Дослідження потенціалу використання молекули основи Шиффа як екзогенного антиоксиданту для насінні ячменю в умовах сольового стресу
Salt stress is one of the most important environmental factors that affect agricultural lands and causes product loss. Today, the application of various natural or synthetic molecules exogenously to plants and efforts to increase plant tolerance against environmental stresses as a result of these ap...
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Plant Introduction| _version_ | 1860145134168965120 |
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| author | Çevik, Sertan |
| author_facet | Çevik, Sertan |
| author_sort | Çevik, Sertan |
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| description | Salt stress is one of the most important environmental factors that affect agricultural lands and causes product loss. Today, the application of various natural or synthetic molecules exogenously to plants and efforts to increase plant tolerance against environmental stresses as a result of these applications have been widely investigated by scientists. In this study, a Schiff base molecule (0, 3, 6, and 9 µM), which has shown in vitro antioxidant properties, was applied to barley seeds under salt stress (0, 50, 150, and 250 mM NaCl). In order to evaluate the effects of this molecule on barley under salt stress, seed germination, growth parameters, lipid peroxidation, proline content, histochemical detection of superoxide and hydrogen peroxide radicals, and mitotic index analysis were conducted. According to the results, salt stress decreased germination parameters, plumule and radicle lengths, and mitotic index while it increased proline content, lipid peroxidation, and radical contents. Schiff base treatment clearly reduced lipid peroxidation and radical content in all groups. However, it also decreased germination and growth parameters and mitotic index. The obtained results showed that the antioxidant property of this molecule was also preserved in plants under stress, but it was also determined that the molecule had negative effects, primarily on cell division. If necessary modifications can be made to the molecule, the negative effects on cell division can be eliminated, and this molecule, which is very easy and cheap to obtain, may be widely used to increase the tolerance of plants against environmental stress. |
| doi_str_mv | 10.46341/PI2022018 |
| first_indexed | 2025-07-17T12:54:10Z |
| format | Article |
| fulltext |
Plant Introduction, 95/96, 85–95 (2022)
© The Authors. This content is provided under CC BY 4.0 license.
RESEARCH ARTICLE
Investigating the using potential of Schiff base molecule as an exogenous
antioxidant on barley seeds under salt stress conditions
Sertan Çevik
Department of Molecular Biology and Genetics, Faculty of Arts and Sciences, Harran University, 63300 Şanlıurfa, Turkey;
srtncvk@gmail.com
Received: 03.09.2022 | Accepted: 27.09.2022 | Published online: 01.10.2022
Abstract
Salt stress is one of the most important environmental factors that affect agricultural lands and causes
product loss. Today, the application of various natural or synthetic molecules exogenously to plants and
efforts to increase plant tolerance against environmental stresses as a result of these applications have
been widely investigated by scientists. In this study, a Schiff base molecule (0, 3, 6, and 9 µM), which has
shown in vitro antioxidant properties, was applied to barley seeds under salt stress (0, 50, 150, and 250 mM
NaCl). In order to evaluate the effects of this molecule on barley under salt stress, seed germination,
growth parameters, lipid peroxidation, proline content, histochemical detection of superoxide and
hydrogen peroxide radicals, and mitotic index analysis were conducted. According to the results,
salt stress decreased germination parameters, plumule and radicle lengths, and mitotic index while it
increased proline content, lipid peroxidation, and radical contents. Schiff base treatment clearly reduced
lipid peroxidation and radical content in all groups. However, it also decreased germination and growth
parameters and mitotic index. The obtained results showed that the antioxidant property of this molecule
was also preserved in plants under stress, but it was also determined that the molecule had negative
effects, primarily on cell division. If necessary modifications can be made to the molecule, the negative
effects on cell division can be eliminated, and this molecule, which is very easy and cheap to obtain, may
be widely used to increase the tolerance of plants against environmental stress.
Keywords: barley, Schiff base, reactive oxygen species, salt stress
https://doi.org/10.46341/PI2022018
UDC 581.19
Funding: The author declares no funding.
Competing Interests: The author declares no conflict of interest.
Introduction
Plants are exposed to various biotic and
abiotic environmental stressors throughout
their lives. These stresses negatively affect all
metabolic events in plants and cause serious
crop losses (Raza et al., 2019). Salinity, one
of the most important abiotic factors, is one
of the most restrictive stress factors for
agricultural production today (Güzel Değer
& Çevik, 2021). In addition to ion toxicity,
low osmotic potential occurs in the soil,
which makes it difficult for the plant to take
water from the soil (Ahmad & Akhtar, 2019).
Studies show that almost 20 % of arable land
is constantly exposed to salt stress. However,
many studies have been conducted worldwide
to understand the mechanism of salt stress
tolerance and the responses given by plants,
https://creativecommons.org/licenses/by/4.0/
https://orcid.org/0000-0003-1259-7863
86 Plant Introduction • 95/96
Sertan Çevik
but little progress has been made in this
regard. The most important reason for this
is the complex physiological and genetic
mechanism of salt stress tolerance and the
lack of reliable screening methods (Zhu et al.,
2020).
Environmental stresses (abiotic and biotic),
which are common today due to severe and
harsh climate change (Hasanuzzaman et al.,
2020), can trigger oxidative stress in plants,
causing the formation of reactive oxygen
species (ROS). ROS are partially reduced or
activated oxygen derivatives containing both
free radical and non-radical forms that cause
cellular damage and metabolic disorders (Jaleel
et al., 2009). Plants cope with oxidative stress
through an endogenous defense mechanism
consisting of enzymatic and non-enzymatic
antioxidants (Kaur et al., 2019).
