J.

Agronomy & Crop Science (2008) ISSN 0931-2250

CHILLING/FREEZING STRESS

Activation of Antioxidant System by KCl Improves the

Chilling Tolerance in Hybrid Maize
M. Farooq, T. Aziz, Z. A. Cheema, M. Hussain & A. Khaliq
Department of Agronomy, University of Agriculture, Faisalabad, Pakistan

Keywords
antioxidants; chilling stress; emergence;
maize; ROS; tolerance
Correspondence
Dr Muhammad Farooq
Department of Agronomy
University of Agriculture
Faisalabad 38040, Pakistan
Tel.: +92 41 9200161-9/2917
Fax: +92 41 9200605; +92 41 9201098
Email: farooqcp@gmail.com
Accepted July 24, 2008
doi:10.1111/j.1439-037X.2008.00334.x
Abstract
As maize is a chilling-sensitive crop, low temperatures during the early stages
of development can be injurious to crop growth and development. Prime
mechanism behind chilling-induced damage is oxidative stress. This study was
undertaken to improve the chilling tolerance in hybrid maize by seed priming
with KCl. For priming, seeds of the maize hybrid Hycorn 8288 were soaked in
50, 100 and 150 mg l)1 aerated solution of KCl for 24 h and then re-dried
close to original weight. Primed and untreated seeds were sown at 27 C (opti-
mal temperature) and at 15 C (chilling stress) under controlled conditions.
Seed priming improved the performance of maize under both normal and
stress conditions. It was found that the chilling tolerance in maize is well asso-
ciated with the enhanced capacity of the anti-oxidative system. Priming with
KCl significantly improved the chilling tolerance mainly by the activation of
antioxidants including catalase, superoxide dismutase and ascorbate peroxidase
enzymes. KCl treatments also improved the germination rate and time, root
and shoot length, and fresh and dry weights of seedlings compared with con-
trol. Soluble sugars and a-amylase activity determined as general metabolic
indicators of stress were also improved by seed priming with KCl. Other possi-
ble bases of chilling tolerance in maize included maintenance of high tissue
water contents, reduced electrolyte leakage and carbohydrate metabolism. Seed
treatment with 100 mg l)1 KCl was the best treatment to improve the perfor-
mance of hybrid maize both under normal and chilling stress conditions.

plants (Allen and Ort 2001). ROS levels in plants also

Introduction
Plants are subjected to various kinds of biotic and abiotic
stresses including drought, heat, salinity and chilling,
which hamper the seedling establishment, allometry and
economic yield (Afzal et al. 2006, Wahid 2007, Farooq
et al. 2008a,b,c,d,e,f). Chilling stress alters the fluidity of
membrane lipids and thus disturbs the membrane struc-
ture (Marschner 1995). Absorbed radiation energy from
sunlight surpasses the capacity of chloroplasts to use it in
CO2 fixation, and the glut energy is alternatively used to
convert O2 to reactive oxygen species (ROS) under chill-
ing stress (Huner et al. 1998, Foyer et al. 2002, Apel and
Hirt 2004). Photosynthetic electron transport, stomatal
conductance, rubisco activity and CO2 fixation are the
major targets impaired by low-temperature stress in
increase because of perturbations of chloroplastic and
mitochondrial metabolism (Van Camp et al. 1998,
Kratsch and Wise 2000, Foyer and Fletcher 2001, Neill
et al. 2002).
The ability to adjust antioxidant systems to changing
ROS concentrations may be vital to all species under
conditions of stress (Kocsy et al. 1997, 2001, Foyer et al.
2002). Hence, plants have developed active oxygen-
scavenging systems consisting of several antioxidant
enzymes, such as superoxide dismutase (SOD), ascorbate-
peroxidase (APX), glutathione reductase (GR) and
catalase (CAT), and some low molecules of non-enzyme
antioxidants such as ascorbic acid (AA) and reduced
glutathione (GSH) (Bowler et al. 1992, Halliwell
and Gutteridge 1999, Hasegawa et al. 2000, Fazeli et al.

