REGULAR ARTICLE

Aspects of phytoremediation for chromium contaminated

sites using common plants Urtica dioica, Brassica napus
and Zea mays
Khaled Mahmud Shams & Gottfried Tichy &
Axel Fischer & Manfred Sager & Thomas Peer &
Ashtar Bashar & Kristina Filip
Received: 26 January 2009 / Accepted: 26 June 2009 / Published online: 9 July 2009
# Springer Science + Business Media B.V. 2009
Abstract As chromium is widely used in many
industries of which leather industries are the biggest
consumers, wastes from tanneries pose a serious
threat to the environment. Present research conducted
greenhouse pot experiments on several plants using
different solutions to find out their phytoextraction
potential. Among the plants, Urtica dioica found to
be very effective due to its higher uptake capacity for
chromium. The analyses showed that with 500 mg/
l solutions Cr concentration in roots was 38.9%
higher (average 20 mg/kg) than plants fed with
200 mg/l solutions (phytostabilisation), while with
200 mg/l solutions more Cr was transported to leaves
(phytoextraction), 10.52% higher (average 9.5 mg/kg)
concentration than plants fed with 500 mg/l solutions,
but also means that plants were less affected by
toxicity. Zea mays showed high tolerance towards Cr
with negligible concentration in leaves. Although
average Cr concentration in Brassica napus is lower
(25% and 21% for 200 mg/l and 500 mg/l solutions
respectively) than Urtica dioica which still could
account for phytoextraction, but due to its high
vulnerability to insects its potentiality has down-
graded. Among other elements, K being a component
of the salt, played significant role for plants growth
and survivalopening new avenue for phytoreme-
diation of contaminated sites.
Keywords Phytoremediation
.
Phytoextraction
.
Phytostabilization
.
Urtica dioica
.
Brassica napus
.
Zea mays
Introduction
Rapid industrialization, especially in developing
countries like India, China, Brazil, and Bangladesh, has
led to serious environmental pollution and has become a
threat to the local ecosystems. The possibility to clean
the contaminated environment with plant biota is
drawing much attention from environmental scientists
against the traditional expensive site clean-up technolo-
Plant Soil (2010) 328:175189
DOI 10.1007/s11104-009-0095-x
Responsible Editor: Juan Barcelo.
K. M. Shams (*)
:
G. Tichy
Department of Geography and Geology,
University of Salzburg,
Salzburg, Austria
e-mail: khaled.shams@geo.uni-halle.de
A. Fischer
:
K. Filip
Clausthaler Umwelttechnik-Institute GmbH,
Clausthal-Zellerfeld, Germany
M. Sager
Austrian Agency for Health and Food Safety,
Vienna, Austria
T. Peer
Department of Organism Biology, University of Salzburg,
Salzburg, Austria
A. Bashar
Institute for Geotechnical Engineering and Mine Surveying,
TU Clausthal, Germany
gies. Classic methods, such as excavation, thermal
treatment and chemical soil washing are typically
expensive and destructive (Wenzel et al. 2004).
Phytoremediation, use of proper plants to clean
contaminated sites (USEPA 2001; Shann 1995;
Chaney et al. 1997; Dietz and Schnoor 2001; Suresh
and Ravishankar 2004; Sing and Jain 2003; Odjegba
and Fasidi 2007), is an emerging technology that
employs the use of plants for clearing the contami-
nated areas. Plant cultivation and harvesting are
inexpensive processes compared with traditional
engineering approaches involving intense soil manip-
ulation (Singh and Ma 2003). Fundamental and
applied research have unequivocally demonstrated
that selected plant species possess the genetic poten-
tial to remove, degrade, metabolize or immobilize a
wide range of contaminants (Lasat 2000).
Due to technological advancements, metal releases
by industrial activities have been strongly reduced, as
a consequence, metal concentrations in the air have
been significantly decreased. However, accumulating
metals in soils are still leading to higher risks due to
leaching into ground and surface water, uptake by
plants and direct or indirect intake by human
population. When present at increased levels of
bioavailability, both essential (Cu, Zn, Mn, Fe, Ni,
Mo) and non-essential metals (Cd, Pb, Hg, Cr) are
toxic (Krenlampi et al. 2000).
Main objectives of the present study are:
1. To focus on phytoextraction potential of selected
green plants for Cr from Cr-contaminated soils.
As Cr is widely used industrial chemicals,
therefore plants grown universally were selected
for the experiment.
2. To appraise the role of K for plants growth and
survival as it is an inherent constituent of the
chromium salts used in experiment and simulta-
neously present in garden mould, and to compare
plants behaviour towards other nutrients (N, P, and
S) present in garden mould and in chromium salt.
Material and methods
Plants used in the experiments
Urtica dioica L. (subspecies dioica), commonly
known as Stinging nettle (in German: Brennnessel)
belongs to the family of Urticaceae. The leaves of the
plants have stinging hairs, causing irritation to the
skin. The plant grows in the temperate regions
throughout the world. Natural habitat includes waste
ground, hedgerows, woods etc. with perennial growing
to 1.2 m by 1 m at a fast rate (Fern 1997). Several
studies have been conducted on heavy metals (Cu,
Zn, Pb, Cd, and Hg) accumulation in Urtica dioica
(Khan and Joergensen 2006; Notten et al. 2005;
Edwards et al. 2004), but there is scarcity of data on
Cr uptake by Urtica dioica. As this plant showed
great potential for other heavy metals accumulation,
and as it is available in tropical climate, therefore
present study emphasized on Urtica dioica as a
potential plant for Cr-contaminated sites.
