Water Air Soil Pollut (2013) 224:1660

DOI 10.1007/s11270-013-1660-8

Plant Responses to Arsenic: the Role of Nitric Oxide

Fernanda S. Farnese & Juraci A. de Oliveira &
Grasielle S. Gusman & Gabriela A. Leo &
Cleberson Ribeiro & Luhan I. Siman & Jos Cambraia

Received: 25 February 2013 / Accepted: 16 July 2013 / Published online: 6 August 2013
# Springer Science+Business Media Dordrecht 2013

Abstract Arsenic (As) toxicity and the effects of nitric

oxide (NO), supplied as sodium nitroprusside (SNP),
were analyzed in Pistia stratiotes. The plants, which
were grown in nutrient solution at pH 6.5, were exposed
to four treatments for 24 h: control; SNP (0.1 mg L1);
As (1.5 mg L1); and As + SNP (1.5 and 0.1 mg L1). As
accumulated primarily in the roots, indicating the low
translocation factor of P. stratiotes. The As accumulation triggered a series of changes with increasing production of reactive oxygen intermediates and damage to
cell membranes. The application of SNP was able to
mitigate the harmful effects of As. This attenuation was
probably due to the action of the SNP as an antioxidant,
reducing the superoxide anion concentration, and as a
signaling agent. Acting as a signal transducer, SNP
increased the activity of enzymatic antioxidants (POX,
CAT, and APX) in the leaves and stimulated the entire
phytochelatins biosynthetic pathway in the roots (increased sulfate uptake and synthesis of amino acids,
F. S. Farnese : G. A. Leo
Department of Plant Biology, Federal University of Viosa,
Viosa, Brazil
J. A. de Oliveira (*) : C. Ribeiro : L. I. Siman : J. Cambraia
Department of General Biology,
Federal University of Viosa,
Viosa, Minas Gerais 36570-000, Brazil
e-mail: jalves@ufv.br
G. S. Gusman
College of Pharmacy, Federal University of Minas Gerais,
Belo Horizonte, Minas Gerais 31270-901, Brazil

non-proteinthiols, and phytochelatins). The As also

stimulated the phytochelatins biosynthesis, but this effect was limited, probably because plants exposed only
to pollutant showed small increments in the sulfate
uptake. Thus, NO also may be involved in gene regulation of sulfate carriers.
Keywords Antioxidants . Toxicity . Phytochelatins .
Cellular Signaling . Sulfate Uptake

1 Introduction
Arsenic (As) occurs naturally in the earths crust and
has been detected at toxic concentrations in water and
soil; primarily reflecting the anthropic actions. Due to
the high toxicity and increasing environmental concentrations of As, many research studies have been
conducted to control and/or remove this pollutant from
the environment. Among the techniques developed,
the use of accumulating plants has been highlighted
as a simple and viable solution (Yang et al. 2005).
Several plant species have effectively demonstrated
the phytoremediation of As-contaminated environments, and the aquatic macrophyte Pistia stratiotes L.
(Araceae) has been identified as a good candidate in
aquatic environments (Mufarrege et al. 2010).
The chemical form of As in surface water is arsenate
(AsV) (Mandal and Suzuki 2002), and because of its
chemical similarity to phosphate, plants easily absorb

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Water Air Soil Pollut (2013) 224:1660

AsV, causing direct and indirect cellular damage

(Gusman et al. 2013). The indirect toxic effects of this
absorption primarily result from the oxidative stress
caused by the increased production of reactive oxygen
intermediates (ROIs) (Zhang and Qui 2007).
The increased production of ROIs alters the normal
metabolism of plants and might cause cell death,
depending on the extent of ROI-induced damage
(Zhang and Qui 2007). However, plants have developed
mechanisms to mitigate these effects using enzymatic
antioxidants, such as superoxide dismutase (SOD), peroxidases (POX), catalases (CAT), ascorbate peroxidases
(APX), as well as non-enzymatic antioxidants, such as
glutathione and thiols (Singh et al. 2009). Another important strategy is the chelation of heavy metals. One of
the principal classes of heavy metal chelators known in
plants are phytochelatins (PCs), a family of Cys-rich
peptides and the most abundant class of non-protein
thiols (Zhang et al. 2012).
The activation of response mechanisms to heavy
metals involves a complex network of stimuli and
signaling molecules, like hormones and the nitric oxide
(NO) (Leitner et al. 2009). NO is a small molecule that
participates as a signal in several biochemical and
physiological processes in plants. Indeed, NO plays a
fundamental role in controlling physiological functions
during plant growth and development besides mediating plants responses to abiotic stresses, such as heavy
metal toxicity (Gonzlez et al. 2012). However, little
information is available concerning the role of NO in
the regulation of As-induced stress (Leterrier et al.
2012). Therefore, the aim of this study was to examine
the role of NO in the attenuation of As-induced oxidative stress in P. stratiotes.

