Salicylic acid with respect to plant growth and

stress

A dissertation submitted for the partial fulfillment of the requirement of the Degree.

MASTER OF SCIENCE

IN

BOTANY

Under the guidance of

Dr. Smrati Mishra

(Assistant Professor)

Department of Botany

MAHARAJA BIJLI PASI GOVT. P.G. COLLEGE

Submitted By: Jaya Pandey

[Link]. Botany Sem. IV

ROLL NO:2110496800009

Department of Botany MAHARAJA BIJLI PASI GOVERNMENT POST GRADUATE COLLEGE, ASHIYANA,
LUCKNOW, Affiliated to, UNIVERSITY OF LUCKNOW, LUCKNOW, UTTAR PRADESH, 226012

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 CERTIFICATE

This is to certify that Jaya Pandey, [Link]. Botany semester IV, Roll No. 2110496800009,
department of Botany, Maharaja Bijli Pasi Govt. P.G. College, Lucknow 226012, India,
has completed her dissertation work on the topic-“Salicylic acid with respect to plant
growth and stress.”

This is also to certify that, the dissertation work has not been submitted for any other degree
and submitting only to the Department of Botany University of Lucknow, India for the
fulfillment of the [Link]. Botany degree.

Candidate’s Signature Mentor’s Signature

JAYA PANDEY Dr. Smrati Mishra

(Assistant prof.)

Department of Botany

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 ACKNOWLEDAGEMENT

I am thankful to God for giving me the strength, knowledge, ability and opportunity to
undertake this study and complete it satisfactorily. I would like to thank and express my
sincere gratitude to my respected advisor, teacher and supervisor, Dr. Smrati Mishra,
Assistant Professor, Department of Botany, Maharaja Bijli Pasi Government P.G. College,
Ashiyana, for her patience, motivation, enthusiasm, immense knowledge, worthy guidance
and professional attitude which is appreciable in completing this dissertation.

I thankfully acknowledge the support and inspiration that I received from my teacher Dr.
Vinay Kumar Prajapati and Dr. Santosh Kumar, for their support, encouragement and
insightful comments.

I am deeply grateful to my parents for their support, encouragement, appreciation and

keen interest in my academic achievements. Without you none of this would indeed be
possible. Finally, I thank my brother for his moral support throughout my dissertation.

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 Table of Contents

1. Introduction

2. Effect of Salicylic Acid on Plant Growth

3.1 SA-Initiated Regulation of Cell Division and Expansion

3.2 SA-Induced Regulation of Plant Growth via Signalling Pathways

3.3 Implications of SA Crosstalk with Other Hormones in Plant Growth Regulation

3. Salicylic Acid-Regulated Physiological Functions

4.1 Seed Germination

4.2 Photosynthesis

4.3 Respiration

4. Role of Salicylic Acid in Biotic and Abiotic Stress Tolerance in Plants

5.1 Role of SA in Biotic Stress

5.1.a Role of SA against Pathogens

5.1.b Role of SA against Herbivores

5.2 Role of SA in Abiotic Stress

5.2.a Salt Stress

5.2.b Drought Stress

5.2.c Heat Stress

5.2.d Cold Stress

5.2.e Heavy Metal Stress

5. Salicylic Acid Biosynthesis in Plants

6. Future Perspectives

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INTRODUCTION

Plants possess intricate mechanisms to regulate their growth and adapt to various
environmental challenges. Hormones play a crucial role in orchestrating these regulatory
processes, governing plant development and mediating responses to both internal and
external cues. Among the diverse array of plant hormones, salicylic acid (SA) has emerged as
a key signalling molecule involved in multiple aspects of plant biology. Salicylic acid (SA) is
an important phytohormone that serves as a critical signal molecule mediating immunity and
plant growth (Hayat et al., 2007; Vlot et al., 2009; Rivas-San Vicente and Plasencia, 2011).
SA is synthesized from chorismate through two distinct pathways in plants, the isochorismate
(IC) and the phenylalanine ammonia-lyase (PAL) pathways (Serino et al., 1995; Zhang and
Li, 2019). Salicylic acid was first identified as an endogenous signal in the resistance
response in tobacco plants in 1979. It was observed that acetylsalicylic acid (aspirin) induced
resistance to tobacco mosaic virus (TMV) by increasing PR protein accumulation and
reducing lesion numbers. Subsequent studies showed that SA levels increase in both infected
and non-infected systemic tissues.

Initially recognized for its role in plant defense against pathogens, particularly in the
activation of systemic acquired resistance (SAR), salicylic acid has since been found to have
far-reaching effects beyond immunity. The understanding of SA has expanded to encompass
its involvement in fundamental processes of plant growth and development, extending its
influence well beyond the realms of plant defense.

The impact of salicylic acid on plant growth is evident through its ability to initiate and
regulate cell division and expansion. SA modulates the delicate balance between cell division
and differentiation, thus influencing overall plant growth and organ development. It acts as a
critical regulator of the cell cycle, influencing the expression of cell cycle regulators and
controlling the progression of cell division. Moreover, SA-mediated signalling pathways
interact with other hormones, including auxins, cytokinins, and gibberellins, integrating
growth signals and fine-tuning growth-related processes such as seed germination,
photosynthesis, and respiration. SA is perceived by the nonexpressor of pathogenesis-related
gene 1 (NPR1) and its paralogues NPR3 and NPR4. It subsequently stimulates the
downstream SA responsive genes and induces plant immune response (Fu et al., 2012; Wu et
al., 2012; Ding et al., 2018). In addition to the canonical SA receptor NPRs

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(NPR1/NPR3/NPR4), there are many other SA-binding proteins (SABPs), such as SABP1
(Catalase), SABP2 (MeSA Esterase), and SABP3 (β carbonic anhydrase), which may act as
potential SA receptors in plants and may be involved in SA signaling (Pokotylo et al.,
2019). SA could be modified through glycosylation, methylation, and amino acid (AA)
conjugation to render it inactive or fine-tune its accumulation, function, and/or mobility
(Dempsey andklessing,2017;Ding&Ding,2020).

Thus affecting the regulation of SA on plant immunity and growth. The final stature of plant
growth is manifested by cell number and cell size, which are mainly controlled by the cell
division and expansion processes, and these are determined by both genetic constraints and
environmental signals (Mizukami, 2001;Weiss et al., 2005; Tsukaya, 2019). In the past two
decades, increasing evidence has shown that SA plays essential roles in regulating plant
growth by affecting cell division and cell expansion (Vanacker et al., 2001; Scott et al.,
2004; Miura et al., 2010; Fujikura et al., 2020). However, the action of SA in the regulation
of plant growth has not been comprehensively described yet in most reviews on this topic,
compared to the role of other plant hormones, such as auxin and gibberellin (GA; Santner et
al., 2009; Wolters and Jürgens, 2009; Depuydt and Hardtke, 2011; Vanstraelen and Benková,
2012; Ali et al., 2020).

Apart from its involvement in the induction in plant growth and development SA has been
shown to improve plant tolerance to major abiotic stresses such as metal (Zhang et al., 2015),
salinity (Khan et al., 2014; Nazar et al., 2015), osmotic (Alavi et al., 2014), drought (Fayez
and Bazaid, 2014), and heat stress (Khan et al., 2013b). Exogenously sourced SA to stressed
plants, either through seed soaking, adding to the nutrient solution, irrigating, or spraying was
reported to induce major abiotic stress tolerance-mechanisms (Horváth et al., 2007; Khan et
al., 2012a,b,c, 2013b, 2014; Anwar et al., 2013; Palma et al., 2013) . This mainly describes
the versatile action and molecular mechanisms of SA regulating plant growth and stress.

Under abiotic stress conditions, such as salt, drought, heat, cold, and heavy metal stresses,
salicylic acid assumes a critical role in conferring stress tolerance to plants. It serves as a
signalling molecule that orchestrates a complex network of physiological and biochemical
responses to counteract the adverse effects of stressors. SA helps plants adapt to challenging
environments by regulating stomatal closure, maintaining cellular homeostasis, activating

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stress-responsive genes, and modulating antioxidant defence systems. These mechanisms
collectively enhance plant resilience and survival under adverse conditions.

Understanding the biosynthesis of salicylic acid in plants is essential for unravelling its
regulatory mechanisms and exploring its potential applications. SA biosynthesis involves a
series of enzymatic steps and can be influenced by various factors, including stress
conditions, hormone interactions, and genetic regulation. Elucidating the intricacies of SA
biosynthesis provides insights into manipulating SA levels in plants, which could have
implications for improving plant growth and stress tolerance.

salicylic acid is a multifaceted hormone that influences plant growth regulation and stress
tolerance. Its involvement in cell division, signalling pathways, physiological functions, and
stress responses highlights its importance in plant biology. The comprehensive understanding
of SA's role presented in this thesis contributes to our knowledge of plant hormone biology
and opens up avenues for harnessing its potential in agriculture to enhance plant growth,
productivity, and stress resilience.

The thesis draws upon published studies and research articles, including those by Bartels and
Sunkar (2005), Mantri et al. (2012), Mohamed et al. (2016), Akladious and Mohamed (2018),
Mohamed et al. (2018), and El-Beltagi et al. (2020), Kaya et al.,( 2023), : Li, A., Sun, X., &
Liu, L. (2022, April 27).These studies have contributed to the understanding of the
multigenic nature of stress tolerance and the utilization of molecular techniques in studying
plant responses to abiotic stresses.

Effect of SA on Plant Growth

Plants may exhibit abnormal growth characteristics as a result of changes in endogenous SA

levels (Rivas-San Vicente and Plasencia, 2011; Pokotylo et al., 2021). Stunted height results
from a high endogenous SA concentration. Many Arabidopsis mutants with
overaccumulating SA, present the dwarf plant phenotype with a shorter stem, smaller
leaves, and/or floral organs compared to the wild type (WT; Bowling et al., 1997; Rate et al.,
1999; Miura et al., 2010). In contrast, the SA-depleted enzyme salicylate hydroxylase (NahG)

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transgenic line and the SA-deficient mutant sid2 in Arabidopsis exhibit more leaf biomass as
compared to WT (Scott et al., 2004; Abreu and Munné-Bosch, 2009).

