J.

Agronomy & Crop Science (2008) ISSN 0931-2250

CHILLING/FREEZING STRESS

Chilling Tolerance in Hybrid Maize Induced by Seed Priming

with Salicylic Acid
M. Farooq1, T. Aziz1, S. M. A. Basra2, M. A. Cheema1 & H. Rehman2
1 Department of Agronomy, University of Agriculture, Faisalabad, Pakistan
2 Department of Crop Physiology, University of Agriculture, Faisalabad, Pakistan

Keywords Abstract
antioxidants; chilling stress; emergence;
maize; salicylic acid; seed priming; seedling The optimum temperature for maize germination is between 25 and 28 C.
growth Poor and erratic germination at suboptimal temperature is the most important
hindrance in its early sowing. This study was conducted to induce chilling tol-
Correspondence erance in hybrid maize (Zea mays L.) by seed priming with salicylic acid (SA)
Dr M. Farooq
and to unravel the background biochemical basis. For seed priming, maize
Department of Agronomy, University of
Agriculture, Faisalabad 38040, Pakistan
hybrid (Hycorn 8288) seeds were soaked in 50, 100 and 150 ppm (mg l)1) aer-
Tel.: +92 41 9200161-9/2936 ated solutions of SA for 24 h and were dried back. Treated and untreated seeds
Fax: +92 41 9201098 were sown at 27 C (optimal temperature) and at 15 C (chilling stress) under
Email: farooqcp@gmail.com controlled conditions. Performance of maize seedlings was hampered under
chilling stress. But seed priming with SA improved the seedling emergence,
Accepted January 23, 2008 root and shoot length, seedling fresh and dry weights, and leaf and root score
considerably compared with control both at optimal and chilling temperatures.
doi:10.1111/j.1439-037X.2008.00300.x
However, priming in 50 mg l)1 SA solution was more effective, followed by
priming in 100 mg l)1 SA solution. Seed priming with SA improved the chill-
ing tolerance in hybrid maize mainly by the activation of antioxidants (includ-
ing catalase, superoxide dismutase and ascorbate peroxidase). Moreover,
maintenance of high tissue water contents and reduced membrane permeability
also contributed towards chilling tolerance.

gens is created by the induction of pathogenesis-related

Introduction
Optimum temperature for maize (Zea mays L.) germina-
tion is between 25 and 28 C. Poor and erratic germina-
tion at suboptimal temperature is the most important
hindrance in its early sowing (Stewart et al. 1990, Greaves
1996). Moreover, chilling causes shoot water deficit in
maize crop because of its thermophilic nature. This water
deficit is caused by a reduction in the root water uptake
greater than in the leaf transpiration rate during chilling
(Stewart et al. 1990).
Salicylic acid (SA) is a phytohormone of phenolic nat-
ure. It is ubiquitous in plants generating a considerable
impact on plant growth, development, mineral uptake
and transport, photosynthesis, and transpiration. It also
induces specific changes in leaf anatomy and chloroplast
structure. SA is recognized as an endogenous signal,
mediating in plant defense system in particular against
pathogens (Hayat and Ahmad 2007). Resistance to patho-
proteins (Davies 2004).
Numerous studies reported that SA can improve plant
resistance to various abiotic stresses including ultraviolet
light, drought, salt and temperature extremes (Yalpani
et al. 1994, Dat et al. 1998, Mishra and Choudhuri 1999,
Senaratna et al. 2000, He et al. 2005, Ogawa et al. 2005,
Horváth et al. 2007). Recently, Gunes et al. (2007) pro-
posed that SA acts as an endogenous signal molecule
responsible for inducing tolerance against many abiotic
stresses in plants. SA could provide some protection for
cell structure of chilling-stressed banana (Musa paradisia-
cal L.) seedling (Kang et al. 2007).
Salicylic acid analogue, 4-hydroxybenzoic acid improved
the drought tolerance of the winter wheat (Triticum aes-
tivum L.) cv. Cheyenne and the freezing tolerance of the
spring wheat cv. Chinese Spring (Horváth et al. 2007). In
another study, exogenous SA treatment decreased relative
electrolyte leakage in grape (Vitis vinifera L.) leaves under

