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Verticillium wilt (VW), a fungal disease caused by Verticillium dahliae , currently devastates cotton fiber yield and quality seriously, yet few resistance germplasm resources have been discovered in Gossypium hirsutum . The cotton variety Nongda601 with suitable VW resistance and high yield was developed in our lab, which supplied elite resources for discovering resistant genes. Early nodulin-like protein (ENODL) is mainly related to nodule formation, and its role in regulating defense response has been seldom studied. Here, 41 conserved ENODLs in G. hirsutum were identified and characterized, which could divide into four subgroups. We found that GhENODL6 was upregulated under V. dahliae stress and hormonal signal and displayed higher transcript levels in resistant cottons than the susceptible. The GhENODL6 was proved to positively regulate VW resistance via overexpression and gene silencing experiments. Overexpression of GhENODL6 significantly enhanced the expressions of salicylic acid (SA) hormone-related transcription factors and pathogenicity-related (PR) protein genes, as well as hydrogen peroxide (H 2 O 2 ) and SA contents, resulting in improved VW resistance in transgenic Arabidopsis . Correspondingly, in the GhENODL6 silenced cotton, the expression levels of both phenylalanine ammonia lyase (PAL) and 4-coumarate-CoA ligase (4CL) genes significantly decreased, leading to the reduced SA content mediating by the phenylalanine ammonia lyase pathway. Taken together, GhENODL6 played a crucial role in VW resistance by inducing SA signaling pathway and regulating the production of reactive oxygen species (ROS). These findings broaden our understanding of the biological roles of GhENODL and the molecular mechanisms underlying cotton disease resistance.

Background: Increased protein carbonylation is a hallmark of oxidative stress, protein homeostasis dysregulation and aging in the nervous system and skin. Sensory neurons interact with skin cells and are involved in skin homeostasis. We have previously reported that the 5-octanoyl salicylic acid (C8-SA), a salicylic acid derivative, increased C. elegans lifespan and delayed the accumulation of carbonylated proteins, through the stimulation of autophagy., Objectives: In this study we aimed to investigate if C8-SA protects human sensory neurons and human skin from extrinsic oxidative stressors as an approach to delay skin aging., Methods: In vitro reconstituted human epidermis innervated with hiPSc-derived human sensory neurons, as well as ex vivo human organotypic full skin models were used. The fully differentiated sensory neurons were pretreated with C8-SA before oxidative stress induction. Skin explants were maintained in culture and treated topically with C8-SA before the application of urban pollutants. Carbonylated proteins were detected using amino-oxy functionalized fluorophores and quantified. Chaperone mediated autophagy was monitored with LAMP2A immunofluorescence. Inflammation, ROS detoxification and autophagy were assessed by RT-PCR., Results: C8-SA prevented the accumulation of carbonylated proteins, both in human sensory neurons and skin explants. C8-SA stimulated chaperone-mediated autophagy and modulated NRF2 antioxidant response genes, as well as catalase enzymatic activity., Conclusions: C8-SA acts at two levels to protect skin against oxidative stress: 1) it prevents protein oxidation by stimulating endogenous antioxidant defense and 2) it increases the clearance of oxidized proteins by stimulating chaperone-mediated autophagy. These results suggest that C8-SA maintains skin health in urban polluted environments., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Fusarium oxysporum causes significant economic losses in many crop plants by causing root rot, necrosis, and wilting symptoms. Homology and molecular dynamics studies are promising tools for the detection in F. oxysporum of the systemic resistance compound, salicylic acid, for control of the SKP1-CUL1-F-box protein complex. The structure of SKP1-CUL1-F-box subunit Skp1 from F. oxysporum is produced by Modeler 9v7 for the conduct of docking studies. The Skp1 structure is based on the yeast Cdc4/Skp1 (PDB ID: 3MKS A) crystal structure collected by the Protein data bank. Applying molecular dynamic model simulation methods to the final predicted structure and further evaluated by 3D and PROCHECK test programmers, the final model is verified to be accurate. Applying GOLD 3.0.1, SCF Complex Skp1 is used to prevent stress-tolerant operation. The SKP1-CUL1-F-box model is predicted to be stabilized and tested as a stable docking structure. The predicted model of the SCF structure has been stabilized and confirmed to be a reliable structure for docking studies. The results indicated that GLN8, LYS9, VAL10, TRP11, GLU48, ASN49 in SCF complex are important determinant residues in binding as they have strong hydrogen bonding with salicylic acid, which showed best docking results with SKP1-CUL1-F-box complex subunit Skp1 with docking score 25.25KJ/mol. Insilco studies have been used to determine the mode of action of salicylic acid for Fusarium control. Salicylic acid hinders the SKP1-CUL1-F-box complex, which is important in protein-like interactions through hydrogen bodings. Results from docking studies have shown that the best energy for SKP1-CUL1-F-box was salicylic acid.Communicated by Ramaswamy H. Sarma.

Dark green islands (DGIs) are the outcome of post-transcriptional gene silencing (PTGS) in antiviral immunity, but their characteristics related to PTGS remain largely unknown. In this study, the cucumber mosaic virus (CMV) was inoculated on Nicotiana tabacum plants to explore the PTGS features of DGIs. Our results showed that higher expressions of PTGS-associated genes, especially NtAGO1, present in DGIs. To investigate the role of NtAGO1 in the generation and the antiviral effect of DGIs, NtAGO1 was then over-expressed or knocked out in N. tabacum plants through agrobacterium-mediated genetic transformation. The results showed that more DGIs with larger areas appeared on NtAGO1 over-expressed plants, accompanied by less virus accumulation, less reactive oxygen species production, and seldom membrane damage, whereas fewer DGIs appeared on NtAGO1 knockout plants with more damage on infected plants. In addition, the NtAGO1-participated antiviral process could promote the transduction of the salicylic acid-mediated defense pathway. Taken together, our results indicate that DGIs are maintained by a stronger PTGS mechanism, and NtAGO1 positively regulates the generation and viral resistance of DGIs in N. tabacum., (Copyright © 2022. Published by Elsevier Masson SAS.)

Attacks of necrotrophic and biotrophic fungi affect a large number of crops worldwide and are difficult to control with fungicides due to their genetic plasticity. Encapsulation technology is a good alternative for controlling fungal diseases. In this work, encapsulated samples of salicylic acid (SA) with silica (Si:SA) or chitosan (Ch:SA) at three different ratios were prepared by spray drying, and morphological and physicochemical characterised. Therefore, size distribution, specific surface area, thermal stability, encapsulation efficiency, and in-vitro SA release were determined. Biological activity of encapsulated samples were tested against different fungi of agricultural interest at various concentrations (0-1000 µM). Treatments prepared with the lowest ratios for both capsules, were found to have the best antifungal effect in an in vitro system, inhibiting the mycelial growth of Alternaria alternata, Botrytis cinerea, Fusarium oxysporum and Geotrichum candidum. Similarly, treatments with the lowest ratios of both encapsulated samples reduced free SA toxicity on Arabidopsis thaliana seeds. In this system, plants treated with capsules had higher root and rosette development than those treated with free SA. In conclusion, a product with a great potential in agriculture that shows high antifungal capacity and low toxicity for plants have been developed through a controlled and industrially viable process., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Background: Helicteres angustifolia has long been used in Chinese traditional medicine. It has multiple pharmacological benefits, including anti-inflammatory, anti-viral and anti-tumor effects. Its main active chemicals include betulinic acid, oleanolic acid, helicteric acid, helicterilic acid, and other triterpenoid saponins. It is worth noting that some acylated triterpenoids, such as helicteric acid and helicterilic acid, are characteristic components of Helicteres and are relatively rare among other plants. However, reliance on natural plants as the only sources of these is not enough to meet the market requirement. Therefore, the engineering of its metabolic pathway is of high research value for enhancing the production of secondary metabolites. Unfortunately, there are few studies on the biosynthetic pathways of triterpenoids in H. angustifolia, hindering its further investigation., Results: Here, the RNAs of different groups treated by metabolic stimulation were sequenced with an Illumina high-throughput sequencing platform, resulting in 121 gigabases of data. A total of 424,824 unigenes were obtained after the trimming and assembly of the raw data, and 22,430 unigenes were determined to be differentially expressed. In addition, three oxidosqualene cyclases (OSCs) and four Cytochrome P450 (CYP450s) were screened, of which one OSC (HaOSC1) and one CYP450 (HaCYPi3) achieved functional verification, suggesting that they could catalyze the production of lupeol and oleanolic acid, respectively., Conclusion: In general, the transcriptomic data of H. angustifolia was first reported and analyzed to study functional genes. Three OSCs, four CYP450s and three acyltransferases were screened out as candidate genes to perform further functional verification, which demonstrated that HaOSC1 and HaCYPi3 encode for lupeol synthase and β-amyrin oxidase, which produce corresponding products of lupeol and oleanolic acid, respectively. Their successful identification revealed pivotal steps in the biosynthesis of acylated triterpenoids precursors, which laid a foundation for further study on acylated triterpenoids. Overall, these results shed light on the regulation of acylated triterpenoids biosynthesis., (© 2022. The Author(s).)