Recent studies have shown that ROS are
not entirely harmful and are necessary for the
stabilization of the intracellular redox state at
low concentrations (Circu & Aw, 2010; Schieber
& Chandel, 2014). The ROS level is maintained
by the balance between ROS generation and
ROS scavenging. However, during stress
conditions, excessive ROS production upsets
the balance and causes cellular damage,
reducing plant productivity (Hasanuzzaman
et al., 2020). In order to minimize product
loss due to environmental stress, researchers
generally adopted two different approaches.
One of them is to develop varieties resistant
to environmental stresses through traditional
or modern methods, and the other is to
strengthen the antioxidant systems of plants
by exogenous applications (Özkoku et al.,
2019). Developing cultivars resistant to
environmental stresses through traditional or
modern techniques is time-consuming and
quite difficult (Breseghello & Coelho, 2013).
These situations have led researchers to make
external applications that strengthen the
antioxidant system, which is a more practical
way. When the studies conducted in this
context are examined, it is seen that synthetic
or natural molecules are applied to plants in
an extensive scope, especially under stress
conditions (Çevik et al., 2014, 2019; Bekfelavi
et al., 2021).
Schiff bases are important functional
groups due to their wide biological and
chemical functions. In addition to their broad
chemical properties, the biological functions
of Schiff bases, such as antibacterial,
antifungal, antioxidant, antitumor, and
antiviral action, have been reported by
various researchers (Yılmaz, 2021). In
this study, a Schiff base molecule, whose
antioxidant properties were demonstrated
by in vitro methods by Yılmaz (2021), was
applied to barley seeds under salt stress, and
the effects of this molecule were investigated
with extensive morphological, physiological,
and biochemical analyses. According to the
literature, this is the first study to examine
the effects of a Schiff base molecule,
which has been shown to have antioxidant
properties in vitro by applying to a plant
under salt stress. The obtained data provided
important information for the future use of
such molecules.
Material and methods
Plant material and treatments
Hordeum vulgare L. seeds were obtained from
ALATA Horticultural Research Station. Seeds
were kept in 5% sodium hypochlorite for
5 min and washed five times with pure water
for surface sterilization prior to use. Schiff
base molecule (8e) (Fig. 1), which has high
in vitro DPPH free radical scavenging activity,
has been used in this study. Details on the
synthesis steps of this molecule are given by
Yılmaz (2021).
A preliminary experiment was performed
to determine the optimum concentration of
8e molecules for barley seeds according to
germination data. Solutions containing Schiff
base molecule (0, 3, 6, and 9 µM of 8e) and salt
(0, 50, 150, and 250 mM of NaCl) were added
to Petri dishes in equal amounts, and water
N OH
OH
Figure 1. The Schiff base molecule (8e) applied in
this study.
Plant Introduction • 95/96 87
Schiff base molecule as an exogenous antioxidant on barley seeds under salt stress
Groups NaCl
(mM)
8e
(µM)
Groups NaCl
(mM)
8e
(µM)
Groups NaCl
(mM)
8e
(µM)
Groups NaCl
(mM)
8e
(µM)
1 0 0 5 0 3 9 0 6 13 0 9
2 50 0 6 50 3 10 50 6 14 50 9
3 150 0 7 150 3 11 150 6 15 150 9
4 250 0 8 250 3 12 250 6 16 250 9
Table 1. Experimental application groups.
was used as a control group. Ten seeds were
placed in each Petri dish, and ten Petri dishes
were used for each application group. Applied
combinations are shown in Table 1.
Seed germination and seedling growth
parameters
Seeds were imbibed in aerated water for one
day at 22 °C and then transferred to Petri
dishes. Seeds in Petri dishes were germinated
at 24 : 18 °C day: night temperature, 16 : 8
day: night light period, 150 μmol m-2s-1 light
intensity, and 60 ± 5 % humidity conditions for
three days under controlled conditions in the
climate room. The radicle and plumule lengths
were measured with a digital caliper at the end
of the third day.
Seeds were considered to have germinated
when the radicles were ≥2 mm long. The
number of germinated seeds was recorded
daily, and the final germination percentage
was determined after three days. Germination
rate (M) was calculated according to the
formula M = n1 / d1 + n2 / d2+ n3 / d3, where
n is the number of germinated seeds, and
d is a day (Ranal & Santana, 2006). Mean
germination time (MGT) was calculated by
using the equation MGT = ∑ (n × d) / N, where
n is the number of seeds germinated on
each day, d is the number of days, and N is
the total number of germinated seeds (Ellis &
Roberts, 1981). Seed germination percentage
(SG) was calculated using the following
formula SG [%] = Number of germinated
seeds / Total number of seeds × 100 (Czabator,
1962). Germination index (GI) was calculated
according to the formula GI = ∑ (ni × Ti) / N,
where n is the number of newly germinating
seeds, N is the total number of seeds; and ni
is the number of seeds germinated at day Ti
(Aravind et al., 2019).
Lipid peroxidation
Lipid peroxidation was determined by
measuring the malondialdehyde (MDA) content
according to Ohkawa et al. (1979). Radicle
and plumule tissue (0.2 g) was homogenized
1 mL (5 %) trichloroacetic acid (TCA) solution.
The homogenate was centrifuged for 10 min
at 8,000 rpm. After that, supernatant,
thiobarbituric acid, and TCA solutions were
mixed in equal volumes in tubes, and tubes
were incubated at 96 °C for 25 min. The tubes
were placed in an ice bath to terminate the
reaction and centrifuged at 6,000 rpm for
5 min. The mixture was measured at 532 and
600 nm (Shimadzu 1800 240V). MDA content
was calculated using the extinction coefficient
of 155 mM-1cm-1.
Free proline content
Free proline content was determined
according to Bates et al. (1973). The radicle and
plumule samples (0.25 g) were homogenized
in 3 % sulfosalicylic acid. The homogenate
was centrifuged for 3 min at 3,000 rpm,
and then the supernatant was mixed well
with acid ninhydrin and glacial acetic acid
in equal volumes and incubated at 100 °C
for 60 min. The reaction was terminated by
adding cold toluene (4 mL) to the tubes. The
toluene phase was evaporated and analyzed by
spectrophotometry (Shimadzu 1800 240V) at
520 nm. Proline concentration was calculated
by using a calibration curve and expressed as
µmol proline g-1 FW.