It's crucial to remember that different plant species or organs may be affected differently by
SA. For instance, the aim1 rice mutant, which has lower endogenous SA levels than the wild
type, has adventitious roots and seedlings that are shorter (Xu et al., 2017). Exogenous SA
has an effect on plant growth depending on the type of the plant and its concentration. In
many plant species, different SA concentrations have either stimulating or inhibitory effects
on plant and organ growth. African violet rosette diameter, leaf quantity, and flower buds
were all increased by 0.01 mm SA treatment, according to research (Jabbarzadeh et al.,
2009), however wheat seedling growth and the production of larger ears were induced by
0.05 mm SA (Shakirova et al., 2003). According to Appu and Muthukrishnan (2014), finger
millet's 0.1mm SA improved blooming and grain set. Kazemi (2013) found that strawberries
with a 0.25mm SA had significantly larger leaves and heavier primary fruits. Furthermore,
0.5mm SA increased the dry weight of the root, shoot, and nodule as well as the number of
flowers and pods in chickpeas, according to Kaur et al. (2022). Additionally, wheat and
mungbean growth and photosynthesis were also significantly [Link],
exogenous SA treatments (0.1 and 1mm) similarly reduced trichome density and number in
Arabidopsis (Traw and Bergelson, 2003), and exogenous SA treatments (0.02-0.03mm)
reduced pollen tube length by around 25% (Rong et al., 2016). While In tobacco, 0.1mm SA
decreased the size of leaf epidermal cells and shoot growth (Dat et al., 2000). Pancheya et
al. (1996) noted that SA concentrations between 0.1 and 1mm hindered the growth of
leaves and roots of barley seedlings in a dosage-dependent manner. In general, for a plant
species, smaller doses of exogenous SA appear to have a growth-promoting effect, whereas
greater amounts may negatively regulate development.

For instance, 0.25mm SA considerably reduced the growth of chamomile roots and rosette
leaves by 40 and 43%, respectively, while 0.05mm SA dramatically increased both of these
growth rates by 32 and 65% (Kovácik et al., 2009). According to Pasternak et al. (2019), all
root growth processes were reduced in Arabidopsis when low-concentration SA (0.05mm)
was applied. Inconsistent growth responses to different SA concentrations were also seen in
wheat and pepper (Hayat et al., 2005; Canakci, 2011). It should be noted that the threshold
SA concentrations between growth promotion and growth inhibition may vary among plant

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species. These inconsistent results caused by different exogenous SA concentrations imply
that this compound has a variety of functions in plant growth.

Fig:1 Descriptive model of salicylic acid function in plant growth and development. SA is perceived
by NPR1, a transcriptional activator that regulates gene expression that might participate in seed
germination, flowering, and/or senescence regulation. In addition, SA is a key regulator of plant cell
redox status by inhibiting catalase and peroxidase activity, and thus modulating reactive oxygen
species (ROS) levels. The positive effect of SA on photosynthesis contributes to electron acceptor
availability and redox status. NPR1 oligomerization is redox modulated (Rivas-San Vicente &
Plasencia, 2011)

SA-INITIATED CELL DIVISION AND EXPANSION REGULATION

By altering cell division and expansion, either in a negative or positive way, SA can control
how quickly a plant grows. Some evidence suggests that SA has a detrimental impact on the
two cellular processes in Arabidopsis leaves. For instance, SA-deficient NahG transgenic
plants grew faster than WT plants and had 1.7 times as much leaf rosette biomass in the early
stages of reproduction. (Abreu and MunnéBosch, 2009). This increased growth effect on
NahG transgenic plants was more obvious at low temperatures, and it resulted from enhanced
cell expansion of rosette leaves. (Scott et al., 2004; Xia et al., 2009). Additional research has
shown that NahG transgenic plants at 4°C displayed an enhanced expression of the cell cycle
G1/S transition compared to WT. bigger cells were produced as a result of the cyclin D 3

9
(CYCD3) regulator and increased endoreduplication levels. (Xia et al., 2009). This evidence
indicated that SA suppresses cell expansion by regulating the expression of the cyclin genes.
In addition, the SA-accumulating mutant siz1 showed a dwarf phenotype characterized by a
decreased leaf cell volume and number. The cell division and expansion defects caused by
siz1 can be suppressed through the overexpression of NahG (Miura et al., 2010), further
demonstrating that SA inhibits these two cellular processes. However, SA accumulation in a
different context may exhibit a discrepant action on cell growth control. Vanacker et al.
(2001) found that SA activates cell division and expansion in acd6-1 leaves with a very high
SA level. Additionally, the positive effect of SA on cell division was also found in the roots.
For example, the aim1 rice mutant, whose SA biosynthesis was deficient, showed a reduced
root meristem activity and a significantly lower expression of the cell cycle G2/M phase
transition regulator cyclin B1;1 (CYCB1;1) compared to WT, indicating that SA has a
positive role in root cell division (Xu et al., 2017). Moreover, the SA-overaccumulating
Arabidopsis mutant known as constitutively activated cell death 1 (cad1) increased cell
division in the quiescent centre (QC) in the root apical meristem, which was observed in the
SA-treated WT (Wang et al., 2021). In summary, the SA-induced regulation of cell division
and expansion is complicated and may depend on plant organs and the context in which
signalling occurs.

SA-INDUCED REGULATION OF PLANT GROWTH THROUGH A VARIATION

OF SIGNALLING PATHWAYS

It has been extensively studied how SA signalling occurs during plant immune responses, and
NPR1 has been identified as a crucial step in this procedure. NPR1 is crucial for SA-mediated
growth, too. regulation via regulating cell division and growth (Vanacker et al., 2001;
Fujikura et al., 2020; Wang et al., 2021). Less cells and more DNA were found in the leaves
of the Arabidopsis npr1-1 mutant, demonstrating that NPR1 encourages cell proliferation and
inhibits endoreduplication in leaves. es (Vanacker et al., 2001). Fujikura et al. (2020) found
that the xs2 mutant accumulated high SA contents and impaired cell expansion, producing
smaller cells compared to those observed in WT. It's interesting to note that the xs2 npr1

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double mutant reversed the considerable cell size deficiency seen in the xs2 mutant. These
findings suggest that the inhibition of cell growth was carried out by an NPR1-dependent
signalling pathway in xs2. Additionally, the SA-overaccumulating Arabidopsis Additionally,
the SA-overaccumulating Arabidopsis mutant cad1 increased cell division in the QC, which
was rescued through the mutation of SID2 or NPR1, indicating that the QC cell division in
the cad1 mutant is promoted in an NPR1-dependent SA signalling pathway (Wang et al.,
2021). Further research revealed that SA build up in the cad1 mutant facilitates QC cell
proliferation by increasing reactive oxygen species and downregulating transcription.
Plethora 1 (PLT1), PLT2, and WUSCHEL-related homeobox 5 (WOX5) factor genes are
essential for QC upkeep (Wang et al., 2021).

NPR1 may also have a negative effect on cell division in some specific developmental
contexts, such as the SA-Ethylene (ET)-mediated apical hook development (Raz and
Koornneef, 2001; Huang et al., 2020). The above examples support the hypothesis that SA
could regulate cell division, cell expansion, and then plant growth in an NPR1-dependent
manner. Besides the NPR family proteins, other SABPs also play essential roles in SA-
mediated plant growth regulation (Pokotylo et al., 2019). Tan et al. (2020) found that SA
directly binds to A subunits of protein phosphatase 2A (PP2A) to suppress the
dephosphorylation of PIN-formed (PIN) auxin efflux carriers and inhibit root development
(including root elongation, gravity response, and lateral root formation) in an NPR-
independent manner. Additionally, the inhibiting effect of SA on pollen tube growth in
Arabidopsis is also independent of NPRs because the npr1, npr3, npr4, and npr3 npr4 mutants
exhibited responses that were identical to that of 20mm SA in WT (Rong et al., 2016).
Further investigations revealed that SA and methylated SA (MeSA) antagonistically regulate
pollen tube growth by affecting clathrin-mediated endocytosis (CME) and the apical
activation of a key pollen tip growth regulator, known as Rho-type GTPase 1 (ROP1), with
SA and MeSA having inhibitory and stimulatory effects, respectively. Interestingly, the
methylesterase and methyltransferase enzymes, which catalyze the interconversion between
SA and MeSA, are both localized on the tip of pollen tubes, indicating that the tip-localized
production of the two compounds plays an important role in the regulation of polar cell
growth (Rong et al., 2016). However, it remains to be determined which SABP is
responsible for the SA/MeSA-mediated regulation of pollen tube growth.

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There have also been reports of additional SA mechanisms that control plant development. It
was discovered that the Arabidopsis cpr5 with an increased SA level had shorter primary
roots and lateral roots .The cpr5/sid2 double mutant was able to restore the root number
(Bowling et al., 1997). Further investigation revealed that CPR5 can suppress the buildup of
SA and promote development by inhibiting According to Meng et al. (2017), the SA-
dependent IRE1-bZIP60 signalling pathway controls plant unfolded protein response (UPR),
which inhibits organ growth. This finding suggests that CPR5 controls root growth through
this mechanism. By altering the expression of genes involved in cell proliferation, SA also
controls plant growth. The above-mentioned siz1 mutant's SA accumulation negatively
controls the ability of the genes xyloglucan endotransglycosylase/hydrolase (XTH), XTH8
and XTH31, to be expressed, which promotes leaf cell growth and loosening of the cell wall
(Miura et al., 2010). Nevertheless, the SABPs involved in these pathways have not been
explored yet.

Thus the effect of SA on organ and plant growth is mediated by several receptors and
happens through a variety of routes. That SA has a complicated role in controlling plant
development.

PLANT GROWTH REGULATION IMPLICATIONS OF SA CROSSTALK WITH

OTHER HORMONES

In order to influence the growth of plants and organs, SA also interacts with other hormones
that control cell division and expansion, such as auxin, GA, and ethylene (ET). h (Ari et al.,
2020; Emamverdian et al., 2020; Mazzoni-Putman et al., 2021; Pokotylo et al., 2021).

