J Agro Crop Sci (2016) ISSN 0931-2250

SALINITY STRESS

Redox halopriming: A Promising Strategy for Inducing Salt

Tolerance in Bread Wheat
H.F. Maswada1 & N.I.K. Abd El-Kader2
1 Agricultural Botany Department, Faculty of Agriculture, Tanta University, Tanta, Egypt
2 Soil and Water Department, Faculty of Agriculture, Tanta University, Tanta, Egypt

Keywords Abstract
halopriming; nitric oxide; salinity tolerance;
wheat New strategies to enhance growth and productivity of food crops in saline soils
represent important research priorities. This study has investigated the role of
Correspondence certain priming techniques to induce salt tolerance of bread wheat. Wheat grains
Dr H. F. Maswada were soaked in 0.2 mM sodium nitroprusside as nitric oxide donor (redox prim-
Tanta University
ing), diluted sea water (halopriming) and the combination of both (redox halo-
Tanta, 31527
priming). Grains were also soaked in distilled water (hydropriming); in addition,
Egypt
Tel.:+20 10 11 48 65 80 untreated grains were taken as control. Our results indicated that priming treat-
Fax: +20 40 34 55 570 ments significantly improved all growth traits and increased leaf pigments con-
Email: hanafey2000@gmail.com centration as compared to the control. Priming treatments markedly enhanced
membrane stability index, proline, total soluble sugars and K+ concentration with
Accepted December 22, 2014 simultaneous decrease in the concentration of Na+ and malondialdehyde
(MDA). Furthermore, yield and yield-related traits such as plant height, spike
doi:10.1111/jac.12123
length, total number of tillers, 1000-grain weight, straw and grain yield consider-
ably affected by priming treatments. Moreover, the grain yield of both genotypes
was positively affected by redox halopriming treatment. However, the extent of
enhancement was more prominent in Gemmiza-9 (salt sensitive) than that in
Sakha-93 (salt-tolerant). Overall, this study clearly indicated that redox halopri-
ming treatment is a promising and handy technique to induce salinity tolerance
of wheat genotypes.

Introduction
By the end of year 2050, it is expected that world popula-
tion will increase to reach about 10 billion. On the other
hand, food productivity is decreasing due to the effect of
various abiotic and biotic stresses which cause losses worth
hundreds of million dollars each year. Thus, it is important
to minimize these losses to cope with the increasing food
requirements (Mahajan and Tuteja 2005).
Salinity is one of the major constraints to crop produc-
tivity due to the negative effects on plant growth, ion bal-
ance and water relations (Munns and Tester 2008). The
negative impact of soil salinity on the crop yields is related
to the abundance of Na+, where excess of Na+ is toxic to
plants (Zhang et al. 2010). Results that have been reported
previously by Kingsbury and Epstein (1986) indicated that
solutions with high concentrations of Na+ were more toxic
to bread wheat than those contain high concentrations of
Cl
. Moreover, the rate of Na+ uptake from the soil is con-
sidered to be the major cause of salinity toxicity and ion-
specific damage (Maathuis and Sanders 2001, Tester and
Davenport 2003). The availability of many essential nutri-
ents for plants was decreased due to high concentration of
Na+ in the soil. In addition, Na+ may directly compete with
many essential ions such as K+, Mg2+ and NHþ 4 on uptake
sites (Shabala and Munns 2012). Therefore, it has been
reported that reducing Na+ accumulation in crop plants is
considered to be one of the important mechanisms to
improve their tolerance to salt stress (Kingsbury and
Epstein 1986, Zhang et al. 2010).
There are several practical ways to minimize the con-
straints of salt stress on crop plants, such as seed priming
(Shabala and Munns 2012). Seed priming is one of the
effective seed invigoration methods to ameliorate the envi-
ronmental stresses, such as water deficit, high salinity,
extreme temperature, submergence, etc. (Jisha et al. 2013).

© 2015 Blackwell Verlag GmbH, 202 (2016) 37–50 37

Maswada and Abd El-Kader
It is a pre-sowing seed management technique that partially
hydrates seeds without radicle emergence (Chen and Arora
2013). In other words, seed priming, a controlled hydration
process, involves uptake of water by seed followed by dry-
ing to initiate pre-germinative physiological and biochemi-
cal activities (Manonmani et al. 2014). The risk of poor
stand establishment of plants under drought and salt stress
can be reduced by seed priming that permit more uniform
growth under saline soil conditions (Kaya et al. 2006).
Rapid seed germination and stand establishment are critical
factors for crop production under salt-affected soil condi-
tions (Ashraf and Foolad 2005). Therefore, the adverse
effects of salt stress can be alleviated by seed priming (Chen
and Arora 2013). However, there are different types of seed
priming such as hydropriming, halopriming, thermopri-
ming, biopriming, redox, nutrients and hormonal priming
(Ashraf and Foolad 2005, Imran et al. 2013, Jisha et al.
2013).
Over the past decade, the effect of nitric oxide (NO), a
reactive nitrogen species (RNS), on the biological systems
has been investigated for its biological importance, which
has been termed the ‘molecule of the year’ in 1992 (Kosh-
land 1992). The dual role of NO role of NO has been estab-
lished either as a protective or a toxic factor depending on
its concentration in the plant cell (Lamattina et al. 2003).
NO may act as pro-oxidant, as well as an antioxidant in
plants (Corpas et al. 2011). In addition, it was reported
that NO has a physiological role in regulation of plant
growth and development (Kopyra and Gw
o
zd
z 2004).
Recently, NO plays a crucial role in plant resistance to vari-
ous abiotic stresses such as salinity, drought, heavy metals,
heat and chilling (Siddiqui et al. 2010). For example, seed
priming or foliar spray with SNP, as NO donor, enhanced
tolerance of rice plants to drought stress (Farooq et al.
2009). Recent studies revealed that exogenous NO, as foliar
spray or pre-sowing (redox priming), plays systemic signal-
ling roles (Molassiotis et al. 2010, Jisha et al. 2013) in alle-
viating the oxidative damage caused by salinity in various
crops such as barley (Li et al. 2008), chickpea (Sheokand
et al. 2010), cucumber (Fan et al. 2013), maize (Zhang
et al. 2006), rice (Habib et al. 2010), soya bean (Simaei
et al. 2012), tomato (Wu et al. 2011) and wheat (Ruan
et al. 2002, Zheng et al. 2009, Kausar and Shahbaz 2013).
Halopriming refers to pre-sowing seed soaking in solu-
tions of high ionic strength. Seed halopriming was fre-
quently conducted with inorganic salts such as NaCl, CaCl2
and CaSO4, KNO3 with concentrations ranging from 50 to
200 mM (Shabala and Munns 2012). Several studies were
performed to investigate the effect of seed halopriming on
imparting salt tolerance of various crop plants such as
canola (Farhoudi and Sharifzadeh 2006), maize (Ashraf
and Rauf 2001), melon (Sivritepe et al. 2003), mung bean
(Saha et al. 2010), tomato (Nawaz et al. 2011) and wheat
38
(Afzal et al. 2008, Jamal et al. 2011, Abbasdokht and
Edalatpisheh 2013).
Standard sea water contains, Cl
, Na+, SO
2 2+
4 , Mg ,
2+ +

