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Chemosphere 70 (2008) 531–537

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Technical Note

Hydroponic phytoremediation of Cd, Cr, Ni, As, and Fe: Can

Helianthus annuus hyperaccumulate multiple heavy metals?
Mary C. January a, Teresa J. Cutright a,*
, Harry Van Keulen b, Robert Wei c

a
Department Civil Engineering, The University of Akron, Akron, OH 44325-3905, United States
b
Department Biological, Geological and Environmental Sciences, Cleveland State University, Cleveland, OH 44115, United States
c
Department of Chemistry, Cleveland State University, Cleveland, OH 44115, United States

Received 1 March 2007; received in revised form 21 June 2007; accepted 26 June 2007
Available online 13 August 2007

Abstract

Sundance sunflowers were subjected to contaminated solutions containing 3, 4, or 5 heavy metals, with and without EDTA. The sun-
flowers exhibited a metal uptake preference of Cd = Cr > Ni, Cr > Cd > Ni > As and Fe >> As > Cd > Ni > Cr without EDTA and
Cr > Cd > Ni, Fe >> As > Cd > Cr > Ni with EDTA. As uptake was not affected by other metals, but it decreased Cd and Ni concen-
tration in the stems. The presence of Fe improved the translocation of the other metals regardless of whether EDTA was present. In
general, EDTA served as a hindrance to metal uptake. For the experiment with all five heavy metals, EDTA decreased Cd in the roots
and stems from 2.11 to 1.36 and from 2.83 to 2.32 mg g1 biomass, respectively. For the same conditions, Ni in the stems decreased from
1.98 to 0.94 mg g1 total metal uptake decreased from 14.95 mg to 13.89 mg, and total biomass decreased from 2.38 g to 1.99 g. These
results showed an overall negative effect in addition of EDTA. However it is unknown whether the negative effect was due to toxicity
posed by EDTA or the breaking of phytochelatin-metal bonds. The most important finding was the ability of Sundance sunflowers
to achieve hyperaccumulator status for both As and Cd under all conditions studied. Ni hyperaccumulator status was only achieved
in the presence of three metals without EDTA.
Ó 2007 Elsevier Ltd. All rights reserved.

Keywords: Helianthus annuus hyperaccumulator; Sunflowers; Heavy metals; Phytoremediation

1. Introduction been reported in the mining districts of Spain, agricultural

There are a total of 31 metal contaminated sites in Ohio;
25 of which are on the National Priority List (OEPA,
2005). Of the most heavily contaminated sites, 25 contain
arsenic, 20 have chromium, 19 have cadmium, 16 contain
nickel, and 13 sites are contaminated with all four metals
(OEPA, 2005). The Ohio sites contain similar contaminant
characteristics as heavy metal sites all over the world. For
instance, more than 1.4% of the total soil surface in Flan-
ders Belgium is heavily contaminated with heavy metals
(Meers et al., 2005). Toxic levels of heavy metals have also
*
Corresponding author. Tel.: +1 330 972 4935; fax: +1 330 972 6020.
E-mail address: tcutright@uakron.edu (T.J. Cutright).
fields inn China (Zhang et al., 2005), industrial sections of
Germany (Evangelou et al., 2006) and in Uttar Pradesh
India (Gupta and Sinha, 2007).
Phytoremediation is the use of green plants to remove
contaminants from soil and groundwater or to lower con-
taminant mobility (Madrid et al., 2003). It has become a
promising remediation technique with the discovery of
hyperaccumulators; plants that are able to take up large
quantities of metals (Roosens et al., 2003). Achievement
of hyperaccumulator status is based on the ability to
uptake and retain within the shoot (stem and leaves) one
of the following metals at a minimum of: 10 mg g1 of
Zn or Mn, 1 mg g1 of Ni, Co, Cr, Cu or Pb, 0.1 mg g1
of As or Cd (Roosens et al., 2003; Turgut et al., 2005).
Different strains of the Brassica, Alyssum, Arabidopis,