Auxin is a major growth hormone that controls these cellular processes, especially in roots
(Perrot-Rechenmann, 2010; Barbeza et al., 2017; Huang et al., 2019; Seo et al., 2021), and
SA can influence root development by affecting the accumulation and transport of auxin.

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Pasternak et al. (2019) proved that exogenous SA affects the root tip meristem in
Arabidopsis in a concentration-dependent manner. A low concentration of SA (below 0.05
mM) causes auxin accumulation via promoting the production of the enzyme TRP, which
produces auxin via inhibiting PIN2 and PIN7, the auxin efflux protein PIN1, and the
Arabidopsis 1 aminotransferase (TAA1). Then, through a cyclin D6;1 (CYCD6;1)-dependent
mechanism, it boosts the number of periclinal and tangential cells in the outer layers of the
roots. A high dose of SA, however (over 0.05 mM), on the other hand, causes auxin depletion
in the root meristem by suppressing the expression of PIN1, PIN2, and PIN7 and thus
preventing the cell cycle and growth of the roots (Pasternaket al., 2019).

By directly interacting with PP2A to suppress its dephosphorylation activity towards PIN2,
SA can also cause PIN2 to become hyperphosphorylated. activity, which causes auxin export
to decline and root growth to slow (Tan et al., 2020). Additionally, SA can prevent auxin by
inducing PIN2 hyperclustering via a lipid nanodomain organisation dependent on remorin
(REM). accrual and deterioration of the auxin-mediated root gravitropic response (Ke et al.,
2021).

GA promotes plant growth by increasing cell division and expansion (Achard et al., 2009;
Sprangers et al., 2020; Vercruysse et al., 2020). The application of exogenous GA increases
SA biosynthesis in wild Arabidopsis plants or promotes seed germination and seedling
growth in the sid2 under adverse conditions, suggesting a synergistic interplay between SA
and GA (Alonso-Ramírez et al., 2009). However, SA and GA can clash when it comes to
controlling growth. Treatments with SA in barley were found to reduce the expression of
alpha-amylase that GA causes. potentially affect seed germination and subsequent growth in
the aleurone layers (Xie et al., 2007).

In Arabidopsis, GA increases trichome number, while this effect is suppressed by SA (Traw

and Bergelson, 2003). Furthermore, the Arabidopsis hydroxycinnamoyl CoA: shikimate
hydroxycinnamoyl transferase (HCT)-RNAi lines are characterised by The plant suffered
from severe dwarfism due to excessive SA accumulation, which decreased the expression of
GA marker genes. By altering SID2 or overexpressing NahG, the abnormalities in GA
signalling and growth in HCT RNAi lines may be corrected. (Gallego-Giraldo et al., 2011). It
is therefore inferred that SA negatively regulates cell division, cell expansion, and overall
plant growth, by antagonizing GA. ET is also an endogenous regulator of cell division and

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expansion in vegetative growth (Dubois et al., 2018). It participates to the stimulation of cell
division during the early development of apical hooks (Raz and Koornneef, 2001). SA can
inhibit ET biogenesis and signalling (Leslie and Romani, 1988; Huang et al., 2020) and, in
Arabidopsis, it was shown to reduce the apical hook angle of etiolated seedlings by
antagonizing ET signalling.

The key transcription factor of ET, ethylene insensitive 3 (EIN3), directly interacts with the
SA-activated NPR1, disrupting its binding to its target gene promoters. such as the promoter
of HOOKLESS 1, which is crucial for developing the apical hook and hence lowering the
hook angle (Huang et al., 2020). Consequently, by opposing ET, SA may have a detrimental
influence on cell division.

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 Fig2:(Li A, Sun X and Liu L (2022) Action of Salicylic Acid on Plant Growth. Front. Plant
Sci. 13:878076. doi: 10.3389/fpls.2022.878076)

SA-regulated physiological functions

Seed germination

Exogenous SA's impact on seed germination. Seed germination is influenced by

environmental conditions as well as interactions among the plant hormones abscisic acid
(ABA), jasmonic acid (JA), gibberellins (GA), ethylene (ET), brassinosteroids (BR), auxins
(AUXs), and cytokinins (CKs). Conflicting data imply that SA can either hinder germination

15
or boost seed vigour, raising questions about its function in the germination of seeds. The SA
concentrations used can explain the apparent contradictory effects.

SA concentrations greater than 1 mM can suppress or delay germination in A. thaliana

(Rajjou et al., 2006). Barley seed germination is inhibited by SA dosages more than 0.250
mM (Xie et al., 2007), whereas maize seed germination is entirely prevented by SA doses
between 3 and 5 mM (Guan and Scandalios, 1995). The unfavourable impact of SA on seed
germination is most likely caused by an oxidative stress that SA causes. Hydrogen peroxide
(H2O2) levels in SA-treated Arabidopsis plants rise by up to three times as a result of
increased Cu, Zn-superoxide dismutase activity and inactivation of the H2O2-degrading
enzymes catalase and ascorbate peroxidase.

SA enhancement of abiotic stress-induced seed germination. It's interesting to note that SA

considerably enhances Arabidopsis seed germination and seedling establishment when
administered exogenously in low dosages under various abiotic stress conditions (Rajjou et
al., 2006; Alonso-Ramrez et al., 2009).

Only 50% of Arabidopsis seeds germinate under salt stress (100–150 mM NaCl), but in the
presence of SA (0.05–0.5 mM), seed germination rises to 80%. The inhibitory effects of heat
stress (50 C for 3 hours) and oxidative stress (0.5 mM paraquat) on seed germination are also
partially reversed by exogenous administration of SA (Alonso-Ram'rez et al., 2009).

These findings are consistent with the Arabidopsis sid2 mutant's delayed germination
phenotype under high salinity (Alonso-Ram'rez et al., 2009).

Low SA levels are present in this mutant because the isochorismate synthase gene has been
compromised. However, germination is unaffected by high salinity, and NahG transgenic
lines expressing a bacterial salicylate hydroxylase similarly have lower SA levels than wild-
type plants (Borsani et al., 2001). This apparent disparity is caused by catechol's antioxidant
properties, which accumulate in NahG seeds and seedlings as a result of salicylate
hydroxylase (Lee et al., 2010). In high salinity environments, SA thus promotes
germination by minimising oxidative damage. Additionally, proteomic investigations
revealed that SA induces two superoxide dismutases in Arabidopsis germination seeds, which
may help to increase antioxidant capacity (Rajjou et al., 2006). A strong up-regulation of
translation initiation and elongation factors, proteases, and two subunits of the 20S
proteasome is also induced by SA treatment (0.5 mM for 24 h), supporting the idea that SA

16
enhances seed germination by encouraging the synthesis of proteins required for germination
and the mobilisation or degradation of seed proteins accumulated during seed maturation.
Additionally, SA strongly stimulates the biosynthesis of a number of enzymes involved in
metabolic pathways including the glyoxylate cycle, pentose phosphate pathway, glycolysis,
and gluconeogenesis, indicating that SA aids in the seedling's transition from a dormant state
to the establishment of a robust seedling (Rajjou et al., 2006).

ABA and GAs interact with SA during germination. The fate of germination at this early
developmental stage is determined by a complicated interplay between SA and both ABA
and GAs. GAs play a part in SA biosynthesis and the SA pathway in Arabidopsis. In
comparison to seeds ingested in water and wild-type plants, the ingestion of 50 lM GA3 by
seeds for 24 hours and the overexpression of a GA-stimulated gene from beechnut
(FcGASA4) in Arabidopsis plants cause a 2-fold increase in SA levels.

Additionally, FcGASA4-overexpressing lines and Col0 seedlings cultivated in the presence

of GA3 exhibit enhanced expression of the ICS1 (isochorismate synthase) and NPR1
(nonexpressor of PR-1) genes, implicated in SA biosynthesis and perception, respectively.

Intriguingly, under 150 mM NaCl stress, exogenous SA (50 lM) only minimally improves the
germination of the sid2 mutants but exogenous GA3 (50 lM) significantly recovers seed
germination in the GA-deficient mutant ga1-3 (Alonso-Ram'rez et al., 2009). Despite the fact
that these findings point to a synergistic interaction between SA and GA, it was shown that
SA and GA had an antagonistic relationship when it came to barley germination, which may
be explained by using a greater dose of SA. The inhibition of barley seed germination and
post-germination growth by SA is accompanied by suppression of GAinduced a-amylase
(Amy32b) expression through induction of a WRKY repressor (HvWRKY38). Expression of
HvWRKY38 in aleurone cells is down-regulated by GAs, but up-regulated by SA and ABA,
so this transcription factor might serve as a converging node of the SA and ABA signal
pathways involved in suppressing GA-induced seedgermination (Xie et al., 2007). Additional
evidence supporting the cross-talk between ABA and SA signalling is the increased synthesis
of ABA-regulated proteins, such as late embryogenesis abundant (LEA) proteins, dehydrins,
and heat shock proteins, in Arabidopsis seeds germinated in the presence of 0.5 mM SA
(Rajjou et al, 2006). The ubiquitin–proteasome system (UPS) pathway in hormone signalling
integration. Because GAs and ABA have opposing roles in the regulation of germination, SA
might act as a rheostat contributing with both hormones. Recent biochemical evidence points

17
to the UPS as a mechanism to balance the antagonic control of seed germination between
ABA and GAs (Zentella et al., 2007; Piskurewics et al., 2008). In fact, in the UPS, many
hormone signalling pathways converge (Santner and Estelle, 2009; Vierstra, 2009; Santner
and Estelle, 2010) thus influencing many aspects of plant growth and development. Recently
it was found that NPR1, the key transducer of SA signalling in plant defence responses,
associates in the nucleus with Cullin3-based E3 ligases and other components of the COP9
signalosome, which controls proteasomal degradation. Moreover, NPR1 proteasome-
mediated turnover is promoted by SA-induced phosphorylation of the Ser11 and Ser15
residues (Spoel et al., 2009). Initially, it was found that the activity of NPR1 is regulated by
its subcellular localization, because the transcriptional co-activator is predominantly
sequestered in the cytoplasm as an oligomer, but in pathogen-infected cells SA accumulation
promotes partial reduction of the NPR1 oligomer to a monomer, which is targeted to the
nucleus by a bipartite nuclear localization sequence (Mou et al., 2003). Interestingly, NPR1
also enters the nucleus when basal SA levels are low and no infection is occurring, and it has
been suggested that it may regulate additional genes. If this scenario could be confirmed, it
would be interesting to analyse the contribution of these genes to the regulation of
germination, plant growth, and development.