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438 Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 438–448

2007, Farooq et al. 2008e). These systems protect mem-
branes from the deleterious effects of ROS such as super-
oxide radicals, hydrogen peroxide (H2O2), hydroxyl
radicals and singlet oxygen (Bowler et al. 1992, Noctor
and Foyer 1998; Farooq et al. 2008a). For example, super-
oxide radical is dismutated to H2O2 by SOD, and CAT
and APX metabolize H2O2 into H2O.
Seed priming is a technique by which seeds are par-
tially hydrated to a point where germination processes
begin but radicle emergence does not occur (Bradford
1986, Farooq et al. 2005, 2006a,b,c). Various seed-priming
techniques have been reported to improve the germina-
tion and seedling establishment at low temperatures. In
rice, seed priming with various concentrations of proline,
betaine, putrescine, spermidine and spermine increased
low temperature tolerance (Naidu and Williams 2004).
Sasaki et al. (2005) noted that growth was promoted by
H2O2 treatment under low temperature in a greenhouse.
In another study, Zheng et al. (2002) also reported signif-
icantly improved chilling tolerance in rice. While in
maize, priming with salicylic acid, glycinebetaine and
CaCl2 has been found effective to improve the chilling
tolerance (Farooq et al. 2008b,c,d).
Growing evidences advocate that the improvement of
potassium (K) nutritional status of plants can greatly
lower the ROS production by reducing the activity of
NAD(P)H oxidases and maintaining photosynthetic elec-
tron transport (Cakmak 2005). In another study, it was
found that increasing K concentration in irrigation water
provided important protection against stem damage from
low temperatures during night (Kafkafi 1990). Priming
with KCl has been found to be the most effective tech-
nique in improving the growth of rice nursery seedling
(Farooq et al. 2007) and stand establishment in coarse
rice (Farooq et al. 2006a,b,c).
In maize, even temperatures below 15 C may induce
chilling stress (Pál and Nagy 2002, Holá et al. 2003).
When maize is sown during the month of February in
Pakistan, mean daily temperature is lower than
15 ± 2 C, which is quite lower than that required for
optimum maize cultivation. This warrants finding strate-
gies to accomplish optimal stand and seed priming with
KCl may be a pragmatic approach.
No information is available reporting the leeway to
induce chilling tolerance in hybrid maize by KCl seed
treatments. It is hypothesized that maize seed priming
with KCl solution may alleviate the oxidative damage
with the enhanced activities of enzyme antioxidants and
promote seedling emergence and subsequent growth.
Therefore, this study was carried out to explore the
role of KCl seed treatments in improved chilling
tolerance in hybrid maize by the activation of anti-
oxidants.
ª 2008 The Authors
Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 438–448
Chilling Tolerance in Maize
Materials and methods
Source of seed
Seeds of maize hybrid (Hycorn 8288) obtained from
ICI Life Sciences, Sahiwal, Pakistan (8.54 % seed
moisture content), were used in the study. For all
treatments, the same numbers of selected healthy seeds
were used.
Seed priming treatments
For priming, maize seeds were soaked in an aerated solu-
tion of KCl of concentrations 50, 100 and 150 mg l)1 for
24 h at 28 ± 2 C. Seed weight to solution volume ratio
was 1:5 (w/v) (Farooq et al. 2006a). After each treatment,
seeds were rinsed thoroughly with distilled water and
dried closer to the original moisture level under forced
air at 27 ± 3 C, 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.
Emergence and seedling vigour evaluation
Treated and control 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 15-h light period, 30/
24 C day/night temperatures and 70 % relative humidity.
The number of emerged seeds was recorded daily accord-
ing to the seedling evaluation as mentioned in the hand-
book of the Association of Official Seed Analysts (1990).
Time taken for 50 % emergence of seedlings (E50) was
calculated according to the formulae of Coolbear et al.
(1984) modified by Farooq et al. (2005). Mean emergence
time (MET) was calculated using the formulae of Ellis and
Roberts (1981). Energy of emergence (EE) was determined
on the fourth day of seed sowing (Farooq et al. 2006b).
Coefficient of uniformity of emergence (CUE) was calcu-
lated using the formulae of Bewley and Black (1985). On
day 15 after sowing, the seedlings were tested for vigour
after carefully removing from the sand. The numbers of
secondary roots and leaves were taken as root and leaf
score, respectively. Seedling fresh weight was determined
immediately after harvest while dry weight was taken after
drying at 70 C for 7 days.
Membrane stability