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Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 161–168 161
Farooq et al.
heat or cold stress, indicating that SA can induce intrinsic
heat or cold tolerance in grape by maintenance of
relatively higher activities of ascorbate peroxidase (APX),
glutathione reductase, monodehydroascorbate and redox
ratio in the ascorbate–glutathione pool under normal tem-
perature and under heat or cold stress (Wang and
Li 2006). He et al. (2005) also reported that SA application
concealed the reactive oxygen species (ROS) production
and enhanced the activities of superoxide dismutase
(SOD) and catalase (CAT) activities, which induced heat
tolerance in Kentucky bluegrass.
Antioxidants have been found to play a vital role in
improving chilling tolerance (Foyer and Fletcher 2001,
Blokhina et al. 2003). For instance, Prasad (1997)
reported that CAT plays an important role in acclimation
to chilling temperature in maize seedlings. Sundar et al.
(2004) suggested that increase in several components of
the antioxidant system in cold-treated guayule (Parthe-
nium argentatum) plants is responsible to mitigate the
oxidative damages.
Seed enhancements (including seed priming with or
without plant growth regulator) may improve the germina-
tion and seedling establishment at suboptimal temperatures.
Several studies report the benefits of priming on germina-
tion to improve the germination and stand establishment
(Basra et al. 2005, Farooq et al. 2006a, Farooq et al. 2006b,
Farooq et al. 2006c). Moreover, priming has also been
found effective to increase the germination under chilling
conditions. For example, seed priming with CaCl2 and gly-
cinebetaine has been found to improve chilling tolerance in
wheat (Farooq et al. 2008a) and hybrid maize (Farooq et al.
2008b) respectively. In another study, Basra et al. (2006)
found that seed priming with SA improve the germination
and early seedling growth in coarse and fine rice.
Until now, no study has been conducted to investigate
the possibility of chilling tolerance in maize by seed prim-
ing with SA. This study therefore was carried out with
the objective to induce chilling tolerance in maize by seed
priming with SA, and to explore the possible biochemical
basis of chilling tolerance.
Materials and Methods
Seeds of maize hybrid (Hycorn 8288) obtained from ICI
Life Sciences, Sahiwal, Pakistan having initial seed mois-
ture contents 8.54 % (on dry weight basis) were used in
the study. For all treatments, selected healthy seeds were
used in same numbers.
Salicylic acid seed treatments
For seed priming, seeds were soaked in 50, 100 and
150 ppm (mg l)1; w/v) aerated SA solution for 24 h at
162
28 ± 2 C. The ratio of seed weight to solution volume
was 1 : 5 (g ml)1) (Farooq et al. 2006a). After each treat-
ment, seeds were given three surface washings with dis-
tilled water and dried back closer to original moisture
level under forced air at 27 C ± 3, sealed in polythene
bags and stored in a refrigerator at 5 C until use (Lee
and Kim 2000). Untreated dry seeds were taken as
control.
Seed germination and seedling vigour evaluation
Treated and untreated seeds were sown in 10 kg plastic
pots (15 in each) containing moist acid/water washed
sand and placed in a growth room with a photosyntheti-
cally active photon flux density of 350 mmol m)2 s)1 and
a photoperiod of 14/10 h light/dark. Experimental design
was completely randomized with five replications. One set
of pots was placed at 27 C (optimal temperature, con-
trol), while the other was kept at 15 C (chilling stress)
during the whole period of study. From each pot, number
of emerged seedlings was recorded daily according to the
seedling evaluation handbook of Association of Official
Seed Analysis (1990). Time taken to 50 % emergence of
seedlings (E50) was calculated according to the following
formulae of Coolbear et al. (1984) modified by Farooq
et al. (2005):
½ðN=2Þ  ni ðtj  ti Þ
E50 ¼ ti þ
nj  ni
where N is the final number of emerged seeds, and ni and
nj the cumulative number of seeds emerged by adjacent
counts at times ti and tj when ni < N/2 < nj.
Mean emergence time (MET) was calculated according
to the equation of Ellis and Roberts (1981) as under:
P
Dn
MET ¼ P
n
where n is the number of seeds, which were emerged on
day D, and D is the number of days counted from the
beginning of emergence.
Energy of emergence (EE) was determined on fourth
day of seed sowing; it is the number of emerged seedlings
expressed in percentage on fourth day after sowing
(Farooq et al. 2006b). While, coefficient of uniformity
of emergence (CUE) was calculated using the following
formulae of Bewley and Black (1985):
P
n
CUE ¼ P  
ðt  tÞ2 n
where t is the time in days, starting from day 0, the day
of sowing, and n is the number of seeds completing
emergence on day t and t is equal to MET.
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Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 161–168