It is also worth mentioning the role of the DELLA proteins as potential integrators of
phytohormone signalling in the regulation of germination, cell redox state, growth, and stress
responses (reviewed by Smirnoff and Grant, 2008; Grant and Jones, 2009; Harberd et al.,
2009). DELLA proteins are repressors of GA signalling and, in turn, GA derepresses its
pathway by promoting proteasomal degradation of the DELLA protein RGA (Dill et al.,
2004). There are five DELLA genes in Arabidopsis: GAI (GA insensitive), RGA (repressor
of GA1-3), RGL1, RGL2, and RGL3 (RGA-like). RGL2 is considered to be the main
DELLA factor repressing germination (Lee et al., 2002; Tyler et al., 2004), although the other
DELLA genes also contribute to regulate germination (Cao et al., 2005). Recently, it was
found that RGL2 expression is strongly stimulated by ABA, and that RGL2 protein is
necessary to elevate endogenous ABA and ABI5 (another germination repressor) expression
levels, specifically when GA levels are low (in ga1-3 mutants, or in the presence of GA
synthesis inhibitors). Moreover, RGL2 is necessary to repress testarupture (Piskurewicz et al.,
2008). Other phytohormones such as AUXs and ET also modulate plant growth and
morphogenesis through a DELLA-dependent mechanism (Achard et al., 2003, 2006, 2007;
Fu and Harberd, 2003). Interestingly, DELLAs modulate the balance of SA/JA signalling in

18
disease resistance, promoting JA perception and/or signalling, and repressing SA biosynthesis
and signalling (Navarro et al., 2008). However, it is necessary to determine whether this
modulation of SA/JA signalling by DELLA proteins also occurs during growth and
development. DELLAs also modulate the levels of reactive oxygen species (ROS), which are
also involved in growth-regulatory mechanisms (Achard et al., 2008). Because ROS are
closely associated with SA signalling in an autoamplification loop (Shirasu et al., 1997), it
has been proposed that the attenuation of SA signalling by DELLAs is the result of
diminishing ROS levels (Grant and Jones, 2009).

Photosynthesis

Photosynthesis is a vital process in plants that is influenced by various regulatory factors.

Salicylic acid (SA) has emerged as an important regulator of photosynthesis, affecting leaf
and chloroplast structure, stomatal closure, chlorophyll and carotenoid contents, as well as
the activity of key enzymes such as RuBisCO (ribulose-1,5-bisphosphate
carboxylase/oxygenase) and carbonic anhydrase. This review explores recent evidence
highlighting the diverse effects of SA on photosynthesis and its underlying mechanisms. The
findings reveal dose-dependent responses of plant species to exogenous SA, with high
concentrations leading to reduced photosynthetic rate and RuBisCO activity. Conversely,
lower concentrations of SA have been shown to enhance photosynthetic CO2 assimilation.
Furthermore, SA plays a crucial role in protecting plants against oxidative stress induced by
herbicides and heavy metals. SA also contributes to light acclimation and redox homeostasis,
facilitating optimal photosynthetic activity in response to changes in light conditions.
Additionally, SA has been implicated in stomatal closure, acting in conjunction with other
phytohormones to regulate this process. This comprehensive review sheds light on the
multifaceted role of SA in modulating various aspects of photosynthesis and highlights its
significance in plant physiology.

Photosynthesis is a fundamental process in plants, enabling them to convert sunlight into

chemical energy. The regulation of photosynthesis involves complex interactions between
various signalling molecules and environmental factors. Salicylic acid (SA), a plant hormone
known for its role in defense against pathogens, has recently emerged as a key regulator of

19
photosynthesis. SA has been shown to influence leaf and chloroplast structure, stomatal
closure, pigment contents, and the activity of important enzymes such as RuBisCO.
Understanding the effects of SA on these aspects of photosynthesis is crucial for unravelling
its role in plant physiology.

Effects of SA on Leaf and Chloroplast Structure: Recent studies have demonstrated that
exogenous SA can induce alterations in leaf anatomy, including a reduction in the width of
the epidermis and mesophyll tissue. These structural changes correlate with an increase in
chloroplast volume, swelling of grana thylakoids, and coagulation of the stroma. The
observed ultrastructural changes in chloroplasts suggest that SA affects thylakoid membranes
and the light-induced reactions associated with them. These structural modifications may
contribute to the diminished photosynthetic activity observed at high concentrations of SA.

Impact of SA on RuBisCO Activity: RuBisCO is a key enzyme involved in carbon fixation

during photosynthesis. Studies have shown that high concentrations of SA can lead to a
reduction in RuBisCO activity in barley plants. This decline in activity has been attributed
to decreased protein levels compared to non-treated plants. Furthermore, total soluble protein
content is also reduced. These findings suggest that the negative impact of high SA
concentrations on photosynthetic rate may be associated with impaired RuBisCO activity and
protein synthesis.

Role of SA in Stomatal Closure: Stomatal closure is an essential process that regulates gas
exchange and water loss in plants. SA has been implicated in the regulation of stomatal
closure, particularly during pathogen attack. Experiments with Arabidopsis have shown that
SA induces rapid stomatal closure within a few hours. Moreover, SA-deficient mutants
display compromised stomatal closure upon pathogen perception. The interaction between
SA and abscisic acid (ABA), another phytohormone involved in stomatal regulation, seems to
be crucial for promoting stomatal closure in response to pathogen attack.

SA as a Modulator of Light Acclimation and Redox Homeostasis: Light acclimation is a

crucial process by which plants optimize their photosynthetic efficiency under varying light
conditions. Recent studies have highlighted the role of SA in light acclimation processes.
Arabidopsis plants with high constitutive SA levels exhibit altered photosynthetic parameters
under different light conditions. These plants display decreased maximum efficiency of
photosystem II (PSII), reduced quantum yield of PSII, increased thermal dissipation of
absorbed light energy, and reduced stomatal conductance under low light conditions.

20
Conversely, plants with low SA content exhibit impaired acclimation to high light,
characterized by increased oxidative stress and enhanced thermal energy dissipation. These
findings suggest that SA signalling pathways interact with other hormonal and reactive
oxygen species signalling pathways to regulate light acclimation processes and maintain
redox homeostasis.

SA-Mediated Protection against Oxidative Stress: SA has been shown to confer protection
against oxidative stress induced by herbicides and heavy metals, which can impair
photosynthetic activity. Pre-treatment with SA has been found to reduce chlorophyll losses,
reactive oxygen species (ROS) production, lipid peroxidation, and electrolyte leakage,
thereby preserving photosynthetic activity. SA's ability to mitigate oxidative stress is
attributed to its antioxidant properties and its modulation of the antioxidant defense system.

Thus Salicylic acid plays a multifaceted role in regulating photosynthesis, influencing leaf
and chloroplast structure, RuBisCO activity, stomatal closure, and light acclimation
processes. The effects of SA on photosynthetic parameters are concentration-dependent and
can vary among plant species. Further research is needed to unravel the precise molecular
mechanisms underlying SA's effects on photosynthesis and explore its potential applications
in improving plant productivity and stress tolerance. Understanding the intricate interactions
between SA and other signalling molecules involved in photosynthesis will contribute to a
more comprehensive understanding of plant physiology and potentially lead to the
development of strategies for enhancing crop productivity and stress resilience.

Respiration

The role of salicylic acid (SA) in regulating the alternative oxidase (AOX) pathway in plants
has been investigated. SA has been found to induce the expression of AOX genes in both
thermogenic and non-thermogenic plants, leading to increased cyanide-resistant oxygen
uptake and heat evolution. The concentration-dependent effect of SA on NtAOX1 gene
expression correlates with protein abundance, with a significant increase observed after 4
hours of treatment. However, gene expression returns to basal levels after 24 hours.

The AOX pathway is involved in the oxidation of ubiquinol and the reduction of molecular
oxygen, independent of cytochrome oxidase pathway inhibitors. This non-proton-driven
carrier provides flexibility in ATP synthesis control to maintain growth rate homeostasis and

21
serves as a potential target for SA-mediated plant growth regulation. Moreover, AOX is
believed to limit the production of reactive oxygen species (ROS) in mitochondria.
Overexpression of AOX reduces ROS levels, while suppressing AOX results in a significant
increase in ROS abundance.

In addition to inducing the alternative respiration pathway through AOX gene expression, SA
also affects mitochondrial electron transport and oxidative phosphorylation. Low
concentrations of SA stimulate respiration in whole cells and isolated mitochondria by acting
as an uncoupler. However, higher concentrations inhibit respiration by preventing electron
flow from substrate dehydrogenases to the ubiquinone (UQ) pool. These inhibitory effects
may be attributed to SA's phenolic nature, acting as a quinone analogue.

The impact of SA on mitochondrial function extends beyond tobacco plants, as similar

uncoupling and inhibitory effects have been observed in soybean mitochondria and
mammalian mitochondria. Notably, the inhibition of respiration by SA leads to a decrease in
ATP levels, which is evident shortly after treatment and does not require de novo protein
synthesis.

SA regulates the AOX pathway in plants by inducing AOX gene expression, which results in
increased cyanide-resistant oxygen uptake and heat evolution. Additionally, SA affects
mitochondrial function by acting as an uncoupler at low concentrations and inhibiting
electron flow at higher concentrations. These findings contribute to our understanding of the
intricate regulatory mechanisms involving SA and the AOX pathway, with potential
implications for plant growth and ROS management in various plant species.

