Owing to its lipophilic and highly diffusible in nature, NO is involved in several signaling processes in plants
[107[105]]. The list of NO-derivatives, usually referred to as RNS includes NO radicals (NO
−), nitrosonium cation (NO
+), peroxynitrite (ONOO
−), S-nitrosothiols (SNOs), higher oxides of N (NOx) and dinitrosyl-iron complexes
[108[106]]. S-nitrosoglutathione (GSNO) is yielded as a result of the reaction of NO with reduced GSH in presence of O
2. In turn, NADH-dependent S-nitrosoglutathione reductase (GSNOR) decomposes GSNO to GSSG and NH
3 [109 [107]]. NO synthesized during these processes can induce diverse effects in plants
[110[108]]. Conflicting reports are accessible about NO induced changes, where some studies show NO as a stress inducing agent
[111[109]], even as others have mentioned it as a protective agent
[17[17]]. In order to play signaling function, NO has been shown to interact with second messenger, such as, cyclic guanosine monophosphate (cGMP), cytosolic Ca
2+ ion concentration, and cADP-ribose, phosphatidic acid and ROS
[112[110]]. NO-mediated regulation of the activity of enzymes may involve post-translational modifications and joining themselves with metal centers of proteins. NO may also alter the aggregation state, activity, or location of residue of Cys and tyrosine of proteins
[99[97]]. NO-mediated nitration of fatty acids is an imperative part of NO signaling in plants
[113[111]]. NO can also initiate mitogen-activated protein kinase (MAPK) signaling pathways in plant cells
[114[112]]. MAPK cascade (MEK2-SIPK) induce
s NO ASSOCIATED1 and NR-mediated NO burst
[115[113]]. In particular, SIPK can give positive feedback between NO and ROS signals and MAPK cascades can act to regulate ROS and NO production in plants
[115[113]]. Reduction in ROS levels, mitigation of oxidative stress and the role as an anti-senescence agent can be performed by the mitochondrial NOS (
NOS1).
6.2. Nitric Oxide in Salt Tolerance
The literature supports the involvement of NO in plant stress-response mechanisms, which are largely contradictory, and are contingent to plant species types and the extent of the salt treatment. On the one hand, NO-mediated improvement in plant health was reported to involve improved osmolytes accumulation and strengthened antioxidative defense systems
[116[114]]. NO can also mitigate H
+-ATPase inhibition, enhance mineral absorption and maintain hormone equilibrium; and thereby attenuate growth inhibition
[117[115]]. The NO-melatonin interaction regulates the expression of Cu/Zn-SOD and Mn-SOD, eventually maintains oxidative homeostasis in plants under salt stress
[118[116]]. Moreover, the family of protein kinase is also involved in NO-mediated signaling cascades in salt stressed plants. Furthermore, comparative study of NO and SA was done to study their impact on salt-stress mitigation. The results speculated that NO enhanced salt tolerance due to antioxidant enzymes and compatible solute. However, SA was effective to reduce MDA and H
2O
2 content
[119[117]]. The supplementation of NO and jasmonic acid (JA) was reported to mitigate salt stress toxicity through modulation of antioxidant enzymes
[96[94]]. NO-interaction with phytohormones can be useful in plant salinity tolerance. I
n Arabidopsis, abiotic stress can reduce root meristem growth by decreasing accumulation of auxin-mediated by NO
[120[118]]. The auxin-mediated initiation of NADPH-dependent thioredoxin reductase (NTR) activity is reduced by NO scavengers, signifies that NO is downstream of auxin in this regulatory pathway. NO was observed to efficiently reduce the inhibitory effects of salt stress on growth rates and exosmosis of electrolytes and decreases the lipid peroxidation in wheat as a result of increment in cytokinin (CK)
[121[119]]. Furthermore, the supply of absicsic acid (ABA) improves the production and participation of NO downstream of ABA and thereby controls processes including seed germination, dormancy and stomatal movement
[122[120]].
7. Ethylene
First identified as a simple gaseous molecule (28.054 g mol
−1) ethylene (ETH; CH
2=CH
2), is produced in all higher plants and usually connected with fruit ripening and the triple response. ETH has been reported first as the active component in illuminating gas caused a triple response in etiolated pea seedlings
[123[121]]. The development o
f A. thaliana as a model system and eventual development and screening of genetic mutants largely contributed in ETH signaling
[124[122]]. The extension in our understanding on how this phytohormone works was possible as a result of the identification of gene array and transcriptional factors working downstream
[125[123]].
Ethylene regulates diverse aspects of plant growth responses and development
[126[124]] and also plays very important roles in photosynthesis under abiotic stress
[22[22]] by regulation of stomatal movement and more influx of CO
2 important for carboxylation
[127[125]]. The list of major factors involved in the potential control of ETH-production includes internal signals throughout the development and plant responses under varied abiotic (nutritional disorders, wounding, hypoxia, ozone, chilling, or freezing) and biotic (e.g., pathogen attack) stresses
[128[126]]. Furthermore, ETH can regulate H
+-ATPase gene expression and thereby control ion homeostasis in salt stressed plants
[129[127]].