Detection of superoxide and hydrogen
peroxide radicals in barley roots
Superoxide radicals were detected following
Piacentini et al. (2021). In this method, NBT is
used to monitor the intracellular production
of the superoxide anion. Roots were exposed
88 Plant Introduction • 95/96
Sertan Çevik
to NBT solution (0.5 mg / mL NBT in 10 mM
Tris-HCl, pH 7.4) for 30 min. NBT is reduced
by superoxide radicals, and blue color forms
on the root surface as superoxide radicals
reduce NBT.
H2O2 was detected by 3,3′-diamino-
benzidine (DAB) staining method according
to Thordal-Christensen et al. (1997) with
minor modifications. Briefly, barley roots
were treated with DAB staining buffer for
60 min (in the dark), and the reaction was
stopped by adding ethanol:glycerol:acetic
acid (3 : 1 : 1, v/v). The oxidized DAB formed
a brown precipitate on the surface of the
roots and was visualized by light microscopy
(Olympus BX53).
Mitotic index analysis
For mitotic index analysis, 1–1.5 cm barley
root tips were cutted and fixed for 24 h in
ethanol : glacial acetic acid (3 : 1, v/v), then
stored in 70 % ethanol at +4 °C until analysis.
The root tips were hydrolyzed in 2M HCl for
20 min at 60 °C and stained by the Feulgen
method (Tabur & Demir, 2010). The slides
were examined under an optical microscope
(Olympus BX53). A mean of 1000 cells was
counted from each root to get a total of 5,000
cells per treatment. The mitotic index (MI)
was determined using the formula: MI [%]= the
number of dividing cells/the number of totally
examined cells × 100.
Experimental design and statistical analysis
Salt stress and Schiff base treatment were
carried out according to a completely
randomized experimental design with two
factors. Treatments had three replications
for proline, MDA, and radical detection
analysis; five replications for germination
%, germination rate, germination index,
mean germination time, and mitotic index
analysis; ten replications for plumule and
radicle length analysis. All quantitative data
expressed as percentages were subjected to
arcsine transformation. Data were subjected
to analysis of variance (ANOVA), and the
means were separated using the Least
Significant Difference (LSD) multiple range
test at p ≤ 0.05. All the statistical analyses
were performed using the JMP ver. 8
(SAS Institute Inc.) software package. The
coefficients of variation were shown in the
tables to demonstrate the reliability of the
experiment.
Results and discussion
Salt stress affects plants and causes serious
production problems in the field. The
results of this study demonstrate that
with an increasing degree of salt stress,
germination percentage, germination rate,
and germination index parameters of barley
decreased, while mean germination time
(MGT) increased (Table 2). Other researchers
also reported similar results (Yildirim &
Güvenc, 2006; Dehnavi et al., 2020). Salt
stress can directly inhibit germination
parameters by making difficult water uptake
for seeds, or it can reduce germination rates
due to ionic toxicity (Yildirim & Güvenc,
2006). Exogenous treatment by 8e increased
MGT under control conditions and mild salt
stress while decreased MGT under severe salt
stress conditions. These interesting findings
show that exogenous Schiff base application
can reduce germination time, especially
under severe stress conditions. A low MGT
value indicates faster germination compared
to a high MGT value (Yongkriat et al., 2020).
The fact that MGT decreases under stress by
Schiff base treatment may be an important
advantage for plants to cope with salt stress.
As seen from Table 3, radicle and plumule
lengths decreased due to increased salt
concentration. Other researchers also reported
similar results on different plants (Keshavarzi,
2011; El-Bastawisy et al., 2018). Generally,
exogenous 8e applications decreased the
radicle and plumule lengths. This result was
also supported by mitotic index (MI) analysis.
Especially under control conditions increasing
8e concentration decreased MI, but under
severe salt stress conditions, 8e did not affect
MI. These results are in good agreement
with the radicle length results in this study.
These findings present an important area
to be investigated for the use of Schiff base
molecules, which have antioxidant properties
in vitro, before such exogenous applications.
If the reasons for the negative effects of these
molecules on cell division can be found, the
obstacles to their use will be overcome with
the necessary molecular modifications.