Role of Salicylic Acid in Biotic and Abiotic Stress Tolerance in Plants

Abiotic stresses, such as drought and salinity, have a significant impact on plant productivity
and distribution, affecting over 10% of arable land globally. These stresses, coupled with the
increasing trends of desertification and salinization, have led to a decline in average crop
yields by more than 50%. Traditional plant breeding approaches have had limited success due
to the complex multigenic nature of stress tolerance. However, in the past decade, molecular

22
techniques have provided valuable insights into the mechanisms by which plants perceive
environmental signals and activate adaptive responses.

This thesis aims to explore the potential of secondary metabolites in improving plant
performance and reducing productivity losses under abiotic stress conditions. Specifically, it
investigates the role of salicylic acid (SA), a phenolic compound, in activating plant defenses
and enhancing systemic acquired resistance (SAR).

The thesis draws upon published studies and research articles, including those by Bartels and
Sunkar (2005), Mantri et al. (2012), Mohamed et al. (2016), Akladious and Mohamed (2018),
Mohamed et al. (2018), and El-Beltagi et al. (2020), Kaya et al.,( 2023), : Li, A., Sun, X., &
Liu, L. (2022, April 27).These studies have contributed to the understanding of the
multigenic nature of stress tolerance and the utilization of molecular techniques in studying
plant responses to abiotic stresses.

Secondary metabolites, including phenolics, terpenes, and nitrogen/sulfur-containing

compounds, play a vital role in plant growth, development, and defense against biotic and
abiotic stresses. Phenolic compounds, such as SA, have been identified as key regulators of
plant defenses, particularly in activating SAR. SA, and its derivative aspirin (acetyl SA), have
been extensively used as anti-inflammatory drugs and have shown promise in enhancing
plant resilience to abiotic stresses.

The application of secondary metabolites, particularly SA, offers a potential approach to

improve plant performance and mitigate the loss of productivity under abiotic stress
conditions. Understanding the mechanisms by which these compounds interact with plant
systems can inform the development of strategies to enhance stress tolerance in crops.

The complex defense system of plants involves the production of secondary metabolites,
including phenolics, terpenes, and nitrogen/sulfur-containing compounds. Among these, SA
has emerged as a key player in activating plant defenses and systemic acquired resistance.
Harnessing the potential of secondary metabolites, such as SA, through molecular techniques
holds promise for improving crop tolerance to abiotic stresses and increasing agricultural
productivity in the face of environmental challenges.

Role of SA in Biotic Stress

Role of SA against Pathogens

23
The role of salicylic acid (SA) in plant defense mechanisms against pathogens. Early studies
in the 20th century revealed that when plants were infected by pathogens, systemic defense
mechanisms were activated, leading to increased resistance against subsequent pathogen
attacks. One of the early responses associated with this defense is the hypersensitive response
(HR), which involves the formation of necrotic lesions at the site of pathogen entry,
accumulation of reactive oxygen species (ROS), and activation of defense-related genes,
including pathogenesis-related (PR) proteins.

PR proteins are encoded by several families of PR genes, and their increased expression is
observed in non-inoculated tissues during pathogen infection. This leads to the development
of systemic acquired resistance (SAR), which provides broad resistance against different
pathogens. Among the PR genes, PR1, PR2, and PR5 are strongly induced by biotrophic and
semi-biotrophic pathogens. The expression of these genes is dependent on SA, and they are
often used as markers for the SA pathway. PR1 has been found to inhibit pathogen growth by
sequestering sterol from pathogens.

Salicylic acid was first identified as an endogenous signal in the resistance response in
tobacco plants in 1979. It was observed that acetylsalicylic acid (aspirin) induced resistance
to tobacco mosaic virus (TMV) by increasing PR protein accumulation and reducing lesion
numbers. Subsequent studies showed that SA levels increase in both infected and non-
infected systemic tissues. SA also plays a role in "priming," which is the faster and stronger
activation of defense mechanisms in response to pathogen or microbial elicitors.

Salicylic acid affects the activity of mitochondrial alternative oxidase, an enzyme involved in
the reduction of oxygen to water without the synthesis of ATP. This effect of SA on
alternative oxidase has been associated with SA-mediated resistance to certain viruses. SA
also binds to various enzymes involved in reactive oxygen species (ROS) metabolism and
redox homeostasis, such as catalase, ascorbate peroxidase, aconitase, and carbonic anhydrase.
These interactions influence the biochemical activity of these enzymes and may contribute to
plant defense responses.

Salicylic acid is known to induce the expression of defense-related genes, promote H2O2
production, and activate cell death processes. It can also bind to specific regulatory elements
in the promoters of defense-related genes. SA has been shown to increase the as-1-binding
activity and activate promoters containing as-1-like elements. It can activate MAP kinases

24
and regulate the expression of genes encoding antioxidants, thereby maintaining the redox
state of the cell.

Furthermore, SA accumulation leads to the monomerization of NPR1, a transcription factor

involved in the regulation of PR gene expression. Monomeric NPR1 is transported to the
nucleus, where it promotes the binding of transcription factors to SA-responsive promoters,
resulting in the expression of PR genes. NPR1 is considered a positive regulator of SAR and
is required for the translation of the SA signal and expression of resistance genes.

In summary, salicylic acid plays a crucial role in plant defense against pathogens. It activates
systemic defense responses, induces the expression of defense-related genes, regulates redox
homeostasis, and promotes the accumulation of reactive oxygen species. SA-mediated
signalling pathways contribute to the establishment of systemic acquired resistance and the
inhibition of viral replication and transmission in plants.

Role of SA Against Herbivores

Salicylates are also responsible for protecting plants from herbivores by stimulating affected
plants (plants experiencing insect damage) to increase the production of their volatile
secretions which are responsible for attracting anti-herbivores (Thaler 1999). Responses to
SA depend on a regulatory protein called Nonexpressor of Pathogenesis-Related Genes1
(NPR1) (Pieterse and Van Loon 2004). The NPR1 gene is activated through redox pathways
by SA accumulation and is translocated to the nucleus; however, it does not bind to DNA
directly, but acts through transcription factors (Pieterse and Van Loon 2004). SA induces
greater defence against 23 Role of Salicylic Acid in Biotic and Abiotic Stress Tolerance in
Plants 539 piercing and sucking type of insect pests than the chewing ones (Zhao et al. 2009).
SA signalling molecule is involved in local defense as well as induction of systemic
resistance. Production of ROS by SA pathway has been proposed to induce resistance in
plants against insect pests, e.g., in tomato plants against Helicoverpa armigera (Peng et al.
2004). H2O2 induced by SA in plants defends them against various insect pests since H2O2

25
actively damages the digestive system of insects leading to reduced growth and development
(Peng et al. 2004). Furthermore, SA signals the release of plant volatiles that attract the
natural enemies of insect pests, e.g., lima bean and tomato plants infested by spider mite
attract the natural enemies of spider mite (de Boer et al. 2004). However SA and jasmonic
acid (JA) act antagonistically, where SA inhibits the activity of JA and vice versa (Maffei et
al. 2007). SA serves as a volatile signal to trigger induced defenses in plants, including HIPV
emission, and a number of predaceous arthropods are attracted to SA under field conditions
(de Boer et al. 2004; Maffei et al. 2007).

Role of SA in Abiotic Stress

The role of SA in mitigating abiotic stress has widely been studied in the last few decades. A
large volume of research reports indicate that both endogenous SA synthesis and exogenous
application enhance plant tolerance to salinity (Hasanuzzaman et al. 2014), drought (Kang et
al. 2012; Alam et al. 2013), extreme temperature (Kumar et al. 2015; Khanna et al. 2016),
and toxic metal and metalloids (Singh et al. 2015; Islam et al. 2016). Exogenous SA showed
enhanced plant growth, photosynthesis, and decreased ROS production under various abiotic
stresses .

Salt Stress

Exogenous salicylic acid (SA) treatments have been shown to have a significant impact on
plant responses to sodium chloride (NaCl) stress. However, the effectiveness of these
treatments depends on various factors, including SA concentrations, plant species, application
mode, plant physiological state, salinity level, and exposure time to NaCl. This thesis aims to
provide a comprehensive summary of the research conducted on the effects of some of the
endogenous SA effect as mentioned above in the thesis while concentrating on exogenous SA
treatments in improving plant growth and seed germination under different stresses like salt
stress .

Plant Growth Enhancement Several studies have demonstrated that exogenous SA treatments
improve plant growth in the presence of NaCl stress. Notably, Szepesi (2006), Bastam et al.
(2013), and Liu et al. (2014) reported positive effects of SA treatments on plant growth under

26
saline conditions. For instance, pre-treating tomato plants with low SA concentrations (10^-4
M) enhanced their acclimation to 100 mM NaCl, resulting in improved photosynthetic
efficiency and increased activity of ascorbate peroxidase and guaiacol peroxidase in roots.
Furthermore, SA pre-treatment induced the accumulation of polyamines, which contributed
to plant growth promotion under salinity stress.

Seed Germination Process Exogenous SA treatments have also been shown to enhance the
seed germination process in the presence of NaCl. Rajjou et al. (2006) and Lee et al. (2010)
observed that SA treatment improved seed germination under saline conditions. These
findings suggest that SA plays a crucial role in alleviating the negative effects of salt stress on
seed germination, thereby enhancing the establishment of plants in saline environments.

Mechanisms of Action The protective effects of SA against salinity stress in plants are
attributed to various physiological and biochemical mechanisms. Tari et al. (2002, 2004) and
Szepesi et al. (2005) reported that SA pre-treatment increased the activation of aldose
reductase and ascorbate peroxidase (APX) enzymes, contributing to the protection against
salinity stress in tomato plants. Additionally, SA treatment led to the accumulation of
osmolytes such as sugars, sugar alcohols, and proline, which function as compatible solutes
and help maintain cellular osmotic balance under saline conditions.

Thus, exogenous SA treatments have been shown to enhance plant growth and seed
germination under NaCl stress. The effectiveness of these treatments depends on various
factors, including SA concentrations, plant species, application mode, plant physiological
state, salinity level, and exposure time to NaCl. The mechanisms underlying the protective
effects of SA involve the enhancement of photosynthetic efficiency, increased activity of
antioxidant enzymes, and the accumulation of osmolytes. Further research is needed to
explore the specific signalling pathways and molecular mechanisms involved in the SA-
mediated responses to salinity stress in different plant species.