Phytoplasmas are economically important plant pathogenic bacterial diseases, causing severe yield losses worldwide. In this study, we tested nanoformulations such as glycyrrhizic acid ammonium salt (GAS), salicylic acid (SA), and boric acid (BA) as novel antimicrobial agents inducing the resistance against the phytoplasma disease in faba bean. The nanoparticles (NP) were foliar-applied to naturally phytoplasma-infected faba bean with three concentrations from each of SA, GAS, and BA, under field conditions. Nested PCR (using universal primer pairs P1/P7 and R16F2n/R16R2) were reacted positively with all symptomatic samples and gave a product size of approximately 1200 bp, while the healthy plant gave no results. Transmission electron microscopy examinations of phytoplasma-infected faba bean plants treated with different nanoparticles revealed that severe damage occurred in phytoplasma particle's structure, degradation, malformation, lysis in the cell membrane, and the cytoplasmic leakage followed by complete lysis of phytoplasma cells. Exogenous application of GAS-NP (1.68 µM), SA-NP (0.28 µM), and BA-NP (0.124 µM) suppressed the infection percentage of phytoplasma by 75%, 50%, and 20%, and the disease severity by 84%, 64%, and 54%, respectively. Foliar application of nanoparticles improved Fv/Fm (maximum quantum efficiency of PSII Photochemistry), PI (the performance index), SPAD chlorophyll (the relative chlorophyll content), shoots height, and leaves number, thus inducing recovery of the plant biomass and green pods yield. The most effective treatment was GAS-NP at 1.68 µM that mediated substantial increases in the shoots' fresh weight, shoots' dry weight, number of pods per plant, and green pods yield by 230%, 244%, 202% and 178%, respectively, compared to those of infected plants not sprayed with nanoparticles. This study demonstrated the utility of using nanoparticles, particularly GAS-NP at 1.68 µM to suppress the phytoplasma infection.

Background: The Lamiaceae family, one of the most important herbaceous and shrub plant families, includes a wide variety of plants with biological and medicinal uses. This study aimed to conduct a comparative analysis of phenolic compounds content and biological activity of extracts from eight species of Lamiacea e plants, cultivated in a temperate climate, and to study the effect of the foliar salicylic acid application on these parameters., Methods: Lamiaceae plants ( Lavandula angustifolia , Salvia officinalis , Hyssopus officinalis , Agastache foeniculum , Thymus serpyllum , Mentha × piperita , Origanum vulgare , Monarda didyma ) cultivated on field experimental sites. Plants were sprayed with salicylic acid at a concentration of 1 mM. The untreated with salicylic acid plants were used as control., Results: The highest contents of hydroxycinnamic acids and flavonoids in control plants were observed in the Agastache foeniculum (6.4 ± 0.6 mg g-1 and 6.5 ± 0.4 mg g-1 respectively). The highest content of phenolic compounds was found in Monarda didyma (13.8 ± 0.7 mg g-1). Among the control plants, Agastache foeniculum , Hyssopus officinalis, and Mentha × piperita were characterized by the highest antioxidant activity. All the studied extracts had the ability to inhibit α-amylase and α-glucosidase. Significant positive correlations were revealed between the antioxidant activity and the contents of hydroxycinnamic acids, total phenolic compounds, and flavonoids. A high degree of correlation was found between the α-amylase inhibitory activity and the content of hydroxycinnamic acids ( r = 0.72, p ≤ 0.05), as well as between the α-glucosidase inhibitory activity and the content of flavonoids ( r = 0.83, p ≤ 0.05) and hydroxycinnamic acids ( r = 0.81, p ≤ 0.05). The foliar treatment with salicylic acid led to an increase in the contents of hydroxycinnamic acids (in 6 species), flavonoids (in 2 species), total phenolic compounds (in 7 species), antioxidant activity (in 5 species), as well as in α-amylase (in 4 species) and α-glucosidase (in 5 species) inhibitory activity compared to the control plants. Lavandula angustifolia was the most susceptible to foliar treatment with salicylic acid. In this plant species, a significant increase in all studied biochemical parameters was noted., Conclusions: The results obtained on the stimulating effects of salicylic acid can be used to increase the nutritional and pharmacological value of plants of the Lamiaceae family cultivated in temperate climates., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s). Published by IMR Press.)

Objectives: A new nitronyl nitroxide radical with a salicylic acid framework (SANR) has been demonstrated to exert antioxidant effects in the previous study by our team. The current study has assessed the protective effect of SANR on cerebral ischemia and reperfusion (I/R) in rat models., Methods: Sprague-Dawley rats were randomly divided into four groups: sham, I/R, 10, and 20 mg/kg SANR + I/R groups. A total of 120 min of middle cerebral artery occlusion (MCAO) caused cerebral ischemia. Survival rates were calculated, and neurological deficits were evaluated by a blinded experimenter. Cerebral infarct area, apoptosis cells, and blood-brain barrier (BBB) leakage were measured by 2,3,5-triphenyltetrazolium chloride staining, terminal deoxynucleotidyl transferase dUTP nick-end labeling, and Evans blue assay, respectively. Reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and 8-hydroxy-2-deoxyguanosine (8-OHdG) also were detected to assess oxidation damage caused by cerebral I/R., Results: Treatment with SANR significantly promoted survival of rats with cerebral I/R injury. SANR meliorated neurologic deficit and infarct area, improved BBB permeability, and reduced neuronal apoptosis. SANR also reduced ROS levels and the content of MDA and increased SOD and GSH-Px activity in a dose-dependent manner. Furthermore, SANR could inhibit the expression of 8-OHdG., Conclusion: Our results suggested that SANR has a neuroprotective effect against cerebral I/R injury, and its effect mechanism is related to the antioxidant function., (Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc.)

Background: Rice blast disease (Magnaporthe oryzae) is considered the most destructive rice disease all over the world. Dimetindene maleate is used in medication against allergic reactions in humans. Dimetindene maleate used to induce systemic acquired resistance (SAR) in rice (Oryza sativa L.) in order to protect rice plants from blast disease., Results: Dimetindene maleate was not effective against fungus linear growth in vitro. In glasshouse conditions, dimetindene maleate significantly improved resistance at 25, 50, 125, 250, 500 and 1000 mg L -1 concentrations. Leaf blast severity reached 14.18% on plants treated with the most effective concentration of 125 mg L -1 compared with control plants. In field conditions during both seasons (2016 and 2017), 125 mg L -1 dimetindene maleate decreased the disease severity to 1.1% and 2.7%, respectively, after 30 days of treatment. Also, grain yield was increased to 13.27 and 12.90 t ha -1 in 2016 and 2017 seasons, respectively. Moreover, dimetindene maleate induces some of the indicators for salicylic acid and jasmonic acid pathways via gene expression. These genes include OsWRKY45, OsNPR1, AOS2, JAMYB and PBZ1 (OsPR10), recording 15.14-, 16.47-, 5.3-, 5.37- and 5.1-fold changes, respectively, 12-h postinoculation., Conclusion: The results overview investigated the effectiveness of dimetindene maleate for increasing rice resistance to blast disease through inducing SAR in rice plants under glasshouse and field conditions, which could be through the SA defense pathway by expression of genes (OsWRKY45 and OsNPR1). © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Key Message: This study focused on the role of CLE1-7 peptides as defense mediators, and showed that root-expressed CLE3 functions as a systemic signal to regulate defense-related gene expression in shoots. In the natural environment, plants employ diverse signaling molecules including peptides to defend themselves against various pathogen attacks. In this study, we investigated whether CLAVATA3/EMBRYO SURROUNDING REGION-RELATED (CLE) genes (CLE1-7) respond to biotic stimuli. CLE3 showed significant up-regulation upon treatment with flg22, Pep2, and salicylic acid (SA). Quantitative real-time PCR (qRT-PCR) analysis revealed that CLE3 expression is regulated by the NON-EXPRESSOR OF PR GENES1 (NPR1)-dependent SA signaling and flg22-FLAGELLIN-SENSITIVE 2 (FLS2) signaling pathways. We demonstrated that SA-induced up-regulation of CLE3 in roots was required for activation of WRKY33, a gene involved in the regulation of systemic acquired resistance (SAR), in shoots, suggesting that CLE3 functions as a root-derived signal that regulates the expression of defense-related genes in shoots. Microarray analysis of transgenic Arabidopsis lines overexpressing CLE3 under the control of a β-estradiol-inducible promoter revealed that root-confined CLE3 overexpression affected gene expression in both roots and shoots. Comparison of CLE2- and CLE3-induced genes indicated that CLE2 and CLE3 peptides target a few common but largely distinct downstream genes. These results suggest that root-derived CLE3 is involved in the regulation of systemic rather than local immune responses. Our study also sheds light on the potential role of CLE peptides in long-distance regulation of plant immunity., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Salicylic acid (SA) is a natural phenolic compound in plants with many beneficial effects for humans. The anxiolytic effect of this compound has been reported in animal models, but its mechanism of action remains unclear. In this study, by using the fear potentiated plus maze test, we evaluated the effect of salicylic acid on the gene expression of the main form of GABA (gamma aminobutyric acid) synthesizing enzyme i.e., the enzyme glutamic acid decarboxylase 67 (GAD67) which is called GAD1, in the ventral subiculum of the hippocampus, one of the main brain structures, in anxiety circuits. Also, the hypnotic effect of Salicylic acid was evaluated., Methods: Animals were divided into the solvent, (SA) and diazepam treated groups (n = 6). For evaluating the anxiolytic effect of Salicylic acid, animals were subjected to 2 h of isolation, before placing them in the elevated plus maze (EPM). Afterward, the ventral part of the hippocampus was removed for evaluating the change in GAD1 gene expression by the reverse transcription-quantitative polymerase chain reaction (RTqPCR) technique. The hypnotic effect of Salicylic acid was evaluated in the ketamine induced sleeping test., Results: Salicylic acid at 10 and 30 (mg/kg) increased time spent and entries to the open arms in the (EPM) (p < 0.05). (RTqPCR) revealed that 30 mg/kg of Salicylic acid increased GAD1 gene expression (p < 0.001). Salicylic acid (30 and 300 mg/kg) also increased the duration of sleep, in ketamine induced sleeping test (p < 0.05)., Conclusion: Our results showed that Salicylic acid has anxiolytic and hypnotic effects and it exerts its anxiolytic effect partly, via up the regulation of GAD1 in the ventral part of the hippocampus., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: Salicylic acid (SA) is an important regulator of genes involved in plant defense and pathogen-triggered systemic acquired resistance (SAR). Coconut is an important crop affected by several pathogens. Reported evidence suggests SA involvement in defense responses, including SAR in coconut., Objective: To identified differentially expressed genes in leaf and root tissues of coconut plantlets, as a result of SA, that might be involved in coconut defense responses., Methods: Comparative transcriptomic analysis by RNA-Seq of leaf and root tissues from in vitro coconut plantlets unexposed and exposed to SA 2.5 mM for 48 h. And in silico validation of gene expression by qRT-PCR., Results: We identified 4615 and 3940 differentially expressed unigenes (DEUs) in leaf and root tissues respectively. Our GO analysis showed functional categories related to the induction of defense responses, such as "systemic acquired resistance" and highly enriched hormone categories, such as abscisic acid. The most abundant KEGG pathway in our results was "Biosynthesis of antibiotics". Our findings support that exogenous application of SA to plantlets induced the activation of PRs, RGAs, ICS2, NLTP2, PER4, TRXM and some WRKYs mediated by NPR1-dependent pathways. Also, we found DEUs, such as BZR1, HSL1, and WHY2 that support that SA could regulate defense-related genes through NPR1-independent pathways., Conclusion: The present study of massive data analysis carried out on coconut plantlets exposed to SA, generates valuable information that increases our understanding of defense molecular mechanisms in coconut and open new venues for research for the improvement of management of coconut diseases., (© 2021. The Genetics Society of Korea.)