Plant Introduction • 95/96 89
Schiff base molecule as an exogenous antioxidant on barley seeds under salt stress
Treatment Seed germination
percentage (%)
Germination rate Germination ındex Mean germination
time (days)
Control 92.60 ± 1.02 a 63.54 ± 1.12 a 2.06 ± 0.03 a 1.76 ± 0.02 j
0 NaCl : 3 µM 8e 89.00 ± 1.10 b 54.07 ± 0.57 b 1.81 ± 0.03 b 1.97 ± 0.02 gh
0 NaCl : 6 µM 8e 88.00 ± 2.00 b 51.04 ± 0.89 c 1.74 ± 0.03 c 2.02 ± 0.03 f
0 NaCl : 9 µM 8e 86.20 ± 0.75 c 49.64 ± 0.83 d 1.67 ± 0.02 d 2.06 ± 0.02 e
50 mM NaCl : 0 8e 80.80 ± 2.14 d 50.38 ± 0.53 cd 1.67 ± 0.01 d 1.94 ± 0.02 h
50 mM NaCl : 3 µM 8e 79.20 ± 1.60 d 53.16 ± 0.71 b 1.69 ± 0.02 d 1.87 ± 0.02 i
50 mM NaCl : 6 µM 8e 76.40 ± 1.02 e 39.92 ± 0.56 f 1.44 ± 0.03 e 2.11 ± 0.03 d
50 mM NaCl : 9 µM 8e 74.80 ± 1.33 e 44.14 ± 0.45 e 1.43 ± 0.01 e 2.09 ± 0.03 de
150 mM NaCl : 0 8e 57.20 ± 0.75 g 40.18 ± 0.35 f 1.28 ± 0.02 f 2.23 ± 0.02 b
150 mM NaCl : 3 µM 8e 64.20 ± 0.98 f 36.29 ± 0.36 g 1.18 ± 0.02 g 2.00 ± 0.02 f
150 mM NaCl : 6 µM 8e 48.00 ± 2.28 h 27.31 ± 0.43 h 0.87 ± 0.04 h 1.96 ± 0.03 gh
150 mM NaCl : 9 µM 8e 41.00 ± 1.10 i 21.78 ± 0.33 i 0.74 ± 0.03 i 1.99 ± 0.03 fg
250 mM NaCl : 0 8e 32.80 ± 1.60 j 18.18 ± 0.61 j 0.58 ± 0.02 j 2.24 ± 0.01 ab
250 mM NaCl : 3 µM 8e 26.20 ± 1.17 k 15.18 ± 0.48 l 0.49 ± 0.03 k 1.96 ± 0.02 gh
250 mM NaCl : 6 µM 8e 31.40 ± 1.02 j 16.17 ± 0.42 k 0.54 ± 0.03 l 2.26 ± 0.02 a
250 mM NaCl : 9 µM 8e 26.40 ± 1.50 k 13.34 ± 0.44 m 0.44 ± 0.01 m 2.19 ± 0.02 c
LSD, p < 0.001 1.428 0.534 0.034 0.032
Table 2. Effect of salt stress and 8e treatment on germination of barley. Different letters indicate statistically
significant differences between groups.
Treatment Plumule length (mm) Radicle length (mm) Mitotic ındex
Control 6.07 ± 0.70 a 6.78 ± 0.59 a 0.154 ± 0.005 a
0 NaCl : 3 µM 8e 4.73 ± 0.75 b 5.53 ± 0.25 b 0.123 ± 0.004 b
0 NaCl : 6 µM 8e 4.41 ± 0.63 bc 5.45 ± 0.58 b 0.121 ± 0.002 b
0 NaCl : 9 µM 8e 4.21 ± 0.72 cd 4.32 ± 0.34 c 0.094 ± 0.004 c
50 mM NaCl : 0 8e 3.94 ± 0.63 d 5.21 ± 0.61 b 0.116 ± 0.002 b
50 mM NaCl : 3 µM 8e 2.86 ± 0.41 e 4.48 ± 0.31 c 0.097 ± 0.002 d
50 mM NaCl : 6 µM 8e 2.21 ± 0.41 f 4.31 ± 0.70 c 0.093 ± 0.003 d
50 mM NaCl : 9 µM 8e 1.93 ± 0.21 fg 3.62 ± 0.51 d 0.073 ± 0.003 e
150 mM NaCl : 0 8e 2.03 ± 0.27 f 3.46 ± 0.82 de 0.072 ± 0.003 fg
150 mM NaCl : 3 µM 8e 1.52 ± 0.49 gh 3.33 ± 0.29 de 0.069 ± 0.003 fg
150 mM NaCl : 6 µM 8e 1.48 ± 0.34 h 3.13 ± 0.50 e 0.073 ± 0.004 fg
150 mM NaCl : 9 µM 8e 1.47 ± 0.34 h 3.40 ± 0.19 de 0.070 ± 0.001 fg
250 mM NaCl : 0 8e 0.71 ± 0.34 i 1.43 ± 0.31 f 0.032 ± 0.001 h
250 mM NaCl : 3 µM 8e 0.13 ± 0.05 j 1.31 ± 0.27 fg 0.030 ± 0.002 hl
250 mM NaCl : 6 µM 8e 0.13 ± 0.05 j 1.57 ± 0.24 f 0.036 ± 0.002 h
250 mM NaCl : 9 µM 8e 0.10 ± 0.01 j 1.07 ± 0.40 g 0.024 ± 0.002 l
LSD, p < 0.001 0.439 0.436 0.244
Table 3. Effect of salt stress and 8e treatment on growth of barley.. Different letters indicate statistically
significant differences between groups.
90 Plant Introduction • 95/96
Sertan Çevik
Figure 2. Distribution of hydrogen peroxide in barley roots visualized by 3,3′-diaminobenzidine staining.
Dark-stained regions indicate hydrogen peroxide produced in cells.
Figure 3. Distribution of superoxide radicals in barley roots visualized by nitroblue tetrazolium staining.
Dark-stained regions indicate superoxide produced in cells.
Plant Introduction • 95/96 91
Schiff base molecule as an exogenous antioxidant on barley seeds under salt stress
Salt stress increased the content of
hydrogen peroxide (Fig. 2) and superoxide
radical (Fig. 3) in the barley roots compared
to control groups. It was observed that
there is a strong correlation between the
radical content and the increase in the
salt concentration. Exogenous application
of 8e decreased both superoxide and
hydrogen peroxide radical contents. The
in vitro antioxidant properties of this
molecule were demonstrated by Yılmaz
(2021). In this study, the fact that the
exogenous application reduces the internal
radical content shows that the antioxidant
properties of the molecule are also
preserved in-plant applications. However,
as was emphasized before, the negative
effect of the exogenous application of
this molecule on cell division reduces
its potential for use as an exogenous
antioxidant. Therefore, if a specific cause
affecting cell division of this molecule
is found, it may be used as an exogenous
antioxidant in such conditions.
Salt stress increased proline content in
barley roots. Exogenous Schiff base treatment
also increased proline content under severe
salt stress conditions (Fig. 4). It has been
demonstrated in different studies that the
amount of proline increases under many
environmental stresses (Vendruscolo et al.,
2007; Lum et al., 2014; Chun & Chandrasekaran,
2018; Çevik et al., 2019). The best-known
feature of proline is that it is a good osmotic
preservative. In addition to being a good
osmotic protector, it has been reported in
recent years that proline reduces radical
formation and shows antioxidant properties
(Szabados & Savoure, 2010). Increasing proline
content by Schiff base application may be
important for plants under stress conditions.