irradiance (252 C; 230 mol m2 s1), under a photoperiod of 16 h, for an adaptation period of 3 days.
After the adaptation period, plants were transferred to
1.0-L polyethylene pots containing 0.5-L Clarks nutrient solution, pH 6.5, with 1/2 of the full ionic strength
and exposed to four treatments: control (nutrient solution
only), sodium nitroprusside (SNP) (0.1 mg L1), As
(1.5 mg L1) and As + SNP (1.5 and 0.1 mg L1),
respectively. Sodium nitroprusside is a substance that is
commonly used in biochemical studies as an NO donor.
The concentration of As chosen was the maximum concentration at which the plant still showed positive growth
rate (in higher concentrations the growth rate was negative due the loss of root system) and the concentration of
SNP selected was one in which the index of tolerance to
arsenic was approximately 50 %. The plants were
maintained under these conditions for 24 h.

2 Material and Methods

2.2 Concentration of Reactive Oxygen Intermediates

Specimens of P. stratiotes collected in non-polluted

dams at the Federal University of Viosa, Viosa,
Minas Gerais State, Brazil, were used in all experiments. Plants of similar size (about 4.0 g fresh weight)
were surface sterilized with 1 % sodium hypochlorite
for 1 min and then extensively rinsed with running tap
water and demineralized water, and maintained in
demineralized water for 24 h. Then, they were transferred to polyethylene flasks with 10 L of Clarks
nutrient solution, pH 6.5 (Clark 1975), and maintained
in a growth room with controlled temperature and

To determine the concentration of the superoxide anion

(O2), 50 mg of leaf samples were incubated in an
extraction medium consisting of 100-M ethylenediaminetetraacetic acid (EDTA) disodium salt, 20-M
NADH, and 20-mM sodium phosphate buffer, pH 7.8
(Mohammadi and Karr 2001). The reaction was initiated by adding 100 L of 25.2-mM epinephrine in
0.1-N HCl. The samples were incubated at 28 C under
stirring for 5 min. The absorbance was read at 480 nm
for 5 min. Superoxide anion production was assessed
by determining the accumulated adenochrome, using

2.1 Determination of Arsenic

The leaves and roots of P. stratiotes were separated,
washed in deionized water, and placed in a conventional oven at 80 C until a constant dry weight was
obtained. The dry plant material was crushed and
digested in a mixture of nitric acid and perchloric acid
(Marin et al. 1993), and the concentrations of As was
determined through inductively coupled plasma emission spectroscopy (Optima 3300 DV, Perkin-Elmer,
Norwalk, CT). The accuracy of the method was verified by analysis of certified reference materials. The
reference material, Lemna minor (an aquatic plant)
(BCR-670), from the National Institute of Standards
and Technology (Gaithersburg, MD, USA), was used
for As determination.