Ca , K and HCO3 (Aldesuquy and Ibrahim 2001, Mille-
ro et al. 2008), with high concentrations of sodium and
chloride ions in particular. Up to date, no studies have
been conducted to illustrate the influence of using sea
water in seed priming to promote salt tolerance of crop
species. Thus, it is worthy to investigate the potential role
of priming technique with diluted sea water (halopriming);
nitric oxide (redox priming) and both together (redox
halopriming) to induce salt tolerance of wheat. Halopri-
ming, redox priming and redox halopriming are consid-
ered to be types of defence priming. Defence priming
describes a process that immunity-like responses to a stress
are accelerated or enhanced by a prior stimulation (Molas-
siotis et al. 2010).
To investigate the ability of halopriming or redox prim-
ing treatments, individually or in combination, as a prom-
ising technique to induce salinity tolerance of bread wheat,
the main objective of this study is to improve grain yield of
bread wheat under salt-affected soil via simple and handy
technique that could be used by farmers in Egypt and other
similar countries suffer from salt-affected soils.
Materials and Methods
Plant material and experimental site
Two wheat (Triticum aestivum L.) genotypes, Sakha-93 as
salt-tolerant and Gemmiza-9 as salt sensitive, were selected
based on the previous studies of El-Hendawy et al. (2005)
and El-Lethy et al. (2013). The tested genotypes were
obtained from the National research centre at Dokki, Giza,
Egypt. The experiment was conducted at Agricultural Fac-
ulty Farm, Tanta, Egypt during 2012–2013. Data on the
weather during the whole course of study are given in
Table 1. In addition, EC, pH and the concentration of
cations (Na+, K+, Ca2+ and Mg2+) and anions (Cl
, SO
2 4 ,
CO
2

3 and HCO3 ) in the experimental soil are shown in
Table 2.
Treatments
Priming treatments included halopriming (diluted sea
water, EC = 9.00 dS m
1), redox priming [0.2 mM sodium
nitroprusside (SNP) as nitric oxide (NO) donor] and redox
halopriming (diluted sea water +0.2 mM SNP,
EC = 9.25 dS m
1). In addition, hydropriming (distilled
water) and untreated dry grains were taken as a control.
Sea water (EC = 62.03 dS m
1) was taken from the Medi-
terranean Sea in Baltim town (Kafr El-Sheikh Governorate,
Egypt). Sea water was diluted with distilled water to the
© 2015 Blackwell Verlag GmbH, 202 (2016) 37–50
 Inducing Salt Tolerance in Bread Wheat

Table 1 Meteorological data for El-Gharbyia area during the course of study

Monthly temperature (°C)

Total monthly Mean relative
Month Maximum Minimum Mean rainfall (mm) humidity (%)

November 2012 24.68 14.2 19.44 2.0 57.72

December 2012 21.75 11.27 16.51 18.0 53.67
January 2013 19.68 7.01 13.35 5.0 55.76
February 2013 22.42 8.39 15.41 2.9 46.16
March 2013 27.75 10.96 19.36 1.1 37.30
April 2013 27.72 11.87 19.80 0.0 39.86

Source: Meteorology station at Kotour, El-Gharbyia Governorate, Egypt, lat. 30°470 N and long 31°070 , elevation 15 cm.

Table 2 EC, pH and concentration of cations and anions of experimental soil and diluted sea water

Cations (meq l
1) Anions (meq l
1)

EC (dS m
1) pH Na+ K+ Ca2+ Mg2+ Cl
SO2

4 CO2

3 HCO

3

Diluted Sea water 9.00 6.62 39.46 0.81 27.56 22.17 46.52 38.42 0.00 5.06
Experimental Soil* 2.78 8.09 9.31 0.23 11.14 7.12 14.17 10.19 0.00 3.44

Soil analysis for pH based on soil: water extract (1 : 2.5), while EC and ions based on soil: water extract (1 : 5).
Soil texture was clay loam.

required concentration (EC = 9.00 dS m
1). The chemical
analysis of diluted sea water used in this study is given in
Table 2. For each priming treatment, a weighed quantity
(200 g) of wheat grains was soaked in 1.0 l of distilled
water/respective solution for 10 h. After soaking, grains
were removed, rinsed thoroughly with distilled water and
redried near to their original weight with forced air at
27  2 °C under shade (Farooq et al. 2013). The treated
and untreated grains were sealed in polythene bags and
stored in a refrigerator until use (Jafar et al. 2012).
Crop husbandry and experimental design
Prior to seedbed preparation, the experimental soil was
ploughed twice and randomized soil samples were taken
for analysis. After a week, the soil was levelled and divided
into three sections (blocks). Each block was divided into
more than 10 plots with a net plot area of 2.0 9 2.0 m.
Nitrogen, phosphorus and potassium fertilizers were
applied at the rate of 180, 75 and 120 kg per hectare in the
form of urea (46 % N), mono super-phosphate (15.5 %
P2O5) and potassium sulphate (48 % K2O), respectively.
All phosphorus and potassium and one-third of nitrogen
fertilizers were applied as basal dose during land prepara-
tion, while remaining nitrogen was applied with the first
and second irrigation in equal splits. Wheat genotypes were
sown manually on November 2012 using seed rate of
125 kg ha
1 (50 g per plot). In addition to soaking irriga-
tion, five surface irrigations were given during the whole
growing season at tillering, jointing (stem elongation),
booting, flowering and milking stages, respectively. The
field was kept weed-free by hoeing and manual weeding.
The wheat crop was harvested manually on April 2013. The
experiment was designed in completely randomized block
design (CRBD) in a factorial arrangement with three repli-
cations.
Growth analysis
Ten plant samples from each plot were collected at 30, 60
and 90 days after sowing (DAS), in other words at tillering,
stem elongation and flowering stages, respectively, then
divided into roots, leaf blades and aerial non-leaf blades to
estimate growth parameters. The dry mass of roots and
shoots (stems and leaves in addition to spikes at 90 DAS)
was determined at 70 °C for 48 h, while leaf area (LA) was
estimated using the equation LA = 0.75 9 leaf length 9
leaf width as described by Chanda and Singh (2002). In
addition, relative growth rate (RGR), unit leaf rate (ULR),
leaf area ratio (LAR) and specific leaf area (SLA) parame-
ters were calculated according to Hunt et al. (2002).
Physiological and biochemical attributes
At flowering stage, 10 plant samples were collected from
each plot, and flag leaves of the main stem of each plant
were separated to estimate physiological and biochemical
parameters. Leaf pigments, membrane stability index
(MSI), lipid peroxidation, proline and total soluble sugars
(TSS) were determined spectrophotometrically using
UV-VIS Spectrophotometer (Model SM1200; Randolph,
NJ, USA).