On 15th day after emergence, samples (second top leaf
from main tiller) for biochemical analysis were collected
from control and chilling stressed plants. At the same
day, seedlings were tested for vigour after carefully
removing from the sand. Number of leaves, secondary
roots, and seedling shoot and root length were recorded
of 10 seedlings per replicate and averaged. Number of
leaves and secondary roots per seedling were designated
as leaf and root score respectively. Seedling fresh weight
was determined immediately after harvest while dry
weight was taken after drying at 70 C for 7 days.
Membrane permeability
Membrane permeability was estimated by measuring the
electrolyte leakage according to the method of Blum and
Ebercon (1981). Six leaf samples were rinsed with distilled
water and immersed in 6 ml of distilled water for 12 h.
The conductivity of the solution (C1) was measured with
a conductivity meter (Model DDS-11A; Shanghai Leici
Instrument Inc., Shanghai, China). Samples were then
heated in boiling water for 20 min and then cooled to
room temperature. The conductivity of killed tissues (C2)
was again measured. Membrane permeability was calcu-
lated as the ratio between C1 and C2.
Relative water content
To estimate relative water contents (RWC), weighed
amount of fresh leaves (0.5 g; Wf) were rinsed and put in
a tube containing water until the weight of the leaves was
constant. The saturated leaves were weighed (Ws) and
then dried for 24 h at 80 C for determinations of the
dry weigh (Wd). RWC was calculated by the following
formula:
Wf  Wd
RWC ¼  100%
Ws  Wd
Extraction and determination of enzyme activities
For a-amylase activity, 1 g ground leaf sample was mixed
with 10 ml phosphate buffer (pH 7.0) and left for 24 h at
4 C. The enzyme activity was determined from the
supernatant by the di-nitrosalicylic acid (DNS) method
(Bernfeld 1955).
Total extractable SOD activity was measured following
the protocol described by McCord and Fridovitch (1969).
The inhibition of the colour formation (measured at
560 nm) was determined by addition of 0–50 ll of the
extract to a reaction mixture containing 50 mm HEPES
(4-(2-hydroxyethyl-1-piperazineethanesulfonic acid)/KOH
buffer (pH 7.8), 0.05 units xanthine oxidase, 0.5 mm
ª 2008 The Authors
Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 161–168
Chilling Tolerance in Hybrid Maize
nitroblue tetrazolium and 4 mm xanthine. One unit of
SOD activity is equivalent to the volume of extract
needed to cause 50 % inhibition of the colour reaction.
Catalase activity was determined according to modified
method of Luck (1974). Enzyme extract (50 ll) was
added to 3 ml of hydrogen-peroxide-phosphate buffer
(pH 7.0). The time required for the decrease in the absor-
bance from 0.45 to 0.40 was noted. Enzyme solution con-
taining hydrogen peroxide-free phosphate buffer was used
as control.
Ascorbate peroxidase activity was determined following
the method of Nakano and Asada (1987) with slight
modification. Ascorbate oxidation to dehydroascorbate
was followed at 265 nm in 1 ml reaction mixture contain-
ing 50 mm HEPES/KOH (pH 7.6), 0.1 mm EDTA,
0.05 mm ascorbate, 10 ll extract and 0.1 mm H2O2.
Extraction and estimation of soluble sugars
For soluble sugars, 1 g ground leaf sample was mixed
with 10 ml distilled water and left for 24 h at 25 C
(Lee and Kim 2000). The mixture was filtered (with
Whatman No. 42) and the final volume made to 10 ml
with distilled water. Total soluble sugars were determined
by the phenol sulphuric method (Dubois et al. 1956).
Statistical analysis
The experiments were conducted twice; data were pooled
and subjected to statistical analysis using costat com-
puter package (CoHort Software, Berkeley, CA, USA).
Graphical presentation of data was carried out using
Microsoft Excel program. For comparison of treatment
means, standard errors were computed using Microsoft
Excel program. Parallels were drawn between antioxidants
and electrolyte leakage and RWC.
Results
Although chilling considerably delayed the seedling emer-
gence, and decreased the synchronization and final emer-
gence than at optimal temperature; seed priming with SA
improved the emergence time and uniformity at both the
temperatures (Figs 1–3). Under both normal and stress
conditions, seed priming with 50 mg l)1 SA was the best
to reduce the time to 50 % emergence (E50) and MET
and increase the EE and CUE compared with other treat-
ments (Fig. 1–3). Although, statistically maximum final
emergence percentage (FEP) under optimal temperature
was recorded from seed treatment with 50 mg l)1 SA;
however, at low temperature statistically maximum FEP
was recorded from the seed treated with 100 mg l)1 SA
(Fig. 2b). Likewise, seed treatment with 150 mg l)1 SA
163
Farooq et al.