Brassica napus L. commonly known as Rapeseed
is a bright yellow flowering member of the family
Brassicaceae (Rapeseed or cabbage family). Experi-
ments by Zayed et al. (1998) on different vegetable
plants including variety of Brassicaceae (Cauliflower,
Cabbage, broccoli etc.) found root to shoot transloca-
tion of Cr extremely limited and root accumulation
100-fold higher regardless of the Cr species supplied. It
has been reported that Indian mustard (Brassica
juncea) accumulates significant amount of Cr in both
shoots and roots at higher soil-Cr concentrations
despite severe phytotoxic symptoms (Han et al.
2004). Experiments at the present study were con-
ducted on rapeseed to investigate its response on Cr
and thereafter its uptake capacity of the toxic metal.
Maize (Zea mays L.) also known as corn is a
popular cereal grain in many countries worldwide.
Hybrid maize can grow several meters while it is about
1 m in height in most cases. Maize is increasingly used
as a biomass fuel, such as ethanol, which as researchers
search for innovative ways to reduce fuel costs has
unintendly caused a rapid rise in food costs recently.
Other than as food or biofuel, several investigations
have been carried out on maize to determine its
potential as contaminant accumulator in polluted sites
for Cd and As (Lee et al. 1998; Yi et al. 2003). Due to
its versatile nature and unexplored potentiality to many
other toxic metals, maize is included to the pot
experiments to investigate its sensitivity to Cr.
Experimental approach
The phytoremediation experiments were conducted in
summer 2006, from July 20 to October 9 in the
176 Plant Soil (2010) 328:175189
greenhouse of the University of Salzburg. After two
and half months, plants were brought to the laboratory
for analyses. The greenhouse experiments were con-
ducted on garden mould or Blumenerde and obtained
from local shops. It is worth to mention here that of the
three plants only Zea mays blossomed. Brassica napus
fed with 50 mg/l solutions could not survive till the end
of experiment. As the survival rate and growth of
Urtica dioica fed with 50 mg/l solutions were low,
therefore, they were not analyzed considering the low
nutrient impacts of K in solutions being the main factor
for both.
Typical characteristics of the garden mould are as
follows: pH (CaCl2) between 5.5 to 6.5, salt content
in g KCl/l 1.02.0. Other nutrients include 50
300 mg/l N, 40 to 300 mg/l P2O5 and 100500 mg/
l K2O.
1 liter plastic pots were used for Urtica dioica and
Brassica napus plants while 0.8 liter pots were used
for Zea mays as the number of plants per pot is half of
the former two. Each pot contained four plants in case
of Urtica dioica and Brassica napus, and 2 plants of
Zea mays. Seeds were obtained from the Botanical
Garden of the University of Salzburg and nurtured
carefully to grow the seedlings. Healthy seedlings
were then put into pots filled with garden mould, and
treated with chemical grade chromium potassium
sulfate (KCr(SO4)2.12H2O, Merck p.a.) as Cr3+
source. The minimum solubility of the chemical is
99% and molecular weight 499.41 g/mol.
Chromium treatments as Cr-solution included
control (0), 50, 200, and 500 mg/l solutions. Each
treatment had five replicas. All pots were watered
regularly (about 100 ml per pot) in the morning and
were allowed to drain to prevent accumulation of salts
by evapotranspiration. Watering was done manually.
The plants were treated with Cr-solutions in every
alternate day.
After harvesting, plant samples were air dried for
2 days and finally dried in the Oven at 80C for 24 h.
The dried samples were separated into three portions,
leaves, stems and roots, and crashed into fine powder
for analysis. Roots were mildly shaken to leave off
the loose soil from its body but only to let the
rhizospheric soil particles that were closely attached
to it to remain with the roots for analysis.
For elemental analysis (Cr, K, P and S) total
digestion of dried plant samples was done in PTFE
pressure bombs with microwave assisted heating in an
mls 1200 mega high performance microwave digestion
unit. About 0.3 g dried sample was mixed with 3.5 ml
65% HNO3 suprapure and 1 ml 30% H2O2 suprapure
in the digestion vessel, closed, and submitted to a
maximum of 600 Watts. After cooling, the sample
solutions were transferred to a capped plastic vessel, and
made up to 10 g on a balance. Main and trace elements
were determined by ICP-OES (Inductively Coupled
Plasma Optical emission Spectrometry) on a Perkin-
Elmer 3000 XL fitted with an axial torch.
N digestion and distillation were carried out using
a Bchi 321 digestion unit (Bchi Laboratoriums-
Technik AG, Switzerland). Sample weights ranged
from 81.81 to 181.85 mg. Samples were weighed in
an analytical balance, and placed in a digestion flask.
