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Rice Science, 2017, 24(3): 155í162

Antioxidant Defense Mechanisms of Salinity Tolerance in Rice

Genotypes

Mohammad Golam KIBRIA1, Mahmud HOSSAIN1, Yoshiyuki MURATA2, Md. Anamul HOQUE1
(1Department of Soil Science, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh; 2Graduate School of Natural
Science and Technology, Okayama University, Okayama 700-8530, Japan)

Abstract: In order to elucidate the role of antioxidant responses in salinity tolerance in rice genotypes
under salt stress, experiments were conducted using four rice varieties, including salt-sensitive BRRI
dhan 28 and three salt-tolerant varieties BRRI dhan 47, BINA dhan 8 and BINA dhan 10. Thirty-day-old
rice seedlings were transplanted into pots. At the active tillering stage (35 d after transplanting), plants
were exposed to different salinity levels (0, 20, 40 and 60 mmol/L NaCl). Salt stress caused a significant
reduction in growth for all the rice genotypes. Growth reduction was higher in the salt-sensitive genotype
than in the salt-tolerant ones, and BINA dhan 10 showed higher salt tolerance in all measured
physiological parameters. The reduction in shoot and root biomass was found to be minimal in BINA dhan
10. Chlorophyll content significantly decreased under salt stress except for BINA dhan 10. Proline content
significantly increased in salt-tolerant rice genotypes with increased salt concentration, and the highest
proline content was obtained from BINA dhan 10 under salt stress. Catalase and ascorbate peroxidase
activities significantly decreased in salt-sensitive genotype whereas significantly increased in salt-tolerant
ones with increasing salt concentration. However, salt stress significantly decreased guaiacol peroxidase
activity in all the rice genotypes irrespective of salt tolerance. K+/Na+ ratio also significantly decreased in
shoots and roots of all the rice genotypes. The salt-tolerant genotype BINA dhan 10 maintained higher
levels of chlorophyll and proline contents as well as catalase and ascorbate peroxidase activities under
salt stress, thus, this might be the underlying mechanism for salt tolerance.
Key words: antioxidant enzyme; salinity; chlorophyll content; proline content; K+/Na+ ratio; rice

Climate change is a serious environmental threat that
causes sea level rise and thereby affects coastal areas
of Bangladesh. Salinity is one of the major abiotic
factors, limiting crop production worldwide.
Approximately 7% of the world’s total land area, 20%
of the world’s cultivated land area and nearly 50% of
the world’s irrigated land area are affected by salinity
(Zhu, 2001). In Bangladesh, about 1.06 million
hectares of arable lands are affected by soil salinity
(SRDI, 2010). Salinity is a serious threat to rice
production in the southern part of Bangladesh.
Salinity imposes both ionic toxicity and osmotic stress
to plants (Hasegawa et al, 2000; Zhu, 2003). Salt
stress disturbs cytoplasmic K+/Na+ homeostasis,
causing a decrease in K+/Na+ ratio in the cytosol (Zhu,
2003). Accumulation of excess Na+ and Cl– causes
ionic imbalance that may impair the selectivity of root
membranes and induce K+ deficiency in plants.
Rice is the most important cereal crop in the world,
yielding one-third of the total carbohydrate sources.
The total rice growing area in Bangladesh is about
10.83 million hectares, leading to the production of
33.54 million metric tons of rice. On the contrary, rice
yield is very low in the salt soil. To increase rice yield
per acre, it is imperative to know about physiological
and biochemical changes in plants under salt stress.