However, how exogenous Schiff base
application increases the amount of proline
under stress is another important question.
The fact that this increase occurred only
under severe salt stress conditions should also
be investigated.
The MDA content, which increased
with salt stress, decreased significantly
by the exogenous Schiff base treatment
(Fig. 5). MDA is the end product of lipid
peroxidation and a good marker of
oxidative damage. With the increase in
the amount of intracellular radicals, lipid
peroxidation occurs (Gawel et al., 2004).
In this study, the fact that the Schiff base
application decreased the content of H2O2
and superoxide radicals may have caused
a decrease in MDA content. Protecting
cell membranes under stressful conditions
Figure 4. Effect of salt stress and 8e treatment on proline content of barley. LDS = 0.428, p < 0.001. Different
letters indicate statistically significant differences between groups.
92 Plant Introduction • 95/96
Sertan Çevik
gives the plant a significant advantage in
coping with stress. Reducing MDA content
by exogenous 8e treatment is important in
plant stress tolerance.
Conclusions
Salt stress, which is one of the most
important environmental factors, is a
chronic stress. The difficulty of remediation
of saline soils necessitates increasing plant
tolerance to this stress. However, approaches
increasing salt tolerance by exogenous
applications have recently become very
popular due to genetic constraints and
insufficient time. In this study, a Schiff
base molecule, whose in vitro antioxidant
properties were shown in another study, was
exogenously applied to barley plants under
salt stress for the first time. According to
the results, the application of this molecule
decreased the amount of radicals and MDA
that increased by salt stress. However, it
was determined that the mitotic index
and the growth parameters were also
negatively affected by this application. The
findings revealed that to avoid the negative
effect, Schiff base molecules, which have
antioxidant properties, need to be modified
before application in a salt stress regulation.
If the factors affecting cell division can be
eliminated, these molecules, which are easy
and cheap to obtain, may be widely applied
to modify plant salt tolerance.
Acknowledgements
I thank Dr. Sara Yasemin (Siirt University) for
her contributions to statistical analyses and
Dr. Özgür Yılmaz (Mersin University) for Schiff
base molecule supply.
References
Ahmad, I., & Akhtar, M.S. (2019). Use of
nanoparticles in alleviating salt stress. In
M.S. Akhtar (Ed.), Salt stress, microbes, and
plant interactions: causes and solution (pp. 199–
215). Springer, Singapore. https://doi.
org/10.1007/978-981-13-8801-9_9
Aravind, J., Vimala, D., Radharani, J., Jacob, S.,
& Srinivasa, K. (2019). The germination metrics
package: a brief introduction. ICAR-National
Bureau of Plant Genetic Resources, New Delhi,
India.
Bates, L.S., Waldren, R.P., & Teare, I.D. (1973).
Rapid determination of free proline for water-
stress studies. Plant and Soil, 39(1), 205–207.
https://doi.org/10.1007/BF00018060
Figure 5. Effect of salt stress and 8e treatment on MDA content of barley. LDS = 1.066, p < 0.001. Different
letters indicate statistically significant differences between groups.
https://doi.org/10.1007/978-981-13-8801-9_9
https://doi.org/10.1007/978-981-13-8801-9_9
https://doi.org/10.1007/BF00018060
Plant Introduction • 95/96 93
Schiff base molecule as an exogenous antioxidant on barley seeds under salt stress
Bekfelavi, E.Y., Yildizli, A., Kuş, N.Ş., Çevik. S.,
& Ünyayar, S. (2021). Osmoprotectant and
antioxidant effects of new synthesized
6-(2-hydroxyethyl)cyclohex-3-enol on barley
under drought stress. Biologia Futura, 72(2),
241–249. https://doi.org/10.1007/s42977-020-
00058-w
Breseghello, F., & Coelho, A.S.G. (2013). Traditional
and modern plant breeding methods with
examples in rice (Oryza sativa L.). Journal of
Agricultural and Food Chemistry, 61(35), 8277–
8286. https://doi.org/10.1021/jf305531j
Çevik, S., Güzel Değer, A., Yıldızlı, A., Doğanyiğit, N.,
Gök, A., & Unyayar, S. (2019). Proteomic and
physiological analyses of dl-cyclopentane-1,2,3-
triol-treated barley under drought stress. Plant
Molecular Biology Reporter, 37, 237–251. https://
doi.org/10.1007/s11105-019-01151-8
Çevik, S., Yıldızlı, A., Yandım, G., Göksu, H.,
Gultekin, M.S., Güzel Değer, A., Çelik, A.,
Şimşek Kuş, N., & Ünyayar, S. (2014). Some
synthetic cyclitol derivatives alleviate the effect
of water deficit in cultivated and wild-type
chickpea species. Journal of Plant Physiology,
171(10), 807–816. https://doi.org/10.1016/j.
jplph.2014.01.010
Chun, S.C., & Chandrasekaran, M. (2018). Proline
accumulation influenced by osmotic stress
in arbuscular mycorrhizal symbiotic plants.