0045-6535/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.chemosphere.2007.06.066
532 M.C. January et al. / Chemosphere 70 (2008) 531–537
and Petrisis species have been the most widely studied of
the more than 400 hyperaccumulating vascular plants
(Roosens et al., 2003). It is important to note that most
of the known hyperaccumulators are selective toward one
metal and would not be effective at sites with multiple
heavy metals (Kamnev and van der Lelie, 2000).
One approach to increase the uptake of heavy metals is
the use of chelators to mobilize the metals to the root zone
(Madrid et al., 2003). The viability of this approach is well
documented, however the effect of chelators on the plants
themselves has not been critically examined. The most
commonly used and effective chelator is EDTA (Wu
et al., 2004; Liphadzi and Kirkham, 2006; Ruley et al.,
2006). Although effective at mobilizing metals, EDTA is
poorly biodegraded in the soils (Luo et al., 2006). EDTA
also poses several disadvantages. It can decrease plant bio-
mass, destroy the physiological barriers of the root, or
inactivate transporter proteins to the extent that its metal
mobilizing and translocation benefits are minimized (do
Nacimento et al., 2006; Liphadzi and Kirkham, 2006;
Luo et al., 2006). Non-synthetic chelators such as citric
acid and EDDS (S,S-(N,N 0 )-ethylenediaminodisuccinic
acid, an analogue of EDTA) are initially comparable at
mobilizing metals, but do not provide long term effective-
ness since they are easily biodegraded (Turgut et al.,
2005; do Nacimento et al., 2006; Evangelou et al., 2006).
The objective of this research was to investigate the
effectiveness of Helianthus annuus for the phytoremediation
of mixed metals. A key goal was to ascertain if H. annuus
could hyperaccumulate more than one heavy metal. The
experiments described in this manuscript were conducted
under hydroponic conditions, to eliminate the mass trans-
fer (bioavailability) issues that would be present in soil
experiments. Removing mass transfer limitations via hyd-
proponics is critical for elucidating free metal ion and
metal-chelate uptake and translocation within the plant
(Marchiol et al., 2004; Cosio et al., 2006; Hernandez-Allica
et al., 2007). Although unnecessary in a hydroponic envi-
ronment, EDTA was used in some experiments in order
to examine its effects on metal uptake, translocation and
total plant biomass.
2. Materials and methods
2.1. Heavy metal and chelator: source and concentration
Sunflowers were exposed to a solution containing a mix-
ture of heavy metals. Each heavy metal was added at
30 mg l1, where the 30 mg l1 refers to the concentration
of the individual metal, not the compound added. The
metals were applied as As5, (Na2HAsO4 Æ 7H2O), Cd2+
(CdSO4 Æ 8H2O), Cr3+ (CrCl3 Æ 6H2O), Ni2+ (NiSO4 Æ
6H2O) and Fe (FeSO4 Æ 7H2O). EDTA was added at
0.265 g l1 (0.708 mM) to approximate a concentration of
0.1 g kg1 in soil applications (data not shown) to evaluate
the impact of synthetic chelators on metal uptake and
translocation.
2.2. Cultivar source and seedling preparation
The Sundance cultivar of the dwarf sunflower, H.
annuus, was used for each hydroponic experiment. The
seeds were initially grown in Rockwool in a greenhouse
illuminated with natural light. The average greenhouse
temperature was 28 oC (winter) or 35 oC (summer) in the
day and 20 oC at night. Seedlings were allowed to grow
for 4 wk using a nutrient solution containing 250 mg N
(NH4NO3), 109 mg P (KH2PO4) and 207 mg K (KH2PO4)
per 1 l distilled water. After the growth period, seedlings
of similar size were transferred to troughs in their indi-
vidual Rockwool compartment to initiate the greenhouse
experiment.
2.3. Greenhouse experiment
Seven Sundance sunflowers were utilized per chamber,
with two chambers used simultaneously per experiment.
The first set of experiments investigated the impact of
EDTA on the uptake and translocation of Cd, Cr, and
Ni. One set of plants was subjected to a solution containing
30 mg l1 of Cd, Cr and Ni for a combined metal concen-
tration of 90 mg l1. The second set of plants was subjected
to a solution of 30 mg l1 Cd, Cr, Ni and 0.265 g l1
(0.708 mM) EDTA. Both solutions were introduced at a
complete recycle rate of 1.59 l h1. After 17 d of exposure,
the plants were harvested for analysis.
The second set of experiments investigated the impact of
As. One set of plants was subjected to a solution containing
30 mg l1 As as the sole contaminant; the second set
received a mixed solution containing 30 mg l1 of As, Cd,
Cr and Ni. EDTA was not utilized during this experiment
in order to garner a better understanding as to how As
would impact translocation.
The final experimental set explored the effect of a fifth
metal and synthetic chelators on the Sundance sunflowers.
The two sets of Sundance dwarf sunflowers were exposed
to As, Cd, Cr, Fe and Ni all at 30 mg l1 for each metal.
One set of plants was also subjected to 0.265 g l1 EDTA.
The experiment was conducted for 17 d as described above.
All of the heavy metal concentrations used in the afore-
mentioned experiments were based on typical contaminant
levels in Northeast Ohio (OEPA, 2005). The EDTA con-
centration was based on previous research results with
Cd, Cr and Ni (Turgut et al., 2005).
3.1. Analytical methods
3.1.1. Determination of heavy metal content
Each biomass section was digested separately to ascer-
tain the metal content in the individual section. One g of
milled plant tissue was soaked in 20 ml of concentrated
nitric acid for 6 h. The mixture was then boiled to 50%
of the original volume. Next, 4 ml of perchloric acid was
added and the mixture was refluxed for 90 min. The solu-
 M.C. January et al. / Chemosphere 70 (2008) 531–537 533