Received: 28 September 2016; Accepted: 29 December 2016

Corresponding author: Md. Anamul HOQUE (anamul71@yahoo.com)
Copyright © 2017, China National Rice Research Institute. Hosting by Elsevier B V This is an open access article under the CC BY-NC-ND license
B.V.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Peer review under responsibility of China National Rice Research Institute
http://dx.doi.org/
http://dx.doi.org/10.1016/j.rsci.2017.05.001
156
Plants have evolved a variety of adaptive mechanisms
to respond to salt stress. One of the main adaptive
mechanisms is the accumulation of compatible solutes
(Sharma and Dietz, 2006; Ashraf and Foolad, 2007),
and proline is the most common one. Increased levels
of endogenous proline accumulation in plants is
correlated with enhanced salt tolerance (Hasegawa et al,
2000; Ashraf and Foolad, 2007; Boscaiu et al, 2012;
Sripinyowanich et al, 2013), and induce oxidative
stress tolerance by modulating the activities of
antioxidant enzymes (Saeedipour, 2013). Proline
accumulation also plays a regulatory role during plant
growth under salt stress (Mattioli et al, 2008).
Salinity induces the production of reactive oxygen
species (ROS) in plant cells (Hasegawa et al, 2000;
Banu et al, 2009, 2010). ROS can act as signaling
molecules, mediating many key physiological processes.
Excess production of ROS is toxic to plants and causes
oxidative damage to cellular constituents, leading to
cell death (Noctor and Foyer, 1998; Banu et al, 2009,
2010). Plants possess enzymatic and non-enzymatic
antioxidant defense systems to protect cells against the
damaging effects of ROS. The major ROS-scavenging
antioxidant enzymes are catalase (CAT), guaiacol
peroxidase (POX) and ascorbate peroxidase (APX).
Salt stress shows different effects on the components
of antioxidant defense systems in plants (Hasegawa et al,
2000; Mittova et al, 2003; Demiral and Turkan, 2005;
El-Shabrawi et al, 2010; Hasanuzzaman et al, 2014).
There is increasing evidence that salt stress has a
significant effect on physiological and biochemical
attributes of plants. Better understanding of physiological
and biochemical characteristics of plants is vital for
improving salt tolerance under salinity. To clarify the
physiological and biochemical responses of salt-sensitive
and salt-tolerant rice genotypes, we investigated the
plant growth, chlorophyll content, proline content and
activity of antioxidant enzymes in four contrasting
rice genotypes exposed to salt stress, which can be
used as index for in vitro salt tolerance in rice.
MATERIALS AND METHODS
Rice materials and experimental design
Four rice genotypes, including one salt-sensitive
variety (BRRI dhan 28) and three salt-tolerant
varieties (BRRI dhan 47, BINA dhan 8 and BINA
dhan 10), were exposed to different NaCl levels (0, 20,
40 and 60 mmol/L). The experiment was laid out in a
Rice Science, Vol. 24, No. 3, 2017
randomized complete block design with four
replications. Considerable spacing was maintained
among the pots for the ease of management practices.
Pot preparation and management practices
Pot experiments were carried out at a net-house
(average temperature 24 ºC and relative humidity 60%)
of the Department of Soil Science, Bangladesh
Agricultural University, Bangladesh to investigate the
effects of salt stress on the growth as well as the
physiological and biochemical characteristics of
salt-sensitive and salt-tolerant rice.
Soils were collected from the Soil Science Field
Laboratory, Bangladesh Agricultural University. The
initial soil sample was silt loam having exchangeable
Na 0.383 meq/100 g and exchangeable K 0.082
meq/100 g. Each 15 L plastic pot was filled with 8 kg
soil and 5 L water, ensuring enough space to maintain
flooded conditions. Thirty-day-old seedlings were
transplanted into pots. No NaCl was added to soils for
control treatment. For 20, 40 and 60 mmol/L NaCl
treatments, 15.24, 30.47 and 45.71 g pure NaCl were
dissolved in 1 000 mL water and then added to the
pots, respectively, at the active tillering stage (five
weeks after transplanting). The soil in the pot was
flooded with salt solution and the EC values of soil for
control, 20, 40 and 60 mmol/L NaCl treatments were
1.00, 2.25, 5.60 and 6.58 dS/m, respectively. The
crops were harvested at two weeks after salt treatment.
Whole plants (with root attached) were carefully
uprooted from the soil so that the root system of the
plants remained unaffected.
Plant height, root length, shoot fresh weight, shoot
dry weight, root fresh weight, root dry weight and
number of tillers per hill were recorded.
Biochemical analysis
Assay of chlorophyll content
Chlorophyll content was measured according to Porra
et al (1989). An aliquot of fresh leaf (0.5 g) was
suspended in 10 mL of 80% acetone, mixed well and
kept at room temperature in the dark for 7 d. The
supernatant was collected after centrifugation at 5 000
r/min for 15 min. The sample absorbance was recorded
at 645 and 663 nm using a spectrophotometer (Model
336001, Spectronic Instruments, USA).
Assay of proline content
Proline content was measured according to Bates et al
(1973). An aliquot of fresh leaf (0.5 g) was
Mohammad Golam KIBRIA, et al. Antioxidant Defense Mechanisms of Salinity Tolerance in Rice 157