Water Air Soil Pollut (2013) 224:1660

the molar absorption coefficient of 4.0103 M1

(Boveris et al. 2002).
The hydrogen peroxide (H2O2) concentration was
determined using 200 mg of leaf samples that were
homogenized in an extraction medium consisting of
50-mM potassium phosphate buffer, pH 6.5, containing 1-mM hydroxylamine and centrifuged at 10,000g
for 15 min at 4 C. Subsequently, 50-L aliquots of the
supernatant were added to a reaction medium containing 100-M FeNH4SO4, 25-mM sulfuric acid,
250-M xylenol orange, and 100-mM sorbitol (Gay
and Gebicki 2000). The samples were kept in the dark
for 30 min, and the absorbance was read at 560 nm.
The H2O2 concentrations were estimated based on a
calibration curve prepared with H2O2 standards.
2.3 Cellular Damage
The cellular damage was evaluated through an assessment of cell membrane integrity, quantifying electrolyte leakage according to Lima et al. (2002). Leaf discs
and root apices were obtained after each treatment,
rinsed thoroughly in distilled water, and maintained
in 10 mL of distilled water in sealed flasks for 6 h at
room temperature. The electrolyte leakage was estimated from the electrical conductivity in the solution
containing the root apices and leaf discs using an
electrical conductivity meter (DM31, Digimed, Santo
Amaro, Brazil). The conductivity was expressed as a
percentage of the total conductivity measured after
incubating the vials at 90 C for 2 h.
2.4 Assessment of Enzymes Activity
of the Antioxidant System
To assess the activity of the antioxidants and metabolism enzymes, 0.3 g of the roots and leaf fresh matter
were homogenized in extraction medium comprising
0.1-M potassium phosphate buffer, pH 6.8, 0.1-mM
EDTA, 1-mM phenylmethanesulfonyl fluoride, and
1 % polyvinylpyrrolidone (Peixoto et al. 1999). The
homogenate was centrifuged at 12,000g for 15 min at
4 C. The resulting supernatant was used as a crude
extract for the assessment of SOD, POX, APX, and
CAT activities.
The SOD activity (SOD, EC 1.15.1.1) was measured as the inhibition of p-nitro tetrazolium photoreduction according to the method of Giannopolitis and
Ries (1977). The enzymatic activity was expressed in

Page 3 of 11, 1660

SOD units corresponding to the amount of enzyme

required to inhibit 50 % of the p-nitro blue tetrazolium
photoreduction (Beauchamp and Fridovich 1971).
The POX activity (POX, EC 1.11.1.7) was assessed
through the production rate of purpurogallin at 420 nm
according to the proposed method of Nakano and
Asada (1981) with a molar extinction coefficient of
2.47 mmol1 L cm1. The enzymatic activity was
expressed in micromoles purpurogallin min1 g1 fresh
weight (FW).
The APX activity (APX, EC 1.11.1.11) was assessed
as the rate of ascorbate oxidation at 290 nm (Nakano
and Asada 1981) using a molar extinction coefficient of
2.8 mmol1 L cm1. The enzymatic activity was
expressed in micromoles ascorbate min1 g1 FW.
The CAT activity (CAT, EC 1.11.1.6) was estimated
through the decomposition of H2O2 during the first minute of the reaction at 240 nm (Havir and McHale 1987)
using a molar extinction coefficient of 36 mol1 L cm1.
The enzymatic activity was expressed in micromoles
H2O2 min1 g1 FW.
2.5 Effect of Arsenic in Sulfate Uptake
Plants were transferred to 0.5-L polyethylene pots containing Clarks nutrient solution, pH 6.5. Uptake of
sulfate by the plants was assessed by Claassen and
Barbers depletion method (Claassen and Barber
1974). Twenty-four hours before evaluation of the
kinetic constants, the nutrient solution was renewed
at every 6 h to obtain the steady state uptake. Finally,
the nutrient solution was renewed again, and half of the
pots received 1.5 mg L1of As, adding then 1.0 mL of
carrier freeNa235SO4 (0.042 MBq mL1). After the
introduction of the plants, the absorption solution was
sampled periodically during a period of 12 h. The
amount of radioactivity in each aliquot, after addition
of 10 mL of a liquid scintillation cocktail was determined in a Beckman LS 6500 Scintillation System.
The kinetic parameters Km and Vmax and the ratio
Vmax/Km were estimated by graphic-mathematical approach suggested by Ruiz (1985).
2.6 Amino Acids Content
Amino acid content was determined by the method of
Moore and Stein (1948). 0.5 g of plant sample was
homogenized in 10 mL of 80 % ethanol. The homogenate was centrifuged 800 rpm for 10 min. One