Membrane stability was determined in terms of electrolyte
leakage according to the method of Blum and Ebercon
(1981). Six leaf segments of about equal size were rinsed
with distilled water and immersed in 6 ml of distilled
water for 12 h followed by the measurement of electrical
conductivity (EC1) of the solution with a conductivity
439
Farooq et al.
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 (EC2) was again
measured. Membrane stability was calculated as the ratio
between EC1 and EC2.
Relative water contents
Fresh leaves (0.5 g; Wf) were washed in water until con-
stant weight of the leaves was attained. The saturated
leaves were weighed (WS) and then dried for 24 h at
80 C for determinations of dry weight (Wd). Relative
water content (RWC) was calculated by the following
formula:
RWC ¼ ðWf  Wd Þ=ðWS  Wd Þ  100%:
Sugar metabolism
To determine the a-amylase activity, 1 g of ground leaf
sample was mixed with 10 ml of phosphate buffer
(pH 7.0) and left for 24 h at 4 C. The enzyme activity
was determined from the supernatant by the DNS
method (Bernfeld 1955). To determine total soluble sug-
ars, 1 g of ground leaf sample was mixed with 10 ml of
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 was made to 10 ml with distilled
water. Total soluble sugars were determined by the phe-
nol sulphuric acid method (Dubois et al. 1956).
Antioxidant activities
Total extractable SOD activity was determined following
the method of McCord and Fridovitch (1969). Inhibition
of colour formation (measured at 560 nm) was
determined by the addition of 0–50 ll of the extract to a
reaction mixture containing 50 mm HEPES/KOH buffer
(pH 7.8), 0.05 units xanthine oxidase, 0.5 mm nitroblue
tetrazolium and 4 mm xanthine. One unit of SOD activity
equalled the volume of extract needed to cause 50 %
inhibition of the colour reaction. Catalase (CAT) activity
was measured following the 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 absorbance from 0.45 to
0.40 was noted. Enzyme solution containing H2O2-free
phosphate buffer was used as control. Enzyme activity
was expressed in mmol of H2O2 consumed
min)1 mg)1 chl. Ascorbate peroxidase (APX) activity was
estimated according to the method of Nakano and Asada
(1987) with slight modification. Ascorbate oxidation to
440
dehydroascorbate (DHA) was followed at 265 nm in a
1-ml reaction mixture containing 50 mm HEPES/KOH
(pH 7.6), 0.1 mm EDTA, 0.05 mm ascorbate, 10 ll extract
and 0.1 mm H2O2.
Statistical analysis
The experiment was laid out in completely randomized
design in factorial arrangement with four replications.
The experiments were conducted twice; data were pooled,
and subjected to statistical analysis using costat com-
puter package (CoHort Software, Berkeley, CA, USA).
Least significant difference (LSD) test was applied to
compare the treatment mean values. Graphical presenta-
tion of data was carried out by using the Microsoft Excel
program (Microsoft Corporation, Los Angeles, CA, USA).
For comparison of treatment mean values, standard
errors were computed using Microsoft Excel program.
Correlation coefficients were computed in order to
assess the relationships of various parameters for seed-
ling growth with general metabolites and antioxidant
activity.
Results
Germination, early seedling growth, membrane stability,
RWC, starch metabolism, antioxidant activities were sig-
nificantly affected (P £ 0.05) by chilling stress (Figs 1–9).
Chilling stress increased the time to 50 % emergence
(E50) and mean emergence time (MET) (Fig. 1), electro-
lyte leakage (Fig. 7b), superoxide dismutase (SOD), cata-
lase (CAT) and ascorbate peroxidase (APX) activities
(Fig. 9), and decreased the emergence energy (EE), final
emergence (FEP) (Fig. 2), coefficient of uniformity of
emergence (CUE) (Fig. 3), shoot and root length (Fig. 4),
seedling fresh and dry weights (Fig. 5), leaf and root score
(Fig. 6), RWC (Fig. 7a) and starch metabolism (Fig. 8)
than at optimal temperature.
Seed-priming treatments with KCl reduced the E50
and MET, and increased the EE and CUE at both the
temperatures (Figs 1–3). Under both normal and stress
conditions, seed priming with 100 mg l)1 KCl was the
best treatment to reduce the E50 and MET and increase
the EE and CUE compared with other treatments
(Figs 1–3). Although seed-priming treatments improved
the FEP at low temperature, seed treatment with
100 mg l)1 KCl showed the best results; however, none
of the treatment could improve FEP at optimal tempera-
ture (Fig. 2b).
Seed priming with KCl also improved the shoot and
root length (Fig. 4), and seedling fresh and dry weights
at both the temperatures. There was no improvement in
leaf score at both the temperatures (Fig. 6a); however,
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Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 438–448