© 2015 Blackwell Verlag GmbH, 202 (2016) 37–50 39

Maswada and Abd El-Kader
Leaf pigments
Fresh flag leaves (0.1 g) were cut into small pieces (approx-
imately 1 9 1 mm) and put in brown glasses containing
20 ml of cold 80 % acetone for 24 h in a refrigerator
(4 °C) and then filtered. The absorbance of the filtrate was
read at 470 nm for the carotenoids, and at 646 and 663 nm
for chlorophyll a and b, respectively. Chlorophyll and
carotenoid concentrations were calculated using extinction
coefficient and equations described by Wellburn (1994).
The same protocol was carried out using cold methanol/
HCl/water (90 : 1 : 1, v : v : v), instead of cold 80 % ace-
tone, and the absorbance of filtrate was read at 529 and
650 nm for anthocyanin determination (Sims and Gamon
2002). Anthocyanin was calculated using the corrected
absorbance (AA = A529–0.288A650) and a molar absor-
bance coefficient for anthocyanin at 529 nm of
30 000 l mol
1 cm
1 (Murray and Hackett 1991).
Membrane stability index and lipid peroxidation
Membrane stability index was determined with previously
described protocols (Shanahan et al. 1990, Yang et al.
1996). Five segments (0.5 9 0.5 cm) of the flag leaves for
each replicate were cut and put in test tubes each having
15 ml distilled water, vortexed for 3 s and kept at 4 °C for
24 h. The initial conductivity (EC1) was measured after
bringing sample to 25 °C using CMD 830 WPA conductiv-
ity metre. The samples were then autoclaved for 20 min,
cooled to 25 °C, and final conductivity (EC2) was recorded.
MSI was calculated as follows: MSI (%) = [1
(EC1/
EC2)] 9 100.
Lipid peroxidation was determined by measuring the
amount of malondialdehyde (MDA) according to the
method of Davenport et al. (2003). Fresh flag leaves (0.2 g)
were homogenized with 2 ml of 5 % (w/v) trichloroacetic
acid in an ice bath and centrifuged at 10 000 g for 10 min
at 4 °C. Two ml supernatant mixed with 2 ml of 0.67 %
(w/v) thiobarbituric acid was incubated in boiling water for
30 min, then cooled and centrifuged. The absorbance of
reaction supernatant was assayed at 450, 532 and 600 nm.
The MDA concentration was calculated based on the fol-
lowing formula: MDA (nmol g
1 Fw) = [6.45 9 (A532

A600)
(0.56 9 A450)] 9 V/W, where V = volume
(ml); W = weight (g).
Proline and total soluble sugars
Proline concentration of fresh flag leaves was estimated
using 3 % (w/v) aqueous sulphosalicylic acid and ninhy-
drin reagent according to Bates et al. (1973). Proline con-
centration (lg g
1 Fw) was quantified using L-proline as
standard.
40
Total soluble sugars were estimated using anthrone
reagent (Watanabe et al. 2000) with slight modification. In
brief, flag leaf tissue (0.2 g) was extracted with 10 ml etha-
nol (95 % v/v) for 24 h and then filtered. The filtrate
(1 ml) was mixed with freshly prepared anthrone reagent
(3 ml) and the tubes heated at 100 °C for 10 min. The
reaction was terminated by quick cooling on ice. The
absorbance was measured at 620 nm. The TSS
(mg g
1 Fw) were quantified using glucose as standard.
Sodium and potassium ions
At flowering stage, 10 plant samples from each replicate for
each treatment were collected and divided into roots and
shoots (stems, leaves and spikes) and then dried in oven at
70 °C for 48 h. Fine ground roots and shoots (0.2 g) were
digested with concentrated H2SO4 (5 ml) and 80 % per-
chloric acid (1 ml). Digested material diluted by distilled
water to 100 ml. Na+ and K+ concentrations estimated using
PFP7 Flame photometer (Temminghoff and Houba 2004).
Agronomic and yield attributes
Wheat plants were harvested after physiological maturity
(at Zadoks scale 92, Zadoks et al. 1974) according to
El-Hendawy et al. (2009). One m2 of each plot was manu-
ally harvested, and 10 plant samples were randomly chosen
from each plot to estimate plant height, number of total til-
lers per plant, spike length, number of spikelets and grains
per spike. In addition, 1000-grain weight, grain and straw
yields and harvest index were also estimated.
Statistical analysis
All measurements were analysed using computer software
MSTAT-C (a computer based Statistical software packages
developed by the Crop and Soil Sciences Department,
Michigan State University, East Lansing, MI, USA). Mean
values and significance were determined by Duncan’s multi-
ple range tests, at P ≤ 0.05 (Gomez and Gomez 1984). The
mean values with different letters are considered signifi-
cantly different.
Results
Growth attributes
In both wheat genotypes, root dry mass (RDM) and shoot
dry mass (SDM) at 30 DAS (tillering stage) were not signif-
icantly affected by priming treatments. While, at 60 DAS
(stem elongation stage) and 90 DAS (flowering stage),
RDM and SDM were significantly increased by applying
seed priming (Table 3). However, this increase was more
© 2015 Blackwell Verlag GmbH, 202 (2016) 37–50
 Inducing Salt Tolerance in Bread Wheat

Table 3 Effect of priming treatments on root dry mass, shoot dry mass and leaf area of wheat genotypes at tillering (30 DAS), stem elongation (60
DAS) and flowering (90 DAS) stages under saline conditions

Root dry mass (g plant
1) Shoot dry mass (g plant
1) Leaf area (cm2)

Genotype (G) Priming (P) 30 DAS 60 DAS 90 DAS 30 DAS 60 DAS 90 DAS 30 DAS 60 DAS 90 DAS