Frontiers in Microbiology, 9, Article 2525. https://
doi.org/10.3389/fmicb.2018.02525
Circu, M.L., & Aw, T.Y. (2010). Reactive oxygen
species, cellular redox systems, and
apoptosis. Free Radical Biology and Medicine,
48(6), 749–762. https://doi.org/10.1016/j.
freeradbiomed.2009.12.022
Czabator, F.J. (1962). Germination value: an index
combining speed and completeness of pine
seed germination. Forest Science, 8, 386–395.
https://academic.oup.com/forestscience/article-
abstract/8/4/386/4746567
Dehnavi, R.A., Zahedi, M., Ludwiczak, A., Cardenas
Perez, S., & Piernik, A. (2020). Effect of salinity
on seed germination and seedling development
of sorghum (Sorghum bicolor (L.) Moench)
genotypes. Agronomy, 10(6), Article 859. https://
doi.org/10.3390/agronomy10060859
El-Bastawisy, Z.M., El-Katony, T.M., & Abd El-
Fatah, S.N. (2018). Genotypic variability in salt
tolerance of Vicia faba during germination
and early seedling growth. Journal of King Saud
University-Science, 30(2), 270–277. https://doi.
org/10.1016/j.jksus.2017.04.004
Ellis, R.H., & Roberts, E.H. (1981). The quantification
of ageing and survival in orthodox seeds. Seed
Science and Technology, 9, 373–409.
Gawel, S., Wardas, M., Niedworok, E., & Wardas, P.
(2004). Malondialdehyde (MDA) as a lipid
peroxidation marker. Wiadomości Lekarskie, 57(9–
10), 453–455.
Güzel Değer, A.G., & Çevik, S. (2021). Impacts of
nano-TiO2 on the initial development stages of
barley seedlings under salinity. Mediterranean
Agricultural Sciences, 34(1), 109-116. https://doi.
org/10.29136/mediterranean.816107
Hasanuzzaman, M., Bhuyan, M.H.M., Zulfiqar, F.,
Raza, A., Mohsin, S.M., Mahmud, J.A., &
Fotopoulos, V. (2020). Reactive oxygen species
and antioxidant defense in plants under abiotic
stress: revisiting the crucial role of a universal
defense regulator. Antioxidants, 9(8), Article 681.
https://doi.org/10.3390/antiox9080681
Jaleel, C.A., Manivannan, P., Wahid, A., Farooq, M.,
Al-Juburi, H.J., Somasundaram, R., &
Panneerselvam, R. (2009). Drought stress in
plants: a review on morphological characteristics
and pigments composition. International Journal
of Agriculture and Biology, 11(1), 100–105.
Kaur, N., Kaur, J., Grewal, S.K., & Singh, I.
(2019). Efect of heat stress on antioxidative
defense system and its amelioration by heat
acclimation and salicylic acid pre-treatments
in three pigeonpea genotypes. Indian Journal of
Agricultural Biochemistry, 32(1), 106–110. https://
doi.org/10.5958/0974-4479.2019.00014.5
Keshavarzi, M.H.B. (2011). Effect of salt stress on
germination and early seedling growth of savory
(Satureja hortensis). Australian Journal of Basic and
Applied Sciences, 5(12), 3274–3279.
Lum, M.S., Hanafi, M.M., Rafii, Y.M., & Akmar, A.S.N.
(2014). Effect of drought stress on growth,
proline and antioxidant enzyme activities of
upland rice. Journal of Animal and Plant Sciences,
24(4), 1487–1493.
Ohkawa, H., Ohishi, N., & Yagi, K. (1979). Assay for
lipid peroxides in animal tissues by thiobarbituric
acid reaction. Analytical Biochemistry, 95(2),
351–358. https://doi.org/10.1016/0003-
2697(79)90738-3
Özkoku, G., Cevik, S., & Ünyayar, S. (2019). The
effect of exogenous application of synthetic
inositol derivative (allo-inositol) on salt tolerance
of Capsicum chinense. Anadolu Tarim Bilimleri
Dergisi, 34(3), 319–326. https://doi.org/10.7161/
omuanajas.556100
Piacentini, D., Falasca, G., Canepari, S., &
Massimi, L. (2019). Potential of PM-selected
components to induce oxidative stress and
root system alteration in a plant model
organism. Environment International, 132,
Article 105094. https://doi.org/10.1016/j.
envint.2019.105094
https://doi.org/10.1007/s42977-020-00058-w
https://doi.org/10.1007/s42977-020-00058-w
https://doi.org/10.1021/jf305531j
https://doi.org/10.1007/s11105-019-01151-8
https://doi.org/10.1007/s11105-019-01151-8
https://doi.org/10.1016/j.jplph.2014.01.010
https://doi.org/10.1016/j.jplph.2014.01.010
https://doi.org/10.3389/fmicb.2018.02525
https://doi.org/10.3389/fmicb.2018.02525
https://doi.org/10.1016/j.freeradbiomed.2009.12.022
https://doi.org/10.1016/j.freeradbiomed.2009.12.022
https://academic.oup.com/forestscience/article-abstract/8/4/386/4746567
https://academic.oup.com/forestscience/article-abstract/8/4/386/4746567
https://doi.org/10.3390/agronomy10060859
https://doi.org/10.3390/agronomy10060859
https://doi.org/10.1016/j.jksus.2017.04.004
https://doi.org/10.1016/j.jksus.2017.04.004
https://doi.org/10.29136/mediterranean.816107
https://doi.org/10.29136/mediterranean.816107
https://doi.org/10.3390/antiox9080681
https://doi.org/10.5958/0974-4479.2019.00014.5
https://doi.org/10.5958/0974-4479.2019.00014.5
https://doi.org/10.1016/0003-2697(79)90738-3
https://doi.org/10.1016/0003-2697(79)90738-3
https://doi.org/10.7161/omuanajas.556100
https://doi.org/10.7161/omuanajas.556100
https://doi.org/10.1016/j.envint.2019.105094
https://doi.org/10.1016/j.envint.2019.105094
94 Plant Introduction • 95/96
Sertan Çevik
Дослідження потенціалу використання молекули основи Шиффа як екзогенного
антиоксиданту для насінні ячменю в умовах сольового стресу
Сертан Чевік
Кафедра молекулярної біології та генетики, факультет мистецтв і наук, Харранський університет,
63300 Шанлиурфа, Туреччина; srtncvk@gmail.com
Сольовий стрес є одним із найважливіших факторів навколишнього середовища, який впливає на
сільськогосподарські угіддя та спричиняє втрати продукції. Сьогодні вчені широко досліджують
екзогенне застосування різних природних або синтетичних молекул щодо рослин і намагаються
підвищити стійкість рослин до стресів навколишнього середовища. У цьому дослідженні молекула
основи Шиффа (0, 3, 6 та 9 µM), яка показала антиоксидантні властивості in vitro, була застосована до
насіння ячменю під сольовим стресом (0, 50, 150 та 250 mM NaCl). Для того, щоб оцінити вплив цієї
молекули на ячмінь в умовах сольового стресу, було досліджено схожість насіння, параметри росту,
перекисне окислення ліпідів, вміст проліну, а також гістохімічне виявлення радикалів супероксиду та
пероксиду водню та аналіз мітотичного індексу. Згідно з результатами, сольовий стрес зменшував
параметри проростання, довжину брунечок та корінців, а також мітотичний індекс, водночас
збільшував вміст проліну, перекисне окислення ліпідів та вміст радикалів. Обробка основою
Шиффа явно зменшила перекисне окислення ліпідів і вміст радикалів у всіх дослідних групах.