tion was then diluted with distilled water to a total volume

a 6 Sundance
of 20 ml and analyzed by FAAS.
Buck 200 AA Spectrophotometer was used to analyze

mg Metal g-1 biomass

5
metal concentrations. The detection limits for the target
4
metals are 0.028 mg l1 for Cd, 0.078 mg l1 for Cr, Roots
Leaves
0.14 mg l1 for Ni and 0.25 mg l1 for As. The concentra- 3 Stem
tions of Cd, Cr, Ni and As were determined by calibration
2
curves obtained using standards solutions of pure metal
ions (Fisher Scientific). Calibration curves were generated 1
using six replicates per metal concentration. New standard
0
solutions were used to generate new curves every month. In
addition, blind samples that were also analyzed by an b 4
external, EPA approved lab were used for quality assur- 3.5
Roots

mg Metal g-1 biomass

Stem
ance of in-house analytical methods. 3 Leaves
2.5
3.1.2. Statistical analysis 2
The results from the four studies were analyzed using 1.5
MINITAB version 14 software. Experimental results were 1
compared using a general linear model. Statistical signifi- 0.5
cance was determined using Tukey comparisons. P-values 0
under 0.05 were considered statistically significant. Cd Cr Ni

Fig. 1. Metal concentration in biomass for Sundance exposed to 30 mg l1

4. Results and discussion each Cd, Cr, Ni. (a) No EDTA and (b) with EDTA. Error bars show
variation between replicates. All differences in sequestration location are
significant with the exception of Cd leaves-stems, Cr leaves-stems and Ni
4.1. Impact of EDTA on Cd, Cr and Ni uptake roots-stems (p = 0.0542).