27
Overall, the findings presented in this thesis highlight the potential of exogenous SA
treatments as a strategy to improve plant tolerance to NaCl stress. Understanding the intricate
interactions between SA and salt stress response pathways will contribute to the development
of novel approaches for enhancing crop productivity in saline environments.

For instance the soaking of wheat seeds in 0.05 mM SA also reduced the salinity damage on
seedling growth and accelerated the growth processes (Shakirova et al. 2003). The treatment
of wheat plants with SA increased the level of cell division within the apical meristem of
seedling roots, causing an increase in plant growth and elevated wheat productivity
(Shakirova et al. 2003). A high ABA level was also maintained in wheat seedlings, treated
with SA. The SA-induced increase in ABA might contribute to the preadaptation of plants
to stress, since ABA is known to have a key role in inducting the synthesis of a range of
stress proteins ensuring the development of antistress reactions, for example, the maintenance
of proline accumulation (Bandurska and Stroinski 2005). The stress-induced accumulation of
active oxygen species and, therefore, the level of SOD and peroxidase activity in the roots of
young wheat seedlings, pre-treated with SA, were significantly lower than in untreated plants,
indicating that these enzymes contribute to the protective effect of SA on plants under
conditions of salination (Sakhabutdinova et al. 2004). It was also shown that a salt-induced
protein (SALT) was present in the xylem parenchyma cells of vascular bundles in the major
and minor leaf veins. The expression of the gene encoding SALT was upregulated following
the treatment with a fungal elicitor, JA, ABA, or NaCl. However, SA alone or in combination
with one of the other elicitors not only strongly inhibited SALT gene expression but also
exhibited an antagonistic effect in suspension cells and leaves (Kim et al. 2004). The foliar
application 1 mM SA alleviated the NaCl (100 mM) damage in tomato plants and plant
growth was strongly reduced in salt-stressed plants, but this reduction was less pronounced in
SA-treated plants (He and Zhu 2008). SA Fig. 23.3 Some possible ways of SA-induced
oxidative stress tolerance to plants (Hasanuzzaman et al. 2017) 23 Role of Salicylic Acid in
Biotic and Abiotic Stress Tolerance in Plants 541 alleviates NaCl stress by lower levels of
lipid peroxidation and H2O2 accumulation as well as the enhancement of the antioxidant
capacity of plants by increases in CAT, APX, DHAR, and ASC and GSH contents (Lee et al.
2010). SA promotes germination under saline conditions by reducing the NaCl-induced
oxidative damage (Rajjou et al. 2006; Lee et al. 2010). In addition, high SA concentrations
can have a toxic effect on plant growth and development possibly due to its reported effect
inducing ROS accumulation (Rao et al. 1997). Borsani et al. (2001) found that SA enhanced

28
the deleterious effect of NaCl or drought stress in Arabidopsis seedlings by increasing the
rate of ROS generation in photosynthetic tissues. NaCl-adapted tomato cells contained a
lower concentration of SA than an adapted cells. The adaptation process to NaCl was also
related with a higher antioxidative capacity because salt-adapted cells also contained higher
basal levels of APX and GR activities (Molina et al. 2002). In a more recent work, Bastam et
al. (2013) reported that the exogenous application of SA improved the tolerance of pistachio
seedlings to NaCl stress (up to 90 mM NaCl). The SA-treated plants showed lower NaCl-
induced injury symptoms, a better growth rate, and higher chlorophyll contents and
photosynthetic capacity than the non-treated plants. In addition, the foliar application of 0.1
mM SA improved the growth of cotton seedlings in the presence of 100 mM NaCl. The SA-
treated plants displayed better growth and photosynthetic rates and showed low ROS
accumulation (O2 .- and H2O2) and lipid peroxidation that correlated with a significant
enhancement of CAT activity (Liu et al. 2014). Salicylic acid mitigated salinity stress-injury
in Solanum lycopersicum by causing characteristic changes in the expression pattern of GST
gene family members such as SlGSTT2, SlGSTT3, and SlGSTF4 (Csiszár et al. 2014).
Exogenously sourced SA (0.5 mM) was reported to improve salt tolerance in Triticum
aestivum due to an enhanced transcript level of antioxidant genes; GPX1, GPX2, DHAR,
GR, GST1, GST2, MDHAR, and GS, and an increased activity of ascorbate (AsA)-GSH
pathway enzymes (Li et al. 2013). In another instance, SA-mediated restoration of membrane
potential and prevention of salt-induced K+-loss via GORK channel and eventually improved
salinity-tolerance were evinced in A. thaliana (Jayakannan et al. 2013).

29
Fig3:SA effect and tackling of plant stress by various methods(Kaya et al., 2023)

30
Fig.4: Some possible ways of SA-induced oxidative stress tolerance to plants (Hasanuzzaman
et al. 2017).

Drought Stress

The effect of salicylic acid (SA) on drought stress appears to be more consistent compared to
its effect on salt stress. Early reports have indicated that SA treatments can improve the
response to drought stress (Munne-Bosch and Penuelas, 2003; Bechtold et al., 2010; Khokon
et al., 2011; Ying et al., 2013). Overall, the improved drought response induced by SA is
associated with an increase or maintenance of plant growth, Rubisco activity, and
antioxidative capacity.

For instance, SA-treated bayberry plants showed better relative water content (RWC),
photosynthetic rates, and higher catalase (CAT) and superoxide dismutase (SOD) activity, as
well as increased proline content compared to non-treated plants (Ying et al., 2013). In wheat
seedlings under drought stress, SA treatment increased Rubisco and SOD activities, as well
as chlorophyll content, contributing to improved drought tolerance (Singh and Usha, 2003).

31
Habibi (2012) observed that SA application in barley plants enhanced antioxidative defenses
and helped maintain photosynthesis under water stress conditions, thus improving drought
tolerance. Miura et al. (2013) found that drought stress induced the expression of PR-1 and
PR-2, which are typical SA-inducible genes, suggesting a potential role for SA in drought
tolerance. Additionally, the treatment of wheat seedlings with 0.5 mM SA alleviated the
growth inhibition induced by drought and was associated with increased levels of ascorbate
(ASC) and glutathione (GSH), as well as upregulation of GST1, GST2, GR, and MDHAR
genes (Kang et al., 2013).

SA has been suggested to act as an ROS scavenger (Kang et al., 2013). Treatment with SA
increased the activity of ASC-GSH cycle enzymes, SOD, and CAT in maize cultivars that
showed different sensitivities to water stress after a period of water withholding, suggesting
that the ASC-GSH cycle contributes to the removal of hydrogen peroxide (H2O2) generated
during the early phase of water stress (Saruhan et al., 2012). Notably, SA has been shown to
induce stomatal closure in a dose-dependent manner in various plant species, including Vicia
faba, Commelina communis, and Arabidopsis (Mori et al., 2001; Lee, 1998; Khokon et al.,
2011). This SA-induced stomatal closure is dependent on ROS generation, as the application
of antioxidant enzymes such as catalase and SOD can suppress the closure. Moreover, the
stomatal closure induced by SA can be completely inhibited by salicylhydroxamic acid
(SHAM), an inhibitor of cell wall peroxidases, suggesting that SA-induced stomatal closure
occurs through the generation of ROS by these peroxidases (Khokon et al., 2011; Miura et
al., 2013). However, in the case of two maize cultivars, foliar spraying with 1 μM SA
reversed the drought-induced stomatal closure (Saruhan et al., 2012).

Thus the induction of drought tolerance through exogenous application of SA holds

significant practical applications in agriculture, horticulture, and forestry. The beneficial
effects of SA on drought response include improved plant growth, maintenance of
physiological parameters, enhanced antioxidative defenses, and regulation of stomatal
closure. SA acts as an ROS scavenger, enhancing the ASC-GSH cycle and reducing oxidative
stress. The SA-induced stomatal closure, mediated by ROS generated by cell wall
peroxidases, contributes to the regulation of water loss. However, it is important to consider
the concentration and timing of SA application, as they can influence the stomatal response.
These findings highlight the potential of exogenous SA application as a practical strategy for
enhancing drought tolerance in various plant species.

32
Heat Stress

Heat stress is a significant environmental challenge for plants, as high temperatures can
negatively impact their growth and development. However, plants have developed various
mechanisms to adapt and survive under such conditions. One crucial player in this process is
phytohormones, which regulate plant responses to heat stress. In particular, salicylic acid
(SA), a plant hormone known for its involvement in defense responses, has been shown to
play a key role in enhancing heat tolerance in plants.

One of the ways SA helps plants adjust to heat stress is by inducing the synthesis of
osmolytes. Osmolytes are small organic molecules that accumulate in plant cells to maintain
osmotic potential, enabling plants to take in more water. A study by Khan et al. (2013)
demonstrated that SA treatment under heat stress increased the production of proline, an
osmolyte that enhances osmotic potential. This increase in proline allowed the plants to have
a higher water intake, positively influencing stomatal aperture (the opening and closing of
leaf pores) and the photosynthetic machinery. As a result, photosynthesis was more efficient,
with higher rates of photosystem II (PS II) activity and increased Rubisco (a key enzyme in
carbon fixation) activity, leading to improved photosynthetic performance.

Furthermore, SA has been shown to enhance the antioxidant system in plants exposed to heat
stress. Wang and Li (2006) found that SA treatment increased the activity of antioxidative
enzymes, which help mitigate the damage caused by reactive oxygen species (ROS)
accumulation during heat stress. ROS, such as hydrogen peroxide (H2O2), can cause cellular
damage and oxidative stress. By upregulating the antioxidant system, SA helps plants
scavenge ROS and maintain cellular homeostasis.