Однак водночас це знизило параметри проростання і росту, а також мітотичний індекс. Отримані
результати показали, що антиоксидантна властивість цієї молекули збереглася в рослинах під час
стресу, але також було визначено, що молекула негативно впливає, насамперед на поділ клітин.
Ranal, M.A., & Santana, D.G. (2006). How and
why to measure the germination process?
Brazilian Journal of Botany, 29(1), 1–11. https://doi.
org/10.1590/S0100-84042006000100002
Raza, A., Razzaq, A., Mehmood, S.S., Zou, X.,
Zhang, X., Lv, Y., & Xu, J. (2019). Impact of climate
change on crops adaptation and strategies to
tackle its outcome: a review. Plants, 8(2), Article
34. https://doi.org/10.3390/plants8020034
Schieber, M., & Chandel, N.S. (2014). ROS
function in redox signaling and oxidative stress.
Current Biology, 24(10), R453–R462. https://doi.
org/10.1016/j.cub.2014.03.034
Szabados, L., & Savoure, A. (2010). Proline: a
multifunctional amino acid. Trends in Plant
Science, 15(2), 89–97. https://doi.org/10.1016/j.
tplants.2009.11.009
Tabur, S., & Demir, K. (2010). Role of some growth
regulators on cytogenetic activity of barley under
salt stress. Plant Growth Regulation, 60(2), 99–104.
https://doi.org/10.1007/s10725-009-9424-6
Thordal-Christensen, H., Zhang, Z., Wei, Y., &
Collinge, D.B. (1997). Subcellular localization of
H2O2 in plants. H2O2 accumulation in papillae
and hypersensitive response during the barley-
powdery mildew interaction. The Plant Journal,
11, 1187–1194. https://doi.org/10.1046/j.1365-
313X.1997.11061187.x
Vendruscolo, E.C.G., Schuster, I., Pileggi, M.,
Scapimd, C.A., Molinarie, H.B.C., Marure, C.J.,
& Vieira, L.G.E. (2007). Stress-induced synthesis
of proline confers tolerance to water deficit
in transgenic wheat. Journal of Plant Physiology,
164(10), 1367–1376. https://doi.org/10.1016/j.
jplph.2007.05.001
Yildirim, E., & Güvenc, İ. (2006). Salt tolerance
of pepper cultivars during germination and
seedling growth. Turkish Journal of Agriculture and
Forestry, 30(5), 347–353.
Yılmaz, Ö. (2021). Synthesis of new Schiff bases;
investigation of their in situ catalytic activity for
Suzuki C-C coupling reactions and antioxidant
activities. Journal of the Chinese Chemical Society, 68(5),
917–928. https://doi.org/10.1002/jccs.202000433
Yongkriat, K.O., Leksungnoen, N., Onwimon, D.,
& Doomnil, P. (2020). Germination and salinity
tolerance of seeds of sixteen Fabaceae species
in Thailand for reclamation of salt-affected lands.
Biodiversitas – Journal of Biological Diversity, 21(5), 215–
222. https://doi.org/10.13057/biodiv/d210547
Zhu, J., Fan, Y., Shabala, S., Li, C., Lv, C., Guo, B., &
Zhou, M. (2020). Understanding mechanisms
of salinity tolerance in barley by proteomic and
biochemical analysis of near-isogenic lines.
International Journal of Molecular Sciences, 21(4), Article
1516. https://doi.org/10.3390/ijms21041516
https://doi.org/10.1590/S0100-84042006000100002
https://doi.org/10.1590/S0100-84042006000100002
https://doi.org/10.3390/plants8020034
https://doi.org/10.1016/j.cub.2014.03.034
https://doi.org/10.1016/j.cub.2014.03.034
https://doi.org/10.1016/j.tplants.2009.11.009
https://doi.org/10.1016/j.tplants.2009.11.009
https://doi.org/10.1007/s10725-009-9424-6
https://doi.org/10.1046/j.1365-313X.1997.11061187.x
https://doi.org/10.1046/j.1365-313X.1997.11061187.x
https://doi.org/10.1016/j.jplph.2007.05.001
https://doi.org/10.1016/j.jplph.2007.05.001
https://doi.org/10.1002/jccs.202000433
https://doi.org/10.13057/biodiv/d210547
https://doi.org/10.3390/ijms21041516
Plant Introduction • 95/96 95
Schiff base molecule as an exogenous antioxidant on barley seeds under salt stress
Якщо в молекулу було б можливо внести необхідні модифікації, можна було б усунути її негативний
вплив на поділ клітин, і тоді цю молекулу, яку дуже легко і дешево отримати, можна було би широко
використовувати для підвищення стійкості рослин до стресу навколишнього середовища.