It is generally believed that hyperaccumulating plants

will sequester toxic metals in the vacuoles of the plant with
the leaf vacuoles being the dominant location (Yang et al.,
2005). The effect of EDTA varied for each metal as shown
by the metal concentration in the biomass (Fig. 1). Cad-
mium concentrations in the roots were significantly (p >
0.05) increased by EDTA, while stem concentration
decreased significantly from 3.33 mg g1 biomass to
1.34 mg g1 biomass (p = 0.026). The leaves contained sim-
ilar levels of cadmium, 0.88 mg g1 biomass and
0.66 mg g1 biomass, with and without EDTA, respec-
tively. Zhang et al. (2005) had also found EDTA to be inef-
fective for Viola baoshanensis or Vertivera zizanoides.
Conversely, chromium concentration with the addition of
EDTA was higher in the stems, 1.94 mg g1 biomass
(0.66 mg g1 biomass without EDTA), and lower in the
roots, 4.88 mg g1 biomass (2.98 mg g1 root without
EDTA). This was surprising since other researchers have
found Cr uptake and translocation not to be enhanced
by EDTA (Meers et al., 2005). For nickel, EDTA increased
the root concentration from 1.00 mg g1 biomass to
2.04 mg g1 biomass (p > 0.05) and decreased the stem
concentration from 2.21 mg g1 biomass to 0.78 mg g1
biomass (p < 0.05). Liphadzi and Kirkham (2006) had
found that EDTA increased both Cd and Ni in a different
strain of H. annuus.
Since differences in biomass weight can yield an appar-
ent increase/decrease in uptake concentrations, the total
metal uptake was also determined (Table 1). When EDTA
was present, the amount of Ni and Cr accumulated was
lower than without EDTA, while Cd uptake increased
(0.66 mg without EDTA, 2.15 mg with EDTA). The metal
selectivity, based on total mg accumulated was: Cd =
Cr  Ni without EDTA and Cr > Cd > Ni with EDTA.
The change in selectivity infers that the presence of EDTA
enabled the plants to uptake more Cr. However, the
majority of Cr was retained in the roots. The translocation

Table 1
Total biomass and total metal uptake for Sundance sunflowers
Experiment EDTA (g l1) Total biomass (g) Metals (mg) Total metals (mg)
As Cd Cr Ni Fe
Cd, Cr, Ni – 7.744 – 0.66 5.76 4.14 – 10.56
Cd, Cr, Ni 0.265 2.183 – 2.15 2.36 2.07 – 6.58
As, Cd, Cr, Ni – 2.682 1.95 2.40 3.92 2.55 – 10.82
As – 2.129 1.59 1.59
As, Cd, Cr, Ni, Fe – 2.381 2.36 2.02 1.55 1.57 7.45 14.94
As, Cd, Cr, Ni, Fe 0.265 1.991 2.36 1.72 1.47 1.07 7.26 13.89
534 M.C. January et al. / Chemosphere 70 (2008) 531–537

Table 2 tion was highest in the roots at 1.54 mg g1 biomass

Translocation factors (TF) for hydroponic experiments with Sundance
sunflowers
EDTA applied As
Cd, Cr, Ni No
Cd, Cr, Ni Yes
As, Cd, Ni, Cr, No 0.4
As No 0.44
As, Cd, Cr, Ni, Fe No 0.81
As, Cd, Cr, Ni, Fe Yes 0.86
TF = shoot concentration divided by root concentration.
factors (TF) for each treatment are shown in Table 2. Com-
paring the translocation for the plants treated with EDTA
(Ni > Cr  Cd) and without EDTA (Ni  Cd  Cr) indi-
cates that the only metal effectively translocated in this