Rice plants contain high basal levels of salicylic acid (SA), but some of their functions remain elusive. To elucidate the importance of SA homeostasis in rice immunity, we characterized four rice SA hydroxylase genes ( OsSAHs ) and verified their roles in SA metabolism and disease resistance. Recombinant OsSAH proteins catalyzed SA in vitro, while OsSAH3 protein showed only SA 5-hydroxylase (SA5H) activity, which was remarkably higher than that of other OsSAHs that presented both SA3H and SA5H activities. Amino acid substitutions revealed that three amino acids in the binding pocket affected SAH enzyme activity and/or specificity. Knockout OsSAH 2 and OsSAH3 ( sah KO) genes conferred enhanced resistance to both hemibiotrophic and necrotrophic pathogens, whereas overexpression of each OsSAH gene increased susceptibility to the pathogens. sah KO mutants showed increased SA and jasmonate levels compared to those of the wild type and OsSAH -overexpressing plants. Analysis of the OsSAH3 promoter indicated that its induction was mainly restricted around Magnaporthe oryzae infection sites. Taken together, our findings indicate that SA plays a vital role in immune signaling. Moreover, fine-tuning SA homeostasis through suppression of SA metabolism is an effective approach in studying broad-spectrum disease resistance in rice.

Under salt stress conditions, plant growth is reduced due to osmotic, nutritional and oxidative imbalance. However, salicylic acid acts in the mitigation of this abiotic stress by promoting an increase in growth, photosynthesis, nitrogen metabolism, synthesis of osmoregulators and antioxidant enzymes. In this context, the objective was to evaluate the effect of salicylic acid doses on the growth and physiological changes of eggplant seedlings under salt stress. The experiment was conducted in a greenhouse, where the treatments were distributed in randomized blocks using a central composite matrix Box with five levels of electrical conductivity of irrigation water (CEw) (0.50; 1.08; 2.50; 3.92 and 4.50 dS m-1), associated with five doses of salicylic acid (SA) (0.00; 0.22; 0.75; 1.28 and 1.50 mM), with four repetitions and each plot composed of three plants. At 40 days after sowing, plant height, stem diameter, number of leaves, leaf area, electrolyte leakage, relative water content, and total dry mass were determined. ECw and SA application influenced the growth and physiological changes of eggplant seedlings. Increasing the ECw reduced growth in the absence of SA. Membrane damage with the use of SA remained stable up to 3.9 dS m-1 of ECw. The relative water content independent of the CEw increased with 1.0 mM of SA. The use of SA at the concentration of 1.0 mM mitigated the deleterious effect of salinity on seedling growth up to 2.50 dS m-1 of ECw.

The present study investigated the role of salicylic acid (SA) in regulating morpho-anatomical adaptive responses of a wheat plant to waterlogging. Our pharmacological study showed that treatment of waterlogged wheat plants with exogenous SA promotes the formation axile roots and surface adventitious roots that originate from basal stem nodes, but inhibits their elongation, leading to the formation of a shallow root system. The treatment also enhanced axile root formation in non-waterlogged plants but with only slight reductions in their length and branch root formation. Exogenous SA enhanced the formation of root aerenchyma, a key anatomical adaptive response of plants to waterlogging. Consistent with these results, waterlogging enhanced SA content in the root via expression of specific isochorismate synthase ( ICS ; ICS1 and ICS2 ) and phenylalanine ammonia lyase ( PAL ; PAL4 , PAL5 and PAL6 ) genes and in the stem nodes via expression of specific PAL ( PAL5 and PAL6 ) genes. Although not to the same level observed in waterlogged plants, exogenous SA also induced aerenchyma formation in non-waterlogged plants. The findings of this study furthermore indicated that inhibition of ethylene synthesis in SA treated non-waterlogged and waterlogged plants does not have any effect on SA-induced emergence of axile and/or surface adventitious roots but represses SA-mediated induction of aerenchyma formation. These results highlight that the role of SA in promoting the development of axile and surface adventitious roots in waterlogged wheat plants is ethylene independent while the induction of aerenchyma formation by SA requires the presence of ethylene.

DEAD-box proteins are a large family of RNA helicases that play important roles in almost all cellular RNA processes in model plants. However, little is known about this family of proteins in crops such as soybean. Here, we identified 80 DEAD-box family genes in the Glycine max (soybean) genome. These DEAD-box genes were distributed on 19 chromosomes, and some genes were clustered together. The majority of DEAD-box family proteins were highly conserved in Arabidopsis and soybean, but Glyma.08G231300 and Glyma.14G115100 were specific to soybean. The promoters of these DEAD-box genes share cis -acting elements involved in plant responses to MeJA, salicylic acid (SA), low temperature and biotic as well as abiotic stresses; interestingly, half of the genes contain nodulation-related cis elements in their promoters. Microarray data analysis revealed that the DEAD-box genes were differentially expressed in the root and nodule. Notably, 31 genes were induced by rhizobia and/or were highly expressed in the nodule. Real-time quantitative PCR analysis validated the expression patterns of some DEAD-box genes, and among them, Glyma.08G231300 and Glyma.14G115100 were induced by rhizobia in root hair. Thus, we provide a comprehensive view of the DEAD-box family genes in soybean and highlight the crucial role of these genes in symbiotic nodulation.

Temperature is a key factor influencing plant growth and productivity, but its sudden rise can cause severe consequences on crop performances. Early sowing and application of growth promoting agents as a foliar spray can be a sustainable approach to cope with high temperature stress at grain filling stage of cereal crops. Therefore, a test was designed to explore the potential of different growth helping agents including sorghum water extract (SWE, 10 ml L-1), moringa leaf extract (MLE, 3%), hydrogen peroxide (H2O2, 2 μM), salicylic acid (SA, 50 mg L-1) and ascorbic acid (ASA, 50 mg L-1) as foliar agents at different sowing dates (early and optimum) to cope with temperature stress in maize. The results stated that foliar application of growth promoting substances successfully persuaded high temperature tolerance at reproductive phase of maize in early and optimum sowings when compared to control. However, SWE + ASA, MLE + H2O2 and SWE + ASA + SA + H2O2 were the best combinations for improving growth, development, and physiological variables under both sowing dates even under suboptimal temperature. All foliar applications significantly increased maize grain and biological yields while maximum was observed in SWE + ASA followed by SWE + ASA + SA + H2O2 or MLE + H2O2 that were statistically at par with ASA + SA + H2O2 but plants without spray or distilled water application did not improve grain and biological yields. Overall, the foliar applications of growth promoting substances enable the plant to enhance its growth, development, morphology, yield and biochemical variables., Competing Interests: The authors have declared that no competing interests exist.