7.1. Ethylene Biosynthesis and Signaling
The ETH biosynthetic pathway in higher plants is well described
[130,131,132,133[128][129][130][131]]. ETH-production is strictly regulated at various levels and involves both transcriptional
[134[132]], post-translational regulation
[135[133]]. ETH-signaling cascade starts with first perceived by a family of membrane-bound receptors, predominantly localized at the endoplasmic reticulum (ER). I
n Arabidopsis, ETH-receptors (
ETHYLENE RECEPTOR1 an
d 2 (ETR1/2)
, ETHYLENE RESPONSE SENSOR 1 an
d 2 (ERS1/2) an
d ETHYLENE INSENSITIVE 4 (EIN4) perceive ETH and also involves a number processes
[136[134]]. In particular, EIN3 and its close homolog
, ETHYLENE INSENSITIVE3-LIKE1 (EIL1) are key transcription factors activated by EIN2
[137[135]].
7.2. Ethylene in Salt Tolerance
ETH signaling contributes in plant stress responses and tolerance to abiotic stresses including salinity
[16,22,138[16][22][136]]. ETH can maintain the homeostasis of ions and also up-regulate antioxidant enzymes in plants under environmental fluctuation
[139[137]]. Exogenous application of ACC lowered osmotic stress and controlled non-photochemical quenching (NPQ) via PSI cyclic electron flow and starch metabolism in tomatoes under salt stress
[21[21]]. MAPK cascade-mediated control of the activation of different defense mechanisms also contributes in response to ROS stress
[140[138]].
The supply of ETH-releasing compounds (such as ethephon), ETH-precursors (such as ACC) and/or endogenous ETH-overproduction were reported to confer plant salt tolerance
[141,142[139][140]]. ETH may act as both a positive regulator and or a negative regulator in salt stress-resistance in plants. To this end, application of 1-MCP, an ETH-action inhibitor improved physiological and biochemical antioxidant characteristics
[46[44]]. Calcium carbide (CaC
2), a precursor of ETH was reported to exhibit its ability to alleviate salt stress and improve ETH-concentration, seed germination, osmolytes and activities of α-amylase and decrease H
2O
2 and MDA contents under salt stress
[143[141]]. Ethylene contribution to salt acclimation processes can differ with respect to the type of response, including improved expression of ETH receptors and ETH production
[144[142]]. However, ETH involvement in plant response to salt resistance is highly complex and still remains unclear. In addition, EIN3/EILs are the key elements and positive factors in ETH-mediated downstream transcriptional cascade
[145[143]]. EIN3/EILs are involved in many physiological processes, seed germination, apical hook formation, fruit development, abiotic stress and hormone responses
[145,146[143][144]]. A recent report indicates that EIN3/EIL1 are necessary for the enhanced ETH-induced salt tolerance i
n Arabidopsis and salt stress leads to a large number of EIN3/EIL1-regulated genes via promoting EBF1/EBF2 proteasomal degradation in EIN2-dependent and EIN2-independent pathways
[147[145]]. Overexpression of EIN3 target genes (e.g., ERFs an
d SALT INDUCED EIN3/EIL1-DEPENDENT1 (SIED1)) in a range of plant species leads to salt tolerance
[144[142]]. Moreover, melatonin was also reported to enhance plant salt-tolerance by promotin
g MYB108A-mediated ETH biosynthesis
[148[146]].
Signaling molecules such as ABA, CK, indole acetic acid (IAA), gibberrelins (GA), ROS, and Ca
2+ interacts with ETH and thereby contribute in plant stress-adaptation
[15,138,149,150[15][136][147][148]]. The DELLA proteins and GA-opposable growth inhibitors of the GA signaling pathway participate in plant salt-tolerance
[151[149]]. In particular, ETH-mediated increase in the accumulation of DELLA proteins reduced the action of GA
[138[136]]. ABA was reported to regulate the ETH biosynthesis-associated genes (
AtACS5, AtACS7, TaACO1, an
d GhERF1) under salt stress
[138[136]]. ETH and JA may act synergistically or antagonistically in response to stressed conditions and mediate via EIN3/EIL1 along wit
h JAZs-MYC2 [152 [150]].
The relationship between ETH and SA are antagonistic, conferring tolerance against environmental fluctuation
[153[151]]. The application of SA restricted the ETH production by inhibiting the activity of ACC synthase under salt stress
[154[152]]. As a result, the reduction in the synthesis of ETH alleviated the toxicity of salt stress. Nazar et al.
[155[153]] suggested that SA treatment constrained the ETH formation by inhibiting the ACC synthase activity under drought stress. Interactive outcomes of ETH, H
2O
2, and brassinosteroids were reported to confer plant-tolerance against salt stress
[156[154]].
8. Crosstalk of NO with N and S in Salt Tolerance
The NO generation and assimilation of N are closely connected. Both the amount and also the form of the supplied N (i.e., NO
3− or/NH
4+) affect levels of NO in plants
[157[155]]. Nitrite (NO
2−) and arginine are the main substrates for NO synthesis. Plants were reported to optimize the use of nitrite as a main source for NO
[158[156]]. NO-regimes were reported to exaggerate the activity NR, a key enzyme in N-metabolism, and a NO-source
[159[157]]. NO was also argued to modulate the uptake and distribution systems of N
[160[158]]. In addition to its role in N assimilation and uptake, involvement of NO in nitrate-sensing pathway was also reported in plants
[161[159]]. The reaction of NO with GSH produces GSNO, a low-molecular-weight S-nitrosothiol (SNO). Notably, more stable than NO, SNO serves as a mobile reservoir of NO. However, GSH content can also regulate plant’s S-status through induced S assimilation and forms GSNO
[17,162[17][160]]. The coordination of NO with S can maintain ionic and redox homeostasis, strengthen the plant antioxidant machinery, and thereby improve photosynthetic performance, growth, and salt tolerance
[17[17]]
. Figure 2 summarizes N and S assimilation and how their coordination with NO helps in salt tolerance in plants.