homogenized in 10 mL of 3% sulfosalicylic acid, and Statistical analysis

the homogenate was centrifuged at 5 000 r/min for 15
min. A total of 2 mL supernatant was incubated with 2
mL acid ninhydrin (1.25 g ninhydrin dissolved in 30
mL glacial acetic acid and 20 mL of 6 mol/L
phosphoric acid) and 2 mL glacial acetic acid for 1 h
at 100 ºC, and then quickly cooled in an ice bath. The
colored reaction mixture was extracted with 4 mL
toluene, and the absorbance was recorded at 520 nm.
Preparation of enzyme extract and assay of
antioxidant enzymes
An aliquot fresh leaf (0.5 g) was homogenized with 5
mL of 50 mmol/L Tris-HCl buffer (pH 8.0) for CAT
and 50 mmol/L KH2PO4 buffer (pH 7.0) for POX and
APX. The homogenate was centrifuged at 5 000 r/min
for 20 min, and the supernatant was then used for
enzyme activity assays. CAT (EC: 1.11.1.6) activity
was assayed as described by Islam et al (2009). POX
(EC: 1.11.1.7) and APX (EC: 1.11.1.11) activities
were assayed as described by Hoque et al (2007a, b).
Determination of K+ and Na+
Grain and straw samples were dried in an oven at
about 65 ºC for 48 h, and then samples were ground in
a grinding machine to pass through a 20-mesh sieve.
Ground sample (0.5 g) was added to 10 mL di-acid
mixture (HNO3:HClO4 = 2:1) in a 100 mL digestion
vessel. After a brief incubation at room temperature,
the vessels were heated slowly to 200 ºC. Heating was
stopped when a dense white fume of HClO4 was
observed. After cooling, the digested tissue was
transferred into a 50 mL volumetric flask and brought
to a final volume of 50 mL with distilled water. K+
and Na+ contents were determined using a flame
photometer (Jencon PFP 7, JENCONS-PLS, UK)
following to Brown and Lilleland (1946).
Data were analyzed using the statistical package
MStatC. The mean differences were adjudged by
Duncan’s multiple range test (Gomez and Gomez,
1984).
RESULTS
Root and shoot growth
Salt stress caused significant reductions in shoot and
root growth of all the rice genotypes (Table 1). Except
salt-sensitive genotype, all salt-tolerant rice genotypes
are survived at the highest NaCl level (60 mmol/L).
BINA dhan 10 showed 100% survival and growth at
60 mmol/L NaCl stress whereas the other tolerant
genotypes BRRI dhan 47 and BINA dhan 8 displayed
tolerance up to 40 mmol/L NaCl. The sensitive
genotype (BRRI dhan 28) survived only at low salinity
(20 mmol/L NaCl). At 60 mmol/L NaCl treatment, the
highest fresh and dry weights of root and shoot were
obtained in BINA dhan 10, followed by BRRI dhan 47
and BINA dhan 8. BINA dhan 10 showed reduction in
shoot fresh weight by 12%, 19%, 46% and shoot dry
weight by 2%, 26% and 48% at 20, 40 and 60 mmol/L
NaCl, respectively. In the salt-sensitive genotype
BRRI dhan 28, shoot fresh weight was reduced by
61% and shoot dry weight was reduced by 60% at 40
mmol/L NaCl. The reduction in root fresh weight was
the highest in BRRI dhan 28 (77%) and the lowest in
BINA dhan 10 (57%) at 60 mmol/L NaCl treatment.
Plant height, root length and number of tillers per
hill
Soil salinity also caused significant reductions in plant