Frost injury is one of the major limiting factors to horticultural crops production and distribution. Despite numerous efforts and researches concerning freezing injury reduction, it still accounts for more than 50% of the fruit losses in the horticulture sector. In the present investigation, we aimed to investigate the effects of different antifreeze compounds (Tiofer®, Cropaid®, Bio-Bloom®, amino acid (mixture), salicylic acid, and water (control)) on pistachio trees behavior under low-temperature regimes (2 °C, 0 °C, -2 °C, -4 °C, -6 °C, and spring natural temperature). The applied chemicals improved the osmolyte content during the cold stress. Tiofer® and Cropaid® could increase the proline content better than other compounds. Salicylic acid and Cropaid® application increased the guaiacol peroxidase (GPX) content better than other compounds. For ascorbate peroxidase (APX), Tiofer® and Bio-Bloom®, and for catalase (CAT), Tiofer®, Cropaid®, and salicylic acid performed better. Applying chemicals also improved the photosynthetic pigments under cold stress. Among all treatments, Tiofer® and Bio-Bloom® improved the chlorophyll a (Chla), while chlorophyll b (Chlb) better improved by Tiofer® and Cropaid®; moreover, carotenoids had better increase in Cropaid®, amino acid, and salicylic acid treatments. All applied chemicals except Tiofer® had a good effect on the anthocyanin content increase under cold stress. In conclusion, based on the findings presented here, applying antifreeze compounds, such as Tiofer®, Cropaid® Bio-Bloom®, salicylic acid, and amino acid, could effectively ameliorate the adverse effects of cold stress. Osmolytes and antioxidant (GPX, APX, CAT) contents, photosynthetic pigments (chlorophyll a and b and carotenoid), and anthocyanins were improved. Among all applied antifreezes, Tiofer® and Cropaid® were the most effective ones., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Plant hormones regulate growth, development, and defense against biotic and abiotic stresses. Salicylic acid (SA), ethylene (ET), and jasmonate (JA) are major phytohormones that control the defense against pathogens. SA and JA primarily regulate resistance against biotrophic and necrotrophic pathogens, respectively. NPR1 is the key regulator of SA signaling in plants. AtOZF1 function has recently been ascribed to promote both NPR1- dependent and -independent SA signaling. However, the role of AtOZF1 in JA signaling was not known. Here we report AtOZF1 as a positive regulator of JA signaling in Arabidopsis. The atozf1 mutants are more susceptible to the necrotrophic pathogen Botrytis cinerea than wildtype (WT) plants. AtOZF1 positively regulates the expression of JA inducible genes like PDF1.2, VSP2, THI2.1, and ORA59 . AtOZF1 takes part in SA-JA cross-talk to an extent similar to that of NPR1. AtOZF1 is essential for the activation of PDF1.2 expression upon exogenous methyl-jasmonate (MeJA) application. Intriguingly, SA can significantly promote MeJA-induced PDF1.2 expression in the absence of AtOZF1. Altogether our results reveal a novel SA-JA interaction pathway in plants.

Although protein secretion was previously believed to be solely via ER/Golgi pathways, Golgi-independent secretion has now been described in both animals and plants. Secretion of the mannitol catabolic enzyme mannitol dehydrogenase (MTD) in response to the endogenous pathogen response signal salicylic acid (SA) was one of the first reports of unconventional protein secretion in plants. To begin assessing potential secretion-associated MTD protein interactors, we present here high-quality databases describing changes in MTD-interacting proteins following SA treatment of Arabidopsis thaliana cells expressing MTD., (© 2021 Wiley-VCH GmbH.)

Ligand inducible proteins that enable precise and reversible control of nuclear translocation of passenger proteins have broad applications ranging from genetic studies in mammals to therapeutics that target diseases such as cancer and diabetes. One of the drawbacks of the current translocation systems is that the ligands used to control nuclear localization are either toxic or prone to crosstalk with endogenous protein cascades within live animals. We sought to take advantage of salicylic acid (SA), a small molecule that has been extensively used in humans. In plants, SA functions as a hormone that can mediate immunity and is sensed by the nonexpressor of pathogenesis-related (NPR) proteins. Although it is well recognized that nuclear translocation of NPR1 is essential to promoting immunity in plants, the exact subdomain of Arabidopsis thaliana NPR1 (AtNPR1) essential for SA-mediated nuclear translocation is controversial. Here, we utilized the fluorescent protein mCherry as the reporter to investigate the ability of SA to induce nuclear translocation of the full-length NPR1 protein or its C-terminal transactivation (TAD) domain using HEK293 cells as a heterologous system. HEK293 cells lack accessory plant proteins including NPR3/NPR4 and are thus ideally suited for studying the impact of SA-induced changes in NPR1. Our results obtained using a stable expression system show that the TAD of AtNPR1 is sufficient to enable the reversible SA-mediated nuclear translocation of mCherry. Our studies advance a basic understanding of nuclear translocation mediated by the TAD of AtNPR1 and uncover a biotechnological tool for SA-mediated nuclear localization., (© 2021 Wiley Periodicals LLC.)

In recent years, numerous studies have shown that conversion of conventional drugs in ionic liquid (IL) formulation could be a successful strategy to improve their physicochemical properties or suggest a new route of administration. We report the synthesis and detailed characterization of eight salicylic acid-based ILs (SA-ILs) containing cation non-polar or aromatic amino acid esters. Using in vitro assays, we preliminary evaluated the therapeutic potency of the novel SA-ILs. We observed that conversion of the SA into ionic liquids led to a decrease in its cytotoxicity toward NIH/3T3 murine embryo fibroblasts and human HaCaT keratinocytes. It should be mentioned is that all amino acid alkyl ester salicylates [AAOR][SA] inhibit the production of the proinflammatory cytokine IL-6 in LPS-stimulated keratinocytes. Moreover, keratinocytes, pretreated with [PheOMe][SA] and [PheOPr][SA] seem to be protected from LPS-induced inflammation. Finally, the novel compounds exhibit a similar binding affinity to bovine serum albumin (BSA) as the parent SA, suggesting a similar pharmacokinetic profile. These preliminary results indicate that SA-ILs, especially those with [PheOMe], [PheOPr], and [ValOiPr] cation, have the potential to be further investigated as novel topical agents for chronic skin diseases such as psoriasis and acne vulgaris.

Haematococcus pluvialis accumulates a large amount of astaxanthin under various stresses, e.g., blue light and salicylic acid (SA). However, the metabolic response of H. pluvialis to blue light and SA is still unclear. We investigate the effects of blue light and SA on the metabolic response in H. pluvialis using both transcriptomic and proteomic sequencing analyses. The largest numbers of differentially expressed proteins (DEPs; 324) and differentially expressed genes (DEGs; 13,555) were identified on day 2 and day 7 of the treatment with blue light irradiation (150 μmol photons m -2 s -1 ), respectively. With the addition of SA (2.5 mg/L), a total of 63 DEPs and 11,638 DEGs were revealed on day 2 and day 7, respectively. We further analyzed the molecular response in five metabolic pathways related to astaxanthin synthesis, including the astaxanthin synthesis pathway, the fatty acid synthesis pathway, the heme synthesis pathway, the reactive oxygen species (ROS) clearance pathway, and the cell wall biosynthesis pathway. Results show that blue light causes a significant down-regulation of the expression of key genes involved in astaxanthin synthesis and significantly increases the expression of heme oxygenase, which shows decreased expression by the treatment with SA. Our study provides novel insights into the production of astaxanthin by H. pluvialis treated with blue light and SA.

Wheat is an important cereal crop, which is adversely affected by water deficit stress. The effect of induced stress can be reduced by the application of salicylic acid (SA). With the objective to combat drought stress in wheat, an experiment was conducted in greenhouse under hydroponic conditions. The treatments consisted of (a) no drought (DD0 = 0 MPa), mild drought (DD1 = -0.40 MPa) and severe drought (DD2 = -0.60 MPa) by applying PEG-8000, (b) two contrasting wheat varieties Barani-17 (drought tolerant) and Anaj-17 (drought-sensitive), and (c) foliar treatments of salicylic acid (0, 50 mM, 75 mM, and 100 mM). Evaluation of wheat plants regarding biochemical, physiological, and morphological attributes were rendered after harvesting of plants. Statistically, maximum shoot and root fresh and dry weights (18.77, 11.15 and 1.99, 1.81 g, respectively) were recorded in cultivar Barani-17 under no drought condition with the application of SA (100 mM). While, minimum shoot and root fresh and dry weights (6.65, 3.14 and 0.73, 0.61 g, respectively) were recorded in cultivar Anaj-2017 under mild drought stress without SA application. The maximum shoot length (68.0 cm) was observed in cultivar Barani-2017 under no drought condition with the application of SA (100 mM). While, maximum root length (59.67 cm) was recorded in cultivar Anaj-17 under moderate drought stress without application of SA. Further, minimum shoot length (28.67 cm) was recorded in Anaj-17 under moderate drought stress without SA application. Minimum root length (38.67 cm) was recorded in cultivar Barani-17 under no drought condition without SA application. Furthermore, maximum physio-biochemical traits, including membrane stability index (MSI), chlorophyl content, photosynthetic rates, stomatal conductance, antioxidant enzymatic activities and relative water content (RWC) were found highest in cultivar Barani-17 under no drought stress and SA application at 100 mM. However, minimum values of these traits were recorded in cultivar Anaj-17 under severe drought stress without SA application. Our results also demonstrated that under severe drought, application of SA at 100 mM significantly increased leaf nitrogen (N), phosphrus (P) and potassium (K) contents and cultivar Barani-17 demonstrated significantly higher values than Anaj-17. The obtained results also indicated that the cultivation of wheat under drought stress conditions noticeably declines the morphological, physiological, and biochemical attributes of the plants. However, the exogenous application of SA had a positive impact on wheat crop for enhancing its productivity., Competing Interests: The authors have declared that no competing interests exist.