Sakha-93 P0 0.015 a 0.184 c 0.291 c 0.135 b 1.119 b 5.420 cde 8.20 de 20.88 bc 21.52 b
P1 0.012 a 0.240 bc 0.344 c 0.139 b 1.235 ab 6.686 bc 8.53 cd 20.94 bc 21.95 b
P2 0.016 a 0.287 abc 0.439 abc 0.147 b 1.384 ab 6.949 bc 8.86 cd 24.72 a 24.73 ab
P3 0.014 a 0.381 a 0.537 ab 0.146 b 1.452 ab 9.169 a 10.16 a 22.91 abc 23.94 b
P4 0.015 a 0.297 ab 0.604 a 0.154 b 1.624 a 8.356 ab 9.90 ab 23.36 abc 24.08 b
Gemmiza-9 P0 0.013 a 0.239 bc 0.277 c 0.142 b 1.219 ab 4.339 e 7.56 e 20.13 c 22.65 b
P1 0.012 a 0.274 bc 0.286 c 0.153 b 1.479 ab 4.770 de 10.02 ab 23.50 abc 23.92 b
P2 0.016 a 0.307 ab 0.337 bc 0.178 a 1.328 ab 6.694 bc 10.62 a 22.30 abc 24.23 b
P3 0.014 a 0.340 ab 0.364 bc 0.145 b 1.352 ab 6.340 cd 9.24 bc 23.97 ab 24.25 b
P4 0.013 a 0.301 ab 0.365 bc 0.146 b 1.562 a 6.816 bc 10.44 a 25.77 a 27.56 a
F values and level G 0.468 ns 0.508 ns 10.45 ** 3.136 ns 0.111 ns 17.82 ** 7.014 * 0.731 ns 4.350 ns
of significance P 2.260 ns 5.820 ** 4.453 * 2.784 ns 3.181 * 9.303 ** 22.13 ** 4.291 * 4.406 *
G9P 0.324 ns 0.626 ns 1.217 ns 2.021 ns 0.721 ns 1.405 ns 10.36 ** 2.032 ns 1.258 ns

Values represent the means of three replicates of each treatment (10 plant samples of each plot). Means values within each column followed by the
same letter are not significantly different (P ≤ 0.05), according to Duncan’s multiple range test. ** and *: significant at 0.01 and 0.05 probability lev-
els, respectively. riming treatments include control ‘P0’, hydropriming ‘P1’, redox priming ‘P2’, halopriming ‘P3’ and redox halopriming ‘P4’.
ns, non-significant; DAS, days after sowing.

prominent in Sakha-93 (salt-tolerant genotype) than that
in Gemmiza-9 (salt-sensitive genotype). Maximum RDM
and SDM of Sakha-93 at 60 and 90 DAS were recorded by
redox halopriming and halopriming treatments. Priming
treatment significantly affected leaf area (LA) of both geno-
types at all tested phenological stages (Table 3). However,
the significant difference between both genotypes appeared
only at tillering stage, where LA was higher in Gemmiza-9
than that in Sakha-93. Maximum LA of Gemmiza-9 tiller-
ing stage was obtained by redox halopriming treatment.
In both genotypes, RGR, ULR, LAR and SLA from 30 to
90 DAS were significantly increased by seed priming, espe-
cially halopriming on Sakha93 and redox halopriming on
Gemmiza-9 (Table 4). Furthermore, RGR and ULR were
significantly higher in Sakha-93 than those in Gemmiza-9,
but there was not significantly difference between both
genotypes in case of LAR and SLA.
Physiological and biochemical parameters
The concentration of chlorophyll (Chl) a, b, carotenoids
and anthocyanin in flag leaves of both genotypes was sig-
nificantly increased by priming treatments, especially redox
halopriming, in both genotypes at flowering stage. While,
Chl a/b and Chl/carotenoids ratios were not significantly
affected (Table 5). Despite of the lack of the significant dif-
ferences in the concentration of leaf pigments between
untreated wheat genotypes, redox halopriming remarkably
increased the concentrations of Chl a, b and carotenoids in
salt-sensitive genotype (Gemmiza-9) than those in salt-tol-
erant genotype (Sakha-93). The same way was observed for
anthocyanin, but the enhancement was higher in salt-toler-
ant than that in salt sensitive (Table 5).
In both wheat genotypes, MSI % and the concentrations
of TSS and proline of flag leaves at flowering stage were sig-
nificantly higher in primed plants than non-primed plants.
By contrast, priming treatments markedly lowered MDA
concentration compared to the control (Table 6). How-
ever, maximum MSI was recorded by redox halopriming,
and redox priming treatments, while maximum proline
concentration was observed by redox priming treatment.
Moreover, maximum TSS concentration was achieved by
redox priming and redox halopriming for Sakha-93, and
Gemmiza-9, respectively. MSI %, TSS and proline concen-
trations were significantly higher in Sakha-93 than those in
Gemmiza-9, and oppositely in case of MDA concentration
(Table 6).
Application of priming treatments to wheat genotypes
resulted in a decrease of Na+ accumulation in the shoots
and roots as compared to the control plants, while K+ accu-
mulation and K+/Na+ ratio were significantly higher under
the same conditions (Table 7). In the shoots, the maximum
decline of Na+ concentration and increase of K+/Na+ ratio
were recorded by redox priming for Sakha-93 and redox
halopriming for Gemmiza-9. In addition, maximum K+
accumulation in the shoot of both genotypes was obtained
from redox halopriming, redox priming and halopriming
treatments. In the roots, the maximum decrease of Na+ as
well as the maximum increase of K+ and K+/Na+ ratio was
obtained by redox halopriming treatment. Na+
accumulation in the root of salt-tolerant genotype was
higher than that in the root of salt-sensitive one, while K+

© 2015 Blackwell Verlag GmbH, 202 (2016) 37–50 41

Maswada and Abd El-Kader

Table 4 Effect of priming treatments on relative growth rate (RGR), unit leaf rate (ULR), leaf area ratio (LAR) and specific leaf area (SLA) of wheat
genotypes from 30 to 90 DAS under saline conditions

Genotype (G) Priming (P) RGR (g g
1 month
1) ULR (g m2 day
1) LAR (cm2 g
1) SLA (cm2 g
1)

Sakha-93 Control 3.641 bcd 68.29 cd 29.31 bc 48.63 abc

Hydropriming 3.838 abc 82.18 abc 29.90 bc 48.64 abc
Redox priming 3.805 abc 79.78 abc 28.92 bc 47.19 bc
Halopriming 4.104 a 100.76 a 33.11 ab 56.02 a
Redox halopriming 3.963 ab 93.81 ab 30.68 bc 49.56 abc
Gemmiza-9 Control 3.392 d 55.27 d 26.93 c 42.27 c
Hydropriming 3.403 d 52.38 d 32.80 ab 50.34 abc
Redox priming 3.588 cd 70.41 bcd 29.01 bc 47.50 bc
Halopriming 3.735 bc 71.04 bcd 31.08 abc 49.98 abc
Redox halopriming 3.790 abc 68.06 cd 34.95 a 54.17 ab
F values and level G 20.63 ** 21.53 ** 0.485 ns 0.577 ns
of significance P 5.741 ** 3.577 * 4.928 ** 3.365 *
G9P 0.596 ns 0.868 ns 2.589 ns 2.045 ns

Values represent the means of three replicates of each treatment (10 plant samples of each plot). Means values within each column followed by the
same letter are not significantly different (P ≤ 0.05), according to Duncan’s multiple range test. ** and *: significant at 0.01 and 0.05 probability
levels, respectively.
ns, non-significant; DAS, days after sowing.

