supply levels of 0.47 mg l  1. In general,
the metal accumulation in Calendula alata, Chenopodium album
and Amaranthus chlorostachys increased with the increase in metal

Table 1
Cs concentrations. All the values are means of three replicates 7 SD. p o 0.05, data
differences are signicant.
Plant

In initial
solution (mg l  1)

After 15 days
(mg l  1)

Remediation
(%)

Calendula alata
Calendula alata
Calendula alata
Chenopodium album
Chenopodium album
Chenopodium album
Amaranthus chlorostachys
Amaranthus chlorostachys
Amaranthus chlorostachys

0.47
1.58
3.95
0.47
1.58
3.95
0.47
1.58
3.95

0.257 0.01
0.927 0.02
1.89 7 0.04
0.157 0.01
0.957 0.05
1.85 7 0.29
0.257 0.04
0.547 0.06
2.307 0.15

46 72.12
41 71.59
52 71.02
68 72.12
39 73.48
52 77.57
45 78.59
65 74.11
41 73.92

% uptake C 0 C 1 =C 0   100
where C0 and C1 are initial and remaining concentrations of metal, respectively, in
solution (mg l  1) (Abdel-Halim et al., 2003; Tanhan et al., 2007).
2.2.2. Distribution of Cs in Calendula alata, Amaranthus chlorostachys
and Chenopodium album
At the end of the experiment, plants were thoroughly washed with distilled
water, separated into root and shoot and dried in an oven at 60 1C for 48 h. The
dried samples were digested in HNO3:HClO4 (5:1, V/V) and analyzed for Cs by
ame atomic absorption spectrophotometry. The concentrations of elements in
the samples are reported on a dry matter basis.

2037

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R. Moogouei et al. / Ecotoxicology and Environmental Safety 74 (2011) 20362039

concentrations in solution, and the metal accumulation in shoots

was always signicantly higher than those in roots for all three
species. In the present study Amaranthus chlorostachys showed the
highest percentage uptake of Cs in shoots at the highest Cs
concentration in solution (3.95 mg l  1). Moreover, the study
performed by Lasat et al. (1998) indicated Amaranthus retroexus
accumulated high level of 137Cs in shoots. Calendula alata and
Amaranthus chlorostachys accumulated Cs above 1000 mg kg  1 in
the shoots (Baker and Brooks, 1989; Yoon et al., 2006) but Cs
accumulation in shoots of Chenopodium album was less than
1000 mg kg  1. The shoots Cs concentration were much greater
than those of the roots Cs contents, indicating high mobility of Cs
from the roots to the shoots of all three species while no symptom
of immobilization of Cs was observed in roots. Singh et al. (2008)

80
70

C. alata
C. album
A. chlorostachys

60

(%)

50
40

have found that when Vetiver grass (Vetiveria zizanoides) L. Nash

plantlets were exposed to 137Cs solution (5  103 kBql  1) 137Cs
accumulation occurred more in roots than shoots. Restriction of
upward movement of metals from roots into shoots can be
considered as one of the tolerance mechanism (Verkleij and
Schat, 1990). Moreover concentration ratio (CR) can be used to
estimate a plants potential for phytoremediation purpose (Yoon
et al., 2006). The process of phytoremediation action generally
requires the translocation of metals to the easily harvestable plant
parts. Comparing CR, the ability of various plants in remediation of
metals from soils and waters and translocating them to the shoots
can be compared. Tolerant plants tend to restrict soil/waterroot
and rootshoot transfers, and therefore have much less accumulation in their biomass, while hyperaccumulators actively take up
and translocate metals into their aboveground biomass so CR
values less than one are unsuitable for phytoremediation (Fitz and
Wenzel, 2002). Therefore present results may indicate that plant
species were not tolerant of cesium and translocate this element
to their shoots. In this respect they are considered to be efcient
for Cs removal by phytoremediation. Cs concentration ratio
for Amaranthus chlorostachys was 0.7470.04 at the lowest Cs
concentration that showed two other Cs solutions with higher
concentrations are more suitable for phytoremediation of Cs
solutions. All three species in most treatment tested were capable
of accumulating and translocating Cs in the shoots.

30
20

5. Conclusion

10

In several tests Amaranthus chlorostachys, Chenopodium album

and Calendula alata had CRs greater than one so could be used as
potential candidate plants for phytoremediation of Cs from
solutions. Amaranthus chlorostachys was the most effective in
taking up cesium with CR ranging from 0.47 to 4.89. Among
these plants species at lowest and highest Cs concentrations
(0.47 and 3.95 mg l  1) Chenopodium album and at 1.58 mg l  1
Cs solution Amaranthus chlorostachys were considered as the most
promising species for phytoremediation of Cs. The results present

1.58
mg l-1

0.47

3.95

Fig. 1. Remediation percent of solutions contaminated with Cs by C. alata,

C. album and A. chlorostachys after 15 d. All the values are mean of three
replicates 7SD. (Initial Cs concentrations were 0.47, 1.58 and 3.95 mg l  1)
p o0.05, data differences are signicant.

2500
Control

T1

T2

A. chlorostachys

T3

mg kg-1 d.w.

2000
1500

C. alata

C. album

1000
500
0

Root

Shoot

Root

Shoot

Root

Shoot

Fig. 2. Cs concentration in roots and shoots of hydroponically grown plants. The values are mean of three replicates 7S.D. T1: 0.47 mg l  1 in Cs solution; T2: 1.58 mg l  1
in Cs solution; T3: 3.95 mg l  1 in Cs solution. Control values are not detectable. P o 0.05 data differences are signicant.

Table 2
Comparisons of concentration ratios (CR) in roots and shoots of Calendula alata, Chenopodium album and Amaranthus chlorostachy exposed to different solutions
contaminated with Cs for a period of 15 days.
Calendula alata
Cs (mg l  1)
CR

0.47
1.76 7 0.01

Chenopodium album
1.58
1.07 70.03

3.95
2.197 0.05

0.47
1.06 7 0.05

1.58
1.19 70.02

Amaranthus chlorostachys
3.95
3.337 0.03

0.47
0.74 7 0.04

Concentration ratios (CR) are expressed by the means7 standard deviations of 3 replicates. P o0.05 data differences are signicant.

1.58
2.84 70.06

3.95
4.89 70.04

R. Moogouei et al. / Ecotoxicology and Environmental Safety 74 (2011) 20362039

a simple yet effective method for remediation of polluted sites by

natural means.

Acknowledgments
The authors would like to express their sincere thanks to
Mrs. Sh. Teymoori and Mr. S. Moogouei for their invaluable technical
assistance. Funds and support for this work were provided by the
Morad Abad Center for Phytoremediation Research (MCPR).
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