3.5 g of catalyst and 20 ml concentrated H2SO4 were
added to each flask. Digestion was carried out until
the solution was completely clear. Digestion time was
between 90 and 120 min. Once digestion was
complete, the sample was distilled in a distillation
flask (glass trap) and a condenser into a highly
alkaline solution by addition of 90 ml of 32% (w/v)
NaOH. Released ammonium was caught in 30 ml
boric acid. The titer of the ammonium borate formed
was measured by addition of 0.02 N hydrochloric
acid, to a pH value<4.8.
Results presented here are from the data available
from analyses. Data for root concentration of Cr in
Brassica napus and stem concentration of Cr in Zea
mays were not available for interpretation.
Results
Plant growth
Leaves of Urtica dioica plants fed with 500 mg/
l chromium solutions were not as green as with
200 mg/l solutions. Many leaves turned to be
yellowish, yellow spots were observed. All four
plants in each pot survived well although a variation
in growth and size was remarkable. Plants survival
rate with 200 mg/l solutions was 90%. Poor growth
was observed with 50 mg/l solution with a 60%
survival rate while plants in control pots showed
extremely poor growth.
Brassica napus plants with 500 mg/l chromium
solution showed better growth than plants with
200 mg/l. While plants with 500 mg/l solution
Plant Soil (2010) 328:175189 177
survived 100%, with 200 mg/l solutions only 65%
plants survived. Plants in the control pots and with
50 mg/l solutions could not survive the time span of
the experiment.
Zea mays grew well with all solution concentra-
tions (100% survival rate) but showed better growth
with higher concentrations while plants growth in
control pots were very poor. Figures 1 and 2 show the
maximum growth, height and roots in relation to
different Cr-solutions for Urtica dioica, Brassica
napus and Zea mays.
Figures 3a, b, and c show plant weight for Urtica
dioica, Brassica napus and Zea mays. The discrep-
ancies can be attributed to the small size of the pots
which are very limited to contain and supply nutrients
equally to each plant. Competition of plants among
each other for nutrients is another determining factor
that let some plants grew very well while others
lacked sufficient nutrition. While one plant of Urtica
dioica produced 10.6 g in fresh weight other in the
same pot produced only 1.6 g in pots fed with
500 mg/l solution. The 85% higher growth clearly
exhibits a competition of plants among themselves for
nutrients. Maize showed an increasing trend of better
growth with increasing concentration of Cr-solutions
in the order Zea mays (500 mg/l) > (Zea mays
200 mg/l) > Zea mays (50 mg/l).
For Urtica dioica mean total dry weight was
from 15% to 22% of total weight while for Brassica
napus it was 1819% of total weight. Zea mays had
dry weights ranging from 24% to 43% of its total
weight.
Special situation in the greenhouse
The greenhouse pot experiments, especially on
Brassica napus and lesser on Urtica dioica experi-
enced some harmful infestation from insects. A large
number of tiny flies were abundant around Urtica
dioica and managed manually to keep the plants
Urtica dioica Brassica napus
Zea mays
0
10
20
30
40
50
50 mg/l 200 mg/l 500 mg/l
Solution concentrations
R
o
o
t

l
e
n
g
t
h

(
c
m
)
Fig. 2 Root lengths
(cm) with different
concentrations of
Cr-solutions for Urtica
dioica, Brassica napus
and Zea mays. Brassica
napus plants with 50
mg/l solution could not
survive the experiment
Urtica dioica
Brassica napus
Zea mays
0
50
100
150
50 mg/l 200 mg/l 500 mg/l
Solution concentrations
H
e
i
g
h
t

(
c
m
)
Fig. 1 Plant heights
(cm) with different
concentrations of
Cr-solutions for Urtica
dioica, Brassica napus
and Zea mays. Brassica
napus plants with 50
mg/l solution could not
survive the experiment
178 Plant Soil (2010) 328:175189
growing until all insects could be eradicated. Brassica
napus on other hand were severely affected by the
insects which ate the young leaves and were also
controlled manually until the last surviving insects
found. Zea mays, on the other hand, found to be
resistant against insects and were only affected by
nutrition added to it accordingly.
Cr, K, P, S, and N contents in plants
Urtica dioica (Stinging nettle)
Average Cr concentration in leaves of Urtica dioica
fed with 200 mg/l solutions ranges from 5 to 15 mg/kg
while plants fed with 500 mg/l solutions showed a rather
0
5
10
A B C D E
A B C D E
A B C D E F
Urtica dioica
P
l
a
n
t

w
e
i
g
h
t

(
g
)
P
l
a
n
t

w
e
i
g
h
t

(
g
)
P
l
a
n
t

w
e
i
g
h
t

(
g
)
Legend
200 mg/l
500 mg/l
0
10
20
30
40
Brassica napus
Legend
0
20
40
60
80
Zea mays
50 mg/l
200 mg/l
500 mg/l
Legend
200 mg/l
500 mg/l
b
c
Fig. 3 a Mean plant weight
for Urtica dioica with
reference to 200 mg/l and
500 mg/l solutions in Pots A
to E. Error bars indicate
standard deviation. b Mean
plant weight for Brassica
napus with reference to
200 mg/l and 500 mg/l
solutions in Pots A to E.