Table 5 Effect of priming treatments on leaf pigments of fresh flag leaves of wheat genotypes at flowering stage (90 DAS) under saline conditions

Carotenoids Anthocyanin
Genotype (G) Priming (P) Chl a (mg g
1 Fw) Chl b (mg g
1 Fw) Chl a/b ratio (mg g
1 Fw) Chl/carotenoids (lmol g
1 Fw)

Sakha-93 Control 1.703 d 0.670 de 2.495 d 0.396 c 5.995 ab 0.219 de

Hydropriming 1.882 cd 0.717 bcd 2.620 bc 0.434 bc 5.992 ab 0.220 de
Redox priming 2.042 bc 0.769 bc 2.654 abc 0.462 b 6.085 a 0.273 b
Halopriming 1.916 c 0.719 bcd 2.664 abc 0.445 bc 5.927 ab 0.247 c
Redox halopriming 2.055 bc 0.768 bc 2.678 abc 0.478 ab 5.909 ab 0.302 a
Gemmiza-9 Control 1.684 d 0.612 e 2.754 a 0.393 c 5.849 b 0.207 e
Hydropriming 1.942 c 0.710 cd 2.736 ab 0.457 b 5.798 b 0.238 cd
Redox priming 2.172 ab 0.786 bc 2.764 a 0.495 ab 5.978 ab 0.245 c
Halopriming 2.076 bc 0.801 ab 2.593 cd 0.472 ab 6.099 a 0.242 cd
Redox halopriming 2.339 a 0.873 a 2.679 abc 0.530 a 6.064 a 0.276 b
F values and level G 8.154 * 3.546 ns 12.69 ** 7.391 * 0.361 ns 5.065 *
of significance P 16.20 ** 17.57 ** 1.943 ns 14.26 ** 1.721 ns 29.90 **
G9P 1.391 ns 4.146 * 5.816 ** 0.869 ns 3.767 * 3.104 *

Values represent the means of three replicates of each treatment (five plant samples of each plot). Means values within each column followed by the
same letter are not significantly different (P ≤ 0.05), according to Duncan’s multiple range test. ** and *: significant at 0.01 and 0.05 probability
levels, respectively.
ns, non-significant; DAS, days after sowing.

accumulation followed an opposite trend. By contrast, K+
accumulation in the shoot of salt-tolerant genotype was
higher than that in the shoot of salt-sensitive one, while
Na+ accumulation followed an opposite trend (Table 7).
Yield and yield-related traits
In both genotypes, priming treatments have significant
effects on all agronomic and yield attributes except for har-
vest index (Table 8). Under control conditions, there was
not significantly difference between wheat genotypes for
plant height, number of spikelets and grains per spike, and
straw yield. However, priming treatments markedly
increased these parameters in salt-sensitive genotype than
those in salt-tolerant one. In addition, salt-tolerant geno-
type possessed higher values of 1000-grain weight and har-
vest index than those in salt-sensitive genotype. Maximum
plant height was observed by halopriming and redox halo-
priming treatment in Sakha-93, and Gemmiza-9, respec-
tively. Redox priming was the best treatment for improving
number of total tillers for both genotypes, spike length for
Gemmiza-9 and straw yield for Sakha-93. While redox
halopriming was the best treatment for enhancing number
of spikelets and grains per spike for genotypes, 1000-grain

42 © 2015 Blackwell Verlag GmbH, 202 (2016) 37–50

 Inducing Salt Tolerance in Bread Wheat

Table 6 Effect of priming treatments on membrane stability index (MSI %) and the concentrations of malondialdehyde (MDA), proline and total
soluble sugars (TSS) of fresh flag leaves of wheat genotypes at flowering stage (90 DAS) under saline conditions

Genotype (G) Priming (P) MSI (%) MDA (nmol g
1 Fw) Proline (lg g
1 Fw) TSS (mg g
1 Fw)

Sakha-93 Control 21.61 e 9.27 de 11.38 g 22.88 de

Hydropriming 31.61 c 8.54 ef 14.20 f 24.99 c
Redox priming 36.05 a 8.30 ef 31.44 a 32.17 a
Halopriming 34.34 ab 8.54 ef 20.18 e 25.66 c
Redox halopriming 36.79 a 7.23 f 26.56 c 29.32 b
Gemmiza-9 Control 20.70 e 17.93 a 10.39 g 22.51 e
Hydropriming 27.72 d 16.21 b 11.57 g 22.89 de
Redox priming 34.53 ab 11.27 c 28.39 b 26.37 c
Halopriming 33.17 bc 15.98 b 19.87 e 24.69 cd
Redox halopriming 36.36 a 10.17 cd 24.14 d 32.43 a
F values and level G 9.457 ** 360.5 ** 24.74 ** 10.20 **
of significance P 117.3 ** 33.61 ** 364.1 ** 67.24 **
G9P 1.370 ns 15.62 ** 1.908 ns 13.99 **

Values represent the means of three replicates of each treatment (five plant samples of each plot). Means values within each column followed by the
same letter are not significantly different (P ≤ 0.05), according to Duncan’s multiple range test. ** and *: significant at 0.01 and 0.05 probability lev-
els, respectively.
ns, non-significant; DAS, days after sowing.

Table 7 Effect of priming treatments the concentrations of Na+, K+ as well as K+/Na+ ratio in the root and shoot of wheat genotypes at 90 DAS
(flowering stage) under saline conditions

Shoot Na+ Shoot K+ Root Na+ Root K+ Root K+/Na+

Genotype (G) Priming (P) (mg g
1 Dw) (mg g
1 Dw) Shoot K+/Na+ ratio (mg g
1 Dw) (mg g
1 Dw) ratio

Sakha-93 Control 6.017 cd 6.617 bc 1.100 ef 5.117 a 1.517 e 0.297 e

Hydropriming 5.617 e 6.883 b 1.227 d 4.817 b 1.727 e 0.360 e
Redox priming 5.110 f 8.117 a 1.590 a 4.667 bcd 3.183 c 0.683 c
Halopriming 5.543 e 7.967 a 1.437 c 4.550 de 3.183 c 0.697 c
Redox halopriming 5.500 e 8.250 a 1.500 b 3.767 g 3.617 b 0.963 b
Gemmiza-9 Control 6.610 a 5.450 d 0.823 h 4.733 bc 1.647 e 0.347 e
Hydropriming 6.300 b 5.567 d 0.883 g 4.583 cde 2.483 d 0.540 d
Redox priming 6.160 c 6.533 c 1.060 f 4.433 e 3.333 c 0.753 c
Halopriming 6.153 c 6.483 c 1.053 f 4.250 f 3.300 c 0.777 c
Redox halopriming 5.933 d 6.750 bc 1.140 e 3.533 h 4.717 a 1.337 a
F values and level G 530.2 ** 443.2 ** 1104.4 ** 79.89 ** 59.13 ** 66.78 **
of significance P 65.49 ** 82.29 ** 166.6 ** 196.3 ** 245.4 ** 239.3 **
G9P 12.25 ** 1.242 ns 13.245 ** 0.916 ns 11.94 ** 10.60 **