experimental set was Ni. A Ni-nicotianamine complex
was recently identified as the responsible chelator for the
translocation of Ni in Thlaspi caerulescens (Mari et al.,
2006). Other researchers have found that Ni tolerance
and transfer through the plant xylem is dependent on free
histidine. Although malic and citric acid were also effective,
free histidine translocated the largest amount of Ni (Wyc-
isk et al., 2004). In the absence of EDTA, both histidine
and nicotianamine mediated transport would explain the
increased Ni translocation. Conversely, once Ni had bound
to EDTA it was expected not to be able to complex with
histidine, thereby limiting translocation.
4.2. The impact of As on the uptake and translocation of
Cd, Cr, and Ni
Fig. 2 shows the metal concentration for each biomass
section when As was added to the contaminant solution.
When As was the only contaminant the root, stem and leaf
concentrations were 1.52, 0.52 and 1.04 mg g1 biomass,
respectively with no statistically significant selectivity.
When Cd, Cr and Ni were also present, arsenic concentra-
(0.032 > p > 0.005) and lower in the leaves and stems with
no statistical significance between leaf (0.56 mg g1) and
Cd Cr Ni Fe stem (0.79 mg g1) concentrations. The addition of Cd,
0.14 0.02 0.31 Cr and Ni did not appear to effect As uptake (p > 0.05).
0.21 0.25 0.42 In the presence of arsenic, Cd accumulation was highest
0.16 0.02 0.30
in the roots at 3.48 mg g1, and lowest in the leaves at
0.55 0.06 1.06 0.02 0.07 mg g1. The decrease in Cd stem concentration
0.31 0.07 0.76 0.04 (1.71 mg g1) may be due to competition between Cd and
As. Both Cd and As are highly thiol reactive which would
deplete the phytochelatin necessary for translocation (Li
et al., 2004). Cr concentration was highest in the roots
11.03 mg g1 (p < 0.05) and low in the leaves (0.09 mg g1)
and stems (0.33 mg g1). Nickel was also highest in the
roots at 2.47 mg g1, with 0.39 and 1.59 mg g1 in the
leaves and stems (0.002 > p > 0.0001).
Translocation of As (Table 2) was 0.44 alone and 0.40
with Cd, Cr and Ni present. The As transport through
the plant was not greatly affected by the presence of other
metals. The selectivity for translocation of metals in this
experiment was As > Ni > Cd > Cr (Table 2). Other
researchers have found that As was not readily translo-
cated to the shoots. Gulz et al. (2005) found As typically
accumulated in roots of Zea mays, Brassica napus and
some strains of H. annuus.
4.3. The impact of Fe and EDTA on the uptake and
translocation of Cd, Cr, Ni and As
The totals metals accumulated per biomass section, with
and without EDTA, are shown in Fig. 3. For this experi-
mental set, EDTA did not appear to impact As uptake in
any of the plant sections. The greatest effect of EDTA
was shown on Cd uptake. EDTA increased the Cd root
concentration from 1.36 to 2.11 mg g1 (p < 0.0001) while
reducing stem concentration from 2.83 to 2.32 mg g1 bio-
mass (p = 0.0003). There was a small but insignificant