Table 1. Effects of salinity on fresh weight and dry weight in salt-sensitive and salt-tolerant rice genotypes. g
Shoot fresh weight Shoot dry weight
Treatment
BRRI dhan 28 BRRI dhan 47 BINA dhan 8 BINA dhan 10 BRRI dhan 28 BRRI dhan 47 BINA dhan 8 BINA dhan 10
Control 43.12 a 32.56 a 30.29 a 28.78 a 10.71 a 7.57 a 7.43 a 5.67 a
20 mmol/L NaCl 29.84 b 26.57 b 23.78 b 25.42 b 7.07 b 6.73 b 4.83 b 5.53 a
40 mmol/L NaCl 16.72 c 18.20 c 16.76 c 23.36 b 4.32 c 4.53 c 3.79 c 4.20 b
60 mmol/L NaCl 6.32 d 14.22 d 12.43 d 15.45 c 1.43 d 2.86 d 2.51 d 2.96 c
SE 0.63 0.64 0.16 0.22 0.33 0.15 0.16 0.13
Root fresh weight Root dry weight
Treatment
BRRI dhan 28 BRRI dhan 47 BINA dhan 8 BINA dhan 10 BRRI dhan 28 BRRI dhan 47 BINA dhan 8 BINA dhan 10
Control 19.58 a 19.05 a 18.32 a 17.09 a 7.49 a 5.64 a 6.39 a 5.92 a
20 mmol/L NaCl 16.63 b 17.58 b 12.46 b 15.40 b 4.46 b 5.08 b 4.62 b 4.88 b
40 mmol/L NaCl 11.76 c 13.57 c 10.86 c 10.21 b 2.61 c 2.59 c 3.12 b 3.22 c
60 mmol/L NaCl 4.45 d 6.29 d 6.82 d 7.28 c 1.73 d 1.54 d 1.78 c 1.91 d
SE 0.36 0.29 0.41 0.29 0.11 0.11 0.11 0.14
The means with the same letter in a column show insignificant difference at the 0.05 level (n = 4).
158 Rice Science, Vol. 24, No. 3, 2017

Table 2. Effects of salinity on plant traits in different rice genotypes.

Plant height (cm) Root length (cm) Number of tillers per hill
Treatment BRRI BRRI BINA BINA BRRI BRRI BINA BINA BRRI BRRI BINA BINA
dhan 28 dhan 47 dhan 8 dhan 10 dhan 28 dhan 47 dhan 8 dhan 10 dhan 28 dhan 47 dhan 8 dhan 10
Control 62.5 a 66.5 a 65.5 a 64.5 a 27.5 a 25.5 a 26.5 a 27.0 a 13 a 13 a 15 a 16 a
20 mmol/L NaCl 60.0 b 63.0 b 61.0 b 63.5 a 17.5 b 21.0 b 18.5 b 22.5 b 12 b 12 b 14 a 15 b
40 mmol/L NaCl 51.0 c 60.0 c 55.5 c 54.0 b 14.5 c 15.5 c 16.5 c 18.0 c 8c 9c 10 c 12 c
60 mmol/L NaCl 45.0 d 58.5 c 52.0 d 49.0 c 12.5 d 14.0 c 13.0 d 16.0 c 6d 8d 8d 11 d
SE 0.61 0.45 0.50 0.36 0.45 0.50 0.54 0.62 0.34 0.27 0.34 0.26
The means with the same letter in a column show insignificant difference at the 0.05 level (n = 4).