1660, Page 4 of 11

milliliter of the extract was taken in the test tube and

1 mL of 0.1 N HCl was added to neutralize the sample.
To this, 1 mL of ninhydrin reagent was added and
heated for 20 min in a boiling water bath. Later, 5 mL
of the diluent solution was added and heated again in
water bath for 10 min. The test tubes were cooled and
read the absorbance at 570 nm in a spectrophotometer.
2.7 Glutathione, Thiols, and Phytochelatins Content
Extraction and estimation of total glutathione (GSH) was
performed according to Griffiths method (Griffith 1980).
The leaf and root tissue was homogenized in 5 % (m/v)
sulfosalicylic acid and the homogenate was centrifuged at
10,000g for 10 min. A volume of 1 ml of supernatant
was neutralized with 0.5 ml of potassium phosphate
buffer (pH 7.5). Total GSH content was measured by
adding 1 ml neutralized supernatant to a standard solution
mixture consisting of 0.5 ml of 0.1 M sodium phosphate
buffer (pH 7.5) containing 1-ml EDTA, 0.2 ml of 6-mM
5,5-dithiobis-2-nitrobenzoic acid, 0.1 ml of 2-mM
NADPH, and 0.1 ml of 1-U yeast GR Type III. Change
in absorbance was measured at 412 nm and followed at
252 C until the absorbance reached 0.5 U.
To assess the thiols content, 0.3 g of the roots and leaf
fresh matter were homogenized in extraction medium
comprising 0.1-M TrisHCl buffer, pH 8.0, 1-mM
EDTA, and 1 % ascorbic acid. The homogenate was
centrifuged at 10,000g for 10 min at 4 C. The
resulting supernatant was used for determination of total
Fig. 1 Concentration of arsenic in the leaves (black)
and roots (gray) of Pistia
stratiotes. Means followed
by the same uppercase letter,
between treatments for the
same organ, and by the same
lowercase letter, between
organs for the same treatment, were not significantly
different according to
Tukeys test at 5 %
probability

Water Air Soil Pollut (2013) 224:1660

thiols, protein thiols, and non-protein thiols (Sedlak and

Lindsay 1968). The estimated concentration of total PCs
was calculated as PCs = non-protein thiols total GSH
(Hartley-Whitaker et al. 2001).
2.8 Statistical Analyses
The experiment was conducted as a completely randomized design with five replicates. The data were
analyzed using ANOVA, and the means were calculated using Tukeys test at 5 % probability. The statistical
analyses were conducted with the statistical program
SAS (Cary, NC, United States).

3 Results
3.1 Plant Arsenic Concentration and Translocation
Factor
The plants exposed to As accumulated large amounts
of the pollutant, and no changes were observed after
the addition of SNP (Fig. 1). Arsenic accumulation
occurred mainly in the roots with little translocation
to the leaves (Fig. 1).
3.2 Concentration of Reactive Oxygen Intermediates
In contrast to the values observed in the control, As
exposure increased the O2 concentration in the leaves

Water Air Soil Pollut (2013) 224:1660

Page 5 of 11, 1660

Fig. 2 Effect of arsenic, alone or in combination with SNP, on

the concentration of superoxide anion (a) and hydrogen peroxide
(b) in the leaves (black) and roots (gray) of Pistia stratiotes. Means
followed by the same uppercase letter, between treatments for the

same organ, and by the same lowercase letter, between organs for
the same treatment, were not significantly different according to
Tukeys test at 5 % probability

and roots of P. stratiotes, and SNP treatment had no

effect on the concentrations of this anion in the leaves
or roots (Fig. 2a). The application of SNP in combination with As, reduced the anion O2 concentration
approximately 31.8 and 10.7 % in the leaves and roots,
respectively, compared with plants exposed to As only.
The H2O2 concentration was also increased in the
leaves and roots of plants after exposure to As, and this
increase was higher in the roots (Fig. 2b). The SNP
treatment alone had no effect on the H2O2 concentration.
However, the application of this exogenous source of NO

(SNP) to As-treated plants induced a 28.5 and 21.7 %

reduction of the H2O2 concentration in the leaves and
roots, respectively.

Fig. 3 Electrolyte leakage

in the leaves (black) and
roots (gray) of Pistia
stratiotes. Means followed
by the same uppercase letter,
between treatments for the
same organ, and by the same
lowercase letter, between
organs for the same treatment, were not significantly
different according to
Tukeys test at 5 %
probability

3.3 Electrolyte Leakage

Plant treatments with As exhibited increased electrical
conductivity, which is indicative of increased electrolyte
leakage (Fig. 3). There was no difference between the
damage generated in the roots and leaves, despite the
elevated As concentration in the roots. Treatment with

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Water Air Soil Pollut (2013) 224:1660

Fig. 4 Activities of SOD (a), POX (b), APX (c), and CAT (d) in
the leaves (black) and roots (gray) of Pistia stratiotes. Means
followed by the same uppercase letter, between treatments for

the same organ, and by the same lowercase letter, between

organs for the same treatment, were not significantly different
according to Tukeys test at 5 % probability

SNP alone had no effect on electrolyte leakage; however,

when added to plants in combination with As, this treatment attenuated the As damage in the leaves and roots.