Error bars indicate standard
deviation. c Mean plant
weight for Zea mays with
reference to 50 mg/l,
200 mg/l, and 500 mg/l
solutions in Pots A to
F. Error bars indicate
standard deviation
Plant Soil (2010) 328:175189 179
homogenized concentration at 10 mg/kg (Fig. 4). Data
showed plants with 500 mg/l solutions consumed more
Cr in its leaves than the plants fed with 200
mg/l solutions. Cr concentrations in stems were 1 mg/
kg for both groups suggesting nettles are less capable
of storing Cr in stems than in leaves (Fig. 5). In
general, Cr concentrations in nettle leaves were nine
times higher than stems. Root concentration of Cr
varied accordingly with solution concentrations in both
groups. For plants fed with 200 mg/l solutions, Cr
concentration was 12 mg/kg while for plants fed with
500 mg/l it was 20 mg/kg (Fig. 5). Figure 6 shows the
shoot/root ratio of Cr for both groups which was 82%
higher in case for plants fed with 200 mg/l solution
(0.867) than the group fed with 500 mg/l solution
(0.475).
While average K concentration in leaves of Urtica
dioica decreased from 13098 mg/kg (200 mg/l solution)
to 10820 mg/kg (500 mg/l solutions), it increased as
high as 42% from 973 to 1383 mg/kg in roots
suggesting plants accumulated more K in roots with
the availability of this nutrient although the quantity is
remarkably higher in leaves. This also points to the
priority of plants to survive at the first place as toxicity
rises, then to grow and act as remediation medium. P
concentration in roots increased from 225 mg/kg
(200 mg/l solution) to 304 mg/kg (500 mg/l solution,
26% increase) but in leaves it decreased from 3791 mg/
kg to 3271 mg/kg for 200 and 500 mg/l solutions
respectively. Increase of both K and P concentration in
roots could be the main reason of survival for all four
plants of Urtica dioica with 500 mg/l solutions. Figure 7
shows the shoot to root ratio of K and P in Urtica
dioica. As the ratio decreased, more K and P were
accumulated in roots with higher solution concentra-
tions. It is to be mentioned here that K and P are the
primary nutrients for plants growth.
0
5
10
15
200 mg/l 500 mg/l
Urtica dioica
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
Fig. 4 Mean leave concentration of Cr in Urtica dioica fed
with 200 mg/l and 500 mg/l solutions respectively. Error bars
indicate standard deviation
0,0
0,5
1,0
200 mg/l 500 mg/l
Urtica dioica
S
h
o
o
t
/
R
o
o
t
Fig. 6 Shoot to root ratio of Cr in Urtica dioica with 200 mg/l and
500 mg/l solutions
Legend
0
5
10
15
20
25
200 mg/l 500 mg/l
Urtica dioica
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)Root
Stem
Fig. 5 Total Cr concentra-
tions in roots and stems of
Urtica dioica with 200 mg/
l and 500 mg/l solutions
180 Plant Soil (2010) 328:175189
Sulfur concentration in both shoot and root was
higher with higher solution concentrations. Shoot
concentration increased from 1536 mg/kg (200
mg/l solutions) to 1715 mg/kg (with 500 mg/l sol-
utions). In roots S concentration increased up to 27%
from 97 mg/kg to 132 mg/kg for 200 and 500
mg/l solutions. As water-extractable S is very low in
the soil, mean: 4.8 mg/l (data not shown here), therefore
major source of S in plants is the chemicals used to feed
the plants (KCr(SO4)2.12H2O) (Fig. 8).
N concentration in both shoot and root of Urtica
dioica with 200 mg/l solutions was higher than the
plants with 500 mg/l solutions. In leaves N concen-
tration was 1317 mg/kg and 1096 mg/kg respectively
while in root it was 227 and 137 mg/kg respectively
(Fig. 9). As sulfur is an important constituent of
proteins, its elevated concentration in plants with
500 mg/l solution played a complementary role for N
causing a lowering of N level in plants with 500
mg/l solutions. It is to be noted that as same quantity of
garden mould is used for all pots, therefore N content
remained constant throughout the experiment.
Brassica napus (Rapeseed)
Figure 10 shows Cr concentrations in leaves and
stems in Brassica napus. Leave concentrations of Cr
in plants fed with 200 mg/l solutions were higher than
plants fed with 500 mg/l solutions. Average Cr
concentrations in stems were almost equal to leaves.
In contrast to Urtica dioica, rapeseed did not show
any significant differences for Cr concentrations in
leaves and stems.
Rapeseed plants required more K and P for its
growth than nettles (Figs. 11 and 12). Leave concen-
trations of K were 2.1 and 1.5 times higher with 200
and 500 mg/l solutions respectively than the
corresponding nettles. Stem concentrations were 7.1
and 4.1 times higher for both solutions than the
nettles. Concentration of P was higher in Brassica
napus than in Urtica dioica as well. While with
500 mg/l solutions leave concentrations were almost
same as for Urtica dioica but with 200 mg/l solution
P concentration was 1.8 times higher.
As data shows (Fig. 13), due to higher concentra-
tion of S in solution, Brassica napus fed with 500
mg/l solutions consumed slightly higher S in leaves
and stems than the plants fed with 200 mg/l solutions.