Values represent the means of three replicates of each treatment (dry shoot and root of 10 plant samples of each plot). Means values within each col-
umn followed by the same letter are not significantly different (P ≤ 0.05), according to Duncan’s multiple range test. ** and *: significant at 0.01
and 0.05 probability levels, respectively.
ns, non-significant; DAS, days after sowing.

weight and straw yield for Gemmiza-9. Similarly, maxi-
mum grain yield of both genotypes was observed from
redox halopriming; where the enhancement of grain yields
over control was 96.09 % and 36.61 % in Gemmiza-9 and
Sahka-93, respectively.
Discussion
Salinity is one of the most critical abiotic factors of the
growth and productivity of crop plants. Munns and Tester
(2008) reported that more than 45 million hectares of
irrigated land have been damaged by salt worldwide and
nearly 1.5 million hectares are taken out of production each
year due to high salinity levels in the soil. Thus, it is worthy
to enhance the performance of crop plants, especially food
crops, under saline condition. This study provides a prom-
ising strategy to enhance salt tolerance of bread wheat,
which is considered the main food crop worldwide.
Seed priming with nitric oxide (redox priming), diluted
sea water (halopriming) or the combination of both (redox

© 2015 Blackwell Verlag GmbH, 202 (2016) 37–50 43

Maswada and Abd El-Kader

Table 8 Effect of priming treatments on agronomic and yield attributes of wheat genotypes under saline conditions

No. total Spike No. 1000- Straw Grain

Genotype Priming Plant tillers length spikelets No. grains grain yield yield Harvest
(G) (P) height (cm) per plant (cm) per spike per spike weight (t ha
1) (t ha
1) index (%)

Sakha-93 P0 86.97 cd 4.10 bcd 8.35 cd 15.63 d 34.70 e 50.40 a 8.85 d 4.747 de 34.87 cd
P1 85.20 d 4.50 a-d 9.06 ab 16.27 cd 35.67 e 47.89 b 9.81 d 4.887 d 33.25 cde
P2 89.90 bcd 4.90 ab 8.07 d 16.81 bc 36.49 de 44.13 de 11.90 c 6.303 bc 34.66 cd
P3 92.00 bc 4.00 cd 9.02 ab 16.73 bcd 36.25 de 47.22 bc 9.63 d 6.067 c 38.68 ab
P4 90.50 bcd 3.70 d 8.69 abc 17.97 a 40.75 bc 48.71 ab 9.92 d 6.58 abc 40.33 a
Gemmiza-9 P0 86.93 cd 4.50 a-d 7.91 de 16.00 cd 36.67 cde 44.02 de 8.36 d 3.840 e 31.48 e
P1 89.07 bcd 3.90 cd 7.43 e 16.28 cd 37.10 cde 43.69 de 9.95 d 5.650 cd 36.23 bc
P2 91.10 bcd 5.10 a 9.23 a 17.52 ab 44.15 b 43.14 e 14.74 b 7.160 ab 32.68 de
P3 94.30 b 4.70 abc 8.94 ab 18.18 a 40.00 cd 44.00 de 12.75 c 5.950 c 31.80 de
P4 102.00 a 4.60 abc 8.64 bc 18.57 a 47.98 a 45.57 cd 16.97 a 7.530 a 33.47 e
F values and G 10.07 ** 3.765 ns 3.881 ns 8.558 ** 29.28 ** 66.04 ** 23.49 ** 2.502 ns 40.47 **
level of P 9.030 ** 3.493 * 8.549 ** 16.44 ** 14.82 ** 8.705 ** 24.29 ** 26.22 ** 2.345 ns
significance G9P 2.947 * 2.478 ns 17.84 ** 1.234 ns 2.544 ns 3.917 * 7.155 ** 3.369 * 13.13 **

Values represent the means of three replicates of each treatment (10 plant samples of each plot were harvested to estimate plant height, No. total til-
lers per plant, spike length, No. spikelets per spike and No. grains per spike, while 1000-grain weight, straw and grain yields were estimated from
plant samples that harvested from 1 m2 of each plot). Means values within each column followed by the same letter are not significantly different
(P ≤ 0.05), according to Duncan’s multiple range test. ** and *: significant at 0.01 and 0.05 probability levels, respectively. Priming treatments
include control ‘P0’, hydropriming ‘P1’, redox priming ‘P2’, halopriming ‘P3’ and redox halopriming ‘P4’.
ns, non-significant.

halopriming) has a positive impact in improving growth
traits including, RDM, SDM, LA, RGR, ULR, LAR and SLA
of both wheat genotypes under salt-affected soil. However,
applying redox halopriming has the major impact on
increase of RDM, SDM and LA in both genotypes as com-
pared to applying redox and halopriming, separately
(Table 3). In addition, redox halopriming followed by
redox priming has positive effects on RGR, ULR, LAR and
SLA (Table 4). Accordingly, seed priming with diluted sea
water and/or nitric oxide have positive effects on improv-
ing growth traits of salinized wheat plants. In the line of
our results, seed priming with inorganic salts (halopri-
ming) had positive effect for promoting growth and dry
matter accumulation of several plants (Table 9) such as,
canola (Farhoudi and Sharifzadeh 2006), maize (Ashraf
and Rauf 2001), melon (Sivritepe et al. 2003), mung bean
(Saha et al. 2010), tomato (Nawaz et al. 2011) and wheat
(Afzal et al. 2008, Jamal et al. 2011, Abbasdokht and Eda-
latpisheh 2013). In addition, seed priming with 0.1 mM
SNP, as NO donor, improved growth as indicated by an
increase in the dry matter accumulation of maize (Zhang
et al. 2006) and wheat (Zheng et al. 2009). Moreover, Kau-
sar and Shahbaz (2013) mentioned that foliar application
of NO (0.05, 0.1 and 0.15 mM) on wheat plants was found
to be effective in enhancing shoot and root fresh weights
and leaf area per plant under saline and non-saline condi-
tions. Accordingly, it is suggested that NO actively partici-
pates in growth regulation of plants. However, the
improvement of plant growth as response to exogenous
NO might be due to the higher activity of antioxidant
enzymes (Wu et al. 2011). Moreover, Kausar and Shahbaz
(2013) indicated that growth attributes of wheat plants
have been increased by application of NO due to an
increase of photosynthetic rate and stomatal conductance
(Table 9). Our results indicated that salt-tolerant genotype
has higher RDM, SDM, RGR and ULR than salt-sensitive
one. Similar results were obtained on salt-tolerant wheat
genotypes, where the growth rate and shoot biomass were
higher as compared to salt-sensitive ones (Rahnama et al.
2011). The growth and net assimilation rates are reliable
indicators to differentiate wheat genotypes for salt toler-
ance (Kumar et al. 2012).
Chlorophyll concentration was used as a sensitive indica-
tor of the cellular metabolic state of plants under salt stress
(Chutipaijit et al. 2011). Moreover, grown plants in saline
soils exhibit leaf chlorosis due to chlorophyll decay (Jiang
et al. 1994), where salinity increases the activity of chloro-
phyllase (Rao and Rao 1981) that degrades chlorophyll and
causing instability of pigment complexes (Singh and Dubey
1995). In this study, redox halopriming treatment has a
positive impact on chlorophyll (Chl) a, b, carotenoids and
anthocyanin concentrations (Table 5). In line with these
findings (Table 9), the negative impacts of salinity on Chl
a, total Chl and carotenoid concentrations, as well as Chl a/
b ratio in Aegiceras corniculatum leaves (Chen et al. 2014),
and anthocyanin concentration in the leaves of soya bean
(Simaei et al. 2012) were restored significantly by seed
treatment with 100 lM SNP. Furthermore, exogenous NO