activity than that observed in plants exposed only to As

(Fig. 4bd). The increment in the activity of these
enzymes was much more intense in leaves. In fact,
the greatest increase in enzymatic activity was observed

3.4 Antioxidant Enzyme Activity

The activities of SOD, CAT, and POX significantly
increased in the leaves of As-treated plants (Fig. 4).
However, only increased SOD activity was observed in
the roots of plants exposed to this pollutant (Fig. 4a).
The application of SNP alone had no effect on enzyme
activity compared with the control. However, the Astreated plants exposed to SNP maintained SOD activity
similar to the control. In contrast, the activities of POX,
APX, and CAT were increased after the combined
treatment with As and SNP, showing higher enzymatic

Table 1 Estimated values of Vmax (in micromoles per hour per

grain fresh weight) and Km (in mole per liter) for absorption of
sulfate in Pistia stratiotes
Treatments

Vmax (mol h1 g1 FW)

Km (mol L1)

Control

0.13 c

9.78 a

SNP

0.11 c

11.3 a

As

0.19 b

8.61 a

As + SNP

0.27 a

8.93 a

Means followed by the same letter were not significantly different according to Tukeys test at 5 % probability

Water Air Soil Pollut (2013) 224:1660

Page 7 of 11, 1660

Fig. 5 Effect of arsenic,

alone or in combination
with SNP, on the concentration of total glutathione in
the leaves (black) and roots
(gray) of Pistia stratiotes.
Means followed by the same
uppercase letter, between
treatments for the same organ, and by the same lowercase letter, between organs
for the same treatment, were
not significantly different
according to Tukeys test at
5 % probability

with CAT in the leaves of plants exposed to As alone or

in combination with SNP. However, the CAT activity
remained unaltered in the roots of these plants.

3.5 Effect of Arsenic in Sulfate Uptake

The kinetic constants of sulfate uptake were changed after
As treatment. The presence of the metalloid in combination with SNP, however, intensified the increase of Vmax
for sulfate, but had no significant effect on Km (Table 1).
Fig. 6 Effect of arsenic,
alone or in combination with
SNP, on the concentration of
amino acid in the leaves
(black) and roots (gray) of
Pistia stratiotes. Means
followed by the same uppercase letter, between
treatments for the same organ, and by the same lowercase letter, between organs
for the same treatment, were
not significantly different
according to Tukeys test at
5 % probability

3.6 Effect of Arsenic in Glutathione, Amino Acids,

Thiols, and Phytochelatins
The concentrations of GSH were decreased by As
(Fig. 5), while the levels of amino acids (Fig. 6), total
thiols, non-protein thiols (Fig. 7a and b), and PCs (Fig. 8)
increased. The application of SNP intensified all the
responses in roots, inducing changes even more significantly. In the leaves, the concentration of GSH and PCs
were decreased while the concentration of protein thiols
increased after the application of SNP (Fig. 7c).

1660, Page 8 of 11

Fig. 7 Concentration of total thiols (a), non-protein thiols (b), and

protein thiols (c) in the leaves (black) and roots (gray) of Pistia
stratiotes. Means followed by the same uppercase letter, between

Fig. 8 Effect of arsenic,

alone or in combination with
SNP, on the concentration of
phytochelatins in the leaves
(black) and roots (gray) of
Pistia stratiotes. Means
followed by the same uppercase letter, between
treatments for the same organ, and by the same lowercase letter, between organs
for the same treatment, were
not significantly different
according to Tukeys test at
5 % probability

Water Air Soil Pollut (2013) 224:1660

treatments for the same organ, and by the same lowercase letter,
between organs for the same treatment, were not significantly
different according to Tukeys test at 5 % probability