Moreover concentrations in stems were 8.8 and 5.2
times higher than Urtica dioica fed with 200 and
500 mg/l solutions respectively, while leaf concen-
trations in both the species were almost equal (1.1
Legend
0
5
10
15
20
200 mg/l 500 mg/l
Urtica dioica
S
h
o
o
t
/
R
o
o
t
K
P
Fig. 7 Shoot to root ratio of K and P in Urtica dioica with
200 mg/l and 500 mg/l solutions
Legend
0
500
1000
1500
2000
2500
3000
200 mg/l 500 mg/l
Urtica dioica
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
Leave
Stem
Root
Fig. 8 Mean S concentra-
tions in leaves of Urtica
dioica with 200 and
500 mg/l solutions. Error
bars indicate standard
deviation. Stems and
roots correspond to total
concentration
Plant Soil (2010) 328:175189 181
times higher in Urtica dioica with both solution
concentrations). Similarly, N concentration in rape-
seed leaves and stems were significantly higher than
nettles (Fig. 14). Leave concentrations were 10 and 7
times higher than Urtica dioica with 200 and 500 mg/
l solutions respectively. Stem concentrations were 65
and 49 times higher than the corresponding Urtica
dioica. Above results indicate higher N and S
requirements by Brassica napus for its growth and
survival.
Zea mays (Maize)
Cr concentrations in leaves of maize plants were very
low compared to Urtica dioica and Brassica napus.
Average Cr concentrations in leaves were 0.24 mg/kg,
0.34 mg/kg and 0.38 mg/kg for plants fed with
50 mg/l, 200 mg/l and 500 mg/l solutions respectively
(Fig. 15a), while the average root concentrations were
significantly higher being 8 mg/kg, 12 mg/kg and
14 mg/kg respectively (Fig. 15b). As the roots of Zea
mays in experimental pots could grow more than
30 cm, therefore it can aid via phytostabilization for
deeper subsurface pollutions.
Figures 16 and 17 show K and P concentrations in
leaves and roots of Zea mays. K concentration in
leaves is 1.1 and 1.3 times higher than nettles,
whereas P in Urtica dioica leaves is 1.6 and 2.2
times higher for both 200 and 500 mg/l cultures
respectively. Root concentration of K was 3 and 2 times
higher than nettles for 200 and 500 mg/l cultures
respectively. P concentrations were 4.5 and 3 times
higher than in Urtica dioica. Figure 18 shows the ratio
of K and P in leaves vs. root in maize. It clearly shows
Legend
0
500
1000
1500
2000
200 mg/l 500 mg/l
Urtica dioica
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
Leave
Stem
Root
Fig. 9 Mean N concentra-
tions in leaves of Urtica
dioica with 200 and
500 mg/l solutions. Error
bars indicate standard
deviation. Stems and
roots correspond to total
concentration
Legend
0
1
2
3
4
5
200 mg/l 500 mg/l
Brassica napus
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
Leave
Stem
Fig. 10 Mean Cr
concentrations in leaves
and stems of Brassica napus
with 200 mg/l and 500
mg/l solutions. Error bars
indicate standard deviation
182 Plant Soil (2010) 328:175189
plants increased affinity for K with increasing solution
concentrations while P concentrations decreased
accordingly.
Plant dry weight and Cr content
Figure 19a and b show a relation of plant dry weight
vs. Cr concentrations in leaves. Whereas nettle
produced very little dry weight with 200 mg/l solutions
(mean: 0.75 g) but it concentrated significantly higher
Cr in leaves than others (mean: 9.5 mg/kg). Rapeseed
showed moderate data of dry weight vs. Cr concentra-
tion which was 3.26 gm vs. 2.4 mg/kg chromium.
Maize on the other hand produced about 10 gm in dry
weight but its concentration of Cr in leaves was
negligible (about 0.34 mg/kg).
With 500 mg/l solutions nettles produced on
average 0.61 gm in dry mass, and rapeseed produced
2.63 gm but concentrations of Cr in leaves was
significantly higher in nettle leaves (8.5 mg/kg) than
in rapeseed (1.8 mg/kg). Maize showed a better
growth producing 15 gm in dry mass with a very
minimal Cr concentration in leaves (0.38 mg/kg).
In general, mean dry weight (higher to lower): Zea
mays > Brassica napus > Urtica dioica, and mean Cr
concentrations in leaves (higher to lower) Urtica
dioica> Brassica napus > Zea mays.
The dry weight vs. Cr concentrations suggests
Urtica dioica as the best plant for Cr contaminated
sites. While Zea mays showed higher tolerance against
Cr and achieved its complete growth despite high
concentration of Cr in solutions but its not an effective
plant for phytoextraction. Brassica napus are highly
vulnerable to insects and because of extra attention it
needs to optimize its full potential, it can be of least
choice for sites contaminated with chromium.
Legend
0
10000
20000
30000
40000
50000
60000
200 mg/l 500 mg/l
Brassica napus
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)Leave
Stem
Fig. 11 Mean K concentra-
tions in leaves and stems
of Brassica napus with
200 mg/l and 500 mg/l
solutions. Error bars
indicate standard deviation
Legend
0
2000
4000
6000
8000
10000
200 mg/l 500 mg/l
Brassica napus
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
Leave
Stem
Fig. 12 Mean P concentra-
tions in leaves and stems
of Brassica napus with
200 mg/l and 500 mg/l
solutions. Error bars
indicate standard deviation
Plant Soil (2010) 328:175189 183
Soil-plant final Cr accumulation
Figure 20 shows final Cr concentration in pot soils for
Urtica dioica, Brassica napus, and Zea mays. Meas-
urements were done on a randomly chosen single pot
only for each solution concentration for each group of
plants. Soils used for Urtica dioica and Brassica napus
showed quite close concentration, but for Zea mays
concentration differed markedly. Soils for nettles with
200 mg/l solution reached the final Cr concentration of
252 mg/kg and for 500 mg/l it was about 468 mg/kg.