Main Conclusion: Salicylic acid (SA) and drought stress promote more flowering in sweet orange. The physiological response and molecular mechanism underlying stress-induced floral initiation were discovered by transcriptome profiling. Numerous flowering-regulated genes were identified, and ectopically expressed CsLIP2A promotes early flowering in Arabidopsis. Floral initiation is a critical developmental mechanism associated with external factors, and citrus flowering is mainly regulated by drought stress. However, little is known about the intricate regulatory network involved in stress-induced flowering in citrus. To understand the molecular mechanism of floral initiation in citrus, flower induction was performed on potted Citrus sinensis trees under the combined treatment of salicylic acid (SA) and drought (DR). Physiological analysis revealed that SA treatment significantly normalized the drastic effect of drought stress by increasing antioxidant enzyme activities (SOD, POD, and CAT), relative leaf water content, total chlorophyll, and proline contents and promoting more flowering than drought treatment. Analysis of transcriptome changes in leaves from different treatments showed that 1135, 2728 and 957 differentially expressed genes (DEGs) were revealed in response to DR, SD (SA + DR), and SA (SA + well water) treatments in comparison with the well watered plants, respectively. A total of 2415, 2318 and 1933 DEGs were expressed in DR, SD, and SA in comparison with water recovery, respectively. Some key flowering genes were more highly expressed in SA-treated drought plants than in DR-treated plants. GO enrichment revealed that SA treatment enhances the regulation and growth of meristem activity under drought conditions, but no such a pathway was found to be highly enriched in the control. Furthermore, we focused on various hormones, sugars, starch metabolism, and biosynthesis-related genes. The KEGG analysis demonstrated that DEGs enriched in starch sucrose metabolism and hormonal signal transduction pathways probably account for stress-induced floral initiation in citrus. In addition, a citrus LIPOYLTRANSFERSAE 2A homologous (LIP2A) gene was upregulated by SD treatment. Ectopic expression of CsLIP2A exhibited early flowering in transgenic Arabidopsis. Taken together, this study provides new insight that contributes to citrus tree floral initiation under the SA-drought scenario as well as an excellent reference for stress-induced floral initiation in woody trees., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Wetland plants play a major role in the process of wastewater treatment in constructed wetlands (CWs). The inhibitory effect of salt stress on plants may reduce the performance of CWs. In this study, salicylic acid (SA) and/or calcium ion (Ca 2+ ) were used for root pretreatment to alleviate the salt stress in Iris pseudacorus L. The results showed that root pretreatment with SA and/or Ca 2+ improved the response of Iris pseudacorus L. to salinity by increasing growth, photosynthetic pigments, Pro content, enzymes activities and K + content. In addition, SA and/or Ca 2+ application in saline conditions decreased the relative conductivity and content of malondialdehyde. RNA-seq analysis showed the expression of hormone signaling genes, potassium ion transporter genes, oxidative stress genes and photosynthesis genes were up-regulated after pretreating with SA and CaCl 2 . In conclusion, the addition of SA and Ca 2+ could improve the saline wastewater treatment efficiency of CWs by enhancing the salt tolerance of Iris pseudacorus L., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Plants tailor immune responses to defend against pathogens with different lifestyles. In this process, antagonism between the immune hormones salicylic acid (SA) and jasmonic acid (JA) optimizes transcriptional signatures specifically to the attacker encountered. Antagonism is controlled by the transcription cofactor NPR1. The indispensable role of NPR1 in activating SA-responsive genes is well understood, but how it functions as a repressor of JA-responsive genes remains unclear. Here, we demonstrate that SA-induced NPR1 is recruited to JA-responsive promoter regions that are co-occupied by a JA-induced transcription complex consisting of the MYC2 activator and MED25 Mediator subunit. In the presence of SA, NPR1 physically associates with JA-induced MYC2 and inhibits transcriptional activation by disrupting its interaction with MED25. Importantly, NPR1-mediated inhibition of MYC2 is a major immune mechanism for suppressing pathogen virulence. Thus, NPR1 orchestrates the immune transcriptome not only by activating SA-responsive genes but also by acting as a corepressor of JA-responsive MYC2., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Drought has become a dangerous threat to reduce crop productivity throughout the world. Exogenous applications of regulators, micronutrients, and/or osmoprotectants for inducing drought-tolerance in field crops have been effectively adopted. A controlled pot study was performed to investigate the relative efficacy of salicylic acid (SA), zinc (Zn), and glycine betaine (GB) as foliar applications on the growth, tissues pigments content, relative water content (RWC), leaf gas-exchange, antioxidant enzymes activity, reactive oxygen species (ROS) accumulation, osmolytes contents, and the yield parameters of wheat plants subjected to two soil water conditions (85% field capacity: well-watered, 50% field capacity: water-deficient) during reproductive growth stages., Results: Water deficient conditions significantly decreased the growth, yield parameters, RWC, photosynthesis pigment, and gas-exchange attributes except for intercellular CO 2 concentration. However, foliar applications remarkably improved the growth and yield parameters under water deficit conditions. Under drought condition, exogenous applications of SA, Zn, and GB increased the grain yield pot - 1 by 27.99, 15.23 and 37.36%, respectively, as compared to the control treatment. Drought stress statistically increased the contents of hydrogen peroxide (H 2 O 2 ), superoxide anion radical (O 2 •- ), and malonaldehyde (MDA), and elevated the harmful oxidation to cell lipids in plants, however, they were considerably reduced by foliar applications. Foliar applications of SA, Zn, and GB decreased MDA content by 29.09, 16.64 and 26.51% under drought stress, respectively, as compared to the control treatment. Activities of all antioxidant enzymes, proline content, and soluble sugar were increased in response to foliar applications under water deficit conditions., Conclusions: Overall, foliar application of GB, SA, and Zn compounds improved the drought-tolerance in wheat by decreasing the ROS accumulation, promoting enzymatic antioxidants, and increasing osmolytes accumulation. Finally, GB treatment was most effective in thoroughly assessed parameters of wheat followed by SA and Zn applications to alleviate the adverse effects of drought stress., (© 2021. The Author(s).)

Plants have developed sophisticated strategies to coordinate growth and immunity, but our understanding of the underlying mechanism remains limited. In this study, we identified a novel molecular module that regulates plant growth and defense in both compatible and incompatible infections. This module consisted of BZR1, a key transcription factor in brassinosteroid (BR) signaling, and EDS1, an essential positive regulator of plant innate immunity. We found that EDS1 interacts with BZR1 and suppresses its transcriptional activities. Consistently, upregulation of EDS1 function by a virulent Pseudomonas syringae strain or salicylic acid treatment inhibited BZR1-regulated expression of BR-responsive genes and BR-promoted growth. Furthermore, we showed that the cytoplasmic fraction of BZR1 positively regulates effector-triggered immunity (ETI) controlled by the TIR-NB-LRR protein RPS4, which is attenuated by BZR1's nuclear translocation. Mechanistically, cytoplasmic BZR1 facilitated AvrRps4-triggered dissociation of EDS1 and RPS4 by binding to EDS1, thus leading to efficient activation of RPS4-controlled ETI. Notably, transgenic expression of a mutant BZR1 that accumulates exclusively in the cytoplasm improved pathogen resistance without compromising plant growth. Collectively, these results shed new light on plant growth-defense coordination and reveal a previously unknown function for the cytoplasmic fraction of BZR1. The BZR1-EDS1 module may be harnessed for the simultaneous improvement of crop productivity and pathogen resistance., (Copyright © 2021 The Author. Published by Elsevier Inc. All rights reserved.)

The priming of defence responses in pathogen-challenged model plants undergoes a preparation phase and an expression phase for defence function. However, the priming response in postharvest fruits has not been elucidated. Here, we found that 50 mM β-aminobutyric acid (BABA) treatment could induce two distinct pathways linked with TGA1-related systemic acquired resistance (SAR), resulting in the alleviation of Rhizopus rot in postharvest peach fruit. The first priming phase was elicited by BABA alone, leading to the enhanced transcription of redox-regulated genes and posttranslational modification of PpTGA1. The second phase was activated by an H 2 O 2 burst via up-regulation of PpRBOH genes and stimulation of the MAPK cascade on pathogen invasion, resulting in a robust defence. In the MAPK cascade, PpMAPKK5 was identified as a shortcut interacting protein of PpTGA1 and increased the DNA binding activity of PpTGA1 for the activation of salicylic acid (SA)-responsive PR genes. The overexpression of PpMAPKK5 in Arabidopsis caused the constitutive transcription of SA-dependent PR genes and as a result conferred resistance against the fungus Rhizopus stolonifer. Hence, we suggest that the BABA-induced priming defence in peaches is activated by redox homeostasis with an elicitor-induced reductive signalling and a pathogen-stimulated H 2 O 2 burst, which is accompanied by the possible phosphorylation of PpTGA1 by PpMAPKK5 for signal amplification., (© 2021 The Authors. Molecular Plant Pathology published by British Society for Plant Pathology and John Wiley & Sons Ltd.)