height, root length and number of tillers per hill in all
the rice varieties. Plant height significantly decreased
in all the varieties with increasing salinity levels. At
the highest salt stress (60 mmol/L NaCl), the tallest
plants (58.5 cm) were obtained in BRRI dhan 47 and
the shortest plants (45.0 cm) in BRRI dhan 28 (Table
2). At 20 mmol/L NaCl, root length was significantly
decreased in all the rice genotypes compared to the
control, but no significant differences were observed
between 40 and 60 mmol/L NaCl treatments in BINA
dhan 10 and BRRI dhan 47. The salt-sensitive
genotype showed a significant decrease in number of
tillers per hill at 20 mmol/L NaCl treatment. The
highest number of tillers per hill was obtained in
BINA dhan 10 among the four rice genotypes
irrespective of salinity level.
Chlorophyll content
Chlorophyll a content in rice showed different responses
to salinity stress. The salt-sensitive rice genotype
BRRI dhan 28 showed a decrease in chlorophyll a
content at 40 and 60 mmol/L NaCl. In salt-tolerant
rice genotypes, chlorophyll a content increased with
salinity level up to 40 mmol/L NaCl, but decreased at
60 mmol/L NaCl (Table 3). Chlorophyll b content
decreased in all the rice genotypes with increasing salt
concentrations. A reduction in total chlorophyll
content was observed with increasing salt concentration
in all the genotypes but this reduction was more
pronounced in the salt-sensitive genotype than the
tolerant ones. Compared to the control, BRRI dhan 28,
BRRI dhan 47 and BINA dhan 8 showed reduction of
total chlorophyll content by 38%, 32% and 42% at 60
mmol/L NaCl, respectively. It is important to note that
only 2% reduction in total chlorophyll content was
recorded in BINA dhan 10 at 60 mmol/L NaCl
treatment.
Proline content
The proline content in all the salt-tolerant genotypes
increased with increasing salinity levels (Fig. 1).
However, proline content in the salt-sensitive genotype
decreased significantly at 60 mmol/L NaCl. At 60
mmol/L NaCl, the highest proline content was obtained
in BINA dhan 10 whereas the lowest content was
found in salt-sensitive rice (BRRI dhan 28). BINA
dhan 10 accumulated approximately 2.2-fold higher
proline while BRRI dhan 47 and BINA dhan 8
accumulated 1.7- and 1.4-fold higher proline as
compared to the control, respectively.
Activities of antioxidant enzymes
To investigate whether soil salinity influenced the
antioxidant defense system in rice, the activities of
major ROS-scavenging antioxidant enzymes, including
CAT, POX and APX, were measured. Activities of
antioxidant enzymes were significantly affected in
response to salinity stress. CAT and APX activities in
salt-tolerant genotypes increased with increasing
salinity levels (Fig. 1). POX activities in all the rice
genotypes showed a decreasing trend with increasing
salt concentrations irrespective of salt tolerance levels

Table 3. Effects of salinity on chlorophyll content in different rice genotypes. mg/g

Chlorophyll a Chlorophyll b Total chlorophyll
Treatment BRRI BRRI BINA BINA BRRI BRRI BINA BINA BRRI BRRI BINA BINA
dhan 28 dhan 47 dhan 8 dhan 10 dhan 28 dhan 47 dhan 8 dhan 10 dhan 28 dhan 47 dhan 8 dhan 10
Control 4.53 a 3.88 c 4.18 b 4.08 b 5.19 a 5.48 a 5.16 a 4.98 a 9.72 a 9.36 a 9.37 a 9.06 a
20 mmol/L NaCl 4.56 a 4.01 b 4.21 b 4.10 b 5.02 a 4.97 b 4.95 a 4.90 b 9.61 a 8.98 b 9.16 a 9.00 a
40 mmol/L NaCl 4.11 b 4.42 a 4.34 a 4.21 a 4.22 b 3.62 c 4.26 b 4.69 c 8.33 b 8.04 c 8.60 b 8.90 a
60 mmol/L NaCl 3.60 c 3.67 d 3.44 c 4.19 a 2.38 c 2.69 d 1.96 c 4.68 c 5.98 c 6.36 d 5.40 c 8.87 a
SE 0.19 0.17 0.11 0.03 0.22 0.33 0.27 0.07 0.40 0.30 0.34 0.21
The means with the same letter in a column show insignificant difference at the 0.05 level (n = 4).
Mohammad Golam KIBRIA, et al. Antioxidant Defense Mechanisms of Salinity Tolerance in Rice 159

CAT activity [mmol/(min·g)]

POX activity [ȝmol/(min·g)]

APX activity [ȝmol/(min·g)]

Fig. 1. Effects of salinity on proline content, enzyme activities and K+/Na+ ratio in different rice genotypes (Mean ± SD, n = 4).
CAT, Catalase; POX, Guaiacol peroxidase; APX, Ascorbate peroxidase.
Means with different letters are significantly different at the 0.05 level.