Soils for rapeseed had Cr concentrations of 231 mg/kg
and 537 mg/kg for 200 and 500 mg/l solutions
respectively. Soils for maize plants had a concentration
of 22 mg/kg, 90 mg/kg and 212 mg/kg for 50, 200 and
500 mg/l solutions respectively. As data for stem
concentrations for maize were not available, therefore,
it could not be concluded as a reason that maize
accumulated substantial amount of Cr in its stem.
Figure 21 shows the ratio of final soil vs. average
plant above-ground concentrations of Cr for nettle,
rapeseed and maize. Although the graph shows none
of the plants as hyperaccumulator but the above-
ground concentrations of Cr in nettles are remarkable
(about 10 mg/kg for both cultures). This is promising
as Urtica dioica exhibited a unique stable response
towards Cr uptake in its leaves. It is worth to mention
here that the experiments were carried out without
any chelating agents that could artificially enhance its
uptake capacity, and as Cr is supplied in trivalent
form which in the pH range of 5.5 to 6.5 precipitates,
thus becoming more immobile and difficult for plants
to absorb. Therefore, Urtica dioica with a mild
presence of K in the chemicals produced very
Legend
0
500
1000
1500
2000
200 mg/l 500 mg/l
Brassica napus
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
Leave
Stem
Fig. 13 Mean S concentra-
tions in leaves and stems
of Brassica napus with
200 mg/l and 500 mg/l
solutions. Error bars
indicate standard deviation
Legend
0
5000
10000
15000
20000
25000
200 mg/l 500 mg/l
Brassica napus
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
Leave
Stem
Fig. 14 Mean N concentra-
tions in leaves and stems
of Brassica napus with
200 mg/l and 500 mg/l
solutions. Error bars
indicate Standard deviation
184 Plant Soil (2010) 328:175189
promising results to be considered as a unique plant
for chromium remedial purposes.
Discussion
As nettle (rapeseed could not survive the experimental
time span) did only produce negligible amounts of
biomass at 50 mg/l solution concentration, it clearly
indicates with comparison to 200 mg/l and 500 mg/l,
that K had been the dominating plant survival and
growth factor throughout the experiment. Although
Urtica dioica is more resistive to grow in adverse
environmental conditions but Cr could already hinder
plant growth from 50 mg/l solution concentration. At
higher concentrations plants showed increased uptake
0,0
0,5
1,0
50 mg/l 200 mg/l 500 mg/l
Zea mays
50 mg/l 200 mg/l 500 mg/l
Zea mays
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
0
10
20
30
a
b
Fig. 15 a Mean Cr concen-
trations in leaves of Zea
mays with 50 mg/l, 200
mg/l and 500 mg/l solutions.
Error bars indicate standard
deviation. b Mean Cr
concentrations in roots of
Zea mays with 50 mg/l,
200 mg/l and 500 mg/l
solutions. Error bars
indicate standard deviation
Legend
0
10000
20000
30000
40000
50000
50 mg/l 200 mg/l 500 mg/l
Zea mays
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
Leave
Root
Fig. 16 Mean K concentra-
tions in leaves and roots
of Zea mays with 50 mg/l,
200 mg/l and 500 mg/l
solutions. Error bars
indicate standard deviation
Plant Soil (2010) 328:175189 185
of Cr and K. Hence addition of K played a very
significant role for the efficiency of nettles and
rapeseed as accumulator of toxic metal like Cr. Maize
on the other hand survived in all three phases but plant
growth was better with higher concentrations as well.
However, chromium concentration in leaves was 22 to
28 times lower than nettles and 5 to 7 times lower than
rapeseed, suggesting higher tolerance for Cr.
It is known that some plants can survive quite well
under elevated metal conditions without the hyper-
accumulation characteristic. These plants appear to
tolerate metals in their environment by using a variety
of mechanisms, including exudation of compounds
that complex metals in the contaminated environment,
thereby preventing their entry into the root in the first
place (Dzantor 2000). However, aim of the present
study was to find a hyperaccumulator plant rather
than a Cr-tolerant plant.
Root concentrations of Cr in maize plants were
close to nettles. Both plants can be considered
positive for phytostabilization of Cr contaminated
sites. However, maize is an edible plant and can easily
be grazed by cattles while stinging nettles because of
its unpleasant nature will distract animals away.
Therefore Zea mays does not conform with the aims
and objectives of the research.
Brassica napus or rapeseed regardless of its
substantial amount of chromium uptake in leaves
and stems does not stand as robust candidate for
remediation of Cr contaminated sites. Rapeseed is an
edible plant, and will be grazed by cattle or used by
humans as vegetables, thereby limiting its possibility
to be used in fields. Further on, it is vulnerable to
insects and needs pesticides to grow its full phase to
accomplish remedial purposes. Use of pesticides is
not desirable as it further threatens the eco-system in
general.