(a) 5 Optimum temp. 1 Optimum temp.

Low temp. Low temp.
4 0.8
E50 (days)

3 0.6

CUE
2 0.4

1 0.2

0 0
Control 50 ppm 100 ppm 150 ppm Control 50 ppm 100 ppm 150 ppm
Pre-sowing salicylic acid seed treatments
(b) 8 Optimum temp.
Low temp. Fig. 3 Influence pre-sowing salicylic acid seed treatments on the on
6 the coefficient of uniformity of emergence in maize at optimal and
MET (days)

low temperatures ± S.E.

4

2 (a) 30 Optimum temp.

Low temp.
25

Shoot length (cm)

0
Control 50 ppm 100 ppm 150 ppm 20
Pre-sowing salicylic acid seed treatments 15
10
Fig. 1 Influence of pre-sowing salicylic acid seed treatments on the
(a) time to 50 % emergence and (b) mean emergence time in maize 5
at optimal and low temperatures ± S.E. 0
Control 50 ppm 100 ppm 150 ppm

(b) 30 Optimum temp.

Optimum temp. Low temp.
Root length (cm)

Low temp. 25
(a) 80
20

60 15
10
EE (%)

40 5
0
Control 50 ppm 100 ppm 150 ppm
20
Pre-sowing salicylic acid seed treatments

0
Control 50 ppm 100 ppm 150 ppm Fig. 4 Influence of pre-sowing salicylic acid seed treatments on the
(a) shoot and (b) root length in maize at optimal and low tempera-
(b)
100 Optimum temp. tures ± S.E.
Low temp.

80 of SA could not improve these attributes. However, both

50 and 100 mg l)1 SA treatments improved the root score
FEP (%)

60
40
20
0 was no difference amongst the SA treatments each at
Control 50 ppm 100 ppm
Pre-sowing salicylic acid seed treatments
Fig. 2 Influence of pre-sowing salicylic acid seed treatments on the
(a) energy of emergence and (b) final emergence percentage in maize
at optimal and low temperatures ± S.E.
increased the E50 at optimal temperature compared with
control (Fig. 1a).
Under both optimal and stress conditions, 50 mg l)1
SA appreciably improved the shoot and root length
(Fig. 4), seedling fresh and dry weight (Fig. 5) and leaf
score (Fig. 6a) compared with control, while other levels
compared with control (Fig. 6b).
All the seed priming treatments improved the RWC
under both optimal and stress conditions. However, there
150 ppm normal and low temperature (Fig. 7a). Similarly, all the
SA treatments decreased the electrolyte leakage in maize
leaves at both normal and stress conditions; however,
minimum leakage was recorded from 50 ppm SA treat-
ment under both conditions (Fig. 7b).
At both temperatures, all SA treatments improved the
amylase activity (Fig. 8a) and soluble sugars (Fig. 8b)
compared with control. However, maximum sugars, and
amylases were recorded from 50 ppm SA treatment under
both conditions (Fig. 8).
At normal and low temperatures, seed priming with SA
improved the SOD (Fig. 9a), CAT (Fig. 9b) and APX

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164 Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 161–168
 Chilling Tolerance in Hybrid Maize

(a) (a) 90 Optimum temp.

Low temp.
180 Optimum temp.
Seedling fresh weight (mg)

Low temp.
75
60

RWC (%)
135
45
90
30

45 15
0
0 Control 50 ppm 100 ppm 150 ppm
Control 50 ppm 100 ppm 150 ppm
(b)
(b) 30 Optimum temp.