Colletotrichum is a plant pathogenic fungus which is able to infect virtually every economically important plant species. Up to now no common infection mechanism has been identified comparing different plant and Colletotrichum species. Plant hormones play a crucial role in plant-pathogen interactions regardless whether they are symbiotic or pathogenic. In this review we analyze the role of ethylene, abscisic acid, jasmonic acid, auxin and salicylic acid during Colletotrichum infections. Different Colletotrichum strains are capable of auxin production and this might contribute to virulence. In this review the role of different plant hormones in plant- Colletotrichum interactions will be discussed and thereby auxin biosynthetic pathways in Colletotrichum spp. will be proposed.

Objective: To investigate the effect of systemic administration of salicylate as a tinnitus inducing drug in the auditory cortex of guinea pigs., Methods: Extracellular recording of spikes of the primary auditory cortex and dorsocaudal areas in healthy male albino Hartley guinea pigs was continuously performed (pre- and post-salicylate)., Results: We recorded 160 single units in the primary auditory cortex from five guinea pigs and 156 single units in the dorsocaudal area from another five guinea pigs. The threshold was significantly elevated after the administration of salicylate in both the primary auditory cortex and dorsocaudal areas. The Q10dB value was significantly increased in the primary auditory cortex, whereas it has significantly decreased in the dorsocaudal area. Spontaneous firing activity was significantly decreased in the primary auditory cortex, whereas it has significantly increased in the dorsocaudal area., Conclusion: Salicylate induces significant changes in single units of both stimulated and spontaneous activity in the auditory cortex of guinea pigs. The spontaneous activity changed differently depending on its cortical areas, which may be due to the neural elements that generate tinnitus., Competing Interests: The authors have declared that no competing interests exist.

Abnormal melanogenesis and melanosome transport can cause skin pigmentation disorders that are often treated using ginseng-based formulation. We previously found that phenolic acid compounds in ginseng root could inhibit melanin production and as a skin-whitening agents. However, mechanisms of action underlying effects of ginseng phenolic acid monomers on melanogenesis remain unclear. This study was conducted to investigate effects of salicylic acid, a main ginseng root phenolic acid component, on melanogenesis and melanosome functions in melanocytes of zebrafish and other species. Salicylic acid exhibited no cytotoxicity and reduced melanin levels and tyrosinase activity in B16F10 murine melanoma cells and normal human epidermal melanocytes regardless of prior cell stimulation with α-melanocyte stimulating hormone. Additionally, salicylic acid treatment reduced expression of melanogenic enzymes tyrosinase, tyrosinase-related protein 1 and tyrosinase-related protein 2, while reducing expression of their master transcriptional regulator, microphthalmia-associated transcription factor. Moreover, reduced phosphorylation of cAMP response-element binding protein was observed due to reduced cAMP levels resulting from salicylic acid inhibition of upstream signal regulators (adenylyl cyclase and protein kinase A). Furthermore, salicylic acid treatment suppressed expression of transport complex-associated proteins melanophilin and myosin Va in two UVB-treated melanocytic cell lines, suppressed phagocytosis of fluorescent microspheres by UVB-stimulated human keratinocytes (HaCaT), inhibited protease-activated receptor 2 activation by reducing both Ca 2+ release and activation of phosphoinositide 3 kinase/AKT and mitogen-activated protein kinases and induced anti-melanogenic effects in zebrafish. Collectively, these results indicate that salicylic acid within ginseng root can inhibit melanocyte melanogenesis and melanin transport, while also suppressing keratinocyte phagocytic function., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Plant growth regulatory substances play a significant role in maintaining developmental and physiological processes in plants under abiotic stress. Apart from traditional plant hormones, the phenolic acids, salicylic acid (SA) and gallic acid (GaA), are emerging players with pivotal roles in alleviating various environmental perturbations. The present study compared the stress alleviation effect of these two phenolic acids in wheat (Triticum aestivum L. 'Gönen-98') seedling whose seeds were used in this study pre-treated with increasing doses of gamma irradiation (IR, 100 > 400 Gy). Leaves from seedlings hydroponically grown for 10 days in medium containing 100 μmol/l SA and GaA were used in the measurements and determinations. Accordingly, exogenous treatment with SA and GaA significantly improved plant growth and photosynthetic activity and regulated stress-induced osmolyte accumulation against γ-irradiation. Treatments also led to significant reductions in TBARS and H 2 O 2 contents. Antioxidant enzyme activities were further stimulated by SA and GaA treatment in comparison to IR alone. The phenolic pool including phenolic acids and GSH content in whole seedlings were promoted by IR and further SA and GaA applications. Contents in phenolic acids liberated from soluble free, soluble ester-conjugated and soluble glycoside-conjugated SA and GaA contents in roots and leaves increased following SA and GaA treatments alone in comparison to the control and IR groups. The present results indicate that SA and GaA can alleviate the ameliorative effects of IR, leading to further oxidative stress, and can improve the tolerance of stressed wheat seedlings by stimulating enzymatic and non-enzymatic antioxidant defence system components., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

Key Message: MiWRKY53 is expressed in response to various stresses and hormones. Although it is localized in the nucleus, it shows no transcriptional activation. Role of SA-mediated plant defence response is demonstrated. WRKY transcription factors are one the largest gene families in plants involved in almost every process in plants including development, physiological processes, and stress response. Salicylic acid (SA) is key regulator of biotic stress against various pathogens in plants acting via its multiple mechanisms to induce defence response. Herein, we have identified and functionally validated WRKY53 from mulberry (Morus indica var. K2). MiWRKY53 expressed differentially in response to different stress and hormonal treatments. MiWRKY53 belongs to group III of WKRY gene family, localized in nucleus, and lacks transcriptional activation activity in yeast. Hormone responsive behaviour of MiWRKY53 Arabidopsis overexpression (OE) transgenics preferentially was noted in root growth assay in response to Salicylic acid (SA). Arabidopsis overexpression plants also displayed alteration in leaf phenotype having wider leaves than the wild-type plants. PR-1 transcripts were higher in MiWRKY53 Arabidopsis OE plants and they displayed resistance towards biotrophic pathogen Pseudomonas syringae PstDC3000. MiWRKY53 Mulberry OE transgenics also depicted SA-responsive behaviour. Several hormones and stress-related cis-acting elements were also identified in the 1.2-Kb upstream regulatory region (URR) of MiWRKY53. Functional characterization of full-length promoter region revealed that it is induced by SA and further analysis of deletion constructs helped in the identification of minimal promoter responsible for its inducibility by SA. Altogether, the findings from this study point towards the SA preferential behaviour of MiWRKY53 and its function as regulator of plant defence response through SA-mediated mechanisms., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Carnivorous plants feed on animal prey, mainly insects, to get additional nutrients. This carnivorous syndrome is widely investigated and reported. In contrast, reports on herbivores feeding on carnivorous plants and related defenses of the plants under attack are rare. Here, we studied the interaction of a pitcher plant, Nepenthes x ventrata, with a generalist lepidopteran herbivore, Spodoptera littoralis, using a combination of LC/MS-based chemical analytics, choice and feeding assays. Chemical defenses in N. x ventrata leaves were analyzed upon S. littoralis feeding. A naphthoquinone, plumbagin, was identified in Nepenthes defense against herbivores and as the compound mainly responsible for the finding that S. littoralis larvae gained almost no weight when feeding on Nepenthes leaves. Plumbagin is constitutively present but further 3-fold increased upon long-term (> 1 day) feeding. Moreover, in parallel de novo induced trypsin protease inhibitor (TI) activity was identified. In contrast to TI activity, enhanced plumbagin levels were not phytohormone inducible, not even by defense-related jasmonates although upon herbivory their level increased more than 50-fold in the case of the bioactive jasmonic acid-isoleucine. We conclude that Nepenthes is efficiently protected against insect herbivores by naphthoquinones acting as phytoanticipins, which is supported by additional inducible defenses. The regulation of these defenses remains to be investigated., Competing Interests: The authors have declared that no competing interests exist.

Hormones orchestrate the physiology of organisms. Measuring the activity of defense hormone-responsive genes can help understanding immune signaling and facilitate breeding for plant health. However, different from model species like Arabidopsis, genes that respond to defense hormones salicylic acid (SA) and jasmonic acid (JA) have not been disclosed in the soybean crop. We performed global transcriptome analyses to fill this knowledge gap. Upon exogenous application, endogenous levels of SA and JA increased in leaves. SA predominantly activated genes linked to systemic acquired resistance and defense signaling whereas JA mainly activated wound response-associated genes. In general, SA-responsive genes were activated earlier than those responding to JA. Consistent with the paradigm of biotrophic pathogens predominantly activating SA responses, free SA and here identified most robust SA marker genes GmNIMIN1, GmNIMIN1.2 and GmWRK40 were induced upon inoculation with Phakopsora pachyrhizi, whereas JA marker genes did not respond to infection with the biotrophic fungus. Spodoptera exigua larvae caused a strong accumulation of JA-Ile and JA-specific mRNA transcripts of GmBPI1, GmKTI1 and GmAAT whereas neither free SA nor SA-marker gene transcripts accumulated upon insect feeding. Our study provides molecular tools for monitoring the dynamic accumulation of SA and JA, e.g. in a given stress condition., (© 2021. The Author(s).)