(a)
5 Chilling
E50 (days)
3
0
Control
(b)
8 Chilling
6
MET (days)
2
Fig. 1 Influence of pre-sowing KCl seed
treatments on the (a) time to 50%
0
emergence and (b) mean emergence time in
Control
maize at optimal and low temperatures
(±S.E.).
root score was improved only at optimal temperature
(Fig. 6). Maximum shoot and root length (Fig. 4), and
seedling fresh and dry weights (Fig. 5) under both opti-
mal and stress conditions, and root score (Fig. 6b) at
optimal temperature were recorded from seed priming
with 100 mg l)1 KCl.
Similarly, seed-priming treatments increased the RWC
and decreased electrolyte leakage at both low and optimal
temperatures (Fig. 7). Maximum RWC and minimum
electrolyte leakage under both stress and normal conditions
were recorded from 100 mg l)1 KCl treatment (Fig. 7b).
All KCl treatments improved the a-amylase activity and
soluble sugars at both the temperatures compared with
control (Fig. 8a). However, maximum sugars, and a-amy-
lases were recorded from 100 mg l)1 KCl treatment under
both conditions (Fig. 8).
Under both normal and stress conditions, seed priming
with KCl improved the superoxide dismutase (SOD)
(Fig. 9a), catalase (CAT) (Fig. 9b) and ascorbate per-
oxidase (APX) (Fig. 9c) activities compared with control.
However, maximum SOD, CAT and APX activities were
recorded from 100 mg l)1 KCl treatment under both con-
ditions (Fig. 9). Moreover, SOD, CAT and APX activities
were higher under chilling stress than at optimal tempera-
ture except at 50 mg L)1 where SOD was higher at optimal
temperature (Fig. 9).
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Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 438–448
Chilling Tolerance in Maize
Optimum temperature
50 100 150
Optimum temperature
50 100 150
Pre-sowing KCl (mg l–1) seed treatments
Parallels drawn between various determinations of
seedling growth, general metabolites and antioxidant
activity revealed that amylase activity was positively related
with root length and seedling fresh weight under optimum
conditions and with seedling dry weight under both opti-
mal and stress conditions (Table 1). Soluble sugars were
positively correlated with shoot length under optimal
conditions, and with root length and seedling dry weight
under both optimal and stress conditions. RWC was only
correlated with shoot length under optimal conditions,
and with seedling fresh and dry weight under stress con-
ditions (Table 1). SOD was positively correlated with root
and shoot lengths and seedling fresh weight under both
optimal and stress conditions. CAT was correlated with
root and shoot lengths and seedling fresh weight at opti-
mal temperature and with seedling dry weight under both
optimal and stress conditions (Table 1). Likewise, APX
was correlated with seedling dry weight under both
optimal and stress conditions (Table 1).
Discussion
Seed priming with KCl was effective enough to improve
the chilling tolerance in hybrid maize principally by the
activation of the antioxidant system (Fig. 9), although
starch metabolism was also improved by KCL under both
441
Farooq et al.

(a) 65 Optimum temperature

Chilling
52

39
EE (%)

26

13

0
Control 50 100 150

(b) Optimum temperature

Chilling
100

80
FEP (%)

60

40

20

Fig. 2 Influence of pre-sowing KCl seed

0 treatments on the (a) energy of emergence
Control 50 100 150
and (b) final emergence percentage in maize
Pre-sowing KCl (mg l–1) seed treatments at optimal and low temperatures (±S.E.).