Electrolyte leakage (%)

45 Optimum temp. Low temp.
Low temp. 25
Seedling dry weight (mg)

36 20
15
27
10
18 5
0
9 Control 50 ppm 100 ppm 150 ppm
Pre-sowing salicylic acid seed treatments
0
Control 50 ppm 100 ppm 150 ppm
Fig. 7 Influence of pre-sowing salicylic acid seed treatments on the
Pre-sowing salicylic acid seed treatments
(a) relative water content (RWC) and (b) electrolyte leakage in maize
Fig. 5 Influence of pre-sowing salicylic acid seed treatments on the at optimal and low temperatures ± S.E.
(a) seedling fresh weight and (b) seedling dry weight in maize at opti-
mal and low temperatures ± S.E.
(a) 12 Optimum temp.
Low temp.
α-amylase activity (unit)*

9
(a) 4 Optimum temp.
Low temp.

3 6
Leaf score

2 3

1
0
Control 50 ppm 100 ppm 150 ppm
0 (b)
Control 50 ppm 100 ppm 150 ppm
Soluble sugars (mg g–1 fresh weight)

25 Optimum temp.
Low temp.
(b) Optimum temp.
Low temp. 20
8

6 15
Root score

4
10
2
5
0
Control 50 ppm 100 ppm 150 ppm 0
Pre-sowing salicylic acid seed treatments Control 50 ppm 100 ppm 150 ppm

Fig. 6 Pre-sowing salicylic acid seed treatments on the (a) leaf score
and (b) root score in maize at optimal and low temperatures ± S.E.
(Fig. 10) compared with control. However, the maximum
SOD and CAT were recorded from 50 ppm SA treatment
under both conditions (Figs 8 and 9). There was no dif-
ference in APX activity amongst all the SA treatments at
optimal temperature; however, at low temperature similar
Fig. 8 Influence of pre-sowing salicylic acid seed treatments on the
(a) a-amylase activity and (b) soluble sugars in maize at optimal and
low temperatures ± S.E. *One unit of the enzymes activity is the
amount of enzyme which released 1 lmol of maltose by 1 mL original
enzyme solution in 1 min.
APX activity was recorded in 50 and 150 ppm SA treat-
ments. But APX activity was less at 100 ppm than other
SA treatments (Fig. 10).

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Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 161–168 165
Farooq et al.

SOD (Unit g–1 protein)* (a) Optimum temp. was negative correlations between antioxidants and elec-
Low temp.
50 trolyte leakage at optimal and chilling temperatures
40 (Table 1).
30
20
Discussion
10
0 This study exposed that SA seed treatments can consider-
Control 50 ppm 100 ppm 150 ppm ably improve the low temperature tolerance in hybrid
(b)
maize. Although under chilling, emergence, early seedling
30 Optimum temp.
CAT (µmol min–1 g–1 protein)

Low temp. growth and metabolism were disturbed, SA treatments

25
improved the performance under both the optimal and
20 low temperatures. Under both normal and stress condi-
15 tions, seed priming with 50 ppm proved the best treat-
10 ment to improve the emergence, early seedling growth,
5
antioxidant activities and general metabolism in hybrid
maize. SA levels higher than 50 mg l)1 were not effective
0
Control 50 ppm 100 ppm 150 ppm rather reduced the root and shoot length (Fig. 3), seedling
Pre-sowing salicylic acid seed treatments fresh and dry weights (Fig. 5), leaf and root score (Fig. 6)
and increased the electrolyte leakage (Fig. 7b) compared
Fig. 9 Influence of pre-sowing salicylic acid seed treatments on the
with control. Higher SA concentration has been found to
(a) superoxide dismutase (SOD) and (b) catalase (CAT) in maize at
be toxic because of its possible involvement in reduction
optimal and low temperatures ± S.E. *One unit of SOD activity is
equivalent to the volume of extract needed to cause 50% inhibition in cell division and negative effects on photosynthetic
of the colour reaction. machinery (Elmer 2005). Improved root and shoot
length, and seedling fresh and dry weights might be due
Optimum temp.
Low temp.
to increased cell division within the apical meristem,
APX (µmol min–1 g–1 protein)