44 © 2015 Blackwell Verlag GmbH, 202 (2016) 37–50


could apparently counteract the decay of chlorophyll in
wheat leaves caused by salt and osmotic stress (Ruan et al.
2002, Alavi et al. 2014). Moreover, treating maize seedlings
with 100 lM SNP remarkably increased chlorophyll con-
centration under salt stress (Zhang et al. 2006). Ali and Is-
mail (2014) reported that spraying salinized tomato plants
with 10 lM SNP significantly increased b-carotene and
anthocyanin concentrations in fruits. In addition, pre-trea-
ted seeds of mung bean with NaCl (50 mM) overcame the
adverse effect of salt stress by increasing growth and photo-
synthetic pigments (Chl a, b and carotenoids) of seedlings
(Saha et al. 2010) (Table 9). In this study, our results shed
some light on the effect of seed priming on increasing the
photosynthetic pigments in salt-sensitive genotype that
may help to improve plant performance under saline con-
ditions (Table 5). Consequently, we can deduce that
increasing photosynthetic pigments, due to seed priming,
might be one of the mechanisms that enabled salt-sensitive
wheat genotype to tolerate salt stress.
When plants are exposed to salinity or osmotic stress,
ROS begins to accumulate and causes deterioration of
membrane lipids, leading to an increase of MDA concen-
tration and membrane permeability (Gunes et al. 2007).
Lipid peroxidation, as indicated by lower levels of electro-
lyte leakage and MDA concentration, has been used as indi-
cator of salt-induced oxidative damage to the membranes
(Hernandez and Almansa 2002). Our results indicated the
effectiveness of redox halopriming on decreasing MDA and
improving MSI (Table 6); accordingly, the membrane
integrity can be considerably maintained by applying redox
halopriming treatment. In line with our findings (Table 9),
several studies have been conducted to investigate the effect
of the exogenous NO on the salt tolerance in A. cornicula-
tum (Chen et al. 2014), barley (Li et al. 2008), chickpea
(Sheokand et al. 2010), cucumber (Fan et al. 2013), maize
(Zhang et al. 2006), soya bean (Simaei et al. 2012), tomato
(Wu et al. 2011) and wheat (Ruan et al. 2002, Zheng et al.
2009, Alavi et al. 2014) via reducing lipid peroxidation
and/or improving MSI. Similarly, halopriming with inor-
ganic salts (NaCl) had a potential role in mitigating salt
and osmotic stresses by increasing MSI or reducing MDA
concentration in mung bean (Saha et al. 2010), or lowering
electrical conductivity of seed leachate in tomato (Nawaz
et al. 2011).
The accumulation of organic osmolytes (compatible sol-
utes) as well as inorganic ions plays an important role in
maintaining osmotic adjustment that enhances the growth
of plants under salt stress (Chen and Jiang 2010, Patade
et al. 2011). Compatible solutes acting as low molecular
weight chaperones and play an important role in protecting
cellular structures through scavenging ROS (Hasegawa
et al. 2000). It is suggested that proline serves as a selective
element for the tolerance of most plant species to different
© 2015 Blackwell Verlag GmbH, 202 (2016) 37–50
Inducing Salt Tolerance in Bread Wheat
environmental stresses (Ashraf and Foolad 2007). More-
over, the accumulation of proline is an adaptive mechanism
that improves the plant performance under salt stress (Khe-
dr et al. 2003). Redox and redox halopriming were the most
relevant treatments that significantly increased the concen-
tration of free proline and TSS in both wheat genotypes
compared to control (Table 6). The same findings, as a
result of exogenous NO, were reported in wheat (Ruan
et al. 2002, Zheng et al. 2009) and in tomato (Wu et al.
2011). In addition, halopriming with inorganic salts
improved salt tolerance either by the accumulation of pro-
line in canola (Farhoudi and Sharifzadeh 2006), melon
(Sivritepe et al. 2003) and mung bean (Saha et al. 2010), or
by the accumulation of TSS as reported by Nawaz et al.
(2011) for tomato and Afzal et al. (2008) for wheat
(Table 9).
Increase of K+ uptake with decease of Na+ accumulation
is amongst the important indicator of salinity tolerance
(Marschner 1995). Also, K+/Na+ ratio is considered as a
good indicator to assess plant tolerance to salinity (Ashraf
et al. 2010). Zhang et al. (2011) reported that salt stress
markedly increased Na+ concentration and reduced K+
accumulation and K+/Na+ ratio in wheat plants especially
during the late filling stage. The higher K+ concentration
and K+/Na+ ratio, with lower Na+ accumulation in the
roots and shoots of both wheat genotypes (Table 7), shed
some light on the reliability of using redox halopriming to
enhance salt tolerance. In line with our results, redox prim-
ing with NO (Zheng et al. 2009), as well as halopriming
with inorganic salts (Afzal et al. 2008, Jamal et al. 2011,
Jafar et al. 2012, Abbasdokht and Edalatpisheh 2013) main-
tained the ion balance between K+ and Na+, where priming
decreased the Na+ uptake and increased K+ accumulation
and K+/Na+ ratio in wheat plants grown under saline con-
ditions (Table 9). Moreover, priming with NaCl induced
salt tolerance of canola (Farhoudi and Sharifzadeh 2006),
and melon (Sivritepe et al. 2003) via increasing K+ and
Ca2+ concentration, with decreasing Na+ accumulation.
Also, Simaei et al. (2012) reported that treating soya bean
seedlings with 100 lM of SNP improved K+ uptake and
inhibited Na+ uptake of roots and shoots. The lower accu-
mulation of Na+ in the shoots of the salt-tolerant genotype
that accumulates higher Na+ in the roots (Table 7) pro-
vided evidence for the reduction of Na+ translocation from
roots to shoots that is considered as a mechanism of salt
tolerance (Boursier and L€auchi 1990).
The yield reduction of crop plants is most countable
effect of salinity in agriculture, where yield of most gylco-
phytic crops declined greatly due to salt stress (Hasanuzz-
aman et al. 2013). Seed priming with diluted sea water
and/or nitric oxide has positive effects on the agronomic
and yield attributes of both wheat genotypes (Table 8).
Similar results were reported by Jafar et al. (2012)
45
Maswada and Abd El-Kader