1 Optimum temperature
Chilling

0.75
CUE

0.5

0.25

Fig. 3 Influence of pre-sowing KCl seed

0
treatments on the coefficient of uniformity of
Control 50 100 150
emergence (CUE) in maize at optimal and low
Pre-sowing KCl (mg l–1) seed treatments temperatures (±S.E.).

normal and stress conditions (Fig. 8). Among all, treat-
ment with 100 mg l)1 KCl was the most suitable to sig-
nificantly improve the emergence (Figs. 1–3), early
seedling growth (Figs 4 and 5), relative water content and
membrane stability (Fig. 7); starch metabolism (Fig. 8)
and antioxidant activities (Fig. 9).
Potash acts as a co-factor in many enzymes involved
in starch metabolism (Taiz and Zeiger 2006). Hence,
seed priming with KCl improved the starch metabolism.
Increase in starch metabolism and soluble sugars
(glucose, fructose and sucrose) has been found to be a
good indicator of freezing tolerance (Sasaki et al. 1998).
Membrane stability was significantly reduced as indi-
cated by the increased level of electrolyte leakage under
chilling stress (Fig. 7b). However, seed priming
improved membrane thermostability by reducing the

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442 Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 438–448
 Chilling Tolerance in Maize

(a) 25 Optimum temperature

Chilling
20

Shoot length (cm)

15

10

0
Control 50 100 150

(b) 20 Optimum temperature

Chilling
16
Root length (cm)

12

Fig. 4 Influence of pre-sowing KCl seed

0
treatments on the (a) shoot and (b) root
Control 50 100 150
length in maize at optimal and low tempera-
tures (±S.E.). Pre-sowing KCl (mg l–1) seed treatments

(a) 200 Optimum temperature

Chilling
Seedling fresh weight (mg)

150

100

50

0
Control 50 100 150

(b) Optimum temperature

32 Chilling
Seedling dry weight (mg)

24

16

Fig. 5 Influence of pre-sowing KCl seed

0
treatments on the (a) seedling fresh weight
Control 50 100 150
and (b) seedling dry weight in maize at
optimal and low temperatures (±S.E.). Pre-sowing KCl (mg l–1) seed treatments

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Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 438–448 443
Farooq et al.

(a) Optimum temperature

3
Chilling
2.5

2
Leaf score

1.5

0.5

0
Control 50 100 150

(b) Optimum temperature

8
Chilling

6
Root score

Fig. 6 Influence of pre-sowing KCl seed

0
treatments on the (a) leaf score and (b) root
Control 50 100 150
score in maize at optimal and low tempera-
Pre-sowing KCl (mg l–1) seed treatments tures (±S.E.).

(a) Optimum temperature

80 Chilling

60
RWC (%)

40

20

0
Control 50 100 150

(b) Optimum temperature

25 Chilling
Electrolyte leakage (%)

20

15

10

5
Fig. 7 Influence of pre-sowing KCl seed
0 treatments on the (a) relative water content
Control 50 100 150
(RWC) and (b) Electrolyte leakage in maize at
Pre-sowing KCl (mg l–1) seed treatments optimal and low temperatures (±S.E.).

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444 Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 438–448
 Chilling Tolerance in Maize

(a) 12 Optimum temperature

Chilling

α-amylase activity (unit)*

10

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

24 Optimum temperature
Chilling
18

12

Fig. 8 Influence of pre-sowing KCl seed

treatments on the (a) a-amylase activity and 6
(b) Soluble sugars in maize at optimal and
low temperatures (±S.E.). *One unit of the
0
enzyme’s activity is the amount of enzyme
Control 50 100 150
which released 1 lmol of maltose by 1ml
original enzyme solution in 1 min. Pre-sowing KCl (mg l–1) seed treatments