Ключові слова: ячмінь, основа Шиффа, активні форми кисню, сольовий стрес
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| id | oai:ojs2.plantintroduction.org:article-1618 |
| institution | Plant Introduction |
| keywords_txt_mv | keywords |
| language | English |
| last_indexed | 2025-07-17T12:54:10Z |
| publishDate | 2022 |
| publisher | M.M. Gryshko National Botanical Garden of the NAS of Ukraine |
| record_format | ojs |
| resource_txt_mv | wwwplantintroductionorg/88/5799076b231f04e300dcbb159e0b7788.pdf |
| spelling | oai:ojs2.plantintroduction.org:article-16182023-08-26T20:38:45Z Investigating the using potential of Schiff base molecule as an exogenous antioxidant on barley seeds under salt stress conditions Дослідження потенціалу використання молекули основи Шиффа як екзогенного антиоксиданту для насінні ячменю в умовах сольового стресу Çevik, Sertan Salt stress is one of the most important environmental factors that affect agricultural lands and causes product loss. Today, the application of various natural or synthetic molecules exogenously to plants and efforts to increase plant tolerance against environmental stresses as a result of these applications have been widely investigated by scientists. In this study, a Schiff base molecule (0, 3, 6, and 9 µM), which has shown in&nbsp;vitro antioxidant properties, was applied to barley seeds under salt stress (0, 50, 150, and 250&nbsp;mM NaCl). In order to evaluate the effects of this molecule on barley under salt stress, seed germination, growth parameters, lipid peroxidation, proline content, histochemical detection of superoxide and hydrogen peroxide radicals, and mitotic index analysis were conducted. According to the results, salt stress decreased germination parameters, plumule and radicle lengths, and mitotic index while it increased proline content, lipid peroxidation, and radical contents. Schiff base treatment clearly reduced lipid peroxidation and radical content in all groups. However, it also decreased germination and growth parameters and mitotic index. The obtained results showed that the antioxidant property of this molecule was also preserved in plants under stress, but it was also determined that the molecule had negative effects, primarily on cell division. If necessary modifications can be made to the molecule, the negative effects on cell division can be eliminated, and this molecule, which is very easy and cheap to obtain, may be widely used to increase the tolerance of plants against environmental stress. Сольовий стрес є одним із найважливіших факторів навколишнього середовища, який впливає на сільськогосподарські угіддя та спричиняє втрати продукції. Сьогодні вчені широко досліджують екзогенне застосування різних природних або синтетичних молекул щодо рослин і намагаються підвищити стійкість рослин до стресів навколишнього середовища. У цьому дослідженні молекула основи Шиффа (0, 3, 6 та 9 µM), яка показала антиоксидантні властивості in vitro, була застосована до насіння ячменю під сольовим стресом (0, 50, 150 та 250 mM NaCl). Для того, щоб оцінити вплив цієї молекули на ячмінь в умовах сольового стресу, було досліджено схожість насіння, параметри росту, перекисне окислення ліпідів, вміст проліну, а також гістохімічне виявлення радикалів супероксиду та пероксиду водню та аналіз мітотичного індексу. Згідно з результатами, сольовий стрес зменшував параметри проростання, довжину брунечок та корінців, а також мітотичний індекс, водночас збільшував вміст проліну, перекисне окислення ліпідів та вміст радикалів. Обробка основою Шиффа явно зменшила перекисне окислення ліпідів і вміст радикалів у всіх дослідних групах. Однак водночас це знизило параметри проростання і росту, а також мітотичний індекс. Отримані результати показали, що антиоксидантна властивість цієї молекули збереглася в рослинах під час стресу, але також було визначено, що молекула негативно впливає, насамперед на поділ клітин. Якщо в молекулу було б можливо внести необхідні модифікації, можна було б усунути її негативний вплив на поділ клітин, і тоді цю молекулу, яку дуже легко і дешево отримати, можна було би широко використовувати для підвищення стійкості рослин до стресу навколишнього середовища. M.M. Gryshko National Botanical Garden of the NAS of Ukraine 2022-10-01 Article Article application/pdf https://www.plantintroduction.org/index.php/pi/article/view/1618 10.46341/PI2022018 Plant Introduction; No 95/96 (2022); 85-95 Інтродукція Рослин; № 95/96 (2022); 85-95 2663-290X 1605-6574 10.46341/PI95-96 en https://www.plantintroduction.org/index.php/pi/article/view/1618/1536 Copyright (c) 2022 Sertan Çevik http://creativecommons.org/licenses/by/4.0 |
| spellingShingle | Çevik, Sertan Дослідження потенціалу використання молекули основи Шиффа як екзогенного антиоксиданту для насінні ячменю в умовах сольового стресу |
| title | Дослідження потенціалу використання молекули основи Шиффа як екзогенного антиоксиданту для насінні ячменю в умовах сольового стресу |
| title_alt | Investigating the using potential of Schiff base molecule as an exogenous antioxidant on barley seeds under salt stress conditions |
| title_full | Дослідження потенціалу використання молекули основи Шиффа як екзогенного антиоксиданту для насінні ячменю в умовах сольового стресу |
| title_fullStr | Дослідження потенціалу використання молекули основи Шиффа як екзогенного антиоксиданту для насінні ячменю в умовах сольового стресу |
| title_full_unstemmed | Дослідження потенціалу використання молекули основи Шиффа як екзогенного антиоксиданту для насінні ячменю в умовах сольового стресу |
| title_short | Дослідження потенціалу використання молекули основи Шиффа як екзогенного антиоксиданту для насінні ячменю в умовах сольового стресу |
| title_sort | дослідження потенціалу використання молекули основи шиффа як екзогенного антиоксиданту для насінні ячменю в умовах сольового стресу |
| url | https://www.plantintroduction.org/index.php/pi/article/view/1618 |
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