In addition to its role in osmolyte synthesis and antioxidant defense, SA also influences the
activity of heat shock transcription factors (HSFs) and heat shock proteins (HSPs), which are
crucial components of the plant's heat stress response. SA has been shown to stabilize the
trimers of HSFs and aid their binding to the heat shock element in the promoter of HSP genes
(Jurivich et al., 1992). This activation of HSP genes leads to the synthesis of heat shock
proteins, which assist in protein folding and prevent protein denaturation under heat stress
conditions.

33
Moreover, SA affects ion transport and calcium signalling in response to heat stress. Liu et al.
(2008) suggested that SA induces heat tolerance by altering the activities of plasma
membrane H+ and Ca2+ ATPases. SA treatment increased the activity of H+-ATPase, which
maintains the stability of the plasma membrane, and activated the Ca2+-calmodulin (CaM)
system, leading to an increase in Ca2+ levels in cells. The elevated Ca2+ levels further
stimulated the activity of Ca2+-ATPase, which regulates the cytoplasmic Ca2+
concentration. This regulation helps protect cells and prevent injury caused by heat shock.

Exogenous application of SA has been demonstrated to enhance thermotolerance in various

plant species, including tobacco and Arabidopsis (Lopez-Delgado et al., 1998; Clarke et al.,
2004). For example, spraying grape plants with 0.1 mM SA reduced lipid peroxide levels and
electrolyte leakage in young grape leaves and increased the activity of antioxidative enzymes
(Wang and Li, 2006). Similarly, foliar spraying of cucumber plants with 1 mM SA induced
heat tolerance, as evidenced by lower electrolyte leakage, reduced H2O2 levels, and
increased catalase activity (Shi et al., 2006).

Overall, SA plays a multifaceted role in enhancing heat tolerance in plants. It promotes the
synthesis of osmolytes, boosts the antioxidant defense system, activates heat shock responses,
and modulates ion transport and calcium signalling. Through these mechanisms, SA helps
plants adjust and adapt to heat stress by protecting cellular integrity, maintaining
photosynthetic efficiency, and mitigating oxidative damage. Exogenous application of SA has
proven effective in improving plant thermotolerance, making it a promising approach for
enhancing heat stress resilience in various crops and plant species.

Cold Stress

Plants can be classified as either cold-sensitive or cold-tolerant based on their ability to

withstand low temperatures. Cold-sensitive plants are susceptible to damage or death when
exposed to temperatures ranging from 0 to 15 degrees Celsius, while cold-tolerant plants can
thrive near freezing point and endure temperatures as low as 10-15 degrees Celsius below
zero (Sanghera et al., 2011).

Cold sensitivity in plants is influenced by genetic factors, as well as the stage of development
and metabolic activity. Research indicates that differences in membrane composition play a
significant role in determining sensitivity to low-temperature stress. Cold-sensitive plants

34
have a lower quantity of unsaturated fatty acids compared to cold-tolerant plants (Steponkus
et al., 1993).

To avoid freezing, plants accumulate osmolytes, such as sugars, polyalcohols, amino acids,
polyamines, and quaternary ammonium compounds, in response to low temperatures. This
accumulation lowers the freezing point within the plant cells and prevents cytoplasmic
dehydration caused by frost. Phenol derivatives, including salicylic acid (SA), have been
found to enhance the cold tolerance of plants (Luo et al., 2012).

Studies have demonstrated that treating young maize plants with 0.5 mM SA in the
hydroponic growth solution under normal conditions provides protection against subsequent
low-temperature stress. This observation is supported by measurements of chlorophyll
fluorescence parameters and electrolyte leakage from the leaves (Janda et al., 1997, 1999).
Similarly, pre-treating chilling-sensitive banana seedlings with a 0.5 mM SA solution through
foliage spraying or root irrigation for one day enhances chilling tolerance during subsequent
5 degrees Celsius chilling stress (Kang et al., 2003). SA pre-treatment activates the activities
of superoxide dismutase (SOD), catalase, and ascorbate peroxidase (APX) during the chilling
stress, potentially linked to H2O2 metabolism. Pre-soaking seeds before sowing has also been
shown to improve cold tolerance. In tomato and bean plants, concentrations of 0.1 mM and
0.5 mM SA have been effective against heat, drought, and low-temperature stress (Senaratna
et al., 2000).

Priming pepper seeds (Capsicum annuum L.) with aspirin in KNO3 solution at concentrations
ranging from 0.05 to 0.5 mM enhances germination percentage and improves germination
speed and quality under low-temperature conditions (Korkmaz, 2005). The positive effects of
SA on cold tolerance extend to freezing conditions in winter wheat as well. Exogenous SA
applied at concentrations of 0.01, 0.1, and 1 mM reduces freezing injury in winter wheat
leaves exposed to cold temperatures. Cold conditions increase ice nucleation activity
facilitated by apoplastic proteins in the leaves, and SA treatment further augments this
activity. These results demonstrate that SA can enhance freezing tolerance in winter wheat by
influencing apoplastic proteins. However, it is important to note that this effect may not be
applicable to all plants, as experiments with winter rye revealed that apoplastic proteins
accumulated after SA treatment had no antifreeze activity (Yu et al., 2001).

35
Heavy Metal

The exposure of plants to heavy metals often triggers the overproduction of reactive oxygen
species (ROS) as a prime response. Some metals can directly generate ROS through Haber-
Weiss reactions, while others indirectly disrupt the antioxidative defence system, electron
transport chain, or the metabolism of essential elements (Yadav, 2010). The overproduction
of ROS, including superoxide radicals, hydrogen peroxide, hydroxyl radicals, and singlet
oxygen, is highly toxic and leads to oxidative damage in nucleic acids, proteins,
carbohydrates, and lipid peroxidation (Gill and Tuteja, 2010).

To counteract uncontrolled oxidation, plants have developed an antioxidative defense system,

which consists of both enzymatic and nonenzymatic components. The enzymatic antioxidants
include superoxide dismutase (SOD), guaiacol peroxidase (POD), catalase (CAT),
dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR),
glutathione reductase (GR), ascorbate peroxidase (APEX), glutathione peroxidase (GPX),
and glutathione S-transferase (GST). Nonenzymatic antioxidants encompass glutathione
(GSH), ascorbic acid (ASA), and phenolic compounds such as α-tocopherols, which aid in
scavenging and detoxifying ROS (Gill and Tuteja, 2010; Sharma et al., 2012).

The application of salicylic acid (SA) at a concentration of 0.1 or 0.2 mM has been found to
reduce the inhibitory effects of Pb2+ and Hg2+ on seed germination and early seedling
growth in two rice cultivars (Mishra and Choudhuri, 1997). Additionally, pre-treating barley
seedlings with 0.5 mM SA concentration protected them from cadmium (Cd) toxicity during
subsequent growth periods (Metwally et al., 2003). However, SA treatments strongly or
completely suppressed the Cd-induced upregulation of antioxidant enzyme activities,
suggesting that SA alleviates Cd toxicity through mechanisms other than antioxidant defence.
These mechanisms may involve the binding of Cd, resulting in lower levels of free Cd in the
plasma, or the stimulation of specific ABC transporters involved in the vacuolar sequestering
of Cd products (Eichhorn et al., 2006; Rea et al., 1998).

Nevertheless, SA has been shown to induce the activities of antioxidative enzymes under
metal stress in various studies (Chen et al., 2007; Guo et al., 2007; Wang et al., 2009;
Parashar et al., 2014; Zhang et al., 2015), although some researchers have reported a decline
in catalase (CAT) activity under metal stress (Pandey et al., 2013). Exogenous application of
SA has been found to slightly increase NADH oxidase activity (which stimulates H2O2
generation) in mercury-exposed roots of alfalfa. Moderate levels of H2O2 act as secondary

36
messengers, triggering signalling cascades that activate defence mechanisms under stress
(Zhou et al., 2009).

Some studies suggest that SA, being an iron-chelating molecule, acts as an antioxidant by
directly scavenging hydroxyl radicals, which may lead to a reduction in metal-induced
increases in antioxidative enzyme activity (Ahmad et al., 2011). The enhanced growth
characteristics observed in plants exposed to Cd and sprayed with 10-5 M of SA may be
attributed to the fact that SA acts at the transcriptional and/or translational level, thereby
increasing the activity of various other enzymes necessary for plant growth (Hayat et al.,
2010). Furthermore, SA treatment has been reported to increase crop yield by delaying
senescence in leaves and flowers, extending the duration of photosynthetically active sites
and preventing premature loss of flowers and fruit (Imran et al., 2007).

Salicylic Acid Biosynthesis in Plants

Plants employ two distinct pathways for synthesizing salicylic acid (SA): isochorismate
synthase (ICS) and phenylalanine ammonia-lyase (PAL) (Lefevere et al., 2020). In the ICS
pathway, which primarily occurs in the chloroplasts, chorismate is directly converted into
isochorismate by the enzyme ICS (Murphy et al., 2020). Isochorismate is then transformed
into isochorismoyl-glutamate by isochorismoyl-glutamate synthase, and this compound can
naturally form or be converted into SA through the action of Enhanced Pseudomonas
susceptibility 1 (EPS1), which encodes a BAHD acyl transferase-like protein (Murphy et al.,
2020; Zhong et al., 2021; Kaur et al., 2022a).

On the other hand, the PAL pathway predominantly takes place in the cytoplasm. It begins by
converting phenylalanine into cinnamic acid, followed by the decarboxylation of the side
chain of cinnamic acid to produce benzoic acid. Finally, benzoic acid is converted into SA
through the action of benzoic acid 2-hydroxylase (Zhong et al., 2021; Kaur et al., 2022a). The
involvement of the PAL gene in nematode-attacked wheat plants, with AevPAL1 being
silenced, provides evidence for this pathway. It is suggested that this gene, in conjunction

37
with AevTDC1, alters the content of SA and secondary metabolites, thereby enhancing
resistance to cereal cyst nematodes in Aegilops variabilis (Zhang et al., 2021). Different plant
species exhibit varying degrees of dependence on these mechanisms for SA production
(Tripathi et al., 2019). Arabidopsis relies more on the ICS pathway, while the PAL pathway
seems to be more prevalent for SA formation in rice (Lefevere and Gheysen, 2019).