16

14
mg Metal g-1 biomass

12 Roots
Leaves
10
Stem

0
Cd Cr Ni As As Only

Fig. 2. Metal concentration in biomass for Sundance sunflowers exposed to 30 mg l each Cd, Cr, Ni, As or only 30 mg l1 As. Error bars show variation
1

between replicates. All differences in sequestration location are significant with the exception of As (mixed) leaves-stems, Cr leaves-stems and As Only
roots-leaves-stems.
 M.C. January et al. / Chemosphere 70 (2008) 531–537
20
18
16
mg Metal per g biomass 14
12
10
8
6
4
2
0
25
20
mg Metal per g biomass
15
10
0
As
Fig. 3. Metal concentration in Sundance biomass exposed to 30 mg l each As, Cd, Cr, Ni, Fe (a) without EDTA – differences in sequestration location
are significant with the exception of As roots-leaves-stems, Cr leaves-stems and Fe leaves-stems and (b) with EDTA – differences in sequestration location
are significant with the exception of As roots-leaves and roots-stems, Cd roots-stems, Cr leaves-stems and Fe leaves-stems.
increase in Cd in the leaves from 0.05 to 0.10 mg g1.
EDTA also increased Fe uptake to the stems and leaves
from 0.71 to 0.93 mg g1 and 0.16 to 0.65 mg g1 biomass
respectively. Although these increases were not significant,
they are indicative of the formation of EDTA-Fe com-
plexes. The most significant decrease with EDTA was for
Ni in the stems, causing it to decrease from 1.98 to
0.94 mg g1 biomass (p < 0.05).
The total biomass when EDTA was used, was reduced
by 16% compared to the plants that were not exposed to
EDTA (1.99 g from 2.38 g, Table 1). The total metal
uptake was reduced from 14.95 mg to 13.89 mg with
EDTA, with an uptake selectivity of Fe  As > Cd >
Ni > Cr without EDTA and Fe  As > Cd > Cr > Ni,
with EDTA. This shows that despite the slight increase in
metal concentration in the plant tissue, the toxic effects of
EDTA reduced the actual metal uptake due to the
decreased biomass. It is not known whether the presence
of EDTA reduced biomass or the stress from increased
uptake of EDTA-metal complexes. In an earlier soil-based
study, Chen and Cutright (2001) showed a 50% and
60% decrease in shoot and root biomass of four wk old
535
Roots
Leaves
Stem
Roots
Leaves
Stem
Cd Cr Ni Fe
1
H. annuus with EDTA versus plants grown without EDTA.
In another soil study using sunflowers, Turgut et al. (2005)
demonstrated substantial decreases in plant biomass when
EDTA was added.
The TF for each treatment are shown in Table 2. The
only effective translocation shown was for Ni without
EDTA (1.06). The remaining metals were primarily located
in the roots. This most significant root contaminant was
Fe, which could have inhibited the initial binding of the
other metals to the root walls. Fe is an essential plant nutri-
ent necessary for chlorphyll synthesis. It is sometimes
added as a nutrient amendment at contaminated sites to
improve plant health by increasing the amount of chloro-
phyll (Keller and Hammer, 2004). A typical mechanism
for counteracting toxic metal levels is for the plant to gen-
erate phytosiderophores to preferentially bind more of the
metals essential for growth (Yang et al., 2005). It is possible
that in the highly toxic hydroponic environment, in which
there were no mass transfer limitations, the plants pro-
duced more phytosiderophores to preferentially uptake Fe.
EDTA did not alter As, Cd or Fe translocation (0.81 vs.
0.86, 0.06 vs. 0.07 and 0.02 vs. 0.04 with and without
536 M.C. January et al. / Chemosphere 70 (2008) 531–537
Table 3
Shoot concentrations (mg g1) for Sundance sunflowers across all
treatments. Values in bold indicate achievement of hyperaccumulator
status
Treatment (30 mg l1 each metal) Shoot concentration
Cd Cr Ni
Cd, Cr, Ni 1.03 0.4 1.07
Cd, Cr, Ni with EDTA 0.64 0.76 0.85
As
As, Cd, Cr, Ni 0.55 0.17 0.75
As, Cd, Cr, Ni, Fe 0.75 0.18 0.66
As, Cd, Cr, Ni, Fe with EDTA 0.64 0.23 0.51
EDTA, respectively). However, the presence of EDTA
reduced Cr translocation from 0.55 to 0.31 and Ni translo-
cation from 1.06 to 0.76. The selectivity for translocation
without EDTA was Ni > As > Cd  Cr > Fe and As >
Ni > Cd  Cr > Fe with EDTA, showing that EDTA
inhibited Ni translocation significantly in this case. It is
hypothesized that the presence of Fe aided the transloca-
tion of Cd and Cr as a nutrient amendment rather than
limiting uptake by reduction of Fe chelating exudates.
Additionally, As has been shown to complex with Fe and
