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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, Iranb Department of Chemical and Petroleum Engineering, Sharif University of Technology, Tehran, Iranc Department of Environmental Management, Science and Research Branch, Islamic Azad University, Tehran, Iran

a r t i c l e i n f o

Article history:Received 29 May 2011

Received in revised form

16 July 2011

Accepted 21 July 2011Available online 11 August 2011

Keywords:

Phytoremediation

Cs accumulation

Calendula alata

Amaranthus chlorostachys

Chenopodium album

a b s t r a c t

Uptake rate of 133Cs, at three different concentrations of CsCl, by Calendula alata, Amaranthuschlorostachys 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 kg1 at a 133Cs supply level

of 3.95 mg l1 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.

1. Introduction

There has been significant 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 confine a significant portion of the

market for waste management. A lot of plant species have been

identified 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 concen-

trations in plant shoots (Alloway et al., 1990; Tanhan et al., 2007).

Plants uptake system is defined as the system that involves in ionsuptake. The main components of the species-specific 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 signifi-

cantly diversified (Verkleij et al., 2009). Moreover plant concen-

trations of metals may be influenced by a variety of conditions.

Not only pH but also other ions concentrations and environmental

conditions may interact with uptake of elements and sometimeschange 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 field 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 distribu-

tion in plants and the movement of this element in ecosystems

extends back to the 1950s by the development of nucleartechnologies 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 volatiliza-

tion, release and dispersal in major reactor accidents, and the great

quantity and persistence of 137Cs in spent fuel and reprocessed

wastes (Pipıska 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 find any

differences between the uptake of radioactive and stable Cs

isotopes by Helianthus annuus L. Stable Cs is phytotoxic in solution

Contents lists available at ScienceDirect

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

Ecotoxicology and Environmental Safety

0147-6513/$- see front matter & 2011 Elsevier Inc. All rights reserved.

doi:10.1016/j.ecoenv.2011.07.019

n Corresponding author. Fax: þ 98 21 77902820.

E-mail addresses: [email protected],

[email protected] (R. Moogouei).

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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 semi-

arid regions of the world including Iran. There is no report on theuse 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 vermiculite3:1 (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 l1); 600 ml

KNO3 (107 g l1); 400 ml Ca (NO3)2 4H2O (236 g l1); 200 ml MgSO4 7H2O

(246 g l1); 150 ml Fe-EDTA (5 g l1). The composition of micro elements

(100 ml of solution all together) also was: H3BO3 (0.38 g l1); ZnSO4 7H20

(0.22 g l1); MnSO4 4H2O (1.02 g l1); CuSO4 5H2O (0.08 g l1); (NH4)6Mo7O24

4H2O (0.02 g l1). 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 flasks (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 l1 of CsCl, respectively. Consequently

the concentration of Cs ions in the solutions was 0.47, 1.58 and 3.95 mgl1. 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 flasks (Singh

et al., 2009). pH of the solution was adjusted to 5.5. The average root lengths were

300 mm (Calendula alata), 350 m m ( Amaranthus chlorostachys) and 200 mm

(Chenopodium album). Those for plants shoots were 350 mm (Calendula alata),

400mm ( Amaranthus chlorostachys) and 300 mm (Chenopodium album). Calendula alata

and Amaranthus chlorostachys plants had a massive root system. Each flask contained

three plants, which represented one replicate. Plants grown in water served as controlsamples. 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

% uptake ¼ ½ðC 0C 1Þ=C 0 100

where C 0 and C 1 are initial and remaining concentrations of metal, respectively, in

solution (mg l1) (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

flame atomic absorption spectrophotometry. The concentrations of elements in

the samples are reported on a dry matter basis.

2.2.3. Concentration ratio

The concentration ratio (CR), defined 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 confirm

the variability of results, all the data were subjected to analysis of variance to

consider the significance 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 efficient 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 significantly higher Cs concentrations in the shoots of

Calendula alata, Chenopodium album and Amaranthus chlorostachys.

Amaranthus chlorostachys showed a significant accumulation of Cs.

This plant accumulated significantly more Cs in shoots than the

other plants in all three treatments, and in treatment 1 (0.50 mg1

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 (concen-

tration 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 l1. 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 replicates7SD. po0.05, data

differences are significant.

Plant In initial

solution (mg l1)

After 15 days

(mg l1)

Remediation

(%)

Calendula alata 0.47 0.2570.01 4672.12

Calendula alata 1.58 0.9270.02 4171.59

Calendula alata 3.95 1.8970.04 5271.02

Chenopodium album 0.47 0.1570.01 6872.12



Amaranthus chlorostachys 0.47 0.2570.04 4578.59



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concentrations in solution, and the metal accumulation in shoots

was always significantly 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 l1). Moreover, the study

performed by Lasat et al. (1998) indicated Amaranthus retroflexus

accumulated high level of 137Cs in shoots. Calendula alata and

Amaranthus chlorostachys accumulated Cs above 1000 mg kg1 in

the shoots (Baker and Brooks, 1989; Yoon et al., 2006) but Csaccumulation in shoots of Chenopodium album was less than

1000 mg kg1. 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)

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

plantlets were exposed to 137Cs solution (5 103 kBql1) 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 plant’s potential for phytoremediation purpose (Yoon

et al., 2006). The process of phytoremediation action generally

requires the translocation of metals to the easily harvestable plantparts. 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/water–root

and root–shoot transfers, and therefore have much less accumula-

tion 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 efficient

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.

5. Conclusion

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 l1) Chenopodium album and at 1.58 mg l1

Cs solution Amaranthus chlorostachys were considered as the most

promising species for phytoremediation of Cs. The results present

0

10

20

30

40

50

60

70

80

0.47

( % )

mg l-1

C. alata

C. album

A. chlorostachys

1.58 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 threereplicates7SD. (Initial Cs concentrations were 0.47, 1.58 and 3.95 mg l1)

po0.05, data differences are significant.

0

500

1000

1500

2000

2500

Root

m g k g - 1 d . w .

Control T1 T2 T3

C. alataC. album

A. chlorostachys

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 l1 in Cs solution; T2: 1.58 mg l1

in Cs solution; T3: 3.95 mg l1 in Cs solution. Control values are not detectable. Po0.05 data differences are significant.

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 Chenopodium album Amaranthus chlorostachys

Cs (mg l1) 0.47 1.58 3.95 0.47 1.58 3.95 0.47 1.58 3.95

CR 1.7670.01 1.0770.03 2.1970.05 1.0670.05 1.1970.02 3.3370.03 0.7470.04 2.8470.06 4.8970.04

Concentration ratios (CR) are expressed by the means7standard deviations of 3 replicates. Po0.05 data differences are significant.

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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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