Once synthesized, SA undergoes various physiologically significant chemical modifications,

including glucosylation, methylation, and conjugation with amino acids (AA), which render
SA inactive (Mishra and Baek, 2021). The glucosylation of SA results in the formation of
SA-glucoside (SAG), a product that can be stored in significant quantities in the vacuole (Li
et al., 2021). The reverse process of glucosylation may be facilitated by unidentified glucose
hydrolases or β-glucosidases (Ding and Ding, 2020). Glucosyltransferases, such as UGT74F1
and UGT76B1, utilize the hydroxyl group of SA as a substrate for the formation of glucosyl
salicylate (GS) (Mohnike et al., 2021). Both SAG and GS can accumulate in tobacco plants
during defense responses against Pseudomonas syringae pv. phaseolicola, indicating their
association with plant defense (Lee and Raskin, 1998). Notably, the levels of free SA and
SAG increase concurrently during plant defense reactions (Nobuta et al., 2007). Studies have
shown that exogenous application of SA is sufficient to restore SAG accumulation, PR1
expression, and improved resistance to virulent pathogens. The PBS3 gene is vital for SA
metabolism, as mutations in PBS3 affect SA-dependent gene expression, SAG accumulation,
and disease resistance. Salicyloyl glucose ester (SGE), another glucosylated SA sugar
molecule, may also be present in plants and is involved in controlling plant disease resistance
(Berim and Gang, 2022). Silencing of the CsUGT87E7 gene in tea plants leads to a reduction
in SGE levels and increased susceptibility to pathogens (Hu et al., 2022).

SA becomes more mobile following its conversion into methyl salicylate (MeSA).
Methylation enhances its membrane permeability (Saleem et al., 2021a). MeSA is released by
numerous plants in response to insect herbivore feeding and acts as a component of
herbivore-induced plant volatile blends, mediating interactions between plants and insects
(Filgueiras et al., 2019). Pathogen attack significantly stimulates MeSA production, and its
volatile nature allows it to act as a systemic acquired resistance (SAR) signal, transmitting
through the phloem and other tissues to various parts of the plant (Kachroo and Kachroo,
2020). MeSA has been found to enhance the production of phenolic antifungicides, such as
chlorogenic acid, catechin, and phloridzin, by stimulating specific enzymes in the
phenylpropanoid pathway (Gacnik et al., 2021). The contribution of MeSA to SAR mobile

38
signals, however, is still debated, as more recent research has indicated that initial infection is
required for SAR formation (Kachroo et al., 2022).

Following synthesis, SA can undergo various conversions, leading to the formation of

functional or dysfunctional compounds (Ding and Ding, 2020). Conjugation with amino acids
is an essential type of modification, involving the attachment of carboxylic acids to hormones
through the formation of amide bonds (Westfall et al., 2016). For instance, aspartic acid
(Asp) can be conjugated to SA, forming SA-Asp, an inactive form of SA. This process is
catalyzed by homologs of the glycoside hydrolase 3 (GH3) protein family (Tiwari et al.,
2022). SA-Asp is the most prevalent permanent conjugate found in plant species such as
Arabidopsis, Cucumis sativus, and Selaginella moellendorffii (Chen et al., 2013; Dong et al.,
2020; Berim and Gang, 2022). Limited information is available regarding SA-amino acid
conjugates and their physiological roles, and it remains to be verified whether they are
formed through alternative pathways or represent a biological output of SA.

Fig.5: ICS and PAL pathway of SA synthesis (Kaya et al., 2023)

39
Fig:6: Schematic outline of phenylalanine pathway in SA biosynthesis (PAL phenylalanine
ammonia lyase, BA2H benzoic acid 2-hydroxylase) (Lee et al. 1995).

Future Perspectives

Salicylic acid (SA) is a crucial signalling molecule in plants, playing a significant role in
various physiological processes, including plant growth and stress responses. Its involvement
in plant defence against biotic and abiotic stresses has been extensively studied. As we move
forward, deeper exploration of SA's mechanisms of action and its crosstalk with other
signalling pathways will open up exciting future perspectives in understanding and
harnessing its potential for sustainable agriculture and stress tolerance in plants.

Unravelling the Signalling Pathways: Future research will focus on unravelling the intricate
signalling pathways regulated by SA. Understanding the downstream targets and interaction
with other phytohormones, such as jasmonic acid and ethylene, will provide comprehensive
insights into the complex network of stress responses. This knowledge will enable scientists
to fine-tune SA signalling for tailored stress management strategies in crop plants.

Engineering Stress-Tolerant Crops: Advancements in biotechnology and genome editing

techniques will facilitate the engineering of stress-tolerant crops by manipulating SA-related

40
pathways. Precise gene editing or transgenic approaches can be employed to enhance SA
biosynthesis, perception, or downstream responses. Engineered crops will exhibit improved
resistance to pests, diseases, and environmental stresses, ultimately leading to increased
agricultural productivity and food security.

Enhancing Plant Growth and Development: Further investigations into the role of SA in
plant growth and development will unlock its potential as a growth regulator. Understanding
how SA influences key processes, such as seed germination, root development, flowering,
and fruiting, will provide opportunities to optimize crop growth and yield. The application of
SA as a growth-promoting agent will be refined, considering dose-specific responses and
environmental contexts.

Exploring Crosstalk with Beneficial Microbes: The interaction between SA and beneficial
microbes in the rhizosphere will be explored for sustainable agriculture. Understanding how
SA levels are modulated by beneficial microbes and how SA reciprocally influences
microbial activity will aid in designing bio-fertilizers and biocontrol agents that can enhance
plant growth and defense responses. Harnessing these interactions will promote a balanced
and resilient soil ecosystem.

SA in Climate Change Mitigation: As climate change poses increasing challenges to

agriculture, future research will investigate the role of SA in mitigating climate-induced
stresses. Studying how SA can alleviate the adverse effects of heat, drought, and salinity
stresses will be critical. Utilizing SA to improve plant water use efficiency and
thermotolerance will be crucial for adapting agriculture to changing climatic conditions.

The future perspectives on SA with respect to plant growth and stress response hold
immense promise for sustainable agriculture and global food security. Advances in
understanding SA signalling, gene editing technologies, and beneficial microbe interactions
will pave the way for innovative approaches to enhance crop stress tolerance and
productivity. The potential applications of SA in climate change mitigation and growth
regulation further underscore its significance in shaping the future of agriculture.

Potential mechanisms to mitigate the negative effects of stress through SA include :

(a) SA collaborates with other plant hormones and signaling molecules, participating in
intricate signaling pathways. (b) SA induces the production of H2O2, acting as a secondary
messenger that triggers diverse reactions to counteract abiotic stressors, and indirectly

41
influences the antioxidant status. (c) SA scavenges free radicals, thereby safeguarding the
stability of organelles and cell membranes. (d) In cases of heavy metal (HM) toxicity, SA
increases the levels of GSH in HM-stressed seedlings by regulating HM sequestration to
render them nontoxic.

Despite these advances, gaining a comprehensive understanding of hormonal interactions,

especially relating to SA's role in plant growth, development, and stress responses, remains
challenging. The intricate relationship between SA and other plant hormones, along with their
various metabolic impacts, presents complexities in plant regulation. This review focuses on
the most recent research on these inter-relationships and consolidates prior knowledge about
the functions of SA cross-talk in plant metabolism.

To further our understanding of SA's multiple roles in plants, more studies are needed to
uncover the genetic network of SA in the future. By utilizing SA-deficient or SA-
overproducing mutants under stress conditions, researchers can delve deeper into the
crosstalk between SA and other phytohormones and signaling molecules. This approach
promises to shed light on the complex interactions that govern plant responses to their
environment and aid in the development of strategies to enhance crop growth and stress
tolerance.

Conclusion:

Salicylic acid (SA) plays a vital role in plant growth, development, and stress response, with
a particular emphasis on stress management. Through extensive research, it has become
evident that SA functions as a key signalling molecule, orchestrating various defence
mechanisms against biotic and abiotic stresses. As we delve deeper into the understanding of
SA's molecular mechanisms and its intricate crosstalk with other signalling pathways, its
potential for mitigating stress and promoting plant growth becomes increasingly apparent.

42
The signalling pathways regulated by SA have been subject to extensive investigation, with
researchers striving to unravel the complex network of downstream targets and interactions
with other phytohormones. Further exploration of these pathways will provide valuable
insights into the intricacies of stress response mechanisms in plants. This knowledge will
enable scientists to develop tailored stress management strategies that utilize SA as a key
component.

One exciting avenue for future research lies in the engineering of stress-tolerant crops by
manipulating SA-related pathways. With the advent of advanced biotechnological tools and
genome editing techniques, it becomes possible to precisely enhance SA biosynthesis,
perception, or downstream responses in crops. By doing so, we can develop plants that
exhibit heightened resistance to pests, diseases, and environmental stresses, thereby
increasing agricultural productivity and ensuring food security.

In addition to stress mitigation, SA also influences various aspects of plant growth and
development. Research efforts are directed towards understanding how SA influences seed
germination, root development, flowering, and fruiting processes. By elucidating these
mechanisms, we can optimize the use of SA as a growth-promoting agent in crops, taking
into account dose-specific responses and environmental contexts.

Moreover, the interaction between SA and beneficial microbes in the rhizosphere represents a
promising area for further exploration. The modulation of SA levels by beneficial microbes,
as well as the reciprocal influence of SA on microbial activity, can be harnessed to design
bio-fertilizers and biocontrol agents that enhance plant growth and defence responses.
Understanding and harnessing these interactions will foster a balanced and resilient soil
ecosystem.

As climate change poses increasing challenges to agriculture, the potential of SA in

mitigating climate-induced stresses gains significance. Investigations into the role of SA in
alleviating the adverse effects of heat, drought, and salinity stresses will be critical. Utilizing
SA to improve plant water use efficiency and thermotolerance will be crucial for adapting
agriculture to changing climatic conditions and ensuring sustainable food production.

In conclusion, the extensive research conducted on SA with respect to plant growth,

development, and stress highlights its central role in plant physiology. Future perspectives in
this field offer immense potential for sustainable agriculture and global food security..

43
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