Water Air Soil Pollut (2013) 224:1660

4 Discussion
The results of the present study suggest that the macrophyte P. stratiotes L. (Araceae), compared with other
aquatic species, has great potential for the phytoremediation of aquatic environments polluted with As.
Besides the large accumulation of As, it was observed
that P. stratiotes has high tolerance to this pollutant.
Most of the absorbed As was retained in the roots of
P. stratiotes, as demonstrated by a translocation factor
value lower than 1, indicating great retention in the
roots and low metalloid translocation to the shoots. The
translocation factor depends not only on the plant
species but also on the absorbed toxic element. P.
stratiotes plants exposed to Hg, for example, showed
a translocation factor of four times greater than that
observed with As (Mishra et al. 2009). The ability to
maintain the toxic element in the roots is considered as
a key mechanism of adaptation to contaminated areas
(Mishra et al. 2009) and typically reflects the presence
of molecules rich in SH groups, which chelate As and
prevent the translocation of this pollutant to the leaves
(Singh and Agrawal 2007). In fact, high concentrations
of thiols were observed in roots.
Given the difficulty of removing the roots from the
soil, pollutant retention in the roots might be problematic
for the phytoremediation of land environments. However, in aquatic environments, this problem is minimized
through the removal of the entire plant from the environment. Furthermore, the roots are more tolerant to As than
the leaves, despite showing higher pollutant concentrations, suggesting that changes in lipid peroxidation in the
roots are lower than those observed in the shoots.
The P. stratiotes plants exposed to As experienced
oxidative stress, as demonstrated by the increase in ROIs
concentration and membrane damage, causing electrolyte leakage. The increase in ROIs production might
reflect the conversion of arsenate into arsenite, which is
part of the mechanism of pollutant tolerance (Meharg
and Hartley-Whitaker 2002) through the direct reduction
of molecular oxygen or the alteration of proteins in the
mitochondria, chloroplasts, and peroxisomes (Zhang and
Qui 2007). The toxic effect of As, however, was attenuated after the addition of NO (supplied as SNP), which
acted as both a direct antioxidant, eliminating O2, and as
a signal transducer, enhancing the response of enzymatic
antioxidants and the sulfate uptake.
With certain types of abiotic stress, SNP acts as a
cellular signal transducer to increase SOD activity and

Page 9 of 11, 1660

thereby reduces the concentration of O2 (Arasimowicz

and Floryszak-Wieczorek 2007). In P. stratiotes, however, no significant changes in SOD activity were observed in the presence of As and SNP, suggesting that
SNP acted directly as an antioxidant (Singh et al. 2009;
Xiong et al. 2010). For CAT, POX, and APX, the NO
released from SNP acted as a signal transducer (Singh
et al. 2009), inducing an increase in the enzyme activity
and a reduction in the H2O2 concentration. These increases were much more significant in the leaves than
that in the roots.
The activity of antioxidant enzymes is essential to
eliminate the ROIs, which are an indirect and harmful
effect of absorption of As. However, in addition to
repairing the damage caused by As, plants resistant to
pollutants also must provide mechanisms to inactivate
the toxic compound, thus preventing further damage
(Zhang et al. 2012). The main strategy for As detoxification in plant cells is based on chelation by PCs and
subsequent compartmentalization of the AsPCs complex (Zhang et al. 2012).
PCs are the most abundant class of non-protein thiols
and their synthesis involves the participation of amino
acids and glutathione (Zhang et al. 2012). In the present
investigation, the increases in the levels of amino acids,
non-protein thiols and PCs, as well as depletion of GSH
pools, suggests a stimulation of the entire PC biosynthetic pathway by the NO in the roots. These results
were not observed in the leaves, where the major mode
of action of SNP was the increment in enzyme activity.
Although the As also has been able to stimulate the PCs
biosynthetic pathway in the roots and in the leaves, the
observed increase was limited and was not able to
prevent the damage triggered by the pollutant. The
defense mechanism involving PCs results in an increasing demand for sulfur, making necessary increases in
sulfate uptake (Nocito et al. 2006). The plants exposed
only to As showed small increments in sulfate uptake,
which was probably the main factor that limited the
synthesis of PCs. Transcriptional regulation of the genes
encoding high-affinity sulfate transporters is linked to
the availability of sulfate, the demand for reduced sulfur
compounds, and the supply of a C/N skeleton precursor
needed in the assimilatory pathway (Nocito et al. 2006).
Our results suggest that NO may be involved in gene
regulation of carriers sulfate, since the NO was able to
increase the uptake of this ion.
Taken together, this data suggests that exogenous NO
mitigates As-induced damage in P. stratiotes plants. The

1660, Page 10 of 11

beneficial effects of NO most likely reflect the direct

elimination of ROIs, the increased activity of enzymes
in the antioxidant system, and the stimulation of the PC
biosynthetic pathway. In the latter case, NO is probably
involved in the up-regulation of genes involved in the
absorption of sulfate.

Acknowledgments The authors are grateful to the Universidade

Federal de Viosa, CNPq, and FAPEMIG.

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