Al (Bagga and Peterson, 2001). Thus Fe may have detoxi-
fied some of the As, allowing other metals to achieve
greater translocation within the plants.
The critical result of this project is the achievement of
the dwarf sunflower as hyperaccumulator. As stated ear-
lier, the qualifications of hyperaccumulators are the ability
to uptake more than 0.1 mg g1 As or Cd and 1.0 mg g1
Cr or Ni. Table 3 contains the shoot concentrations of
As, Cd, Cr and Ni for all experiments performed. Arsenic
hyperaccumulation was achieved in all cases. This is the
first documentation of H. annuus to hyperaccumulate As,
whether present as an individual contaminant or present
with multiple metals. Cd hyperaccumulator status was also
achieved for all experiments. The ability to hyperaccumu-
late Cd was not surprising since several strains of dwarf
sunflowers have been shown to have shoot concentrations
of Cd higher than many other plants (Murillo et al., 1999).
Cr hyperaccumulator status was not achieved in any of
the treatments indicating that dwarf sunflowers were not
effective Cr hyperaccumulators at these exposure times.
Ni hyperaccumulator status was only achieved for the
experiment where Cd, Cr and Ni was added without EDTA.
However, Ni shoot concentrations were generally higher
than Cr, indicating that with increased treatment time the
plants may be able to achieve hyperaccumulators status.
5. Conclusions
This research examined efficiency of H. annuus, dwarf
sunflowers, in the removal of multiple heavy metal contam-
inants. The efficiency of phytoremediation was based on
uptake rate (mg g1), total metal accumulated (mg), and
the sequestration location (selectivity) of individual and
mixed metals.
The addition of As to the Cd, Cr and Ni exposure served
to increase Cd within the root and decrease Cd in the stem.
This was expected since Cd and As compete for phytochel-
atins. The addition of Fe increased the Cd uptake to the
levels seen without As present. This was most likely
As achieved through the benefit of Fe as a nutrient amend-
ment. Fe addition also increased the translocation of Ni
to 1.06, without EDTA. However, As uptake and translo-
1.55
0.67
cation was not effected by the presence of Cd, Cr and Ni.
0.96 Sundance cultivars showed a metal uptake preference of
1.15 Fe  As > Cd > Ni > Cr without EDTA and Fe  As >
Cd > Cr > Ni with EDTA. As uptake and translocation
was not affected by other metals, but As decreased Cd
and Ni stem concentration. The effect of EDTA in the
hydroponic environment served as a hindrance to metal
uptake. With Fe and As present, ETDA decreased Cd in
the roots and stems from 2.11 to 1.36 and from 2.83 to
2.32 mg g1 biomass, respectively. For the same condi-
tions, Ni in the stems decreased from 1.98 to 0.94 mg g1,
total metal uptake decreased from 14.95 mg to 13.89 mg,
and total biomass decreased by 84% (2.38 g to 1.99 g). In
this case, EDTA also decreased Cd TF from 0.55 to 0.33
and Ni from 1.06 to 0.76. These results showed an overall
negative effect in addition of EDTA. It is unknown whether
the effect of EDTA toxicity or the breaking of phytochela-
tin-metal bonds. However, EDTA does increase process
efficiency when soil experiments are conducted.
Additionally, hyperaccumulator status was achieved in
all studies for As and Cd. Ni hyperaccumulator status
was achieved in the presence three metals without EDTA.
Cr hyperaccumulator status was not reached in any of
the experiments performed.
Acknowledgements
The project was funded by the Ohio Plant Biotechnol-
ogy Consortium. The authors would like to thank Dr. Ran-
dall Mitchell, Biology Department at The University of
Akron. These experiments would not have been possible
without his generous use of his greenhouse space.
References
Bagga, D., Peterson, S., 2001. Phytoremediation of arsenic-contaminated
soil as affected by the chelating Agent CDTA and different levels of soil
pH. Remediation J. 12, 77–85.
Chen, H., Cutright, T., 2001. EDTA and HEDTA effects on Cd, Cr, and
Ni uptake by Helianthus annuus. Chemosphere 45, 21–28.
Cosio, C., Vollenweider, P., Keller, C., 2006. Localization effects of
cadmium in leaves of a cadmium-tolerant willow (Salix viminalis L.) I.
Macrolocalization and phytotoxic effects of cadmium. Environ. Exp.
Bot. 58, 64–74.
do Nacimento, C., Amarasiriwardena, D., Baoshan, X., 2006. Compar-