Salicylic acid (SA), a phytohormone, has been considered to be a key regulator mediating plant defence against pathogens. It is still vague how SA activates plant defence against herbivores such as chewing and sucking pests. Here, we used an aphid-susceptible wheat variety to investigate Sitobion avenae response to SA-induced wheat plants, and the effects of exogenous SA on some defence enzymes and phenolics in the plant immune system. In SA-treated wheat seedlings, intrinsic rate of natural increase (rm), fecundity and apterous rate of S. avenae were 0.25, 31.4 nymphs/female and 64.4%, respectively, and significantly lower than that in the controls (P < 0.05). Moreover, the increased activities of phenylalanine-ammonia-lyase, polyphenol oxidase (PPO) and peroxidase in the SA-induced seedlings obviously depended on the sampling time, whereas activities of catalase and 4-coumarate:CoA ligase were suppressed significantly at 24, 48 and 72 h in comparison with the control. Dynamic levels of p-coumaric acid at 96 h, caffeic acid at 24 and 72 h and chlorogenic acid at 24, 48 and 96 h in wheat plants were significantly upregulated by exogenous SA application. Nevertheless, only caffeic acid content was positively correlated with PPO activity in SA-treated wheat seedlings (P = 0.031). These findings indicate that exogenous SA significantly enhanced the defence of aphid-susceptible wheat variety against aphids by regulating the plant immune system, and may prove a potential application of SA in aphid control.

The current study investigated the putative role of salicylic acid (SA) in modulating Pb 2+ -induced DNA and oxidative damage in Allium cepa roots. Pb 2+ exposure enhanced free radical generation and reduced DNA integrity and antioxidant machinery after 24 h; however, SA pre-treatment (for 24 h) ameliorated Pb 2+ toxicity. Pb 2+ exposure led to an increase in malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ) accumulation and enhanced superoxide radical and hydroxyl radical levels. SA improved the efficiency of enzymatic antioxidants (ascorbate and guaiacol peroxidases [APX, GPX], superoxide dismutases [SOD], and catalases [CAT]) at 50-μM Pb 2+  concentration. However, SA pre-treatment could not improve the efficiency of CAT and APX at 500 μM of Pb 2+ treatment. Elevated levels of ascorbate and glutathione were observed in A. cepa roots pre-treated with SA and exposed to 50 μM Pb 2+ treatment, except for oxidized glutathione. Nuclear membrane integrity test demonstrated the ameliorating effect of SA by reducing the number of dark blue-stained nuclei as compared to Pb 2+ alone treatments. SA was successful in reducing DNA damage in cell exposed to higher concentration of Pb 2+ (500 μM) as observed through comet assay. The study concludes that SA played a major role in enhancing defense mechanism and protecting against DNA damage by acclimatizing the plant to Pb 2+ -induced toxicity., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Aim: To investigate the anti-cancer potential of salicylic acid and cisplatin combination in HeLa cells and the underlying mechanism., Main Methods: Drugs and disease targets were extracted from DrugBank, BATMAN-TCM, STITCH, PharmMapper and Comparative Toxigenomics Database. Cytoscape 3.8.2 was used to merge the protein-protein interaction networks and select core targets. GO and KEGG analysis was done using Metascape and WebGestalt. Effect of salicylic acid and cisplatin alone and in combination on cells viability was studied by MTT assay. The type of interaction between salicylic acid and cisplatin was determined by CompuSyn. Apoptosis was evaluated by molecular docking, Rhodamine-123, DAPI, AO/EtBr staining, flow cytometry, qRT-PCR and western blotting. Metastasis was studied using scratch assay and western blotting. UHRF1 transient silencing was performed by siRNA., Key Findings: Out of 420, 1863 and 1362 respective targets of salicylic acid, cisplatin and cervical cancer, 18 core proteins were enriched in apoptosis and cell migration related pathways. IC50 value of cisplatin was reduced by 14 fold in combination with salicylic acid at IC20 (4 μM). There was loss of mitochondrial membrane potential and downregulation of UHRF1, pAkt, full length PARP and pro-caspase 3 expression. Transient silencing of UHRF1 also induced mitochondrial depolarization and apoptosis. The combination also exhibited anti-metastasis effect as it suppressed migration, upregulated PAX1 and downregulated MMP-2., Significance: Reduction in cisplatin concentration, enhanced anti-cancer effects and UHRF1 downregulation due to synergistic interaction between salicylic acid and cisplatin underscores the therapeutic importance of the combination to overcome chemo-resistance and side effects of cisplatin., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Peach fruit stored in the cold are susceptible to chilling injury. A pre-storage treatment with the natural hormone salicylic acid can alleviate chilling damage, although the mechanism is unclear. We found that a treatment with 1 μmol L -1 salicylic acid for 15 min prior to storage at 4 °C delayed and reduced fruit internal browning, a symptom of chilling injury. Salicylic acid had a large effect on sugar metabolism, increasing total soluble sugars via a substantial increase in sucrose content. The transcript abundance of genes related to sucrose biosynthesis and degradation was significantly regulated by salicylic acid, consistent with the changes in sucrose content. Salicylic acid treatment also increased the expression of two DREB cold stress-related proteins, transcriptional activators that regulate cold resistance pathways. The results show that salicylic acid alleviates chilling injury in peach by multiple mechanisms, including an increased content of sucrose and activation of cold response genes., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Mineral oils have been widely used in the pest control of several crops. However, their mode of action is poorly understood, especially in the case of their antifungal properties. The possible direct fungicidal activity and the stress-inducing capability of paraffin oil on grapevine were examined using Vitis vinifera L. cv 'Kékfrankos' cuttings and the fungus Erysiphe necator, the causal agent of powdery mildew., Results: Our experiments demonstrated that paraffin oil does not have fungicide activity on E. necator, but induces significant stress-related changes in grapevine physiology. Elevated H 2 O 2 production and the accumulation of the phytohormone salicylic acid were measured. Secondary thickening of the cell wall by lignin deposition and the accumulation of phenolic compounds were also observed. Some enzyme activities related to the detoxification of reactive oxygen species, disease response, and the synthesis of lignin were changed in accordance with the determined changes in cell wall composition and H 2 O 2 levels., Conclusion: The results suggest that paraffin oil induces stress responses on grapevine leaves through oxidative burst, and this response is systemized by salicylic acid. The accumulation of lignin and water-soluble phenolics may be directly responsible for the paraffin oil-induced resistance of grapevine against powdery mildew. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

The impact of salicylic acid (SA) on ethylene (ET) production and polyamine (PA) metabolism was investigated in wild type (WT) and ET receptor mutant Never ripe (Nr) tomato leaves under normal photoperiod and prolonged darkness. Nr displayed higher ET emanation compared to WT under control conditions and after SA treatments, but the ET signalling was blocked in these tissues. The accumulation of PAs was induced by 1 mM but not by 0.1 mM SA and was higher in WT than in Nr leaves. Upon 1 mM SA treatment, which caused hypersensitive response, illuminated leaves of WT showed high spermine (Spm) content in parallel with an increased expression of S-adenosylmethionine decarboxylase and Spm synthase (SlSPMS) suggesting that this process depended on the light. In Nr, however, Spm content and the expression of the SlSPMS gene were very low independently of the light conditions and SA treatments. This suggests that Spm synthesis needs functional ET perception. In WT leaves 1 mM SA enhanced putrescine (Put) synthesis by increasing the expression of Put biosynthesis genes, arginine and ornithine decarboxylases under darkness, while they were down-regulated in Nr. The activities of diamine (DAO) and polyamine oxidases (PAO), however, were generally higher in Nr compared to the WT after SA treatments. In Nr both SA applications increased the expression of SlPAO1 under normal photoperiod, while SlPAO2 was down-regulated in the dark suggesting a diverse role of PAOs in PA catabolism. These results indicated that ET could modulate the SA-induced PA metabolism in light-dependent manner., (Copyright © 2021. Published by Elsevier Masson SAS.)