Higher Cr levels in soils has very well-documented
negative impact on plant root growth. Studies on Salix
viminalis (Prasad et al. 2001), Caesalpinia pulcherrima
Legend
0
2000
4000
6000
8000
10000
12000
50 mg/l 200 mg/l 500 mg/l
Zea mays
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
Leave
Root
Fig. 17 Mean P concentra-
tions in leaves and roots of
Zea mays with 50 mg/l,
200 mg/l and 500 mg/l sol-
utions. Error bars indicate
standard deviation
Legend
0,0
2,0
4,0
6,0
50 mg/l 200 mg/l 500 mg/l
Zea mays
S
h
o
o
t
/
R
o
o
t
K
P
Fig. 18 Shoot to root ratio
of K and P in Zea maize
with 50 mg/l, 200 mg/l, and
500 mg/l solutions
186 Plant Soil (2010) 328:175189
(Iqbal et al. 2001) and on mung bean (Samantaray
et al. 1999) showed significant root length decrease
with higher Cr concentrations in soil. Scanning
electron microscope studies of roots affected by Cr
showed increased growth of root hairs and increased
relative proportion of pith and cortical tissue layers
(Suseela et al. 2002). Under high concentrations of
both the Cr species combination, the reduction in root
growth could be due to the direct contact of seedlings
roots with Cr in the medium causing a collapse and
Legend
0
100
200
300
400
500
600
50 mg/l 200 mg/l 500 mg/l
Solution Concentrations
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)Urtica dioica
Brassica napus
Zea mays
Fig. 20 Cr concentration in pot soils with 50 mg/l, 200
mg/l and 500 mg/l solutions
Legend
0
0,01
0,02
0,03
0,04
0,05
50 mg/l 200 mg/l 500 mg/l
Solution concentrations
P
l
a
n
t
/
S
o
i
l
Urtica dioica
Brassica napus
Zea mays
Fig. 21 Cr concentration in soil vs. in above ground parts of
Urtica dioica, Brassica napus, and Zea mays. Data for Zea
mays shows Cr concentration in leaves only
Legend
0
4
8
12
Urtica dioica Brassica napus Zea mays
Urtica dioica Brassica napus Zea mays
W
e
i
g
h
t

(
g
)
W
e
i
g
h
t

(
g
)
0
2
4
6
8
10
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
C
o
n
c
e
n
t
r
a
t
i
o
n

(
m
g
/
k
g
)
wt. in gm
Cr in mg/kg
Legend
wt. in gm
Cr in mg/kg
0
4
8
12
16
0
2
4
6
8
10
a
b
Fig. 19 Dry weights vs. Cr
concentration in Urtica dio-
ica, Brassica napus and Zea
mays with 200 mg/l (a) and
500 mg/l (b) solutions
Plant Soil (2010) 328:175189 187
subsequent inability of the roots to absorb water from
the medium (Barcelo et al. 1986).
Although higher Cr concentration decreases root
length of plants as mentioned above, present research
showed an increase of root length and better plant
growth with higher chromium concentrations (Figs. 1
and 2). While plants are supposed to suffer from their
growth and survival because of toxicity effect of Cr, but
due to the chemistry of the solution where K is an
ingredient (KCr(SO4)2.12H2O) of the salt, plants
showed an unexpected positive response towards their
growth and uptake of chromium. This is very significant
as it can open a new avenue for environmentalists and
bioremediation experts to enhance Cr uptake naturally
without addition of any artificial chelating agents like
EDTA (ethylene diamine tetraacetic acid). As the salt
contained K, a key nutrient to plants, therefore even with
increasing solution concentrations (toxicity) plants grew
and survived the hazard.
The phytoextraction potential of Urtica dioica is
higher than Brassica napus and Zea mays. As data
showed Urtica dioica with 200 mg/l chromium
solution most promising (and as the differences with
500 mg/l were very thin), this finding is important as
it suggests that a mild application of potassium in
field can cause a greater impact for plants for its
growth and uptake of toxic metals like chromium
from contaminated sites. Thus potassium can be a
potential aid element in Cr-contaminated sites for
phytoremediation technology. The production of least
harvestable biomass (less than a gm) adds an
additional aspect characterizing Urtica dioica as the
only choice above Brassica napus and Zea mays.
Therefore, due to its higher Cr uptake and low
biomass production Urtica dioica, commonly known
as Stinging nettle, in German Brennnessel, can be
considered as the right plant for remediation of Cr-
contaminated sites. As nettle grows both in tropical and
cold climates, therefore its value as Nature Cleaner is
universal. Further experiments and analyses will reveal
more potential characteristics and usages of nettles
concerning environmental pollution. And it is highly
recommended.
Conclusion
As the experiment, conducted without addition of any
chelating agents, proved to be very effective with a
minimal application of K, therefore, it can be
hypothesized that Urtica dioica as a phytoremediation
plant with a minimal K of 200 mg/l applied in the
solution can remove substantial amount of Cr from
contaminated sites.