Table 9 Effects of halopriming with inorganic salts as well as exogenous nitric oxide to induce salt tolerance of wheat or other crop plants

Plant Treatment Effects Reference

Wheat Priming with KNO3 (-1.2 MPa) Improved dry matter accumulation. Increased Abbasdokht and
K+ concentration and K+/Na+ ratio; and Edalatpisheh (2013)
decreased Na+ accumulation.
Priming with CaCl2, Improved root and shoot dry weight. Afzal et al. (2008)
NaCl and CaSO4 (50 mM) Increased K+ and Ca2+ concentration;
and decreased Na+ accumulation.
Increased total sugars.
Priming with CaCl2 (50 mg l
1) Increased K+ and decreased Na+ accumulation. Jafar et al. (2012)
enhanced all agronomic and yield attributes
such as plant height, No. tillers, No. spikelets,
No. grains, 1000-grain weight, grain yield,
biological yield and harvest index.
Priming with NaCl (30 mM) Increased root and shoot dry weight Jamal et al. (2011)
increased K+ concentration and K+/Na+
ratio and decreased Na+ accumulation
Canola Priming with NaCl (14 dS m
1) Improved seedling dry weight increased Farhoudi and
accumulation of proline increased K+ Sharifzadeh (2006)
and Ca2+ concentration and
decreased Na+ accumulation
Maize Priming with NaCl, KCl and Increased germination percentage Ashraf and Rauf (2001)
CaCl22H2O (200 meq l
1) and dry weight of seedlings
Melon Priming with NaCl (18 dS m
1) Increased dry weight of seedlings Sivritepe et al. (2003)
enhanced total sugar and proline
accumulation increased K+ and Ca2+
concentration and K+/Na+ ratio and
decreased Na+ accumulation
Mung bean Priming with NaCl (50 mM) Increased growth and photosynthetic Saha et al. (2010)
pigments (Chl a, b and carotenoids)
of seedlings decreased MDA concentration
increased accumulation of proline
Tomato Priming with NaCl and Increased dry weight of seedlings lowered Nawaz et al. (2011)
KNO3 (25 and 50 mM) electrical conductivity of seed leachate
enhanced accumulation of total sugars
Wheat Seedlings exposed to 100 lM SNP Increased amounts of chlorophyll a and b Alavi et al. (2014)
improved MSI (%) and reduced MDA
concentration
Foliar application of NO Enhanced shoot and root fresh weights, Kausar and Shahbaz (2013)
(0.05, 0.1 and 0.15 mM) leaf area and photosynthetic rate
Seedlings treated with Counteract the decay of chlorophyll Ruan et al. (2002)
SNP (0.1 and 1 mM) decreased MDA concentration and
plasma membrane permeability elevated
the concentration of proline
Seed priming with 0.1 mM SNP Increased germination rate and weights Zheng et al. (2009)
of coleoptile and radicle increased total
soluble sugars, K+ concentration and
decreased MDA content and Na+
accumulation of germinating grains
Barley Seedlings treated with 50 lM SNP Decreased ion leakage and MDA Li et al. (2008)
concentration
Chickpea Foliar spray with 0.2 and 1 mM SNP Decreased membrane injury and lipid Sheokand et al. (2010)
peroxidation (MDA concentration)
Cucumber Seed soaking in 50 lM SNP Decreased seedling contents of MDA Fan et al. (2013)
Maize Seedlings treated with 100 lM SNP Increased dry matter accumulation Zhang et al. (2006)
increased chlorophyll content
decreased electrolyte leakage

(continued)

46 © 2015 Blackwell Verlag GmbH, 202 (2016) 37–50


Table 9 (continued)
Plant Treatment
Mangrove Seedlings treated with SNP (100 lM)
(Aegiceras
corniculatum)
Soya bean Seedlings treated with SNP (100 lM)
Tomato Foliar spray with 10 lM SNP
Plants (60 DAS) treated
with SNP (100 lM)
DAS, days after sowing; SNP, sodium nitroprusside.
(Table 9). Under non-saline conditions, the grain yield for
Sakha-93 and Gemmiza-9 was 9.24 and 9.76 t ha
1,
respectively (Abd El-Razek and El-Sheshtawy 2013). Due
to salinity, the grain yields of non-primed plants were
4.747 and 3.840 t ha
1 for Sakha-93 and Gemmiza-9,
respectively (Table 8). Thus, priming treatments, in partic-
ular redox halopriming has the ability to reduce the loss of
grain yield in the wheat genotypes. Our results suggest that
ameliorative effect of redox halopriming on the grain yield
of wheat genotypes was effectively higher in salt-sensitive
genotype than salt-tolerant one.
Conclusion
This study revealed that there is a synergistic effect between
halopriming with diluted sea water (EC = 9 dS m
1) and
nitric oxide (0.2 mM) in alleviating the deleterious effects
of salt stress on growth and yield of wheat genotypes. In
other words, redox halopriming technique has a crucial
role to induce wheat genotypes, particularly salt sensitive,
against salt stress. In addition, this study proved that the
nature of salt tolerance in salt-tolerant genotype may be
related to lower accumulation of Na+ and MDA content, as
well as higher plant biomass, MSI, osmolytes (proline and
TSS) and K+ concentration under salt stress as compared
to the salt-sensitive genotype.
Acknowledgements
We extend our sincere thanks to Dr. Atta Soliman, Depart-
ment of Plant Science, Faculty of Agriculture and Food Sci-
ences, University of Manitoba, Winnipeg, Manitoba, R3T
2N2 Canada, for expeditious revising this manuscript.
© 2015 Blackwell Verlag GmbH, 202 (2016) 37–50
Inducing Salt Tolerance in Bread Wheat
Effects Reference
Increased Chl a, b, total Chl and Chen et al. (2014)
carotenoid concentrations,
as well as Chl a/b ratio decreased
MDA concentration
Increased anthocyanin concentration Simaei et al. (2012)
of leaves decreased LOX activity
of leaves (as indicator of lipid
peroxidation) Improved K+
uptake and inhibited Na+
uptake of roots and shoots
Increased b-carotene and anthocyanin Ali and Ismail (2014)
concentrations in fruits
Improved of shoot and root dry Wu et al. (2011)
weight decreased MDA concentration
increased the concentration of
proline and total soluble sugars
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