ison of natural organic acids and synthetic chelates at enhancing
phytoextraction of metals from a multi-metal contaminated soil.
Environ. Pollut. 140, 114–123.
Evangelou, M., Ebel, M., Scaeffer, A., 2006. Evaluation of the effect of
small organic acids on phytoextraction of Cu and Pb from soil with
tobacco Nicotiana tabacum. Chemosphere 63, 996–1004.
 M.C. January et al. / Chemosphere 70 (2008) 531–537
Gulz, P., Gupta, S., Schulin, R., 2005. Arsenic accumulation of common
plants from contaminated soil. Plant Soil 272, 337–347.
Gupta, A.K., Sinha, S., 2007. Phytoextraction capacity of the plants
growing on tannery sludge dumping sites. Bioresource Technol. 98,
1788–1794.
Hernandez-Allica, J., Garbisu, C., Barrutia, O., Becerril, J., 2007. EDTA-
induced heavy metal accumulation and phytotoxicity in cardoon
plants. Environ. Exp. Bot. 60, 26–32.
Kamnev, A., van der Lelie, D., 2000. Chemical and biological parameters
as tools to evaluate and improve heavy metal phytoremediation.
Biosci. Rep. 29, 239–258.
Keller, C., Hammer, D., 2004. Metal availability and soil toxicity after
repeated croppings of Thlaspi caerulescens in metal contaminated soils.
Environ. Pollut. 131, 243–254.
Li, Y., Dhankher, O., Carreira, L., Lee, D., Chen, A., Schroeder, J.,
Balish, R., Meagher, R., 2004. Over-expression of phytochelatin
synthase in Arabidopsis leads to enhanced arsenic tolerance and
cadmium hypersensitivity. Plant Cell Physiol. 45, 1787–1797.
Liphadzi, M.S., Kirkham, M.B., 2006. Availability and plant uptake of
heavy metals in EDTA-assisted phytoremediation of soil and com-
posted biosolids. S. Afr. J. Bot. 72, 391–397.
Luo, C., Shen, Z., Lou, L., Li, X., 2006. EDDS and EDTA-enhanced
phytoextraction of metals from artificially contaminated soils and
residual effects of chelant compounds. Environ. Pollut. 144, 862–871.
Madrid, F., Liphadzi, M., Kirkham, M.B., 2003. Heavy metal displace-
ment in chelate-irrigated soil during phytoremediation. J. Hydrol. 272,
107–119.
Marchiol, L., Assolari, S., Sacco, P., Zebri, G., 2004. Phytoextraction of
heavy metals by canola (Brassica napus) and radish (Raphanus sativus)
grown on multi-contaminated soil. Environ. Pollut. 132, 21–27.
Mari, S., Gendre, D., Pianelli, K., Ouerdane, L., Lobinski, R., Briat, J.F.,
Lebrun, M., Czernic, P., 2006. Root-to-shoot long-distance circulation
of nicotinamine and nicotinamine-nickel chelates in the metal hyper-
accumulator Thlaspi caerulescens. J. Exp. Bot. 57, 4111–4122.
Meers, E., Ruttens, A., Hopgood, M.J., Samson, D., Tack, F.M.G., 2005.
Comparison of EDTA and EDDS as potential soil amendments for
537
enhanced phytoextraction of heavy metals. Chemosphere 58, 1011–
1022.
Murillo, J., Maranon, T., Cabrera, F., Lopez, R., 1999. Accumulation of
heavy metals in sunflower and sorghum plants affected by Guadiamar
spill. Sci. Total Environ. 242, 281–292.
Ohio EPA (OEPA), 2005. 2003 Toxic Release Inventory, <www.
oepa.state.oh.us>.
Roosens, N., Verbruggen, N., Meerts, P., Ximenez-Embun, P., Smith, J.,
2003. Natural variation in cadmium tolerance and its relationship to
metal hyperaccumulation for seven populations of Thlaspi caerulescens
from western Europe. Plant Cell Environ. 10, 1657–1672.
Ruley, A.T., Sharma, N.C., Sahi, S.V., Singh, S., Sajwan, K., 2006. Effects
of lead and chelators on growth, photosynthetic activity, and Pb
uptake in Sesbania drumondii grown in soil. Environ. Pollut. 144, 11–
18.
Turgut, C., Pepe, M.K., Cutright, T.J., 2005. The effect of EDTA on
Helianthus annuus uptake, selectivity, and translocation of heavy
metals when grown in Ohio, New Mexico and Colombia soils.
Chemosphere 58, 1087–1095.
Wu, L., Luo, Y., Xing, X., Christie, P., 2004. EDTA-enhanced
phytoremediation of heavy metal contaminated soil with Indian
mustard and associated potential leaching risk. Agr. Ecosyst. Environ.
102, 307–318.
Wycisk, E., Kim, E., Schroeder, J., Kramer, U., 2004. Enhancing the first
enzymatic step in the histidine biosynthesis pathway increases free
histidine pool and nickel tolerance in Arabidopsis thaliana. FEBS Lett.
578, 128–134.
Yang, X., Feng, Y., He, Z., Stoffella, P., 2005. Review: molecular
mechanisms of heavy metal hyperaccumulation and phytoremediation.
J. Trace Elem. Med. Bio. 18, 339–353.
Zhang, P., Ye, Z., Lan, C., Xie, Z., Shu, W., 2005. Chemically assisted
phytoextraction of heavy metal contaminated soils using three plant
species. Plant Soil 276, 153–162.