electrolyte leakage significantly (Fig. 7b). Enhanced elec- plants under chilling stress (Hérouart et al. 1991). ROS
trolyte leakage was considered to be a symptom of in plants are scavenged by a variety of antioxidant
stress-induced membrane damage and deterioration enzymes and/or lipid-soluble and water-soluble mole-
(Feng et al. 2003). However, K treatments decreased the cules (Foyer et al. 1994). Of these, antioxidant enzymes
electrolyte leakage by improving the membrane stability are the most effective against oxidative damage
as was reported previously (Premachandra et al. 1991). (Halliwell and Gutteridge 1999). The role of K in low-
Under low temperature stress, RWC were also decreased ering ROS production by reducing NAD(P)H oxidase
significantly (Fig. 7a), possibly because of decreased activity and maintaining photosynthetic electron trans-
metabolites and osmotica available to hold the water port (Cakmak 2005) justifies the activation of the
within the cells. The role of K in osmotic adjustment antioxidation system by KCl treatments. The present
and binding the tissue to water potential is very clear. study also suggests that enzymatic antioxidant activities
Increased K contents in chilling-tolerant plant species of maize seedlings were substantially induced during
can be correlated with osmoregulation-based mainte- chilling stress compared with those grown at normal
nance of tissue water status (Taiz and Zeiger 2006). temperature. This implied that the maize hybrid
Improved chilling tolerance was mainly attributed to Hycorn 8288 has the ability to activate its antioxidant
the enhanced capacity of the antioxidant system, main- system upon exposure to chilling and possibly under
tenance of high tissue water contents and carbohydrate other stresses.
metabolism as is supported by their positive relation In conclusion, seed priming with KCl can improve the
with seedling attributes (Table 1). low temperature tolerance in hybrid maize mainly by the
Exposure of plants to certain environmental stresses activation of antioxidant enzymes activities, indicating it
quite often leads to ROS production (Munne-Bosch to be a major metabolic event during tolerance to low
and Penuelas 2003), which may react with proteins, lip- temperature. Performance of hybrid maize was also
ids and DNA causing oxidative damage and impairing improved at normal temperature seed priming, with
the normal cellular functions (Foyer and Fletcher 2001). KCl 100 mg l)1 being the most effective under both
Increased expression of antioxidants occurs in several conditions.

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Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 438–448 445
Farooq et al.

(a)
Optimum temperature
SOD (unit g–1 protein) *
36
Chilling
30
24
18
12
6
0
Control 50 100 150
(b)
Optimum temperature
CAT (µmol min–1 g–1 protein)

20
Chilling
16

12

0
Control 50 100 150
(c)
Optimum temperature
APX (µmol min–1 g–1 protein)

15
Chilling
12

9
Fig. 9 Influence of pre-sowing KCl seed
6 treatments on the (a) superoxide dismutase
(SOD), (b) catalase (CAT) and (c) ascorbate
3 peroxidase (APX) in maize at optimal and low
temperatures (±S.E.). *One unit of SOD
0
activity is equivalent to the volume of extract
Control 50 100 150
needed to cause 50 % inhibition of the
Pre-sowing KCl (mg l–1) seed treatments colour reaction.

Table 1 Relationships of general metabolites and antioxidant activity with some growth attributes of seedlings as affected by seed priming with
KCl in hybrid maize under optimal and stress conditions (n = 4)

Correlation coefficient (r)

Shoot length Root length Seedling FW Seedling DW

Optimum Chilling Optimum Chilling Optimum Chilling Optimum Chilling

Variables temp. stress temp. stress temp. stress temp. stress

General metabolism
a-amylase activity 0.839 ns 0.453 ns 0.893* 0.705 ns 0.880* 0.710 ns 0.999*** 0.971**
Soluble sugars 0.896* 0.722 ns 0.927* 0.879* 0.922 ns 0.828 ns 0.988** 0.990***
RWC 0.882* 0.622 ns 0.860 ns 0.847 ns 0.872 ns 0.907* 0.842 ns 0.923*

Antioxidant activities
Superoxide dismutase 0.915* 0.944* 0.956* 0.972** 0.944* 0.976** 0.876 ns 0.718 ns
Catalase 0.899* 0.637 ns 0.939* 0.842 ns 0.930* 0.830 ns 0.993*** 0.998***
Ascorbate peroxidase 0.770 ns 0.580 ns 0.801 ns 0.777 ns 0.797 ns 0.732 ns 0.954* 0.989**

Significant at ***P < 0.001, **P < 0.01, *P < 0.05 and ns, non-significant.
RWC, relative water contents; FW, fresh weight; DW, dry weight.

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446 Journal compilation ª 2008 Blackwell Verlag, 194 (2008) 438–448

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