In this research, eight local mung bean (Vigna radiata) varieties were analyzed for their performance against two levels of CdCl2 solution (0.3 and 0.5 mM) alone and priming with gibberellic acid (GA3) (100 μM), salicylic acid (SA) (50 μM) and proline (5 mM) solution prior to Cd exposure. Mung bean seedlings were analyzed for disturbance in cytological, morphological, biochemical and enzymatic parameters under cadmium stress. For cytological studies, 48 h grown mung bean seedlings root tips were used to prepare slides and studied for percent mitotic index (MI%) and to calculate percent C-mitosis, laggard, sticky and fragmented chromosomes, pictures were captured by a Nikon camera (DS-Fi 1 Japan) attached with a microscope. One-week grown mung seedlings were studied for growth traits, malondialdehyde (MDA), protein, proline and antioxidant enzymes. ANOVA and DMR test of this research revealed that all the tested mung bean varieties and treatments were significantly different regarding mitotic index and number of chromosomal aberrations. Both the Cd treatments exhibited increased total chromosomal aberrations with different types and a maximum decrease in MI%. In pretreated samples, GA3, SA and proline serve as mitigating agents that reduce mutagenic effects of Cd in mung bean by increasing MI% and decreasing chromosomal aberrations as compared to non-pretreated samples. Both the Cd treatments showed a decrease in all growth traits. Total proteins were also found to be significantly reduced in a dose-dependent manner in all genotypes. Cd treatment increased the activities of all antioxidant enzymes tested. Cd caused oxidative damage as indicated by elevated levels of MDA content in treated samples in comparison to control. Proline content levels were also high in Cd treated seedlings indicating stress. Results demonstrated that pretreatment with phytohormones and proline before Cd were found to improve all morphological parameters, by altering antioxidant enzymes activities along with a decrease in MDA and proline contents as well. It was further noticed that the performance of GA3 was better at 0.3 mM Cd treatment while SA was found to be a good mitigating agent at 0.5 mM Cd stress in all tested mung bean varieties. This research concluded less deleterious effects of Cd on AZRI-2006 while more sensitivity to NM-51 towards Cd. Priming with phytohormones and proline is a user-friendly, economical, and simple mitigation strategy to reduce Cd toxicity in plants and get better yield from contaminated lands., Competing Interests: The authors have declared that no competing interests exist.

We report the design and synthesis of a series of new 5-chloropyridinyl esters of salicylic acid, ibuprofen, indomethacin, and related aromatic carboxylic acids for evaluation against SARS-CoV-2 3CL protease enzyme. These ester derivatives were synthesized using EDC in the presence of DMAP to provide various esters in good to excellent yields. Compounds are stable and purified by silica gel chromatography and characterized using 1 H-NMR, 13 C-NMR, and mass spectral analysis. These synthetic derivatives were evaluated in our in vitro SARS-CoV-2 3CLpro inhibition assay using authentic SARS-CoV-2 3CLpro enzyme. Compounds were also evaluated in our in vitro antiviral assay using quantitative VeroE 6 cell-based assay with RNAqPCR. A number of compounds exhibited potent SARS-CoV-2 3CLpro inhibitory activity and antiviral activity. Compound 9a was the most potent inhibitor, with an enzyme IC 50 value of 160 nM. Compound 13b exhibited an enzyme IC 50 value of 4.9 µM. However, it exhibited a potent antiviral EC 50 value of 24 µM in VeroE6 cells. Remdesivir, an RdRp inhibitor, exhibited an antiviral EC 50 value of 2.4 µM in the same assay. We assessed the mode of inhibition using mass spectral analysis which suggested the formation of a covalent bond with the enzyme. To obtain molecular insight, we have created a model of compound 9a bound to SARS-CoV-2 3CLpro in the active site.

Background: In plants, basic leucine zipper transcription factors (TFs) play important roles in multiple biological processes such as anthesis, fruit growth & development and stress responses. However, systematic investigation and characterization of bZIP-TFs remain unclear in Chinese white pear. Chinese white pear is a fruit crop that has important nutritional and medicinal values., Results: In this study, 62 bZIP genes were comprehensively identified from Chinese Pear, and 54 genes were distributed among 17 chromosomes. Frequent whole-genome duplication (WGD) and dispersed duplication (DSD) were the major driving forces underlying the bZIP gene family in Chinese white pear. bZIP-TFs are classified into 13 subfamilies according to the phylogenetic tree. Subsequently, purifying selection plays an important role in the evolution process of PbbZIPs. Synteny analysis of bZIP genes revealed that 196 orthologous gene pairs were identified between Pyrus bretschneideri, Fragaria vesca, Prunus mume, and Prunus persica. Moreover, cis-elements that respond to various stresses and hormones were found on the promoter regions of PbbZIP, which were induced by stimuli. Gene structure (intron/exon) and different compositions of motifs revealed that functional divergence among subfamilies. Expression pattern of PbbZIP genes differential expressed under hormonal treatment abscisic acid, salicylic acid, and methyl jasmonate  in pear fruits by real-time qRT-PCR., Conclusions: Collectively, a systematic analysis of gene structure, motif composition, subcellular localization, synteny analysis, and calculation of synonymous (Ks) and non-synonymous (Ka) was performed in Chinese white pear. Sixty-two bZIP-TFs in Chinese pear were identified, and their expression profiles were comprehensively analyzed under ABA, SA, and MeJa hormones, which respond to multiple abiotic stresses and fruit growth and development. PbbZIP gene occurred through Whole-genome duplication and dispersed duplication events. These results provide a basic framework for further elucidating the biological function characterizations under multiple developmental stages and abiotic stress responses., (© 2021. The Author(s).)

The contribution of hydrogen sulphide (H 2 S) to salicylic acid (SA) induced lead (Pb) stress tolerance modulated by the ascorbate-glutathione (AsA-GSH) cycle was examined in pepper (Capsicum annuum L.) plants. One week after germination, pepper seedlings were sprayed with 0.5 mM SA once a day for a week. Thereafter, seedlings were grown under control (no Pb) or Pb stress (Pb-S treatment consisting of 0.1 mM PbCl 2 ) for a further 2 weeks. Lead stress reduced plant growth and leaf water status as well as the activities of dehydroascorbate reductase and monodehydroascorbate reductase. However, lead stress elevated leaf Pb, the proline contents, oxidative stress, activities of glutathione reductase and ascorbate peroxidase, as well as the endogenous H 2 S content. Supplements of SA resulted in improvements in growth parameters, biomass, leaf water status and AsA-GSH cycle-related enzyme activities, as well as increasing the H 2 S content. The positive effect of SA was further enhanced when sodium hydrosulphide was added. However, 0.1 mM hypotaurine (HT) treatment reversed the beneficial effect of SA by reducing the plant H 2 S content. Application of NaHS in combination with SA + HT suppressed the adverse effect of HT mainly by restoring the plant H 2 S content, suggesting that higher H 2 S content, induced by exogenous SA supply, resulted in elevated regulation of the AsA-GSH cycle., (© 2020 Scandinavian Plant Physiology Society.)

We have constructed two intra-molecularly shuffled promoters, namely S100 and D100. The S100 recombinant promoter (621 bp) was generated by ligation of 250 bp long upstream activation sequence (UAS) of Strawberry vein banding virus (SV10UAS; - 352 to - 102 relative to TSS) with its 371 bp long TATA containing core promoter domain (SV10CP; - 352 to + 19). Likewise, 726 bp long D100 promoter was constructed by fusion of 170 bp long UAS of Dahlia mosaic virus (DaMV14UAS; - 203 to - 33) with its 556 bp long core promoter domain (DaMV4CP; - 474 to + 82). S100 and D100 promoters showed 1.8 and 2.2 times stronger activities than that of the CaMV35S promoter. The activity of the promoters is comparable to that of the CaMV35S 2 promoter. Transcript analysis employing qRT-PCR and histochemical assays supported the above findings. Abscisic acid and salicylic acid induce the activity of the D100 promoter. Leaf protein obtained from Nicotiana tabacum plant expressing NSD2 gene (Nigella sativa L. defensin 2) driven by the D100 promoter showed antifungal activity against Alternaria alternata and Phoma exigua var. exigua and antibacterial activity against Pseudomonas aeruginosa and Staphylococcus aureus. Strong S100 and D100 promoters have potential to become efficient candidates for plant metabolic engineering and molecular pharming., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Chromium (Cr) phytotoxicity affirmed the need of mitigation strategies to remediate polluted soils and restricts its accumulation in the food chains. Salicylic acid (SA) and silicon (Si) play pivotal roles in stimulating the plant performance and stress resilience. So far, their interactive effects against Cr-phytotoxicities are less known. Thus, we evaluated the beneficial roles of alone or/and combine applications of SA and Si in mitigating the toxic effects of Cr in the leaves and roots of rice (Oryza sativa) seedlings. Results indicated that SA (10 μM) and/or Si (5 μM) markedly retrieved the Cr (100 μM) induced toxicities by minimizing the Cr-accretion in both leaves and roots, enhancing the performance of light harvesting pigments (total chlorophylls and carotenoids), water retention and accumulation of osmolytes (water-soluble protein and total soluble sugars) and ultimately improved the growth and biomass. Additionally, SA and/or Si maintained the ionic balance by enhancing the nutrients transport, upregulated the ascorbate-glutathione (AsA-GSH) cycle enzymes, minimized the extra accumulation of reactive oxygen species (ROS) (H 2 O 2 and O 2 •‒ ), malondialdehyde (MDA), recovered the membrane stability and damages in cellular ultrastructure in Cr-stressed rice plants. Overall findings suggested that SA underpins Si in mitigating the Cr-induced phytotoxicities on the above-reported parameters and combined applications of SA and Si were more effective than alone treatments. The uptake or cellular accumulation of Cr, osmoprotectants level and antioxidant defense system against oxidative stress can be considered as key toxicity biomarkers for the safe cultivation of rice in Cr-contaminated soils., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)