15
which caused an increase in plant growth (Huang and
12 Villanueva 1992). SA regulates cell growth by specifically
9
affecting cell enlargement, endoreduplication and/or cell
division (Vanacker et al. 2001), protecting the cell struc-
6 ture (Kang et al. 2007).
3 By chilling electrolyte leakage of maize leaves consider-
ably increased. However, all the SA seed treatments
0
Control 50 ppm 100 ppm 150 ppm reduced the electrolyte leakage. Enhanced electrolyte leak-
Pre-sowing salicylic acid seed treatments age was considered to be a symptom of stress-induced
membrane damage and deterioration (Feng et al. 2003).
Fig. 10 Influence of pre-sowing salicylic acid seed treatments on the
ascorbate peroxidase (APX) in maize at optimal and low tempera-
Under low temperature stress, RWC were also decreased.
tures ± S.E. Maize plants exposed to chilling stress often show water-
stress symptoms because of chilling-induced water stress
Table 1 Correlations coefficients (r) of antioxidants with changes in (Melkonian et al. 2004). Nonetheless, SA treatments
membrane electrolyte leakage and relative leaf water content of improved the tissue water status most likely through
maize under normal and chilling stress conditions osmotic adjustment and/or changes in cell wall elasticity
Electrolyte leakage Relative water contents (Bandurska and Stroi ski 2007; Korkmaz et al. 2007).
Exogenously applied SA decreased the electrolyte leakage
Antioxidants Optimal Chilling stress Optimal Chilling stress under heat or cold stress (Wang and Li 2006), which
SOD )0.92** 0.98*** 0.82* 0.97*** indicates that SA can induce intrinsic heat or cold toler-
CAT )0.96** )0.99*** 0.81* 0.95*** ance by maintenance of relatively higher activities of anti-
APX )0.98*** )0.99*** 0.93** 0.93** oxidants (Wang and Li 2006) as is evident from strong
correlation between antioxidants and RWC (Table 1).
*P < 0.05, **P < 0.01, ***P < 0.001, (n = 4). SOD, superoxide
dismutase; CAT, catalase; APX, ascorbate peroxidase.
If scavenging system against ROS is not working effi-
ciently, they may change the plant metabolism principally
Parallels drawn between antioxidants and electrolyte by structural modification of proteins to make them more
leakage and RWC indicated that strong positive corre- susceptible to proteolytic degradation (Pell and Dann
lations between antioxidants and RWC. While, there 1991). ROS in plants are removed by a variety of antioxi-

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166 Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 161–168

dant enzymes and/or lipid- and water-soluble scavenging
molecules (Foyer et al. 1994), the antioxidant enzymes
are the most effective against oxidative damage (Halliwell
and Gutteridge 1999). Enzymes of SOD, CAT and APX
are the most important in this regard, SOD catalyses the
dismutation of O2) to H2O2 and O2, while CAT and APX
scavenge H2O2 (Foyer et al. 1994). The present results
show that enzymatic antioxidant activities of maize seed-
lings were improved upon exposure to low temperature
stress compared with that of maize seedlings grown at
normal temperature. This indicates that maize hybrid
Hycorn 8288 has the ability to activate its antioxidant sys-
tem upon exposure to stress. However, the treatment of
seeds with SA increased the SOD, CAT and APX activities
under both growth conditions. Higher CAT activity in
leaves under chilling stress suggests a more efficient scav-
enging system which may result in better protection
against ROS during stress. In comparison with the pres-
ent study, Feierabend et al. (1992) reported reduced CAT
activity in plants exposed to low temperature and bright
light because of photo-inactivation of CAT. But Prasad
(1997) concluded that exposure to low temperatures
causes an increase in CAT activity, which supports our
results. This increased antioxidant production reduced
the ROS-based damages in the plant system as evident
from strong correlation between membrane electrolyte
leakage and antioxidant system (Table 1). However, these
results are in contrast to Dat et al. (2007) who reported
that SA acts as potent inhibitor of haeme-containing anti-
oxidant enzymes such as CAT and APX and thus stimu-
late the ROS accumulation during various stresses. But to
us, this happens only during programmed cell death not
always (Holuigue et al. 2007).
In conclusion, seed priming with SA at low concentra-
tion can improve the low temperature tolerance in hybrid
maize by the activation of antioxidant system and mainte-
nance of high tissue water contents. Performance of
hybrid maize was also improved at normal temperature.
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ª 2008 The Authors
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