(Fig. 1). CAT and APX activities in salt-sensitive rice approximately one-third (shoot) and half (root) of that
decreased with increased salinity. At any given in control plants in the salt-sensitive genotype.
concentration of NaCl, the decrease in enzyme
activities was more pronounced in the salt-sensitive DISCUSSION
rice genotype than the salt-tolerant ones.
K+/Na+ ratio in rice Effects of salinity on rice growth

K+/Na+ ratio in the shoots and roots of all the rice
genotypes significantly decreased due to salt stress
(Fig. 1). BINA dhan 10 showed a higher K+/Na+ ratio
compared to the other salt-tolerant genotypes. In
plants treated with 60 mmol/L NaCl, K+/Na+ ratio was
Reduced growth is a common phenomenon when
plants are grown in saline conditions. Inhibition of
growth due to salt stress has been observed even in
tolerant plant species (Mittler et al, 2001). Rice is
considered as a salt-sensitive crop (Shereen et al, 2005;
160
Darwish et al, 2009). In this study, significantly higher
growth was observed in salt-tolerant genotype BINA
dhan 10. Inhibitions of shoot and root growth are the
most important agricultural indices of salt tolerance
(Tuna et al, 2008). The present study demonstrated
that shoot and root growth significantly decreased due
to increased level of salinity (Table 1). Shannon and
Grieve (1998) also reported decreased shoot and root
biomass of rice under increased salt stress. The results
from this study also indicated significant declines in
plant height, root length and number of tillers per hill
as concentration of NaCl increased (Table 2). Similar
results in rice as well as in other crops have been
reported (Dadkhah et al, 2001; Momayezi et al, 2010).
Effects of salinity on chlorophyll content
Chlorophyll is one of the most important plant
pigments, supporting photosynthetic ability.
Chlorophyll content can vary due to salinity level,
eventually affecting plant growth and development. A
reduction in plant chlorophyll content was observed
with increasing salt concentration in all the genotypes,
however, the reduction was more pronounced in the
salt-sensitive genotype than in the salt-tolerant ones
(Table 3). The concentration of the pigment fractions
(chlorophyll a and chlorophyll b) in the leaves of all
the rice varieties clearly demonstrated that the
biosynthesis of photosynthetic pigments was affected
by NaCl stress (Table 3). These results are in
agreement with those of Islam et al (2009) and Abeer
et al (2013) in rice. The decrease in chlorophyll
content is an indicative response across different
plants subjected to salinity stress (Roy and Basu,
2008). Parida and Das (2005) suggested that the
decrease in chlorophyll content in response to salt
stress is a widespread phenomenon. Chen and Yu
(2007) also observed a significant decrease in
chlorophyll content at high NaCl level.
Effects of salinity on proline content
Increased accumulation of proline in plants is
correlated with improved salinity tolerance (Ashraf
and Foolad, 2007; Hasanuzzaman et al, 2014). Proline
content significantly increased in all the genotypes
with increasing salt concentration (Fig. 1). Among the
salt-tolerant rice genotypes, BINA dhan 10
accumulated 2.2-fold higher proline while the other
genotypes BINA dhan 8 and BRRI dhan 47
accumulated 1.5- and 1.7-fold higher proline at 60
mmol/L NaCl compared to the control, respectively.
Rice Science, Vol. 24, No. 3, 2017
The proline accumulation was also higher at 20 and 40
mmol/L NaCl in BRRI dhan 28, however, the
accumulation decreased at a higher level of salinity
(60 mmol/L NaCl) (Fig. 1). The decrease in proline
accumulation in the salt-sensitive rice genotype was
observed probably due to low synthesis of proline or
higher degradation of proline under high salinity stress.
In many studies, a positive correlation between the
accumulation of proline and stress tolerance in plants
has been found (Lutts et al, 1996; Kumar et al, 2003).
Higher proline content in BINA dhan 10 might be one
reason for the observed higher salt tolerance when
compared to the other genotypes. Demiral and Turkan
(2005) also reported that enhanced salt tolerance of