Acknowledgement Author is grateful to the Afro-Asiatische
Institute, Salzburg, Austria, for the financial support to carry
out this research project. Thanks are also due to Prof. Dr. Paul
Heiselmayer (Head of the Botanical Garden) for his permission
to carry the pot experiments in the greenhouse of the University
of Salzburg. Special thanks go to Elisabeth Egger (staff
member of the Botanical Garden) for her help and guidance
to grow and caring the plants till the end of the experiments.
References
Barcelo J, Poschenrieder C, Gunse B (1986) Water Relations of
Chromium VI Treated Bush Bean Plants (Phaseolus
vulgaris L cv Contender) Under Both Normal and Water
Stress Conditions. J Exp Bot 37:17887
Chaney RL, Malik M, Li YM, Brown SL, Angle JS, Baker AJM
(1997) Phytoremediation of soil metals. Curr Opin Biotechnol
8:279284
Dietz AC, Schnoor JL (2001) Advances in Phytoremediation.
Environ Health Perspect 109(suppl 1):163168
Dzantor EK (2000) Phytoremediationplant-based strategies for
cleaning up contaminated soils: metal contaminants. Fact
sheet 768. Maryland Cooperative Extension, University of
Maryland.
Edwards SC, MacLeod CL, Lester JN (2004) The bioavailability
of copper and mercury to the common nettle (Urtica dioica)
and the earthworm eisenia fetida from contaminated dredge
spoil. Water Air Soil Pollut 102:7590
Fern K (1997) Plants for a futureedible & useful plants for a
healthier world. Permanent Publications, Hyden House Ltd
Han FX, Sridhar BBM, Monts DL, Su Y (2004) Phytoavailability
and toxicity of trivalent and hexavalent chromiumto Brassica
juncea. New Phytol 162:489499
Iqbal MZ, Saeeda S, Shafiq M (2001) Effects of Chromium on
an important arid tree (Caesalpinia pulcherrima) of
Karachi city. Pakistan Ekol Bratislava 20:41422
Krenlampi S, Schat H, Vangronsveld J, Verkleij JAC, van der
Lelie D, Mergeay M, Tervahauta AI (2000) Genetic
Engineering in the Improvement of Plants for Phytoremedia-
tion of Metal Polluted Soils. Environ Pollut 107:225231
Khan KS, Joergensen RG (2006) Decomposition of heavy
metal contaminated nettles (Urtica dioica L.) in soils
subjected to heavy metal pollution by river sediments.
Chemosphere 65(6):981987
Lasat MM (2000) Phytoextraction of metals from contaminated
soil: a review of plant/soil/metal interaction and assessment
of pertinent agronomic issues. J Hazard Subst Res 2:125
Lee J-S, Chon H-T, Kim K-W (1998) Migration and dispersion
of trace elements in the rock-soil-plant system in areas
underlain by black shales and slates of the Okchon Zone,
Korea. J Geochem Explor 65(1):6178
188 Plant Soil (2010) 328:175189
Notten MJM, Oosthoek AJP, Rozema J, Aerts R (2005) Heavy
metal concentrations in a soil-plant-snail food chain along
a terrestrial soil pollution gradient. Environ Pollut 138
(1):178190
Odjegba VJ, Fasidi IO (2007) Phytoremediation of heavy metals
by Eichhornia crassipes. Environmentalist 27:349355
Prasad MNV, Greger M, Landberg T (2001) Acacia nilotica L
bark removes toxic elements from solution: corroboration
from toxicity bioassay using Salix viminalis L in hydroponic
system. Int J Phytoremed 3:289300
Samantaray S, Rout GR, Das P (1999) Studies on differential
tolerance of mungbean cultivars to metalliferous minewastes.
Agribiol Res 52:193201
Sing OV, Jain RK (2003) Phytoremediation of toxic aromatic
pollutants from soil. Appl Microbiol Biotechnol 63:128135
Singh N, Ma L Q (2003) Chinese brake fern, a potential
phytoremediator of arsenic contaminated soil and water.
International Society of Environmental Botanists, Vol 9,
No. 3
Shann JR (1995) The role of plants and plant/microbial systems
in the reduction of exposure. Environ Health Perspect 103
(suppl 5):0000
Suresh B, Ravishankar GA (2004) Phytoremediationa novel
and promising approach for environmental clean-up. Crit
Rev Biotechnol 24(23):97124
Suseela MR, Sinha S, Singh S, Saxena R (2002) Accumulation
of chromium and scanning electron microscope studies in
scirpus lacustris 1 treated with metal and tannery effluent.
Bull Environ Contam Toxicol 68:5408
United States Environmental Protection Agency (2001) A
citizens guide to phyoremediation. EPA 542-F-01-002.
Wenzel WW, Lombi E, Adriano DC (2004) Biogeochemical
processes in the rhizosphere: role in phytoremediation of
metal-polluted Soils. In: Prasad M, Hagemeyer J (eds)
Heavy metal stress in plants: from biomolecules to
ecosystems. Springer Verlag, Berlin Heidelberg New York,
pp 273303
Yi J-M, Chon H-T, Park M (2003) Migration and enrichment of
arsenic in the rock-soil-crop plant system in areas covered
with Black Shale, Korea. The Scientific World Journal
3:194205
Zayed A, Lytle CM, Qian J-H, Terry N (1998) Chromium
accumulation, translocation and chemical speciation in
vegetable crops. Planta 206:293299
Plant Soil (2010) 328:175189 189