rice is correlated with increased capacity of
antioxidant system. Proline has been suggested to
function as an antioxidant in protecting cells against
various abiotic stress, since proline scavenges free
radicals and suppresses ROS accumulation.
Effects of salinity on antioxidant enzyme activities
Increased plant antioxidant response has been shown
to be positively associated with decreased oxidative
damage and improved salinity tolerance (Hasegawa
et al, 2000; Mittova et al, 2003; Demiral and Turkan,
2005; Hoque et al, 2007a, b, 2008; Banu et al, 2009;
El-Shabrawi et al, 2010). In order to evaluate the
relationship between salinity tolerance and antioxidant
response, the activity of ROS-scavenging antioxidant
enzymes was measured. The activity of antioxidant
enzymes differed significantly in response to salinity
stress. In this study, CAT and APX activities increased
with increased salt concentrations in salt-tolerant rice
genotypes but the activities in the salt-sensitive
genotype (BRRI dhan 28) decreased with increased
salinity, as compared to the control (Fig. 1). Previous
reports support that increased salinity level decreases
the activity of antioxidant enzymes including CAT in
salt-sensitive rice, but increases CAT activity in
salt-tolerant rice (Demiral and Turkan, 2005;
El-Shabrawi et al, 2010; Hasanuzzaman et al, 2014).
POX activity was considerably decreased in response
to increased salinity in all the genotypes irrespective
of their tolerance levels (Fig. 1). Dogan et al (2011)
also showed that high salinity significantly decreases
APX activity in salt-sensitive soybean. Our results are
consistent with the observations of many researchers
who reported that APX activity coordinated with CAT
and POX activities confers salt tolerance in rice as
well as in cotton (Meloni et al, 2003; Vaidyanathan et al,
Mohammad Golam KIBRIA, et al. Antioxidant Defense Mechanisms of Salinity Tolerance in Rice
2003; Demirel and Turkan, 2005).
Effects of salinity on K+/Na+ ratio in rice shoots
and roots
A high cytosolic K+/Na+ ratio is essential for reducing
salinity-induced oxidative damage. Enhanced influx of
Na+ and inhibition of K+ uptake by plants under salt
stress caused disturbances in K+/Na+ homeostasis,
leading to toxic effects on plants (Zhu, 2003). The
K+/Na+ ratio in both shoots and roots was significantly
higher in the salt-tolerant genotypes than in the
salt-sensitive one (Fig. 1). In the present study,
differences in K+/Na+ ratio between rice genotypes
suggest that Na+ exclusion from leaf tissues appears to
play an important role in the salt tolerance of rice by
maintaining the optimal K+/Na+ ratio. High salt (Na+)
uptake competes with the uptake of other nutrient ions,
especially K+, leading to K+ deficiency. It is often
found that many glycophytes exhibiting enhanced
tolerance to salinity stress have higher ability for
sodium exclusion, maintaining high levels of K+/Na+
ratio (Zhu, 2001).
CONCLUSIONS
It is clear that increased antioxidant enzyme (CAT and
APX) activity, chlorophyll content, proline content
and K+/Na+ ratio in rice could be some of the
physiological and biochemical mechanisms underlying
salinity tolerance in plants. It is evident that no single
parameter could be responsible for salinity tolerance
in any of the studied rice genotypes whereas a
combination of different characteristics contributes to
the salinity tolerance. These findings may serve as in
vitro selection criteria for salt tolerance in rice. On the
basis of growth performance and biochemical assays,
BINA dhan 10 showed a higher tolerance to salinity
stress than the other genotypes. Therefore, further
research should be focused on bio-molecular
mechanisms involved in salinity stress tolerance for
the determination of key pathways controlling salinity
tolerance in plants.
ACKNOWLEDGEMENTS
This study was supported by a grant from the Ministry
of Education, Government of Bangladesh. The authors
are grateful to Gretchen Elizabeth DYKES, PhD,
Department of Plant and Soil Sciences, University of
161
Delaware, USA, for English editing of this article.
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